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Abstract Book Postersession Tuesday web-JNPC PDF
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Abstract book
• Plenary lectures
• Keynote lectures
• Short lectures
• Young Researchers Workshop
• Pre-conference Symposium
• Poster session Monday
• Poster session Tuesday
• Poster session Wednesday
Sponsors
The organizers acknowledge the support of the below sponsors and exhibitors - without
whom many of the activities would not have been feasible
Gold sponsor:
Silver sponsors:
Sponsors and exhibitors:
PRESENTATION OF POSTERS
Numbered poster boards for posters size A0 (portrait, 84.1 x 118.9 cm or 33.1 x 44.8 inches) and pins are
available. Posters are ordered according to the topics of JNPC2016, and you should find your poster number
in the electronic poster abstract book.
Poster sessions
Posters should be presented by the corresponding author during poster sessions Monday-Wednesday
13.00-14.45 according to the below schedule. Posters not presented by the authors will not be published in
the final abstract book in Planta Medica.
Monday Poster 1-365
Tuesday Poster 366-739
Wednesday Poster 750-1105 and YRW poster 1-12
Themes
Posters are sorted according to themes as shown below.
Theme 1: Advances in natural products research spurred by new technological developments
Theme 1.1: Poster 1-20 Theme 1.3: Poster 64-80
Theme 1.2: Poster 21-63 Theme 1.4: Poster 81-133
Theme 2: Sources of bioactive compounds
Theme 2.1: Poster 134-544 Theme 2.4: Poster 660-694
Theme 2.2: Poster 545-610 Theme 2.5: Poster 695-739
Theme 2.3: Poster 611-659
Theme 3: Sustainable sources of bioactive natural products
Theme 3.1: Poster 750-781 Theme 3.3: Poster 788-797
Theme 3.2: Poster 782-787
Theme 4: Natural products as preventive and curative medicine for humans and animals
Theme 4.1: Poster 798-840 Theme 4.3: Poster 931-991
Theme 4.2: Poster 841-930 Theme 4.4: Poster 992-1016
Theme 5: Production and regulatory aspects of herbal preparations
Theme 5.1: Poster 1017-1043 Theme 5.3: Poster 1086-1091
Theme 5.2: Poster 1044-1085
Late-added abstracts from various themes
Poster 1092-1105
POSTERSESSION TUESDAY
No
Title
and
authors
P366
Chemical
characterization
and
in
vitro
antibacterial
activity
of
Kalanchoe
brasiliensis
Cambess
(Crassulaceae)
Oscar
A.
S.
Mayorga,
Jônatas
R.
Florencio,
Jordana
D.
G.
de
Santana,
Carolina
Feres-‐Netto,
Dionnata
M.
Pedrosa,
Ygor
F.
G.
Costa,
Nícolas
C.
C.
Pinto,
Elita
Scio,
Orlando
V.
Sousa,
Maria
Silvana
Alves
P367
In
vitro
antibacterial
activity
of
Vernonia
polyanthes
Less.
leaf
rinse
extract
(Asteraceae):
pro-‐
specting
new
therapeutic
options
against
Staphylococcus
aureus
infections!
Jordana
D.
G.
de
Santana,
Jonatas
R.
Florêncio,
Luísa
M.
S.
de
Almeida,
Laura
S.
Fernandes,
Ademar
A.
da
Silva
Filho,
Marcos
José
Salvador,
Orlando
V.
Sousa,
Maria
Silvana
Alves
P368
Steroidal
saponins
from
Chlorophytum
deistelianum
Turibio
Tabopda,
Anne-‐Claire
Mitaine-‐Offer,
Thomas
Paululat,
Stéphanie
Delemasure,
Patrick
Dutartre,
Bonaventure
Tchaleu
Ngadjui,
Marie-‐Aleth
Lacaille-‐Dubois
P369
Triterpene
glycosides
from
plants
for
antibody
recognition
Elisa
Peroni,
Feliciana
Real
Fernández,
Caterina
Gheri,
Francesca
Nuti,
Anne-‐Claire
Mitaine-‐Offer,
Fran-‐
cesco
Lolli,
Marie-‐Aleth
Lacaille-‐Dubois,
Anna-‐Maria
Papini
P370
Chemical
characterisation
and
cytotoxicity
evaluation
of
Convolvulus
pluricaulis
Sieb.
ex
Spreng.
(Convolvulaceae)
extracts
towards
sensitive
and
multidrug-‐resistant
cancer
cells
Massimo
Tacchini,
Guglielmo
Paganetto,
Thomas
Efferth,
Gianni
Sacchetti,
Alessandra
Guerrini
P371
Survey
of
folk
use
and
gathering
of
plants
in
Karst
and
Gorjanci,
Slovenia
Mateja
Lumpert,
Samo
Kreft
P372
Anti-‐inflammatory
effects
and
toxicity
of
polysaccharide
fraction
from
Khaya
grandifoliola
stem
bark
Mediesse
Kengne
Francine,
Hasitha
Anantharaju,
Gangadhar
Matharasala,
Mbacham
F.
Wilfried,
Bou-‐
djeko
Thaddée,
Yogeeswari
Perumal
P373
Chemical
constituents
from
Agrimonia
pilosa
with
their
protein
tyrosine
phosphatase
and
ace-‐
tylcholinesterase
inhibitory
activities
Duc
H.
Nguyen,
Duc
D.
Le,
U
M.
Seo,
Thi
T.
Nguyen,
Jae
Soo
Choi,
Mi
H.
Woo
P374
Simultaneous
quantitation
of
five
bioactive
components
of
Menthae
Herba
by
HPLC/PDA
Mi
Hee
Woo,
Bing
Tian
Zhao,
Min
Je
Choi,
Young
Ho
Kim,
Jong
Seong
Kang
P375
Pharmacognostical
studies
on
‘Mo-‐Si-‐Pool’
Marjahan
Acter
Kazi,
Hye-‐Jin
Kim,
Woo
Sung
Park,
Khan
Khalil
Atif
Ali,
Mi-‐Jeong
Ahn
P376
A
prototype
traditional
Chinese
medicinal
plant
library
at
the
National
Cancer
Institute
Min
He,
Tanja
Grkovic,
Christopher
C.
Thornburg,
James
Whitt,
Rhone
Akee,
Jerell
Thompson,
John
Britt,
Libin
Jia,
Jeffrey
White,
David
Newman,
Barry
O’Keefe
P377
Evaluation
of
Rumex
crispus
extracts
as
novel
matrix
metalloproteinase-‐1
(MMP-‐1)
inhibitors
Mine
Uzun,
L.
Omur
Demirezer
P378
α-‐Glucosidase
inhibitory
prenylated
anthranols
from
Harungana
madagascariensis
Oluwatosin
O.
Johnson,
Ming
Zhao,
Jordan
Gunn,
Bernard
D.
Santarsiero,
Zhi-‐Qi
Yin,
Gloria
A.
Ayoola,
Herbert
A.
B.
Coker,
Chun-‐Tao
Che
P379
Agathisflavone
reduces
inflammatory
modulators
in
microglia
BV2
cells
Mireia
Boluda
Navarro,
Uchechukwu
Okorji,
Ravikanth
Velagapudi,
Priya
Jain,
Mutalib
Aderogba,
Olu-‐
mayokun
A
Olajide
P380
A
new
sesquiterpene
from
Lebanese
wild
carrot
inhibits
proliferation
of
human
acute
myeloid
leukemia
cells
Joelle
Boulos,
Ralph
Abi-‐Habib,
Mirvat
El-‐Sibai,
Wassim
Shebaby,
Costantine
F.
Daher,
Robin
I.
Taleb,
Mohamad
A.
Mroueh
P381
Marantodes
pumilum
L.
plant
extracts
induce
apoptosis,
cell
cycle
arrest
and
inhibit
cell
migra-‐
tion
and
invasion
on
prostate
cancer
cell
lines.
Mohd
Mukrish
Mohd
Hanafi,
Harisun
Yaakob,
Mohamad
Roji
Sarmidi,
Ramlan
Aziz,
Jose
Maria
Prieto
P382
HPLC-‐DAD-‐MS
analysis
of
extracts
from
flowers,
leaves,
fruits
and
branches
of
Ligustrum
vul-‐
gare
and
their
effect
on
cytokines
secretion
by
human
neutrophils
Monika
E.
Czerwińska,
Małgorzata
Warowny,
Anna
K.
Kiss
P383
Chemical
composition
and
antimicrobial
activity
of
the
essential
oil
of
the
leaves
of
Cupressus
macrocarpa
Amal
M.
Saad,
Magdy
M.
D.
Mohammed,
Mosad
A.
Ghareeb,
Wafaa
S.
Ahmed,
Mohamed
A.
Farid
P384
The
Chemistry
of
African
Croton
species
Moses
K
Langat,
Neil
Crouch,
Beth
Ndunda,
Jacob
O.
Midiwo,
Areej
Aldhaher,
Alaa
Alqahtani,
Dulcie
A
Mulholland
P385
Phytochemical
and
biological
investigation
of
Calliandra
surinamensis
as
a
potential
treatment
for
diabetes
Abdullah
Alzahrani,
Grainne
Abbott,
Louise
C.
Young,
John
Igoli,
Alexander
I.
Gray,
Valerie
A
Ferro
P386
The
effect
of
a
South
African
Helichrysum
sp.
against
important
pathogenic
mechanisms
of
Pro-‐
pionibacterium
acnes
M.
N.
de
Canha,
N.
Kishore,
N.
Lall
P387
α-‐amylase
and
α-‐glucosidase
inhibitor
activities
of
secondary
metabolites
from
Arcytophyllum
thymifolium
M.
De
Leo,
A.
Braca,
M.
B.
Vera
Saltos,
I.
Faraone,
N.
Malafronte,
L.
Milella,
N.
De
Tommasi
P388
Chrysophanol-‐
and
nepodin-‐8-‐O-‐β-‐D-‐glucopyranoside
from
Rumex
acetosella,
the
cytotoxicity
towards
drug
sensitive
and
multi-‐
drug
resistant
T
leukaemia
cancer
cells
Nadire
Ozenver,
Mohamed
Saeed,
Zuhal
Guvenalp,
L.
Omur
Demirezer,
Thomas
Efferth
P389
Cytotoxicity
of
main
anthraquinone
aglycons
towards
drug
sensitive
and
multi
drug
resistant
T
leukaemia
cancer
cells
Nadire
Ozenver,
Mohamed
Saeed,
L.
Omur
Demirezer,
Thomas
Efferth
P390
Comparative
antimicrobial
and
antioxidant
studies
of
two
closely
related
species
of
Saba
Nana
Ama
Mireku-‐Gyimah,
Kofi
Annan,
Ji-‐Kai
Liu,
Kwame
Sarpong
P391
Study
of
Pterocarpus
erinaceus,
a
promising
plant
from
Togo
to
treat
infectious
diseases
Nassifatou
Koko
Tittikpina,
Frédéric
Nana,
Stéphane
Fontanay,
Komlan
Batawila,
Koffi
Akpagana,
Gil-‐
bert
Kirsch,
Raphaël
E.
Duval,
Patrick
Chaimbault,
Claus
Jacob
P392
Cholinesterase
inhibitory
potentials
of
some
Turkish
medicinal
plants
Nehir
Unver
Somer,
Buket
Bozkurt
,
Ceren
Emir,
Gulen
Irem
Kaya,
Ahmet
Emir,
Mustafa
Ali
Onur
P393
Phenolic
compounds
of
Phlomis
samia
L.
from
Turkey
Göger
Fatih,
Köse
Yavuz
Bülent,
Kırımer
Neşe
P394
Investigation
of
polysaccharide
composition
in
medicinal
and
non-‐medicinal
aloes
Louise
I.
Ahl,
Henriette
L.
Pedersen,
William
G.
T.
Willats,
Nina
Rønsted,
Olwen
M.
Grace
P395
Spectral
and
chemical
studies
on
Magnoflorine
from
Zanthoxylum
armatum
DC
(Rutaceae)
leaves
and
evaluation
of
its
antistress
potential
Nitin
Verma,
R.
L.
Khosa
P396
Inhibition
of
interleukin-‐8
release
in
human
neutrophils
by
Melodorum
fruticosum
Nora
Engels,
Birgit
Waltenberger,
Barbara
Michalak,
Loi
Huynh,
Hung
Tran,
Anna
Kiss,
Hermann
Stuppner
P397
Isomer
of
bergenin
and
phytosterol
from
the
stem
bark
of
Mallotus
leucodermis
Aiza
Syuhada
Mohd
Yusoff,
Norizan
Ahmat,
Humera
Naz
P398
Flavones
from
the
twigs
of
Cynometra
cauliflora
Linn.
Nik
Fatini
Nik
Azmin,
Norizan
Ahmat
P399
Immunometabolic
regulation
by
triterpene-‐enriched
fraction
of
Eucalyptus
tereticornis
in
adi-‐
pose
tissue
cell
line
models.
Sergio
Acin,
Susana
Ceballos,
Diana
L.
Muñoz,
Adriana
Castaño,
Fernando
Echeverri,
Norman
Balcazar
P400
Evaluation
of
the
hypoglycemic
effects
of
extracts
and
diterpenoids
from
Azorella
compacta
(llareta)
Aurelio
San-‐Martín,
Mitchell
Bacho,
Sylvian
Cretton,
Philippe
Christen,
Andres
Olea,
Diana
Muñoz,
Alis
Guillen,
Norman
Balcazar
P401
New
sesquiterpene
lactones
from
Ambrosia
cumanensis
Kunth.
N.
Malafronte,
R.
Cotugno,
N.
Jimenez,
M.
De
Leo,
A.
Braca,
N.
De
Tommasi
P402
Antioxidant
capacity
of
Crataegus
monogyna
and
Crataegus
pentagyna
leaves
and
fruits
har-‐
vested
from
the
Danube
Delta
Oana
C.
Bujor,
Camelia
P.
Stefanache,
Irina
Volf,
Doina
Danila
P403
Sesquiterpene
lactones
and
phenolic
compounds
content
in
Arnica
montana
flowers
and
leaves
samples
harvested
from
wild
sites
in
North-‐East
Romania
Camelia
P.
Stefanache,
Oana
C.
Bujor,
Radu
Necula,
Valentin
Grigoras,
Constantin
Mardari,
Ciprian
Birsan,
Doina
Danila
P404
Investigation
of
potential
anti-‐diabetic
effect
of
Mucuna
pruriens
(L)
DC
(Fabaceae)
aqueous
leaf
extract.
Oke-‐Oghene
P.
Akpoveso,
Vesna
Tumbas-‐Saponjac,
Prabal
K.
Chatterjee,
George
Olivier
P405
Phytochemical
study
of
Helleborusodorus
subsp.
cyclophyllus
(Ranunculaceae)
Olga
St.
Tsiftsoglou,
Eirini
N.
Kalpourtzi,
Michalis
K.
Stefanakis,
Diamanto
M.
Lazari
P406
Study
of
Mezoneuron
benthamianum,
a
plant
traditionally
used
against
malaria
in
Guinea
Alembert
T.
Tchinda,
Jean
Loua,
Virginie
Esters,
Ewa
Cieckiewicz,
Allison
Ledoux,
Luc
Angenot,
Monique
Tits,
Aliou
M.
Balde,
Michel
Frédérich,
Olivia
Jansen
P407
In-‐vitro
bioassays
for
plant
extracts
with
tick-‐repellent
and
acaricidal
properties:
a
systematic
review
and
meta-‐analysis
Olubukola
Adenubi,
Folorunso
Fasina,
Lyndy
McGaw,
Jacobus
Eloff,
Vinny
Naidoo
P408
Toxicological
and
antimicrobial
studies
of
ethanolic
leaf
extract
of
Ricinodendron
heudelotii
(Baill.)
Heckel
Omolara
F.
Yakubu,
Abiodun
H.
Adebayo,
Olubanke
O.
Ogunlana,
Jacob
O.
Popoola,
Temitope
A.
Ishola,
Loretta
O.
Imonikhe,
Oladipupo
A.
Adeyemi
P409
Lupinifolin
extracted
from
Derris
reticulata
inhibits
growth
of
Staphylococcus
aureus
possibly
by
damaging
bacterial
cell
membrane
Kamol
Yusook,
Oratai
Weeranantanapan,
Siriporn
Riyajan,
Jidapa
Musika,
Nuannoi
Chudapongse
P410
Evolution
with
age
of
main
bitter
compounds
in
the
roots
of
cultivated
Gentiana
lutea
subsp.
aurantiaca
Óscar
González-‐López,
Sara
Mayo,
Álvaro
Rodríguez-‐González,
Carro,
Guzmán,
Paulo
H.
da
Silva,
Pedro
A.
Casquero
P411
Antiproliferative
diterpenoids
from
the
roots
of
Podocarpus
neriifolius
P.
Annécie
Benatrehina,
Tran
Ngoc
Ninh,
Carla
Slebodnick,
Djaja
Djendoel
Soejarto,
L.
Harinantenaina
Rakotondraibe,
A.
Douglas
Kinghorn
P412
Rhizome
weight
and
bioactive
compounds
of
Dioscorea
birmanica under
different
shading
Panumart
Rithichai,
Yaowapha
Jirakiattikul,
Piyapat
Khemwichai,
Srisopa
Ruangnoo,
Arunporn
Itharat
P413
Antiplasmodial
activity
and
identification
of
a
new
iridoid
triacetate
from
Heinsia
crinata
Tshisekedi
T.
P.,
Kalenda
T.
D.,
Mutwale
K.
P.,
Cieckiewicz
E.,
Jansen
O.,
Angenot
L.,
Tits
M.,
Frédérich
M.
P414
Evaluation
of
in
vitro
activity
and
ultrastructural
changes
in
Leishmania
amazonensis
caused
by
sesquiterpene
lactones
from
Calea
pinnatifida
(Asteraceae)
Patricia
Sartorelli,
Meire
L.
Yoshinaga,
Marcelo
José
P.
Ferreira,
João
Henrique
G.
Lago1,
Luiz
Felipe
D.
Passero
P415
Endemic
Asteraceae
from
Madeira
archipelago:
A
relation
of
hypoglycemic
activity
to
their
pol-‐
yphenolic
composition
Vítor
Spínola,
Sandra
Gouveia-‐Figueira,
Paula
C.
Castilho
P416
4-‐Methyl-‐1-‐alkylresorcinols,
novel
chemotaxonomic
markers
from
the
Spanish
medicinal
plant
Mercurialis
tomentosa
L.
Peter
Lorenz,
Miriam
Heinrich,
Dietmar
R.
Kammerer,
Florian
C.
Stintzing
P417
Assessment
of
Rauvolfia
nukuhivensis:
from
uses
to
biological
ac-‐tivities
and
chemodiversity
Nicolas
Martin,
Sara
F.
Ferreiro,
Eva
Alonso-‐Lopes,
Soizic
Prado,
Florent
Barbault,
Maël
Nicolas,
Gaël
Lecellier,
Christian
Paetz,
Marc
Gaysinski,
Olivier
Thomas,
Luis
Botana,
Phila
Raharivelomanana
P418
Anti-‐ageing
activity
of
Fitchia
nutans
extract,
a
Polynesian
tradi-‐tional
monoï
skin
care
ingredi-‐
ent
Jean-‐Luc
Ansel,
Quoc
Ly,
Jean-‐François
Butaud,
Mael
Nicolas,
Gaëtan
Herbette,
Laurent
Peno-‐
Mazzarino,
Elian
Lati,
Phila
Raharivelomanana
P419
Anti-‐fibrotic
effects
of
Silimarit®
dry
extract
in
the
STAM™
model
of
non-‐alcoholic
steatohepati-‐
tis
Philipp
Peterburs,
Aldo
Ammendola,
Anja
Lechner
P420
In
vitro
and
cellular
antioxidant
activities
of
Antidesma
thwaitesianum
Müll.
Arg.
(Euphorbi-‐
aceae)
leaf
extracts
Pintusorn
Hansakul,
Bhanuz
Dechayont,
Nitra
Nuengchamnong,
Arunporn
Itharat
P421
Cytotoxic
activity
against
liver
cancer
cell
[HepG2]
of
a
Thai
traditional
remedy
used
for
liver
cancer
treatment
Ponlawat
Maki,
Thammarat
Tui-‐on,
Arunporn
Itharat
P422
Effects
of
biotransformation
of
prenylated
flavonoids
on
antioxidative
capacity
Yina
Xiao,
Ik-‐Soo
Lee
P423
From
traditional
uses
to
phytochemical
study
of
a
Polynesian
healing
plant:
Constituents
and
properties
of
“Metuapua’a”
Raimana
Ho,
Taivini
Teai,
Jean-‐Pierre
Girault,
Alain
Meybeck,
René
Lafont,
Phila
Raharivelomanana
P424
Antiprotozoal
activity
against
Entamoeba
histolytica
of
furocoumarins
isolated
from
Ruta
cha-‐
lepensis
Ramiro
Quintanilla-‐Licea,
Benito
D.
Mata-‐Cárdenas,
Javier
Vargas-‐Villarreal,
Aldo
F.
Bazaldúa-‐
Rodríguez,
María
J.
Verde-‐Star
P425
A
new
inhibitor
of
hepatitis
C
virus
replication
from
Juncus
maritimus,
a
Tunisian
extremophile
plant
Ramla
Sahli,
Céline
Rivière,
Marie-‐Emmanuelle
Sahuc,
Smaoui
Abderrazak,
Jennifer
Samaillie,
Vincent
Roumy,
Thierry
Hennebelle,
Yves
Rouillé,
Karin
Seron,
Sevser
Sahpaz,
Riadh
Ksouri
P426
Chemical
composition,
anti-‐inflammatory
and
antioxidant
activity
activities
of
essential
oil
of
Piper
cubeba
L.
Ramzi
Mothana,
Mansour
AlSaid,
Mohammad
Raish,
Mohammed
Al-‐Sohaibani,
Ajaz
Ahmad,
Moham-‐
med
Al-‐Yahya,
Jamal
Kaled,
Syed
Rafatullah
P427
Identification
of
phenolic
and
volatile
compounds
in
Centhranthus
longiflorus
subsp.
longiflorus
by
HPLC
and
GC-‐MS,
antioxidant
and
antimicrobial
activity
studies
Sıla
Özlem
Şener,
Merve
Badem,
Nuriye
Korkmaz,
Rezzan
Aliyazicioglu,
Ufuk
Özgen,
Şengül
Alpay
Ka-‐
raoğlu
P428
Phenolic
composition
by
RP-‐HPLC
and
antioxidant
capacity
studies
of
Consolida
orientalis
(Gay)
Schrod.
Rezzan
Aliyazicioglu,
Ufuk
Özgen,
Merve
Badem,
Sıla
Özlem
Şener,
Nuriye
Korkmaz,
Şengül
Alpay
Karaoğlu
P429
HPLC
Profiles
of
phenolic
compounds,
antioxidant
and
antimicrobial
activity
of
the
aerial
parts
of
Coronilla
varia
L.
Rezzan
Aliyazicioglu,
Ufuk
Özgen,
Sıla
Özlem
Şener,
Merve
Badem,
Nuriye
Korkmaz,
Şengül
Alpay
Karaoğlu
P430
Antimicrobial
and
antioxidant
activities
of
the
aerial
parts
and
roots
of
Cirsium
trachylepis
Rezzan
Aliyazicioglu,
Ufuk
Özgen,
Sıla
Özlem
Şener,
Merve
Badem,
Şengül
Alpay
Karaoğlu
P431
Evaluation
and
isolation
of
antimutagenic
compounds
from
methanolic
leaf
extracts
of
Monan-‐
thotaxis
caffra
Rhulani
Makhuvele,
Kenn
Foubert,
Sandra
Apers,
Luc
Pieters,
Luc
Verschaeve,
Esameldin
E.
Elgorashi
P432
Potential
role
of
arbuscular
mycorrhizal
fungi
in
the
accumulation
of
polyphenols
in
Lamiaceae
species
Rita
Engel,
Krisztina
Szabó,
László
Abrankó,
Anna
Füzy,
Tünde
Takács
P433
Chemical
composition
and
bioactivity
of
the
essential
oil
of
Pinus
roxburghii
bark
Rola
M.
Labib
,
Fadia
S.
Youssef,
Mohamed
L.
Ashour,
Jennifer
Búfalo,
Samir
A.
Ross
P434
Investigations
in
flavonoid
content
of
Camelina
sativa
(L.)
Crantz
during
seed
development
Roland
Molinié,
Anthony
Quéro,
David
Mathiron,
Benjamin
Thiombiano,
Jean-‐Xavier
Fontaine,
Déborah
Brancourt,
Olivier
Van
Wuytswinkel,
Emmanuel
Petit,
Hervé
Demailly,
Gaëlle
Mongelard,
Serge
Pilard,
Brigitte
Thomasset,
François
Mesnard
P435
Greek
flora
as
a
source
of
new
anti-‐oxidant,
anti-‐elastase,
anti-‐collagenase
and
anti-‐
hyaluronidase
natural
agents
Michalea
Rozalia,
Boka
Vasiliki-‐Ioanna,
Dina
Evanthia,
Aligiannis
Nektarios,
Skaltsounis
Alexios-‐
Leandros
P436
Search
for
leishmanicidal
agents
by
bioactivity-‐correlated
techniques
from
terrestrial
plants
Manisha
Bhatnagar,
Nandan
Sarkar,
Sabari
Ghosal
P437
Antiprotozoal
and
heme-‐binding
activity
of
13
aromatic
and
medicinal
plants
from
Ahaggar,
Algeria
Kamel
Dali-‐Yahia,
Sergio
Ortiz,
Alexandre
Maciuk,
Pedro
Vásquez-‐Ocmín,
Laila
Salmen
Espindola,
Sa-‐
brina
Boutefnouchet
P438
Chemical
characterization
of
Peganum
harmala
seeds
oil
with
evaluation
of
some
biological
activities
Khadhr
M,
Bousta
M,
Boukhira
S,
Boukhchina
Sadok
P439
Anti
freckles
plants
in
Iranian
Traditional
Medicine
Saeedeh
Ghafari,
Shirin
Fahimi,
Shamim
Sahranavard,
Ghazaleh
Heydarirad
P440
Alkaloids
of
Narcissus
poeticus
cv.
Pink
Parasol
and
their
biological
activity
Šafratová
Marcela,
Hošťálková
Anna,
Opletal
Lubomír,
Kuneš
Jiří,
Cahlíková
Lucie
P441
Bioactive
labdane
diterpenoids
from
Salvia
leriifolia
Samad
Nejad
Ebrahimi,
Mahdi
Moridi
Farimani,
Akram
Taleghani,
Abbas
Aliabadi,
Atousa
Aliahmadi,
Mohammad
Ali
Esmaeili,
Nazanin
Namazi
Sarvestani,
Hamid
Reza
Khavasi,
Martin
Smieško,
Matthias
Hamburger
P442
Resistance
modulatory
and
efflux
pump
inhibitory
activities
of
Kaempferia
galanga
rhizomes
Sandra
Prasch,
Britta
Fimbinger,
Franz
Bucar
P443
Novel
polyketides
isolated
from
cultures
of
an
endophytic
fungus,
Annulohypoxylon
truncatum
Wei
Li,
Changyeol
Lee,
Sung
Hee
Bang,
Soonok
Kim,
Xuikui
Xia,
Sang
Hee
Shim
P444
Anti-‐fatigue
effects
of
Rhodiola
rosea
extract
Sang
Yoon
Choi
P445
Vaccinium
cereum
leaves,
Tahitian
berry,
a
common
endemic
plant
with
α-‐glucosidase
inhibito-‐
ry
activity
Sarah
Benayad,
Estelle
Adam,
Stéphanie
Soulet,
Isabelle
Bombarda,
Cécile
Debitus,
Taivini
Teai
P446
New
flavonoid
from
Polygonum
equesitiform
Sm.
and
antioxidant
activity
of
its
extract
Sayed
A.
El-‐Toumy,
Josline
Y.
Salib,
Nabila
H.
Shafik,
Asmaa
Sayed
P447
Isolation
of
flavonoids
from
Acacia
albida
leaves
and
evaluation
of
antihyperglycaemic
effects
of
its
extract
Sayed
A.
El-‐Toumy,
Ahmed
H.
Gaara,
Abdel
Razik
H.
Farrag,
Nadia
M.
Ahmed
P448
Physiological
activities
of
Rubus
crataegifolius
fruits
in
Korea
Hyeusoo
Kim,
Moon-‐Sup
Kim,
Sugwang
Lee,
Uk
Lee,
Sea-‐Hyun
Kim
P449
Antioxidant
activities
and
useful
components
of
Korean
Walnut
(Juglans
sinensis)
kernels
Youngki
Park,
Sea-‐Hyun
Kim,
Jae-‐Hee
Kim
P450
Phytochemical
analysis
of
aerial
parts
of
Persicaria
bistorta
(L.)
Samp.
and
the
evaluation
of
the
anti-‐inflammatory
potential
of
rare
malonylated
flavonols.
Urszula
Klimczak,
Marta
Woźniak,
Michał
Tomczyk,
Sebastian
Granica
P451
Chemical
composition
of
Prangos
hulusii
Secil
Yazici-‐Tutunis,
Nur
Tan,
Mahmut
Miski
P452
In
vitro
antidiabetic
activity
of
Cotinus
coggygria
Scop.
Sefa
Gözcü,
Hafize
Yuca,
Benan
Dursunoğlu,
Zühal
Güvenalp,
L.
Ömür
Demirezer
P453
Neuroprotective
effect
of
Perilla
extracts
on
PC12
cells
Senavong Pimolvan, Kongkham S, Saelim S, Suangkavathin V
P454 Four new neolignans from the fruits of Juglans mandshurica Maxim.
SeonJu Park, Nanyoung Kim, Guijae Yoo, Jun Hyung Park, Seung Hyun Kim
P455 Heterocyclic compounds from the fruits of Morus alba L. and their anti-‐angiogenesis activities
Seoung Rak Lee, Seulah Lee, Hee Jeong Eom, Hee Rae Kang, Jae Sik Yu, Tae Kyoung Lee, Jiwon Baek,
Dahae Lee, Won Se Suh, Ki Hyun Kim
P456 Essential oils from Artemisia copa (Asteraceae) and Aloysia deserticola, two medicinal species
used in the Chilean Atacama’s community (Antofagasta)
S. Ortiz, Pierre Champy, S. Boutefnouchet
P457 Determination of phenolic compounds of Tribulus terrestris (Zygophyllaceae)
Sevim Küçük, Fatih Göger, Neşe Kırımer
P458 Promising Neuroprotective and Anti-‐Inflammatory Effect of Small Nettle’ Ethyl Acetate Extract
Sevki ARSLAN, Gulsum TERZIOGLU, Buket Kabalay, Ali Riza TUFEKCI, Alaattin SEN
P459 Antiviral and antibacterial effects of 16 plants used in Ivorian traditional medicine
Moussa Bamba, Simon Bordage, Marie-‐Emmanuelle Sahuc, Jennifer Samaillie, Christel Neut, Alexis
Zamble, Fezan Tra Bi, Karin Seron, Sevser Sahpaz
P460 Antimicrobial and acetylcholinesterase inhibition activities of an endemic plant from Sahara:
Myrtus nivellei Batt. & Trab. (Myrtaceae)
Rym G. Demmak, Simon Bordage, Christel Neut, Abderahman Bensegueni, Sevser Sahpaz
P461 Effect of Crocus sativus, as an accompaniment agent in traditional Persian medicine, on pharma-‐
cokinetic of acetaminophen
Seyede Nargess Sadati, Sima Sadrai, Mohammad R. Shams Ardekani, Katayoon Mireskandari, Moham-‐
mad Sharifzadeh
P462 Quantitative analysis of phenolic compounds of some Crataegus species from Turkey
Sezin ANIL, Esra EROĞLU ÖZKAN, Ali DÖNMEZ, Gülay MELİKOĞLU
P463 Cytoprotective and antiadhesive effects of aqueous leaf extract from Orthosiphon aristatus
against uropathogenic E. coli
Shabnam Sarshar, Mohammad R. Asadi Karam, Mehri Habibi, Saeid Bouzari, Xiaofei Qin, Francisco M.
Goycoolea, Simone Brandt, Andreas Hensel
P464 New cytotoxic constituents from Xestospongia testudinaria sponge collected from the Red Sea of
Saudi Arabia
Ali A. El-‐Gamal, Shaza M. Al-‐Massarani, Diaa T. A. Youssef, Lamiaa A. Shaala, Mansour S. Al-‐Said, Abdel-‐
kader E. Ashour, Ashok Kumar, Maged S. Abdel-‐Kader, Wael M. Abdel-‐Mageed
P465 Screening of compounds of Evernia prunastri (L.) for their antiproliferative activity in
glioblastoma cells
Shcherbakova A, Nyugen L, Koptina A, Backlund A, Shurgin A, Romanov E, Ulrich-‐Merzenich G
P466 Isolation and characterization of lupane-‐type triterpenes from Careya arborea and their effect
on modulation of pro-‐inflammatory mediators
Rayhana Begum, Manju Sharma, Showkat R. Mir
P467 Cytotoxic ambuic acid derivatives from Hawaiian endophytic fungus FT-‐172
Chun-‐Shun Li, Bao-‐Jun Yang, James Turkson, Shugeng Cao
P468 Alpine plants with anti-‐ageing effects
Silvia Revoltella, Birgit Waltenberger, Giorgia Baraldo, Clemens Kohl, Pagitz Konrad, Roland Kohl, Pid-‐
der Jansen-‐Dürr, Hermann Stuppner
P469 Triterpensaponins as a novel transfection enhancer
Simko Sama, Matthias F. Melzig, Alexander Weng
P470 Autoxidation of Betulonaldehyde
Sloan Ayers, Tamas Benkovics, Jonathan Marshall, Yichen Tan
P471 Laricifomes officinalis – a rich source of pharmacologically active triterpenes
Sonja Sturm, Katharina Gallmetzer, Andreas Friedl, Birgit Waltenberger, Veronica Temml, Hermann
Stuppner
P472
Anti-‐amyloidogenic
constituents
from
roots
of
Dryopteris
crassirhizoma
in
Alzheimer’s
disease
cellular
model
Hwan-‐Bin
Joo,
Jae-‐Moon
Kim,
So-‐Young
Park
P473
Anti-‐inflammatory
activity
of
a
Carica
papaya
leaf
extract
used
to
prevent
occasional
gingivitis
Stéphane
Dejoie,
Rim
Boulmane,
Séverine
Derbré,
Daniel
Henrion,
Christophe
Binachon,
Pascal
Ri-‐
chomme
P474
Potential
immunomodulatory
compounds
isolated
from
red
betel
leaves
(Piper
crocatum,
Ruiz
&
Pav.)
Subagus
Wahyuono,
Paula
Kustiawan,
Yustina
Sri
Hartini,
Yuswanto,
Sitarina
Widyarini
P475
The
potential
of
Combretum
molle
in
the
treatment
of
skin
cancer
Sunelle
Rademan,
Namrita
Lall
P476
A
bioactivity-‐guided
screening
of
Sri
Lankan
Plants
in
the
search
for
novel
antibacterial
and
anticancer
agents.
Supun
Mohotti,
Sanjeevan
Rajendran,
Taj
Muhammad,
Adam
A.
Strömstedt,
Robert
Burman,
Björn
Hellman,
E.D.
de
Silva,
Ulf
Göransson,
C.M.
Hettiarachchi,
Sunithi
Gunasekara
P477
Identifying
Specific
Inhibitors
of
Triple-‐Negative
Breast
Cancer
Subtypes
Andrew
J.
Robles,
Shengxin
Cai,
Lin
Du,
Corena
V.
Shaffer,
Tanja
Grkovic,
April
L.
Risinger,
Barry
R.
O’Keefe,
Robert
H.
Cichewicz,
Susan
L.
Mooberry
P478
Triterpene
saponin
constituents
from
roots
of
Bupleurum
falcatum:
Hepatoprotective
effects
on
D-‐galactosamine-‐induced
cell
damage
Takuya
Konno,
Kiyofumi
Ninomiya,
Masayuki
Yoshikawa,
Hisashi
Matsuda,
Toshio
Morikawa
P479
Multidrug-‐efflux
inhibitors
–
A
study
on
selected
medicinal
plants
Tariq
M.
Aljarba,
Paul
Stapleton,
Simon
Gibbons
P480
Selected
medicinal
plant
extracts
as
inhibitors
of
conjugal
transfer
of
plasmid-‐mediated
antibiotic
re-‐
sistance
in
Escherichia
coli
Tariq
M.
Aljarba,
Paul
Stapleton,
Simon
Gibbons
P481
A
new
strategy
for
the
evaluation
of
biological
properties
of
lipophilic
natural
products:
Appli-‐
cation
to
the
wound
healing
and
antibacterial
activities
of
Calophyllum
inophyllum
Linn
oil
Teddy
Léguillier,
Marylin
Lecsö-‐Bornet,
Christelle
Lémus,
Nicolas
Lebouvier,
Edouard
Hnawia,
Mo-‐
hammed
Nour,
William
Aalbersberg,
Phila
Raharivelomanana,
Patrice
Rat
P482
Arginase
inhibitors:
from
chlorogenic
acid
to
cinnamides
Thanh-‐Nhat
Pham,
Duc-‐Thu
Trinh,
Simon
Bordage,
Céline
Demougeot,
Marc
Pudlo,
Khac-‐Minh
Thai,
Corine
Girard-‐Thernier
P483
Anticonvulsant
agents
from
Boswellia
sacra
identified
by
zebrafish
bioassay-‐guided
fractiona-‐
tion
Théo
Brillatz,
Emerson
Ferreira
Queiroz,
Laurence
Marcourt,
Maxime
Jacmin,
Alexander
D.
Crawford,
Jean-‐Luc
Wolfender
P484
Bioguided
isolation
of
anticonvulsant
principles
from
Helleborus
cyclophyllus
using
the
zebrafish
epilepsy
model
Théo
Brillatz,
Emerson
Ferreira
Queiroz,
Laurence
Marcourt,
Konstantina
Vougogiannopoulou,
Maxime
Jacmin,
Alexander
D.
Crawford,
Leandros
Skaltsounis,
Jean-‐Luc
Wolfender
P485
New
compounds
from
the
leaves
of
Cleistocalyx
operculatus
and
their
inhibitory
activities
on
influenza
A
neuraminidases
Thi-‐Kim-‐Quy
Ha,
Won
Keun
Oh
P486
Antifungal
activities
and
chemical
composition
of
the
essential
oil
of
Lippia
micromera
(Verbe-‐
naceae)
cultivated
in
French
Guiana
Camille
Scotto,
Pauline
Burger,
Thomas
Michel,
Mehdi
Khodjet
el
khil,
Marine
Ginouves,
Ghislaine
Prevot,
Denis
Blanchet,
Piero
G.
Delprete,
Xavier
Fernandez
P487
Anti-‐inflammatory,
cytotoxic
and
antimicrobial
activities
of
Piper
peltatum
leaf
extract
Thomas
Michel,
Audrey
Kerdudo,
Emy
Njoh
Ellong,
Vanessa
Gonnot,
Stéphane
Rocchi,
Jean-‐François
Tanti,
Laurent
Boyer,
Sandra
Adenet,
Katia
Rochefort,
Xavier
Fernandez
P488
Dragonbloodin
A1
and
A2:
novel
flavan
trimers
and
anti-‐inflammatory
principles
from
Sanguis
Draconis
Wen-‐Ke Du, Hsin-‐Yi Hung, Ping-‐Chung Kuo, Tian-‐Shung Wu
P489 Two new acylated flavonol glycosides from Mimosa pigra leaves
Chinedu J. Okonkwo, Tochukwu J. N. Okonkwo, Ozadheoghene E. Afieroho, Blessing O. Okonkwo,
Charles O. Esimone, Obioma U. Njoku, Peter Proksch
P490 Comparison of genotoxicity potentials between crude Thai bee pollen and its extracts
Treetip Ratanavalachai, Sumon Thitiorul, Wantha Jenkhetkan, Chalerm Jansom, Arunporn Itharat
P491 Antimutagenic activity of n-‐tetracosanol, eicosanoic acid and arjunolic acid isolated from Com-‐
bretum microphyllum (Combretaceae)
Tshepiso J. Makhafola, Esameldin E. Elgorashi, Lyndy J. McGaw, Maurice D. Awouafack, Luc Verschaeve,
Jacobus N. Eloff
P492 Evaluation for the anti-‐leukemic activities and 28-‐day subacute toxicity of ethanolic extracts
from the fruiting bodies of dish-‐cultured Antrodia cinnamomea
Tung-‐Ying Wu, Mohamed El-‐Shazly, Ying-‐Chi Du, Yu-‐Ming Hsu, Kuei-‐Hung Lai, Mei-‐Chin Lu, Yang-‐Chang
Wu
P493 Bioactive constituents from the termite nest-‐derived medicinal fungus Xylaria nigripes
Jung-‐Chun Chang, George Hsiao, Ruo-‐Kai Lin, Yueh-‐Hsiung Kuo, Yu-‐Min Ju, Tzong-‐Huei Lee
P494 Phenolic compounds of Ornithogalum pyrenaicum together with antimicrobial and antioxidant
studies
Nuriye Korkmaz , Sıla Özlem Şener, Merve Badem, Ufuk Özgen, Rezzan Aliyazicioglu, Şengül Alpay Ka-‐
raoğlu
P495 Bioactivity studies of algal extracts from Turkey’s Mediterranean coast
Yasin Genc, Secil Sarikaya Aydin, Melek Sertdemir, Ozge Demir, Emine S. Okudan, Selin Salin, Belma
Konuklugil, Ummuhan Sebnem Harput
P496 Bioassay guided isolation studies on Incarvillea emodi (Royle ex Lindl.) Chatterjee
Yasir Ihtesham, Zeynep Dogan, Vahap Murat Kutluay, Uzma Khan, Iclal Saracoglu
P497 Cytotoxicity comparison on different parts of three Digitalis species and isolation of potential
bioactive compounds
Vahap Murat Kutluay, Makoto Inoue, U. Sebnem Harput, Iclal Saracoglu
P498 Quantitative analysis of polyphenols and antioxidant activity of Croatian populations of Erodi-‐
um cicutarium (L.) L'Herit (Geraniaceae)
Vanja Ljoljić Bilić, Jadranka Vuković Rodríguez, Renata Jurišić Grubešić, Dario Kremer, Živka Juričić
P499 Secondary metabolites in honey and their inhibitory activity on matrix metalloproteinases
Boznou Vasiliki, Polychronopoulos Panagiotis, Michalis Zervos, Stavropoulou Maria-‐Ioanna, Aligiannis
Nektarios
P500 Studies on the chemical stability of umbelliprenin, the active principle of Ferula spp.
Vito Alessandro Taddeo, Francesco Epifano, Philippe de Medina, Serena Fiorito, Salvatore Genovese
P501 Discovery of novel protein tyrosine phosphatase 1B inhibitors from medicinal plant resources
Wei Li, Tatsunori Sasaki, Toshihisa Onoda, Koji Higai, Kazuo Koike
P502 Structure characterization and immunomodulating effects of polysaccharides isolated from
Dendrobium officinale
Wei Wei, Lei Feng, Shao-‐Ping Nie, and Quan-‐Bin Han
P503 SAR study and antitumor potential of plant natural product miliusanes and their derivatives
Wen-‐Hui Pan, Yi-‐Fu Guan, Kang-‐Lun Liu, Xin-‐Ya Xu, Xun Song, Dong-‐Ying Wang, Wen-‐Jian Xie, Siu Wai
Tsang, Hong-‐Jie Zhang
P504 Cytotoxicity effects and apoptosis induction by cycleanine and tetrandrine
Fidelia I. Uche, Falko Drijfhout, James McCullagh, Alan Richardson, Wen-‐Wu Li
P505 Antibacterial effects of the essential oil from flower buds of Magnolia biondii Pamp
Omar Aldulaimi, Wen-‐Wu Li
P506 Antimicrobial and antioxidant efficacy of Acokanthera oblongifolia (Apocynaceae) used for skin
diseases by Xhosa tribe of Amathole District, Eastern Cape, South Africa
Wilfred M. Otang, Anthony J. Afolayan
P507
Chemical
constituents
from
Melicope
pteleifolia
leaves
and
their
anti-‐influenza
virus
activities
Ngoc-‐Hieu
Nguyen,
Thi-‐Kim-‐Quy
Ha,
Won
Keun
Oh
P508
Antitumor
activity
of
periplocin,
isolated
from
Telectadium
dongnaiense,
in
colorectal
cancer
cells
via
suppression
of
Wnt/β-‐catenin
signaling
Won
Kyung
Kim,
Ba-‐duc
Hiep,
Tae
Joon
Choi,
Je
Do
Oh,
Hyen
Joo
Park,
Sei
Ryang
Oh,
Sang
Kook
Lee
P509
Cytotoxic
triterpenes
from
the
twigs
of
Chaenomeles
sinensis
Won
Se
Suh,
Chung
Sub
Kim,
Kyoung
Jin
Park,
Joon
Min
Cha,
Oh
Kil
Kwon,
Sang
Un
Choi,
Kang
Ro
Lee
P510
Four
new
flavonoid
glycosides
from
the
white
flower
of
Impatiens
balsamina
Won
Se
Suh,
Chung
Sub
Kim,
Kyoung
Jin
Park,
Joon
Min
Cha,
Oh
Kil
Kwon,
Sang
Zin
Choi,
Mi
Won
Son,
Kang
Ro
Lee
P511
Bioactive
chemical
constituents
from
the
twigs
of
Salix
glandulosa
Won
Se
Suh,
Chung
Sub
Kim,
Kyoung
Jin
Park,
Joon
Min
Cha,
Dong
Hyun
Kim,
Kang
Ro
Lee
P512
Fatty
acids
from
the
stem
bark
of
Sorbus
commixta
and
their
biological
activity
Won
Se
Suh,
Chung
Sub
Kim,
Kyoung
Jin
Park,
Joon
Min
Cha,
Tae
Hyun
Lee,
Sun
Yeou
Kim,
Kang
Ro
Lee
P513
Antiviral
effect
of
Epimedium
koreanum
extract
Won-‐Kyung
Cho,
Jong-‐Soo
Lee,
and
Jin
Yeul
Ma
P514
New
Labdane
Diterpenoids
from
Dysoxylum
hongkongense
Yao-‐Ching
Tsai,
Tsong-‐Long
Hwang,
Yuh-‐Chi
Kuo,
Ching-‐Te
Chien,
Ya-‐Ching
Shen
P515
Anti-‐inflammatory
diterpenoids
from
Callicarpa
randaiensis
Ho-‐Hsi
Cheng,Yuan-‐Bin
Cheng,
Tsong-‐Long
Hwang,
Yao-‐Haur
Kuo,
Ya-‐Ching
Shen
P516
Inhibitory
effects
of
andaliman
essential
oil
on
acid
production,
volatile
sulfur
compounds,
and
Actinomyces
viscosus
biofilms
Yanti,
Berti
Priska
Gea,
Bibiana
W.
Lay,
Marco
Tjakra,
Stevhen
Juniardi
P517
Cucurbitane-‐type
triterpenes
and
glycoside
from
the
rattan
of
wild
Momordica
charantia
and
their
anti-‐inflammatory
and
cytotoxic
activities
Li-‐Jie
Zhang,
Hung-‐Tse
Huang,
Chia-‐Ching
Liaw,
Shih-‐Yen
Huang,
Zhi-‐Hu
Lin,
Yao-‐Haur
Kuo
P518
Effect
of
elicitors
on
bioactive
compound
accumulation
in
shoot
culture
of
Dioscorea
membra-‐
nacea
Yaowapha
Jirakiattikul,
Panumart
Rithichai,
Thipsukon
Boonyuen,
Srisopa
Ruangnoo,
Arunporn
Itharat
P519
Phenolic
compounds
of
Salvia
hypargeia
Fich.
&
Mey.
Yavuz
Bülent
Köse,
Fatih
Göger,
Gökalp
İşcan
P520
Medicinal
plants
traded
in
traditional
markets
of
southern
part
of
Guizhou,
Southwest
China
Liya
Hong,
Yizhou
Wang,
Jianqin
Li,
Qiyi
Lei,
Jiangju
Zhou,
Liang
Qin,
Chunlin
Long
P521
Comparing
the
leaf
secretory
apparatus
of
Hibiscus
sabdariffa
and
Hibiscus
surattensis
K.
Raghu,
Y.
Naidoo
P522
Soluble
epoxide
hydrolase
inhibitory
activity
of
anthraquinone
components
from
Aloe
Ya
Nan
Sun,
Ah
Reum
Jo,
Jang
Hoon
Kim,
Jong
Seong
Kang,
Young
Ho
Kim
P523
Lignans
with
inhibitory
activity
against
PCSK9
mRNA
expression
from
the
fruits
of
Schisandra
chinensis
Young-‐Mi
Kim,
Young
Hee
Choi,
Young-‐Won
Chin
P524
Chemical
constituents
and
bioactivities
of
the
twigs
of
Severinia
buxifolia
Yuan-‐Bin
Cheng,
Pei-‐Shian
Li,
Yang-‐Chang
Wu,
Fang-‐Rong
Chang
P525
Study
on
the
anti-‐melanoma
constituents
from
Pinus
taiwanensis
Hayata
Yu-‐Chen
Kao,
Jen-‐Der
Wu,
Ching-‐Kuo
Lee
P526
Anti-‐inflammatory
activity
of
resveratrol
metabolites
Yulia
Radko,
Steen
B.
Pedersen,
Lars
P.
Christensen
P527
Antimutagenic
potentials
of
flavonoids
from
Achillea
millefolium
L.
subsp.
millefolium
Handan
G.
Sevindik,
Zühal
Güvenalp,
Mehmet
Karadayı,
Medine
Güllüce,
L.
Ömür
Demirezer
P528
Active
compounds
from
Achillea
biebersteinii
and
determination
of
their
antigenotoxic
poten-‐
tials
Handan G. Sevindik, Zühal Güvenalp, Mehmet Karadayı, Medine Güllüce,
L. Ömür Demirezer
P529 Bauhinia variegata L.; a bullet against pathological changes behind cognitive impairment; phy-‐
tochemical, in vitro and in vivo study
Alaa Selim, Heba Handoussa, Nesrine El-‐Sayed
P530 Production of immature rhizomes by breaking dormancy under low temperature of Gastrodia
elata Blume
Changsu Kim, Dongwon Kim, Hyojin Kim, Youngju Song, Wanghyu Lee
P531 Gram-‐scale purification of faradiol from common marigold
Dezső Csupor, Szabina Izrael, Norbert Kúsz, Attila Horváth
P532 Anti-‐HIV1 potential of an endemic Brazilian bamboo species
Janayne Gagliano, Cláudia Maria Furlan
P533 Investigation of the differences between the “Cold” and “Hot” nature on immune endocrine me-‐
tabolism levels of Açaí (Euterpe oleracea Mart.)
Jianjun Zhang, Linyuan Wang, Chun Wang, Yingli Zhu, Zichen Wang, Yan Qu, Li Wu
P534 Evaluation of glucosinolate content in radish (Raphanus sp.) germplasm
Ho-‐Cheol Ko, Mun-‐Sup Yoon, On-‐Sook Hur, Jung-‐Sook Sung, Jung-‐Bong Kim, Jae-‐Gyun Gwak, Hyung-‐Jin
Baek, Sang-‐Gyu Kim, Binod P. Luitel, Kyoung-‐Yul Ryu and Ju-‐Hee Rhee
P535 Investigation of polysaccharide composition in medicinal and non-‐medicinal aloes
Louise I. Ahl, Henriette L. Pedersen, William G. T. Willats, Nina Rønsted, Olwen M. Grace
P536 New insights into anti-‐S. aureus action of Buchenavia tetraphylla and Libidibia ferrea: inhibition
of DNA replication
Luís Cláudio Nascimento da Silva, José Robson Neves Cavalcanti Filho, Clovis Macedo Bezerra Filho,
Tiago Fonseca da Silva, Márcia Vanusa da Silva, Maria Tereza dos Santos Correia, Anders Løbner-‐Olesen
P537 Steroidal saponins from Chlorophytum deistelianum
Turibio Tabopda, Anne-‐Claire Mitaine-‐Offer, Thomas Paululat, Stéphanie Delemasure, Patrick Dutartre,
Bonaventure Tchaleu Ngadjui, Marie-‐Aleth Lacaille-‐Dubois
P538 The use of medicinal plants grown hydroponically in the treatment of wounds
Maxim Timoshenko , Natalya Voroshilova1, Marlen Yessirkepov, Assilbek Burabaev
P539 Usefulness of Φ-‐order kinetics for the investigation, characterisation and quantification of pho-‐
todegradation processes.
Mounir Maafi, Mohammed Alqarni
P540 Mechanism(s) of action involved in the antimycobacterial activity of Tetracera macrophylla
Siti Fatimah Sabran, Maryati Mohamed, Mohd Fadzelly Abu Bakar, Alona Cuevas Linatoc
P541 Bioassay guided isolation studies on Scutellaria salviifolia Benth.
Zeynep Dogan, V. Murat Kutluay, Iclal Saracoglu
P542 Compounds from the aerial parts of Agrimonia pilosa and their protein tyrosine phosphatase 1B
and acetylcholinesterase inhibitory activities
Duc H. Nguyen, U M. Seo, Duc D. Le, Thi T. Nguyen, Jae Sue Choi, Mi H. Woo
P543 Evaluation of 5-‐HT1A and 5-‐HT2A receptors in a rat model of chronic restraint stress: antidepres-‐
sant-‐like effects of albiflorin
Yingli Zhu, Linyuan Wang, Jiangxia Wang, Zhihui Yang, Chenglong Wang, Danping Zhao, Jianjun Zhang
P544 Inhibitory effects of epimedium herb water extract on the inflammatory response in vitro and in
vivo
You-‐Chang Oh, Yun Hee Jeong, Won-‐Kyung Cho, Sang-‐Joon Lee, Jin Yeul Ma
P545 LC-‐MS profiling, isolation and identification of bioactive secondary metabolites from a marine-‐
derived fungus Arthrinium sp.
Ahmed M. Elissawy, Sherif S. Ebada, Mohamed L. Ashour, Ferhat C. Özkaya, Weaam Ebrahim, Mohamed
S. ElNaggar, Peter Proksch, Abdelnasser Singab
P546 Marine-‐derived Aspergillus species are promising source of new secondary metabolome
Ala Ud Din, Wayne D. Inman, Frederick A. Valeriote, Phillip Crews
P547 Analgesic activity of Carthamus caeruleus ethanolic extracts
Amel Toubane, Kamel Daoud
P548 Biological active natural products from marine organisms and terrestrial plants of Iran
Amir Reza Jassbi
P549 The toxicity of ribbon worms: Alpha-‐nemertides or tetrodotoxin, or both?
H.S. Andersson, E. Jacobsson, C. Eriksson, M. Hedström, H. Seth, P. Sundberg, K.J. Rosengren, M. Strand,
U. Göransson
P550 Mo-‐clay for treatment of psoriasis
Kirstine M Hansen, Anna K Jäger, Åsa Andersson
P551 Determination of the antimicrobial and antitussive activities of the leaves of Solanum torvum
Abena Amponsaa Brobbey, Anna Kwarley Quartey, Samuel Otuo-‐Serebour, Isaac Ayensu
P552 Commercializing traditional knowledge: Regulating access and benefit-‐sharing in Namibia
Anne Heeren, Ahmad Cheikyoussef, Percy Chimwamurombe
P553 Oxymatrine and matrine biotransformation by microorganisms and the evaluation of effects on
5-‐lipoxygenase inhibition
Fatih Demirci, Ayşe Esra Karadağ, Mustafa Güzel
P554 Chemical and genetic differences between Hawaiian lineages of the alga Asparagopsis taxiformis
Benjamin R. Clark, Mindy Mizobe, Jo-‐Ann Leong, Robert P. Borris
P555 Antioxidant and antitumor studies of Phyllophora pseudoceranoides extracts
Adriana Trifan, Laura Bucur, Daciana Sava, Camelia Paula Stefanache, Ana Clara Aprotosoaie, Oana
Cioanca, Monica Hancianu, Anca Miron
P556 Preparative separation of marine bioactive compounds by centrifugal partition chromatog-‐
raphy
Romain Boulho, Julie Le Roux, Céline Le Quemener, Anne-‐Sophie Burlot, Grégoire Audo, Nathalie Bour-‐
gougnon, Gilles Bedoux
P557 Caulerpenyne from Caulerpa Taxifolia: a comparative study between CPC and classical chroma-‐
tographic techniques
Estelle Sfecci, Céline le Quémener, Grégoire Audo, Thierry Lacour, Philippe Amade, Mohamed Mehiri
P558 Study on the antibacterial components from Palythoa-‐symbiotic fungus, Fusarium solani
Chih-‐Chuang Liaw, Po-‐Yi Cheng
P559 Metabolite profiling of Baltic blue mussel (Mytilus sp.) hybrids and their associated microbes
Caroline Utermann, Corinna Breusing, Heiko Stuckas, Tim Staufenberger, Antje Labes, Deniz Tasdemir
P560 Chemical and biological screening of deep-‐water sponges from Antarctic regions
Fengie Li, Michael Marner, Arlette Wenzel-‐Storjohann, Johanna Silber, Dorte Janussen, Deniz Tasdemir
P561 Characterization of bioactive molecules derived from marine microorganisms, interfering with
cell dedifferentiations observed at the invasive front of mammary tumors
Ambre Dezaire, Nathalie Ferrand, Marine Vallet, Michèle Sabbah, Annette K. Larsen, Soizic Prado, Ale-‐
xandre Escargueil
P562 Botryane sesquiterpenes and binaphthalene tetrols from endophytic fungi associated to the
marine red algae Asparagopsis taxiformis
Rebeca P. Medina, Airton D. Silva, Raymond J. Andersen, Angela R. Araújo, Dulce H. S. Silva
P563 Cytotoxic compounds of sponges collected from West Bali National Park Indonesia
Erna Prawita Setyowati, Anggara Jenie, Sudarsono, Subagus Wahyuono
P564 P-‐Sulfooxyphenylpyruvic acid derivatives from the green macroalga Caulerpa taxifolia
Estelle Sfecci, Lionel Massi, Thierry Lacour, Philippe Amade, Mohamed Mehiri
P565 Bioassay-‐guided identification of bioactive protein compounds from the Mediterranean
demosponge Tethya meloni
Eugenia Gentile, Carlotta Nonnis Marzano, Tiziana Latronico, Anna Fasano, Rocco Rossano, Giuseppe
Corriero, Grazia Maria Liuzzi
P566 Oxymatrine and matrine biotransformation by microorganisms and the evaluation of effects on
5-‐lipoxygenase inhibition
Fatih Demirci, Ayşe Esra Karadağ, Mustafa Güzel
P567 The effects of brown macro algae from Northen coasts of Persian Gulf on melanogenesis
Foroogh Namjoyan, Mojtaba Alishahi, Alireza Jahangiri, Massoumeh Farasat, Hamideh Mousavi
P568 The effects of red macro algae from Northen coasts of Persian Gulf on melanogenesis
Foroogh Namjoyan, Mojtaba Alishahi, Alireza Jahangiri, Massoumeh Farasat, Hamideh Mousavi
P569 Potential antimicrobial activities of seagrasses, Zostera spp. from Aegean Sea against Vibrio
alginolyticus ATCC 17749
Hijran Yavuzcan, Müjde Eryılmaz, Özlem Bahadır Acıkara, Serkan Özbilgin, Burçin Ergene, Sertel Seçer,
Gülçin Saltan İşcan
P570 Keikipukalides: Furanocembranoid aldehyde derivatives produced by the Antarctic deep sea
soft coral Plumarella delicatissima
Jacqueline L. von Salm, Shane Clark, Prasanth Nemani, Steve Ferlita, Nerida G. Wilson, Bill J. Baker
P571 Protective effect of Oyster hydrolysate peptide in alcohol induced alcoholic fatty liver in SD-‐rats
Jae-‐Hyuk Byun, Yeung-‐Joon Choi, Se-‐Young Choung
P572 FITC-‐labeled dieckol acting as endoplasmic reticulum stress signaling inhibitor in RAW 264.7
macrophage
Jong Hwan Kwak, Jwa-‐Jin Kim, Kyung Bok Lee, Yung Choon Yoo, Jong Seung Kim
P573 Steroids and cembranoids from the soft corals Nephthea erecta and Lobophytum crissum
Jui-‐Hsin Su, Bo-‐Rong Peng, Shih-‐Kai Chou, Ming-‐Cheng Shih, Yu-‐Ting Huang
P574 Bioactive chemical constituents from the Formosan red algae Laurencia tristicha
Jia-‐Yu Chen, Chiung-‐Yao Huang , Jyh-‐Horng Sheu
P575 Structure determination and biosynthesis of the antifungal butyrolactols from Streptomyces sp.
Keebeom Ko, Hui-‐Ming Ge, Jongheon Shin, Dong-‐Chan Oh
P576 Stromatolite microbial communities as a source of new bioactive secondary metabolites
Patricia M. Flatt, Caro Damarjanan, Eric Isamonger, Jarmo C.J. Kalinski, Rosemary A. Dorrington, Kerry
L. McPhail
P577 Isolation, structure elucidation, and total synthesis of an N-‐methylated tetrapeptide from a Pan-‐
amanian marine cyanobacterium
Kh. Tanvir Ahmed, Chakicherla Gayathri, Laura Pineda, Carmenza Spadafora, Roberto Gil, William H.
Gerwick, Kevin J. Tidgewell
P578 NMR characterization of complex natural products: Assigning novel, proton-‐deficient alkaloid
scaffolds
Kirk R. Gustafson, Susanna T. S. Chan, Dennis Milanowski
P579 Antileukemic sesterterpenoids from a marine sponge, Luffariella sp.
Kuei-‐Hung Lai, Mei-‐Chin Lu, Fang-‐Rong Chang, Jui-‐Hsin Su, Mohamed El-‐Shazly, Ying-‐Chi Du, Tung-‐Ying
Wu, Yu-‐Ming Hsu, Anders Backlund, Yang-‐Chang Wu
P580 Isolation and identification of new secondary metabolites from the marine sponge Monanchora
unguiculata
Pierre-‐Eric Campos, Emerson Ferreira Queiroz, Laurence Marcourt, Jean-‐Luc Wolfender, Anne-‐Sophie
Sanchez, Bertrand Illien, Ali Al Mourabit, Anne Gauvin-‐Bialecki
P581 Induction of cryptic metabolites from a rare Antarctic psychrophile, Marinobacter sp.
Jeremy Carter, Miller Judge, Lauren McLean, Jill Mikucki, Lesley-‐Ann Giddings
P582 Biological and chemical analysis of actinomycetes associated with marine sponges from Singa-‐
pore
Gary Chi Ying Ding, Lik Tong Tan
P583 Chemical constituents of the soft coral Sinularia gravis from the coast of Madagascar
Marie Pascaline Rahelivao, Margit Gruner, Hans-‐Joachim Knölker
P584 Antibiotic tetramic acids from marine fungal endophytes cultured under epigenetic regulation
Matthew A. Knestrick, Renee Fleeman, Lindsey N. Shaw, Bill J. Baker
P585 Alpha-‐glucosidase and acetylcholinesterase inhibitory activities of phlorotannins isolated from
the brown alga, Ecklonia maxima
Mutalib
A.
Aderogba,
Rengasamy
R.R.
Kannan,
Ashwell
R.
Ndhlala,
Johannes
Van
Staden
P586
Agelamadins,
bromopyrrole
alkaloids
from
Okinawan
marine
sponges
Agelas
spp.
Naonobu
Tanaka,
Taishi
Kusama,
Yoshiki
Kashiwada,
Jun’ichi
Kobayashi
P587
Bromotyrosine
alkaloids
with
acetylcholinesterase
inhibitory
activity
from
the
Thai
sponge
Acanthodendrilla
sp.
Natchanun
Sirimangkalakitti,
Masashi
Yokoya,
Kanokwan
Changwichit,
Tongchai
Saesong,
Opeyemi
J.
Olatunji,
Supakarn
Chamni,
Pithi
Chanvorachote,
Kornkanok
Ingkaninan,
Anuchit
Plubrukarn,
Naoki
Saito,
Khanit
Suwanborirux
P588
Tools
and
strategies
to
access
to
original
bioactive
compounds
from
cultivation
of
marine
inver-‐
tebrates
and
associated
symbionts:
The
case
of
TASCMAR
Nikolas
Fokialakis,
Ioannis
Trougakos,
Yehuda
Benayahu,
Suchana
Chavanich,
Anne
Bialecki,
Michael
Schaeffer,
Stefano
Zucchinali,
Doru
Felezeu,
Konstantinos
Gardikis,
Pedro
Álvarez,
Åke
Lignell,
Antonel-‐
la
Passani,
Asaf
Ariel,
Jamal
Ouazzani
P589
Marine
halogenated
compound
analysis:
from
an
R
package
to
the
isolation
of
new
griseophe-‐
none
derivatives
Catherine
Roullier,
Yann
Guitton,
Soizic
Prado,
Olivier
Grovel,
Yves
François
Pouchus
P590
Fighting
antibiotic
resistance:
Resensitizing
agents
from
marine
derived
fungi
Paul
Barac,
Henrik
Harms,
Ina
Engels,
Stefan
Kehraus,
Tanja
Schneider,
Gabriele
M.
König
P591
Aceropterin
A,
a
gersolane
diterpene
from
the
Carribean
sea
plume
Pseudopterogorgia
acerosa
(Pallas)
(Gorgonacea)
Paul
Scesa,
Lyndon
West
P592
New
marine
sterols
from
a
Formosan
gorgonian
coral
Pinnigorgia
sp.
Yu-‐Chia
Chang,
Chan-‐Shing
Lin,
Jyh-‐Horng
Sheu,
Ping-‐Jyun
Sung
P593
Anti-‐microbial
metabolites
from
a
marine
bacterium
YMA4
Pi-‐Yu
Chen,
Ning
Lu,
Ying-‐Mi
Lai,
Yu-‐Lang
Yang
P594
Bioactivity
guided
isolation
of
secondary
metabolites
from
a
red
green
Hawaiian
sponge
Ram
P.
Neupane,
Stephen
Parrish,
Wesley
Yoshida,
John
Head,
Richard
Yip,
James
Turkson,
Philip
Wil-‐
liams
P595
Effect
of
alkaloids
isolated
from
Haliclona
sp.
against
hydrogen
peroxide-‐induced
injury
in
SH-‐
SY5Y
human
neuroblastoma
cells
Rebeca
Alvariño,
Eva
Alonso,
Marie-‐Aude
Tribalat,
Olivier
P.
Thomas,
Luis
M.
Botana
P596
Latrunculid
sponges,
their
microbial
communities
and
secondary
metabolites:
connecting
con-‐
served
bacterial
symbionts
to
pyrroloiminoquinone
production.
Rosemary
A
Dorrington,
Storm
H
Hilliar,
Jarmo
CJ
Kalinski,
Rui
WM
Krause,
Kerry
L
McPhail,
Shirley
Parker-‐Nance,
Tara
A
Wlamsley,
Samantha
C
Waterworth
P597
Defensive
chemistry
of
the
Irish
nudibranch
Archidoris
psuedoargus
(Gastropoda
opisthobran-‐
chia)
Ryan
M.
Young,
Bill
J.
Baker
P598
Serratia
marcescens
metabolites
are
promising
candidates
for
biocontrol
of
avocado
pathogens
S.
David
Granada,
Juan
C.
Bedoya-‐Pérez
,
Kirstin
Scherlach,
Christian
Hertweck
P599
Potential
Antimicrobial
Activities
of
Seagrasses,
Zostera
spp.
from
Aegean
Sea;
West
Coast
of
Turkey
Gülçin
Saltan
İşcan,
Özlem
Bahadır
Acıkara,
Müjde
Eryılmaz,
Burçin
Ergene,
Serkan
Özbilgin,
Sertel
Seçer,
Hijran
Yavuzcan
P600
New
diterpene
isolated
from
a
sponge
of
genus
Strongylophora
Stephen
M.
Parrish,
Wesley
Y.
Yoshida,
Philip
G.
Williams
P601
New
polyketides
from
dinoflagellate
Amphidinium
sp.
Takaaki
Kubota,
Takahiro
Iwai,
Hayato
Sato,
Jun’ichi
Kobayashi
P602
Investigation
of
optimal
methods
including
pH-‐zone-‐refining
centrifugal
partition
chromatography
for
the
isolation
of
communesins
from
cultures
of
a
marine-‐derived
Penicillium
expansum
Thuy
T.
P.
Hoang,
Nicolas
Borie,
Audrey
Gratia,
Yves
François
Pouchus,
Catherine
Roullier,
Jean-‐Hugues
Renault,
Olivier
Grovel
P603
Cytotoxic
activity
screening
of
some
Turkish
marine
sponge
species
V.
Murat
Kutluay,
Belma
Konuklugil,
Iclal
Saracoglu
P604
Isolation
and
structure
elucidation
of
new
alkaloids
from
the
bryozoan
Flustra
foliacea
Xiaxia
Di,
Shuqi
Wang,
Eydis
Einarsdottir,
Elin
S.
Olafsdottir,
Sesselja
Omarsdottir
P605
Essential
oil
compostion
and
anticandidal
activity
of
Hypericum
elongatum
L.
subsp.
elongatum
Yavuz
Bülent
Köse,
Gökalp
İşcan,
Mine
Kürkçüoğlu,
Sevim
Küçük
P606
Anti-‐diabetic
and
anti-‐inflammatory
activities
of
the
edible
red
alga
Gracilariopsis
chorda
Yukyoung
Jeon,
Youn
Hee
Nam,
Tong
Ho
Kang,
Jong
Hwan
Kwak
P607
Halogenated
alkaloids
from
Taiwanese
zoanthid
Zoanthus
kuroshio
Yu-‐Ming
Hsu,
I-‐Wen
Lo,
Yang-‐Chang
Wu,
Fang-‐Rong
Chang,
Yuan-‐Bing
Cheng
P608
Two
new
polyhydroxylated
compounds
from
the
fungus
Malbranchea
circinata
with
α
-‐
glucosidase
inhibitory
properties
Abraham
Madariaga-‐Mazón,
Fernando
Cedillo-‐Torres,
Laura
Flores-‐Bocanegra,
Rachel
Mata
P609
Protective
effect
of
oyster
hydrolysate
peptide
in
alcohol
induced
alcoholic
fatty
liver
in
SD-‐rats.
Jae-‐Hyuk
Byun,
Yeung-‐Joon
Choi,
Se-‐Young
Choung
P610
Halogenated
metabolites
from
Irish
Osmundea
spp.
–
a
three
course
meal
for
Aplysia
sp.
Sylvia
Soldatou,
Ryan
M.
Young,
Svenja
Heesch,
Bill
J.
Baker
P611
α-‐Glucosidase
inhibitors
from
Malbranchea
flavorosea
Abraham
Madariaga-‐Mazón,
Daniela
Rebollar-‐Ramos,
Brisa
Verastegui-‐Omaña,
Laura
Flores-‐
Bocanegra,
Rachel
Mata
P612
Statistical
optimization
of
production
and
isolation
of
cytotoxic
compound
from
Nocardia
igno-‐
rata
Alba
Noël,
Gwendoline
Van
Soen,
Isabelle
Rouaud,
Solenn
Ferron,
Eric
Hitti,
Sophie
Tomasi
P613
Characterization
of
alkaloids
and
carotenoids,
a
defense
cocktail
on
Coccinellidae
eggs’
surface.
Alexandre
Maciuk,
Pedro
Vásquez-‐Ocmín,
Felipe
Ramon-‐Portugal,
Gilles
Espinasse,
Erwan
Poupon,
Alexandra
Magro
P614
Sectorial
land
snail
damage
to
the
lichen
Argopsis
friesiana
could
be
explained
by
metabolite
profiles.
Alice
Gadéa,
Françoise
Le
Dévéhat,
Anne-‐Cécile
Le
Lamer,
Pierre
Le
Pogam,
Damien
Ertz,
Maryvonne
Charrier,
Joël
Boustie
P615
SOD
inhibitors
research
from
endophytics
fungi
extracts
Alix
Poinso,
Patricia
Vicendo,
François
Couderc,
Nicolas
Fabre,
Guillaume
Marti,
Marieke
Vansteelandt,
Mohamed
Haddad,
Billy
J.
Cabanillas,
Varravaddheay
Ong-‐Meang
P616
Bacterial
identification
through
machine
learning
Andreas
Helfenstein,
Päivi
Tammela
P617
Isolation
and
characterization
of
halogenated
monoterpenes
in
the
investigation
of
the
ecologi-‐
cal
relationship
between
Antarctic
Plocamium
cartilagineum
and
Paradexamine
fissicauda
Andrew
J.
Shilling,
Jacqueline
L.
von
Salm,
Ryan
M.
Young,
Margaret
O.
Amsler,
Charles
D.
Amsler,
James
B.
McClintock,
Bill
J.
Baker
P618
Trichaptum
abietinum
from
British
Columbia
exhibited
anti-‐proliferative
and
immuno-‐
modulatory
activities
Ankush
Barad,
Sumreen
Javed,
Chow
H.
Lee
P619
Isolation
of
metabolites
from
epigenetically
modified
mangrove
fungi
for
anti-‐infective
drug
discovery
Anne-‐Claire
D.
Limon,
Renee
Fleeman,
Lindsey
N.
Shaw,
Bill
J.
Baker
P620
The
new
diketopiperazines
produced
by
marine
algicolous
fungus
Penicillium
sp.
KMM
4672
Anton
N.
Yurchenko,
Olga
F.
Smetanina
P621
De
novo
metabolite
production
through
co-‐cultivation
of
different
fungal
species
on
solid
media
Antonio
Azzollini,
Jean-‐Luc
Wolfender,
Katia
Gindro
P622
Cytotoxic
activity
of
endolichenic
fungi
isolated
from
the
lichen
Nephroma
laevigatum.
Aurélie
Lagarde,
Marion
Millot,
Patricia
Jargeat,
Tan-‐Sothéa
Ouk,
Vincent
Sol,
Lengo
Mambu
P623
New
bioactive
compounds
from
Fusarium
fujikuroi
Birgit
Arndt,
Slavica
Janevska,
Isabel
Krug,
Lucas
Maciel
Mauriz
Marques,
Bettina
Tudzynski,
Hans-‐
Ulrich
Humpf
P624
Influence
of
Inonotus
hispidus
on
function
of
human
immune
cells
Carsten
Gründemann,
Mandy
Arnhold,
Stefanie
Meier,
Christian
Bäcker,
Manuel
Garcia-‐Käufer,
Fran-‐
ziska
Grunewald,
Roman
Huber,
Ulrike
Lindequist
P625
Anticancer
active
metabolites
from
soil
bacteria
Cassandra
Lew,
Chenxi
Zoe,
Sandra
Loesgen
P626
The
discovery
and
evaluation
of
anti-‐parasitic
metabolites
from
filamentous
fungi
Cedric
Pearce,
Blaise
Darveaux,
Christopher
Rice,
Beatrice
Colon,
Kaitlin
Mettel,
Kati
Rasanen,
Dennis
Kyle,
Bill
Baker,
Nicholas
Oberlies
P627
A
UHPLC/MS-‐MS-‐based
HDAC
assay
applied
to
bio-‐guided
microfractionation
of
fungi
extracts
Vincent
Zwick,
Claudia
A.
Simões-‐Pires,
Lucie
Ory,
Pierre-‐Marie
Allard,
Jean-‐Luc
Wolfender,
Muriel
Cuendet
P628
A
cytotoxic
pentadecapeptide
from
a
South
African
Didemnid
tunicate
David
Gallegos,
Jeffrey
Serrill,
Shirley
Parker-‐Nance,
Rosemary
Dorrington,
Jane
Ishmael,
Kerry
McPhail
P629
Insect
symbionts:
Prolific
sources
of
new
bioactive
compounds
Dong-‐Chan
Oh
P630
Characterisation
of
vietnamycin:
a
novel
Burkholderia
antibiotic
targeting
mupirocin-‐resistant
methicillin-‐resistant
Staphylococcus
aureus
(MRSA)
Rachel
A.
Rowe,
Cerith
Jones,
Matthew
J.
Bull,
Matthew
Jenner,
Lijang
Song,
Yousef
Dashti,
Simon
R.
Harris,
Julian
Parkhill,
Thomas
R.
Connor,
Gregory
L.
Challis,
Eshwar
Mahenthiralingam
P631
New
antiplasmodial
compounds
discovered
by
High
Throughput
Screening
(HTS)
of
a
collection
of
microbial
natural
extracts
Noureddine
El
Aouad,
Frederick
Annang,
Ignacio
Pérez-‐Victoria,
Jesús
Martín,
Gloria
Crespo,
Elizabeth
Domingo,
Guiomar
Pérez-‐Moreno,
Juan
Cantizani,
Paula
Sánchez-‐Carrasco,
Ignacio
González,
Víctor
González-‐Menéndez,
Nuria
de
Pedro,
José
R.
Tormo,
Luis
M.
Ruiz-‐Pérez,
Dolores
González-‐Pacanowska,
Francisca
Vicente,
Gerald
F.
Bills,
Olga
Genilloud,
Fernando
Reyes
P632
Decalin-‐containing
polyketides
with
antibacterial
activities
from
an
endophytic
fungus,
Eupeni-‐
cillium
sp.
LG41
Gang
Li,
Souvik
Kusari,
Hartmut
Laatsch,
Christopher
Golz,
Carsten
Strohmann,
Michael
Spiteller
P632a
An
endophytic
fungus
Phyllosticta
capitalensis
harboring
uncultivable
endosymbiotic
bacterium
Herbaspirillum
sp.
produces
lactam-‐fused
4-‐pyrones
Wen-‐Xuan
Wang,
Souvik
Kusari,
Parijat
Kusari,
Oliver
Kayser,
Michael
Spiteller
P633
Two
new
compounds
produced
by
Xylaria
sp.
an
endophytic
fungus
from
Casearia
sylvestris
(Flacourtiaceae)
G.
F.
Martins,
C.
R.
Nogueira,
G.
H.
Silva,
M.
C.
M.
Young,
C.
O.
Pessoa,
D.
J.
B.
Lima,
P.
M.
P.
Ferreira,
V.
S.
Bolzani,
A.
R.
Araujo
P634
Antifungal
long-‐chain
alkenyl
sulphates
isolated
from
culture
broths
of
the
fungus
Chaetopsina
sp.
Gloria
Crespo,
Ignacio
Pérez
Victoria,
Mercedes
de
la
Cruz,
Víctor
González-‐Menéndez,
Bastien
Cautain,
Jesús
Martín,
Francisca
Vicente,
Olga
Genilloud,
Fernando
Reyes
P635
Fungi
as
a
source
for
antibacterial
compounds
Jawad
Anwar,
Taj
Muhammad,
Ulf
Göransson,
Zafar
Iqbal
P636
Production
and
detection
of
the
natural
ionophore
Beauvericin
Jesús
M.
González,
Amparo
Alfonso,
MJ
Sainz
,
Luis
M
Botana
P637
Peptaibols
from
Tichoderma
sp.
(MSX70741):
Isolation,
structure
elucidation
and
biological
activity
José
Rivera-‐Chávez,
Huzefa
A.
Raja,
Prashant
Metri,
Ding
Xue,
Cedric
J.
Pearce,
Nicholas
H.
Oberlies
P638
Development
of
microcystin
derivatives
as
novel
agents
against
OATP1B3-‐expressing
tumors
Julius
Krivec,
Urs
F.
Moschik,
Wolfram
Lorenzen,
Julia
Moschny,
Timo
H.
J.
Niedermeyer
P639
Bioguided
isolation
as
a
tool
for
the
discovery
of
novel
cosmeuceutical
agents
from
microbial
biodiversity
Katerina
Georgousaki,
Nikolaos
Tsafantakis,
Sentiljana
Gumeni,
Spiros
Fotinos,
Ioannis
P.
Trougakos,
Nikolas
Fokialakis
P640
Do
the
secondary
metabolites
from
Phanerochaete
chrysosporium
change
depending
on
their
growth
substrates?
Kirk
P.
Manfredi,
James
Humpal
P641
Chemotaxonomy
of
the
genus
Stemphylium
Kresten
Jon
Kromphardt
Olsen,
Birgitte
Andersen
P642
Cytotoxic
activities
of
endophytic
fungi
extracts
from
Paepalanthus
planifolius:
A
Brazilian
evergreen
Marcelo
R.
de
Amorim,
Weslei
B.
Botero,
Ana
Caroline
Z.
Silva,
Angela
R.
Araújo,
Iracilda
Z.
Carlos,
Lourdes
Campaner
dos
Santos
P643
Influence
of
the
medium
and
preferred
cereal
substrate
on
secondary
metabolite
production
by
species
from
Penicillium
series
Viridicata
Magnus
Hallas-‐Møller,
Kristian
F.
Nielsen
and
Jens
C.
Frisvad
P644
New
diketomorpholines
from
a
facultative
marine-‐derived
Aspergillus
sp.
(ACA-‐9)
Manuel
A.
Aparicio-‐Cuevas,
Isabel
Rivero-‐Cruz,
María
C.
González,
Huzefa
A.
Raja,
Mario
Figueroa
P645
Evaluation
of
lichen
compounds
as
inhibitors
of
Candida
albicans
biofilms
Marion
Girardot,
Marion
Millot,
Clément
Bernard,
Lengo
Mambu,
Christine
Imbert
P646
Screening
of
actinobacteria
for
inhibition
of
quorum
sensing
of
Chromobacterium
violaceum
CV026
and
Staphylococcus
aureus
PC322
Nico
Ortlieb,
Timo
H.
J.
Niedermeyer
P647
Bioactive
type
A
proanthocyanins
from
fungus
Laurobasidium
lauri
João
Serina,
Maria
J.
Carvalho,
Tatiana
Weinhold,
Paula
C.
Castilho
P648
Modulation
of
a
lichen-‐associated
fungus
for
improved
biosynthesis
of
bioactive
secondary
me-‐
tabolites
Peter
M.
Eze,
Blessing
O.
Umeokoli,
Huiqin
Chen,
Zhen
Liu,
Festus
B.C.
Okoye,
Charles
O.
Esimone,
Peter
Proksch
P649
Screening
of
cyanobacteria
extracts
for
inhibitory
activity
against
the
cysteine
protease
rhodesain
of
Trypanosoma
brucei
Ronja
Kossack,
Trang
Nguyen,
Steffen
Breinlinger,
Tanja
Schirmeister,
Timo
H.
J.
Niedermeyer
P650
Mensacarcin,
a
soil
derived
polyketide,
exhibits
strong
universal
anti-‐proliferative
effects
in
human
cancer
cell
lines
and
induces
cell
death
selectively
in
melanoma
cells.
Birte
Plitzko,
Sandra
Loesgen
P651
Chemical
investigations
of
the
fruiting
bodies
of
Pleurotus
cornucopiae
and
biological
activities
of
the
isolated
compounds
Seoung
Rak
Lee,
Seulah
Lee,
Hee
Jeong
Eom,
Hee
Rae
Kang,
Jae
Sik
Yu,
Tae
Kyoung
Lee,
Jiwon
Baek,
Dahae
Lee,
Won
Se
Suh,
Ki
Hyun
Kim
P652
Inhibitory
effect
of
isolated
constituents
from
sclerotia
of
Poria
cocos
on
LPS-‐induced
NO
pro-‐
duction
Seoung
Rak
Lee,
Seulah
Lee,
Hee
Jeong
Eom,
Hee
Rae
Kang,
Jae
Sik
Yu,
Tae
Kyoung
Lee,
Jiwon
Baek,
Dahae
Lee,
Won
Se
Suh,
Ki
Hyun
Kim
P653
Marine
fungi:
A
novel
source
of
cosmeceutical
applications
Shivankar
Agrawal,
Sunil
Kumar
Deshmukh,
Colin
Barrow,
Alok
Adholeya
P654
Indole
alkaloids
from
Penicillium
genus:
Identification,
isolation,
structure
elucidation
and
cyto-‐
toxicity
Svetlana
A
Kalinina,
Annika
Jagels,
Benedikt
Cramer,
Hans-‐Ulrich
Humpf
P655
Activation
of
fungal
secondary
metabolism
by
disturbance
of
the
two-‐component
signal
trans-‐
duction
system
and
identification
of
the
nectriapyrone
biosynthetic
gene
cluster
Takayuki
Motoyama,
Yoko
Tanaka,
Hiroyuki
Osada
P656
Prediction
of
secondary
metabolite
encoding
genes
based
on
chemical
structure
analysis
Thomas
Isbrandt,
Maria
Lund
Nielsen,
Casper
Hoeck,
Yuksel
Gezgin,
Rasmus
J.
N.
Frandsen,
Kristian
Fog
Nielsen,
Thomas
Ostenfeld
Larsen
P657
Chemical
and
biological
investigation
of
Algerian
lichens
Rafika
Brakni,
Monia
Serradj,
Xavier
Fernandez,
Thomas
Michel
P658
Insects-‐entomopathogens
interactions
to
discover
new
insecticidal
and
antimicrobial
com-‐
pounds
Seinde
Touré,
I.
Dusfour,
D.
Stien,
V.
Eparvier
P659
New
antibacterial
small
molecules
from
insect-‐associated
bacteria
in
Bombyx
mori
and
Nicrophorus
concolor
Yern-‐Hyerk
Shin,
Ki-‐Bong
Oh,
Jongheon
Shin,
Dong-‐Chan
Oh
P660
Application
of
a
simple
bioactivity
profiling
strategy
to
natural
product
discovery
from
endo-‐
phytes
of
marine
macroalgae
Andrew
J.
Flewelling,
John
A.
Johnson,
Christopher
A.
Gray
P661
Rearranged
sesquiterpenes
produced
by
Camarops
sp.
an
endophytic
fungus
in
Alibertia
macro-‐
phylla
(Rubiaceae)
Juliana
R.
Gubiani,
Cláudio
R.
Nogueira,
Maria
C.
M.
Young,
Paulo
M.
P.
Ferreira,
Manoel
O.
de
Moraes,
Cláudia
Pessoa,
Vanderlan
S.
Bolzani,
Angela
R.
Araujo
P662
Search
for
novel
metabolites
in
fungal
endophytes:
study
of
Phomopsis
sp.
and
Colletotrichum
sp.
co-‐cultivation
and
Botryosphaeria
mamane
epigenetic
modification
Asih
Triastuti,
Marieke
Vansteelandt,
Fatima
Barakat,
Patricia
Jargeat,
Laura
Rieusset,
Nicolas
Fabre,
Carlos
Amasifuen,
Alexis
Valentin,
Mohamed
Haddad
P663
Differential
chemotypes
produce
phenotypic
responses
in
endophytic
microbial
interactions
Caraballo-‐Rodriguez,
A.
M.,
Pupo,
M.
T.
P664
Effect
of
germination
on
in
vitro
and
in
vivo
activity
of
Lupinus
albus
L.
and
Lupinus
angustifolius
L.
seed
extract
Corina
Danciu,
Ersilia
Alexa,
Istvan
Zupko,
Stefana
Avram,
Ioana
Zinuca
Pavel,
Daliana
Minda,
Georgeta
Pop,
Dorina
Coricovac,
Cristina
Dehelean
P665
Co-‐cultivation
approach
and
untargeted
metabolomics
in
the
search
for
new
secondary
metabo-‐
lites
from
endophytic
fungi
Fatima
Barakat,
Marieke
Vansteelandt,
Asih
Triastuti,
Laura
Rieusset,
Billy
Cabanillas,
Mohamed
Had-‐
dad,
Nicolas
Fabre
P666
Proteolytic
active
proteins
from
Euphorbia
mauritanica
L.
stimulate
the
production
of
interleu-‐
kine-‐6
and
interleukine-‐8
in
keratinocytic
HaCaT
cells
Florian
Guenther,
Matthias
F.
Melzig
P667
Antigenotoxic
compounds
from
bark
of
South
African
Erythrina
latissima
Yancho
Zarev,
Kenn
Foubert,
Vera
Almeida,
Sandra
Apers,
Luc
Verschaeve,
Luc
Pieters
P668
Chemical
constituents
and
biological
activity
from
the
stem
and
root
of
Neolitsea
konishii
Hsien-‐Kai
Huang,
Shan-‐Yu
Lin,
Su-‐Ling
Wong,
Tian-‐Lu
Cheng,
Chu-‐Hung
Lin,
Kim-‐Hong
Gan,
Hsun-‐Shuo
Chang,
Ih-‐Sheng
Chen
P669
Cytotoxic
lignans
from
the
fruits
of
Koelreuteria
henryi
Chu-‐Hung
Lin,
Kuo-‐Hsiung
Lee,
Ih-‐Sheng
Chen,
Hsun-‐Shuo
Chang
P670
Three
new
dikeopiperazines
from
Costa
Rican
endolichenic
fungus
Colpoma
sp.
CR1465A
Jae
Sik
Yu,
Seulah
Lee,
Hee
Jeong
Eom,
Hee
Rae
Kang,
Seoung
Rak
Lee,
Tae
Kyoung
Lee,
Jiwon
Baek,
Dahae
Lee,
Won
Se
Suh,
Ki
Hyun
Kim
P671
Resorcylic
lactones
from
Lasiodiplodia
theobromae
(MUB65),
a
fungal
endophyte
isolated
from
Myracrodruon
urundeuva
Aline
C.
M.
Sobreira,
Otília
D.
L.
Pessoa,
Katharine
G.
D.
Florêncio,
Diego
V.
Wilke,
Francisco
C.
O.
Freire,
Francisco
J.
T.
Gonçalves,
Paulo
Riceli.
V.
Ribeiro,
Lorena
M.
A.
Silva,
Edy.
S.
Brito,
Kirley
M.
Canuto
P672
Biodiversity
of
fungal
community
associated
to
lichens
active
against
Candida
biofilms
Patricia
Jargeat,
Marion
Girardot,
Caroline
Rouger,
Willy
Aucher,
Marion
Millot,
Christine
Imbert,
Lengo
Mambu
P673
Increasing
the
production
of
secondary
metabolites
by
changes
in
culture
conditions
Madelinea
Aguilar,
Maria
Julca-‐Canto,
Nivia
Rios,
Luis
Cubilla-‐Rios
P674
New
hormonemate
derivatives
from
the
endophytic
fungus
Dothiora
sp.
Mercedes
Pérez-‐Bonilla,
Víctor
González-‐Menendez,
Nuria
de
Pedro,
Ignacio
Pérez-‐Victoria,
Jesús
Mar-‐
tín,
Francisca
Vicente,
Olga
Genilloud,
José
R.
Tormo,
Fernando
Reyes
P675
Angiogenesis
inhibitors
and
anti-‐inflammatory
agents
from
Phoma
sp.
NTOU4195
Ming-‐Shain
Lee,
Shin-‐Wei
Wang,
Guei-‐Jane
Wang,
Ka-‐Lai
Pang,
Ching-‐Kuo
Lee,
Yueh-‐Hsiung
Kuo,
Hyo-‐
Jung
Cha,
Ruo-‐Kai
Lin,
Tzong-‐Huei
Lee
P676
Discovery
of
new
bioactive
secondary
metabolites
produced
by
Streptomyces
strains
derived
from
volcanic
islands
Munhyung
Bae,
Heegyu
Kim,
Sohyun
Park,
Jongheon
Shin,
Ki-‐Bong
Oh,
Sang
Kook
Lee,
Dong-‐Chan
Oh
P677
Biocontrol
potential
of
endophytic
fungi
against
Batrachochytrium
dendrobatidis,
the
fungi
causing
global
amphibian
declines
Carolina
Castro,
Carolina
Portero,
Alexandra
Narváez-‐Trujillo
P678
Chemical
study
on
Paenibacillus
odorifer,
a
bacterial
species
isolated
from
lichen
Nguyen
Thi
Bach
Le,
Delmail
David,
Tomasi
Sophie
P679
Bigger
is
not
always
better:
A
study
of
the
structure-‐activity
relationship
of
oligomeric
ellag-‐
itannins
on
ruminal
fermentation
in
vitro
Nicolas
Baert,
Wilbert
F.
Pellikaan,
Maarit
Karonen,
Juha-‐Pekka
Salminen
P680
Elicitation
of
isoflavonoid
production
in
the
cell
suspension
culture
of
Pueraria
candollei
var.
candollei
by
endophytic
bacteria
Panitch
[Boonsnongcheep],
Atsuko
Matsumoto,
Yoko
Takahashi,
Sompop
Prathanturarug
P681
A
new
indole
alkaloid
from
the
endophyte
Aspergillus
ustus
isolated
from
the
mangrove,
Avi-‐
cennia
germinans
Petrea
C.
Facey,
Roy
B.
Porter,
Hartmut
Laatsch
P682
Compounds
with
anti-‐respiratory
syncytial
virus
activity
from
endophytic
fungus
Pestalotiopsis
thea
Philip
F.
Uzor1,
Damian
C.
Odimegwu,
Weaam
Ebrahim,
Patience
O.
Osadebe,
Ngozi
J.
Nwodo,
Fesus
B.
C.
Okoye,
Zhen
Liu,
Peter
Proksch
P683
The
endophyte
Candidatus
Burkholderia
crenata
of
the
TCM
plant
Ardisia
crenata
produces
the
selective
Gq-‐inhibitor
FR900359
Raphael
Reher,
Isabella
Schamari,
Stefan
Kehraus,
Suvi
Annala,
Markus
Kuschak,
Till
Schäberle,
Max
Crüsemann,
Aurelien
Carlier,
Leo
Eberl,
Christa
E.
Müller,
Evi
Kostenis,
Gabriele
M.
König
P684
Isolation
of
bioactive
secondary
metabolites
from
mangrove
fungal
endophytes
using
epigenetic
regulation
Santana
A.
L.
Thomas,
Renee
Fleming,
Lindsey
N.
Shaw,
Bill
J.
Baker
P685
Secondary
metabolites
from
the
fungus
Porodaedalea
pini
Shuen-‐Shin
Yang,
Hing-‐Yuen
Chan,
Sung-‐Yuan
Hsieh,
Gwo-‐Fang
Yuan,
Su-‐Ling
Wong,
Chu-‐Hung
Lin,
Ming-‐Jen
Cheng,
Ih-‐Sheng
Chen,
Hsun-‐Shuo
Chang
P686
Development
of
chemopreventive
agents
for
hepatocellular
carcinoma
from
Excoecaria
for-‐
mosana
by
a
glycine
N-‐methyltransferase
(GNMT)
gene
expression-‐oriented
screen
platform
Ho-‐Cheng
Wu,
Chia-‐Hung
Yen,
Yi-‐Ming
Chen,
Ya-‐Han
Chang,
Hsun-‐Shuo
Chang
P687
Mangrove
associated
fungi
as
a
source
of
potential
drugs
against
the
ESKAPE
pathogens
Sofia
Kokkaliari,
Renee
Fleeman,
Lindsey
N.
Shaw,
Bill
J.
Baker
P688
New
bioactive
secondary
metabolites
from
dung
beetle-‐associated
bacteria
Soohyun
Um,
Ki-‐Bong
Oh,
Jongheon
Shin,
Dong-‐Chan
Oh
P689
Novel
insights
into
plant-‐endophyte
communication:
maytansine
as
an
example
Souvik
Kusari,
Parijat
Kusari,
Dennis
Eckelmann,
Sebastian
Zühlke,
Oliver
Kayser,
Michael
Spiteller
P690
A
comparison
between
the
application
of
NMR
and
LC-‐HRMS
based
metabolomics
on
the
discov-‐
ery
of
natural
products
from
endophytic
fungi
Trevor
N.
Clark,
Fabrice
Berrué,
Larry
Calhoun,
Patricia
Boland,
Russel
Kerr,
John
A.
Johnson,
Christo-‐
pher
A.
Gray
P691
Unusual
natural
products
mediated
root
hairs-‐endophyte
stacking
(RHESt)
that
traps
and
kills
pathogens
Walaa
K.
Mousa,
Charles
Shearer,
Cassandra
Ettinger,
Jonathan
Eisen,
Manish
N.
Raizada
P693
Search
for
anti-‐dengue
secondary
metabolites
from
two
Diospyros
endophytes,
Aspergillus
sp.
and
Phomopsis
sp.
Laure-‐Anne
Peyrat,
Véronique
Eparvier,
Cécilia
Eydoux,
Jean-‐Claude
Guillemot,
Didier
Stien,
Marc
Litaudon
P694
Anti-‐MRSA
natural
products
from
an
epigenetic
modified
Floridian
mangrove-‐associated
fungus
Sylvia
Soldatou,
Renee
Fleeman,
Lindsey
N.
Shaw,
Bill
J.
Baker
P695
Bioinspired
total
syntheses
of
(±)-‐dictazole
B
and
(±)-‐tubastrindole
B:
"The
aplysinopsins’
cas-‐
cade"
Adam
Skiredj,
Mehdi
A.
Beniddir,
Delphine
Joseph,
Guillaume
Bernadat,
Laurent
Evanno,
Erwan
Pou-‐
pon
P696
Harnessing
the
main
event
of
drimentines
biosynthesis:
bio-‐inspired
synthesis
and
biological
evaluation
of
Δ8’-‐isodrimentine
A
and
related
compounds
Adam
Skiredj,
Mehdi
A.
Beniddir,
Laurent
Evanno,
Erwan
Poupon
P697
Enzymatic
tailoring
of
oleuropein
isolated
from
Olea
europaea
leaves
Efstratios
Nikolaivits,
Aikaterini
Termentzi,
Alexios-‐Leandros
Skaltsounis,
Nikolas
Fokialakis,
Evange-‐
los
Topakas
P698
HPLC
quantification
method
for
chrysin
and
tectochrysin
in
Flourensia
extracts
M.
Ángeles
Ramírez-‐Cisneros,
Ramiro
Ríos
Gómez,
María
Yolanda
Rios
P699
Synthesis
of
cyclic
citrullinated
peptides
targeting
rheumatoid
arthritis
autoantibodies
based
on
sunflower
trypsin
inhibitors
Camilla
Eriksson,
Sunithi
Gunasekera,
Cátia
Cerqueira,
Per-‐Johan
Jacobsson,
Ulf
Göransson
P700
Silymarin
from
Silybum
marianum
–
new
approaches
to
separation
and
derivatization
David
Biedermann,
Alena
Křenková,
Kateřina
Valentová,
Vladimír
Křen
P701
Boosting
the
antifungal
drug
discovery
by
halogenating
plant
extracts
to
obtain
bioactive
‘un-‐
natural’
natural
products
Davide
Righi,
Alice
Mainetti,
Quentin-‐Favre
Godal,
Laurence
Marcourt,
Jean-‐Luc
Wolfender,
Emerson
F.
Queiroz
P702
Sarqaquinoic
acid
and
related
synthetic
naphthoquinones
inhibit
the
function
of
Hsp90
Maynard
Chiwakata,
Jo-‐Anne
de
la
Mare,
Adrienne
Edkins,
Denzil
R.
Beukes
P703
Assessment
of
the
bactericidal
effect
of
green
synthesized
silver
nanoparticles
against
a
panel
of
infectious
microorganisms
Edith
Antunes,
Mokone
Mmola,
Mervin
Meyer,
Denzil
R.
Beukes
P704
Biomimetic
studies
towards
the
total
synthesis
of
highly
complex
araiosamines
Kévin
Cottet,
Mehdi
A.
Beniddir,
Adam
Skiredj,
Laurent
Evanno,
Erwan
Poupon
P705
Antimalarial
and
antimicrobial
activities
of
α-‐(2-‐piperidyl)-‐2-‐aryl-‐4-‐quinolinemethanol
analogs
H.
M.
T.
Bandara
Herath,
H.
Ranjith
W.
Dharmaratne,
Melissa
Jacob,
Shabana
I.
Khan,
N.
P.
Dhammika
Nanayakkara
P706
Establishment
and
evaluation
of
processes
for
the
production
of
the
antiviral
and
cytostatic
cardenolide
glucoevatromonoside
Jennifer
Munkert,
Marina
Santiago
Franco,
Fernao
C.
Braga,
Wolfgang
Kreis,
Saulo
F.
Andrade,
Flaviano
Melo
Ottoni,
Ricardo
José
Alves,
Rodrigo
Maia
de
Pádua
P707
Synthesis
and
characterization
99mTc-‐labebled
DTPA-‐digitoxigenin
and
its
new
potential
in
im-‐
aging
techniques
for
the
diagnostic
and
identification
of
tumor
cells
Jennifer
Munkert,
Eliza
Rocha
Gomes,
Saulo
F.
Andrade,
José
Dias
de
Souza
Filho,
Lucas
L.
Marostica,
Wolfgang
Kreis,
Fernão
C.
Braga,
Cláudia
M.
O.
Simões,
Valbert
Nascimento
Cardoso,
Rodrigo
M.
Pádua,
André
Luís
Branco
de
Barros
P708
Four
new
diterpenoid
alkaloids
from
Aconitum
japonicum
Koji
Wada,
Keiko
Takeda,
Machiko
Haraguchi,
Yuki
Abe,
Natsumi
Kuwahara,
Shota
Suzuki,
Ayaka
Terui,
Takumi
Masaka,
Naoko
Munakata,
Mariko
Uchida,
Masashi
Nunokawa,
Hiroshi
Yamashita,
Masuo
Goto,
Kuo-‐Hsiung
Lee
P709
Phytotoxic
constituents
of
Diaporthe
eres
and
synthesis
of
analogs
Kumudini
M.
Meepagala,
Natascha
Techen,
Robert
D.
Johnson,
Stephen
O.
Duke
P710
A
flavone
and
cytotoxic
activity
of
sesquiterpenoids
from
the
resinous
exudates
of
cushion
bush
(Leucophyta
brownii)
Katrine
T.
Jensen,
Mette
G.
Hyldgaard,
Stig
Purup,
Xavier
Fretté,
Lars
P.
Christensen
P711
Biomimetic
assembly
of
leucoridine
A
Sarah
Benayad,
Mehdi
A.
Beniddir,
Laurent
Evanno,
Erwan
Poupon
P712
Preakuammicine:
A
long
awaited
missing
link
in
the
biosynthesis
of
monoterpene
indole
alka-‐
loids
Sarah
Benayad,
Kadiria
Ahamada,
Guy
Lewin,
Laurent
Evanno,
Erwan
Poupon
P713
Cytotoxic
potential
of
naturally
occurring
isoquinoline
alkaloids
possessing
different
structural
types
Cahlíková
Lucie,
Doskočil
Ivo,
Chlebek
Jakub,
Hošťálková
Anna,
Havelek
Radim,
Šafratová
Marcela
P714
Isolation
and
structure
elucidation
of
twelve
new
prenylated
flavonoids
from
Onobrychis
spp.
(Leguminosae)
employing
LC-‐HRMS,
HSCCC
and
NMR
techniques
Maria-‐Eleni
Sakavitsi,
Job
Tchoumtchoua,
Sofia
Mitakou,
Maria
Halabalaki,
Alexios-‐Leandros
Skaltsounis
P715
B-‐ring
modification
in
prenylflavonoids
of
hops
and
the
effect
on
induction
of
neuronal
differen-‐
tiation
Michael
Kirchinger,
Corinna
Urmann,
Lara
Bieler,
Sebastien
Couillard-‐Despres,
Herbert
Riepl
P716
A
peptide
isolated
from
an
ant
active
against
Helicobacter
pylori
J.
Guzman,
M.
Treilhou,
D.
Castillo,
H.
Belkhelfa,
L.
Haddioui-‐Hbabi,
M.
Sauvain
P717
Anatomical
characterization
and
chemical
profiling
of
Rumex
species
Hye-‐Jin
Kim,
Woo
Sung
Park,
Ji-‐Yeong
Bae,
Jong
Hee
Park,
Mi-‐Jeong
Ahn
P718
Volatile
compounds
of
Tripleurospermum
decipiens
from
different
sites
in
Turkey
Mine
Kurkcuoglu,
Fatma
Tosun,
Huseyin
Inceer,
K.
Husnu
Can
Baser
P719
Phytochemical
and
cytotoxic
studies
on
Cestrum
nocturnum
Mona
A.
Mohamed
P720
Bioactive
formylated
flavonoids
from
Eugenia
rigida:
Their
isolation,
synthesis,
and
X-‐ray
crys-‐
tallography
Muhammad
Ilias,
Mohamed
A.
Zaki,
Melissa
R.
Jacob,
Shabana
I.
Khan,
Mohamed
A.
Ibrahim,
Volodymyr
Samoylenko,
Rabab
Mohammed,
Mona
H.
Hetta,
David
D.
Pasco,
Daneel
Ferreira,
Frank
R.
Fronczek,
Dhammika
Nanayakkara
P721
Flavone
derivatives:
a
promising
tool
in
the
fight
against
malaria
Flore
Nardella,
Silvia
Stiebing,
Patrick
Wagner,
Valérie
Collot,
Marcel
Kaiser,
Benoit
Witkowski,
Didier
Menard,
Martine
Schmitt,
Ermanno
Candolfi,
Catherine
Vonthron-‐Sénécheau
P722
A
two-‐season
impact
study
on
Globularia
alypum:
Adaptive
leaf
structures
and
metabolic
varia-‐
tions
Stavroula
Mamoucha,
Nikolaos
Tsafantakis,
Nikolas
Fokialakis,
Nikolaos
Christodoulakis
P723
Synthesis
and
thymidine
phosphorylase
inhibition
studies
of
5-‐chlorobenzothiazole
derivatives
Mastura
Arbin,
Norizan
Ahmat,Muhammad
Taha
P724
Lupeol
and
resveratrol
from
the
stembark
of
Shorea
ovalis
(Dipterocarpaceae)
Rosmawati
A.
Aziz,
N.
Ahmat
P725
Synthesis
of
benzimidazole
derivatives
as
new
α-‐Glucosidase
inhibitors
Nik
Khairunissa
Nik
Abdullah
Zawawi,
Norizan
Ahmat,
Muhammad
Taha,
Aisyah
Salihah
Kamarozaman
P726
New
cassane
diterpenes
from
the
leaves
and
twigs
of
Caesalpinia
bonduc
(Linn)
Roxb
Olubanke
O.
Ogunlana,
Wen
J.
He,
Jun
T.
Fan,
Guang
Z.
Zeng,
Oluseyi
E.
Ogunlana,
Abiodun
H.
Adebayo,
Chang
J.
Ji,
Joseph
O.
Olagunju,
Afolabi
A.
Akindahunsi,
Ning
H.
Tan
P727
Microbial
transformation
of
ruscogenins
by
Cunninghamella
blakesleeana
Özge
Özçinar,
Özgür
Tağ,
Bijen
Kivçak,
Erdal
Bedir
P728
Synthetic
derivatives
of
pulchrol:
An
antiparasitic
natural
product
Paola Terrazas, Olov Sterner
P729 Rational design of annotine analogues with increased inhibitory activity towards acetylcholin-‐
esterase
Elsa S. Halldorsdottir, Sebastian Oddsson, Arndis M. Einarsdottir, Borghildur Eiriksdottir, Natalia M.
Kowal, Elin S. Olafsdottir
P730 (‒)-‐Bassianolide, a cyclodepsipeptide from Bombycis Corpus: Total synthesis and evaluation of
its antitumor activity
Seoung Rak Lee, Seulah Lee, Hee Jeong Eom, Hee Rae Kang, Jae Sik Yu, Tae Kyoung Lee, Jiwon Baek,
Dahae Lee, Won Se Suh, Ki Hyun Kim
P731 The effect of prenylation on the antimicrobial activity of selected naturally occurring furanones
and pyranones
Serena Fiorito, Francesco Epifano, Vito Alessandro Taddeo, Salvatore Genovese, Jabrane Azelmat, Da-‐
niel Grenier
P732 Preliminary investigations on seleno-‐analogues of plant oxyprenylated secondary metabolites
Serena Fiorito, Francesco Epifano, Vito Alessandro Taddeo, Salvatore Genovese, Luca Sancineto, Clau-‐
dio Santi
P733 Chemically engineered extracts of St John’s wort as sources of polyprenylated acylphloroglu-‐
cinols to prevent endothelial dysfunction
Maxime Le Bot, Nina Corlay, Aurore Michaud, Sylvain Pagie, Pascal Richomme, Béatrice Charreau, Sé-‐
verine Derbré
P734 Design, synthesis and biological evaluation of novel emodin derivatives as potent anti-‐
dyslipidemic and antioxidant agents
Sukanya Pandeti, Ravi Sonkar, Gitika Bhatia, Narender Tadigoppula
P735 Circular disulfide-‐rich peptide scaffolds as anti-‐citrullinated peptide antibody inhibitors
Sunithi Gunasekera, Catia Fernandes-‐Cerqueira, Camilla Eriksson, Per-‐Johan Jakobsson, Ulf Göransson
P736 Engineering of KR-‐12: A minimalized domain derived from human host defense peptide LL-‐37
into a potent antimicrobial drug lead
Taj Muhammad, Sunithi Gunasekera, Adam A. Strömstedt, Ulf Göransson
P737 In silico identification and in vitro activity of new pancreatic lipase inhibitors
Tina Buchholz , Anne Frank, Gerhard Wolber, Matthias F. Melzig
P738 Anti-‐HIV natural products (28): preparation of conjugate for 3-‐O-‐acyl betulin derivative and
AZT as anti-‐HIV agents
Shizuka Wada, Naonobu Tanaka, Chin-‐Ho Chen, Susan L. Morris-‐Natschke, Kuo-‐Hsiung Lee, Yoshiki
Kashiwada
P739 Exploring natural coumarin derivatives as agents against Mycobacterial biofilm, highlighting
inhibition and eradication
Carel Basson Oosthuizen, Navneet Kishore, Namrita Lall
Abstract book
• Plenary lectures
• Keynote lectures
• Short lectures
• Young Researchers Workshop
• Pre-conference symposium
• Poster session Monday
• Poster session Tuesday
• Poster session Wednesday
P366
Chemical
characterization
and
in
vitro
antibacterial
activity
of
Kalanchoe
brasiliensis
Cambess
(Crassulaceae)
Oscar
A.
S.
Mayorga1,
Jônatas
R.
Florencio1,
Jordana
D.
G.
de
Santana1,
Carolina
Feres-‐Netto2,
Dionnata
M.
Pedrosa2,
Ygor
F.
G.
Costa2,
Nícolas
C.
C.
Pinto3,
Elita
Scio3,
Orlando
V.
Sousa4,
Ma-‐
ria
Silvana
Alves4
1Graduate
Program
in
Pharmaceutical
Sciences,
Universidade
Federal
de
Juiz
de
Fora,
Rua
José
Lourenço
Kelmer,
s/n,
Campus
Universitário,
São
Pedro,
CEP
36036-‐900,
Juiz
de
Fora,
Minas
Gerais,
Brazil,
2Pharmacy
Course,
Faculty
of
Pharmacy,
Universidade
Federal
de
Juiz
de
Fora,
Rua
José
Lourenço
Kelmer,
s/n,
Campus
Universitário,
São
Pedro,
CEP
36036-‐900,
Juiz
de
Fora,
Brazil,
3Laboratory
of
Bioactive
Nat-‐
ural
Products,
Department
of
Biochemistry,
Federal
University
of
Juiz
de
Fora,
36036-‐900,
Juiz
de
Fora,
MG,
Brazil,
4Department
of
Pharmaceutical
Sciences,
Faculty
of
Pharmacy,
Universidade
Federal
de
Juiz
de
Fora,
Rua
José
Lourenço
Kelmer,
s/n,
Campus
Universitário,
São
Pedro,
CEP
36036-‐900,
Juiz
de
Fora,
Minas
Gerais,
Brazil.
Kalanchoe
brasiliensis
Cambess
(Crassulaceae),
popularly
known
as
"Saião",
is
a
Brazilian
me-‐
dicinal
plant
distributed
from
Bahia
to
São
Paulo
and
traditionally
used
to
treat
injuries,
ab-‐
scesses,
enlarged
ganglia
and
inflammatory
processes
[1,2].
The
current
study
aimed
to
char-‐
acterize
the
chemical
composition
of
the
ethanolic
extract
50%
(v/v)
obtained
from
K.
brasili-‐
ensis
leaves
collected
before
blooming
using
HPLC-‐DAD
[3]
and
to
investigate
the
antibacterial
activity
of
this
extract
at
different
concentrations
(30%,
50%
and
70%).
The
chromatography
profile
and
the
UV
spectrum
analysis
suggested
the
significant
presence
of
flavonoids.
The
antibacterial
activity
was
established
according
to
the
Clinical
Laboratory
Standards
Institute
(CLSI)
guidelines
by
the
Minimal
Inhibitory
Concentration
(MIC)
[4]
using
the
microdilution
method
followed
by
the
Minimal
Bactericidal
Concentration
(MBC),
which
allows
to
classify
the
antibacterial
effect
as
bacteriostatic
or
bactericidal
[5].
Staphylococcus
aureus
subsp.
au-‐
reus
(ATCC®
29213™),
Staphylococcus
aureus
subsp.
aureus
(ATCC®
6538™),
Pseudomonas
aeruginosa
(ATCC®
9027™),
Pseudomonas
aeruginosa
(ATCC®
27853™),
Escherichia
coli
(ATCC®25922™),
Escherichia
coli
(ATCC®10536™),
Salmonella
enterica
subsp.
enterica
serovar
Typhimurium
(ATCC®
13311™)
and
Salmonella
enterica
subsp.
enterica
serovar
Chol-‐
eraesuis
(ATCC®
10708™)
were
used
as
reference
strains.
The
ethanolic
extract
at
30%
and
50%
demonstrated
bacteriostatic
effect
against
S.
Typhimurium
ATCC®
13311™
and
S.
Chol-‐
eraesuis
ATCC®
10708™,
with
MIC
values
of
5000
µg/mL.
Additionally,
the
ethanolic
extract
at
70%
revealed
bacteriostatic
effect
against
S.
aureus
ATCC®
6538™
and
ATCC®25922™
and
against
S.
Thyphimurium
ATCC®
13311™
and
S.
Choleraesuis
ATCC®
10708™,
with
MIC
values
of
5000
µg/mL,
showing
a
broad
spectrum
of
action.
These
results
suggest
that
K.
brasiliensis
can
be
an
interesting
source
of
bioactive
molecules
with
antibacterial
potential.
Acknowledgements:
This
investigation
was
supported
by
UFJF,
FAPEMIG,
CAPES,
OEA
and
GCUB
Keywords: Kalanchoe brasiliensis, Flavonoids, HPLC-‐DAD, Anti-‐Bacterial Agents, Medicinal
Plants
References:
[1] Costa
SS,
Jossang
A,
Bodo
B,
Souza
ML,
Moraes
VL.
Patuletin
acetylrhamnosides
from
Kal-‐
anchoe
brasiliensis
as
inhibitors
of
human
lymphocyte
proliferative
activity.
J
Nat
Prod
1994;
57:
1503-‐1510
[2] Mourão
RH,
Santos
FO,
Franzotti
EM,
Moreno
MP,
Antoniolli
AR.
Antiinflammatory
activity
and
acute
toxicity
(LD50)
of
the
juice
of
Kalanchoe
brasiliensis
(Camb.)
leaves
picked
be-‐
fore
and
during
blooming.
Phytother
Res
1999;
13:
352-‐354.
[3] Costa
ACO,
Fernandes
JM,
Themístocles
NNS,
Mendonça
JN,
Tomaz
JC,
Lopes
NP,
Alberto
L,
Soares
L,
Zucolotto
SM.
Quantification
of
chemical
marker
of
Kalanchoe
brasiliensis
(Cras-‐
sulaceae)
leaves
by
HPLC-‐DAD.
J
Liq
Chromatogr
Rel
Technol
2015,
38:
795-‐800
[4] Clinical
and
Laboratory
Standards
Institute.
Methods
for
Dilution
Antimicrobial
Suscepti-‐
bility
Tests
for
Bacteria
That
Grow
Aerobically;
Approved
Standard
-‐
Ninth
Edition
(M07-‐
A9).
Wayne:
CLSI,
2012
[5] Andrews
JM.
Determination
of
minimum
inhibitory
concentrations.
J
Antimicrob
Chemother
2001;
48:
5-‐16
P367
In
vitro
antibacterial
activity
of
Vernonia
polyanthes
Less.
leaf
rinse
extract
(Asteraceae):
prospecting
new
therapeutic
options
against
Staphylococcus
aureus
infections!
Jordana
D.
G.
de
Santana1,
Jonatas
R.
Florêncio1,
Luísa
M.
S.
de
Almeida1,
Laura
S.
Fernandes2,
Ademar
A.
da
Silva
Filho3,
Marcos
José
Salvador4,
Orlando
V.
Sousa3,
Maria
Silvana
Alves3
1Graduate
Program
in
Pharmaceutical
Sciences,
Faculty
of
Pharmacy,
Universidade
Federal
de
Juiz
de
Fora,
Rua
José
Lourenço
Kelmer,
s/n,
Campus
Universitário,
São
Pedro,
CEP
36036-‐900,
Juiz
de
Fora,
Bra-‐
zil,
2Pharmacy
Course,
Faculty
of
Pharmacy,
Universidade
Federal
de
Juiz
de
Fora,
Rua
José
Lourenço
Kelmer,
s/n,
Campus
Universitário,
São
Pedro,
CEP
36036-‐900,
Juiz
de
Fora,
Brazil,
3Department
of
Pharmaceutical
Sciences,
Faculty
of
Pharmacy,
Universidade
Federal
de
Juiz
de
Fora,
Rua
José
Lourenço
Kelmer,
s/n,
Campus
Universitário,
São
Pedro,
CEP
36036-‐900,
Juiz
de
Fora,
Brazil,
4Department
of
Vege-‐
table
Biology,
Biology
Institute,
Universidade
Estadual
de
Campinas,
Rua
Monteiro
Lobato,
255,
CEP
13083-‐862,
Campinas,
Brazil
Vernonia
polyanthes
Less.
(Asteraceae),
popularly
known
as
"assa-‐peixe",
is
a
native
vegetal
species
of
South
America,
especially
in
Brazil
[1].
This
medicinal
plant
is
traditionally
used
in
cases
of
cold,
flu,
cough,
fever,
bronchitis
and
pneumonia
[2].
Anatomically,
V.
polyanthes
pre-‐
sents
glandular
trichomes,
which
are
modified
epidermal
hairs
that
contain
specialized
cells
involved
in
the
biosynthesis,
storage
and
secretion
of
a
number
of
natural
molecules,
includ-‐
ing
secondary
metabolites
[1,3].
Flavonoids
and
sesquiterpene
lactones
are
considered
chem-‐
ical
markers
of
the
Vernonia
genus
[4].
The
current
study
investigated
the
in
vitro
antibacteri-‐
al
activity
of
V.
polyanthes
leaf
rinse
extract
(Vp-‐LRE)
in
order
to
corroborate
with
the
tradi-‐
tional
use
of
this
plant
species
and
its
potential
as
a
source
of
new
antibacterial
agents.
In
vitro
antibacterial
activity
was
established
by
the
Minimal
Inhibitory
Concentration
(MIC)
using
the
microdilution
method
according
to
the
Clinical
Laboratory
Standards
Institute
(CLSI)
guide-‐
lines
[5]
followed
by
the
Minimal
Bactericidal
Concentration
(MBC),
which
allows
classifica-‐
tion
of
the
antibacterial
effect
as
bacteriostatic
or
bactericidal
[6].
Staphylococcus
aureus
ATCC®
6538™
and
ATCC®
29213™,
Escherichia
coli
ATCC®
10536™
and
ATCC®
25922™,
Sal-‐
monella
enterica
subsp.
enterica
serovar
Choleraesuis
ATCC®
10708™,
Salmonella
enterica
subsp.
enterica
serovar
Thyphimurium
ATCC®
13311™
and
Pseudomonas
aeruginosa
ATCC®
9027™
and
ATCC®
27853™
were
used
as
reference
strains.
Vp-‐LRE
revealed
antibacterial
ac-‐
tivity
against
5
of
8
tested
strains,
being
more
active
against
S.
aureus
ATCC®
6538™
and
ATCC®
29213™
with
MIC
values
of
625
µg/mL
for
both,
demonstrating
bacteriostatic
effect
and
MBC
of
2500
µg/mL
and
1250
µg/mL,
respectively.
These
results
suggest
the
potential
of
V.
polyanthes
as
a
promising
source
of
bioactive
molecules
with
antibacterial
action,
prospect-‐
ing
new
therapeutic
approaches
against
S.
aureus
infections.
Acknowledgements:
This
investigation
was
financially
supported
by
UFJF,
FAPEMIG,
CNPq
and
CAPES
Keywords: Vernonia polyanthes, Asteraceae, Glandular Trichomes, Antibacterial Agents,
Staphylococcus aureus
References:
[1] Alves
VFG,
Neves
LJ.
Anatomia
foliar
de
Vernonia
polyanthes
Less.
(Asteraceae).
Rev
Univ
Rural,
Sér
Ciên
da
Vida
2003;
22:
1-‐8
[2] Rodrigues
VEG,
Carvalho,
DA.
Levantamento
etnobotânico
de
plantas
medicinais
no
domí-‐
nio
do
cerrado
na
região
do
Alto
Rio
Grande
-‐
Minas
Gerais.
Ciênc
Agrotec
2001;
25:
102-‐
123
[3] Lange
BM,
Turner,
GW.
Terpenoid
biosynthesis
in
trichomes
-‐
current
status
and
future
opportunities.
Plant
Biotech
J
2013;
11:
2-‐22
[4] Costa
FJ,
Bandeira
PN,
Albuquerque
MRJR,
Pessoa
ODL,
Silveira
ER,
Braz-‐filho
R.
Constitu-‐
intes
químicos
de
Vernonia
chalybaea
Mart.
Quim
Nova
2008;
31:
1691-‐1695.
[5] Andrews
JM.
Determination
of
minimum
inhibitory
concentrations.
J
Antimicrob
Chemother
2001;
48:
5-‐16
[6] Clinical
and
Laboratory
Standards
Institute.
Methods
for
Dilution
Antimicrobial
Suscepti-‐
bility
Tests
for
Bacteria
That
Grow
Aerobically;
Approved
Standard
-‐
Ninth
Edition
(M07-‐
A9).
Wayne:
CLSI,
2012
P368
Steroidal
saponins
from
Chlorophytum
deistelianum
Turibio
Tabopda1,2,
Anne-‐Claire
Mitaine-‐Offer1,
Thomas
Paululat3
,
Stéphanie
Delemasure4,
Patrick
Dutartre4,
Bonaventure
Tchaleu
Ngadjui2,
Marie-‐Aleth
Lacaille-‐Dubois1
1Laboratoire
de
Pharmacognosie,
EA
4267
FDE,
Université
de
Bourgogne
Franche-‐Comté,
7,
Bd.
Jeanne
d’Arc,
BP
87900,
21079
Dijon
Cedex,
France,
2Département
de
Chimie
Organique,
Université
de
Yaoundé
1,
BP
812
Yaoundé,
Cameroun,
3Universität
Siegen,
OC-‐II,
Naturwissenschaftlich-‐Technische
Fakultät,
Adolf-‐Reichwein-‐Strasse,
D-‐57076
Siegen,
Germany,
4Cohiro,
UFR
des
Sciences
de
Santé,
7,
Bd.
Jeanne
d'Arc,
BP
87900,
21079
Dijon
Cedex,
France.
Plants
of
the
genus
Chlorophytum
are
known
as
a
rich
source
of
steroidal
saponins[1-‐3]
which
have
attracted
the
attention
for
their
structural
diversity
and
significant
bioactivities
such
as
cytotoxic,
immunomodulating,
antifungal,
insecticidal
activities,
just
to
mention
a
few
[4].
In
our
continuing
search
for
bioactive
saponins
from
the
genus
Chlorophytum[1-‐3]
we
investi-‐
gated
Chlorophytum
deistelianum
Engl.
&
Krause
(=
C.
sparsiflorum
Backer
var.
sparsiflorum)
from
a
phytopharmacological
point
of
view.
C.
deistelianum
is
a
fleshy
herb
to
50
cm
high,
with
white
or
greenish
flowers.
No
study
on
the
secondary
metabolites
of
this
plant
has
been
re-‐
ported
so
far.
Therefore,
we
report
herein
the
isolation
of
four
previously
undescribed
steroi-‐
dal
saponins,
called
chlorodeistelianosides
A
–
D
(1-‐4)
together
with
five
known
steroidal
saponins
from
C.
desteilianum.
Their
structures
were
determined
by
spectroscopic
methods
including
1D
and
2D
NMR
experiments,
ESI
and
HRESIMS.
Compounds
1
and
2
are
glycosides
of
hecogenin
and
its
24β-‐OH
derivative,
respectively,
whereas
3
is
a
glycoside
of
(25R)-‐5α-‐
22α-‐methoxyfurostane-‐2α,3β,26
triol
and
4
a
glycoside
of
(25R)-‐5α-‐furost-‐20(22)-‐en-‐12-‐
one-‐3β,26
diol.
Compounds
2-‐4
share
the
same
sugar
sequence
at
C-‐3
characterized
as
β-‐D-‐
glucopyranosyl-‐(1→2)-‐[β-‐D-‐xylopyranosyl-‐(1→3)]-‐β-‐D-‐glucopyranosyl-‐(1→4)-‐β-‐D-‐
galactopyranosyl
whereas
the
sugar
sequence
of
1
was
elucidated
as
β-‐D-‐glucopyranosyl-‐
(1→3)-‐[α-‐L-‐rhamnopyranosyl-‐(1→4)]-‐β-‐D-‐xylopyranosyl-‐(1→3)-‐[β-‐D-‐glucopyranosyl-‐
(1→2)]-‐β-‐D-‐glucopyranosyl-‐(1→4)-‐β-‐D-‐galactopyranosyl.
Furthermore,
seven
compounds
were
examined
for
cytotoxicity
against
one
human
colorectal
adenocarcinoma
cell
line
(SW480)
and
one
rat
cardiomyoblast
cell
line
(H9c2).
Among
them,
three
known
spirostane-‐
type
glycosides
exhibited
cytotoxicity
on
both
cell
lines
with
IC50
ranging
from
8
to
10
µM.
Keywords: Chlorophytum deistelianum, Asparagaceae, steroid saponins, NMR, cytotoxicity
References:
[1] Acharya
D,
Mitaine-‐Offer
A-‐C,
Kaushik
N,
Miyamoto
T,
Paululat
T,
Lacaille-‐Dubois
M-‐A.
Fu-‐
rostane-‐type
steroidal
saponins
from
the
roots
of
Chlorophytum
borivilianum.
Helv
Chim
Acta
2008;
91:
2262–2269
[2] Acharya
D,
Mitaine-‐Offer
A-‐C,
Kaushik
N,
Miyamoto
T,
Paululat
T,
Mirjolet
JF,
Duchamp
O,
Lacaille-‐Dubois
M-‐A.
Cytotoxic
spirostane-‐type
saponins
from
the
roots
of
Chlorophytum
borivilianum.
J
Nat
Prod
2009;
72:
177–181
[3] Acharya
D,
Mitaine-‐Offer
A-‐C,
Kaushik
N,
Miyamoto
T,
Paululat
T,
Mirjolet
JF,
Duchamp
O,
Lacaille-‐Dubois
M-‐A.
Steroidal
saponins
from
Chlorophytum
orchidastrum.
J
Nat
Prod
2010;
73:
7–11.
[4] Lacaille-‐Dubois
M-‐A.
Bioactive
saponins
with
cancer
related
and
immunomodulatory
ac-‐
tivity:
recent
developments.
In:
Atta-‐Ur-‐Rahman,
editor.
Studies
in
Natural
Products
Chemistry
series.
Amsterdam:
Elsevier,
2005;
32:
209–246
P369
Triterpene
glycosides
from
plants
for
antibody
recognition
Elisa
Peroni1,3,
Feliciana
Real
Fernández1,2,
Caterina
Gheri1,2,
Francesca
Nuti1,2,
Anne-‐Claire
Mitaine-‐Offer4,
Francesco
Lolli1,5,
Marie-‐Aleth
Lacaille-‐Dubois4,
Anna-‐Maria
Papini1,2,3
1
Laboratory
of
Peptide
and
Protein
Chemistry
and
Biology,
University
of
Florence,
50019
Sesto
Fiorenti-‐
no,
Italy,
2
Department
of
Chemistry
“Ugo
Schiff”
and
NEUROFARBA
Department,
section
of
Pharmaceuti-‐
cal
Sciences
and
Nutraceutics,
University
of
Florence,
50019
Sesto
Fiorentino,
Italy,
3
Laboratory
of
Chem-‐
ical
Biology
EA
4505
&
PeptLab@UCP,
Université
de
Cergy-‐Pontoise,
95031
Cergy-‐Pontoise
cedex,
France,
4
Laboratoire
de
Pharmacognosie,
EA
4267
FDE,
Université
de
Bourgogne-‐Franche-‐Comté,
21079
Dijon
cedex,
France,
5
Department
of
Biomedical,
Experimental
and
Clinical
Sciences,
University
of
Florence,
50134
Firenze,
Italy
Multiple
sclerosis
is
an
autoimmune
disease
that
affects
the
central
nervous
system.
The
key
role
of
glycosylation
in
disease
pathogenesis
has
been
studied
previously
and
the
synthetic
N-‐
glucosylated
peptide
CSF114
(Glc)
proved
its
efficiency
in
autoantibody
recognition
in
sera
of
multiple
sclerosis
patients
[1].
Therefore,
it
was
interesting
to
test
natural
compounds
such
as
triterpene
glycosides
from
plants,
possessing
a
wide
variety
of
glycosyl
moieties
for
autoanti-‐
body
recognition.
Five
molecules,
isolated
from
different
plants
were
tested
in
sera
from
mul-‐
tiple
sclerosis
patients
to
better
understand
the
role
of
glycosylation:
two
presenegenin
glyco-‐
sides
from
the
Polygalaceae
family
[2],
two
sulfated
quinovic
acid
glycosides
from
the
Zygo-‐
phyllaceae
family
[3],
and
one
cycloartane
glycoside
from
the
Asteraceae
family
[4].
The
in-‐
teraction
between
these
glycosides
and
autoantibodies
was
evaluated
by
ELISA
measuring
IgG
and
IgM
levels
in
multiple
sclerosis
patients
and
healthy
blood
donors,
and
results
were
com-‐
pared
with
those
for
CSF114(Glc).
The
ursolic
acid
was
chosen
to
evaluate
the
activity
of
the
aglycon
only
and
thus
the
role
of
the
oligosaccharidic
part
of
the
molecule.
The
five
natural
triterpene
glycosides
showed
good
capacity
to
recognize
IgMs
(sensitivity
up
to
38%)
with
high
specificity
(88.9%)
for
multiple
sclerosis
patients
[5].
According
to
the
biological
results,
the
best
structures
to
be
selected
have
been
deduced:
monodesmosidic
triterpene
glycosides
with
only
one
glucopyranosyl
moiety
in
position
C-‐3
or
bidesmosidic
with
one
sugar,
sulfated
or
not,
at
the
C-‐3
position
and
a
substitution
with
only
one
glucopyranosyl
moiety
at
the
C-‐24
or
C-‐28
position.
The
ursolic
acid
presenting
the
scaffold
of
the
five
glycosides
but
lacking
the
glycosyl
moieties
was
not
able
to
identify
specific
MS
antibodies.
Keywords:
Multiple
sclerosis,
triterpene
glycosides,
autoimmune
disease,
biomarkers,
auto-‐
antibody
recognition
References:
[1] Carotenuto
A,
Alcaro
MC,
Saviello
MR,
Peroni
E,
Nuti
F,
Papini
AM,
Novellino
E,
Rovero
P.
Designed
glycopeptides
with
different
β-‐turn
types
as
synthetic
probes
for
the
detection
of
autoantibodies
as
biomarkers
of
multiple
sclerosis.
J
Med
Chem
2008;
51:
5304–5309
[2] Lacaille-‐Dubois
M-‐A,
Mitaine-‐Offer
A-‐C.
Triterpene
saponins
from
Polygalaceae.
Phyto-‐
chemistry
Rev
2005;
4:
139–149
[3] Smati
D,
Mitaine-‐Offer
A-‐C,
Miyamoto
T,
Hammiche
V,
Lacaille-‐Dubois
M-‐A.
Ursane-‐type
triterpene
saponins
from
Zygophyllum
geslini.
Helv
Chim
Acta
2007;
90:
712–719
[4] Mitaine-‐Offer
A-‐C,
Miyamoto
T,
Semmar
N,
Jay
M,
Lacaille-‐Dubois
M-‐A.
A
new
oleanane
gly-‐
coside
from
the
roots
of
Astragalus
caprinus.
Magn
Reson
Chem
2006;
44:
713–716
[5] Peroni
E,
Real
Fernández
F,
Gheri
C,
Nuti
F,
Mitaine-‐Offer
A-‐C,
Lolli
F,
Lacaille-‐Dubois
M-‐A,
Papini
AM.
Natural
triterpene
glycosides
for
antibody
recognition.
Planta
Med
Lett
2016;
3:
e1–e6
P370
Chemical
characterisation
and
cytotoxicity
evaluation
of
Convolvulus
pluricaulis
Sieb.
ex
Spreng.
(Convolvulaceae)
extracts
towards
sensitive
and
multidrug-‐resistant
cancer
cells
Massimo Tacchini1, Guglielmo Paganetto1, Thomas Efferth2, Gianni Sacchetti1, Alessandra
Guerrini1
1 Department of Life Sciences and Biotechnology, University of Ferrara, Corso Ercole I° d'Este 32, 44121
Ferrara,
Italy,
2
Department
of
Pharmaceutical
Biology,
Institute
of
Pharmacy
and
Biochemistry
–
Johannes
Gutenberg
University,
Mainz,
Germany
The
modern
research
on
cancer
prevention
and
therapy
is
looking
at
the
plant
kingdom
as
possible
source
of
molecules
or
new
molecular
platforms.
Given
the
large
number,
variety,
and
the
wide
complexity
of
molecules
synthesised
by
plant
cells,
researches
investigate
ethnopharmacology
with
the
aim
of
enhancing
the
probability
to
discover
new
effective
drugs
[1].
In
relation
to
these
premises,
two
traditional
preparations
of
Convolvulus
pluricaulis
Sieb.
ex
Spreng.
(Convolvulaceae)
whole
plant
was
investigated:
decoction
(DEC),
derived
from
the
Ayurvedic
medicinal
system;
and
hydro-‐alcoholic
extract
(HE)
closer
to
the
Western
phytotherapic
tradition.
The
chemical
fingerprinting
showed
two
preparations
characterized
by
the
presence
of
phenolic
compounds
(es.
caffeic
acid,
p-‐coumaric
acid,
isoferulic
acid,
ferulic
acid
and
vanillin),
identified
and
quantified
by
gas
chromatography
following
a
full
validated
method
[2].
DEC,
moreover,
showed
the
presence
of
stigmasterol
and
lupeol.
Taking
into
account
the
biological
activity
of
C.
pluricaulis
reported
in
literature,
and
with
the
aim
of
extending
its
bioactivity
panorama,
the
cytotoxic
effect
of
the
preparations,
fractions
and
molecules,
against
two
leukemic
cell
lines,
one
drug-‐sensitive
(CCRF-‐CEM)
and
one
multi-‐
drug-‐resistant
(CEM/ADR5000)
was
investigated.
The
best
result
against
the
CCRF
cells
was
exhibited
by
the
CHCl3
DEC
extract
(IC50=
16,18±0,79
µg/ml),
it
could
be
explained
by
the
presence
of
lupeol
(IC50=
9,62±0,21
μM),
molecule
with
a
well-‐known
anticancer
activity.
ADR5000,
instead,
showed
cross-‐resistance
towards
the
every
tested
phytocomplexes
and
molecules,
even
if
they
exhibited
IC50
values
lower
than
the
doxorubicin
one.
The
next
step
of
the
research
will
be
the
ADME
evaluation
of
identified
molecules
using
in
silico
model
(GastroPlus™),
and
the
assessment
of
the
phytocomplexes
activity
agains
cancer
stem
cells.
Keywords: Convolvulus pluricaulis, chemical characterisation, cytotoxicity, multidrug
resistance, leukemia
References:
[1] Guerrini
A,
Sacchetti
G.
From
Ethnobotany
towards
Modern
Botanicals
as
Paradigm
of
the
Medicinal
Plants
Research:
The
Case
of
Ayurveda.
Med
Aromat
Plants
2012;
1
[2] Caligiani
A,
Malavasi
G,
Palla
G,
Angela
Marseglia
A,
Massimiliano
Tognolini
M,
Bruni
R.
A
simple
GC–MS
method
for
the
screening
of
betulinic,
corosolic,
maslinic,
oleanolic
and
ursolic
acid
contents
in
commercial
botanicals
used
as
food
supplement
ingredients.
Food
Chem
2013;
136:
735-‐741
P371
Survey
of
folk
use
and
gathering
of
plants
in
Karst
and
Gorjanci,
Slovenia
Mateja Lumpert1, Samo Kreft1
1 Chair of Pharmaceutical Biology, University of Ljubljana, Faculty of Pharmacy, Aškerčeva cesta 7, SI-‐
1000 Ljubljana, Slovenia
Folk
use
and
gathering
of
plants
in
nature
was
investigated
in
Karst
and
Gorjanci,
Slovenia.
From
October
2013
to
September
2014,
25
informants
in
each
region
were
interviewed.
Their
average
age
was
61
years
in
Karst
and
68
in
Gorjanci.
The
main
question
was:
"Which
plants
do
you
gather
or
did
you
gather
in
nature
and
use?"
Plants
with
medicinal,
veterinary,
cosmet-‐
ic
or
nutritive
uses
were
considered.
A
total
of
78
taxa
were
reported
in
Karst
and
82
in
Gor-‐
janci;
65
taxa
were
mutual
in
both
regions.
Data
suggest
that
there
are
minor
the
differences
in
gathering
of
plants
between
Karst
and
Gorjanci
and
they
are
mostly
due
to
the
natural
habi-‐
tat
of
some
plants;
on
the
other
hand,
regional
differences
in
medicinal
use
of
some
commonly
gathered
plants
were
observed,
e.g.
19
informants
in
Gorjanci
(G:
19)
and
16
in
Karst
(K:
16)
gathered
Chamomilla
recutita.
While
most
Karst
informants
used
it
as
calmative
(G:
1;
K:
7),
most
of
Gorjanci
informants
used
it
as
carminative
(G:
13;
K:
5),
respiratory
infections
(G:
6;
K:
1),
sore
eyes
(G:
9;
K:
2)
and
dysmenorrhea
(G:
4;
K:
1).
Sambucus
nigra
was
the
most
fre-‐
quently
gathered
plant
(G:
24;
K:
24).
While
most
of
Gorjanci
informants
used
it
for
treatment
of
respiratory
infections
(G:
14;
K:
5),
most
of
Karst
informants
used
it
merely
for
nutritive
purposes
(G:
10;
K:
18).
Altogether
16
different
medicinal
preparations
for
oral
application
were
reported
and
15
for
topical
application
(skin,
ears,
eyes
and
lungs).
Unusual
medicinal
preparations
were
observed:
browned
flour
soup
(prežganka
in
Slovene)
and
egg
omelet
(frtalja
in
Slovene).
They
were
prepared
with
bitter
herbal
plants
(Achillea
millefolium,
Ruta
graveolens,
Tanacetum
vulgare
and
C.
recutita)
and
used
for
treatment
of
gastrointestinal
problems.
As
a
source
of
knowledge
about
plants,
21
informants
in
Gorjanci
and
20
in
Karst
obtained
oral
information
from
at
least
one
person.
Books
were
reported
by
10
informants
in
Gorjanci
and
20
in
Karst.
Acknowledgements:
Slovenian
Research
Agency
(ARRS)
Keywords: Karst, Gorjanci, Chamomilla recutita, Sambucus nigra, Ethnobotany.
P372
Anti-‐inflammatory
effects
and
toxicity
of
polysaccharide
fraction
from
Khaya
grandifoliola
stem
bark
Mediesse
Kengne
Francine1,
Hasitha
Anantharaju3,
Gangadhar
Matharasala3,
Mbacham
F.
Wil-‐
fried2,
Boudjeko
Thaddée1,2,
Yogeeswari
Perumal3
1Laboratory
of
Phytoprotection
and
Valorization
of
Plant
Resources,
Biotechnology
Centre,
University
of
Yaounde
I,
BP
3851,
Messa-‐Yaounde
Cameroon;
2Department
of
Biochemistry,
University
of
Yaounde
I,
BP
812,
Yaounde,
Cameroon;
3Drug
Discovery
Research
Lab,
Department
of
Pharmacy,
Birla
Institute
of
Technology
&
Science–Pilani,
Hyderabad
campus,
Jawahar
Nagar,
Hyderabad–500078,
Andhra
Pradesh,
India
Traditional
medicine
occupies
an
important
position
in
the
prevention
and
treatment
of
dis-‐
eases
due
their
availability
and
low
cost.
Despite
the
possible
presence
of
polysaccharides
in
decoctions,
infusions,
and
macerations
few
studies
have
look
the
role
of
these
metabolites
in
the
efficacy
of
these
medicinal
plants.
The
aim
of
the
study
was
to
investigate
the
anti-‐
inflammatory
and
toxic
effects
of
polysaccharide
fraction
(lG)
of
stem
bark
of
Khaya
grandifo-‐
liola
C.
DC.
(Meliaceae),
which
is
mostly
used
by
the
population
and
traditional
healers
in
Afri-‐
ca
to
treat
many
diseases
like
malaria
and
fever.
Extraction
was
performed
by
boiling
in
hot
water
and
precipitation
in
alcohol
(1:2
v/v).
The
fraction
was
de-‐proteinsed
with
Sevag
rea-‐
gent
and
lyophilized.
The
cytotoxicity
was
evaluated
on
RAW
264.7
macrophages
and
Brain
glioblastoma
U87MG
cells
line
by
MTT
assay.
Our
results
showed
that
lG
is
non-‐toxic
on
the
two
cell
lines
tested
(IC50
>
1mg/mL).
More
over,
the
ability
of
lG
to
inhibit
the
LPS-‐induced
overproduction
of
NO
by
macrophages
and
ROS
by
U87MG
were
tested
using
Griess
reaction
and
DCFA-‐DCF
assay,
respectively
[1,
2].
In
addition,
the
ability
of
this
fraction
to
down
regu-‐
late
the
LPS-‐induced
over-‐expression
of
pro-‐inflammatory
mediators
NF-‐kB,
IL6,
IL1β,
and
TNFα
genes
was
evaluated
in
vitro
on
U87
cells
and
in
vivo
on
brain
of
treated
Balb
c
mice
by
quantitative
Real
time
PCR.
This
last
experiment
was
perfomed
to
confirm
that
lG
at
the
test
concentration
do
not
have
neurotoxic
(on
locomotor
activities
and
muscle
coordination)
ef-‐
fect
on
mice.
The
results
showed
that
lG
significantly
(p
<
0.001)
inhibits
the
over-‐expression
of
these
genes
stimulated
by
LPS
in
vitro
and
in
vivo.
These
findings
stand
as
baseline
data
for
future
studies,
which
will
help
in
the
field
of
research
on
adjuvant
based
polysaccharide
used
in
the
treatment
of
inflammation-‐related
diseases
in
humans.
However,
further
investigation
of
physico-‐chemical
properties
and
structure
elucidation
of
the
polysaccharide
is
required
to
understand
the
mechanism
involved.
Acknowledgements:
We
gratefully
acknowledge
the
NAM
S&T
Centre
for
providing
the
RTF-‐DCS
14-‐15
fellowship
at
BITS,
pilani
(India)
to
one
of
the
authors
Mediesse
Kengne
Francine.
Financial
support
by
TWAS
Research
Grant
Program
in
Basic
Sciences
is
also
gratefully
acknowledged.
Keywords: Khaya grandifoliola, polysaccharide fraction, anti-‐inflammatory, toxicity
References:
[1] Wu
D,
Yotnda
P.
Production
and
detection
of
reactive
oxygen
species
(ROS)
in
cancers.
J
Visual
Exp
2011;
57:
2–5
[2] Cheenpracha
S,
Rostama
B,
Pezzuto
JM,
Chang
LC.
Inhibition
of
nitric
oxide
(NO)
production
in
RAW
264.7
cells
by
the
norsesterterpene
peroxide,
epimuqubilin
A.
Mar
Drugs
2010;
8:
429-‐437
P373
Chemical
constituents
from
Agrimonia
pilosa
with
their
protein
tyrosine
phosphatase
and
acetylcholinesterase
inhibitory
activi-‐
ties
Duc
H.
Nguyen1,2,
Duc
D.
Le1,
U
M.
Seo1,
Thi
T.
Nguyen1,
Jae Soo Choi3,
Mi
H.
Woo
1
1College
of
Pharmacy,
Catholic
University
of
Daegu,
Gyeongsan
38430,
Republic
of
Korea;
2Phutho
College
of
Pharmacy,
Viettri
City,
Phutho
Province
290000,
Vietnam;
3Department
of
Food
Science
&
Nutrition,
Pukyong
National
University,
Busan
608-‐737,
Republic
of
Korea
Agrimonia
pilosa
Ledeb.,
a
perennial
herb,
belongs
to
the
Rosaceae
family.
The
whole
plant
A.
pilosa
has
been
used
traditionally
as
hemostatic,
antimalarial,
and
antidysenteric
in
Chinese
herbal
medicine
for
a
long
time
[1,2].
Two
new
flavanonol
glucoside
isomers,
(2S,3R)
dihy-‐
drokaempferol
3-‐O-‐β-‐ᴅ-‐glucoside
(1)
and
(2R,3S)
dihydrokaempferol
3-‐O-‐β-‐ᴅ-‐glucoside
(2),
were
isolated
from
the
aerial
parts
of
A.
pilosa,
along
with
12
known
compounds
(3‒14).
Their
structures
were
determined
on
the
basis
of
spectroscopic
analyses.
In
addition,
all
the
isolates
were
evaluated
for
PTP1B
inhibitory
activity.
Among
them,
compounds
13
and
14
displayed
potential
inhibitory
activity
against
PTP1B
with
IC50
values
of
7.14
±
1.75
and
7.73
±
0.24
μM,
respectively.
Furthermore,
all
the
isolated
compounds
were
found
to
inhibit
AChE
with
IC50
values
ranging
from
62.96
±
0.35
to
118.32
±
0.09
μM.
Keywords:
Agrimonia
pilosa,
Flavanonol
glucosides,
Protein
tyrosine
phosphatase
1B,
acetyl-‐
cholinesterase
References:
[1] Kim
JJ,
Jiang
J,
Shim
DW,
Kwon
SC,
Kim
TJ,
Ye
SK,
Kim
MK,
Shin
YK,
Koppula
S,
Kang
TB,
Choi
DK,
Lee
KH.
Anti-‐inflammatory
and
anti-‐allergic
effects
of
Agrimonia
pilosa
Ledeb
extract
on
murine
cell
lines
and
VOA-‐induced
airway
inflammation.
J
Ethnopharmacol
2012;
140:
213-‐221
[2] Taira
J,
Nanbu
H,
Ueda
K.
Nitric
oxide-‐scavenging
compounds
in
Agrimonia
pilosa
Ledeb
on
LPS-‐induced
RAW264.7
macrophages.
Food
Chem
2009;
115:
1221-‐1227
P374
Simultaneous
quantitation
of
five
bioactive
components
of
Men-‐
thae
Herba
by
HPLC/PDA
Mi
Hee
Woo1,
Bing
Tian
Zhao1,
Min
Je
Choi1,
Young
Ho
Kim2,
Jong
Seong
Kang2.
1
College
of
Pharmacy,
Catholic
University
of
Daegu,
Gyeongsan,
38430,
Republic
of
Korea,
2
College
of
Pharmacy, Chungnam National University, Daejeon, 34134, Republic of Korea
Several
different
methods
have
been
developed
to
qualify
and
quantitate
the
components in
Menthae
Herba
by
HPLC
[1-‐5].
However,
each
of
these
techniques
possesses
regretful
weak-‐
ness
that
the
HPLC
method
was
untested
on
the
acidic
effect
for
the
mobile
phase.
Therefore
the
peaks
of
diosmin
and
rosmarinic
acid
which
were
completely
overlapped
have
not
been
separated.
Hence,
we
developed
a
new
method
for
simultaneous
determination
of
five
marker
compounds
(hesperidin,
rosmarinic
acid,
diosmin,
didymin,
and
buddleoside)
in
Menthae
Herba
by
HPLC/PDA.
Five
marker
components
were
separated
with
a
Phenomenex
Gemini
C18
(250
mm
×
4.6
mm,
5
µm)
column
by
one
step
gradient
elution
using
0.3%
trifluoroacetic
acid
in
methanol
(A)
and
water
(B)
as
the
mobile
phase.
The
flow
rate
was
1.0
mL/min,
and
the
UV
detector
wavelength
was
set
at
285
nm.
The
new
developed
method
was
successfully
validated
and
used
in
the
analysis
of
Menthae
Herba.
Validation
results
were
satisfactory
for
linearity
(r2>0.999),
recovery
(101.9~108.7%),
precision
(<0.21%),
accuracy
(100.50~111.00%),
stability
(98.47~101.01%)
and
robustness.
In
conclusion,
the
results
in-‐
dicated
that
the
established
HPLC/PDA
method
is
suitable
for
quantitation
for
quality
evalua-‐
tion
of
Menthae
Herba.
Acknowledgements:
This
work
was
supported
by
a
grant
from
the
National
Center
for
Standardization
of
Herbal
Medicine
funded
by
the
Ministry
of
Food
and
Drug
Safety,
Republic
of
Korea
(15172MFDS189).
Keywords: Menthae Herba, HPLC, drying condition, rosmarinic acid
References:
[1] Lim
HS,
Kim
JH,
Ha
HK,
Seo
CS,
Shin
HK.
Comparative
study
of
the
anti-‐inflammatory
effexts
of
Menthae
Herba
from
korea
and
China.
J
Kor
Pharmacogn
2012;
43:
231-‐238
[2] National
Pharmacopoeia
Commission
of
the
Peole’s
Republic
of
china.
Pharmacopoeia
of
the
People’s
Republic
of
China.
Beijing:
China
Medical
Sience
Press;
2010:
354
[3] Pharmacopeia
person
and
society.
Korean
Pharmacopeia.
Seoul:
Shinil
books;
2014:
1123.
[4] Shin
TY,
Kim
DK.
Antiallergic
activity
of
Menthae
Herba.
J
Kor
Pharmacogn
1998;
29:
248-‐
253
[5] Ye
D,
Zhao
M,
Shao
Y,
Ouyang
Z,
Peng
HS,
Han
BX,
Zhang
WW,
Gu
XM.
Relativity
of
commer-‐
cial
specification
of
Menthae
Herba
based
on
chemical
analysis.
Chin
J
Chin
Mater
Med
2015;
40:
215-‐217
P375
Pharmacognostical
studies
on
‘Mo-‐Si-‐Pool’
Marjahan
Acter
Kazi,
Hye-‐Jin
Kim,
Woo
Sung
Park,
Khan
Khalil
Atif
Ali,
Mi-‐Jeong
Ahn
College
of
Pharmacy
and
Research
Institute
of
Pharmaceutical
Sciences,
Gyeongsang
National
University,
Jinju
52828,
Korea
The
present
study
was
designed
to
establish
quality
control
parameters
for
pharmacognostic
evaluation
and
differentiation
of
seven
locally
occurring
Boehmeria
species
including
B.
nivea,
B.
nipononivea,
B.
pannosa,
B.
platanifolia,
B.
quelpaertensis,
B.
tricuspis,
B.
spicata,
and
one
variety
named
B.
tricuspis
var.
unicuspis
which
have
been
utilized
as
folk
medicine,
‘Mo-‐Si-‐
Pool’
in
Korea
[1].
Although
the
outer
morphological
study
of
these
species
had
been
report-‐
ed,
there
is
no
pharmacognostical
description
yet
[2].
Therefore,
inner
morphological
evalua-‐
tion
on
leaf
midrib,
petiole
and
stem
of
seven
Boehmeria
species
and
one
variety
was
accom-‐
plished,
and
preliminary
phytochemical
analysis
was
done
by
HPLC-‐DAD
profiling.
The
micro-‐
scopic
data
showed
discriminative
inner
morphological
characteristics
such
as
collenchyma
cell
layer,
diameter
of
parenchyma
cell
in
cortex,
thickness
of
cortex,
frequency
of
druse
and
diameter
of
vessels.
The
HPLC
profiles
exhibited
more
than
four
characteristic
peaks.
Among
these
peaks,
four
were
identified
as
flavonol
glycosides
and
a
phenanthroquinolizidine
alka-‐
loid
[3,
4],
and
the
content
of
these
components
in
Boehmeria
species
was
determined
by
HPLC-‐DAD.
These
findings
could
provide
the
scientific
criteria
for
the
proper
identification
and
estab-‐
lishment
of
standards
for
the
use
of
Boehmeria
species.
Acknowledgements:
This
research
was
financially
supported
by
the
Basic
Science
Research
Program
through
the
National
Research
Foundation
of
Korea
(NRF)
funded
by
the
Ministry
of
Science,
ICT
&
Fu-‐
ture
Planning
(NRF-‐2014R1A1A305070).
Keywords: Boehmeria species, inner morphological characteristics, HPLC profiles, alkaloids
References:
[1] Sancheti
S,
Sancheti
S,
Bafna
M,
Kim
H,
You
Y,
Seo
S.
Evaluation
of
antidiabetic,
antihyper-‐
lipidemic
and
antioxidant
effects
of
Boehmeria
nivea
root
extract
in
streptozotocin
in-‐
duced
diabetic
rats.
Rev
Bras
Farmacogn
2011;
21:
146-‐154
[2] Liang
SA.
A
phylogenetic
study
of
the
genus
Boehmeria
in
Korea
[dissertation].
Daegu:
Yeungnam
University,
2009
[3] Fiorentino
A,
Ricci
A,
D'Abrosca
B,
Golino
A,
Izzo
A,
Pascarella
MT,
Piccolella
S,
Esposito
A.
Kaempferol
glycosides
from
Lobularia
maritima
and
their
potential
role
in
plant
interac-‐
tions.
Chem
Biodivers
2009;
6:
204-‐217
[4] Cai
XF,
Jin
X,
Lee
DH,
Yang
YT,
Lee
K,
Hong
Y-‐S,
Lee
J-‐H,
Lee
JJ.
Phenanthroquinolizidine
al-‐
kaloids
from
the
roots
of
Boehmeria
pannosa
potently
inhibit
hypoxia-‐inducible
factor-‐1
in
AGS
human
gastric
cancer
cells.
J
Nat
Prod
2006;
69:
1095-‐1097
P376
A
prototype
traditional
Chinese
medicinal
plant
library
at
the
Na-‐
tional
Cancer
Institute
Min
He1,
Tanja
Grkovic2,
Christopher
C.
Thornburg2,
James
Whitt2,
Rhone
Akee2,
Jerell
Thompson2,
John
Britt2,
Libin
Jia3,
Jeffrey
White3,
David
Newman1,
Barry
O’Keefe1
1
Natural
Products
Branch,
Development
Therapeutic
Program,
Division
of
Cancer
Treatment
and
Diag-‐
nosis,
National
Cancer
Institute,
Frederick,
Maryland
21702,
2
Natural
Products
Support
Group,
Leidos
Biomedical
Research,
Inc.,
Frederick
National
Laboratory
for
Cancer
Research,
Frederick,
Maryland
21702,
3
Office
of
Cancer
Complementary
and
Alternative
Medicine,
Division
of
Cancer
Treatment
and
Diagnosis,
National
Cancer
Institute,
Rockville,
Maryland
20850
Traditional
Chinese
Medicine
(TCM)
has
evolved
over
thousands
of
years
in
China
and
other
Asian
countries
for
the
treatment
and
symptom
management
of
a
wide
range
of
medical
con-‐
ditions.
The
successful
development
of
anti-‐malaria
drug
artemisinin,
of
which
the
discovery
was
inspired
by
a
TCM
practice,
further
highlighted
the
importance
of
this
unique
resource
for
modern
drug
discovery
and
development.
Previously
a
prototype
TCM
library
has
been
estab-‐
lished
through
joint
efforts
of
US
and
Chinese
scientists,
consisting
of
more
than
200
authenti-‐
cated
medicinal
plant
and
fungal
species
that
collectively
represent
the
potential
therapeutic
content
of
the
majority
of
common
TCM
prescriptions
[1].
The
collection
has
duplicate
or
trip-‐
licate
kilogram
quantities
of
each
herb
species
that
were
collected
at
2–3
sites
with
precise
GPS
documentation
and
have
been
authenticated
visually
and
chemically.
A
processed
library
from
a
subset
of
this
collection
has
recently
been
developed
at
the
National
Cancer
Institute,
which
contains
both
the
organic
solvent
and
aqueous
extraction
of
331
samples
of
133
TCM
plant
species
in
96-‐
and
384-‐well
plate
formats
and
is
accessible
by
drug
discovery
research-‐
ers
worldwide.
Here
we
report
the
results
of
the
preliminary
“proof
of
principle”
assessment
of
the
anti-‐cancer
activity
of
this
crude
extract
library
in
NCI-‐60
human
cancer
cell
lines
screen,
as
well
as
the
chemical
profiling
and
comparison
analysis
of
selected
active
extracts.
It
is
hoped
that
this
prototype
TCM
plant
library
will
promote
scientific
discoveries
with
thera-‐
peutic
potential
and
enhance
international
efforts
to
systematically
evaluate
commonly
used
TCM
herbal
therapies.
Acknowledgements:
This
project
has
been
funded
in
whole
or
in
part
with
federal
funds
from
the
Nation-‐
al
Cancer
Institute,
National
Institutes
of
Health,
under
contract
HHSN261200800001E.
The
content
of
this
publication
does
not
necessarily
reflect
the
views
or
policies
of
the
Department
of
Health
and
Human
Services,
nor
does
mention
of
trade
names,
commercial
products,
or
organizations
imply
endorsement
by
the
U.S.
Keywords: Traditional Chinese Medicine, medicinal plants, HPLC-‐HRMS-‐SPE-‐NMR
References:
[1] Eisenberg
DM,
Harris
ES,
Littlefield
BA,
Cao
S,
Craycroft
JA,
Scholten
R,
Bayliss
P,
Fu
Y,
Wang
W,
Qiao
Y,
Zhao
Z,
Chen
H,
Liu
Y,
Kaptchuk
T,
Hahn
WC,
Wang
X,
Roberts
T,
Shamu
CE,
Clardy
J.
Developing
a
library
of
authenticated
Traditional
Chinese
Medicinal
(TCM)
plants
for
systematic
biological
evaluation-‐-‐rationale,
methods
and
preliminary
results
from
a
Sino-‐American
collaboration.
Fitoterapia
2011;
82:17-‐33
P377
Evaluation
of
Rumex
crispus
extracts
as
novel
matrix
metallopro-‐
teinase-‐1
(MMP-‐1)
inhibitors
Mine
Uzun,
L.
Omur
Demirezer
Hacettepe
University,
Faculty
of
Pharmacy,
Department
of
Pharmacognosy,
06100,
Sihhiye,
Ankara,
Tur-‐
key
Matrix
metalloproteinases
are
proteolytic
enzymes
responsible
for
breaking
down
extracellu-‐
lar
matrix
proteins
[1].
MMP-‐1
is
an
interstitial
collegenase
which
plays
crucial
roles
in
dis-‐
ease-‐related
physiological
and
pathological
processes
in
the
human
body.
Recent
studies
have
shown
that
inhibition
of
MMP-‐1
can
prevent
skin
degeneration
and
growth
or
spread
of
tu-‐
mours
in
various
cancers
[2,
3].
In
such
diseases,
inhibition
of
MMP
enzymes
could
be
a
part
of
treatment
strategy,
so
we
aim
to
inhibit
MMP-‐1
enzyme
by
Rumex
crispus
extracts
in
this
re-‐
search. Rumex
species
(family
Polygonaceae)
contain
anthranoids,
tannins,
flavonoids
and
naphthalene
skeletons
as
secondary
metabolites
and
have
various
biological
effects
[4].
In
this
research,
inhibitory
effects
of
Rumex
crispus
root,
leaf
and
fruit
extracts
on
MMP-‐1
enzyme
were
investigated
using
spectrophotometric
method.
Standardized
n-‐hexane,
dichloro-‐
methane,
ethyl
acetate,
ethanol
and
ethanol:
water
(70:30)
extracts
were
prepared
from
roots,
leaves
and
fruits.
Ac-‐PLG-‐(2-‐mercapto-‐4-‐methyl-‐pentanoyl)-‐LG-‐OC2H5
was
used
as
sub-‐
strate
and
NNGH
(N-‐Isobutyl-‐N-‐(4-‐methoxyphenylsulfonyl)
glycyl
hydroxamic
acid)
was
used
as
potent
MMP-‐1
inhibitor.
The
assays
were
performed
in
a
96-‐well
microplate.
Inhibitor
ac-‐
tivities
of
all
extracts
were
measured
in
100μg/mL,
400μg/mL,
and
800μg/mL
concentrations
at
412
nm.
The
aqueous
alcoholic
extracts
of
roots,
leaves
and
fruits
showed
the
highest
inhib-‐
itory
effect
on
MMP-‐1
enzyme
among
other
solvent
extracts
at
all
concentrations,
i.e.,
100
μg/mL,
400
μg/mL
and
800
μg/mL.
Inhibition
values
were
76%
for
root
extracts,
78%
for
leaf
extracts
and
83%
for
fruit
extracts,
while
NNGH
was
91%.
This
study
shows
that
aqueous
al-‐
coholic
extracts
of
Rumex
crispus
may
contain
potential
novel
MMP-‐1
inhibitors.
The
next
stage
of
our
study
will
be
to
develop
an
antiaging
formulation.
Acknowledgements:
This
study
was
supported
by
The
Scientific
and
Technological
Research
Council
of
Turkey
(TUBITAK)
(Project
No:
SBAG-‐214S147)
Keywords:
Matrix
metalloproteinase,
Rumex
crispus,
collegenase
inhibiton,
MMP-‐1
References:
[1]
Verma
RP,
Hansch
C.
Matrix
metalloproteinases
(MMPs):
chemical-‐biological
functions
and
(Q)
SARs.
Bioorgan
Med
Chem
2007;
15:
2223-‐2268
[2]
Yang
H,
Makaroff
K,
Paz
N,
Aitha
M,
Crowder
MW,
Tierney
DL.
Metal
ion
dependence
of
the
matrix
metalloproteinase-‐1
mechanism.
Biochemistry-‐US,
2015;
54:
3631-‐3639
[3]
Wojtoxicz-‐Praga
SM,
Dickson
RB,
Hawkins
MJ.
Matrix
metalloproteinase
inhibitors.
Invest
New
Drugs
1997;
15:
61-‐75
[4]
Vasas
A,
Gyapai
O,
Hohmann
J.
The
genus
Rumex:
review
of
traditional
uses,
phytochemis-‐
try
and
pharmacology.
J
Ethnopharmacol
2015;
175:
198-‐228
P378
α-‐Glucosidase
inhibitory
prenylated
anthranols
from
Harungana
madagascariensis
Oluwatosin
O.
Johnson1,2,
Ming
Zhao1,
Jordan
Gunn1,
Bernard
D.
Santarsiero1,
Zhi-‐Qi
Yin3,
Glo-‐
ria
A.
Ayoola2,
Herbert
A.
B.
Coker2,
Chun-‐Tao
Che1
1
Department
of
Medicinal
Chemistry
and
Pharmacognosy,
University
of
Illinois
at
Chicago,
Chicago,
IL
60612,
USA,
2
Department
of
Pharmaceutical
Chemistry,
University
of
Lagos,
CMUL
Campus,
Lagos
920001,
Nigeria,
3
Department
of
Natural
Medicinal
Chemistry
and
State
Key
Laboratory
of
Natural
Med-‐
icines,
China
Pharmaceutical
University,
Nanjing
210009,
China
Four
new
prenylated
anthranols,
harunganols
C−F
(1−4),
along
with
kenganthranol
A,
ha-‐
runganin,
and
ferruginin
A,
were
identified
from
the
leaves
of
Harungana
madagascariensis
Lam.
ex
Poir.
Harunganol
C
(1)
is
a
unique
symmetrical
anthranol
dimer
connected
via
a
methylene
group.
All
anthranols
were
evaluated
for
their
α-‐glucosidase
inhibitory
activities.
They
displayed
higher
potency
than
acarbose
except
for
3
and
4.
In
particular,
1
showed
an
IC50
value
of
1.2
μM.
Acknowledgements:
J.
Gunn
acknowledges
the
receipt
of
a
Ruth
Kirschstein
National
Research
Service
Award
(NRSA)
Pre-‐doctoral
Fellowship
(NIH
1F31AT00711701A1).
Keywords: Harungana madagascariensis, prenylated anthranols, harunganol, α-‐glucosidase
inhibitor
P379
Agathisflavone
reduces
inflammatory
modulators
in
microglia
BV2
cells
Mireia
Boluda
Navarro1,
Uchechukwu
Okorji1,
Ravikanth
Velagapudi1,
Priya
Jain1,
Mutalib
Aderogba2,
Olumayokun
A
Olajide1
1
Department
of
Pharmacy,
University
of
Huddersfield,
Huddersfield,
HD1
3DH,
United
Kingdom,
2
De-‐
partment of Chemistry, Obafemi Awolowo University, Ile-‐Ife, Nigeria
Agathisflavone (Figure 1) is a biflavonoid found in the leaves of Anacardium occidentale L.
[Anacardiaceae]. Diverse activities such as antineoplastic, hepatoprotective, antiviral proper-‐
ties [1] and antibacterial activities [2] have been previously reported for this secondary me-‐
tabolite. In this study, it is demonstrated that this compound also may possess anti-‐
inflammatory effects by reducing microglia cells activation. Cultured BV2 microglia cells
(ICLCATL03001, Banca Biologica e Cell Factory, Italy) were treated with Agathisflavone (5, 10
and 20 µM) 30 min before stimulation with LPS (100 ng/ml) derived from Salmonella typhi-‐
murium for a further 24 h. As a positive control cells treated only with LPS (100 ng/ml) were
used. Cell viability was assessed by MTT colorimetric assay. Supernatants were collected and
levels of nitric oxide (NO), tumor necrosis factor alpha (TNFα) and prostaglandine E2 (PGE2)
were measured using ELISA kits. Cyclooxygenase-‐2 (COX-‐2) and nitric oxide synthase (iNOS)
were determined by Western blotting. Agathisflavone produced significant reduction (p <
0.001) in the production of NO, TNFα and PGE2. Protein expressions of cyclooxygenase-‐2 and
iNOS were also abolished by agathisflavone pre-‐treatment (p < 0.001). Results show that
agathisflavone effectively exerts anti-‐inflammatory effects on BV2 cells microglia.
Figure
1.
Chemical
structure
of
agathisflavone
Acknowledgements:
Many
thanks
to
the
University
of
Huddersfield
for
awarding
me
with
a
fellowship
to
do
my
PhD.
Keywords:
Agathisflavone,
neuroinflammation,
BV2
microglia
References:
[1] André
N,
Lincopan
N,
Elida
I,
Díaz
C,
Fátima
J,
Midori
M,
Spira
B.
Experimental
and
Toxico-‐
logic
Pathology
Cytotoxicity
of
cashew
flavonoids
towards
malignant
cell
lines.
Exp
Toxi-‐
col
Pathol
2012;
64:
435-‐440
[2] Ajileye
OO,
Obuotor
EM,
Akinkunmi
EO,
Aderogba
MA.
Isolation
and
characterization
of
an-‐
tioxidant
and
antimicrobial
compounds
from
Anacardium
occidentale
L.
(Anacardiaceae)
leaf
extract.
J
King
Saud
Univ
Sci
2015;
27:
244-‐252
P380
A
new
sesquiterpene
from
Lebanese
wild
carrot
inhibits
prolifer-‐
ation
of
human
acute
myeloid
leukemia
cells
Joelle
Boulos1,
Ralph
Abi-‐Habib1,
Mirvat
El-‐Sibai1,
Wassim
Shebaby1,
Costantine
F.
Daher1,
Robin
I.
Taleb1,
Mohamad
A.
Mroueh2
1
School
of
Arts
and
Sciences,
Natural
Sciences
Department,
Lebanese
American
University,
PO
Box
36,
Byblos,
Lebanon,
2
School
of
Pharmacy,
Department
of
Pharmaceutical
Sciences,
Lebanese
American
Uni-‐
versity,
PO
Box
36,
Byblos,
Lebanon
Daucus
carota
L.
ssp.
carota
(Apiaceae),
also
known
as
wild
carrot,
is
a
commonly
used
medic-‐
inal
plant
in
folk
medicine
worldwide
for
the
treatment
of
several
ailments
including
cancer
[1].
Earlier
studies
in
our
labs
revealed
that
the
acetone:
methanol
oil
extract
of
the
plant
pos-‐
sesses
potent
anticancer
activity
against
a
panel
of
acute
myeloid
leukemia
(AML)
cells
[2].
The
aim
of
the
present
study
was
to
isolate
the
active
compound(s)
and
evaluate
its
anti-‐
cancer
activity.
Successive
column
chromatography
and
fractionation
led
to
the
isolation
of
a
new
sesquiterpene
unique
to
the
Lebanese
wild
carrot,
identified
as
1β-‐2-‐himachalen-‐6-‐ol
using
GC-‐MS
and
1D
and
2D
NMR
spectroscopy.
The
anticancer
activity
of
1β-‐2-‐himachalen-‐6-‐
ol
was
evaluated
against
two
AML
cell
line,
HL60
and
ML2.
Results
revealed
that
the
IC50
of
the
two
latter
cells
were
21
and
15
µg/mL,
corresponding
to
94.5
and
67.5
µM,
respectively.
Cell
cycle
analysis
using
flow
cytometry
revealed
an
increase
in
the
sub-‐G1
population.
Analy-‐
sis
of
cell
death
after
treatment
with
different
molecule
concentrations
showed
that
cells
were
positive
for
Annexin
V,
PI,
and
active
caspase
staining,
indicating
a
caspase
dependent
apop-‐
totic
cell
death.
In
conclusion,
1β-‐2-‐himachalen-‐6-‐ol
possesses
potent
anticancer
activity
against
the
tested
AML
cells,
an
effect
mediated
via
caspase
dependent
apoptotic
cell
death.
Hβ H
Hα Hβ
HO
Hβ Hα
Hα
Hα
H Hβ
H
Keywords:
Daucus
carota,
1β-‐2-‐himachalen-‐6-‐ol,
acute
myeloid
leukemia,
apoptosis,
caspase
References:
[1] Duke
AJ,
Bogenschutz-‐Godwin
MJ,
duCellier
J,
Duke
PK.
Handbook
of
Medicinal
Herbs.
2nd
edition.
Boca
Raton:
CRC
Press,
2002;
604
[2] Tawil
M,
Bekdash
A,
Mroueh
M,
Daher
CF,
Abi-‐Habib
RJ.
Wild
carrot
oil
extract
is
selectively
cytotoxic
to
human
acute
myeloid
leukemia
cells.
Asian
Pac
J
Cancer
Prev
2015;
16:761-‐
767
P381
Marantodes
pumilum
L.
plant
extracts
induce
apoptosis,
cell
cycle
arrest
and
inhibit
cell
migration
and
invasion
on
prostate
cancer
cell
lines.
Mohd
Mukrish
Mohd
Hanafi1,
Harisun
Yaakob2,
Mohamad
Roji
Sarmidi2,
Ramlan
Aziz2,
Jose
Maria
Prieto1
1
Department
of
Pharmaceutical
and
biological
chemistry,
UCL
School
of
Pharmacy,
29-‐39
Brunswick
Square,
WC1N
1AX,
London,
United
Kingdom
2
Institute
of
Bioproduct
Development
(IBD),
Universiti
Teknologi
Malaysia,
81310
UTM
Johor
Bahru,
Malaysia.
Prostate
cancer
ranks
ninth
overall
and
fourth
among
men
in
Malaysia
according
to
the
Na-‐
tional
Cancer
Registry.
The
crude
methanolic
extract
of
Marantodes
pumilum
L.
(also
known
as
Labisia
pumila
Benth.
&
Hook)
was
previously
identified
as
cytotoxic
against
prostate
can-‐
cer
cells
lines
[1].
This
study
aims
to
identify
for
the
first
time
the
active
fractions,
and
charac-‐
terize
mechanisms
of
action
against
PC3
prostate
cancer
cell
line.
The
results
showed
that
the
chloroform
and
n-‐hexane
fractions
of
the
plant
were
cytotoxic
towards
PC3
cell
line
with
low
IC50
values
(15-‐20
µg/mL).
Sulforhodamine
B
(SRB)
studies
indicate
that
the
active
fractions
showed
selective
cytotoxicity
only
to
PC3
cell
lines
when
tested
to
6
different
cells
lines
in-‐
cluding
HDFa
(normal
cell
line),
Mia-‐PaCa
II,
HepG2,
CaCo,
A549
and
SHSY5Y
(IC50
>200
µg/mL)
[2].
The
Annexin
V-‐FITC
study
indicated
that
cell
deaths
occur
via
apoptosis
through
significant
(p<0.01)
activation
of
caspases
3
and
7
[3].
This
finding
was
supported
by
the
sig-‐
nificant
decrease
(p<0.05)
in
the
mitochondrial
membrane
potential
(Δψ)
and
the
detection
of
nuclear
DNA
fragmentation
using
JC-‐1
MitoProbe™
and
TUNEL
assay
with
fluorescence
mi-‐
croscopy
[3].
Morphology
studies
with
DAPI
had
shown
the
characteristics
of
apoptotic
cells
including
cell
detachment,
cell
shrinkage
as
well
as
formation
of
apoptotic
bodies.
Cell
cycle
analysis
revealed
that
the
active
fractions
induced
cell
cycle
arrest
at
G2M
phase
[4].
The
2D
(Oris™
Cell
Migration)
and
3D
(modified
Boyden
chamber)
assays
showed
that
the
active
frac-‐
tions
significantly
reduced
(p<0.05)
both
PC3
cells
migration
and
invasion
respectively.
In
conclusion,
the
active
fractions
of
Marantodes
pumilum
L.
exhibit
antiproliferative
activity
through
the
induction
of
mitochondrial-‐driven
apoptosis
with
G2M
cell
arrest
and
inhibit
both
PC3
cells
migration
and
invasion
in
vitro.
Acknowledgements:
The
authors
would
like
to
acknowledge
the
Ministry
of
Education
Malaysia
and
Uni-‐
versiti
Teknologi
Malaysia
for
the
financial
assistance
under
the
Young
Academics
Training
Scheme
Keywords:
Prostate
cancer,
apoptosis,
migration,
invasion,
cell
cycle
arrest,
caspases
3
and
7,
DNA
fragmentation
References:
[1] Al-‐Mekhlafi
NA,
Shaari
K,
Abas
F,
Kneer
R,
Jeyaraj
EJ,
Stanslas
J.
Alkenylresorcinols
and
cytotoxic
activity
of
the
constituents
isolated
from
Labisia
pumila.
Phytochemistry
2012;
80:
42-‐49
[2] Vichai
V,
Kirtikara
K.
Sulforhodamine
B
colorimetric
assay
for
cytotoxicity
screening.
Nat
Protoc
2006;
1:
1112-‐1116
[3] He
Z,
Mangala
LS,
Theriot
CA,
Rohde
LH,
Wu
H,
Zhang
Y.
Cell
killing
and
radiosensitizing
effects
of
atorvastatin
in
PC3
prostate
cancer
cells.
J
Radiat
Res
2012;
53:
225-‐233
[4] Deep
G,
Singh
RP,
Agarwal
C,
Kroll
DJ,
Agarwal
R.
Silymarin
and
silibinin
cause
G1
and
G2–
M
cell
cycle
arrest
via
distinct
circuitries
in
human
prostate
cancer
PC3
cells:
a
compari-‐
son
of
flavanone
silibinin
with
flavanolignan
mixture
silymarin.
Oncogene
2006;
25:
1053-‐1069
P382
HPLC-‐DAD-‐MS
analysis
of
extracts
from
flowers,
leaves,
fruits
and
branches
of
Ligustrum
vulgare
and
their
effect
on
cytokines
se-‐
cretion
by
human
neutrophils
Monika E. Czerwińska, Małgorzata Warowny, Anna K. Kiss
Department
of
Pharmacognosy
and
Molecular
Basis
of
Phytotherapy,
Medical
University
of
Warsaw,
Banacha
1,
02-‐097
Warsaw,
Poland
Ligustrum
vulgare
L.
(Oleaceae)
is
a
shrub
commonly
found
in
Europe
and
Asia.
The
extracts
from
its
leaves
and
flowers
were
traditionally
used
in
oropharyngeal
inflammations,
aphthae
and
chronic
tonsillitis.
Fruits
served
as
mildly
laxative
agent
[1].
The
aim
of
the
study
was
a
comparison
of
the
composition
of
aqueous
and
ethanolic
extracts
from
leaves,
flowers,
fruits
and
branches
of
L.
vulgare
using
HPLC-‐DAD-‐MS/MS
method,
as
well
as
determination
of
total
phenolic
content
(TPC)
in
the
extracts.
In
order
to
compare
their
biological
activity
the
model
of
human
neutrophils,
which
are
cells
of
immune
system
involved
in
the
inflammation,
was
used.
The
effect
of
extracts
on
cytokines,
such
as
IL-‐8
and
TNFα,
secretion
by
neutrophils
was
determined.
HPLC-‐DAD-‐MS/MS
analysis
was
performed
on
a
Zorbax
SB
C18
column.
A
linear
gradient
sys-‐
tem
of
mobile
phase
A
(water/acetonitrile/formic
acid,
95:5:0.1,
v/v/v)
and
mobile
phase
B
(methanol)
was
used:
0-‐60
min.,
1-‐60%
B.
The
Folin-‐Ciocalteu
method
was
used
to
determine
TPC,
expressed
as
gallic
acid
equivalents.
The
effect
of
extracts
on
IL-‐8
and
TNFα
secretion
after
LPS-‐induced
stimulation
was
established
by
ELISA
assay.
The
most
abundant
compound
of
leaves,
flowers
and
branches
extracts
was
oleuropein,
whereas
the
major
compound
of
fruits
was
nuezhenide.
The
highest
TPC
was
observed
in
branches
ethanolic
(148.9
±
3.2
mg/g)
and
aqueous
(137.0
±
2.9
mg/g)
extracts.
These
ex-‐
tracts
were
also
the
most
effective
inhibitors
of
IL-‐8
and
TNFα
secretion.
The
branches
aque-‐
ous
extract
(25
µg/ml)
inhibited
IL-‐8
secretion
by
23.1
±
4.8
%,
whereas
ethanolic
extract
(100
µg/ml)
inhibited
TNFα
release
by
37.3
±
16.4
%
compared
with
control.
In
conclusion,
the
extracts
from
different
parts
of
L.
vulgare
constitute
a
potential
source
of
biologically
ac-‐
tive
compounds,
and
the
inhibition
of
cytokines
secretion
might
partially
justify
anti-‐
inflammatory
effectiveness
of
this
plant
material
preparations
in
the
treatment
of
disorders
in
folk
medicine.
Keywords: Ligustrum vulgare, oleuropein, secoiridoids, neutrophils
References:
[1] Fournier
P.
Encyclopédie
biologique,
Plantes
médicinales
et
vénéneuses
de
France.
Paris:
Paul
Lechevalier;
1948
P383
Chemical
composition
and
antimicrobial
activity
of
the
essential
oil
of
the
leaves
of
Cupressus
macrocarpa
Amal
M.
Saad1,
Magdy
M.
D.
Mohammed2,
Mosad
A.
Ghareeb1,
Wafaa
S.
Ahmed1,3,
Mohamed
A.
Farid4
1
Department
of
Medicinal
Chemistry,
Theodor
Bilharz
Research
Institute,
Kornish
El-‐Nile,
12661
Warrak
El-‐Hadar,
Giza,
Egypt,
2
Pharmacognosy
Department,
Pharmaceutical
and
Drug
Industries
Research
Divi-‐
sion,
National
Research
Centre
Dokki-‐12622,
Cairo,
Egypt,
3
Department
of
Chemistry,
College
of
Science
and
Arts,
Sajir,
Shaqra
University,
Kingdom
of
Saudi
Arabia,
4
Chemistry
of
Natural
and
Microbial
Prod-‐
ucts
Department,
Pharmaceutical
and
Drug
Industries
Research
Division,
National
Research
Centre,
Dok-‐
ki-‐12622,
Cairo,
Egypt
The
qualitative
(GC-‐MS)
and
quantitative
(GC-‐FID)
analyses
of
the
essential
oil
(EO)
constitu-‐
ents
of
the
leaves
of
Cupressus
macrocarpa
Hartweg.
ex
Gordon
(Cupressaceae)
revealed
the
presence
of
fifteen
identified
components
representing
86.29%
of
the
total
oil
composition,
which
were
identified
based
on
their
retention
times
and
mass
spectral
fragmentation
pat-‐
terns
to
be
α-‐terpineol
(19.01%),
camphenilone
(9.78%),
elemol
(8.92%),
2-‐tridecanone
(8.75%),
α-‐terpinyl
acetate
(8.62%),
2-‐pentyl-‐2-‐cyclopenten-‐1-‐one
(6.90%)
and
β-‐bisabolol
(5.83%)
as
the
major
components
[1,
2].
Furthermore,
the
antimicrobial
activity
of
the
EO
was
evaluated
by
agar
diffusion
method
[3]
against
eight
pathogenic
strains
including;
Pseudomo-‐
nas
aeruginosa,
Bacillus
subtilis,
Candida
albicans,
Fusarium
oxysporum,
Aspergillus
niger,
Fusarium
solani,
Escherichia
coli
and
Staphylococcus
aureus
at
dilution
(1:100
v/v)
with
inhibi-‐
tion
zones
ranged
from
10.5
to
21mm
in
comparison
to
Streptomycin.
In
conclusion,
the
EO
of
Cupressus
macrocarpa
leaves
exhibited
strong
antimicrobial
activity
against
certain
pathogen-‐
ic
microbial
strains;
therefore,
it
could
be
used
in
the
treatment
of
infectious
diseases.
Keywords:
Cupressus
macrocarpa,
Cupressaceae,
essential
oil,
GC-‐MS;
GC-‐FID,
antimicrobial
activity
References:
[1] Adams
RP.
Identification
of
essential
oils
by
Gas
Chromatography
Quadrupole
Mass
Spec-‐
trometry,
Allured
Publishing
Corporation,
Carol
Stream,
Illinois,
USA
2001
[2] Ibrahim
NA,
El-‐Seedi
HR,
Mohammed
MMD.
Constituents
and
biological
activity
of
the
chloroform
extract
and
essential
oil
of
Cupressus
sempervirens.
Chem
Nat
Compd
2009;
45:
309-‐313
[3] Ibrahim
NA,
El-‐Sakhawy
FS,
Mohammed
MMD,
Farid
MA,
Abdel-‐Wahed
NAM,
Deabes
DAH.
Chemical
composition,
antimicrobial
and
antifungal
activities
of
essential
oils
of
the
leaves
of
Aegle
marmelos
(L.)
Correa
growing
in
Egypt.
J
Appl
Pharm
Sci
2015;
5:
001-‐005
P384
The
Chemistry
of
African
Croton
species
Moses
K
Langat1,
Neil
Crouch2,3,
Beth
Ndunda4,
Jacob
O.
Midiwo4,
Areej
Aldhaher1,
Alaa
Alqahtani1,
Dulcie
A
Mulholland1,3
1Natural
Products
Research
Group,
Department
of
Chemistry,
FEPS,
University
of
Surrey,
Guildford,
GU2
7XH,
UK,
2Biodiversity
Research,
Monitoring
and
Assessment,
South
African
National
Biodiversity
Insti-‐
tute,
PO
Box
52099,
Berea
Road,
4007,
Durban,
South
Africa,
3School
of
Chemistry
and
Physics,
University
of
KwaZulu-‐Natal,
Durban,
4041,
South
Africa,
4Department
of
Chemistry,
University
of
Nairobi,
PO
Box
30197-‐00100,
Nairobi,
Kenya
The
genus
Croton
is
one
of
the
largest
of
Euphorbiaceae
sensu
stricto,
and
consists
of
over
1300
species
of
trees,
shrubs
and
herbs
that
are
distributed
worldwide
in
the
warm
tropics
and
subtropics.
It
is
reported
that
124
Croton
species
occur
in
continental
Africa
whilst
a
fur-‐
ther
156
species
are
endemic
to
Madagascar.
Another
12
species
occur
in
the
Indian
Ocean
islands
of
Comoros,
Mauritius,
Reunion
and
Sao
Tome
and
Principe
[1].
We
discuss
the
chem-‐
istry,
chemotaxonomic
patterns
and
biological
activities
of
selected
compounds
from
ten
Afri-‐
can
Croton
taxa:
C.
alienus,
C.
dichogamus,
C.
gratissimus
var.
gratissimus,
C.
megalobotrys,
C.
megalocarpoides,
C.
megalocarpus,
C.
menyhartii,
C.
pseudopulchellus,
C.
rivularis
and
C.
sylvati-‐
cus.
Examples
of
compounds
to
be
presented
include
cembranoids
(1-‐3)
from
C.
gratissimus
var.
gratissimus
[2,3],
ent-‐kauranes
(4-‐5)
from
C.
pseudopulchellus
[4],
ent-‐clerodanes
(6-‐8)
from
C.
sylvaticus,
C.
megalocarpus
and
C.
megalocarpoidies,
and
both
halimanes
(9)
and
croto-‐
folanes
(10-‐11)
from
C.
dichogamus.
Triterpenoids,
sesquiterpenoids,
flavonoids
and
cyclo-‐
hexanol
derivatives
from
Croton
will
also
be
discussed.
Selected
cembranoids
from
C.
gratis-‐
simus
were
tested
against
a
chloroquine-‐sensitive
strain
of
Plasmodium
falciparum
(D10)
and
against
the
PEO1
and
PEO1TaxR
ovarian
cancer
cell
lines
[2].
Compound
1
showed
moderate
activity
against
the
PEO1
(IC50
=
132
nM)
and
PEO1TaxR
(IC50
=
200
nM)
ovarian
cancer
cell
lines.
Selected
ent-‐kauranoids
were
tested
for
their
effects
on
Semliki
Forest
Virus
replication
and
for
cytotoxicity
against
human
liver
tumour
cells
(Huh-‐7
strain).
Other
Croton-‐derived
compounds
were
tested
for
antimicrobial
and
antifungal
activities
[5,6],
antiplasmodial
activi-‐
ty
using
two
strains
of
Plasmodium
falciparum,
antileishmanial
activities
against
Leishmania
donovanii
[5],
and
cytotoxic
activity
against
NCI59
cancer
cell
panels,
and
colorectal
and
VERO
cancer
cell
lines.
R1 HO H H
H R
OH HO O
H OH
O
H H H H H
O H O
CO2H CO2H
O O
1 4 5
2. R = OH R1 = H
3. R = H R1 = OH O
O O O O
O
H
H H O H
O
O O 10
O H O
O O O
O
H OH
H H O
CO2Me O HO O
H
O CO2Me
O 6 O
7 8 9
11
Acknowledgements:
University
of
Surrey,
University
of
KwaZulu
Natal
and
NRF-‐South
Africa
are
acknowledged
for
funding
Keywords: Croton, Euphorbiaceae, ent-‐clerodanes, cembranoids, crotofolanes
References
[1] WCSP
(2016).
'World
Checklist
of
Selected
Plant
Families.
Facilitated
by
the
Royal
Botanic
Gardens,
Kew.
Published
on
the
Internet;
http://apps.kew.org/wcsp/
Retrieved
on
12th
February
2016.'
[2] Mulholland
DA,
Langat
MK,
Crouch
NR,
Coley
HM,
Mutambi
EM,
Nuzillard
J-‐M.
Cem-‐
branolides
from
the
stem
bark
of
the
southern
African
medicinal
plant,
Croton
gratissimus
(Euphorbiaceae).
Phytochemistry
2010;
71:
1381-‐1386.
[3] Langat
MK,
Crouch
NR,
Smith
PJ,
Mulholland
DA.
Cembranolides
from
the
Leaves
of
Croton
gratissimus.
J
Nat
Prod
2011;
74:
2349-‐2355.
[4] Langat
MK,
Crouch
NR,
Pohjala
L,
Tammela
P,
Smith
PJ,
Mulholland
DA.
Ent-‐kauren-‐19-‐oic
acid
derivatives
from
the
stem
bark
of
Croton
pseudopulchellus
Pax.
Phytochem
Lett
2012;
5:
414
–
418.
[5] Ndunda
B,
Langat
MK,
Wanjohi
JM,
Midiwo
JO,
Kerubo
LO.
Alienusolin,
a
new
4α-‐
deoxyphorbol
ester
derivative,
and
crotonimide
C,
a
new
glutarimide
alkaloid
from
the
Kenyan
Croton
alienus.
Planta
Med
2013;
79:
1762-‐1766.
[6] Ndunda
B,
Langat
MK,
Midiwo
JO,
Omosa
LK.
Diterpenoid
Derivatives
of
Kenyan
Croton
sylvaticus.
Nat
Prod
Commun
2015;
10:
557-‐558.
P385
Phytochemical
and
biological
investigation
of
Calliandra
surina-‐
mensis
as
a
potential
treatment
for
diabetes
Abdullah
Alzahrani1,2,
Grainne
Abbott1,
Louise
C.
Young1,
John
Igoli1,3,
Alexander
I.
Gray1,
Va-‐
lerie
A
Ferro1
1Strathclyde
Institute
of
Pharmacy
and
Biomedical
Sciences,
University
of
Strathclyde,
161
Cathedral
Street,
G4
0RE,
Glasgow,
UK,
2Department
of
Pharmacology
and
Toxicology,
Faculty
of
Medicine,
Umm
Al-‐
Qura
University,
Makkah,
Saudi
Arabia,
3Phytochemistry
Research
Group,
University
of
Agriculture,
Ma-‐
kurdi,
P.M.B.
2373,
Benue
State,
Nigeria.
The
International
Diabetes
Federation
(IDF)
estimates
that
there
are
approximately
387
mil-‐
lion
people
with
diabetes
worldwide
[1].
The
current
pharmacological
management
for
diabe-‐
tes
has
been
reported
to
have
some
unpleasant
side
effects
such
as
hypoglycaemia,
lactic
aci-‐
dosis,
and
stomach
discomfort
[2].
Medicinal
plants
are
considered
a
good
source
for
mining
biologically
active
compounds.
The
current
study
investigates
the
potential
of
Calliandra
suri-‐
namensis
to
yield
anti-‐diabetic
compounds.
C.
surinamensis
leaves
and
bark
were
collected
from
the
Cayman
Islands
in
2015.
The
plant
materials
were
ground
and
then
Soxhlet
extrac-‐
tion
was
performed
using
hexane,
ethyl
acetate,
and
methanol.
Ethyl
acetate
and
hexane
ex-‐
tracts
were
subjected
to
open
column
silica
gel
chromatography,
while
the
methanol
extracts
were
separated
by
Sephadex
column
chromatography.
Structural
elucidation
of
isolated
com-‐
pounds
was
carried
out
using
Nuclear
Magnetic
Resonance
(1H
NMR
and
13C
NMR).
Three
compounds,
not
previously
reported
in
C.
surinamensis
were
isolated;
these
were
myricitrin,
lupeol,
and
ferulic
acid.
Myricitrin
is
known
to
possess
antioxidant
and
anti-‐inflammatory
ac-‐
tivity
[3],
lupeol
is
reported
to
have
anti-‐inflammatory
and
anti-‐cancer
activities
[4],
and
feru-‐
lic
acid
has
shown
anti-‐microbial
activities
in
previous
studies
[5].
C.
surinamensis
and
the
iso-‐
lated
compounds
have
not
been
investigated
for
anti-‐diabetic
activity
before
and
so
in
this
study,
crude
extracts
of
C.
surinamensis
were
biologically
tested
for
anti-‐diabetic
activity
using
various
in
vitro
assays
such
as
dipeptidyl
peptidase-‐IV
(DPPIV)
and
protein-‐tyrosine
phospha-‐
tase
1B
(PTP
1B)
enzyme
assays.
These
produced
negative
results
(P˃
0.05)
and
so
other
in
vitro
assays
(α-‐amylase,
and
α-‐glucosidase)
as
well
as
the
isolated
compounds
are
being
as-‐
sessed
for
anti-‐diabetic
potential.
Acknowledgements:
Abdullah
Alzahrani
is
a
PhD
student,
funded
by
the
Saudi
Arabian
Cultural
Bureau
in
London.
Keywords: Calliandria surinamensis, myricitrin, ferulic acid, lupeol, diabetes
References:
[1] International
Diabetes
Federation
(IDF).
Saudi
Arabia.
Available
at
http://www.idf.org/membership/mena/saudi-‐arabia.
Accessed
January
14,
2015
[2] National
Institute
for
Health
and
Clinical
Excellence
(NICE).
Type
2
diabetes
in
adults:
management.
Available
at
https://www.nice.org.uk/guidance/ng28.
Accessed
December
11,
2015
[3] Domitrovic
R,
Rashed
K,
Cvijanovic
O,
Vladimir-‐Knezevic
S,
Skoda
M,
Visnic
A.
Myricitrin
exhibits
antioxidant,
anti-‐inflammatory
and
antifibrotic
activity
in
carbon
tetrachloride-‐
intoxicated
mice.
Chem
Biol
Interact
2015;
230:
21-‐29
[4] Saleem
M.
Lupeol,
A
Novel
Anti-‐inflammatory
and
Anti-‐cancer
Dietary
Triterpene.
Cancer
Lett
2009;
285:
109-‐115
[5] Borges
A,
Ferreira
C,
Saavedra
MJ,
Simoes
M.
Antibacterial
activity
and
mode
of
action
of
ferulic
and
gallic
acids
against
pathogenic
bacteria.
Microb
Drug
Resist
2013;
19:
256-‐265
P386
The
effect
of
a
South
African
Helichrysum
sp.
against
important
pathogenic
mechanisms
of
Propionibacterium
acnes
M. N. de Canha1, N. Kishore1, N. Lall1
1 Department of Plant Science, University of Pretoria, Lynwood Road, 0083, Pretoria, South Africa
Acne
vulgaris
is
a
chronic
inflammatory
skin
disorder
affecting
the
pilosebaceous
follicles.
It
is
characterized
by
lesions
which
include
comedones,
papules
and
pustules.
In
more
severe
cas-‐
es
nodules
or
cysts
may
be
observed
[1].
Lesions
are
often
found
in
areas
rich
with
sebaceous
glands.
Although
acne
has
a
low
mortality
rate,
permanent
disfigurement
and
scarring
often
result
in
psychosocial
issues
such
as
depression
[2].
The
aim
of
this
study
was
to
investigate
the
potential
of
a
Helichrysum
sp.
as
a
natural
ingredient
in
anti-‐acne
agents.
The
Minimum
Inhibitory
Concentration
(MIC)
of
the
extract
was
determined
using
the
micro-‐dilution
broth
method.
The
cell
proliferation
kit
II
was
used
to
determine
the
effect
on
cell
viability.
Anti-‐
inflammatory
potential
against
COX
(cyclooxygenase)
II
was
determined
using
an
ELISA
kit.
Column
chromatographic
separation
was
performed
using
SiO2
(Silica
gel).
The
Helichrysum
sp.
extract
exhibited
an
MIC
of
7.81µg/ml.
Three
major
fractions
were
identified
Fg,
Fh
and
Fn.
The
MIC
for
each
fraction
was
observed
at
7.81µg/ml,
7.81µg/ml
and
62.50µg/ml,
respective-‐
ly.
The
extract
and
the
major
fraction
MICs
were
compared
with
known
antibiotic
tetracy-‐
cline,
with
an
MIC
of
1.56µg/ml.
Fraction
Fg
was
further
separated
using
a
basic
alumina
col-‐
umn.
The
cytotoxicity
of
the
extract
was
tested
using
human
keratinocytes
(HaCaT)
and
ex-‐
hibited
a
fifty
percent
inhibitory
concentration
(IC50)
of
201.30
±
23.45µg/ml.
The
Selectivity
Index
(SI)
was
calculated
as
25.77
(IC50/MIC).
The
SI
of
the
extract
shows
a
worthy
therapeu-‐
tic
window
(>10)
indicating
the
potential
for
inhibition
of
inflammatory
cytokines
such
as
IL-‐
8
and
TNF-‐α
[3].
The
extract
inhibited
human
COX
II
activity
with
an
IC50
of
21.97
±
4.27µg/ml.
The
potential
of
this
Helichrysum
sp.
as
a
natural
ingredient
for
acne
has
been
shown.
More
research
needs
to
be
conducted
to
strengthen
the
cosmetic
application
showing
the
need
for
clinical
trial
research
for
efficacy
and
safety.
Acknowledgements:
University
of
Pretoria
and
the
National
Research
Foundation
(NRF)
Keywords: Acne vulgaris, Helichrysum sp., anti-‐inflammatory, cytotoxicity
References:
[1] Dessinioti
C,
Katsambas
AD.
The
role
of
Propionibacterium
acnes
in
acne
pathogenesis:
facts
and
controversies.
Clin
Dermatol
2010;
28:
2-‐7.
[2] Moggadam
MR,
Ardabili
NS,
Maleki
N,
Soflaee
M.
Correlation
between
the
severity
and
type
of
acne
lesions
with
serum
zinc
levels
in
patients
with
acne
vulgaris.
BioMed
Res
Int
2014;
2014:
1-‐3
[3] Fang
W,
Peng
F,
Yi
T,
Zhang
C,
Wan
C,
Xu
H,
Lam
CW,
Yang
X.
Biological
activity
and
safety
of
Tripterygium
extract
prepared
by
sodium
carbonate
extraction.
Molecules
2012;
17:
11113-‐11123
P387
α-‐amylase
and
α-‐glucosidase
inhibitor
activities
of
secondary
metabolites
from
Arcytophyllum
thymifolium
M.
De
Leo1,
A.
Braca
1,
M.
B.
Vera
Saltos
1,2,
I.
Faraone3,
N.
Malafronte4,
L.
Milella3,
N.
De
Tom-‐
masi4
1
Dipartimento
di
Farmacia,
Università
di
Pisa,
Pisa,
Via
Bonanno
33,
56126
Pisa,
Italy,
2
Departamento
de
Ciencias
de
la
Vida,
Universidad
de
las
Fuerzas
Armadas,
ESPE,
v.
General
Rumiñahui
s/n,
Sangolqui,
Ecuador,
3
Dipartimento
di
Scienze,
Università
degli
Studi
della
Basilicata,
Viale
dell’Ateneo
Lucano
10,
85100
Potenza,
Italy,
4
Dipartimento
di
Farmacia,
Università
di
Salerno,
Via
Giovanni
Paolo
II,
84084
Fisciano,
Salerno,
Italy
Plants
belonging
to
Arcytophyllum
genus
are
widespread
in
Central
and
South
America
mainly
in
Costa
Rica,
Panama,
Venezuela,
Colombia,
Ecuador,
Peru
and
Bolivia
[1].
Aerial
parts
infu-‐
sion
and/or
decoction
are
used
in
traditional
medicine
for
the
treatment
of
colic
and
indiges-‐
tion
[2].
Only
one
study
was
previously
reported
in
the
literature,
showing
the
presence
of
flavonoids
and
triterpenoids
from
Arcytophyllum
genus
[3].
A.
thymifolium
(Ruiz
&
PAV)
Standl
(Rubiaceae)
is
a
shrub
growing
in
Ecuador
in
the
typical
Ande
mountain
ecosystem
[1].
In
the
present
study,
a
phytochemical
investigation
of
A.
thymifolium
aerial
parts
was
per-‐
formed
for
the
first
time.
The
dried
and
powdered
plant
material
was
sequentially
extracted
with
n-‐hexane,
CHCl3,
CHCl3-‐MeOH
(9:1)
and
MeOH.
The
CHCl3
extract
was
first
subjected
to
a
flash
chromatography
through
Biotage
Isolera™,
and
fractions
obtained
were
successively
eluted
on
RP-‐HPLC
and
HPCPC.
The
MeOH
extract
was
partitioned
between
n-‐BuOH
and
H2O
and
the
n-‐BuOH
fraction
was
preliminary
chromatographed
on
Sephadex
LH-‐20
column,
sub-‐
sequently,
fractions
obtained
were
submitted
to
RP-‐HPLC.
Twenty-‐three
compounds
(8
cou-‐
marins,
4
flavonoids,
9
iridoids,
2
caffeic
acid
derivatives),
including
five
new
secondary
me-‐
tabolites,
were
finally
isolated
and
identified
by
NMR
and
MS
analyses.
The
hypoglycaemic
properties
of
known
and
new
compounds
were
also
evaluated
measuring
extracts
and
pure
compounds
α-‐amylase
and
α-‐glucosidase
inhibitory
effects,
by
using
acarbose
as
positive
con-‐
trol
[4].
The
iridoid
asperulosidic
acid
showed
the
higher
activity
as
α
-‐amylase
inhibitor,
hav-‐
ing
an
IC50
of
70
µM,
moderately
higher
than
acarbose
(IC50
26.3
µM),
while
the
new
flavanone
7-‐O-‐(3-‐metylbut-‐2-‐enyl)oxy
eriodictyol
resulted
to
be
the
most
active
as
α-‐glucosidase
inhibi-‐
tor
with
an
IC50
of
30
µM,
sensibly
lower
than
acarbose
(IC50
402.7
µM).
A
molecular
docking
study
is
in
progress
to
gain
information
on
α-‐glucosidase-‐7-‐O-‐(3-‐metylbut-‐2-‐enyl)
oxy
eri-‐
odictyol
interaction.
Keywords:
Arcytophyllum
thymifolium,
Rubiaceae,
iridoids,
coumarins,
α-‐amylase
inhibitors,
α-‐glucosidase
inhibitors
References:
[1] Mena
PV.
A
revision
of
the
Genus
Arcytophyllum
(Rubiaceae:
Hedyotideae).
New
York
Bo-‐
tanical
Garden,
New
York;
1990
[2] Monigatti
M,
Bussmann
RV,
Weckerle
CS.
Medicinal
plant
use
in
two
Andean
communities
located
at
different
altitudes
in
the
Bolívar
Province,
Peru.
J
Ethnopharmacol
2013;
145:
450-‐464
[3] De
Feo
V,
Della
Valle
C,
De
Simone
F,
Pizza
C.
Chemical
constituents
and
antibacterial
activ-‐
ity
of
Arcytophyllum
nitidum
H.B.K.
Annali
di
Chimica
1992;
82:
149-‐159
[4] Vera
Saltos
MB,
Naranjo
Puente
BF,
Faraone
I,
Milella
L,
De
Tommasi
N,
Braca
A.
Inhibitors
of
α-‐amylase
and
α-‐glucosidase
from
Andromachia
igniaria
Humb.
&
Bonpl.
Phytochem
Lett
2015;
14:
45-‐50
P388
Chrysophanol-‐
and
nepodin-‐8-‐O-‐β-‐D-‐glucopyranoside
from
Ru-‐
mex
acetosella,
the
cytotoxicity
towards
drug
sensitive
and
multi-‐
drug
resistant
T
leukaemia
cancer
cells
Nadire
Ozenver1,2,
Mohamed
Saeed2,
Zuhal
Guvenalp3,
L.
Omur
Demirezer1,
Thomas
Efferth2
1Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Hacettepe
University,
Sihhiye,
06100,
Ankara,
Turkey,
2Department
of
Pharmaceutical
Biology,
Institute
of
Pharmacy
and
Biochemistry,
Johannes
Gu-‐
tenberg
University,
Staudinger
Weg
5,
55128
Mainz,
Germany,
3Ataturk
University,
Faculty
of
Pharmacy,
Department
of
Pharmacognosy,
25240,
Erzurum,
Turkey
Rumex
acetosella
L.
(Polygonaceae)
is
traditionally
used
in
cancer
treatment
by
few
tribes
in
Canada
[1].
It
contains
anthraquinones
[2],
flavonoids
[2,
3]
and
other
phenolics
[4].
We
isola-‐
ted
anthraquinone
and
naphtalene
glycosides
by
using
different
column
chromatography
techniques
and
determined
their
structures
with
NMR
spectroscopy
as
chrysophanol-‐8-‐O-‐β-‐D
glucopyranoside
and
nepodin-‐8-‐O-‐β-‐D-‐glucopyranoside.
Anthraquinones
inhibit
tumor
growth
on
some
cancer
cells.
Additionally
naphtalene
derivatives
are
shown
to
inhibit
tumor
growth
at
nM
ranges
on
few
cell
lines.
Multidrug
resistance
in
cancer
represents
a
major
prob-‐
lem
in
chemotherapy.
Natural
compounds
having
fewer
side
effects
can
be
proper
candidates
for
cancer
treatment.
Therefore
potential
drug
candidates
from
natural
sources
need
to
be
searched.
In
this
study
we
assessed
the
cytotoxicity
of
an
anthraquinone
and
a
naphthalene
derivative
from
the
roots
of
R.
acetosella
against
drug
sensitive
CCRF-‐CEM
and
multidrug
re-‐
sistant
P-‐
glycoprotein-‐over-‐expressing
CEM/ADR5000
leukemia
cells.
The
resazurin
assay
was
used
to
determine
the
cytotoxicity
of
the
compounds.
The
recorded
IC50
values
for
chry-‐
sophanol-‐8-‐O-‐
β-‐D-‐glucopyranoside
and
nepodin-‐8-‐O-‐
β-‐D-‐glucopyranoside
ranged
from
69.05
μM
to
74.9
μM
(towards
leukemia
CCRF-‐CEM
cells)
and
from
104.52
μM
to
82.9
μM
(towards
leukemia
CEM/ADR5000
cells),
compared
to
established
chemotherapeutic
agent
“doxorubicin”
which
showed
0.0007
μM
(towards
leukemia
CCRF-‐CEM
cells)
and
48.9
μM
to-‐
wards
leukemia
CEM/ADR5000
cells.
Even
if
the
compounds
didn’t
show
remarkable
cytotox-‐
icity
on
leukemia
cells,
further
studies
need
to
be
conducted
on
natural
compounds
that
could
have
a
potential
to
combat
with
drug
resistance
in
cancer.
Acknowledgments:
This
study
was
supported
by
The
Scientific
and
Technological
Research
Council
of
Turkey
(TÜBITAK)
2214-‐A
scholarship.
Keywords: Anthraquinone, naphtalene, cytotoxicity, Resazurin reduction assay, mutidrug
resistantance (MDR), cancer
References:
[1] Leonard SS, Keil D, Mehlman T, Proper S, Shi XL, Harris GK. Essiac tea: Scavenging of
reactive oxygen species and effects on DNA damage. J Ethnopharmacol 2006; 103: 288-‐
296
[2] Duke JA. Handbook of Biological Active Phytochemicals & Their Activity: CRC-‐Press; 1992.
[3] Thomas R. The Essiac Report: The True Story of a Canadian Herbal Cancer Remedy and of
the Thousands of Lives it Continues to Save: Alternative Treatment Information Network;
1993
[4]
Đurđević
L,
Gajić
G,
Jarić
S,
Kostić
O,
Mitrović
M,
Pavlović
P.
Analysis
of
benzoic
and
cinnamic
acid
derivatives
of
some
medicinal
plants
in
Serbia.
Arch
Biol
Sci
2013;
65:
603-‐
609
P389
Cytotoxicity
of
main
anthraquinone
aglycons
towards
drug
sensi-‐
tive
and
multi
drug
resistant
T
leukaemia
cancer
cells
Nadire
Ozenver1,2,
Mohamed
Saeed2,
L.
Omur
Demirezer1,
Thomas
Efferth2
1Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Hacettepe
University,
Sihhiye,
06100,
Ankara,
Turkey,
2Department
of
Pharmaceutical
Biology,
Institute
of
Pharmacy
and
Biochemistry,
Johannes
Gu-‐
tenberg
University,
Staudinger
Weg
5,
55128
Mainz,
Germany
Cancer
as
one
of
the
leading
causes
of
mortality
is
increasing
regularly
worldwide
[1].
Multi-‐
drug
resistant
(MDR)
cancer
cells
constitute
a
serious
problem
in
chemotherapy.
The
modula-‐
tion
of
resistance-‐mediated
genes
or
proteins
by
phytochemicals
can
be
an
attractive
strategy
to
overcome
MDR
resistance
in
cancer,
in
regards
to
the
diversity
of
chemical
structures
in
plant
kingdom.
Naturally
occurring
anthranoids
as
anthracene
derivatives
are
mostly
present
in
Rumex,
Rhamnus,
Rheum
species
[2,
3].
Anthraquinones,
the
major
group
of
naturally
oc-‐
curring
quinones
are
distinguished
by
a
large
structural
diversity,
wide
range
of
biological
activity,
and
low
toxicity
[4].
They
exhibit
a
wide
range
of
bioactivities
such
as
anticancer
as
well
as
antimicrobial,
hypotensive,
antimalarial,
analgesic
and
anti-‐inflammatory
[4-‐6].
In
this
study
we
investigated
the
cytotoxicity
of
the
four
anthraquinones
emodin,
aloe-‐emodin,
physcion
and
rhein
against
drug
sensitive
leukemia
CCRF-‐CEM
cells
and
their
resistance
coun-‐
terpart
P-‐glycoprotein-‐over-‐expressing
CEM/ADR5000
leukaemia
cells.
The
resazurin
reduc-‐
tion
assay
was
used
to
evaluate
the
cytotoxicity
of
the
compounds.
The
recorded
IC50
values
for
compounds
were
ranged
from
11.95
μM
to
142.44
μM
towards
CCRF-‐CEM
and
from
22.41
μM
to
90.69
μM
towards
CEM/ADR5000
cells.
Clinically
used
anticancer
drug
doxorubicin
had
the
following
IC50
values:
0.0007
μM
towards
CCRF-‐CEM
and
48.9
μM
towards
CEM/ADR5000
cells.
Compared
to
their
corresponding
sensitive
cell
lines,
collateral
sensitivity
(hypersensi-‐
tivity)
was
observed
in
drug-‐resistant
P-‐glycoprotein
over-‐expressing
CEM/ADR5000
cells
(0.67
fold)
to
physcion.
In
conclusion,
we
demonstrated
the
cytotoxicity
of
few
main
anthra-‐
quinone
aglycons
against
drug-‐sensitive
and
MDR
leukaemia
cancer
cells.
In
particular
aloe-‐
emodin
might
be
a
potential
cytotoxic
natural
agent
that
deserves
more
investigations
to
de-‐
velop
novel
antineoplastic
drugs.
Acknowledgments:
This
study
was
supported
by
The
Scientific
and
Technological
Research
Council
of
Turkey
(TÜBITAK)
2214-‐A
scholarship.
Keywords:
Anthraquinone,
cytotoxicity,
Resazurin
reduction
assay,
mutidrug
resistant
(MDR),
cancer
References:
[1]
Prakash
O,
Kumar
A,
Kumar
P,
Ajeet
A.
Anticancer
potential
of
plants
and
natural
products:
A
review.
Am
J
Pharm
Sci
2013;
1:
104-‐115
[2]
Demirezer
LÖ,
Kuruüzüm
A.
A
comparative
chemotaxonomic
study
on
eleven
Rumex
spe-‐
cies
growing
in
Turkey.
FABAD
1997;
22:
153-‐158
[3]
Dictionary
of
Natural
Products
(DNP)
in
CD-‐ROM
(1999)
Version
8:1,
Chapman
&
Hall,
Boca
Raton,
Floride,
USA
[4]
Eyong
KO,
Kuete
V,
Efferth
T.
10
-‐
Quinones
and
Benzophenones
from
the
Medicinal
Plants
of
Africa.
In,
Medicinal
Plant
Research
in
Africa.
Oxford:
Elsevier;
2013;
351-‐391
[5]
Dave
H,
Ledwani
L.
A
review
on
anthraquinones
isolated
from
Cassia
species
and
their
applications.
Indian
J
Nat
Prod
Resour
2012;
3:
291-‐319
[6]
Demirezer
LO,
Kuruüzüm-‐Uz
A,
Bergere
I,
Schiewe
HJ,
Zeeck
A.
The
structures
of
antioxidant
and
cytotoxic
agents
from
natural
source:
anthraquinones
and
tannins
from
roots
of
Ru-‐
mex
patientia.
Phytochemistry
2001;
58:
1213-‐1217
P390
Comparative antimicrobial and antioxidant studies of two closely
related species of Saba
Nana Ama Mireku-‐Gyimah1, Kofi Annan2, Ji-‐Kai Liu3, Kwame Sarpong1
1Department of Pharmacognosy, 2Department of Herbal Medicine, Faculty of Pharmacy and Pharmaceu-‐
tical
Sciences,
College
of
Health
Sciences,
Kwame
Nkrumah
University
Science
and
Technology,
Kumasi,
Ghana,
3State
Key
Laboratory
of
Phytochemistry
and
Plant
resources
in
West
China,
Chinese
Academy
of
Sciences,
Kunming
Institute
of
Botany,
Kunming,
China
Saba
senegalensis
(A.
DC)
Pichon
and
Saba
thompsonii
(A.
Chev)
Pichon
(Apocynaceae)
are
closely
related
species
of
the
genus
Saba,
native
to
West
Africa.
Although
widely
used
in
Gha-‐
na
for
wounds,
dysenteric
diarrhoea
and
tuberculosis
[1],
there
is
no
known
scientific
verifi-‐
cation
of
the
use
of
these
climbers.
Close
resemblance
of
the
plants
often
results
in
them
being
used
interchangeably
during
traditional
preparations.
The
study
investigates
antimicrobial
and
antioxidant
activities
of
their
crude
alcoholic
extracts
for
scientific
credence
and
the
effect
of
using
the
plants
interchangeably.
Minimum
Inhibitory
Concentration
(MIC)
of
Saba
senega-‐
lensis
(SS)
and
Saba
thompsonii
(ST)
were
obtained
using
micro-‐dilution
method
[2]
at
con-‐
centrations
30−0.0146
mg/mL
screened
against
selected
microorganisms
(Gram-‐positive:
Staphylococcus
aureus,
Streptococcus
pyogenes,
Gram-‐negative:
Pseudomonas
aeruginosa,
Escherichia
coli,
Salmonella
typhi,
fungi:
Candida
albicans).
Antioxidant
effects
were
assayed
using
DPPH
radical
scavenging
[3],
total
antioxidant
capacity
[4]
and
total
phenolic
content
[5].
Both
extracts
had
activity
against
one
or
more
of
selected
microorganisms
(MICs
ranging
from
7.5
mg/mL–30
mg/mL).
EC50
values
of
0.02325
mg/mL
and
0.01931
mg/mL
were
rec-‐
orded
for
DPPH
radical
scavenging
activity
of
SS
and
ST,
respectively
(reference
drug,
vitamin
E,
0.00567
mg/mL).
Total
antioxidant
capacities
of
SS
and
ST
were
264.8
±
31.50
mg/g
and
276.3
±
42.75
mg/g
of
vitamin
E,
respectively.
Total
phenolic
content
expressed
per
gram
equivalent
of
tannic
acid
was
determined
as
109.1
±
2.24
mg/g
for
SS
and
87.33
±
2.43
mg/g
for
ST.
SS
has
broad
spectrum
activity
and
higher
phenolic
content
whereas
ST
has
a
narrow
spectrum,
higher
total
antioxidant
capacity
and
DPPH
radical
scavenging
activity.
Traditional
uses
of
S.
senegalensis
and
S.
thompsonii
as
antimicrobial
agents
are
valid
and
interchanging
them
in
preparations
may
result
in
varied
treatment
outcomes.
Acknowledgements:
OWSD
(Organisation
for
Women
in
Science
for
the
Developing
World)
and
Sida
(Swedish
International
Development
Cooperation
Agency)
for
sponsorship
Keywords:
Saba,
traditional
use,
comparative
studies,
antimicrobial,
antioxidant
References:
[1] Burkill
HM.
The
useful
plants
of
west
tropical
Africa,
1995;
3:
168-‐169
[2] Wiegand
I,
Hilpert
K,
Hancock
RE.
Agar
and
broth
dilution
methods
to
determine
the
minimal
inhibitory
concentration
(MIC)
of
antimicrobial
substances.
Nat
Protoc
2008;
3:
163-‐175
[3] Govindappa
M,
Sadananda
T,
Channabasava
R,
Raghavendra
VB.
In
vitro
anti-‐
inflammatory,
lipoxygenase,
xanthine
oxidase
and
acetycholinesterase
inhibitory
activity
of
Tecoma
stans
(L.)
Juss.
Ex
kunth.
Int
J
Pharma
Bio
Sci
2011;
2:
275-‐285
[4] Prieto
P,
Pineda
M,
Aguilar
M.
Spectrophotometric
quantitation
of
antioxidant
capacity
through
the
formation
of
a
phosphomolybdenum
complex:
specific
application
to
the
determination
of
vitamin
E.
Anal
Biochem
1999;
269:
337-‐341
[5] McDonald
S,
Prenzler
PD,
Antolovich
M,
Robards
K.
Phenolic
content
and
antioxidant
activity
of
olive
extracts.
Food
Chem
2001;
73:
73-‐84
P391
Study
of
Pterocarpus
erinaceus,
a
promising
plant
from
Togo
to
treat
infectious
diseases
Nassifatou
Koko
Tittikpina1,2,3,5,
Frédéric
Nana2,3,
Stéphane
Fontanay2,3,4,
Komlan
Batawila5,
Koffi
Akpagana5,
Gilbert
Kirsch2,3,
Raphaël
E.
Duval2,3,4,
Patrick
Chaimbault2,3,
Claus
Jacob1
1
Faculty
of
Pharmacy,
Department
of
Bio-‐organic
Chemistry,
Building
B
2.1.,
Room
1.13.
Saarland
State
University,
Campus
D-‐66123
Saarbrücken,
Germany,
2
CNRS,
UMR
7565,
SRSMC,
F-‐54506
Vandœuvre-‐lès-‐
Nancy,
France,
3
Université
de
Lorraine,
UMR
7565,
SRSMC,
F-‐54506
Vandœuvre-‐lès-‐Nancy,
France,
4
ABC
Platform®,
F-‐54001
Nancy,
France,
5
Laboratoire
de
Botanique
et
Ecologie
Végétale,
Université
de
Lomé,
BP
1515,
Lomé,
Togo
Ethnobotanical
surveys
run
in
Togo
have
come
out
with
Pterocarpus
erinaceus
as
the
most
used
plant
by
traditional
healers
to
treat
various
types
of
infectious
diseases
[1-‐3].
A
bio-‐
guided
screening
has
been
proposed
to
study
it.
Extraction
and
fractionation
processes
have
been
carried
out
on
the
different
plant
parts:
leaves,
barks
and
roots.
The
extracts
obtained
have
shown
good
activities
on
a
wide
range
of
Gram
positive
bacteria:
Enterococcus
faecalis
ABC
3
(ATCC
29212),
Staphylococcus
aureus
ABC
1
(ATCC
29213);
as
well
as
on
Gram
nega-‐
tive
bacteria:
Escherichia
coli
ABC
5
(ATCC
25922),
Klebsiella
pneumoniae
ABC
42,
Enterobac-‐
ter
cloacae
ABC
291,
Pseudomonas
aeruginosa
ABC
4
(ATCC
27853)
and
Acinetobacter
bau-‐
mannii
ABC
14
[4].
The
extracts
presented
a
very
good
activity
with
a
Minimum
Inhibitory
Concentration
(MIC)
ranging
from
32
µg/ml
to
256
µg/ml
especially
on
S.
aureus
(MIC:
64
µg/ml).
Extracts
have
also
shown
a
noticeable
activity
on
the
Methicillin-‐Resistant
S.
aureus
(MRSA)
with
a
MIC
at
64
µg/ml.
The
same
MIC
is
obtained
on
Staphyloccoccus
epidermidis.
Interestingly,
is
it
noted
the
conservation
of
the
activities
with
fractions
coming
from
raw
ex-‐
tracts
and
the
absence
of
toxicity
to
normal
cells
lines
namely
MRC5
(ATCC
CCL-‐171)
cells
[5].
Analytical
studies
(HPLC,
GC-‐MS,
LC-‐MS
and
NMR)
are
presently
being
performed
on
the
most
active
fractions
to
identify
the
active
principles
[6-‐8].
Even
if
the
analytical
studies
are
still
going
on,
some
compounds
have
already
been
isolated
and/or
identified.
Some
of
them
were
already
found
in
other
plants
but,
to
the
best
of
our
knowledge,
never
reported
in
P.
erinaceus.
Among
these
compounds,
we
found
some
phytosterols
(e.g
stigmasterol,
β-‐sistosterol
and
campestrol),
pentacyclic
triterpenoids
(α-‐
and
β-‐amyrin),
(poly)
phenolic
compounds
and
fatty
acids.
The
next
step
will
to
be
to
evaluate
the
effect
of
the
new
and/or
existing
isolated
compounds
on
the
same
biological
targets.
Acknowledgements:
We
are
thankful
to
the
Schlumberger
Foundation
through
its
Faculty
For
the
Future
Program
for
funding
our
PhD
studies
at
both
the
University
of
Saarland
and
the
University
of
Lorraine
and
the
‘GradUS
Global’
of
the
University
of
Saarland
for
complementary
funding.
We
acknowledge
tech-‐
nical
help
from
Stéphanie
PHILIPPOT.
Keywords: Togo, Pterocarpus erinaceus Poir, plant extracts, antibacterial activities
References:
[1] Tittikpina
NK.
Contribution
à
l’évaluation
des
Propriétés
Anti-‐Microbiennes
de:
Pterocar-‐
pus
Erinaceus
Poir
(Faboïdeae),
Daniellia
oliveri
(Rolfe)
Hutch.
et
Dalz
(Caesalpinoïdeae)
et
Anchomanes
difformis
(Blume)
Engler
(Araceae),
utilisées
en
médecine
traditionnelle
dans
la
Préfecture
de
Tchamba
(TOGO)
[PharmD
Dissertation].
Lomé
(Togo)
:
Université
de
Lomé
(University
of
Lomé),
2012
[2] Tittikpina
NK,
Agban
A,
Gbogbo
KA,
Houkou
YP,
Pereki
H,
Batawila
K,
Akpagana
K.
Évalua-‐
tion
des
propriétés
antimicrobiennes
de
Pterocarpus
erinaceus
Poir
(Faboïdeae)
et
Dan-‐
iellia
oliveri
(Rolfe)
Hutch.
et
Dalz
(Caesalpinoïdeae),
utilisées
en
médecine
traditionnelle
au
Togo.
Int
J
Biol
Chem
Sci
2013;
7:1586-‐1594
[3] Tittikpina
NK,
Ejike
ECCC,
Castelluci
Estevam
E,
Nasim
MJ,
Griffin
S,
Chaimbault
P,
Kirsch
G,
Atakpama
W,
Batawila
K,
Jacob
C.
Togo
to
go:
Products
and
compounds
derived
from
local
plants
for
the
treatment
of
diseases
endemic
in
Sub-‐Saharan
Africa.
Afr
J
Tradit
Comple-‐
ment
Altern
Med
2016;
13:
85-‐94
[4] CLSI.
Methods
for
dilution
antimicrobial
susceptibility
tests
for
bacteria
that
grow
aerobi-‐
cally;
Approved
Standard,
8th
ed.
CLSI
Document
M07-‐A8.
Wayne,
PA:
Clinical
and
Labor-‐
atory
Standards
Institute;
2009
[5] Mosmann
T.
Rapid
colorimetric
assay
for
cellular
growth
and
survival:
application
to
pro-‐
liferation
and
cytotoxicity
assays.
J
Immunol
Methods
1983;
65:
55-‐63
[6] Czepukojc
B,
Baltes
AK,
Kelkel
M,
Cerella
C,
Viswanathan
UM,
Salm
F,
Burkholz
T,
Schneider
C,
Montenarh
M,
Diederich
M,
Jacob
C.
Synthetic
polysulfane
derivatives
induce
cell
cycle
arrest
and
apoptotic
cell
death
in
human
hematopoietic
cancer
cells.
Food
Chem
Toxicol
2014;
64:
249-‐257
[7] Eddaya
T,
Boughdad
A,
Sibille
E,
Chaimbault
P,
Zaid
A,
Amechrouq
A.
Biological
activity
of
Sapindus
mukorossi
Gaerten
(Sapindaceae)
aqueous
extract
against
Thysanoplusia
orichal-‐
cea
(Lepidoptera:
Noctuidae).
Ind
Crops
Prod
2013;
50:
325-‐332
[8] Kim
CS,
Subedi
L,
Kim
SY,
Choi
SU,
Kim
KH,
Lee
KR.
Diterpenes
from
the
trunk
of
Abies
holo-‐
phylla
and
their
potential
neuroprotective
and
anti-‐inflammatory
activities.
J
Nat
Prod
2016;
79:
387-‐394
P392
Cholinesterase
inhibitory
potentials
of
some
Turkish
medicinal
plants
Nehir
Unver
Somer,
Buket
Bozkurt
,
Ceren
Emir,
Gulen
Irem
Kaya,
Ahmet
Emir,
Mustafa
Ali
Onur
Department of Pharmacognosy, Faculty of Pharmacy, Ege University, 35100 Bornova, Izmir, Turkey
Alzheimer’s
disease
(AD)
is
a
complex
neurodegenerative
process
characterized
by
memory
impairment
and
cognitive
dysfunction.
Currently,
acetylcholinesterase
(AChE)
inhibitors
are
the
main
therapeutic
approach
for
the
treatment
of
AD.
In
healthy
brains,
AChE
hydrolyzes
the
majority
of
acetylcholine,
while
butyrlcholinesterase
(BChE),
another
enzyme
involved
in
the
degradation
of
acetylcholine,
plays
a
secondary
role.
As
AD
progresses,
BChE
activity
sig-‐
nificantly
increases.
Therefore,
both
enzymes
are
considered
as
important
targets
in
the
treatment
of
AD
[1].
Many
natural
products
have
been
shown
to
have
anti-‐cholinesterase
ac-‐
tivity
[2].
Among
these,
galanthamine,
an
Amaryllidaceae
alkaloid,
is
used
in
AD
therapy
[1,
2].
In
this
work,
we
have
determined
the
cholinesterase
inhibitory
potentials
of
some
plant
spe-‐
cies
used
in
Turkish
folk
medicine
[3].
The
investigated
species
were
Rumex
crispus
L.,
Stachys
cretica
subsp.
lesbiaca
Rech.
fil.,
Dianthus
calocephalus
Boiss.,
Asperula
tenuifolia
Boiss.,
Equi-‐
setum
ramosissimum
Desf.,
Vinca
herbacea
Waldst.
et
Kit.,
Salvia
sclarea
L.,
Scrophularia
canina
L.
and
Salvia
pinnata
L.
MeOH
extracts
of
the
aerial
parts
of
these
plants
were
screened
for
anti-‐cholinesterase
activity
against
the
enzymes
AChE
and
BChE
using
a
microplate
assay
modified
from
in
vitro
Ellman’s
method
[4]
at
10,
100
and
1000
µg/ml
concentrations
(final
concentrations
in
the
assay
2.5,
25
and
250
µg/ml).
Galanthamine
was
used
as
a
positive
con-‐
trol.
Among
the
extracts
screened,
MeOH
extract
of
Rumex
crispus,
exhibited
the
highest
inhi-‐
bition
against
AChE
(91.6
%)
at
250
µg/ml,
whereas
the
other
extracts
had
below
50
%
inhibi-‐
tion
on
this
enzyme.
Moreover,
remarkable
butyrylcholinesterase
inhibition
(ranging
from
63.88-‐99
%),
was
observed
for
Rumex
crispus,
Stachys
cretica
subsp.
lesbiaca,
Asperula
tenuifo-‐
lia,
Equisetum
ramosissimum,
Vinca
herbacea,
Salvia
sclarea,
and
Salvia
pinnata
extracts
at
250
µg/ml.
Acknowledgements:
This
study
was
financially
supported
by
Ege
University
Research
Fund
(09/ECZ/021).
Keywords: Anticholinesterase activity, medicinal plants
References:
[1] Lane
RM,
Potkin
SG,
Enz
A.
Targeting
acetylcholinesterase
and
butyrylcholinesterase
in
dementia.
Int
J
Neuropsychopharmacol
2006;
9:
101−124
[2] Williams
P,
Sorribas
A,
Howes
MJR.
Natural
products
as
a
source
of
Alzheimer’s
drug
leads.
Nat
Prod
Rep
2011;
28:
48−77
[3] Baytop
T.
Therapy
with
plants
in
Turkey.
Istanbul:
Nobel
Tıp
Press,2nd
ed,
1999
[4] Ellman
GL,
Courtney
KD,
Andres
V,
Featherstone
RM.
New
and
rapid
colorimetric
determi-‐
nation
of
acetylcholinesterase
activity.
Biochem
Pharmacol
1961;
7:
88−95
P393
Phenolic
compounds
of
Phlomis
samia
L.
from
Turkey
Göger
Fatih1,
Köse
Yavuz
Bülent2,
Kırımer
Neşe1
1
Anadolu
University,
Faculty
of
Pharmacy,
Department
of
Pharmacognosy,
26470,
Eskisehir,
Turkey,
2
Anadolu
University,
Faculty
of
Pharmacy,
Department
of
Pharmaceutical
Botany,
26470,
Eskisehir,
Tur-‐
key
The
genus
Phlomis
L.
from
Lamiaceae
is
populated
from
south
western
part
of
Asia,
Europe
to
Africa,
which
is
represented
in
the
world
approximately
by
113
taxa,
where
58
grows
in
Tur-‐
key
[1,
2].
Some
preparations
of
these
taxa
are
used
in
folk
medicine
for
their
analgesic,
diu-‐
retic,
tonic,
anti-‐diarrheic
preparations
and
to
treat
various
conditions
such
as
gastric
ulcer,
diabetes,
inflammation,
hemorrhoids,
wounds,
etc.
[3].
This
present
study
aimed
to
identify
and
characterize
the
phytochemical
content
of
Phlomis
samia
L.
methanol
extract
for
the
phe-‐
nolic
compounds.
The
phenolic
compounds
were
determined
by
a
HPLC
system
coupled
to
a
LC-‐
MS/MS
instrument
where
the
separations
were
performed
on
a
ODS
octadecyl
silica
gel
analytical
column.
As
a
result,
forsythoside
B
(3.3%),
5-‐caffeoylquinic
acid
(0.6%),
verbasco-‐
side
(3.6%),
and
luteolin
glucoside
(0.02%)
were
determined
as
the
major
constituents
of
the
P.
samia
methanol
extract
to
the
best
of
our
knowledge
for
the
first
time.
Phytochemical
and
in
vitro
biological
evaluations
of
the
plant
material
are
ongoing
for
discovery
of
new
applica-‐
tions.
Keywords: Phlomis samia, phenolic compounds, LC-‐MS/MS
References:
[1]
Li
MX,
Shang
XF,
Jia
ZP,
Zhang
RX.
Phytochemical
and
biological
studies
of
plants
from
the
genus
Phlomis.
Chem
Biodivers
2010;
7:
283-‐301
[2]
Dadandi,
MY.
Türkiye’nin
Phlomis
L.
(Lamiaceae)
Cinsi
Revizyonu:
Doctoral
Thesis,
Gazi
university,
Ankara
Turkey
2002
[3]
Baytop
T.
Therapy
with
Medicinal
Plants
in
Turkey,
Past
and
Present:
Nobel
Tıp
Publica-‐
tions,
Istanbul,
Turkey
1999:
193
P394
Investigation
of
polysaccharide
composition
in
medicinal
and
non-‐medicinal
aloes
Louise
I.
Ahl1,
Henriette
L.
Pedersen2,
William
G.
T.
Willats2,
Nina
Rønsted1,
Olwen
M.
Grace3
1
Natural
History
Museum
of
Denmark,
Faculty
of
Science,
University
of
Copenhagen,
Sølvgade
83S,
DK-‐
1307
Copenhagen
K,
Denmark,
2
Plant
and
Environmental
Sciences,
Faculty
of
Science,
University
of
Co-‐
penhagen,
Thorvaldsensvej
40,
DK-‐1871
Frederiksberg
C,
Copenhagen,
3
Royal
Botanic
Gardens,
Kew,
Surrey
TW9
3AE,
United
Kingdom
The
genus
Aloe
comprises
over
500
species
of
leaf
succulents
found
throughout
Africa,
Mada-‐
gascar
and
the
Arabian
Peninsula.
Aloe
vera
is
a
particularly
widely
known
species,
and
is
used
as
an
ingredient
in
products
from
herbal
medicine
to
cosmetics
and
household
commod-‐
ities.
Aloe
vera
dominates
the
market
worldwide,
but
as
many
as
25%
of
the
Aloe
species
are
used
locally,
a
few
of
them
even
supported
by
small
industries
[1,2].
The
major
chemical
com-‐
ponents
of
the
tissue
used
in
these
products
are
polysaccharides
-‐
highly
complex
molecules
found
in
the
spongy
leaf
mesophyll
(referred
to
as
Aloe
gel).
Studies
of
other
plants
used
in
traditional
medicine
have
identified
bioactive
polysaccharides
and
shown
them
to
have
im-‐
mune-‐modulatory
activity
[3].
In
Aloe,
polysaccharides
extracted
from
the
leaf
mesophyll
have
been
associated
with
both
anti-‐inflammatory
and
immune-‐modulatory
activity
[4].
As
poly-‐
saccharides
are
highly
complex,
they
are
often
studied
indirectly
using
monosaccharide
anal-‐
yses.
A
recent
study
showed
that
the
monosaccharide
composition
of
31
species
of
Aloe
is
conservative,
with
90%
of
the
variation
accounted
for
glucose,
mannose
and
xylose
as
the
ma-‐
jor
constituents
[5].
The
extent
to
which
leaf
mesophyll
polysaccharides
differ
between
Aloe
species
has,
until
now,
remained
unclear.
Here,
we
present
data
from
an
initial
study
of
the
polysaccharide
composition
of
11
species
of
Aloe,
using
carbohydrate
microarrays
and
23
monoclonal
antibodies
binding
to
epitopes
representing
six
distinct
types
of
plant
polysaccha-‐
rides
[6,7].
Marked
variation
in
the
binding
patterns
were
observed
between
the
studied
spe-‐
cies,
indicating
potential
diversity
of
polysaccharides
within
the
genus
Aloe
not
observed
in
analyses
of
the
monosaccharides.
Acknowledgements:
This
work
was
supported
by
the
Villum
Foundation
as
part
of
the
PLANET
project:
Understanding
plant
evolution
and
diversity
in
a
changing
world.
Martin
Aarseth-‐Hansen
and
Paul
Rees
are
acknowledged
for
assistance
with
the
collection
of
plant
material,
and
Paul
Knox
for
the
monoclonal
antibodies
Keywords:
Aloe,
microarrays,
polysaccharides,
medicinal
plant
use,
gel
References:
[1] Grace
OM,
Simmonds
MSJ,
Smith
MF,
van
Wyk
AE.
Documented
utility
and
biocultural
value
of
Aloe
L.
(Asphodelaceae):
A
review.
Economic
Bot
2009;
63:
167-‐178
[2] Grace
OM.
Current
perspectives
on
the
economic
botany
of
the
genus
Aloe
L.
(Xanthor-‐
rhoeaceae).
S
Afr
J
Bot
2011;
77:
980-‐987
[3] Paulsen
BS,
Barsett
H.
Bioactive
pectic
polysaccharides.
Adv
Polym
Sci
2005;
186:
69-‐101
[4] Steenkamp
V,
Stewart
MJ.
Medicinal
applications
and
toxicological
activities
of
Aloe
prod-‐
ucts.
Pharm
Biol
2007;
45:
411-‐420
[5] Grace
OM,
Dzajic
A,
Jäger
AK,
Nyberg
NT,
Önder
A,
Rønsted
N.
Monosaccharide
analysis
of
succulent
leaf
tissue
in
Aloe.
Phytochemistry
2013;
93:
79-‐87
[6] Moller
I,
Sørensen
I,
Bernal
AJ,
Blaukopf
C,
Lee
K,
Øbro
J,
Pettolino
F,
Roberts
A,
Mikkelsen
JD,
Knox
JP,
Bacic
A,
Willats
WGT.
High-‐throughput
mapping
of
cell-‐wall
polymers
within
and
between
plants
using
novel
microarrays.
The
Plant
Journal
2007;
50:
1118-‐1128
[7] Fagel
JU,
Pedersen
HL,
Melgosa
SV,
Ahl
LI,
Salmean
AA,
Egelund
J,
Rydahl
MG,
Clausen
MH,
Willats
WGT.
Carbohydrate
microarrays
in
plant
science.
Methods
Mol
Biol
2012;
918:
351-‐362
P395
Spectral
and
chemical
studies
on
Magnoflorine
from
Zanthoxylum
armatum
DC
(Rutaceae)
leaves
and
evaluation
of
its
antistress
potential
Nitin
Verma1,
R.
L.
Khosa2
1School
of
Pharmacy
and
Emerging
Science
(SPES),
Baddi
University
of
Emerging
Science
and
Technolo-‐
gy,
Makhunmajara,
Baddi,
Distt-‐Solan,
H.
P.
173205,
India.
2School
of
Pharmacy,
Bharat
Institute
of
Technology,
Partapur
Bypass,
NH#58,
Meerut,
250002,
India
The
purification
of
the
chloroform
extract
from
the
leaves
of
Zanthoxylum
armatum
DC
(Ru-‐
taceae)
using
column
chromatography
furnished
Magnoflorine,
a
1,2,10,11-‐tetrasubstituted
aporphine
alkaloid.
The
structure
of
the
compound
was
determined
through
spectral
data
analysis
(UV-‐spectroscopy,
FTIR
spectroscopy,
1H
NMR
spectroscopy
and
mass
spectrometry)
and
chemical
means.
The
anti-‐stress
effect
of
compound
was
evaluated
by
Open-‐Field
behav-‐
ior
test,
Y-‐maze
test,
swimming
endurance
test,
autoanalgesia,
traction
test,
rotating
rod
test,
effect
on
pentobarbitone
sleeping
time,
cold
restraint
stress
induced
ulceration,
adrenal,
brain,
liver
and
spleen
weight
following
cold
restraint
stress,
corticosterone
levels
following
cold
restraint
stress,
monoamine
oxidase
levels
following
cold
restraint
stress
and
lipid
pe-‐
roxidation
levels
following
cold
restraint
stress.
The
compound
showed
a
potent
anti-‐stress
activity.
The
results
are
in
agreement
with
the
normalizing
affects
of
the
said
compound
in
various
stress
models.
The
details
of
protocols
and
procedures
will
be
discussed
during
presentation.
Keywords: Zanthoxylum armatum, Magnoflorine, aporphine alkaloid, anti-‐stress
References:
[1] Rudzik
AD,
Hester
JB,
Jang
AH,
Stray
RN,
Friss
W.
The
Benzodiazepines.
Ed:
Garattini
S.
Mussinii
E.
and
Randall
L.O.
Raveen
Press
New
York,
1973,
pp.
285
[2] Brodie
DA,
Hanson
HM.
A
study
of
the
factors
involved
in
the
production
of
gastric
ulcers
by
the
restraint
technique.
Gastroenterol
1960;
38:
353-‐360.
[3] Ohkawa
H,
Ohishi
N,
Yagi
K.
Assay
for
lipid
peroxides
in
animal
tissues
by
thiobarbituric
acid
reaction.
Anal
Biochem
1979;
95:
351-‐358.
P396
Inhibition
of
interleukin-‐8
release
in
human
neutrophils
by
Melodorum
fruticosum
Nora
Engels1,
Birgit
Waltenberger,
Barbara
Michalak2,
Loi
Huynh3,
Hung
Tran3,
Anna
Kiss2,
Hermann
Stuppner1
1Institute
of
Pharmacy/
Pharmacognosy
and
Center
for
Molecular
Biosciences,
University
of
Innsbruck,
Innrain
80-‐82,
6020
Innsbruck,
Austria,
2Department
of
Pharmacognosy
and
Molecular
Basis
of
Phyto-‐
therapy,
Faculty
of
Pharmacy,
Medical
University
of
Warsaw,
02-‐097
Warsaw,
Poland,
3Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
University
of
Medicine
and
Pharmacy,
41-‐43
Đinh
Tiên
Hoàng,
Bến
Nghé,
Quận
1,
Hồ
Chí
Minh,
Vietnam
The
treatment
of
inflammatory
disorders
in
Western
medicine
relies
heavily
on
the
use
of
non-‐steroidal
anti-‐inflammatory
drugs
(NSAIDs)
and
corticosteroids
[1].
However,
currently
available
treatment
options
are
often
unsatisfactory.
In
the
course
of
this
project
we
investi-‐
gated
plants
used
in
Vietnamese
traditional
medicine
for
the
treatment
of
inflammatory
dis-‐
orders.
Dried
plant
material
from
Aleurites
moluccana
(folium
and
caulis),
Alstonia
scholaris
(folium
and
caulis),
Clinacanthus
nutans
(folium),
Ficus
benghalensis
(folium),
and
Melodorum
fruticosum
(folium)
was
successively
extracted
with
dichloromethane
(DCM)
followed
by
methanol.
Extracts
were
tested
for
their
inhibitory
activity
on
lipopolysaccharide
(LPS)-‐
triggered
interleukin-‐8
(IL-‐8)
expression
in
human
neutrophils.
At
a
concentration
of
10
µg/mL,
the
Melodorum
fruticosum
DCM
extract
showed
highly
potent
activity
with
88
%
inhi-‐
bition
of
IL-‐8
release.
Standard
flow
cytometric
probe
using
propidium
iodide
staining
[2]
re-‐
vealed
that
the
active
Melodorum
fruticosum
DCM
extract
also
induced
apoptosis.
Bioactivity
guided
fractionation
of
the
DCM
extract
monitored
by
the
IL-‐8
assay
led
to
the
isolation
of
melodamide
A
[3],
which
significantly
inhibited
the
expression/production
of
IL-‐8
by
stimu-‐
lated
neutrophils
at
a
concentration
range
of
2.5-‐25
µM
without
showing
cytotoxic
effects
on
the
tested
cells.
Further
phytochemical
analysis
resulted
in
the
isolation
and
identification
of
several
additional
compounds
including
N-‐trans-‐cinnamoyltyramine,
alpinetin,
tsugafolin,
5,7-‐dimethoxy-‐4'-‐hydroxyflavanone,
2',4'-‐dihydroxy-‐4,6'-‐dimethoxydihydrochalcone,
2′,4′-‐
dihydroxy-‐4,6′-‐dimethoxychalcone,
and
2′,6′-‐dihydroxy-‐4′-‐methoxychalcone
from
the
DCM
extract,
and
5-‐methoxy-‐naringenin,
catechin,
kaempferol-‐3-‐O-‐α-‐L-‐rhamnoside,
and
quercetin-‐
3-‐O-‐α-‐L-‐rhamnoside
from
the
ethanol
extract.
The
pharmacological
activity
of
melodamide
A
may
justify
the
traditional
use
of
Melodorum
fruticosum
to
treat
inflammatory
disorders.
Keywords:
Vietnamese
traditional
medicine,
inflammation,
neutrophils,
Melodorum
fruti-‐
cosum,
melodamide
A
References:
[1] Wang
Q,
Kuang
H,
Su
Y,
Sun
Y,
Feng
J,
Guo
R,
Chan
K.
Naturally
derived
anti-‐inflammatory
compounds
from
Chinese
medicinal
plants.
J
Ethnopharmacol
2013;
146:
9-‐39
[2] Schinella
G,
Aquila
S,
Dade
M,
Giner
R,
Recio
MdC,
Spegazzini
E,
de
Buschiazzo
P,
Tournier
H,
Rios
JL.
Anti-‐inflammatory
and
apoptotic
activities
of
pomolic
acid
isolated
from
Cecropia
pachystachya.
Planta
Med
2008;
74:
215-‐220
[3] Chan
H-‐H,
Hwang
T-‐L,
Thang
TD,
Leu
Y-‐L,
Kuo
P-‐C,
Nguyet
BTM,
Dai
DN,
Wu
T-‐S.
Isolation
and
synthesis
of
melodamide
A,
a
new
anti-‐inflammatory
phenolic
amide
from
the
leaves
of
Melodorum
fruticosum.
Planta
Med
2013;
79:
288-‐294
P397
Isomer
of
bergenin
and
phytosterol
from
the
stem
bark
of
Mallo-‐
tus
leucodermis
Aiza Syuhada Mohd Yusoff1, Norizan Ahmat1,2, Humera Naz2
1 Faculty of Applied Sciences, Universiti Teknologi MARA, 40450 Shah Alam, Selangor, Malaysia, 2 Atta Ur
Rahman
Institute
for
Natural
Products
Discovery,
Universiti
Teknologi
MARA,
UiTM
Puncak
Alam,
42300
Bandar
Puncak
Alam,
Selangor,
Malaysia
Mallotus
is
one
of
the
most
diverse
and
richest
genera
of
the
Euphorbiaceae
family
[1],
and
consists
of
approximately
150
species
distributed
in
tropical
and
sub-‐tropical
regions
of
Asia
[2].
Mallotus
species
are
well
known
to
possess
medicinal
properties
and
has
been
reported
to
be
rich
in
flavonoids,
terpenoids,
phenolic
compounds,
and
phloroglucinol
derivatives
[3].
Compounds
and
extracts
from
Mallotus
species
exhibited
antioxidant,
antimicrobial,
antiviral,
anti-‐inflammatory
and
cytotoxic
activities
[3].
Mallotus
leucodermis
Hook
f.
is
a
mid-‐canopy
tree
that
can
be
found
easily
in
Peninsular
Malaysia,
Borneo,
Sumatra
and
Thailand
[4].
Mallo-‐
tus
leucodermis
is
commonly
known
as
“balik
angin
bopeng”
in
Malaysia,
and
is
used
to
treat
skin
complaints
[5].
A
phytochemical
study
was
conducted
on
the
stem
bark
of
M.
leu-‐
codermis
collected
from
National
Park
Kuala
Keniam,
Pahang,
Malaysia.
The
dried
powder
of
the
stem
bark
(1.3kg)
was
macerated
with
acetone
to
yield
66.0g
of
crude
extract.
The
crude
extract
was
fractionated
using
vacuum
liquid
chromatography
to
give
six
fractions.
Fraction
5
was
further
purified
using
radial
chromatography
to
afford
an
isomer
of
bergenin
(1)
(23.0mg).
Purification
of
fraction
1
using
radial
chromatography
gave
a
mixture
of
stigmas-‐
terol
and
β-‐sitosterol
(2.9mg).
The
structure
of
compounds
was
elucidated
by
spectroscopic
methods
including
1D
and
2D
NMR,
UV-‐Vis,
FTIR,
MS
and
by
comparison
with
literature
data.
Acknowledgements:
Faculty
of
Applied
Sciences,
UiTM
Shah
Alam
is
acknowledged
for
the
provision
of
facilities
and
technical
assistance.
Keywords: Euphorbiaceae, Mallotus, bergenin, stigmasterol, sitosterol
References:
[1] Thakur
HA,
Patil
DA.
Taxonomic
and
phylogenetic
assessment
of
the
Euphorbiaceae:
A
Review.
J
Expl
Sci
2011;
2:
37-‐
46
[2] Schatz
GE.
Royal
Botanic
gardens,
Kew
and
Missouri
Botanical
Gardens,
Saint-‐Louis
2001;
503
[3] Riviere
C,
Hong
VNT,
Hong
QT,
Chataigne
G,
Hoai
NN,
Dejaeger
B,
Tistaert
C,
Kim
TNT,
Heyden
YV,
Van
MC
and
Quetin-‐leclercq
J.
Mallotus
species
from
Viatnamese
mountain-‐
ous
areas:
phytochemistry
and
pharmacology
activities.
Phytochem
Rev
2010;
9:
217-‐
253
[4] Bollendorff
SM,
Van
Welzen
PC
and
Slik
JWF.
A
taxonomic
revision
of
Mallotus
section
Pol-‐
yadenii
(Euphorbiaceae).
Blumea
2000;
45:
319-‐340
[5] Faridah
H,
Nurulhuda
H.
The
Use
of
Medicinal
Plant
Species
by
the
Temuan
Tribe
of
Ayer
Hitam
Forest,
Selangor,
Peninsular
Malaysia,
J
Trop
Agric
Sci
1999;
22:
85-‐94
P398
Flavones
from
the
twigs
of
Cynometra
cauliflora
Linn.
Nik Fatini Nik Azmin1,2, Norizan Ahmat1,2
1 Atta-‐ur-‐Rahman Institute for Natural Product Discovery, Universiti Teknologi MARA (UiTM), Puncak
Alam
,
42300
Bandar
Puncak
Alam,
Selangor
Darul
Ehsan,
2
Faculty
of
Applied
Sciences,
Universiti
Teknologi
MARA
(UiTM),
Shah
Alam,
40450
Selangor
Darul
Ehsan
Cynometra
cauliflora
L.
is
a
member
of
the
Fabaceae
and
has
a
vernacular
name
nam-‐nam.
It
is
believed
to
be
native
of
Malaysia
and
is
cultivated
in
Indonesia
and
India
[1].
C.
cauliflora
is
a
small,
much-‐branched
perennial
tree
growing
to
5
m
tall.
It
is
a
typical
underutilized
fruit
tree
that
is
used
as
a
traditional
medicine
in
treating
several
diseases.
The
tree
is
also
cultivated
as
an
ornamental
[2].
A
decoction
of
the
leaves
is
traditionally
used
for
treating
diabetes
and
hy-‐
perlipidemia
[3,
4],
the
fruits
are
used
for
curing
loss
of
appetite,
and
the
seed
oil
is
used
to
cure
skin
diseases
[5].
Previous
work
on
anti
and
pro-‐lipase
activities
have
led
to
the
isolation
of
kaempferol-‐3-‐O-‐rhamnoside
as
an
active
constituent
from
the
plant
leaves
[4].
Other
stud-‐
ies
reported
the
antiacetylcholinesterase,
antityrosinase,
antioxidant
and
α-‐glucosidase
inhib-‐
itory
activities
of
leaf
extract
[6].
This
study
focused
on
investigating
the
constituents
from
the
twigs
of
this
plant
using
conventional
method.
Dried
twigs
of
Cynometra
cauliflora
L.
were
macerated
in
acetone
to
obtain
crude
extract
and
fractionated
using
vacuum
liquid
chroma-‐
tography
(VLC).
The
isolation
and
purification
of
the
chosen
fractions
by
a
combination
of
multiple
radial
and
preparative
thin
layer
(pTLC)
chromatographic
techniques
then
afforded
four
pure
compounds,
three
flavones:
luteolin
(1),
acacetin
(2),
3’,4’,7-‐trihydroxyflavone
(3);
and
one
flavanone,
eriodictyol
(4).
The
structures
of
1-‐4
were
elucidated
on
the
basis
of
1D
NMR
and
2D
NMR
correlations
and
comparison
with
previous
literature.
Acknowledgements:
Universiti
Teknologi
MARA
is
acknowledged
for
financial
and
technical
support
Keywords: Cynometra cauliflora L., chromatography, flavones, flavanone
References:
[1]
Seidemann
J.
World
spice
plants:
Economic,
Usage,
Botany,
Taxonomy.
Berlin-‐Heidelberg,
Springer
2005;
131
[2]
Ikram
EHK,
Eng
KH,
Jalil
AMM,
Ismail
A,
Idris
S,
Azlan
A,
Nazri
HSM,
Diton
NAM,
Mokhtar
RAM.
Antioxidant
capacity
and
total
phenolic
content
of
Malaysian
underutilized
fruit.
J
Food
Comp
Anal
2009;
22:
388-‐393
[3]
Aziz
AFA,
Iqbal
M.
Antioxidant
activity
and
phytochemical
composition
of
Cynometra
caul-‐
iflora.
J
Exp
Integr
Med
2013;
3:
337–341
[4]
Ado
MA,
Abas
F,
Mohammed
AS,
Ghazali
HM.
Anti-‐
and
pro-‐lipaseactivity
of
selected
me-‐
dicinal,
herbal
and
aquatic
plants,
andstructure
elucidation
of
an
anti-‐lipase
compound.
Molecules
2013;
18:
14651-‐14669
[5]
Sedgley
M,
Gardner
JA.
International
Survey
of
Underexploited
Tropical
and
Subtropical
Perennials.
Acta
Horticulturae,
Wageningen
1989:
206
[6]
Ado
MA,
Abas
F,
Ismail
IS,
Ghazali
HM,
Shaari
K.
Chemical
profile
and
antiacetylcholines-‐
terase,
antityrosinase,
antioxidant
and
α-‐glucosidase
inhibitory
activity
of
Cynometra
cauliflora
L.
Leaves.
J
Sci
Food
Agric;
Wiley
2014;
1-‐8
P399
Immunometabolic
regulation
by
triterpene-‐enriched
fraction
of
Eucalyptus
tereticornis
in
adipose
tissue
cell
line
models.
Sergio
Acin1,2,
Susana
Ceballos2,
Diana
L.
Muñoz1,
Adriana
Castaño3,
Fernando
Echeverri3,
Norman
Balcazar1,2.
1Department
of
Physiology
and
Biochemistry
-‐
School
of
Medicine,
2GENMOL
Group,
3QOPN
Group
-‐
School
of
Natural
and
Exacts
Sciences.
Sede
de
Investigación
Universitaria,
Universidad
de
Antioquia,
Calle
62
#
52-‐59,
Medellín,
Colombia
Obesity
is
associated
with
adipose
tissue
inflammation
that
eventually
results
in
insulin
re-‐
sistance
(IR)
and
it
is
linked
with
type
2
diabetes
mellitus
[1].
Previous
studies
established
the
beneficial
effects
of
fractions
of
Eucalyptus
tereticornis
(Eu)
on
carbohydrate
metabolism
in
a
diabetic
mouse
model.
In
addition,
they
showed
a
reduction
in
levels
of
proinflammatory
cy-‐
tokines
in
adipose
tissue
[2].
Our
aim
is
to
evaluate
the
effect
of
Eu
fractions
on
inflammation
and
IR
using
murine
adipocyte
and
macrophage
cell
lines.
A
crude
methanolic
extract
(OBE100)
from
leaves
of
Eu
was
partitioned
with
ethyl
acetate
and
purified
by
Sephadex
LH-‐20.
The
fractions
containing
triterpenes
(OBE104)
were
used
for
biological
assays.
3T3-‐L1
adipocytes
were
pre-‐treated
for
7
days
or
treated
for
24
h
after
dif-‐
ferentiation
with
OBE100
and
OBE104.
Fatty
acid
content
was
evaluated
by
oil
red
staining.
Macrophage
cell
line
J774,
activated
with
LPS
and
INFγ,
was
used
to
evaluate
OBE100
and
OBE104
anti-‐inflammatory
action.
Gene
expression
levels
of
TNFα,
IL6,
IL1β
and
MCP1
were
detected
by
real-‐time
RT-‐PCR.
In
addition,
protein
expression
levels
of
TNFα,
IL6
and
MCP1
were
monitored
by
flow
cytometry.
OBE100
and
OBE104
reduced
mRNA
and
protein
expression
of
pro-‐inflammatory
cytokines
MCP1,
IL1β,
IL6
and
TNFα
in
activated
macrophages.
Additionally,
a
reduction
of
the
intracel-‐
lular
fatty
acid
content
in
adipocytes
pre-‐treated
for
7
days
with
Eu
fractions
was
observed.
This
effect
was
not
detected
in
adipocytes
treated
for
24
h
after
differentiation.
These
results
suggest
that
Eu
fractions
inhibit
lipogenic
activity.
This
study
demonstrates
the
role
of
triterpenes
from
Eu
on
the
regulation
of
immunometabolic
response
in
cells
present
in
adipose
tissue.
Our
results
allow
us
to
suggest
that
the
systemic
effects
observed
on
carbohy-‐
drate
metabolism
with
the
treatment
of
fractions
of
Eu
could
be
explained
by
their
action
on
the
inflammation
process
established
in
adipose
tissue
of
obese
mice.
Acknowledgements: This research was supported by the Colciencias grant 111565740656
Keywords: Eucalytus Spp., Anti-‐inflammatory, Triterpenes, adipocytes, macrophages.
References:
[1] Lumeng
CN,
Deyoung
SM,
Bodzin
JL,
Saltiel
AR.
Increased
inflammatory
properties
of
adi-‐
pose
tissue
macrophages
recruited
during
diet-‐induced
obesity.
Diabetes
2007;
56:
16–23
[2] Guillén
A,
Granados
S,
Rivas
KE,
Estrada
O,
Echeverri
F,
Balcázar,
N.
Antihyperglycemic
ac-‐
tivity
of
Eucalyptus
tereticornis
in
insulin
resistant
cells
and
a
nutritional
model
of
diabet-‐
ic
mice.
Adv
Pharm
Sci
2015;
1-‐10
P400
Evaluation
of
the
hypoglycemic
effects
of
extracts
and
diterpe-‐
noids
from
Azorella
compacta
(llareta)
Aurelio
San-‐Martín1,
Mitchell
Bacho1,
Sylvian
Cretton2,
Philippe
Christen2,
Andres
Olea3,
Diana
Muñoz4,
Alis
Guillen4,
Norman
Balcazar4
1
Facultad
de
Ciencias.
Universidad
de
Chile.
Las
Palmeras
3425.
Santiago
de
Chile,
Chile;
2
University
of
Geneva,
Quai
Ernest-‐Ansermet
30.
Genève,
Switzerland;
3
Instituto
de
Ciencias
Químicas
aplicadas
Univer-‐
sidad
Autónoma
de
Chile.
Llano
Subercaseaux
2801
Santiago
Chile;
4
Department
of
Physiology
and
Bio-‐
chemistry
-‐
School
of
Medicine,
Sede
de
Investigación
Universitaria,
Universidad
de
Antioquia,
Calle
62
#
52-‐59,
Medellín,
Colombia
Aqueous
or
ethanol
infusions
of
llareta
have
been
used
as
antidiabetics
in
the
popular
medi-‐
cine
in
the
Chilean
highlands.
A
previous
report
[1]
demonstrated
the
hypoglycemic
effect
of
some
compounds
on
a
type
1
diabetic
rat
model.
In
order
to
determine
the
compounds
re-‐
sponsible
for
this
effect,
we
evaluated
the
hypoglycemic
activity
of
llareta
crude
extract
and
some
of
its
constituents,
on
the
C2C12
cell
line.
The
leaves
of
Azorella
compacta
were
collect-‐
ed
in
Tatio,
Antofagasta
region,
Chile.
The
plant
extract
was
prepared
using
petroleum
ether,
dichloromethane
and
methanol.
The
extract
was
fractionated
using
chromatographic
tech-‐
niques
with
Sephadex
LH-‐20
and
silica
gel.
Three
diterpenes
were
isolated
and
identified
by
NMR
analysis.
The
crude
extract
and
diterpenes
were
incorporated
into
micelles
using
an
aqueous
solution
of
the
polymer
Pluronic
F127
(1mM)
for
solubilisation.
C2C12
myotubes
were
incubated
with
different
concentrations
of
Azorella
crude
extract
and
the
3
diterpenes
(Y1,
Y2
and
Y5)
for
4h,
in
glucose
uptake
medium.
The
crude
extract
and
Y5
diterpene
stimu-‐
lated
glucose
uptake
in
C2C12
myotubes
up
to
50%.
The
effect
was
also
evaluated
using
a
cell
line
model
of
insulin
resistance.
C2C12
insulin-‐resistant
cells
were
established
by
adding
palmitic
acid
(0.75mM)
to
the
culture
medium
and
then
cells
were
treated
with
the
crude
ex-‐
tract
or
Y5.
Both
treatments
increased
glucose
uptake
in
C2C12
insulin-‐resistant
cells
and
displayed
an
effect
similar
to
metformin,
used
as
a
positive
control.
Glucose
consumption
of
C2C12
cells
was
determined
by
a
glucose
oxidase
method.
The
findings
of
this
study
are
in
agreement
with
the
traditional
use
of
the
leaves
of
llareta
as
a
hypoglycemic
agent.
We
have
shown
that
one
of
the
substances
responsible
for
the
effects
is
a
diterpene
that
significantly
stimulated
glucose
uptake
in
C2C12
under
conditions
of
insulin
resistance.
Acknowledgment:
This
research
was
supported
by
Fondecyt
project
Nº
1120199
and
Sistema
General
de
Regalias
Colombia
No.
2013000100177
Keywords: Azorella compacta, Anti-‐diabetic, diterpenes, C2C12 myotubes, insulin resistance.
References:
[1] Fuentes
NL,
Sagua
H,
Morales
G,
Borquez
J,
San
Martin
A,
Soto
J,
Loyola
LA.
Experimental
antihyperglycemic
effect
of
diterpenoids
of
Llareta
Azorella
compacta
(Umbelliferae)
Phil
in
rats.
Phytother
Res
2005;
19:
713–716
P401
New
sesquiterpene
lactones
from
Ambrosia
cumanensis
Kunth.
N.
Malafronte1,
R.
Cotugno1,
N.
Jimenez2,
M.
De
Leo3,
A.
Braca3,
N.
De
Tommasi1
1
Dipartimento
di
Farmacia,
Università
di
Salerno,
Via
Giovanni
Paolo
II,
84084
Fisciano,
Salerno,
Italy,
2
Grupo
de
Investigacion
en
Sustancias
Bioactivas,
Facultad
de
Ciencias
Farmaceuticas
y
Alimentarias,
Universidad
de
Antioquia
UdeA,
Calle
70
No.
52-‐21,
Medellin,
Colombia,
3
Dipartimento
di
Farmacia,
Uni-‐
versità
di
Pisa,
Pisa,
Via
Bonanno
33,
56126
Pisa,
Italy
Sesquiterpene
lactones
are
a
wide
and
various
group
of
natural
compounds
showing
a
large
spectrum
of
biological
activities,
including
antitumor,
anti-‐inflammatory,
antibacterial
and
antifungal
activities
[1].
These
compounds
are
mainly
distributed
in
the
Asteraceae
family.
The
genus
Ambrosia
(Asteraceae)
includes
41
species
widespread
in
the
temperate
regions
of
the
Northern
Hemisphere
and
South
America
but
many
of
which
are
adapted
to
the
arid
cli-‐
mates
of
the
desert.
The
genus
is
best
known
for
the
severe
and
widespread
allergies
caused
by
its
pollen.
A.
cumanensis
Kunth
is
a
medicinal
plant
native
of
Central
and
South
America.
The
leaves
are
used
in
Colombian
traditional
medicine
for
stomach
pain,
intestinal
infections,
gastritis,
and
muscle
aches.
Previous
biological
studies
on
extracts
obtained
from
A.
cumanen-‐
sis
leaves,
reported
antimicrobial
and
spasmolitic
activity
[2].
On
the
basis
of
our
previous
studies
on
plants
belonging
to
the
Ambrosia
genus
[3,
4]
and
some
biological
activities
report-‐
ed,
a
phytochemical
investigation
on
A.
cumanensis
aerial
parts
was
performed.
The
dried
powdered
leaves,
collected
in
Eastern
Antioquia
region
of
Colombia,
were
extracted
with
MeOH
and
partitioned
between
n-‐butanol/water
to
remove
sugars.
The
n-‐BuOH
extract
was
dried
and
dissolved
in
chloroform
and
subjected
to
different
chromatographic
techniques
such
as
Silica
gel,
MPLC
and
RP-‐HPLC.
Eleven
sesquiterpene
lactones,
including
three
new
natural
products
(1-‐3)
were
characterized
by
1D-‐
and
2D-‐NMR
and
MS
analyses.
New
com-‐
pounds
were
tested
for
their
antiproliferative
activity
on
HeLa
(cervical
cancer),
Jurkat
(T
lymphocyte),
and
U937
(lymphoma)
cell
lines.
Compound
3
(2,
3-‐dehydropsilostachyn
C),
was
found
to
be
the
most
active
showing
an
IC50
6
μM
in
Jurkat
and
8
μM
in
U937,
respectively.
OH
O O
HO HO
O O O
O O
1 O 2 3
Keywords: Ambrosia cumanensis, Asteraceae, sesquiterpene lactones, cytotoxic activity
References:
[1] Robles
M,
Aregullin
M,
West
J,
Rodriguez
E.
Recent
studies
on
the
zoopharmacognosy,
pharmacology
and
neurotoxicology
of
sesquiterpene
lactones.
Planta
Med
1995;
61:
199-‐
203
[2] Jimenez
N,
Carillo-‐Hormaza
L,
Pujol
A,
Alzate
F,
Osorio
E,
Lara-‐Guzman
O.
Antioxidant
ca-‐
pacity
and
phenolic
content
of
commonly
used
anti-‐inflammatory
medicinal
plants
in
Co-‐
lombia.
Ind
Crop
Prod
2015;
70:
272-‐279
[3] De
Leo
M,
Vera
Saltos
MB,
Naranjo
Puente
BF,
De
Tommasi
N,
Braca
A.
Sesquiterpenes
and
diterpenes
from
Ambrosia
arborescens.
Phytochemistry
2010;
71:
804-‐809
[4] Cotugno
R,
Fortunato
R,
Santoro
A,
Gallotta
D,
Braca
A,
De
Tommasi
N,
Belisario
MA.
Effect
of
sesquiterpene
lactone
coronopilin
on
leukemia
cell
population
growth,
cell
type-‐
specific
induction
of
apoptosis
and
mitotic
catastrophe.
Cell
Proliferat
2012;
45:
53-‐65
P402
Antioxidant
capacity
of
Crataegus
monogyna
and
Crataegus
pen-‐
tagyna
leaves
and
fruits
harvested
from
the
Danube
Delta
Oana
C.
Bujor1,2,3,
Camelia
P.
Stefanache1,
Irina
Volf3,
Doina
Danila1
1NIRDBS
/
“Stejarul”
Biological
Research
Centre,
610004
Piatra
Neamt,
Romania,
2UMR408
SQPOV,
INRA,
University
of
Avignon,
F-‐84000
Avignon,
France,
3Faculty
of
Chemical
Engineering
and
Environmental
Protection,
“Gheorghe
Asachi”
Technical
University
of
Iasi,
700050
Iasi,
Romania
Crataegus
species
are
known
to
be
used
in
natural
health
products
like
tinctures,
tablets
and
teas
for
the
treatment
of
heart
diseases
[1].
In
vitro,
the
fruits,
leaves
and
flowers
of
Crataegus
species
have
also
been
shown
to
exhibit
antioxidant
and
antiproliferative
activities
[2,
3].
How-‐
ever,
these
properties
are
influenced
by
the
type
of
plant
organ,
the
harvest
season,
the
spe-‐
cies
and
the
geographical
origin.
In
this
contex,
different
extracts
of
leaves
and
ripe
fruits
of
Crataegus
monogyna
(CM)
and
C.
pentagyna
(CP)
collected
from
natural
habitats
on
the
limit
of
Danube
Delta
Biosphere
Reserve
in
September
2015
were
evaluated
for
their
antioxidant
capacity.
The
aqueous,
40%
(EtOH40)
and
70%
(EtOH70)
ethanolic
extracts
were
obtained
from
the
dried
plant
material
by
ultrasound-‐assisted
extraction.
The
antioxidant
content
of
extracts
was
determined
by
Folin-‐Ciocalteu
method,
while
the
radical
scavenging
activity
was
assessed
by
DPPH
test.
In
ethanolic
extracts,
the
content
of
antioxidants
ranges
between
11.2
(EtOH40,
fruits)
and
28.7
(EtOH40
and
EtOH70,
leaves)
mg
GAE/g
(mg/g
gallic
acid
equivalents)
for
CM
extracts,
while
for
CP
extracts
it
varied
between
8.95
(EtOH40,
fruits)
and
32.6
(EtOH70,
leaves)
mg
GAE/g.
Similar
DPPH
activities
(2.34
µM
TE/g,
micromoles
of
Trolox
Equivalents)
were
found
for
leaf
EtOH
extracts
of
CM
and
CP.
The
EtOH
extracts
of
CM
fruits
presented
stronger
DPPH
activity
(2.07
µM
TE/g)
compared
to
the
EtOH
extracts
of
CP
fruits
(1.46
µM
TE/g
for
EtOH40
and
1.49
µM
TE/g
for
EtOH70).
The
aqueous
extracts
of
both
Crataegus
spe-‐
cies
showed
a
lower
antioxidant
capacity.
In
the
tested
extracts,
chlorogenic
acid,
hyperoside,
isoquercitrin,
and
rutin
were
detected
by
TLC-‐fingerprinting.
Isoquercitrin
and
rutin
were
not
detected
in
CP
leaves
and
fruits
and
CM
leaves,
respectively.
In
conclusion,
the
results
of
this
study
indicate
that
leaves
and
fruits
of
Crataegus
species
provide
considerable
antioxi-‐
dant
protection
and
can
be
suitable
raw
material
for
the
food
and
cosmetic
industries.
Acknowledgements:
The
work
was
sustained
from
the
project
PN
09-‐360401
(BIODIV)
financed
by
MEN-‐
ANCS,
Romania
Keywords:
Folin-‐Ciocalteu,
DPPH,
ethanolic
extract,
aqueous
extract,
phenolic
compounds
References:
[1] Edwards
JE,
Brown
PN,
Talent
N,
Dickinson
TA,
Paul
R.
Shipley
PR.
A
review
of
the
chemis-‐
try
of
the
genus
Crataegus.
Phytochemistry
2012;
79:
5–26
[2] Giurescu
Bedreag
CF,
Trifan
A,
Bucur
LA,
Arcus
M,
Tebrencu
C,
Miron
A,
Costache
II.
Chemi-‐
cal
and
antioxidant
studies
on
Crataegus
pentagyna
leaves
and
flowers.
Rom
Biotech
Lett
2014;
19:
9859−9867
[3] Rodrigues
S,
Calhelha
RC,
Barreira
JCM,
Dueñas
M,
Carvalho
AM,
Abreu
RMV,
Santos-‐Buelga
C,
Ferreira
ICFR.
Crataegus
monogyna
buds
and
fruits
phenolic
extracts:
Growth
inhibitory
activity
on
human
tumor
cell
lines
and
chemical
characterization
by
HPLC–DAD–ESI/MS.
Food
Res
Int
2012;
49:
516–523
P403
Sesquiterpene
lactones
and
phenolic
compounds
content
in
Arni-‐
ca
montana
flowers
and
leaves
samples
harvested
from
wild
sites
in
North-‐East
Romania
Camelia
P.
Stefanache1,
Oana
C.
Bujor1,2,3,
Radu
Necula1,4,
Valentin
Grigoras1,
Constantin
Mardari5,
Ciprian
Birsan5,
Doina
Danila1
1
NIRDBS
/
“Stejarul”
Biological
Research
Centre,
Alexandru
cel
Bun
no.
6,
610004
Piatra
Neamt,
Roma-‐
nia,
2
Faculty
of
Chemical
Engineering
and
Environmental
Protection,
“Gheorghe
Asachi”
Technical
Uni-‐
versity
of
Iasi,
700050
Iasi,
Romania,3
UMR408
SQPOV,
INRA,
Avignon
University,
F-‐84000
Avignon,
France,4
Faculty
of
Chemistry,”Alexandru
Ioan
Cuza”
University
of
Iasi,
700506
Iasi,
Romania,5
“A.
Fatu”
Botanica
Garden,
”Aexandru
Ioan
Cuza”
University
of
Iasi,
700506
Iasi,
Romania.
Our
study
aimed
to
assess
the
phenolic
compounds
and
sesquiterpene
lactones
(SL)
content
in
the
flowers
and
leaves
of
Arnica
montana
collected
from
wild
populations
at
different
alti-‐
tudes:
5
sites
at
800
-‐1000
m
and
5
sites
at
1000
-‐
1700
m.
The
dried
plant
materials
were
extracted
with
a
mixture
of
methanol/acetone/water
for
the
extraction
of
the
phenolic
com-‐
pounds.
The
method
from
the
European
Pharmacopoeia
[1]
was
used
for
the
determination
of
the
SLs.
The
separation
and
quantification
of
the
phenolic
compounds
and
SLs
was
performed
by
HPLC-‐DAD
analysis.
In
flowers,
the
following
phenolic
compounds
were
identified:
chlorogenic
acid,
caffeic
acid,
cynarin
and
the
flavonoids
isoquercitrin,
apigenin
and
apigenin-‐7-‐O-‐glucoside.
In
the
leaf
samples,
we
found
chlorogenic
acid
and
cynarin,
while
from
the
group
of
flavonoids
only
apigenin
was
identified.
In
both
flower
and
leaf
samples,
cynarin
was
the
major
phenolic
acid
(4.27–9.51
mg/g
d.w.
in
flowers
and
6.44–8.65
mg/g
in
leaves),
while
isoquercitrin
was
the
major
flavonoid
in
flowers
(1.66–3.90
mg/g).
The
leaves
had
fewer
flavonoids
and
in
signifi-‐
cantly
lower
amounts.
The
SLs
content
in
flowers
varied
from
1.02
to
1.59
%,
and
from
0.31–
0.51%
in
the
leaves.
Flowers
collected
at
altitudes
of
1000
-‐
1700
m
had
a
higher
content
of
phenolic
acids
(17.51-‐
21.25
mg/g),
flavonoids
(7.58–12.17mg/g)
and
SLs
(1.36–1.59%),
compared
to
the
samples
from
lower
altitudes
with
11.05–21.51
mg/g
phenolic
acids,
7.58–12.17
mg/g
flavonoids
and
1.02–1.40%
SLs,
respectively.
In
contrast,
the
amounts
for
the
bioactive
compounds
groups
were
lower
in
the
leaf
samples
harvested
at
higher
altitudes.
The
content
of
phenolic
com-‐
pounds
and
SLs
is
similar
with
literature
data
[2-‐4].
The
variability
of
the
bioactive
com-‐
pounds
content
may
be
due
to
the
reaction
of
the
plants
to
the
environmental
peculiarities
of
the
sites
generated
by
the
altitude
gradient
(vegetation,
phytophagous
species,
meteorological
and
climatic
factors,
soil,
and
slope).
Acknowledgements:
The
work
was
conducted
within
the
Program
Partnership
in
Priority
Area
-‐
PNII
supported
byMEN-‐UEFISCDI,
Project
No.
74/2014.
Keywords:
Altitude
gradient,
Arnica
montana,
phenolic
acids,
flavonoids,
sesquiterpene
lac-‐
tones.
References:
[1] European
Pharmacopoeia
7.0
[2] Stefanache
CP,
Peter
S,
Meier
B,
Danila
D,
Tanase
C,
Wolfram
E.
Phytochemical
composition
of
Arnicae
flos
from
wild
populations
in
the
northern
area
of
the
Romanian
Eastern
Carpa-‐
thians.
Rev
Chim
2015;
66:
784
–
787
[3] Aiello
N,
Bontempo
R,
Vender
C,
Ferretti
V,
Innocenti
G,
Dall`Acqua
S.
Morpho-‐quantitative
and
qualitative
traits
of
Arnica
montana
L.
wild
accessions
of
Trentino,
Italy.
Industrial
Crops
and
Products
2012;
40:
127-‐140
[1] Ganzera
M,
Egger
C,
Zidorn
C,
Stuppner
H.
Quantitative
analysis
of
flavonoids
and
phenolic
acids
in
Arnica
montana
L.
by
micellar
electrokinetic
capillary
chromatography.
Anal
Chim
Acta
2008;
614:
196-‐200
P404
Investigation
of
potential
anti-‐diabetic
effect
of
Mucuna
pruriens
(L)
DC
(Fabaceae)
aqueous
leaf
extract
Oke-‐Oghene
P.
Akpoveso1,
Vesna
Tumbas-‐Saponjac2,
Prabal
K.
Chatterjee1,
George
Olivier1
1 School of Pharmacy and Biomolecular Sciences, University of Brighton, Brighton, BN2 4GB, United King-‐
dom; 2 Faculty of Technology, University of Novi Sad, Bulevar cara Lazara 1, 21000 Novi Sad, Serbia.
Mucuna
pruriens
(L.)
DC
is
an
herbal
plant
popularly
used
in
parts
of
Africa
(incl
Nigeria,
where
it
is
taken
as
a
tea)
for
the
treatment
of
anaemia
and
in
some
parts
of
Asia
for
treat-‐
ment
of
diabetes
[1,2].
The
antidiabetic
effect
of
alcoholic
leaf
extract
of
Mucuna
pruriens
(MP)
has
been
studied
previously
in
diabetic
rats
[3];
however,
the
potential
antidiabetic
effect
of
an
aqueous
extract
has
not
been
evaluated.
For
this
study,
MP
leaves
were
collected
and
iden-‐
tified
and
stored
at
the
herbarium
in
the
International
Centre
for
Ethnomedicine
and
Drug
development,
Enugu
State,
Nigeria,
West
Africa.
The
specimen
identification
number
is:
Inter-‐
CEDD-‐16018.
We
investigated
the
effect
of
an
aqueous
extract
of
Mucuna
pruriens
leaves
(MPLE)
on
glucose
uptake
in
rat
NRK-‐52E
renal
cell
line.
Glucose
uptake
was
measured
using
fluorescent
glucose
(2-‐NBDG).
Phenolics
in
MPLE
were
detected
using
HPLC-‐UV/VIS.
The
peaks
where
identified
by
comparison
with
reference
peaks
obtained
for
known
flavonoids
and
phenolic
acids.
The
results
as
shown
in
the
figures
below
indicate
that
MPLE
contains
phenolics,
and
it
can
also
limit
glucose
re-‐uptake.
Identified
phenolics
included
Epicatechin,
Rutin,
Gallic
acid,
Caffeic
acid,
Coumaric
acid,
and
Ellagic
acid.
1mg/ml
MPLE
inhibited
glu-‐
cose
uptake
in
NRK-‐52E
cell
lines.
This
effect
was
comparable
to
1mM
Phloridzin
(a
standard
sodium
glucose
transporter
inhibitor).
Glucose
uptake
inhibition
is
useful
for
insulin
inde-‐
pendent
control
of
hyperglycaemia
via
the
kidneys
as
well
as
in
the
intestines
[4].
Effect of Mucuna pruriens leaf extract and Phlorizin on glucose uptake in NRK-52E cell line
5000
Flourescence (glucose uptake)
*
4000
3000
2000
1000
Identification of phenolics: 1: Gallic acid, 2: Vanillic
0
ly
k
M
an
PL
m
on
1m
0m
25
Bl
lM
acid, 3: Catechin, 4: Protocatechuic acid, 5: Caffeic acid,
0.
1.
DG
0.
m
DZ
DZ
NB
g/
PH
PL
PL
1m
Treatment
6:
Epicatechin,
7:
Ellagic
acid.
References:
[1] Lampariello
LR,
Cortelazzo
A,
Guerranti
R,
Sticozzi
C,
Valachi
G.
The
magic
velvet
bean
of
Mucuna
pruriens.
J.
Tradit
Complement
Med
2012;
2:
331-‐339
[2] Akindele
AJ,
Busayo
FL.
Effects
of
the
hydroethanolic
extract
of
Mucuna
pruriens
(L.)
DC
(Fabaceae)
on
haematological
profile
in
normal
and
haloperidol
treated
rats.
Nig
Q
Hosp
Med
2011;
21:
93-‐98
[3] Murugan
M,
Reddy
CUM.
Hypoglycemic
and
hypolipidemic
activity
of
leaves
of
Mucuna
pruriens
in
alloxan
induced
diabetic
rats.
J
Pharm
Sci
Technol
2009;
1:
69-‐73
[4] Mudaliar
S,
Polidori
D,
Zambrowicz
B,
Henry
RR.
Sodium-‐glucose
cotransporter
inhibitors:
Effects
on
renal
and
intestinal
glucose
transport.
Diabetes
care
2015;
38:
2344-‐2345
P405
Phytochemical
study
of
Helleborusodorus
subsp.
cyclophyllus
(Ranunculaceae)
Olga
St.
Tsiftsoglou1,
Eirini
N.
Kalpourtzi1,
Michalis
K.
Stefanakis1,2,
Diamanto
M.
Lazari1
1Aristotle
University
of
Thessaloniki,
School
of
Pharmacy,
Laboratory
of
Pharmacognosy,
54124
Thessa-‐
loniki,
Greece,
2University
of
Crete,
Department
of
Chemistry,
Laboratory
of
Organic
Chemistry,
71003
Heraklion,
Greece
Interest
in
the
genus
Helleborus
(Ranunculaceae),
has
recently
increased
because
it
is
a
known
source
of
metabolites
which
are
considered
as
promising
remedies
for
severe
diseases
such
as
cancer
and
diabetes
[1].
The
genus
comprises
around
20
species.
In
the
current
study,
extracts
of
the
leaves
and
roots
of
H.
odorussubp.
cyclophyllus
were
examined
for
their
chemi-‐
cal
substitutes.
So
far,
ten
secondary
metabolites
were
isolated
by
chromatographic
methods
(
V.L.C,
C.C,
HPLC
)
and
identified
by
using
spectroscopic
methods
(MS
and
NMR):two
phenolic
acids:
2-‐O-‐feruloyl-‐L-‐malate
(1)
and
8-‐O-‐Glucose-‐hydroxy
tyrosol
(2),
two
ecdysteroids:
5β-‐hydroxy-‐ecdysterone
(Polypodine
B)
(3)
and
20-‐hydroxy-‐ecdyson
(4),
two
bufadi-‐
enolides:5β,14β-‐dihydroxy-‐19-‐oxo-‐3β-‐[(α-‐L-‐rhamnopyranosyl)oxy]bufa-‐20,22-‐dienolide
(deglucohellebrin)(5)
and
(3β,5β)-‐3-‐[(6-‐Deoxy-‐4-‐O-‐β-‐D-‐glucopyranosyl-‐α-‐L-‐
mannopyranosyl)oxy]-‐5,14-‐dihydroxy-‐19-‐oxo-‐bufa-‐20,22-‐dienolide(hellebrin)
(6),
two
furo-‐
stanol
saponins:
furosta-‐5,20(22),25(27)-‐triene-‐1β,3β,11α,26-‐tetrol
26-‐O-‐β-‐D-‐
glucopyranoside
(Caucasicoside
A)
(7)
and
Helleboroside
B
(8),
as
well
as
Uridine
(9)
and
2-‐
deoxy-‐D-‐ribono-‐1,4-‐lactone(10).
It
is
the
first
time
that
compounds
(1),
(2),
(9)
and
(10)
are
reported
in
the
genus
Helleborus.
Acknowledgements:
Haralambos
Katerinopoulos
is
acknowledged
for
his
contribution
in
this
research
Keywords: Ranunculaceae, furostanols, phenolic acid, ecdysteroid, bufadienolide, HPLC
References:
[1] Maior
MC
and
Dobrotă
C.
Natural
compounds
with
important
medical
potential
found
in
Helleborus
sp.
Cent
Eur
J
Biol
2013;
8:
272-‐285
P406
Study
of
Mezoneuron
benthamianum,
a
plant
traditionally
used
against
malaria
in
Guinea
Alembert
T.
Tchinda1,2,†,
Jean
Loua3,4,†,
Virginie
Esters1,
Ewa
Cieckiewicz1,
Allison
Ledoux1,
Luc
Angenot1,
Monique
Tits1,
Aliou
M.
Balde3,4,
Michel
Frédérich1,
Olivia
Jansen1
1Laboratory
of
Pharmacognosy,
Department
of
Pharmacy,
CIRM,
University
of
Liège,
CHU
B36,
4000
Li-‐
ège,
Belgium;
2
Center
for
Studies
on
Medicinal
Plants
and
Traditional
Medicine,
Institute
of
Medical
Re-‐
search
and
Medicinal
Plants
Studies
(IMPM),
P.O.
Box
6163,
Yaoundé,
Cameroon;
3
Research
and
Valori-‐
zation
Center
on
Medicinal
Plants
of
Dubreka,
Dubreka,
Guinea;
4
Departement
of
Pharmacy,
University
Gamal
Abdel
Nasser
of
Conakry,
Conakry,
Guinea.
†
These
authors
contributed
equally
to
this
work
Despite
some
improvements
in
malaria
control,
this
parasitic
disease
remains
a
major
public
health
problem
in
many
African
countries,
causing
about
400
000
deaths/year
through
the
continent,
mainly
by
children
under
the
age
of
five
[1].
In
Guinea,
the
leaves
of
Mezoneuron
benthamianum
Baill.
are
traditionally
used
to
treat
malar-‐
ia
[2]
and
showed
a
good
antiplasmodial
activity
in
an
antiprotozoal
in
vitro
screening
[3],
as
well
as
promising
results
in
a
preliminary
small-‐scale
ethnomedical
study
(unpublished
data),
encouraging
us
to
continue
the
study
of
this
plant.
The
aim
of
this
work
was
to
evaluate
the
activity
of
M.
benthamianum
leaves
extracts
against
P.
falciparum
using
an
in
vitro
test
model
(p-‐LDH
assay)
and
to
undertake
a
bio-‐guided
fractionation
to
identify
the
compounds
respon-‐
sible
for
the
activity.
Hydroethanolic
extracts
(70%
v/v)
of
M.
benthamianum
leaves
showed
a
moderate
in
vitro
activity
against
P.
falciparum
3D7,
with
IC50
=
22.5
–
32.6
µg/ml,
depending
on
the
batch;
while
a
dark
precipitate
formed
during
ethanol
evaporation
showed
higher
ac-‐
tivity
(IC50
=
6,5µg/ml).
The
bioguided
fractionation
was
performed
on
this
most
active
frac-‐
tion
and
allowed
the
isolation
of
three
diterpens,
two
flavonoids,
resveratrol,
gallic
acid
and
its
ethylester,
β-‐sitosterol
glucoside
and
pheophorbide
derivatives.
The
active
compounds
belong
to
several
phytochemical
classes,
including
flavonoids,
pheophorbide
and
gallic
acid
derivatives,
contributing
together
to
the
global
antiplasmodial
activity
of
the
hydroalcoholic
extract
against
P.
falciparum
parasite.
This
study
gives
some
concrete
evidence
to
support
the
ethnopharmacological
use
of
Mezoneuron
benthamianum
leaves
extract
in
the
management
of
malaria.
The
active
compounds
can
be
further
studied
for
their
antiplasmodial
potential,
as
well
as
their
suitability
to
be
used
as
quality
markers
for
the
standardization
of
this
herbal
drug
from
the
Guinean
traditional
pharmacopeia.
Keywords:
Mezoneuron
benthamianum,
Antiplasmodial
activity,
Guinea
References:
[1] World
Health
Organization
(WHO),
World
Malaria
Report,
2015
[2] Traore
MS,
Baldé
MA,
Diallo,
MST,
Baldé
ES,
Diané
S,
Camara
A,
Diallo
A,
Balde
A,
Keïta
A,
Keita
SM,
Oularé
K,
Magassouba
FB,
Diakité
I,
Diallo
A,
Pieters
L,
Baldé
AM.
Ethnobotanical
survey
on
medicinal
plants
used
by
Guinean
traditional
healers
in
the
treatment
of
malar-‐
ia.
J
Ethnopharmacol
2013;
150:
1145–1153
[3] Traore
MS,
Diane
S,
Diallo
MST,
Balde
ES,
Balde
MA,
Camara
A,Diallo
A,
Keita
A,
Cos
P,
Maes
L,
Pieters
L,
Balde
AM.
In
Vitro
Antiprotozoal
and
Cytotoxic
Activity
of
Ethnopharmacolog-‐
ically
Selected
Guinean
Plants.
Planta
Med
2014;
80:
1340–1344
P407
In-‐vitro
bioassays
for
plant
extracts
with
tick-‐repellent
and
acari-‐
cidal
properties:
a
systematic
review
and
meta-‐analysis
Olubukola
Adenubi1,
Folorunso
Fasina2,
Lyndy
McGaw1,
Jacobus
Eloff1,
Vinny
Naidoo3
1Phytomedicine Programme, Department of Paraclinical Sciences, Faculty of Veterinary Sciences, Univer-‐
sity
of
Pretoria,
Onderstepoort
0110,
Pretoria,
South
Africa,
2Department
of
Veterinary
Tropical
Diseases,
Faculty
of
Veterinary
Sciences,
University
of
Pretoria,
Onderstepoort
0110,
Pretoria,
South
Afri-‐
ca,3Biomedical
Research
Centre,
Faculty
of
Veterinary
Sciences,
University
of
Pretoria,
Onderstepoort
0110,
Pretoria,
South
Africa.
Ticks
are
arthropods
which
transmit
pathogens
to
humans
and
animals
causing
great
eco-‐
nomic
losses
[1].
Chemical-‐based
anti-‐tick
measures
include
pyrethroids,
organophosphates,
amitraz
and
avermectins
with
significant
costs,
resistance
development
and
environmental
impacts
[2].
Plant-‐based
alternatives
may
have
high
efficacy,
low
toxicity
and
reduced
envi-‐
ronmental
impacts
[3].
We
collated
peer-‐reviewed
articles
on
plants
with
tick-‐repellent
and/or
acaricidal
properties
through
literature
searches
in
veterinary
databases
and
con-‐
ducted
meta-‐analyses
on
the
outcomes.
Bioassays
used
include:
tick
climbing
repellency,
olfactometry,
larval-‐packet
and
immersion
tests
[4].
Significant
differences
exist
in
the
methods
and
their
outcomes.
Meta-‐analysis
was
conducted
using
the
Fixed-‐effect
model
in
an
Excel
programme.
Using
a
total
of
1,428;
1,924;
574;
281
and
68
events,
the
median
efficiency
value
(MEV)
for
acaricidal,
larvicidal,
egg
hatch-‐
ing
inhibition,
inhibition
of
oviposition,
repellency,
acaricidal
effects
of
the
Lamiaceae
and
Asteraceae
families
were
80.12%
(CI95%:
79.20
–
81.04),
86.05%
(CI95%:
85.13
–
86.97),
83.39%
(CI95%:
82.47
–
84.31),
53.01%
(CI95%:
52.08
–
53.93),
92.00%
(CI95%:
91.08
–
92.93),
80.79%
(CI95%:
79.87
–
81.71)
and
48.34%
(CI95%:
47.42
–
49.26)
respectively.
The
plants
displayed
very
good
to
excellent
acaricidal,
larvicidal,
egg
hatching
inhibition,
and
repellency
inhibition
effects
but
relatively
poor
inhibition
of
oviposition.
However,
there
was
minimum
to
wide
disparities
compared
to
the
MEV
for
each
category.
Approximately
63%
and
69%
of
all
the
plants
evaluated
for
acaricidal
and
larvicidal
assays
out-‐surpass
the
MEV
for
all
plants
assayed
respectively.
While
ethnoveterinary
plants
hold
huge
potential
as
para-‐
siticides,
standardization
of
components,
extraction
techniques
and
experimental
design
is
mandatory,
as
well
as
mammalian
toxicological
profiling
and
excipient
development.
Similar-‐
ly,
investigation
of
the
residual
activities
and
shelf-‐life
of
these
plants
are
required
to
maxim-‐
ise
their
potential.
Translational
research
should
also
be
used
to
convert
in
vitro
studies
to
in
vivo
tests
and
the
possibility
of
drug
resistance
associated
with
these
promising
alternatives
should
be
explored.
Acknowledgements:
Financial
support
from
Technology
and
Innovation
Agency
(TIA)
in
collaboration
with
Council
for
Scientific
and
Industrial
Research
(CSIR,
Pretoria),
University
of
Pretoria,
L’Oreal-‐
UNESCO
Fellowship
and
the
Schlumberger
Faculty
for
the
Future
Foundation
is
thankfully
acknowledged.
Keywords:
acaricidal,
larvicidal,
egg
hatching
inhibition,
repellency,
bioassays,
medicinal
plants
References:
[1] Zahir
AA,
Rahuman
AA,
Bagavan
A,
Santhoshkumar
T,
Mohamed
RR,
Kamaraj
C,
Rajakumar
G,
Elango
G,
Jayaseelan
C,
Marimuthu
S.
Evaluation
of
botanical
extracts
against
Haema-‐
physalis
bispinosa
Neumann
and
Hippobosca
maculata
Leach.
Parasitol
Res
2010;
107:
585-‐592
[2] Patarroyo
JH,
Vargas
MI,
Gonzáles
CZ,
Gusmán
F,
Martins-‐Filho
OA,
Afonso
LCC,
Valente
FL,
Peconick
AP,
Marciano
AP,
Patarroyo
VAM,
Sossai
S.
Immune
response
of
bovines
stimu-‐
lated
by
synthetic
vaccine
SBm7462®
against
Rhipicephalus(Boophilus)
microplus.
Vet
Parasitol
2009;
166:
333–339
[3] Kayaa
GP.
The
potential
for
antitick
plants
as
components
of
an
integrated
tick
control
strategy. Ann
N
Y
Acad
Sci
2000; 916:
576-‐582
[4] Drummond
RO,
Crust
SF,
Trevino
JL,
Gladney
WJ,
Graham
OH.
B.
annulatus
and
B.
de-‐
colaratus:
laboratory
tests
of
insecticides.
J
Econ
Entomol
1973;
66:
130–133
P408
Toxicological
and
antimicrobial
studies
of
ethanolic
leaf
extract
of
Ricinodendron
heudelotii
(Baill.)
Heckel
Omolara
F.
Yakubu1,
Abiodun
H.
Adebayo1,
Olubanke
O.
Ogunlana1,
Jacob
O.
Popoola1,
Temi-‐
tope
A.
Ishola1,
Loretta
O.
Imonikhe1,
Oladipupo
A.
Adeyemi1
1Medicinal
Plant
Research
Group,
Department
of
Biological
Sciences,
College
of
Science
and
Technology,
Covenant University, P.M.B. 1023, Canaanland Ota, Ogun State, Nigeria
This research work was designed to evaluate the toxicological and antimicrobial activity of
ethanolic leaf extract of Ricinodendron heudelotii (Baill) Heckel. The antimicrobial activity of
the extract was investigated against 9 different bacteria species; Klebsiella sp, Shigella sp,
Escherichia coli, Salmonella sp(Gram-‐negatives), Staphylococcus sp, Bacillus sp, Micrococcus sp,
Streptococcus sp(Gram-‐positive) and Candida sp. (fungus) by agar well diffusion method and
minimum inhibitory concentration (MIC) was determined.1 Sub-‐acute toxicological effects of
the extract were evaluated on albino rats for 28 days. Five groups of seven male rats per
group were used for the study. Extract doses from 250 to 2000 mg/kg body weight were oral-‐
ly administered daily for 28 days.2 Animals were subsequently anaesthetized, blood samples
were collected for biochemical and haematological assays; liver was isolated, weighed and
processed for histological studies.3 A preliminary phytochemical screening showed that the
plant extract contains tannins, polyphenols, terpenoids, glycosides and alkaloids. Antimicro-‐
bial activity was observed against 7 of the 9 microorganisms tested with zone of inhibition
and MIC ranging from 18 to 36mm and 31.25 to 62.5mg/mL, respectively. Alanine transami-‐
nase, aspartate aminotransferase, alkaline phosphatase, urea, cholesterol, HDL cholesterol
and total bilirubin were significantly (p < 0.05) increased in higher doses of the extract while
there was no significant change in total protein and creatinine levels. A dose-‐ dependent re-‐
duction was observed in platelet count, haemoglobin, haematocrit and red blood cell count.
The toxicity exerted by ethanolic leaf extract of R. Heudelotii was confirmed from histological
sectioning which indicated sinusoidal congestion and severe vacuolar degeneration of
hepatocytes. The result suggests that the extract could serve as an effective antibiotic but the
prolong use of the extract may induce liver cell injury.
Acknowledgements: Biological Science Department is acknowledged for providing laboratory space
Keywords: Ricinodendron heudelotii, biochemical, haematological and histopathological pa-‐
rameters
References:
[1] Benkeblia
N.
Antimicrobial
activity
of
essential
oil
extracts
of
various
onions
(Allium
cepa)
and
garlic
(Allium
sativum).
LebensmWissTechnol.
2004;
37:
263-‐268
[2] Adebayo
AH,
Yakubu
OF,
Balogun
TM.
Protective
properties
of
Citrullus
lanatus
on
carbon
tetrachloride
induced
liver
damage
in
rats.
Eur
J.
Med
Plants
2014;
4:
979-‐989
[3] Aliyu
R,
Adebayo
AH,
Gatsing
D
and
Garba
IH.
The
effects
of
ethanolic
leaf
extract
of
Com-‐
miphora
Africana
(Burseraceae)
on
rat
liver
and
kidney
functions.
J.
Pharm
Toxicol
2007;
2:
373-‐379
P409
Lupinifolin
extracted
from
Derris
reticulata
inhibits
growth
of
Staphylococcus
aureus
possibly
by
damaging
bacterial
cell
mem-‐
brane
Kamol
Yusook1,
Oratai
Weeranantanapan2,
Siriporn
Riyajan1,
Jidapa
Musika1,
Nuannoi
Chud-‐
apongse1
1
School
of
Pharmacology,
Institute
of
Science,
Suranaree
University
of
Technology,
Nakhon
Ratchasima
30000,
Thailand,
2
School
of
Anatomy,
Institute
of
Science,
Suranaree
University
of
Technology,
Nakhon
Ratchasima
30000,
Thailand
Lupinifolin
is
a
prenylated
flavanone
that
has
been
isolated
from
several
medicinal
plants,
such
as
Myriopteron
extensum
[1],
Eriosema
chinense
[2]
and
Albizia
myriophylla
[3].
It
is
also
reported
to
be
a
major
compound
of
Derris
reticulata
[4].
In
the
present
study,
lupinifolin
was
isolated
from
Derris
reticulata
stem
and
identified
by
NMR
spectra.
Because
of
its
nonpolar
structure,
lupinifolin
is
very
soluble
in
organic
solvents,
but
sparingly
soluble
in
water.
Esti-‐
mated
from
log
Kow
(Octanol-‐Water
Partition
Coefficient),
water
solubility
of
lupinifolin
at
25
°C
is
0.009
mg/L
[5].
When
dissolved
in
alcohol
or
dimethyl
sulfoxide,
it
precipitates
after
di-‐
luted
in
aqueous
buffer.
In
this
study,
to
avoid
the
precipitation
in
aqueous
media,
lupinifolin
was
freshly
prepared
by
solubilizing
in
0.5
N
NaOH
and
immediately
diluted
in
Müller-‐Hinton
broth
(MHB)
for
antibacterial
test.
From
two-‐fold
microdilution
method,
it
was
found
that
lupinifolin
possessed
antimicrobial
activity
against
Staphylococcus
aureus
with
MIC
and
MBC
of
8
and
16
µg/mL,
respectively.
The
antibacterial
activity
of
lupinifolin
was
confirmed
by
TEM.
After
incubation
with
MIC
overnight,
lupinifolin
significantly
decreased
the
number
of
bacteria
compared
to
control
and
ruptured
bacterial
cell
membrane
and
cell
wall.
We
hypoth-‐
esize
that
lupinifolin
may
alter
bacterial
membrane
structure
and
function
similar
to
plant
flavonoids
previously
reported
[6].
Confirmation
of
the
mechanism
underlying
this
activity
of
lupinifolin
is
in
progress.
Acknowledgements:
This
investigation
was
supported
by
SUT
Research
and
Development
Fund.
We
thank
Dr.
Paul
J.
Grote
for
verification
of
plant
botanical
classification.
Keywords: Derris reticulata, lupinifolin, Staphylococcus aureus, antimicrobial, cell membrane
References:
[1] Soonthornchareonnon
N,
Ubonopas
L,
Kaewsuwan
S,
Wuttiudomlert
M.
Lupinifolin,
a
bioac-‐
tive
flavanone
from
Myriopteron
extensum
(Wight)
K.
Schum.
stem.
Thai
J.
Phytopharm
2004;
11:
19-‐27
[2] Prasad
SK,
Laloo
D,
Kumar
M,
Hemalatha
S.
Antidiarrhoeal
evaluation
of
root
extract,
its
bioactive
fraction,
and
lupinifolin
isolated
from
Eriosema
chinense.
Planta
Med
2013;
79:
1620-‐1627
[3] Joycharat
N,
Thammavong
S,
Limsuwan
S,
Homlaead
S,
Voravuthikunchai
SP,
Yingyong-‐
narongkul
BE,
Dej-‐Adisai
S,
Subhadhirasakul
S.
Antibacterial
substances
from
Albizia
myriophylla
wood
against
cariogenic
Streptococcus
mutans.
Arch
Pharm
Res
2013;
36:
723-‐730
[4] Chivapat
S,
Chavalittumrong
P,
Attiwist
A,
Soonthornchareonnon
N.
Toxicity
study
of
lupin-‐
ifolin
from
stem
of
Derris
reticulata
Craib.
J
Thai
Tradit
Altern
Med
2009;
7:
146-‐155
[5] http://www.chemspider.com/Chemical-‐Structure.10305920.html
[6] Cushnie
TP,
Lamb
AJ.
Antimicrobial
activity
of
flavonoids.
Int
J
Antimicrob
Agents
2005;
26:
343-‐356
P410
Evolution
with
age
of
main
bitter
compounds
in
the
roots
of
culti-‐
vated
Gentiana
lutea
subsp.
aurantiaca
Óscar
González-‐López,
Sara
Mayo,
Álvaro
Rodríguez-‐González,
Carro,
Guzmán,
Paulo
H.
da
Silva,
Pedro
A.
Casquero
Research
Group
of
Engineering
and
Sustainable
Agriculture,
Natural
Resources
Institute,
University
of
León,
Av.
Portugal
41,
24071
León,
Spain
Gentiana
lutea
L.
subsp.
aurantiaca
is
an
endemic
plant
distributed
in
the
northwest
part
of
the
Iberian
Peninsula
which
shows
clear
genetic
differences
compared
to
other
subspecies
of
G.
lutea
[1]
.
This
subspecies
has
been
widely
used
in
traditional
medicine,
especially
due
to
the
high
concentration
of
bitter
compounds
in
its
roots,
mainly
secoiridoid
glycosides.
The
concentration
of
bitter
compounds
in
this
subspecies
is
similar
compared
with
other
subspe-‐
cies
as
lutea
or
vardjanii
[2].
Like
other
G.
lutea
L.
subspecies,
aurantiaca
subspecies
is
endan-‐
gered
because
of
intensive
and
illegal
collection.
The
cultivation
of
G.
lutea
subsp.
aurantiaca
could
allow
the
protection
of
this
subspecies
and
produce
roots
with
higher
concentration
in
bitter
compounds.
In
this
study
we
compared
the
content
in
the
bitter
compounds
amarogentin,
gentiopicroside,
sweroside
and
swertiamarin
in
the
three,
four
and
five
years
old
roots
of
cultivated
G.
lutea
subsp.
aurantiaca
collected
in
the
autumn
of
2010,
2011
and
2012
with.
The
experimental
field
was
located
near
Leon
(Spain).
For
this
purpose,
methanolic
extracts
of
complete
root
systems
were
analysed
by
HPLC.
The
results
(Error!
Reference
source
not
found.)
show
that
the
concentration
of
amarogentin
decreases
significantly
with
plant
age.
However,
the
concentration
of
swertiamarin
is
significantly
higher
for
older
root
systems,
as
is
the
dry
weight,
which
shows
a
significant
increase
between
years
four
and
five.
Gentiopicroside
and
sweroside
concentrations
did
not
show
a
homogeneous
evolution
or
significant
differences.
The
evolution
of
the
bitter
compounds
regarding
the
age
of
the
roots
is
variable,
favoring
col-‐
lecting
younger
roots
if
the
desired
bitter
compound
is
amarogentin,
due
to
its
high
bitter
in-‐
dex
[3].
Conversely,
older
plants
yield
a
higher
concentration
of
swertiamarine
and
root
mass.
Table
1.
Dry
weight
(g)
and
bitter
compound
concentration
(mg/g
of
root
dry
weight)
in
3,
4
and
5
years
old
roots
systems
of
cultivated
G.
lutea
L.
subsp.
aurantiaca
.
Bars
with
different
letters
for
the
dry
weight
and
every
bitter
compound
are
significantly
different
(P
≤
0.05)
according
to
according
to
LSD.
Age Dry weight Amarogentin Gentiopicroside Sweroside Swertiamarin
(years) (g) (mg/g) (mg/g) (mg/g) (mg/g)
3 4.79 b 0.099 a 6.53 ab 0.297 a 0.309 b
4 9.59 b 0.084 b 7.05 a 0.373 a 0.380 a
5 16.73 a 0.071 b 6.16 b 0.297 a 0.367 a
Acknowledgements:
Regional
Ministry
of
Education
of
the
Junta
de
Castilla
y
León
and
the
European
So-‐
cial
Fund
PIRTU
grants.Orden
EDU/1867/2009.
And
Regional
Ministry
of
the
Enviroment
of
the
Junta
de
Castilla
y
León.
Project
(2008/00134/001)
Keywords: Traditional medicine, medicinal plants, Gentiana
References:
[1] González-‐López
O,
Polanco
C,
György
Z,
Pedryc
A,
Casquero
PA..
Genetic
variation
of
the
endangered
Gentiana
lutea
L.
var.
aurantiaca
(Gentianaceae)
in
populations
from
the
northwest
Iberian
Peninsula.
Int
J
Mol
Sci
2014;
10052-‐10066
[2] González-‐López
O,
Carro
G,
Aiello
N,
Scartezzini
F,
Casquero
PA.
Main
bitter
compounds
of
Gentiana
lutea
var.
aurantiaca
roots
wild
collected
in
the
Leon
Province
(Spain).
Planta
Med
2014;
80:
1489-‐1490
[3] Ando
H,
Hirai
Y,
Fujii
M,
Hori
Y,
Fukumura
M,
Niiho
Y,
Nakajima
Y,
Shibata
T,
Toriizuka
K,
Ida
Y.
The
chemical
constituents
of
fresh
Gentian
root.
J
Nat
Med
2007;
61:
269-‐279
P411
Antiproliferative
diterpenoids
from
the
roots
of
Podocarpus
ne-‐
riifolius
P.
Annécie
Benatrehina1,
Tran
Ngoc
Ninh2,
Carla
Slebodnick3,
Djaja
Djendoel
Soejarto4,5,
L.
Harinantenaina
Rakotondraibe1,
A.
Douglas
Kinghorn1
1
Division
of
Medicinal
Chemistry
and
Pharmacognosy,
College
of
Pharmacy,
The
Ohio
State
University,
500
W.
12th
Ave,
Columbus,
Ohio
43210,
United
States,
2
Institute
of
Ecology
and
Biological
Resources,
Vietnam
academy
of
science
and
Technology,
Hoang
Quoc
Viet,
Cau
Giay,
Hanoi,
Vietnam,
3
Department
of
Chemistry
and
Virginia
Tech
Center
for
Drug
Discovery,
M/C
0212,
Virginia
Tech,
1981
Kraft
Drive,
Blacksburg,
Virginia
24061,
United
States,
4
Department
of
Medicinal
Chemistry
and
Pharmacognosy,
College
of
Pharmacy,
University
of
Illinois
at
Chicago,
833
S.
Wood
Street,
Chicago,
Illinois
60612,
United
States,
5Science
and
Education,
Field
Museum
of
Natural
History,
1400
S.
Lake
Shore
Dr.,
Chicago,
Illinois
60605,
United
States
Plant
samples
collected
in
the
Southeast
Asian
rainforests
were
subjected
to
initial
screening
against
the
HT-‐29
human
colon
cancer
cell
line,
and
Podocarpus
neriifolius
D.
Don
(Podocar-‐
paceae),
obtained
from
Vietnam,
with
an
IC50
value
of
8.2
µg/mL,
was
selected
for
further
in-‐
vestigation
aiming
for
the
discovery
of
potential
anticancer
lead
agents.
Bioactivity-‐guided
purification
of
the
active
ethyl
acetate
fraction
(IC50
=
4.3
µg/mL)
from
the
dried
root
sample
of
P.
neriifolius
has
so
far
afforded
three
type-‐B
podolactones,
namely,
makilactones
E
(1)
and
G
(2)
and
inumakilactone
A
(3),
all
characterized
by
the
presence
of
a
7α,8α-epoxy-9(11)-
enolide functionality [1-3]. Their
structures
were
determined
by
interpretation
of
spectroscop-‐
ic
data
and
comparison
with
the
existing
literature
[1,2].
In
addition,
the
X-‐ray
crystallography
data
of
1
was
obtained
to
confirm
its
absolute
configuration.
While
3
has
shown
termiticidal
and
cytotoxic
activity
against
murine
leukemia
cells
P388
in
vitro
[1,2],
in
this
study,
it
exhib-‐
ited
potent
activity
against
HT-‐29
with
an
IC50
value
of
1.1
µM.
On
the
other
hand,
compounds
1
and
2
were
non-‐cytotoxic
in
this
assay.
In
this
presentation,
the
isolation
and
structure
de-‐
termination
of
these
compounds
are
described
as
well
as
the
dereplication
of
additional
active
sub-‐fractions,
which
suggests
the
presence
of
different
active
podolactones
and
totarane-‐type
diterpenes.
Acknowledgements:
Financial
Support
for
this
project
was
provided
by
the
U.S.
National
Cancer
Institute
(NCI),
National
Institute
of
Health
(NIH)
through
a
program
project
2P01
CA125066
awarded
to
Prof.
A.
Douglas
Kinghorn
Keywords:
Podocarpus,
neriifolius,
podolactones,
HT-‐29
References:
[1] Sato,
K.,
Inaba
Y,
Park
HS,
Akiyama
T,
Koyama
T,
Fukaya
H,
Aoyagi
Y,
Takeya
K.
Cytotoxic
bisnor-‐
and
norditerpene
dilactones
having
7α,8α-‐epoxy-‐9(11)-‐enolide
substructure
from
Podocarpus
macrophyllus
D.
Don.
Chem
Pharm
Bull
2009;
57:
668-‐679
[2] Ito
S,
Kodama
M,
Sunagawa
M,
Takahashi
T,
Imamura
H,
Honda
O.
Structure
of
inumakilactone
A,
a
bisnorditerpenoid.
Tetrahedron
Lett
1968;
17:
2065-‐2070
[3] Barrero
AF,
Quilez
Del
Moral
JF,
Mar
Herrador
M.
Podolactones:
a
group
of
biologically
active
norditerpenoids.
Stud
Nat
Prod
Chem
2003;
28:
453-‐516
P412
Rhizome
weight
and
bioactive
compounds
of
Dioscorea
birmanica
under
different
shading
Panumart
Rithichai1,
Yaowapha
Jirakiattikul1,
Piyapat
Khemwichai1,
Srisopa
Ruangnoo2
and
Arunporn
Itharat2
1Department
of
Agricultural
Technology,
Faculty
of
Science
and
Technology,
Thammasat
University,
Rangsit
Campus,
Pathumthani
12120,
Thailand;
2Department
of
Applied
Thai
Traditional
Medicine,
Fac-‐
ulty
of
Medicine,
Thammasat
University,
Rangsit
Campus,
Pathumthani
12120,
Thailand
The
rhizomes
of
the
medicinal
plant
Dioscorea
birmanica
Prain
&
Burkill
[Dioscoreaceae]
are
used
in
Thai
traditional
medicine
for
the
treatment
of
cancer.
The
ethanolic
extract
of
D.
bir-‐
manica
rhizome
exhibited
high
cytotoxic
activity
against
lung,
colon
and
breast
cancers
[1].
Diosgenin
3-‐O-‐α-‐L-‐rhamnopyranosyl
(1→2)-‐β–D-‐glucopyranoside
or
prosapogenin
A
of
dios-‐
cin
(DBS1)
isolated
from
D.
birmanica
rhizome
showed
high
cytotoxic
activity
against
lung
cancer
but
was
less
toxic
against
normal
lung
cells
[2].
The
rhizomes
are
typically
collected
from
the
forest
and
cultivation
methods
to
produce
rhizomes
for
medicinal
use
are
still
lim-‐
ited.
Therefore,
rhizome
weight
and
bioactive
compounds
of
D.
birmanica
under
different
shading
were
examined.
The
plants
were
grown
in
plastic
pots
under
0%,
50%
and
70%
shad-‐
ing
and
the
rhizomes
were
harvested
at
18
months
after
planting.
The
50%
shading
revealed
the
significantly
highest
dry
rhizome
weight
per
plant
specimen
275.41
±
97.05
g
and
total
phenolic
compounds
as
38.18
±
5.38
mg
gallic
acid
equivalent/g
dry
extract.
DBS1
content
and
EC50
of
DPPH
radical
scavenging
capacity
were
not
significantly
different
among
shading
treatments
as
2.38
±
1.44–3.58
±
1.91%
w/w
and
22.63
±
6.39–36.15
±
14.06
µg/ml,
respec-‐
tively.
Based
on
these
results,
D.
birmanica
should
be
cultivated
under
50%
shading.
Acknowledgements:
This
work
was
supported
by
the
Higher
Education
Research
Promotion
and
National
Research
University
Project
of
Thailand,
the
Office
of
the
Higher
Education
Commission
Keywords:
Dioscorea
birmanica,
dioscin,
DPPH,
phenolic
compound,
yield
References:
[1] Itharat
A,
Houghton
PJ,
Eno-‐Amooquaye
E,
Burke
PJ,
Sampson
JH,
Raman
A.
In
vitro
cytotox-‐
ic
activity
of
Thai
medicinal
plants
used
traditionally
to
treat
cancer.
J
Ethnopharmacol
2004;
90:
33−38.
[2] Jaiaree
N,
Itharat
A,
Kumapava
K.
Cytotoxic
saponin
against
lung
cancer
cells
from
Di-‐
oscorea
birmanica
Prain
&
Burkill.
J
Med
Assoc
Thai
2010;
93:
192−197
P413
Antiplasmodial
activity
and
identification
of
a
new
iridoid
tri-‐
acetate
from
Heinsia
crinata
Tshisekedi
T.
P.1,2,
Kalenda
T.
D.2,
Mutwale
K.
P.1,2,
Cieckiewicz
E.1,
Jansen
O.1,
Angenot
L.1,
Tits
M.1,
Frédérich
M.1
1
University
of
Liege
(ULg),
Department
of
Pharmacy,
CIRM,
Laboratory
of
Pharmacognosy,
CHU,
B36,
4000
Liege,
Belgium,
2
University
of
Kinshasa
(UNIKIN),
Faculty
of
Pharmacy,
Department
of
Pharma-‐
cognosy
and
medicinal
Chemistry,
127
Kinshasa
XI,
RD.
Congo
Malaria
is
a
serious
public
health
challenge
and
one
of
the
biggest
impediments
to
global
de-‐
velopment.
Investigations
were
conducted
on
Heinsia
crinata
(Afz)
G.
Taylor
(Rubiaceae)
which
is
a
shrub
with
woody
stems
and
branches,
collected
from
DRC’s
Botanical
Gardens,
on
July
and
September
2013.
The
aim
of
this
work
was
the
evaluation
of
the
in
vitro
and
in
vivo
antiplasmodial
activity
of
the
stem
bark
extracts,
investigation
of
the
antioxidant
activity
of
methanolic
extracts,
and
finally
the
bioguided
fractionation.
Culture
of
P.
Falciparum
chloro-‐
quine-‐sensitive
(3D7)
was
maintained
as
described
by
Frédérich
et
al.
[1].
The
inhibition
of
the
parasite
growth
was
evaluated
by
colorimetric
method
(p-‐LDH
assay)
[2].
Ethanolic
ex-‐
tract
was
tested
in
vivo
at
the
concentration
of
300
mg/Kg
per
os,
using
a
protocol
based
on
the
4-‐day
suppressive
test
[3].
ABTS
assay
based
on
the
method
reported
by
Franck
et
al.
[4]
and
DPPH
radical
scavenging
capacity
according
to
the
method
developed
by
Brand-‐Williams
et
al.
[5],
were
assessed.
The
results
confirmed
the
antiplasmodial
potential
of
the
extracts
with
IC50
value
of
29.2
±
1.39
µg/mL.
The
plant
exhibited
a
moderate
in
vivo
antiplasmodial
activity
in
Plasmodium
berghei-‐infected
mice
with
48.5
%
of
inhibition
of
the
parasite
growth,
a
good
antioxidant
activity
with
IC50
values
of
66.22
±
2.68
µg
/
mL
in
ABTS
assay
and
234.9
±
41.46
µg/mL
in
DPPH
assay.
Finally,
an
in
vitro
bioguided
fractionation
using
preparative
HPLC
and
TLC
techniques,
allowed
the
identification
of
the
new
iridoid,
Lamiridoside
tri-‐
acetate
by
mean
of
UV,
MS,
and
NMR
spectroscopic
analysis.
This
compound
exhibited
a
mod-‐
erate
in
vitro
antiplasmodial
activity,
with
an
IC50
value
of
16.39
±
0.43
µg/mL.
Acknowledgements: The University of Liege, laboratory of Pharmacognosy.
Keywords: Malaria, antiplasmodial, Heinsia crinata, lamiridoside tri-‐acetate
References:
[1] Frédérich
M,
Jacquier
M.
J,
Thépenier
P,
De
Mol
P,
Tits
M,
Philippe
G,
Delaude
C,
Angenot
L,
and
Zèches-‐Hanrot
M.
Antiplasmodial
activity
of
alkaloids
from
various
Strychnos
species,
J
Nat
Prod
2002;
65:
1381-‐1386
[2] Makler
M.
T,
Ries
J.
M,
Williams
J.
A,
Bancroft
J.
E,
Piper
R.
C,
Gibbins
B.
L,
and
Hinrichs
D.
J.
Parasite
lactate
dehydrogenase
as
an
assay
for
Plasmodium
falciparum
drug
sensitivity,
Am
J
Trop
Med
Hyg
1993;
48:
739-‐741
[3] Fidock
D.
A,
Rosenthal
P.
J,
Croft
S.
L,
Brun
R,
Nwaka
S,
and
Einstein
A.
Antimalarial
drug
discovery:
efficacy
models
for
compound
screening,
Nat
Rev
Discov
2004;
3:
509-‐520
[4] Franck
T,
Mouithys-‐mickalad
A,
Robert
T,
Ghitti
G,
Deby-‐dupont
G,
Neven
P,
Serteyn
D.
Chemico-‐biological
interactions
differentiation
between
stoichiometric
and
anticatalytic
antioxidant
properties
of
benzoic
acid
analogues :
A
structure/redox
potential
relationship
study.
Chem
Biol
Interact
2013;
206:
194-‐203
[5] Brand-‐Williams
W,
Cuvelier
ME,
Berset
C.
Use
of
a
free
radical
method
to
evaluate
antioxidant
activity.
LWT
-‐
Food
Sci
Technol
1995;
28:
25-‐30
P414
Evaluation
of
in
vitro
activity
and
ultrastructural
changes
in
Leishmania
amazonensis
caused
by
sesquiterpene
lactones
from
Calea
pinnatifida
(Asteraceae)
Patricia
Sartorelli1,
Meire
L.
Yoshinaga1,
Marcelo
José
P.
Ferreira2,
João
Henrique
G.
Lago1,
Luiz
Felipe
D.
Passero3
1
Institute
of
Environmental,
Chemical
and
Pharmaceutical
Sciences,
Federal
University
of
São
Paulo,
São
Paulo,
Brazil,
2
Institute
of
Biosciences,
University
of
São
Paulo,
Brazil,
3
Faculty
of
Medicine,
University
of
São
Paulo,
Brazil
Calea
genus,
belonging
to
the
Asteraceae,
is
composed
by
approximately
125
species.
Some
of
these
species
were
chemically
investigated
and
sesquiterpene
lactones
as
well
as
furan
ses-‐
quiterpenes
have
been
identified
[1].
There
are
several
biological
activities
related
to
the
compounds
present
in
the
genus
Calea,
such
as
anti-‐inflammatory,
antiplasmodial,
antifungal,
antimicrobial
and
cytotoxic,
which
are
credited
to
the
presence
of
sesquiterpene
lactones,
mainly
germacranolide
derivatives
[2].
C.
pinnatifida,
popularly
known
as
"aruca"
and
“cipó-‐
cruz”,
has
been
encountered
specially
in
Brazilian
Cerrado
biome
(“savanna
like”)
and
has
been
used
in
folk
medicine
to
treat
stomachaches,
giardiasis
and
amebiasis.
In
Brazil
this
spe-‐
cies
is
commercially
available
in
popular
market
as
an
ethanol
extract
of
the
leaves
to
treat
amoebic
dysentery.
Chemically,
the
aerial
parts
are
composed
of
fatty
esters,
glycoside
of
p-‐
hydroxybenzoic
acid,
anisic
acid,
sitosterol,
stigmasterol,
polyacetylene
derivatives
and
ger-‐
macranolides
[3].
Previous
studies
revealed
germacranolides
displayed
cytotoxic
potential
inducing
apoptosis
in
different
tumor
cells,
as
well
as
displayed
inhibitory
activity
of
NF-‐ĸB
factor
[4].
In
present
work,
the
bioactivity
guided
fractionation
of
crude
ethanol
extract
from
leaves
of
C.
pinnatifida
led
to
isolation
of
two
sesquiterpene
lactones:
arucanolide
and
calealactone
C,
which
structures
were
established
on
the
basis
of
spectroscopic
analysis.
The-‐
se
compounds
displayed
potent
activity
against
promastigote
forms
of
Leishmania
(L.)
ama-‐
zonensis
with
EC50
of
1.7
and
4.6
µg/mL
to
arucanolide
and
calealactone
C,
respectively.
These
compounds
caused
ultrastructural
changes
in
L.
amazonensis
promastigotes
leading
to
a
loss
of
their
classical
structural
morphology,
as
evidenced
by
electron
microscopy.
Additionally,
calealactone
C
presented
low
cytotoxicity
for
J774
macrophages
(CC50
31.73
µg/mL),
similar
to
positive
controls
and
presented
antiamastigote
activity
with
EC50
of
4.24
µg/mL.
Acknowledgements:
The
authors
would
like
to
thank
FAPESP
and
CNPq
for
providing
financial
support
to
this
study.
Keywords: Calea pinnatifida, sesquiterpene lactones, arucanolide, calealactone C, antileish-‐
manial activity, ultrastructural changes
References:
[1] Seaman
FC.
Sesquiterpene
lactones
as
taxonomic
characters
in
the
Asteraceae.
Bot
Rev
1982;
48:
121-‐594
[2] Nakagawa
Y,
Iinuma
M,
Matsuura
N,
Yi
K,
Naoi
M,
Nakayama
T,
Nozawa
Y,
Akao
J.
Phar-‐
macol
Sci
2005;
97:
242-‐252
[3] Ferreira
ZS,
Roque
NF,
Gottlieb
OR,
Oliveira
F,
Gottlieb
HE.
Structural
clarification
of
ger-‐
macranolides
from
Calea
species.
Phytochemistry
1980;
19:
1481-‐1484
[4] Rivero
A,
Quintana
J,
Eiroa
JL,
López
M,
Triana
J,
Bermejo
J,
Estévez
F.
Potent
induction
of
apoptosis
by
germacranolide
sesquiterpene
lactones
on
human
myeloid
leukemia
cells.
Eur
J
Pharmacol
2003;
482:
77-‐84
P415
Endemic
Asteraceae
from
Madeira
archipelago:
A
relation
of
hy-‐
poglycemic
activity
to
their
polyphenolic
composition
Vítor
Spínola1,
Sandra
Gouveia-‐Figueira2,
Paula
C.
Castilho1
1
Centro
de
Química
da
Madeira
(CQM),
Madeira
University,
Campus
Penteada,
9000-‐390
Funchal,
Portu-‐
gal,
2
Department
of
Chemistry,
Umeå
University,
901
87
Umeå,
Sweden
The
vascular
flora
of
Madeira
archipelago
is
exuberant
and
diverse,
comprising
over
1220
species
of
which
10%
are
endemic
[1].
In
this
work,
nine
endemic
Asteraceae
plant
species
were
studied
for
their
phenolic
profile
and
anti-‐hyperglycemic
effects.
The
interest
and
selec-‐
tion
of
these
species
are
related
to
their
use
in
folk
medicine.
Two
main
groups
of
plants
were
analyzed:
Helichrysum
subspecies
(H.
devium
Johns,
H.
melaleucum
Rchb.
ex.
Holl,
H.
monizii
Lowe
and
H.
obconicum
DC.)
and
other
five
endemic
species:
Argyranthemum
pinnati-‐
fidum
Lowe,
Artemisia
argentea
L’Her,
Calendula
maderensis
DC,
Cynara
cardunculus
L.
var.
ferocissima
and
Phagnalon
lowei
DC.
Alcoholic
extracts
were
evaluated
for
their
inhibitory
activity
towards
key
enzymes
linked
to
type
2
diabetes
using
in
vitro
assays.
Six
extracts
showed
stronger
inhibitory
efficacy
[IC50
0.57
to
1.35
mg/mL]
than
commercial
drug
acarbose
(positive
control,
IC50
1.62
mg/mL)
towards
α-‐glucosidase;
exceptions
were
H.
monizii,
C.
ma-‐
derensis
and
C.
cardunculus.
All
extracts
were
less
active
than
the
control
towards
α-‐amylase
(IC50
ranging
from
1.5
to
9.1
mg/mL
versus
0.02
of
acarbose).
Phenolic
profiles
were
obtained
by
HPLC-‐MSn
and
caffeoylquinic
acid
derivatives
(esterified
with
several
acyl
groups)
were
found
as
the
main
compounds
in
all
plant
species
under
analysis,
ranging
from
86
(A.
glandu-‐
losa
leaves)
to
1918
(P.
lowei
leaves)
mg/100g
of
dry
plant.
The
anti-‐diabetic
properties
of
these
caffeoylquinic
acid
derivatives
have
been
extensively
reported
[2,
3].
Four
out
of
ten
studied
species
(A.
argentea,
A.
pinnatifidum,
H.
melaleucum
and
P.
lowei)
are
potent
inhibitors
of
α-‐glucosidase
with
moderate
lipase
and
low
α-‐amylase
inhibition,
which
is
ideal
for
pre-‐
venting
bacterial
fermentation
of
excessive
indigested
carbohydrates
in
the
colon.
Acknowledgements:
Thanks
are
due
to
Fundação
para
a
Ciência
e
a
Tecnologia
(FCT,
Portugal)
for
Vítor
Spínola’s
Ph.D.
grant
(SFRH/BD/84672/2012,
for
Project
PEst-‐OE/QUI/UI0674/2014
and
to
the
Portu-‐
guese
National
Mass
Spectrometry
Network
(RNEM2014)
in
the
framework
of
the
National
Programme
for
Scientific
Re-‐equipment,
with
funds
from
POCI
2010
(FEDER)
Keywords:
Asteraceae,
enzyme
inhibition,
phenolic
profiles
References:
[1]
Rivera
D,
Óbon
C.
The
ethnopharmacology
of
Madeira
and
Porto
Santo
Islands,
a
review.
J
Ethnopharmacol
199;
46:
73-‐93
[2]
McCarty
MF.
Nutraceutical
resources
for
diabetes
prevention-‐an
update.
Med
Hypotheses
2005;
64:
151-‐158
[3]
El-‐Abhar
HS,
Schaalan
MF.
Phytotherapy
in
diabetes:
Review
on
potential
mechanistic
per-‐
spectives.
World
J
Diabetes
2014;
5:
176-‐197
P416
4-‐Methyl-‐1-‐alkylresorcinols,
novel
chemotaxonomic
markers
from
the
Spanish
medicinal
plant
Mercurialis
tomentosa
L.
Peter
Lorenz,
Miriam
Heinrich,
Dietmar
R.
Kammerer,
Florian
C.
Stintzing
WALA
Heilmittel
GmbH,
Department
of
Analytical
Development
&
Research,
Section
Phytochemical
R e-‐
search,
Dorfstr.
1,
D-‐73087
Bad
Boll/Eckwaelden,
Germany
Mercurialis
tomentosa
is
a
medicinal
plant,
the
extracts
of
which
have
recently
been
demon-‐
strated
to
exert
PGE2
and
TNF-‐α
inhibitory
activity
[1].
However,
phytochemical
data
to
which
these
pharmacological
effects
are
related
were
not
provided.
Consequently,
more
detailed
studies
were
performed
comprising
GC-‐MS
and
LC-‐MSn
investigations
of
CHCl3
extracts
from
M.
tomentosa,
yielding
several
homologous
alkylresorcinols.
For
structure
assignment
a
novel
facile
synthesis
of
one
representative
(4-‐Me-‐AR-‐C19:0)
was
developed,
starting
with
a
Grig-‐
nard
reaction.
The
resulting
reaction
product
was
identical
to
the
natural
product
in
terms
of
its
chromatographic
and
spectroscopic
features.
In
contrast
to
other
Mercurialis
species
(M.
perennis,
M.
annua)
which
exclusively
contain
1-‐alkylresorcinol
homologues
[2],
4-‐methyl-‐1-‐
alkylresorcinols
(side
chain
lengths:
C15-‐C25)
are
predominant
in
M.
tomentosa.
Furthermore,
4-‐Me-‐AR-‐C19:0
(IC50
=
37.8
µM)
exhibited
promising
DPPH
free
radical
scavenging
activity
when
compared
to
trolox
(IC50
=
21.0
µM)
–
substantiating
the
antioxidant
features
of
these
amphiphilic
molecules.
The
compounds
characterized
in
this
study
may
be
utilized
for
species
differentiation
based
on
fingerprinting
methods.
50
45
40
Relative
peak area [%]
35
30
25
20
15
10
0
C15 C17 C18 C19 C20 C21 C22 C23
M.
tomentosa L.
Side
chain length
Keywords:
Dog's
mercury,
Euphorbiaceae,
alkylresorcinols
References:
[1] Bremner
P,
Rivera
D,
Calzado
MA,
Obón
C,
Inocencio
C,
Beckwith
C,
Fiebich
BL,
Munoz
E,
Heinrich
M.
Assessing
medicinal
plants
from
south-‐eastern
Spain
for
potential
anti-‐
inflammatory effects targeting nuclear factor-‐kappa B and other pro-‐inflammatory media-‐
tors. J Ethnopharmacol 2009; 124: 295-‐305
[2] Lorenz P, Duckstein S, Conrad J, Knödler M, Stintzing FC. An approach to the chemotaxo-‐
nomic differentiation of two European dog’s mercury species: Mercurialis annua L. and M.
perennis L. Chem Biodivers 2012; 9: 282-‐297
P417
Assessment
of
Rauvolfia
nukuhivensis:
from
uses
to
biological
ac-‐
tivities
and
chemodiversity
Nicolas
Martin1,
Sara
F.
Ferreiro2,
Eva
Alonso-‐Lopes2,
Soizic
Prado3,
Florent
Barbault4,
Maël
Nicolas5,
Gaël
Lecellier6,
Christian
Paetz7,
Marc
Gaysinski5,
Olivier
Thomas5,
Luis
Botana2,
Phi-‐
la
Raharivelomanana1
1
University
of
French
Polynesia,
EIO
UMR
241,
Faa'a,
French
Polynesia,
2
Departimento
de
Farmacología,
Facultad
de
Veterinaria,
Universidad
de
Santiago
de
Compostela,
Lugo,
Spain,
3
MNHN
UMR
CMAM
7245
Paris,
France,
4
Laboratoire
ITODYS
UMR
CNRS
7086,
Université
Paris
Diderot,
Sorbonne
Paris
Cité,
15
rue
J.-‐A.
de
Baïf,
75013
Paris,
France,
5
Geoazur,
UMR
7329,
Université
de
Nice
–
Sophia
Antipolis,
250
rue
Albert
Einstein,
Sophia
Antipolis,
06560
Valbonne,
France,
6
Université
de
Versailles
Saint-‐Quentin
en
Yvelines,
55
Avenue
de
Paris,
78000
Versailles,
France,
7
Max
Planck
Institute
for
Chemical
Ecology,
NMR
department,
Jena,
Germany
Rauvolfia
genus,
biogeographically
widespread
in
tropical
areas,
includes
many
species
used
in
traditional
medicine.
Studies
of
these
species
showed
that
their
uses
are
related
to
their
bioactive
content
mostly
composed
by
alkaloid
compounds.
Rauvolfia
nukuhivensis,
called
lo-‐
cally
“tueiao”,
is
an
endemic
species
grown
in
Nuku-‐Hiva
island
located
in
Marquesas
archi-‐
pelago
where
the
bark
is
used
for
intimate
woman
care
[1].
Ethnopharmacological
approach
was
adopted
to
assess
this
plant.
For
that
purpose,
firstly,
we
checked
if
the
plant
extract
played
an
antiseptic
role
within
antimicrobial
effects
by
testing
the
extract
and
contents
on
bacterial
and
fungi
strains
(Escherichia
coli,
Staphylococcus
aureus,
Candida
albicans
and
As-‐
pergillus
niger)
and
bioassay
results
showed
moderate
to
low
activities.
Then,
we
supposed
that
traditional
medicinal
uses
of
this
plant
may
be
linked
to
biological
activities
having
close
relationship
with
secretory
mechanisms
regulation.
This
hypothesis
was
investigated
by
the
ion
channels
inhibition
related
to
osmotic
exchanges
and
then,
inhibiting
effects
on
hNav1.6
currents
and
hERG
channel
were
so
investigated.
Some
alkaloidal
constituents,
and
more
es-‐
pecially
nukuhivensiums,
were
shown
to
significantly
induce
a
reduction
of
IKr
amplitude
(hERG
current).
According
to
these
biological
activities,
a
computational
study
through
dock-‐
ing
was
performed
in
order
to
illustrate
these
results
[2,3].
The
chemodiversity
of
the
alka-‐
loids
from
the
bark
of
this
plant
was
assessed
by
three
inte-‐grated
approaches
mainly:
struc-‐
tural
elucidation,
phylogenetic
analysis
and
a
putative
biosynthesis
hypothesis
statement.
Structural
elucidation
by
spectroscopic
means
(MS,
NMR)
led
to
the
identification
of
13
major
constituents
belonging
to
four
distinct
indole
alkaloid
skeletons
(ajmalane,
sarpagane,
macro-‐
line
and
β-‐carboline)
within
six
new
naturally
occuring
compounds.
As
the
Rauvolfia
genus
is
well
distributed
in
the
Pacific
region,
we
carried
out
a
phylogenetic
study
(using
DNA
barcod-‐
ing
method)
of
Rauvolfia
species,
endemic
or
indigenous
in
Oceania,
as
a
second
approach,
aiming
to
a
better
understanding
of
the
occurence
and
distribution
of
these
alkaloids.
The
very
rare
co-‐occurence
of
these
alkaloids
belonging
to
four
different
skeletons
inspired
to
set
up
a
putative
biosynthesis
of
these
components,
as
a
third
way
of
investigation
of
their
che-‐
modiversity
[2-‐4].
Acknowledgements:
The
‘‘Ministère
de
l’Enseignement
Supérieur
et
de
la
Recherche’’
of
the
French
gov-‐
ernment
is
acknowledged
for
providing
PhD
grant.
We
are
thankful
to
J.F.
Butaud
and
to
French
Polyne-‐
sia
DIREN
department
for
sample
collection,
plant
material
identification
and
helpful
discussions.
The
authors
wish
to
thank
J.-‐M.
Guigonis
(Plate-‐Forme
Bernard
Rossi)
for
HRESIMS
measurements.
Keywords: Rauvolfia nukuhivensis, Apocynaceae, ethnopharmacology, indole alkaloids, hERG
channel activity inhibition, Docking calculation
References:
[1] Girardi
C,
Butaud
JF,
Ollier
C,
Ingert
N,
Weniger
B,
Raharivelomanana
P,
Moretti
C.
Herbal
medicine
in
the
Marquesas
islands,
J
Ethnopharmacol
2015;
161:
200-‐213.
[2] Martin
NJ,
Prado
S,
Lecellier
G,
Thomas
O,
Raharivelomanana
P.
Nukuhivensiums,
indolo
[2,3-‐α]
quinoliziniums
from
the
Marquesan
plant
Rauvolfia
nukuhivensis.
Molecules
2012;
17:
12015-‐12022.
[3] Martin
NJ,
Ferreiro
SF,
Barbault
F,
Nicolas
M,
Lecellier
G,
Paetz
C,
Gaysinski
M,
Alonso
E,
Thomas
OP,
Botana
LM,
Raharivelomanana
P.
Indole
alkaloids
from
the
Marquesan
plant
Rauvolfia
nukuhivensis
and
their
effects
on
ion
channels.
Phytochemistry
2015;
109:
84-‐
65.
[4] Martin
NJ,
Nicolas
M,
Lecellier
G,
Raharivelomanana
P.
Isolation
and
characterization
pro-‐
cedure
for
indole
alkaloids
from
the
Marquesan
plant
Rauvolfia
nukuhivensis,
Bio-‐protocol
2015;
5:
e1625.
P418
Anti-‐ageing
activity
of
Fitchia
nutans
extract,
a
Polynesian
tradi-‐
tional
monoï
skin
care
ingredient
Jean-‐Luc
Ansel1,2,
Quoc
Ly1,
Jean-‐François
Butaud3,
Mael
Nicolas1,4,
Gaëtan
Herbette5,
Laurent
Peno-‐Mazzarino6,
Elian
Lati6,
Phila
Raharivelomanana1
1
UMR
241
EIO
Université
de
la
Polynésie
Française,
Tahiti,
98702
Faa’a,
Polynésie
Française,
2
Cosmetic-‐
Valley,
1
Place
de
la
cathédrale,
28200
Chartres,
France,
3
Consultant
en
foresterie
et
botanique
polyné-‐
sienne,
BP
52832,
98716
Pirae,
Tahiti,
Polynésie
Française,
4
Geoazur,
UMR
7329,
Université
de
Nice
–
Sophia
Antipolis,
250
rue
Albert
Einstein,
Sophia
Antipolis,
06560
Valbonne,
France,
5
Spectropole,
FR
1739,
Université
Aix
Marseille,
Campus
Saint
Jérôme,
13397
Marseille
cedex
20,
France,
6
Laboratoire
BIO-‐
EC,
1
chemin
de
Saulxier,
91160
Longjumeau,
France
Fitchia
nutans
Hook.f.
(Asteraceae),
a
Polynesian
endemic
plant
locally
called
“’anei”,
was
pre-‐
viously
used
as
a
skin
care
ingredient
included
in
a
sacred
traditional
monoï
preparation
in
French
Polynesia
[1].
Such
use
indicated
interesting
skin
care
properties
to
investigate.
For
that
purpose,
leaves
extract
of
F.
nutans
was
tested
on
ex-‐vivo
human
skin
for
anti-‐ageing
ac-‐
tivity
assays
as
ex-‐vivo
systems
could
mirror
the
morphological
and
immunophenotypic
prop-‐
erties
of
the
tissues
[2].
Abdominal
skin
ex-‐vivo
tissues
used
in
this
study
were
collected
from
surgical
procedures.
Follow-‐up
at
histological
level
using
histochemical
and
immunomarking
techniques
were
performed
within
observation
by
high
resolution
electron
microscopy
[3].
Qualitative
and
quantitative
changes
of
dermal
collagens
(I;
III
and
IV)
and
elastin
fibres
were
investigated
for
tested
ex-‐vivo
skins.
The
results
indicate
its
potential
to
stimulate
collagens
and
elastin
dermal
growth.
To
the
best
of
our
understanding,
only
one
species
(F.
speciosa)
in
the
Fitchia
genus
has
been
investigated
for
its
chemical
composition
[4].
We
report
herein,
the
first
phytochemical
study
of
this
plant
using
spectrometric
and
spectroscopic
methods
(MS,
1D
and
2D
NMR
analysis)
for
structure
elucidation
and
showing
that
main
constituents
are
composed
by
sesquiterpenoids
(such
as
costunolide
derivatives
including
a
new
natural
product:
15-‐isovaleroyloxydihydrocostunolide),
phenylpropanoids
(like
butanoic
acid,
3-‐
methyl-‐4,'-‐(1-‐propen-‐1-‐yl)phenyl
ester),
and
phenolic
derivatives
(like
atranorin).
All
identi-‐
fied
components
contained
ester
functions
(mostly
as
isovaleroyl
esters).
The
sesquiterpene
lactone
costunolide
is
well-‐known
for
its
interesting
biological
activities
such
as
an
antifungal
on
dermatophytes
or
as
an
anti-‐inflammatory
agent
[5,
6].
The
presence
of
costunolide
deriv-‐
atives
as
the
main
components
of
F.
nutans
extract
may
contribute
to
the
observed
anti-‐ageing
properties
[7].
Acknowledgements:
Biopolyval
project
and
Cosmetic-‐Valley
are
deeply
acknowledged
for
financial
sup-‐
port.
Keywords:
Fitchia
nutans,
anti-‐ageing
activity,
ex
vivo
human
skin
tests,
collagen,
elastin,
costunolide,
isovaleryl
esters
References:
[1] Pétard
P,
Haere
Po
No
Tahiti
(Ed.),
Plantes
utiles
de
Polynésie
et
raau
Tahiti,
Tahiti,
1986;
303
[2] Khoshnoodi
J,
Pedccenko
JV,
Hudson
BG.
Mammalian
collagen
IV.
Microsc
Res
Tech
2008;
71:
357-‐370
[3] Seite
S,
Zucchi
H,
Septier
D,
Igondjo-‐Tchen
S,
Senni
K,
Godeau
G.
Elastin
changes
during
chronological
and
photo-‐ageing:
the
important
role
of
lysozyme.
J
Eur
Acad
Dermatol
Ve-‐
nereol
2006;
20:
980-‐987
[4] Bohlmann
F,
Zdero
C,
King
RM,
Robinson
H.
New
sesquiterpene
lactones
and
other
constit-‐
uents
from
Fitchia
speciosa.
Phytochemistry
1980;
19:
1141-‐1143
[5] Duraipandiyan
V,
Al-‐Harbi
NA,
Ignacimuthu
S,
Muthukumar
C.
Antimicrobial
activity
of
sesquiterpene
lactones
isolated
from
traditional
medicinal
plant,
Costus
speciosus
(Koenex.Retz.)
Sm.
BMC
Complement
Altern
Med
2012;
12:
12-‐13
[6] Castro
V,
Murillo
R,
Klaas
CA,
Meunier
C,
Mora
G,
Pahl
HL,
Merfort
I.
Inhibition
of
the
tran-‐
scription
factor
NF-‐kappa
B
by
sesquiterpene
lactones
from
Podachaenium
eminens.
Plan-‐
ta
Med
2000;
66:
591-‐595
[7] Ansel
JL,
Ly
Q,
Butaud
JF,
Nicolas
M,
Herbette
G,
Peno-‐Mazzarino
L,
Lati
E,
Rahariveloma-‐
nana
P.
Activité
anti-‐âge
de
l’extrait
de
Fitchia
nutans,
un
ingredient
cosméticeutique
d’un
monoï
traditionnel
polynésien.
Comptes
Rendus
Chimie
2016
;
http://dx.doi.org/10.1016/j.crci.2016.03.005.
P419
Anti-‐fibrotic
effects
of
Silimarit®
dry
extract
in
the
STAM™
model
of
non-‐alcoholic
steatohepatitis
Philipp Peterburs, Aldo Ammendola, Anja Lechner
Bionorica SE, Kerschensteinerstraße 11-‐15, 92318 Neumarkt, Germany,
Non-‐alcoholic fatty liver disease (NAFLD)/non-‐alcoholic steatohepatitis (NASH) is a major
health concern in western countries with increasing prevalence. Until now, effective therapies
are missing and the most effective way treating NASH/NAFLD is a change in nutrition and life
style. The search for novel therapeutic agents is a challenging problem, because most of the in
vivo models fail to represent the situation in disease progression in humans. Milk thistle, Si-‐
lybum marianum, is a traditionally used medical plant for the treatment of liver injuries. Its
potential in NAFLD/NASH is only poorly characterized. In this study the STAM™ model, a pre-‐
clinically validated animal model, was applied to induce NAFLD/NASH-‐like symptoms in mice.
6 weeks after NAFLD/NASH induction mice received a standardized milk thistle extract (Sili-‐
marit®, 20 and 100mg/kg each day). After 9 weeks body weight, serum parameters, liver
damage, fibrosis, and steatosis were analyzed. Histological analysis of vehicle-‐treated control
animals revealed severe symptoms of NAFLD/NASH which was indicated by fat accumulation
in hepatocytes and fibrosis of liver tissue. Additionally, body weight decreased and serum pa-‐
rameters like blood glucose and triglceride levels increased. Treatment with Silybum maria-‐
num extract had no effect on body weight, blood glucose and triglycerid levels. However, his-‐
tological analysis of sirius red staining showed a marked effect of Silybum marianum extract
on the development of fibrosis. The extract significantly reduced fibrotic areas of the liver
which were stained positive for Sirius red.
In the present study, we demonstrated for the first time the anti-‐fibrotic effect of Silimarit® in
a mouse model for NAFLD/NASH. Mice treated with milk thistle extract showed significantly
decrease in fibrotic areas in the liver. Our results support Silybum marianum extract as a
promising therapic agent for NAFLD/NASH patients.
Acknowledgements: The authors wish to thank Stelic Institute & Co., Tokyo, Japan, for their contributions
to the design and conduct of this study, data analyses and data reporting
Keywords: Milk thistle extract, NASH, fibrosis
P420
In
vitro
and
cellular
antioxidant
activities
of
Antidesma
thwaitesianum
Müll.
Arg.
(Euphorbiaceae)
leaf
extracts
Pintusorn
Hansakul1,3,
Bhanuz
Dechayont2,3,
Nitra
Nuengchamnong4,
Arunporn
Itharat2,3
1
Department
of
Preclinical
Science,
Faculty
of
Medicine,
Thammasat
University,
Pathumthani,
12120,
Thailand,
2
Department
of
Applied
Thai
Traditional,
Faculty
of
Medicine,
Thammasat
University,
Pathumthani,
12120,
Thailand,
3
Center
of
Excellence
in
Applied
Thai
Traditional
Medicine
Research,
Faculty
of
Medicine,
Thammasat
University,
Pathumthani,
12120,
Thailand,
4
Science
Laboratory
Centre,
Faculty
of
Science,
Naresuan
University,
Phitsanulok,
65000,
Thailand.
Fresh
and
fruit
waste
extracts
of
Antidesma
thwaitesianum
Müll.
Arg.
(Euphorbiaceae)
possessed
potent
in
vitro
and
cellular
antioxidant
activities
[1].
Its
leaves
are
edible
as
a
fresh
vegetable,
and
its
leaf
decoc-‐
tion
is
used
to
treat
degenerative
diseases
in
Thai
traditional
medicine.
However,
no
scientific
evidence
supporting
its
health
benefits
has
been
reported.
Accordingly,
we
aimed
to
determine
in
vitro
and
cellular
antioxidant
activities
of
A.
thwaitesianum
leaf
extracts
obtained
from
different
extraction
methods.
The
fresh
leaf
extract
was
prepared
by
blending.
Additionally,
the
dried
leaf
extracts
were
obtained
by
decoc-‐
tion
and
maceration,
including
the
extract
obtained
by
decocting
the
residue
left
after
maceration.
In
chemical
assays,
all
of
the
tested
leaf
extracts
exerted
scavenging
activity
against
DPPH
and
ABTS
radi-‐
cals
(EC50
=
3.54-‐7.46
µg/mL;
6.13-‐11.81
µg/mL).
The
maceration
and
decoction
extracts
possessed
the
highest
antioxidant
activity,
similar
to
the
standard,
Trolox.
In
cell-‐based
assays,
only
the
ethanolic
ex-‐
tract
exerted
the
most
potent
superoxide
(O2.-‐)
and
nitric
oxide
(NO.)
scavenging
activity
(EC50
=
58.12;
71.90
µg/mL).
In
contrast,
the
decoction
extract
possessed
only
strong
NO.
scavenging
action
(EC50
=
91.20
µg/mL).
The
decoction
and
maceration
extracts
contained
high
total
phenolic
and
flavonoid
con-‐
tents,
which
were
partially
correlated
with
their
cellular
antioxidant
activity.
Due
to
its
highest
antioxi-‐
dant
potential,
the
first
comprehensive
identification
of
active
components
of
A.
thwaitesianum
ethanolic
extract
was
performed
by
HPLC-‐ESI-‐QTOF-‐MS.
In
the
analysis,
145
compounds
were
obtained,
as
well
as
14
flavonoids
and
35
phenolic
acids
were
tentatively
identified.
This
is
the
first
report
describing
in
vitro
and
cellular
antioxidant
effects
of
leaf
extracts
of
A.
thwaitesianum,
supporting
the
traditional
use
of
its
leaf
decoction.
Moreover,
its
ethanolic
leaf
extract
has
a
great
potential
as
dietary
supplements.
Acknowledgements:
This
study
was
financially
supported
by
National
Research
University
Project
of
Thailand,
Office
of
the
Higher
Education
Commission,
the
National
Research
Council
of
Thailand
(NRCT),
and
Faculty
of
Medicine,
Thammasat
University.
Keywords:
Antidesma
thwaitesianum
Müll.
Arg.,
cellular
antioxidant
activity,
cell-‐based
as-‐
says
References:
[1] Hansakul
P,
Dechayont,
B
Phuaklee
P,
Prajuabjinda
O,
Juckmeta
T,
Itharat
A.
Cytotoxic
and
antioxidant
activities
of
Antidesma
thwaitesianum
Müll
Arg
(Euphorbiaceae)
fruit
and
fruit waste
extracts.
Trop
J
Pharm
Res,
2015;
14:
627-‐634
P421
Cytotoxic
activity
against
liver
cancer
cell
[HepG2]
of
a
Thai
tradi-‐
tional
remedy
used
for
liver
cancer
treatment
Ponlawat
Maki1,
Thammarat
Tui-‐on2,3
,
Arunporn
Itharat1,2
1
Ph.D
Program
on
Applied
Thai
Traditional
Medicine,
Department
of
Applied
Thai
Traditional
Medicine,
Faculty
of
Medicine,
Thammasat
University,
Klongluang,
Pathumthani
12120,
Thailand
,
2
Department
of
Applied
Thai
Traditional
Medicine,
Faculty
of
Medicine,
Thammasat
University,
Klongluang,
Pathumthani
12120,
Thailand
,
3
Center
of
Excellence
on
Applied
Thai
Traditional
Medicine
Research
(CEATMR),
Thammasat
University,
Klongluang,
Pathumthani
12120,
Thailand
A
Thai
traditional
remedy
of
a
Thai
folk
doctor
has
been
used
to
treat
liver
cancer
for
more
than
30
years.
In
a
preliminary
study
24%
of
liver
cancer
patients
that
used
this
remedy
were
still
alive
after
one
year
[1].
From
the
in-‐depth
interview
of
this
folk
doctor,
we
obtained
the
knowledge
and
principle
of
diagnostic,
treatment
and
the
reason
for
using
medicinal
plants
in
this
cancer
remedy.
The
forty
plants
included
in
this
remedy
were
tested
for
cytotoxic
activity
against
liver
cancer
cells
(HepG2)
by
using
SRB
assay
[2].
The
drug
design
used
the
holistic
principle,
detoxification
of
liver,
renal
and
heart
tonic.
Medicinal
plants
in
this
traditional
Thai
preparation
are
laxative
plants
such
as
Aloe
vera,
Tamarindus
indicus,
and
diuretic
plants
such
as
Ananus
comosus
and
Imperata
cyrindrica.
For
heart
tonic,
he
used
five
flowers
such
as
Jas-‐
minum
sambac,
Nelumbo
nucifera,
Mammea
siamensis,
Mesua
ferrea
and
Mimosop
elengi.
In-‐
geredients
that
the
folk
doctor
believed
had
an
effect
on
the
liver
included
almost
bitter
tast-‐
ing
plants
such
as
Andrographis
paniculata,
Phyllanthus
amarus
and
Solanum
indicum.
Among
the
plants
that
have
been
reported
to
have
anticancer
effect
and
were
the
main
ingredients
in
the
remedy
included
Rhinacanthus
nasuthus
and
Helotropium
indicum.
The
cancer
remedy
was
extracted
by
maceration
with
95%
ethanol
and
boiling
in
water
The
ethanolic
extract
of
this
remedy
showed
high
cytotoxic
activity
against
HepG2
with
IC50
=
18.40
µg/ml
but
the
water
extract
had
no
activity
(IC50
>100
µg/ml).
Thus,
forty
plants
as
ingredients
were
also
macerated
in
95%
ethanol
and
tested
for
cytotoxicity.
Twelve
plants
showed
IC50
less
than
30
µg/ml.
Mammea
siamensis
showed
the
best
cytotoxic
activity
(IC50
=
14.06
±
1.7
µg/ml).
These
results
correlate
with
the
ethnopharmacological
use
of
the
Thai
traditional
remedy.
Acknowledgements:
This
work
was
supported
by
the
National
Research
University
Project
of
Thailand
Office
of
Higher
Education
Commission,
Center
of
Excellence
in
Applied
Thai
Traditional
Medicine,
Facul-‐
ty
of
Medicine,
Thammasat
University
Keywords: Liver cancer, HepG2, SRB assay, cytotoxic activity, Thai Traditional medicine
References:
[1] Marki
P,
Tui-‐On
T,
Itharat
A
The
study
on
wisdom
of
cancer
treatment
by
a
Thai
folk
doc-‐
tor
in
Pechaburi
Province,
Department
of
Applied
Thai
Traditional
Medicine,
Thammasat
University,
2014
[2] Skehan
P,
Storeng
R,
Scudiero
D,
Monks
A,
McMahon
J,
Vistica
D,
Warren
JT,
Bokesch
H,
Kenney
S,
Boyd
MR.
New
colorimetric
cytotoxicity
assay
for
anticancer-‐drug
screening.
J
Natl
Cancer
Inst
1990;
82:
1107-‐1112
P422
Effects
of
biotransformation
of
prenylated
flavonoids
on
antioxi-‐
dative
capacity
Yina
Xiao,
Ik-‐Soo
Lee
College
of
Pharmacy
and
Research
Institute
of
Drug
Development,
Chonnam
National
University,
Gwangju
61186,
Republic
of
Korea
Flavonoids
are
polyphenolic
compounds
presented
as
constituents
of
plants,
particularly
of
food
plants.
More
than
4,000
flavonoids
have
been
found
in
fruits,
vegetables,
and
beverages.
And
some
of
them
were
identified
as
significant
natural
antioxidants
[1].
In
the
bioactivity
studies,
structural
features
such
as
number
and
position
of
hydroxyl
groups
or
other
modifi-‐
cations
like
methylation,
prenylation,
glycosylation,
etc.,
are
key
factors
influencing
the
selec-‐
tivity
and
strength
of
potential
activities.
Therefore,
the
impact
on
the
biological
activities
of
flavonoids
can
hardly
be
predicted
with
even
minor
modifications
[2,
3].
In
the
present
study,
the
biotransformation
of
a
series
of
prenylated
flavonoids
was
investigated
with
twenty
dif-‐
ferent
microbial
strains
to
discover
new
metabolites.
It
was
revealed
that
Mucor
hiemalis
was
the
most
appropriate
microorganism
which
was
capable
of
transforming
these
flavonoids.
Structures
of
the
new
metabolites
were
elucidated
as
4’-O-prenylquercetin 3-O-β-D-
glucopyranoside and 7-O-prenylquercetin 3-O-β-D-glucopyranoside by
spectroscopic
methods.
The
microbial
metabolites
and
their
respective
parent
compounds
were
further
evaluated
for
their
antioxidant
capacity
by
DPPH
assay.
It
was
concluded,
however,
that
decreased
activity
of
the
metabolites
suggested
that
the
free
hydroxyl
group
at
C-‐3
position
was
a
crucial
factor
for
the
antioxidant
activity.
In addition, the existence of a flavonoid glucosidase was indicated for
M. hiemalis which may provide a potent approach to prepare glucosylated products of free flavo-
noids.
Acknowledgements:
Chonnam
National
University
Center
for
Research
Facilities
is
acknowledged
for
running
NMR
experiments
Keywords: Biotransformation, prenlyated flavonoids, antioxidative activity
References:
[1] Cao
G,
Sofic
E,
Prior
RL.
Antioxidant
and
prooxidant
behavior
of
flavonoids:
structure-‐
activity
relationships.
Free
Radic
Biol
Med
1997;
22:
749−760
[2] Heim
KE,
Tagliaferro
AR,
Bobilya
DJ.
Flavonoid
antioxidants:
chemistry,
metabolism
and
structure-‐activity
relationships.
J
Nutr
Biochem
2002;
13:
572−584
[3] Schlupper
D,
Giesa
S,
Gebhardt
R.
Influence
of
biotransformation
of
luteolin,
luteolin
7-‐O-‐
glucoside,
3',4'-‐dihydroxyflavone
and
apigenin
by
cultured
rat
hepatocytes
on
antioxida-‐
tive
capacity
and
inhibition
of
EGF
receptor
tyrosine
kinase
activity.
Planta
Med
2006;
72:
596−603
P423
From
traditional
uses
to
phytochemical
study
of
a
Polynesian
healing
plant:
Constituents
and
properties
of
“Metuapua’a”
Raimana
Ho1,
Taivini
Teai1,
Jean-‐Pierre
Girault2,
Alain
Meybeck3,
René
Lafont4,
Phila
Raharive-‐
lomanana1
1
Université
de
la
Polynésie
Française,
Tahiti,
98702
FAA'A,
Polynésie
Française,
2
Laboratoire
de
Chimie
et
Biochimie
Pharmacologiques
et
Toxicologiques,
CNRS
UMR
8601,
Université
Paris
Descartes,
45
rue
des
Saints
Pères,
75270
Paris,
Cedex
06,
France,
3
AM
Phyto-‐Conseil,
20
ter
rue
de
Bezons,
92400
Courbevoie,
France,
4
Laboratoire
BIOSIPE,
ER3,
Paris
6
–
Pierre
et
Marie
Curie,
Case
29,
7
Quai
Saint
Bernard,
75252
Paris,
Cedex
5,
France
“Metuapua’a”
is
the
vernacular
name
attributed
to
distinct
fern
species
(Microsorum
grossum)
(Polypodiaceae)
used
in
a
great
number
of
remedies
in
Polynesian
traditional
medicine.
Fronds
and/or
rhizomes
are
usually
prescribed
to
treat
stomach
ache,
gonorrhoea,
pneumo-‐
nia,
leucorrhoea,
sterility,
dislocations,
fractures,
etc
[1].
Phytochemical
studies
of
these
spe-‐
cies
led
to
establishing
their
chemical
composition,
showing
that
the
main
components
are
purportedly
adaptogenic
phytoecdysteroid
such
as
ecdysone
and
20-‐hydroxyecdysone.
Their
complex
phytoecdysteroid
composition
and
distribution
differs
between
species
and
plant
organs
[2-‐5].
The
list
of
biological
properties
of
the
main
phytoecdysteroid
constituents
whose
benefits
on
health
are
well
known
[6-‐8]
justify
their
medicinal
uses.
The
skin-‐active
effect
of
M.
grossum
extract
was
investigated
in
two
ways
on
human
dermal
fibroblasts:
a
transcriptomic
study
with
c-‐DNA
array
for
gene
expression
modulation
and
a
stress
induced
premature
senescence
(SIPS)
test.
The
total
extract
of
M.
grossum
up
regulates
Heme
Oxygen-‐
ase
1
(HO1),
an
enzyme
which
protects
cells
from
oxidative
stress
and
which
is
responsible
for
skin
photoimmunoprotection.
We
report
that
premature
senescence
of
human
skin
in-‐
duced
by
repeated
UV
irradiations
can
be
prevented
by
an
exdysteroid
fraction
of
M.
grossum.
It
seems
therefore
that
extracts
of
this
fern
could
protect
skin
against
oxidative
stresses
and
that
so
could
be
used
as
innovative
active
cosmetic
ingredient
[9].
Acknowledgements:
Financial
support
from
the
“Délégation
de
la
Recherche”
of
French
Polynesia
and
from
the
“Ministère
de
l’Outre-‐Mer”
(M.O.M.)
of
France
are
deeply
acknowledged.
Keywords: Microsorum grossum, ethnopharmacology, medicinal plant, phytoecdysteroids,
dermal fibroblast, cellular senescence
References:
[1] Pétard
P.
Plantes
utiles
de
Polynésie
et
Raau
Tahiti.
Ed.
Revue
et
augmentée,
Papeete,
Haere
Po
No
Tahiti
1986;
77-‐78
[2] Ho
R,
Teai
T,
Loquet
D,
Bianchini
JP,
Girault
JP,
Lafont
R,
Raharivelomanana
P.
Phytoecdys-‐
teroids
in
the
genus
Microsorum
(Polypodiaceae)
of
French
Polynesia,
Nat
Prod
Commun
2007;
2:
803-‐806
[3] Snogan
E,
Vahirua-‐Lechat
I,
Ho
R,
Bertho
G,
Girault
JP,
Ortiga
S,
Maria
A,
Lafont
R.
Ecdys-‐
teroids
from
the
medicinal
fern
Microsorum
scolopendria
(Burm.
F.),
Phytochem
Anal
2007;
18:
441-‐450
[4] Lafont
R,
Ho
R,
Raharivelomanana
P,
Dinan
L.
“Ecdysteroids
in
ferns:
distribution,
diversi-‐
ty,
biosynthesis
and
functions”
2010;
Chapter
22,
pp
305-‐319,
in
“Working
with
ferns,
is-‐
sues
and
applications”,
Fernandez
H,
Revilla
MA,
Kumar
A
(eds),
New
York
[5] Ho
R,
Girault
JP,
Raharivelomanana
P,
Lafont
R.
“E-‐
and
Z-‐isomers
of
new
phytoecdyster-‐
oid
conjugates
from
French
Polynesian
Microsorum
membranifolium
(Polypodiaceae)
fronds”.
Molecules
2012;
17:
11598-‐11606
[6] Lafont
R.,
Dinan
L.
Practical
uses
for
ecdysteroids
in
mammals
including
humans:
an
up-‐
date.
J
Insect
Science
2003;
3:
30
[7] Detmar
M.,
Dumas
M.,
Bonté
F.,
Meybeck
A.,
Orfanos
C.E.
Effects
of
ecdysterone
on
the
dif-‐
ferentiation
of
normal
human
keratinocytes
in
vitro.
Eur
J
Dermatol
1994;
4:
558-‐562
[8] Báthori
M,
Tóth
N,
Hunyadi
A,
Márki
A,
Zádor
E.
Phytoecdysteroids
and
anabolic-‐
androgenic
steroids-‐structure
and
effects
on
humans.
Curr
Med
Chem
2008;
15:
75-‐91
[9] Ho
R,
Teai
T,
Meybeck
A,
Raharivelomanana
P.
UV-‐protective
effects
of
phytoecdysteroids
from
Microsorum
grossum
extracts
on
human
dermal
fibroblasts.
Nat
Prod
Commun
2015;
10:
33-‐36
P424
Antiprotozoal
activity
against
Entamoeba
histolytica
of
furocou-‐
marins
isolated
from
Ruta
chalepensis
Ramiro
Quintanilla-‐Licea1,
Benito
D.
Mata-‐Cárdenas2(†),
Javier
Vargas-‐Villarreal3,
Aldo
F.
Ba-‐
zaldúa-‐Rodríguez1,
María
J.
Verde-‐Star1
1Facultad
de
Ciencias
Biológicas,
Universidad
Autónoma
de
Nuevo
León,
UANL,
Av.
Universidad
S/N,
Cd.
Universitaria,
San
Nicolás
de
los
Garza,
C.P.
66451
Nuevo
León,
Mexico,
2Facultad
de
Ciencias
Químicas,
Universidad
Autónoma
de
Nuevo
León,
UANL,
Av.
Universidad
S/N,
Cd.
Universitaria,
San
Nicolás
de
los
Garza,
C.P.
66451
Nuevo
León,
Mexico,
3Laboratorio
de
Bioquímica
y
Biología
Celular,
Centro
de
Investi-‐
gaciones
Biomédicas
del
Noreste
(CIBIN),
Dos
de
abril
esquina
con
San
Luis
Potosí,
C.P.
64720
Monterrey,
Mexico
Amoebiasis
caused
by
Entamoeba
histolytica,
a
protozoan
of
the
family
Endomoebidae
is
asso-‐
ciated
with
high
morbidity
and
mortality
and
is
therefore
considered
as
the
third
parasitosis
of
medical
importance
after
malaria
and
schistosomiasis
[1,
2].
Currently
metronidazole
is
the
therapeutic
drug
of
choice
for
the
treatment
of
amoebiasis,
but
is
experiencing
drug
re-‐
sistance
by
E.
histolytica,
resulting
in
the
need
for
increased
doses
to
overcome
the
infection
and
thus
causing
unpleasant
side
effects.
For
this
reason
new,
more
effective
and
safer
anti-‐
protozoal
agents
are
urgently
required
[3].
We
reported
shortly
the
antiamoebic
activity
in
vitro
of
the
methanolic
extract
of
Ruta
cha-‐
lepensis
(Rutaceae)
[4]
and
we
are
now
reporting
the
bioguided
isolation
of
compounds
with
amebicide
activity
from
this
plant.
Partition
of
the
methanolic
extract
of
Ruta
chalepensis
by
extraction
with
n-‐hexane
led
to
a
residue
with
good
activity
against
E.
histolytica
(93.0
%
growth
inhibition).
Chromatography
of
this
hexane
residue
over
a
silica
gel
column
led
to
the
isolation
of
chalepensin
(1),
whereas
the
following
work
up
of
the
methanol
residue
by
parti-‐
tion
between
methanol
and
ethyl
acetate
followed
by
chromatography
of
the
EtOAc
residue
(84.8
%
growth
inhibition)
over
a
silica
gel
column
afforded
chalepin
(2)
and
rutamarin
(3)
as
well
as
the
isoquinoline
alkaloid
graveoline
(4).
Identification
of
the
isolated
compounds
was
based
on
spectroscopic/spectrometric
analyses
and
comparison
with
literature
data.
These
compounds
were
tested
for
antiprotozoal
activity
against
E.
histolytica
tropohzoites
using
in
vitro
tests.
Vials
were
incubated
for
72
h.
The
inhibition
percentage
was
estimated
as
the
number
of
dead
cells
compared
with
the
untreated
controls.
IC50
of
these
compounds
was
determined
by
using
a
Probit
analysis
with
a
95
%
confidence
level.
The
isolated
furocoumarins
(Figure
1)
from
R.
chalepensis
displayed
more
than
90
%
growth
inhibition
against
E.
histolytica
at
a
concentration
of
150
µg/mL,
with
IC50
values
ranging
from
6.54
to
38.71
µg/mL,
far
less
effective
than
metronidazole
(IC50
0.205
µg/mL),
but
these
IC50
values
are
suitable
as
selection
criterion
for
further
investigation
of
this
plant
as
source
of
potential
antiprotozoal
agents.
Graveoline
did
not
show
amebicide
activity.
Chalepensin
(1)
Chalepin
(2)
Rutamarin
(3)
Graveoline
(4)
IC50
38.71
µg/mL
IC50
28.67
µg/mL
IC50
6.54
µg/mL
No
amebicide
activity
Figure
1.
Structure
and
IC50
against
E.
histolytica
of
compounds
isolated
from
Ruta
chalepensis
Acknowledgements:
The
authors
would
like
to
thank
the
DAAD
(Germany),
who
financed
a
stay
of
R.Q.L.
at
the
University
of
Göttingen,
and
to
CONACYT
(Mexico)
for
doctoral
fellowship
awarded
to
A.F.B.R.
Keywords: neglected diseases, amoebiasis, Mexican medicinal plants, antiprotozoal agents
References:
[1] Hotez
PJ,
Woc-‐Colburn
L,
Bottazzi
ME.
Neglected
tropical
diseases
in
Central
America
and
Panama:
Review
of
their
prevalence,
populations
at
risk
and
impact
on
regional
develop-‐
ment.
Int
J
Parasitol
2014;
44:
597–603
[2] Bansal
D,
Sehgal
R,
Chawla
Y,
Malla
N,
Mahajan
RC.
Multidrug
resistance
in
amoebiasis
patients.
Indian
J
Med
Res
2006;
124:
189–194
[3] Singh
S,
Bharti
N,
Mohapatra
PP.
Chemistry
and
biology
of
synthetic
and
naturally
occur-‐
ring
antiamoebic
agents.
Chem
Rev
2009;
109:
1900–1947
[4] Quintanilla-‐Licea
R,
Mata-‐Cárdenas
BD,
Vargas-‐Villarreal
J,
Bazaldúa-‐Rodríguez
AF,
Ánge-‐
les-‐Hernández
IK,
Garza-‐González
JN,
Hernández-‐García
ME.
Antiprotozoal
activity
against
Entamoeba
histolytica
of
plants
used
in
northeast
Mexican
traditional
medicine.
bioactive
compounds
from
Lippia
graveolens
and
Ruta
chalepensis.
Molecules
2014;
19:
21044-‐21065
P425
A
new
inhibitor
of
hepatitis
C
virus
replication
from
Juncus
mari-‐
timus,
a
Tunisian
extremophile
plant
Ramla
Sahli1,2,
Céline
Rivière1,
Marie-‐Emmanuelle
Sahuc3,
Smaoui
Abderrazak2,
Jennifer
Samaillie1,
Vincent
Roumy3,
Thierry
Hennebelle3,
Yves
Rouillé3,
Karin
Seron3,
Sevser
Sahpaz1*,
Riadh
Ksouri2*
1Univ.
Lille,
INRA,
ISA,
Univ.
Artois,
Univ.
Littoral
Côte
d’Opale,
EA
7394
-‐ICV
-‐
Institut
Charles
Viollette,
F-‐
59000
Lille,
France,
2The
Laboratory
of
Aromatic
and
Medicinal
Plants,
Biotechnology
Centre
of
Borj-‐
Cédria
(CBBC),
Hammam-‐lif,
Tunisia,
3Univ.
Lille,
CNRS,
Inserm,
CHU
Lille,
Institut
Pasteur
de
Lille,
U1019
–
UMR
8204
–
CIIL
-‐
Center
for
Infection
and
Immunity
of
Lille
F-‐59000
Lille,
France
*
Both
authors
contributed
equally
to
this
work.
Hepatitis
C
virus
(HCV)
infection
is
a
major
cause
of
chronic
liver
diseases.
A
new
generation
of
anti-‐HCV
drugs
(viral
protein
inhibitors)
with
a
superior
efficacy
and
a
better
safety
profile
is
currently
available.
However,
they
are
very
expensive
and
not
accessible
in
developing
countries.
Also,
the
emergence
of
resistant
HCV
variants
continues
to
be
a
public
health
issue
[1].
The
discovery
of
new
HCV
inhibitors
remains
necessary
to
lower
the
cost
of
treatment
and
deepen
the
understanding
of
HCV
life
cycle.
In
this
context,
several
crude
methanolic
extracts
of
extremophile
plants
from
Tunisia
were
screened
against
HCV
in
cell
culture.
Infection
and
replication
rates
in
Huh-‐7
hepatoma
cells
were
investigated
by
indirect
immunofluorescence
assay.
The
rhizome’s
crude
extract
of
Jun-‐
cus
maritimus
Lam.,
a
halophyte
plant
belonging
to
Juncaceae
family,
exhibited
the
highest
activity,
especially
against
HCV
replication
(relative
infection
<10%
at
25
µg/mL).
Bioguided
fractionation
showed
that
the
methylene
chloride
partition
was
most
likely
responsible
for
this
activity
(relative
infection
=
6.26%
at
25
µg/mL).
From
this
partition,
several
compounds
were
isolated
by
CPC
and
preparative
HPLC
and
then
tested
against
HCV.
One
of
these
com-‐
pounds,
dehydrojuncusol,
a
known
phenanthrene
derivative
[2],
identified
by
HR-‐MS
and
NMR,
was
the
most
active.
Compared
to
DMSO-‐treated
cells,
dehydrojuncusol
significantly
inhibited
HCV
infection
when
added
during
the
inoculation
step
(IC50
=
5.56
µM),
and
even
more
when
added
during
the
post-‐inoculation
(replication)
step
(IC50
=
1.31
µM).
Its
effect
against
HCV
replication
was
sim-‐
ilar
to
boceprevir
(0.5
µM),
an
inhibitor
of
the
viral
polymerase
used
as
positive
control.
Moreover,
it
showed
no
in
vitro
toxicity
on
Huh-‐7
cells
at
active
concentrations.
Few
natural
products
have
demonstrated
activity
on
viral
replication
of
HCV
[3].
Activity
assays
against
HCV
resistant
mutants
are
underway
to
help
us
understanding
the
dehydrojuncusol
mecha-‐
nism
of
action.
Acknowledgements:
The
authors
wish
to
thank
CUMA
(Pr.
J.F.
Goossens)
and
LARMN
(Pr.
N.
Azaroual)
from
University
of
Lille
2
(France)
for
access
to
equipment
and
the
members
of
these
platforms
for
their
skillfull
technical
assistant.
Keywords: phenanthrene, Juncus maritimus, antiviral activity, HCV, extremophile plants
References:
[1] Galani
BRT,
Sahuc
ME,
Sass
G,
Njayou
FN,
Loscher
C,
Mkounga
P,
Deloison
G,
Brodin
P,
Rouillé
Y,
Tiegs
G,
Seron
K,
Moundipa
PF.
Khaya
grandifoliola
C.DC:
a
potential
source
of
active
ingredients
against
hepatitis
C
virus
in
vitro.
Arch
Virol
2016;
161:
1169–81
[2] Sarkar
H,
Zerezchi
M,
Bhattacharyya
J.
Dehydrojuncusol,
a
constituent
of
the
roots
of
Jun-‐
cus
roemerianus.
Phytochemistry
1988;
27:
3006–3008
[3] Calland
N,
Dubuisson
J,
Rouillé
Y,
Séron
K.
Hepatitis
C
Virus
and
Natural
Compounds:
a
New
Antiviral
Approach?
Viruses
2012;
10:
2197–2217
P426
Chemical
composition,
anti-‐inflammatory
and
antioxidant
activi-‐
ty
activities
of
essential
oil
of
Piper
cubeba
L.
Ramzi
Mothana1,
Mansour
AlSaid1,
Mohammad
Raish2,
Mohammed
Al-‐Sohaibani3,
Ajaz
Ah-‐
mad4,
Mohammed
Al-‐Yahya1,
Jamal
Kaled5,
Syed
Rafatullah1
1
Department
of
Pharmacognosy
and
Medicinal,
Aromatic
&
Poisonous
Plants
Research
Center
(MAP-‐
PRC),
College
of
Pharmacy,
P.O.
Box
2457,
King
Saud
University,
Riyadh
11451,
Saudi
Arabia.
2
Depart-‐
ment
of
Pharmaceutics,
College
of
Pharmacy,
P.O.
Box
2457,
King
Saud
University,
Riyadh
11451,
Saudi
Arabia.
3
Department
of
Medicine
and
Pathology,
Gastroenterology
Unit,
Collage
of
Medicine,
King
Khalid
University
Hospital,
King
Saud
University
P.O.
Box
2925,
Riyadh-‐11461
Saudi
Arabia.
4
Department
of
Clinical
Pharmacy,
College
of
Pharmacy,
P.O.
Box
2457,
King
Saud
University,
Riyadh
11451,
Saudi
Ara-‐
bia.
5
Departments
of
Botany
and
Microbiology,
College
of
Science,
King
Saud
University,
Riyadh
11451,
Saudi
Arabia
Piper
cubeba
(L.)
is
popularly
used
as
herbal
remedy
for
various
ailments.
However,
the
scien-‐
tific
basis
for
its
medicinal
use
especially
in
inflammation
remains
unknown.
Therefore,
the
present
study
was
aimed
to
investigate
the
anti-‐inflammatory
and
antioxidant
activity
of
Piper
cubeba
essential
oil
(PCEO)
in
laboratory
rodent
models.
The
in
vivo
anti-‐inflamatory
activity
of
PCEO
at
three
doses
(150,
300
and
600
mg/kg,
p.o)
was
tested
in
carrageenan-‐induced
rat
paw
edema,
cotton
pellet
granuloma
and
carrageenan-‐induced
pleurisy.
The
mechanism
of
inflammation
using
carrageenan-‐induced
pleurisy
in
rat
was
further
studied.
The
in
vitro
anti-‐
oxidant
activity
was
examined
using
DPPH.
In
parallel
to
that,
the
oil
was
analyzed
by
GC/MS
investigation.
PCEO
at
600
mg/kg
reduced
the
paw
edema
considerably
(65%)
and
the
weight
of
cotton
pellet
granuloma
(46%).
The
results
indicated
that
the
pretreatment
with
PCEO
at
doses
of
150,
300
and
600
mg/kg
reduced
exudate
volume
to
1.15,
0.89
and
0.73
ml
respec-‐
tively.
The
number
of
polymorphonuclear
(PMN)
cells
was
also
reduced
to
52.9
×
106,
43.49
×
106
and
37.37
×
106,
respectively.
Furthermore,
dose
dependent
reduction
in
myeloperoxi-‐
dase
(MPO),
nitric
oxide
and
proinflammatory
cytokine
such
tumor
necrosis
factor
(TNFα)
and
interleukin-‐1
(IL-‐1β)
was
observed
and
supported
by
histological
observation.
Moreover,
PCEO
exhibited
promising
strong
antioxidant
as
demonstrated
in
DPPH
assay.
The
GC/MS
revealed
that
monoterpenes
e.g.
sabinene,
4-‐terpineol,
γ-‐terpinene
and
α-‐thujene
were
the
major
components
in
PCEO
which
could
be
responsible
for
the
observed
activities.
Acknowledgements:
The
authors
extend
their
appreciation
to
the
Deanship
of
Scientific
Research
at
King
Saud
University
for
funding
the
work
through
the
research
group
project
No.
(RGP-‐VPP-‐073).
Keywords:
Piper
cubeba,
essential
oil,
anti-‐inflammatory,
antioxidant,
proinflammatory
cy-‐
tokines.
P427
Identification
of
phenolic
and
volatile
compounds
in
Centhrant-‐
hus
longiflorus
subsp.
longiflorus
by
HPLC
and
GC-‐MS,
antioxidant
and
antimicrobial
activity
studies
Sıla
Özlem
Şener1,
Merve
Badem1,
Nuriye
Korkmaz2,
Rezzan
Aliyazicioglu2,
Ufuk
Özgen1,
Şen-‐
gül
Alpay
Karaoğlu3
1
Department
of
Pharmacognosy,
Karadeniz
Technical
University,
Faculty
of
Pharmacy,
61080
Trabzon,
Turkey,
2
Department
of
Biochemistry,
Karadeniz
Technical
University,
Faculty
of
Pharmacy,
61080
Trabzon,
Turkey,
3
Department
of
Biology,
Recep
Tayyip
Erdoğan
University,
Faculty
of
Science,
53100
Rize,
Turkey
The
genus
Centranthus
belongs
to
Valerianaceae
family.
Three
species
of
this
genus
are
found
in
Turkey:
C.
ruber,
C.
longiflorus
and
C.
calcitrapa.1
C.
longiflorus
subsp.
longiflorus
is
used
for
sleep
disorders
in
traditional
Turkish
medicine.2
The
aim
of
this
study
was
to
determine
the
total
phenolic
contents
by
using
RP-‐HPLC,
volatile
compounds
by
using
GC-‐MS
and
detect
an-‐
tioxidant
activity
and
antimicrobial
activity.
Octanal,
γ-‐terpinene,
limonene,
2E,
4Z-‐
decadienal,
α–cubebene,
E-‐caryophyllene,
γ–muurolene,
abietadiene,
docosane
and
san-‐
daracopimarinal
were
specified
by
GC-‐MS.
Phenolic
compounds
detected
were
protocatechuic
acid,
vanillin
and
rosmarinic
acid.
The
most
powerful
antimicrobial
activity
was
observed
on
Mycobacterium
smegmatis.
According
to
the
results,
aqueous
extract
exhibited
powerful
anti-‐
oxidant
activity
and
showed
moderate
antimicrobial
activity.
Acknowledgements:
The
authors
would
like
to
thank
TÜBİTAK
for
its
financial
support
Keywords:
Centhranthus,
RP-‐HPLC,
GC-‐MS,
phenolic
compounds,
antioxidant
and
antimicro-‐
bial
activity.
References:
[1] Richardson
IBK.
Centhranthus
DC.
In
flora
of
Turkey
and
East
Aegean
Islands,
Ed.
Davis
PH.
Edinburgh
University
Press,
Edinburgh,
UK
1972;
4:
558
[2] Baytop
T.
Theraphy
with
Medicinal
Plants
in
Turkey,
Nobel
Tıp
Basımevi,
Istanbul,
Tur-‐
key,
1999
P428
Phenolic
composition
by
RP-‐HPLC
and
antioxidant
capacity
stu-‐
dies
of
Consolida
orientalis
(Gay)
Schrod.
Rezzan
Aliyazicioglu1,
Ufuk
Özgen2,
Merve
Badem2,
Sıla
Özlem
Şener2,
Nuriye
Korkmaz1,
Şengül
Alpay
Karaoğlu3
1
Department
of
Biochemistry,
Karadeniz
Technical
University,
Faculty
of
Pharmacy,
61080
Trabzon,
TURKEY,
2
Department
of
Pharmacognosy,
Karadeniz
Technical
University,
Faculty
of
Pharmacy,
61080
Trabzon,
TURKEY,
3
Department
of
Biology,
Recep
Tayyip
Erdoğan
University,
Faculty
of
Science,
53100
Rize,
TURKEY
The
genus
Consolida
belongs
to
Ranunculaceae
and
includes
about
52
species
world-‐wide[1].
C.
orientalis,
a
member
of
the
genus,
was
investigated
in
terms
of
phenolic
contents
and
anti-‐
oxidant
capacity
in
the
present
study.
RP-‐HPLC
was
used
for
the
identification
of
the
phenolic
compounds
of
C.
orientalis.
Antioxidant
capacity
were
determined
by
using
three
different
methods
test
of
the
plant,
including
FRAP
assay
(Ferric
reducing
antioxidant
potential),
DPPH
radical
scavenging
assay
(1,1-‐diphenyl-‐2-‐picryl
hydrazyl
radical
reducing
power
methods),
and
total
phenolic
contents.
Phenolic
compounds
were
detected
p-‐hydroxy
benzoic
acid,
chlorogenic
acid,
caffeic
acid,
p-‐coumaric
acid
and
sinapic
acid.
The
strongest
antimicrobial
activities
were
observed
on
Yersinia
pseudotuberculosis
and
Mycobacterium
smegmatis.
Our
results
showed
that
the
plant
may
be
used
as
a
protective
agent
against
diseases
as
well
as
in
food
industries.
Acknowledgements:
The
authors
would
like
to
thank
TÜBİTAK
for
its
financial
support
Keywords:
RP-‐HPLC,
Phenolic
compounds,
Antioxidant
capacity,
Antimicrobial
activity
References:
[1] Tavassoli
A,
Pakravan
M,
Kiarostami
K,
Poorhabibian
R.
Karyotype
analysis
in
some
spe-‐
cies
of
Consolida
(Ranunculaceae)
from
Iran.
Feddes
Repertorium
2013,
123:
257-‐263
P429
HPLC
Profiles
of
phenolic
compounds,
antioxidant
and
antimic-‐
robial
activity
of
the
aerial
parts
of
Coronilla
varia
L.
Rezzan
Aliyazicioglu1,
Ufuk
Özgen2,
Sıla
Özlem
Şener2,
Merve
Badem2,
Nuriye
Korkmaz1,
Şengül
Alpay
Karaoğlu3
1
Department
of
Biochemistry,
Karadeniz
Technical
University,
Faculty
of
Pharmacy,
61080
Trabzon,
Turkey,
2
Department
of
Pharmacognosy,
Karadeniz
Technical
University,
Faculty
of
Pharmacy,
61080
Trabzon,
Turkey,
3
Department
of
Biology,
Recep
Tayyip
Erdoğan
University,
Faculty
of
Science,
53100
Rize,
Turkey
Coronilla
genus
has
about
50
species
primarily
centred
around
the
Mediterranean,
extending
to
the
Atlantic
Islands,
Western
Asia
and
Northeast
Africa
[1].
It
has
seven
species
in
Turkey
[2].
Reverse
phase-‐high
performance
liquid
chromatography
(RP-‐HPLC)
was
used
for
deter-‐
mination
of
phenolic
compounds
in
the
methanolic
extract
of
aerial
parts
of
C.
varia.
The
anti-‐
oxidant
capacity
were
assessed
by
the
2,2-‐diphenyl-‐1-‐picrylhydrazyl
(DPPH),
and
ferric
re-‐
ducing
antioxidant
power
(FRAP)
tests.
Total
phenol
content
of
methanolic
extract
of
the
plant
was
calculated
spectrophotometrically.
The
antimicrobial
activity
was
determined
using
the
disc
diffusion
method.
The
IC50
value
for
DPPH
assay
has
been
found
as
0,708 ± 0,010
(mg/mL),
FRAP
value
is
240 ± 2,645 (μM
Trolox/g
sample),
and
total
phenolic
content
value
is
11,2 ± 0,472 mg
gallic
acid
per
gram
sample
in
methanolic
extract
of
the
aerial
parts
of
the
C.
varia.
Phenolic
compounds
were
identified
as
chlorogenic
acid,
vanillic
acid,
caffeic
acid,
vanil-‐
lin,
syringaldehyde,
p-‐coumaric
acid,
ferulic
acid,
sinapic
acid,
and
benzoic
acid.
The
meth-‐
anolic
extract
exhibited
moderate
antibacterial
activity
against
an
acid-‐fast
bacterium
(M.
smegmatis),
and
Gram
negative
(Y.
pseudotuberculosis)
bacteria.
Antifungal
activity
was
not
determined
against
C.
albicans
and
S.
cerevisiae.
According
to
the
results,
the
methanolic
ex-‐
tract
of
the
aerial
parts
of
the
plant
showed
potent
antioxidant
activity
and
moderate
antimi-‐
crobial
activity.
Keywords:
Coronilla
varia,
RP-‐HPLC,
antioxidant
capacity,
antimicrobial
activity
References:
[1] Diez
MJ,
Ferguson
IK.
Studies
of
the
pollen
morphology
and
taxonomy
of
the
tribes
Loteae
and
Coronilleae
(Papilionoideae;
Leguminoseae),
Coronilla
L.
and
related
genera
and
sys-‐
tematic
conclusions.
Rev
Palaeobot
Palyno1996;
94:
239-‐257.
[2] Chamberlain
DF.
Coronilla
L.
Flora
of
Turkey
and
the
East
Aegean
Islands.
Ed.
Davis
PH,
University
Press,
Edinburg
1970
3:
538.
P430
Antimicrobial
and
antioxidant
activities
of
the
aerial
parts
and
roots
of
Cirsium
trachylepis
Rezzan
Aliyazicioglu1,
Ufuk
Özgen2,
Sıla
Özlem
Şener2,
Merve
Badem2,
Şengül
Alpay
Karaoğlu3
1
Department
of
Biochemistry,
Karadeniz
Technical
University,
Faculty
of
Pharmacy,
61080
Trabzon,
TURKEY,
2
Department
of
Pharmacognosy,
Karadeniz
Technical
University,
Faculty
of
Pharmacy,
61080
Trabzon,
TURKEY,
3
Department
of
Biology,
Recep
Tayyip
Erdoğan
University,
Faculty
of
Science,
53100
Rize,
TURKEY
Cirsium
Mill.
is
one
of
the
largest
members
of
the
family
Asteraceae
and
consists
of
approxi-‐
mately
250
species,
found
in
Europe,
North
Africa,
East,
Central,
and
Southwest
Asia
and
North
and
Central
America.
These
grow
in
a
wide
range
of
habitats,
from
coastal
regions
to
the
alpine
belt
[1,2].
Cirsium
genus
in
Turkey
is
represented
by
66
species
(78
taxa)
and
thirty
of
which
are
endemic.
C.
trachylepis
Boiss.
is
an
endemic
species
for
Turkey
[3].
The
aim
of
this
study
was
to
detect
antioxidant
activity
and
antimicrobial
activity
of
the
aerial
parts
and
roots
of
the
plant.
Antioxidant
activity
studies
(Ferric
Reducing
Antioxidant
Power
(FRAP)
and
radical
scavenging
activity
tests
DPPH)
and
antimicrobial
activity
study
(agar
well
diffu-‐
sion
assay)
were
performed
on
the
methanolic
extracts
of
the
aerial
parts
and
roots
of
the
plant.
The
IC50
values
for
DPPH
assay
have
been
found
as
0,151
±
0.018
and
0.070
±
0.014
mg/mL;
FRAP
values
are
73,8
±
2,7
and
71,5
±
6.2
μM
Trolox/g
sample
for
the
aerial
parts
and
roots
of
the
plant,
respectively.
The
methanolic
extracts
of
the
aerial
parts
and
the
roots
of
the
plant
showed
moderate
activity
on
E.
coli
ATCC
35218,
Y.
pseudotuberculosis
ATCC
911,
P.
ae-‐
ruginosa
ATCC
10145
and
M.
smegmatis
ATCC607.
Cirsium
trachylepis
Acknowledgements:
The
authors
would
like
to
thank
TÜBİTAK
(215Z026)
for
its
financial
support
Keywords:
Cirsium
trachylepis,
Asteraceae,
Antioxidant
activity,
Antimicrobial
activity
References:
[1] Kadereit
JW,
Jeffrey
C.
Flowering
Plants.
Eudicots:
Asterales.
In:
Kubitzki
K
(ed.)
“The
Families
and
Genera
of
Vascular
Plants”,
2007;
8:
132.
[2] Petrak
F.
Editor:
Rechinger,
K.
H.
Cirsium
Mill.
In
Flora
Iranica.
Compositae
III-‐Cynareae,
1979;
139:
231-‐280.
[3] Davis
PH,
Parris
BS.
Flora
of
Turkey
and
the
East
Aegean
Islands.
Editor:
Davis,
P.H.
Edin-‐
burgh:
Edinburgh
University
Press.
1975;
5:
382-‐383
P431
Evaluation
and
isolation
of
antimutagenic
compounds
from
methanolic
leaf
extracts
of
Monanthotaxis
caffra
Rhulani
Makhuvele1,2,
Kenn
Foubert3,
Sandra
Apers3,
Luc
Pieters3,
Luc
Verschaeve4,5,
Esameldin
E.
Elgorashi1,2
1
Toxicology
and
Ethnoveterinary
Medicine,
Agricultural
Research
Council-‐Onderstepoort
Veterinary
in-‐
stitute,
Private
Bag
X05,
Onderstepoort,
0110
Pretoria,
South
Africa,
2
Department
of
Paraclinical
Scienc-‐
es,
University
of
Pretoria,
Private
Bag
X04,
Onderstepoort,
0110
Pretoria,
South
Africa,
3
Department
of
Pharmaceutical
Sciences,
University
of
Antwerp,
Universiteitsplein
1,
2610
Antwerp,
Belgium,
4
Toxicolo-‐
gy,
Scientific
Institute
of
Public
Health,
Rue
Juliet
Wytsmanstraat
14,
1050
Brussels,
Belgium,
5
Depart-‐
ment
of
Biomedical
Sciences,
University
of
Antwerp,
2610
Antwerp,
Belgium
Plants
are
known
to
synthesize
therapeutic
compounds
which
can
modulate
or
prevent
muta-‐
genic
effects
[1].
Annonaceae
plant
species
have
been
used
in
traditional
medicine
to
treat
microbial
and
parasitic
[2]
infections
and
tumours
[3].
Monanthotaxis
caffra
(Sond.)
Verdc.
is
a
member
of
the
Annonaceae
family
reported
to
possess
antitumoural
properties
[4].
The
aim
of
this
study
was
to
evaluate
and
isolate
the
antimutagenic
compounds
from
the
methanolic
leaf
extract
of
M.
caffra.
The
leaves
of
M.
caffra
were
ground
to
fine
powder
and
extracted
with
90%
methanol.
The
crude
extract
was
then
investigated
in
vitro
for
antimutagenic
activity
against
the
mutagen
aflatoxin
B1
using
the
Vitotox,
Ames
and
Comet
assays
in
the
absence
and
presence
of
S9
rat
liver
fraction.
The
results
for
the
Ames
and
Vitotox
assays
showed
that
M.
caffra
possessed
a
strong
antimutagenic
effect
(≥
58%)
against
aflatoxin
B1
at
a
concentra-‐
tion
of
5
mg/ml
while
in
the
Comet
test,
no
DNA
damage
was
observed
at
all
tested
concentra-‐
tions.
These
results
strongly
reveal
the
antimutagenic
potential
of
M.
caffra.
Therefore,
the
crude
methanolic
leaf
extract
of
M.
caffra
was
subjected
to bioassay-guided fractionation, during
which liquid-‐liquid
partition,
thin
layer
chromatography
(TLC),
high
pressure
liquid
chroma-‐
tography
(HPLC),
flash
chromatography
and
preparative
high
pressure
liquid
chromatog-‐
raphy-‐mass
spectrometry
(HPLC-‐MS)
were
used
to
isolate
its
constituents.
The
isolated
com-‐
pounds
were
identified
using
nuclear
magnetic
resonance
(NMR)
spectroscopy
and
MS.
They
were
identified
as
crotepoxide
and
5,6-‐diacetoxy1-‐benzoyloxymethyl-‐1,3-‐cyclohexadiene.
Crotepoxide
has
been
reported
to
be
present
in
the
genus
Monanthotaxis
and
to
exhibit
tu-‐
mour
inhibitory
activity
[4].
Acknowledgements: Flemish Interuniversity Council -‐ University Development Cooperation (VLIR-‐UOS),
Fonds voor Wetenschappelijk Onderzoek -‐ Flanders (Belgium) (FWO), National Research Foundation
(NRF) and Agricultural Research Council for their financial support
Keywords: Annonaceae, Monanthotaxis caffra (Sond.) Verdc., antimutagenic activity, cro-‐
tepoxide, 5,6-‐diacetoxy1-‐benzoyloxymethyl-‐1,3-‐cyclohexadiene
References:
[1] Celik
TA.
Potential
genotoxic
and
cytotoxic
effects
of
plant
extracts,
a
compendium
of
es-‐
says
on
alternative
therapy.
Bhattacharya,
A
(Ed.),
InTech,
Turkey,
2012;
Chapter
11,
pp.
233-‐235.
[2] Parmena
DS,
Mgina
CA,
Joseph
CC.
Composition
of
non-‐volatile
oils
and
antimicrobial
ac-‐
tivities
of
extracts
from
Monanthotaxis
discolor,
and
an
undescribed
Uvariondendron
spe-‐
cies.
Tanz
J
Sci
2012;
38:
221-‐231
[3] Biba
VS,
Amily
A,
Sangeetha
S,
Remani
P.
Anticancer,
antioxidant
and
antimicrobial
activity
of
Annonaceae
family.
World
J
Pharmacy
Pharmaceut
Sci
2014,
3:1595-‐1604
[4] Mulholland
D,
Naidoo
N,
Hutchings
A,
Lavaud
C,
Massiot
G.
Crotepoxide,
a
cyclohexane
di-‐
epoxide
from
Monanthotaxis
caffra.
Biochem
Syst
Ecol
2000;
28:
595-‐597
P432
Potential
role
of
arbuscular
mycorrhizal
fungi
in
the
accumula-‐
tion
of
polyphenols
in
Lamiaceae
species
Rita
Engel1,
Krisztina
Szabó1,
László
Abrankó2,
Anna
Füzy3,
Tünde
Takács3
1
MTA,
Centre
for
Ecological
Research,
Institute
of
Ecology
and
Botany,
Alkotmány
út
2-‐4,
H-‐2163,
Vácrátót,
Hungary;
2
Department
of
Applied
Chemistry,
Faculty
of
Food
Science,
Corvinus
University
of
Budapest,
Villányi
út
29–43,
H-‐1118,
Budapest,
Hungary;
3
MTA
Centre
for
Agricultural
Research,
Insti-‐
tute
for
Soil
Sciences
and
Agricultural
Chemistry,
Herman
Ottó
út
15,
H-‐1022,
Budapest,
Hungary
The
aim
of
this
study
was
to
examine
the
effect
of
arbuscular
mycorrhizal
(AM)
fungi
coloni-‐
zation
on
the
biomass
and
polyphenol
content
of
two
economically
important
Lamiaceae
herbs.
In
this
regard
a
15-‐week
pot
experiment
was
performed
under
controlled
conditions
with
Origanum
majorana
L.
and
Melissa
officinalis
L.
applying
a
commercially
available
AM
fungi
mixture
(Glomus
etunicatum,
Glomus
claroideum,
Rhizophagus
intraradices)
for
inocula-‐
tion.
The
efficiency
of
mycorrhization
was
determined
[1,2].
The
major
polyphenols
of
the
two
plants
were
identified
and
quantified
using
HPLC-‐ESI-‐qTOFMS
and
HPLC-‐PDA
systems
[3].
From
the
aerial
parts
of
the
plants
the
major
polyphenols
were
tentatively
identified,
five
from
marjoram
(apigenin-‐6,8-‐di-‐C-‐glucoside,
luteolin-‐7’-‐O-‐glucuronide,
rosmarinic
acid,
apig-‐
enin-‐glucuronide,
lithospermic
acid
A
isomer,)
and
three
from
lemon
balm
(rosmarinic
acid
and
two
lithospermic
acid
A
isomers).
According
to
our
results
marjoram
had
higher
level
of
fungal
colonization
compared
to
lemon
balm.
AM
fungal
inoculation
significantly
increased
the
biomass
of
marjoram,
but
did
not
cause
differences
in
lemon
balm
yield.
In
the
term
of
polyphenol
content
the
mycorrhization
mostly
affected
the
accumulation
of
lithospermic
acid
A
isomers
and
rosmarinic
acid.
The
AM
fungal
colonization
significantly
increased
the
content
of
these
compounds
in
lemon
balm,
but
decreased
in
marjoram.
Take
in
account
the
yield
of
biomass
the
total
yield
of
polyphenols
was
increased
as
the
result
of
mycorrhization
in
both
plants.
The
results
of
the
present
study
show
that
AMF
can
provide
different
services
for
each
herb.
For
instance
while
marjoram
benefits
more
from
the
AMF
colonization
in
terms
of
growth,
lemon
balm
has
higher
content
of
phenolic
acids.
It
shows
that
the
effect
and
outcome
of
mycorrhization
depends
on
the
plant
and
also
the
fungal
species.
Thus
the
application
of
optimized
AM
fungal
inoculum
could
improve
the
quantity
and
quality
of
medicinal
plant
ma-‐
terial.
Acknowledgements:
Authors
thank
OTKA
(PD105750)
for
the
financial
support.
Keywords: Origanum majorana, Melissa officinalis, arbuscular mycorrhizal fungi, polyphenol
References:
[1] Phillips
JM,
Hayman
DS.
Improved
procedures
for
clearing
roots
and
staining
parasitic
and
vesicular-‐arbuscular
mycorrhizal
fungi
for
rapid
assessment
of
infection.
Trans
Br
Mycol
Soc
1970;
55:
158-‐161
[2] Trouvelot
A,
Kought
JI.
Mesure
du
taux
de
mycorhization
VA
d’un
système
radiculaire.
Recherche
de
méthodes
d’estimation
ayant
une
signification
fonctionnelle.
In:
Gianinazzi-‐
Pearson
V,
Gianinazzi
S,
editors.
Physiological
and
genetical
aspects
of
mycorrhizae,
INRA,
1986:
217-‐221
[3] Abranko
L,
Garcia-‐Reyes
JF,
Molina-‐Diaz
A.
Systematic
bottom-‐up
approach
for
flavonoid
derivative
screening
in
plant
material
using
liquid
chromatography
high-‐resolution
mass
spectrometry.
Anal
Bioanal
Chem
2012;
403:
995-‐1006
P433
Chemical
composition
and
bioactivity
of
the
essential
oil
of
Pinus
roxburghii
bark
Rola
M.
Labib1,2
,
Fadia
S.
Youssef1,
Mohamed
L.
Ashour1,
Jennifer
Búfalo3,4,
Samir
A.
Ross2,5
1Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Ain-‐Shams
University,
11566,
Cairo,
Egypt,
2National
Center
for
Natural
Products
Research,
University
of
Mississippi,
University,
Mississippi,
38677,
US,
3Institute
of
Biosciences
(IB),
UNESP-‐Univ.,
Estadual,
Paulista,
Botucatu,
Sao
Paulo,
Brazil,
4United
States
Department
of
Agriculture,
Agricultural
research
Service,
Natural
Products
Utilization
Research
Unit,
University
of
Mississippi,
University,
MS,
38677,
USA,
5Department
of
BioMolecular
Sciences,
School
of
Pharmacy,
University
of
Mississippi,
University,
MS,
38677,
USA
Essential
oils
are
volatile
secondary
metabolites
that
are
normally
produced
and
can
be
used
safely
to
treat
several
health
disorders
[1].
The
essential
oil
obtained
from
Pinus
roxburghii
Sarg.
bark
(Pinaceae)
cultivated
in
Egypt
was
qualitatively
and
quantitatively
analyzed
using
GC/FID
and
GC/MS
techniques
[2].
Seventy
five
components
representing
89.65%
of
the
total
hydrodistilled
oil
were
identified,
with
longifolene
(33.13%)
representing
the
major
compo-‐
nent.
The
sequiterpene
hydrocarbons
and
the
oxygenated
sequiterpenes
constitute
37.13
and
19.79%
respectively.
The
essential
oil
was
assessed
for
its
antimicrobial
activity
against
two
Gram
positive,
two
Gram
negative,
methicillin
resistant
Staphylococcus
aureus
(MRSA)
and
five
fungal
strains.
It
exhibited
mild
antimicrobial
activity
showing
IC50
values
exceeding
20
μg/mL.
Moreover,
in
vitro
antimalarial
activity
was
investigated
against
both
chloroquine
sen-‐
sitive
(D6)
and
resistant
(W2)
strains
of
Plasmodium
falciparum
in
which
the
oil
exerted
a
mild
activity.
The
antileishmanial
activity
was
evaluated
in
vitro
against
different
cultures
of
Leishmania
donovani
promastigotes
and
amastigotes.
It
was
also
tested
against
Trypanosoma
brucei,
the
oil
showed
a
mild
antileishmanial
and
trypanocidal
activities,
with
IC50
values
ex-‐
ceeding
10
μg/mL.
Concerning
its
anti-‐inflammatory
activity,
no
inhibitory
effect
of
the
essen-‐
tial
oil
on
the
cannabinoids
or
opioids
receptors
was
shown.
In
silico
molecular
docking
of
its
major
components
was
performed
on
human
glucocorticoids
receptor
(GR).
Results
clarified
that
longifolene,
possess
an
affinity
to
GR
binding
site
comparable
to
the
potent
anti-‐
inflammatory
GR
agonist,
dexamethasone.
The
binding
energy
showed
that
longifolene
has
the
higher
binding
affinity
(-‐28.76
kcal/mol)
revealing
a
significant
anti-‐inflammatory
activity
as
compared
to
the
other
oil
components.
Pinus
roxburghii
Sarg
Longifolene
Molecular
docking
of
longifolene
Acknowledgements:
We
are
thankful
to
Dr.
Babu
Tekwani,
Dr.
Shabana
Khan
and
Dr.
Melissa
Jacob,
Na-‐
tional
Center
for
Natural
Products
Research,
The
University
of
Mississippi,
School
of
Pharmacy,
Mississip-‐
pi,
USA
for
the
antileishmanial,
antimalarial
and
antimicrobial
assays.
Keywords: Pinus roxburghii, sssential oil, GC/MS, anti-‐infective, anti-‐inflammatory, molecular
docking.
References:
[1] Husnu
Can
Baser
K,
Buchbauer
G.
Handbook
of
essential
oils,
science,
technology
and
ap-‐
plications.
CRC
press,
2010
[2] Adams
RP.
Identification
of
essential
oil
components
by
gas
chromatography/
quadrupole
mass
spectroscopy.
Carol
Stream,
IL:
Allured
Pub
Corp,
2004
[3] El-‐Readi
MZ,
Eid
HH,
Ashour
ML,
Eid
SY,
Labib
RM,
Sporer
F,
Wink
M.
Variations
of
the
Chemical
Composition
and
Bioactivity
of
Essential
Oils
from
Leaves
and
Stems
of
Liquid-‐
ambar
styraciflua
(Altingiaceae).
J
Pharm
Pharmacol
2013;
65:
1653-‐1663
P434
Investigations
in
flavonoid
content
of
Camelina
sativa
(L.)
Crantz
during
seed
development
Roland
Molinié1,
Anthony
Quéro1,
David
Mathiron2,
Benjamin
Thiombiano1,
Jean-‐Xavier
Fon-‐
taine1,
Déborah
Brancourt1,
Olivier
Van
Wuytswinkel1,
Emmanuel
Petit1,
Hervé
Demailly3,
Gaëlle
Mongelard3,
Serge
Pilard2,
Brigitte
Thomasset4,
François
Mesnard1
1
Université
de
Picardie
Jules
Verne,
EA
3900-‐BIOPI
Biologie
des
Plantes
et
Innovation,
IUT
d’Amiens,
Département
Génie
Biologique,
Avenue
des
Facultés,
Le
Bailly
et
Faculté
de
Pharmacie,
1,
rue
des
Louvels,
80037
Amiens
cedex,
France,
2
Université
de
Picardie
Jules
Verne,
Plate-‐Forme
Analytique,
33
rue
Saint-‐
Leu,
80039
Amiens,
France,
3
Centre
de
Ressources
Régionales
en
Biologie
Moléculaire,
Université
de
Pi-‐
cardie
Jules
Verne,
80039
Amiens,
France,
4
CNRS-‐FRE
3580,
GEC,
Université
de
Technologie
de
Com-‐
piègne,
CS
60319,
60203
Compiègne
cedex,
France
In
recent
years,
Camelina
sativa
(L.)
Crantz
has
known
a
renewed
interest
for
its
agronomic
potential.
This
oilseed
crop
has
a
short
development
cycle
(85-‐100
days)
and
is
naturally
tol-‐
erant
to
biotic
and
abiotic
stresses
[1].
This
Brassicaceae
is
undemanding
in
fertilizer
input
and
requires
less
pesticide
compared
to
traditional
oilseed
crops
[2].
Currently
this
plant
is
mainly
grown
for
its
seeds
with
unusual
fatty
acid
profile
[3]
and
phytochemical
investiga-‐
tions
of
the
seeds
are
still
emerging.
In
order
to
complete
the
phytochemical
characterization
of
Camelina,
the
flavonoid
contents
of
seeds
has
been
extracted
by
the
successive
contribution
of
3
solvents
(MeOH,
CHCl3,
H2O).
The
hydroalcoholic
fraction
was
purified
by
HPLC
and
col-‐
lected
products
were
analyzed
by
LC-‐MS
and
NMR
to
be
identified.
This
method
allowed
the
characterization
of
two
novel
flavonoids
in
seeds
of
Camelina:
quercetin-‐5b-‐O-‐sinapyl-‐2"-‐O-‐
apiosyl-‐3-‐O-‐rutinoside
and
epicatechin-‐7-‐O-‐glucose.
These
2
compounds
have
a
contrasted
dynamic
accumulation
during
seed
development.
The
quercetin-‐5b-‐O-‐sinapyl-‐2"-‐O-‐apiosyl-‐3-‐O-‐rutinoside
accumulates
essentially
between
15
and
25
days
after
flowering
(DAF).
Thus,
the
area
ratio
was
0.1,
0.4
and
0.5
at
15,
25
and
35
DAF,
respectively.
In
contrast,
the
epicatechin-‐7-‐O-‐glucose
accumulates
essentially
between
25
and
35
DAF.
This
compound
was
not
detectable
at
15
and
25
DAF
and
the
area
ratio
was
3.7
at
35
DAF.
To
conclude,
this
study
allowed
to
characterize
two
novel
flavonoids
in
seeds
of
Camelina
and
to
describe
their
accumulation
during
seed
development.
Keywords:
Camelina
sativa,
seed,
quercetin-‐5b-‐O-‐sinapyl-‐2"-‐O-‐apiosyl-‐3-‐O-‐rutinoside,
epi-‐
catechin-‐7-‐O-‐glucose,
LC-‐MS,
NMR
References:
[1] Gugel
RK,
Falk
KC.
Agronomic
and
seed
quality
evaluation
of
Camelina
sativa
in
western
Canada.
Can
J
Plant
Sci
2006;
86:
1047–1058
[2] Moser
BR.
Camelina
(Camelina
sativa
L.)
oil
as
a
biofuels
feedstock:
Golden
opportunity
or
false
hope?
Lipid
Technol
2010;
22:
270–273
[3] Vollmann
J,
Moritz
T,
Kargl
C,
Baumgartner
S,
Wagentristl
H.
Agronomic
evaluation
of
camelina
genotypes
selected
for
seed
quality
characteristics.
Ind
Crops
Prod
2007;
26:
270–277
P435
Greek
flora
as
a
source
of
new
anti-‐oxidant,
anti-‐elastase,
anti-‐
collagenase
and
anti-‐hyaluronidase
natural
agents
Michalea
Rozalia,
Boka
Vasiliki-‐Ioanna,
Dina
Evanthia,
Aligiannis
Nektarios,
Skaltsounis
Alex-‐
ios-‐Leandros
Department
of
Pharmacognosy
and
Natural
Products
Chemistry,
Faculty
of
Pharmacy,
University
of
Ath-‐
ens,
Panepistimiopolis,
Zografou,
15771,
Athens,
Greece
In
order
to
investigate
new
cosmetic
ingredients
of
natural
origin,
50
plant
extracts
obtained
from
Greek
flora
were
screened
for
their
capacity
to
scavenge
free
radicals
and
inhibit
en-‐
zymes
related
to
skin
ageing.
Degradation
of
extracellular
matrix
(ECM)
has
directly
been
linked
to
skin
ageing
and
is
correlated
to
increase
in
activity
of
enzymes
such
as
elastase,
col-‐
lagenase
and
hyaluronidase.
Elastin,
collagen
and
hyaluronic
acid
decrease,
leading
to
a
loss
of
strength
and
flexibility
in
skin
and
formation
of
deep
wrinkles.
Also
free
radicals
induce
the
activity
of
these
enzymes
[1,2]. Discovery
of
bioactive
molecules
which
show
both
anti-‐
oxidant
and
anti-‐ageing
properties
is
the
target
of
the
present
study.
Anti-‐elastase,
anti-‐
collagenase
and
anti-‐hyaluronidase
activities
of
50
methanolic
and
ethyl
acetate
extracts
were
determined
using
spectrophotometric
methods.
Radical
scavenging
activity
was
determined
by
the
ability
of
the
extracts
to
scavenge
DPPH
and
ABTS
radicals.
The
majority
of
the
samples
especially
the
methanolic
extracts
showed
high
anti-‐oxidant
capacity,
e.g.
Rosa
damascena
extract
exhibited
IC50
value
at
47.6
μg/ml.
In
anti-‐elastase
assay
8
extracts
showed
more
than
50%
inhibition
at
100μg/ml,
in
anti-‐collagenase
assay
15
extracts
showed
more
than
60%
inhibiton
at
100μg/ml
and
in
anti-‐hyaluronidase
assay
8
extracts
showed
more
than
80%
in-‐
hibition
at
300μg/ml.
The
methanolic
extracts
of
Sedum
sediforme
and
Umbilicus
horizontalis
aerial
parts
inhibited
elastase
and
hyaluronidase
by
more
than
85%.
Rosa
damascena
inhibit-‐
ed
80%
of
collagenase
activity
at
100μg/ml.
Antioxidant
and
enzyme
inhibitory
activities
of
the
plant
extracts
suggest
that
they
can
restore
skin
elasticity
and
delay
wrinkling
process.
Bioguided
isolation
process
was
performed
in
order
to
reveal
the
active
secondary
metabo-‐
lites.
In
the
case
of
Rosa
damascena
quercetin-‐3-‐O-‐rhamnoside,
kaempferol-‐3-‐
glucopyranoside,
phenylethanol
were
proved
to
exhibit
high
anti-‐collagenase
activity.
Keywords:
Anti-‐ageing,
anti-‐collagenase,
anti-‐elastase,
anti-‐hyaluronidase,
anti-‐oxidant,
Greek
flora
References:
[1] Ndlovu
G,
Fouche
G.,
Tselanyane
M.,
Cordier
W.,
Steenkamp
V.
In
vitro
determination
of
the
anti-‐aging
potential
of
four
southern
African
medicinal
plants.
BMC
Complement
Altern
Med
2013;
13:
304
[2] Royer
M,
Prado
M,
Garcıa-‐Perez
ME,
Diouf
N,
Stevanovic
T.
Study
of
nutraceutical,
nutri-‐
cosmetics
and
cosmeceutical
potentials
of
polyphenolic
bark
extracts
from
Canadian
for-‐
est
species.
PharmaNutrition
2013;
1:
158-‐167
P436
Search for leishmanicidal agents by bioactivity-‐correlated tech-‐
niques from terrestrial plants
Manisha Bhatnagar1, Nandan Sarkar1, Sabari Ghosal1
1 Center for Plant and Environmental Biotechnology, Amity Institute of Biotechnology, AUUP, Noida
Visceral
Leishmaniasis
(VL)
or
kala-‐azar
is
a
major
health
problem
in
Indian
subcontinent
affecting
165.4
million
people
[1].
Moreover,
it
could
be
life-‐threatening
in
immunocompro-‐
mised
patients.
Existing
drugs
suffer
from
severe
toxicity,
long
treatment
regime
and
serious
side
effects.
High-‐throughput
screening
(HTS)
has
widely
been
considered
as
an
effective
tool
in
finding
new
lead
compounds.
Though,
a
number
of
novel
antiparasitic
agents
have
been
reported
from
plant
resources
however,
any
plant
derived
leishmanicidal
drug
or
drug
candi-‐
dates
have
not
been
reported
so
far.
Hence,
there
is
an
urgent
need
to
explore
potential
natu-‐
ral
resources
in
search
of
new
lead
compounds.
In
our
first
project,
we
screened
four
plants
including
Piper
longum,
Withania
somnifera,
Tinospora
cordifioia
and
Alstonia
scholaris
by
bio-‐
assay
guided
fractionation
(BAGF)
against
promastigotes
of
Leishmania
donovani
(DD8).
Max-‐
imum
bioactivity
and
least
cell
cytotoxicity
was
observed
with
n-‐Hex
fraction
of
P.
longum
(IC50
100
µg/mL).
Six
alkamides
(1,
3-‐7)
and
one
benzenoid
(2)
compound
[2,3]
including
three
novel
alkamides
were
identified
from
the
active
fraction
with
significant
leishmanicidal
activity
against
promastigotes
and
axenic
amastigotes
(Table
1).
Further
analysis
of
the
active
fraction
afforded
a
component
containing
piperine
and
piperine
isomers
(8a-‐8d)
in
the
pro-‐
portion
of
3.36:1
with
remarkably
high
activity.
In
addition
to
explore
the
binding
model
of
piperine
isomers
with
potential
drug
target,
pteridinere
ductase
1
(PTR1,
EC
1.5.1.33)
was
simulated
by
molecular
docking
[4].
A
fairly
good
agreement
between
experimental
data
and
molecular
docking
investigation
of
bioactive
and
inactive
compounds
was
observed.
To
iden-‐
tify
which
isomer
is
playing
the
key
role,
four
piperine
isomers
namely,
piperine
(2E,4E),
iso-‐
piperine
(2Z,4E),
isochavicine
(2E,4Z),
and
chavicine
(2Z,4Z)
were
studied
individually
against
LdPTR1.
The
result
demonstrated
that
isochavicine
exhibits
maximum
inhibitory
ef-‐
fect
(conformational
energy
of
56
kcal
mol-‐1)
by
forming
H-‐bonding,
hydrophobic
interaction
and
edge
to
face
π
stacking
interactions.
Presently,
preparation
of
synthetic
analogues
of
the
most
active
alkamides
is
in
process.
Compounds Promastigotes Amastigotes Cytotoxicity (%)
IC50± SD (µg/ml) IC50± SD (µg/ml) IC50± SDa (µg/ml)
Alkamide (1) 9.12± 0.34 2.81±0.14 5.05±0.64
Benzenoid (2) 11.85± 0.52 3.94±0.54 4.95±0.62
Alkamide (3) 14.90± 0.87 6.15±0.71 5.15±0.61
Alkamide (4) 14.15± 0.51 5.10±0.72 3.75±0.58
Alkamide (5) 14.85± 0.76 4.65±0.56 5.15±0.64
Alkamide (6) 14.21± 0.71 5.60±0.67 4.15±0.59
Alkamide (7) 14.85± 0.68 5.11±0.58 3.75±0.69
8a-‐8d 03.15±0.47 1.10±0.09 5.05±0.03
Miltefosine (positive control) 8.11± 0.36 4.37±0.51 25.15±0.63
a All compounds were examined in a set of triplicate experiments; 8a-‐ piperine, 8b-‐ isopiperine, 8c-‐isochavicine,
8d-‐chavicine
Table
1:
In
vitro
leishmanicidal
activity
of
compounds
1-‐8
on
promastigotes
and
axenic
amastigotes
of
L.donovani;
cell
cytotoxicity
of
the
compounds
on
J774A.1
cell
line
Acknowledgements:
The
authors
acknowledge
Department
of
Science
and
Technology
and
Department
of
Biotechnology
for
funding
and
Founder
President,
Amity
University
for
continuous
moral
support
and
encouragement.
Keywords: Leishmaniasis, Piper longum, LdPTR1, piperine isomers, alkamides
References:
[1] Toteja
GS,
Rajni
K.
Diseases
specific
documents
XII
plan:
Leishmaniasis.
Indian
Council
of
Medical
Research;
2014.
[2] Ghosal
S,
Deb
D,
Mishra
P,
Vishwakarma
R.
Leishmanicidal
compounds
from
the
fruits
of
Piper
longum.
Planta
Med
2012;
78:
906-‐908.
[3] Mishra
P,
Sinha
S,
Guru
SK
,
Bhushan
S
,
Vishwakarma
R.A
,
Ghosal
S.
Two
new
amides
with
cytotoxic
activity
from
the
fruits
of
Piper
longum.
J
Asian
Nat
Prod
Res
2011;
13:
143-‐148
[4] Tewatia
P,
Sahi
S,
Ghosal
S.
Leishmania
donovani
pteridine
reductase
1:
comparative
pro-‐
tein
modeling
and
protein-‐ligand
interaction
studies
of
the
leishmanicidal
constituents
isolated
from
the
fruits
of
Piper
longum.
J
Mol
Model
2012;
18:
5065-‐5073.
P437
Antiprotozoal
and
heme-‐binding
activity
of
13
aromatic
and
me-‐
dicinal
plants
from
Ahaggar,
Algeria
Kamel
Dali-‐Yahia2,
Sergio
Ortiz1,
Alexandre
Maciuk3,
Pedro
Vásquez-‐Ocmín3,
Laila
Salmen
Espindola4,
Sabrina
Boutefnouchet1
1
Laboratoire
de
Pharmacognosie,
UMR
CNRS
8638
COMETE,
Faculté
de
Pharmacie
de
Paris,
Université
Paris-‐Descartes,
Sorbonne
Paris
Cité,
4,
av.
de
l’Observatoire,
75006
Paris;
2
Département
de
Pharmacie,
Faculté
de
Médecine,
Université
de
Tlemcen,
13000,
Tlemcen,
Algeria;
3
Laboratoire
de
Pharmacognosie,
UMR
8076
CNRS
BioCIS,
Faculté
de
Pharmacie,
Université
Paris-‐Sud,
5
rue
J.-‐B.
Clément,
92296
Châtenay-‐
Malabry,
France;
4
Laboratório
de
Farmacognosia,
Universidade
de
Brasília,
Brasília,
Brazil
Thirteen
species
from
Ahaggar
area
were
selected
for
antiprotozoal
screening
on
the
basis
of
ethnopharmacological
uses
and/or
chemotaxonomy.
Among
the
selected
plants
10
Asterace-‐
ae,
2
Apiaceae
and
1
Lamiaceae
were
collected
in
the
Ahaggar
National
Park,
in
the
Southern
Sahara
[1,
2].
Both
antiplasmodial
activity
(P.
falciparum,
chloroquino-resistant FcB1/Colombia)
and
antileishmanial
activity
(promastigotes from L. amazonensis)
were
evaluated.
7
species
ex-‐
hibited
a
significant
antileshmanial
activity
(IC50
<
10
µg/ml)
and
8
species
showed
significant
antiplasmodial
activity
(IC50
≤
1
µg/l),
but
no
correlation
between
both
activities
was
ob-‐
served.
As
antiplasmodial
activity
could
be
associated
with
inhibition
of
heme
degradation
into
hemozoin,
with
subsequent
Plasmodium
death,
a
combined
heme-‐binding
assay
was
car-‐
ried
out
in
order
to
identify
stable
heme-‐adducts
[3].
Comparison
of
LC-‐MS
spectra
of
crude
extracts
with
or
without
heme
allowed
us
to
identify
m/z
values
of
compounds
involved
in
the
heme-‐adduct
formation.
Among
all
identified
adducts,
we
found
that
two
methoxyflavones
isolated
from
P.
monodiana
can
form
adducts
with
heme.
Antileishmania Antiplasmodial m/z values of com-‐
Plant Fraction activity activity pounds forming
(IC50 μg/ml) (IC50 μg/ml) heme-‐adducts
Ammodaucus leucotrichus (fruit), EtOAc 6.5 > 100 389 [M+H]+
Apiaceae MeOH -‐ 79.3 ± 7.7 381 [M+H]+
Anvillea radiata (aerial part), Aster-‐ 10.2 2.31 ± 0.86 277 [M+H]+, 287 [M+H]+,
EtOAc
aceae 344 [M+H]+
MeOH -‐ > 100 235 [M+H]+
Artemisia campestris ssp glutinosa 17.26 1.21 ± 0.16 315 [M+H]+, 330 [M+H]+,
EtOAc
(aerial part), Asteraceae 345 [M+H]+, 359 [M+Na]+
MeOH -‐ 0.72 ± 0.27 355 [M+H]+
Artemisia judaica ssp saharensis 8.93 1.13 ± 0.19 314 [M+H]+, 344 [M+H]+,
EtOAc
(aerial part), Asteraceae 420 [M+H]+
MeOH -‐ > 100 -‐
Cotula anthemoides (whole plant), EtOAc 6.59 78.4 ± 15.6 250 [M+H]+, 264 [M+H]+
Asteraceae MeOH -‐ 1.20 ± 0.32 250 [M+H]+, 395 [M+H]+
Deverra scoparia (aerial part) = Pitu-‐ 32.54 4.60 ± 1.50 239 [M+H]+, 357 [M+Na]+,
EtOAc
ranthos scoparia, Apiaceae 451 [M+Na]+
MeOH -‐ 20.4 ± 2.1 395 [M+Na]+
Deverra scoparia (root) EtOAc 7.27 > 100 301 [M+H]+, 451 [M+H]+
= Pituranthos scoparia, Apiaceae MeOH -‐ 0.99 ± 0.41 365 [M+H]+, 381 [M+H]+
Nauplius graveolens (aerail parts) = EtOAc 6.95 -‐ -‐
Asteriscus graveolens, Asteraceae MeOH -‐ > 100 235 [M+H]+
Pentzia monodiana (aerial parts), EtOAc 13.63 3.53 ± 1.08 359 [M+H]+, 389 [M+H]+
Asteraceae MeOH -‐ > 100 -‐
Pulicaria crispa (aerial parts) 14.93 36.2 ± 4.1 347 [M+H]+, 533 [M+H]+,
EtOAc
549 [M+H]+
MeOH -‐ 8.80 ± 1.80 156 [M+H]+, 235 [M+H]+
Pulicaria
incisa
(aerial
parts)
EtOAc
8.37
4.57
±
1.23
329
[M+H]+,
353
[M+H]+
MeOH
-‐
34.2
±
8.1
-‐
Rhaponticum
acaule
(root)
EtOAc
14
4.7
±
0.46
435
[M+H]+
MeOH
-‐
>
100
365
[M+H]+
Rhaponticum
acaule
(aerial
parts)
26
1.27
±
0.49
385
[M+H]+,
445
[M+H]+,
EtOAc
585
[M+H]+
MeOH
-‐
40.2
±
4.93
434
[M+H]+
Salvia
chudaei
(leaves),
Lamiaceae
EtOAc
-‐
>
100
329
[M+H]+,
369
[M+H]+
MeOH
-‐
64.2
±
40.5
399
[M+H]+
Acknowledgements:
Authors
thanks
the
ChemBioFight
European
funding
(FP7-‐people-‐2010-‐IRSES),
Al-‐
gerian
government
for
PROFAS
training
financial
support
and
Director
of
Ahaggar
National
Park.
Keywords: Ethnopharmacology, antiprotozoal, heme-‐binding, Ahaggar.
References:
[1] Ozenda
P.
Flore
et
végétation
du
Sahara,
Ed.
CNRS,
2004:
438.
[2] Hammiche
V,
Maiza
K.
Traditional
medicine
in
Central
Sahara:
Pharmacopoeia
of
Tassili
N’ajjer.
J
Ethnopharmacol
2006;
105:
358−367
[3] Muñoz-‐Durango
K,
Maciuk
A,
Harfouche
A,
Torijano-‐Gutiérrez
S,
Jullian
JC,
Quintin
J,
Spelman
K,
Mouray
E,
Grellier
P,
Figadère
B.
Detection,
characterization,
and
screening
of
heme-‐binding
molecules
by
mass
spectrometry
for
malaria
drug
discovery.
Anal
Chem
2012;
84:
3324−3329
P438
Chemical
characterization
of
Peganum
harmala
seeds
oil
with
evaluation
of
some
biological
activities
Khadhr
M1,
Bousta
M2,
Boukhira
S2,
Boukhchina
Sadok1
¹
Département
de
Biologie,
Faculté
des
Sciences
de
Tunis,
Université
de
Tunis
El
Manar,
2092
Tunisia,²
Unité
de
Toxico-‐Pharmacologie,
Agence
Nationale
des
Plantes
Médicinales
et
Aromatiques,
Université
Sidi
Mohamed
Ben
Abdellah,
Fès.
Maroc
Analysis
of
the
Peganum
harmala
seed
oil
by
GC,
GC-‐MS
and
HPLC
highlights
the
different
compounds
of
interest,
such
as
linoleic
acid
(62.05%
of
total
fatty
acids)
γ-‐
tocopherol
(573.66
µg/g
oil)
carotenoides
(2.4
mg/Kg
of
seed)
giving
it
some
biological
activities.
So
the
im-‐
munomodulatory
potential
of
the
cream
formulated
with
Peganum
harmala
seed
oil
was
eval-‐
uated
by
Flow
Cytometry.
An
immunosuppressive
effect
was
noted
on
monocytes
and
granu-‐
locytes
in
rats
[1].
This
effect
on
granulocytes
suggested
a
possible
anti-‐inflammatory
poten-‐
tial
for
this
plant
and
its
possible
eventual
use
in
the
treatment
of
inflammatory
diseases.
The
formulation
cream
reduced
inflammation,
five
hours
after
carrageenan
application,
compared
to
Diclofenac
at
1%
(60.4%;
45.65%;
respectively)
[2,3].
A
slight
potential
peripheral
analge-‐
sia
was
also
noted
in
plantar
test
in
treated
rats
with
cream
[4].
These
properties
are
due
mainly
to
its
richness
on
linoleic
acid,
γ-‐tocopherol
and
to
its
important
antioxidant
capacity.
Keywords:
Peganum
harmala,
bioactivities,
anti-‐inflammatory
References
[1] El
Hamsas
A,
Bousta
D,
Ouahidi
I,
Aarab
L.
Primary
pharmacological
screening
of
an
en-‐
demic
plant
from
the
Southern
Morocco
(Tetraena
gaetula
(Emb.
&
Maire)
Beier
&
Thu-‐
lin).
C
R
Biol
2010;
333:
736-‐743
[2] Hargreaves
K,
Dubner
R,
Brown
F,
Flores
C,
Joris
J.
A
new
and
sensitive
method
for
measur-‐
ing
thermal
nociception
in
cutaneous
hyperalgesia.
Pain
1988;
32:
77-‐88
[3] Posadas
I,
Bucci
M,
Roviezzo
F,
Rossi
A,
Parente
L,
Sautebin
L,
Cirino
G.
Carrageenan-‐
induced
mouse
paw
oedeme
is
biphasic,
age-‐weight
dependant
and
displays
differential
nitric
oxide
cyclooxygenase-‐2
expression.
Br
J
Pharmacol
2004;
142:
331-‐338
[4] Ayoola
GA,
Akpanika
GA,
Awobajo
FO,
Sofidiya
MO,
Osunkalu
VO,
Coker
HAB,
Odugbemi
TO.
Anti-‐inflammatory
properties
of
the
Allanblanckia
floribunda
(Guttiferae).
Bot
Res
Intl
2009;
2:
21-‐26
P439
Anti
freckles
plants
in
Iranian
Traditional
Medicine
Saeedeh
Ghafari1,
Shirin
Fahimi2,
Shamim
Sahranavard1,
2,
Ghazaleh
Heydarirad
3
1Traditional
Medicine
and
Materia
Medica
Research
Center,
Shahid
Beheshti
University
of
Medical
Sci-‐
ences,
No.19,
Tavaneer
Alley
,Vali-‐e-‐Asr
Ave.,
postal
code:1434875451,Tehran,
Iran,
2Department
of
Tra-‐
ditional
Pharmacy,
School
of
Traditional
Medicine,
Shahid
Beheshti
University
of
Medical
Sciences,
No.8,
Shams
Alley
,Vali-‐e-‐Asr
Ave.,
postal
code:1516745811,Tehran,
Iran,
3School
of
Traditional
Medicine,
Sha-‐
hid
Beheshti
University
of
Medical
Sciences,
No.8,
Shams
Alley
,Vali-‐e-‐Asr
Ave.,
postal
code:1516745811,Tehran,
Iran.
Freckles,
as
one
of
the
hyperpigmentary
disorders,
are
numerous
pigmented
spots
of
the
skin,
mainly
confined
to
the
face,
even
arms
and
back
which
may
cause
psychological
disturbances
[1-‐3].
One
of
the
several
methods
investigated
in
treatment
of
skin
hyperpigmentation
is
inhi-‐
bition
or
attenuation
of
tyrosinase
and
related
melanogenic
enzymes.
There
are
many
com-‐
mon
and
effective
whitening
agents
such
as
hydroquinone
and
its
derivatives,
kojic
acid
and
arbutin,
that
possess
a
tyrosinase
inhibitory
effect
[4].
Despite
their
benefit,
these
chemicals
would
cause
some
adverse
reactions
with
prolonged
exposures,
such
as
sensitization,
contact
dermatitis
and
erythema,
cytotoxic
and
mutagenic
effects
[3].
In
Iranian
Traditional
Medicine
(ITM),
which
dates
back
more
than
6000
years,
freckles
have
been
considered
as
well.
The
word
“namash”
was
the
term
used
by
ITM
scholars
to
indicate
freckles.
There
is
a
wide
range
of
plants
that
were
prescribed
by
Iranian
physicians
for
removing
freckles.
In
this
study,
we
reviewed
the
medicinal
plants
used
to
treat
freckles
via
topical
route
of
administration
in
three
famous
ITM
textbooks
including
Liber
Continent
(Rhazes),
Canon
of
Medicine
(Avicenna)
and
Makhzan
ul-‐Adwia
(Aghili).
Moreover,
the
plants
have
been
ordered
according
to
their
repetition
in
the
references.
Afterwards,
traditional
names
of
the
plants
were
matched
to
the
scientific
names
using
botanical
text
references.
Finally
a
substantial
search
of
scientific
data-‐
bases
such
as
‘Google
Scholar’
and
‘PubMed’
was
performed
for
the
plant
names
in
combina-‐
tion
with
the
terms
‘tyrosinase
inhibition’
to
find
the
herbals
with
tyrosinase
inhibitory
effect.
In
our
investigation,
twenty
plants
with
anti
freckles
effects
were
derived
from
ITM
textbooks.
Among
them,
Myrtus
communis,
Carthamus
tinctorius
and
Polygonum
hydropiper
have
shown
tyrosinase
inhibitory
effect
in
modern
scientific
research.
This
study
represents
a
list
of
herb-‐
al
medicines
for
future
studies
in
the
field
of
freckles.
Acknowledgements:
School
of
Traditional
Medicine,
Shahid
Beheshti
University
of
Medical
Sciences
is
acknowledged
for
library
support.
Keywords: Freckles, Iranian Traditional Medicine, herbal medicine, tyrosinase inhibition
References:
[1] He
PP,
Liang
YH,
Yang
S,
Yuan
WT,
Xu
ShJ,
Huang
W.
A
gene
for
freckles
maps
to
chromo-‐
some
4q32–q34.
J
Investig
Dermatol
2004;
122:
286-‐290
[2] Jang
K,
Chung
EC,
Choi
JH,
Sung
KJ,
Moon
KC,
Koh
JK.
Successful
removal
of
freckles
in
Asian
skin
with
a
Q-‐switched
alexandrite
laser.
Dermatol
Surg
2000;
26:
231-‐234
[3] Adhikari
A,
Devkota
HP,
Takano
A,
Masuda
K,
Nakane
T,
Basnet
P,
Skalko-‐Basnet
N.
Screening
of
Nepalese
crude
drugs
traditionally
used
to
treat
hyperpigmentation:
in
vitro
tyrosinase
inhibition.
Int
J
Cosmet
Sci
2008;
30:
353-‐360
[4] Ortonne
JP,
Bissett
DL.
Latest
insights
into
skin
hyperpigmentation.
J
Investig
Dermatol
Symp
Proc
2008;
13:
10-‐14
P440
Alkaloids
of
Narcissus
poeticus
cv.
Pink
Parasol
and
their
biologi-‐
cal
activity
Šafratová
Marcela1,
Hošťálková
Anna1,
Opletal
Lubomír1,
Kuneš
Jiří2,
Cahlíková
Lucie
1
1
ADINACO
Research
Group,
Department
of
Pharmaceutical
Botany
and
Ecology,
Faculty
of
Pharmacy
in
Hradec
Králové,
Charles
University
in
Prague,
500
05
Hradec
Králové,
Czech
Republic,
2
Department
of
Inorganic
and
Organic
Chemistry,
Faculty
of
Pharmacy
in
Hradec
Králové,
Charles
University
in
Prague,
500
05
Hradec
Králové,
Czech
Republic
Alzheimer´s
disease
(AD)
is
known
as
one
of
the
most
common
neurodegenerative
diseases.
The
main
characteristic
sign
of
AD
is
progressive
and
irreversible
loss
of
neurons.
AD
is
also
characterized
by
decreased
levels
of
neurotransmitter
acetylcholine
(Ach)
in
the
cortex,
which
is
hydrolyzed
mostly
by
acetylcholinesterase
(AChE)
in
healthy
brain.
On
the
other
hand,
in
late
AD
stages
butyrylcholinesterase
(BuChE)
is
the
main
hydrolysing
enzyme
as
its
content
will
increase
by
up
to
90%
in
comparison
to
normal
state.
BuChE
cleaves
ACh
in
a
similar
way
as
AChE
to
terminate
its
physiological
action
[1].
Plants
of
Amaryllidaceae
species
are
important
for
producing
specific
compounds
that
are
known
as
Amaryllidaceae
alkaloids.
These
alkaloids
have
interesting
physiological
effects
such
as
antitumor,
antiviral,
antimalarial
and
acetylcholinesterase
activity.
Alkaloidal
extracts
of
some
Narcissus
species
have
been
tested
for
their
inhibiting
effects
on
acetylcholinesterase
and
butyrylcholinesterase
and
alkaloid
pattern.
Interesting
biological
activities
were
demon-‐
strated
by
extracts
of
N.
poeticus
cv.
Pink
Parasol
IC50
HuBuChE
=
3.3
±
0.5
µg/ml
(IC50
HuAChE
=
191.3
±
20.2
µg/ml)
and
N.
poeticus
cv.
Gigantic
Star
IC50
HuAChE
=
3.2
±
0.6
µg/ml
(IC50
HuBuChE
=
16.2
±
1.0
µg/ml).
Due
to
the
interesting
inhibition
activity
and
the
presence
of
lycorine
and
homolycorine
type
of
alkaloids
in
the
crude
extracts,
a
phytochemical
study
was
performed
on
N.
poeticus
cv.
Pink
Parasol.
Fourty-‐two
alkaloids
were
determined
by
GC/MS,
thirty
of
them
were
identified
from
their
mass
spectra,
retention
times
and
indexes.
Many
of
them
are
new
structures
that
have
not
been
isolated
yet.
Their
biological
activities
are
also
unknown.
Acknowledgements:
SVV-‐2016-‐
260292
Keywords: Amaryllidaceae, Alzheimer´s disease, GC/MS, AChE, BuChE
References:
[1] Park
SY.
Potential
therapeutic
agents
against
Alzheimer´s
disease
from
natural
sources.
Arch
Pharmacal
Res
2010;
33:
1589-‐1609
P441
Bioactive
labdane
diterpenoids
from
Salvia
leriifolia
Samad
Nejad
Ebrahimi1,
Mahdi
Moridi
Farimani1,
Akram
Taleghani1,
Abbas
Aliabadi2,
Atousa
Aliahmadi3,
Mohammad
Ali
Esmaeili3,
Nazanin
Namazi
Sarvestani4,
Hamid
Reza
Khavasi5,
Martin
Smieško6,
Matthias
Hamburger7
1
Department
of
Phytochemistry,
Medicinal
Plants
and
Drugs
Research
Institute,
Shahid
Beheshti
Univer-‐
sity,
G.
C.,
Evin,
Tehran,
Iran;
2
Khorasan
Razavi
Agricultural
and
Natural
Resources
Research
Center,
Sabzevar
Branch,
Sabzevar,
Khorasan
Razavi,
Iran;
3
Department
of
Biology,
Medicinal
Plants
and
Drugs
Research
Institute,
Shahid
Beheshti
University,
G.C.,
Evin,
Tehran,
Iran;
4
Department
of
Animal
Biology,
School
of
Biology,
College
of
Science,
University of
Tehran,
Tehran,
Iran;
5
Department
of
Chemistry,
Fac-‐
ulty
of
Chemistry,
Shahid
Beheshti
University,
G.C.,
Evin,
Tehran,
Iran;
6
Division
of
Molecular
Modeling,
University
of
Basel,
Klingelbergstrasse
50,
4056
Basel,
Switzerland;
7
Division
of
Pharmaceutical
Biology,
University
of
Basel,
Klingelbergstrasse
50,
4056
Basel,
Switzerland.
Fractionation
of
an
n-‐hexane
extract
of
the
aerial
parts
of
Salvia
leriifolia
as
endemic
Iranian
plant
led
to
the
isolation
of
two
new
(1,
2)
and
two
known
(3,
4)
labdane
diterpenoids,
to-‐
gether
with
three
other
known
compounds.
The
structures
were
established
by
a
combination
of
1D
and
2D
NMR,
and
HRESIMS.
The
structures
of
1
and
3
were
confirmed
by
single-‐crystal
X-‐ray
analysis.
The
absolute
configuration
of
1-‐4
was
established
by
electronic
circular
di-‐
chroism
(ECD)
spectroscopy.
Compounds
1-‐4
were
evaluated
for
their
cytotoxic
activities
against
MCF-‐7
human
breast
cancer
cells.
Labdanes
3
and
4
were
in
addition
tested
against
MDA-‐MB231
human
breast
cancer,
and
DU-‐145
human
prostate
cancer
cell
lines.
Compound
4
showed
IC50
values
of
25,
50,
and
50
µg/mL
against
MCF-‐7,
MDA-‐MB231,
and
DU-‐145
cells,
respectively.
Compounds
1-‐4
were
tested
for
activity
against
Gram-‐positive
(Staphylococcus
aureus)
and
Gram-‐negative
(Escherichia
coli)
bacteria.
Compound
3
showed
a
MIC
of
64
μg/mL
against
methicillin
resistant
S.
aureus
(MRSA).
Acknowledgements:
We
are
grateful
to
Shahid
Beheshti
University
Research
Council
for
financial
support
of
this
work.
Keywords: Salvia leriifolia, lamiaceae, diterpene, absolute configuration, antibacterial activi-‐
ty, cytotoxicity
P442
Resistance
modulatory
and
efflux
pump
inhibitory
activities
of
Kaempferia
galanga
rhizomes
Sandra
Prasch1,
Britta
Fimbinger1,
Franz
Bucar1,2
1
Institute
of
Pharmaceutical
Sciences,
2
Department
of
Pharmacognosy,
University
of
Graz,
Universi-‐
tätsplatz 4, 8010 Graz, Austria
Antibiotic
resistance
represents
an
increasing
problem
for
public
health
[1].
Bacteria
rapidly
adapt
to
environmental
conditions
and
so
the
application
of
large
amounts
of
antibiotics
(in
human
and
livestock
farming)
and
non-‐compliance
further
pushes
the
development
of
re-‐
sistant
strains.
The
presence
of
efflux
pumps
on
the
cell
membrane
is
one
way
how
bacteria
can
become
resistant.
These
pumps
are
capable
of
effluxing
toxic
compounds
from
the
cyto-‐
plasm.
Therefore,
a
possible
starting
point
to
reverse
these
developments
is
the
exploration
of
efflux
pump
inhibitors
[2].
Plants
from
the
family
of
Zingiberaceae
proved
to
be
a
good
source
for
efflux
pump
inhibitory
compounds
[3].
In
this
study
the
rhizomes
of
Kaempferia
galanga
L.
were
studied
for
the
presence
of
such
components.
Three
rhizome
extracts
were
manufactured
and
tested
against
Mycobacterium
smegmatis
mc²
155.
Especially
the
hexane
extract
revealed
promising
re-‐
sistance
modulatory
activities
bearing
a
modulation
factor
(MF)
of
64
with
ethidium
bromide
(EtBr)
and
an
even
higher
MF
of
≥
256
when
tested
with
rifampicin.
Due
to
these
promising
values
this
extract
was
bioassay-‐guided
fractionated
in
order
to
reveal
the
most
active
frac-‐
tions.
GC-‐MS
analysis
revealed
the
presence
of
ethyl-‐p-‐methoxycinnamate
in
fractions
with
the
highest
MF
of
32.
For
this
reason
an
accumulation
assay
using
EtBr
as
substrate
was
per-‐
formed
showing
superior
accumulation
behaviour
of
ethyl-‐p-‐methoxycinnamate
compared
to
verapamil
and
a
concentration
dependent
EtBr
accumulation.
Acknowledgements:
University
of
Graz
is
gratefully
acknowledged
for
providing
a
PhD
scholarship
(“For-‐
schungsstipendium
für
Doktorandinnen”)
to
Sandra
Prasch.
Keywords: Efflux pump inhibitor, Kaempferia galanga, Mycobacterium smegmatis mc² 155
References:
[1] World
Health
Organisation.
Antimicrobial
resistance
global
report
on
surveillance
2014.
Geneva:
WHO,
2014.
[2] Fernández
L,
Hancock
REW.
Adaptive
and
mutational
resistance:
role
of
porins
and
efflux
pumps
in
drug
resistance.
Clin
Microbiol
Rev
2012;
25:
661-‐681
[3] Groeblacher
B,
Maier
V,
Kunert
O,
Bucar
F.
Putative
mycobacterial
efflux
inhibitors
from
the
seeds
of
Aframomum
melegueta.
J
Nat
Prod
2012;
75:
1393-‐1399
P443
Novel
polyketides
isolated
from
cultures
of
an
endophytic
fungus,
Annulohypoxylon
truncatum
Wei
Li1,
Changyeol
Lee2,
Sung
Hee
Bang2,
Soonok
Kim3,
Xuikui
Xia4,
Sang
Hee
Shim2
1
School
of
Biotechnology,
Yeungnam
University,
Gyeongsan
712-‐745,
South
Korea,
2
College
of
Pharmacy,
Duksung
Women’s
University,
Seoul
132-‐714,
South
Korea,
3
National
Institute
of
Biological
Resources,
Incheon
404-‐708,
South
Korea,
4
Key
Laboratory
for
Applied
Microbiology
of
Shandong
Province,
Biotech-‐
nology
Center
of
Shandong
Academy
of
Sciences,
Jinan
250014,
P.
R.
China,
Endophytes
are
microbes
that
colonize
living,
internal
tissues
of
plants
without
causing
any
immediate,
negative
effects
[1].
The
ecological
roles
and
chemical
interactions
of
endophytes
are
currently
under
investigation,
particularly
in
relation
to
their
host
plants.
It
is
known
that
endophytes
are
a
rich
and
reliable
source
of
genetic
diversity
and
may
represent
previously
undescribed
species
[2,3].
Novel
microbes
(as
defined
at
the
morphological
and
/or
molecular
level)
often
are
associated
with
novel
natural
products.
In
an
effort
to
investigate
natural
compounds
with
intriguing
structures
from
endophytic
fungi,
we
selected
plants
from
a
dis-‐
tinct
environmental
setting
which
could
be
a
promising
source.
Several
endophytes
were
iso-‐
lated
from
the
stems
of
aquatic
reed
plants.
Among
them,
a
strain
of
endophyte,
Annulohy-‐
poxylon
truncatum
(JS540),
was
isolated
from
barks
of
reed
plants
and
cultivated
on
a
large
scale.
And
they
were
extracted
with
ethyl
acetate,
which
were
subjected
to
a
series
of
chroma-‐
tographic
methods,
leading
to
the
isolation
of
eight
secondary
metabolites
including
six
new
compounds
(1:
7.8
mg,
2:
4.5
mg,
3:
3.5
mg,
5:
5.6
mg,
6:
4.5
mg,
and
8:
4.6
mg).
The
isolated
compounds
were
identified
by
analysis
of
spectroscopic
methods
such
as
1D-‐,
2D-‐NMR,
and
MS
as
shown
in
the
following
figure.
Acknowledgements:
This
work
was
supported
by
Priority
Research
Centers
Program
through
the
Nation-‐
al
Research
Foundation
of
Korea
(NRF)
funded
by
the
Ministry
of
Education,
Science
and
Technolo-‐
gy(2016R1A6A1A03007648)
and
Korea
Research
Foundation
(NRF-‐2014K2A2A2000728
and
NRF-‐
2015R1D1A1A01057914).
Keywords: Endophyte, polyketide, Annulohypoxylon truncatum
References:
[1] Strobel
G,
Daisy
B,
Castillo
U,
Harper
J.
Natural
products
from
endophytic
microorganisms
J
Nat
Prod
2014;
67:
257-‐268
[2] Smith
SA,
Tank
DC,
Boulanger
LA,
Bascom-‐Slack
CA,
Eisenman
K,
Kingery
D,
Babbs
B,
Fenn
K,
Greene
JS,
Hann
BD,
Keehner
J,
Kelley-‐Swift
EG,
Kembaiyan
V,
Lee
SJ,
Li
P,
Light
DY,
Lin
EH,
Ma
C,
Moore
E,
Schorn
MA,
Vekhter
D,
Nunez
PV,
Strobel
GA,
Donoghue
MJ,
Strobel
SA.
Bioactive
endophytes
warrant
intensified
exploration
and
conservation.
PLoS
One
2008;
3:
e3052
[3] Newman
DJ,
Cragg
GM.
Endophytic
and
epiphytic
microbes
as
"sources"
of
bioactive
agents.
Front
Chem
2015;
3:
34
P444
Anti-‐fatigue
effects
of
Rhodiola
rosea
extract
Sang
Yoon
Choi
Korea
Food
Research
Institute,
Seongnam
463-‐746,
Republic
of
Korea
Regular
exercise
is
known
to
help
protect
and
alleviate
hypertension,
stroke,
cardiovascular
disease,
diabetes,
hyperlipidemia,
and
cancer
[1,2].
However,
strenuous
exercise
causes
ex-‐
cessive
production
of
reactive
oxygen,
lipid
peroxides,
and
lactic
acid,
which
can
damage
mus-‐
cle
tissues
[3-‐5].
Rhodiola
rosea
L.
(Crassulaceae)
is
a
perennial
plant
which
grows
in
the
al-‐
pine
regions
of
Europe
and
Asia
[6].
This
study
was
conducted
to
examine
the
protective
ef-‐
fects
of
fermented
Rhodiola
rosea
extract
against
fatigue
and
exercise
stress.
As
a
result,
Rho-‐
diola
rosea
extract
significantly
inhibited
L6
muscle
cell
death
(57.4%
at
50
ppm)
and
ATP
reduction
(70.0%
at
50
ppm)
caused
by
H2O2
damage.
In
addition,
the
oral
administration
of
Rhodiola
rosea
extract
in
mice
improved
treadmill
running
time
by
43.9%
and
blood
profiles
of
glutamic
oxaloacetic
transaminase,
glutamic
pyruvic
transaminase,
lactate
dehydrogenase,
creatinine,
lactate
when
compared
to
distilled
water
by
106.9%,
68.0%,
29.2%,
100.0%,
50.0%
respectively.
Given
the
above
results,
it
is
considered
that
Rhodiola
rosea
extract
effectively
protects
against
fatigue
caused
by
strenuous
exercise.
Acknowledgements:
This
study
was
supported
by
research
program
of
Korea
food
research
institute
Keywords: Rhodiola rosea, exercise, anti-‐fatigue effect
References:
[1] Bassuk
SS,
Manson
JE.
Epidemiological
evidence
for
the
role
of
physical
activity
in
reduc-‐
ing
risk
of
type
2
diabetes
and
cardiovascular
disease.
J
Appl
Physiol
2005;
99:
1193-‐
1204
[2] Roberts
CK,
Barnard
RJ.
Effects
of
exercise
and
diet
on
chronic
disease.
J
Appl
Physiol
2005;
98:
3-‐30
[3] Jenkins
RR.
Excercise,
oxidative
stress,
and
antioxidants.
Int
J
Sport
Nutr
1993;
3:
356-‐375
[4] Hyun
KY.
An
association
of
changed
levels
of
inflammatory
markers
with
hematological
factors
during
one-‐time
aerobic
exercise
in
twenty-‐aged
young
men.
J
Life
Sci
2009;
19:
1658-‐1665
[5] Moxnes
JF,
Sandbakk
Ø.
The
kinetics
of
lactate
production
and
removal
during
whole-‐body
exercise.
Theor
Biol
Med
Model
2012;
9:
7
[6] Chan
SW.
Panax
ginseng,
Rhodiola
rosea
and
Schisandra
chinensis.
Int
J
Food
Sci
Nutr
2012;
63:
75-‐81
P445
Vaccinium
cereum
leaves,
Tahitian
berry,
a
common
endemic
plant
with
α-‐glucosidase
inhibitory
activity
Sarah
Benayad1,
Estelle
Adam1,
Stéphanie
Soulet1,
Isabelle
Bombarda2,
Cécile
Debitus3,
Taivini
Teai1
1UMR
241
EIO,
University
of
French
Polynesia,
PoBox
6570
–
98702
Faaa
–
Tahiti,
French
Polynesia,
2IMBE-‐UMR
7263,
Aix-‐Marseille
University,
St
JEROME-‐
Avenue
Escadrille
Normandie
Niemen
-‐13013
Marseille France, 3UMR 241 EIO, IRD, PoBox 529-‐98713 Papeete-‐ Tahiti, French Polynesia
Diabetes
mellitus
is
a
major
cause
of
death
worldwide
and
affected
9%
of
the
population
in
2014
[1].
In
French
Polynesia,
type
2
diabetes
is
the
second
noncommunicable
disease
after
high
arterial
blood
pressure,
and
10%
of
the
population
is
concerned
[2].
The
extent
of
this
disease
encourages
the
search
for
new
treatments.
French
Polynesia,
one
well-‐known
world
biodiversity
“hot
spot”,
is
an
overseas
French
territory.
Its
terrestrial
vascular
flora
is
original,
with
54%
endemism
[3].
Studies
on
the
bioactivity
and
phytochemistry
of
those
plants
are
very
scarce.
Given
that
α-‐glucosidase
plays
an
important
role
in
the
regulation
of
type
2
diabe-‐
tes,
we
have
undertaken
a
search
for
an
α-‐glucosidase
inhibitor
among
those
endemic
plants.
Vaccinium
cereum
var.
cereum
was
selected
for
its
strong
in
vitro
α-‐glucosidase
inhibitory
ac-‐
tivity.
This
species
has
never
been
studied
from
a
phytochemical
point
of
view
and
such
activi-‐
ty
or
therapeutic
traditional
use
never
been
reported.
A
bio-‐guided
phytochemical
study
was
carried
out
on
V.
cereum
var.
cereum
using
column
chromatography
and
HPLC.
Various
com-‐
pounds
were
isolated
and
characterized
by
NMR,
including
the
active
compounds
ursolic
acid
(IC50=23.87µM),
oleanolic
acid
(IC50=9.20µM)
and
(E)-‐4-‐methoxycinnamic
acid
(IC50=
42.10µM).
The
strong
activities
observed
were
compared
to
the
standard
1-‐deoxynojirimycin
(IC50=30.64µM).
These
compounds
were
already
known
to
possess
inhibitory
activity
against
α-‐glucosidase
[4].
In
French
Polynesia,
there
are
4
varieties
of
V.
cereum.
The
3
varieties:
var.
adenandrum
(Decne)
F.Br.,
var.
cereum
(L.f.)
G.
Forst
and
var.
pubiflorum
Skottsb
are
common
and
have
shown
comparable
inhibitory
activities
against
α-‐glucosidase.
The
fourth
one,
var.
raiateense
(J.W.Moore)
M.L.
Grant
is
considered
as
an
endangered
category
(IUCN
red
list)
and
could
not
be
included
in
this
study.
V.
cereum
represents
a
real
health
benefit
and
could
be
used
by
local
people
to
improve
blood
glucose
control.
Acknowledgements:
French
Polynesia
research
delegation
and
ED
469
for
financial
support
Keywords: Vaccinium cereum, α-‐glucosidase inhibitory, phytochemical study.
References:
[1] WHO.
Global
status
report
on
noncommunicable
diseases
2014.
World
Health
Organiza-‐
tion,
2014;
xi-‐xvii,
ISBN
:
978-‐9-‐241567-‐85-‐4
(http://apps.who.int/iris/bitstream/10665/148114/1/9789241564854_eng.pdf?ua=1)
[2] Bertrand
S,
Paupier
A,
Tuheiava
M.
Campagne
2013
de
lutte
contre
le
diabète
de
type
2
en
Polynésie
française
:
«
Le
diabète,
je
peux
l’éviter
»
BISES
2014;
11:
1-‐4
[3] UICN
France,
MNHN
&
DIREN
Polynésie
française.
La
Liste
rouge
des
espèces
menacées
en
France
-‐
Chapitre
Flore
vasculaire
endémique
de
Polynésie
française.
Paris,
France,
2015:
1-‐19.
ISBN
:
978-‐2-‐918105-‐55-‐8.
[4] Benalla
W,
Bellahcen
S,
Bnouham
M.
Antidiabetic
medicinal
plants
as
a
source
of
alpha-‐
glucosidase
inhibitors.
Curr
Diabetes
Rev
2010;
6:
247-‐254.
P446
New
flavonoid
from
Polygonum
equesitiform
Sm.
and
antioxidant
activity
of
its
extract
Sayed
A.
El-‐Toumy,
Josline
Y.
Salib,
Nabila
H.
Shafik,
Asmaa
Sayed
Chemistry of Tannins Department, National Research Center, 12622 Dokki, Cairo, Egypt
In
the
early
nineties
the
presence
of
flavonoids
in
herbal
began
to
attract
the
attention
of
a
number
of
researchers,
as
a
result
of
their
biological
and
physiological
importance
[1].
The
present
study
deals
with
the
isolation
and
identification
of
flavonoids
from
Polygonum
equesi-‐
tiform
and
evaluation
of
antioxidant
activity
of
the
extract.
The
aqueous
alcoholic
extract
(MeOH:
H2O,
8:2)
of
Polygonum
equesitiform
aerial
parts
was
subjected
to
extensive
repeated
column
chromatography
on
polyamide,
and
Sephadex
LH-‐20
resulted
in
a
new
flavonoid
named
3,
5,
7,
2ʼ,
5-‐pentahydroxyflavone
3-‐
glucoside
as
well
as
quercetin
3-‐O-‐β-‐
glucopyranoside,
quercetin
3-‐O-‐β-‐glucuronide
6ʼʼ-‐metyl
ester,
quercetin
7-‐O-‐β-‐
glucopyranoside,
gallic
acid,
p-‐methoxy
gallic
acid,
gallic
acid
methyl
ester,
myricetin
and
quercetin.
Structures
of
the
isolated
compounds
were
established
by
chromatography,
UV,
HRESI-‐MS
and
1D/2D
1H/13C
NMR
spectroscopy.
The
radical
scavenging
activity
of
the
extract
was
measured
spectrophotometrically,
using
DPPH
radical.
The
ED50
(37.45
µM)
of
the
extract
was
compared
with
standard
antioxidants
as
vitamin
C.
Keywords:
Polygonum
equesitiform,
new
flavonoid,
antioxidant
activity
References:
[1] Havsteen
B.
The
biochemistry
and
medical
significance
of
the
flavonoids.
Pharmacol
Ther
2002;
96:
67-‐202
P447
Isolation
of
flavonoids
from
Acacia
albida
leaves
and
evaluation
of
antihyperglycaemic
effects
of
its
extract
Sayed
A.
El-‐Toumy1,
Ahmed
H.
Gaara2,
Abdel
Razik
H.
Farrag3,
Nadia
M.
Ahmed4
1
Chemistry
of
Tannins
Department,
2
Chemistry
of
Natural
compounds,
3
Pathology
Department,
4
Medical
Biochemistry Department, National Research Center, 12622 Dokki, Cairo, Egypt
Diabetes mellitus is a chronic disease caused by inherited and/or acquired deficiency in pro-‐
duction of insulin by the pancreas, or by the ineffectiveness of the insulin produced. The pre-‐
sent study deals with the isolation and identification of the flavonoid constituents from Acacia
albida leaves and evaluation of antihyperglycaemic effect of an aqueous methanol extract. The
aqueous alcoholic extract (MeOH:H2O, 7:3) of A. albida leaves was subjected to extensive re-‐
peated column chromatography on polyamide, cellulose and Sephadex LH-‐20, resulting in
isolation of quercetin-‐3-‐O-‐β-‐ glucopyranoside, kaempferol 3-‐O-‐β-‐glucopyranoside, quercetin-‐
3-‐O-‐α-‐L-‐rhamnopyranoside, kaempferol 3-‐O-‐α-‐L-‐rhamnopyranoside, quercetin-‐3-‐O-‐α-‐L -‐
arabinoside, quercetin and kaempferol. The structures of the isolated compounds were eluci-‐
dated on the basis of spectral analysis (UV, HRESI-‐MS and 1D/2D 1H/13C NMR spectroscopy).
The extract was tested in two dose levels 100 and 200 mg/kg (orally using an intra-‐gastric)
for 4 weeks. At baseline, fasting blood glucose levels of streptozotocin-‐diabetic rats were ele-‐
vated to (378.16±25.27 mg/dl) compared to control value. Treatment with the plant extracts
significantly reduced blood glucose in a dose dependent manner in treated rats. The Acacia
albida extract also significantly decreased total cholesterol (120.50±3.26; 102.33±1.56
mg/dl), triglyceride (144.66±2.74; 113.33±3.48 mg/dl) and increased HDL (52.50±1.66;
55.16±2.00 mg/dl) in lower and higher dose compared with streptozotocin-‐treatment alone.
The extract failed to produce hyperglycemic activity in control rats. The chemical constituents
of the plant, especially phenolics and other compounds present in the plant may be involved
in the observed hypoglycemic effect of the plant extract [1]. The results show that the oral
administration of A. albida leaf extract on the diabetic state reduced hyperglycemia in this
model.
Keywords: Acacia albida, flavonoids, anti-‐diabetic
References:
[1] Resurreccion-‐Mago M, Villase I, Harada N, Monde K. Antihyperglycaemic flavonoids from
Syzygium samarangense (Blume) Merr. and Perry. Phytother Res 2005; 19: 246-‐251
P448
Physiological
activities
of
Rubus
crataegifolius
fruits
in
Korea
Hyeusoo
Kim,
Moon-‐Sup
Kim,
Sugwang
Lee,
Uk
Lee,
Sea-‐Hyun
Kim
Special-‐purpose Trees Division, National Institute of Forest Science, Suwon 16631, Republic of Korea
Rubus
crataegifolius
Bunge
is
a
kind
of
raspberry,
which
is
native
to
East
Asia.
The
fruit
of
R.
crataegifolius
is
an
aggregate
fruit
1
cm
in
diameter,
made
up
of
numerous
drupelets
and
is
used
as
food
[1].
This
study
was
carried
out
to
analyze
the
bioactive
components,
antioxidant
activities
of
R.
crataegifolius
grown
in
12
different
regions
in
Korea.
The
total
phenolic
con-‐
tents
ranged
from
1.79-‐3.85mg/g
which
was
measured
by
Folin-‐denis
method
[2]
and
the
total
flavonoid
contents
ranged
of
1.00-‐1.97mg/g
which
was
measured
by
Davis
method
[3].
The
total
anthocyanin
contents
ranged
of
7.64-‐47.9mg/100g
and
vitamin
C
contents
ranged
of
0.33-‐0.59mg/g
which
was
measure
by
AOAC
method
[4].
DPPH
free
radical
scavenging
activi-‐
ty
ranged
of
32.81-‐67.79%
and
ABTS
radical
scavenging
activity
ranged
of
15.99-‐56.29%
at
500ppm.
Our
data
revealed
that
the
region
of
R.
crataegifolius
origin
greatly
influenced
the
phytochemical
contents
and
antioxidant
activity
of
R.
crataegifolius,
which
might
help
for
the
selection
and
validation
of
the
most
productive
cultivars
in
the
functional
food
industries.
Keywords:
polyphenol,
flavonoid,
anthocyanin,
vitamin
C,
antioxidant
activity,
Rubus
cra-‐
taegifolius
References:
[1] Bobinaitė
R.,
Viškelis
P,
Venskutonis
PR.
Variation
of
total
phenolics,
anthocyanins,
ellagic
acid,
and
radical
scavenging
capacity
in
various
raspberry
(Rubus
spp.)
cultivars.
Food
Chem
2012;
132:
1495-‐1501
[2] Gulcin
I,
Sat
IG,
Beydemir
S,
Elmastas
M,
Kufrevioglu
OI.
Comparison
of
antioxidant
activity
of
clove
(Eugenia
caryophylata
Thunb.)
buds
and
lavender
(Lavandula
stoechas
L.).
Food
Chem
2004;
87:
393-‐400
[3] Abeysinghe
DC,
Li,
X,
Sun,
S,
Zhang
W,
Zhou
C,
Chen
K.
Bioactive
compounds
and
antioxidant
capacities
in
different
edible
tissues
of
citrus
fruit
of
four
species.
Food
Chem
2007;
104:
1338-‐1344
[4] Lee
J.
Determination
of
total
monomeric
anthocyanin
pigment
content
of
fruit
juices,
bev-‐
erages,
natural
colorants,
and
wines
by
the
pH
differential
method:
Collaborative
Study.
J
AOAC
Int
2005;
88:
1269-‐1278
P449
Antioxidant
activities
and
useful
components
of
Korean
Walnut
(Juglans
sinensis)
kernels
Youngki
Park,
Sea-‐Hyun
Kim,
Jae-‐Hee
Kim
Special-‐purpose
Trees
Division,
National
Institute
of
Forest
Science,
Suwon
16631,
Republic
of
Korea
Walnut
(Juglans
sinensis
Dode,
Juglandaceae)
kernels
contain
high
levels
of
phenolic
com-‐
pounds
which
show
antioxidant
properties
[1].
The
aims
of
this
study
were
(a)
to
extract
the
phenolic
fractions
from
walnut
kernels,
and
to
examine
their
antioxidant
potentials,
and
(b)
to
determine
the
total
phenolics
for
evaluating
the
correlation
between
antioxidant
potential
and
total
phenolics.
Walnut
kernels
were
collected
from
the
walnut
trees
grown
in
the
Korea
Forest
Research
Institute
(Suwon).
Dried
walnut
kernels
were
finely
ground,
extracted
twice
with
ethanol
(EtOH)
and
then
evaporated
to
give
the
crude
extract.
The
crude
extracts
of
wal-‐
nut
kernels
were
successively
partitioned
with
various
organic
solvents
in
the
order
of
di-‐
chloromethane
(DCM),
ethyl
acetate
(EtOAc)
and
butanol
(Bu).
From
in
this
study,
we
could
suggest
that
ethyl
acetate
fraction
has
the
highest
antioxidant
and
total
phenolic
content
among
others.
Total
phenolics
in
walnut
EtOH
extract
(WE),
DCM
fraction
(WD),
EtOAc
frac-‐
tion
(WEA),
and
Bu
fraction
(WBu)
were
83.9
±
4.4,
43.6
±
14.3,
360.4
±
8.8,
and
189.3
±
19.9
mg/g,
respectively,
while
the
free
radical
scavenging
activities
of
the
extract/fractions
were
70.5
±
7.3
(WE),
31.0
±
6.4
(WD),
90.1
±
0.5
(WEA),
and
90.0
±
1.3%
(WBu)
at
100
µg/ml.
A
linear
correlation
was
also
found
between
free-‐radical
scavenging
activity
and
total
phenolics.
From
the
correlation
data,
we
can
also
conclude
that
the
antioxidant
activity
of
walnut
kernels
is
also
closely
correlated
with
the
total
phenolics.
Keywords:
antioxidant
activity,
vitamin
C,
Juglans
sinensis,
walnut
kernels
References:
[1] Diana
OL,
Maestri
DM,
Perello
M.
Matinez
ML.
Phenolic
from
Walnut
(Juglans
regia
L.)
ker-‐
nels:
Antioxidant
activity
and
interactions
with
proteins.
Food
Chem
2008;
107:
607-‐612
P450
Phytochemical
analysis
of
aerial
parts
of
Persicaria
bistorta
(L.)
Samp.
and
the
evaluation
of
the
anti-‐inflammatory
potential
of
rare
malonylated
flavonols
Urszula
Klimczak1,
Marta
Woźniak1,
Michał
Tomczyk2,
Sebastian
Granica1
1
Department
of
Pharmacognosy
and
Molecular
Basis
of
Phytotherapy,
Faculty
of
Pharmacy,
Medical
University
of
Warsaw,
Banacha
1,
02-‐097
Warsaw,
Poland,
2
Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Medical
University
of
Białystok,
Mickiewicza
2A,
15-‐230
Białystok,
Poland
Persicaria
bistorta
(L.)
Samp.
commonly
known
as
common
bistort
is
a
perennial
herbaceous
plant
belonging
to
the
Polygonaceae
family
and
it
is
native
to
Europe
and
Asia.
Leaves
of
P.
bistorta
were
consumed
by
people
in
Europe
and
Asia.
In
England,
especially
in
West
York-‐
shire,
leaves,
young
shoots
or
aerial
parts
of
bistort
were
used
as
an
ingredient
of
dock
pud-‐
ding
[1,2].
Up
to
date
little
is
known
about
the
chemical
composition
of
aerial
parts
of
com-‐
mon
bistort
[3].
The
UHPLC-‐DAD-‐MS
system
was
set
for
the
qualitative
analysis
of
extracts
from
aerial
parts
of
P.
bistorta.
In
total
around
40
compounds
were
detected
and
character-‐
ized.
Additionally,
24
major
compounds
were
isolated
and
fully
identified
using
spectroscopic
methods
comprising
flavonols,
flavan-‐3-‐ol
derivatives
and
phenolic
acids.
Among
isolated
compounds
five
malonylated
quercetin
and
kaempferol
derivatives
were
identified
(1-‐5,
Fig.
1).
Two
new
natural
products
namely
quercetin
3-‐O-‐(5”-‐O-‐malonyl-‐arabinofuranoside)
(2)
and
kaempferol
3-‐O-‐(5”-‐O-‐malonyl-‐arabinofuranoside)
(5)
were
isolated.
The
anti-‐
inflammatory
potential,
of
a
series
of
five
malonylated
flavonols
was
evaluated
using
human
neutrophils
model.
Compounds
were
tested
in
the
concentration
range
of
1-‐25
µM.
The
ability
to
inhibit
the
neutrophils
oxidative
burst
was
tested
and
the
influence
on
the
release
of
myeloperoxidase
(MPO)
was
also
checked.
Compounds
were
not
toxic
for
neutrophils
up
to
25
µM.
All
compounds
were
able
to
inhibit
the
production
of
reactive
oxygen
species
at
con-‐
centrations
of
1,
10
and
25
µM.
Compounds
1-‐3
were
stronger
inhibitor
than
compounds
4-‐5.
All
isolated
compound
also
inhibited
the
MPO
production,
but
only
compounds
1,
2
and
5
were
active
at
the
concentration
of
1
µM.
Compounds
1-‐3
at
concentrations
of
10
and
25
µM
were
stronger
inhibitors
than
the
positive
control
–
quercetin
(10
µM).
The
influence
of
com-‐
pounds
on
neutrophils
inflammatory
response
may
contribute
to
the
anti-‐inflammatory
po-‐
tential
of
P.
bistorta
as
a
food
product.
OH OH OH
OH OH OH
HO O HO O O O HO O
O
O OH O CH3
HOO O OH O
O O OH
OH O OH O OH OH O HO O
OH OH
O
O O
1 HO 2 3 O
OH OH
HO O HO O OH
O O
O
O OH O
O O O
HO OH O
OH O OH OH O OH
O OH
4 HO 5
Figure
1.
Chemical
structures
of
tested
rare
malonyl
flavonols
isolated
from
P.
bistorta.
Keywords:
Persicaria
bistorta,
flavonols,
neutrophils,
UHPLC-‐DAD-‐MS,
anti-‐inflammatory
References:
[1] Prance
G,
Nesbitt
M.
The
Cultural
History
of
Plants,
Taylor
&
Francis,
2012
[2] Kuźniewski
E,
Augustyn-‐Puziewicz
J.
Przewodnik
ziołolecznictwa
ludowego.
Warszawa-‐
Wrocław:
Polskie
Wydawnictwo
Naukowe,
1986
[3] Smolarz
HD.
Flavonoid
glycosides
in
nine
Polygonum
L.
taxons.
Acta
Soc
Bot
Pol
2002;
71:
29-‐33.
P451
Chemical
composition
of
Prangos
hulusii
Secil
Yazici-‐Tutunis,
Nur
Tan,
Mahmut
Miski
Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Istanbul
University,
34116
Istanbul,
Turkey
The
genus
Prangos
which
belongs
to
the
Apiaceae
family
is
widely
represented
in
the
world
and
is
commonly
known
as
“caksir”
in
Turkey
[1].
Members
of
this
genus
including
Prangos
hulusii
have
been
traditionally
used
as
a
wound/scar
healer,
emollient,
antifungal,
antihemor-‐
rhoidal,
antioxidant,
antihelmentic,
aphrodisiac,
antiflatulent
and
to
stop
bleeding
in
Turkey
and
Central
Asian
countries
[2−4].
Previous
chemical
investigations
on
the
secondary
metabo-‐
lites
of
Prangos
species’
have
resulted
in
the
isolation
of
various
coumarins,
alkaloids,
flavo-‐
noids
and
terpenoids
[5].
Only
limited
bioactivity
studies
have
been
performed
on
the
extracts
of
the
roots
and
aerial
parts
of
P.
hulusii
S.G.
Şenol,
H.
Yıldırım
&
Ö.
Seçmen
[6],
a
new
endemic
species
from
Flora
of
Turkey,
and
so
far
no
phytochemical
investigation
was
performed
on
this
species.
Dried
and
coarsely
powdered
roots
of
P.
hulusii
were
extracted
with
dichloro-‐
methane
in
a
Soxhlet
apparatus
and
the
solvent
was
evaporated
under
reduced
pressure
in
a
rotary
evaporator.
Four
known
coumarins
(oxypeucedanin,
murraol,
isoimperatorin
and
osthol),
two
steroids
(stigmasterol
and
β-‐sitosterol)
and
osthol-‐4’-‐senecioate,
a
new
couma-‐
rin,
were
isolated
and
identified
from
the
dichloromethane
extract
of
the
roots
of
P.
hulusii
using
various
chromatographic
and
spectroscopic
methods.
In
vivo
and
in
vitro
researches
have
showed
that
coumarins
have
many
pharmacological
properties
including
antispasmodic,
neuroprotective,
osteogenic,
immunomodulatory,
hepatoprotective,
cardiovascular
protective,
antiinflammatory
and
antimicrobial
activities
[7,
8].
Osthol
and
the
other
major
coumarins
present
in
P.
hulusii
may
be
responsible
for
the
aforementioned
therapeutic
effect
of
this
species.
Acknowledgements:
The
present
work
was
supported
by
the
Research
Fund
of
Istanbul
University.
Pro-‐
ject
No.
39751
Keywords: Prangos, coumarin, osthol-‐4'-‐senecioate
References:
[1] Baser KHC, Demirci B. Research into chemistry and biological activities of Prangos Lindl.
(Apiaceae) species of Turkey. Planta Med 2009; 75: SL19
[2] Razavi SM, Nazemiyeh H, Delazar A, Hajiboland R, Rahman M, Gibbons S, Nahar L, Sarkar
SD. Coumarins from the roots of Prangos uloptera. Phytochem Lett 2008; 1: 159–162
[3] Çoruh N, Sağdıçoğlucelep AG, Özgökçe F. Antioxidant properties of Prangos ferulacea L.,
Chaerophyllum macropodum Boiss. and Heracleum persicum Desf. from Apiaceae family
used as food in Eastern Anatolia and their inhibitory effects on Glutathione-‐stransferase.
Food Chem 2007; 100: 1237−1242
[4] Tada Y, Shikishima Y, Takaishi Y, Shibata H, Higuti T, Honda G, Ito M, Takeda Y,
Kodzhimatov OK, Ashurmetov O, Ohmoto Y. Coumarins and γ-‐pyrone derivatives from
Prangos pabularia: antibacterial activity and inhibition of cytokine release.
Phytochemistry 2002; 59: 649−654
[5] Shikishima Y, Takaishi Y, Honda G, Ito M, Takeda Y, Kodzhimatov OK, Ashurmetov O.
Terpenoids and γ-‐pyrone derivatives from Prangos tschimganica. Phytochemistry 2001;
57: 135−141
[6] Tütüniş-‐Yazıcı
S,
Tan
N,
Meriçli
F,
Özsoy
N,
Tan
E.
Biological
Activities
of
endemic
Prangos
hulusii.
Planta
Med
2013;
79:
PN113
[7] Zhang
ZR,
Leung
WN,
Cheung
HY,
Chan
CW.
Osthole:
A
review
on
its
bioactivities,
pharmacological
properties,
and
potential
as
alternative
medicine.
Evid-‐Based
Compl
Alt
2015;
2015:
919616
[8] Sadrei
H,
Shokoohinia
Y,
Sajjadi
SE
and
Ghadirian
B.
Antispasmodic
effect
of
osthole
and
Prangos
ferulacea
extract
on
rat
uterus
smooth
muscle
motility.
Res
Pharm
Sci
2012;
7:
141−149
P452
In
vitro
antidiabetic
activity
of
Cotinus
coggygria
Scop.
Sefa
Gözcü1,
Hafize
Yuca2,
Benan
Dursunoğlu2,
Zühal
Güvenalp2,
L.
Ömür
Demirezer3
1
Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Erzincan
University,
24100
Erzincan,
Turkey,
2
Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Atatürk
University,
25240
Erzurum,
Turkey,
3
De-‐
partment
of
Pharmacognosy,
Faculty
of
Pharmacy,
Hacettepe
University,
06100
Ankara,
Turkey
Cotinus
coggygria
Scop.
(Anacardiaceae)
is
a
deciduous
and
slow
growing
shrub.
It
grows
mainly
in
South
and
Central
Europe,
South
Russia,
Crimea,
Caucasia,
Latakia
and
Turkey
[1].
The
leaves
are
used
as
an
infusion
in
Turkish
folk
medicine
for
its
anti-‐diabetic,
antiseptic,
anti-‐inflammatory,
antimicrobial,
anti-‐haemorragic
and
wound
healing
properties
[2-‐3].
Ac-‐
cording
to
phytochemical
studies,
the
main
group
of
biologically
active
constituents
from
dif-‐
ferent
parts
of
C.
coggygria
are
polyphenols,
flavonoids
and
tannins.
The
plant
extracts
showed
antioxidant,
cytotoxic,
antigenotoxic,
antimicrobial,
antiviral,
hepatoprotective,
and
anti-‐inflammatory
activities
in
vivo
and
in
vitro.
There
are
also
a
few
studies
about
anti-‐
diabetic
activity
of
the
plant
[4].
In
the
present
study,
the
methanol
extract
and
its
different
polarity
fractions
(petroleum
ether,
dichloromethane,
ethyl
acetate,
n-‐butanol)
prepared
from
leaves
of
C.
coggygria
were
evaluated
for
in
vitro
α-‐glucosidase
and
α-‐amylase
inhibitory
ac-‐
tivities.
The
𝛼-‐glucosidase
of
Saccaharomyces
cerevisiae
is
used
to
investigate
the
inhibitory
activities
of
the
extracts
against
𝛼-‐glucosidase
by
using
𝑝-‐nitrophenyl-‐𝛼-‐D-‐glucopyranoside
as
a
substrate
and
these
were
compared
with
acarbose
[5].
The
dinitrosalicylic
acid
method
was
used
for
determination
of
α-‐amylase
inhibition
activity
and
expressed
as
a
decrease
in
units
of
maltose
released
[6].
The
ethyl
acetate
fraction
showed
the
best
α-‐glucosidase
inhibitory
ac-‐
tivity
with
96.06%
(IC50=0.0082
mg/mL)
when
compared
with
the
standard
compound
acar-‐
bose
that
displayed
2.17%
inhibitory
activity
(IC50=3.3642
mg/mL)
at
20
µg/mL
concentra-‐
tion.
The
petroleum
ether,
methanol,
n-‐butanol
and
dichloromethane
extracts
showed
15.45%
(IC50=0.0445
mg/mL),
13.87%
(IC50=0.0474
mg/mL),
11.60%
(IC50=0.0494
mg/mL)
and
5.81%
(IC50=0.1441
mg/mL)
inhibition
activity
at
the
same
concentration,
respectively.
How-‐
ever,
the
extracts
showed
no
inhibition
against
α-‐amylase.
Acknowledgements:
This
work
was
supported
by
The
Foundation
of
Atatürk
University
(2015/327).
Keywords:
Cotinus
coggygria
Scop.,
anti-‐diabetic
activity,
α-‐amylase
inhibition,
α-‐glucosidase
inhibition
References:
[1] Davis
PH,
Coode
MJE,
Cullen
J,
Cotinus
Adans.
In:
Davis
PH,
editors.
Flora
of
Turkey
and
The
East
Aegean
Islands.
Edinburgh
University
Press:
Edinburgh,
1982,
543.
[2] Baytop
T.
Türkiye'de
Bitkilerle
Tedavi
(Geçmişte
ve
Bugün).
İstanbul:
İstanbul
Üniversi-‐
tesi,
1999,
295.
[3] Arıtuluk
ZC,
Ezer
N.
Halk
arasında
diyabete
karşı
kullanılan
bitkiler
(Türkiye)-‐II.
Hacet-‐
tepe
University
J
Fac
Pharm
2012;
32:
179-‐208.
[4] Matić
S,
Stanić
S,
Mihailović
M,
Bogojević
D.
Cotinus
coggygria
Scop.:
An
overview
of
its
chemical
constituents,
pharmacological
and
toxicological
potential.
Saudi
J
Biol
Sci
2015;
http://dx.doi.org/10.1016/j.sjbs.2015.05.012
[5] Tao
Y,
Zhang
Y,
Cheng
Y,
Wang
Y.
Rapid
screening
and
identification
of
α-‐glucosidase
in-‐
hibitors
from
mulberry
leaves
using
enzyme-‐immobilized
magnetic
beads
coupled
with
HPLC/MS
and
NMR.
Biomed
Chromatogr
2013;
27:
148-‐155.
[6] Nampoothiri
SV,
Prathapan
A,
Cherian
OL,
Raghu
KG,
Venugopalan
VV,
Sundaresan
A.
In
vitro
antioxidant
and
inhibitory
potential
of
Terminalia
bellerica
and
Emblica
officinalis
fruits
against
LDL
oxidation
and
key
enzymes
linked
to
type
2
diabetes.
Food
Chem
Toxi-‐
col
2011;
49:
125-‐131.
P453
Neuroprotective effect of Perilla extracts on PC12 cells
Senavong Pimolvan1, Kongkham S2, Saelim S3, Suangkavathin V3
1 Department of Applied Thai Traditional Medicine, 2 Division of Biochemistry, Department of Preclinical
Sciences, 3 Research Center, Faculty of Medicine, Thammasat University, Phatumthani, 12120, Thailand.
Perilla
frutescens
(L.)
Britton
contains
rosmarinic
acid,
luteolin
and
linoleic
acid
which
had
neuroprotective
actions.
This
study
was
aimed
to
investigate
the
effect
of
Perilla
extracts
on
neuroprotection,
antioxidation
and
neurite
outgrowth
in
PC12
cells.
Freeze-‐dried
ethnolic
ex-‐
tract
from
Perilla
leaves
and
cold-‐pressed
oil
from
seed
at
dose
of
50,100,200
mg/ml
were
tested.
Cells
were
pretreated
with
each
extract
10
min
then
incubated
with
beta-‐amyloid
pro-‐
tein
20µM
for
24
h
to
induce
toxicity.
Cell
viability,
antioxidant
enzyme
(SOD)
activity
and
in-‐
hibition
on
tau-‐protein
hyperphosphorylation
were
analyzed
[1].
In
cell
cultured
in
serum
free
media
associated
with
2
ng/ml
of
nerve
growth
factor,
followed
by
adding
each
extract,
the
neurite
outgrowth
bearing
cells
and
MEK-‐1protein
production
were
investigated
[2].
Dulbecco’s
Modified
Eagle’s
Medium
and
rosmarinic
acid
were
used
as
normal
and
positve
controls
respectively.
Results
revealed
that
induction
of
toxicity
by
beta-‐amyloid
protein
in
PC12
cells:
(i)
the
cell
viability
decreased
to
60%
of
control
(ii)
the
cell
viability
increased
in
culture
pretreated
with
Perilla
leaf
extract
200
mg/ml
and
seed
oil
50
mg/ml;
(iii)
given
Peril-‐
la
leaf
extract
at
dose
of
100,
200
mg/ml
and
each
dose
of
seed
oil
to
cells
could
decrease
SOD
activity;
(iv)
tau
phosphorylation
at
Thr
205
and
Ser
396
was
decreased
by
pretreating
cells
with
Perilla
seed
oil
at
dose
of
50
mg/ml.
Moreover,
given
Perilla
leaf
extract
or
seed
oil
to
PC12
culture,
the
amount
of
neurite
outgrowth
bearing
cells
were
shown
to
increase
harmo-‐
niously
with
the
amount
of
MEK-‐1
protein.
In
summary,
Perilla
leaf
extract
and
seed
oil
re-‐
versed
the
effect
of
beta-‐amyloid
induced
toxicity
by
decreasing
oxidative
stress
as
well
as
inhibition
of
tau-‐protein
hyperphosphorylation.
The
enhancement
of
neurite
outgrowth
by
Perilla
extracts
was
also
revealed.
These
preliminary
results
imply
the
neuroprotective
effect
of
Perilla
as
a
functional
food.
Acknowledgements:
This
research
was
supported
by
grant
from
National
Research
Council
of
Thailand,
2014-‐2015.
Thanks
to
Professor
Dr.Maitree
Suttajit
for
providing
the
Perilla
extracts.
Keywords:
Perilla
frutescens,
neuroprotection.
References:
[1] Park
SY,
Kim
HS,
Cho
EK,
Kwon
BY,
Phark
S,
Hwang
KW,
et
al.
Curcumin
protected
PC12
cells
against
beta-‐amyloid-‐induced
toxicity
through
the
inhibition
of
oxidative
damage
and
tau
hyperphosphorylation.
Food
Chem
Toxicol
2008;
46:
2881-‐2887
[2] Yamazaki
M,
Chiba
K,
Mohri
T.
Fundamental
role
of
nitric
oxide
in
neuritogenesis
of
PC12h
cells.
Br
J
Pharmacol
2005;
146:
662-‐669
P454
Four
new
neolignans
from
the
fruits
of
Juglans
mandshurica
Max-‐
im.
SeonJu Park, Nanyoung Kim, Guijae Yoo, Jun Hyung Park, Seung Hyun Kim
College
of
Pharmacy,
Yonsei
Institute
of
Pharmaceutical
Science,
Yonsei
University,
85
Songdogwahak-‐ro,
Yeonsu-‐gu,
Incheon
406-‐840,
Korea
Juglans
mandshurica
Maxim.
(Juglandaceae),
commonly
known
as
the
Manchurian
walnut,
is
a
traditional
folk
medicine
used
for
treatment
of
dermatosis
and
to
relieve
aches
in
Korea
and
China
[1].
Compared
to
the
phytochemical
investigations
performed
on
its
root
and
the
stem
bark,
very
few
studies
have
been
done
on
chemical
constituent
study
of
the
fruits
of
J.
mands-‐
hurica.
In
this
study,
four
new
neolignans
along
with
six
known
neolignans
were
isolated
from
the
fruit
of
J.
mandshurica.
The
structures
of
four
new
compounds
were
determined
to
be
7S,8S-‐threo-‐4,7,9,9´-‐tetrahydroxy-‐3´-‐methoxy-‐8-‐O-‐4´-‐neolignan
(1),
7R,8S-‐erythro-‐4,7,9,9´-‐
tetrahydroxy-‐3´-‐methoxy-‐8-‐O-‐4´-‐neolignan
(2),
7S,8S-‐threo-‐4,7,9-‐trihydroxy-‐1´-‐
acrylaldehyde-‐8-‐O-‐4´-‐neolignan
(3)
and
7R,8S-‐erythro-‐4,7,9-‐trihydroxy-‐1´-‐acrylaldehyde-‐8-‐O-‐
4´-‐neolignan
(4)
on
the
basis
of
extensive
spectroscopic
methods
including
1D
and
2D
NMR
(HSQC,
HMBC,
COSY
and
ROESY)
and
CD
spectra
data.
This
is
the
first
report
on
the
isolation
of
neolignans
from
J.
mandshurica.
Acknowledgements:
This
research
was
supported
by
Basic
Science
Research
Program
through
the
Na-‐
tional
Research
Foundation
of
Korea
(NRF)
funded
by
the
Ministry
of
Education,
Science
and
Technology
(NRF-‐2011-‐0025129)
and
by
BrainKorea21PLUS
program
funded
by
National
Research
Foundation
of
Korea.
Keywords: Juglans mandshurica Maxim., Juglandaceae, neolignans
References:
[1] Liu
L,
Li
W,
Koike
K,
Zhang
S,
Nikaido
T.
New
α-‐tetralonyl
glucosides
from
the
fruit
of
Juglans
mandshurica.
Chem
Pharm
Bull
2004;
52:
566-‐569
P455
Heterocyclic
compounds
from
the
fruits
of
Morus
alba
L.
and
their
anti-‐angiogenesis
activities
Seoung
Rak
Lee1,
Seulah
Lee1,
Hee
Jeong
Eom1,
Hee
Rae
Kang1,
Jae
Sik
Yu1,
Tae
Kyoung
Lee1,
Jiwon
Baek1,
Dahae
Lee1,
Won
Se
Suh1,
Ki
Hyun
Kim1
1School
of
Pharmacy,
Sungkyunkwan
University,
Suwon
440-‐746,
Republic
of
Korea
The
mulberry
tree
(Morus
alba
L.)
is
mostly
found
in
semitropical
region
and
has
been
har-‐
vested
in
East-‐Asia,
especially
in
Korea.
The
fruits
of
M.
alba
(Moraceae)
have
been
used
for
a
variety
of
illnesses,
especially
to
treat
arthritis,
rheumatism
and
diabetes
in
Korean
traditional
medicines
[1].
Previous
phytochemical
investigations
on
the
fruits
of
M.
alba
reported
the
present
of
a
number
of
lignans,
flavonoids,
benzofuran
derivatives,
and
phenolic
constituents
[2-‐5].
Some
of
the
compounds
showed
a
variety
of
biological
activities
including
antioxidant,
cytotoxicity,
anti-‐inflammatory,
and
antibiotic
effects[2,3].
However,
there
has
been
rarely
reported
on
bioactive
heterocyclic
compounds
from
the
fruits
of
M.
alba.
In
an
effort
to
search
for
the
bioactive
compounds
from
Korean
indigenous
plants,
we
investigated
a
MeOH
extract
of
the
fruits
of
M.
alba,
which
led
to
the
isolation
of
one
new
oxolane
derivative
(1)
together
with
five
known
compounds
(2-‐6).
The
structures
of
isolated
compounds
were
identified
by
spectroscopic
methods
including
1H
NMR,
13C
NMR,
2D
NMR,
and
MS
analysis.
To
the
best
our
knowledge,
compound
6
is
reported
for
the
first
time
as
a
natural
product
in
this
study,
though
it
has
been
reported
as
a
synthetic
product.
In
addition,
all
isolated
compounds
were
examined
for
their
inhibitory
effects
on
angiogenesis
in
HUVECs.
The
action
mechanism
of
compound
3
was
identified
on
the
basis
of
western
blot
results.
OH O
O H H
R N
H O
HOOC O
HO O N
O H
4 R = OH 6
1 2 3
5 R = CO2CH3
Acknowledgements:
This
research
was
supported
by
Basic
Science
Research
Program
through
the
Na-‐
tional
Research
Foundation
of
Korea
(NRF)
funded
by
the
Ministry
of
Science,
ICT
&
Future
Planning
(2015R1C1A1A02037383)
Keywords: Morus alba, Moraceae, Heterocyclic compounds, Anti-‐angiogenesis effect
References:
[1] Yimam
M,
Lee
YC,
Kim
TW,
Moore
B,
Jiao
P,
Hong
M,
Kim
HJ,
Nam
JB,
Kim
MR,
Oh
JS,
Cleve-‐
land
S,
Hyun
EJ,
Chu
M,
Jia
Q.
Analgesic
and
anti-‐inflammatory
effect
of
UP3005,
a
botani-‐
cal
composition
containing
two
standardized
extracts
of
Uncaria
gambir
and
Morus
alba.
Pharmacognosy
Res
2015;
7:
S39-‐46
[2] Zelova
H,
Hanakova
Z,
Cermakova
Z,
Smeikal
K,
Dall
Acqua
S,
Babula
P,
Cvacka
J,
Hosek
J.
Evaluation
of
anti-‐inflammatory
activity
of
prenylated
substances
isolated
from
Morus
al-‐
ba
and
Morus
nigra.
J
Nat
Prod
2014;
77:
1297-‐1303
[3] Seo
KH,
Lee
DY,
Jeong
RH,
Lee
DS,
Kim
YE,
Hong
EK,
Kim
YC,
Baek
NI.
Neuroprotective
ef-‐
fect
of
prenylated
arylbenzofuran
and
flavonoids
from
Morus
alba
fruits
on
glutamate-‐
induced
oxidative
injury
in
HT22
hippocampal
cells.
J
Med
Food
2015;
18:
403-‐408
[4] Lee
WJ,
Choi
SW.
Quantitative
changes
of
polyphenolic
compounds
in
Mulberry
(Morus
alba
L.)
leaves
in
relation
to
varieties,
harvest
period,
and
heat
processing.
Prev
Nutr
Food
Sci
2012;
17:
280-‐285
[5] Choi
SJ,
Jeon
H,
Lee
CU,
Yoon
SH,
Bae
SK,
Chin
YW,
Yoon
KD.
Isolation
and
development
of
quantification
method
for
cyanidin-‐3-‐glucoside
and
cyanidin-‐3-‐rutinoside
in
Mulberry
fruit
by
high-‐performance
countercurrent
chromatography
and
high-‐performance
liquid
chromatography.
Nat
Prod
Sci
2015;
21:
25-‐29
P456
Essential
oils
from
Artemisia
copa
(Asteraceae)
and
Aloysia
deser-‐
ticola,
two
medicinal
species
used
in
the
Chilean
Atacama’s
com-‐
munity
(Antofagasta)
S.
Ortiz1,
Pierre
Champy2,
S.
Boutefnouchet1
1Laboratoire
de
Pharmacognosie,
UMR
CNRS
8638
COMETE,
Faculté
de
Pharmacie
de
Paris,Université
Paris-‐Descartes,
Sorbonne
Paris
Cité,
4,
av.
de
l’Observatoire,
75006
Paris.
France
;
2
Laboratoire
de
Pharmacognosie,
UMR
8076
CNRS
BioCIS,
Faculté
de
Pharmacie,
Université
Paris-‐Sud,
5
rue
J.-‐B.
Clément,
92296
Châtenay-‐Malabry,
France.
The
Chilean
altiplano
from
Arica
to
Atacama
(Altitude
of
2500
to
5000
m)
is
a
high
plain,
with
cool
temperatures,
semi-‐arid
and
arid
climate
and
an
elevated
level
of
UV
radiation,
where
we
find
the
Aymara
and
Atacama
native
people.
Atacama’s
people
traditional
medicine
is
rich,
and
medicinal
uses
are
described
for
more
than
200
plants.
Among
these
species,
two
repre-‐
sent
a
great
range
of
interest
without
serious
phytochemical
analysis
of
their
bioactive
com-‐
pounds:
Artemisia
copa
Phil.
var.
copa
(Asteraceae)
and
Aloysia
deserticola
(Phil.)
Lu-‐Irving
&
O’Leary
ex
Acantholippia
deserticola
(Verbenaceae).
A.
copa
var.
copa
is
used
for
digestive
troubles
and
headaches,
whereas
A.
deserticola
is
used
for
digestive
troubles,
breast
milk
pro-‐
duction,
urinary
infections,
colds
and
bronchitis
[1].
Plants
were
collected
with
a
representative
of
the
Taira
Community
(Antofagasta
Region)
at
five
locations
at
different
altitudes:
A.
copa
var.
copa
(at
3100
m)
and
A.
deserticola
(at
2800,
3000,
3200
and
3300
m).
Essential
oils
(EO)
obtained
by
hydro-‐distillation
were
analysed
by
GC-‐MS
and
GC-‐FID.
The
first
results
obtained
for
EO
analysis
showed
a
clearly
different
phyto-‐
chemical
composition
for
our
A.
copa
var.
copa
sample
than
the
literature
data
reported
for
a
sample
collected
in
the
Cochambamba
area
in
Bolivia.
The
major
constituent
(55%)
in
our
Chilean
sample
is
chrysanthenone
whereas
it
is
β-‐thujone
in
the
Bolivian
sample.
Chrysan-‐
thenone
may
produce
two
artifacts
formed
during
hydrodistillation
(filifolone
and
(Z)-‐
isogeranic)
[2]
but
they
are
not
detected
by
GC-‐MS
in
our
case.
In
the
case
of
A.
deserticola,
Bicyclic
monoterpenes
[3.1.0]
represent
the
major
constituents
(90
%):
sabinene,
α
and
β
thu-‐
jone
isomers.
Interestingly,
the
EO
composition
varied
with
altitude.
Variations
of
the
per-‐
centage
of
α-‐
and
β-‐thujone
decreased
(81
to
74%
and
11
to
0,5%
respectively)
whereas
sab-‐
inene
increased
(6
to
16%)
with
increasing
altitude.
Acknowledgements:
The
authors
thank
to
Félix
Galleguillos
for
helping
during
the
collection
of
plants
and
to
CONICYT
Scholarship
for
PhD
studies
support
of
S.
Ortiz.
Keywords: Ethnopharmacology, Artemisia copa, Aloysia deserticola, essential oils
References:
[1] Villagrán
C.,
Castro
V.
Ciencia
indígena
de
los
Andes
del
norte
de
Chile,
Editorial
Universi-‐
taria
S
A
2012;
174:
292.
[2] Lopez
Arce
JB,
Collin
G,
Garneau
F-‐X,
Jean
F-‐I,
Gagnon
H.
Essential
Oils
from
Bolivia.
III.
Asteraceae:
Artemisia
copa
Philippi.
J
Essent
Oil
Res
2004;
16:
554−557
P457
Determination
of
phenolic
compounds
of
Tribulus
terrestris
(Zy-‐
gophyllaceae)
Sevim
Küçük1,
Fatih
Göger2,
Neşe
Kırımer2
1
Department
of
Pharmaceutical
Botany,
Anadolu
University,
Faculty
of
Pharmacy,
26470,
Eskişehir,
Tur-‐
key;
2
Department
of
Pharmacognosy,
Anadolu
University,
Faculty
of
Pharmacy,
26470,
Eskişehir,
Turkey
The
genus
Tribulus
L.
(Zygophyllaceae)
known
as
“Çoban
Çökerten,
Demirdikeni”
is
repre-‐
sented
by
one
species
in
Anatolia
[1].
This
genus
has
well
known
biological
effects,
such
as
increasing
on
testosterone
level,
decreasing
intestine
weight,
protection
of
mercury
toxicity,
sex
reversal
in
fish,
protecting
oxidative
stress,
antiurolithiatic
and
antimicrobial
effects.
Its
biological
effects
is
generally
attributed
to
saponins,
flavonoids,
alkaloids,
glycosides
and
phy-‐
tosteroids
contents
[2,3].
In
the
present
study,
Tribulus
terrestris
L.
was
collected
from
Eskisehir,
on
August
2014.
Powdered
dried
aerial
parts
and
fruits
were
macerated
with
70
%
methanol.
The
extracts
were
analysed
by
LC-‐MS/MS
for
their
content
of
phenolic
compounds.
Quercetin
3-‐O-‐rutinoside
(rutin)
and
quercetin
dihexoside
was
determined
to
be
the
major
compounds
in
the
extracts.
Rutin
was
quantified
as
7.62
mg/g
extract
of
aerial
parts
whereas
its
content
in
fruits
was
determined
to
be
4.55
mg/
g
extract.
The
content
of
quercetin
dihexo-‐
side
was
determined
to
be
7.03
mg/g
herba
extract
and
4.34
mg/g
in
the
fruit
extract
(calcula-‐
tion
based
on
rutin).
Keywords:
Tribulus
terrestris,
Phenolic
compounds,
LC-‐MS/MS
References:
[1] Güner
A,
Aslan
S,
Ekim
T,
Vural
M,
Babaç
MT.
List
of
Turkish
Flora
(Vascular
Plants).
Publi-‐
cation
of
Nezahat
Gökyiğit
Botanical
Garden
and
Flora
Research
Foundation
2012;
888
[2] Şahin
A,
Duru
M.
Demir
dikeni
bitkisinin
biyokimyasal
ve
fizyolojik
etkileri.
VII.
Zootekni
Congress,
2005;
[3] Roaiah
MF,
El
Khayat
YI,
GamalEl
Din
SF,
Abd
El
Salam
MA.
Pilot
Study
on
the
Effect
of
Bo-‐
tanical
Medicine
(Tribulus
terrestris)
on
Serum
Testosterone
Level
and
Erectile
Function
in
Aging
Males
With
Partial
Androgen
Deficiency
(PADAM).
J
Sex
Marital
Ther
2005;
7:
1-‐
5
P458
Promising Neuroprotective and Anti-‐Inflammatory Effect of Small
Nettle’ Ethyl Acetate Extract
Sevki Arslan1, Gulsum Terzioglu1, Buket Kabalay1, Ali Riza Tufekci2, Alaattin Sen1, Ibrahim
Demirtas2,
1 Pamukkale University, Faculty of Arts & Sciences, Department of Biology, Denizli, TURKEY; 2 Çankırı
Karatekin
University,
Faculty
of
Arts
&
Sciences,
Department
of
Chemistry,
Turkey
Multiple
sclerosis
(MS)
is
one
of
the
most
common
inflammatory
diseases
of
the
central
nerv-‐
ous
system.
Release
of
pro-‐inflammatory
mediators
initiates
the
cascades
of
events
leading
into
development
of
this
neurodegenerative
disease.
Due
to
its
progressive
neurodegenera-‐
tive
nature,
treatments
that
show
direct
neuroprotective
effects
are
very
promising
for
the
MS.
Recently,
plant
based
products
may
be
used
as
an
alternative
treatment
of
this
disease.
In
this
regard,
this
study
was
aimed
to
investigate
potential
neuroprotective
and
anti-‐
inflammatory
effects
of
Urtica
Urens’
(Small
nettle)
leaves
ethyl
acetate
extract
in
Caco2
cell
line.
In
this
respect,
firstly
extracts
of
dried
stinging
nettle
leaves
were
prepared
and
chemical
composition
of
extract
was
determined
by
LC-‐MS.
We
analysed
that
extract
increased
the
ex-‐
pression
of
nuclear
factor
(erythroid-‐derived
2)-‐like
2,
(Nrf2)
at
both
levels
of
RNA
and
pro-‐
tein
(80%
and
67%,
respectively).
It
is
well
established
that
this
transcription
factor
is
in-‐
volved
in
protection
against
oxidative
damage
triggered
by
injury
and
inflammation.
Extract
treatment
caused
15%
and
20%
decreases
in
protein
and
mRNA
levels
of
TNF-‐α,
respectively.
On
the
other
hand,
NF-‐κB
protein
and
mRNA
levels
increased
due
to
extract
treatment
(56%
and
48%,
respectively).
Moreover,
whole
genome
transcriptome
analysis
was
performed
by
using
Human
HT-‐12
V4
Bead
Chip
with
over
47.000
probes.
Total
275
different
probes
were
significantly
changed
(p<0.05)
by
ethyl
acetate
extract.
Transcriptome
profiling
analysis
showed
that
several
genes
involved
in
antigen
receptor-‐mediated
signalling
pathway,
immune
response-‐regulating
signal
transduction,
T
cell
receptor
signalling
pathway,
leukocyte
lym-‐
phocyte
and
T
cell
activation,
immune
system
development
were
modulated.
In
conclusion,
all
of
these
results
put
the
hypothesis
that
the
small
nettle’s
ethyl
acetate
extract
contains
prom-‐
ising
phytochemicals
that
may
be
used
in
neurodegenerative
diseases
treatment.
Acknowledgements: This work is supported by TUBITAK 111T515.
Keywords: Urtica urens, neuroprotection, anti-‐inflamatory activity
P459
Antiviral
and
antibacterial
effects
of
16
plants
used
in
Ivorian
traditional
medicine
Moussa
Bamba1,2,
Simon
Bordage1,
Marie-‐Emmanuelle
Sahuc3,
Jennifer
Samaillie1,
Christel
Neut4,
Alexis
Zamble5,
Fezan
Tra
Bi2
,
Karin
Seron3,
Sevser
Sahpaz1
1
Univ.
Lille,
INRA,
ISA,
Univ.
Artois,
Univ.
Littoral
Côte
d’Opale
-‐
EA
7394
-‐
Institut
Charles
Viollette,
F-‐
59000
Lille,
France,
2
Université
Nangui
Abrogoua,
02
BP
801
Abidjan
02,
Ivory
Coast,
3
Infection
and
Immunity
Centre,
CNRS
UMR8204-‐INSERM
U1019,
59000
Lille,
France,
4
Université
de
Lille
-‐
Labora-‐
toire
de
Bactériologie,
BP
83,
F-‐59000
Lille,
France,
5
Université
Lorougnon
Guédé,
BP
150
Daloa,
Côte
d’Ivoire
Over
160
million
people
are
chronic
carriers
of
Hepatitis
C
Virus
(HCV),
which
is
a
major
cause
of
chronic
liver
diseases1.
A
few
efficient
antiviral
drugs
against
HCV
are
currently
available,
but
they
are
expensive
and
not
easily
accessible
to
people
in
developing
countries2.
In
addition,
resistance
to
these
new
drugs
are
already
emerging.
Therefore
new
HCV
inhibi-‐
tors
are
needed.
A
major
symptom
of
HCV
infection
is
icterus
and
could
correspond
to
the
“yellow
malaria”
known
in
African
traditional
medicines.
An
ethnobotanical
survey
and
a
bib-‐
liographic
search
allowed
us
to
select
16
Ivorian
plants,
based
on
their
traditional
use
against
“yellow
malaria”
and
other
infectious
diseases.
For
each
plant,
20
g
of
leaf,
root
or
bark
powder
were
macerated
in
100
mL
of
methanol
(3
x
24
h).
The
resulting
extracts
were
then
screened
against
36
strains
of
bacteria
using
an
agar
dilution
method
with
a
Steers
multipoint
inoculator,
and
against
HCV
using
an
immunofluo-‐
rescence
method
with
Huh-‐7
cells
(extracts
at
10
or
25
µg/mL).
Ten
extracts
were
active
on
at
least
one
strain
of
bacteria
at
0.6
mg/mL
or
less,
with
one
ex-‐
tract
(Anogeissus
leocarpus)
(Combretaceae)
being
active
on
19
strains.
The
Momordica
char-‐
antia
(Cucurbitaceae)
extract
was
particularly
active
on
3
Streptococcus
strain,
with
some
MIC
as
low
as
40
µg/mL.
Carapa
procera
(Meliaceae)
and
Pericopsis
laxiflora
(Fabaceae)
showed
very
interesting
activities
against
HCV.
For
these
2
plants,
leaf
powder
(20
g)
was
successively
macerated
in
100
mL
of
dichloromethane,
methanol
and
ethanol-‐water
(50:50)
respectively
(3
x
24
h
extraction
per
solvent).
The
resulting
6
dried
extracts
were
then
dissolved
in
metha-‐
nol
and
screened
against
HCV.
Amongst
them,
the
dichloromethane
were
not
active,
the
hy-‐
dro-‐alcoholic
extracts
were
mildly
active
(>50%
infected
cells),
but
the
methanolic
extracts
(10
µl/mL)
prevented
over
80%
of
Huh-‐7
cells
from
being
infected.
Bio-‐guided
fractionations
of
several
plant
extracts
are
in
progress.
Keywords:
Ivorian
pharmacopeia,
ethnobotanical
survey,
bio-‐guided
fractionation,
antibacte-‐
rial,
antiviral,
HCV
References:
[1] Dhingra
A.,
Kapoor
S,
Alqahtani
SA.
Recent
advances
in
the
treatment
of
hepatitis
C.
Discov
Med
2014;
18:
203-‐208
[2] Calland
N,
Dubuisson
J,
Rouillé
Y,
Séron
K.
Hepatitis
C
virus
and
natural
compounds:
a
new
antiviral
approach?
Viruses
2012;
10:
2197-‐2217
P460
Antimicrobial
and
acetylcholinesterase
inhibition
activities
of
an
endemic
plant
from
Sahara:
Myrtus
nivellei
Batt.
&
Trab.
(Myr-‐
taceae)
Rym
G.
Demmak1,2,
Simon
Bordage2,
Christel
Neut3,
Abderahman
Bensegueni1,
Sevser
Sahpaz2
1
Department
of
natural
science
and
life,
University
of
Constantine
1,
25000,
Constantine,
Algeria,
2
Univ.
Lille,
INRA,
ISA,
Univ.
Artois,
Univ.
Littoral
Côte
d’Opale
-‐
EA
7394
-‐
Institut
Charles
Viollette,
F-‐59000
Lille,
France,
3Université
de
Lille
-‐
Laboratoire
de
Bactériologie,
BP
83,
F-‐59000
Lille,
France
Myrtus
nivellei
is
a
medicinal
plant
used
traditionally
in
Algeria
to
treat
several
disorders
in-‐
cluding
skin
diseases,
diarrhea,
fever
and
diabetes1.
Very
little
is
known
about
the
phytochem-‐
istry
and
biological
activities
of
this
plant,
which
might
contain
antimicrobial
compounds.
In
addition,
its
close
relative
M.
communis
is
well
studied
and
contains
inhibitors
of
acetylcholin-‐
esterase
(AChE),
an
enzyme
that
is
currently
targeted
to
treat
symptoms
of
Alzheimer’s
dis-‐
ease.
The
aim
of
this
study
is
to
investigate
antimicrobial
and
anti-‐AChE
activities
in
M.
nivellei
and
to
purify
new
active
compounds.
A
crude
methanol/water
(80:20)
extract
of
M.
nivellei
aerial
parts
(500
g)
was
fractionated
by
liquid/liquid
partition
using
three
solvents
of
increas-‐
ing
polarity.
From
500
g
of
dried
plant
material
we
obtained
71.4
g,
1.2
g,
21.4
g,
2.1
and
31.4
g
of
hydro-‐alcoholic,
petroleum
ether,
CHCl3,
EtOAc
and
n-‐BuOH
dried
extracts,
respectively.
All
extracts
were
then
tested
by
bioautography
for
their
anti-‐AChE
activity2.
The
bioautog-‐
raphy
revealed
that
the
crude
extract
and
the
the
chloroformic
fraction
contain
at
least
five
active
compounds
with
AChE
inhibitory
activity.
The
chloroformic
extract
(19
g)
was
then
further
fractionated
with
an
open
column
chromatography
using
different
solvents
in
various
proportions.
24
sub-‐fractions
were
obtained
and
tested
for
their
inhibitory
activity
on
AChE.
Several
of
them
were
still
active,
including
the
5
major
components
but
also
other
molecules.
The
most
active
components
will
be
purified
and
further
studied
with
AChE
microplate
as-‐
says.
In
addition,
the
crude
hydro-‐alcoholic
extract
was
tested
on
Hepatitis
C
Virus
(HCV)
and
36
strains
of
bacteria.
Although
it
showed
no
antiviral
activity,
the
crude
extract
was
active
(at
0.6
mg/mL
or
less)
on
7
strains
of
bacteria,
including
Staphylococcus
epidermidis
and
Myco-‐
bacterium
smegmatis,
which
can
be
involved
in
skin
diseases.
Further
studies
are
in
progress
to
identify
new
active
compounds.
Keywords:
Myrtus
nivellei,
acetylcholinesterase
inhibition,
antimicrobial
assays,
bio-‐guided
fractionation,
bioautography.
References:
[1] Bouzabata
A,
Bazzali
O,
Cabral
C,
Gonçalves
MJ,
Cruz
MT,
Bighelli
A,
Cavaleiro
C,Casanova
J,
Salgueiro
L,
Tomi
F.
New
compounds,
chemical
composition,
antifungal
activity
and
cyto-‐
toxicity
of
the
essential
oil
from
Myrtus
nivellei
Batt.
&
Trab.,
an
endemic
species
of
Cen-‐
tral
Sahara.
J
Ethnopharmacol
2013;
149:
613-‐620.
[2] Yang
Z,
Zhang
X,
Duan
D,
Song
Z,
Yang
M.
Modified
TLC
bioautographic
method
for
screen-‐
ing
acetylcholinesterase
inhibitors
from
plant
extracts.
J
Sep
Sci
2009;
32:
3257-‐3259.
P461
Effect
of
Crocus
sativus,
as
an
accompaniment
agent
in
traditional
Persian
medicine,
on
pharmacokinetic
of
acetaminophen
Seyede
Nargess
Sadati1,
Sima
Sadrai2,
Mohammad
R.
Shams
Ardekani3,
Katayoon
Mireskandari
2,
Mohammad
Sharifzadeh4
1
Department
of
Traditional
Pharmacy,
School
of
Traditional
Iranian
Medicine,
Tehran
University
of
Med-‐
ical
Sciences,
Tehran,
Iran;
2
Department
of
Pharmaceutics,
Faculty
of
Pharmacy,
Tehran
University
of
Medical
Sciences,
Tehran,
Iran;
3
Department
of
Pharmacognosy
and
Persian
Medicine
and
Pharmacy
Research
Center,
Faculty
of
Pharmacy,
Tehran
University
of
Medical
Sciences;
4
Department
of
Pharma-‐
cology
and
Toxicology,
Faculty
of
Pharmacy,
Tehran
University
of
Medical
Sciences,
Tehran,
Iran
One
concept
used
in
traditional
Persian
Medicine
for
multi-‐drug
therapy
is
the
use
of
accom-‐
paniment
agents
(Mobadregh).
According
to
TPM
texts,
accompaniments
are
substances
(or
drugs),
which
facilitate
access
to
the
whole
body
or
to
specific
organs
for
drugs
or
food.
[1,2]
This
study
investigated
the
effect
of
oral
co-‐administration
of
Crocus
sativus
(saffron)
on
the
absorption
and
some
pharmacokinetic
parameters
of
acetaminophen
in
rats.
To
the
test
group
only
acetaminophen
10
mg/kg
and
to
the
concomitant
group
acetamino-‐
phen
10
mg/kg
and
Crocus
sativus
4
mg/kg
(n=6
at
each
group)
were
administered.
The
plasma
acetaminophen
levels
(at
0,
5,
10,
15,
20,
40,
60,
90,
and
120
min
after
oral
administra-‐
tion)
were
monitored
by
an
HPLC-‐UV
method.
Results
indicated
that
co-‐administration
of
ac-‐
etaminophen
and
Crocus
sativus
significantly
increased
acetaminophen’s
area
under
concen-‐
tration
curve
(AUC)
compared
to
acetaminophen
alone.
The
AUC0-‐90
of
the
test
group
was
333.5±136
min_µg/mL
and
the
AUC0-‐90
of
the
concomitant
group
was
414.7±194
min_µg/mL.
These
results
suggested
that
saffron,
one
of
the
accompaniment
agents
in
TPM,
could
increase
bioavailability
of
acetaminophen
and
can
be
considered
as
an
enhancer
of
bio-‐
availability
and
efficacy
of
other
drugs
at
least
by
oral
route
and
also
the
drug
interactions
with
this
herbal
drug
should
be
considered.
Keywords:
acetaminophen,
accompaniment
agent,
Crocus
sativus,
pharmacokinetic,
tradi-‐
tional
Persian
medicine
References:
[1]
Ebadi
N,
Masoomi
F,
Yakhchali
M,
Sadati
Lamardi
SN,
Shams-‐Ardakani
MR,
Sadeghpour
O,
Raiesdana
A,
Ramezany
F.
Convoy
drugs
in
traditional
Persian
medicine:
The
historical
concepts
of
bioavailability
and
targeting.
Trad
Integr
Med
2015;
1:
18-‐27
[2]
Sadati
SN,
Ardekani
MR,
Ebadi
N,
Yakhchali
M,
Dana
AR,
Masoomi
F,
Khanavi
M,
Ramezany
F.
Review
of
scientific
evidence
of
medicinal
convoy
plants
in
traditional
Persian
medi-‐
cine.
Pharmacogn
Rev
2016;
10:
33-‐38
P462
Quantitative
analysis
of
phenolic
compounds
of
some
Crataegus
species
from
Turkey
Sezin
Anil1,
Esra
Eroğlu
Özkan1,
Ali
Dönmez2,
Gülay
Melikoğlu1
1
Department
of
Pharmacognosy,
Istanbul
University,
Faculty
of
Pharmacy,
34116,
Istanbul,
Turkey,
2
Department
of
Biology,
Division
of
Botany,
Hacettepe
University,
Faculty
of
Science,
06800,
Ankara,
Tur-‐
key
Crataegus
(hawthorn)
species
are
used
as
folk
medicine
all
around
the
world.
Most
of
the
studies
with
Crataegus
species
focus
on
effects
on
heart
failure
and
cardiovascular
disease
[1].
The
extract
is
required
to
be
standardized
according
to
its
flavonoid
(vitexine-‐2″-‐O-‐
rhamnoside
or
hyperoside)
and
oligomeric
procyanidin
content
by
the
European
Pharmaco-‐
poeia
(EP)[2].
The
present
study
aimed
to
research
on
the
leaves
and
fruits
of
Crataegus
petrodavisii
Dönmez
and
C.
christensenii
Dönmez
growing
in
Turkey.
Their
phytochemical
content
were
evaluated
by
HPLC
using
chlorogenic
acid,
caffeic
acid,
vitexin,
vitexin
2”-‐O
rhamnoside,
vitexin
4”-‐O
glycoside,
rutin,
hyperoside,
isoquercetin,
quercitrin,
kaempferol
3-‐
glycoside,
kaempferol,
apigenin,
apigenin
7-‐glycoside,
spiraeoside,
luteolin
and
quercetin
standards.
HPLC
analysis
is
performed
on
a
ACE
5
C18
analytical
column
(250
×
4.6
mm,
i.d.,
5-‐µm)
using
solvent
(A)
acetonitrile-‐tetrahydrofuran
(95:5,
v/v)
and
(B)
1%
aqueous
phos-‐
phoric
acid
as
mobile
phase
with
UV
absorption
at
336
and
366
nm[3].
As
a
result,
vitexin
is
the
major
compound
for
C.
petrodavisii
leaves
(0,162
mg/ml)
and
C.
christensenii
leaves
(0,201
mg/ml).
While
rutin
is
the
major
compound
for
C.
petrodavisii
fruit
(0,024
mg/ml),
hyperoside
is
major
for
C.
christensenii
fruit
(0,017
mg/ml).
Acknowledgements:
The
present
work
was
supported
by
the
Research
Found
of
Istanbul
University.
Pro-‐
ject
No.
39214
Keywords: Crataegus, hawthorn, HPLC, phenolic compounds
References:
[1] ESCOP
Monographs,
the
European
Scientific
Cooperative
on
Phytotherapy
2nd
editions.
New
York:
Thieme;
2003:
98-‐106
[2] European
Pharmacopoeia
5.0.
Strasburg
:
The
Council
of
Europe;
2004:1712-‐1715
[3] Ying,
X.,
Wang,
R.,
Xu,
J.,
Zhang,
W.,
Li,
H.,
Zhang,
C.
and
Li,
F.
HPLC
determination
of
eight
polyphenols
in
the
leaves
of
Crataegus
pinnatifida
Bge.
var.
major.
J.
Chromatogr.
Sci.
2009;
47,
3:
201–205
P463
Cytoprotective
and
antiadhesive
effects
of
aqueous
leaf
extract
from
Orthosiphon
aristatus
against
uropathogenic
E.
coli
Shabnam
Sarshar1,
Mohammad
R.
Asadi
Karam3,
Mehri
Habibi3,
Saeid
Bouzari3,
Xiaofei
Qin2,
Francisco
M.
Goycoolea2,
Simone
Brandt1,
Andreas
Hensel1
¹
University
of
Münster,
Institute
of
Pharmaceutical
Biology
and
Phytochemistry,
Corrensstraße
48,
D-‐
48149
Münster,
Germany,
2
University
of
Münster,
Institute
of
Biology
and
Biotechnology
of
Plants,
Schlossgarten
3,
D-‐48149
Münster,
Germany,
3
Department
of
Molecular
Biology,
Pasteur
Institute
of
Iran,
Pasteur
Ave.,
Tehran
13164,
Iran
Uropathogenic
E.
coli
(UPEC)
adherence
to
host
cells
is
a
crucial
step
for
infection
onset
in
the
urogenital
tract.
Targeting
UPEC
adhesion
becomes
an
alternative
approach
to
prevent
UTIs.
This
study
investigated
the
antiadhesive
potential
of
an
aqueous
extract
from
the
leaves
of
Orthosiphon
aristatus
(Blume)
Miq
against
different
UPEC
strains
in
an
in
vivo
(a
mouse
infec-‐
tion
model)
and
in
vitro
(flow
cytometric
adhesion
assay).
Dried
plant
material
was
extracted
with
hot
water
(WE,
drug-‐extract
ratio
5:1)
and
characterized
by
LC-‐MS.
Within
the
in
vivo
experiment
mice
were
pretreated
for
4
and
7
days
with
WE
(500,
750
mg/kg;
Norfloxacin
as
positive
control).
Subsequently,
mice
were
inoculated
transurethrally
with
2
×
108
CFU
UPEC
strain
NU14.
24h
post
infection
bladder
tissue
was
homogenized
and
the
suspension
used
for
determination
of
bacterial
load.
Both,
4
and
7
days
pretreatment
significantly
reduced
the
bacterial
colonization
(Fig.
1).
WE
(750
mg/kg,
7
day
pretreatment)
reduced
also
bacterial
colonization
of
kidneys
after
infection
with
UPEC
strain
CFT073
in
a
similar
experiment.
To
confirm
the
in
vivo
activity,
WE
was
tested
within
in
vitro
assays
and
no
cytotoxic
effects
up
to
2
mg/mL
were
observed
against
UPEC
as
well
as
against
T24
bladder
cells.
Coincubation
of
UPEC,
T24
cells
and
WE
increased
the
bacterial
adhesion
due
to
cell
agglomeration,
as
shown
by
flow
cytometry
and
Giemsa
staining.
In
contrast
preincubation
of
UPEC
with
WE
in
pooled
urine
led
to
40%
reduction
of
bacterial
adhesion.
Interestingly
coincubation
of
tannin-‐free
extract
of
WE
(WEøTannin)
did
not
cause
any
bacterial
aggregation.
Preincubation
of
UPEC
with
WEøTannin
in
urine
reduced
the
bacterial
attachment
up
to
50%.
This
anti-‐adhesive
ef-‐
fect
of
WE
was
correlated
by
RT-‐PCR
to
the
changed
gene
expression
of
selected
virulence
factors
(significant
downregulation
of
FimH,
upregulation
of
FliC).
Additionally
significant
inhibition
of
bacterial
quorum
sensing
(AHL/LuxR
biosensor
assay)
was
detected.
Fig.
1:
Influence
of
pre-‐treatment
of
mice
with
WE
at
concentration
of
500
and
750
mg/kg
on
the
bacterial
colonization
in
mice
bladder
tissue
24
h
after
infection
with
UPEC
strain
NU14.
UC:
untreated
control.
Keywords:
Urinary
tract
infection,
UPEC,
Orthosiphon
aristatus,
anti-‐adhesion
effect,
Murine
UTI
model,
PCR,
AFM
P464
New
cytotoxic
constituents
from
Xestospongia
testudinaria
sponge
collected
from
the
Red
Sea
of
Saudi
Arabia
Ali
A.
El-‐Gamal1,2,
Shaza
M.
Al-‐Massarani1,
Diaa
T.
A.
Youssef3,
Lamiaa
A.
Shaala4,
Mansour
S.
Al-‐Said1,
Abdelkader
E.
Ashour5,
Ashok
Kumar6,
Maged
S.
Abdel-‐Kader7,
Wael
M.
Ab-‐
del-‐Mageed1
1
Department
of
Pharmacognosy,
College
of
Pharmacy,
King
Saud
University,
P.O.
Box
2457,
Riyadh
11451,
Saudi
Arabia,
2
Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Mansoura
University,
El-‐
Mansoura
35516,
Egypt,
3
Department
of
Natural
Products,
Faculty
of
Pharmacy,
King
Abdulaziz
Univer-‐
sity,
Jeddah
21589,
Kingdom
of
Saudi
Arabia,
4.Natural
Products
Unit,
King
Fahd
Medical
Research
Cen-‐
ter,
King
Abdulaziz
University,
Jeddah
21589,
Kingdom
of
Saudi
Arabia,
5
Department
of
Pharmacology
and
Toxicology,
College
of
Pharmacy,
King
Saud
University,
Riyadh
11451,
P.O.
Box
2457,
Saudi
Arabia,
6
Vitiligo
Research
Chair,
College
of
Medicine,
King
Saud
University,
Riyadh,
Saudi
Arabia,
7
Department
of
Pharmacognosy,
College
of
Pharmacy,
Sattam
Bin
Abdulaziz
University,
Al-‐kharj
11942,
Saudi
Arabia
Since
the
beginning
of
the
exploration
of
marine
natural
products
in
the
early
1970’s,
several
studies
on
the
secondary
metabolites
of
Xestospongia
spp
(family
Petrosiidae),
commonly
known
as
barrel
sponges,
have
been
published
from
several
regions
around
the
world
[1,2].
Bioassay
guided
fractionation
of
X.
testudinaria
sponge,
collected
from
the
eastern
coast
of
the
Red
sea,
Saudi
Arabia,
led
to
the
isolation
of
fourteen
compounds.
The
nucleoside
derivative,
5-‐(6-‐amino-‐3H-‐imidazo[4,5-‐β]
pyridine-‐3-‐yl)-‐2
(hydroxymethyl)
tetrahydrofuran-‐3-‐ol
(1),
and
two
xestosterol
esters
identified
as
xestosterol
palmitate
(2)
and
xestosterol
of
l6-‐bromo
(7E,11E,l5E)
hexadeca-‐7,11,l5-‐trien-‐5,13-‐diynoic
acid
(3)
have
been
isolated
for
the
first
time.
The
known
compounds
2-‐methyl
maleimide
5–oxime,
maleimide-‐5–oxime,
tetillapyrone,
nor-‐
tetillapyrone,
xestosterol,
xestosterol
ester
of
18-‐bromooctadeca-‐9E,7E-‐diene-‐7,15-‐diynoic,
five
brominated
acetylenic
fatty
acid
derivatives
were
also
isolated.
The
chemical
structures
were
determined
based
on
extensive
spectroscopic
experiments.
The
total
alcoholic
and
n-‐
hexane
extracts;
as
well
as
18,18-‐dibromo-‐9E-‐octadeca-‐9,17-‐dien-‐5,7-‐diynoic
acid
showed
remarkable
cytotoxic
activity
against
Hela,
HepG2
and
Daoy
cancerous
cell
lines
with
IC50
of
35.07,
33.7
and
23.85
µg/ml
respectively
compared
to
the
reference
drug
Dasatinib
(IC50
16.22
µg/ml).
The
other
isolated
compounds
demonstrated
lower
cytotoxic
activity
(IC50
30.38-‐44.41µg/ml).
N NH2
N N
O
O
HO
OH 1 O 2
14
15'
Br
13' 11' 7'
O
5'
O
3
Acknowledgements:
The
authors
are
thankful
to
the
deanship
of
Scientific
Research
at
King
Saud
Univer-‐
sity
for
funding
the
work
through
the
Research
Group
Project
no.
RGP-‐VPP-‐326.
Keywords: Xestospongia testudinaria, cytotoxic activity, nucleoside derivative, xestosterol
ester, brominated acetylenic fatty acids.
References:
[1] Zhou
X,
Xu
T,
Yang
XW,
Huang
R,
Yang
B,
Tang
L
Liu
Y.
Chemical
and
biological
aspects
of
marine
sponges
of
the
genus
Xestospongia.
Chem
Biodivers
2010;
7:
2201-‐2227.
[2] Brantley
SE,
Molinski
TF,
Preston
CM,
DeLong
EF.
Brominated
acetylenic
fatty
acids
from
Xestospongia
sp.,
a
marine
spongebacteria
association.
Tetrahedron
1995;
51:7667-‐7672.
P465
Screening
of
compounds
of
Evernia
prunastri
(L.)
for
their
anti-‐
proliferative
activity
in
glioblastoma
cells
Shcherbakova
A1,3,
Nyugen
L1,2,
Koptina
A2,
Backlund
A2,
Shurgin
A3,
Romanov
E3,
Ulrich-‐
Merzenich
G1
1
Medical
Clinic
III,
UKB,
Friedrich
Wilhelms
University
of
Bonn,
Sigmund-‐Freudstr.
25,
53127
Bonn
Uni-‐
versity,
Germany,
2
Division
of
Pharmacognosy,
Department
of
Medicinal
Chemistry,
Uppsala
University,
BMC-‐Biomedical
center,
Box
574,
S-‐751
23
Uppsala,
Sweden,
3
Department
of
Forestry,
Ploshchad'
Lenina,
3,
Yoshkar-‐Ola,
Mari
El
Republic,
424000,
Russian
Federation
Lichens
possess
a
broad
spectrum
of
biological
activity,
which
is
probably
related
to
exclu-‐
sively
produce
secondary
metabolites
from
the
lichenized
fungi.
Those
are
depsides,
depsido-‐
nes
and
usnic
acid
derivatives
[1].
Previously
we
showed
that
the
acetonitrile
extract
from
Evernia
prunastri
(L.)
is
cytotoxic
for
human
glioblastoma
cells
(U-‐87
cell
line)
(IC50:
29.3µM)
and
less
toxic
for
human
fibroblasts,
and
that
this
a
biological
activity
not
related
to
usnic
acid
[2].
Another
major
compound
of
Evernia
prunastri
is
evernic
acid
(EA).
Here
we
investigated
the
capability
of
EA
to
inhibit
the
proliferation
of
glioblastoma
cells
and
to
synergize
with
the
standard
medication
for
glioblastoma
-‐
temozolamide
(TMZ)
or
with
catechin
in
reference
to
the
proanthocyanidin
content
of
selected
lichens
[3]
and
to
Parmotrema
reticulatum
which
was
detected
to
induce
apoptosis
in
certain
cancer
cell-‐lines
[4].
The
viability
of
cells
was
in-‐
vestigated
by
the
resazurin
assay.
Combinatory
effects
were
calculated
by
the
Chou-‐Talalay
method
[5].
Pathways
involved
in
cell
proliferation
and
apoptosis
were
evaluated
by
Western
Blot.
EA
exhibited
an
IC50
of
62μM
against
U-‐87
cells,
whereas
catechin
and
TMZ
inhibited
50%
of
cell
proliferation
in
concentrations
of
719μM
and
593μM
respectively.
Furthermore,
it
was
concluded
that
EA
influenced
cell
proliferation
through
the
phosphorylation
of
MAP-‐
kinases,
and
that
EA
did
not
synergize
with
either
catechin
or
TMZ
(CI>1).
The
indicated
'mode
of
action'
on
MAP-‐kinases
is
consistent
with
evaluations
using
the
ChemGPS-‐NP
chemi-‐
cal
property
space
[6],
and
comparing
with
a
reference
set
of
well-‐defined
activities
as
exem-‐
plified
by
NCI
(see
figs.
1a-‐e).
Thus,
EA
may
contribute
to
the
anti-‐glioblastoma
activity
of
Evernia
prunastri
but
the
search
for
additional
compounds
or
compound
combinations
from
this
lichen
that
exhibit
high
anticancer
activity
and
an
ability
to
synergize
will
be
continued.
a. b. c.
d. e.
Figure
1a-‐e.
Position
in
chemical
property
space
of
evernic
acid
(orange
cube),
compared
to
reference
compounds
of:
a.
tubulin-‐active,
b.
antimetabolite,
c.
topoisomerase
I,
d.
topoiso-‐
merase
II,
and
e.
alkylator
types
Acknowledgements: A. Shcherbakova was supported by the DAAD
References
[1] Stocker-‐Worgotter
E.
Metabolic
diversity
of
lichen-‐forming
ascomycetous
fungi:
culturing,
polyketide
and
shikimate
metabolite
production,
and
PKS
genes.
Nat
Prod
Rep
2008;
25:188-‐200
[2] Shcherbakova
A,
Koptina
A,
Ulrich-‐Merzenich
G.
Rev
Clin
Pharm
Drug
Ther
Suppl.
2015:
84
[3] Kumar
J,
Dhar
P,
Tayade
AB,
Gupta
D,
Chaurasia
OP,
Upreti
DK
et
al.
Antioxidant
capacities,
phenolic
profile
and
cytotoxic
effects
of
saxicolous
lichens
from
trans-‐Himalayan
cold
desert
of
Ladack.
PLOS
ONE
2014;
9:
e98696
[4] Ghate
NB,
Chaudhuri
D,
Sarkar
R,
Sajem
AL,
Panja
S,
Rout
J,
Mandal
N.
An
antioxidant
extract
of
tropical
lichen,
Parmotrema
reticulatum,
induces
cell
cycle
arrest
and
apoptosis
in
breast
carcinoma
cell
line
MCF-‐7.
PLOS
ONE
2013;
8:
e82293
[5] Chou
T-‐C.
Drug
combination
studies
and
their
synergy
quantification
using
the
Chou-‐
Talalay
method.
Cancer
Res
2010;
70:
440-‐446
[6] Rosén
J.
Rickardson
L,
Backlund
A,
Gulbo
J,
Bohlin
L,
Larsson
R,
Gottfries
J,
ChemGPS-‐NP
mapping
of
chemical
compounds
for
prediciton
of
anticancer
mode
of
action.
QSAR
and
Comb
Chem
2009;
28:
436-‐440
P466
Isolation
and
characterization
of
lupane-‐type
triterpenes
from
Careya
arborea
and
their
effect
on
modulation
of
pro-‐
inflammatory
mediators
Rayhana
Begum1,
Manju
Sharma1,
Showkat
R.
Mir2
1
Department
of
Pharmacology,
2
Department
of
Pharmacognosy
and
Phytochemistry,
Faculty
of
Phar-‐
macy, Hamdard University, New Delhi-‐110 062, India
Careya arborea Roxb., belonging to family Lecythidaceae, is a large tree found throughout
India in deciduous forests and grassland [1] C. arborea is traditionally used for tumors, in-‐
flammation, bronchitis, epileptic fits, skin diseases, diarrhea, dysentery with bloody stools,
dyspepsia, ulcers, tooth ache and ear pain [2]. The stem bark is rich in triterpenes, including
betulinic acid [3]. Betulinic acid is a lupane triterpenoid, which has been reported to be a po-‐
tent blocker of many pro-‐inflammatory mediators such as NO (nitric oxide), MPO (myelop-‐
eroxidase), PGE2 (prostaglandin E2), TNF (tumor necrosis factor) and IL (interleukin) and
others [4]. Bio-‐activity guided chromatographic fractionation of the methanolic extract of C.
arborea bark resulted in isolation of six lupane-‐type triterpenes, characterized as: 3β-‐
hydroxy-‐lup-‐20 (29),21-‐trien-‐28-‐oic acid (1); 1, 3, 13, 16-‐tetrahydroxy-‐lup-‐9(11), 20(29)-‐
diene-‐28-‐oic acid (2); 1, 7-‐dihydroxy betulinic acid (3); 3β-‐O-‐dihydrocinnamyl betulinic acid
(4); 3β-‐O-‐trans-‐coumaryl lup-‐6, 9(11), 20(29)-‐triene-‐27, 28-‐dioic acid (5); and 16β-‐hydroxy-‐
2, 3-‐seco-‐lup-‐5, 20(29)-‐dien-‐2, 3, 28-‐trioic acid (6). The isolated compounds were character-‐
ized on the basis of FTIR, 1H NMR, 13C NMR, DEPT, COSY and MS techniques. This is the first
report of these compounds from the plant. Compounds 4 and 5, the novel phenyl propanoid
derivatives of betulinic acid, were found to be the most active when investigated with respect
to modulation of pro-‐inflammatory mediators such as NO, MPO, COX-‐2, TNF-‐α, IL-‐1β. The re-‐
sults indicated that the presence of phenyl propanoid functionality has profound effect on
down regulation of pro-‐inflammatory factors, even better than betulinic acid. The fact that
compounds with hydroxyl group at the para position of the phenyl ring are selective COX-‐2
inhibitors [5], further explains the better anti-‐inflammatory activity of these compounds.
Acknowledgements:
Jamia
Hamdard,
Delhi
is
acknowledged
for
providing
various
research
and
infra-‐
structural
facilities.
Keywords: Careya arborea; betulinic acid; lupane triterpenes, inflammation
References:
[1] Parinitha
M,
Harish
GU,
Vivek
NC,
Mahesh
T,
Shivanna
MB.
Ethnobotanical
wealth
of
Bhadra
wild
life
sanctuary
in
Karnataka.
Indian
J
Trad
Knowledge
2004;
3:
37-‐50
[2] Rayhana
B,
Sharma
M,
Pillai
KK,
Aeri
V,
Sheliya
MA.
Inhibitory
effect
of
Careya
arborea
on
inflammatory
biomarkers
in
carrageenan-‐induced
inflammation.
Pharm
Biol
2015;
53:
437-‐445
[3] Mahato
SB,
Kundu
AP.
13C
NMR
Spectra
of
pentacyclic
triterpenoids–a
compilation
and
some
salient
features.
Phytochemistry
1994;
37:
1517-‐1573
[4] Tsai
JC,
Peng
WH,
Chiu
TH,
Lai
SC,
Lee
CY.
Anti-‐inflammatory
effects
of
Scoparia
dulcis
L.
and
betulinic
acid.
Am
J
Chin
Med
2011;
39:
943-‐956
[5] Recio
MC,
Giner
RM,
Manez
S,
Gueho
J,
Julien
HR,
Hostettmann
K,
Ríos
JL.
Investigations
on
the
steroidal
anti-‐inflammatory
activity
of
triterpenoids
from
Diospyros
leucomelas.
Plan-‐
ta
Med
1995;
61:
9-‐12
P467
Cytotoxic
ambuic
acid
derivatives
from
Hawaiian
endophytic
fungus
FT-‐172
Chun-‐Shun
Li1,2,
Bao-‐Jun
Yang2,
James
Turkson2,
Shugeng
Cao1,2
1
Natural
Products
and
Experimental
Therapeutics,
Cancer
Centre,
University
of
Hawaii,
701
Ilalo
Street,
Honolulu,
Hawaii
96813,
USA;
2
Department
of
Pharmaceutical
Sciences,
Daniel
K.
Inouye
College
of
Pharmacy,
University
of
Hawai'i
at
Hilo,
200
West
Kawili
Street,
Hilo,
HI
96720,
USA
Five
new
ambuic
acid
derivatives,
pestallic
acids
A‒E
(1‒5)
and
three
known
analogs
(6‒8)1-‐3
were
isolated
from
the
cultured
broth
of
Pestalotiopsis
sp.
FT172.
The
structures
of
the
new
compounds
were
determined
through
the
analysis
of
HRMS
and
NMR
spectroscopic
data.
The
absolute
configurations
of
the
new
compounds
were
assigned
by
comparison
of
the
experi-‐
mental
and
calculated
electric
circular
dichroism
(ECD)
spectrum,
and
also
by
comparison
of
the
ECD
with
those
in
the
literature.
All
the
compounds
were
tested
against
A2780S
and
cis-‐
platin
resistant
A2780CisR
cell
lines,
and
compounds
5
and
7
showed
potent
activities
with
IC50
values
from
3.3
to
17.0
µM.
O O
18 OH
17 15 13 11
9
10
1 OH OH
5 3 O
O 19 O O
7
OH
R1 R2
O O
1 2 R1 =OH, R 2=H
6 R1 +R 2=O
OH R1 R2
OH OH
OH
O
O HO O
R1
R3 R4
O
5 R1 +R 2=O, R+R 4=O
3 R1 =OH 7 R1 +R 2=O, R 3=OH, R 4=H
4 R1 =H 8 R1 =R3=OH, R 2=R 4=H
Acknowledgements:
This
work
was
financially
supported
mainly
by
a
start-‐up
funding
from
University
of
Hawaii
Cancer
Centre
and
the
Victoria
S.
and
Bradley
L.
Geist
Foundation
(15ADVC-‐74420)
to
SC.
We
are
grateful
to
Mr.
Wesley
Yoshida
and
Dr.
Walter
Niemczura
at
the
Chemistry
Department,
University
of
Hawaii
at
Manoa
for
collecting
the
NMR
data.
Keywords: Anticancer, Hawaii, Endophytic fungi, MS, CD, NMR
References:
[1] Xie
J,
Li
J,
Yang
YH,
Chen
YH,
Zhao
PJ.
Two
new
ambuic
acid
analogs
from
Pestalotiopsis
sp.
cr013.
Phytochem
Lett
2014;
10:
291-‐294
[2] Li
JY,
Harper
JK,
Grant
DM,
Tombe
BO,
Bashyal
B,
Hess
WM,
Strobel
GA.
Ambuic
acid,
a
high-‐
ly
functionalized
cyclohexenone
with
antifungal
activity
from
Pestalotiopsis
spp.
and
Monochaetia
sp.
Phytochemistry
2001;
56:
463-‐468
[3] Qi
QY,
Li
EW,
Han
JJ,
Pei
YF,
Ma
K,
Bao
L,
Huang
Y,
Zhao
F,
Liu
HW.
New
ambuic
acid
deriv-‐
atives
from
the
solid
culture
of
Pestalotiopsis
neglecta
and
their
nitric
oxide
inhibitory
ac-‐
tivity
Sci
Rep
2015;
5:
9958
P468
Alpine
plants
with
anti-‐ageing
effects
Silvia
Revoltella1,
Birgit
Waltenberger1,
Giorgia
Baraldo2,
Clemens
Kohl3,
Pagitz
Konrad4,
Ro-‐
land
Kohl3,
Pidder
Jansen-‐Dürr2,
Hermann
Stuppner1
1Institute
of
Pharmacy/Pharmacognosy
and
Center
for
Molecular
Biosciences
Innsbruck
(CMBI),
Univer-‐
sity
of
Innsbruck,
Innrain
80-‐82,
6020
Innsbruck,
Austria,
2Research
Institute
for
Biomedical
Aging
Re-‐
search
and
CMBI,
University
of
Innsbruck,
Rennweg
10,
6020
Innsbruck,
Austria,
3Cura
Marketing
GmbH,
Dr.-‐Franz-‐Werner-‐Straße
19,
6020
Innsbruck,
Austria,
4Institute
of
Botany,
University
of
Innsbruck,
Sternwartestraße
15,
6020
Innsbruck,
Austria
Alpine
plants
represent
a
valuable
source
for
the
identification
of
novel
bioactive
natural
products
with
anti-‐ageing
effects.
Due
to
the
harsh
conditions
present
in
higher
altitudes,
they
accumulate
certain
secondary
metabolites
in
larger
quantities
[1],
especially
substances
that
protect
plants
against
UV
radiation
[2],
which
is
known
to
contribute
to
the
ageing
of
human
skin
[3,
4].
Therefore,
Alpine
plants
provide
enormous
potential
for
the
development
of
new
cosmeceuticals,
i.e.,
cosmetics
with
proven
pharmacological
effects.
Within
this
project,
150
plant
species,
mainly
from
the
Alpine
region,
are
selected,
collected,
and
extracted
successively
with
EtOAc
and
MeOH.
Extracts
are
investigated
for
their
ability
(i)
to
inhibit
NADPH
oxidase
4
(NOX4)
in
HEK
293
cells,
(ii)
to
activate
proteasome
in
HEK
293
cells,
and
(iii)
to
inhibit
tyrosinase
in
a
TLC
autographic
assay.
Moreover,
their
phytochemical
profile
is
being
analysed
using
TLC
and
HPLC-‐MS.
So
far,
out
of
the
50%
of
the
plants,
which
were
already
tested
on
NOX4
inhibition,
Alnus
viridis
(leaf
MeOH
extract),
Aruncus
dioicus
(lower
parts
MeOH
and
EtOAc
extracts),
and
Berberis
ilicifolia
(stem
and
leaf
MeOH
extracts)
were
identified
as
promising
plants
with
significant
inhibitory
activity
on
NOX4
at
a
concen-‐
tration
of
25
µg/mL,
showing
80,
94,
and
94%
luminescence
quenching,
respectively.
Cell
via-‐
bility
measurements
revealed
that
these
extracts
are
not
cytotoxic
in
the
tested
concentration.
While
the
tests
on
proteasome
activation
are
still
ongoing,
all
plant
extracts
were
already
tested
on
tyrosinase
inhibition
in
a
bioautographic
assay.
Therein,
14%
showed
a
good
inhibi-‐
tory
activity.
Further
studies
will
address
the
identification
of
the
bioactive
constituents
and
the
determination
of
their
molecular
mode
of
action.
The
final
goal
of
this
project
is
the
identi-‐
fication
and
generation
of
plant
extracts
with
anti-‐ageing
activity
in
vitro
and
in
vivo,
which
can
be
used
for
the
development
of
cosmeceuticals.
Acknowledgements:
Supported
by
the
Austrian
Forschungsförderungsgesellschaft
(FFG),
(BRIDGE
1
pro-‐
ject
848474)
Keywords: Alpine plants, cosmeceuticals, anti-‐ageing, NOX4, proteasome, tyrosinase
References:
[1] Zidorn
C.
Altitudinal
variation
of
secondary
metabolites
in
flowering
heads
of
the
Aster-‐
aceae:
trends
and
causes.
Phytochem
Rev
2010;
9:
197−203
[2] Debacq-‐Chainiaux
F,
Leduc
C,
Verbeke
A,
Toussaint
O.
UV,
stress
and
aging.
Dermatoendo-‐
crinol
2012;
4:
236−240
[3] Yaar
M,
Gilchrest
BA.
Photoageing:
mechanism,
prevention
and
therapy.
Br
J
Dermatol
2007;
157:
874−887
[4] Turunen
M,
Latola
K.
UV-‐B
radiation
and
acclimation
in
timberline
plants.
Environ
Pollut
2005;
137:
390−403
P469
Triterpensaponins
as
a
novel
transfection
enhancer
Simko
Sama,
Matthias
F.
Melzig,
Alexander
Weng
Department
of
Pharmaceutical
Biology,
Institute
of
Pharmacy,
Freie
Universität
Berlin,
Königin-‐Luise-‐
Straße
2-‐4,
14195
Berlin,
Germany
Recent
studies
have
shown
that
triterpensaponins
of
Saponaria
officinalis
L.
(Caryophyllace-‐
ae)
and
Gypsophila
paniculata
L.
(Caryophyllaceae)
are
able
to
augment
the
effect
of
certain
toxins
such
as
ribosome
inactivating
proteins
(RIPs)
[1].
Due
to
their
amphiphilic
properties
these
saponins
are
able
to
interact
with
the
endosome
and
lysosome
membrane
and
thus
en-‐
hance
the
release
of
the
cargo
into
the
cytosol
[2].
In
order
to
exploit
this
kind
of
interaction,
in
this
study
DNA
containing
nanoparticles
were
formulated
and
characterized
to
be
applied
together
with
saponins.
Murine
neuroblastoma
cells
were
transfected
with
nanoplexes,
con-‐
taining
mammalian
plasmids
with
green
fluorescing
protein
(GFP),
Luciferase
or
the
RIP
Sap-‐
orin
of
Saponaria
officinalis
L.
A
transfection
with
saponins
showed
a
significant
increase
of
transfection
efficiency
in
terms
of
fluorescence,
luminescence
or
viability
compared
to
a
transfection
without
saponin
co-‐application.
First
in-‐vitro
cell
experiments
showed
a
6-‐fold
increase
of
luciferase
expression
compared
to
nanoparticles
alone.
GFP-‐transfection
even
achieved
a
33-‐fold
increase
of
GFP-‐expression.
With
the
co-‐application
of
saponins
a
transfec-‐
tion
of
52%
regarding
all
cells
could
be
observed,
while
nanoparticles
just
showed
an
efficien-‐
cy
of
2%.
Therefore
triterpensaponins
hold
a
great
potential
to
improve
the
transfection
effi-‐
ciency
and
for
being
used
as
a
valuable
transfection
booster
in
pharmaceutical
therapies.
Keywords: Transfection, Triterpensaponins
References:
[1] Weng
A,
Thakur
M,
Beceren-‐Braun
F,
Bachran
D,
Bachran
C,
Riese
SB,
Jenett-‐Siems
K,
Gilabert-‐Oriol
R,
Melzig
MF,
Fuchs
H.
The
toxin
component
of
targeted
anti-‐tumor
toxins
determines
their
efficacy
increase
by
saponins.
Mol
Oncol
2012;
6:
323-‐332
[2] Weng
A,
Thakur
M,
von
Mallinckrodt
B,
Beceren-‐Braun
F,
Gilabert-‐Oriol
R,
Wiesner
B,
Eichhorst
J,
Böttger
S,
Melzig
MF,
Fuchs
H.
Saponins
modulate
the
intracellular
trafficking
of
protein
toxins.
J
Control
Release
2012;
164:
74-‐86
P470
Autoxidation
of
Betulonaldehyde
Sloan Ayers, Tamas Benkovics, Jonathan Marshall, Yichen Tan
Pharmaceutical Development, Bristol-‐Myers Squibb Co., 1 Squibb Dr., New Brunswick, NJ 08903, USA
Betulonaldehyde
is
a
pentacyclic
triterpenoid
found
in
Betula
sp.
as
a
minor
constituent
along
with
the
major
secondary
metabolites
betulin
and
betulinic
acid.
In
a
stability
study,
betu-‐
lonaldehyde
was
stirred
in
acetone
at
room
temperature
under
air.
Three
major
degradants
were
detected,
and
were
isolated
after
12
days.
HRMS
showed
that
for
one
of
the
degradants,
a
single
oxygen
atom
had
been
added,
and
this
product
was
confirmed
by
HPLC
(retention
time
match
to
an
authentic
standard)
and
NMR
to
be
betulonic
acid.
The
remaining
two
degradants
also
gained
oxygen,
but
lost
a
carbon.
HRMS
and
NMR
data
showed
that
these
degradants
were
C17
hydroperoxide
epimers.
The
identity
of
the
stereochemical
configura-‐
tion
at
C17
for
each
these
hydroperoxides
was
deduced
by
2D-‐NOESY
correlations
and
the
coupling
pattern
for
H18.
Acknowledgements: Drs. Charles Pathirana and Scott Miller are acknowledged for helpful discussions.
Keywords: betulonaldehyde, autoxidation, hydroperoxide
P471
Laricifomes
officinalis
–
a
rich
source
of
pharmacologically
active
triterpenes
Sonja
Sturm,
Katharina
Gallmetzer,
Andreas
Friedl,
Birgit
Waltenberger,
Veronica
Temml,
Hermann
Stuppner
Institute
of
Pharmacy/Pharmacognosy,
Center
for
Molecular
Biosciences,
University
of
Innsbruck,
Innrain
80-‐82,
Innsbruck,
Austria
Laricifomes
officinalis,
(Vill.)
Kotl.
&
Pouzar
(Polyporaceae)
is
a
wood
rotting
fungus
growing
on
living
or
dead
conifers,
particularly
on
larches.
It
is
also
known
as
agarikon,
quinine
conk,
or
in
German
speaking
regions
as
“Apothekerschwamm”
referring
to
its
relevance
in
tradi-‐
tional
medicine.
L.
officinalis
might
be
considered
as
the
most
common
and
versatile
used
me-‐
dicinal
fungus
of
the
past,
used
to
treat
tuberculosis,
asthma
and
cough,
night
sweats,
as
laxa-‐
tive,
or
as
bitter
to
aid
digestion
[1,2].
In
the
course
of
an
ongoing
project
aiming
at
the
search
for
new
therapeutic
treatment
options
for
metabolic
syndrome,
extracts
of
L.
officinalis
were
tested
in
a
protein
tyrosine
phosphatase
1B
(PTB1B)
assay.
This
enzyme
is
a
key
regulator
of
insulin-‐receptor
activity
reversing
the
autophosphorylation
of
the
receptor,
and
also
acting
at
downstream
signalling
components.
As
an
elevated
PTP1B
expression
can
be
associated
with
obesity
and
insulin
resistance,
PTP1B
is
discussed
as
possible
new
target
for
the
treatment
of
diabetes
type
2
and
obesity
[3].
A
methanolic
and
an
ethyl
acetate
extract
showed
significant
inhibitory
activity
on
PTP1B
(>85%
at
30
µg/mL).
A
first
fast
fractionation
of
the
ethyl
acetate
extract
revealed
that
agaricinic
acid
(82%
at
30
µg/mL),
as
well
as
a
terpenoid
containing
fraction
(84%
at
30
µg/mL)
distinctively
contributed
to
this
activity.
Furthermore,
known
constituents
of
L.
officinalis
[2]
were
screened
with
two
sets
of
structure-‐based
pharmaco-‐
phore
models
(Discovery
Studio
and
Ligandscout)
resulting
in
the
identification
of
several
triterpene
derivatives
as
potential
PTP1B
inhibitors.
These
findings
led
to
the
phytochemical
investigation
of
the
terpene-‐profile
of
this
fungus.
In
a
first
step
a
HPLC-‐DAD-‐APCI-‐MS/MS
method
was
developed
for
the
comprehensive
profiling
and
characterisation
of
the
terpenoids
in
L.
officinalis.
The
following
phytochemical
work-‐up
of
the
extract
resulted
in
the
isolation
and
structural
characterisation
of
the
sesquiterpene
lacrinolic
acid
and
24
lanostane
triter-‐
penes.
Nine
compounds
were
already
known
as
constituents
from
this
fungus,
seven
triter-‐
penes
are
described
the
first
time
for
L.
officinalis,
and
further
eight
triterpenes
were
identi-‐
fied
as
new
natural
products.
Evaluation
of
the
isolated
compounds
for
their
inhibitory
effects
on
PTP1B
is
in
progress.
Keywords:
Laricifomes
officinalis,
Polyporaceae,
Protein
tyrosine
phosphatase
1B,
Lanostane-‐
type
triterpenes,
HPLC-‐DAD-‐APCI-‐MS/MS
References:
[1] Molitoris
HP.
Pilze
in
Medizin,
Folklore
und
Religion.
Feddes
Repertorium
2002;
113:
165-‐
182
[2] Grienke
U,
Zöll
M,
Peintner
U,
Rollinger
JM.
European
medicinal
polypores
–
A
modern
view
on
traditional
uses.
J
Ethnopharmacol
2014;
154:
564-‐583
[3] Johnson
TO,
Ermolieff
J,
Jirousek
MR.
Protein
Tyrosine
Phosphatase
1b
Inhibitors
for
diabe-‐
tes.
Nat
Rev
Drug
Discov
2002;
1:
696-‐709
P472
Anti-‐amyloidogenic
constituents
from
roots
of
Dryopteris
cras-‐
sirhizoma
in
Alzheimer’s
disease
cellular
model
Hwan-‐Bin
Joo1,
Jae-‐Moon
Kim2,
So-‐Young
Park1,2
1
Laboratory
of
Pharmacgnosy,
College
of
Pharmacy,
Dankook
University,
Cheonan
330-‐714,
Republic
of
Korea,
2
World
Class
University,
Department
of
Nanobiomedicine,
Dankook
University,
Cheonan
330-‐714,
Korea
β-‐amyloid
(Aβ),
one
of
the
main
causes
of
Alzheimer’s
disease
(AD)
[1],
is
generated
by
the
amyloidogenic
pathway
from
amyloid
precursor
protein
(APP)
by
β-‐secretases
and
γ-‐
secretases
[2].
The
generated
Aβ
monomers
aggregate
into
oligomers
and
fibrils,
and
induce
neurotoxicity
and
neuronal
cell
death
[3].
Thus,
we
investigated
the
anti-‐amyloidogenic
com-‐
pounds
from
natural
sources
using
Chinese
Hampster
Ovary
(CHO)
cells
stably
over-‐
expressing
APP
(APP-‐CHO
cells).
The
methanol
extract
of
Dryopteris
crassirhizoma
Nakai
[Dryopteridaceae]
roots
efficiently
reduced
the
Aβ
production.
The
reduction
was
more
pro-‐
nounced
by
the
n-‐butanol
layer
of
the
extract.
Active
constituents
inhibiting
Aβ
production
were
isolated
by
activity-‐guided
fractionation.
Nine
compounds
were
isolated
including
the
flavonoids
epicatechin
(1),
β-‐carboxylmethyl-‐(-‐)-‐epicatechin
(2),
eriodictyol
(5),
the
chro-‐
mones
7-‐methoxy-‐isobiflorin
(3),
biflorin
(4)
and
noreugenin
(6),
and
the
phloroglucinols
butyrylphloroglucinol
(7),
2-‐propionyl-‐4-‐methylphloroglucinol
(8)
and
2-‐butyryl-‐4-‐methyl-‐
phloroglucinol
(9).
All
compounds
were
tested
for
the
effects
on
the
production
of
Aβ
by
ELI-‐
SA
and
sAPPβ
by
Western
blot
analysis.
Compounds
3-‐9
significantly
inhibited
the
production
of
Aβ.
Particularly,
20
μg/mL
of
compound
7
and
9
reduced
80%
of
Aβ
production
compared
to
controls
(IC50
=
9.8
and
5.2
μg/mL,
respectively).
In
addition,
compounds
3-‐9
also
signifi-‐
cantly
reduced
the
production
of
sAPPβ.
Compound
7
and
9
(20
μg/mL)
reduced
the
sAPPβ
production
by
60
and
80%
compared
to
the
controls,
respectively.
Furthermore,
these
results
were
accompanied
with
the
reduced
protein
expression
of
β-‐secretases.
Compound
7
and
9
(20
μg/mL)
reduced
about
30%
of
the
expression
of
β-‐secretases
compared
to
the
controls.
The
results
suggest
that
compounds
3-‐9
significantly
reduced
the
Aβ
production
by
inhibiting
β-‐secretases
and
if
they
can
cross
the
blood-‐brain
barrier
might
be
beneficial
for
AD
as
pre-‐
ventatives
or
therapeutics.
Acknowledgements:
This
work
was
supported
by
the
research
fund
of
the
National
Research
Foundation
of
Korea
(NRF-‐2010-‐000398,
NRF-‐2011-‐000398).
Keywords: Dryopteris crassirhizoma, beta-‐amyloid, Alzheimer’s disease, beta-‐secretase
References:
[1] Tanzi
RE,
Moir
RD,
Wagner
SL.
Clearance
of
Alzheimer's
Abeta
peptide:
the
many
roads
to
perdition.
Neuron
2004;
43:
605-‐608
[2] Zhang
H,
Ma
Q,
Zhang
YW,
Xu
H.
Proteolytic
processing
of
Alzheimer's
beta-‐amyloid
pre-‐
cursor
protein.
J
Neurochem
2012;
120
(Suppl
1):
9-‐21
[3] Kawahara
M,
Kuroda
Y.
Molecular
mechanism
of
neurodegeneration
induced
by
Alzhei-‐
mer's
beta-‐amyloid
protein:
channel
formation
and
disruption
of
calcium
homeostasis.
Brain
Res
Bull
2000;
53:
3893-‐3897
P473
Anti-‐inflammatory
activity
of
a
Carica
papaya
leaf
extract
used
to
prevent
occasional
gingivitis
Stéphane
Dejoie1,2,
Rim
Boulmane1,
Séverine
Derbré1,
Daniel
Henrion3,
Christophe
Binachon2,
Pascal
Richomme1
1
EA921
SONAS/SFR4207
QUASAV,
Université
d’Angers,
France;
2
SAS
ESPRIT
D’ETHIQUE,
11
av.
Félix
Vincent,
Orvault,
France;
3
BNMI,
UMR
Inserm
1083,
UMR
CNRS
6214,
Angers,
France
Gencix®
made
from
a
Carica
papaya
(pawpaw)
aqueous
leaf
extract
mixed
with
micronized
pumice
stone,
is
a
toothpaste
proposed
to
protect
gums
and
prevent
occasional
gingivitis.
In
order
to
understand
the
role
of
the
natural
products
(NPs)
from
C.
papaya
leaves
responsible
for
the
positive
effects
observed
by
dentists
using
this
product
during
their
clinical
practice,
we
focused
in
this
work
on
their
anti-‐inflammatory
potential
[1,2].
After
liquid-‐liquid
extraction
of
the
aqueous
leaf
extract
with
EtOAc,
the
effect
on
inflamma-‐
tion
of
the
resulting
aqueous
and
organic
extracts
was
evaluated
by
monitoring
the
release
of
tumor
necrosis
factor-‐alpha
(TNF-‐α),
a
pro-‐inflammatory
cytokine,
by
human
monocytic
cells
derived
from
an
acute
monocytic
leukemia
(THP-‐1)
following
a
Porphyromonas
gingivalis
LPS
activation.
The
EtOAc
extract
(inhibition
at
500
µg/mL:
60.2%)
was
active
when
compared
with
dexamethasone
used
as
the
reference
compound,
whereas
a
lower
activity
was
detected
for
the
aqueous
extract
(inhibition
at
500
µg/mL:
38.3%)
(Figure
1).
A
bioguided
fractiona-‐
tion
of
the
EtOAc
extract
was
thus
undertaken
to
isolate
the
anti-‐inflammatory
PNs.
A
derepli-‐
cation
study
was
also
conducted
using
classical
HPLC-‐DAD-‐MSn
as
well
as
13C
NMR
dereplica-‐
tion
as
previously
described
by
J.
Hubert
et
al.
[3].
Caffeoyl
malate
and
their
derivatives
were
identified
as
major
NPs
of
the
EtOAc
extract
but
were
not
found
responsible
for
the
anti-‐
inflammatory
potential.
Flavonols
such
as
quercetin
and
kaemperol
and
their
glycosides
were
identified
as
minor
compounds
of
the
EtOAc
extract.
Anti-‐TNF-‐α
activity
of
C.
papaya
leaf
ex-‐
tract
is
probably
related
to
flavonols
as
already
observed
in
other
species
such
as
Ximenia
caffra
[4]
and
thus
maybe
responsible
for
the
anti-‐inflammatory
effect
of
active
fractions.
This
activity
could
partly
explain
the
positive
effects
of
Gencix®
observed
by
dentists.
Keywords: Carica papaya, Anti TNF-‐α, Toothpaste
References:
[1] Njoku
OV,
Obi
C.
Phytochemical
constituents
of
some
selected
medicinal
plants.
African
J
Pure
Appl
Chem
2009;
3:
228-‐233
[2] Owoyele
BB
V,
Adebukola
OMO,
Funmilayo
AA,
Soladoye
AO.
Anti-‐inflammatory
activities
of
ethanolic
extract
of
Carica
papaya
leaves.
Inflammopharmacology
2008;
16:
168-‐173
[3] Hubert
J,
Chollet
S,
Purson
S,
Reynaud
R,
Harakat
D,
Martinez
A,
et
al.
Exploiting
the
Com-‐
plementarity
between
Dereplication
and
computer-‐assisted
structure
elucidation
for
the
chemical
profiling
of
natural
cosmetic
ingredients:
Tephrosia
purpurea
as
a
case
study.
J
Nat
Prod
2015;
78:
1609-‐1617
[4] Zhen
J,
Guo
Y,
Villani
T,
Carr
S,
Brendler
T,
Mumbengegwi
DR,
Kong
ANT,
Simon
JE,
Wu
Q.
Phytochemical
analysis
and
anti-‐inflammatory
activity
of
the
extracts
of
the
African
me-‐
dicinal
plant
Ximenia
caffra.
J
Anal
Methods
Chem
2015;
2015:
948262
70,0
60,2
TNF-‐alpha
Inhibi.on
60,0
55,1
50,0
38,3
40,0
30,0
27,1
26,6
16,2
16,9
20,0
10,0
1,5
0,0
3,1
4,4
0,0
-‐10,0
GCX
H20
EtOAc
DMSO
DEXA
1µg/ml
Figure
1:
Inhibition
(%)
of
TNF-‐α
release
by
Gencix®
(GCX)
and
its
aqueous
(H2O)
and
ethyl
acetate
(EtOAc)
extracts
obtained
after
liquid
liquid
partition.
500µg/mL
(red);
150
µg/mL
(light
blue);
50µg/mL
(green).
References
(dark
blue):
Dexamethasone
(DEXA,
1µg/mL);
DMSO
(0.1%)
P474
Potential
immunomodulatory
compounds
isolated
from
red
betel
leaves
(Piper
crocatum,
Ruiz
&
Pav.)
Subagus
Wahyuono1,
Paula
Kustiawan2,
Yustina
Sri
Hartini3,
Yuswanto1,
Sitarina
Widyarini4
1
Faculty
of
Pharmacy,
Universitas
Gadjah
Mada
-‐
Yogyakarta,
2
Faculty
of
Forestry,
Universitas
Mula-‐
warman
–
Samarinda,
3
Faculty
of
Pharmacy,
Universitas
Sanata
Dharma
–
Yogyakarta,
4
Faculty
of
Veterinery
Medicine,
Universitas
Gadjah
Mada
–
Yogyakarta,
INDONESIA
Red
betel
leaves
(Piper
crocatum,
Ruiz.
&
Pav.)
have
been
traditionaly
used
as
an
ingredient
in
a
Jamu
preparation
intended
to
maintain
health.
The
red
betel
leaves
are
thought
to
boost
the
immune
system,
keeping
the
body
healthy.
The
study
aimed
to
isolate
and
identify
com-‐
pounds
that
stimulate
the
immune
system,
and
determine
their
potential
as
immunomodula-‐
tors
in
vivo.
Isolation
of
the
active
compounds
was
done
through
bioassay-‐guided
isolation.
Activation
of
macrophage
in
vitro
(phagocytic
assay)
was
used
to
guide
the
isolation
of
active
compounds.
The
isolated
active
compounds
were
tested
for
in
vivo
immunomodulatory
effect
on
balb/c
mice
infected
with
Lysteria
monocytogenes.
In
these
assays
were
measured,
phago-‐
cytic
index
and
capacity,
concentration
of
nitric
oxide
and
proliferation
of
lymphocyte.
Two
new
neolignans,
2-‐allyl-‐4-‐(1’-‐hydroxy-‐1’-‐(3”,4”,5”-‐trimethoxyphenyl)propan-‐2’-‐yl)-‐3,5-‐
dimethoxycyclohexa-‐2,5-‐dienone
(1)
and
its
naturally
occuring
acetyl
derivative
(2)
were
isolated
and
identified
as
active
compounds.
Compounds
1
and
2
separately
(5
µg/mL)
displayed
immunomodulatory
activity
equal
to
Imboost
(Echinaceae
product),
as
positive
control.
In
the
in
vivo
immunomodulatory
assay,
1
and
2
separately
(5.0
and
10.0
mg/kgBW)
increased
the
activity
and
capacity
of
macrophages.
In
addition,
increases
of
nitric
oxide
and
IL-‐12
production
were
also
observed
in
response
to
to
1
and/or
2
(2.5,
5.0
and
10.0
mg/kgBW).
No
lymphocyte
proliferation
was
observed,
which
might
be
due
to
inhibition
on
the
binding
between
IL-‐12
and
its
receptor.
Keywords:
Piper
crocatum,
Ruiz.
&
Pav.,
immunomodulatory,
macrophages,
nitric
oxide,
IL-‐
12
1
2
P475
The
potential
of
Combretum
molle
in
the
treatment
of
skin
cancer
Sunelle
Rademan,
Namrita
Lall
Department of Plant Sciences, University of Pretoria, Hatfield campus, Pretoria, South Africa
Cancer
is
a
group
of
diseases
occurring
in
all
regions
of
the
world
and
is
the
cause
of
millions
of
deaths
each
year.
It
is
believed
that
approximately
132
000
melanoma
skin
cancer
cases
arise
each
year
[1].
This
high
incidence
of
skin
cancer
cases
in
mostly
associated
with
in-‐
creased
levels
of
ultraviolet
radiation
[2].
Plants
provide
an
immeasurable
source
of
bioactive
compounds
for
drug
discovery.
Some
of
these
plant-‐derived
compounds
are
currently
being
used
in
chemotherapy
for
the
treatment
of
a
variety
of
cancers
[3].
The
vast
amount
of
unex-‐
plored
plants
therefore
represents
the
potential
of
finding
new
drugs
for
the
treatment
of
cancer.
The
current
study
focuses
on
the
ethanolic
leaf
and
fruit
extracts
of
Combretum
molle
R.
Br.
ex
G.
Don
[Combretaceae]
and
the
potential
of
these
extracts
for
anti-‐cancer
activity
against
skin
cancer
and
their
possible
mechanisms
of
action.
The
C.
molle
extracts
were
inves-‐
tigated
for
their
cytotoxicity
and
synergistic
activity
on
two
human
skin
cancer
cell
lines
(MEL-‐1;
A431)
and
a
non-‐cancerous
skin
cell
line
(HaCat)
using
the
XTT
Cell
Proliferation
Kit
II.
Cell
death
studies
of
the
lead
extracts
were
determined
using
light
microscopy
and
an
An-‐
nexin
V
kit.
Other
mechanistic
studies
included
inflammation
and
immunity
studies
and
the
anti-‐metastatic
potential
of
the
lead
extracts.
The
leaf
extract
of
C.
molle
showed
the
highest
cytotoxic
activity
on
the
MEL-‐1
cells
with
an
IC50
of
17.56
µg/ml.
The
leaf
and
fruit
extracts
of
Combretum
molle
showed
good
activity
on
the
A431
cell
line
with
IC50
values
of
27.03
µg/ml
and
29.99
µg/ml,
respectively.
From
the
light
microscopy
images
morphological
characteris-‐
tics
of
apoptosis
were
observed.
Both
the
fruit
and
leaf
extracts
also
showed
significant
anti-‐
inflammatory
potential.
The
results
obtained
for
the
C.
molle
extracts
indicate
that
these
ex-‐
tracts
can
be
potentially
useful
as
a
topical
application
for
the
treatment
of
squamous
cell
car-‐
cinoma
and
melanoma
skin
cancer.
Acknowledgements:
NRF
for
the
financial
assistance
Keywords: Cancer, skin cancer, plants, Combretum molle
References:
[1] WHO
Skin
cancer
fact
sheet.
www.who.int/uv/faq/skincancer/en/index1.html
accessed:
03
March
2014
[2] Cancer.org
Skin
cancer.
http://www.cancer.org/cancer/skincancer-‐
melanoma/detailedguide/melanoma-‐skin-‐cancer-‐treating-‐chemotherapy
accessed:
7
April
2015
[3] Nobili
S,
Lippi
D,
Witort
E,
Donnini
M,
Bausi
L,
Mini
E,
Capaccioli
S.
2009.
Natural
com-‐
pounds
for
cancer
treatment
and
prevention.
Pharmacological
research
2009,
59:
365-‐
378
P476
A
bioactivity-‐guided
screening
of
Sri
Lankan
plants
in
the
search
for
novel
antibacterial
and
anticancer
agents.
Supun
Mohotti1,2,
Sanjeevan
Rajendran1,
Taj
Muhammad1,
Adam
A.
Strömstedt1,
Robert
Bur-‐
man1,
Björn
Hellman3,
E.D.
de
Silva2,
Ulf
Göransson2,
C.M.
Hettiarachchi2,
Sunithi
Gunasekara1
1Division of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Biomedical Centre,
SE
75123
Uppsala,
Sweden,
2Department
of
Chemistry,
Faculty
of
Science,
University
of
Colombo,
Thurstan
Rd,
Colombo
00300,
Sri
Lanka
,3
Division
of
Toxicology
Department
of
Pharmaceutical
Biosci-‐
ences,
Uppsala
University,
Biomedical
Centre,
SE-‐741
24
Uppsala,
Sweden
Sri
Lankan
Ayurvedic
system
of
traditional
medicine
derives
from
plant-‐based
remedies
handed
down
from
generation
to
generation
over
a
period
of
3000
years.
Despite
being
an
integral
part
of
the
Sri
Lankan
culture,
the
scientific
evidence
for
the
biological
activity
of
the-‐
se
traditional
medicines
is
often
lacking
in
most
cases.
Concerns
are
related
to
lack
of
scien-‐
tific
evidence
for
efficacy,
toxicity,
standardization
of
formulations
as
well
as
adverse
interac-‐
tions
of
the
medications.
The
current
project
was
conducted
to
identify
and
characterize
chemical
compounds
present
in
selected
Sri
Lankan
medicinal
plants
that
have
potential
utili-‐
ty
as
therapeutic
candidates
for
antimicrobial
and
anti-‐cancer
applications.
We
have
screened
52
medicinal
plants
from
the
families
Fabaceae,
Cucurbitaceae,
Rubiaceae
and
Violaceae
for
antibacterial
activity
and
cytotoxicity
by
a
microfractionation
protocol.
Each
crude
plant
extract
was
fractionated
into
a
96
deep
well
microtiter
plate
using
semi-‐
preparative
HPLC
(2
mg
of
crude
extract
to
give
45
×
1
ml
fractions)
and
the
fractions
were
screened
against
Staphylococcus
aureus
in
an
optimized
Minimum
Inhibitor
Concentration
(MIC)
assay
[1].
A
Fluorometric
Microculture
Cytotoxic
Assay
(FMCA)
[2]
using
a
human
lym-‐
phoma
cell
line
was
conducted
to
identify
fractions
with
cytotoxic
activity.
The
corresponding
masses
in
the
bioactive
fractions
were
then
identified
by
LC-‐MS
analysis.
Currently,
we
have
isolated
seven
bioactive
molecules
in
the
mass
range
500-‐1000
Da
.
Acknowledgements:
International
Collaborative
Grant,
Swedish
Research
Council
Grant
Keywords: Ayurvedic, MIC, NMR, FMCA , antibacterial, anticancer,
References:
[1] Strömstedt
AA,
Kristiansen
PE,
Gunasekera
S,
Grob
N,
Skjeldal
L,
Göransson
U.
Selective
membrane
disruption
by
the
cyclotide
kalata
B7:
Complex
ions
and
essential
functional
groups
in
the
phosphatidylethanolamine
binding
pocket.
Biochim
Biophys
Acta.
2016;
1858:
1317-‐1327
[2] Lindhagen
E,
Nygren
P,
Larsson
R.
The
fluorometric
microculture
cytotoxicity
assay.
Nat
Protoc
2008;
3:
1364-‐1369.
P477
Identifying
specific
inhibitors
of
triple-‐negative
breast
cancer
subtypes
Andrew
J.
Robles1,
Shengxin
Cai3,4,
Lin
Du3,4,
Corena
V.
Shaffer1,
Tanja
Grkovic5,
April
L.
Risinger1,2,
Barry
R.
O’Keefe6,7,
Robert
H.
Cichewicz3,4,
Susan
L.
Mooberry1,2
1
Department
of
Pharmacology,
2
Cancer
Therapy
&
Research
Center,
The
University
of
Texas
Health
Sci-‐
ence
Center
at
San
Antonio,
7703
Floyd
Curl
Drive,
San
Antonio,
Texas,
78229,
USA.
3
Department
of
Chemistry
and
Biochemistry,
4
Natural
Products
Discovery
Group,
University
of
Oklahoma,
101
Stephen-‐
son
Parkway,
Norman,
Oklahoma,
73019,
USA.
5
Natural
Products
Support
Group,
Leidos
Biomedical
Re-‐
search
Inc.,
Frederick
National
Laboratory
for
Cancer
Research,
Frederick,
MD
21702.
6
Natural
Products
Branch,
Division
of
Cancer
Treatment
and
Diagnosis,
7
Molecular
Targets
Laboratory,
Center
for
Cancer
Research,
National
Cancer
Institute,
Frederick,
Maryland,
21702.
Triple
negative
breast
cancer
(TNBC)
is
a
highly
aggressive
form
of
the
disease
characterized
by
the
lack
of
expression
of
the
estrogen
(ER),
progesterone
(PR)
and
HER2
receptors,
which
occurs
predominately
in
younger
patients
and
those
of
African
American
descent.
These
pa-‐
tients
have
a
poor
prognosis
due
in
part
to
the
lack
of
therapies
to
specifically
target
these
tumor
types.
Previous
work
has
demonstrated
that
TNBC
is
a
heterogeneous
disease
made
up
of
5
distinct
sub-‐types
that
have
been
molecularly
classified
using
patient
samples:
basal
like
1
(BL1),
basal
like
2
(BL2),
mesenchymal-‐like
(M),
mesenchymal
stem-‐like
(MSL)
and
luminal
androgen
receptor
(LAR)
[1].
We
have
taken
a
novel
approach
of
screening
plant
and
fungal
extracts
against
cancer
cell
lines
representing
each
of
these
TNBC
subtypes
to
identify
sub-‐
type-‐specific
cytotoxic
compounds.
Extracts
selective
for
each
of
the
5
TNBC
subtypes
have
been
identified
and
bioassay-‐guided
fractionation
has
thus
far
yielded
5
classes
of
compounds
with
selective
activity
for
4
of
the
TNBC
subtypes.
Interestingly,
some
of
the
compounds
with
the
most
exquisite
selectivity
are
known
compounds;
however
their
selectivities
for
distinct
subtypes
of
breast
cancer
have
not
been
previously
reported.
While
some
of
the
isolated
com-‐
pounds
themselves
may
be
interesting
drug
leads
for
TNBC
subtypes,
others
have
informed
on
the
potential
use
of
approved
therapies
that
may
be
effective
against
specific
TNBC
sub-‐
types.
This
is
the
case
for
deguelin,
a
compound
with
selective
activity
against
the
LAR
TNBC
subtype,
which
has
dual
mTOR/androgen
receptor
inhibitory
activities,
suggesting
that
com-‐
binations
of
inhibitors
targeting
these
pathways
may
be
an
optimal
strategy
for
treating
can-‐
cers
of
this
subtype.
Acknowledgements:
Financial
support
was
provided
by
the
National
Cancer
Institute
(U01CA182740)
to
SLM
and
RHC.
AJR
was
supported
in
part
by
training
grant
R25GM095480.
References:
[1] Lehmann
BD,
Bauer
JA,
Chen
X,
Sanders
ME,
Chakravarthy
AB,
Shyr
Y,
Pietenpol
JA.
Identification
of
human
triple-‐negative
breast
cancer
subtypes
and
preclinical
models
for
selection
of
targeted
therapies.
J
Clin
Invest
2011;
121:
2750-‐2767
P478
Triterpene
saponin
constituents
from
roots
of
Bupleurum
falca-‐
tum:
Hepatoprotective
effects
on
D-‐galactosamine-‐induced
cell
damage
Takuya
Konno1,
Kiyofumi
Ninomiya1,
Masayuki
Yoshikawa2,
Hisashi
Matsuda2,
Toshio
Mori-‐
kawa1
1
Pharmaceutical
Research
and
Technology
Institute,
Kindai
University,
3-‐4-‐1
Kowakae,
Higashi-‐osaka,
Osaka,
577-‐8502,
Japan,
2
Kyoto
Pharmaceutical
University,
Misasagi,
Yamashina-‐ku,
Kyoto,
607-‐8412,
Japan
Bupleurum
falcatum
L.
[Apiaceae]
is
cultivated
in
Asia,
and
its
root
part,
Bupleuri
Radix
(“Sai-‐
ko"
in
Japanese),
is
one
of
the
most
important
natural
medicines
in
Japan.
In
the
Japanese
Pharmacopoeia
XVI,
the
root
has
been
used
for
anti-‐inflammatory,
anti-‐pyretic,
and
anti-‐
hepatotoxic
effects
in
the
treatments
of
common
cold,
fever,
and
hepatitis
[1].
We
have
found
that
MeOH
extract
of
the
roots
of
B.
falcatum
showed
inhibitory
activity
in
D-‐galactisamine
(D-‐
GalN)-‐induced
cell
damage
in
hepatocytes.
Through
bioassay-‐guided
separation,
several
triterpene
saponins
were
identified
as
the
bioactive
constituents.
Furthermore,
hepatoprotec-‐
tive
effects
of
major
active
saponins
against
D-‐GalN/lipopolysaccharide
(LPS)-‐induced
liver
injury
in
mice
were
also
examined.
From
the
MeOH
extract
of
the
roots
of
B.
falcatum
cultivat-‐
ed
in
Sichuan
province,
China,
a
new
and
17
known
saponins
were
isolated.
Hepatocytes
were
isolated
from
Wister
rat
by
a
collagenase
perfusion
method.
The
cell
suspension
at
4
×104
cells
in
100
μl
William's
E
medium
(containing
10%
FBS)
was
inoculated
in
a
96-‐well
micro-‐
plate
and
pre-‐incubated
for
4
h
at
37°C
under
5%
CO2
atmosphere.
After
pre-‐incubation,
100
μl
of
fresh
medium
containing
D-‐GalN
(1
mM)
with
the
test
sample
was
added
to
the
medium.
After
44
h
incubation,
cell
viability
was
assessed
by
MTT
colorimetric
assay.
The
structure
of
a
new
saponin
[1]
was
determined
on
the
basis
of
spectroscopic
properties
and
chemical
evi-‐
dence.
Among
the
isolates,
saikosaponins
b3
and
b4,
and
bupleuroside
IX
significantly
inhibit-‐
ed
cell
damage
in
hepatocytes.
In
addition,
major
saikosaponins,
such
as
saikosaponins
a,
c,
and
d,
were
found
to
significantly
inhibit
the
liver
injury.
The
data
demonstrated
that
saponin
constituents
from
B.
falcatum
roots
could
protect
against
D-‐GalN
and
D-‐GalN/LPS-‐induced
liver
injury.
H
CH2OH
HO OH
OO
OH O
O
HO H OH
OH O H
HO O OH
H
1
HO OH
Keywords:
Bupleurum
falcatum,
D-‐galactosamine,
D-‐GalN/LPS,
liver
failure
Reference:
[1] Nakahara
Y,
Okawa
M,
Kinjo
J,
Nohara
T.
Oleanene
glycosides
of
the
aerial
parts
and
seeds
of
Bupleurum
falcatum
and
the
aerial
parts
of
Bupleurum
rotundifolium,
and
their
evalua-‐
tion
as
anti-‐hepatitis
agents.
Chem
Pharm
Bull
2011;
59:
1329-‐1339
P479
Multidrug-‐efflux
inhibitors
–
A
study
on
selected
medicinal
plants
Tariq
M.
Aljarba,
Paul
Stapleton,
Simon
Gibbons
Research
Department
of
Pharmaceutical
and
Biological
Chemistry,
UCL
School
of
Pharmacy,
29-‐39
Brunswick
Square,
London
WC1N
1AX,
United
Kingdom
Multidrug-‐resistance
among
bacteria
has
become
a
global
issue
and
bacterial
resistance
oc-‐
curs
as
a
result
of
mutations
in
bacterial
genes
or
the
acquisition
of
resistance
determinants
borne
on
plasmids,
bacteriophages,
transposons
and/or
by
the
action
of
multidrug-‐efflux
pumps
[1].
These
pumps
work
to
remove
a
variation
of
structurally
unrelated
antibiotics
from
the
microorganism
resulting
in
reduced
susceptibility
of
the
antibiotic
[2].
This
study
has
evaluated
natural
products
for
their
in
vitro
antibiotic
potentiation
against
S.
aureus
strains
possessing
distinct
efflux-‐related
multidrug-‐resistance
pumps.
Quercus
robur,
Fraxinus
excel-‐
sior,
Fagus
sylvatica,
Ulmus
minor,
Robinia
pseudoacacia,
Baptisia
tinctoria
and
Frangula
alnus
were
extracted.
Hexane,
chloroform,
methanol
and
aqueous
extracts
and
fractions
of
each
of
the
samples
were
investigated
for
their
antibiotic-‐potentiation
activity
against
effluxing
strains
of
S.
aureus
using
a
modulation
assay.
These
strains
included
SA1199B
(fluoroquino-‐
lone-‐resistant;
NorA),
RN4220
(macrolide-‐resistant;
MsrA)
and
XU212
(tetracycline-‐resistant;
TetK).
Extracts
and
fractions
were
produced
using
a
Soxhlet
aparatus,
ultrasound-‐assisted.
The
results
showed
that
the
chloroform
extract
of
B.
tinctoria
and
R.
pseudoacacia
had
the
highest
antibiotic-‐potentiation
activity
against
all
S.
aureus
effluxing
strains.
They
also
en-‐
hanced
the
activity
of
norfloxacin
against
SA1199B
with
a
256-‐fold
reduction
in
norfloxacin
minimum
inhibitory
concentration.
The
chloroform
extract
of
B.
tinctoria
enhanced
the
activi-‐
ty
of
erythromycin
against
RN4220
with
a
128-‐fold
reduction.
The
potentiation
activity
of
these
extracts
indicates
that
efflux
inhibitor
natural
products
are
present
and
studies
are
un-‐
derway
to
isolate
and
characterise
these
compounds.
Keywords: Quercus
robur,
Fraxinus
excelsior,
Fagus
sylvatica,
Ulmus
minor,
Robinia
pseu-‐
doacacia,
Baptisia
tinctoria,
Frangula
alnus.
References:
[1] Piddock
LJV.
Clinically
relevant
chromosomally
encoded
multidrug
resistance
efflux
pumps
in
bacteria.
Clin
Microbiol
Rev
2006;
19:
382-‐402
[2] Smith
ECJ,
Williamson
EM,
Wareham
N,
Kaatz
GW,
Gibbons
S.
Antibacterials
and
modulators
of
bacterial
resistance
from
the
immature
cones
of
Chamaecyparis
lawsoniana.
Phytochemistry
2007;
68:
210-‐21
P480
Selected
medicinal
plant
extracts
as
inhibitors
of
conjugal
transfer
of
plasmid-‐mediated
antibiotic
resistance
in
Escherichia
coli
Tariq M. Aljarba, Paul Stapleton, Simon Gibbons
Research
Department
of
Pharmaceutical
and
Biological
Chemistry,
UCL
School
of
Pharmacy,
29-‐39
Brunswick
Square,
London
WC1N
1AX,
United
Kingdom
Antibiotic
resistance
is
a
global
health
concern
and
plasmids
are
a
major
mechanism
of
spreading
bacterial
resistance.
Plasmids
transfer
the
genes
responsible
for
the
resistance
by
four
secretion
steps.
Inhibiting
this
process
is
potentially
a
good
therapeutic
target
identifying
substances
modulating
antibiotic
resistance
[1].
In
this
study,
selected
medicinal
plant
ex-‐
tracts
were
screened
for
their
ability
to
inhibit
the
conjugal
transfer
of
plasmids
in
Escherichia
coli.
Bacteria
were
transformed
with
a
plasmid
containing
either
the
drug
resistance
gene
in
E.
coli
with
the
PKM
101
plasmid
(encoding
for
ampicillin
resistance),
or
the
TP114
plasmid
(kanamycin
resistance).
The
donor
for
the
bacterial
conjugation
assay
was
E.
coli
possessing
the
ER
1793
plasmid,
which
encodes
for
streptomycin
resistance
[2].
The
test
samples
used
were
extracts
and
fractions
from
Quercus
robur,
Robinia
pseudoacacia
and
Baptisia
tinctoria.
The
methanol
extract
of
Quercus
robur
showed
a
strong
reduction
in
the
conjugal
transfer
of
plasmid
PKM101
with
a
percentage
transfer
frequency
of
plasmids
to
be
4%
in
each
case
and
below
when
compared
to
the
control.
The
percentage
transfer
frequencies
for
Quercus
robur
hexane
extract
were
found
to
be
greater
than
50%,
indicating
that
they
were
poorly
active
against
conjugal
transfer
of
TP114.
This
could
mean
that
other
plant
extracts
such
as
Quercus
robur
methanol
and
Quercus
robur
hexane
extract
may
increase
the
transfer
frequency
of
TP114.
Possibly
the
conjugal
inhibitory
activity
of
these
natural
products
is
due
to
the
inhibi-‐
tion
of
conjugative
pili
formation.
Further
studies
are
ongoing
to
isolate
and
characterise
the
natural
products
responsible.
Keywords:
Plasmids,
conjugation,
Quercus
robur,
Robinia
pseudoacacia,
Baptisia
tinctoria.
References:
[1] Fernandez-‐Lopez
R,
Machón
C,
Longshaw
CM,
Martin
S,
Molin
S,
Zechner
EL,
Espinosa
M,
Lanka
E,
de
la
Cruz
F.
Unsaturated
fatty
acids
are
inhibitors
of
bacterial
conjugation.
Microbiology,
2005;
151:
3517-‐3526
[2] Livermore,
D.M.
Bacterial
resistance:
origins,
epidemiology,
and
impact.
Clinical
infectious
diseases :
an
official
publication
of
the
Infectious
Diseases
Society
of
America,
2003;
36(Suppl
1):
S11-‐S23
P481
A
new
strategy
for
the
evaluation
of
biological
properties
of
lipo-‐
philic
natural
products:
Application
to
the
wound
healing
and
an-‐
tibacterial
activities
of
Calophyllum
inophyllum
Linn
oil
Teddy Léguillier1, Marylin Lecsö-‐Bornet2, Christelle Lémus3, Nicolas Lebouvier4, Edouard
Hnawia4, Mohammed Nour4, William Aalbersberg5, Phila Raharivelomanana6, Patrice Rat1
1 Department of Analytical and Cellular Toxicology CNRS UMR 8638, Faculté de Pharmacie de Paris,
75006
Paris,
France;
2
Department
of
Intestinal
Ecosystem,
Probiotics
and
Antibiotics
EA
4065,
Faculté
de
Pharmacie
de
Paris,
75006
Paris,
France;
3
Department
of
Environment
and
Natural
Resources
AIHP
GE-‐
ODE
EA
929,
Université
des
Antilles,
97233
Martinique,
France;
4
Department
of
Insular
Environment
and
Natural
Resources
LIVE-‐EA
4243,
Université
de
la
Nouvelle-‐Calédonie,
98851
Nouméa,
Nouvelle
Calédonie;
5
Institute
of
Applied
Sciences,
The
University
of
the
South
Pacific,
Laucala
Campus,
Suva,
Fiji.
6
Department
of
Oceanian
Insular
Ecosystem-‐EIMS
UMR
241,
Université
de
la
Polynésie
Française,
Tahiti,
98702
FAA'A,
Polynésie
Française.
Calophyllum
inophyllum
Linn
(Calophyllaceae)
is
an
evergreen
tree
ethno-‐medically
used
across
Oceania,
especially
in
Polynesia
[1].
Bark,
leaves
and
seeds
are
used
for
their
therapeu-‐
tic
virtues.
Particularly,
the
oil
extracted
from
Calophyllum
inophyllum
seeds
is
traditionally
used
topically
to
treat
a
wide
range
of
skin
injuries
including
burn,
scar,
infected
wounds,
and
also
skin
diseases
such
as
dermatosis,
urticaria
and
eczema
[2,3].
Recently,
various
publica-‐
tions
have
documented
the
therapeutic
worth
of
medicinal
plants
to
validate
the
empirical
claims
of
their
biological
activity.
However,
very
few
scientific
studies
reported
the
therapeu-‐
tic
properties
of
Calophyllum
inophyllum
oil
(CIO).
Indeed,
modern
techniques
used
to
investi-‐
gate
the
biological
properties
of
natural
products
are
mostly
designed
for
the
evaluation
of
hydrophilic
products.
As
a
consequence,
when
natural
lipophilic
products
are
evaluated,
or-‐
ganic
solvents
are
used
and
could
interfere
with
the
expected
result
[4].
Here,
we
developed
a
new
strategy
convenient
to
assess
biological
properties
of
lipophilic
natural
products
without
the
use
of
any
organic
solvent.
Using
the
Microplate
Alamar
Blue
Assay
(MABA),
scratch
test,
oilogram
and
bioautography
assay,
we
investigated
CIO
biological
properties.
Based
on
this
new
strategy,
we
confirmed
the
pharmacological
effects
of
CIO
from
Indonesia,
Tahiti,
Fiji
Is-‐
lands
and
New
Caledonia
as
a
potent
wound
healing
and
antimicrobial
agent
[5].
Finally,
the
successful
use
of
this
innovative
strategy
could
be
extended
to
other
natural
lipophilic
prod-‐
ucts
such
as
vegetable
and
essential
oils.
Acknowledgements:
We
thank
F.
Duveau,
M.
Dutot,
E.
Olivier,
A.
Wakx
for
technical
advice
and
helpful
discussions.
We
are
grateful
to
J.
Bennett,
J-‐C
Bobbia,
J-‐L.
Delubriat,
A.
Rannou
and
O.
Touboul
for
provid-‐
ing
Calophyllum
inophyllum
oils.
This
work
was
supported
by
Adebiopharm
ER67
and
by
the
Fonds
Pacifique
(CALO-‐PS
project).
Keywords:
Calophyllum
inophyllum
oil,
lipophilic
natural
product,
wound
healing,
antimicro-‐
bial
agent,
medicinal
plants.
References:
[1] Petard
P.
The
use
of
Polynesian
medicinal
plants
in
Tahitian
medicine.
Noumea,
New
Cal-‐
edonia:
South
Pacific
Commission;
1972,
66
pp.
[2] Chevalier
J.
Study
on
a
new
cicatrizing
agent
for
cutaneous
and
mucous
wounds,
oil
of
Calophyllum
inophyllum
(dissertation).
Paris:
Institut
de
Biologie
Normale
Superieure;
1951
[3] Pocidalo
JJ,
Chaslot
M.
Action
of
oil
of
Calophyllum
on
experimental
burns.
Comptes
Rendus
Séances
Société
Biol
Ses
Fil,
1955;
149:
357-‐359
[4] Eldin
HME,
Sarhan
RM.
Cytotoxic
effect
of
organic
solvents
and
surfactant
agents
on
Acan-‐
thamoeba
castellanii
cysts.
Parasitol
Res
2014;
113:
1949-‐1953
[5] Léguillier
T,
Lecsö-‐Bornet
M,
Lémus
C,
Rousseau-‐Ralliard
D,
Lebouvier
N,
Hnawia
E,
Nour
M,
Aalbersberg
W,
Ghazi
K,
Raharivelomanana
P,
Rat
P.
The
wound
healing
and
antibacterial
activity
of
five
ethnomedical
Calophyllum
inophyllum
Oils:
an
alternative
therapeutic
strategy
to
treat
infected
wounds.
PLOS
ONE
2015;
10:
e0138602
P482
Arginase
inhibitors:
from
chlorogenic
acid
to
cinnamides
Thanh-‐Nhat
Pham1,
Duc-‐Thu
Trinh2,
Simon
Bordage1,3,
Céline
Demougeot1,
Marc
Pudlo1,
Khac-‐Minh
Thai2,
Corine
Girard-‐Thernier1
1FDE
EA4267,
University
of
Bourgogne
Franche-‐Comté,
F-‐25000
Besançon,
France;
2Department
of
Me-‐
dicinal
Chemistry,
Faculty
of
Pharmacy,
University
of
Medecine
and
Pharmacy
at
Ho
Chi
Minh
City,
41
Dinh
Tien
Hoang,
Dist.
1,
Ho
Chi
Minh
City,
Vietnam;
3EA
7394
-‐
ICV
-‐
Institut
Charles
Viollette,
University
of
Lille,
F-‐59000
Lille,
France
The
interest
to
consider
arginase,
a
metalloenzyme
hydrolysing
L-‐arginine
to
L-‐ornithine
and
urea,
as
a
therapeutic
target
has
grown
in
the
recent
years
and
the
use
of
arginase
inhibitors
has
been
proved
to
be
beneficial
in
cardiovascular
and
nervous
system
diseases
[1].
However,
the
most
potent
commercial
inhibitors
cannot
be
used
in
clinic
due
to
their
pharmacokinetic
and
toxicological
properties
[2].
Search
for
new
inhibitors
is
thus
required
and
such
com-‐
pounds
may
be
inspired
by
natural
substances
[3].
Further
to
a
screening
of
a
series
of
natural
polyphenols
on
an
arginase
inhibitory
assay
using
purified
liver
bovine
arginase
(b-‐ARG
I),
we
showed
that
chlorogenic
acid
could
serve
as
a
lead
compound
to
design
new
arginase
inhibi-‐
tors
[4].
In
particular,
our
findings
suggested
that
the
caffeoyl
moiety
could
be
crucial
for
ar-‐
ginase
inhibition.
In
the
present
study,
we
focused
on
development
of
chlorogenic
acid
deriva-‐
tives
wherein
the
quinoyl
moiety
was
replaced
by
a
phenethylamine.
A
series
of
cinnamide
derivatives
was
thus
synthesized
and
evaluated
for
their
in
vitro
inhibitory
activity
on
b-‐ARG
I.
Moreover,
a
homology
structure
of
b-‐ARG
I
was
created
and
used
for
molecular
docking
to
predict
the
interaction
of
cinnamides
toward
the
active
site
of
the
enzyme.
Our
results
showed
that
caffeic
acid
phenethyl
amide
(CAPA)
was
the
most
potent
inhibitor
(IC50
value
6.9
±
1.3
µM),
still
weaker
than
the
reference
inhibitor
nor-‐NOHA
(IC50
value
1.7
±
0.2
µM)
but
slightly
more
active
than
chlorogenic
acid
(IC50
value
10.6
±
0.8
µM).
Structure-‐activity
rela-‐
tionship
showed
that
cinnamoyl
moiety
and
catechol
function
were
important
for
inhibitory
activity.
Docking
results
demonstrated
that
caffeoyl
moiety
could
penetrate
into
the
active-‐
site
pocket
whereas
catechol
and
amide
groups
might
interact
with
several
crucial
residues
involved
in
the
hydrolysis
mechanism
of
enzyme.
This
study
suggests
that
3,4-‐
dihydroxycinnamides
could
be
considered
as
potential
arginase
inhibitors.
Acknowledgements:
The
Ministère
Français
de
l’Enseignement
supérieur
et
de
la
Recherche
for
awarding
a
PhD
fellowship
to
T.-‐N.
Pham.
The
molecular
modeling
work
was
supported
by
the
Vietnam's
National
Foundation
for
Science
and
Technology
Development
-‐
NAFOSTED
(Grant
#
106-‐YS.05-‐2015.31
to
Khac-‐
Minh
Thai).
Andy
Zedet
is
acknowledged
for
technical
assistance.
Keywords: liver bovine arginase, inhibitors, polyphenols, docking study, cinnamides.
References:
[1] Caldwell
RB,
Toque
HA,
Narayanan
SP,
Caldwell
RW.
Arginase:
an
old
enzyme
with
new
tricks.
Trends
Pharmacol
Sci
2015;
36:
395−405
[2] Ivanenkov
YA,
Chufarova
NV.
Small-‐molecule
arginase
inhibitors.
Pharm
Pat
Anal
2013;
3:
65–85
[3] Girard-‐Thernier
C,
Pham
TN,
Demougeot
C.
The
promise
of
plant-‐derived
substances
as
inhibitors
of
arginase.
Mini
Rev
Med
Chem
2015;
15:
798−808
[4] Pham
TN,
Guglielmetti
AS,
Fimbel
S,
Demougeot
C,
Girard-‐Thernier
C.
Arginase
inhibitory
activity
of
several
natural
polyphenols
using
a
novel
in
vitro
test
on
purified
arginase.
Planta
Med
2014;
80:
P1L9
P483
Anticonvulsant
agents
from
Boswellia
sacra
identified
by
zebrafish
bioassay-‐guided
fractionation
Théo
Brillatz1,
Emerson
Ferreira
Queiroz1,
Laurence
Marcourt1,
Maxime
Jacmin2,
Alexander
D.
Crawford2,
Jean-‐Luc
Wolfender1
1School
of
Pharmaceutical
Sciences,
University
of
Geneva,
University
of
Lausanne,
Quai
Ernest-‐Ansermet
30,
CH-‐1211
Geneva
4,
Switzerland,
2Luxembourg
Center
for
Systems
Biomedicine,
Université
du
Luxem-‐
bourg,
6
avenue
du
swing,
L-‐4367
Belvaux,
Luxembourg
Epilepsy
is
a
chronic
CNS
disorder
characterised
by
recurrent
seizures.
It
is
the
most
preva-‐
lent
neurological
disease
worldwide,
with
over
70
million
people
afflicted
[1].
Despite
the
availability
of
over
30
approved
anti-‐epileptic
drugs
(AEDs),
a
third
of
treated
epilepsy
pa-‐
tients
experience
serious
side
effects
to
these
medications,
and
another
third
do
not
respond
at
all
and
are
referred
to
as
being
“pharmacoresistant”.
Thus,
there
is
a
clear
need
for
new
AEDs.
Towards
this
end,
we
investigated
the
sacred
frankincense
from
Boswellia
sacra
Flueck.
[Burseraceae],
which
has
been
used
as
a
medicinal
drug
in
many
parts
of
the
world
for
thou-‐
sands
of
years.
The
hexane
extract
of
the
resin
of
this
plant
exhibited
significant
anticonvul-‐
sant
activity
in
zebrafish
larvae
with
seizures
induced
by
the
GABAA
antagonist
pentylene-‐
tetrazol
(PTZ)
[2].
In
order
to
easily
isolate
and
identify
the
active
compounds,
the
analytical
HPLC-‐PDA-‐ELSD
conditions
were
transferred
geometrically
to
a
preparative
medium-‐
pressure
liquid
chromatography
column
(MPLC-‐UV-‐ELSD)
using
chromatographic
calcula-‐
tions
[3].
This
gradient
transfer
ensured
that
the
same
selectivity
and
elution
order
was
kept
between
the
analytical
and
the
preparative
scale
and
provided
an
efficient
isolation
of
the
ac-‐
tive
compounds
at
the
milligram
scale.
The
hexane
extract
was
fractionated
in
33
fractions
and
eight
major
compounds
were
isolated
and
characterized
as
terpenes,
including
a
new
diterpene.
Incensole
and
β-‐boswellic
acid
showed
significant
in
vivo
anticonvulsant
activity
and
decreased
respectively
31
%
(10
µg/ml)
and
90
%
(100
µg/ml)
of
the
induced
sei-‐
zures.
The
present
work
reports
for
the
first
time
the
anticonvulsant
activity
of
isolated
com-‐
pounds
from
B.
sacra
and
confirmed
the
anti-‐epileptic
activity
of
Boswellia
species,
validating
their
use
in
traditional
medicine
to
treat
epileptic
seizures.
Keywords: Ethnopharmacology, Boswellia sacra, epilepsy, anticonvulsant, zebrafish
References:
[1]
Ngugi
AK,
Bottomley
C,
Kleinschmidt
I,
Sander
JW,
Newton
CR.
Estimation
of
the
burden
of
active
and
life-‐time
epilepsy:
a
meta-‐analytic
approach.
Epilepsia
2010;
51:
883−890
[2]
Challal
S,
Buenafe
OEM,
Queiroz
EF,
Maljevic
S,
Marcourt
L,
Bock
M,
Kloeti
W,
Dayrit
FM,
Harvey
AL,
Lerche
H,
Esguerra
CV,
Witte
PAM,
Wolfender
JL,
Crawford
AD.
Anticonvulsant
steroid
glycosides
from
the
Philippine
medicinal
plant
Solanum
torvum.
ACS
Chem
Neuro-‐
sci
2015;
15:
993−1004
[3]
Challal
S,
Queiroz
EF,
Debrus
B,
Kloeti
W,
Guillarme
D,
Gupta
MP,
Wolfender
JL.
Rational
and
efficient
preparative
isolation
of
natural
products
by
MPLC-‐UV-‐ELSD
based
on
HPLC
to
MPLC
gradient
transfer.
Planta
Med
2015;
81:
1636−1643
P484
Bioguided
isolation
of
anticonvulsant
principles
from
Helleborus
cyclophyllus
using
the
zebrafish
epilepsy
model
Théo Brillatz1, Emerson Ferreira Queiroz1, Laurence Marcourt1, Konstantina Vougogi-‐
annopoulou2, Maxime Jacmin3, Alexander D. Crawford3, Leandros Skaltsounis2, Jean-‐Luc
Wolfender1
1School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Quai Ernest-‐Ansermet
30,
CH-‐1211
Geneva
4,
Switzerland,
2Department
of
Pharmacognosy
&
Natural
Product
Chemistry,
School
of
Pharmacy,
University
of
Athens,
Panepistimioupolis,
Zografou,
15771
Athens,
Greece,
3Luxembourg
Center
for
Systems
Biomedicine,
Université
du
Luxembourg,
6
avenue
du
swing,
L-‐4367
Belvaux,
Luxem-‐
bourg
In
ancient
Greece,
certain
Helleborus
species
were
used
to
treat
a
variety
of
diseases
including
leprosy,
deafness,
madness
and
epilepsy
[1].
Hippocrates
(~400
BC)
discovered
diuretic
and
emetic
properties
of
the
Greek
Hellebore
Helleborus
cyclophyllus
Boissier
[Ranunculaceae]
while
in
ancient
Rome
this
plant
was
recommended
for
curing
epilepsy,
madness
and
melan-‐
cholia
[2].
Based
on
our
ethnopharmacological
survey,
we
hypothesized
that
H.
cyclophyllus
might
reveal
novel
anticonvulsant
natural
products.
Towards
this
end,
leaves
and
root
ex-‐
tracts
of
this
plant
were
screened
in
a
zebrafish
epilepsy
model
with
seizures
induced
by
the
GABAA
antagonist
pentylenetetrazol
[3].
As
expected,
the
methanolic
root
extract
showed
a
significant
inhibition
of
seizures.
In
order
to
target
the
bioactivity
of
the
active
extract,
the
HPLC
analytical
conditions
were
geometrically
transferred
for
a
high-‐scale
fractionation
on
medium-‐pressure
liquid
chromatography
(MPLC-‐UV-‐ELSD).
Major
compounds
from
these
fractions
were
isolated
by
semi-‐preparative
chromatography
and
identified
by
HRMS
and
NMR,
including
a
new
saponin.
Subsequently,
three
out
of
sixteen
fractions
exhibited
potent
anticonvulsant
activity.
Finally,
hellebrin
and
two
other
saponins
showed
significant
reduc-‐
tion
of
epileptic
seizures
on
the
zebrafish
bioassay.
Our
results
report
for
the
first
time
the
anticonvulsant
activity
of
some
isolated
compounds
from
Helleborus
cyclophyllus
and
con-‐
firmed
one
of
the
medicinal
use
of
this
ancient
Greek
plant.
Keywords: Ethnopharmacology, Helleborus cyclophyllus, epilepsy, anticonvulsant, zebrafish
References:
[1] Jäger
AK,
Gauguin
B,
Adsersen
A,
Gudiksen
L.
Screening
of
plants
used
in
Danish
folk
medi-‐
cine
to
treat
epilepsy
and
convulsions.
J
Ethnopharmacol
2006;
105:
294−300
[2] Ustinova
Y,
Cardeña
E.
Combat
stress
disorders
and
their
treatment
in
ancient
Greece.
Psychol
Trauma
2014;
6:
739−748
[3] Challal
S,
Buenafe
OEM,
Queiroz
EF,
Maljevic
S,
Marcourt
L,
Bock
M,
Kloeti
W,
Dayrit
FM,
Harvey
AL,
Lerche
H,
Esguerra
CV,
Witte
PAM,
Wolfender
JL,
Crawford
AD.
Anticonvul-‐
sant
steroid
glycosides
from
the
Philippine
medicinal
plant
Solanum
torvum.
ACS
Chem
Neurosci
2015;
15:
993−1004
P485
New
compounds
from
the
leaves
of
Cleistocalyx
operculatus
and
their
inhibitory
activities
on
influenza
A
neuraminidases
Thi-‐Kim-‐Quy
Ha1,
Won
Keun
Oh1*
1Korea
Bioactive
Natural
Material
Bank,
Research
Institute
of
Pharmaceutical
Sciences,
College
of
Phar-‐
macy, Seoul National University, Seoul 08826, Republic of Korea.
Medicinal plant extracts from the natural material extraction library were screened for their
effects on anti-‐influenza virus activities. During this process, the ethanolic extract of Cleistoca-‐
lyx operculatus leaves was found to exhibit potential neuraminidase (NA) inhibitory activity.
C. operculatus (Roxb.) Merr. and Perry (Myrtaceae) has been used in Vietnamese folk medi-‐
cine since ancient times. Its buds and leaves were traditionally used to treat fever from the
common cold, bacillary dysentery and gastric inflammation. Previous reports have demon-‐
strated that flavonoids are the main chemical constituents of the buds [1], with anti-‐oxidant
[2], anti-‐hyperglycemic [3], anti-‐influenza [1], and cholinesterase inhibitory activities [4]. Bio-‐
assay-‐guided fractionation of C. operculatus led to the isolation of two new acetophenones (1
and 2) and one new flavanone (3), along with six known compounds (4-‐9). The structures of
all isolated compounds were elucidated using extensive spectroscopic methods and through
comparison with the data of previous literatures. Compounds 6 and 8 exhibited strong enzy-‐
matic inhibitions on various neuraminidases from different influenza viruses, including H1N1,
H9N2, novel H1N1, and oseltamivir-‐resistant novel H1N1 (H274Y mutation) expressed in
HEK293 cells (IC50 values ranging from 5.07 ± 0.94 µM to 9.34 ± 2.52 µM, respectively). In ad-‐
dition, kinetic analysis revealed that these compounds inhibited the H1N1 neuraminidase en-‐
zyme with the non-‐competitive mode. Compounds 6 and 8 were also assessed for their inhibi-‐
tory effects on viral replication using a cytopathic effect (CPE) reduction assay of swine influ-‐
enza H1N1 A/PR/8/34 in MDCK cells. Furthermore, the presence of large amounts of these
major compounds (6 and 8) suggested their possibility to be exploited for further work on the
synthesis of analogues for the development of novel NA inhibitors in the future.
Keywords:
Cleistocalyx
operculatus,
Myrtaceae,
H1N1,
influenza,
neuraminidase.
References:
[1] Dao
TT,
Tung
BT,
Nguyen
PH,
Thuong
PT,
Yoo
SS,
Kim
EH,
Kim
SK,
Oh
WK.
C-‐Methylated
flavonoids
from
Cleistocalyx
operculatus
and
their
inhibitory
effects
on
novel
influenza
A
(H1N1)
neuraminidase.
J
Nat
Prod
2010;
73:
1636-‐1642
[2] Min
BS,
Thu
CV,
Dat
NT,
Dang
NH,
Jang
HS,
Hung
TM.
Antioxidative
flavonoids
from
Cleis-‐
tocalyx
operculatus
buds.
Chem
Pharm
Bull
2008;
56:
1725-‐1728
[3] Mai
TT,
Chuyen
NV.
Anti-‐hyperglycemic
activity
of
an
aqueous
extract
from
flower
buds
of
Cleistocalyx
operculatus
(Roxb.)
Merr
and
Perry.
Biosci
Biotech
Biochem
2007;
71:
69-‐76
[4] Min
BS,
Cuong
TD,
Lee
JS,
Shin
BS,
Woo
MH,
Hung
TM.
Cholinesterase
inhibitors
from
Cleistocalyx
operculatus
buds.
Arch
Pharm
Res
2010;
33:
1665-‐1670
P486
Antifungal
activities
and
chemical
composition
of
the
essential
oil
of
Lippia
micromera
(Verbenaceae)
cultivated
in
French
Guiana
Camille
Scotto1,
Pauline
Burger1,
Thomas
Michel1,
Mehdi
Khodjet
el
khil2,
Marine
Ginouves3,
Ghislaine
Prevot3,
Denis
Blanchet3,4,
Piero
G.
Delprete5,
Xavier
Fernandez1
1
Institut
de
Chimie
de
Nice,
UMR
7272,
Parc
Valrose,
06108
Nice
Cedex
2,
France,
2
Guyarômes,
Route
Nationale
2,
PK
6.5,
97351
Matoury,
Cayenne,
French
Guiana,
France,
3
Université
de
la
Guyane,
Labora-‐
toire
d’épidémiologie
des
parasitoses
tropicales
EA
3593
–
Labex
CEBA
UFR
de
médecine,
Cayenne,
French
Guiana,
France,
4
Laboratoire
hospitalo-‐universitaire
de
parasitologie
et
mycologie,
Centre
hospi-‐
talier
de
Cayenne,
Cayenne,
French
Guiana,
France,
5
Herbier
IRD
de
Guyane,
Institut
de
Recherche
pour
le
Développement
(IRD),
UMR
AMAP,
Boite
Postale
165,
97323
Cayenne
Cedex,
French
Guiana,
France
French
Guyana
is
a
part
of
the
Amazonia
rainforest
identified
as
a
biodiversity
hotspot
for
both
animal
and
plant
species
threatened
with
extinction
[1].
With
the
generalized
awareness
of
this
endangered
richness
a
number
of
conservation
and
promotion
actions
have
been
un-‐
dertaken
over
the
last
years.
In
collaboration
with
the
Guyarômes
society,
the
potential
valori-‐
zation
of
essential
oils
from
local
plants
on
the
international
market
is
assessed.
Lippia
mi-‐
cromera
Schauer
(Verbenaceae
family)
is
a
strong
aromatic
shrub
with
many
branched
and
white
tiny
flowers
with
yellow
throat.
Its
origin
is
not
well
established
but
it
is
assumed
that
the
plant
is
from
northern
South
America
(Colombia,
Venezuela
and
Guiana),
the
Caribbean
and
Nicaragua.
[2]
It
is
one
of
the
selected
plants
for
such
a
valorization
action.
The
species
is
well
known
for
culinary
seasoning,
anti-‐inflamatory,
anti-‐microbial
and
antioxidant
activities
[3].
The
present
poster
reports
for
the
first
time
the
analysis
of
the
chemical
composition
by
GC-‐FID
and
GC-‐MS
of
the
essential
oil
of
L.
micromera
organically
cultivated
in
French
Guiana.
A
total
of
thirty-‐one
components
accounting
for
97%
of
the
total
GC-‐FID
chromatogram
were
identified.
The
evaluation
of
its
antimicrobial
and
antiparasitic
activities
against
strains
of
several
Candida
spp.
(MIC)
and
Leishmania
guyanensis
(IC50)
respectively
shows
a
strong
an-‐
timicrobial
activity
ranging
from
125
μg/ml
to
500
μg/ml
according
to
the
Candida
species
tested,
while
the
anti-‐leishmania
activity
seems
negligible.
Thus
the
EO
was
assessed
for
its
anti-‐ageing
and
antioxidant
properties
(in
vitro
tests
in
96-‐wells
plates)
and
revealed
also
a
strong
percentage
of
inhibition
(more
than
60%)
for
elastase
and
lipoxygenase
assays.
Acknowledgements:
The
authors
are
grateful
to
Guyarômes
for
the
financial
support
they
provided
and
for
making
the
essential
oil
samples
available
Keywords:
Lippia
micromera,
Verbenaceae,
French
Guiana,
antifungal
and
antiparasitic
activ-‐
ities,
Candida
spp,
Leishmania
guyanensis
References:
[1] Myers
N,
Mittermeier
RA,
Mittermeier
CG,
da
Fonseca
GAB,
Ken
J.
Biodiversity
hotspots
for
conservation
priorities.
Nature
2000;
403:
853-‐858
[2] Staples
GW,
Kristiansen
MS,
Lippia
micromera
Schauer
Verbenaceae,
the
verbena
family
In:
Ethnic
culinary
herbs:
A
guide
to
identification
and
cultivation
in
Hawaii.
Edits.,
Uni-‐
versity
of
Hawaii
Press
1999;
Honolulu:
56-‐57
[3] Zapata
B,
Betancur-‐Galvis
L,
Duran
C,
Stashenko
E,
Cytotoxic
activity
of
Asteraceae
and
Verbenaceae
family
essential
oils.
J
Essent
Oil
Res
2014;
26:
50-‐57
P487
Anti-‐inflammatory,
cytotoxic
and
antimicrobial
activities
of
Piper
peltatum
leaf
extract
Thomas
Michel1,
Audrey
Kerdudo1,
Emy
Njoh
Ellong2,
Vanessa
Gonnot1,
Stéphane
Rocchi3,4,
Jean-‐François
Tanti3,4,
Laurent
Boyer3,4,
Sandra
Adenet2,
Katia
Rochefort2,
Xavier
Fernandez1
1Institut
de
Chimie
de
Nice,
UMR
CNRS
7272,
Université
Nice
Sophia
Antipolis,
Parc
Valrose,
06108
Nice
CEDEX
2,
France,
2Pôle
Agroalimentaire
Régional
de
Martinique
(P.A.R.M.),
Impasse
Petit
Morne
n°375,
97232
Lamentin,
Martinique,
France,
3INSERM,
U1065,
Centre
Méditerranéen
de
médecine
Moléculaire,
C3M,
Toxines
Microbiennes
dans
la
relation
hôte
pathogènes,
Nice,
France,
4Université
de
Nice-‐Sophia-‐
Antipolis,
UFR
Médecine,
Nice,
France
Piper
peltatum
L.
(syn.
Lepianthes
peltata
(L.)
Raf.
Ex
R.A.
Howard)
is
a
plant
belonging
to
the
Piperaceae
family
widely
distributed
in
the
Caribbean
and
in
tropical
America
regions.
Leaves
of
P.
peltatum
are
widely
used
in
traditional
medicine
of
tropical
Amazonian
(Peru,
Bolivia
and
Brazil)
and
Caribbean
regions
to
treat
inflammation,
fever,
hepatitis,
malaria
and
infec-‐
tious
diseases
[1].
In
current
study,
solvent
extracts
of
P.
peltatum
leaves
were
evaluated
for
their
anti-‐melanoma
(A375
metastatic
melanoma
cell
line),
anti-‐leukaemia
(K562
chronic
myeloid
leukemia
cell),
antimicrobial
activities
as
well
as
for
their
ability
to
block
activation
by
LPS
lipopolysaccharides
(LPS)
of
two
major
inflammatory
pathways
nuclear
factor-‐kappa
B
(NF-‐KB)
and
activator
protein
1
(AP1)
in
a
macrophage
lineage
[2].
The
results
show
that
cyclohexane
and
dichloromethane-‐methanol
(1:3,
v/v)
extracts
inhibit
both
NF-‐KB
and
AP1
transcription
factors
(Figure
1),
while
only
dichloromethane-‐methanol
extract
was
cytotoxic
against
A375
melanoma
and
K562
cells.
120
100
Phosphatase
alkaline
activity
80
60
40
20
0
-‐ + -‐ + -‐ + -‐ +
Control Cyclohexane DCM/MeOH MeOH/H2O
Figure
1:
Phosphatase
alkaline
activity
of
control
(without
extract),
cyclohexane,
dichloro-‐
methane/methanol
(DCM/MeOH,
1:3,
v/v),
and
hydroalcoolic
(H2O/MeOH,
1:1,
v/v)
extracts
of
L.
Peltatum
with
(+)
or
without
(-‐)
Antimicrobial
screening
of
the
crude
extracts
also
underline
the
dichloromethane-‐methanol
extract
as
the
most
active.
It
presents
a
high
antibacterial
activity
(85
%
of
inhibition)
against
resistant
strain
of
Staphylococcus
aureus
RN4220
and
a
slight
antibacterial
activity
against
S.
aureus
S25
at
12µg/mL.
Bioguided
fractionation
using
C18
solid
phase
extraction
cartridge
and
molecule
isolation
by
semi-‐preparative
liquid
chromatography
led
to
the
identification
of
three
active
metabolites:
4-‐nerolidylcatechol
(1)
and
two
compounds
for
the
first
time
de-‐
scribed
P.
pelatatum
gonzalitosin
I
(2)
and
sesamin
(3).
O O
O O O
HO O O
HO O O
OH O
O
(1)
4-‐nerolidylcatechol
(2)
Gonzalitosin
I
(3)
Sesamin
Keywords: Piper peltatum, anti-‐inflammatory, anti-‐melanoma, anti-‐leukaemia, anti-‐microbial,
bioguided fractionation
References:
[1] Lans
C,
Harper
T,
Georges
K,
Bridgewater
E.
Medicinal
and
ethnoveterinary
remedies
of
hunters
in
Trinidad.
BMC
Complement
Altern
Med
2001;
1:
10
[2] Tanti
JF,
Ceppo
F,
Jager
J,
Berthou
F.
Implication
of
inflammatory
signaling
pathways
in
obesity-‐induced
insulin
resistance.
Front
Endocrinol
(Lausanne)
2013;
3:
181
P488
Dragonbloodin
A1
and
A2:
novel
flavan
trimers
and
anti-‐
inflammatory
principles
from
Sanguis
Draconis
Wen-‐Ke
Du,
Hsin-‐Yi
Hung,
Ping-‐Chung
Kuo,
Tian-‐Shung
Wu
School
of
Pharmacy,
National
Cheng
Kung
University
Hospital,
College
of
Medicine,
National
Cheng
Kung
University,
701,
Tainan,
Taiwan
Inflammation
plays
a
key
role
in
many
disease
conditions.
Sanguis
Draconis,
also
known
as
dragon’s
blood,
is
a
deep
red
resin,
an
important
traditional
Chinese
medicine
(TCM)
for
blood
regulation
[1,
2].
The
pharmacological
studies
on
Daemonorops
draco
are
in
several
fields:
anti-‐coagulation,
anti-‐viral,
anti-‐bacterial,
anti-‐inflammatory,
anti-‐cancer
and
osteogenic
ac-‐
tivity
[3-‐8].
However,
studies
on
the
relationship
of
the
chemical
constituents
and
their
bioac-‐
tivities
are
rare,
which
drew
our
attention
to
investigating
novel
bioactive
compounds
from
this
important
TCM.
The
aim
of
the
work
was
to
isolate
novel
bioactive
compounds
from
dragon’s
blood
and
to
evaluate
their
anti-‐inflammatory
activity.
Two
flavan
trimers,
drag-‐
onbloodin
A1
(1)
and
A2
(2)
were
isolated
as
diastereomers.
The
structures
of
1
and
2
were
elucidated
by
spectroscopic
analysis
and
X-‐ray
diffraction.
Possible
biosynthesis
pathways
were
deduced.
IC50
values
of
dragonbloodin
A
for
inhibition
of
human
neutrophil
superoxide
anion
generation
and
elastase
release
were
>
10
µM
and
6.53
µM,
respectively.
In
addition,
inhibitions
of
neutrophil
elastase
release
of
dragonbloodin
A1
and
A2
were
seen
in
a
dose-‐
dependent
manner
from
1
µM
to
10
µM.
Figure
1.
Structures
of
dragonbloodin
A1
and
A2.
Acknowledgement:
The
authors
are
thankful
to
the
Ministry
of
Science
and
Technology
for
the
financial
support
of
the
present
research
Keywords:
Sanguis
Draconis,
traditional
Chinese
medicine,
X-‐ray,
human
neutrophil
elastase,
anti-‐inflammation
References:
[1] Pisoschi
AM,
Pop
A.
The
role
of
antioxidants
in
the
chemistry
of
oxidative
stress:
A
review.
Eur
J
Med
Chem
2015;
97:
55−74
[2] Chinese
Herbal
Medicine:
Materia
Medica
3rd
ed.
;Bensky,
D.
G.
K.,
T.
Eds,;
Eastland
Press
1993
[3] Gupta
D,
Bleakley
B,
Gupta
RK.
Dragon's
blood:
botany,
chemistry
and
therapeutic
uses.
J
Ethnopharmacol
2008;
115:
361−380
[4] Gibbs
A,
Green
C,
Doctor
VM.
Isolation
and
anticoagulant
properties
of
polysaccharides
of
Typha
Augustata
and
Daemonorops
species.
Thromb
Res
1983;
32:
97−108
[5] Tsai
WJ,
Hsieh
HT;
Chen
CC,
Chen
CF.
Studies
on
the
vasoactive-‐antithrombotic
effect
of
Draconis
Resina
and
its
components.
J
Chin
Med
1995;
6:
14
[6] Tsai
WJ,
Hsieh
HT,
Chen
CC,
Kuo
YC,
Chen
CF.
Characterization
of
the
antiplatelet
effects
of
(2S)-‐5-‐methoxy-‐6-‐methylflavan-‐7-‐ol
from
Draconis
Resina.
Eur
J
Pharmacol
1998;
346:
103−110
[7] Yi
T,
Chen
HB,
Zhao
ZZ,
Yu
ZL,
Jiang
ZH.
Comparison
of
the
chemical
profiles
and
anti-‐
platelet
aggregation
effects
of
two
“Dragon's
Blood”
drugs
used
in
traditional
Chinese
medicine.
J
Ethnopharmacol
2011;
133:
796−802
[8] Rao
GSR,
Gerhart
MA,
Lee
RT,
Mitscher
LA,
Drake
S.
Antimicrobial
Agents
From
Higher
Plants.
Dragon's
Blood
Resin.
J
Nat
Prod
1982;
45:
646−648
P489
Two
new
acylated
flavonol
glycosides
from
Mimosa
pigra
leaves
Chinedu
J.
Okonkwo1,
Tochukwu
J.
N.
Okonkwo2,6,
Ozadheoghene
E.
Afieroho3,
Blessing
O.
Okonkwo3,
Charles
O.
Esimone4,
Obioma
U.
Njoku5,
Peter
Proksch
1
Biochemistry
Department,
Faculty
of
Science,
University
of
Port
Harcourt,
500004
Port
Harcourt,
Nigeria,
2
Pharmaceutical
and
Medicinal
Chemistry
Department,
Faculty
of
Pharmaceutical
Sciences,
University
of
Port
Harcourt,
500004
Port
Harcourt,
Nigeria,
3
Pharmacognosy
and
Phytotherapy
Department,
Faculty
of
Pharmaceutical
Sciences,
University
of
Port
Harcourt,
500004
Port
Harcourt,
Nigeria,
4
Pharmaceutical
Microbiology
and
Biotechnology
Department,
Faculty
of
Pharmaceutical
Sciences,
Nnamdi
Azikiwe
University,
Awka,
Anambra
State,
Nigeria,
5
Biochemistry
Department,
Faculty
of
Biological
Sciences,
University
of
Nigeria,
Nsukka,
Enugu
State,
Nigeria,
6Institute
of
Pharmaceutical
Biology
and
Biotechnology,
Heinrich
Heine
University
Düsseldorf,
40225
Düsseldorf,
Germany
Mimosa
pigra
L.
(Fabaceae-‐Mimosoideae)
is
a
large
shrub
native
to
tropical
America
and
eco-‐
nomically
important
as
a
medicinal
plant;
it
is
also
an
invasive
plant
and
a
noxious
weed.
M.
pigra
leaves
were
air-‐dried,
pulverised
and
extracted
with
96
%
ethanol.
The
ethanol
extract
(MPEE)
was
partitioned
with
diethyl
ether
(DEE)
and
ethyl
acetate
(EA)
in
succession
to
yield
DEE
fraction
(DEEF),
EA
fraction
(EAF)
and
EA
insoluble
fraction
(EAIF).
Sephadex
LH-‐20
gel
filtration
chromatography
(GFC)
of
the
EAF
and
semi-‐preparative
HPLC
purification
of
the
GFC
sub-‐fractions
afforded
two
new
acylated
flavonol
glycosides
[myricetin
(2”-‐O-‐galloyl)-‐3-‐
O-‐α-‐L
rhamnopyranose,
I;
and
quercetin
(2”-‐O-‐galloyl)-‐3-‐O-‐α-‐L-‐rhamnopyranoside,
II]
[1]
in
M.
pigra.
Alongside
three
other
flavonol
glycosides
(myricetin
3-‐O-‐α-‐L-‐rhamnopyranoside,
III;
quercetin
3-‐O-‐α-‐L-‐rhamnopyranoside,
IV;
and
quercetin
3-‐O-‐α-‐L-‐arabinopyranoside,
V)
[2].
The
structures
of
compounds
I–V
were
assigned
by
ultraviolet/visible
(UV)
spectroscopy,
1D
and
2D
1H
and
13C
NMR
spectroscopy
and
liquid
chromatography-‐electrospray-‐mass
spec-‐
trometry
(LC-‐ESI-‐MS).
Our
findings
may
provide
important
insight
into
the
understanding
of
the
evolutionary
trend
of
the
medicinal
plant
[3],
its
medicinal
profile
and
ethnomedicinal
uses
[4]
as
well
as
the
mechanism
and
mode
of
its
confirmed
pharmacological
actions.
Acknowledgements:
Dr.
Tochukwu
J.
N.
Okonkwo
is
grateful
to
The
World
Academy
of
Science
(TWAS)
–
for
the
advancement
of
sciences
in
developing
countries
–
and
the
German
Research
Foundation
(DFG).
For
the
2014/2015
TWAS-‐DFG
Post-‐Doctoral
Visiting
Scientist
Research
Fellowship
Award
(Ref.
3240278143)
at
the
Institute
of
Pharmaceutical
Biology
and
Biotechnology,
Heinrich-‐Heine-‐University,
Düsseldorf,
Germany.
Keywords: Acylated flavonol glycosides, Mimosoideae subfamily, Mimosa pigra L.
References:
[1] Saldanha
L,
Vilegas
W,
Dokkedal
A.
Characterization
of
Flavonoids
and
Phenolic
Acids
in
Myrcia
bella
Chambers
Using
FIA-‐ESI-‐IT-‐MS
and
HPLC-‐PAD-‐ESI-‐IT-‐MS
Combined
with
NMR.
Molecules
2013;
18:
8402–8416
[2] Ablajan
K,
Abliz
Z,
Shang
X,
He
J,
Zhang
R,
Shi
J.
Structural
characterization
of
flavonol
3,7-‐
di-‐O-‐glycosides
and
determination
of
the
glycosylation
position
by
using
negative
ion
electrospray
ionization
tandem
mass
spectrometry.
J
Mass
Spectrom
2006;
41:
352–360
[3] Sulaiman
S.
Isolation
and
characterisation
of
flavonoids
from
Mimosa
pigra:
New
acylated
flavonols
from
Mimosa
pigra.
Project
Report.
USM.
http://eprints.usm.my/313/1/Isolation_And_Characterization_Of_Flavonoids_From_Mim
osa_Pigra.pdf
[4] Okonkwo
T,
Osadebe
P,
Proksch
P.
Bioactive
phenylpropanoids,
phenolic
acid
and
phy-‐
tosterol
from
Landolphia
owariensis
P.
Beauv
Stringy
Seed
Pulp.
Phytother
Res
2016;
30:
78–83.
P490
Comparison
of
genotoxicity
potentials
between
crude
Thai
bee
pollen
and
its
extracts
Treetip
Ratanavalachai1,
Sumon
Thitiorul2,
Wantha
Jenkhetkan3,
Chalerm
Jansom4,
Arunporn
Itharat5
1
Division
of
Biochemistry,
Department
of
Preclinical
Science,
Faculty
of
Medicine,
Thammasat
University,
12120,
Pathumthani,
Thailand,
2
Division
of
Anatomy,
Department
of
Preclinical
Science,
Faculty
of
Medicine,
Thammasat
University,
12120,
Pathumthani,
Thailand,
3
Ph.D.
graduate
program
in
Biochemistry
and
Molecular
Biology,
Faculty
of
Medicine,
Thammasat
University,
12120,
Pathumthani,
Thailand,
4
Research
center,
Faculty
of
Medicine,
Thammasat
University,
12120,
Pathumthani,
Thailand,
5
Department
of
Applied
Thai
Traditional
medicine,
Faculty
of
Medicine,
Thammasat
University,
12120,
Pathumthani,
Thailand
Bee
pollen
is
traditionally
used
as
a
food
supplement
and
a
natural
medicine [1].
However,
its
genotoxicity
and
cytotoxicity
studies
are limited.
This
study
investigated
the
genotoxicity
and
cytotoxicity
of
commercial
crude
bee
pollen
(CBP)
from
Northern
Thailand
using
in
vitro
sister
chromatid
exchange
(SCE)
assay
in
human
lymphocytes.
The
lipid
fractions
(LF)
and
defatted
fractions
(DF)
obtained
from
CBP-‐Soxhlet
extraction
using
diethyl
ether,
were also
analysed.
Human
lymphocytes
were
treated
with
CBP
at
5,
50,
500,
and
5000
ng/ml,
LF
at
1.25,
12.5,
125
and
1250
ng/ml
and
DF
at
5,
50,
500,
and
5000
ng/ml
for
three
hours
and
were
then
harvested,
stained
and
scored
for
SCE.
RPMI
culture
medium
and
doxorubicin
were
used
as
negative
and
positive
controls,
respectively.
Results
revealed
that
only
CBP
at
5000
ng/ml
was
cytotoxic
to
human
cells
as
few
mitotic
cells
were
detected.
SCE
levels
were
significantly
increased
in
human
lymphocytes
treated
with
5
and
500
ng/ml
CBP
compared
to
the
negative
control
(p<0.05),
indicating
that
the
genetic
damage
was
induced
in
these
CBP
groups.
Treatment
with
LF
at
1.25
ng/ml
significantly
increased
SCE
levels,
but
this
genotoxicity
was
not
observed
in
the
higher
concentrations.
Treatment
with
DF
at
all
concentrations
significantly
induced
genetic
damage.
Chemical
composition
analyses
revealed
that
flavonoid
and
phenolic
acid
contents
were
2.5-‐
and
8-‐fold
higher
in
DF
than
in
LF,
respectively.
Interestingly,
LF
contained
14
times
higher
amounts
of
ferulic
acid
than
DF
did.
Moreover,
vitamin
E
was
only
detected
in
LF.
In
summary,
DF
at
all
doses
induced
significant
genotoxicity
while
CBP
at
5
and
500
ng/ml
as
well
as
LF
only
at
1.25
ng/ml
induced
signifi-‐
cant
genetic
damage.
The
underlying
mechanisms
may
involve
their
different
composition
ratio
such
as
ferulic
acid,
and
vitamin
E.
Further
studies
of
these
molecular
mechanisms
underlying
genotoxicity
should
be
encouraged.
Acknowledgements:
This
work
is
supported
by
Research
Fund,
Thammasat
University
2015
and
Research
Fund,
National
Research
University
Project
of
Thailand.
Office
of
Higher
Education
Commission
and
Thammasat
University,
Thailand.
Keywords:
Bee
pollen,
sister
chromatid
exchange,
genotoxicity,
ferulic
acid,
human
lympho-‐
cyte
References:
[1] Komosinska-‐Vassev
K,
Olczyk
P,
Kazmierczak
J,
Mencner
L,
Olczyk
K.
Bee
pollen:
chemical
composition
and
therapeutic
application.
Evid
Based
Complement
Alternat
Med
2015;
297425
P491
Antimutagenic
activity
of
n-‐tetracosanol,
eicosanoic
acid
and
ar-‐
junolic
acid
isolated
from
Combretum
microphyllum
(Combreta-‐
ceae)
Tshepiso
J.
Makhafola1,2,
Esameldin
E.
Elgorashi
3,4,
Lyndy
J.
McGaw1,
Maurice
D.
Awouafack1,
Luc
Verschaeve5,6,
Jacobus
N.
Eloff1
1
Department
of
Paraclinical
Sciences,
Phytomedicine
Programme,
University
of
Pretoria,
University
of
Pretoria,
Onderstepoort,
0110,
South
Africa;
2
Department
of
Life
and
Consumer
Sciences,
University
of
South
Africa,
Florida,
1710,South
Africa;
3
Toxicology
and
Ethnoveterinary
Medicine,
Food,
Feed
and
Vet-‐
erinary
Public
Health,
ARC-‐Onderstepoort
Veterinary
Institute,
Onderstepoort
0110,
South
Africa;
4
De-‐
partment
of
Paraclinical
Sciences,
Faculty
of
Veterinary
Science,
,
Private
Bag
X04,
Onderstepoort
0110,
South
Africa,
5
Scientific
Institute
of
Public
Health,
Rue
Juliette
Wytsmanstreet
14,1050
Brussels,
Belgium,
6
Department
of
Biomedical
Sciences,
University
of
Antwerp,
Universiteitsplein
1,
B-‐2610
Wilrijk,
Belgium
The
possibility
of
moderating
the
response
of
cells
to
a
particular
mutagen
by
phytomedicines
opens
new
horizons
in
cancer
control.
On
this
basis,
the
research
for
antimutagens
presents
many
possibilities
for
the
discovery
of
new
anticarcinogenic
substances
[1,
2].
The
methanol
extract
of
C.
microphyllum
was
evaluated
for
antimutagenicity
in
the
Ames/microsome
assay
using
Salmonella
typhimurium
TA98,
TA100
and
TA102
without
metabolic
activation.
A
bioassay-‐guided
fractionation
of
the
crude
extracts
led
to
the
isolation
of
three
compounds:
n-‐
tetracosanol,
eicosanoic
acid
and
arjunolic
acid.
Arjunolic
acid
was
the
most
active
in
all
three
tested
strains
with
percentage
antimutagenicity
of
up
to
41.92
±
9.59%,
35.84
±
1.45%
and
43.78
±
0.18%
in
S.
typhimurium
TA98,
TA100
and
TA102
respectively
at
the
highest
concentration
tested,
followed
by
eicosanoic
acid
and
lastly
n-‐tetracosanol.
The
compounds
were
assessed
for
antioxidant
activity
using
the
quantitative
2,2-‐diphenyl-‐1-‐picrylhydrazyl
(DPPH)-‐free
radical
scavenging
method.
Arjunolic
acid
was
the
only
compound
with
pronounced
antioxidant
activity
(measured
as
DPPH-‐free
scavenging
activity)
with
EC50
value
of
0.51
µg/ml.
Furthermore,
the
isolated
compounds
were
assessed
for
their
potential
toxic
effects
in
the
MTT
assay
using
human
hepatocytes.
All
three
compounds
were
tested
at
concentrations
ranging
from
200µg/ml
to
10µg/ml;
with
20µg/ml
being
the
highest
concentration
tested.
The
compounds
were
not
toxic
with
EC50
values
>200
µg/ml
for
n-‐
tetracosanol
and
eicosanoic
acid
and
106.39µg/ml
for
arjunolic
acid.
Based
on
findings
from
this
study,
compounds
from
C.
microphyllum
protect
against
4-‐NQO
and
MMC
induced
mutations
as
evident
in
the
Ames
test.
Taking
into
account
that
genotoxicity
involving
gene
mutations
plays
a
major
role
in
cancer
initiation
[3].
C.
microphyllum
has
potential
in
cancer
prevention
as
it
inhibits
these
genotoxic
end-‐points.
The
antimutagenic
activity
of
arjunolic
acid,
at
least
in
part,
may
be
attributed
to
its
antioxidant
activity
which
results
in
the
detoxification
of
reactive
oxygen
species
produced
during
mutagenesis
[4].
Acknowledgements:
National
Research
Foundation,
University
of
Pretoria
and
University
of
South
Africa
for
financial
support.
Curators
of
Pretoria
National
Botanical
Gardens
allowed
us
to
collect
plant
materi-‐
al.
Keywords: Combretum microphyllum, n-‐tetracosanol, eicosanoic acid, arjunolic acid, antimu-‐
tagenicity, cytotoxicity, antioxidant activity
References:
[1] De
Flora
S,
Izzotti
A,
D’Agostini
F,
Balansky
R.
Mechanisms
of
n-‐acetylcysteine
in
the
pre-‐
vention
of
DNA
damage
and
cancer,
with
specific
reference
to
smoking-‐related
end
points.
Carcinogenesis
2001;
22:
999−1013
[2] Ferguson
LR,
Zhu
S,
Harris
PJ.
Antioxidant
and
antigenotoxic
effects
of
plant
cell
wall
hy-‐
droxycinnamic
acids
in
cultured
HT-‐29
cells.
Mol
Nutr
Food
Res
2005;
49:
585−693
[3] Ames
BN.
Dietary
carcinogens
and
anticarcinogens:
oxygen
radicals
and
degenerative
dis-‐
eases.
Science
1983;
221:
1256−1263
[4] Hemalatha
T,
Pulavendran
S,
Balachandran
C,
Manohar
BM,
Puvanakrishnan
R.
Arjunolic
acid:
A
novel
phytomedicine
with
multifunctional
therapeutic
applications.
Indian
J
Exp
Biol
2010;
48:
238−247
P492
Evaluation
for
the
anti-‐leukemic
activities
and
28-‐day
subacute
toxicity
of
ethanolic
extracts
from
the
fruiting
bodies
of
dish-‐
cultured
Antrodia
cinnamomea
Tung-‐Ying
Wu1,
Mohamed
El-‐Shazly2,
Ying-‐Chi
Du3,
Yu-‐Ming
Hsu3,
Kuei-‐Hung
Lai3,
Mei-‐Chin
Lu4,
5,
Yang-‐Chang
Wu1,
6
1
Chinese
Medicine
Research
and
Development
Center,
China
Medical
University
Hospital,
Taichung
404,
Taiwan;
2
Department
of
Pharmacognosy
and
Natural
Products
Chemistry,
Faculty
of
Pharmacy,
Ain-‐
Shams
University,
Cairo,
Egypt;
3
Graduate
Institute
of
Natural
Products,
College
of
Pharmacy,
Kaohsiung
Medical
University,
Kaohsiung
807,
Taiwan;
4
Graduate
Institute
of
Marine
Biology,
National
Dong
Hwa
University,
Pingtung
944,
Taiwan;
5
National
Museum
of
Marine
Biology
&
Aquarium,
Pingtung
944,
Tai-‐
wan;
6School
of
Pharmacy,
College
of
Pharmacy,
China
Medical
University,
Taichung
404,
Taiwan
Antrodia
cinnamomea
Chang
&
Chou
(AC)
[Fomitopsidaceae],
an
endemic
species,
has
long
been
used
as
a
chemopreventive
mushroom
in
traditional
Taiwanese
medicine.
Among
differ-‐
ent
AC
components,
triterpenoids
are
considered
the
most
therapeutically
attractive
constitu-‐
ents
due
their
cytotoxic
and
anti-‐inflammatory
activities
[1].
In
this
study,
we
report
a
stand-‐
ardized
triterpenoids-‐dependent
analysis
platform
for
the
five
available
AC
marketed
prod-‐
ucts
by
using
HPLC-‐PDA.
Among
them,
the
ethanolic
extracts
from
fruiting
bodies
of
dish-‐
cultured
AC
(DC-‐EEAC)
had
a
similar
triterpenoids
profile
compared
to
that
of
wild
AC
(EEAC).
During
the
evaluation
of
anti-‐leukemic
activity
of
DC-‐EEAC
against
leukemia
cancer
cell
line
(Molt
4),
the
results
revealed
that
the
activation
of
DNA
damage
and
apoptosis
biomarkers
by
DC-‐EEAC.
In
the
xenograft
animal
model,
DC-‐EEAC
resulted
in
a
marked
decrease
of
tumour
weight
and
size
without
any
significant
decrease
in
mice
body
weights.
Furthermore,
test
on
the
28-‐day
oral
toxicity
of
DC-‐EEAC
in
male
and
female
nude
mice
exhibited
no
significant
changes
in
histopathological
examination
and
no
abnormal
changes
were
observed
in
body
weight
and
serum
biochemistry
parameters
compared
to
the
control
group.
Our
results
showed
that
fruiting bodies from the dish-cultured AC
has
a
significant
anti-‐proliferative
activi-‐
ty
and
primary
safety
profile
could
be
considered
as
one
of
alternative
AC
sources.
Acknowledgements:
This
work
was
supported
by
grants
from
ministry
of
science
and
technology
of
Tai-‐
wan
awarded
to
Mei-‐Chin
Lu
and
Yang-‐Chang
Wu.
Keywords: Antrodia cinnamomea, triterpenoids, dish-‐cultured, anti-‐proliferative activity,
subacute toxicity
References:
[1] Lu
MC,
El-‐Shazly
M,
Wu
TY,
Du
YC,
Chang
TT,
Chen
CF,
Hsu
YM,
Lai
KH,
Chiu
CP,
Chang
FR,
Wu
YC.
Recent
research
and
development
of
Antrodia
cinnamomea.
Pharmacol
Ther
2013;
139:
124−156
P493
Bioactive
constituents
from
the
termite
nest-‐derived
medicinal
fungus
Xylaria
nigripes
Jung-‐Chun
Chang1,
George
Hsiao2,
Ruo-‐Kai
Lin1,
Yueh-‐Hsiung
Kuo3,
Yu-‐Min
Ju4,
Tzong-‐Huei
Lee5
1
Graduate
Institute
of
Pharmacognosy,
Taipei
Medical
University,
250
Wuxing
Street,
Taipei
110,
Taiwan,
2
Graduate
Institute
of
Medical
Science
and
Department
of
Pharmacology,
College
of
Medicine,
Taipei
Medical
University,
250
Wuxing
Street,
Taipei
110,
Taiwan,
3
Department
of
Chinese
Pharmaceutical
Sci-‐
ences
and
Chinese
Medicine
Resources,
China
Medical
University,
91
Hsueh-‐Shih
Road,
Taichung
404,
Tai-‐
wan,
4
Institute
of
Plant
and
Microbial
Biology,
Academia
Sinica,
128
Academia
Road,
Section
2,
Nankang,
Taipei
115,
Taiwan,
5
Institute
of
Fisheries
Science,
National
Taiwan
University,
1
Roosevelt
Road,
Section
4,
Taipei
106,
Taiwan
Six
new
eremophilane-‐type
sesquiterpenes,
namely
nigriterpenes
A−F
(1−6),
and
one
new
phenolic
compound,
namely
2-‐hyroxymethyl-‐3-‐pentylphenol
(7),
along
with
fomannoxin
al-‐
cohol,
3-‐butyl-‐7-‐hydroxyphthalide,
scytalone,
and
fomannoxin
were
isolated
from
the
ethyl
acetate
extracts
of
the
fermented
broths
of
termite
nest-‐derived
Xylaria
nigripes,
which
has
long
been
used
as
the
traditional
Chinese
medicine
for
treating
insomnia
and
depression
[1,
2].
The
structures
of
all
the
compounds
were
elucidated
based
on
spectroscopic
data
analysis
and
compared
with
literatures.
The
absolute
configurations
of
1−6
were
established
by
NOESY,
and
compared
with
the
literatures.
All
the
pure
compounds
were
evaluated
their
effects
on
lipopolysaccharide
(LPS)-‐induced
inducible
nitric
oxide
synthase
(iNOS),
cyclooxygenase-‐2
(COX-‐2)
expression,
and
NO
production
on
murine
brain
microglial
BV-‐2
cells
[3].
Of
the
com-‐
pounds
tested,
both
nigriterpene
C
(3)
and
fomannoxin
alcohol
exerted
significant
inhibition
on
these
responses
without
any
significant
cellular
toxicity.
The
most
potent
compound
3
ex-‐
hibited
concentration-‐dependent
inhibition
on
NO
production,
iNOS
and
COX-‐2
expression
with
IC50
values
of
21.7±
4.9,
5.6
±
0.7,
and
12.4
±
3.9
µM,
respectively.
These
results
indicated
that
the
potential
anti-‐inflammatory
effects
of
nigriterpene
C
(3)
and
fomannoxin
alcohol
on
murine
brain
microglial
BV-‐2
cells
may
provide
a
rationale
for
the
traditional
medical
uses
of
X.
nigripes
on
chronic
brain
inflammation-‐involved
diseases.
Acknowledgments:
This
work
was
supported
by
grants
from
the
Ministry
of
Science
and
Technology
of
the
Republic
of
China.
Keywords: Xylaria nigripes, termite nest, sesquiterpene, antiinflammation, BV-‐2
References:
[1] Lin
Y,
Wang
XY,
Ye
R,
Hu
WH,
Sun
SC,
Jiao
HJ,
Song
XH,
Yuan
ZZ,
Zheng
YY,
Zheng
GQ,
He
JC.
Efficacy
and
safety
of
Wuling
capsule,
a
single
herbal
formula,
in
Chinese
subjects
with
in-‐
somnia:
a
multicenter,
randomized,
double-‐blind,
placebo-‐controlled
trial.
J
Ethnophar-‐
macol
2013;
145:
320-327
[2] Li
D,
Zheng
J,
Wang
M,
Feng
L,
Liu
Y,
Yang
N,
Zuo
P.
Wuling
powder
prevents
the
depres-‐
sion-‐like
behavior
in
learned
helplessness
mice
model
through
improving
the
TSPO
me-‐
diated-‐mitophagy.
J
Ethnopharmacol
2016;
186:
181-‐188
[3] Chou
YC,
Sheu
JR,
Chung
CL,
Chen
CY,
Lin
FL,
Hsu
MJ,
Kuo
YH,
Hsiao
G.
Nuclear-‐targeted
in-‐
hibition
of
NF-‐kappaB
on
MMP-‐9
production
by
N-‐2-‐(4-‐bromophenyl)ethyl
caffeamide
in
human
monocytic
cells.
Chem
Biol
Interact
2010;
184:
403-412
P494
Phenolic
compounds
of
Ornithogalum
pyrenaicum
together
with
antimicrobial
and
antioxidant
studies
Nuriye
Korkmaz
1,
Sıla
Özlem
Şener
2,
Merve
Badem1,
Ufuk
Özgen1,
Rezzan
Aliyazicioglu2,
Şen-‐
gül
Alpay
Karaoğlu3
1
Department
of
Biochemistry,
Karadeniz
Technical
University,
Faculty
of
Pharmacy,
61080
Trabzon,
TURKEY,
2
Department
of
Pharmacognosy,
Karadeniz
Technical
University,
Faculty
of
Pharmacy,
61080
Trabzon,
TURKEY,
3
Department
of
Biology,
Recep
Tayyip
Erdoğan
University,
Faculty
of
Science,
53100
Rize,
TURKEY
Ornithogalum
genus
(Hyacinthaceae)
was
represented
by
160
species
[1]
and
distributed
in
Europe,
Asia,
Africa
and
Madagascar
[2].
It
has
23
species
in
Turkey
[3].
By
means
of
this
study,
qualitative
and
quantitative
analysis
of
phenolic
compounds
including
13
phenolic
standarts
by
RP-‐HPLC
and
antioxidant
capacity
by
three
methods
including
DPPH,
FRAP
and
total
phenolic
contents
were
examined
on
O.
pyrenaicum.
In
addition,
antimicrobial
activity
of
the
plant
was
researched
on
9
microorganism
with
agar
diffusion
method.
As
a
result
of
the
study,
p-‐coumaric
acid
and
benzoic
acid
among
phenolic
compounds
were
detected
and
me-‐
dium
antioxidant
capacity
was
observed.
The
IC50
value
for
DPPH
assay
has
been
found
as
2,2186±0,0418
(mg/mL),
FRAP
value
is
129±3,7859
(μM
Trolox/g
sample),
and
total
phenolic
content
value
is
5,7
±
0,2645
mg
gallic
acid
per
gram
sample
for
the
aerial
parts
of
the
plant.
Via
antimicrobial
activity
study,
it
was
determined
that
O.
pyrenaicum
had
considerable
activi-‐
ty
on
Yersinia
pseudotuberculosis,
Staphylococcus
aureus
and
moderate
activity
on
Pseudomo-‐
nas
aeruginosa
and
Mycobacterium
smegmatis.
The
study
may
be
guide
for
further
phyto-‐
chemical
studies
on
the
plant.
Keywords:
Ornithogalum
pyrenaicum,
RP-‐HPLC,
antioxidant
capacity,
antimicrobial
activity
References:
[1] Manning
JC,
Forest
F,
Devey
DS,
Fay
MF,
Goldblatt
P.
A
molecular
phylogeny
and
a
revised
classification
of
Ornithogaloidea
(Hyacinthaceae)
based
on
an
analysis
of
four
plastid
DNA
regions.
Taxon
2009;
58:
77-‐107
[2] Mutlu
B,
Karakuş
S.
A
new
species
of
Ornithogalum
(Hyacinthaceae)
from
East
Anatolia,
Turkey.
Turk
J
Bot
2012;
36:
125-‐133
[3] Cullen
J.
Parentucellia
Viv.
in
Flora
of
Turkey
and
the
East
Aegean
Islands,
8th
edition.
Ed-‐
inburg:
Edinburg
University
Press:
1984:
227-‐227
P495
Bioactivity
studies
of
algal
extracts
from
Turkey’s
Mediterranean
coast
Yasin
Genc1,
Secil
Sarikaya
Aydin1,
Melek
Sertdemir2,
Ozge
Demir2,
Emine
S.
Okudan3,
Selin
Salin3,
Belma
Konuklugil2,
Ummuhan
Sebnem
Harput1
1Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Hacettepe
University,
06100,
Sihhiye,
An-‐
kara,
Turkey,
2Biotechnology
Institute,
Ankara
University,
06100,
Tandoğan,
Ankara,
Turkey,
3Faculty
of
Fisheries,
Mustafa
Kemal
University,
31030,
Hatay.
Turkey
is
a
country
of
peninsula
with
nearly
8300
km
marine
coasts
and
the
rich
biodiversity
of
this
region,
however,
has
not
been
investigated
completely
from
the
view
point
of
biological
active
extracts
[1].
In
this
study,
eight
different
marine
algae
distributed
in
Hatay,
Mediterra-‐
nean
Coasts,
were
investigated
for
their
superoxide
(SO)
radical
scavenging
effects
and
cyto-‐
toxic
activities
against
Hep-‐2
(Human
Epidermoid
Carcinoma)
cancer
cell
line.
SO
radical
scavenging
activity
was
tested
in
the
concentration
range
of
100-‐800
µg/mL.
While
Liagora
viscida
and
Padina
sp.
(IC50
>
800
µg/mL)
extracts
showed
dose
dependent
SO
radical
scav-‐
enging
activity
but
lower
than
that
of
ascorbic
acid
and
quercetin,
other
tested
extracts
did
not
show
any
activity
in
tested
concentrations.
On
the
other
hand,
cytotoxic
activity
of
the
ex-‐
tracts
was
tested
by
MTT
method
and
methanoic
extracts
of
Padina
sp.,
Liagora
viscida
and
Laurencia
obtusa
(IC50
˂
10
µg/mL)
showed
dose
dependent
and
significant
cytotoxic
activity.
However,
methanolic
extracts
of
Dictyota
dichotoma,
Hypnea
musciformis,
Codium
fragile,
Pali-‐
sada
perforata
and
Gracilaria
bursa-‐pastoris
(IC50:
10-‐100
µg/mL)
showed
moderate
cytotoxi-‐
city.
Based
on
the
bioactivities
of
the
methanolic
extract
of
Padina
sp.
and
Liagora
viscida,
phytochemical
studies
will
be
initiated
to
find
the
active
components
in
the
extracts
of
these
species.
Acknowledgements:
Ummuhan
Sebnem
Harput
has
been
supported
TUBA-‐GEBİP/2013
award.
Keywords:
Algae,
superoxide,
cytotoxicity.
References:
[1] Süzgeç-‐Selçuk
S,
Meriçli
AH,
Güven
KC.
Kaiser
M,
Casey
R,
Hingley-‐Wilson
S,
Lalvani
A,
Tasdemir
D.
Evaluation
of
Turkish
seaweeds
for
antiprotozoal,
antimycobacterial
and
cy-‐
totoxic
activities.
Phytother
Res
2011;
25:
778–783
P496
Bioassay
guided
isolation
studies
on
Incarvillea
emodi
(Royle
ex
Lindl.)
Chatterjee
Yasir
Ihtesham1,2,
Zeynep
Dogan1,
Vahap
Murat
Kutluay1,
Uzma
Khan2,
Iclal
Saracoglu1
1Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Hacettepe
University,
06100,
Sihhiye,
Ankara,
Turkey,
2Department
of
Botany,
Garden
Campus,
Hazara
University,
Mansehra,
Pakistan
Incarvillea
Juss.
is
notable
for
being
a
temperate
and
herbaceous
genus
of
the
primarily
tropi-‐
cal
and
woody
family
Bignoniaceae.
It
is
composed
of
16
species
worldwide
and
represented
in
Pakistan
by
one
native
species
[1,
2].
Aqueous
extracts
of
the
aerial
parts
(ABD-‐Ap)
and
the
roots
of
Incarvillea
emodi
collected
from
Abbottabad
(ABD-‐Rt)
and
the
aerial
parts
of
I.
emodi
collected
from
Kashmir
(K-‐Ap)
were
prepared,
separately.
Each
extract
was
tested
for
its
rad-‐
ical
scavenging
and
cytotoxic
activities
using
DPPH
radical
and
HEp-‐2
cell
line
respectively.
IC50
values
for
aqueous
extracts
for
DPPH
radical
were
found
to
be;
63.5
µg/mL,
50.7
µg/mL,
142.4
µg/mL,
respectively.
Cytotoxic
activity
of
ABD-‐Ap
and
K-‐Ap
was
tested
against
HEp-‐2
cell
lines
by
MTT
method
[3]
and
their
IC50
values
were
determined
as
109.4
and
211.6
µg/mL
respectively.
For
phytochemical
studies,
aqueous
extract
of
aerial
parts
(ABD-‐Ap)
of
I.
emodi
was
selected
on
the
basis
of
bioactivity
guided
fractionation
and
applied
to
polyamide
column
chromatography
for
fractionation.
Due
to
examination
of
radical
scavenging
and
cytotoxic
activities,
active
fractions
were
detected
and
isolation
studies
were
started
from
active
frac-‐
tions.
Serial
chromatographic
studies
resulted
in
the
isolation
of
five
compounds.
Four
phe-‐
nylethanoid
glycosides,
acteoside
(1),
2-‐O-‐acetylacteoside
(2),
martynoside
(3),
and
tubu-‐
loside
E
(4)
while
one
iridoid
glucoside,
plantarenaloside
(5)
were
identified
by
extensive
NMR
techniques.
Isolated
compounds
also
showed
high
radical
scavenging
activity
against
DPPH
radical.
O
CHO
OH
OH
R1 O
O O OH
O
OR3
O
HO OH
H3C O
HO OR2 H3C
O
O
HO
HO
OH
OH R
1
R2
R3
OH
-‐OH -‐H -‐H (1)
-‐OH -‐H -‐COCH3 (2)
-‐OCH3 -‐CH3 -‐H (3) (5)
-‐H -‐H -‐COCH3 (4)
Acknowledgements:
This
study
was
supported
by
The
Scientific
and
Technological
Research
Council
of
Turkey
(TUBITAK-‐
2216)
and
Higher
Education
Commission
of
Pakistan
(HEC).
The
authors
are
grateful
to
Prof.
Dr.
Toshiaki
Makino
(Nagoya
City
University,
Faculty
of
Pharmaceutical
Sciences,
Nagoya,
Japan)
for
recording
NMR
spectra.
Keywords: Incarvillea emodi, DPPH, cytotoxic activity, MTT method, NMR.
References:
[1] Jian-‐Jun
F,
Hui-‐Zi
J,
Yun-‐Heng
S,
Jiang-‐Jiang
Q,
Yan
W,
Ying
H,
Qi
Z,
Wei-‐Dong
Z.
Chemical
constituents
of
plants
from
the
genus
Incarvillea.
Chem
Biodivers
2009;
6:
818−826
[2] Flora
of
Pakistan.
http://www.tropicos.org/Project/Pakistan
[3] Saracoglu
I,
Inoue
M,
Calis
I,
Ogihara
Y.
Studies
on
constituents
with
cytotoxic
and
cyto-‐
static
activity
of
two
Turkish
medicinal
plants
Phlomis
armeniaca
and
Scutellaria
salviifo-‐
lia.
Biol
Pharm
Bull
1995;
18:
1396−1400
P497
Cytotoxicity
comparison
on
different
parts
of
three
Digitalis
spe-‐
cies
and
isolation
of
potential
bioactive
compounds
Vahap
Murat
Kutluay1,
Makoto
Inoue2,
U.
Sebnem
Harput1,
Iclal
Saracoglu1
1Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Hacettepe
University,
06100,
Ankara,
Turkey,
2Laboratory
of
Medicinal
Resources,
School
of
Pharmacy,
Aichi
Gakuin
University,
1-‐100
Kusumoto-‐cho,
Chikusa-‐ku Nagoya, Japan
In the Flora of Turkey, the genus Digitalis is represented by nine species [1]. Digitalis species
contain biologically active compounds such as cardenolides, phenylethanoid glycosides, fla-‐
vonoids, and anthraquinones. In this research three Digitalis species [Plantaginaceae]; D. da-‐
visiana Heywood, D. grandiflora Miller and D. viridiflora Lindley were studied in the point of
cytotoxicity. To determine the bioactive part of the plant, root and aerial parts were extracted
with methanol separately. Cytotoxic activity was performed by MTT method through 2 cancer
cell lines HEp-‐2 (Human larynx epidermoid carcinoma), HepG2 (Human hepatocelular carci-‐
noma) and a non-‐cancerous cell line 3Y1(rat fibroblast cell) [2]. Methanol extracts of the aeri-‐
al parts and the roots of three plants were used for cytotoxic activity tests. It was found that
these three plants have different potentials for cytotoxicity. Aerial parts were found to be
more active than the roots on tested cancer cell lines. In addition to this, D. davisiana aerial
parts extract (DD-‐APE) was found to be the most active among all. IC50 values of DD-‐APE were
50.8, 211.4 and 94.7 µg/mL respectively for HEp-‐2, HepG2 and 3Y1 cell lines. The tested ex-‐
tracts are more cytotoxic on HEp-‐2 cell line than HepG2 and 3Y1 cell lines. This selectivity is
important for the discovery of new anticancer agents. After evaluating the bioactivity test re-‐
sults, isolation studies started from active, phenylethanoid glycosides rich fractions. Repeated
column chromatography studies resulted in the isolation of five phenylethanoid glycosides.
Deacyllugrandoside (1), lugrandoside (2), isolugrandoside (3), maxoside (4) and 3,4-‐
dihydroxy-‐β-‐phenylethoxy-‐O-‐[β-‐glucopyranosyl-‐(1→3)]-‐O-‐[α-‐rhamnopyranosyl-‐(1→6)]-‐4-‐O-‐
(E)-‐feruloyl-‐β-‐glucopyranoside (5) were identified by extensive NMR techniques. Isolation
and cytotoxic activity studies on isolated compounds are still in progress.
OR3
O
R2O R O O OH
1
OH
OH
1 R1: H, R2:H, R3: β-‐Glc
2 R1: H, R2: (E)-‐Caffeoyl, R 3: β-‐Glc
3 R1: (E)-‐Caffeoyl, R 2:H, R3: β-‐Glc
Acknowledgements: Vahap Murat Kutluay was supported by research grant from The Scientific and
Technological Research Council of Turkey (2214/A, 2211). Authors are thankful to Prof. Dr. A. Nagatsu,
College of Pharmacy, Kinjo Gakuin University, Nagoya, Japan, for providing and recording NMR experi-‐
ments
Keywords: Digitalis, cytotoxicity, phenylethanoid glycosides
References:
[1]
Davis
PH.
Flora
of
Turkey
and
the
East
Eagean
Islands,
University
Press,
Edinburg,
1978,
Vol
6.
[2]
Saracoglu
I,
Inoue
M,
Calis
I,
Ogihara
Y.
Studies
on
constituents
with
cytotoxic
and
cyto-‐
static
activity
of
two
Turkish
medicinal
plants
Phlomis
armeniaca
and
Scutellaria
salviifo-‐
lia.
Biol
Pharm
Bull
1995;
18:
1396−1400
P498
Quantitative
analysis
of
polyphenols
and
antioxidant
activity
of
Croatian
populations
of
Erodium
cicutarium
(L.)
L'Herit
(Gerania-‐
ceae)
Vanja
Ljoljić
Bilić1,
Jadranka
Vuković
Rodríguez1,
Renata
Jurišić
Grubešić1,
Dario
Kremer2,
Živka
Juričić1
1
Department
of
Pharmaceutical
Analysis,
Faculty
of
Pharmacy
and
Biochemistry,
University
of
Zagreb,
Ante
Kovačića
1,
HR-‐10000
Zagreb,
Croatia,
2
Department
of
Pharmaceutical
Botany
with
Fran
Kušan
botanical
garden,
Faculty
of
Pharmacy
and
Biochemistry,
University
of
Zagreb,
Schrottova
37,
HR-‐10000
Zagreb,
Croatia
The
aim
of
this
study
was
screening
of
phytochemical
and
antioxidant
properties
of
aerial
parts
of
Erodium
cicutarium
(L.)
L'Herit.,
a
native
traditional
medicinal
plant,
from
four
locali-‐
ties
of
northern
(Buzin,
Trešnjevka),
central
(Plitvice)
and
eastern
part
of
Croatia
(Podvinje).
The
content
of
total
polyphenols
and
tannins
(TP
and
T;
Folin–Ciocalteu
assay)
[1],
total
fla-‐
vonoids
(TF;
colorimetric
assay
with
AlCl3)
[2]
and
total
hydroxycinnamic
derivatives
(THD;
colorimetric
assay
with
nitrite-‐molybdate
reagent)
[3]
in
plant
extracts
(aqueous,
ethanolic
and
methanolic)
was
quantified.
Well-‐established
assays
were
used
to
determine
the
antioxi-‐
dant
potency
of
the
extracts:
the
Ferric
Reducing/Antioxidant
Power
assay
(FRAP)
[4],
2,2-‐
diphenyl-‐1-‐picrylhydrazyl
(DPPH)
assay
[5],
and
2,2'-‐azinobis
(3-‐ethylbenzthiazoline-‐6-‐
sulphonic
acid)
(ABTS)
assay
[6].
The
contents
of
TP,
T,
TF,
and
THD
in
aerial
parts
of
the
in-‐
vestigated
plant
populations
significantly
varied
depending
on
the
collection
site,
and
were
in
the
range
of
4.78%
-‐
12.85%
(TP),
3.23
-‐
5.80%
(T),
0.42
-‐
1.09%
(TF),
and
1.08
-‐
2.59%
(THD).
The
sample
collected
from
locality
Plitvice
exhibited
significantly
higher
content
of
TP
(12.85±0.33%),
T
(5.80%±0.34%),
TF
(1.09%±0.04%),
and
THD
(2.59±0.12%)
in
comparison
with
other
examined
samples.
The
analyzed
extracts
exhibited
high
antioxidant
capacity
ac-‐
cording
to
all
three
assays
with
the
highest
for
methanol
extracts.
The
highest
results
of
anti-‐
oxidant
assays
were
obtained
for
Plitvice
population
of
E.
cicutarium
(L.)
L'Herit.,
i.e.
28.15
µmol
Fe2+/g
DW*
(FRAP),
1.94
mmol/L
TEAC**
(DPPH),
and
1.63
mM
TEAC
(ABTS).
These
results
contribute
to
the
scientific
study
of
phytotherapeutic
potential
of
the
investigated
plant
species,
especially
in
relation
to
bioactive
polyphenolic
substances,
and
give
an
impetus
for
further
in
vitro
and
in
vivo
biological
studies.
Note:
*DW
=
dry
weight;
**TEAC
=
Trolox
equivalent
antioxidant
capacity
Keywords:
Erodium
cicutarium
(L.)
L'Herit,
antioxidant
capacity,
polyphenols
References:
[1] Jurišić
Grubešić
R,
Vuković
J,
Kremer
D,
Vladimir-‐Knežević
S.
Spectrophotometric
method
for
polyphenols
analysis:
prevalidation
and
application
on
Plantago
L.
species.
J
Pharm
Biomed
Anal
2005;
39:
837-‐842
[2] Jurišić
Grubešić
R,
Vuković
J,
Kremer
D,
Vladimir-‐Knežević
S.
Flavonoid
content
assay:
Prevalidation
and
application
on
Plantago
L.
species.
Acta
Chim
Slov
2007;
54:
397-‐406
[3] European
Pharmacopoeia,
6th
Edition,
Vol.
2,
Strasbourg:
Councile
of
Europe,
2007
[4] Benzie
IFF,
Strain
JJ.
Ferric
reducing/antioxidant
power
assay:
direct
measure
of
total
an-‐
tioxidant
activity
of
biological
fluids
and
modified
version
for
simultaneous
measurement
of
total
antioxidant
power
and
ascorbic
acid
concentration.
Met
Enzymol
1999;
299:
15–
27
[5] Brand-‐Williams
W,
Cuvelier
ME,
Berset
C.
Use
of
a
free
radical
method
to
evaluate
antioxi-‐
dant
activity.
LWT–Food
Sci
Technol
1995;
28:
25–30
[6] Re
R,
Pellegrini
N,
Proteggente
A,
Pannala
A,
Yang
M,
Rice-‐Evans
C.
Antioxidant
activity
applying
an
improved
ABTS
radical
cation
decolorization
assay.
Free
Radic
Biol
Med
1999;
26:
1231–1237
P499
Secondary
metabolites
in
honey
and
their
inhibitory
activity
on
matrix
metalloproteinases
Boznou
Vasiliki,
Polychronopoulos
Panagiotis,
Michalis
Zervos,
Stavropoulou
Maria-‐Ioanna,
Aligiannis
Nektarios
Department
of
Pharmacognosy
and
Natural
Product
Chemistry,
Faculty
of
Pharmacy,
University
of
Ath-‐
ens,
Panepistimiopolis,
Zografou,
GR-‐15771,
Athens,
Greece
Honey
is
a
viscous,
supersaturated
sugar
solution
derived
from
nectar,
and
gathered
and
modified
by
the
honeybee,
Apis
mellifera
[1].
It
has
been
used
as
a
traditional
medicine
for
centuries
by
different
cultures.
Honey
possesses
antimicrobial
properties
against
a
broad
range
of
microbes,
and
promotes
wound
healing
[2].
In
the
present
study,
after
a
first
elimina-‐
tion
of
sugars,
the
chemical
profile
of
10
Greek
monofloral
honeys
(e.g.
thyme,
chestnut,
fir
honeydew,
pine,
orange,
heather,
strawberry
tree)
was
investigated
by
HPTLC,
which
permits
the
automatic,
rapid
and
low
cost
evaluation
of
complex
herbal
mixtures.
Furthermore,
their
radical
scavenging
activity
was
tested
with
the
aid
of
a
DPPH
assay.
Two
honey
samples,
a
strawberry
tree
and
a
heather
honey,
were
further
processed,
and
major
compounds
were
purified
by
MPLC.
Unedone
was
obtained
from
strawberry
tree
honey
and
the
flavonoids
quercitrin
and
naringenin
from
heather
honey,
while
two
isomers
of
abscisic
acid
(cis,
trans-‐
ABA
and
trans,
trans-‐ABA)
were
isolated
from
both
samples.
Subsequently,
raw
honeys
and
pure
compounds
were
evaluated
for
their
ability
to
inhibit
the
matrix
metalloproteinases
col-‐
lagenase
and
elastase.
Both
raw
honey
samples
and
pure
compounds
exerted
moderate
inhib-‐
itory
activity
on
the
tested
enzymes.
More
specifically,
strawberry
tree
and
heather
honey
samples
(100
µg/mL)
inhibited
elastase
62.4%
and
56.1%
and
collagenase
54.3%
and
50.6%,
respectively.
The
isolated
compounds
tested
at
100
µM
exerted
56.7%
and
46.3%
(unedone),
45.2%
and
48.5%
(quercitrin),
38.1%
and
45.7%
(naringenin)
inhibitory
potency
against
elastase
and
collagenase,
respectively.
The
abscisic
acid
isomers
didn’t
appear
to
be
potent
inhibitors
against
both
enzymes.
Spectrophotometric
methods
were
used
to
determine
the
enzymatic
activity.
Elastase
and
collagenase
are
implicated
in
the
wound
healing
process
and
their
overexpression
and
activation
is
thought
to
be
involved
in
the
pathogenesis
of
chronic
wounds.
Improving
basic
knowledge
in
this
area
ultimately
may
help
clarify
the
role
of
honey
in
the
wound
healing
mechanism
and
proactively
intervene
in
an
effort
to
prevent
normal
wounds
from
becoming
chronic
[3].
Keywords:
Honey,
HPTLC,
MPLC,
isolation,
matrix
metalloproteinases
inhibition,
wound
healing
References:
[1] Jull
AB,
Walker
N,
Deshpande
S.
Honey
as
a
topical
treatment
for
wounds.
Cochrane
Data-‐
base
Syst
Rev
2013;
CD005083
[2] Majtan
J.
Honey:
An
immunomodulator
in
wound
healing,
Wound
Rep
Reg
2014;
22:187-‐
192
[3] Armstrong
DG.
The
role
of
Matrix
Metalloproteinases
in
Wound
Healing.
J
Am
Podiatr
Med
Assoc
2002;
92:
12-‐18
P500
Studies
on
the
chemical
stability
of
umbelliprenin,
the
active
principle
of
Ferula
spp.
Vito
Alessandro
Taddeo1,
Francesco
Epifano1,
Philippe
de
Medina2,
Serena
Fiorito1,
Salvatore
Genovese1
1
Department
of
Pharmacy,
University
“G.
d’Annunzio”
of
Chieti-‐Pescara,
Via
dei
Vestini
31,
66100
Chieti
Scalo (CH), Italy, 2 Affichem S.A., Rue de Saint Joseph 9, 31400 Toulouse, France
7-‐Oxyprenylated
coumarins
are
among
the
most
notable
examples
of
biologically
active
oxy-‐
prenylated
plant
secondary
metabolites
occurring
in
nature,
mainly
in
plants
belonging
to
Rutaceae
and
Apiaceae
families.
Umbelliprenin
(UMP)
is
a
farnesyloxy-‐
umbelliferon
deriva-‐
tive
that
has
been
found
in
several
edible
fruits
and
vegetables
including
celery,
coriander,
angelica,
lemon,
and
Ferula
spp.
It
has
been
isolated
in
the
60’s
but
only
recently
the
phyto-‐
chemical
and
biological
profile
has
been
detailed
[1].
Umbelliprenin
was
shown
to
exert
valu-‐
able
and
promising
anti-‐cancer,
anti-‐inflammatory,
anti-‐oxidant,
and
anti-‐protozoal
effects.
Despite
its
effectiveness
as
pharmacological
tool,
studies
on
the
chemical
stability
of
this
ac-‐
tive
principle
have
never
been
reported
in
the
literature.
In
this
talk
data
obtained
from
a
simple,
economic,
and
selective
HPLC
method
for
the
forced
degradation
of
umbelliprenin
indicating
its
chemical
stability
upon
exposure
to
oxidation,
heat,
sun,
and
UV
light
will
be
discussed.
Forced
degradation
studies
provide
insights
into
degradation
pathways
and
side-‐
products
formation
from
the
candidate
drug
substance,
help
in
the
elucidation
of
the
structure
of
the
degradation
products,
and
allow
to
better
set-‐up
formulation
and
package.
In
our
stud-‐
ies
a
C-‐18
reversed-‐phase
packed
column,
was
employed
for
the
separation,
and
the
column
was
thermostated
at
25
±
1
°C
using
a
cool
pocket
chiller.
UV
detector
with
excitation
and
emission
wavelength
of
323
nm
was
used
for
determination
of
umbelliprenin.
Empower
v.2
Software
(Waters
Spa,
Milford,
MA,
USA)
was
used
for
data
acquisition
and
elaboration.
The
flow
rate
was
set
at
1.2
mL/min.,
chromatographic
separation
was
carried
out
using
gradient
eluition.
Samples
(20
μL)
were
injected
by
means
of
a
Rheodyne
injector
fitted
with
a
20-‐μL
loop.
The
method
was
validated
for
accuracy,
precision,
reproducibility,
specificity,
robust-‐
ness,
and
detection
and
quantification
limits,
in
accordance
with
ICH
guidelines.
Results
ob-‐
tained
are
summarized
in
Figures
1
(UMP
in
the
solid
state)
and
2
(UMP
inEtOH
solution).
The
protective
effect
of
several
natural
and
synthetic
antioxidants
on
umbelliprenin
degradation
will
be
also
discussed.
Acknowledgements:
Financial
support
to
this
research
from
University
“G.
d’Annunzio”
of
Chieti-‐Pescara
is
gratefully
acknowledged
Keywords: Antioxidant activity, chemical stability, oxyprenylated coumarins, umbelliprenin
Figure
1.
Degradation
of
UMP
in
the
solid
state
Figure
2.
Degradation
of
UMP
in
ethanolic
solution
References:
[1] Shakeri
A,
Iranshahy
M,
Inrashahi
M.
Biological
properties
and
molecular
targets
of
um-‐
belliprenin
–
a
minireview.
J
As
Nat
Prod
Res
2014;
8:
884-‐889
P501
Discovery
of
novel
protein
tyrosine
phosphatase
1B
inhibitors
from
medicinal
plant
resources
Wei
Li,
Tatsunori
Sasaki,
Toshihisa
Onoda,
Koji
Higai,
Kazuo
Koike
Faculty of Pharmaceutical Sciences, Toho University, Miyama 2-‐2-‐1, Funabashi, Chiba 274-‐8510, Japan
Protein
tyrosine
phosphatase
1B
(PTP1B)
is
a
non-‐transmembrane
protein
tyrosine
phospha-‐
tase.
It
is
expressed
ubiquitously
in
the
classical
insulin-‐targeted
tissues
and
plays
critical
roles
in
negatively
regulating
insulin
and
leptin
signaling
cascades.
Pharmacological
studies
have
shown
that
PTP1B
knockout
mice
exhibited
increased
insulin
sensitivity
and
obesity
resistance.
These
findings
have
led
to
an
intense
interest
in
developing
PTP1B
inhibitors
as
potential
therapies
for
diabetes
and
obesity.
Although
chemical
synthesis
have
resulted
in
the
design
of
potent
synthetic
PTP1B
inhibitors,
some
difficulties,
such
as
high
polarity
and
low
enzyme
selectivity,
must
still
be
overcome
for
the
development
of
new
drugs.
Accordingly,
the
use
of
natural
products
has
received
much
attention
as
an
alternative
approach
for
the
dis-‐
covery
of
novel
PTP1B
inhibitors.
We
have
carried
out
a
series
of
researches
to
discover
novel
PTP1B
inhibitors
from
medicinal
plant
resources,
through
approaches
of
screening
of
plant
extracts
and
compounds
libraries,
bioactivity-‐guided
fractionation,
chemical
structure
elucidation
and
biological
evaluations
[1−4].
The
inhibitory
properties
were
investigated
by
enzymatic
kinetic
analysis,
molecular
docking
simulation,
inhibitory
selectivity
against
other
PTPs,
and
cellular
activity
in
the
insu-‐
lin
signal
transduction
pathway.
Herein,
we
report
some
recent
progress,
in
particular,
of
se-‐
lected
compounds
with
significant
PTP1B
inhibitory
activity
for
the
potential
as
lead
com-‐
pounds
in
future
anti-‐diabetes
drug
development.
Acknowledgements:
This
study
was
partially
supported
by
JSPS
KAKENHI
Grant
Numbers
23790025
and
15K08003.
Keywords: Protein tyrosine phosphatase 1B, natural product, diabetes
References:
[1] Li
S,
Li
W,
Wang
Y,
Asada
Y,
Koike
K.
Prenylflavonoids
from
Glycyrrhiza
uralensis
and
their
protein
tyrosine
phosphatase
1B
inhibitory
activities.
Bioorg
Med
Chem
Lett
2010;
20:
5398-‐5401.
[2] Li
W,
Li
S,
Higai
K,
Sasaki
T,
Asada
Y,
Ohshima
S,
Koike
K.
Evaluation
of
licorice
flavonoids
as
protein
tyrosine
phosphatase
1B
inhibitors.
Bioorg
Med
Chem
Lett
2013;
23:
5836-‐
5839
[3] Sasaki
T,
Li
W,
Higai
K,
Koike
K.
Canthinone
alkaloids
are
novel
protein
tyrosine
phospha-‐
tase
1B
inhibitors.
Bioorg
Med
Chem
Lett
2015;
25:
1979-‐1981
[4] Onoda
T,
Li
W,
Sasaki
T,
Miyake
M,
Higai
K,
Koike
K.
Identification
and
evaluation
of
mag-‐
nolol
and
chrysophanol
as
the
principle
protein
tyrosine
phosphatase-‐1B
inhibitory
com-‐
pounds
in
a
Kampo
medicine,
Masiningan.
J
Ethnopharmacol
2016;
186:
84-‐90.
P502
Structure
characterization
and
immunomodulating
effects
of
pol-‐
ysaccharides
isolated
from
Dendrobium
officinale
Wei
Wei1,
Lei
Feng2,
Shao-‐Ping
Nie2,
and
Quan-‐Bin
Han1
1
School
of
Chinese
Medicine,
Hong
Kong
Baptist
University,
Hong
Kong,
China;
2
State
Key
Laboratory
of
Food Science and Technology, Nanchang University, Nanchang, Jiangxi, China
A
crude
polysaccharide
fraction
(cDOP)
has
been
determined
to
be
the
characteristic
marker
of
Dendrobium
officinale
Kimura
&
Migo
(Orchidaceae),
an
expensive
tea
material
in
Asia.
cDOP
was
de-‐starched
(DOP,
90%
yield),
and
separated
into
two
sub-‐fraction
polysaccharides,
DOPa
and
DOPb,
using
anion-‐exchange
chromatography
column.
DOP,
DOPa,
and
DOPb
were
characterized
by
monosaccharide
composition
and
methylation
analyses,
and
spectral
anal-‐
yses
(FT-‐IR
and
1H
and
13C
NMR).
All
of
them
are
composed
of
mannose
and
glucose,
at
simi-‐
lar
ratios;
and
have
a
similar
structure
with
a
backbone
of
1,
4-‐linked
β-‐D-‐mannopyranosyl
and
β-‐D-‐glucopyranosyl
residues.
Significant
differences
were
observed
only
in
their
molecu-‐
lar
weights.
The
molecular
weights
of
DOP,
DOPa,
and
DOPb
are
7.3×105,
8.1×105,
and
6.7×105
Da,
respectively.
DOP,
DOPa,
and
DOPb
enhanced
cell
proliferation,
TNF-‐α
secretion,
and
phagocytosis
of
RAW264.7
cells
in
a
dose-‐dependent
manner
using
MTT
assays,
ELISA,
and
flow
cytometry,
respectively.
They
also
induced
the
proliferation
of
lymphocytes
alone
and
with
mitogens
[1].
DOPa
and
DOPb
are
thus
proven
to
be
major,
active
polysaccharide
mark-‐
ers
of
D.
officinale.
Rhodiola
rosea
extract
has
been
proved
to
be
a
health
product
for
fatigue
resistance,
so
it
was
used
as
positive
control
for
determining
the
anti-‐fatigue
effect
of
DOP.
Mice
dosed
with
DOP
and
R.
rosea
extract
showed
significant
increases
weight-‐loaded
swim-‐
ming
endurance
duration
time
[2].
The
swimming
time
of
mice
fed
with
DOP
is
much
longer
than
that
of
mice
fed
with
R.
rosea
extract.
Thus,
DOP,
which
exceeds
30%
of
the
dry
herb
by
weight,
has
stronger
anti-‐fatigue
activity
than
R.
rosea
extract.
As
such,
DOP
has
significant
potential
for
pharmaceutical
development
into
health
products
to
reduce
fatigue.
Acknowledgements:
This
study
was
supported
by
HKSAR
Innovation
and
Technology
460
Fund
(ITF),
Tier
3,
ITS/311/09,
and
Hong
Kong
Baptist
University
(RC-‐start
up
grant,
MPCF-‐001-‐2014/2015,
and
FRG2/14-‐15/028)
Keywords: Dendrobium officinale, polysaccharides, chemical structure, RAW264.7 cells, lym-‐
phocytes, anti-‐fatigue effects
References:
[1] Matsuzaki
C,
Hayakawa
A,
Matsumoto
K,
Katoh
T,
Yamamoto
K,
Hisa
K.
Exopolysaccha-‐
rides
produced
by
leuconostoc
mesenteroides
strain
NTM048
as
an
immunostimulant
to
enhance
the
mucosal
barrier
and
influence
the
systemic
immune
response.
J
Agric
Food
Chem
2015;
63:
7009−7015
[2] Chen
Y,
Kong
LD,
Xia
X,
Kung
HF,
Zhang
L.
Behavioral
and
biochemical
studies
of
total
fu-‐
rocoumarins
from
seeds
of
Psoralea
corylifolia
in
the
forced
swimming
test
in
mice.
J
Ethnopharmacol
2005;
96:
451−459
P503
SAR
study
and
antitumor
potential
of
plant
natural
product
mili-‐
usanes
and
their
derivatives
Wen-‐Hui
Pan,
Yi-‐Fu
Guan,
Kang-‐Lun
Liu,
Xin-‐Ya
Xu,
Xun
Song,
Dong-‐Ying
Wang,
Wen-‐Jian
Xie,
Siu
Wai
Tsang,
Hong-‐Jie
Zhang
School of Chinese Medicine, Hong Kong Baptist University, Hong Kong SAR, People’s Republic of China
In
our
previous
research,
bioassay-‐directed
fractionation
of
Miliusa
sinensis
Finet
&
Gagnep.
(Annonaceae)
led
to
the
discovery
of
miliusane
anticancer
lead
molecules
[1].
As
a
continuous
study,
we
have
investigated
the
chemical
constituents
of
another
plant
species
(M.
balansae)
in
the
same
genus.
Separation
of
the
dichloromethane
extract
of
this
plant
led
to
the
isolation
of
8
known
miliusanes
including
miliusate
(1)
and
miliusol
(2),
the
potent
antitumor
com-‐
pounds
discovered
in
our
previous
study.
In
this
study,
we
investigated
the
in
vivo
antitumor
activity
of
the
miliusane
compounds
using
the
hollow
fibre
as
well
as
the
xenograft
mouse
models.
The
animal
studies
demonstrated
the
antitumor
potency
of
miliusol
is
comparable
with
that
of
the
clinically
used
drug
paclitaxel,
but
with
lower
toxicity.
In
order
to
determine
the
structure-‐activity
relationship
(SAR)
of
miliusanes,
we
further
synthesized
15
miliusane
derivatives
(3-‐17).
From
the
SAR
study,
we
found
that
although
the
α,
β-‐unsaturated
carbonyl
structure
played
an
important
role
in
cytotoxicity
for
some
miliusanes,
this
functional
group
is
not
essential
for
the
bioactivity
of
miliusanes.
Indeed,
the
two
Michael
addition
products
13
and
14,
which
contained
no
α,
β-‐unsaturated
carbonyl,
showed
equivalent
cancer
cell
killing
activities
(IC50
against
HCT116:
1.88
and
1.83
μM,
respectively)
as
those
of
miliusate
and
mili-‐
usol.
An
α,
β-‐unsaturated
carbonyl
structure
has
been
known
to
contribute
to
the
biological
activities
of
many
compounds.
However,
the
same
functional
group
may
also
cause
severe
side
effects
because
of
its
reactivity
with
many
protein
receptors.
By
eliminating
the
function-‐
al
group
while
retain
the
bioactivity,
compounds
such
as
13
and
14
may
have
less
toxicity
in
comparison
with
miliusol
and
miliusate.
This
SAR
study
will
provide
us
valuable
information
for
rational
design
to
synthesize
miliusane
analogs
as
druggable
candidates
for
anticancer
treatment.
Acknowledgements:
The
work
described
in
this
paper
was
supported
by
the
Research
Grants
Council
of
the
Hong
Kong
Special
Administrative
Region,
China
(Project
No.
HKBU
12103014)
and
HKBU
Interdisci-‐
plinary
Research
Matching
Scheme
(RC-‐IRMS/12-‐13/03).
Keywords: Miliusane, Miliusa balansae, isolation and identification, structure modification,
antitumor activity, in vivo activity
References:
[1] Zhang
HJ,
Ma
CY,
Hung
VN
et
al.
Miliusanes,
A
class
of
cytotoxic
agent
from
Miliusa
sinensis.
J
Med
Chem
2006;
49:
693-‐708
P504
Cytotoxicity
effects
and
apoptosis
induction
by
cycleanine
and
tetrandrine
Fidelia
I.
Uche1,2,
Falko
Drijfhout3,
James
McCullagh4,
Alan
Richardson1,
Wen-‐Wu
Li1
1
Institute
for
Science
and
Technology
in
Medicine,
Keele
University,
UK,
2
Faculty
of
Pharmaceutical
Sci-‐
ences,
University
of
Port
Harcourt,
Nigeria,
3
Chemical
Sciences
Research
Centre,
Keele
University,
UK,
4
Chemical
Research
Laboratory,
University
of
Oxford,
UK
Ovarian
cancer
remains
one
of
the
main
causes
of
death
in
all
gynecologic
malignancies
[1].
Natural
products
continue
to
be
important
sources
of
clinically
approved
anti-‐cancer
drugs
[2,
3].
Triclisia
subcordata
Oliv
(Menispermeaceae)
is
a
medicinal
plant
traditionally
used
for
the
treatment
of
various
diseases
[4],
including
cancer,
in
West
Africa.
This
study
aims
to
evaluate
the
in
vitro
anti-‐ovarian
cancer
activities
of
the
crude
extracts
and
the
isolated
components
in
T.
subcordata.
The
ethanol
extract
of
T.
subcordata
and
its
fractions
(crude
alkaloids)
were
screened
for
in
vitro
anti-‐ovarian
cancer
activities
on
Ovcar-‐8,
Ovcar-‐4,
A2780,
and
Igrov-‐1
ovarian
cancer
cell
lines
using
the
Sulforhodamine
B
assay
method
to
measure
cell
growth.
Bioassay-‐guided
fractionation
using
silica
gel
column
chromatography
and
HPLC
were
used
to
isolate
the
bioactive
compound,
whose
identity
and
structure
was
identified
by
NMR
and
LC-‐
MS
techniques.
Caspase
and
PARP
cleavage
assays
were
used
to
detect
apoptotic
activities.
The
effect
of
isolated
pure
compounds
on
cell
cycle
and
apoptosis
was
analyzed
by
flow
cy-‐
tometry.
Results:
The
crude
alkaloids
showed
the
strongest
activity
in
cell
growth
assays
on
A2780
and
Ovcar-‐8
cell
lines
(IC50
<
2.4
µg/mL).
A
bisbenzylisoquinoline
alkaloid-‐cycleanine
was
isolated
using
HPLC
and
identified
by
MS
and
NMR
analyses.
The
IC50values
of
cycleanine
and
tetrandrine
ranged
from
7
to
14
µM
on
A2780,
Ovcar-‐8,
Ovcar-‐4
and
Igrov-‐1
ovarian
can-‐
cer
cell
lines.
The
IC50
of
cycleanine
on
human
normal
ovarian
surface
epithelial
cells
was
35
±
1
µM
hinting
at
modest
selectivity
towards
cancer
cells.
Both
cycleanine
and
tetrandrine
caused
apoptosis
as
shown
by
activation
of
caspases
3/7
and
cleavage
of
poly
(ADP)
ribose
polymerase
(PARP)
to
form
PARP-‐I.
The
percentage
of
Ovcar-‐8
cells
in
subG1
phase
increased
after
exposure
of
cycleanine
and
tetrandrine
to
cells
for
48h
compared
to
control.
In
conclu-‐
sion,
cycleanine,
like
its
isomer
–
tetrandrine
isolated
from
T.
subcordata,
could
be
a
potential
new
anti-‐ovarian
cancer
agent
acting
through
the
apoptosis
pathway.
Acknowledgements:
We
thank
Nigerian
ETF
and
NDDC
for
funding.
We
also
thank
Mr.
John
Clews
for
assistance
in
NMR
measurements.
Keywords: Triclisia subcordata, cycleanine, tetrandrine, anti-‐proliferation, apoptosis, ovarian
cancer
References:
[1] Siegel
R,
Naishadham
D,
Jemal
A.Cancer
statistics.
CA
Cancer
J
Clin
2013;
63:
11−30
[2] Li
WW,
Johnson-‐Ajinwo
OR,
Uche
F.
Potential
of
Phytochemicals
and
their
Derivatives
in
the
Treatment
of
Ovarian
Cancer.
In:
Collier
BR,
editor.
Handbook
on
Ovarian
Cancer:
Risk
Factors,
Therapies
and
Prognosis.
Nova
Science
publishers,
USA,
2015;
155−180
[3] Johnson-‐Ajinwo
OR,
Richardson
A,
Li
WW.
Cytotoxic
effects
of
stem
bark
extracts
and
pure
compounds
from
Margaritaria
discoidea
on
human
ovarian
cancer
cell
lines.
Phytomedi-‐
cine
2015;
22:
1−4
[4] Abo
KA,
Lawal
IO,
Ogunkanmi
A.
Evaluation
of
extracts
of
Triclisia
subcordata
Oliv
and
Heinsia
crinita
(Afz)
G.
Taylor
for
antimicrobial
activity
against
some
clinical
bacterial
iso-‐
lates
and
fungi.
Afr
J
Pharm
Pharmacol
2011;
5:
125−131
P505
Antibacterial
effects
of
the
essential
oil
from
flower
buds
of
Mag-‐
nolia
biondii
Pamp
Omar
Aldulaimi1,
2,
Wen-‐Wu
Li1
1
Institute
for
Science
and
Technology
in
Medicine,
Keele
University,
Stoke-‐on-‐Trent,
Thornburrow
Drive,
ST4 7QB, UK, 2 College of Pharmacy, Al-‐Mustansiriyah University, Iraq
Flower buds of Magnolia biondii Pamp (family Magnoliaceae) is known as Xin-‐Yi in the Chi-‐
nese Pharmacopoeia, and is widely used for the treatment of allergic rhinitis, rhinosinusitis,
nasal congestion, and headache.1 Bacterial infection caused by Staphylococcus aureus in the-‐
nasal-‐sinus mucosa is one of key factors which could cause rhinosinusitis,2 thus it is important
to evaluate the antibacterial effect of the extracts from M. biondii, which may provide scientific
evidence of using Xin-‐Yi for the treatment of rhinosinusitis. In this study, the essential oil and
lignan-‐rich extract isolated from the flower buds of Magnolia biondii Pamp (Xin-‐Yi) were in-‐
vestigated for their chemical compositions and in vitro antibacterial activities. GC-‐MS analysis
of the Magnolia essential oil disclosed the presence of 56 compounds including camphor (10.6
%), eucalyptol (25.0 %), linalool (5.8 %), terpine-‐4-‐ol (8.4 %), alpha-‐terpineol (19.8 %), al-‐
pha-‐cadinol (3.3 %), citronellol (2.9 %), geraniol (2.3 %), and trans-‐farnesol (8.7 %). Both GC-‐
MS and NMR analyses of the chloroform extract disclosed the presence of 7 tetrahydrofurofu-‐
ran lignans that were demethoxyaschantin, fargesin, epieudesmin, eudesmin, aschantin, mag-‐
nolin, and yangambin. The essential oil showed stronger antibacterial activities than the
lignan-‐rich extract against five bacteria including pathogenic Staphylococcus aureus and S.
epidermidis with MICs ranging from 250 to 500 µg ml-‐1 using microplate Alamar blue assay.
Time-‐kill kinetics was used to monitor the survival characteristics of S. aureus and Escherichia
coli in the presence of the essential oil over 24 hours, which indicated rapidly bactericidal ef-‐
fects. Scanning electron microscopy (Figure 1) showed the change of morphological appear-‐
ance of S. aureus through destruction its cell wall and membrane by the Magnolia oil.
Figure
1.
Scanning
electron
micrograph
of
control
S.
aureus
at
1
hour
of
incubation
at
37°C
(A).
Scan-‐
ning
electron
micrograph
of
S.
aureus
after
exposure
to
Magnolia
oil
(2
MIC)
for
1h
(B).
Scanning
elec-‐
tron
micrograph
of
S.
aureus
after
exposure
to
Magnolia
oil
(2
MIC)
for
24h
(C).
Arrows
show
lysis
of
cells
and
leakage
of
cellular
contents.
Acknowledgements:
This
work
was
supported
by
MOHSER:
Ministry
Of
Higher
Education
and
Scientific
Research/IRAQ
(A
PhD
studentship
to
Omar
Aldulaimi).
We
are
grateful
to
Nigel
Bowers
and
Prof.
Paul
Horrocks
for
their
assistance
in
antimicrobial
assay,
and
Karen
Walker
and
Prof.
David
Furness
for
their
help
with
SEM
Keywords:
Magnolia
biondii
Pamp,
essential
oils,
GC-‐MS,
time-‐kill
assay,
scanning
electron
microscopy,
Staphylococcus
aureus
References:
[1] Shen
Y,
Li
CG,
Zhou
SF,
Pang
EC,
Story
DF
and
Xue
CC.
Chemistry
and
bioactivity
of
Flos
Magnoliae,
a
Chinese
herb
for
rhinitis
and
sinusitis.
Cur
Med
Chem
2008;
15:
1616-‐1627
[2] Redinbo
MR.
The
microbiota,
chemical
symbiosis,
and
human
disease.
J
Mol
Biol
2014;
426:
3877-‐3891
P506
Antimicrobial
and
antioxidant
efficacy
of
Acokanthera
oblongifo-‐
lia
(Apocynaceae)
used
for
skin
diseases
by
Xhosa
tribe
of
Amathole
District,
Eastern
Cape,
South
Africa
Wilfred
M.
Otang1,
Anthony
J.
Afolayan2
1
IKS
Centre,
Faculty
of
Science
and
Agriculture,
North
West
University,
South
Africa;
2
Medicinal
Plants
and
Economic
Development
(MPED)
Research
Centre,
Department
of
Botany,
University
of
Fort
Hare,
Private
Bag
X1314,
Alice
5700,
South
Africa
Acokanthera
oblongifolia
Hochst.
(Apocynaceae)
is
an
evergreen
medicinal
shrub
used
for
snakebites,
itches,
wounds
and
internal
worms
and
the
relief
of
itchy
conditions
and
other
skin
disorders
by
the
Mpondo
and
Xhosa
tribes
in
South
Africa
[1].
The
objective
of
this
study
was
to
investigate
the
efficacy
of
the
plant
extracts
against
selected
pathogens
that
cause
hu-‐
man
skin
disorders
and
evaluation
of
the
antioxidant
capability
for
validation
of
folk
uses
of
the
plant.
The
agar
diffusion
and
micro-‐dilution
methods
[2]
were
used
to
determine
the
an-‐
timicrobial
activities
of
the
extracts
against
selected
bacteria
and
fungi.
The
highest
antibacte-‐
rial
activity
(inhibition
zone
diameter
>
19mm)
was
obtained
with
the
acetone
extract
against
Pseudomonas
aeruginosa,
Shigella
sonnei,
Shigella
flexneri,
Bacillus
cereus,
Streptococcus
pyo-‐
gens
and
Bacillus
subtilis.
None
of
the
extracts
were
active
against
the
tested
fungi,
apart
from
the
acetone
extract
which
showed
strong
inhibitory
activity
against
Candida
glabrata.
The
ABTS
scavenging
activity
of
the
ethanol
extract
was
higher
than
that
of
the
acetone
extract,
BHT
and
Gallic
acid,
except
at
concentrations
greater
than
0.15
mg/ml.
The
sensitivity
shown
by
these
pathogenic
organisms
especially
towards
the
acetone
extract
of
A.
oblongifolia
may
partially
justify
the
ethnomedicinal
use
of
the
plant.
Acknowledgements:
National
Research
Foundation
(NRF)
and
Govan
Mbeki
Research
and
Development
Centre
(GMRDC)
of
the
University
of
Fort
Hare,
South
Africa
are
highly
acknowledged
Key
words:
Antimicrobial,
antioxidant,
Acokanthera
oblongifolia,
skin
diseases,
Eastern
Cape,
South
Africa
References:
[1] Hassan
R,
Hassan
EM,
Ibrahim
NA,
Nazif
NM.
Triterpenes
and
cytotoxic
activity
of
Acokan-‐
thera
oblongifolia
Hochst
Growing
in
Egypt.
Res
J
Pharm
Biol
Chem
Sci
2015;
1677−1686
[2] Nesy
EA,
Mathew
L.
Studies
on
antimicrobial
and
antioxidant
efficacy
of
Thevetia
neriifo-‐
lia,
Juss
leaf
extracts
against
human
skin
pathogens.
Int
J
Pharm
Sci
Drug
Res
2014;
6:
164−168
P507
Chemical
constituents
from
Melicope
pteleifolia
leaves
and
their
anti-‐influenza
virus
activities
Ngoc-‐Hieu
Nguyen,
Thi-‐Kim-‐Quy
Ha,
Won
Keun
Oh
Korea
Bioactive
Natural
Material
Bank,
Research
Institute
of
Pharmaceutical
Sciences,
College
of
Phar-‐
macy,
Seoul
National
University,
Seoul
08826,
Republic
of
Korea
Melicope
pteleifolia
(Champ.
Ex
Benth)
T.G.
Hartley
(synonym
Euodia
lepta
(Spreng.)
Merr.),
belonging
to
Rutaceae
family,
is
a
medium-‐sized
tree,
which
is
widely
distributed
in
South
and
South-‐East
Asia.
In
traditional
medicine,
the
extract
prepared
from
M.
pteleifolia
is
empirically
used
for
the
treatment
of
remittent
fever,
colds
and
various
inflammatory
conditions.
Previ-‐
ous
phytochemical
investigations
revealed
that
the
main
constituents
of
this
plant
were
ben-‐
zopyrans
[1],
furoquinoiline
alkaloids
[2],
and
glycosidic
compounds
[3].
Our
ongoing
chemi-‐
cal
investigation
revealed
the
occurrence
of
rare
spiroketal
constitutents
from
the
polar
frac-‐
tions
of
a
methanol
extract
of
M.
pteleifolia
leaves’.
The
phytochemical
investigation
was
car-‐
ried
out
by
employing
various
chromatographic
methods,
resulting
in
the
isolation
of
a
total
24
compounds
(1-‐24).
On
the
basis
of
spectroscopic
analyses,
including
NMR,
UV,
IR
and
MS
and
by
spectral
comparison
with
published
literature,
the
isolated
compounds
were
deter-‐
mined
as
five
new
acetophenones
bearing
spiroketal-‐hexofuranoside
rings
(1-‐5),
one
new
C-‐
glycosidic
acetophenone
(11)
and
two
new
benzopyrans
(13,
14),
together
with
16
known
compounds.
This
is
also
the
first
report
of
natural
occurrence
of
spiroketal-‐hexofuranosides
(1-‐5)
from
M.
pteleifolia
leaves.
To
scientifically
clarify
the
traditional
usage
of
M.
pteleifolia
leaves
for
treating
fever
and
colds,
all
isolated
compounds
were
screened
for
their
inhibitory
activities
against
neuraminidases
from
four
different
influenza
viral
strains
(H1N1,
H9N2,
wild-‐type
H1N1
and
oseltamivir-‐resistant
H1N1
with
H274Y
mutation)
[4].
Among
them,
compound
10
was
found
to
exhibit
the
strongest
inhibition
against
those
types
of
neuramini-‐
dase
(IC50
=
24.93
±
3.46,
23.19
±
5.41,
26.67
±
5.16
and
40.16
±
4.50
µM,
respectively).
Fur-‐
thermore,
in
the
cytopathic
effect
(CPE)
assay,
compounds
7,
8
and
10
also
showed
the
reduc-‐
tion
of
H1N1-‐induced
cytopathic
effect
in
MDCK
cells.
Keywords: Melicope pteleifolia, Rutaceae, spiroketal-‐hexofuranosides, spiroketal, benzopy-‐
ran, neuraminidase, oseltamivir-‐resistant, H1N1.
References:
[1] Kamperdick
C,
Van
NH,
Van
Sung
T,
Adam
G.
Benzopyrans
from
Melicope
ptelefolia
leaves.
Phytochemistry
1997;
45:
1049-‐1056
[2] Sichaem
J,
Jirasirichote
A,
Sapasuntikul
K,
Khumkratok
S,
Sawasdee
P,
Do
TM,
Tip-‐Pyang
S.
New
furoquinoline
alkaloids
from
the
leaves
of
Evodia
lepta.
Fitoterapia
2014;
92:
270-‐
273
[3] Zhang
Y,
Yang
LJ,
Jiang
K,
Tan
CH,
Tan
JJ,
Yang
PM,
Zhu
DY.
Glycosidic
constituents
from
the
roots
and
rhizomes
of
Melicope
pteleifolia.
Carbohyd
Res
2012;
361:
114-‐119
[4] Dao
TT,
Tung
BT,
Nguyen
PH,
Thuong
PT,
Yoo
SS,
Kim
EH,
Kim
SK,
Oh
WK.
C-‐Methylated
flavonoids
from
Cleistocalyx
operculatus
and
their
inhibitory
effects
on
novel
influenza
A
(H1N1)
neuraminidase.
J
Nat
Prod
2010;
73:
1636-‐1642
P508
Antitumor
activity
of
periplocin,
isolated
from
Telectadium
dong-‐
naiense,
in
colorectal
cancer
cells
via
suppression
of
Wnt/β-‐
catenin
signaling
Won
Kyung
Kim1,
Ba-‐duc
Hiep1,
Tae
Joon
Choi1,
Je
Do
Oh1,
Hyen
Joo
Park1,
Sei
Ryang
Oh2,
Sang
Kook
Lee1
1College
of
Pharmacy,
Seoul
National
University,
1
Gwanak-‐ro
Gwanak-‐gu,
08826
Seoul,
Korea,
2Korea
Research
Institute,
Bioscience
and
Biotechnology
International
Biological
Material
Research
Center,
125
Gwahak-‐ro
Yousung-‐gu,
34141
Daejeon,
Korea
In
the
early
events
of
carcinogenesis,
abnormal
activation
of
Wnt/β-‐catenin
and
up-‐regulation
of
β-‐catenin/T-‐cell
factor
(TCF)
responsive
transcription
play
a
critical
role,
especially
in
colo-‐
rectal
cancers
[1].
On
this
line,
we
examined
the
effects
of
14,000
plant
extracts
on
Wnt/β-‐
catenin
signaling.
A
reporter
gene
assay
system
was
employed
which
HEK293
human
embry-‐
onic
kidney
cells
were
transiently
transfected
with
β-‐catenin
and
TCF4
plasmids,
and
TOPflash
activity
was
determined.
Among
the
plant
extract,
the
extracts
of
Telectadium
dong-‐
naiense
Pierre
ex
Costantin
(TDPC,
IC50=2.1
μg/ml),
exhibited
the
most
potent
inhibitory
activ-‐
ity
in
TOPflash
assay
without
cytotoxicity.
Three
main
compounds
(periplocin,
rutin,
and
1,4-‐
O-‐dicaffeoylquinic
acid)
were
isolated
and
purified
from
TDPC
with
liquid
chromatography,
mass
spectrometry,
and
nuclear
magnetic
resonance.
Among
the
compounds,
periplocin
showed
the
largest
cytotoxic
effect
on
cancer
cells
and
inhibitory
effect
on
Topflash
activity.
Further
investigation
of
periplocin
on
the
regulation
of
Wnt/β-‐catenin
signaling
revealed
that
periplocin
inhibited
nuclear
translocation
of
β-‐catenin
and
subsequently
expressions
of
target
genes
such
as
c-‐myc,
cyclin
D1,
and
survivin
were
significantly
suppressed.
In
addition,
perip-‐
locin
also
exhibited
in
vitro
anti-‐proliferative
effects
and
in
vivo
antitumor
activity
in
a
tumor
xenograft
model
using
nude
mice
implanted
with
HCT116
colon
cancer
cells.
Ex
vivo
biochem-‐
ical
analysis
of
tumors
from
nude
mouse
xenografts
showed
suppression
of
Wnt
target
genes
by
periplocin.
These
findings
implicate
the
involvement
of
the
Wnt/β-‐catenin
signaling
path-‐
way
in
the
antitumor
activity
of
periplocin.
Acknowledgements:
This
work
was
supported
by
the
National
Research
Foundation
of
Korea
grant
(NRF-‐
2012K1A1A3307877).
Keywords: Telectadium dongnaiense, Wnt/β-‐catenin signaling, human colon cancer, periplo-‐
cin
References:
[1] MacDonald
BT,
Tamai
K,
He
X.
Wnt/beta-‐catenin
signaling:
components,
mechanisms,
and
diseases.
Dev
Cell
2009;
17:
9−26
P509
Cytotoxic
triterpenes
from
the
twigs
of
Chaenomeles
sinensis
Won
Se
Suh1,
Chung
Sub
Kim1,
Kyoung
Jin
Park1,
Joon
Min
Cha1,
Oh
Kil
Kwon1,
Sang
Un
Choi2,
Kang
Ro
Lee1
1Natural
Products
Laboratory,
School
of
Pharmacy,
Sungkyunkwan
University,
Suwon
16419,
Republic
of
Korea,
2Korea
Research
Institute
of
Chemical
Technology,
Daejeon
34114,
Republic
of
Korea
Chaenomeles
sinensis
Koehne,
belonging
to
the
Rosaceae
family,
is
widely
distributed
in
Korea,
Japan,
and
China.
The
fruits
of
this
woody
tree
have
long
been
used
in
Korean
traditional
med-‐
icine
for
the
treatment
of
various
disorders
such
as
cough,
the
common
cold,
diarrhea,
in-‐
flammatory
diseases,
and
dry
beriberi,
as
well
as
to
make
tea,
jam,
and
liquor
[1-‐3].
Several
triterpenes,
flavonoids,
lignans,
sesquiterpenes,
and
simple
phenolic
compounds
have
been
isolated
from
C.
sinensis,
and
some
of
these
compounds
and
the
extract
of
C.
sinensis
have
been
shown
to
display
antioxidant,
antiviral,
antidiabetic,
antiacetylcholinesterase,
antihypergly-‐
cemic,
and
antihyperlipidemic
activities
[3,
4].
In
the
process
of
searching
for
bioactive
com-‐
pounds
from
this
source,
we
further
isolated
three
new
triterpenes
(1-‐3),
along
with
20
known
ones
(4-‐23).
The
chemical
structures
of
the
isolated
compounds
were
elucidated
by
extensive
NMR
data
(1H
and
13C
NMR,
1H-‐1H
COSY,
HSQC,
HMBC,
and
NOESY),
HRMS,
and
chemical
reaction.
All
the
isolated
compounds
(1-‐23)
were
evaluated
for
their
cytotoxic
activ-‐
ity
against
A549,
SK‑OV‑3,
SK‑MEL‑2,
and
HCT15
cell
lines
in
vitro
using
the
SRB
bioassay
and
among
them,
compounds
3,
17,
20,
and
23
showed
potent
cytotoxic
activitiy
(IC50
0.92-‐
3.09 μM).
Chaenomelin
(1)
3β-‐O-‐(cis-‐Ferulyl)-‐2α,19α-‐dihydroxy-‐urs-‐12-‐en-‐28-‐oic
acid
(2)
Betulin-‐3β-‐cis-‐caffeate
(3)
Acknowledgements:
This
research
was
supported
by
the
Basic
Science
Research
Program
through
the
National
Research
Foundation
of
Korea
(NRF)
funded
by
the
Ministry
of
Education,
Science
and
Technol-‐
ogy
(2013R1A1A2A10005315).
Keywords: Chaenomeles sinensis, Rosaceae, triterpene, cytotoxicity
References:
[1] Lee
MH,
Son
YK,
Han
YN.
Tissue
factor
inhibitory
flavonoids
from
the
fruits
of
Chaeno-‐
meles
sinensis.
Arch
Pharm
Res
2002;
25:
842−850
[2] Kim
HK,
Jeon
WK,
Ko
BS.
Flavanone
glycoside
from
the
fruits
of
Chaenomeles
sinensis.
Nat
Prod
Sci
2000;
6:
79−81
[3] Kim
CS,
Kwon
OW,
Kim
SY,
Choi
SU,
Kim
KH,
Lee
KR.
Five
new
oxylipins
from
Chaenomeles
sinensis.
Lipids
2014;
49:
1151−1159
[4] Sawai-‐Kuroda
R,
Kikuchi
S,
Shimizu
YK,
Sasaki
Y,
Kuroda
K,
Tanaka
T,
Yamamoto
T,
Sakurai
K,
Shimizu,
K.
A
polyphenol-‐rich
extract
from
Chaenomeles
sinensis
(Chinese
quince)
in-‐
hibits
influenza
A
virus
infection
by
preventing
primary
transcription
in
vitro.
J
Eth-‐
nopharmacol
2013;
146:
866−872
P510
Four
new
flavonoid
glycosides
from
the
white
flower
of
Impatiens
balsamina
Won
Se
Suh1,
Chung
Sub
Kim1,
Kyoung
Jin
Park1,
Joon
Min
Cha1,
Oh
Kil
Kwon1,
Sang
Zin
Choi2,
Mi
Won
Son2,
Kang
Ro
Lee1
1Natural
Products
Laboratory,
School
of
Pharmacy,
Sungkyunkwan
University,
Suwon
16419,
Republic
of
Korea, 2Dong-‐A ST Research Institute, Kiheung, Yongin 17073, Republic of Korea
Impatiens
balsamina
L.,
also
known
as
Garden
balsam
or
Rose
balsam,
is
an
annual
herba-‐
ceous
plant
of
the
Balsamineaceae
family
grown
as
an
ornamental
garden
plant.
I.
balsamina
has
been
used
in
traditional
Chinese
medicine,
and
in
some
areas
of
China
is
used
as
a
vegeta-‐
ble
or
anticancer
herb
[1].
The
aerial
parts
of
I.
balsamina
have
been
used
for
the
treatment
of
articular
rheumatism,
bruises,
and
beriberi
[2],
whereas
the
seeds
have
been
used
to
treat
lumps,
puerperal
pain,
and
cancer
[3].
The
flower
of
this
plant
also
has
been
used
to
treat
dermititis,
lumbago,
neuralgia,
burns,
and
scalds
[2].
Previous
studies
have
suggested
that
flavonoids
and
naphthoquinones
from
the
flower
of
I.
balsamina
are
associated
with
antipru-‐
ritic,
antianaphylactic,
and
anti-‐inflammatory
properties
[4-‐6].
As
part
of
the
search
for
bioac-‐
tive
constituents
of
Korean
medicinal
plants,
we
investigated
the
active
constituents
of
I.
bal-‐
samina.
In
the
present
study,
we
isolated
four
new
flavonoid
glycosides,
Balsamiside
A-‐D
(1-‐
4)
and
nine
known
compounds
(5-‐13),
from
the
white
flower
of
I.
balsamina.
The
structures
of
the
new
compounds
(1-‐4)
were
elucidated
by
spectroscopic
data
analysis
including
1D
and
2D
NMR
(1H
and
13C
NMR,
1H-‐1H
COSY,
HSQC,
and
HMBC),
ECD
calculation
and
chemical
methods.
Study
on
anti-‐inflammatory
and
neuroprotective
effect
of
isolates
(1-‐13)
are
in
pro-‐
gress.
Balsamiside
A
(1)
Balsamiside
B
(2)
Balsamiside
C
(3)
Acknowledgements:
This
study
was
supported
by
a
research
fund
from
the
Dong-‐A
ST
Research
Center,
2015.
Keywords: Impatiens balsamina, Balsamineaceae, flavonoid glycosides, ECD calculation
References:
[1] Su
BL,
Zeng
R,
Chen
JY,
Chen
CY,
Guo
JH,
Huang
CG.
Antioxidant
and
antimicrobial
proper-‐
ties
of
various
solvent
extracts
from
Impatiens
balsamina
L.
stems.
J
Food
Sci
2012;
77:
614-‐619
[2] Imam
MZ,
Nahar
N,
Akter
S,
Rana
MdS.
Antinociceptive
activity
of
methanol
extract
of
flowers
of
Impatiens
balsamina.
J
Ethnopharmacol
2012;
142:
804-‐810
[3] Lei
J,
Qian
SH,
Jiang
JQ.
Two
new
flavone
glycosides
from
the
seeds
of
Impatiens
balsamina
L.
J
Asian
Nat
Prod
Res
2010;
12:
1033-‐1037
[4] Fukumoto
H,
Yamaki
M,
Isoi
K,
Ishiguro
K.
Antianaphylactic
effects
of
the
principal
com-‐
pounds
from
the
white
petals
of
Impatiens
balsamina
L.
Phytother
Res
1996;
10:
202-‐206
[5] Ishiguro
K,
Oku
H.
Antipruritic
effect
of
flavonol
and
1,4-‐naphthoquinone
derivatives
from
Impatiens
balsamina
L.
Phytother
Res
1997;
11:
343-‐347
[6] Oku
H,
Ishiguro
K.
Cyclooxygenase-‐2
inhibitory
1,4-‐naphthoquinones
from
Impatiens
bal-‐
samina
L.
Biol
Pharm
Bull
2002;
25:
658-‐660
P511
Bioactive
chemical
constituents
from
the
twigs
of
Salix
glandu-‐
losa
Won
Se
Suh,
Chung
Sub
Kim,
Kyoung
Jin
Park,
Joon
Min
Cha,
Dong
Hyun
Kim,
Kang
Ro
Lee
Natural
Products
Laboratory,
School
of
Pharmacy,
Sungkyunkwan
University,
Suwon
16419,
Republic
of
Korea
Salix
glandulosa
Seemen
(=
S.
chaenomeloides
Kimura,
Salicaceae),
also
known
as
“Korean
King
Willow”,
is
an
indigenous
plant
in
East
Asia.
The
genus
Salix
is
known
as
a
important
me-‐
dicinal
herbs,
since
salicin,
a
precursor
of
acetylsalicylic
acid
(aspirin),
flavonoids,
terpenoids,
lignans
and
phenolic
compounds
with
biological
activities
have
been
reported
from
these
sources
[1-‐4].
Especially,
salicin
and
salicylic
acid
have
important
pharmacological
effects
on
treatment
of
fever,
pain,
and
inflammation,
whereas
acetylsalicylic
acid
(aspirin),
having
simi-‐
lar
activities,
is
a
less
toxic
compound
[5].
However,
only
few
studies
on
phytochemical
and
biological
investigations
on
S.
glandulosa
have
been
reported
previously.
The
presence
of
sev-‐
eral
constituents,
including
salicin
derivatives
and
flavonoids,
were
reported
from
S.
glandu-‐
losa.6
In
the
process
of
finding
new
bioactive
compounds
from
Korean
medicinal
plants,
we
isolated
a
new
phenolic
compound
(1),
along
with
22
known
ones
(2-‐23)
from
the
twigs
of
S.
glandulosa.
The
chemical
structure
of
the
new
compound
(1)
was
determined
by
NMR
(1H
and
13C
NMR,
1H-‐1H
COSY,
HSQC,
HMBC,
and
NOESY),
HRMS,
and
CD
data.
All
isolated
compounds
(1–23)
were
evaluated
for
their
cytotoxicity
against
four
human
tumor
cell
lines
(A549,
SK-‐
OV-‐3,
SK-‐MEL-‐2,
and
HCT15),
for
their
potential
neuroprotective
effects
through
induction
of
nerve
growth
factor
(NGF)
in
C6
glioma
cells,
and
for
their
effects
on
nitric
oxide
(NO)
levels
in
lipopolysaccharide
(LPS)-‐stimulated
murine
microglia
BV2
cells.
Acknowledgements:
This
research
was
supported
by
the
Basic
Science
Research
Program
through
the
National
Research
Foundation
of
Korea
(NRF)
funded
by
the
Ministry
of
Education,
Science
and
Technol-‐
ogy
(2012R1A5A2A28671860).
Keywords: Abies holophylla, Pinaceae, lignan, cytotoxicity, nitric oxide
References:
[1] Evans
TP,
Clausen
TP,
Reichardt
PB.
Structurally
intriguing
glucosides
from
Alaskan
Little-‐
tree
Willow
(Salix
arbusculoides).
J
Nat
Prod
1995;
58:
1897-‐1900
[2] Hsu
FL,
Nonaka
GI,
Nishioka
I.
Acylated
flavanols
and
procyanidins
from
Salix
sieboldiana.
Phytochemistry
1985;
24:
2089-‐2092
[3] Yang
H,
Lee
SH,
Sung
SH,
Kim
J,
Kim
YC.
Neuroprotective
compounds
from
Salix
pseudo-‐
lasiogyne
twigs
and
their
anti-‐amnesic
effects
on
scopolamine-‐induced
memory
deficit
in
mice.
Planta
Med
2013;
78:
78-‐82
[4] Tantry
MA,
Shah
S,
Dar
MY,
Mir
MM,
Ghazanfar
K,
Sheikh
FA,
Khuroo
MA,
Akbar
S.
9,10-‐
seco-‐9,19-‐cyclolanostane
triterpene
from
Salix
caprea
L.
(Goat
Willow).
Nat
Prod
Res
2013;
27:
171-‐175
[5] Madan
RK,
Levitt
J.
A
review
of
toxicity
from
topical
salicylic
acid
preparations.
J
Am
Acad
Dermatol
2014;
70:
788-‐792
[6] Mizuno
M,
Kato
M,
Misu
C,
Iinuma
M,
Tanaka
T.
Chaenomeloidin:
a
phenolic
glucoside
from
leaves
of
Salix
chaenomeloides.
J
Nat
Prod
1991;
54:
1447-‐1450
P512
Fatty
acids
from
the
stem
bark
of
Sorbus
commixta
and
their
bio-‐
logical
activity
Won
Se
Suh1,
Chung
Sub
Kim1,
Kyoung
Jin
Park1,
Joon
Min
Cha1,
Tae
Hyun
Lee1,
Sun
Yeou
Kim2,3,
Kang
Ro
Lee1
1Natural
Products
Laboratory,
School
of
Pharmacy,
Sungkyunkwan
University,
Suwon
16419,
Republic
of
Korea,
2Gachon
Institute
of
Pharmaceutical
Science,
Gachon
University,
Incheon
21936,
Republic
of
Korea.
3College
of
Pharmacy,
Gachon
University,
#191,
Hambakmoero,
Yeonsu-‐gu,
Incheon
21936,
Republic
of
Korea.
Sorbus
commixta
Hedl.
(Rosaceae),
also
known
as
Mountain
ash,
is
an
arbor
growing
in
the
mountainous
regions
and
distributed
throughout
Korea.
The
stem
bark
of
this
plant
has
been
used
in
Korean
traditional
medicine
for
the
treatment
of
cough,
asthma,
bronchial
disorders,
gastritis,
and
dropsy
[1,2].
Previous
studies
showed
that
S.
commixta
has
antioxidative,
anti-‐
atherogenic,
anti-‐inflammatory,
anti-‐sclerotic
and
vascular
relaxant
activities
[3].
Moreover,
flavonoids,
triterpenes,
and
lignans
were
isolated
from
this
source
[2-‐4].
In
the
course
of
our
continuing
search
for
bioactive
constituents
from
Korean
medicinal
plants,
we
isolated
poten-‐
tial
neuroprotective
compounds
from
the
stem
bark
of
S.
commixta;
Column
chromatographic
purification
of
the
CHCl3
fraction
of
S.
commixta
MeOH
extract
led
to
isolation
of
a
new
fatty
acid,
sorcomic
acid
(1),
along
with
nine
known
analogues
(2-‐10).
The
chemical
structure
of
the
new
compound
(1)
was
determined
through
spectroscopic
data
(NMR
and
MS)
and
specif-‐
ic
optical
rotation.
All
the
isolated
compounds
(1-‐10)
were
evaluated
for
their
neuroprotec-‐
tive
and
anti-‐inflammatory
activities.
Compounds
1,
5,
and
7
exhibited
potent
inductive
effect
of
NGF
secretion
from
C6
glioma
cells
(233.40
±
12.82%,
194.40
±
8.05%
and
185.74
±
10.25%,
respectively)
and
compound
10
reduced
NO
levels
with
an
IC50
value
of
71.25
μM
in
murine
microglia
BV2
cells
stimulated
by
LPS.
Also,
the
isolates
(1-‐10)
were
tested
for
their
cytotoxic
activity
against
A549,
SK‑OV‑3,
SK‑MEL‑2,
and
HCT15
cell
lines
in
vitro
using
the
SRB
bioassay.
Sorcomic
acid
(1)
Acknowledgements:
This
research
was
supported
by
the
Basic
Science
Research
Program
through
the
National
Research
Foundation
of
Korea
(NRF)
funded
by
the
Ministry
of
Education,
Science
and
Technol-‐
ogy
(2013R1A1A2A10005315).
Keywords: Sorbus commixta, Rosaceae, fatty acid, neuroprotection
References:
[1] Yang
G,
An
HJ.
β-‐sitosteryl-‐3-‐O-‐β-‐glucopyranoside
isolated
from
the
bark
of
Sorbus
com-‐
mixta
ameliorates
pro-‐inflammatory
mediators
in
RAW
264.7
macrophages.
Im-‐
munopharmacol
Immunotoxicol
2014;
36:
70-‐77
[2] Na
MK,
An
RB,
Min
BS,
Lee
SM,
Kim
YH,
Bae
KH.
Chemical
constituents
from
Sorbus
com-‐
mixta.
Nat
Prod
Sci
2002;
8:
62-‐65
[3] Bhatt
LR,
Bae
MS,
Kim
BM,
Oh
GS,
Chai
KY.
A
chalcone
glycoside
from
the
fruits
of
Sorbus
commixta
Hedl.
Molecules
2009;
14:
5323-‐5327
[4] Na
MK,
An
RB,
Lee
SM,
Min
BS,
Kim
YH,
Bae
KH,
Kang
SS.
Antioxidant
compounds
from
the
stem
bark
of
Sorbus
commixta.
Nat
Prod
Sci
2002;
8:
26-‐29
P513
Antiviral
effect
of
Epimedium
koreanum
extract
Won-‐Kyung
Cho1,
Jong-‐Soo
Lee2,
and
Jin
Yeul
Ma1
1
KM
Application
Center,
Korea
Institute
of
Oriental
Medicine,
70
Chemdan-‐ro,
Dong-‐gu,
Daegu,
701-‐300,
Daegu,
South
Korea,2Department
of
Veterinary
Medicine,
Chungnam
National
University,
99
Daehak-‐ro,
Yuseong-‐gu,
Daejeon,
305-‐764,
South
Korea
Epimedium
koreanum
Nakai
(EKN)
[Berberidaceae]
has
been
extensively
used
in
traditional
Korean
and
Chinese
medicine
to
treat
a
variety
of
diseases,
such
as
infertility,
impotence,
neu-‐
rasthenia,
cardiovascular
diseases,
amnesia,
lumbago,
arthritis,
various
immune-‐modulatory
problems,
aphrodisiac,
and
anti-‐rheumatic
[1-‐3].
Despite
of
EKN’s
immune
modulatory
poten-‐
tial
[4],
its
antiviral
properties
and
mode
of
action
have
not
been
completely
investigated.
In
the
present
study,
the
broad
spectrum
of
antiviral
efficacy
of
EKN
water
extract
was
evaluated
in
vitro,
and
moreover,
the
protective
effect
against
divergent
influenza
A
subtypes
was
de-‐
termined
using
BALB/c
mice.
EKN
at
100
ug/ml
markedly
reduced
the
replication
of
Influenza
A
Virus,
Vesicular
Stomatitis
Virus,
Herpes
Simplex
Virus
and
Newcastle
Disease
Virus
in
RAW264.7
and
HEK293T
cells.
EKN
at
100
ug/ml
induced
the
secretion
of
type
I
IFN
and
pro-‐
inflammatory
cytokines
and
the
subsequent
stimulation
of
antiviral
state
in
the
cells.
Among
various
components
in
the
extract,
quercetin
was
confirmed
to
have
striking
antiviral
proper-‐
ties.
The
oral
administration
of
EKN
at
20
mg
per
head
exhibited
preventive
effects
on
BALB/c
mice
infected
with
5
MLD50
influenza
A
subtypes
(H1N1,
H5N2,
H7N3
and
H9N2).
Therefore,
EKN
water
extract
plays
roles
as
immune-‐modulators
in
innate
immune
response,
and
may
be
potential
candidates
for
therapeutic
treatments
against
diverse
viruses
in
animal
and
humans.
Acknowledgements:
This
work
was
supported
by
a
grant
(Grant
No.
K12050)
awarded
to
the
Korean
Institute
of
Oriental
Medicine
by
the
Ministry
of
Education,
Science
and
Technology
(MEST),
Korea
and
the
Ministry
for
Food,
Agriculture,
Forestry
and
Fisheries,
Republic
of
Korea
(Grant
No.
112013033SB010)
Keywords: Epimedium koreanum Nakai, herbal medicine, antiviral effect, immune response
References:
[1] Chen
Y,
Zhao
YZ,
Jia
XB,
Hu
M.
Intestinal
absorption
mechanisms
of
prenylated
flavonoids
present
in
the
heat-‐processed
Epimedium
koreanum
Nakai.
Pharm
Res
2008;
25:
2190–
2199
[2] Ma
H,
He
X,
Yang
Y,
Li
M,
Hao
D.
The
genus
Epimedium:
An
ethno
pharmacological
and
photochemical
review.
J.
Ethnopharmacol.
2011;
134:
519–541
[3] Cho
WK,
Kim
H,
Choi
YJ,
Yim
NH,
Ma
JY.
Epimedium
koreanum
Nakai
water
extract
exhibits
antiviral
activity
against
porcine
epidermic
diarrhea
virus
in
vitro
and
in
vivo.
Evid
Based
Complement
Altern
Med
2012;
2012:
985151
[4] Ou
X,
Li
W.
Effect
on
enhancing
physical
strength
and
anti-‐stress
activity
of
flavonoids
from
the
Chinese
medicinal
plant
Epimedium
koreanum
Nakai.
Sci
Res
Essays
2010;
5:
883–886
P514
New
labdane
diterpenoids
from
Dysoxylum
hongkongense
Yao-‐Ching
Tsai1,
Tsong-‐Long
Hwang2,
Yuh-‐Chi
Kuo3,
Ching-‐Te
Chien4,
Ya-‐Ching
Shen1
1
School
of
Pharmacy,
National
Taiwan
University,
Taipei,
Taiwan,
2
Graduate
Institute
of
Natural
Prod-‐
ucts,
Taoyuan,
Taiwan,
3
Department
of
Life
Science,
Fu-‐Jen
University,
Taipei
Hsien,
Taiwan,
4
Division
of
Silviculture,
Taiwan
Forestry
Research
Institute,
Taipei,
Taiwan
The
leaves
and
bark
of
the
genus
Dysoxylum
were
used
as
a
remedy
by
the
indigenous
people
for
the
treatment
of
fever,
pain,
breeding,
malaria
and
convulsions.
Dysoxylum
hongkongense
(Tutch.)
Merr.
is
a
timber
tree,
which
grows
in
lowland
forest
of
southern
Taiwan
[1].
Our
pre-‐
liminary
biological
screening
revealed
that
the
ethanolic
extract
of
the
leaves
of
D.
hongkongense
showed
significant
inhibition
on
superoxide
anion
generation
(76%)
and
elas-‐
tase
release
(70%)
activities
at
10
µg/mL.
Because
no
prior
chemical
constituent
of
D.
hongkongense
was
reported,
the
research
of
the
anti-‐inflammatory
origin
was
initiated.
The
leaves
and
twigs
of
Taiwanese
D.
hongkongense
were
extracted
with
ethanol
thrice.
Fractiona-‐
tion
and
purification
of
diterpenoids
were
achieved
by
extensive
chromatography
including
LH-‐20,
Si
gel
column,
and
HPLC.
Eight
new
diterpenoids
designated
dysongensins
A–H
(1-‐8)
along
with
the
known
3-‐ketosclareolide
(9)
were
isolated
[2].
These
compounds
were
deter-‐
mined
by
spectroscopic
methods,
especially
2D
NMR.
The
structure
and
relative
configuration
of
dysongensin
A
(1)
was
supported
by
chemical
correlation
with
3-‐ketosclareolide
(9).
The
structure
of
dysongensin
C
was
confirmed
by
single
crystal
X-‐ray
crystallographic
analysis.
All
the
isolated
compounds
have
been
evaluated
for
their
in
vitro
inhibitory
activity
against
the
HSV-‐1
virus
[3],
and
have
been
tested
on
superoxide
anion
generation
and
elastase
release
by
human
neutrophils
in
response
to
FMLP/CB
[4].
Compounds
5
and
8
exhibited
mild
anti-‐
inflammatory
effects
(31.3
±
6.6%
and
25.3
±
4.0%)
on
elastase
release
and
superoxide
anion
generation,
respectively.
Sorafenib
is
used
as
a
standard
compound
(75.4
±
3.8%
and
77.2
±
4.2%).
Acknowledgements:
The
authors
thank
the
Ministry
of
Science
and
Technology,
Republic
of
China
(NSC104–2113–M–002–003)
for
financial
support.
Keywords: Dysoxylum hongkongense, labdanes,, 2D-‐NMR, anti-‐inflammatory activities
References
[1] Chang,
C.
E.,
Flora
of
Taiwan,
2nd
ed.,
1993;
3:
556-‐561
[2] Aranda,
G.;
Elkortbi,
M.
S.;
Lallemand,
J.
Y.;
Neuman,
A.;
Hammoumi,
A.;
Facon,
I.;
Azerad,
R.
Tetrahedron
1991;
47:
8339-‐8350
[3] Kuo,
Y.
C.;
Lin,
L.
C.;
Tsai,
W.
J.
C.;
Chou,
J.
S.
Kung,
H.
Y.;
Ho,
H.
Int
J
Antimicrob
Agents
2002;
46:
2854–2864
[4] Hwang,
T.
L.;
Yeh,
S.
H.,
Leu,
Y.
L.,
Chern,
C.
Y.,
Hsu,
H.
C.
Br
J
Pharmacol
2006;
148:
78-‐87
P515
Anti-‐inflammatory
diterpenoids
from
Callicarpa
randaiensis
Ho-‐Hsi
Cheng1,Yuan-‐Bin
Cheng2,
Tsong-‐Long
Hwang3,
Yao-‐Haur
Kuo4,
Ya-‐Ching
Shen
1
1
School
of
Pharmacy,
National
Taiwan
University,
Taipei
100,
Taiwan;
2
Graduate
Institute
of
Natural
Products,
College
of
Pharmacy,
Kaohsiung
Medical
University,
Kaohsiung
807,
Taiwan;
3
Graduate
Insti-‐
tute
of
Natural
Products,
College
of
Medicine,
Chang
Gung
University,
Taoyuan
333,
Taiwan;
4
Division
of
Herbal
Drugs
and
Natural
Products,
National
Research
Institute
of
Chinese
Medicine,
Taipei,
112,
Taiwan
Members
of
the
genus
Callicarpa
(beautyberries)
have
been
used
historically
in
Traditional
Chinese
Medicine
for
the
treatment
of
arthritis,
abdominal
pain,
bleeding,
common
cold,
and
cough
[1].
The
chemical
constituents
of
these
plants,
includes
diterpenoids,
lignans,
glyco-‐
sides,
and
flavonoids.
The
diterpenoids
of
Callicarpa
were
classified
into
abietane,
clerodane,
isopimarane,
labdane,
phyllocladane,
and
totaran
classes
[2].
C.
randaiensis
Hayata
(Verbena-‐
ceae)
is
an
endemic
shrub
that
grows
in
the
mountainous
area
of
central
Taiwan.
In
a
prelimi-‐
nary
biological
screening,
the
ethanolic
extract
of
the
leaves
from
C.
randaiensis
showed
sig-‐
nificant
inhibition
on
superoxide
anion
(95.5+
1.0
%)
and
elastase
release
(99.9
+
1.9
%)
in
human
neutrophils
at
a
concentration
of
10
µg/mL.
The
aim
of
the
work
was
to
investigate
the
origin
of
bioactivity,
including
isolation,
structural
elucidation
and
pharmacological
evalua-‐
tion
of
the
isolated
diterpenoids.
Leaves
and
twigs
of
C.
randaiensis
were
extracted
with
ethanol
and
nine
new
compounds,
callirandainins
A−I
(1-‐9)
were
isolated
in
addition
to
five
known
compounds,
3-‐oxo-‐17α,20α-‐
cleroda-‐13(14)-‐en-‐15,16-‐olide,
3α,4β-‐dihydroxy-‐17α,20α-‐cleroda-‐13(14)-‐en-‐15,16-‐olide,
3α-‐
hydroxy-‐17α,20α-‐cleroda-‐5(6),13(14)-‐dien-‐15,16-‐olide,
dehydroabietic
acid
and
β-‐amyrin.
Callirandainins
A−F
(1-‐6)
are
members
of
the
clerodane
diterpenoids,
the
callirandainins
G
(7)
and
H
(8)
possess
the
halimane
scaffold,
while
callirandainin
I
(9)
is
an
abietane-‐type
diterpe-‐
noid.
The
structures
of
the
new
compounds
were
established
on
the
basis
of
1D
and
2D
NMR
analysis
and
HRMS.
Anti-‐inflammatory
[3]
and
cytotoxic
activities
[4]
of
these
compounds
were
tested
and
evaluated.
In
the
anti-‐inflammatory
assays,
callirandainin
I
exhibited
potent
inhibition
of
both
superoxide
anion
(IC50
=
1.9
μM)
and
elastase
release
(IC50
=
2.1
μM).
LY294002
was
used
as
a
positive
control.
In
coclusion
nine
new
diterpenoids
callirandainins
A−I
(1-‐9)
were
isolated
from
C.
randaiensis
of
which
only
callirandainin
I
(9)
exhibited
potent
anti-‐inflammatory
activities.
Acknowledgements:
The
authors
thank
the
Ministry
of
Science
and
Technology,
Republic
of
China
(MOST
103-‐2320-‐B-‐002-‐010)
for
financial
support
Keywords: Callicarpa randaiensis, diterpenoids, anti-‐inflammatory activities
References:
[1] Committee
on
Chinese
Medicine
Pharmacy.
The
Illustration
of
Common
Medicinal
Plants
in
Taiwan
III,
Taipei,
2004,
pp.
40
[2] Tu
YH,
Sun
LN,
Guo
ML,
Chen
WS.
The
medicinal
uses
of
Callicarpa
L.
in
traditional
Chinese
medicine:
An
ethnopharmacological,
phytochemical
and
pharmacological
review.
J
Eth-‐
nopharmacol
2013;
146:
465‒481
[3] Yu
HP,
Hsieh
PW,
Chang
YJ,
Chung
PJ,
Kuo
LM,
Hwang
TL.
2-‐(2-‐Fluorobenzamido)benzoate
ethyl
ester
(EFB-‐1)
inhibits
superoxide
production
by
human
neutrophils
and
attenuates
hemorrhagic
shock-‐induced
organ
dysfunction
in
rats.
Free
Rad
Biol
Med
2011;
50:
1737−1748
[4] Arakawa
Y,
Saito
S,
Yamada
H,
Aiba
K.
Simultaneous
treatment
with
camptothecin
and
valproic
acid
suppresses
induction
of
Bcl-‐X(L)
and
promotes
apoptosis
of
MCF-‐7
breast
cancer
cells.
Apoptosis
2009;
14:
1076−1085
P516
Inhibitory
effects
of
andaliman
essential
oil
on
acid
production,
volatile
sulfur
compounds,
and
Actinomyces
viscosus
biofilms
Yanti,
Berti
Priska
Gea,
Bibiana
W.
Lay,
Marco
Tjakra,
Stevhen
Juniardi
Faculty
of
Biotechnology,
Atma
Jaya
Catholic
University,
Jalan
Jenderal
Sudirman
51,
Jakarta
12930,
Indonesia
Halitosis
(oral
malodour)
is
mainy
caused
by
the
excessive
production
of
acid,
oral
levels
of
volatile
sulfur
compounds
(VSCs),
and
accumulation
of
biofilm
plaque
by
oral
bacteria
in
the
mouth
cavity.
Andaliman
fruit
(Zanthoxylumacanthopodium)
is
an
endemic
spice
in
the
Ta-‐
panuli
region
(North
Sumatera
province)
that
has
been
reported
for
its
potential
antimicrobi-‐
al
and
anti-‐inflammatory
activities.
[1,2]
The
aim
of
this
research
was
to
determine
the
effect
of
andaliman
essential
oil
(20-‐100
µg/mL)
for
combating
halitosis
using
the
Actinomyces
vis-‐
cosus
biofilm
model
in
vitro
by
conducting
a
pH-‐stat
assay,
VSCs
quantification,
and
biofilm
quantification
assays.
Chromatographic
results
showed
that
andaliman
essential
oil
contained
76%
of
the
main
compound
of
carveol.
A.
viscosus
was
able
to
rapidly
produce
acid
in
20
minutes,
resulting
in
5.57
pH
from
pH
7.00,
and
andaliman
essential
oil
at
100
µg/mL
exerted
significant
inhibition
on
acid
production
from
A.
viscosus.
Andaliman
essential
oil
reduced
>50%
of
VCSs
produced
by
A.
viscosus
in
vitro
at
the
selected
dose.
Andaliman
essential
oil
showed
dual
antibiofilm
effects
on
preventing
A.
viscosus
biofilm
growth
and
killing
the
estab-‐
lished
biofilm.
At
the
lowest
dose
(20
µg/mL),
andaliman
essential
oil
significantly
inhibited
and
removed
>50%
of
A.
viscosus
biofilms
in
vitro.
In
conclusion,
andaliman
essential
oil
may
be
offered
as
a
natural
substance
for
halitosis
treatment.
Keywords: Andaliman fruit, essential oil, Actinomyces viscosus, biofilm, halitosis
References:
[1] Yanti,
Pramudito
TE,
Nuriasari
N,
Juliana
K.
Lemon
pepper
fruit
extract
(Zanthoxylum
ac-‐
anthopodium
DC.)
suppresses
the
expression
of
inflammatory
mediators
in
lipopolysac-‐
charide-‐induced
macrophages
in
vitro.
Am
J
Biochem
Biotechnol
2011;
7:
190-‐195
[2] Kristanty
RE,
Suriawati
J.
The
Indonesian
Zanthoxylum
acanthopodium
DC.:
chemical
and
biological
values.
Int
J
PharmTech
Res
2015;
8:
313-‐321
P517
Cucurbitane-‐type
triterpenes
and
glycoside
from
the
rattan
of
wild
Momordica
charantia
and
their
anti-‐inflammatory
and
cyto-‐
toxic
activities
Li-‐Jie
Zhang1,
Hung-‐Tse
Huang1,
Chia-‐Ching
Liaw2,
Shih-‐Yen
Huang3,
Zhi-‐Hu
Lin1,
Yao-‐Haur
Kuo1,4
1
National
Research
Institute
of
Chinese
Medicine,
Ministry
of
Health
and
Welfare,
No.
155-‐1,
Sec.
2,
Li-‐
nong
St.,
Taipei,
112,
Taiwan,
2
R&D
Department,
Starsci
Biotech
Co.
Ltd.,
2F,
No.
348,
Sec.
7,
Chengde
Rd.,
Taipei,
112,
Taiwan,
3
Endemic
Species
Research
Institute,
No.
1,
Ming-‐Shen
East
Rd.,
Nantou
County,
552,
Taiwan,
4
Ph.D.
Program
for
the
Clinical
Drug
Discovery
from
Botanical
Herbs,
College
of
Pharmacy,
Tai-‐
pei
Medical
University,
No.
250,
Wuxing
St.,
Taipei,
110,
Taiwan
Momordica
charantia
L.
(Cucurbitaceae),
which
is
a
slender-‐stemmed
tendril
climber
and
widely
cultivated
as
a
vegetable
crop,
is
commonly
called
bitter
melon,
goya
in
Japanese,
and
kugua
in
China
and
Taiwan.
The
fruit
of
the
plant
has
been
used
extensively
in
folk
medicine
as
a
remedy
for
diabetes
in
Asia
in
Asia,
India,
and
South
America
[1,
2].
Four
new
cucurbi-‐
tane-‐type
triterpenes,
(23R)-‐7β-‐hydroxy-‐3β-‐O-‐malonyl-‐23-‐methoxycucurbita-‐5,24-‐diene-‐19-‐
al
(1),
(23E)-‐7β,25-‐dihydroxy-‐3β-‐O-‐methylmalonylcucurbita-‐5,23-‐diene-‐19-‐al
(2),
(23E)-‐7β-‐
hydroxy-‐3β-‐O-‐methylmalonyl-‐25-‐methoxycucurbita-‐5,23-‐diene-‐19-‐al
(3),
(23E)-‐7β,25-‐
dihydroxy-‐3β-‐O-‐crotonylcucurbita-‐5,23-‐diene-‐19-‐al
(4),
and
one
new
glycoside
7β-‐hydroxy-‐
3β-‐O-‐malonyl-‐cucurbita-‐5,24-‐diene-‐19-‐a-‐23-‐O-‐β-‐D-‐glucopyranoside
(5),
were
isolated
from
the
rattans
of
wild
Momordica
charantia.
The
structures
of
these
compounds
were
established
via
spectroscopic
analyses,
including
1D
and
2D
NMR,
IR,
and
MS
techniques.
Pharmacological
studies
on
the
anti-‐inflammatory
effect
revealed
that
compounds
1−5
exhibited
moderate
activity
based
on
the
anti-‐NO
production
assay.
In
addition,
compounds
1−5
also
exhibited
moderate
cytotoxicity
against
MCF-‐7,
Hep-‐G2,
HEp-‐2,
and
WiDr
human
tumor
cell
lines
with
IC50
values
all
bellow
8
μg/ml.
Acknowledgements:
Y.H.
Kuo
gratefully
acknowledges
the
financial
support
of
this
work
from
the
Minis-‐
try
of
Science
and
Technology
(MOST
104-‐2320-‐B-‐077-‐006-‐MY3).
Keywords: Cucurbitane-‐type triterpenes, Momordica chaeantia, anti-‐inflammatory, cytotoxi-‐
city
References:
[1] Grover
JK,
Yadav
SP.
Pharmacological
actions
and
potential
uses
of
Momordica
charantia:
a
review.
J
Ethnopharmacol
2004;
83:
123−132
[2] Abascal,
K,
Yarnell
E.
Using
bitter
melon
to
treat
diabetes.
J
Altern
Complement
Med
2005;
1:
179–184
P518
Effect
of
elicitors
on
bioactive
compound
accumulation
in
shoot
culture
of
Dioscorea
membranacea
Yaowapha
Jirakiattikul1,
Panumart
Rithichai1,
Thipsukon
Boonyuen1,
Srisopa
Ruangnoo2,
Arunporn
Itharat2
1Department
of
Agricultural
Technology,
Faculty
of
Science
and
Technology,
Thammasat
University,
Rangsit
campus,
Pathumthani
12120,
Thailand,
2Department
of
Applied
Thai
Traditional
Medicine,
Fac-‐
ulty
of
Medicine,
Thammasat
University,
Rangsit
campus,
Pathumthani
12120,
Thailand
Dioscorealide
B,
an
important
bioactive
compound,
is
isolated
from
Dioscorea
membranacea
Pierre
ex
Prain
&
Burkill.
(Dioscoreaceae)
[1].
The
rhizomes
have
been
widely
used
for
the
treatment
of
dermopathy,
lymphopathy,
inflammation,
cancers,
venereal
diseases
and
leprosy
[2].
In
vitro
propagation
of
this
medicinal
plant
can
be
achieved
and
the
regenerated
shoots
are
able
to
produce
bioactive
compounds.
Jasmonic
acid
(JA)
and
salicylic
acid
(SA),
the
biotic
elicitors,
have
been
used
to
induce
secondary
metabolites
in
several
medicinal
plants
includ-‐
ing
Hypericum
perforatum
L.
[3],
Morinda
elliptica
[4]
and
Panax
ginseng
[5].
In
an
attempt
to
increase
bioactive
compound
productivity
in
shoot
culture
of
D.
membranacea,
the
effect
of
various
concentrations
of
JA
and SA
were
investigated.
Elicitation
treatments
were
studied
by
culturing
6
week-‐old
shoots
on
100,
250
and
500
µM
JA
and
50,
100
and
200
µM
SA
for
3
weeks
comparing
with
Murashige
and
Skoog
(MS)
medium
supplemented
with
8.87
µM
BA
(6-‐benzyladenine)
as
a
control
treatment.
Dioscorealide
B
content
of
0.59±0.08%
w/w
was
the
highest
at
100
µM
JA
which
was
1.51
times
greater
than
the
control
treatment
(0.39
±
0.06%
w/w).
All
concentrations
of
SA
had
adversely
effect on dioscorealide
B
contents
(0.15
±
0.06
-‐
0.23
±
0.06%
w/w).
There
was
no
significant
difference
among
treatments
on
total
phenolic
contents
of
47.44
±
9.25
−
53.54
±
3.85
mgGAE/g
dry
extract
and
EC50
of
DPPH
radi-‐
cal
scavenging
activity
of
51.45
±
3.32
−
61.35
±
7.99
µg/ml.
Acknowledgements:
This
work
was
supported
by
the
Higher
Education
Research
Promotion
and
National
Research
University
Project
of
Thailand,
The
office
of
the
Higher
Education
Commission
Keywords: Dioscorea membranacea, dioscorealide B, jasmonic acid, salicylic acid
References:
[1] Itharat
A,
Houghton
PJ,
Eno-‐Amooquaye
E,
Burke
PJ,
Sampson
JH,
Raman
A.
In
vitro
cyto-‐
toxic
activity
of
Thai
medicinal
plants
used
traditionally
to
treat
cancer.
J
Ethnopharmacol
2004;
90:
33−38
[2] Tewtrakul
S,
Itharat
A.
Anti-‐allergic
substances
from
the
rhizomes
of
Dioscorea
membra-‐
nacea.
Bioorg
Med
Chem
2006;
14:
8707−8711
[3] Walker TS,
Bais
HP,
Vivanco
JM.
Jasmonic
acid-‐induced
hypericin
production
in
cell
sus-‐
pension
cultures
of
Hypericum
perforatum
L.
(St.
John’s
wort).
Phytochemistry 2002; 60:
289−293
[4] Chong
TM,
Abdullah
MA,
Lai
OM,
Nor’Aini
FM,
Lajis
NH.
Effective
elicitation
factors
in
Morinda
elliptica
cell
suspension
culture.
Process
Biochem
2005;
40:
3397−3405
[5] Yu
KW,
Gao
W,
Hahn
EJ,
Paek
KY.
Jasmonic
acid
improves
ginsenoside
accumulation
in
adventitious
root
culture
of
Panax
ginseng
C.A.
Meyer.
Biochem
Eng
J
2002;
11:
211−215
P519
Phenolic
compounds
of
Salvia
hypargeia
Fich.
&
Mey.
Yavuz
Bülent
Köse1,
Fatih
Göger2,
Gökalp
İşcan2
1
Department
of
Pharmaceutical
Botany,
Anadolu
University,
Faculty
of
Pharmacy,
26470,
Eskişehir,
Tur-‐
key, 2 Department of Pharmacognosy, Anadolu University, Faculty of Pharmacy, 26470, Eskişehir, Turkey
The
genus
Salvia
L.
(Sage)
belongs
to
the
Lamiaceae
family
and
includes
around
1000
species
worldwide.
There
are
99
species
registered
in
the
Flora
of
Turkey
and
55
of
them
are
endemic
[1].
Salvia
species
are
important
export
products
for
Turkey
and
they
are
generally
consumed
as
medicinal
tea
in
folk
medicine
[2].
In
the
present
study
phenolic
compounds
profile
of
Salvia
hypargea
was
determined
by
LC/MSMS
system.
For
this
purpose,
air
dried
aerial
parts
of
plant
material
were
macerated
with
MeOH
for
24h
using
an
orbital
shaker.
After
extraction
process,
the
solvent
was
evapo-‐
rated
to
dryness
under
reduced
pressure
at
40°C.
For
the
determination
of
phenolic
com-‐
pounds,
a
Shimadzu
20A
HPLC
system
coupled
to
an
Applied
Biosystems
3200
Q-‐Trap
LC-‐
MS/MS
instrument
equipped
with
an
ESI
ion
source
was
used.
Separation
of
the
compounds
was
performed
on
a
ODS
150
x
4,6
mm,
i.d.,
3
µm
particle
size,
octadecyl
silica
gel
analytical
column
operating
at
40ºC
at
a
flow
rate
of
0.3
mL/min.
Caffeic
acid
(0.04
%),
luteolin
glucoside
(0.04
%),
apigenin
glucoside
(2.03
%)
,rosmarinic
acid
(2.20
%),
luteolin
(0.09
%),
apigenin
(0.08
%)
and
rosmarinic
acid
derivative,
quercetin
hexoside,
apigenin
glucuronide
were
determined
as
major
compounds
of
the
extract.
Keywords: Salvia hypargeia, LC-‐ MS/MS, Phenolic
References:
[1] Martin
E,
Altınordu
F,
Celep
F,
Kahraman
A,
Doğan
M.
Karyomorphological
studies
in
sev-‐
en
taxa
of
the
genus
Salvia
(Lamiaceae)
in
Turkey.
Caryologia
2015;
68:
13-‐18
[2] Baytop
T.
Therapy
with
Medicinal
Plants
in
Turkey.
Istanbul:
Istanbul
Univ.
Press;
1999.
P520
Medicinal
plants
traded
in
traditional
markets
of
southern
part
of
Guizhou,
Southwest
China
Liya
Hong1,2,
Yizhou
Wang1,
Jianqin
Li1,3,
Qiyi
Lei1,4,
Jiangju
Zhou4,
Liang
Qin1,
Chunlin
Long1,5*
1
College
of
Life
and
Environmental
Sciences,
Minzu
University
of
China,
Beijing
100081,
China,
2School
of
Environmental
and
Biological
Sciences,
Rutgers,
The
State
University
of
New
Jersey,
New
Brunswick
08901,
USA,
3
Faculty
of
Forestry,
Southwest
Forestry
University,
Kunming
650224,
China,
4
School
of
En-‐
vironment
&
Life
Science,
Kaili
University,
Kaili
556011,
China,
5
Kunming
Institute
of
Botany,
Chinese
Academy
of
Sciences,
Kunming
650201,
China
Traditional
herbal
markets
have
existed
in
China
for
many
centuries,
especially
in
the
rural
areas
[1].
Ethnobotanical
market
survey
is
often
adopted
for
documenting
herbal
plants
and
associated
traditional
knowledge.
Over
3000
species
of
higher
plants
occur
in
southern
part
of
Guizhou,
southwest
China.
This
area
is
well-‐known
for
its
cultural
diversity.
Miao,
Dong,
Buyi,
Yao,
Shui,
Maonan
and
other
ethnic
groups
live
in
this
region.
Almost
all
ethnic
people
in
the
region
depend
on
local
medicinal
markets
for
medicinal
treatment
and
family
healthcare.
However,
information
on
the
market
traded
medicinal
plants
is
not
well
documented
from
traditional
medicinal
markets
in
southern
Guizhou.
The
study
aimed
to
document
medicinal
plants
traded
by
local
people
in
the
periodic
markets.
A
total
of
387
medicinal
plant
species,
belong
to
309
genera
and
123
families,
were
investigated
and
collected
together
with
their
medicinal
uses
by
the
local
people.
The
plants
were
used
to
treat
256
human
diseases.
Of
the
387
species,
234
(60.47%)
were
obtained
from
the
wild
whereas
71
(18.35%)
were
from
home
gardens,
and
82
(21.19%)
species
were
from
both
home
gardens
and
wild
habitats.
Herbaceous
species
(247)
were
the
most
utilized
plants,
accounting
for
63.82%
of
the
species,
followed
by
shrubs
(59,
15.25%).
Fabaceae
came
out
as
a
leading
family
with
25
medicinal
species
(6.46%)
while
Asteraceae
(5.94%)
followed
with
23
species.
The
most
frequently
used
plant
parts
were
the
whole
plants
(148),
followed
by
roots
(84).
Decoction
was
a
widely
used
method
to
prepare
traditional
herbal
medicines.
Our
research
results
indicated
that
the
study
area
might
be
probably
one
of
the
richest
diversity
centers
in
both
species
and
cultural
level
of
the
country’s
ethnomedicines,
to
compare
with
other
parts
of
China
[2-‐4].
The
medici-‐
nal
plants
which
spontaneously
sold
in
the
open
air
markets
are
collected
by
the
local
people
from
wild
and
their
living
environment,
even
the
toxic
plants.
There
is
at
risk
of
ingesting
tox-‐
ic
plants
sold
in
popular
markets,
because
there
is
no
hard
distinction
between
medicinal
and
toxic
plants.
The
local
government
should
regulate
the
traditional
markets
for
misusing
of
the
herbal
medicine
and
protecting
the
local
biodiversity
and
associated
traditional
knowledge.
Acknowledgements:
The
local
informants
and
healers
are
fully
acknowledged
for
their
participation
and
sharing
their
valuable
knowledge
on
the
use
of
medicinal
plants
with
us.
This
work
was
financially
sup-‐
ported
by
the
National
Natural
Science
Foundation
of
China
(31161140345),
the
Ministry
of
Science
and
Technology
of
China
(2012FY110300),
Minzu
University
of
China
(YLDX01013,
2015MDTD16C),
and
the
Ministry
of
Education
of
China
(B8044).
Keywords:
Medicinal
plants,
traditional
knowledge,
periodic
market,
southern
part
of
Gui-‐
zhou,
medical
ethnobotany
References:
[1] Lee
S,
Xiao
C,
Pei
S.
Ethnobotanical
survey
of
medicinal
plants
at
periodic
markets
of
Honghe
Prefecture
in
Yunnan
Province,
SW
China.
J
Ethnopharmacol
2008;
117:
362-‐377
[2] Long
CL,
Li
SM,
Long
B,
Shi
YN,
Liu
BX.
Medicinal
plants
used
by
the
Yi
ethnic
group:
a
case
study
in
central
Yunnan.
J
Ethnobiol
Ethnomed
2009;
5:
13
[3] Liu
YC,
Dao
ZL,
Yang
CY,
Liu
YT,
Long
CL.
Medicinal
plants
used
by
the
Tibetans
in
Shangri-‐
la,
Yunnan,
China.
J
Ethnobiol
Ethnomed
2009;
5:
15
[4] Hong
LY,
Guo
ZY,
Huang
KH,
Wei
SJ,
Liu
B,
Meng
SW,
Long
CL.
Ethnobotanical
study
on
me-‐
dicinal
plants
used
by
Maonan
people
in
China.
J
Ethnobiol
Ethnomed
2015;
11:32
P521
Comparing
the
leaf
secretory
apparatus
of
Hibiscus
sabdariffa
and
Hibiscus
surattensis
K.
Raghu,
Y.
Naidoo
School of Life Sciences, University of KwaZulu-‐Natal, Private Bag X54001, Durban, 4000, South Africa
Floral
biodiversity
throughout
the
world
is
declining
rapidly
owing
to
detrimental
human
ac-‐
tivities.
There
is
an
urgency
to
screen
and
evaluate
plants
with
reported
ethnobotanical
value.
The
medicinal
activities
of
plants
can
be
attributed
to
special
phytochemicals
produced
through
specific
secondary
metabolic
pathways
within
plant
secretory
structures.
Under-‐
standing
the
mechanisms
of
production
and
secretion
in
glandular
and
anatomically
complex
secretory
structures
may
lead
to
advances
in
optimizing
the
yield
and
activities
of
compounds
with
pharmacological
benefits.
This
study
was
undertaken
to
examine
and
evaluate
structures
on
the
leaves
of
two
well-‐known
traditionally-‐used
medicinal
plant
species,
Hibiscus
suratten-‐
sis
and
Hibiscus
sabdariffa.
This
was
achieved
using
a
range
of
light
and
electron
microscopy
techniques.
Stalk-‐bearing
capitate
glandular
trichomes
were
identified
as
the
main
external
secretory
appendages
found
on
foliar
surfaces
of
both
Hibiscus
species.
Histochemical
staining
revealed
the
presence
of
acidic
polysaccharides,
phenolics
and
alkaloids
in
trichome
head
cells
of
H.
surattensis
and
H.
sabdariffa.
Although
similar
in
morphology
and
chemical
compo-‐
sition,
the
ultrastructure
of
the
capitate
glands
differed
significantly
between
the
species.
Those
of
H.
surattensis
showed
high
vacuolation
of
trichome
head
cells
with
dense
porous
ag-‐
gregates,
thought
to
be
phenolics,
occurring
within.
Capitate
trichomes
of
H.
sabdariffa
ap-‐
peared
to
compartmentalise
electron-‐dense
amorphous
deposits
in
the
dorsal
region
of
the
head.
The
trichomes
of
both
species
demonstrated
the
presence
of
a
subcuticular
space
on
the
ventral
surface
of
the
trichome,
into
which
amorphous
substances
were
secreted.
Due
to
the
high
levels
of
vesiculation
within
head
cells
of
both
the
species
studied,
secretion
into
the
sub-‐
cuticular
space
is
assumed
to
be
granulocrine
in
nature.
In
addition
to
external
foliar
glands,
internal
mucilage
repositories
were
also
identified
in
H.
surattensis
and
H.
sabdariffa.
Mucilage
idioblasts
stained
intensely
for
acidic
polysaccharides
and
acidic
lipids
and
were
shown
to
develop
from
the
deposition
of
mucilage
into
the
periplasmic
space
between
the
protoplast
and
cell
wall.
Mucilage
conducting
ducts,
located
among
the
vascular
tissues
appeared
to
be
formed
through
schizogenous
processes.
The
surrounding
epithelium
demonstrated
subcellu-‐
lar
processes
indicative
of
mucilage
secretion
into
the
duct.
The
findings
here
form
the
basis
of
further
exploration
into
the
secretory
mechanisms
of
both
capitate
trichomes
and
mucilage
repositories
in
Hibiscus.
The
prominence
of
phenolic
and
alkaloid
compound
classes
within
foliar
trichomes
of
both
Hibiscus
species
may
be
linked
to
their
antihypertensive,
antilipidem-‐
ic
and
anticancer
properties.
Further
investigation
into
the
biological
activities
of
the
extracts
and
their
derived
compounds
is
presently
underway.
Acknowledgements: National Research Foundation (South Africa) for funding this study.
Keywords: Hibiscus, micromorphology, capitate trichome, mucilage, electron-‐dense deposits
References:
[1] Herrera-‐Arellano
A,
Flores-‐Romero
S,
Chavez-‐Soto
MA,
Tortoriello
J.
Effectiveness
and
tol-‐
erability
of
a
standardized
extract
from
Hibiscus
sabdariffa
in
patients
with
mild
to
mod-‐
erate
hypertension:
a
controlled
and
randomized
clinical
trial.
Phytomedicine
2004;
11:
375-‐382
[2] Valkama
E,
Salminen
J,
Koricheva
J,
Pihlaja
J.
Comparative
analysis
of
leaf
trichome
struc-‐
ture
and
composition
of
epicuticular
flavonoids
in
Finnish
birch
species.
Ann
Bot
2003;
91:
643-‐
655
P522
Soluble
epoxide
hydrolase
inhibitory
activity
of
anthraquinone
components
from
Aloe
Ya
Nan
Sun,
Ah
Reum
Jo,
Jang
Hoon
Kim,
Jong
Seong
Kang,
Young
Ho
Kim
College
of
Pharmacy,
Chungnam
National
University,
Daejeon
305-‐764,
Korea
Aloe
is
a
short-‐stemmed
succulent
herb
widely
used
in
traditional
medicine
to
treat
various
diseases
and
as
raw
material
in
cosmetics
and
heath
foods
[1,
2].
In
this
study,
we
isolated
and
identified
two
new
anthraquinone
derivatives,
aloinoside
C
(6)
and
aloinoside
D
(7),
together
with
six
known
compounds
from
an
aqueous
dissolved
Aloe
exudate.
Their
structures
were
identified
by
spectroscopic
analysis
[2-‐5].
The
inhibitory
effects
of
the
isolated
compounds
on
soluble
epoxide
hydrolase
(sEH)
were
evaluated
[3,
4].
Compounds
1–8
inhibited
sEH
activity
potently,
with
IC50
values
ranging
from
4.1
±
0.6
to
41.1
±
4.2
μM.
A
kinetic
analysis
of
com-‐
pounds
1–8
revealed
that
the
inhibitory
actions
of
compounds
1,
6
and
8
were
non-‐
competitive,
whereas
those
of
compounds
2–5
and
7
were
the
mixed-‐type.
Molecular
docking
increases
our
understanding
of
receptor-‐ligand
binding
of
all
compounds.
The
Autodock
scores
for
1–8
were
-‐7.26,
-‐6.92,
-‐7.81,
-‐7.38,
-‐7.38,
-‐8.51,
-‐7.71,
and
-‐8.59
kcal/mol,
respec-‐
tively.
These
results
demonstrate
that
compounds
1–8
from
Aloe
are
potential
sEH
inhibitors.
Keywords:
Aloe,
anthraquinone
derivatives,
soluble
epoxide
hydrolase
(sEH),
molecular
sim-‐
ulation
References:
[1] Zhong
JS,
Huang
YY,
Ding
WJ,
Wu
XF,
Wan
JZ,
Luo
HB.
Chemical
constituents
of
Aloe
barba-‐
densis
Miller
and
their
inhibitory
effects
on
phosphodiesterase-‐4D.
Fitoterapia
2013;
91:
159-‐165
[2] Abdissa
N,
Induli
M,
Fitzpatrick
P,
Alao
JP,
Sunnerhagen
P,
Landberg
G,
Yenesew
A,
Er-‐
délyi
M.
Cytotoxic
quinones
from
the
roots
of
Aloe
dawei.
Molecules
2014;
19:
3264-‐3273
[3] Bbosa
GS,
Kyegombe
DB,
Lubega
A,
Musisi
N,
Ogwal-‐Okeng
J,
Odyek
O.
Antiplasmodium
falciparum
activity
of
Aloe
dawei
and
Justicia
betonica.
Afr
J
Pharm
Pharmacol
2013;
7:
2258-‐2263
[4] Kim
JH,
Ryu
YB,
Lee
WS,
Kim
YH.
Neuraminidase
inhibitory
activities
of
quaternary
iso-‐
quinoline
alkaloids
from
Corydalis
turtschaninovii
rhizome.
Bioorg
Med
Chem
2014;
22:
6047-‐6052
[5] Zhong
JS,
Huang
YY,
Zhang
TH,
Liu
YP,
Ding
WJ,
Wu
XF,
Xie
ZY,
Luo
HB,
Wan
JZ.
Natural
phosphodiesterase-‐4
inhibitors
from
the
leaf
skin
of
Aloe
barbadensis
Miller.
Fitoterapia
2015;
100,
68-‐74
P523
Lignans
with
inhibitory
activity
against
PCSK9
mRNA
expression
from
the
fruits
of
Schisandra
chinensis
Young-‐Mi Kim, Young Hee Choi, Young-‐Won Chin
College
of
Pharmacy
and
Intergrated
Research
Institute
of
Drug
Development,
Dongguk
University-‐Seoul,
32,
Dongguk-‐lo,
Goyang-‐si,
Gyeonggi-‐do
410-‐820,
Korea
Bioactivity-‐guided
fractionation
on
the
fruits
of
Schisandra
chinensis
(Turcz.)
Baill.
[Schisan-‐
draceae]
,
using
PCSK9
mRNA
expression
screening
assay,
led
to
the
isolation
of
two
new
lignans,
tigloylschinlignans
D
(1)
and
1-‐(3-‐hydroxy-‐4,5-‐dimethoxyphenyl)-‐4-‐(3ˈ-‐hydroxy-‐4ˈ-‐
methoxyphenyl)-‐2,3-‐dimethylbutane-‐1,4-‐diol
(2)
along
with
28
known
compounds.
All
the
structures
were
established
by
NMR
spectroscopic
data,
CD
and
MS
analysis.
All
the
isolates
were
tested
for
their
inhibitory
activities
on
mRNA
expression
of
proprotein
convertase
sub-‐
tilisin-‐kexin
type
9
(PCSK9),
which
is
known
to
modulate
low-‐density
liporprotein
receptor
[1-‐3].
Several
of
the
isolated
compounds
potently
inhibited
PCSK9
mRNA
expressions
with
IC50
values
between
0.36-‐3.85
µM,
including
the
new
lignan
2
with
IC50
3.15
µM.
Acknowledgements:
This
study
was
supported
by
the
grants
from
the
National
Research
Foundation
of
Korea
(NRF)
grant
funded
by
the
Korea
government
(MSIP)
(NRF-‐2015R1A2A2A01006736)
and
from
the
GRRC
program
of
Gyeonggi
province
[GRRC
DONGGUK2016–B01
and
B03]
Keywords: Schisandra chinensis, PCSK9, lignan, hypercholestrolemia
References:
[1] Soutar
AK,
Naoumova
RP.
Mechanisms
of
disease:
genetic
causes
of
familial
hypercholes-‐
terolemia.
Nat
Clin
Prac
2007;
4:
214−225
[2] Dadu
RT,
Ballantyne
CM.
Lipid
lowering
with
PCSK9
inhibitors.
Nat
Rev
Cardiol
2014;
11:
563−575
[3] Norata
GD,
Tibolla
G,
Catapano
AL.
PCSK9
inhibition
for
the
treatment
of
hypercholester-‐
olemia:
Promises
and
emerging
challenges.
Vascular
Pharmcol
2014;
62:
103−111
P524
Chemical
constituents
and
bioactivities
of
the
twigs
of
Severinia
buxifolia
Yuan-‐Bin
Cheng1,2,
Pei-‐Shian
Li1,
Yang-‐Chang
Wu3,4,5,
Fang-‐Rong
Chang1
1
Graduate
Institute
of
Natural
Products,
College
of
Pharmacy,
2
Research
Center
for
Natural
products
&
Drug
Development,
Kaohsiung
Medical
University,
80708
Kaohsiung,
Taiwan;
3
School
of
Pharmacy,
Col-‐
lege
of
Pharmacy,
China
Medical
University,
40402
Taichung,
Taiwan;
4
Chinese
Medicine
Research
and
Development
Center;
5
Center
for
Molecular
Medicine,
China
Medical
University
Hospital,
Taichung
40402,
Taiwan
Severinia
buxifolia
(Rutaceae)
is
an
evergreen
coastal
bush
distributing
over
temperate
and
sub-‐tropical
regions.
This
plant
is
used
as
a
folk
medicine
for
the
treatment
of
malaria,
rheu-‐
matism,
anti-‐nociceptive
and
phlegm
in
Southeast
Asia
[1].
In
previous
pharmacological
stud-‐
ies,
S.
buxifolia
was
proved
to
have
anti-‐inflammatory,
antiviral,
cytotoxic
activities
[2‒4].
In
our
bioactive
study,
the
methanolic
extract
of
the
twigs
of
S.
buxifolia
showed
promising
anti-‐
hepatitis
C
virus
and
cytotoxic
activities.
Thus,
this
plant
material
was
chosen
for
further
phy-‐
tochemical
investigation.
The
twigs
of
S.
buxifolia
(2.1
kg)
were
extracted
by
methanol
and
separated
by
a
series
of
column
chromatography.
As
a
result,
twenty
known
and
five
new
compounds
named
buxifolins
A‒E
(1‒5)
were
isolated.
The
structures
of
all
isolates
can
be
classified
into
acridone
alkaloids,
triterpenoids,
flavonids,
coumarins,
benzoates,
and
al-‐
dimines.
Cytotoxic
activities
of
the
isolates
were
tested
and
evaluated
by
the
MTT
assay.
Among
all
tested
compounds,
atalaphyllidine
(6)
demonstrated
potent
cytotoxicity
against
HepG2
cancer
cell
line
(IC50
=
2.25
μg/mL).
Acknowledgements:
The
authors
thank
the
grants
from
ministry
of
science
and
technology
of
Taiwan
(MOST103-‐2628-‐B-‐037-‐001-‐MY3
award
to
Y.-‐B
C).
The
authors
also
thank
the
support
from
Kaohsiung
medical
university
(Aim
for
the
Top
Universities
Grant,
grant
No.
KMU-‐TP104H02).
Keywords: Severinia buxifolia, acridone, cytotoxicity
References:
[1] Liu
TS,
Lai
MC.
Flora
of
Taiwan,
1977;
2:
525
[2] Wu
TS,
Leu
YL.
Tetranortriterpenoid
insect
antifeedants
from
Severinia
buxifolia.
Phyto-‐
chemistry
1997;
45:
1393–1398
[3] Wu
TS,
Leu
YL,
Chan
YY.
A
new
quinolin-‐2,4-‐dione
and
other
constituents
from
Severinia
buxifolia.
Phytochemistry
1998;
49:
1467–1470
[4] Wu
TS,
Chen
CM,
Lin
FW.
Constituents
of
the
root
bark
of
Severinia
buxifolia
collected
in
Hainan.
J
Nat
Prod
2001;
64:
1040–1043
P525
Study
on
the
anti-‐melanoma
constituents
from
Pinus
taiwanensis
Hayata
Yu-‐Chen
Kao,
Jen-‐Der
Wu,
Ching-‐Kuo
Lee
School of Pharmacy, College of Pharmacy, 250 Wuxing Street, Taipei 110, Taipei City, Taiwan.
The
Pinus
taiwanensis
Hayata
(Pinaceae)
is
an
endemic
plant
of
Tawian,
known
as
Taiwan
red
pine
which
is
grown
naturally
and
widespread
at
an
elevation
of
750-‐3500
m
in
the
Central
Mountain
of
Taiwan
[1].
Previous
reports
of
Pinus
species
have
revealed
antimicrobial
[2],
antioxidant
[2],
and
anticancer
[3]
activities,
however,
there
are
few
studies
on
the
constitu-‐
ents
of
P.
taiwanensis.
The
aim
of
this
study
is
to
isolate
chemical
constituents
and
evaluate
the
anti-‐melanoma
effect
of
P.
taiwanensis.
In
vitro
treatment
of
the
B16-‐F10
cell
with
the
eth-‐
anolic
extract
(25-‐100
μg/ml)
reduced
cell
viability
in
a
dose-‐dependent
manner.
The
extract
of
P.
taiwanensis
was
suspended
in
H2O
and
partitioned
successively
with
ethyl
acetate
(EtOAc)
and
n-‐butanol
(BuOH).
Each
extract
was
evaporated
to
dryness
under
reduced
pres-‐
sure
to
yield
ethyl
acetate
and
n-‐butanol
fractions,
respectively.
The
ethyl
acetate
layer
re-‐
vealed
significant
cytotoxic
effect
(inhibitory
rate:
94.58
± 0.22
%,
100
μg/ml),
and
which
was
further
subjected
to
silica
gel
open
column
and
high
performance
liquid
chromatography
(HPLC).
Bioactivity-‐guided
fractionation
led
to
the
isolation
of
a
series
of
compounds.
Their
structures
were
established
on
the
basis
of
extensive
spectroscopic
(IR,
MS,
2D
NMR)
data
analysis,
and
by
the
comparison
with
spectroscopic
data
reported
in
the
literatures.
The
new
diterpene
9α,13α-‐dihydroxyabiet-‐8(14)-‐enoic
acid
(4)
exhibits
no
anti-‐melanoma
activity
at
concentrations
less
than
50
μg/ml.
Acknowledgements:
The
authors
are
grateful
to
Ministry
of
Science
and
Technology,R.O.C.
for
financial
assistance
(NSC
101-‐2320-‐B-‐038-‐013-‐MY3,
MOST104-‐2320-‐B-‐038-‐020-‐MY3)
Keywords: Pinus taiwanensis Hayata, pinaceae, B16-‐F10 melanoma cells, HPLC
References:
[1] Lu
JJ,
Lin
KC,
Cheng
YS.
Terpenoids
From
Oleoresin
of
Pinus
taiwanensis.
Phytochemistry
1975;
14:
1375−1377
[2] Zeynep
U.
Chemical
composition,
antimicrobial,
insecticidal,
phytotoxic
and
antioxidant
activities
of
Mediterranean
Pinus
brutia
and
Pinus
pinea
resin
essential
oils.
Chinese
J
Nat
Med
2014;
12:
901−910
[3] Duan
JA.
Diterpenoids
from
Pinus
massoniana
resin
and
their
cytotoxicity
against
A431
and
A549
cells.
Phytochemistry
2010;
71:
1528−1533
P526
Anti-‐inflammatory
activity
of
resveratrol
metabolites
Yulia
Radko1,
Steen
B.
Pedersen2,
Lars
P.
Christensen1
1
Department
of
Chemical
Engineering,
Biotechnology
and
Environmental
Technology,
Faculty
of
Engi-‐
neering,
University
of
Southern
Denmark,
Campusvej
55,
5230
Odense
M,
Denmark;
2
Department
of
Clin-‐
ical
Medicine,
Aarhus
University,
Tage-‐Hansens
Gade
2,
8000
Aarhus
C,
Denmark
Resveratrol
(3,
5,
4´-‐trihydroxy-‐trans-‐stilbene)
is
a
naturally
occurring
polyphenol.
It
has
been
shown
to
possess
a
set
of
interesting
pharmacological
activities,
including
influence
on
insulin
secretion
[1].
The
main
aim
of
this
study
was
to
explore
the
possibility
of
application
of
resveratrol
metabolites
in
the
treatment
of
inflammatory-‐related
diseases.
Anti-‐inflammatory
activity
of
resveratrol
metabolites
isolated
from
human
urine
[2]
was
investigated.
We
exam-‐
ined
the
effect
of
resveratrol
metabolites
on
lipopolysaccharide
(LPS)-‐stimulated
THP-‐1
cells
(human
monocytic
cell
line)
by
measuring
suppression
of
interleukin
6
(IL-‐6)
[3].
It
can
be
seen
from
the
experimental
data
(Figure
1),
that
resveratrol
metabolites
inhibit
production
of
IL-‐6,
but
are
less
active
than
the
parent
compound.
Resveratrol
sulfate
conjugates
seem
to
exhibit
more
effect
on
IL-‐6
than
the
resveratrol
glucuronide
conjugates.
Moreover,
the
bioac-‐
tivity
of
trans-‐resveratrol-‐3,
4’-‐O-‐disulfate
is
comparable
to
resveratrol.
Dihydroresveratrol-‐3-‐
O-‐β-‐D-‐glucuronide
not
previously
investigated
for
its
anti-‐inflammatory
activity,
did
not
show
suppression
of
IL-‐6
in
LPS-‐stimulated
THP-‐1
cells.
From
the
results
of
the
present
study
it
appears
that
resveratrol
metabolites
inhibit
production
of
IL-‐6,
although
to
a
lower
extent
than
the
parent
compound;
hence,
it
is
assumed
that
resveratrol
metabolites
can
also
contrib-‐
ute
to
the
anti-‐inflammatory
effect
after
oral
intake
of
resveratrol.
Figure1.
Effect
of
resveratrol
and
resveratrol
metabolites
on
production
of
IL-‐6.
Metabolites
were
tested
in
the
following
concentrations:
A:
30
µg/ml,
B:
10
µg/ml
and
C:
5
µg/ml.
Concen-‐
tration
of
resveratrol
(RSV)
was
108
µg/ml.
Note
that
the
bars
on
the
graph
are
extended
to
negative
values
to
facilitate
visibility.
Acknowledgements:
This
work
was
supported
by
the
Danish
Council
for
Strategic
Research
(Grant
10-‐093499)
Keywords: Resveratrol, resveratrol metabolites, anti-‐inflammatory activity
References:
[1] Nøhr
MK,
Dudele
A,
Poulsen
MM,
Ebbesen
LH,
Radko
Y,
Christensen
LP,
Jessen
N,
Richel-‐
sen
B,
Lund
A,
and
Pedersen
SB.
LPS-‐enhanced
glucose-‐stimulated
insulin
secretion
is
normalized
by
resveratrol.
PLoS
ONE
2016;
11:
e0146840
[2] Radko
Y,
Christensen
KB,
Christensen
LP.
Semi-‐preparative
isolation
of
dihydroresveratrol-‐
3-‐O-‐β-‐D-‐glucuronide
and
four
resveratrol
conjugates
from
human
urine
after
oral
intake
of
a
resveratrol-‐containing
dietary
supplement.
J
Chromatogr
B
2013;
930:
54–61
[3] Radko
Y.
Anti-‐inflammatory
studies
on
resveratrol,
its
conjugates,
and
selected
plant
ex-‐
tracts.
PhD
thesis,
University
of
Southern
Denmark,
2016
P527
Antimutagenic
potentials
of
flavonoids
from
Achillea
millefolium
L.
subsp.
millefolium
Handan
G.
Sevindik1,
Zühal
Güvenalp1,
Mehmet
Karadayı2,
Medine
Güllüce2,
L.
Ömür
Demi-‐
rezer3
1
Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Atatürk
University,
25240
Erzurum,
Turkey,
2
De-‐
partment
of
Biology,
Faculty
of
Science,
Atatürk
University,
25240
Erzurum,
Turkey,
3
Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Hacettepe
University,
06100
Ankara,
Turkey
Current
researches
have
showed
that
cancer
is
one
of
the
most
widespread
diseases
in
the
world
and
there
is
a
strong
connection
between
its
formation
and
mutagenesis.
This
has
at-‐
tracted
attention
toward
mutation-‐focused
researches
to
get
better
understanding,
and
find-‐
ing
new
molecules
with
antimutagenic
potential
is
a
key
concept
for
prevention
of
the
haz-‐
ardous
effects
of
mutagens
[1].
The
genus
Achillea
L.
(Asteraceae)
comprises
85
species
worldwide
and
50
species
in
the
flora
of
Turkey
with
more
than
50%
endemism
[2].
In
the
present
study
we
investigated
the
mutagenic
and
antimutagenic
potentials
of
flavonoids
iso-‐
lated
from
Achillea
millefolium
L.
subsp.
millefolium
by
using
Ames/Salmonella
and
E.
coli
WP2
bacterial
test
systems
[3].
Four
active
compounds
were
isolated
from
the
aerial
parts
of
the
plant
and
identified
as
luteolin,
apigenin
7-‐O-‐β-‐D-‐glucoside,
luteolin
7-‐O-‐β-‐D-‐glucoside
and
6-‐OH-‐luteolin
7-‐O-‐β-‐D-‐glucoside
by
using
a
combination
of
extensive
spectroscopic
anal-‐
ysis
and
chemical
methods.
All
of
these
compounds
showed
significant
antigenotoxic
activity
against
the
positive
mutagens
MNNG
and
9-‐AA.
The
inhibition
rates
ranged
from
26.7%
(lute-‐
olin
7-‐O-‐β-‐D-‐glucoside-‐3
µM/plate)
to
39.3%
(luteolin-‐5
µM/plate)
for
MNNG
induced
muta-‐
genesis
in
E.
coli
WP2uvrA,
and
from
27.4%
(apigenin
7-‐O-‐β-‐D-‐glucoside-‐4
µM/plate)
to
34.2%
(apigenin
7-‐O-‐β-‐D-‐glucoside-‐5
µM/plate)
for
9-‐AA
induced
mutagenesis
in
S.
typhi-‐
murium
TA1537.
25-‐40%
inhibition
was
defined
as
moderate
antimutagenicity;
40%
or
more
inhibition
as
strong
antimutagenicity;
and
25%
or
less
inhibition
as
no
antimutagenicity
[4].
Keywords:
Achillea
millefolium,
apigenin,
luteolin,
antimutagenicity,
Ames-‐Salmonella,
E.
coli
WP2
References:
[1] Bhattacharya
S.
Natural
Antimutagens:
A
Review.
Res
J
Med
Plant
2011;
5:
116–126.
[2] Huber-‐Morath
A.
Flora
of
Turkey
and
East
Aegean
Islands.
In:
Davis
PH,
editor.
University
Press:
Edinburgh,
1978,
Volume
6.
[3] Mortelmans
K,
Zeiger
E.
The
Ames
Salmonella/microsome
mutagenicity
assay.
Mutat
Res
2000;
55:
29–60.
[4] Güllüce
M,
Agar
G,
Baris
O,
Karadayi
M,
Orhan
F,
Şahin
F.
Mutagenic
and
antimutagenic
effects
of
some
Astragalus
species
grown
in
the
Eastern
Anatolia
Region
of
Turkey.
Phy-‐
tother
Res
2010;
24:
1014–1018.
P528
Active
compounds
from
Achillea
biebersteinii
and
determination
of
their
antigenotoxic
potentials
Handan
G.
Sevindik1,
Zühal
Güvenalp1,
Mehmet
Karadayı2,
Medine
Güllüce2,
L.
Ömür
Demi-‐
rezer3
1Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Atatürk
University,
25240
Erzurum,
Turkey,
De-‐
partment
of
Biology,
Faculty
of
Science,
Atatürk
University,
25240
Erzurum,
Turkey,
3
Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Hacettepe
University,
06100
Ankara,
Turkey
In
this
context,
finding
new
sources
of
antigenotoxic/antimutagenic
compounds
that
enable
development
of
protective
or
therapeutic
drugs
remains
as
an
attractive
research
subject.
Therefore,
the
aim
of
the
present
study
was
to
determine
antimutagenic
constituents
of
Achil-‐
lea
biebersteinii
Afan.
(Asteraceae)
by
using
Ames/Salmonella
and
E.
coli
WP2
bacterial
test
systems
[1].
The
structural
elucidations
of
the
isolated
compounds
were
done
based
on
physi-‐
cal
observations
and
2D
NMR
experiments.
The
known
mutagens
sodium
azide
(NaN3)
for
S.
typhimurium
TA1535,
9-‐aminoacridine
(9-‐AA)
for
S.
typhimurium
TA1537,
and
N-‐methyl-‐N’-‐
nitro-‐N-‐nitrosoguanidine
(MNNG)
for
E.
coli
WP2
were
used
as
positive
controls
and
10%
DMSO
was
used
as
negative
control
in
these
studies.
As
a
result
of
the
present
study,
two
sus-‐
ceptible
compounds
for
antimutagenic
activity,
quercetin-‐7-‐O-‐β-‐D-‐glucoside
and
patuletin-‐7-‐
O-‐β-‐D-‐glucoside,
were
isolated
from
the
aerial
parts
of
the
plant.
Both
compounds
did
not
show
any
antimutagenic
activity
against
NaN3
and
9-‐AA
induced
mutagenicity.
However,
both
of
these
compounds
clearly
inhibited
MNNG
induced
mutagenesis
in
the
tester
strain
E.
coli
WP2
in
a
dose-‐dependent
manner.
Quercetin-‐7-‐O-‐β-‐D-‐glucoside
inhibited
MNNG
activity
by
causing
decreases
in
the
mutagenicity
at
28.9%
for
4
µM/plate
and
38.2%
for
5
µM/plate.
Similarly,
patuletin-‐7-‐O-‐β-‐D-‐glucoside
showed
a
parallel
activity
at
26.8%
for
4
µM/plate
and
29.9%
for
5
µM/plate
as
well
as
being
non-‐mutagenic
on
the
all
tester
strains
at
the
same
con-‐
centrations.
25-‐40%
inhibition
was
defined
as
moderate
antimutagenicity;
40%
or
more
inhi-‐
bition
as
strong
antimutagenicity;
and
25%
or
less
inhibition
as
no
antimutagenicity
[2].
In
conclusion,
this
is
the
first
detailed
report
that
focuses
on
isolation
and
identification
of
anti-‐
mutagenic
constituents
from
A.
biebersteinii
and
the
present
findings
highlight
its
potential
as
a
source
for
antimutagens.
Keywords:
AMES/Salmonella
test,
antimutagenicity,
E.
coli
WP2
test,
mutagenicity,
patuletin,
quercetin.
References:
[1] Mortelmans
K,
Zeiger
E.
The
Ames
Salmonella/microsome
mutagenicity
assay.
Mutat
Res
2000;
55:
29–60
[2] Güllüce
M,
Agar
G,
Baris
O,
Karadayi
M,
Orhan
F,
Şahin
F.
Mutagenic
and
antimutagenic
effects
of
some
Astragalus
species
grown
in
the
Eastern
Anatolia
Region
of
Turkey.
Phy-‐
tother
Res
2010;
24:
1014–1018
P529
Bauhinia
variegata
L.;
a
bullet
against
pathological
changes
be-‐
hind
cognitive
impairment;
phytochemical,
in
vitro
and
in
vivo
study
Alaa
Selim1,
Heba
Handoussa1,
Nesrine
El-‐Sayed2
1Department
of
Pharmaceutical
Biology,
German
University
in
Cairo,
New
Cairo
City,
11835
Cairo,
Egypt,
2Department
of
Pharmacology
and
Toxicology,
German
University
in
Cairo,
New
Cairo
City,
11835
Cairo,
Egypt
Cognitive impairment, neuroinflammation and oxidative stress are important characteristic
features of neurodegenerative diseases like Alzheimer’s disease. LPS is a well-‐established
model for induction of neuroinflammation and amyloidogenesis widely used to study the
pathway of many neurodegenerative diseases. Phenolics are widely known for their beneficial
effects, they could be considered as promising radiating agents against neuroinflammation. In
this study, hydroalcohol extract of Bauhinia variegata leaves and stalks was shown to amelio-‐
rate neurodegenerative diseases owing to the high phenolic content. Furthermore, the anti-‐
oxidant activity evaluated by cupric reducing antioxidant capacity (CUPRAC) assay was
strongly correlated with total phenolic and flavonoid content within the extract.
To characterize the chemical profile and bioactive composition, twenty phenolic secondary
metabolites were identified using LC-‐qTOF-‐MSn based metabolomics analysis from Bauhinia
variegata leaves and stalks, including flavonoid glycosides and hydroxybenzoic acid deriva-‐
tives. A bioactive compound was identified for the first time from genus Bauhinia, namely
oleuropein, which was further confirmed by spectral means. The effect of Bauhinia variegata
was studied in a dose dependent manner in an LPS induced cognitive impairment model. It
showed significant improvement in neurobehavior even with the least dose studied; 50
mg/kg in both Y-‐maze through enhancing mean % alternation by 57.55% and water maze
through decreasing mean escape latency by 65.19% and increasing preference for platform
area by 1.6 folds. Biochemical testing performed on brain tissues to quantify amyloid beta 42
(Aβ42) by ELISA confirmed the findings in neurobehavioral tests where a significant reducti-‐
on in Aβ42 levels was observed with the used doses as follows: 50 mg/kg by 39.89%, 100
mg/kg by 59.8%, 200 mg/kg by 71.3% and 400 mg/kg by 78.49%. Those outcomes were
compared with Camellia sinensis and donepezil hydrochloride as standard reference drugs.
Keywords: Alzheimer’s disease, Aβ42, Bauhinia variegata, LC-‐qTOF-‐MSn, Phenolics
P530
Production
of
immature
rhizomes
by
breaking
dormancy
under
low
temperature
of
Gastrodia
elata
Blume
Changsu
Kim1,2,
Dongwon
Kim1,
Hyojin
Kim1,
Youngju
Song1,
Wanghyu
Lee2
1
Jeollabukdo
Agricultural
Research
and
Extension
Service,
Iksan
54591,
Korea,
2Plant
Medical
Research
Center, College of Agriculture and Life Science, Chonbuk National University, Jeonju 54896, Korea
Gastrodia
elata
Blume
is
usually
planted
as
immature
rhizomes
in
April
and
harvested
as
ma-‐
ture
rhizomes
in
November
the
following
year.
70%
of
the
total
production
of
G.
elata
in
Ko-‐
rea
is
consumed
raw
and
only
30%
consumed
after
processing.
However,
the
price
of
G.
elata
decrease
greatly
in
November
due
to
seasonal
high
production
at.
The
production
of
G.
elata
should
therefore
preferably
be
constant
all
around
the
year.
Thus,
this
study
was
conducted
to
investigate
the
possibility
of
a
stable
production
of
immature
rhizomes
by
breaking
dormancy
by
low
temperature
treatment.
After
harvesting
G.
elata
in
the
fall,
it
was
planted
in
the
medi-‐
um
(peatmoss
80%
+
perlite
20%,
moisture
content
30
±
2%)
and
kept
at
5
±
1
°C
from
week
2
to
week
10.
Bolting
was
subsequently
induced
at
25
±
1
°C,
and
produced
the
seeds
of
G.
elata
by
artificial
fertilization.
These
seeds
were
induced
to
protocorm
formation
by
Mycena
spp.,
and
immature
rhizomes
produced
by
Armillaria
spp.
[1].
As
the
result,
the
number
of
days
to
emergence
showed
short
as
the
time
of
low
temperature
treatment
period
was
elon-‐
gated,
but
the
emergence
rate,
peduncle
length,
the
number
of
valid
blooming,
the
number
of
valid
pod
increased.
The
natural
fertilization
rate
of
G.
elata
was
0.3%,
whereas
the
artificial
fertilization
rate
increased
with
70.4%
when
it
was
bolting
after
low
temperature
treatment
for
more
than
4
weeks
at
5
°C.
However,
it
was
low
compared
to
that
of
90.4%
when
it
was
harvested
in
natural
condition
of
the
spring
[2].
After
seed
(10,000~20,000
seeds
per
pod)
treated
by
breaking
dormancy
under
low
temperature
condition,
the
protocorm
formation
rate
of
the
germinated
seed
was
from
2.2
to
2.6%
and
its
immature
rhizomes
formation
rate
was
from
8.7
to
9.5%.
Formation
of
protocorm
and
immature
rhizomes
were
proportional
to
the
low
temperature
treatment
period.
As
the
results
of
these
experiments,
even
though
for-‐
mation
ratio
of
protocorm
and
immature
rhizomes
from
G.
elata
harvested
in
the
fall
is
lower
than
in
the
spring,
it
is
considerably
higher
than
the
natural
germination
conditions.
Thus,
a
constant
year-‐round
production
of
G.
elata
would
be
available
due
to
the
application
of
brea-‐
king
dormancy
technology
under
low
temperature
treatment.
Acknowledgements: Rural Development Administration is acknowledged for research fund supporting
Keywords: Gastorida elata Blume, low temperature, breaking dormancy, immature rhizome
References:
[1] Park
EJ,
Lee
WY,
Ahn
JK,
Kim
ST.
A
method
for
in
vitro
production
of
diseases-‐free
Gastro-‐
dia
elata
immature
rhizomes
via
sexual
propagation.
Kor
For
Res
Inst
Sourcebook.
2010.
[2] Hong
IP,
Nam
SH,
Jung
IY,
Sung
GB,
Nam
HW,
Cheong
JC,
Park
JS,
Hur
H,
Lee
MW.
Studies
on
the
conditions
of
seed
germination
of
Gasrodia
elata.
Kor
J
Mycol
2004;
32:
39-‐44.
P531
Gram-‐scale
purification
of
faradiol
from
common
marigold
Dezső
Csupor,1,2
Szabina
Izrael,1
Norbert
Kúsz,1
Attila
Horváth1
1
Department
of
Pharmacognosy,
University
of
Szeged,
Eötvös
u.
6.,
H-‐6720
Szeged,
Hungary,
2
Interdisci-‐
plinary Centre for Natural Products, University of Szeged, Eötvös u. 6., H-‐6720 Szeged, Hungary.
Common
marigold
(Calendula
officinalis
L.)
is
one
of
the
most
widely
used
medicinal
plants
in
the
European
folk
medicine
with
confirmed
anti-‐inflammatory
activity.
The
biological
activity
of
the
flowers
is
related
to
their
triterpenoid
content.
The
pharmacologically
important
triter-‐
penoids
are
alcohols
(monools,
diols
and
triols).
Diols,
such
as
faradiol,
calenduladiol
and
ma-‐
niladiol
and
their
mono-‐
and
diesters
are
characteristic
compounds
of
this
plant
with
remar-‐
kable
antiphlolgistic
activity.
Faradiol
monoester
is
the
most
relevant
principle
for
the
clinical
efficacy
of
the
drug,
due
to
its
quantitative
prevalence.
However,
the
unesterified
faradiol,
which
is
not
present
in
the
extract,
is
the
most
active
member
of
the
Calendula
triterpenoids.
The
anti-‐inflanmmatory
activity
of
marigold
is
related
to
the
inhibition
of
the
proinflammato-‐
ry
cytokines
and
COX-‐2
[2].
Although
the
pharmacological
properties
of
faradiol
make
this
molecule
very
promising
for
further
pharmacological
testing,
phytochemical
studies
have
mainly
focused
on
the
isolation
of
its
ester
derivatives
and
there
are
no
methods
in
the
litera-‐
ture
for
its
quick
and
simple
isolation
from
plant
material
[3].
The
aim
of
our
work
was
to
de-‐
velop
a
method
which
allows
the
gram-‐scale
production
of
faradiol
from
marigold.
Calendula
officinalis
flowers
were
extracted
with
methanol.
The
extract
was
subjected
to
hydrolysis
using
1
M
KOH.
After
precipitation
with
BaCl2,
a
solvent-‐solvent
partitioning
using
CH2Cl2
re-‐
sulted
in
the
faradiol-‐enriched
organic
layer.
Further
separation
was
carried
out
by
MPLC
on
silica
gel,
using
a
gradient
of
cyclohexane
–
diethyl
ether.
The
final
purification,
which
was
performed
by
preparative
HPLC
using
a
C-‐18
stationary
phase
and
a
MeOH
–
H2O
gradient,
led
to
the
isolation
of
pure
faradiol.
The
structure
of
the
isolated
compound
was
confirmed
by
NMR
experiments.
The
yield
of
the
extraction
procedure
was
0,25%,
which
resulted
in
the
isolation
of
1.5
g
pure
faradiol
from
600
g
plant
material.
Keywords:
faradiol,
Calendula
officinalis,
isolation
References:
[1] Preethi
KC,
Kuttan
G,
Kuttan
R.
Anti-‐inflammatory
activity
of
flower
extract
of
Calendula
officinalis
Linn.
and
its
possible
mechanism
of
action.
Indian
J
Exp
Biol
2009;
47:
113-‐120
[2] Hamburger
M,
Adler
S,
Baumann
D,
Förg
A,
Weinreich
B.
Preparative
purification
of
the
major
anti-‐inflammatory
triterpenoid
esters
from
Marigold
(Calendula
officinalis).
Fitot-‐
erapia
2003;
74:
328–338
P532
Anti-‐HIV1
potential
of
an
endemic
Brazilian
bamboo
species
Janayne
Gagliano1
&
Cláudia
Maria
Furlan1
1Department
of
Botany,
Institute
of
Bioscience,
University
of
São
Paulo,
Rua
do
Matão
277,
São
Paulo,
SP
CEP 05508-‐090, Brazil.
Brazil
is
the
country
with
the
highest
diversity
of
bamboo
species
in
the
New
World,
with
34
genera
and
232
native
species
[1].
Knowledge
about
the
medicinal
applications
of
native
bamboos
is
extremely
underdeveloped.
This
study
aimed
at
evaluating
the
anti-‐HIV1
activity
of
extracts
from
leaves
of
Merostachys
pluriflora
Munro
ex.
C.
G.
Camus,
an
endemic
Brazilian
bamboo
from
Atlantic
Rain
Forest.
Plant
material
was
collected,
dried
at
40°C,
powdered
and
macerated
in
70%
ethanol
for
7
days
at
room
temperature
in
the
dark.
The
extract
was
subse-‐
quently
lyophilized,
yielding
the
crude
ethanol
extract
(EE).
EE
was
subjected
to
a
column
chromatography
(CC)
using
silica
gel,
yielding
three
fractions
EE1,
EE2
and
EE3.
EE
and
frac-‐
tions
were
tested
against
the
HIV1
transcriptase
reverse
activity
using
a
colorimetric
kit
(Roche
Diagnostics)
according
to
the
manufacturer's
instructions.
Results
were
expressed
in
minimum
concentration
to
inhibit
50%
of
RT
enzyme
activity
(MIC50).
EE1
showed
the
lower
MIC50
(394.48
µg/mL),
followed
by
EE2
(453.31
µg/mL)
and
EE
(642.61
µg/mL).
Results
sug-‐
gest
that
the
more
polar
fractions
showed
highest
inhibition
of
the
RT
activity.
There
is
no
reports
in
the
literature
of
the
antiviral
potential
of
Brazilian
bamboos,
which
makes
this
a
pioneering
research.
In
this
context,
some
studies
have
demonstrated
that
many
polyphenolic
compounds
act
as
multi-‐target
agents
inhibiting
not
only
the
main
HIV1
replicative
enzyme,
but
also
interfering
on
other
steps
of
the
HIV1
life
cycle
[2].
Bamboos
are
sources
of
C-‐
glycoside
flavonoids,
especially
flavones
derived
from
apigenin,
such
as
vitexin
and
isovitexin
[3].
Some
flavonoids
like
apigenin
can
prevent
HIV1
activation
via
a
mechanism
that
probably
involves
the
viral
transcription
inhibition
[4].
However,
studies
characterizing
M.
pluriflora
polyphenol
composition
and
its
antiviral
activity
are
still
in
progress.
Acknowledgements:
The
authors
thank
CAPES
and
CNPq
for
funding
this
research.
Keywords: antiviral activity, Merostachys, flavone, Atlantic forest.
References:
[1] Filgueiras TS, Gonçalves APS. A Checklist of the basal grasses and bamboos in Brazil (Po-‐
aceae). J Am Bamboo Soc 2004; 18: 7-‐18.
[2] Andrae-‐Marobela
K,
Ghislain
FW,
Okatch
H,
Majinda
RRT.
Polyphenols:
A
diverse
class
of
multi-‐target
anti-‐HIV-‐1
agents.
Curr
Drug
Metab
2013;
14:
392-‐413.
[3] Van
Hoyweghen
L,
De
Beer
T,
Deforce
D,
Heyerick
A.
Phenolic
compounds
and
anti-‐oxidant
capacity
of
twelve
morphologically
heterogeneous
bamboo
species.
Phytochem
Anal
2012;
23:
433–443.
[4] Critchfield
JW,
Butera
ST,
Folks
TM.
Inhibition
of
HIV
activation
in
latently
infected
cells
by
flavonoid
compounds.
AIDS
Res
Hum
Retroviruses
1996;
12:
39–46.
P533
Investigation
of
the
differences
between
the
“Cold”
and
“Hot”
na-‐
ture
on
immune
endocrine
metabolism
levels
of
Açaí
(Euterpe
oleracea
Mart.)
Jianjun
Zhang1,
Linyuan
Wang1*,
Chun
Wang1,
Yingli
Zhu1,
Zichen
Wang1,
Yan
Qu1,
Li
Wu1
1Beijing
University
of
Chinese
Medicine,
Beijing,
100029,
China,
Açaí
(Euterpe
oleracea
Mart.)
has
emerged
as
a
source
of
herbs
in
South
America
[1].
Accord-‐
ing
to
the
theory
of
traditional
Chinese
herbal
properties
we
hypothetically
describe
the
Chi-‐
nese
herbal
properties
of
Açaí
[2].
The
description
and
differentiation
of
the
“Cold”
and
“Hot”
natures
of
Traditional
Chinese
medicine
(TCM)
[3],
have
various
pharmacological
actions
on
serum
metabolites
[4].
In
this
study,
the
divergency
between
the
“Cold”
and
the
“Hot”
natures
was
investigated
through
examining
the
metabolites
of
rats
affected
by
hydrocortisone
(HYD)
and
dexamethasone
sodium
phosphate
(DSP).
In
TCM
theory,
DSP-‐induced
rats
express
“Hot”,
which
were
i.p.
injection
of
DSP
at
the
dose
of
0.35mg/kg
per
day
for
21
days.
While
HYD-‐
induced
rats
express
“Cold”,
which
were
induced
by
i.p.
injection
of
HYD
at
the
dose
of
20
mg/kg
per
day
for
21
days
[5].
Phellodendri
chinensis
cortex
(a
“cold”
TCM)
and
Cortex
cin-‐
namomi
(a
traditional
“hot”
TCM)
served
as
the
positive
control.
Therefore,
the
principle
for
the
treatment
is
to
warm
and
cool
the
organism.
Furthermore,
the
rats
with
DSP-‐induced
or
HYD-‐induced
rats
were
administered
orally,
starting
from
the
7thday,
with
Acaí
at
the
dose
of
1.6
g/kg
and
0.8
g/kg
for
14
days.
Besides,
blood
samples
were
collected
for
the
radioim-‐
munoassay
to
determine
the
levels
of
cyclic
adenosine
monophosphate
(cAMP),
cyclic
guano-‐
sine
monophosphate
(cGMP),
cortisol
(COR),
estradiol
(E2),testosterone
(T),
free
fatty
acids
(FFA),
lipoprotein
lipase
(LPL),
hepatic
lipase
(HL),
Immunoglobulin
G
(IgG),
Immunoglobulin
M
(IgM),
complement
C3(C3),
complement
C4
(C4)
in
two
different
model
were
detected
by
radioimmunoassay.
Our
results
demonstrated
that
Acaí
increased
serum
levels
of
CORT,
T,
IgG,
IgM,
C3
in
DSP-‐induced
rats.
In
addition,
Acaí
decreased
the
levels
of
cGMP,
cAMP,
cAMP/cGMP,
E2,
NEFA,
LPL,
HL
and
C4.
However,
Acaí
have
no
obviously
effect
on
these
tar-‐
gets
in
HYD-‐induced
rats.
Pharmacological
verified
the
Chinese
herbal
properties
“cold”
of
Acaí.
Acknowledgements: Wei Li are acknowledged for technical assistance.
Keywords:
Açaí
(Euterpe
oleracea
Mart.),
“Cold”
and
“Hot”
natures,
hydrocortisone,
dexame-‐
thasone
sodium
phosphate
References:
[1] Ulbricht
C,
Brigham
A,
Burke
D,
Costa
D,
Giese
N,
Lovin
R,
Grimes
Serraano
JM,
Tanguay-‐
Colucci
S,
Weisnerr
W,
Windsor
R.
An
evidence-‐based
systematic
review
of
Acai
(Euterpe
oleracea)
by
the
natural
standard
research
collaboration.
J
Dietary
Suppl
2012;
9:
128-‐
147.
[2] Zhang
J-‐J,
Chen
S-‐H,
Zhu
Y-‐L,
Wang
C,
Wang
J-‐X,
Wang
L-‐Y,
Gao
X-‐M.
Efficacy
analysis
and
theoretical
study
on
Chinese
herbal
properties
of
Acaí
(Euterpe
oleracea).
Zhongguo
Zhong
Yao
Za
Zhi,
2015;
40:
2258-‐2264.
[3] Zhou
C,
Wang
J,
Zhang
X,
Zhao
Y,
Xia
X,
Zhao
H,
Ren
Y,
Xiao
X.
Investigation
of
the
differ-‐
ences
between
the
“COLD”
and
“HOT”
nature
of
Coptis
chinensis
Franch
and
its
processed
materials
based
on
animal’s
temperature
tropism.
Sci
China
C:
Life
Sci
2009;
52:
1073-‐
1080.
[4] Fu
J,
Pang
J,
Zhao
X,
Han
J.
The
quantitative
ideas
and
methods
in
assessment
of
four
prop-‐
erties
of
Chinese
medicinal
herbs.
Cell
Biochem
Biophys
2015;
71:
1307–1312.
[5] Tan
Y,
Liu
X,
Lu
C,
He
X,
Li
J,
Xiao
C,
Jiang
M,
Yang
J,
Zhou
K,
Zhang
Z,
Zhang
W,
Lu
A.
Meta-‐
bolic
profiling
reveals
the
rapeutic
biomarkers
of
processed
Aconitum
Carmichaeli
Debx
in
treating
hydrocortisone
induced
Kidney-‐Yang
deficiency
syndrome
rats.
J
Ethnophar-‐
macol
2014;
152:
585–593.
P534
Evaluation
of
glucosinolate
content
in
radish
(Raphanus
sp.)
germplasm
Ho-‐Cheol
Ko,
Mun-‐Sup
Yoon,
On-‐Sook
Hur,
Jung-‐Sook
Sung,
Jung-‐Bong
Kim,
Jae-‐Gyun
Gwak,
Hyung-‐Jin
Baek,
Sang-‐Gyu
Kim,
Binod
P.
Luitel,
Kyoung-‐Yul
Ryu
and
Ju-‐Hee
Rhee
National
Agrobiodiversity
Center,
National
Institute
of
Agricultural
Science,
Rural
Development
Admini-‐
stration,
370
Nongsaengmyeong-‐ro,
Wansan-‐gu,
54874,
Jeonju,
Republic
of
Korea
Glucosinolate
(GSL)
is
a
secondary
metabolite
and
its
composition
and
content
is
influenced
by
genotype,
climate
and
cultivation
conditions.
Raphanus
species,
including
radish
(Raphanus
sativus
L.),
wild
radish
(R.
raphanistrum)
and
wild
radish
(R.
sativus
subsp.
raphanistroides
)
of
each
32,
4
and
21
accessions
were
analyzed
for
their
GSL
content
using
leaf
and
root
tissues,
respectively.
A
total
of
thirteen
GSLs,
namely
progoitrin,
glucoraphanin,
sinigrin,
glucoalyssin,
gluconapoleiferin,
gluconapin,
4-‐hydroxyglucobrassicin,
glucobrassicanapin,
glucoraphasatin,
glucobrassicin,
4-‐methoxyglucobrassicin,
gluconasturtiin,
and
neoglucobrasscin
were
identi-‐
fied
in
the
both
tissues.
Glucoraphasatin,
glucobrassicin,
and
sinigrin
were
identified
as
the
major
glucosinolates
in
radish
species.
Glucoraphasatin
was
higher
in
root
as
compared
to
leaves
in
all
the
species.
Glucoraphasatin
was
the
highest
in
R.
raphanistrum
(45.9
µmol·g-‐1)
root
followed
by
R.
sativus
subsp.
raphanistroides
(45.1
µmol·g-‐1)
and
the
lowest
(9.7
µmol·g-‐1)
in
R.
raphanistrum
leaves.
Glucobrassicin
was
identified
the
highest
(4.0
µmol·g-‐1)
in
R.
sativus
leaves
followed
by
glucoraphasatin
(3.6
µmol·g-‐1).
Average
total
GSL
was
the
highest
(49.4
µmol·g-‐1)
in
root
of
R.
raphanistrum
followed
by
R.
sativus
subsp.
raphanistroides
(47.6
µmol·g-‐
1).
Total
GSL
was
varied
from
2.8
µmol·g-‐1
(IT102414)
to
81.9
µmol·g-‐1
(K036841)
of
the
root
of
R.
sativus
and
R.
sativus
subsp.
raphanistroides,
respectively
whereas
in
leaves,
it
was
varied
from
1.3
µmol·g-‐1
(K226454)
to
45.1
µmol·g-‐1
(K036846)
of
R.
sativus
and
R.
sativus
subsp.
raphanistroides,
respectively.
Root
tissues
of
R.
raphanistrum
and
R.
sativus
subsp.
rapha-‐
nistroides
exhibited
higher
total
GSL
than
the
leaves
of
Raphanus
species.
This
result
is
useful
to
assist
future
radish
breeding
program
particularly
on
addressing
the
beneficial
glucosinola-‐
te
in
crops.
Acknowledgements: This research was supported by RDA R & D.
Keywords: Glucosinolates, Raphanus species, secondary metabolites, germplasms
References:
[1] Ishida
M,
Hara
M,
Fukino
N,
Kakizaki
T,
Morimitsu
Y.
Glucosinolate
metabolism,
functional-‐
ity
and
breeding
for
the
improvement
of
Brassicaceae
vegetables.
Breed
Sci
2014;
64:
48-‐
59.
P535
Investigation
of
polysaccharide
composition
in
medicinal
and
non-‐medicinal
aloes
Louise
I.
Ahl1,
Henriette
L.
Pedersen2,
William
G.
T.
Willats2,
Nina
Rønsted1,
Olwen
M.
Grace3
1Natural
History
Museum
of
Denmark,
Faculty
of
Science,
University
of
Copenhagen,
Sølvgade
83S,
DK-‐
1307
Copenhagen
K,
Denmark,
2Plant
and
Environmental
Sciences,
Faculty
of
Science,
University
of
Co-‐
penhagen,
Thorvaldsensvej
40,
DK-‐1871
Frederiksberg
C,
Copenhagen,
3Royal
Botanic
Gardens,
Kew,
Sur-‐
rey
TW9
3AE,
United
Kingdom.
The
genus
Aloe
comprises
over
500
species
of
leaf
succulents
found
throughout
Africa,
Mada-‐
gascar
and
the
Arabian
Peninsula.
Aloe
vera
is
a
particularly
widely
known
species,
and
is
used
as
an
ingredient
in
products
from
herbal
medicine
to
cosmetics
and
household
commod-‐
ities.
Aloe
vera
dominates
the
market
worldwide,
but
as
many
as
25%
of
the
Aloe
species
are
used
locally,
a
few
of
them
even
supported
by
small
industries
[1,2].
The
major
chemical
com-‐
ponents
of
the
tissue
used
in
these
products
are
polysaccharides
-‐
highly
complex
molecules
found
in
the
spongy
leaf
mesophyll
(referred
to
as
Aloe
gel).
Studies
of
other
plants
used
in
traditional
medicine
have
identified
bioactive
polysaccharides
and
shown
them
to
have
im-‐
mune-‐modulatory
activity
[3].
In
Aloe,
polysaccharides
extracted
from
the
leaf
mesophyll
have
been
associated
with
both
anti-‐inflammatory
and
immune-‐modulatory
activity
[4].
As
poly-‐
saccharides
are
highly
complex,
they
are
often
studied
indirectly
using
monosaccharide
anal-‐
yses.
A
recent
study
showed
that
the
monosaccharide
composition
of
31
species
of
Aloe
is
conservative,
with
90%
of
the
variation
accounted
for
glucose,
mannose
and
xylose
as
the
ma-‐
jor
constituents
[5].
The
extent
to
which
leaf
mesophyll
polysaccharides
differ
between
Aloe
species
has,
until
now,
remained
unclear.
Here,
we
present
data
from
an
initial
study
of
the
polysaccharide
composition
of
11
species
of
Aloe,
using
carbohydrate
microarrays
and
23
monoclonal
antibodies
binding
to
epitopes
representing
six
distinct
types
of
plant
polysaccha-‐
rides
[6,7].
Marked
variation
in
the
binding
patterns
were
observed
between
the
studied
spe-‐
cies,
indicating
potential
diversity
of
polysaccharides
within
the
genus
Aloe
not
observed
in
analyses
of
the
monosaccharides.
Acknowledgements:
This
work
was
supported
by
the
Villum
Foundation
as
part
of
the
PLANET
project:
Understanding
plant
evolution
and
diversity
in
a
changing
world.
Martin
Aarseth-‐Hansen
and
Paul
Rees
are
acknowledged
for
assistance
with
the
collection
of
plant
material,
and
Paul
Knox
for
the
monoclonal
antibodies.
Keywords:
Aloe,
microarrays,
polysaccharides,
medicinal
plant
use,
gel
References:
[1] Grace
OM,
Simmonds
MSJ,
Smith
MF,
van
Wyk
AE.
Documented
utility
and
biocultural
value
of
Aloe
L.
(Asphodelaceae):
A
review.
Economic
Bot
2009;
63:
167-‐178
[2] Grace
OM.
Current
perspectives
on
the
economic
botany
of
the
genus
Aloe
L.
(Xanthor-‐
rhoeaceae).
S
Afr
J
Bot
2011;
77:
980-‐987
[3] Paulsen
BS,
Barsett
H.
Bioactive
pectic
polysaccharides.
Adv
Polym
Sci
2005;
186:
69-‐101
[4] Steenkamp
V,
Stewart
MJ.
Medicinal
applications
and
toxicological
activities
of
Aloe
prod-‐
ucts.
Pharm
Biol
2007;
45:
411-‐420
[5] Grace
OM,
Dzajic
A,
Jäger
AK,
Nyberg
NT,
Önder
A,
Rønsted
N.
Monosaccharide
analysis
of
succulent
leaf
tissue
in
Aloe.
Phytochemistry
2013;
93:
79-‐87
[6] Moller
I,
Sørensen
I,
Bernal
AJ,
Blaukopf
C,
Lee
K,
Øbro
J,
Pettolino
F,
Roberts
A,
Mikkelsen
JD,
Knox
JP,
Bacic
A,
Willats
WGT.
High-‐throughput
mapping
of
cell-‐wall
polymers
within
and
between
plants
using
novel
microarrays.
The
Plant
Journal
2007;
50:
1118-‐1128
[7] Fagel
JU,
Pedersen
HL,
Melgosa
SV,
Ahl
LI,
Salmean
AA,
Egelund
J,
Rydahl
MG,
Clausen
MH,
Willats
WGT.
Carbohydrate
microarrays
in
plant
science.
Methods
Mol
Biol
2012;
918:
351-‐362
P536
New
insights
into
anti-‐S.
aureus
action
of
Buchenavia
tetraphylla
and
Libidibia
ferrea:
inhibition
of
DNA
replication
Luís
Cláudio
Nascimento
da
Silva1,2,
José
Robson
Neves
Cavalcanti
Filho3,
Clovis
Macedo
Be-‐
zerra
Filho3,
Tiago
Fonseca
da
Silva3,
Márcia
Vanusa
da
Silva3,
Maria
Tereza
dos
Santos
Cor-‐
reia3,
Anders
Løbner-‐Olesen1.
1Department
of
Biology,
University
of
Copenhagen,
Ole
Maaløes
Vej
5
2200
Copenhagen,
Denmark,
2
Post-‐
Graduate
Program
in
Parasite
Biology,
University
of
CEUMA,
Rua
Josué
Montello
1
65.075-‐120,
São
Luís,
Brazil,
3Department
of
Biochemistry,
Federal
University
of
Pernambuco,
Avenida
Professor
Moraes
Rêgo
50670-‐420,
Recife,
Brazil.
DNA
replication
is
an
essential
process
carried
out
by
protein
machinery
with
β-‐clamp
(DnaN)
playing
a
crucial
role,
which
make
it
an
attractive
target
for
potential
new
antibiotics
with
high
therapeutic
index
[1].
This
work
evaluated
the
ability
of
30
extracts
from
22
plants
to
prevent
the
dimerization
of
β-‐sliding
clamp
(DnaN)
of
S.
aureus.
The
antimicrobial
activity
was
evaluated
against
strain
S.
aureus
8325-‐4.
DnaN-‐DnaN
interaction
was
performed
using
a
Bacterial
two-‐hybrid
system
(BTH)
and
confirmed
using
S.
aureus
8325-‐4
derivative
strain
overexpressing
DnaN
and
flow
cytometry
[1].
The
effects
of
SOS
response
was
evaluated
using
a
S.
aureus
strain
with
integrated
recA::lacZ
reporter
[1].
The
combinatory
effects
with
drugs
(ciprofloxacin,
ampicilin
and
chloramphenicol)
and
the
mutagenic
potential
were
also
deter-‐
mined.
The
initial
antimicrobial
screening
revealed
that
eight
extracts
were
able
to
inhibit
S.
aureus,
but
only
three
extracts
inhibited
the
in
vivo
interaction
of
DnaN-‐DnaN:
methanolic
(BTME)
and
ethyl
acetate
(BTEE)extracts
from
Buchenavia
tetraphylla
leaves,
and
aqueous
extract
from
Libidibia
ferrea
fruits
(LFAE).
The
antimicrobial
activity
of
these
two
plants
has
been
reported
by
our
group
[2,3].
Overexpression
of
DnaN
in
S.
aureus
resulted
in
resistance
towards
all
these
plant
extracts.
The
active
extracts
induced
an
increase
in
cell
mass
without
increase
their
DNA
content,
as
expected
for
DNA
replication
inhibitors.
LFAE
and
BTME
showed
synergistic
and
additive
effects
with
chlorampenicol,
respectively;
and
noninteractive
effects
with
the
other
drugs.
BTEE
showed
additive
effects
with
all
tested
drugs.
The
extracts
had
mutagenic
frequencies
smaller
than
ciprofloxacin.
This
work
provides
more
details
about
the
anti-‐S.
aureus
actions
of
B.
tetraphylla
and
L.
ferrea
which
have
DNA
replication
as
poten-‐
tial
target.
The
purification
and
structural
characterization
of
active
compounds
from
all
these
plants
are
the
next
step
of
our
research.
Acknowledgements: Program Science without borders CAPES/Brazil
Keywords: DnaN, Caatinga, natural products
References:
[1] Kjelstrup
S,
Hansen
PMP,
Thomsen
LE,
Hansen
PR,
Løbner-‐Olesen
A.
Cyclic
peptide
inhib-‐
itors
of
the
β-‐sliding
clamp
in
Staphylococcus
aureus.
PloS
one
2013;
8:
e72273.
[2] Oliveira
YLC,
Nascimento
da
Silva
LC,
Silva
AG,
Macedo
AJ,
Araújo
JMD,
Correia
MTS,
Silva
MV.
Antimicrobial
activity
and
phytochemical
screening
of
Buchenavia
tetraphylla
(Aubl.)
RA
Howard
(Combretaceae:
Combretoideae).
Sci
World
J
2012;
2012:
849302.
[3] Silva
LCN,
Sandes
JM,
Paiva
MM,
Araújo
JM,
Figueiredo
RCBQ,
Silva
MV,
Correia
MTS.
Anti-‐
Staphylococcus
aureus
action
of
three
Caatinga
fruits
evaluated
by
electron
microsco-‐
py.
Nat
Prod
Res
2013;
27:
1492-‐1496.
P537
Steroidal
saponins
from
Chlorophytum
deistelianum
Turibio
Tabopda1,2,
Anne-‐Claire
Mitaine-‐Offer1,
Thomas
Paululat3,
Stéphanie
Delemasure4,
Patrick
Dutartre4,
Bonaventure
Tchaleu
Ngadjui2,
Marie-‐Aleth
Lacaille-‐Dubois1
1Laboratoire
de
Pharmacognosie,
EA
4267
FDE,
Université
de
Bourgogne
Franche-‐Comté,
7,
Bd.
Jeanne
d’Arc,
BP
87900,
21079
Dijon
Cedex,
France,
2Département
de
Chimie
Organique,
Université
de
Yaoundé
1,
BP
812
Yaoundé,
Cameroun,
3Universität
Siegen,
OC-‐II,
Naturwissenschaftlich-‐Technische
Fakultät,
Adolf-‐Reichwein-‐Strasse,
D-‐57076
Siegen,
Germany,
4Cohiro,
UFR
des
Sciences
de
Santé,
7,
Bd.
Jeanne
d'Arc,
BP
87900,
21079
Dijon
Cedex,
France
Plants
of
the
genus
Chlorophytum
are
known
as
a
rich
source
of
steroidal
saponins[1-‐3]
which
have
attracted
the
attention
for
their
structural
diversity
and
significant
bioactivities
such
as
cytotoxic,
immunomodulating,
antifungal,
insecticidal
activities,
just
to
mention
a
few
[4].
In
our
continuing
search
for
bioactive
saponins
from
the
genus
Chlorophytum[1-‐3]
we
investi-‐
gated
Chlorophytum
deistelianum
Engl.
&
Krause
(=
C.
sparsiflorum
Backer
var.
sparsiflorum)
from
a
phytopharmacological
point
of
view.
C.
deistelianum
is
a
fleshy
herb
to
50
cm
high,
with
white
or
greenish
flowers.
No
study
on
the
secondary
metabolites
of
this
plant
has
been
re-‐
ported
so
far.
Therefore,
we
report
herein
the
isolation
of
four
previously
undescribed
steroi-‐
dal
saponins,
called
chlorodeistelianosides
A
–
D
(1-‐4)
together
with
five
known
steroidal
saponins
from
C.
desteilianum.
Their
structures
were
determined
by
spectroscopic
methods
including
1D
and
2D
NMR
experiments,
ESI
and
HRESIMS.
Compounds
1
and
2
are
glycosides
of
hecogenin
and
its
24β-‐OH
derivative,
respectively,
whereas
3
is
a
glycoside
of
(25R)-‐5α-‐
22α-‐methoxyfurostane-‐2α,3β,26
triol
and
4
a
glycoside
of
(25R)-‐5α-‐furost-‐20(22)-‐en-‐12-‐
one-‐3β,26
diol.
Compounds
2-‐4
share
the
same
sugar
sequence
at
C-‐3
characterized
as
β-‐D-‐
glucopyranosyl-‐(1→2)-‐[β-‐D-‐xylopyranosyl-‐(1→3)]-‐β-‐D-‐glucopyranosyl-‐(1→4)-‐β-‐D-‐
galactopyranosyl
whereas
the
sugar
sequence
of
1
was
elucidated
as
β-‐D-‐glucopyranosyl-‐
(1→3)-‐[α-‐L-‐rhamnopyranosyl-‐(1→4)]-‐β-‐D-‐xylopyranosyl-‐(1→3)-‐[β-‐D-‐glucopyranosyl-‐
(1→2)]-‐β-‐D-‐glucopyranosyl-‐(1→4)-‐β-‐D-‐galactopyranosyl.
Furthermore,
seven
compounds
were
examined
for
cytotoxicity
against
one
human
colorectal
adenocarcinoma
cell
line
(SW480)
and
one
rat
cardiomyoblast
cell
line
(H9c2).
Among
them,
three
known
spirostane-‐
type
glycosides
exhibited
cytotoxicity
on
both
cell
lines
with
IC50
ranging
from
8
to
10
µM.
Keywords: Chlorophytum deistelianum, Asparagaceae, steroid saponins, NMR, cytotoxicity
References:
[1] Acharya
D,
Mitaine-‐Offer
A-‐C,
Kaushik
N,
Miyamoto
T,
Paululat
T,
Lacaille-‐Dubois
M-‐A.
Fu-‐
rostane-‐type
steroidal
saponins
from
the
roots
of
Chlorophytum
borivilianum.
Helv
Chim
Acta
2008;
91:
2262–2269
[2] Acharya
D,
Mitaine-‐Offer
A-‐C,
Kaushik
N,
Miyamoto
T,
Paululat
T,
Mirjolet
JF,
Duchamp
O,
Lacaille-‐Dubois
M-‐A.
Cytotoxic
spirostane-‐type
saponins
from
the
roots
of
Chlorophytum
borivilianum.
J
Nat
Prod
2009;
72:
177–181
[3] Acharya
D,
Mitaine-‐Offer
A-‐C,
Kaushik
N,
Miyamoto
T,
Paululat
T,
Mirjolet
JF,
Duchamp
O,
Lacaille-‐Dubois
M-‐A.
Steroidal
saponins
from
Chlorophytum
orchidastrum.
J
Nat
Prod
2010;
73:
7–11.
[4] Lacaille-‐Dubois
M-‐A.
Bioactive
saponins
with
cancer
related
and
immunomodulatory
ac-‐
tivity:
recent
developments.
In:
Atta-‐Ur-‐Rahman,
editor.
Studies
in
Natural
Products
Chemistry
series.
Amsterdam:
Elsevier,
2005;
32:
209–246
P538
The
use
of
medicinal
plants
grown
hydroponically
in
the
treat-‐
ment
of
wounds
Maxim
Timoshenko1
,
Natalya
Voroshilova1
,
Marlen
Yessirkepov1,
Assilbek
Burabaev1.
1
Department
of
Biology,
Biochemistry
and
Microbiology,
South
Kazakhstan
State
Pharmaceutical
Acad-‐
emy, Al-‐Farabi Square, 160019, Shymkent, Republic of Kazakhstan
The
deleterious
changes
in
environmental
conditions
such
as
water
stress
bring
physiological
and
biochemical
changes
in
plants,
which
results
in
crop
loss
[1].
Hydroponics
-‐
is
a
method
of
growing
plants
in
which
the
plants
receive
nutrients
from
a
special
solution.
Hydroponics
has
the
following
advantages:
1)
the
maximum
realization
of
the
genetic
potential
of
plants,
2)
the
crop
can
increase
to
20-‐30%,
3)
a
positive
impact
on
the
environment,
and
4)
rational
use
of
natural
resources.
The
objective
of
this
study
was
to
investigate
the
efficacy
of
medicinal
plants
grown
hydroponically
in
the
treatment
of
wounds.
We
chose
chamomile
as
a
model
plant
for
this
study.
Chamomile
is
a
medicinal
plant,
which
presents
several
biological
effects,
especially
the
anti-‐inflammatory
effect.
One
of
the
compounds
related
to
this
effect
is
apig-‐
enin,
a
flavonoid
that
is
mostly
found
in
its
glycosylated
form,
apigenin-‐7-‐glucoside
(APG),
in
natural
sources
[1].
Chamomile
was
grown
by
the
standard
method
and
hydroponically.
Ex-‐
tracts
of
both
plants
were
obtained.
52
rabbits
of
such
breeds
as
"White
Giant"
were
used
as
laboratory
animals.
The
average
age
of
animals:
2
years,
the
average
weight:
3.7
kg.
Under
local
anaesthesia,
we
made
a
5-‐7
cm
long
incision
on
both
sides.
The
wound
was
sutured
with
conventional
circular
vertical
seams.
The
right-‐side
wound
(the
control
group),
was
treated
with
a
chamomile
extract
of
chamomile
grown
by
the
standard
method.
The
left-‐side
wound
was
treated
with
chamomile
extract
of
chamomile
grown
hydroponically.
The
efficiency
of
healing
was
assessed
on
parameters
such
as:
density,
elasticity,
scar
height
over
the
wound
and
colour.
Each
parameter
had
four
degrees
and
were
evaluated
on
a
scale
from
0
to
3.
0
points
meant
the
absence
of
expression
of
a
characteristic,
3
points
meant
the
most
pro-‐
nounced
characteristic.
The
maximum
amount
of
points:
12.
Results
were
recorded
every
day
for
7
days.
The
wound-‐healing
efficiency
of
the
control
group
was
below
the
efficiency
of
the
healing
of
wounds
that
were
treated
with
chamomile
extract,
grown
hydroponically.
The
dif-‐
ference
between
the
sum
of
the
points
of
the
experimental
group
and
the
sum
of
the
points
of
the
control
group
on
the
first
day
was
1.7
point,
the
last
day
3.8
point.
Conclusion:
Chamomile
extract,
which
was
grown
hydroponically,
is
effective
in
the
treatment
of
wounds.
Acknowledgements: Bekaidar Nurmashev is acknowledged for the support of the administration
Keywords: Hydroponics, medicinal plants, chamomile, treatment of wounds.
References:
[1] Tripathi
P,
Rabara
RC,
Shulaev
V,
Shen
QJ,
Rushton
PJ.
Understanding
water-‐stress
re-‐
sponses
in
soybean
using
hydroponics
system
-‐
A
systems
biology
perspective.
Front
Plant
Sci
2015;
6:
1145.
[2] Miguel
FG,
Cavalheiro
AH,
Spinola
NF,
Ribeiro
DL.
Validation
of
a
RP-‐HPLC-‐DAD
method
for
chamomile
(Matricaria
recutita)
preparations
and
assessment
of
the
marker,
apig-‐
enin-‐7-‐glucoside,
safety
and
anti-‐inflammatory
effect.
Evid
Based
Complement
Altern
Med
2015;
3:
2015:828437.
[3] Bagirov
V.,
Lyakina
M.
Rules
of
building
quality
standards
tinctures
homoeopathic
matrix.
Pharmacy
2003;
6:
37-‐39.
P539
Usefulness
of
Φ-‐order
kinetics
for
the
investigation,
characterisa-‐
tion
and
quantification
of
photodegradation
processes.
Mounir
Maafi,
Mohammed
Alqarni
Leicester School of Pharmacy, De Montfort University, The Gateway, Leicester LE1 9BH, UK
Φ-‐order
kinetics
has
been
shown
to
be
the
best
tool
to
study
photokinetics
[1-‐6].
Various
spe-‐
cies
have
been
studied
using
this
approach
including
photochromes
[1,
2]
and
drugs
[3-‐6].
The
unimolecular
photodegradation
processes
of
the
form
AB
(1Φ)
and
photoreversible
sy-‐
stems
AB
(2Φ),
where
A,
the
initial
species
phototransforms
into
a
product
B
(A→B)
which
itself
can
return
to
A
(A→B)
via
two
distinct
photoprocesses,
each
characterized
each
by
an
individual
efficiency
(Φ),
have
been
mathematically
described
by
integrated
rate-‐law
equati-‐
ons
which
express
the
so
called
'Φ-‐order'
kinetics
[1-‐6].
The
application
of
this
novel
ap-‐
proach
was
found
to
be
useful
to
determine
a
number
of
photoreaction
attributes.
The
reac-‐
tion's
photochemical
quantum
yield(s),
the
effects
of
initial
species
concentration,
or
the
im-‐
pact
of
competitive
absorbers
on
the
rate-‐constant
of
the
photoactive
species
were
all
made
readily
accessible
by
simple
methods.
More
importantly,
the
Φ-‐order
kinetics
has
proven
to
facilitate
the
development
of
a
wide
range
of
actinometers
from
AB
(1Φ)
and
AB
(2Φ)
systems
[2-‐6].
This
novel
Φ-‐order
kinetic
method
clearly
has
the
potential
for
application
in
various
fields
including
the
photodegradation
of
natural
products.
The
details
of
Φ-‐order
elucidation
kinetic
methods
will
be
discussed
here
and
examples
of
application
presented.
Acknowledgements:
The
authors
would
like
to
thank
Taif
University,
Saudi
Arabia
and
the
Saudi
cultural
Bureau
(in
London),
for
their
help,
assistance
and
financial
support.
Keywords: Spectrophotokinetics, photodegradation; Φ-‐order kinetics; quantum yield; acti-‐
nometry; UV/Vis spectroscopy
References
[1] Maafi
M,
Brown
RG.
The
kinetic
model
for
AB
(1
Φ)
systems:
A
closed-‐form
integration
of
the
differential
equation
with
a
variable
photokinetic
factor.
J
Photochem
Photobiol
A:
Chemistry
2007;
187:
319-‐324.
[2] Maafi
M.
The
potential
of
AB
(1Φ)
systems
for
direct
actinometry.
Diarylethenes
as
suc-‐
cessful
actinometers
for
the
visible
range.
Phys
Chem
Chem
Phys
201012:
13248-‐13254.
[3] Maafi
W,
Maafi
M.
Modelling
nifedipine
photodegradation,
photostability
and
actinome-‐
tric
properties.
International
journal
of
pharmaceutics.
2013;456(1):153-‐64.
[4] Maafi
M,
Maafi
W.
Ф-‐order
kinetics
of
photoreversible-‐drug
reactions.
Int
J
Pharm.
2014;
471:
536-‐543.
[5] Maafi
M,
Maafi
W.
Montelukast
photodegradation:
Elucidation
of
Ф-‐order
kinetics,
de-‐
termination
of
quantum
yields
and
application
to
actinometry.
Int
J
Pharm
2014;
471:
544-‐552.
[6] Maafi
M,
Lee
LY.
Determination
of
Dacarbazine
Φ-‐Order
Photokinetics,
Quantum
Yields,
and
Potential
for
Actinometry.
J
Pharm
Sci
2015;
104:
3501-‐3509.
P540
Mechanism(s)
of
action
involved
in
the
antimycobacterial
activity
of
Tetracera
macrophylla
Siti
Fatimah
Sabran1,2,
Maryati
Mohamed1,2,
Mohd
Fadzelly
Abu
Bakar1,2,
Alona
Cuevas
Lina-‐
toc1,2
1Centre
of
Research
for
Sustainable
Uses
of
Natural
Resources,
Faculty
of
Science,
Technology
and
Human
Development,
Universiti
Tun
Hussein
Onn
Malaysia,
86400
Parit
Raja,
Batu
Pahat,
Johor,
Malaysia,
2Department
of
Technology
and
Heritage,
Faculty
of
Science,
Technology
and
Human
Development,
Uni-‐
versiti Tun Hussein Onn Malaysia, 86400 Parit Raja, Batu Pahat, Johor, Malaysia
Tetracera
macrophylla
Wall.
ex
Hook.
f.
&
Thoms.
(Dilleniaceae)
is
locally
known
as
akar
mempelas.
The
stems
have
been
used
traditionally
by
the
Jakun
tribe
in
Johor,
Peninsular
Ma-‐
laysia
to
treat
night
fever,
which
is
a
specific
symptom
of
tuberculosis
(TB).
This
study
aimed
to
investigate
the
mechanism(s)
of
antimycobacterial
action
of
ethyl
acetate
extract
of
T.
mac-‐
rophylla
stems
(ETM)
against
Mycobacterium
smegmatis
(ATCC
700084),
an
in
vitro
model
strain
of
TB.
Potential
mechanisms
of
action
were
investigated
by
observing
the
effects
of
ETM
on
growth
of
bacterial
cells
upon
treatment
using
time-‐kill
assay
[1],
determination
of
cell
membrane
integrity
by
measuring
the
supernatant
of
M.
smegmatis
absorbing
at
280
nm
(OD280)
[2],
changes
in
cellular
morphology
using
field
emission-‐scanning
electron
microscopy
(FE-‐SEM)
[3],
and
differences
of
protein
expression
by
2D
gel
electrophoresis
analysis
[4,5].
Major
phytochemical
constituents
were
also
profiled
by
GC-‐MS
analysis.
Results
showed
that
the
ETM
significantly
(p<0.05)
reduced
over
99.9%
bacterial
cells
within
8
hours
of
treatment
at
4-‐fold
minimum
inhibitory
concentration
(MIC=1.56
mg/mL).
When
the
ETM
was
added
at
1-‐fold
MIC,
it
significantly
(p<0.05)
deteriorated
the
integrity
of
cell
membrane,
causing
cellu-‐
lar
leakage
and
extreme
cell
damage.
There
were
43
proteins
significantly
(p<0.05)
expressed
differentially
after
ETM
treatment,
including
17
up-‐regulated
proteins
and
26
down-‐regulated
proteins.
More
than
50%
of
the
ETM
comprised
of
pyrazole
(26.8%)
and
phenanthroline
(25.6%)
derivatives,
which
might
contribute
to
the
antimycobacterial
activity.
In
conclusion,
the
ETM
exerted
antimycobacterial
action
via
several
mechanisms
and
T.
macrophylla
could
be
a
potential
lead
of
anti-‐TB
natural-‐based
drugs
worthy
of
further
investigation.
Acknowledgements:
Johor
Biotechnology
and
Biodiversity
Corporation
(J-‐Biotech),
Johor
National
Parks
Corporation
(JNPC),
Universiti
Putra
Malaysia
(UPM),
Ministry
of
Higher
Education
of
Malaysia
(MOHE),
and
Fundamental
Research
Grant
Scheme
Project
No.
1435
Keywords: Traditional knowledge, medicinal plant, phytochemical, tuberculosis
References
[1] Zhao
L,
Zhang
H,
Hao
T,
Li
S.
In
vitro
antibacterial
activities
and
mechanism
of
sugar
fatty
acid
esters
against
five
food-‐related
bacteria.
Food
Chem
2015,
187:
370-‐377.
[2] Kim
S,
Lee
H,
Lee
S,
Yoon
Y,
Choi
K-‐H.
Antimicrobial
Action
of
Oleanolic
Acid
on
Listeria
monocytogenes,
Enterococcus
faecium,
and
Enterococcus
faecalis.
PLoS
One
2015;
10:
e0118800.
[3] Piroeva
I,
Dimowa
L,
Sbirkova
H.
A
simple
and
rapid
scanning
electron
microscope
pre-‐
parative
technique
for
observation
of
biological
samples :
application
on
bacteria
and
DNA
samples.
Bulg
Chem
Commun
2013;
45:
510-‐515.
[4] O’Farrell
PH.
High
resolution
two
dimensional
electrophoresis
of
proteins.
J
Biol
Chem
1975,
250:
4007-‐4021.
[5] Burgess-‐Cassler
A,
Johansen
JJ,
Santek
DA,
Ide
JR,
Kendrick
NC.
Computerized
quantitative
analysis
of
Coomassie-‐Blue-‐stained
serum
proteins
separated
by
two-‐dimensional
elec-‐
trophoresis.
Clin
Chem
1989;
35:
2297-‐2304.
P541
Bioassay-‐guided
isolation
studies
on
Scutellaria
salviifolia
Benth.
Zeynep
Dogan,
V.
Murat
Kutluay,
Iclal
Saracoglu
Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Hacettepe
University,
06100,
Sihhiye,
Ankara,
Tur-‐
key
Scutellaria
salviifolia
Benth.
(Lamiaceae)
is
an
endemic
plant
of
Turkey
[1].
Previous
studies
of
this
genus
have
reported
the
isolation
of
flavonoids,
iridoid
glucosides,
phenylethanoid
glyco-‐
sides,
diterpenoids,
triterpenoids,
alkaloids
and
essential
oils.
Species
and
their
active
princi-‐
ples
possess
antioxidant,
antitumor,
antiangiogenesis,
hepatoprotective,
anticonvulsant,
anti-‐
bacterial,
and
antiviral
activities
[2].
Free
radicals
are
one
of
the
key
factors
that
cause
several
disorders,
including
cancer.
For
these
reason,
investigations
on
the
anticancer
activity
related
with
antioxidant
compounds
are
getting
very
important
[3].
On
the
other
hand,
recent
discov-‐
eries
found
liver
X
receptors
(LXRs)
could
regulate
tumor
growth
in
a
variety
of
cancer
cell
lines
[4].
In
the
present
study,
we
aimed
to
isolate
active
compounds
of
S.
salviifolia
on
the
basis
of
free
radical
scavenging
capacity
against
DPPH,
NO
and
SO
radicals,
cytotoxic
activity
against
HEp-‐2
cell
line,
and
LXRs
ligand
activity
using
a
LXRα
luciferase
reporter
assay.
Aque-‐
ous
extract
of
S.
salviifolia
showed
potent
SO
radical
scavenging
(IC50
159
µg/mL),
LXRα
ago-‐
nistic
(nearly
equivalent
to
activity
of
10
nM
T0901317,
positive
control,
at
100
µg/mL)
and
cytotoxic
(IC50
250
µg/mL)
activities.
The
extract
was
subjected
to
polyamide
column
chroma-‐
tography
to
afford
five
main
fractions
(Frs.
A-‐E).
Phenolic
compounds-‐rich
fractions
(Frs.
D,
E)
also
showed
significant
activities
in
the
both
test
systems
with
the
IC50
ranges
of
20-‐202
µg/mL
for
radicals,
116
and
33
µg/mL
for
Hep-‐2
cell
line,
respectively.
Repeated
column
chromatographies
of
the
Frs.
D
and
E
resulted
in
the
isolation
of
four
phenolic
compounds
in
pure
form.
Structures
of
the
isolated
compounds
were
identified
as
3,5-‐dihydroxyphenyl-‐6'-‐O-‐
E-‐p-‐coumaroyl-‐β-‐glucopyranoside
(1),
hispidulin
(2),
luteolin-‐4'-‐O-‐β-‐glucopyranoside
(3),
and
apigenin-‐5-‐O-‐β-‐glucopyranoside
(4)
by
means
of
1D-‐
and
2D-‐NMR
data.
Studies
on
active
constituents
are
still
in
progress.
Acknowledgements:
This
study
was
supported
by
The
Scientific
and
Technological
Research
Council
of
Turkey
(Program
No:
2211-‐C)
and
Hacettepe
University
BAB
(Project
No:
014-‐D08
301
001).
The
authors
are
grateful
to
Prof.
Dr.
Toshiaki
Makino
and
Dr.
Ishiuchi
Kan’ichiro
(Nagoya
City
University,
Faculty
of
Pharmaceutical
Sciences,
Nagoya,
Japan)
for
recording
NMR
spectra.
Keywords:
Scutellaria
salviifolia,
free
radical
scavenging,
cytotoxic
activity,
phenolic
com-‐
pounds.
References:
[1] Davis
PH.
Flora
of
Turkey
and
East
Aegean
Islands.
Volume
7.
Edinburg:
University
Press,
1982,
78-‐100.
[2] Shang
X,
He
X,
He
X,
Li
M,
Zhang
R,
Fan
P,
Zhang
Q,
Jia
Z.
The
genus
Scutellaria
an
eth-‐
nopharmacological
and
phytochemical
review.
J
Ethnopharmacol
2010,
128:
279-‐313.
[3] Harput
US,
Genc
Y,
Saracoglu
I.
Cytotoxic
and
antioxidative
activities
of
Plantago
lagopus
L.
and
characterization
of
its
bioactive
compounds.
Food
Chem
Toxicol
2012,
50:
1554-‐
1559.
[4] Zhang
W,
Jiang
H,
Zhang
J,
Zhang
Y,
Liu
A,
Zhao
Y,
Zhu
X,
Lin
Z,
Yuan
X.
Liver
X
receptor
activation
induces
apoptosis
of
melanoma
cell
through
caspase
pathway.
Cancer
Cell
Int
2014,
14:
16.
P542
Compounds
from
the
aerial
parts
of
Agrimonia
pilosa
and
their
protein
tyrosine
phosphatase
1B
and
acetylcholinesterase
inhibi-‐
tory
activities
Duc
H.
Nguyen1,2,
U
M.
Seo1,
Duc
D.
Le1,
Thi
T.
Nguyen1,
Jae
Sue
Choi,3
Mi
H.
Woo
1
1College
of
Pharmacy,
Catholic
University
of
Daegu,
Gyeongsan
38430,
Republic
of
Korea.
2Phutho
College
of
Pharmacy,
Viettri
City,
Phutho
Province
290000,
Vietnam.
3Department
of
Food Science & Nutrition, Pukyong National University, Busan 48547, Republic of Korea
Agrimonia
pilosa,
a
perennial
herb,
belongs
to
the
Rosaceae
family.
Whole
plant
of
A.
pilosa
has
been
used
traditionally
as
hemostatic,
antimalarial,
and
antidysenteric
in
Chinese
herbal
medi-‐
cine
for
a
long
time
[1,2].
Two
new
flavanonol
glucoside
isomers,
(2R,3S)
dihydrokaempferol
3-‐O-‐β-‐ᴅ-‐glucoside
(9)
and
(2S,3R)
dihydrokaempferol
3-‐O-‐β-‐ᴅ-‐glucoside
(10),
were
isolated
from
the
aerial
parts
of
Agrimonia
Pilosa,
along
with
12
known
compounds
(1‒8,
11‒14).
The-‐
ir
structures
were
determined
on
the
basis
of
spectroscopic
analyses.
In
addition,
all
the
isola-‐
tes
were
evaluated
for
PTP1B
inhibitory
activity.
Among
them,
compounds
13
and
14
display-‐
ed
potent
inhibitory
activity
against
PTP1B
with
IC50
values
of
7.14
±
1.75
and
7.73
±
0.24
μM,
respectively.
Furthermore,
all
the
isolated
compounds
were
found
to
inhibit
AChE
with
IC50
values
ranging
from
62.96
±
0.35
to
118.32
±
0.09
μM.
Therefore,
chemical
constituents
of
the
aerial
parts
of
A.
pilosa
have
a
beneficial
effect
for
the
treatment
of
type
2
diabetes,
obesities,
and
Alzheimer’s
disease.
Keywords:
Agrimonia
pilosa,
flavanonol
glucosides,
protein
tyrosine
phosphatase
1B,
ace-‐
tylcholinesterase
References:
[1] Kim
JJ,
Jiang
J,
Shim
DW,
Kwon
SC,
Kim
TJ,
Ye
SK,
Kim
MK,
Shin
YK,
Koppula
S,
Kang
TB,
Choi
DK,
Lee
KH.
Anti-‐inflammatory
and
anti-‐allergic
effects
of
Agrimonia
pilosa
Ledeb
extract
on
murine
cell
lines
and
VOA-‐induced
airway
inflammation.
J
Ethnopharmacol
2012;
140:
213-‐221.
[2] Taira
J,
Nanbu
H,
Ueda
K.
Nitric
oxide-‐scavenging
compounds
in
Agrimonia
pilosa
Ledeb
on
LPS-‐induced
RAW264.7
macrophages.
Food
Chem
2009;
115:
1221-‐1227.
P543
Evaluation
of
5-‐HT1A
and
5-‐HT2A
receptors
in
a
rat
model
of
chronic
restraint
stress:
antidepressant-‐like
effects
of
albiflorin
Yingli
Zhu1,
Linyuan
Wang1,
Jiangxia
Wang1,
Zhihui
Yang2,
Chenglong
Wang1,
Danping
Zhao1,
Jianjun
Zhang1*
1Beijing
University
of
Chinese
Medicine,
Beijing,
100029,
China,2Department
of
Psychiatry
&
Neurosci-‐
ence, University of Florida, Gainesville, FL 32611, USA.
Albiflorin
(AF),
a
monoterpene
glycoside
isolated
from
Paeoniae
Radix
Alba,
is
widely
used
in
Chinese
herbal
medicine
[1].
However,
its
effect
on
psychiatric
disorders
has
not
been
well
studied.
This
study
aimed
to
evaluate
the
anti-‐depressant-‐like
effects
of
AF
in
a
rat
model
of
chronic
restraint
stress
(CRS)
and
to
explore
the
possible
mechanisms.
CRS
was
induced
in
the
stress
group
by
a
daily
6h-‐restraining
in
a
plastic
restrainer
for
continuous
21
days
[2].
Rats
were
given
30
mg/kg
or
15
mg/kg
of
AF
for
3
weeks.
Fluoxetine
(2.5
mg/kg)
served
as
a
positive
control.
Furthermore,
body
weight
measurement
and
behavioural
tests
were
applied,
including
the
sucrose
preference
test,
open
field
test
and
the
elevated
plus-‐maze.
Moreover,
measurements
of
the
levels
of
neurochemical
(hypothalamic–pituitary–adrenal,
HPA
axis),
monoamine,
serotonin
receptors
(5-‐HT1AR
/5-‐HT2AR)
in
the
hippocampus
by
ELISA,
RT-‐PCR
or
Western
Blot
were
involved
in
the
mechanism
studies
[3].
Our
results
demonstrated
that
AF
increased
body
weight,
sucrose
solution
consumption
and
restored
the
depressant-‐like
behaviors.
Besides,
AF
also
significantly
increased
the
levels
of
5-‐HT,
5-‐hydroxyindoleacetic
acid
(5-‐HIAA),
noradrenaline
(NE),
dopamine
(DA)
and
5-‐HT1AR
mRNA
and
protein
expres-‐
sion.
In
addition,
AF
decreased
the
levels
of
corticotropin
releasing
hormone
(CRH),
adreno-‐
corticotropic
hormone
(ACTH),
cortisol
(CORT)
and
inhibited
the
overexpression
of
5-‐HT2AR
mRNA
and
protein
expression.
These
results
suggested
that
AF
produced
antidepressant-‐like
effects
via
regulating
hippocampal
serotonergic
systems.
Acknowledgements:
The
authors
are
grateful
for
financial
support
from
National
Nature
Science
Foun-‐
dation
of
China
(Project
NO.
81473370)
and
Patent
(No.
ZL
201110184287.4,
China)
Keywords: Albiflorin, chronic restraint stress, antidepressant, serotonergic systems
References:
[1] National
Committee
of
Pharmacopoeia,
2015.
Pharmacopoeia
of
the
People’s
Republic
of
China
(Part
one).
Chemical
Industry
Press,
Beijing,
p.
105.
[2] Liu
L,
Zhou
X,
Zhang
Y,
Liu
Y,
Yang
L,
Pu
J,
Zhu
D,
Zhou
C,
Xie
P.
The
identification
of
meta-‐
bolic
disturbances
in
the
prefrontal
cortex
of
the
chronic
restraint
stress
rat
model
of
de-‐
pression.
Behav
Brain
Res
2016;
305:
148-‐156.
[3] Muguruza
C,
Miranda-‐Azpiazu
P,
Díez-‐Alarcia
R,
Morentin
B,
Gonzalez-‐Maeso
J,
Callado
LF,
Meana
JJ.
Evaluation
of
5-‐HT2A
and
mGlu2/3
receptors
in
postmortem
prefrontal
cortex
of
subjects
with
major
depressive
disorder:
Effect
of
antidepressant
treatment.
Neuro-‐
pharmacol
2014;
86:
311-‐318.
P544
Inhibitory
effects
of
epimedium
herb
water
extract
on
the
in-‐
flammatory
response
in
vitro
and
in
vivo
You-‐Chang
Oh1,
Yun
Hee
Jeong1,
Won-‐Kyung
Cho1,
Sang-‐Joon
Lee2,
and
Jin
Yeul
Ma1
1KM
Application
Center,
Korea
Institute
of
Oriental
Medicine,
70
Chemdan-‐ro,
Dong-‐gu,
Daegu,
701-‐300,
Daegu,
South
Korea,
2Toxicity
Screening
Research
Center
Gyeongnam
Department
of
Environmental
Tox-‐
icology
and
Chemistry,
Korea
Institute
of
Toxicology,
17,
Jegok-‐gil,
Munsan-‐eup,
Jinju-‐si,
Gyeongsangnam-‐
do,
660-‐844,
South
Korea
Epimedium
Herb
(EH)
is
a
traditional
herbal
drug
used
for
the
treat
genital
pain,
benign
pros-‐
tatic
hyperplasia,
low
back
pain,
memory
impairment
in
the
middle-‐aged
population,
and
in-‐
flammatory
diseases,
such
as
lymphadenopathy
and
genital
inflammation
in
eastern
Asia.
Al-‐
so,
EH
previously
reported
about
several
biological
activities
related
to
cancer,
injury
and
asthma
[1-‐3].
In
the
study
described
herein,
we
investigated
the
biological
effects
of
Epimedi-‐
um
Herb
water
extract
(EHWE)
on
LPS-‐mediated
inflammation
in
macrophages
and
local
in-‐
flammation
in
vivo.
We
also
investigated
the
biological
effects
of
EHWE
on
the
production
of
inflammatory
mediators,
pro-‐inflammatory
cytokines
and
related
products,
as
well
as
NF-‐κB
and
MAPK
activation
in
LPS-‐stimulated
macrophages.
The
analgesic
effect
of
the
acetic
acid-‐
induced
writhing
response
and
inhibitory
activity
on
xylene-‐induced
ear
edema
was
also
evaluated
in
mice.
EHWE
exhibited
anti-‐inflammatory
effects
by
inhibiting
the
production
of
NO,
IL-‐6
and
IL-‐1β.
In
addition,
EHWE
strongly
suppressed
iNOS,
a
NO
synthesis
enzyme,
in-‐
duced
HO-‐1
expression,
and
inhibited
NF-‐κB
activation
as
well
as
MAPK
pathway
phosphory-‐
lation.
Furthermore,
EHWE
exhibited
an
analgesic
effect
on
the
writhing
response
and
an
in-‐
hibitory
effect
on
ear
edema
in
mice.
For
the
first
time,
we
demonstrate
the
anti-‐inflammatory
effects
and
inhibitory
mechanism
in
macrophages,
as
well
as
the
inhibitory
activity
of
EHWE
in
vivo.
Our
results
indicate
a
potential
use
of
EHWE
as
an
inflammatory
therapeutic
agent
developed
from
a
natural
substance.
Acknowledgements:
This
work
was
supported
by
the
grant
K16281
awarded
to
Korea
Institute
of
Orien-‐
tal
Medicine
(KIOM)
from
Ministry
of
Education,
Science
and
Technology
(MEST),
Korea.
Keywords: Epimedium herb, heme oxygenase-‐1, nuclear factor-‐kappaB, mitogen-‐activated
protein kinase, anti-‐inflammatory effect, analgesic effect
References:
[1] Xie
JY,
Dong
JC,
Cui
Y.
Effect
of
epimedium
herb
on
RANTES
and
monocyte
chemotactic
protein-‐3
expression
in
lung
tissue
of
asthmatic
rats.
Zhongguo
Zhong
Xi
Yi
Jie
He
Za
Zhi
2008;
28:
238–241.
[2] Tohda,
C.
Nagata,
A.
Epimedium
koreanum
extract
and
its
constituent
icariin
improve
mo-‐
tor
dysfunction
in
spinal
cord
injury.
Evid
Based
Complement
Altern
Med.
2012;
2012:
731208.
[3] Indran
IR,
Zhang
SJ,
Zhang
ZW,
Sun
F,
Gong
Y,
Wang
X,
Li
J,
Erdelmeier
CA,
Koch
E,
Yong
EL.
Selective
estrogen
receptor
modulator
effects
of
epimedium
extracts
on
breast
cancer
and
uterine
growth
in
nude
mice.
Planta
Med
2014;
80:
22–28.
P545
LC-‐MS
profiling,
isolation
and
identification
of
bioactive
second-‐
ary
metabolites
from
a
marine-‐derived
fungus
Arthrinium
sp.
Ahmed
M.
Elissawy1,
Sherif
S.
Ebada1,
Mohamed
L.
Ashour1,
Ferhat
C.
Özkaya2,
Weaam
Ebrahim3,4,
Mohamed
S.
ElNaggar1,
Peter
Proksch4,
Abdelnasser
Singab1
1
Department
of
Pharmacognosy
and
Phytochemistry,
Faculty
of
Pharmacy,
Ain-‐Shams
University,
Cairo
11566,
Egypt,
2
Department
of
Aquaculture,
Faculty
of
Fisheries,
Izmir
Katip
Celebi
University,
Izmir
35100
Bornova,
Turkey,
3
Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Mansoura
University,
Mansoura
35516,
Egypt,
4
Institut
für
Pharmazeutische
Biologie
und
Biotechnologie,
Heinrich-‐Heine-‐
Universität,
40225
Düsseldorf,
Germany
Marine
ecosystems
represent
a
relatively
recent,
rich
and
potential
source
of
bioactive
me-‐
tabolites,
many
of
which
are
being
used
as
drug
products,
others
in
different
phases
of
clinical
trials,
still
many
more
representing
promising
bioactivities
in
the
preclinical
trials
[1].
In
the
last
decade,
special
interests
were
directed
toward
the
investigation
of
endophytic
microor-‐
ganisms
as
a
new
source
of
novel
lead
compounds
with
potential
biological
activities
[2].
LC-‐
MS
guided
fractionation
of
ethyl
acetate
(EtOAC)
extract
of
a
marine-‐derived
fungus
Arthrini-‐
um
sp.
isolated
from
the
inner
tissues
of
Turkish
marine
sponge
Sarcotragus
muscarum
grow-‐
ing
on
solid
rice
medium,
led
to
the
isolation
of
the
new
natural
product
4-‐hydroxyphenyl-‐
ethyl-‐3-‐propanoate
(1),
in
addition
to
eleven
known
compounds
including
alkylphenol
deriv-‐
atives
(2-‐3),
anthraquinones
(4-‐5),
xanthone
derivatives
(6-‐7),
3,4-‐dimethoxy-‐
protocatecheuic
acid
(8),
triterpenes
(9-‐10),
griseofulvin
(11)
and
its
dechloro-‐derivative
(12).
Structures
of
all
isolated
compounds
were
elucidated
on
the
basis
of
extensive
spectro-‐
scopic
analyses
including
1D,
2D
NMR
&
HPLC-‐ESI-‐MS
spectra
supported
by
comparison
with
reported
literature
data.
Keywords:
Marine
derived
fungus,
Arthrinium,
xanthones,
anthraquinones,
alkylphenol
de-‐
rivatives
Reference:
[1] Zhang
G,
Li
J,
Zhu
T,
Gu
Q,
Li
D.
Advanced
tools
in
marine
natural
drug
discovery.
Curr
Opin
Biotechnol
2016;
42:
13-‐23
[2] Elissawy
AM,
El-‐Shazly
M,
Ebada
SS,
Singab
AB,
Proksch
P.
Bioactive
terpenes
from
marine
derived
fungi.
Mar
Drugs
2015;
13:
1966-‐1992
P546
Marine-‐derived
Aspergillus
species
are
promising
source
of
new
secondary
metabolome
Ala
Ud
Din1,
Wayne
D.
Inman2,
Frederick
A.
Valeriote3,
Phillip
Crews2
1
Department
of
Chemistry,
Bacha
Khan
University
Charsadda
25120,
Pakistan,
2
Department
of
Chemis-‐
try
and
Biochemistry,
University
of
California,
Santa
Cruz,
California
95064,
3
Josephine
Ford
Cancer
Cen-‐
ter,
Henry
Ford
Hospital,
Detroit,
Michigan
48202
Hawaiian shallow water sediment derived-‐fungi; Aspergillus ochraceopetaliformis (088702A)
and Aspergillus insulicola (088708A) resulted into isolation of two new cyclotetrapeptides
Ochrine A and B, and a novel hexacyclic dipeptide Azonazine[1] from their culture broth after
impressive cytotoxic data of its crude extracts. The structures Ochrine A and B were found to
be cyclo[-‐L-‐lle-‐L-‐Thr-‐L-‐O-‐Me-‐Tyr-‐L-‐pro-‐] and cyclo[-‐L-‐lle-‐L-‐Thr-‐L-‐O-‐Me-‐Tyr-‐L-‐N-‐Me-‐Ala-‐] re-‐
spectively. The sequence and absolute configurations of amino acids were determined by ex-‐
tensive NMR, ESI-‐ MS fragmentation and Marfey’s analysis [2].
Acknowledgements: This research was supported by NIH grants CA 47135 (PC), CA 052955 (PC) and
support from Higher Education Commission (HEC) of Pakistan under IRSIP program.
Keywords: Marine-‐derived-‐fungi, cyclotetrapeptides, Marfey’s analysis, Anti-‐tumor activity
References:
[1] Wu
QX,
Crews
MS,
Draskovic
M,
Sohn
J,
Johnson
TA,
Tenney
K,
Valeriote
FA,
Yao
XJ,
Bjel-‐
danes
LF,
Crews
P.
Azonazine,
a
novel
dipeptide
from
a
hawaiian
marine
sediment-‐
derived
fungus,
aspergillus
insulicola.
Org
Lett
2010;
12:
4458–4461
[2] Fujii
KY,
Ikai
T,
Suzuki
M,
Oka
MS,
Harada
KI.
A
nonempirical
method
using
LC/MS
for
de-‐
termination
of
the
absolute
configuration
of
constituent
amino
acids
in
a
peptide:
combi-‐
nation
of
Marfey’s
method
with
mass
spectrometry
and
its
practical
application.
Anal
Chem
1997;
69:
5146–5151
P547
Analgesic
activity
of
Carthamus
caeruleus
ethanolic
extracts
Amel
Toubane,
Kamel
Daoud
Laboratory
of
transfer
phenomena,
University
of
Science
and
Technology
Houari
Boumediene,
PB32,
El
Alia,
Bab
Ezzouar,
16111,
Algiers,
Algeria
Pain
is
a
symptom
that
is
associated
with
many
diseases
especially
inflammatory
diseases
and
burns
[1].
Carthamus
caeruleus
L.
[Asteraceae]
is
an
Algerian
plant
that
is
traditionally
used
for
burns
healing
and
has
shown
an
anti-‐inflammatory
activity.
The
aim
of
this
study
was
to
investigate
the
analgesic
activity
of
C.
caeruleus,
which
was
collected
from
the
east
of
the
city
of
Algiers
(Dellys),
Algeria.
Roots
were
cut
and
dried
in
a
dark
room
and
used
later
for
a
con-‐
ventional
solvent
extraction
(with
optimized
conditions
of
heat
power
and
extraction
time)
using
ethanol
as
solvent
to
obtain
the
crude
extract.
The
peripheral
analgesic
activity
of
the
crude
extract
was
assessed
using
the
method
of
acetic
acid
induced
writhing
in
mice,
two
treatments
of
crude
extract,
aspirin
(reference)
and
physiologic
water
(control)
were
admin-‐
istrated
respectively
at
the
doses
of
246
mg/kg,
446
mg/kg,
300
mg/kg
and
10ml/kg,
30
min
prior
to
acetic
acid
(1%)
intraperitonial
injection
at
the
dose
of
10ml/kg.
Acetic
acid
releases
endogenous
mediators
that
stimulate
the
nociceptive
neurons
creating
an
abdominal
con-‐
striction
response
[2].
The
ethanolic
extracts
of
C.
caeruleus
with
the
dose
of
446
mg/
kg
body
weight
decreases
the
number
of
mice
writhing
(37,5%)
with
a
significant
difference
from
the
control,
the
result
of
the
treatment
is
comparable
to
what
was
obtained
with
the
reference
aspirin
(38,8%)
with
no
significant
difference
between
them.
The
antinociceptive
action
of
the
extract
may
be
due
to
the
inhibition
of
the
released
endogenous
mediators
[3].
Carthamus
caeruleus
root
extract
present
a
potential
as
analgesic
drug
for
the
treatment
of
pain
with
a
dose
dependent
effect.
Acknowledgements:
Research
and
Development
Center
of
the
national
pharmaceutical
company
SAIDAL
is
acknowledged.
Keywords: Carthamus caerueleus, analgesic activity, writhing inhibition
References:
[1] Bhaskar
V
H,
Balakrishnan
N.
Analgesic,
anti-‐inflammatory
and
antipyretic
activities
of
Pergularia
daemia
and
Carissa
carandas.
DARU
J
Pharmaceut
2015;
17:
168−174
[2] Lalerinzuali
K,
Vabeiryureilai
M,
Jagetia
GJ.
Investigation
of
the
anti-‐inflammatory
and
analgesic
activities
of
ethanol
extract
of
stem
bark
of
Sonopatha
Oroxylum
indicum
in
vi-‐
vo.
Int
J
Inflam
2016;
2016:
1−8
[3] Sengar
N,
Joshi
A,
Prasad
SK,
Hemalatha
S.
Anti-‐inflammatory,
analgesic
and
antipyretic
activities
of
standardized
root
extract
of
Jasminum
sambac.
J
Ethnopharmacol
2015;
160:
140−148
P548
Biological
active
natural
products
from
marine
organisms
and
terrestrial
plants
of
Iran
Amir
Reza
Jassbi
Medicinal
and
Natural
Products
Chemistry
Research
Center,
Shiraz
University
of
Medical
Sciences,
Shiraz,
Iran.
Email:
jassbiar@sums.ac.ir
Bioassay-‐guided
fractionation
and
purification
of
different
terrestrial
plants
or
marine
organ-‐
isms,
like
sponges
and
algae,
from
the
Persian
Gulf
resulted
in
identification
of
various
natural
products
with
anti-‐cancer,
antibacterial
and
antioxidant
activities
[1].
Three
steroids
and
one
furanosesquiterpenoid
have
been
isolated
from
a
dichloromethane
extract
of
a
sponge,
Ircinia
mutans
collected
from
the
shallow
waters
of
Larak
Island.
The
extracts
and
isolated
com-‐
pounds
were
tested
against
three
human
cancer
cell
lines;
HT-‐29
(human
colorectal
adeno-‐
carcinoma),
MCF-‐7
(human
breast
adenocarcinoma)
and
MOLT-‐4
(lymphoblastic
leukemia)
in
a
3-‐(4,5-‐dimethylthiazol-‐2-‐yl)-‐2,5-‐diphenyltetrazolium
bromide
(MTT)
colorimetric
cytotoxic
assay.
Exploring
the
cytotoxic
activity
of
several
red
and
brown
algae,
including
Dichotomaria
obtusata
yielded
tocopherols
and
phytol
esters
as
cytotoxic
metabolites
against
LS180
(hu-‐
man
colon
adenocarcinoma);
MCF-‐7
and
MOLT-‐4
cell
lines.
Some
20
plant
species
from
family
Solanaceae,
Lamiaceae
and
Anthemideae
such
as
endemic
members
of
Solanum,
Hyoscyamus,
Anthemis,
Salvia,
Thymus,
and
Ballota
were
screened
for
their
cytotoxic
activity
against
MOLT-‐
4
cell
lines
[2,3].
The
most
bioactive
species
were
determined
as
dichloromethane
extracts
of
A.
susanna,
A.
Gayana
and
A.
odontostephana
and
the
roots
of
S.
lachnocalyx
with
IC50s
of
6.7±0.0,
5.6±0.9,
7.5±0.8
and
26.3±11.8
μg/ml,
respectively
against
the
above
mentioned
can-‐
cerous
cell
lines.
The
active
extracts
were
further
subjected
to
the
respective
bioassay
guided
purification
to
afford
their
bioactive
natural
compounds
such
as
abietane
diterpenoids
in
the
roots
of
S.
lachnocalyx,
spathulenol
and
farnesylgeraniol
from
the
shoots
of
this
plant.
The
structures
of
the
isolated
compounds
were
characterized
using
1
and
2
D
NMR
and
mass
spec-‐
tral
data.
The
anticancer
potential
of
the
tested
plants
extracts
were
mainly
determined
to
be
due
to
their
terpene
contents,
among
which
abietane
and
rearranged-‐abietane
diterpenoids
from
Salvia
species
were
the
most
active
ones
against
the
tested
cell
lines,
while
sesquiterpe-‐
noids
were
active
principles
in
the
extracts
of
Anthemis
species
and
the
sponges.
The
tochopherols
also
are
responsible
for
the
cytotoxic
activity
of
the
red
algae,
D.
Obtusata.
Acknowledgements:
I
am
thankful
to
all
of
my
PhD
students,
the
technicians
and
my
colleague
Dr.
O.
Firuzi
for
their
scientific
collaboration
and
Research
Council
of
Shiraz
University
of
Medical
Sciences
for
the
financial
support
of
my
research
activity
Keywords: Salvia, Anthemis, Ircinia, red and brown algae
References:
[1] Jassbi
AR,
Mohabati
M,
Eslami
S,
Sohrabipour
J,
Miri
R.
Biological
activity
and
chemical
constituents
of
red
and
brown
algae
from
the
Persian
Gulf.
Iran
J
Pharm
Res
2013;
12:
339-‐348
[2] Jassbi
AR,
Zare
S,
Firuzi
O,
Xiao
J.
Bioactive
phytochemicals
from
shoots
and
roots
of
Salvia
species.
Phytochem
Rev
2015,
DOI:
10.1007/s11101-‐015-‐9427-‐z
[3] Shokoohinia
Y,
Sajjadi
SE,
Jassbi
AR,
Moradi
H,
Ghassemi
N,
Schneider
B.
Sesquiterpenes
and
Flavonoids
of
Anthemis
odontostephana
var.
odontostephana.
Chem
Nat
Comp
2015;
51:
491-‐494
P549
The
toxicity
of
ribbon
worms:
Alpha-‐nemertides
or
tetrodotoxin,
or
both?
H.S.
Andersson1,
E.
Jacobsson2,
C.
Eriksson2,
M.
Hedström3,
H.
Seth4,
P.
Sundberg4,
K.J.
Rosen-‐
gren5,
M.
Strand
4,6,
U.
Göransson2
1
Linnaeus
University
Centre
for
Biomaterials
Chemistry,
Department
of
Chemistry
and
Biomedical
Sci-‐
ences,
Linnaeus
University,
Norra
vägen
49,
392
34
Kalmar,
Sweden,
2
Division
of
Pharmacognosy,
De-‐
partment
of
Medicinal
Chemistry,
Uppsala
University,
Box
570.
751
23
Uppsala,
Sweden,
3Department
of
Biotechnology,
Lund
University,
Box
118,
221
00
Lund,
Sweden,
4
Department
of
Biological
and
Environ-‐
mental
Sciences,
University
of
Gothenburg,
Box
463,
405
30
Gothenburg,
Sweden,
5
School
of
Biomedical
Sciences,
The
University
of
Queensland,
Brisbane,
QLD
4072,
Australia,
6
Swedish
Species
Information
Centre,
Swedish
University
of
Agricultural
Sciences,
Bäcklösavägen
10,
756
51
Uppsala,
Sweden
The
marine
ribbon
worms
(nemerteans)
are
predators
which
capture
their
prey
by
everting
a
proboscis
carrying
a
mixture
of
toxins
which
brings
on
rapid
paralysis
[1].
Moreover,
ribbon
worms
have
a
thick
layer
of
epidermal
mucus
of
similar
constitution.
Tetrodotoxin
(TTX)
has
been
identified
as
one
of
these
toxins
[2].
The
extreme
toxicity
of
TTX
(lethal
by
ingestion
of
0.5-‐2
mg)
is
due
to
its
ability
to
block
voltage-‐gated
sodium
channels.
Although
several
bacterial
species
(among
these
Vibrio
sp.)
have
been
linked
to
its
synthesis,
the
biogenic
origin
and
biosynthesis
is
unclear.
One
hypothesis
is
that
TTX
production
occurs
in
a
symbiotic
relationship
with
its
host,
in
this
case
the
ribbon
worm
[3].
We
have
made
significant
effort
to
identify
TTX
in
a
setup
for
production
through
the
cultivation
of
Vibrio
alginolyticus
in
nutrient
broth
infused
with
mucus
from
the
ribbon
worm
Lineus
longissimus.
Toxicity
was
demonstrated
by
fraction
injections
into
shore
crabs,
but
no
TTX
was
found,
and
it
could
be
shown
conclusively
that
toxicity
was
unrelated
to
TTX
and
the
Vibrio
culture
itself,
and
rather
a
constituent
of
the
ribbon
worm
mucus
[4].
The
following
studies
led
us
to
the
discovery
of
a
new
class
of
peptides,
the
alpha-‐nemertides,
in
the
mucus
of
the
ribbon
worms,
which
could
be
directly
linked
to
the
toxic
effects.
A
literature
review
of
the
available
evidence
for
TTX
in
ribbon
worms
show
that
the
evidence
in
most
cases
are
indirect,
although
notable
exceptions
exist.
This
points
to
the
necessity
to
further
investigate
the
presence
and
roles
of
TTX
and
alpha-‐nemertides
in
ribbon
worms.
References:
[1] Kem
WR.
Structure
and
activity
of
Nemertine
Toxins.
Integrative
and
Comparative
Biology
1985;
25:
99-‐111
[2] Miyazawa
K,
Higashiyama
M,
Ito
K,
Noguchi
T,
Arakawa
O,
Shida
Y,
Hashimoto
K.
Tetrodotoxin
in
two
species
of
ribbon
worm
(Nemertini),
Lineus
fuscoviridis
and
Tubulanus
punctatus.
Toxicon
1988;
26:
867-‐874
[3] Carroll
S,
McEvoy
EG,
Gibson
R.
The
production
of
tetrodotoxin-‐like
substances
by
nemer-‐
tean
worms
in
conjunction
with
bacteria.
Journal
of
Experimental
Marine
Biology
and
Ecology
2003;
288:
51-‐63
P550
Mo-‐clay
for
treatment
of
psoriasis
Kirstine M Hansen, Anna K Jäger, Åsa Andersson
Department
of
Drug
Design
and
Pharmacology,
Faculty
of
Health
and
Medical
Sciences,
University
of
Copenhagen,
Universitetsparken
2,
DK-‐2100,
Copenhagen,
Denmark
Mo-‐clay
was
used
by
German
doctors
to
treat
injured
soldiers’
wounds
during
the
First
World
War.
Today,
there
are
anecdotal
cases
of
mo-‐clay
being
beneficial
for
patients
suffering
from
psoriasis,
a
chronic,
inflammatory
disease.
There
are
several
histological
features
in
the
psori-‐
atic
skin,
including
acanthosis,
hyperkeratosis,
pararkeratosis
and
a
loss
of
granular
layer.
Mo-‐clay
is
a
unique
marine
deposit,
an
Eocene
clayed
diatomite.
It
was
formed
54
million
years
ago
from
deposits
of
single-‐celled
algae
along
with
clay
minerals
and
volcanic
ash.
The
major
elements
are
silicon,
aluminium
and
iron.
It
is
found
in
Denmark
and
Germany.
As
mo-‐
clay
had
been
used
to
treat
wounds,
it
was
tested
for
antibacterial
activity.
Mo-‐clay
did
not
show
any
anti-‐bacterial
activity
against
a
battery
of
Gram-‐positive
and
–negative
bacteria.
Mo-‐
clay
showed
stimulation
of
cell
proliferation
at
concentrations
39-‐78
μg/ml
in
splenic
mouse
lymphocytes,
and
at
156
μg/ml
in
HaCat
cells,
whereas
an
inhibition
of
proliferation
was
ob-‐
served
at
313
μg/ml.
Mo-‐clay
was
tested
for
anti-‐psoriatic
activity
in
vivo
using
the
mouse
tail
test
[1].
This
model
can
be
used
to
investigate
agents
for
effect
on
psoriasis,
since
the
adult
mouse
tail
has
regions
of
both
orthokeratosis
and
parakeratosis.
Mo-‐clay
induced
or-‐
thokeratosis
and
showed
a
significant
increase
in
epidermis
thickness.
The
results
suggest
that
mo-‐clay
may
have
anti-‐psoriatic
effects.
Keywords: Mo-‐clay, psoriasis, mouse tail test
References:
[1] Bosman
B,
Matthiesen
T,
Hess
V,
Friderichs
E.
A
Quantitative
method
for
measuring
antipsoriatic
activity
of
drugs
by
the
mouse
tail
test.
Skin
Pharmacol
Physiol
1992;
5:
41-‐
48
P551
Determination
of
the
antimicrobial
and
antitussive
activities
of
the
leaves
of
Solanum
torvum
Abena
Amponsaa
Brobbey1,
Anna
Kwarley
Quartey2,
Samuel
Otuo-‐Serebour1,
Isaac
Ayensu1
1
Department
of
Pharmaceutical
Chemistry,
Faculty
of
Pharmacy
&
Pharmaceutical
Sciences,
College
of
Health
Sciences,
Kwame
Nkrumah
University
of
Science
and
Technology,
Kumasi,
Ghana,
2
Department
of
Pharmaceutical
Sciences,
School
of
Applied
Sciences,
Central
University
College,
Accra,
Ghana
Cough
is
a
protective
reflex
which
prevents
the
entry
of
foreign
materials
into
the
respiratory
tract
and
also
aids
the
expulsion
of
mucus.
The
nature
of
coughing
may
depict
the
presence
of
a
disease.
Cough
may
be
caused
by
an
infection
of
the
respiratory
tract
by
viruses
and
bacteria
[1].
Solanum
torvum
(Swartz)
is
a
valuable
medicinal
plant
of
the
Solanaceae
family.
Various
parts
of
this
plant
are
used
in
the
treatment
of
different
diseases
such
as
cough,
asthma,
dia-‐
betes,
microbial
infections
and
liver
diseases
just
to
mention
a
few
[2].
This
study
screen
for
phytochemicals,
antimicrobial
and
antitussive
effects
on
the
methanol
extract
of
the
leaves
of
S.
torvum.
Extraction
of
the
leaves
was
done
by
cold
macerating
with
methanol,
phytochemical
screening
was
done
using
methods
described
by
Ayensu
&
Quartey
[3],
Minimum
Inhibitory
Concentration
(MIC)
of
the
leave
extract
was
obtained
using
agar
well
diffusion
method
at
concentrations
5.0-‐0.625
mg/mL
screened
against
selected
micro-‐organisms
implicated
in
cough
(Staphylococcus
aureus,
Streptococcus
pyogenes,
Klebsiella
pneumonia
and
Escherichia
coli).
Antitussive
activity
was
accessed
using
citric
acid
method.
Phytochemical
screening
on
the
leaves
extract
showed
the
presence
of,
tannins,
flavonoids,
reducing
sugars,
saponin
gly-‐
cosides,
alkaloids,
phytosteroids
and
terpenoids.
MIC
obtained
for
the
antimicrobial
assay
were
2.5
mg/ml,
3.16
mg/ml,
5.0
mg/ml
and
5.0
mg/ml
for
S.
aureus,
S.
pyogenens,
K.
pneumo-‐
nia
and
E.
coli
respectively.
The
antitussive
activity
of
the
extract
showed
a
dose
dependent
activity
giving
percentage
cough
inhibition
of
53.4
%,
64.3
%
and
73.6
%
for
doses
of
100
mg/kg,
300
mg/kg
and
1000
mg/kg
respectively
with
76.2
%
for
the
positive
control
dihydro-‐
codeine
and
17.4
%
for
the
negative
control
normal
saline.
The
methanol
extract
of
the
leaves
of
S.
torvum
have
important
phytochemical
constituents,
some
antimicrobial
activity
and
a
dose
dependent
antitussive
activity.
Acknowledgements:
Gabriel
Dapaah
is
acknowledged
for
technical
assistance.
Keywords:
Solanum
torvum,
antitussive
activity,
antimicrobial
activity,
dihydrocodeine,
citric
acid
References:
[1] Reynolds
S,
Mackenzie
A,
Spina
D,
Page
C.
The
pharmacology
of
cough
trends.
Pharmacol
Sci
2004;
25:
569-‐576
[2] Jaiswal
B.
Solanum
torvum,
A
Review
of
its
traditional
uses,
phytochemistry
and
pharma-‐
cology.
Int
J
Pharm
Bio
Sci
2012;
3;
104-‐111
[3] Ayensu
I,
Quartey
A.
Phytochemical
screening
and
in-‐vitro
antioxidant
properties
of
the
stem
bark
of
Trichilia
tessmannii
(Harms),
World
J
Pharm
Pharmaceut
Sci
2015;
4:
76-‐90
P552
Commercializing
traditional
knowledge:
Regulating
access
and
benefit-‐sharing
in
Namibia
Anne Heeren1, Ahmad Cheikyoussef2, Percy Chimwamurombe3
1Faculty
of
Architecture
and
Landscape
Planning,
University
of
Hannover,
Herrenhäuser
Straße
8,
30419
Hannover,
Germany,
2Science
&
Technology
Division,
Multidisciplinary
Research
Centre,
University
of
Namibia,
340
Mandume
Ndemufayo
Ave,
13301
Windhoek,
Namibia.
3Department
of
Biological
Sciences,
University
of
Namibia,
340
Mandume
Ndemufayo
Ave,
13301
Windhoek,
Namibia.
In
biodiversity-‐rich
countries
the
creation
of
new
markets
to
finance
sustainable
development
has
become
the
central
premise
of
today's
environmental
policy-‐making.
In
Namibia
access
to
traditional
knowledge
(TK)
to
be
employed
in
commercial
product
development
is
going
to
be
regulated
under
the
forthcoming
access
and
benefit-‐sharing
(ABS)
regulation.
The
basic
principle
of
ABS
is
the
establishment
of
the
state's
sovereignty
over
its
genetic
resources
(and
associated
TK).
Access
rights
for
the
intangible
information
attached
to
plants
are
established.
A
market
for
TK
is
being
created
where
users
and
providers
negotiate
over
the
conditions
of
exchange.
As
a
result,
ABS
agreements
are
reached
where
access
to
TK
is
granted
in
exchange
to
specific
compensation
measures,
e.g.
capacity
building
and
technology
transfer.
In
Namibia
there
is
a
controversial
debate
on
how
to
regulate
the
use
of
TK:
In
order
to
access
resources
to
be
used
in
R&D
user
promote
simple,
streamlined
and
easy
to
regulate
ABS
procedures
[1].
In
contrast,
knowledge
holder
stress
further
cultural
and
societal
issues
related
to
plant
uses.
This
may
lead
to
fundamentally
contrary
expectations
on
how
to
adequately
address
negotiation
procedures
and
the
allocation
of
benefits
[2].
In
this
study
I
will
present
two
Namibian
bioprospecting
cases:
Hoodia
(Hoodia
gordonii)
[3]
and
marama
bean
(Tylosema
esculentum)
[4].
The
main
research
question
is
how
“nature”
is
being
negotiated
by
the
diverse
actors
involved.
This
is
done
by
highlighting
the
Societal-‐Relations-‐
to-‐Nature
(SRN),
the
entanglement
of
both
physical-‐material
and
cultural-‐symbolic
aspects
of
commercially
used
indigenous
knowledge
systems
(IKS).
Findings
can
be
used
to
develop
best
practices
on
participatory
procedures
in
the
context
of
the
implementation
of
ABS
mechanisms.
Keywords:
Access
and
Benefit-‐Sharing,
Biodiversity,
Traditional
Knowledge,
Indigenous
Knowledge
Systems,
Societal
Relations
to
Nature
References:
[1] Shikongo
ST.
The
issue
of
bio-‐trade
and
bio-‐prospecting
in
Namibia:
an
analytical
overview.
Marrakech
ABS
Workshop,
Marrakech
2011
[2] Vermeylen
S.
From
life
force
to
slimming
aid:
exploring
views
on
the
commodification
of
traditional
medicinal
knowledge.
Appl
Geography
2008;
28:
224-‐235
[3] Wynberg
R,
Schroeder
D,
Chennels
R.
Indigenous
peoples,
consent
and
benefit
sharing:
lessons
from
the
San-‐Hoodia
case.
London:
Springer;
2009
[4] Chimwamurombe
P,
Mapaure
I,
Claassen
P.
Understanding
the
relationship
between
indigenous
(traditional)
knowledge
systems
(IKS)
and
access
to
genetic
resources
and
benefits
sharing
(ABS).
Afr
J
Biotechnol
2010;
9:
9204-‐9207
P553
Oxymatrine
and
matrine
biotransformation
by
microorganisms
and
the
evaluation
of
effects
on
5-‐lipoxygenase
inhibition
Fatih
Demirci1,2,
Ayşe
Esra
Karadağ3,4,
Mustafa
Güzel5
,
1
Anadolu
University,
Faculty
of
Pharmacy,
Department
of
Pharmacognosy
2
Faculty
of
Health
Sciences,
26470-‐Eskişehir,
3
Graduate
School
of
Health
Sciences,
4
Medipol
University,
Faculty
of
Pharmacy,
De-‐
partment
of
Pharmacognosy;
5
Faculty
of
Internal
Medicine,
Department
of
Medical
Pharmacology,
34810,
Kavacık/Beykoz,
İstanbul,
Turkey
Sophora
L.
species
of
Fabaceae
contain
bioactive
quinolizidine
type
alkaloids
such
as
ox-‐
ymatrine
and
matrine.
It
is
well
documented
that
some
Sophora
species
are
used
in
Tradition-‐
al
Chinese
Medicines.
The
mechanisms
of
inflammation
involve
complex
cascades
of
events
where
chemical
messengers
play
an
important
role.
An
important
class
of
messengers
are
the
pro-‐inflammatory
leukotrienes
resulting
from
the
metabolism
of
arachidonic
acid,
which
be-‐
gins
with
its
oxidation
by
the
enzyme
5-‐lipoxygenase.1
The
aim
of
this
study
was
to
synthesize
derivatives
of
oxymatrine
and
matrine
by
microbial
biotransformation.
As
a
result,
oxymatrine
was
metabolized
to
its
deoxy-‐analogue
matrine,
by
Fusarium
solani,
Saccharomyces
cerevisiae,
Mucor
ramannianus,
Alternaria
alternata
and
Pyc-‐
noporus
cinnabarinus
among
15
different
microorganisms
evaluated.
The
metabolites
were
evaluated
by
LC-‐MS/MS.
In
the
time-‐based
evaluation
of
the
transformation,
complete
metab-‐
olization
of
oxymatrine
was
observed
after
48
hours
of
incubation.
The
substrate
matrine
was
metabolized
into
two
different
analogues
by
M.
ramannianus.
Main
metabolite
was
tentatively
assigned
as
hydroxy-‐derivative
of
matrine,
where
the
other
was
identified
as
an
aromatized
derivative.
Additionally,
both
the
substrate
and
metabolites
were
evaluated
in
vitro
for
their
lipoxygenase
(5-‐LOX)
inhibitory
activity
by
means
of
spectrophotometric
methods.
As
a
result,
oxymatrine
showed
an
inhibition
of
21.1%,
where
matrine
exhibited
10.5%,
respectively.
The
metabolites
showed
no
inhibition
at
the
tested
concentrations.
To
the
best
of
our
knowledge
this
is
the
first
biotransformation
of
oxymatrine
and
matrine
by
the
above
mentioned
microorganisms
and
LOX
evaluation.
The
biological
evaluation
of
matrine
derivatives
is
ongoing.
Acknowledgement:
This
work,
which
is
part
of
the
MSc
thesis
of
AEK
was
supported
by
Anadolu
Universi-‐
ty,
Scientific
Research
Projects
Commission
Fund
Projects
grant
no:
1601S029
Keywords: Matrine, oxymatrine, microbial biotransformation, LC-‐MS/MS, lipoxygenase in-‐
hibitor activity
References:
[1] Baylac
S,
RacineP.
Inhibition
of
5-‐lipoxygenase
by
essential
oils
and
other
natural
fra-‐
grant
extracts.
Int
J
Aromather
2003;
13:
138-‐142
P554
Chemical
and
genetic
differences
between
Hawaiian
lineages
of
the
alga
Asparagopsis
taxiformis
Benjamin
R.
Clark1,
2,
Mindy
Mizobe3,
Jo-‐Ann
Leong3,
Robert
P.
Borris1,
2
1
College
of
Pharmacy,
University
of
Hawaii
at
Hilo,
HI
96720,
USA,
2Current
address:
School
of
Pharma-‐
ceutical
Science
and
Technology,
Tianjin
University,
Tianjin,
300072,
P.
R.
China,
3
Hawai‘i
Institute
of
Marine
Biology,
University
of
Hawaii,
Kaneohe,
Hawaii
96744,
USA
Asparagopsis
taxiformis
is
a
widely-‐distributed
marine
alga
found
throughout
tropical
regions
[1].
In
Hawai’i
it
is
known
as
limu
kohu,
and
is
prized
as
a
foodstuff
for
its
peppery
flavour.
The
chemistry
of
A.
taxiformis
has
been
well-‐studied,
and
is
dominated
by
bromoform
and
a
range
of
small
halogenated
ketones,
acids,
and
esters
[2].
In
2004,
sequencing
of
mitochondrial
DNA
revealed
distinctive
genetic
differences
between
different
populations
of
A.
taxiformis
[3],
while
a
2008
study
indicated
that
three
of
these
lineages
were
present
in
Hawaiian
waters
[4].
Very
little
is
known
about
the
chemical
differences
between
these
lineages.
In
the
current
study,
we
have
used
both
genetic
sequencing
and
metabolic
profiling
to
characterize
samples
of
the
three
A.
taxiformis
lineages
found
in
Hawaiian
waters.
Sequencing
of
the
mitochondrial
cytochrome
oxidase
I
gene
clearly
separated
the
three
lineages.
In
parallel,
GC-‐MS
profiling
was
conducted
in
order
to
determine
whether
there
were
any
accompanying
chemical
differ-‐
ences.
Analysis
of
the
metabolite
profiles
using
a
range
of
multivariate
statistical
techniques
showed
that
one
of
the
three
lineages
could
be
differentiated
by
its
chemical
profile.
Several
halogenated
metabolites
characteristic
of
this
lineage
were
also
identified,
including
methyl
dibromoacetate,
methyl
tribromoacetate,
carbon
tetrabromide,
and
dibromoacetone.
Acknowledgements:
This
project
was
supported
in
part
by
the
NSF
Hawaii
EPSCoR
Program
under
NSF
Award
EPS-‐0903833
and
by
the
College
of
Pharmacy,
University
of
Hawaii
at
Hilo
Keywords: Chemotaxonomy, metabolomics, algae, Asparagopsis
References:
[1] Abbott
IA.
Limu:
An
ethnobotanical
study
of
some
Hawaiian
seaweeds.
National
Tropical
Botanical
Garden:
Lawai,
USA,
1984;
p
35
[2] Burreson
BJ,
Moore
RE,
Roller
PP.
Volatile
halogen
compounds
in
the
alga
Asparagopsis
taxiformis
(Rhodophyta).
J
Agric
Food
Chem
1976;
24:
856-‐861
[3] Andreakis
N,
Procaccini
G,
Kooistra
WHCF.
Asparagopsis
taxiformis
and
Asparagopsis
ar-‐
mata
(Bonnemaisoniales,
Rhodophyta):
genetic
and
morphological
identification
of
Med-‐
iterranean
populations.
Europ
J
Phycol.
2004,
39:
273-‐283
[4] Sherwood
AR.
Phylogeography
of
Asparagopsis
taxiformis
(Bonnemaisoniales,
Rhodo-‐
phyta)
in
the
Hawaiian
Islands:
Two
Mtdna
Markers
Support
Three
Separate
Introduc-‐
tions.
Phycologia
2008;
47:
79-‐88
P555
Antioxidant
and
antitumor
studies
of
Phyllophora
pseudocer-‐
anoides
extracts
Adriana
Trifan1,
Laura
Bucur2,
Daciana
Sava3,
Camelia
Paula
Stefanache4,
Ana
Clara
Aproto-‐
soaie1,
Oana
Cioanca1,
Monica
Hancianu1,
Anca
Miron1
1Department
of
Pharmacognosy,
University
of
Medicine
and
Pharmacy
”Grigore
T.
Popa”
Iași,
Faculty
of
Pharmacy,
16,
Universităţii
Street,700115,
Iași,
Romania,
2Department
of
Pharmacognosy,
”Ovidius”
Uni-‐
versity
Constanţa,
1,
Universităţii
Street,
Campus,
Bilding
B,
900470,
Constanţa,
Romania,
3Department
of
Natural
Sciences,
Faculty
of
Natural
and
Agricultural
Sciences,
”Ovidius”
University
Constanţa,
1,
Univer-‐
sităţii
Street,
Campus,
Bilding
B,
900470,
Constanţa,
Romania,
4
NIRDBS
/
“Stejarul”
Biological
Research
Centre,
Alexandru
cel
Bun
no.
6,
610004
Piatra
Neamt
Marine
macroalgae
have
attracted
an
emerging
interest
mainly
due
to
their
bioactive
constit-‐
uents
endowed
with
antioxidant,
antimicrobial,
anti-‐inflammatory
and
antitumor
properties
[1,
2].
The
aim
of
the
present
study
was
to
isolate,
characterize
and
assess
the
in
vitro
antioxi-‐
dant
and
antitumor
activities
of
the
crude
polysaccharide
and
phenolic
extracts
from
Phyllo-‐
phora
pseudoceranoides,
a
red
seaweed
commonly
found
in
the
Romanian
Black
Sea
coastal
waters.
The
water
soluble
polysaccharide
was
characterized
by
Fourier
transform
infrared
spectroscopy
(FT-‐IR).
Phenolic
compounds
were
extracted
using
solvents
of
different
polari-‐
ty:
methanol,
ethanol
and
water.
Total
phenolic
content
was
determined
by
Folin-‐Ciocalteu
method
[3].
The
antioxidant
activity
was
assessed
using
three
methods
DPPH
radical
scaveng-‐
ing
assay
[4],
ABTS
radical
cation
scavenging
assay
[5]
and
reducing
power
assay
[6].
Anti-‐
tumoural
activity
was
evaluated
on
human
cervical
adenocarcinomacell
line
(HeLacells),
us-‐
ingthe
MTT
assay
[7].
FT-‐IR
analysis
of
the
crude
polysaccharide
showed
characteristic
bands
of
carrageenan-‐type
structure.
Phenolic
extracts
displayed
moderate
DPPH
scavenging
activi-‐
ty,
with
values
ranging
from
12.99
±
0.23
to
46.00
±
0.13
at
2
mg/mL
;
all
extracts
showed
good
scavenging
activity
against
ABTS
radical,
with
IC50
values
ranging
from
0.12
±
0.02
to
0.49
±
0.02
mg/mL.
The
water
extract
exhibited
both
the
highest
phenolic
content
and
highest
antioxidant
activity.
At
1.5
mg/mL,
the
crude
polysaccharide
exhibited
important
ABTS
scav-‐
enging
activity
(49.59
±
0.03%)
and
showed
a
good
reducing
power
(0.406
±
0.002).
All
inves-‐
tigated
extractives
exhibited
strong
cytotoxic
activities
against
HeLacells.
The
antioxidant
and
antitumour
effects
of
the
sulfated
polysaccharide
and
phenolic
extracts
from
P.
pseudocer-‐
anoides
suggests
their
possible
use
as
ingredients
and
preservatives
in
the
food
and
pharma-‐
ceutical
industries,
but
also
as
a
source
of
potential
therapeutic
agents.
Acknowledgements:
This
paper
was
published
under
the
frame
of
European
Social
Found,
Human
Re-‐
sources
Development
Operational
Programme
2007-‐2013,
project
no.
POSDRU/159/1.5/136893.
Keywords: Phyllophora pseudoceranoides, algae, polysaccharides, phenolics, antioxidant ac-‐
tivity, antitumor activity.
References:
[1] Brown
EM,
Allsopp
PJ,
Magee
PJ,
Gill
CIR,
Nitecki
S,
Strain
CR,
McSorley
EM.
Seaweed
and
human
health.
Nutr
Rev
2014;
72:
205–216
[2] Michalak
I,
Chojnacka
K.
Algae
as
production
systems
of
bioactive
compounds.
Eng
Life
Sci
2015;
15:
160–176
[3] Wangensteen
H,
Samuelsen
AB,
Malterud
KE.
Antioxidant
activity
in
extracts
from
corian-‐
der.
Food
Chem
2004;
88:
293–297
[4] Malterud
KE,
Farbrot
TL,
Huse
AE,
Sund
RB.
Antioxidant
and
radical-‐scavenging
effects
of
anthraquinones
and
anthrones.
Pharmacology
1993;
47:
77–85
[5] Re
R,
Pellegrini
N,
Proteggente
A,
Pannala
A,
Yang
M,
Rice-‐Evans
C.
Antioxidant
activity
applying
an
improved
ABTS
radical
cation
decolorization
assay.
Free
Radic
Biol
Med
1999;
26:
1231−1237
[6] Berker
IK,
Güçlü
K,
Tor
I,
Apak
R.
Comparative
evaluation
of
Fe
(III)
reducing
power-‐based
antioxidant
capacity
assays
in
the
presence
of
phenanthroline,
batho-‐phenanthroline,
tri-‐
pyridyltriazine
(FRAP),
and
ferricyanide
reagents.
Talanta
2007;
72:
1157–1165
[7] Mosmann
T.
Rapid
colorimetric
assay
for
cellular
growth
and
survival:
application
to
pro-‐
liferation
and
cytotoxicity
assays.
J
Immunol
Methods
1983;
65:
55-‐63.
P556
Preparative
separation
of
marine
bioactive
compounds
by
cen-‐
trifugal
partition
chromatography
Romain
Boulho1,
Julie
Le
Roux2,
Céline
Le
Quemener2,
Anne-‐Sophie
Burlot1,
Grégoire
Audo2,
Nathalie
Bourgougnon1,
Gilles
Bedoux1
1
Univ.
Bretagne-‐Sud,
EA
3884,
LBCM,
IUEM,
F-‐56000
Vannes,
France.
2
ARMEN
INSTRUMENT,
16,
Rue
Ampère, 56890 Saint-‐Avé.
In
response
to
stresses,
the
seaweeds
are
known
to
possess
different
chemical
defense
mech-‐
anisms
by
secondary
metabolites
production.
The
aim
of
this
study
is
to
present
the
Centrifu-‐
gal
Partition
Chromatography
(CPC)
for
the
fractionation
of
bioactive
molecules
isolated
from
the
red
seaweed
Solieria
chordalis
(Gigartinales,
Solieriaceae).
This
separative
technique
is
known
for
its
low
solvent
consumption.
CPC
was
first
applied
for
the
separation
of
crude
samples
prepared
in
various
solvents.
The
determination
of
the
mobile
phase
was
based
on
the
partition
of
metabolites
between
two
polar
and
apolar
solvent
systems
which
belong
to
the
Arizona
solvent
system
[1].
Anti-‐UVB
capacity
and
anti-‐aging
activity
were
studied
and
used
for
the
bioassay-‐guided
fractionation.
The
crude
extracts
were
prepared
with
yields
ranging
from
1.0
%
(dw/dw)
(ethyl
acetate)
to
19.6
%
(dw/dw)
(methanol).
Thin
layer
chromatography
analysis
led
to
the
determination
of
the
solvent
biphasic
system
composition.
The
mode
of
elution
and
the
mobile
phase
composi-‐
tion
influenced
the
nature
and
the
number
of
collected
fractions.
So
far,
this
technique
has
allowed
separating
concentrated
fractions
characterized
by
their
UV-‐spectrum
and
LC-‐MS
profile.
Among
the
different
compounds,
the
UV-‐absorbing
Mycosporine-‐like
Amino
Acid
(MAA)
palythenic
acid
was
detected
[2].
The
preliminary
anti-‐UVB
test
has
shown
a
positive
photoprotective
activity
by
an
important
increase
of
the
half-‐life
t1/2
of
chlorophyll.
Moreo-‐
ver,
additional
anti-‐elastase
activity
was
shown
for
different
fractions
[3].
In
conclusion,
these
results
highlighted
the
hypothesis
that
the
anti-‐UVB
activity
of
the
extract
is
correlated
to
the
presence
of
the
MAAs.
Tests
are
in
progress
in
order
to
evaluate
the
frac-‐
tions
obtained
by
CPC
in
order
to
characterize
the
structure
and
the
bioactivity
levels
of
the
bioactive
compounds.
Acknowledgements: Financial supports: The Region Bretagne and The Department of Morbihan.
Keywords: Soliera chordalis, bioactive molecules, centrifugal partition chromatography, frac-‐
tionation
References:
[1] Berthod
A
,
Hassoun
M,
Harris
G.
Using
the
Liquid
Nature
of
the
Stationary
Phase:
The
Elution-‐Extrusion
Method.
J
Liq
Chromatogr
Relat
Technol
2005;
28:
1851–1866
[2] Bedoux
G,
Hardouin
K,
Marty
C,
Taupin
L,
Vandanjon
L,
Bourgougnon
N.
Chemical
characterization
and
photoprotective
activity
measurement
of
extracts
from
the
red
macroalga
Solieria
chordalis.
Bot
Mar
2014;
57:
291–301
[3] Bedoux
G,
Hardouin
K,
Burlot
AS,
Bourgougnon
N.
Bioactive
components
from
seaweeds:
Cosmetic
applications
and
future
development.
Adv
Bot
Res
2014;
71:
345-‐378
P557
Caulerpenyne
from
Caulerpa
Taxifolia:
a
comparative
study
be-‐
tween
CPC
and
classical
chromatographic
techniques
Estelle
Sfecci1,
Céline
le
Quémener2,
Grégoire
Audo2,
Thierry
Lacour3,
Philippe
Amade1,
Mo-‐
hamed
Mehiri1
1
Institut
de
Chimie
de
Nice,
UMR-‐CNRS
7272,
Marine
Natural
Products
Team,
Faculté
des
Science,
Uni-‐
versité
Nice
Sophia
Antipolis,
Parc
Valrose,
06108
Nice
Cedex
02,
France,
2
ARMEN
Instrument,
ZI
Kermel-‐
in,
16
rue
Ampère,
56890
Saint-‐Avé,
France,
3
BioPreserv,
4
traverse
Dupont,
Grasse,
France.
Caulerpenyne
(Cyn)
is
a
cytotoxic
compound
firstly
isolated
in
1978
from
Caulerpa
prolifera
[1].
This
molecule,
constituted
by
a
diacetoxybutadiene
moiety,
exhibited
a
wide
range
of
bio-‐
logical
properties
with
mainly
antibacterial
properties
[2]
and
antitumoral
activities
by
inhib-‐
iting
the
growth
of
several
human
cancer
cell
lines
[3].
Several
industrial
applications
are
pos-‐
sible
for
Cyn,
so
there
is
a
need
to
produce
and
isolate
it
in
large
quantities.
Since
Cyn
purifica-‐
tion
is
time-‐
and
solvent-‐consuming,
it
is
crucial
to
find
a
more
green
process
to
obtain
pure
Cyn
with
lower
costs.
Thus,
in
our
study,
Cyn
has
been
purified
from
C.
taxifolia
crude
extract
with
two
different
techniques:
Centrifugal
Partition
Chromatography
(CPC)
and
classical
chromatographic
techniques.
CPC
method
involves
only
the
CPC
step
with
0.2%
yield
(dry
weight).
On
the
other
hand,
other
chromatographic
techniques
traditionally
used
imply
at
least
three
steps:
(i)
a
liquid-‐liquid
extraction,
(ii)
a
size
exclusion
chromatography,
and
final-‐
ly
(iii)
a
diol
column
chromatography
with
a
0.04%
yield
(dry
weight).
Among
the
current
chromatographic
techniques,
CPC
seemed
the
more
appropriate
to
our
objectives
for
several
reasons:
(i)
it
allows
low
solvent
consumption,
(ii)
it
can
be
used
from
analytical
to
prepara-‐
tive
scale
and
(iii)
it
is
less
time-‐consuming
than
other
techniques.
In
the
literature,
CPC
has
been
used
to
fractionate
and/or
isolate
bioactive
compounds
and
has
shown
its
efficiency
in
the
purification
process
of
natural
products
from
diverse
species/
origin.
The
comparative
study
showed
CPC
to
be
faster
at
lower
costs
for
Cyn
isolation,
and
increased
the
extraction
yield
significantly.
Acknowledgements:
E.
Sfecci
is
the
recipient
of
a
thesis
grant
from
the
"Conseil
Régional
Provence
Alpes
Côte
d’azur".
M.
Mehiri
research
is
supported
by
the
french
program
ENVI-‐Med
"MEDIBIO",
the
ANR/Investissements
d’Avenir
program
via
the
OCEANOMICs
project
(grant
#ANR-‐11-‐BTBR-‐0008),
and
the
H2020
European
program
via
the
EMBRIC
project.
Keywords: Caulerpa taxifolia, caulerpenyne, centrifugal partition chromatography, isolation,
purification.
References:
[1] Amico
V,
Oriente
G,
Piattelli
M,
Tringali
C,
Fattorusso
E,
Magno
S,
Mayol
L.
Caulerpenyne,
an
unusual
sequiterpenoid
from
the
green
alga
Caulerpa
prolifera.
Tetrahedron
Lett
1978;
19:
3593
–
3596
[2] Hodgson
LM.
Antimicrobial
and
antineoplastic
activity
in
some
south
Florida
seaweeds.
Bot
Mar
1984;
27:
387
–
390
[3] Fischel
JL,
Lemée
R,
Formento
P,
Caldani
C,
Moll
JL,
Pesando
D,
Meinesz
A,
Grelier
P,
Pietra
F,
Guerriero
A,
Milano
G.
Cell
growth
inhibitory
effects
of
caulerpenyne,
a
sesquit-‐
erpenoid
from
the
marine
alga
Caulerpa
taxifolia.
Anticancer
Res
1995;
15:
2155
–
2160
P558
Study
on
the
antibacterial
components
from
Palythoa-‐symbiotic
fungus,
Fusarium
solani
Chih-‐Chuang
Liaw,
Po-‐Yi
Cheng
Department
of
Marine
Biotechnology
and
Resources,
National
Sun
Yat-‐sen
University,
70
Lienhai
Rd.,
Kaohsiung
80424,
Taiwan
Symbiotic
microorganisms
have
been
found
to
interfere
the
host’s
metabolism
system
to
pro-‐
duce
toxins
or
anti-‐feedants/
or
directly
produce
these
metabolites
to
store
in
the
organs
of
the
hosts
to
protect
their
host
from
predators
and
grazers
[1].
In
the
view
of
natural
products
chemistry,
these
bioactive
compounds
have
long
been
a
central
to
drug
discovery
and
organic
chemistry
[2].
Based
on
the
preliminary
anti-‐microbial
activity
screening,
we
found
some
ma-‐
rine
symbiotic
microbes,
which
we
isolated
from
the
marine
invertebrates
collected
from
the
intertidal
zones
in
Taiwan,
showed
clear
inhibitory
effects
on
certain
pathogens.
We
are
in-‐
terested
in
exploring
the
bioactive
compounds
from
these
marine
symbiotic
microbes.
A
sym-‐
biotic
fungus
FS-‐01
was
isolated
from
a
Palythoa
sp.,
which
was
collected
from
Wanlitong,
Pingtung
County,
Taiwan.
The
internal
transcribed
spacer
(ITS)
sequencing
analysis
identified
the
fungal
strain
FS-‐01
as
Fusarium
solani.
Interestingly,
the
different
cultural
condition
to
FS-‐
01
made
the
generation
of
different
colorful
metabolites.
The
reddish
acetyl
acetate
extract
of
FS-‐01
cultured
in
the
thick
medium
showed
inhibitory
effects
against
Acinetobacter
bau-‐
mannii
and
Bacillus
cereus,
while
the
white
one
cultured
in
the
thin
medium
only
showed
the
inhibitory
effect
against
B.
cereus.
Based
on
the
antibacterial
activity-‐directed
fractionation
and
HPLC
isolation,
we
isolated
a
series
of
reddish
compounds
(FS01
and
FS03-‐09)
and
one
polyketide
FS02
from
the
EA
extract
of
FS-‐01
cultured
in
thick
medium.
All
the
isolates
were
elucidated
by
the
Mass
and
1D
and
2D
NMR
spectral
data,
whereas
known
ones
were
identi-‐
fied
by
comparison
with
literature
data.
Among
them,
compounds
FS01
and
FS02
are
one
new
1,
4-‐naphthoquinone
as
5-‐hydroxy-‐8-‐methoxy-‐2,
4-‐dimethyl-‐1H-‐benzo[g]indole-‐6,
9-‐
dione
and
one
new
polyketide
derivative
as
10-‐hydroxy-‐3,
5,
7-‐trimethylundec-‐6-‐ene-‐2,
8-‐
dione,
respectively.
Furthermore,
the
anti-‐microbial
activity
of
these
isolates
is
under
investi-‐
gation.
Key
word:
Palythoa
sp.,
symbiotic
fungus,
Fusarium
solani,
1,4-‐naphthoquinone,
polyketide
References:
[1] Hay
ME.
Marine
chemical
ecology:
Chemical
signals
and
cues
structure
marine
popula-‐
tions,
communities,
and
ecosystems.
Ann
Rev
Mar
Sci
2009;
1:
193-‐212
[2] Imhoff
JF,
Labes
A,
Wiese
J.
Bio-‐mining
the
microbial
treasures
of
the
ocean:
New
natural
products.
Biotechnol
Adv
2011;
29:
468-‐482
P559
Metabolite
profiling
of
Baltic
blue
mussel
(Mytilus
sp.)
hybrids
and
their
associated
microbes
Caroline
Utermann1,
Corinna
Breusing2,
Heiko
Stuckas3,
Tim
Staufenberger4,
Antje
Labes1,
Deniz
Tasdemir1
1
Research
Unit
Marine
Natural
Products
Chemistry,
GEOMAR
Centre
for
Marine
Biotechnology
(GE-‐
OMAR-‐Biotech),
GEOMAR
Helmholtz
Centre
for
Ocean
Research
Kiel,
Am
Kiel-‐Kanal
44,
24106
Kiel,
Ger-‐
many,
2
Research
Unit
Evolutionary
Ecology
of
Marine
Fishes,
GEOMAR
Helmholtz
Centre
for
Ocean
Re-‐
search
Kiel,
Düsternbrooker
Weg
20,
24105
Kiel,
Germany,
3
Senckenberg
Natural
History
Collection
Dresden,
Population
Genetics,
Koenigsbruecker
Landstrasse
159,
01109
Dresden,
Germany,
4
Kieler
Meeresfarm
UG,
Richthofenstrasse
31,
24159
Kiel,
Germany
The
blue
mussel
genus
Mytilus
is
a
popular
food
source
and
an
important
model
organism
for
many
scientific
disciplines
[1,2].
The
morphologically
similar
species
M.
edulis
(Me)
and
M.
trossulus
(Mt)
form
an
unprecedented
and
unique
hybridisation
pattern
in
the
Baltic
Sea
[3,
4].
Recently,
it
was
shown
that
metabolite
profiling
of
Mytilus
species
provides
significant
po-‐
tential
for
biomonitoring
of
aquatic
environments
but
also
for
informing
natural
product
re-‐
search
[1,2].
However,
the
metabolome
of
Mytilus
sp.
is
still
little
investigated
and
no
metabo-‐
lome
data
from
the
Baltic
Sea
blue
mussels
are
available
so
far.
In
order
to
compare
the
me-‐
tabolite
profiles
of
Baltic
Mytilus
sp.
at
an
individual
level,
30
Mytilus
specimens
were
sampled
at
a
blue
mussel
farm
in
Kiel
(Baltic
Sea)
and
genotyped
with
two
commonly
utilized
nuclear
markers,
EFbis
and
Glu-‐5'
[4].
Genotypic
assessment
revealed
a
dominance
of
Mt
alleles
at
Kiel
Fjord
with
90%
of
the
samples
being
intermediate
(Me/Mt)
or
Mt-‐like
hybrids.
Secondary
metabolites
of
the
whole
tissue
extracts
(EtOAc)
were
explored
by
HPLC-‐DAD-‐MS.
Metabolite
profiles
were
found
to
be
similar
but
not
identical,
differing
mainly
in
minor
nonpolar
second-‐
ary
metabolites.
Considering
that
secondary
metabolites
are
often
produced
by
associated
microbes
and
not
by
the
macroorganism
itself,
the
microbiome
of
the
mussel
specimens
was
also
investigated.
A
culture-‐based
approach
yielded
259
bacterial
and
fungal
strains,
including
Actinomycetes.
Bacteria
belonging
to
the
orders
Alteromonadales
and
Pseudomonadales
(Acinetobacter
sp.,
Pseudomonas
sp.,
Pseudoalteromonas
sp.,
Psychrobacter
sp.,
Shewanella
sp.)
dominated
the
microbial
community
(57
%).
In
addition,
an
RFLP
based
culture-‐independent
approach
was
developed
for
Mytilus
sp.
individuals.
Both
the
culture-‐dependent
and
culture-‐
independent
approaches
showed
inter-‐individual
differences
with
respect
to
the
associated
microbes,
but
no
genotypic
dependence.
Acknowledgements:
Prof.
Dr.
Thorsten
Reusch
(GEOMAR)
is
acknowledged
for
providing
access
to
his
facilities
for
genotyping
studies
Keywords: Blue mussel, Mytilus, secondary metabolite profiling, HPLC-‐DAD-‐MS, microbiome,
RFLP
References:
[1] Bundy
JG,
Davey
MP,
Viant
MR.
Environmental
metabolomics:
a
critical
review
and
future
perspectives.
Metabolomics
2009;
5:
3-‐21
[2] Grienke
U,
Silke
J,
Tasdemir
D.
Bioactive
compounds
from
marine
mussels
and
their
effects
on
human
health.
Food
Chem
2014;
142:
48-‐60
[3] Stuckas
H,
Stoof
K,
Quesada
H,
Tiedemann
R.
Evolutionary
implications
of
discordant
clines
across
the
Baltic
Mytilus
hybrid
zone
(Mytilus
edulis
and
Mytilus
trossulus).
Heredity
2009;
103:
146-‐156
[4] Breusing
C.
Population
genetics
and
morphometric
variation
of
blue
mussels
in
the
west-‐
ern
Baltic
Sea.
Master
thesis
2012,
Christian-‐Albrechts-‐Universität
Kiel,
Germany
P560
Chemical
and
biological
screening
of
deep-‐water
sponges
from
Antarctic
regions
Fengie
Li1,
Michael
Marner1,
Arlette
Wenzel-‐Storjohann1,
Johanna
Silber1,
Dorte
Janussen2,
Deniz
Tasdemir1
1
Research
Unit
Marine
Natural
Products
Chemistry,
GEOMAR
Centre
for
Marine
Biotechnology
(GE-‐
OMAR-‐Biotech),
GEOMAR
Helmholtz
Centre
for
Ocean
Research
Kiel,
Am
Kiel-‐Kanal
44,
24106
Kiel,
Ger-‐
many,
2
Senckenberg
Research
Institute
and
Natural
History
Museum,
Senckenberganlage
25,
60325
Frankfurt,
Germany
The
majority
of
marine
natural
products
(MNPs)
originate
from
tropical
and
temperate
shal-‐
low
water
invertebrates,
such
as
sponges
[1].
Recent
studies
indicate
the
presence
of
great
genetic
diversity
in
deep-‐waters
that
may
be
linked
to
unprecedented
chemistry
due
to
evolu-‐
tion/adaptation
to
extremely
harsh
environmental
conditions.
However,
only
less
than
2%
of
MNPs
derive
from
the
deep-‐sea
organisms
[2].
Antarctic
ecosystems
are
rich
in
biodiversity
[3]
and
exposed
to
unique
environmental
characteristics
resulting
in
communities
structured
both
by
biotic
interactions
(e.g.
predation,
competition)
and
abiotic
factors
(e.g.
seasonality,
ice-‐scouring)
[4],
suggesting
a
high
chemical
diversity.
In
this
work,
we
investigated
39
deep-‐
water
sponges
collected
from
the
Antarctic
Weddell
Sea
and
adjacent
areas
(depths
-‐100-‐
600m).
The
freeze-‐dried
sponge
samples
were
extracted
with
water,
followed
by
MeOH
and
CH2Cl2
separately.
The
combined
organic
extracts
were
tested
for
activity
against
cancer
cells
[HepG2
(liver)
and
HT29
(bowel)
cancer
cell
lines],
bacteria
[ESKAPE
panel:
Enterococcus
fae-‐
calis,
Staphylococcus
aureus
(MRSA),
Klebsiella
pneumoniae,
Acinetobacter
baumannii,
Pseu-‐
domonas
aeruginosa,
Escherichia
coli)
and
fungi
(yeasts
Candida
albicans,
Cryptococcus
neoformans).
Several
Latrunculia
sponge
extracts
displayed
high
anticancer
activity
against
both
cell
lines
(IC50
values
0.50-‐3.16
µg/ml).
The
organic
extract
of
the
glass
sponge
Rossella
cf.
antarctica
showed
moderate
antibiotic
activity
towards
MRSA
and
E.
faecalis
with
IC50
val-‐
ues
of
96
and
213
µg/ml,
respectively.
All
extracts
have
undergone
chemical
profil-‐
ing/dereplication
studies
by
HPLC-‐DAD-‐MS
and
1H
NMR
spectroscopy.
The
results
of
chemi-‐
cal
and
biological
screening
will
assist
in
selection
and
activity-‐guided
isolation
of
Antarctic
deep-‐water
sponge
metabolites.
Acknowledgements:
China
Scholarship
Council
is
acknowledged
for
funding
Keywords: Antarctic deep-‐water sponge, screening, anticancer, antibacterial, dereplication
References:
[1] Leal
MC,
Puga
J,
Serôdio
J,
Gomes
NC,
Calado
R.
Trends
in
the
discovery
of
new
marine
nat-‐
ural
products
from
invertebrates
over
the
last
two
decades
-‐
where
and
what
are
we
bio-‐
prospecting?
PLoS
One
2012;
7:
e30580
[2] Skropeta
D,
Wei
LQ.
Recent
advances
in
deep-‐sea
natural
products.
Nat
Prod
Rep
2014;
31:
999-‐1025
[3] Brandt
A,
Gooday
AJ,
Brandão
SN,
Brix
S,
Brökeland
W,
Cedhagen
T,
Choudhury
M,
Cor-‐
nelius
N,
Danis
B,
De
Mesel
I,
Diaz
RJ,
Gillan
DC,
Ebbe
B,
Howe
JA,
Janussen
D,
Kaiser
S,
Linse
K,
Malyutina
M,
Pawlowski
J,
Raupach
M,
Vanreusel
A.
First
insights
into
the
biodi-‐
versity
and
biogeography
of
the
Southern
Ocean
deep
sea.
Nature
2007;
447:
307-‐311
[4] Avila
C,
Taboada
S,
Núñez-‐Pons
L.
Antarctic
marine
chemical
ecology:
what
is
next?
Mar
Ecol
2008;
29:
1-‐71
P561
Characterization
of
bioactive
molecules
derived
from
marine
mi-‐
croorganisms,
interfering
with
cell
dedifferentiations
observed
at
the
invasive
front
of
mammary
tumors
Ambre
Dezaire1,
2,
Nathalie
Ferrand1,
Marine
Vallet2,
Michèle
Sabbah1,
Annette
K.
Larsen1,
Soizic
Prado2,
Alexandre
Escargueil1
1
INSERM
UMR_S
938
Hôpital
Saint
Antoine
Bâtiment
Kourilsky
184,
rue
du
Faubourg
St
Antoine
75571
Paris
Cedex
12,
2
UMR
7245,
MCAM,
CNRS,
Muséum
National
d’Histoire
Naturelle,
63
Rue
Buffon,
75005
Paris,
France
Breast
cancer
is
the
leading
cause
of
cancer
death
in
women.
At
the
in
situ
carcinoma
stage
the
tumor
can
be
surgically
removed,
but
in
later
stages
tumor
cells
can
undergo
epithelial-‐to-‐
mesenchymal
transition
(EMT)
[1],
become
invasive
and
chemoresistant
[2].
To
date,
re-‐
search
on
therapy
targeting
EMT
[3]
does
not
consider
cellular
heterogeneity
of
the
tumor.
Indeed,
tumor
cells
communicate
particularly
with
adipocytes
[4],
resulting
in
an
EMT,
a
loss
of
estrogen
receptors
and
adipocyte
dedifferentiation.
Recent
examples
of
marine
products
interfering
with
these
dedifferentiation
processes
have
been
highlighted
[5].
In
addition,
ma-‐
rine
biodiversity,
from
which
microorganisms
[6],
already
led
to
several
drugs
on
the
market
[7],
especially
in
oncotherapy.
Our
experimental
strategy
is
to
isolate
and
characterize
molecules
extracted
from
70
fungal
strains
associated
to
brown
algae
and
cultivated
in
two
conditions.
Extracts
and
molecules
are
selected
on
their
ability
to
inhibit
tumor
cell
invasion,
and
adipocyte-‐tumor
cell
communica-‐
tion
by
measuring
both
cell
line
differentiation
states.
A
first
viability
assay
has
shown
that
16
strains
have
high
cytotoxic
effects
(IC50
<
25
µg
/
mL).
A
strain
from
the
marine
fungus
Pa-‐
radendryphiella
arenaria
was
particularly
efficient
with
an
IC50
of
1
µg
/
mL
on
MCF7
epithe-‐
lial
cancer
cell
lines
and
0.7
µg
/
mL
on
MCF7-‐Sh-‐WISP2
invasive
cancer
cell
lines.
The
chem-‐
istry
of
this
fungus
is
poorly
studied.
An
ongoing
purification
and
dereplication
step,
will
al-‐
low
for
isolation
of
potentially
new
bioactive
molecules.
These
experiments
let
us
bring
out,
yet,
non
studied
marine
fungal
crude
extracts
that
have
an
anticancer
activity,
not
only
on
non-‐invasive
mammary
cancer
cells,
but
also
on
invasive
ones,
which
constitute
their
strong
therapeutic
potential.
Acknowledgements:
The
UPMC
through
the
PDIF
and
Lapervenchelif
are
acknowledged
for
their
finan-‐
cial
support
Keywords:
Breast
cancer,
EMT,
marine
natural
products,
fungal
endophytes,
Paradendryphi-‐
ella
arenaria
References:
[1] Radisky
DC.
Epithelial-‐mesenchymal
transition.
J
Cell
Sci
2005;
118:4325-‐4326
[2] Thomson
S,
Buck
E,
Petti
F,
Griffin
G,
Brown
E,
Ramnarine
N,
Iwata
KK,
Gibson
N,
Haley
JD.
Epithelial
to
mesenchymal
transition
is
a
determinant
of
sensitivity
of
non–small-‐cell
lung
carcinoma
cell
lines
and
xenografts
to
epidermal
growth
factor
receptor
inhibition.
Can-‐
cer
Res
2005;
65:
9455-‐62
[3] Chua
KN,
Sim
WJ,
Racine
V,
Lee
SY,
Goth
BC,
Thiery
JP.
A
cell-‐Based
small
molecule
screen-‐
ing
method
for
identifying
inhibitors
of
epithelial-‐
mesenchymal
transition
in
carcinoma.
PLoS
One
2012;
7:e33183
[4] Mueller
MM,
Fusenig
NE.
Friends
or
foes
-‐
bipolar
effects
of
the
tumor
stroma
in
cancer.
Nat
Rev
Cancer
2004;
4:839-‐49
[5] Charytonowicz
E,
Terry
M,
Coakley
K,
Telis
L,
Remotti
F,
Cordon-‐Cardo
C,
Taub
RN,
Matu-‐
shansky
I.
PPARγ
agonists
enhance
ET-‐743–induced
adipogenic
differentiation
in
a
trans-‐
genic
mouse
model
of
myxoid
round
cell
liposarcoma.
J
Clin
Invest
2012;
22:886-‐98.
[6] Debbab
A,
Aly
AH,
Proksch
P.
Endophytes
and
associated
marine
derived
fungi
-‐
ecological
and
chemical
perspectives.
Fungal
Div
2012;
57:45-‐83
[7] Gerwick
WH,
Fenner
AM.
Drug
discovery
from
marine
microbes.
Microb
Ecol
2013;
65:800-‐806
[8] Ferrand
N,
Gnanapragasam
A,
Dorothee
G,
Redeuilh
G,
Larsen
AK,
Sabbah
M.
Loss
of
WISP2/CCN5
in
estrogen-‐dependent
MCF7
human
breast
cancer
cells
promotes
a
stem-‐
like
cell
phyenotype.
PLoS
One
2014;
9:e87878
P562
Botryane
sesquiterpenes
and
binaphthalene
tetrols
from
endo-‐
phytic
fungi
associated
to
the
marine
red
algae
Asparagopsis
taxi-‐
formis
Rebeca
P.
Medina1,
Airton
D.
Silva1,
Raymond
J.
Andersen2,
Angela
R.
Araújo,
Dulce
H.
S.
Silva1
1
Institute
of
Chemistry,
University
of
São
Paulo
State
-‐
Street
Professor
Francisco
Degni,
55,
14800-‐060,
Araraquara,
Brazil,
2
Department
of
Chemistry,
University
of
British
Columbia,
2036
Main
Mall,
Vancou-‐
ver,
British
Columbia
V6T
1Z1,
Canada
Marine
organisms
may
host
a
variety
of
endophytic
fungi,
which
induce
or
enhance
produc-‐
tion
of
special
metabolites
associated
to
important
features
in
adaptation,
defence
against
predators,
and
represent
important
sources
for
bioprospection.
Asparagopsis
taxiformis
is
a
marine
red
algae
spread
throughout
Brazilian
coast.
Specimens
from
its
Falkenbergia
stage
were
collected
at
the
rocky
shore
of
Fortaleza
Beach,
Ubatuba,
SP,
Brazil
and
surface
steri-‐
lized,
fragmented
and
spread
on
PDA
plates,
which
led
to
the
isolation
of
seven
fungal
strains
including
Nemania
bipapillata
and
Hypoxylon
investiens.
After
cultivation
of
each
strain
in
ster-‐
ilized
PDB
medium
with
ultrapure
water
or
sea
water
at
250C,
each
broth
was
separated
from
the
mycelium
by
filtration
and
afforded
crude
extracts
by
extraction
with
EtOAc.
Fractionation
of
the
Nemania
bipapillata
extract
by
RP-‐CC
and
HPLC
afforded
4-‐acetoxy-‐9,10,15-‐
trihydroxyprobotridial,
a
presilphiperfolane-‐type
sesquiterpene,
in
addition
to
two
novel
botryane
derivatives
related
to
those
isolated
from
Botrytis
cinerea
[1−3].
Their
molecular
formulas
were
determined
as
C14H22O3
and
C15H24O4
by
HRESI-‐TOF-‐MS,
which
confirmed
their
structures
after
1D
and
2D
NMR
spectral
analyses.
Metabolites
with
a
botryane
skeleton
have
shown
phytotoxic,
antibiotic
and
cytotoxic
activity
and
were
previously
isolated
from
fungi
associated
to
the
marine
red
algae
Polysiphonia
[4].
Hypoxylon
investiens
extract
was
chromatographed
over
Sephadex
LH-‐20,
purified
by
HPLC
and
afforded
two
novel
tetrols
which
had
their
structures
established
by
NMR
and
MS
analyses
as
BNT
(4,5,4',5'-‐
tetrahydroxy-‐1,1'-‐binaphthyl)
derivatives.
Such
compounds
are
structurally
related
to
Hy-‐
poxylonols
A-‐F
and
Daldenones
C-‐D,
previously
isolated
from
Xylariaceae
[5,
6].
Such
promis-‐
ing
results
highlight
the
outstanding
chemodiversity
of
marine-‐derived
fungi,
which
may
con-‐
tribute
for
the
sustainable
exploration
of
Brazilian
marine
biodiversity
in
the
search
for
bioac-‐
tive
compounds.
Acknowledgements: CAPES and FAPESP
Keywords: Nemania bipapillata, Hypoxylon investiens, endophytic fungus, sesquiterpene,
binaphthalene
References:
[1] Durán-‐Patrón
R,
Hernández-‐Galan
R,
Rebordinos
LG,
Cantoral
JM,
Collado
IG.
Structure-‐
activity
relationships
of
new
phytotoxic
metabolites
with
the
botryane
skeleton
from
Bo-‐
trytis
cinerea.
Tetrahedron
1999;
55:
2389-‐2400
[2] Collado
IG,
Hernández-‐Galan
R,
Durán-‐Patrón
R,
Cantoral
JM.
Metabolites
from
a
shake
culture
of
Botrytis
cinerea.
Phytochemstry
1995;
38:
647-‐650
[3] Durán-‐Patrón
R,
Hernández-‐Galan
R,
Collado
IG.
Secobotrytriendiol
and
related
sesquit-‐
erpenoids:
new
phytotoxic
metabolites
from
Botrytis
cinerea.
J
Nat
Prod
2000;
63:
182-‐
184
[4] Krohn
K,
Dai
J,
Flörke
U,
Aust
HJ,
Dräger
S,
Schulz
B.
Botryane
metabolites
from
the
fungus
Geniculosporium
sp.
isolated
from
the
marine
red
alga
Polysiphonia.
J
Nat
Prod
2005;
68:
400-‐405
[5] Fukai
M,
Suzuki
T,
Nagasawa
I,
Kinoshita
K,
Takahashi
K,
Koyama
K.
Antiangiogenic
activi-‐
ty
of
Hypoxylonol
C.
J
Nat
Prod
2014;
77:
1065-‐1068
[6] Quang
DN,
Hashimoto
T,
Nomura
Y,
Wollweber
H,
Hellwig
V,
Fournier
J,
Stadler
S,
Asakawa
Y.
Cohaerins
A
and
B,
azaphilones
from
the
fungus
Hypoxylon
coharens,
and
comparison
of
HPLC-‐based
metabolite
profiles
in
Hypoxylon
sect.
Annulata.
Phytochemistry
2005;
66:
797-‐809
P563
Cytotoxic
compounds
of
sponges
collected
from
West
Bali
Na-‐
tional
Park
Indonesia
Erna
Prawita
Setyowati,
Anggara
Jenie,
Sudarsono,
Subagus
Wahyuono
Department
of
Pharmaceutical
Biology,
Faculty
of
Pharmacy,
Gadjah
Mada
University,
Yogyakarta,
Indo-‐
nesia.
Investigations
of
the
structure
of
cytotoxic
compound
of
sponges
collected
from
West
Bali
National
Park
Indonesia
has
been
conducted.
The
structures
were
identified
using
ultraviolet
spectroscopy,
MS,
1H-‐NMR
and
13C-‐NMR.
Based
on
the
spectroscopic
data
and
comparison
with
literature,
the
compound
from
Stylissa
flabelliformis
was
identified
as
jaspamide
(1).
Compounds
from
Kaliapsis
sp
were
identified
as
theonellapeptolide
1d
(2)
and
1-‐(tetrahydro-‐
4-‐hydroxy-‐5-‐(hydroxymethyl)furan-‐2-‐yl)-‐5-‐methyl
pyrimidine-‐2,4(1H,3H)-‐dione
(3).
Cytotoxic
test
of
these
three
compounds
showed
that
(1)
has
LC50
=0.08
µg/ml
on
myeloma
cell.
The
IC50
of
(2)
was
10.3,
8.3,
16.5
and
7.8
µg/ml
on
myeloma,
T47D,
HeLa
and
Raji
cells
respectively.
The
IC50
of
(3)
was
0.18,
7.9,
6.9
and
5.8
µg/ml
on
myeloma,
T47D,
HeLa
and
Raji
cells
respectively.
O 60
56
58 N 53 55
64 63 N 59
H
10
52 57 8
O O 7
O NH O 14
1 3 O 12
6
51
O O O HN O
49 17
O
47
44 NH
46
HN
43
48 O
O N 25 HN O
H H O N H
40 N 20
39 37 N 34 N 29 23
24
28 N
21
H O
30
O O O
31
HO
OH
(1)
(2)
(3)
Keywords: Stylissa flabelliformis, jaspamide, Kaliapsis sp, theonellapeptolide 1d, 1-‐
(tetrahydro-‐4-‐hydroxy-‐5-‐(hydroxymethyl)furan-‐2-‐yl)-‐5-‐methyl pyrimidine-‐2,4(1H,3H)-‐dione
References:
[1] Setyowati
EP,
Wahyuono
SS.
Jaspamide:
Identifikasi
struktur
senyawa
sitotoksik
dan
fun-‐
gisid
dari
spons
Stylissa
flabelliformis.
Majalah
Farmasi
Indonesia
2015;
16:
12-‐19
[2] Setyowati
EP,
Jenie
UA,
Sudarsono,
Kardono
LB,
Rahmat
R.
Identification
of
cytotoxic
constituent
of
Indonesian
sponge
Kaliapsis
sp.
(Bowerbank).
Pak
J
Biol
Sci
2008;
11:
2560-‐2566
[3] Setyowati
EP,
Jenie
UA,
Sudarsono,
Kardono
LB.
Apoptosis
induction
of
bioactive
substance
Theonellapep
tolide
1d
and
1-‐(tetrahydro-‐4-‐hydroxy-‐5-‐(hydroxymethyl)furan-‐2-‐yl)-‐5-‐
methyl
pyrimidine-‐2,4(1H,3H)-‐dione
isolated
from
Kaliapsis
sp
sponge
collection
from
West
Bali
National
Park
Indonesia.
J
Biol
Sci
2016;
16:
30-‐36
P564
P-‐Sulfooxyphenylpyruvic acid derivatives from the green macroalga
Caulerpa taxifolia
Estelle Sfecci1, Lionel Massi1, Thierry Lacour2, Philippe Amade1, Mohamed Mehiri1.
1 Chemistry Institute of Nice, Marine Natural Products Team, Nice Sophia Antipolis University, UMR-‐CNRS
7272, Parc Valrose, 06108 Nice Cedex 02, France, 2 BioPreserv, 4 traverse Dupont, Grasse, France.
The
green
macroalga
Caulerpa
taxifolia
has
been
of
a
great
research
interest
since
its
acci-‐
dental
introduction
in
the
Mediterranean
sea
in
1984.
This
specie
has
yielded
several
struc-‐
turally
diverse
and
biologically
active
compounds
such
as
caulerpenyne
(Fig.
1)
[1].
This
me-‐
tabolite,
constituted
by
a
diacetoxybutadiene
moiety,
exhibited
very
potent
antibacterial
properties
[2].
Within
our
ongoing
research
projects
devoted
to
the
isolation
of
new
bioactive
metabolites
from
Mediterranean
marine
organisms,
a
specimen
of
the
freshly
collected
green
algae
Caulerpa
taxifolia,
was
investigated.
The
crude
polar
organic
extract
was
subjected
to
a
series
of
chromatographic
separations
that
has
led
to
the
isolation
of
new
metabolites
(1a
and
1b)
structurally
close
to
the
already
reported
ones
(2a
and
2b)
(Fig.
2)
[3].
The
structures
of
the
isolated
compounds
were
determined
on
the
basis
of
extensive
analyses
of
their
spectroscopic
data
(NMR,
MS,
and
UV).
All
the
derivatives
exhibited
potent
antibacte-‐
rial
activity
against
E.
coli,
and
antifungal
activity
against
C.
albicans
and
A.
brasiliensis
with
MIC
<
5
ppm.
Figure
1.
Caulerpenyne
from
Caulerpa
taxifolia.
Figure
2.
New
p-‐sulfooxyphenylpyruvic
acid
methyl
ester
(1a
and
1b)
from
Caulerpa
taxifolia.
Acknowledgements:
E.
Sfecci
is
the
recipient
of
a
thesis
grant
from
the
"Conseil
Régional
Provence
Alpes
Côte
d’azur"
and
a
travel
grant
from
the
“Association
Lapervenchelif”.
M.
Mehiri
research
is
supported
by
the
french
program
ENVI-‐Med
"MEDIBIO",
the
ANR/Investissements
d’Avenir
program
via
the
OCEANOMICs
project
(grant
#ANR-‐11-‐BTBR-‐0008),
and
the
H2020
European
program
via
the
EMBRIC
project.
Keyword: Caulerpa taxifolia.
References:
[1] Amico
V,
Oriente
G,
Piattelli
M,
Tringali
C,
Fattorusso
E,
Magno
S,
Mayol
L.
Caulerpenyne,
an
unusual
sequiterpenoid
from
the
green
alga
Caulerpa
prolifera.
Tetrahedron
Lett
1978;
19:
3593–3596
[2] Hodgson
LM.
Antimicrobial
and
antineoplastic
activity
in
some
south
Florida
seaweeds.
Bot
Mar
1984;
27:
387–390
[3] Mancini
I,
Guella,
G,
Defant,
A,
Candenas,
ML,
Armesto,
CP,
Depentori,
D,
Pietra,
F.
Polar
metabolites
of
the
tropical
green
seaweed
Caulerpa
taxifolia
which
is
spreading
in
the
Mediterranean
sea:
glycoglycerolipids
and
stable
enols
(α-‐keto
esters).
Helv
Chim
Acta,
1998;
81:
1681–1691
P565
Bioassay-‐guided
identification
of
bioactive
protein
compounds
from
the
Mediterranean
demosponge
Tethya
meloni
Eugenia
Gentile1,
Carlotta
Nonnis
Marzano2,
Tiziana
Latronico1,
Anna
Fasano1,
Rocco
Ros-‐
sano3,
Giuseppe
Corriero2,
Grazia
Maria
Liuzzi1
1Department
of
Biosciences,
Biotechnologies
and
Biopharmaceutics,
University
of
Bari
“Aldo
Moro”,
Via
Orabona
4,
70125
Bari,
Italy;
2Department
of
Biology,
University
of
Bari
“Aldo
Moro”,
Via
Orabona
4,
70125
Bari,
Italy;
3Department
of
Sciences,
University
of
Basilicata,
Via
dell'Ateneo
Lucano
10,
85100
Potenza,
Italy
Marine
natural
products
extracted
from
sponges
represent
a
new
source
for
drug
discovery.
Among
marine
organisms,
the
phylum
Porifera
is
the
most
prolific
for
the
production
of
pharmacologically
active
compounds,
which
are
characterized
by
unique
chemical
structures
[1].
Potential
drug
discovery
targets
include
matrix
metalloproteinases
(MMPs),
and
in
par-‐
ticular
gelatinases
A
(MMP-‐2)
and
B
(MMP-‐9),
which
are
involved
at
different
levels
in
the
pathogenesis
of
several
human
diseases
[2].
We
used
a
simple
method
for
preparing
aqueous
extracts
from
the
Mediterranean
demosponge
Tethya
meloni
[3],
which
allowed
the
extrac-‐
tion
of
water-‐soluble
compounds
by
homogenization
of
sponge
tissue
in
phosphate
buffered
saline
[4].
The
analysis
by
SDS-‐PAGE
showed
a
very
rich
protein
pattern.
Using
a
well-‐known
in
vitro
model
represented
by
lipopolysaccharides
(LPS)-‐activated
rat
astrocytes,
we
demon-‐
strated
the
capability
of
the
sponge
extract
to
inhibit
the
activity
of
MMP-‐2
and
MMP-‐9
in
a
dose-‐dependent
manner.
In
particular,
astrocytes
treated
with
the
crude
extract
at
the
higher
non-‐cytotoxic
concentration
of
60
μg/ml
showed
a
50%
inhibition
of
MMP-‐2
and
a
70%
inhi-‐
bition
of
MMP-‐9
in
comparison
with
the
positive
control
(LPS-‐activated
astrocytes).
To
identi-‐
fy
the
bioactive
compounds
with
anti-‐MMP
activity,
the
crude
extract
was
subjected
to
anion-‐
exchange
chromatography
and
the
pools
obtained
by
the
different
protein
peaks
were
tested
on
LPS-‐activated
astrocytes.
We
identified
one
fraction,
which
contains
a
major
protein
band
of
30
kDa,
which
exhibited
an
80%
inhibition
towards
both
MMP-‐2
and
MMP-‐9,
in
comparison
to
positive
control.
In
an
effort
aimed
at
the
identification
of
the
bioactive
compounds
with
anti-‐MMP
activity,
this
fraction
was
subjected
to
2-‐D
electrophoresis.
Results
indicated
the
presence
of
about
10
spots
at
pI
between
4.5
and
9.5;
probably
isoforms
of
the
same
protein.
The
identification
of
protein,
which
might
be
used
for
pharmaceutical
applications,
is
now
in
course
by
MS-‐MS
analysis.
Acknowledgements:
Dr.
Frine
Cardone
is
acknowledged
for
sample
collection,
Dr.
Marilena
Larocca
is
acknowledged
for
technical
support
in
the
2-‐DE
analyses.
Keywords: Demosponge, aqueous extracts, bioactive compounds, matrix metalloproteinases
References:
[1] Mehbub
MF,
Lei
J,
Franco
C,
Zhang
W.
Marine
sponge
derived
natural
products
between
2001
and
2010:
trends
and
opportunities
for
discovery
of
bioactives.
Mar
Drugs
2014;
12:
4539−4577
[2] Pulkoski-‐Gross
AE.
Historical
perspective
of
matrix
metalloproteases.
Front
Biosci
2015;
7:
125−149
[3] Corriero
G,
Gadaleta
F,
Bavestrello
G.
A
new
Mediterranean
species
of
Tethya
(Porifera:
Tethyida:
Demospongiae).
Ital
J
Zool
2015;
82:
1−9
[4] Di
Bari
G,
Gentile
E,
Latronico
T,
Corriero
G,
Fasano
A,
Nonnis
Marzano
C,
Liuzzi
GM.
Com-‐
parative
analysis
of
protein
profiles
of
aqueous
extracts
from
marine
sponges
an-‐
dassessment
of
cytotoxicity
on
different
mammalian
cell
types.
Environ
Tox
Pharm
2014;
38:
1007−1015
P566
Oxymatrine
and
matrine
biotransformation
by
microorganisms
and
the
evaluation
of
effects
on
5-‐lipoxygenase
inhibition
Fatih
Demirci1,2,
Ayşe
Esra
Karadağ3,4,
Mustafa
Güzel5
1
Anadolu
University,
Faculty
of
Pharmacy,
Department
of
Pharmacognosy,
2
Faculty
of
Health
Sciences,
26470-‐Eskişehir,
3
Graduate
School
of
Health
Sciences;
4
Medipol
University,
Faculty
of
Pharmacy,
De-‐
partment
of
Pharmacognosy;
5
Faculty
of
Internal
Medicine,
Department
of
Medical
Pharmacology,
34810,
Kavacık/Beykoz,
İstanbul,
Turkey
Sophora
L.
species
of
Fabaceae
contain
bioactive
quinolizidine
type
alkaloids
such
as
ox-‐
ymatrine
and
matrine.
It
is
well
documented
that
some
Sophora
species
are
used
in
Tradition-‐
al
Chinese
Medicines.
The
mechanisms
of
inflammation
involve
complex
cascades
of
events
where
chemical
messengers
play
an
important
role.
An
important
class
of
messengers
are
the
pro-‐inflammatory
leukotrienes
resulting
from
the
metabolism
of
arachidonic
acid,
which
be-‐
gins
with
its
oxidation
by
the
enzyme
5-‐lipoxygenase
[1].
The
aim
of
this
study
was
to
synthe-‐
size
derivatives
of
oxymatrine
and
matrine
by
microbial
biotransformation.
As
a
result,
ox-‐
ymatrine
was
metabolized
to
its
deoxy-‐analogue
matrine,
by
Fusarium
solani,
Saccharomyces
cerevisiae,
Mucor
ramannianus,
Alternaria
alternata
and
Pycnoporus
cinnabarinus
among
15
different
microorganisms
evaluated.
The
metabolites
were
evaluated
by
LC-‐MS/MS.
In
the
time-‐based
evaluation
of
the
transformation,
complete
metabolization
of
oxymatrine
was
ob-‐
served
after
48
hours
of
incubation.
The
substrate
matrine
was
metabolized
into
two
different
analogues
by
M.
ramannianus.
Main
metabolite
was
tentatively
assigned
as
hydroxy-‐
derivative
of
matrine,
where
the
other
was
identified
as
an
aromatized
derivative.
Additional-‐
ly,
both
the
substrate
and
metabolites
were
evaluated
in
vitro
for
their
lipoxygenase
(5-‐LOX)
inhibitory
activity
by
means
of
spectrophotometric
methods.
As
a
result,
oxymatrine
showed
an
inhibition
of
21.1%,
where
matrine
exhibited
10.5%,
respectively.
The
metabolites
showed
no
inhibition
at
the
tested
concentrations.
To
the
best
of
our
knowledge
this
is
the
first
bio-‐
transformation
of
oxymatrine
and
matrine
by
the
above-‐mentioned
microorganisms
and
LOX
evaluation.
The
biological
evaluation
of
matrine
derivatives
is
ongoing.
Acknowledgement:
This
work,
which
is
part
of
the
MSc
thesis
of
AEK
was
supported
by
Anadolu
Universi-‐
ty,
Scientific
Research
Projects
Commission
Fund
Projects
grant
no:
1601S029
Keywords: Matrine, oxymatrine, microbial biotransformation, LC-‐MS/MS, lipoxygenase in-‐
hibitor activity
References:
[1] Baylac
S,
Racine
P.
Inhibition
of
5-‐lipoxygenase
by
essential
oils
and
other
natural
fragrant
extracts.
Int
J
Aromather
2003;
13:
138-‐142
P567
The
effects
of
brown
macro
algae
from
Northen
coasts
of
Persian
Gulf
on
melanogenesis
Foroogh
Namjoyan1,
Mojtaba
Alishahi2,
Alireza
Jahangiri3,
Massoumeh
Farasat1,
Hamideh
Mousavi4
1
Marine
Pharmaceutical
Research
Center,
Pharmacognosy
Department,
School
of
Pharmacy,
Ahvaz
Jun-‐
dishapur
University
of
Medical
Sciences,
Iran,
2
Faculty
of
veterinary
Medicine,
Shahid
Chamran
Universi-‐
ty,
Ahvaz,
Iran,
3
Medicinal
Chemistry
Department,
Ahvaz
Jundishapur
University
of
Medical
Sciences,
Ahvaz,
Iran,
4
Faculty
of
Pharmacy,
Ahvaz
Jundishapur
University
of
Medical
Sciences,
Iran
Melanin
is
the
most
important
factor
determining
skin
color
and
is
biosynthesized
by
enzyma-‐
tic
and
non-‐enzymatic
reactions.
Tyrosinase
is
a
key
and
rate-‐limiting
enzyme
in
melanin
production
which
catalyzes
tyrosine
to
L-‐Dopa
and
them
to
dopaquinone.
Therefore
tyrosi-‐
nase
inhibitors
could
be
utilized
in
treatment
of
abnormal
pigmentation
disorders
and
as
skin-‐whitening
agents
in
the
cosmetic
industry.
Due
to
adverse
effects
of
synthetic
drugs
and
public
desire
to
natural
product,
in
this
study
we
selected
brown
macro
algae
from
Persian
Gulf
to
evaluate
their
anti
tyrosinase
and
anti
melanogenesis
activities.
17
brown
macro
algae
from
Persian
Gulf
were
collected.
Their
methanolic
extracts
were
used
for
spectrophotometric
analysis
of
inhibitory
effects
on
diphenolase
activity
of
mushroom
tyrosinase
in
concentra-‐
tions
100,
250
and
500
µg/mL
using L-‐Dopa
as
substrate.
Then
we
evaluated
activities
of
ma-‐
cro
algae
with
high
inhibition
potency
on
hydroxylation
of
L-‐tyrosine
by
mushroom
and
zebra
fish
tyrosinase.
Anti-‐melanogenesis
effects
of
algae
were
studied
on
zebra
fish
as
an
alternati-‐
ve
in
vivo
model
at
100
µg/mL.
Kojic
acid
was
used
as
positive
control.
Padina
boergesenii,
Colpomenia
sinuosa
and
Sargassum
swartzii
showed
the
highest
inhibitory
activities
among
tested
samples
on
mushroom
tyrosinase
(51.75,
23.25
and
20.55
%
at
500
µg/mL,
respecti-‐
vely).
They
inhibited
zebra
fish
tyrosinase
more
potent
than
kojic
acid
(83,
43
and
24%,
res-‐
pectively
vs
50%
inhibition
for
kojic
acid).
Moreover,
the
algae
reduced
melanin
synthesis
in
zebra
fish
42,
28
and
20%,
respectively
vs
50%
for
kojic
acid.
These
results
and
other
repor-‐
ted
skin
care
effects
of
Padina
boergesenii,
Colpomenia
sinuosa
and
Sargassum
swartzii
provi-‐
ded
that
they
have
high
potential
for
using
in
the
cosmetics
and
medicinal
formulations
deve-‐
lopment
to
prevent
and
treat
hyperpigmentation
diseases.
Thus,
more
studies
on
other
me-‐
chanisms
of
melanogenesis
inhibition
and
related
active
compounds
of
the
algae
are
required.
Key
words:
Persian
Gulf,
algae,
melanogenesis,
mushroom
tyrosinase,
zebra
fish,
Padina
boergesenii
P568
The
effects
of
red
macro
algae
from
Northen
coasts
of
Persian
Gulf
on
melanogenesis
Foroogh
Namjoyan1,
Mojtaba
Alishahi2,
Alireza
Jahangiri3,
Massoumeh
Farasat1,
Hamideh
Mousavi4
1
Marine
Pharmaceutical
Research
Center,
Pharmacognosy
Department,
School
of
Pharmacy,
Ahvaz
Jun-‐
dishapur
University
of
Medical
Sciences,
Iran,
2
Faculty
of
veterinary
Medicine,
Shahid
Chamran
Universi-‐
ty,
Ahvaz,
Iran,
3
Medicinal
Chemistry
Department,
Ahvaz
Jundishapur
University
of
Medical
Sciences,
Ahvaz,
Iran,
4
Faculty
of
Pharmacy,
Ahvaz
Jundishapur
University
of
Medical
Sciences,
Iran
Tyrosinase
is
a
key
enzyme
in
melanin
production
and
catalyzes
hydroxylation
of
L-‐tyrosine
to
L-‐Dopa
then,
oxidation
of
L-‐Dopa
to
dopaquinone.
Therefore,
tyrosinase
inhibitors
are
used
in
cosmetic
and
medicinal
industries
to
prevent
or
treat
overproduction
of
melanin
diseases
in-‐
cluding,
melasma,
solar
lentigo
and
post
inflammatory
melanoderma.
According
to
public
de-‐
sire
to
natural
whitening
agents,
in
this
study
we
selected
red
macro
algae
from
Persian
Gulf
to
evaluate
their
anti-‐tyrosinase
and
anti-‐melanogenesis
activities.
3
red
macro
algae
from
Per-‐
sian
Gulf
including,
Digenea
simplex,
Laurencia
pappilosa
and
Laurencia
paniculata
were
col-‐
lected.
The
effects
of
methanolic
algal
extracts
on
diphenolase
activity
of
mushroom
tyrosinase
were
studied
in
concentrations
100,
250
and
500
µg/mL
using
L-‐Dopa
as
substrate.
Then,
we
evaluated
activity
of
macroalgae
with
high
inhibition
potency
on
hydroxylation
of
L-‐tyrosine
by
mushroom
and
zebra
fish
tyrosinase.
Anti-‐melanogenesis
effects
of
algae
were
studied
on
zebra
fish
as
an
alternative
in
vivo
model
at
100
µg/mL.
Kojic
acid
was
used
as
positive
control.
The
red
macro
algae
inhibited
significantly
diphenolase
activity
of
mushroom
tyrosinase
weaker
than
kojic
acid
(32,
18
and
11%,
respectively
for
D.
simplex,
L.
pappilosa
and
L.
panicu-‐
lata
at
500
µg/mL).
D.
simplex
showed
the
most
anti-‐tyriosinase
activity
among
algal
samples.
It
exhibited
significantly
weaker
anti-‐monophenolase
tyrosinase
activity
than
kojic
acid.
D.
simplex
inhibited
tyrosinase
activity
and
total
melanin
of
zebra
fish
model
43.18%
and
47.27%,
respectively
and
Kojic
acid
reduced
50.45%
and
50.21%,
respectively.
These
results
and
other
skin
care
effects
of
D.
simplex
provided
that
it
could
be
used
as
ingredients
for
whit-‐
ing
cosmetics.
More
studies
on
its
active
compounds
could
lead
to
discovery
of
new
whitening
compounds
for
development
of
skin
care
products.
Keywords:
Algae,
Persian
Gulf,
melanogenesis,
mushroom
tyrosinase,
zebra
fish
P569
Potential
antimicrobial
activities
of
seagrasses,
Zostera
spp.
from
Aegean
Sea
against
Vibrio
alginolyticus
ATCC
17749
Hijran
Yavuzcan1,
Müjde
Eryılmaz2,
Özlem
Bahadır
Acıkara3,
Serkan
Özbilgin3,
Burçin
Ergene1,
Sertel
Seçer3,
Gülçin
Saltan
İşcan3
1
Department
of
Fisheries
and
Aquaculture,
Ankara
University,
Faculty
of
Agriculture,
06110
and
Ankara,
Turkey,
2
Department
of
Pharmaceutical
Microbiology,
Ankara
University,
Faculty
of
Pharmacy,
06100
and
Ankara,
Turkey,
3
Department
of
Pharmacognosy,
Ankara
University,
Faculty
of
Pharmacy,
06100
and
Ankara,
Turkey
The
study
of
marine
organisms
for
their
bioactive
potential
has
gained
momentum
in
recent
years
with
a
growing
recognition
of
their
importance
to
human
health.
Bioactive
chemicals
from
marine
plants,
such
as
phenolic
compounds
that
play
important
roles
for
their
survival
and
growth,
are
now
explored
for
the
development
of
new
drugs
and
health
products
for
hu-‐
mans.
Zostera
spp.
is
one
of
the
most
widely
distributed
plant
genera
in
marine
environment,
and
is
comprises
important
species
in
coastal
ecosystems
by
contributing
to
nutrient
cycling
and
sediment
stabilization,
and
providing
food
stuffs
and
habitat
for
many
marine
organisms.
The
aim
of
this
study
is
to
evaluate
the
antimicrobial
activity
of
extracts
of
Zostera
spp.
collec-‐
ted
from
Aegean
Sea
Coasts
against
Vibrio
alginolyticus
ATCC
17749.
Dead
leaves
of
Zostera
marina
and
Z.
noltii
were
collected
from
Urla
district
near
Izmir
Bay
(Lat
380
31’
N
and
Long
260
69’
E)
in
autumn.
Taxononomic
identity
of
the
plants
was
con-‐
firmed
by
reference
to
literature.
Extracts
were
prepared
by
maceration
at
room
temperature
succesively
using
n-‐hexane,
ethylacetate
and
methanol.
All
solvents
were
evaporated
under
vacuum
and
the
dried
residues
were
dissolved
in
DMSO/water
(20%
v/v)
before
testing
for
their
antimicrobial
activities.
The
fractions
were
screened
in
vitro
for
their
potential
antibac-‐
terial
activities
against
Vibrio
alginolyticus
ATCC
17749.
Micro
broth
dilution
method
was
used
for
determination
of
the
minimum
inhibitory
concentrations
(MIC).
The
cultures
were
obtained
in
Mueller
Hinton
Broth
(Difco,
Difco
Laboratories,
Detroit,
MI,
USA)
supplemented
with
2%
NaCl.
Serial
two-‐fold
dilutions
ranging
from
10
to
0.078
mg/ml
were
prepared
in
medium.
A
set
of
wells
containing
only
inoculated
broth
were
used
as
control.
After
incuba-‐
tion
for
18-‐24
h
at
35±1°C,
the
last
well
with
no
microbial
growth
was
recorded
to
represent
MIC
value
(mg/ml).
The
MIC
results
of
the
tested
extracts
are
shown
in
Table
1.
Ethyl
acetate
extract
of
Zostera
spp.
exhibited
better
antimicrobial
activity
than
n-‐hexane
and
methanol
extracts.
Table
1.
MIC
values
(mg/mL)
of
Zostera
spp.
against
Vibrio
alginolyticus
ATCC
17749
Test
Microorganism
Extracts
n-‐hexane
Ethylacetate
Methanol
Vibrio
alginolyticus
ATCC
17749
10
0.625
10
Acknowledgements:
This
study
was
supported
by
TUBITAK
(The
Scientific
and
Technical
Research
Coun-‐
cil
of
Turkey)
(Project
Number:
114Y141).
Keywords: Seagrasses, Zostera spp., antimicrobial activity
P570
Keikipukalides:
Furanocembranoid
aldehyde
derivatives
pro-‐
duced
by
the
Antarctic
deep
sea
soft
coral
Plumarella
delicatissi-‐
ma
Jacqueline
L.
von
Salm1,2,
Shane
Clark1,
Prasanth
Nemani1,
Steve
Ferlita1,
Nerida
G.
Wilson3,
Bill
J.
Baker1,2
1
Department
of
Chemistry
and
2
Center
for
Drug
Discovery
and
Innovation,
University
of
South
Florida,
4202
E.
Fowler
Ave.,
Tampa,
FL
33620,
USA,
3
Western
Australia
Museum,
49
Kew
St.
Welshpool,
Perth,
WA
6106,
Australia,
†current
address:
Department
of
Chemistry,
Simon
Fraser
University,
8888
University
Dr.,
Burnaby,
BC
V5A
1S6,
Canada
Here
we
present
the
isolation
and
characterization
of
five
new
furanocembranoid
diterpenes,
the
keikipukalides
A-‐E,
as
well
as
the
known
pukalide
derivative,
pukalide
aldehyde.
These
compounds
were
extracted
from
the
deep
cold-‐water
octocoral
species,
Plumarella
delicatis-‐
sima,
in
the
Southern
Ocean
along
the
Scotia
Arc.
They
most
resemble
the
neuromuscular
tox-‐
ins
lophotoxin
[1],
pukalide
[2],
and
leptolide
[3].
Structure
elucidation
was
accomplished
using
various
spectroscopic
techniques
including
one-‐
and
two-‐dimensional
NMR
spectrosco-‐
py,
mass
spectrometry,
and
X-‐ray
crystallography.
Biological
activity
of
these
scaffolds
is
well
known
to
have
negative
effects
on
the
neurological
system
of
vertebrates
by
irreversibly
bind-‐
ing
to
acetyl
choline
nicotinic
receptors.
The
crude
extract
had
minor
cytotoxicity
towards
the
development
of
sea
urchin
embryos.
However,
screening
against
the
ESKAPE
bacterial
cell
lines
identified
no
activity.
Keywords:
Antarctica,
marine
natural
products,
deep-‐sea
corals,
diterpenes,
NMR
Spectros-‐
copy
References:
[1] Fenical
W,
Okuda
RK,
Bandurraga
MM,
Culver
P,
Jacobs
RS.
Lophotoxin:
a
novel
neuromus-‐
cular
toxin
from
Pacific
sea
whips
of
the
genus
Lophogorgia.
Science
1981;
212:
1512-‐
1514
[2] Missakian
MG,
Burreson
BJ,
Scheuer
PJ.
Pukalide,
a
furanocembranolide
from
the
soft
coral
Sinularia
abrupta.
Tetrahedron
1975;
31:
2513-‐2515
[3] Gutiérrez
M,
Capson
TL,
Guzmán
HM,
González
J,
Ortega-‐Barría
E,
Quinoá
E,
Riguera
R.
Lep-‐
tolide,
a
new
furanocembranolide
diterpene
from
Leptogorgia
alba.
J
Nat
Prod
2005;
68:
614-‐616
P571
Protective
effect
of
Oyster
hydrolysate
peptide
in
alcohol
induced
alcoholic
fatty
liver
in
SD-‐rats
Jae-‐Hyuk
Byun1,
Yeung-‐Joon
Choi3,
Se-‐Young
Choung1,2
1Department
of
Life
and
Nanopharmaceutical
Sciences,
College
of
Pharmacy,
Kyung
Hee
University,
Hoegi-‐dong,
Dongdaemun-‐gu,
Seoul
130-‐701,
Republic
of
Korea,
2
Department
of
Preventive
Pharmacy
and
Toxicology,
College
of
Pharmacy,
Kyung
Hee
University,
Hoegi-‐dong,
Dongdaemun-‐gu,
Seoul
130-‐701,
Republic
of
Korea,
3Department
of
Seafood
Science
and
Technology/Institute
of
Marine
Industry,
Gyeong-‐
sang
National
University,
Gyeongnam
650-‐160,
Republic
of
Korea
In
the
present
study,
protective
effects
of
Oyster
hydrolysate
peptide
(TGPN)
were
investigat-‐
ed
against
alcoholic
liver
in
Sprague-‐Dawley
rats.
Five
week
old
Sprague-‐Dawley
male
rats
were
divided
into
two
groups
and
fed
Lieber-‐DeCarli
diet
containing
5%
(w/v)
alcohol
(ED)
or
an
isocaloric
amount
of
dextrin-‐maltose
in
the
controls
(ND)
for
4
weeks
[1].
Then
ED
were
treated
with
TGPN
(50,
100,
or
200
mg/kg)
or
silymarin
(100mg/kg)
for
another
4
weeks.
The
TGPN
supplementation
significantly
lowered
serum
lipid
levels
(TG,
TC,
FFA),
he-‐
patic
lipid
levels
(TG,
TC,
FFA),
liver-‐index
and
hepatotoxicity
(ALT,
AST)
in
ED-‐induced
rats.
Histological
analysis
indicated
that
the
TGPN-‐treated
group
showed
lowered
number
of
lipid
droplets,
size
of
adipocytes,
and
reduced
hepatic
lipid
amount
compared
to
the
ED
group.
To
understand
the
molecular
mechanism
of
TGPN,
the
expression
of
genes
involved
in
alcoholic
fatty
liver
was
measured.
The
expression
of
fatty
acid
oxidation
and
thermogenesis-‐related
genes
(PPAR-‐α,
CPT-‐1)
of
ED-‐induced
rats
[2]
was
increased
by
TGPN.
On
the
other
hand,
TGPN
treatment
resulted
in
decreased
expression
levels
of
lipogenesis
related
genes
such
as
SREBP-‐1c,
FAS,
and
SCD1.
These
results
suggest
that
TGPN
may
have
ability
to
ameliorate
al-‐
coholic
fatty
liver
and
hepatic
protection.
Acknowledgements:
This
study
was
supported
by
the
Korea
Institute
of
Marine
Science
&
Technology
promotion.
Keywords: Oyster hydrolysate, Alcoholic fatty liver, Lipid metabolism, SD-‐rat
References:
[1] Lieber,
CS,
Decarli,
LM,
Animal
models
of
chronic
ethanol
toxicity.
Methods
Enzym
1994;
233:
585–594.
[2] Chang
CJ,
Tzeng
TF,
Liou
SS,
Chang
YS,
Liu
IM.
Kaempferol
regulates
the
lipid-‐profile
in
high-‐fat
diet-‐fed
rats
through
an
increase
in
hepatic
PPARα
levels.
Planta
Med
2011;
77:
1876-‐1882
P572
FITC-‐labeled
dieckol
acting
as
endoplasmic
reticulum
stress
sig-‐
naling
inhibitor
in
RAW
264.7
macrophage
Jong
Hwan
Kwak1,
Jwa-‐Jin
Kim2,
Kyung
Bok
Lee2,
Yung
Choon
Yoo2,
Jong
Seung
Kim3
1
School
of
Pharmacy,
Sungkyunkwan
University,
Suwon
440-‐746,
Korea,
2
College
of
Medicine,
Konyang
University,
Daejeon
302-‐718,
Korea,
3
Department
of
Chemistry,
Korea
University,
Seoul
136-‐701,
Korea
The
endoplasmic
reticulum
(ER)
is
an
important
organelle
for
protein
synthesis,
protein
fold-‐
ing,
lipid
biosynthesis,
calcium
storage
and
homeostasis.
It
plays
a
central
role
in
the
co-‐
and
post-‐translational
protein
processing,
including
disulfide
bond
formation
and
N-‐linked
glyco-‐
sylation,
which
are
essential
for
proper
protein
folding.
ER
stress
and/or
sustained
activation
of
the
unfolded
protein
response
(UPR)
have
been
implicated
in
some
chronic
inflammatory
diseases,
and
the
development
of
neurodegenerative
disorders
such
as
Alzheimer’s
disease
and
Parkinson’s
disease
[1].
Dieckol,
a
phlorotannin,
has
been
isolated
from
several
brown
algae
of
genus
Eisenia
and
Ecklonia
and
reported
to
show
various
biological
activities
[2].
We
introduced
fluorone
onto
dieckol
to
investigate
its
ER
stress
suppressing
activity
and
localiza-‐
tion
within
living
cells.
The
fluorescein
isothiocyanate
(FITC)-‐labeled
dieckol
(1)
was
synthe-‐
sized
through
a
click
reaction.
Early
changes
in
levels
of
well-‐known
ER
stress
markers,
in-‐
cluding
the
C/EBP
homologous
protein
(CHOP)
and
transcription
factor
X-‐box
binding
protein
1
(XBP1),
were
evaluated.
Compound
1
significantly
reversed
the
LPS
treatment-‐induced
in-‐
crease
in
CHOP
and
XBP1
levels
in
RAW
264.7
cells.
To
investigate
the
intracellular
localiza-‐
tion
of
1,
colocalization
experiments
were
implemented
using
confocal
laser
microscopy,
indi-‐
cating
that
compound
1
is
located
mainly
in
the
ER
of
RAW
264.7
cells.
Acknowledgements:
This
work
was
supported
by
a
grant
from
Marine
Biotechnology
Program
(PJT200620,
Genome
Analysis
of
Marine
Organisms
and
Development
of
Functional
Applications)
funded
by
Ministry
of
Oceans
and
Fisheries.
Keywords: FITC-‐labeled dieckol, ER stress, CHOP, XBP1, intracellular localization
References:
[1] Kwak
JH,
Yang
Z,
Yoon
B,
He
Y,
Uhm
S,
Shin
H-‐C,
Lee
BH,
Yoo
YC,
Lee
KB,
Han
S-‐Y,
Kim
JS.
Blood-‐brain
barrier-‐permeable
fluorone-‐labeled
dieckols
acting
as
neuronal
ER
stress
signalling
inhibitors.
Biomaterials
2015;
61:
52-‐60
[2] Kwak
JH,
He
Y,
Yoon
B,
Koo
S,
Yang
Z,
Kang
EJ,
Lee
BH,
Han
S-‐Y,
Yoo
YC,
Lee
KB,
Kim
JS.
Syn-‐
thesis
of
rhodamine-‐labelled
dieckol:
its
unique
intracellular
localization
and
potent
anti-‐
inflammatory
activity.
Chem
Commun
2014;
50:
13045-‐13048
P573
Steroids
and
cembranoids
from
the
soft
corals
Nephthea
erecta
and
Lobophytum
crissum
Jui-‐Hsin
Su1,
Bo-‐Rong
Peng1,
Shih-‐Kai
Chou2,
Ming-‐Cheng
Shih2,
Yu-‐Ting
Huang3
1
National
Museum
of
Marine
Biology
and
Aquarium
and
Graduate
Institute
of
Marine
Biology,
National
Dong
Hwa
University,
Pingtung
944,
Taiwan,
2
Department
of
Biological
Science
and
Technology,
I-‐Shou
University,
Kaohsiung
824,
Taiwan,
3
Department
of
Nutrition,
I-‐Shou
University,
Kaohsiung
824,
Taiwan
One
new
10-‐demethylated
steroid,
nephtheasteroid
A
(1),
one
new
19-‐oxygenated
steroid,
nephtheasteroid
B
(2),
and
four
known
steroids
(3–6)
were
isolated
from
the
wild-‐type
soft
coral
Nephthea
erecta.
Two
new
cembranoids,
culobophylins
D
(7)
and
E
(8),
along
with
eight
known
compounds
(9
–16)
were
isolated
from
the
cultured
soft
coral
Lobophytum
crassum.
The
structures
were
elucidated
by
means
of
IR,
MS,
and
NMR
techniques,
and
comparison
of
the
NMR
data
with
those
of
known
analogues.
Compound
1
was
found
to
have
a
novel
skele-‐
ton
with
the
C10
demethylation
of
the
normal
steroid.
Evaluation
of
the
cytotoxicities
showed
that
compound
9,
the
most
potent
of
compounds
1–16,
exhibited
cytotoxicity
against
the
K562,
Molt-‐4,
U937,
Sup-‐T1
and
Ca9-‐22
cancer
cell
lines
with
IC50
values
of
1.2,
0.4,
2.4,
0.5
and
1.1
μg/mL,
respectively.
Moreover,
compound
10
also
showed
significant
U937
and
Sup-‐
T1
inhibitory
activity,
with
IC50
values
of
1.87
and
1.13
μg/mL.
Acknowledgements: the Ministry of Science and Technology, Taiwan
Keywords: Steroids, soft coral, Nephthea erecta, Lobophytum crassum, cytotoxicity
P574
Bioactive
chemical
constituents
from
the
Formosan
red
algae
Laurencia
tristicha
Jia-‐Yu
Chen1,
Chiung-‐Yao
Huang
1,
Jyh-‐Horng
Sheu
1,2
1
Department
of
Marine
Biotechnology
and
Resources,
National
Sun
Yat-‐sen
University,
804,
Kaohsiung
Taiwan,
2
Frontier
Center
for
Ocean
Science
and
Technology,
National
Sun
Yat-‐sen
University,
804,
Kaohsiung
Taiwan
Secondary
metabolites
discovered
from
red
alga
are
predominantly
sesquiterpenoids
and
usually
chlorinated
or
brominated
[1].
In
our
continuing
investigation
on
the
chemical
con-‐
stituents
of
marine
alga
of
the
genus
Laurencia
[2]
red
algae
L.
tristicha
was
collected
and
studied
in
order
to
discover
bioactive
marine
natural
products
from
Formosa
red
alga.
This
investigation
led
to
the
discovery
of
eight
new
halogenated
chamigrane-‐type
sesquiterpenoids
1–8
and
one
new
bromocuparane-‐type
sesquiterpene
9,
along
with
nine
known
compounds
10–18.
The
structure
of
a
known
compound,
ma’iliohydrin
(15)
[3],
was
revised
too.
Both
compounds
10
and
11
were
discovered
from
nature
for
the
first
time.
The
absolute
configura-‐
tion
of
12
was
confirmed
by
single-‐crystal
X-‐ray
diffraction
analysis.
The
anti-‐inflammatory
activities
of
compounds
1–18
on
neutrophil
pro-‐inflammatory
responses
were
evaluated
by
measuring
their
ability
in
suppressing
fMLP/CB-‐induced
superoxide
anion
(O2−
•)
generation
and
elastase
release
in
human
neutrophils.
From
the
results,
allo-‐isoobtusol
(17)
[4]
showed
strong
inhibitions
(44.44
± 4.02
%)
toward
elastase
release
generation
at
10
µM.
OH OH
14 O O
13 1 O O 15
1
3
Br B Br Br R
7 4 Br
6 4 6
A
9 11
HO HO HO HO
12 O
1 2 3 R = CHBr2 5
4 R = CH2OH
Cl Br 15
Br 8
OMe
12 13
Br OH 6
1 10
Br
3 14
HO 5
O O
HO
6 7 8 9
Acknowledgements:
Tsong-‐Long
Hwang
and
Shu-‐Fen
Chiou
are
acknowledged
for
anti-‐inflammatory
and
antibacterial
technical
assistance
Keywords: Red algae, Laurencia tristicha, sesquiterpenoid
References:
[1] Wang
BG,
Gloer
JB,
Ji
NY,
ZhaoJC.
Halogenated
Organic
Molecules
of
Rhodomelaceae
Origin:
Chemistry
and
Biology.
Chem
Rev
2013;
113:
3632–3685
[2] Fang
HY,
Chiou
SF,
Uvarani
C,
Wen
ZH,
Hsu
CH,
Wu
YC,
Wang
WL,
Liaw
CC,
Sheu
JH.
Cyto-‐
toxic,
anti-‐inflammatory,
and
antibacterial
sulfur-‐containing
polybromoindoles
from
the
Formosan
red
alga
Laurencia
brongniartii.
Bull
Chem
Soc
Jpn
2014;
87:
1278–1280
[3] Francisco
MEY,
Erickson
KL.
Ma'iliohydrin,
a
cytotoxic
chamigrene
dibromohydrin
from
a
Philippine
Laurencia
species.
J
Nat
Prod
2001;
64:
790–791
[4] Francisco
MEY,
Turnbull
MM,
Erickson
KL.
Cartilagineol,
the
fourth
lineage
of
Laurencia-‐
derived
polyhalogenated
chamigrene.
Tetrahedron
Lett
1998;
39:
5289–5292
P575
Structure
determination
and
biosynthesis
of
the
antifungal
bu-‐
tyrolactols
from
Streptomyces
sp.
Keebeom
Ko1,
Hui-‐Ming
Ge2,
Jongheon
Shin1,
Dong-‐Chan
Oh1
1Natural
Products
Research
Institute,
College
of
Pharmacy,
Seoul
National
University,
1
Gwanak-‐ro,
Gwa-‐
nak-‐gu,
Seoul,
08826,
Republic
of
Korea
2School
of
Life
Science,
Nanjing
University,
Nanjing
210046,
Chi-‐
na
Marine
actinomycetes
have
been
drawing
attention
as
prolific
sources
of
new
bioactive
sec-‐
ondary
metabolites
[1].
In
our
research
for
new
bioactive
secondary
metabolites
form
marine
microorganisms,
we
selectively
isolated
actinomycete
strains
from
marine
sediment
samples
from
the
western
area
of
Jeju
Island
in
Republic
of
Korea
and
screened
their
secondary
me-‐
tabolites
profiles
by
LC/MS.
During
the
chemical
profile
analysis,
we
observed
that
one
of
the
actinomycete
strains
named
TH11
(Streptomyces
sp.)
produces
previously
reported
antifungal
butyrolactols
A
and
B,
which
bear
a
seven
consecutive
1,2-‐diol
moiety
[2],
and
their
new
de-‐
rivatives,
butyrolactols
C
and
D.
Because
the
absolute
configuration
of
butyrolactol
A
has
not
previously
determined,
we
established
the
stereochemistry
of
butyrolactol
A
by
J-‐based
con-‐
figuration
analysis
using
ROESY
and
HETLOC
data
and
Mosher’s
method.
Biosynthetically
in-‐
terestingly,
four
oxygen-‐bearing
carbons
are
missing
between
the
5-‐membered
lactone
ring
and
the
chain
with
a
tertiary
butyl
group
in
butyrolactol
C
compared
to
butyrolactol
A.
The
full
genome
analysis
of
the
producer
enabled
us
to
identify
the
biosynthetic
gene
cluster
of
the
butyrolactols.
Further
analysis
indicated
that
the
t-‐butyl
starting
unit
could
be
originat-‐
ed
from
valine
catalysed
by
an
AdoCbl-‐dependent
isomerase
[3]
and
polyol
groups
could
be
synthesized
with
glycolate
extender
units.
Variation
of
the
chain
lengths
in
butyrolactols
A-‐D
will
be
discussed.
Relative
configuration
of
Butyrolactol
A
Acknowledgements:
This
work
was
supported
by
a
National
Research
Foundation
of
Korea
(NRF)
grant
funded
by
the
Korean
government
(Ministry
of
ICT
and
Future
Planning)
(2014R1A2A1A11053477)
and
a
grant
Marine
Biotechnology
Program
(Genome
Analysis
of
Marine
Organisms
and
Development
of
Functional
Applications,
PJT200620)
funded
by
Ministry
of
Oceans
and
Fisheries,
Korea.
Keywords: Butyrolactol, structure determination, microbial secondary metabolite, biosyn-‐
thesis
References:
[1] Bhatnagar
I,
Kim
S-‐K.
Pharmacologically
prospective
antibiotic
agents
and
their
sources:
A
marine
microbial
perspective.
Environ
Toxicol
Pharmacol
2012;
34:
631-‐643
[2] Kotake
C,
Yamasaki
T,
Moriyama
T,
Shinoda
M,
Komiyama
N,
Furumai
T,
Konishi
M,
Oki
T.
Butyrolactol
A
and,
B
new
antifungal
antibiotics
taxonomy,
isolation,
physici-‐chemical
properties,
structure
and
biological
activity.
J
Antibiot
1992;
45:
1442-‐1450
[3] Cracan
V,
Banerjee
R.
Novel
coenzyme
B12-‐dependent
interconversion
of
isovaler-‐CoA
and
p-‐CoA.
J
Biol
Chem
2012;
287:
3723-‐3732
P576
Stromatolite
microbial
communities
as
a
source
of
new
bioactive
secondary
metabolites
Patricia
M.
Flatt1,
Caro
Damarjanan2,
Eric
Isamonger2,
Jarmo
C.J.
Kalinski2,
Rosemary
A.
Dor-‐
rington2,
Kerry
L.
McPhail3
1Department
of
Chemistry,
Western
Oregon
University,
Monmouth,
OR
97361,
USA,
2Department
of
Bio-‐
chemistry
and
Microbiology,
Rhodes
University,
Grahamstown
South
Africa,
3Department
of
Pharmaceu-‐
tical
Sciences,
College
of
Pharmacy,
Oregon
State
University,
Corvallis,
OR
97331,
USA.
Stromatolites
represent
some
of
the
earliest
microbial
communities
on
Earth.
They
are
formed
by
accretion
and
precipitation
of
layered
calcium
carbonate
structures
that
result
from
the
metabolic
activity
of
complex
microbial
communities
and
the
geochemical
conditions
of
their
environment.
Modern
stromatolite
communities
include
aerobic
heterotrophs,
sul-‐
phide-‐oxidizing
bacteria,
sulphate-‐reducing
bacteria,
fermentative
bacteria
and
cyanobacte-‐
ria.
Phylogenetic
analyses
revealed
the
presence
of
new
and
known
cyanobacterial
taxa
relat-‐
ed
to
known
producers
of
biologically
active
secondary
metabolites
in
tufa
stromatolites
along
the
South
African
southeast
coast
[1].
Prompted
us
to
investigate
their
potential
for
producing
novel
bioactive
secondary
metabolites.
A
series
of
three
tide
pools
provided
the
opportunity
to
collect
stromatolites
along
a
vertical
transect
from
pool
A
(highest
elevation,
low
nitrogen
input,
fresh
water),
pool
B
(within
high
tide
zone,
brackish
water)
and
pool
C
(within
tidal
zone).
The
microbial
community
in
pool
A
is
particularly
distinct.
Chemical
ex-‐
tracts
of
stromatolites
from
different
pools
have
been
profiled
by
LC-‐MS/MS
and
the
data
sub-‐
jected
to
molecular
spectral
networking
using
the
GnPS
platform
[2]
in
order
to
establish
the
diversity
and
biological
potential
of
the
microbial
metabolome
that
is
being
expressed
within
each
of
these
microhabitats.
Correlation
of
the
phylogenetic
and
secondary
metabolomic
data
is
expected
to
guide
the
isolation
of
new
natural
products
with
biomedical
relevance.
Keywords: Stromatolites, cyanobacteria, LC-‐MS/MS, spectral networking
References:
[1] Perissinotto
R,
Bornman
TG,
Steyn
P-‐P,
Miranda
NAF,
Dorrington
RA,
Matcher
GF,
Strydom
N,
Peer
N.
Tufa
stromatolite
ecosystems
on
the
South
African
south
coast.
S
Afr
J
Sci
2014;
110:
1-‐8
[2] Guthals
A,
Watrous
JD,
Dorrestein
PC,
Bandeira
N.
The
spectral
networks
paradigm
in
high
throughput
mass
spectrometry.
Mol
Biosyst
2012;
8:
2535-‐2544
P577
Isolation,
structure
elucidation,
and
total
synthesis
of
an
N-‐
methylated
tetrapeptide
from
a
Panamanian
marine
cyanobacte-‐
rium
Kh.
Tanvir
Ahmed1,
Chakicherla
Gayathri3,
Laura
Pineda2,
Carmenza
Spadafora2,
Roberto
Gil3,
William
H.
Gerwick4,
Kevin
J.
Tidgewell1
1
Division
of
Pharmaceutical
Sciences,
Mylan
School
of
Pharmacy,
Duquesne
University,
600
Forbes
Ave.
Pittsburgh,
PA,
15282,
USA,
2
Instituto
de
Investigaciones
Científicas
y
Servicios
de
Alta
Tecnología
(IN-‐
DICASAT-‐AIP),
City
of
Knowledge
,
Apartado
0816-‐02852,
Panama
City,
Panama,
3
Department
of
Chem-‐
istry,
Carnegie
Mellon
University,
4400
Fifth
Avenue,
Pittsburgh,
PA,
15213,
USA,
4
Center
for
Marine
Bio-‐
technology
and
Biomedicine,
Scripps
Institution
of
Oceanography,
University
of
California
at
San
Diego,
La
Jolla,
CA
92037,
USA
As
part
of
the
Panama
ICBG
efforts
to
discover
novel
compounds
to
be
used
for
the
treatment
of
tropical
diseases
and
cancer,
a
cyanobacterial
extract
was
shown
to
have
activity
against
the
Trypanosoma
cruzi
and
Plasmodium
falciparum
parasites.
An
organic
extract
was
made
from
an
orange
cyanobacterium
collected
in
the
Portobelo
National
Park
on
the
Caribbean
side
of
Panama.
The
crude
extract
was
fractionated
into
nine
fractions
using
silica
gel
chroma-‐
tography
and
screened
for
activity
against
tropical
parasites
(T.
cruzi,
L.
donavani,
P.
falcipa-‐
rum)
and
cancer
cell
cytotoxicity.
One
of
these
subfractions
showed
75%
inhibition
of
growth
of
T.
cruzi
with
greater
activity
(97%)
against
P.
falciparum.
Further
purification
using
Sep-‐
Pak,
gave
several
fractions
with
selective
activity
against
malria
(>90%)
and
a
fraction
that
possessed
roughly
equipotent
~80%
inhibition
of
T.
cruzi
and
P.
falciparum.
The
malaria
se-‐
lective
fractions
were
shown
to
possess
a
known
compound
while
dereplication
of
the
other
fraction
showed
no
known
compounds
present.
HPLC
purification
resulted
in
identification
of
a
novel
N-‐methylated
tetrapeptide.
This
tetrapeptide
was
given
the
trivial
name
naranjamide
and
its
structure
determined
using
1-‐
and
2-‐D
NMR
and
LC-‐MS-‐MS.
To
confirm
configuration
and
activity,
a
total
synthesis
of
the
compound
is
being
undertaken.
In
addition
to
the
natural
product,
analogues
with
variable
N-‐methylation
are
being
synthesized
and
will
be
screened
for
activity.
Isolation,
structure
elucidation,
and
total
synthesis
of
naranjamide
have
been
un-‐
dertaken.
Acknowledgements:
The
authors
would
like
to
acknowledge
the
following
funding
sources:
ICBG
grant
(U01TW006634)
to
WHG
and
ASP
Research
Starter
Grant
to
KT.
We
would
also
like
to
acknowledge
the
Autoridad
Nacional
del
Ambiente
(ANAM)
and
Panama
for
allowing
us
to
collect
samples
in
Panama.
Keywords: Chagas' disease, malaria, marine natural product, total synthesis, cyanobacteria
P578
NMR
characterization
of
complex
natural
products:
Assigning
novel,
proton-‐deficient
alkaloid
scaffolds
Kirk R. Gustafson, Susanna T. S. Chan, Dennis Milanowski
Molecular Targets Laboratory, Center for Cancer Research, National Cancer Institute-‐Frederick, USA
NMR
provides
powerful
structural
elucidation
tools
that
are
particularly
well
suited
for
natu-‐
ral
products
studies.
Comprehensive
spectroscopic
characterization
of
a
native
metabolite
is
often
sufficient
to
fully
assign
a
new
structure.
However
assignment
of
novel
molecular
archi-‐
tectures
that
incorporate
numerous
heteroatoms
and
proton-‐deficient
fused
ring
systems
can
be
challenging.
Two
new
families
of
marine
alkaloids
were
recently
discovered
that
each
had
quite
novel
structural
features.
One
series
of
alkaloids
had
two
pyrimidine
rings
and
an
imid-‐
azole
ring
fused
to
generate
an
unprecedented
tetracyclic
core
with
embedded
guanidine
and
amidine
functionalities.
The
other
class
of
metabolites
incorporated
multiple
halogen
sub-‐
stituents
and
six
fused
heterocyclic
rings
to
generate
a
new
structural
motif.
Structure
char-‐
acterization
of
these
proton-‐deficient
alkaloid
scaffolds
was
quite
challenging
using
only
HMBC
heteronuclear
correlation
data,
which
generally
provides
only
two-‐
and
three-‐bond
correlations
with
good
sensitivity.
However,
application
of
some
new
NMR
pulse
sequences
(LR-‐HSQMBC,
HD-‐ADEQUATE)
and
strategies
ultimately
allowed
the
complete
structural
elu-‐
cidation
of
these
new
marine
alkaloids.
Continued
development,
refinement,
and
application
of
enhanced
NMR
capabilities
is
crucial
for
the
successful
assignment
of
novel
structural
scaf-‐
folds
and
functional
group
arrays
that
are
often
found
in
natural
products.
P579
Antileukemic
sesterterpenoids
from
a
marine
sponge,
Luffariella
sp.
Kuei-‐Hung
Lai1,2,
Mei-‐Chin
Lu3,4,
Fang-‐Rong
Chang2,5,6,
Jui-‐Hsin
Su3,4,
Mohamed
El-‐Shazly2,7,
Ying-‐Chi
Du2,
Tung-‐Ying
Wu2,
Yu-‐Ming
Hsu2,
Anders
Backlund1,
Yang-‐Chang
Wu,2,8,9,10
1
Division
of
Pharmacognosy,
Department
of
Medicinal
Chemistry,
Uppsala
University,
Uppsala,
Sweden,
2
Graduate
Institute
of
Natural
Products,
College
of
Pharmacy,
Kaohsiung
Medical
University,
Kaohsiung
807,
Taiwan,
3
Graduate
Institute
of
Marine
Biotechnology,
National
Dong
Hwa
University,
Pingtung
944,
Taiwan,
4
National
Museum
of
Marine
Biology
and
Aquarium,
Pingtung
944,
Taiwan,
5
Cancer
Center,
Kaohsiung
Medical
University
Hospital,
Kaohsiung
80708,
Taiwan,
6
Research
Center
for
Natural
Product
and
New
Drug,
Kaohsiung
Medical
University,
Kaohsiung
80708,
Taiwan,
7
Department
of
Pharmacogno-‐
sy
and
Natural
Products
Chemistry,
Faculty
of
Pharmacy,
Ain-‐Shams
University,
Organization
of
African
Unity
Street,
Abassia,
Cairo
11566,
Egypt,
8
School
of
Pharmacy,
College
of
Pharmacy,
China
Medical
Uni-‐
versity,
Taichung
40402,
Taiwan,
9
Chinese
Medicine
Research
and
Development
Center,
China
Medical
University
Hospital,
Taichung
40447,
Taiwan,
10
Center
for
Molecular
Medicine,
China
Medical
University
Hospital,
Taichung
40447,
Taiwan
Marine
sponge
of
the
genus
Luffariella
was
found
to
be
a
rich
source
of
novel
cytotoxic
and
anti-‐inflammatory
sesterterpenoids
from
previous
reports
[1,
2].
The
further
chemical
inves-‐
tigation
on
this
sponge
led
to
the
discovery
of
five
new
sesterterpenoids,
luffarlides
G–K,
which
were
elucidated
by
spectroscopic
methods
including
1D
and
2D
NMR
techniques.
The
absolute
configurations
were
determined
utilizing
ECD/CD
experiments.
The
antileukemic
activity
of
the
new
compounds,
along
with
the
six
previously
described
sesterterpenoids,
were
evaluated
against
Molt4
human
acute
lymphoblastic
leukemic
cell
lines
using
MTT
pro-‐
liferative
assay
[3].
The
most
active
compound,
manoalide,
was
found
to
exhibit
strong
anti-‐
proliferative
activitie
with
IC50
values
of
0.14
μg/mL.
In
addition,
manoalide
treatment
acti-‐
vated
caspase-‐related
apoptotic
proteins
and
mitogen-‐activated
protein
kinases
(MAPK)
of
Molt4
cells
with
Western
blot
assay
[3].
Analyzed
by
flow
cytometry,
manoalide
treatment
induced
cancer
cells
apoptosis
via
disruption
of
mitochondrial
membrane
potential
and
gen-‐
eration
of
intracellular
reactive
oxygen
species
(ROS)
in
Molt4
cells.
Manoalide
could
inhibit
both
activities
of
human
topoisomerase
I
and
II
with
gel
electrophoresis
[3].
DNA
damage
re-‐
sponses
were
directly
detected
with
increase
of
H2AX
phosphorylation,
biomarker
of
DNA
damage,
and
DNA
break
by
comet
assay.
Importantly,
the
apoptosis-‐caused
by
manoalide
could
be
recovered
with
NAC
pretreatment,
but
not
with
pretreatments
of
caspase
and
MAPK
inhibitors.
Taken
together,
these
results
suggested
that
cytotoxic
effect
of
manoalide
is
to
me-‐
diate
ROS
generation,
and
finally
resulted
in
cellular
DNA
damage
and
apoptosis.
The
findings
provide
support
for
further
ChemGPS-‐NP
targets
investigation
as
well
as
the
in
vivo
evaluation
of
lead
compound.
Keywords:
Luffarilla
sp.,
sesterterpenoids,
manoalide,
Molt4,
ROS-‐mediated
apoptosis,
DNA
damage
References:
[1] Kernan
MR,
Faulkner
DJ,
Parkanyi
L,
Clardy
J,
Decarvalho
MS,
Jacobs
RS.
Luffolide,
a
novel
anti-‐Inflammatory
terpene
from
the
sponge
Luffariella
sp.
Experientia
1989;
45:
388-‐390
[2] Zhou
GX,
Molinski
TF.
Manoalide
derivatives
from
a
sponge,
Luffariella
sp.
J
Asian
Nat
Prod
Res,
2006;
8:
15-‐20
[3] Shih
SP,
Lee
MG,
El-‐Shazly
M,
Juan
YS,
Wen
ZH,
Du
YC,
Su
JH,
Sung
PJ,
Chen
YC,
Yang
JC,
Wu
YC,
Lu
MC.
Tackling
the
cytotoxic
effect
of
a
marine
polycyclic
quinone-‐type
metabolite:
Halenaquinone
induces
molt
4
cells
apoptosis
via
oxidative
stress
combined
with
the
inhibition
of
HDAC
and
topoisomerase
activities.
Mar
Drugs
2015;
13:
3132-‐3153
P580
Isolation
and
identification
of
new
secondary
metabolites
from
the
marine
sponge
Monanchora
unguiculata
Pierre-‐Eric Campos1, Emerson Ferreira Queiroz2, Laurence Marcourt2, Jean-‐Luc Wolfender2,
Anne-‐Sophie Sanchez1, Bertrand Illien1, Ali Al Mourabit3, Anne Gauvin-‐Bialecki1
1Laboratoire de Chimie des Substances Naturelles et des Sciences des Aliments, Université de la Réunion,
15
Avenue
René
Cassin,
CS
92003,
97744
Saint
Denis
CEDEX
9,
France,
2School
of
pharmaceutical
scienc-‐
es,
University
of
Geneva,
University
of
Lausanne,
30
Quai
Ernest-‐Ansermet,
1211
Geneva
4,
Switzerland,
3Centre
de
Recherche
de
Gif-‐sur-‐Yvette,
Institut
de
Chimie
des
Substances
Naturelles,
UPR
2301,
CNRS,
Avenue de la Terrasse, 91198 Gif-‐sur-‐Yvette, France
Marine
sponges
naturally
produce
toxic
molecules
that
prevent
other
organisms
from
exploit-‐
ing
them.
Some
of
these
substances
are
also
known
for
their
therapeutic
properties
such
as
Ara-‐A
and
Ara-‐C,
two
analogues
with
relevant
antiviral
activity
or
more
recently
bryostatin
1
or
dehydrodidemnin
B
[1].
In
our
continuous
search
for
bioactive
metabolites
from
marine
invertebrates,
the
Madagas-‐
car
sponge
Monanchora
unguiculata
has
been
investigated.
Its
crude
extract
was
found
to
be
cytotoxic
on
KB
cells
(99%
at
10
mg/mL)
and
active
against
the
malaria
parasite
Plasmodium
falciparum
(IC50
<
5
mg/mL
at
2.26
mg/mL).
The
chemical
investigation
of
the
crude
extract
led
to
the
isolation
of
eight
compounds
(1-‐8)
including
four
new
guanidine
alkaloids
(5-‐8).
The
four
known
compounds
are
crambescidin
800
(1),
crambescidin
359
(2),
crambescidic
acid
(3)
and
fromiamycalin
(4)
previously
isolated
from
Crambe
crambe,
Monanchora
unguic-‐
ulata,
Monanchora
unguifera
and
Fromia
monilis
respectively.
Crambescidin
800
is
already
known
for
having
some
biological
activities
such
as
anti-‐HIV-‐1
or
antimalarial
activities
[2,3].
The
four
new
compounds
(5-‐8)
were
characterized
by
HRMS
and
NMR.
However,
for
com-‐
pounds
7
and
8
more
chemical
investigations
are
in
progress
in
order
to
confirm
their
struc-‐
tural
elucidation.
Bioassays
will
be
conducted
in
order
to
evaluate
the
cytotoxicity
on
KB
cells
and
the
antimalarial
activity
of
the
isolated
compounds.
Compound (5)
Monanchora unguiculata Compound (6)
Keywords:
Marine
natural
products,
Monanchora,
Crambescidins,
Alkaloids.
References:
[1] Kijjoa
A,
Sawangwong
P.Drugs
and
Cosmetics
from
the
Sea.
Marine
Drugs
2004;
2:
73–82
[2] Hua
HM,
Peng
J,
Dunbar
DC,
Schinazi
RF,
Andrews
AGDC,
Cuevas
C,
Garcia-‐Fernandez
LF,
Kelly
M,
Hamann
MT.
Batzelladine
Alkaloids
from
the
Caribbean
Sponge
Monanchora
un-‐
guifera
and
the
Significant
Activities
against
HIV-‐1
and
AIDS
Opportunistic
Infectious
Pathogens,
Tetrahedron
2007;
45:
11179–11188
[3] Chang
N,
Whittaker
F,
Bewley
CA.
Crambescidin
826
and
Dehydrocrambine
A:
New
Poly-‐
cyclic
Guanidine
Alkaloids
from
the
Marine
Sponge
Monanchora
Sp.
That
Inhibit
HIV-‐1
Fusion,
J
Nat
Prod
2003;
11:
1490–1494
P581
Induction
of
cryptic
metabolites
from
a
rare
Antarctic
psychro-‐
phile,
Marinobacter
sp.
Jeremy
Carter1,
Miller
Judge1,
Lauren
McLean1,
Jill
Mikucki2,
Lesley-‐Ann
Giddings1
1
Department
of
Chemistry
&
Biochemistry,
Middlebury
College,
276
Bicentennial
Way,
05753,
Middle-‐
bury, USA, 2 Department & Biology, Middlebury College, 276 Bicentennial Way, 05753, Middlebury, USA
Microbes from extreme environments represent an untapped source of novel, bioactive me-‐
tabolites with unique molecular frameworks, as they have evolved genes that are critical for
surviving in unusual environments [1-‐3]. Antarctic subglacial environments are among the
most challenging systems to study because they are locked under 100–1000s of meters of gla-‐
cial ice and require novel drilling technology to access materials cleanly for microbiological
analyses. Only recently have microbiological samples been obtained from these cold, dark
isolated ecosystems. Blood Falls, one of the better-‐characterized subglacial environments in
Antarctica, is a cold (-‐7 °C), iron-‐rich (~3.4 mM) subglacial brine (8% NaCl) that leaks out
from below the Taylor Glacier, Antarctica [4]. We recently isolated and characterized a mod-‐
erately halophilic, heterotrophic psychrophile from the Blood Falls brine that clusters within
the Marinobacter genus. Bioinformatic analysis of this strain’s genome indicated the presence
of at least four gene clusters involved in secondary metabolism with low sequence identity
(~30%) to other known genes in GenBank. Two gene clusters are most similar to those that
produce aryl polyenes, which function as pigments/antioxidants, protecting bacteria from
reactive oxygen species. Another gene cluster appears to be involved in terpene biosynthesis
and most likely produces pigments, as it contains includes lycopene cyclase from carotenoid
biosynthesis. To identify some of these uncharacterized, cryptic gene products with potential
bioactivity, we modified growth conditions as well as created a cosmid library in Escherichia
coli to induce the production of pigments. We were able to induce gene expression and par-‐
tially characterize metabolic products by high-‐resolution LC/MS by varying media as shown
in the figure below. These data provide insight into the metabolome of Marinobacter sp., and
its role in cold environments, such as those found in Antarctica.
NL:
RT: 0.00000 - 10.00770 SM: 15B 1.44E6
100 431.2763 413.2662 Marine broth TIC MS
lg32p1_01
95 NL:
90 TCG medium 2.70E6
TIC MS
85 lg32p4_35
NL:
80 Glucose peptone 1.24E6
75 yeast extract TIC MS
LG32P5_03
Relative Abundance
70
65
60
55 245.1284 413.2662
50
45 488.2963 415.2817
40
35 467.2058
415.2817
30 211.1441
431.2767
25 211.1441
20 301.1410 413.2662
15 211.1441 413.2662 413.2662 413.2662
413.2662 413.2662
10 337.1562 413.2662
413.2662
5
0
0 1 2 3 4 5 6 7 8 9 10
Photo credit: Peter Rejcek
Time (min)
Acknowledgements:
We
would
like
to
acknowledge
Professor
Russell
Kerr
at
the
University
of
Prince
Ed-‐
ward
Island
for
kindly
providing
access
to
his
high-‐resolution
LC-‐MS.
Keywords: Cryptic metabolites, extremophilic microbes, bioactive compounds
References:
[1] Giddings
L-‐A,
Newman
DJ.
Bioactive
compounds
from
extremophiles:
genomic
studies,
biosynthetic
gene
clusters,
and
new
dereplication
methods.
In:
Springer;
2015
[2] Giddings
L-‐A,
Newman
DJ.
Bioactive
compounds
from
terrestrial
extremophiles.
In:
Springer;
2015
[3] Giddings
L-‐A,
Newman
DJ.
Bioactive
compounds
from
marine
extremophiles.
In:
Springer;
2015
[4] Mikucki
JA,
Pearson
A,
Johnston
DJ,
Turchyn
AV,
Farquhar
J,
Schrag
DP,
Anbar
AD,
Priscu
JC,
Lee
PA.
A
Contemporary
Microbially
Maintained
Subglacial
Ferrous
"Ocean".
Science
2009;
324:
397-‐400
P582
Biological
and
chemical
analysis
of
actinomycetes
associated
with
marine
sponges
from
Singapore
Gary Chi Ying Ding, Lik Tong Tan
Natural
Sciences
and
Science
Education,
National
Institute
of
Education,
1
Nanyang
Walk,
Singapore
637616
Marine
invertebrates,
such
as
sponges
and
corals,
are
hosts
to
a
myriad
of
microbes
and
they
represent
potential
sources
of
useful
bioactive
natural
products
[1].
In
this
study,
we
present
biological
and
chemical
data
of
actinomycete
strains
isolated
from
common
marine
sponges,
including
Gelliodes
fibulata,
Clathria
reinwardti
and
Xestospongia
testudinaria,
found
in
Singa-‐
pore
reefs.
Using
culture-‐dependent
method,
these
marine
microbes
were
isolated
and
cul-‐
tured
based
on
11
different
marine
media.
These
strains
were
further
selected
based
on
ge-‐
nomic
profiling
using
antiSMASH
for
the
presence
of
novel
polyketide
synthase
and
non-‐
ribosomal
peptide
synthetase
gene
clusters
[2].
In
addition,
the
organic
extracts
of
selected
marine
microbes
were
screened
for
cytotoxicity
and
antibacterial
activity
based
on
Escherich-‐
ia
coli
and
Bacillus
cereus.
The
phylogenetic
data
based
on
16S
rRNA
genes
of
selective
marine
bacterial
strains
that
showed
significant
biological
activities
is
also
presented.
In
addition,
preliminary
chemical
data
based
on
LC-‐MS
and
1H-‐NMR
spectroscopy
of
bioactive
bacterial-‐
derived
extracts/fractions
indicated
the
presence
of
unique
secondary
metabolites.
This
study
revealed
the
biotechnological
applications
of
local
marine
invertebrate-‐associated
microbes
in
drug
discovery
and
development
efforts.
Acknowledgements:
The
authors
would
like
to
acknowledge
the
NIE
AcRF
(RI
2/14
TLT)
for
financial
support.
Keywords: Marine biotechnology, drug discovery, sponge-‐associated actinomycetes, bioac-‐
tive natural products
References:
[1] Leal
MC,
Sheridan
C,
Osinga
R,
Dionisio
G,
Rocha
RJ,
Silva
B,
Rosa
R,
Calado
R.
Marine
mi-‐
croorganism-‐invertebrate
assemblages:
perspective
to
solve
the
"supply
problem"
in
the
initial
steps
of
drug
discovery.
Mar
Drugs
2014;
12:
3929–3952.
[2] Weber
T,
Blin
K,
Duddela
S,
Krug
D,
Kim
HU,
Bruccoleri
R,
Lee
SY,
Fischbach
MA,
Muller
R,
Wohlleben
W,
Breitling
R,
Takano
E,
Medema
MH.
antiSMASH
3.0
-‐
a
comprehensive
re-‐
source
for
the
genome
mining
of
biosynthetic
gene
clusters.
Nucleic
Acids
Res
2015;
43:
W237–243.
P583
Chemical
constituents
of
the
soft
coral
Sinularia
gravis
from
the
coast
of
Madagascar
Marie
Pascaline
Rahelivao,
Margit
Gruner,
Hans-‐Joachim
Knölker
Technische
Universität
Dresden,
Department
Chemie,
Bergstrasse
66,
01069
Dresden,
Germany
Secondary
metabolites
obtained
from
marine
organisms
including
animals,
plants,
and
micro-‐
organisms
are
a
rich
source
for
new
pharmacologically
active
compounds
[1].
The
sea
waters
of
Madagascar,
the
fourth
largest
island
in
the
world
with
nearly
5000
km
of
coastline,
ac-‐
commodates
a
wealth
of
marine
organisms
in
its
coastal
region.
To
date,
only
few
representa-‐
tives
of
Madagascan
soft
corals
have
been
investigated
for
their
chemical
constituents
[2−4].
The
present
study
aims
at
the
investigation
of
the
chemical
constituents
of
the
soft
coral
Sinu-‐
laria
gravis
collected
at
the
coast
of
Madagascar.
Four
new
compounds,
the
spatane-‐type
diterpenoid
gravilin
(1),
the
monoalkylmonoacylglycerol
2,
the
dihomoditerpenoid
ketone
3,
and
isodecaryiol
(4),
along
with
the
three
known
compounds
(+)-‐(S)-‐geranyllinalool,
(−)-‐(R)-‐
nephthenol,
and
11,12-‐epoxysarcophytol
A
have
been
isolated
from
S.
gravis
[5].
The
isolated
compounds
were
characterized
using
1H
NMR
and
13C
NMR
spectroscopy.
In
combination
with
COSY,
NOESY,
HMBC,
and
HSQC
measurements,
a
complete
structural
elucidation
was
achieved.
The
structure
of
compound
4
including
its
absolute
stereochemistry
was
confirmed
by
single
crystal
X-‐ray
diffraction.
Acknowledgements:
Marie
Pascaline
Rahelivao
is
grateful
to
the
European
Commission
(Erasmus
Mundus
Programme),
the
Gesellschaft
von
Freunden
und
Förderern
der
TU
Dresden
(GFF
TU
Dresden),
and
the
TUD
Graduate
Academy
for
their
support
by
providing
scholarships.
Keywords: Soft corals, Cembranoid, Spatane diterpenes, NMR spectroscopy, X-‐ray diffraction
References:
[1] Jha
RK,
Zi-‐rong
X.
Biomedical
compounds
from
marine
organisms.
Mar
Drugs
2004;
2:
123–146
[2] Poza
JJ,
Fernández
R,
Reyes
F,
Rodríguez
J,
Jiménez
C.
Isolation,
biological
significance,
syn-‐
thesis,
and
cytotoxic
evaluation
of
new
natural
parathiosteroids
A−C
and
analogues
from
the
soft
coral
Paragorgia
sp.
J
Org
Chem
2008;
73:
7978–7984
[3] Longeon
A,
Bourguet-‐Kondracki
M-‐L,
Guyot
M.
Two
new
cembrane
diterpenes
from
a
Madagascan
soft
coral
of
the
genus
Sarcophyton.
Tetrahedron
Lett
2002;
43:
5937–5939
[4] Hou
Y,
Harinantenaina
L.
New
and
bioactive
natural
products
isolated
from
Madagascar
plants
and
marine
organisms.
Curr
Med
Chem
2010;
17:
1191–1219
[5] Rahelivao
MP,
Gruner
M,
Lübken
T,
Islamov
D,
Kataeva
O,
Andriamanantoanina
H,
Bauer
I,
Knölker
H-‐J.
Chemical
constituents
of
the
soft
corals
Sinularia
vanderlandi
and
Sinularia
gravis
from
the
coast
of
Madagascar.
Org
Biomol
Chem
2016;
14:
989–1001
P584
Antibiotic
tetramic
acids
from
marine
fungal
endophytes
cul-‐
tured
under
epigenetic
regulation
Matthew
A.
Knestrick1,
Renee
Fleeman2,
Lindsey
N.
Shaw2,
Bill
J.
Baker1
Departments
of
1
Chemistry,
2
Cell
Biology,
Microbiology,
and
Molecular
Biology,
and
the
Center
for
Drug
Discovery
and
Innovation,
University
of
South
Florida,
Tampa,
FL
33620,
USA
The
ESKAPE
pathogens
represent
a
group
of
common
and
deadly
drug
resistant,
bacterial
pathogens.
Among
the
ESKAPE
pathogens,
methycilin-‐resistant
Staphylococcus
aureus
(MRSA)
is
responsible
for
more
fatalities
in
US
hospitals
than
HIV/AIDS
and
tuberculosis
combined
[1].
In
the
face
of
such
increasingly
resistant
pathogens,
there
is
a
dire
need
for
new
and
novel
drug
candidates.
Our
search
for
new
chemodiversity
in
support
of
an
antibiotic
drug
disovery
screening
pro-‐
gram
focuses
on
niche
biodiversity.
One
such
niche
environment
is
the
coastal
and
marine-‐
margine
community.
Mangroves
and
their
associated
fauna
are
rich
with
endophytic
fungi.
Our
mangrove
endophyte
collection
has
been
cultivated
with
and
without
epigenetic
regula-‐
tion
with
the
aim
of
eliciting
latent
secondary
metabolite
pathways.
A
fungal
strain,
KML14-‐75MG,
isolated
from
the
mangrove
plant
Pandanus
spiralis
was
identi-‐
fied
for
its
activity
against
MRSA.
KML14-‐75MG
was
cultured
on
nutirent-‐enriched
rice
treat-‐
ed
with
5-‐azacytidine,
a
DNA
methyltransferase
(DNMT)
inhibitor
known
to
upregulate
some
secondary
metabolite
pathways
[2].
The
extract
was
purified
using
a
bioassay
guided
fractionation
approach,
leading
to
a
suite
of
related
tetramic
acids
with
potent
activity
against
MRSA.
Structure
elucidation
of
these
tetramic
acids
was
conducting
using
both
1D
and
2D
Nuclear
Magnetic
Resonance
(NMR)
Spectroscopy,
with
molecular
formula
and
mass
determined
by
Liquid
Chromatography
hy-‐
brid
Quadrupole
Time-‐of-‐Flight
High
Resolution
Mass
Spectrometry
(LC-‐QtoF
HRMS).
Both
new
and
known
tetramic
acids
were
identified,
with
epigenetic
regulation
eliciting
the
pro-‐
duction
of
new
compounds
not
present
in
the
untreated
fungal
strain.
Acknowledgments:
We
are
grateful
for
financial
support
from
the
NIH
(AI103715),
as
well
the
State
of
Florida
for
Center
of
Excellence
funding
of
CDDI.
Keywords:
ESKAPE
pathogens,
epigenetics,
drug
discovery
References:
[3] Boucher
HW,
Talbot
GH,
Bradley
JS,
Edwards
JE,
Gilbert
D,
Rice
LB,
Scheld
M,
Spellberg
B,
Bartlett
B.
Bad
bugs,
no
drugs:
no
ESKAPE!
An
update
from
the
Infectious
Diseases
Society
of
America.
Clin
Infect
Dis
2015;
48:
1-‐12
[4] Fisch
KM,
Gillaspy
AF,
Gipson
M,
Henrikson
JC,
Hoover
AR,
Jackson
L,
Najar
FZ,
Wagele
H,
Cichewicz
RH.
Chemical
induction
of
silent
biosynthetic
pathway
transcription
in
Aspergillus
niger.
J
Ind
Microbiol
Biotechnol
2009;
36:
1199-‐1213
P585
Alpha-‐glucosidase
and
acetylcholinesterase
inhibitory
activities
of
phlorotannins
isolated
from
the
brown
alga,
Ecklonia
maxima
Mutalib
A.
Aderogba1,2,
Rengasamy
R.R.
Kannan,
Ashwell
R.
Ndhlala2,
Johannes
Van
Staden2
1
Department
of
Chemistry,
Obafemi
Awolowo
University,
Ile-‐Ife
220282,
Nigeria,
2
Research
Centre
for
Plant
Growth
and
Development,
School
of
Life
Sciences,
University
of
KwaZulu-‐Natal
Pietermaritzburg,
Scottsville
3209,
South
Africa
Phytochemical
investigation
of
plants
for
bioactive
constituents
offers
a
great
opportunity
for
discovery
of
novel
therapeutic
agents.
It
also
provides
useful
lead
compounds
for
structural
modification
to
obtain
better
biological
activity
and
moderate
side
effects.
Many
drugs
that
are
in
use
in
modern
medicine
have
their
origin
from
plants.
Ecklonia
maxima
(Osbeck)
Pa-‐
penfuss.,
a
brown
alga
from
the
west
coast
of
South
Africa
was
investigated
for
its
bioactive
constituents
for
the
management
of
neurodegenerative
and
diabetes
disorders
[1,
2].
The
crude
extract,
solvent
fractions
and
phlorotannins
isolated
from
a
80%
methanol
E.
maxima
extract
were
evaluated
for
α-‐glucosidase
and
acetylcholinesterase
(AChE)
inhibitory
activities
using
microplate
techniques.
The
IC50
values
for
the
solvent
fractions
in
the
α-‐glucosidase
as-‐
say
ranged
from
3.91
to
31.14
μg/mL,
while
in
the
AChE
assay,
the
IC50
values
for
the
solvent
fractions
ranged
from
62.61
to
150.8
μg/mL,
with
the
ethyl
acetate
fraction
having
the
best
inhibitory
activity
against
AChE.
Repeated
column
fractionation
of
the
ethyl
acetate
fraction
on
Sephadex
LH-‐20
resulted
in
isolation
of
three
compounds.
Structure
elucidation
of
the
iso-‐
lated
compounds
was
carried
out
using
spectroscopic
techniques:
mass
spectrometry
(ESI-‐
TOF-‐MS)
and
NMR
(1D
and
2D
experiments).
These
compounds
from
E.
maxima
were
identi-‐
fied
as:
1,3,5-‐trihydroxybenzene
(phloroglucinol)
(1),
together
with
two
of
its
derivatives
dibenzo
[1,4]
dioxine-‐2,4,7,9-‐tetraol
(2)
and
hexahydroxyphenoxydibenzo
[1,4]
dioxine
(eck-‐
ol)
(3).
The
spectra
data
of
compound
2
is
reported
for
the
first
time.
In
the
α-‐glucosidase
as-‐
say,
the
IC50
values
for
compounds
(2)
and
(3)
were
33.2
and
11.2
μM
respectively,
more
po-‐
tent
than
the
standard
acarbose
(1013.4
μM).
AChE
inhibitory
activity
of
the
compounds
ranged
from
IC50
=
76.7
to
579.3
μM,
with
compounds
2
and
3
having
the
best
activity,
lower
than
the
IC50
of
the
standard
galanthamine
(1.8
±
0.5
μM).
To
the
best
of
our
knowledge,
there
is
no
previous
report
on
antidiabetic
properties
of
phlorotannins
from
E.
maxima.
The
activi-‐
ties
demonstrated
by
the
extracts
and
the
isolated
compounds
in
these
studies
suggest
that
these
extracts
and
compounds
could
be
explored
as
novel
therapeutic
agents.
OH
OH OH
OH O OH
O OH
O OH
HO OH HO O
1 HO O
2 OH 3 OH
Acknowledgement:
The
authors
thank
the
University
of
KwaZulu-‐Natal
for
Postdoctoral
Fellowships.
Keywords: Ecklonia maxima, phlorotannins, alpha-‐glucosidase, diabetes mellitus, acetylcho-‐
linesterase
References:
[1] Kannan
RRR,
Aderogba
MA,
Ndhlala
AR,
Stirk
WA,
Van
Staden
J.
Acetylcholinesterase
in-‐
hibitory
activity
of
phlorotannins
isolated
from
the
brown
alga,
Ecklonia
maxi-‐
ma
(Osbeck)
Papenfuss.
Food
Res
Int
2013,
54:1250-‐1254
[2] Kannan
RRR,
Aderogba
MA,
Amoo
SO,
Stirk
WA,
Van
Staden
J.
Potential
antiradical
and
al-‐
pha-‐glucosidase
inhibitors
from
Ecklonia
maxima
(Osbeck)
Papenfuss.
Food
Chem
2013;
141:
1412-‐1415
P586
Agelamadins,
bromopyrrole
alkaloids
from
Okinawan
marine
sponges
Agelas
spp.
Naonobu
Tanaka1,2,
Taishi
Kusama2,
Yoshiki
Kashiwada1,
Jun’ichi
Kobayashi2
1Graduate
School
of
Pharmaceutical
Sciences,
Tokushima
University,
Tokushima
770-‐8505,
Japan,
2Graduate
School
of
Pharmaceutical
Sciences,
Hokkaido
University,
Sapporo
060-‐0812,
Japan
Marine
sponges
belonging
to
the
genus
Agelas
are
recognized
as
a
rich
source
of
bro-‐
mopyrrole
alkaloids,
which
have
attracted
widespread
interest
due
to
their
intriguing
chemi-‐
cal
structures
and
biological
activities.
In
the
course
of
our
continuing
study
for
interesting
secondary
metabolites
from
marine
organisms,
two
marine
sponges
Agelas
spp.
collected
at
Okinawa
were
investigated
to
give
six
bromopyrrole
alkaloids,
agelamadins
A–F
(1–6).
The
structures
of
1–6
were
assigned
on
the
basis
of
spectroscopic
analyses.
Br
NH
NH 2 Br
HN O
H NH 2
NH Br HN Br
O Br N
NH HN NH
R H NH 2
9 H
HN N NH 3 O HN
10 N Br N
HN H N
H N
H 2N O O
Br N N HO 2C S
Br H H HH O
HO
agelamadin)C)(3)) 9R,)10S
agelamadin)A)(1) R)=)OMe agelamadin)F)(6)
agelamadin)D)(4) 9S,)10R
agelamadin)B)(2) R)=)OH
agelamadin)E)(5) 9R,)10R
Agelamadins
A
(1)
and
B
(2)
[1]
are
dimeric
bromopyrrole
alkaloids
consisting
of
an
agelas-‐
tatin-‐like
tetracyclic
moiety
and
an
oloidin-‐like
linear
moiety
in
common.
Agelamadins
C–E
(3–5)
[2]
were
assigned
as
conjugates
of
monomeric
bromopyrrole
alkaloid
and
hy-‐
droxykynurenine.
Their
absolute
configurations
were
elucidated
by
application
of
the
PGME
method
and
the
TDDFT
ECD
calculation.
Agelamadin
F
(5)
[3]
is
a
monomeric
bromopyrrole
alkaloid
comprising
oroidin
and
3-‐hydroxypyridinum.
The
isolation,
structure
elucidation,
and
antimicrobial
activity
of
these
alkaloids
will
be
presented.
Acknowledgements:
We
thank
Mr.
Z.
Nagahama
for
his
help
with
sponge
collection,
Dr.
J.
Fromont
for
sponge
identification,
and
Prof.
T.
Gonoi
for
antimicrobial
evaluation.
Keywords: Marine sponges, Agelas, bromopyrrole alkaloids, agelamadins
References:
[1] Kusama
T,
Tanaka
N,
Sakai
K,
Gonoi
T,
Fromont
J,
Kashiwada
Y,
Kobayashi
J.
Agelamadins
A
and
B,
dimeric
bromopyrrole
alkaloids
from
a
marine
sponge
Agelas
sp.
Org
Lett
2014;
16:
3916−3918
[2] Kusama
T,
Tanaka
N,
Sakai
K,
Gonoi
T,
Fromont
J,
Kashiwada
Y,
Kobayashi
J.
Agelamadins
C–E,
bromopyrrole
alkaloids
comprising
oroidin
and
3-‐hydroxykynurenine
from
a
marine
sponge
Agelas
sp.
Org
Lett
2014;
16:
5716−5719
[3] Kusama
T,
Tanaka
N,
Kashiwada
Y,
Kobayashi
J.
Agelamadin
F
and
tauroacidin
E,
bro-‐
mopyrrole
alkaloids
from
an
Okinawan
marine
sponge
Agelas
sp.
Tetrahedron
Lett
2015;
56:
4502−4504
P587
Bromotyrosine
alkaloids
with
acetylcholinesterase
inhibitory
ac-‐
tivity
from
the
Thai
sponge
Acanthodendrilla
sp.
Natchanun Sirimangkalakitti1, Masashi Yokoya2, Kanokwan Changwichit3, Tongchai Saesong3,
Opeyemi J. Olatunji4, Supakarn Chamni1, Pithi Chanvorachote5, Kornkanok Ingkaninan3,
Anuchit Plubrukarn4, Naoki Saito2, Khanit Suwanborirux1
1 Center for Bioactive Natural Products from Marine Organisms and Endophytic Fungi (BNPME), De-‐
partment
of
Pharmacognosy
and
Pharmaceutical
Botany,
Faculty
of
Pharmaceutical
Sciences,
Chulalongkorn
University,
Bangkok
10330,
Thailand,
2
Graduate
School
of
Pharmaceutical
Sciences,
Meiji
Pharmaceutical
University,
Tokyo
204–8588,
Japan,
3
Department
of
Pharmaceutical
Chemistry
and
Pharmacognosy,
Faculty
of
Pharmaceutical
Sciences
and
Center
of
Excellence
for
Innovation
in
Chemis-‐
try,
Naresuan
University,
Phitsanulok
65000,
Thailand,
4
Department
of
Pharmacognosy
and
Pharmaceu-‐
tical
Botany,
Faculty
of
Pharmaceutical
Sciences,
Prince
of
Songkla
University,
Hat-‐Yai,
Songkhla
90112,
Thailand,
5
Cell-‐Based
Drug
and
Health
Product
Development
Research
Unit
and
Department
of
Pharma-‐
cology
and
Physiology,
Faculty
of
Pharmaceutical
Sciences,
Chulalongkorn
University,
Bangkok
10330,
Thailand
Twenty-‐one bromotyrosine alkaloids, including two new compounds, 13-‐oxosubereamolline
D (5) and acanthodendrilline (21), were isolated from the MeOH crude extract of the Thai
sponge Acanthodendrilla sp. collected from Koh-‐Ha Island, Krabi, Thailand [1, 2]. Their struc-‐
tures were determined by several spectroscopic techniques involving 1D-‐ and 2D-‐NMR, high-‐
resolution mass, in conjugation with optical rotation and circular dichroism data. Importantly,
our result is the first report of the discovery of bromotyrosine alkaloids from the sponge be-‐
longing to the order Dendroceratida [1]. In addition, the S and R synthetic enantiomers of 21
were prepared to resolve the absolute configuration of its single chiral carbon at oxazolidone
moiety. Both enantiomers of 21 were obtained from the 4-‐step synthetic strategy using tyra-‐
mine as a precursor [2].
All
isolated
compounds
were
evaluated
for
the
inhibitory
activities
against
electric
eel
acetyl-‐
cholinesterase
(EeAChE)
and
human
recombinant
acetylcholinesterase
(hrAChE)
by
the
modi-‐
fied
Ellman’s
method
[3].
The
results
indicated
that
homoaerothionin
(7)
exhibited
the
strongest
inhibitory
activity
with
IC50s
of
2.9
and
4.5
μM
against
EeAChE
and
hrAChE,
respec-‐
tively.
The
hrAChE
inhibition
kinetics
of
7
showed
increased
Km
and
unchanged
Vmax
values,
suggesting
its
competitive
mode
of
inhibition.
The
spirocyclohexadienylisoxazole
and
specific
length
of
the
alkyl
diamine
linkage
were
proposed
as
the
crucial
parts
for
its
strong
inhibitory
activity.
This
finding
indicates
a
therapeutic
potential
for
7
in
acetylcholine-‐related
diseases,
most
importantly
Alzheimer's
disease.
Acknowledgements:
The
Thailand
Research
Fund
through
the
Royal
Golden
Jubilee
Ph.D.
Program
(Grant
No.
PHD/0276/2552)
is
acknowledged
for
financial
support.
Keywords: Bromotyrosine alkaloids, acetylcholinesterase inhibitor, sponge, Acanthodendrilla
References:
[1]
Sirimangkalakitti
N,
Olatunji
O,
Changwichit
K,
Saesong
T,
Chamni
S,
Chanvorachote
P,
Ingkaninan
K,
Plubrukarn
A.
Suwanborirux
K.
Bromotyrosine
marine
alkaloids
with
acetylcholinesterase
inhibitory
activity
from
the
Thai
sponge
Acanthodendrilla
sp.
Nat
Prod
Commun
2015;
10:
1945-‐1949
[2]
Sirimangkalakitti
N,
Yokoya
M,
Chamni
S,
Chanvorachote
P,
Plubrukrn
A,
Saito
N,
Suwanborirux
K.
Synthesis
and
absolute
configuration
of
acanthodendrilline,
a
new
cytotoxic
bromotyrosine
alkaloid
from
the
Thai
marine
sponge
Acanthodendrilla
sp.
Chem
Pharm
Bull
2016;
64:
258-‐262
[3]
Ingkaninan
K,
Temkitthawon
P,
Chuenchom
K,
Yuyaem
T,
Thongnoi
W.
Screening
for
acetylcholinesterase
inhibitory
activity
in
plants
used
in
Thai
traditional
rejuvenating
and
neurotonic
remedies.
J
Ethnopharmacol
2003;
89:
261-‐264
P588
Tools
and
strategies
to
access
to
original
bioactive
compounds
from
cultivation
of
marine
invertebrates
and
associated
symbi-‐
onts:
The
case
of
TASCMAR
Nikolas
Fokialakis1,
Ioannis
Trougakos1,
Yehuda
Benayahu2,
Suchana
Chavanich3,
Anne
Bi-‐
alecki4,
Michael
Schaeffer5,
Stefano
Zucchinali6,
Doru
Felezeu7,
Konstantinos
Gardikis8,
Ped-‐
ro
Álvarez9,
Åke
Lignell10,
Antonella
Passani11,
Asaf
Ariel12,
Jamal
Ouazzani13
1
University
of
Athens,
Athens,
Greece,
2
Tel
Aviv
University,
Tel
Aviv,
Israel,
3
Chulalongkorn
University,
Bangkok,
Thailand,
4
Université
de
la
Réunion,
Ile
de
la
Réunion,
France,
5
Crelux,
Martinsried,
Germany,
6
Bict,
Lodi,
Italy,
7
Pierre
Guerin
Technologies,
Mauzé-‐sur-‐le-‐Mignon,
France,
8
Apivita,
Athens,
Greece,
9
iMare
Natural,
Granada,
Spain,
10
AstaReal,
Gustavsberg,
Sweden,
11
T6
Ecosystems,
Rome,
Italy,
12
EcoOcean,
Kibbutz
Sdo
Yam,
Israel,
13
CSN-‐CNRS,
Gif
sur
Yvette,
France
TASCMAR
is
a
European
Commission
funded
project
under
H2020,
aspiring
to
develop
new
tools
and
strategies
to
overcome
existing
bottlenecks
in
the
discovery
and
industrial
exploita-‐
tion
of
marine-‐derived
biomolecules
(secondary
metabolites
and
enzymes)
with
applications
in
the
pharmaceutical,
nutraceutical
and
cosmeceutical
industries.
Exploitation
of
neglected
and
underutilised
marine
invertebrates
and
symbionts
from
the
mesophotic
zone
has
been
combined
with
innovative
approaches
for
the
cultivation
and
extraction
of
marine
organisms,
from
lab
to
pilot-‐scale,
including
the
construction
of
new
biotechnological
equipment.
This
approach
will
ensure
the
sustainable
supply
of
biomass
while
promoting
the
production
of
high
added
value
bioactive
marine
compounds.
State-‐of-‐the-‐art
analytical
instrumentation
and
in-‐house
databases
have
been
employed
for
metabolomics
analysis,
dereplication
and
characterisation
of
valuable
compounds.
In
addition
a
focused
panel
of
in-‐vitro,
cell-‐based,
in-‐
ovo
and
in-‐vivo
bioassays,
for
discovering
metabolites
with
anti-‐ageing
and/or
angiogenesis
modulating
activity,
are
guiding
the
project’s
workflow
to
reveal
the
lead
compounds.
Also
the
catalytic
potential
of
mesophotic
symbionts
and
deriving
enzyme
candidates
will
be
evaluated
in
the
fine
chemicals
and
bioremediation
industries.
TASCMAR
is
continuously
evaluated
for
its
socioeconomic
and
environmental
impact
in
order
to
balance
industrial
development
and
sustainable
growth.
The
project
also
aims
to
develop
higher
standards
for
bioprospecting
in
areas
of
rich
marine
biodiversity.
To
achieve
its
goals,
thirteen
partners
from
eight
countries
are
combining
their
efforts,
bringing
together
complementary
skills
and
giving
the
project
an
international
profile.
TASCMAR
involves
five
academic
institutions,
six
industrial
partners,
one
consulting
company,
one
NGO
and
has
a
total
budget
of
6.7
M€.
Acknowledgements:
This
work
has
been
financially
supported
by
EU
under
the
frame
of
TASCMAR
project
(H2020-‐BG-‐03-‐2014,
Grant
agreement: 634674).
Keywords: Mesophotic coral ecosystems, age-‐related diseases, marine invertebrate holobi-‐
onts, marine microbial symbionts, marine secondary metabolites.
P589
Marine
halogenated
compound
analysis:
from
an
R
package
to
the
isolation
of
new
griseophenone
derivatives
Catherine
Roullier1,
2,
Yann
Guitton1,
3,
Soizic
Prado4,
Olivier
Grovel1,2,
Yves
François
Pouchus1
1
Groupe
Mer,
Molécules,
Santé-‐EA
2160,
UFR
des
Sciences
Pharmaceutiques
et
Biologiques,
Université
de
Nantes,
9
Rue
bias,
44000
Nantes,
France,
2
Biogenouest,
Corsaire-‐Thalassomics,
Nantes,
France,
3
LU-‐
NAM
Université,
Oniris,
Laboratoire
d’Étude
des
Résidus
et
Contaminants
dans
les
Aliments
(LABERCA),
44307
Nantes,
France,
4
Molécules
de
Communication
et
Adaptation
des
Micro-‐organismes,
UMR
7245
CNRS/MNHN,
Muséum
National
d’Histoire
Naturelle,
75231
Paris
Cedex
05,
France
A
collection
of
culture
extracts
obtained
from
several
marine-‐derived
fungal
strains
collected
on
the
French
Atlantic
coast
was
investigated
by
HPLC-‐HRMS
in
order
to
prospect
for
halo-‐
genated
compounds
and
to
identify
potentially
original
ones.
To
achieve
a
fast,
automated
and
efficient
data
analysis,
a
bioinformatics
tool
named
MeHaloCoA
(Marine
Halogenated
Com-‐
pound
Analysis)
was
developed
and
included
into
R
as
a
package.
After
extraction
from
the
metabolic
fingerprints
of
all
the
peaks
and
their
associated
mass
spectra,
a
mathematical
filter
based
on
mass
isotopic
profiles
allowed
the
selective
detection
of
halogenated
molecules.
In-‐
tegrating
MeHaloCoA
into
a
dereplication
approach
allowed
the
identification
of
known
and
new
halogenated
compounds
in
a
challenging
amount
of
time.
Subsequent
targeted
purifica-‐
tion
led
to
the
isolation
of
several
halogenated
metabolites
including
two
new
natural
prod-‐
ucts
with
bioactive
potential.
Acknowledgements:
The
authors
acknowledge
the
Region
des
Pays
de
la
Loire
for
funding
of
this
study
through
the
CHIMIMAR
program
and
the
technical
platform
Corsaire-‐ThalassOMICS
of
Biogenouest
Keywords: Halogenated Natural Products, dereplication, Mass Spectrometry, automated
method
P590
Fighting
antibiotic
resistance:
Resensitizing
agents
from
marine
derived
fungi
Paul
Barac1,
Henrik
Harms1,
Ina
Engels2,
Stefan
Kehraus1,
Tanja
Schneider2,
Gabriele
M.
Kö-‐
nig1*
1University
of
Bonn,
Institute
of
Pharmaceutical
Biology,
Nußallee
6,
53115
Bonn,
Germany;
2University
of
Bonn,
Institute
for
Pharmaceutical
Microbiology,
Meckenheimer
Allee
168,
53115
Bonn,
Germany;
*email:
g.koenig@uni-‐bonn.de
Antibiotic
resistance
development
is
an
urgent
world
health
concern,
aggravated
by
a
lack
of
novel
antibiotic
discovery.
ß-‐lactams
still
represent
the
cornerstone
for
treating
bacterial
in-‐
fections,
but
they
are
rapidly
rendered
useless
due
to
antibiotic
resistance
[1].
Since
most
clin-‐
ical
antibiotics
are
natural
products,
natural
sources
might
offer
relevant
solutions
for
this
crisis
[2].
We
target
marine
fungi
for
finding
new
anti-‐infectious
compounds.
The
latter
are
often
associated
with
algae,
sponges,
etc.,
and
capable
of
producing
interesting
bioactive
com-‐
pounds
to
fight
competitors.
This
project
is
focused
on
a
special
class
of
such
compounds,
termed
resensitizing
agents.
When
applied
together
with
antibiotics,
resensitising
agents
show
a
synergic
effect
and
can
regain
the
sensitivity
of
the
bacteria
towards
the
antibiotic
[3].
Xanthocillin
monomethyl
ether
(XMME)
was
found
to
be
a
resensitizing
agent.
It
was
isolated
by
activity-‐guided
fractionation
from
the
fungus
Dichotomomyces
cejpii,
associated
with
a
tropical
sponge.
XMME
showed
intrinsic
in
vitro
activity
towards
Staphylococcus
aureus
MSSA
(MIC
value=
2.5
µg/ml)
and
MRSA
(MIC
value=
20.0
µg/ml)
strains.
When
used
in
combina-‐
tion
with
ertapenem
(4.0
μg/ml),
the
latter
being
inactive
towards
the
MRSA
strain,
extraor-‐
dinary
synergy
was
observed,
resulting
in
a
MIC
value
of
<0.001
µg/ml.
This
result
showed
that
XMME
is
a
remarkable
resensitizing
agent.
Further
studies
are
needed
to
elucidate
the
exact
mechanism
of
action
for
XMME,
but
preliminary
results
show
interference
in
cell
wall
biosynthesis.
This
study
illustrates
that
developing
assays
and
techniques
to
isolate
and
test
the
resensitizing
agent
activity
of
marine
fungal
metabolites
does
offer
new
perspectives
in
regaining
efficiency
for
some
of
our
most
important
antibiotics.
Acknowledgements:
This
research
is
funded
by
DZIF,
Deutsches
Zentrum
für
Infektionsforschung,
Germa-‐
ny
and
supported
by
the
BIGS
DrugS,
Bonn
International
Graduate
School
of
Drug
Sciences,
Germany.
Keywords: Fungal natural products, antibiotic synergistic effects, antibiotic resistance
References:
[1] Center
for
Disease
Control
and
Prevention.
Antibiotic
Resistance
Threats;
CDC:
Atlanta,
GA,
2013,
http://www.cdc.gov/drugresistance/pdf/ar-‐threats-‐2013-‐508.pdf
[2] Brown
ED,
Wright
GD.
Antibacterial
drug
discovery
in
the
resistance
era.
Nature
2016;
529:336-‐343
[3] Abreu
AC,
McBain
AJ,
Simões
M.
Plants
as
sources
of
new
antimicrobials
and
resistance-‐
modifying
agents.
Nat
Prod
Rep
2012;
29:
1007-‐1021
P591
Aceropterin
A,
a
gersolane
diterpene
from
the
Carribean
sea
plume
Pseudopterogorgia
acerosa
(Pallas)
(Gorgonacea)
Paul
Scesa,
Lyndon
West
Department
of
Chemistry
and
Biochemistry,
Florida
Atlantic
University,
777
Glades
Rd.,
33431
Boca
Ra-‐
ton,
Florida,
United
States
The
gorgonian
octocoral
Pseudopterogorgia
acerosa
(Pallas)
(Gorgonacea)
has
been
proven
a
rich
source
of
biologically
active
marine
natural
products
[1-‐4].
In
the
course
of
this
study
five
previously
reported
pseudopterane
and
cembrane
diterpenoids
(1-‐5)
have
been
isolated
[1-‐
4],
along
with
one
new
gersolane
diterpenoid,
aceropterin
A
(6).
This
is
the
first
reported
ger-‐
solane
to
be
isolated
from
P.
acerosa
and
is
similar
to
gersolide
(7)
isolated
from
Gersemia
rubiformis
(Ehrenberg)
(Nephtheidae)
[5].
Isolation
was
performed
using
reversed
phase
col-‐
umn
chromatography
and
RP-‐HPLC.
A
specimen
of
P.
acerosa
(55
g
dry
wt.)
collected
at
Sandy
Point,
Bahamas
was
extracted
with
methanol.
The
methanolic
extract
was
separated
into
five
fractions
on
polymeric
HP-‐20
resin
using
cyclic
loading
[6]
and
a
stepwise
gradient
of
Me2CO/H2O.
The
third
fraction
was
further
separated
into
twenty
fractions
by
column
chro-‐
matography
over
HP-‐20ss
using
Me2CO/H2O.
The
eighth
fraction
yielded
1,
while
the
tenth
yielded
3.
The
fifteenth
fraction
was
subjected
to
moderate
pressure
liquid
chromatography
over
C18
to
afford
mixtures
of
4
and
5
as
well
as
2
and
6.
Compounds
4
and
5
were
separated
by
preparative
HPLC
over
PRP-‐1,
while
the
mixture
of
2
and
6
was
purified
using
sequential
preparative
HPLC
over
PRP-‐1
and
semi-‐preparative
HPLC
over
C18
to
afford
2
and
6
(1
mg).
Structural elucidation was performed using HRESIMS along with 1D and 2D NMR, including
gCOSY, gHSQC, gHMBC and NOESY experiments. Herein we present the isolation and structural
elucidation of this rare diterpenoid. Further studies are in progress to characterize the biological
activity of this compound.
Acknowledgements:
This
work
supported
financially
by
grants
from
the
Florida
Atlantic
University
Office
of
Undergraduate
Research
and
the
American
Society
of
Pharmacognosy.
Keywords: Marine natural products, octocoral, Pseudopterogorgia acerosa, diterpenoid, cem-‐
brane, pseudopterane, gersolane
References:
[1] Bandurraga
MM,
Fenical,
W.
Pseudopterolide,
an
irregular
diterpenoid
with
unusual
cyto-‐
toxic
properties
from
the
Caribbean
sea
whip
Pseudopterogorgia
acerosa
(Pallas)
(Gorgo-‐
nacea).
J
Am
Chem
Soc
1982;
104:
6463−6465
[2] Look,
S.A.;
Burch,
M.T.;
Fenical,
W.;
Qi-‐tai,
Z.;
Clardy,
Jon.
Kallolide
A,
a
new
antiinflammato-‐
ry
diterpenoid,
and
related
lactones
from
the
Caribbean
octocoral
Pseudopterogorgia
kal-‐
los
(Bielschowsky).
J
Org
Chem
1985;
50:
5741−5746
[3] Wright
AE,
Burres
NS,
Schulte
GK.
Cytotoxic
cembranoids
from
the
gorgonian
Pseudopter-‐
ogorgia
bipinnata.
Tetrahedron
Lett
1989;
30:
3491–3494
[4] Roethle
PA,
Trauner
D.
The
chemistry
of
marine
furanocembranoids,
pseudopteranes,
ger-‐
solanes,
and
related
natural
products.
Nat
Prod
Rep
2008;
25:
298−317
[5] Williams
DE,
Andersen
RJ,
Parkanyi
L,
Clardy
J.
Gersolide,
a
diterpenoid
with
a
new
rear-‐
ranged
carbon
skeleton
from
the
soft
coral
Gersemia
rubiformis.
Tetrahedron
Lett
1987;
28:
5079−5080
[6] Houssen
WE,
Jaspars
M.
Methods
in
Biotechnology.
In:
Sarker
SD,
Latif
Z,
Gray
AI,
editors.
Natural
Products
Isolation,
Vol.
20.
New
Jersey:
Humana
Press;
2006:
353–391
P592
New
marine
sterols
from
a
Formosan
gorgonian
coral
Pinnigorgia
sp.
Yu-‐Chia
Chang1,2,
Chan-‐Shing
Lin1,3,
Jyh-‐Horng
Sheu1,3,
Ping-‐Jyun
Sung1,2,4
1
Doctoral
Degree
Program
in
Marine
Biotechnology,
National
Sun
Yat-‐sen
University
and
Academia
Sini-‐
ca,
Kaohsiung
804,
2
National
Museum
of
Marine
Biology
&
Aquarium,
Pingtung
944,
Taiwan,
3
Depart-‐
ment
of
Marine
Biotechnology
and
Resources,
National
Sun
Yat-‐sen
University,
Kaohsiung
804,
4
Gradu-‐
ate
Institute
of
Marine
Biology,
National
Dong
Hwa
University,
Pingtung
944,
Taiwan
A
large
number
of
steroids
have
been
isolated
from
marine
organisms
and
many
of
them
have
been
shown
to
be
bioactive
[1].
In
continuation
of
our
studies
[2,3],
four
new
marine
sterols,
(22E,24R)-‐ergosta-‐5,22-‐diene-‐3β,11α-‐diol
(1),
(24S)-‐ergosta-‐5-‐ene-‐3β,11α-‐diol
(2),
5α,6α-‐
epoxy-‐23-‐demethylgorgost-‐8-‐ene-‐3β,7α-‐diol
(3)
and
5α,6α-‐epoxy-‐23-‐demethylgorgost-‐8(14)
-‐ene-‐3β,7α-‐diol
(4),
along
with
a
known
metabolite,
23-‐demethylgorgost-‐7-‐ene-‐3β,5α,6β-‐triol
(5),
were
isolated
from
an
algal-‐bearing
gorgonian
coral
Pinnigorgia
sp.,
collected
off
the
wa-‐
ters
of
Taiwan.
The
structures
were
elucidated
on
the
basis
of
their
spectroscopic
data.
The
sterols
1–5
were
tested
for
in
vitro
cytotoxicity
in
hepatic
stellate
cells
(HSCs).
Proliferation
of
HSCs
plays
a
key
role
in
the
pathogenesis
of
liver
fibrosis.
Figure
1.
The
structures
of
sterols
1–5.
Keywords:
Sterol,
Gorgonian,
Pinnigorgia,
Hepatic
Stellate
Cells,
Zooxanthellae
Reference:
[1] Blunt
JW,
Copp
BR,
Keyzers
RA,
Munro
MHG,
Prinsep
MR.
Marine
natural
products.
Nat
Prod
Rep
2016:
33:
382-‐431
[2] Chang
YC,
Kuo
LM,
Hwang
TL,
Yeh
J,
Wen
ZH,
Fang
LS,
Wu
YC,
Lin
CS,
Sheu
JH,
Sung
PJ.
Pin-‐
nisterols
A–C,
new
9,11-‐secosterols
from
a
gorgonian
Pinnigorgia
sp.
Mar
Drugs
2016:
14:
12
[3] Chang
YC,
Kuo
LM,
Su
JH,
Hwang
TL,
Kuo
YH,
Lin
CS,
Wu
YC,
Sheu
JH,
Sung
PJ.
Pinnigorgiols
A–C,
9,
11-‐secosterols
with
a
rare
arrangement
from
a
gorgonian
coral
Pinnigorgia
sp.
Tetrahedron
2016:
72:
999-‐1004
P593
Anti-‐microbial
metabolites
from
a
marine
bacterium
YMA4
Pi-‐Yu
Chen,
Ning
Lu,
Ying-‐Mi
Lai,
Yu-‐Lang
Yang*
Agricultural Biotechnology Research Center, Academia Sinica, 11529 Taipei, Taiwan
Among
studies
exploring
new
anti-‐microbial
agents
from
marine
microbial
sources,
a
Gram-‐
negative
bacterium
YMA4
was
isolated.
In
the
agar-‐plug
diffusion
assay,
YMA4
showed
a
strong
inhibition
effect
against
Staphylococcus
epidermidis,
Paenibacillus
larvae,
and
Candida
albicans,
together
with
a
middle
inhibition
effect
against
Staphylococcus
aureus.
Using
MALDI
IMS
[1]
and
LC
MS/MS
molecular
network
analysis
[2],
we
identified
the
major
anti-‐bacterial
metabolites
of
YMA4
as
cyclic-‐lipodepsipeptide
empedopeptin
[3,4]
together
with
several
new
analogues.
However,
unlike
the
consistent
and
significant
inhibition
effect
shown
in
the
antagonistic
assay,
the
inhibition
potency
of
YMA4
extract
against
C.
albicans
was
somehow
unstable,
which
implies
the
interaction
between
YMA4
and
C.
albicans
is
too
complicated
to
be
explored
through
the
classic
bioactivity
guided
fractionation
and
isolation
approach.
Through
the
genome
mining
and
HPLC
PDA
profiling
analysis,
we
successfully
isolated
and
identified
the
unstable
anti-‐fungal
metabolites
of
YMA4
as
polyyne
analogues.
In
conclusion,
the
Gram-‐
negative
bacterium
YMA4
has
a
potent
anti-‐microbial
effect
against
not
only
Gram-‐positive
pathogens,
S.
aureus,
S.
epidermidis,
and
P.
larvae
but
also
pathogenic
fungus,
C.
albicans,
by
utilizing
different
types
of
metabolites.
Acknowledgements:
This
work
was
supported
by
grants
from
the
Ministry
of
Science
and
Technology,
Taiwan,
to
Y.-‐L.Y.
(MOST
104-‐2320-‐B-‐001-‐019-‐MY2).
Keywords: cyclic-‐lipodepsipeptide, polyyne, MALDI IMS, molecular networking, genome min-‐
ing
References:
[1] Shih
CJ,
Chen
PY,
Liaw
CC,
Lai
YM,
Yang
YL.
Bringing
microbial
interactions
to
light
using
imaging
mass
spectrometry.
Nat
Prod
Rep
2014;
31:
739-‐755.
[2] Yang
JY,
Sanchez
LM,
Rath
CM,
Liu
X,
Boudreau
PD,
Bruns
N,
Glukhov
E,
Wodtke
A,
de
Fe-‐
licio
R,
Fenner
A,
Wong
WR,
Linington
RG,
Zhang
L,
Debonsi
HM,
Gerwick
WH,
Dorrestein
PC.
Molecular
networking
as
a
dereplication
strategy.
J
Nat
Prod
2013;
76:
1686-‐1699.
[3] Konishi
M,
Sugawara
K,
Hanada
M,
Tomita
K,
Tomatsu
K,
Miyaki
T,
Kawaguchi
H,
Buck
RE,
More
C,
Rossomano
VZ.
Empedopeptin
(BMY-‐28117),
a
new
depsipeptide
antibiotic.
I.
Production,
isolation
and
properties.
J
Antibiot
1984;
37:
949-‐957.
[4] Sugawara
K,
Numata
K,
Konishi
M,
Kawaguchi
H.
Empedopeptin
(BMY-‐28117),
a
new
depsipeptide
antibiotic.
II.
Structure
determination.
J
Antibiot
1984;
37:
958-‐964.
P594
Bioactivity
guided
isolation
of
secondary
metabolites
from
a
red
green
Hawaiian
sponge
Ram
P.
Neupane1,
Stephen
Parrish1,
Wesley
Yoshida1,
John
Head1,
Richard
Yip2,
James
Turk-‐
son2,
Philip
Williams1,2
1Department
of
Chemistry,
University
of
Hawaii
at
Manoa,
Honolulu,
HI
96822,
USA,
2University
of
Hawaii
Cancer Center, Honolulu, HI 96813, USA
Marine sponges are rich sources of bioactive secondary metabolites. In our quest for the dis-‐
covery of the inhibitors of BACE1 (b-‐site of Amyloid Precursor Protein Cleaving Enzyme), an
enzyme implicated in the pathogenesis of Alzheimer’s disease [1,2], we investigated a current-‐
ly unidentified red green sponge collected from the Sheraton Caverns in Kauai. Bioactivity
guided isolation of this sponge (11-‐Kauai-‐80) led to the discovery of several known sesquit-‐
erpenoid quinones (1-‐6) and hydroquinones (7-‐9), and a new 11-‐membered cyclic compound
(10) with a unique structural framework. The relative configuration of 10 was established
from density functional theory (DFT) calculations of GIAO NMR shielding tensors for all four
possible diastereomers of a truncated molecule, where the alkyl side chains were replaced by
methyl groups, and statistical comparison of the experimental and computed 1H and 13C
chemical shifts. The absolute configuration of the cyclic molecule was established from the
advanced Marfey’s analysis of an NMeLeu residue following hydrolysis of the intact molecule.
A
few
of
these
known
metabolites
(1,
3
and
7)
showed
moderate
activity
against
BACE1.
For
example,
ilimaquinone
(1)
inhibited
BACE1
with
an
IC50
value
of
65
μM.
Additionally,
com-‐
pounds
1-‐9
exhibited
moderate
to
strong
inhibition
of
viability
of
U251
(human
glioma)
cells.
Smenospongine
(3)
and
dictyoceratin
A
(7)
showed
the
most
potent
activity,
with
IC50
values
of
2.4
μM
and
2.8
μM,
respectively.
Keywords: BACE1, human glioma, sesquiterpenoid quinones, macrolide
References:
[1] Hardy
J,
Higgins
G.
Alzheimer’s
disease:
the
amyloid
cascade
hypothesis.
Science
1992;
256:
184−185
[2] Goedert
M,
Spillantini
MG.
A
century
of
Alzheimer’s
disease.
Science
2006;
314:
777−781
P595
Effect
of
alkaloids
isolated
from
Haliclona
sp.
against
hydrogen
peroxide-‐induced
injury
in
SH-‐SY5Y
human
neuroblastoma
cells
Rebeca
Alvariño1,
Eva
Alonso1,
Marie-‐Aude
Tribalat2,
Olivier
P.
Thomas2,
3,
Luis
M.
Botana1
1
Departamento
de
Farmacología,
Facultad
de
Veterinaria,
Universidad
de
Santiago
de
Compostela,
Lugo
27003,
Spain,
2
Géoazur
UMR
Université
Nice
Sophia
Antipolis-‐CNRS-‐IRD-‐OCA,
250
Avenue
Albert
Einstein
06560
Valbonne,
France,
3
National
University
of
Ireland
Galway,
Marine
Biodiscovery,
School
of
Chemis-‐
try,
University
Road,
Galway,
Ireland
The
great
biodiversity
of
the
oceans
makes
the
marine
environment
a
rich
source
of
new
bio-‐
active
compounds.
Particularly,
marine
sponges
have
provided
several
secondary
metabolites
with
potential
pharmaceutical
applications.
Sarains
are
diamide
alkaloids
isolated
from
the
Mediterranean
sponge
Haliclona
(Rhizoniera)
sarai
that
have
already
showed
antibacterial,
insecticidal
and
anti-‐fouling
activity.
In
this
study,
we
examined
for
first
time
the
neuroprotec-‐
tive
effects
of
sarains
1,
2
and
A
against
oxidative
stress.
With
this
purpose,
sarains
were
test-‐
ed
in
an
in
vitro
oxidative
stress
model
using
human
neuroblastoma
SH-‐SY5Y
cells.
Com-‐
pounds
were
co-‐incubated
with
hydrogen
peroxide
for
6
hours
and
protective
effects
were
evaluated.
Sarain
A
was
the
most
promising
compound,
improving
mitochondrial
function
and
decreasing
reactive
oxygen
species
levels
(ROS).
In
view
of
these
results,
the
ability
of
sarain
A
to
induce
the
nuclear
factor
E2-‐related
factor
2
(Nrf2)-‐antioxidant
response
element
pathway
was
determined.
This
compound
enhanced
Nrf2
translocation
to
the
nucleus,
which
suggests
that
sarain
A
is
acting
as
an
indirect
antioxidant.
Oxidative
stress
produces
mitochondrial
dysfunction,
which
is
related
to
neurodegenerative
disorders
as
Alzheimer’s,
Parkinson’s
and
Huntington
diseases.
Therefore,
diminishing
ROS
release
and
improving
antioxidant
systems
might
be
a
potential
therapeutic
strategy
against
these
illnesses.
Indirect
antioxidants,
more
than
direct
ones,
are
considered
a
promising
tool
to
decrease
oxidative
stress
because
they
can
induce
the
expression
of
cytoprotective
proteins
and
reduce
mitochondrial
dysfunction.
Our
results
indicate
that
sarain
A
may
be
a
candidate
compound
for
further
studies
in
neurodegenerative
diseases.
Keywords:
Oxidative
stress,
sarains,
neurodegenerative
diseases,
Nrf2
P596
Latrunculid
sponges,
their
microbial
communities
and
secondary
metabolites:
connecting
conserved
bacterial
symbionts
to
pyr-‐
roloiminoquinone
production.
Rosemary
A
Dorrington1,
Storm
H
Hilliar1,
Jarmo
CJ
Kalinski1,
Rui
WM
Krause2,
Kerry
L
McPhail3,
Shirley
Parker-‐Nance1,
Tara
A
Wlamsley4,
Samantha
C
Waterworth1
1
Department
of
Biochemistry
and
Microbiology,
Rhodes
University,
Grahamstown
South
Africa.
2
De-‐
partment
of
Chemistry,
Rhodes
University,
Grahamstown
South
Africa.
3
School
of
Pharmacology,
Oregon
State
University,
Corvallis,
USA.
4
Department
of
Biotechnology,
Vaal
University
of
Technology,
Van-‐
derbijlpark,
South
Africa
The
Latrunculiidae
are
cold
water
sponges
known
for
their
production
of
bioactive
pyr-‐
roloiminoquinone
alkaloids
(e.g.
makaluvamines,
discorhabdins
and
tsitsikammamines).
Since
pyrroloiminoquinones
have
also
been
isolated
from
sponges
belonging
to
other
families,
ascidians
and
microorganisms,
the
biosynthetic
origin
of
these
alkaloids
in
latrunculid
spong-‐
es
is
likely
microbial.
This
study
focuses
on
the
secondary
metabolites
produced
by
closely-‐
related
Tsitsikamma
species
and
Cyclacanthia
bellae,
all
latrunculid
sponges
endemic
to
Algoa
Bay
on
the
South
African
southeast
coast.
The
sponges
produced
suites
of
related
pyr-‐
roloiminoquinones,
including
tsitsikammine
A
and
B,
and
discohabdin
C
and
V,
the
combina-‐
tion
and
relative
abundance
of
which
is
species-‐specific.
Characterisation
of
the
diversity
of
sponge-‐associated
bacterial
communities
revealed
the
unprecedented
conservation
of
two
dominant
bacterial
species.
The
first,
a
Betaproteobacterium,
is
also
found
in
other
latruncu-‐
lids
and
related
sponge
families,
representing
a
novel
clade
of
sponge
endosymbionts
that
have
co-‐evolved
with
their
hosts.
The
second
conserved
bacterial
symbiont
is
a
spirochaete
found
only
in
Cyclacanthia
and
Tsitsikamma
species
that
is
likely
to
have
been
recruited
from
free-‐living
spirochaetes
in
the
environment.
This
study
sheds
new
light
on
the
interactions
between
latrunculid
sponges,
their
dominant
bacterial
symbionts,
and
the
potential
involve-‐
ment
of
these
bacteria
in
pyrroloiminoquinone
biosynthesis.
Keywords: Pyrroloiminoquinones, bacterial phylogenies, conserved sponge symbionts
P597
Defensive
chemistry
of
the
Irish
nudibranch
Archidoris
psuedoargus
(Gastropoda
opisthobranchia)
Ryan
M.
Young1,2,
Bill
J.
Baker1,2
1
School
of
Chemistry,
National
University
of
Ireland
Galway,
Galway,
Republic
of
Ireland;
2
Department
of
Chemistry
and
Center
for
Drug
Discovery
and
Innovation,
University
of
South
Florida,
Tampa,
33620,
FL,
USA.
Historically,
marine
natural
products
from
the
Republic
of
Ireland
have
been
greatly
un-‐
derrepresented
in
the
literature
despite
having
a
coastline
of
over
4500
miles.
Archidoris
pseudoargus
is
a
soft-‐bodied,
slow
moving
Dorid
nudibranch
which
inhabits
the
coastal
wa-‐
ters
of
Ireland
and
the
United
Kingdom.
Nudibranchs
are
a
good
source
of
new
chemical
di-‐
versity,
employing
these
secondary
metabolites
to
deter
predation.
In
this
study
we
have
identified
new
chemistry
as
well
as
used
a
metabolomics
approach
to
identify
the
origin
of
said
chemistry
as
well
as
trends
in
morphologically
different
individuals.
From
our
initial
find-‐
ings
the
individual
organisms
group
together
into
several
clusters
based
on
their
metabolom-‐
ic
profiles
(Figure
1
right).
The
major
metabolites
isolated
thus
far
from
these
organisms
are
diterpene
glycerides
(e.g.,
1
and
2)
which
have
been
shown
to
be
icthyotoxins
[1]
suggesting
these
metabolites
are
the
nudibranchs
method
of
defense
against
predation.
In
early
Spring,
mature
adults
come
together
to
reproduce
and
shortly
thereafter
to
oviposit
on
the
subtidal
rocky
shoreline.
These
egg
sacs
can
be
brightly
coloured
and
are
readily
ex-‐
posed
to
predation,
yet
none
of
the
many
surrounding
predators
appear
to
feed
on
these
nu-‐
trient
rich
egg
masses.
Using
UPLCMS-‐MS
on
standards
isolated
from
the
bulk
A.
psuedoargus
extraction,
we
have
shown
that
the
same
diterpenes
present
in
the
mantle
tissue
of
the
adults
is
transferred
into
the
egg
masses
from
the
parent
organism
which
in
turn
protects
them
against
predators.
Figure
1:
Left:
Representative
A.
psuedoargus
chemistry.
Right:
MDS
plot
showning
differences
in
metabolomic
profiles
of
the
diterpenoids
(30
<
C
>
20)
of
individials
collected
from
the
same
location.
Solid
(>
50%)
and
broken
lines
(>
60%)
indicate
varing
degrees
of
similarity
between
data
points.
Acknowledgements:
Beaufort
Marine
Research
Award:
This
Beaufort
Marine
Research
Award
is
carried
out
under
the
Sea
Change
Strategy
and
the
Strategy
for
Science
Technology
and
Innovation
(2006-‐2013),
with
the
support
of
the
Marine
Institute,
funded
under
the
Marine
Research
Sub-‐Programme
of
the
National
Development
Plan
2007–2013.
Keywords: nudibranch, diterpenes, metabolomics, defensive chemistry
References:
[1] Cimino G, Gavagnin M, Sodano G, Puliti R, Mattia CA, Mazzarella L. Verrucosin-‐a and -‐b,
ichthyotoxic diterpenoic acid glycerides with a new carbon skeleton from the dorid
nudibranch Doris verrucosa. Tetrahedron 1988; 44, 2301-‐2310
P598
Serratia
marcescens
metabolites
are
promising
candidates
for
bi-‐
ocontrol
of
avocado
pathogens
S.
David
Granada1,
Juan
C.
Bedoya-‐Pérez
2,
Kirstin
Scherlach3,
Christian
Hertweck3
1Corporation
for
Biological
Research,
Medellín,
Colombia.
2Universitary
Institution
Mayor
College
of
An-‐
tioquia,
Medellín,
Colombia.
3Leibniz
Institute
for
Natural
Product
Research
and
Infection
Biology
(HKI),
Jena,
Germany.
Colletotrichum
spp.
and
Phytophthora
cinnamomi
can
cause
severe
economic
losses
in
avoca-‐
do
production
in
postharvest
as
well
as
in
crop
stages.
Therefore,
world
avocado
agroindustry
is
demanding
for
new
alternatives
to
cope
with
these
pathogens.
The
main
objective
of
this
study
was
to
assess
the
production
and
inhibitory
activity
of
secondary
metabolites
from
the
well-‐known
biocontrol
bacterium
Serratia
marcescens
against
avocado
pathogens.
S.
mar-‐
cescens
strain
ARP5.1
was
obtained
from
the
rhizoplane
of
an
apparently
healthy
avocado
tree.
A
bioactivity-‐guided
methodology
was
used
to
isolate
molecules
of
interest.
Minimal
me-‐
dium
standardization
was
performed
in
order
to
increase
the
overall
activity
of
the
crude
ex-‐
tract.
One
factor
at
a
time
approach
and
a
Plackett-‐Burman
(PB)
design
were
carried
out
for
the
selection
of
appropriate
carbon
and
nitrogen
sources
and
the
most
influencing
fermenta-‐
tion
parameter
among
nine
variables.
As
a
result,
three
active
compounds
(prodigiosin,
ser-‐
ratamolide
and
haterumalide
NA)
were
isolated
from
the
crude
extract
and
structurally
eluci-‐
dated.
Haterumalide
NA
proved
to
be
the
most
active
metabolite
with
a
MIC
of
5
and
<0.16
µg
mL-‐1
against
Colletotrichum
gloeosporioides
and
P.
cinnamomi,
respectively.
Prodigiosin
(6.25
and
6.25
µg
mL-‐1)
and
serratamolide
(80
and
80
µg
mL-‐1)
were
less
active.
Highest
production
of
haterumalide
NA
was
achieved
in
culture
media
consisting
of
maltose
and
yeast
extract
and
PB
experiments
revealed
maltose
concentration,
pH
and
Mg2+
as
the
most
influencing
factors
for
haterumalide
NA
production
(p<0.05).
With
these
optimisations
the
production
rate
could
be
increased
16.7
fold
compared
with
the
basic
minimal
medium
composition.
Our
results
show
that
haterumalide
NA
is
a
highly
effective
compound
for
the
control
of
avocado
diseases
and
a
promising
candidate
to
become
a
commercially
available
product
due
to
its
easy
pro-‐
duction
and
strong
antioomycete
activity.
Acknowledgements:
Colciencias
and
General
Royalties
System
from
Colombia.
Keywords: Haterumalide, Phytophthora cinnamomi, antioomycete, Plackett-‐Burman, Colleto-‐
trichum gloeosporioides.
References:
[1] Fu
XT,
Lin
H,
Kim
SM.
Optimization
of
medium
composition
and
culture
conditions
for
agarase
production
by
Agarivorans
albus
YKW-‐34.
Process
Biochem
2009;
44:
1158–
1163.
[2] Massart
S,
Martinez-‐Medina
M,
Haissam
JM.
Biological
control
in
the
microbiome
era:
chal-‐
lenges
and
opportunities.
Biol
Control
2015;
89:
98–108.
[3] Ramírez
S,
Arias
JD,
Bedoya
JC,
Rueda
EA,
Sánchez
CY,
Granada
SD.
Metabolites
produced
by
antagonistic
microbes
inhibit
the
principal
avocado
pathogens
in
vitro.
Agron
Colomb
2015;
3:
58–63.
P599
Potential
Antimicrobial
Activities
of
Seagrasses,
Zostera
spp.
from
Aegean
Sea;
West
Coast
of
Turkey
Gülçin
Saltan
İşcan1,
Özlem
Bahadır
Acıkara1,
Müjde
Eryılmaz2,
Burçin
Ergene1,
Serkan
Özbilg-‐
in1,
Sertel
Seçer3,
Hijran
Yavuzcan3
1
Department
of
Pharmacognosy,
Ankara
University,
Faculty
of
Pharmacy,
06100
and
Ankara,
Turkey,
2
Department
of
Pharmaceutical
Microbiology,
Ankara
University,
Faculty
of
Pharma-‐
cy,
06100
and
Ankara,
Turkey,
3
Department
of
Fisheries
and
Aquaculture,
Ankara
University,
Faculty
of
Agriculture,
06110
and
Ankara,
Turkey
Marine
and
estuarine
submersed
aquatic
angiosperms
or
seagrasses
produce
antimicrobial
compounds
that
may
act
to
reduce
or
control
microbial
growth
[1].
Zostera
spp.
being
one
of
the
most
widely
distributed
plant
in
marine
environment
is
an
important
species
in
coastal
ecosystems
with
contributing
to
nutrient
cycling
and
sediment
stabilizer,
providing
food
stuffs
and
habitat
for
many
marine
organisms
[2].
The
aim
of
this
study
is
to
evaluate
the
an-‐
timicrobial
activity
of
extracts
from
Zostera
spp.
collected
from
Aegean
SeaCoasts.
Dead
leaves
of
Zostera
marina
and
Z.
noltii
were
collected
from
Urla
district
near
Izmir
bay
(38°19’47.9”
N
and
26°39’10.9”
E)
in
autumn.
Taxononomic
identification
of
the
plant
was
confirmed
by
refe-‐
rence
to
literature.
Extracts
were
prepared
by
maceration
at
room
temperature
succesi-‐
vely
using
n-‐hexane,
ethylacetate
and
methanol.
All
solvents
were
evaporated
under
vacuum
to
obtain
crude
dry
residues.
DMSO
in
water
(20%)
was
used
to
dissolve
the
extracts
before
testing
for
their
antimicrobial
activities.
In
this
study,
methanol,
n-‐hexane
and
ethylacetate
extracts
of
Zostera
spp.
which
were
prepared
by
maceration
at
room
temperatu-‐
re,
were
screened
for
their
potential
in
vitro
antibacterial
activities
against
Staphylococcus
aureus
ATCC
29213,
Enterococcus
faecalis
ATCC
29212,
Bacillus
subtilis
ATCC
6633,
Escheric-‐
hia
coli
ATCC
25922,
Pseudomonas
aeruginosa
ATCC
27853
and
antifungal
activities
against
Candida
albicans
ATCC
10231.
Micro
broth
dilution
method
was
used
for
determination
of
the
minimum
inhibitory
concentrations
(MIC).
Serial
two-‐fold
dilutions
ranging
from
0.078-‐10
mg/mL
were
prepared
in
medium.
A
set
of
wells
containing
only
inoculated
broth
was
used
as
control.
After
incubation
for
18-‐24
h
at
37±1
0C
for
bacteria-‐48
h
for
fungi,
the
last
well
with
no
microbial
growth
was
recorded
to
represent
MIC
value
(mg/mL)
[3,4].
Ethyl
acetate
extract
of
Zostera
spp.
exhibited
better
antimicrobial
activity
than
n-‐hexane
and
methanol
extracts
(5
mg/mL).
No
antibacterial
activity
was
observed
against
B.
subtilis
and
E.
coli
for
all
extracts.
The
present
study
contributes
to
the
identification
of
seagrasses
from
the
Aeagean
Sea
coasts
that
exhibit
antimicrobial
properties.
Acknowledgements:
This
study
was
supported
by
TUBITAK
(The
Scientific
and
Technical
Research
Coun-‐
cil
of
Turkey
(Project
Number:
114Y141).
Keywords: Seagrasses, Zostera spp., antimicrobial activity.
References:
[1] Kumar CS, Sarada DVL, Gideon TP, Rengasamy R. Antibacterial activity of three South Indi-‐
an seagrasses, Cymodocea serrulata, Halophila ovalis and Zostera capensis. World J Microb
Biot 2008; 24: 1989-‐1992
[2] Boschker HTS, Wielemaker A, Schaub BEM, Holmer M. Limited coupling of macrophyte
production and bacterial carbon cycling in the sediments of Zostera spp. meadows. Mar
Ecol Prog Ser 2000; 203: 181-‐189
[3] Wayne
PA.
Methods
for
the
dilution
antimicrobial
susceptibility
tests
for
bacteria
that
grow
aerobically,
Approved
standard,
8th
edition.
USA:
Clinical
and
Laboratory
Standards
Institute
M7-‐A8
Publication;
2009
[4] European
Committee
on
Antimicrobial
Susceptibility
Testing
(EUCAST).
Breakpoint
tables
for
interpretation
of
MICs
and
zone
diameters.
Version
3.1
P600
New
diterpene
isolated
from
a
sponge
of
genus
Strongylophora
Stephen
M.
Parrish,
Wesley
Y.
Yoshida,
Philip
G.
Williams
Department of Chemistry, University of Hawaii at Mānoa, Honolulu, Hawaii, 96822, U.S.A.
In
our
quest
for
discovering
new
and
novel
marine
natural
products,
two
diterpenes
(1-‐2)
were
isolated
from
a
Hawaiian
sponge
of
the
genus
Strongylophora,
which
was
collected
off
the
southern
coast
of
Lana'i.
New
diterpene
(1)
contains
an
uncommon
naphthyl
A,
B
ring
system
similar
to
equilenin
[1],
one
of
the
active
components
of
the
FDA
approved
drug
Premarin,
but
also
unusual
oxidation
at
position
2.
The
planar
structure
of
(1)
was
deduced
by
analysis
of
2D
NMR
spectra
(COSY,
HMBC,
HSQC,
ROESY)
and
high
resolution
mass
spec-‐
trometry
data.
CD
spectra
of
(1)
displayed
a
negative
cotton
effect
in
accordance
with
predic-‐
tions
made
using
the
octant
rules
[2],
suggesting
an
S
configuration
of
the
lone
stereocenter.
Cinanthrenol
A
(2)
was
previously
isolated
from
a
sponge
of
genus
Cinachyrella
and
demon-‐
strated
bioactivity
against
P-‐388,
HeLa
cells,
and
as
an
estrogen
receptor
inhibitor
[3].
Struc-‐
tural
similarities
between
(1)
and
(2)
suggest
(1)
may
have
similar
biological
activity.
Acknowledgements:
Dr.
Shugeng
Cao
is
acknowledged
for
obtaining
CD
spectra
Keywords: Sponge metabolite, diterpene, CD
References:
[1] Girard
A,
Sandulesco
G,
Fridenson
A,
Gaundefroy
C,
Rutgers
JJ.
A
New
Crystalline
Sex
Hor-‐
mone.
Compt
Rend
1932;
195:
981-‐983
[2] Moffitt
W,
Woodward
RB,
Moscowitz
A,
Klyne
W,
Djerassi
C.
Structure
and
the
Optical
Rota-‐
tory
Dispersion
of
Saturated
Ketones.
J
Am
Chem
Soc
1961;
83:
4013-‐4018
[3] Machida
K,
Abe
T,
Arai
D,
Okamoto
M,
Shimizu
I,
de
Voogd
NJ,
Fusetani
N,
Nakao
Y.
Cinan-‐
threnol
A,
an
Estrogenic
Steroid
Containing
Phenanthrene
Nucleus,
from
a
Marine
Sponge
Cinachyrella
sp.
Org
Lett
2014;
16:
1539-‐1541
P601
New
polyketides
from
dinoflagellate
Amphidinium
sp.
Takaaki Kubota1,2, Takahiro Iwai2, Hayato Sato2, Jun’ichi Kobayashi2
1 Showa Pharmaceutical University, 3-‐3165 Higashi-‐Tamagawagakuen, Machida, Tokyo, 194-‐8543, Ja-‐
pan, 2 Graduate School of Pharmaceutical Sciences, Hokkaido University, Sapporo 060-‐0812, Japan
Marine
dinoflagellates
have
been
recognized
as
a
source
of
novel
secondary
metabolites
with
intriguing
structures
and
interesting
bioactivities
[1].
In
our
continuing
search
for
bioactive
metabolites
from
Okinawan
marine
dinoflagellates,
we
have
isolated
a
series
of
macrolides,
amphidinolides,
and
long
chain
polyketides
from
cultured
symbiotic
marine
dinoflagellates
Amphidinium
spp.
[2].
Recently,
we
have
investigated
the
culture
medium
of
dinoflagellate
Amphidinium
sp.,
which
was
a
symbiont
of
an
acoel
flatworm
Amphiscolops
sp.,
and
isolated
five
new
liner
polyketides,
amphidinins
C~G
(1~5),
with
known
polyketides,
amphidinin
A
(6)
and
amphidinolide
Q
(7)
[3-‐5].
The
analysis
of
spectral
data
revealed
that
1~4
were
4,5-‐
seco-‐analogs
of
7.
The
absolute
configurations
of
1~4
have
been
elucidated
by
J-‐based
config-‐
uration
analysis
(JBCA),
modified
Mosher’s
method,
and
chemical
derivatization.
Compounds
1~4
corresponded
to
4,5-‐secoamphidinolide
Q,
4-‐O-‐α-‐D-‐ribofuranosyl-‐4,5-‐secoamphidinolide
Q,
6,O-‐6-‐dihydro-‐4,5-‐secoamphidinolide
Q,
and
4-‐O-‐α-‐D-‐ribo-‐furanosyl-‐6,O-‐6-‐dihydro-‐4,5-‐
secoamphidinolide
Q,
respectively.
Compounds
2
and
4
were
the
first
glycoside
related
to
amphidinolides
[6].
The
structure
of
5
was
elucidated
to
be
9,12-‐O-‐seco-‐analog
of
6
based
on
its
spectral
data.
The
absolute
configuration
of
5
has
been
elucidated
to
be
2R,
4R,
6S,
11R,
and
12S
by
chemical
shift
comparison,
JBCA,
and
modified
Mosher’s
method
[7].
The
absolute
con-‐
figuration
of
6
was
also
elucidated
to
be
2R,
4R,
6S,
9R,
11R,
and
12S
by
JBCA,
modified
Mosher’s
method,
and
the
density-‐functional
theory
calculations
[8].
Compounds
1,
3,
and
7
showed
antimicrobial
activities
against
some
bacteria
and
fungi.
Compounds
2
(IC50
16
µg/mL),
4
(IC50
32
µg/mL),
and
5
(IC50
8
µg/mL)
showed
selective
antifungal
activity
against
Trichophyton
mentagrophytes.
Compound
6
showed
antibacterial
activity
against
Bacillus
sub-‐
tilis
(MIC
16
µg/mL)
and
antifungal
activity
against
Aspergillis
niger
(IC50
4
µg/mL).
18 19 18 19 18 19
21 21 21
7R 9S 7R 9S 7R 9S
6 16 6 S 16 6
11 13 11 13 11 13 16
5 O R R 5 OH R R 5 O R R
4 4 4 R
O 20
O 20 O 20
RO 1 RO 1 HO 1
17 O 17
O 17
O
1:R =H 3: R =H 7
2 : R = α-D-ribofuranosyl 4 : R = α-D-ribofuranosyl
19
21 20 19 21 18
11 R
R 13 17 R 9 12 S
R R S 7 9 11 S 16 R R S
HO 1
2 4 6 12 HO 1
2 4 6
7 O 13
16 17
20 H
OH OH 22 OH 18 OH OH 22
5 6
Acknowledgements:
Prof.
Tohru
Gonoi
and
Dr.
Kanae
Sakai
are
acknowledged
for
antimicrobial
activity
tests
Keywords: marine dinoflagellate, Amphidinium sp., polyketide
References:
[1] Blunt
JW,
Copp
BR,
Keyzers
RA,
Munro
MHG,
Prinsep
MR.
Marine
natural
products. Nat
Prod
Rep
2016,
33:
382-‐431
[2] Kobayashi
J.
Amphidinolides
and
its
related
macrolides
from
marine
dinoflagellates.
J
An-‐
tibiot
2008,
61:
271-‐284
[3] Kobayashi
J,
Yamaguchi
N,
Ishibashi
M.
Amphidinin
A,
a
novel
amphidinolide-‐related
me-‐
tabolite
from
the
cultured
marine
dinoflagellate
Amphidinium
sp.
Tetrahedron
Lett
1994,
35:
7049-‐7050
[4] Kobayashi
J,
Takahashi
M,
Ishibashi
M.
Amphidinolide
Q,
a
novel
12-‐membered
macrolide
from
the
cultured
marine
dinoflagellate
Amphidinium
sp.
Tetrahedron
Lett
1996,
37:
1449-‐1450
[5] Takahashi
Y,
Kubota
T,
Fukushi
E,
Kawabata
J,
Kobayashi
J.
Absolute
stereochemistry
of
amphidinolide
Q.
Org
Lett
2008,
10:
3709-‐3711
[6] Kubota
T,
Iwai
T,
Sakai
K,
Gonoi
T,
Kobayashi
J.
Amphidinins
C-‐F,
amphidinolide
Q
ana-‐
logues
from
marine
dinoflagellate
Amphidinium
sp.
Org
Lett
2014,
16:
5624-‐5627
[7] Kubota
T,
Iwai
T,
Ishiyama
H,
Sakai
K,
Gonoi
T,
Kobayashi
J.
Amphidinin
G,
a
putative
pre-‐
cursor
of
amphidinin
A
from
marine
dinoflagellate
Amphidinium
sp.
Tetrahedron
Lett
2015,
56:
990-‐993
[8] Iwai
T,
Kubota
T,
Kobayashi
J.
Absolute
configuration
of
amphidinin
A.
J
Nat
Prod
2014,
77:
1541-‐1544
P602
Investigation
of
optimal
methods
including
pH-‐zone-‐refining
centrifugal
partition
chromatography
for
the
isolation
of
communesins
from
cultures
of
a
marine-‐derived
Penicillium
expansum
Thuy
T.
P.
Hoang1,
Nicolas
Borie3,
Audrey
Gratia3,
Yves
François
Pouchus1,
Catherine
Roullier1,2,
Jean-‐Hugues
Renault3,
Olivier
Grovel1,2
1
University
of
Nantes,
EA
2160
-‐
Mer
Molécules
Santé,
9
rue
Bias
BP
53508,
44035
Nantes-‐cedex
1
France,
2
Corsaire-‐ThalassOMICS
Metabolomics
Facility,
Biogenouest,
University
of
Nantes,
France,
3
UMR
CNRS
7312,
Université
de
Reims
Champagne-‐Ardenne,
Bât.
18,
Moulin
de
la
Housse,
BP
1039,
51687
Reims,
Cedex
2,
France
Communesins
are
a
rare
family
of
indole
alkaloids
isolated
from
marine
or
terrestrial
fungal
strains
[1].
To
date,
eleven
communesins
(A-‐K)
have
been
described,
which
share
a
common
and
unique
heptacyclic
core
with
the
presence
of
two
vicinal
quaternary
centres
[2].
Recently,
10
new
analogs
have
been
detected
by
HPLC-‐HRMS/MS
as
trace-‐amounts
in
cultures
of
a
P.
expansum
marine-‐derived
strain
[3].
Very
few
biological
data
have
been
described
for
this
class
of
compounds,
but
an
interesting
activity
on
the
cellular
cytoskeleton
has
been
once
described,
making
them
a
putative
new
class
of
microfilament
disrupting
drugs
[4].
In
this
context,
our
laboratory
has
engaged
a
program
for
the
investigation
of
communesin
chemodiversity
and
structure-‐activity
relationships.
The
aim
of
this
study
was:
1-‐
to
obtain
the
major
compounds
communesin
A
and
B
in
sufficient
quantities
for
their
pharmacological
evaluation,
and
2-‐
to
investigate
the
use
of
pH-‐zone-‐refining
Centrifugal
Partition
Chromatography
(CPC)
for
the
isolation
of
minor
communesin
analogs.
Different
methods
of
extraction
and
purification
were
then
tested
and
the
composition
of
the
fractions
obtained
was
established
by
UHPLC-‐ESI-‐MS/MS.
This
led
to
the
development
of
an
optimized
protocol
including
a
solvent
culture
extraction
followed
by
an
indole
alkaloid
concentration
at
pH
1,
allowing
the
selective
recovery
of
communesins
with
a
yield
of
98%.
70
mg
and
52
mg
of
communesin
A
and
B
respectively
were
then
obtained
as
pure
compounds.
The
alkaloidic
fraction
was
subsequently
fractionated
by
pH-‐zone-‐refining
CPC
in
ascending
mode
using
a
biphasic
solvent
system
composed
of
methyl
tert-‐butyl
ether/acetonitrile/water
(4/1/5,
v/v/v).
The
separation
of
more
than
ten
communesins
was
then
performed,
which
were
further
purified.
Then,
pH-‐zone
refining
CPC
was
shown
to
be
an
efficient
separation
tool
for
fungal
alkaloids
isolation,
which
will
allow
to
reveal
the
structural
diversity
of
this
unique
class
of
complex
natural
products.
Acknowledgements:
Authors
acknowledge
the
Vietnam
International
Education
Development
program
for
a
PhD
grant
for
Thuy
T.P.
Hoang
Keywords: Communesin, Penicillium expansum, centrifugal partition chromatography (CPC)
References:
[1] Zuo
Z,
Ma
D.
Synthetic
studies
toward
communesins.
Isr
J
Chem
2011;
51:
434-‐441
[2] Seo
JH,
Artman
GD,
Weinreb
SM.
Synthetic
studies
on
perphoramidine
and
the
communesins:
Construction
of
the
Vicinal
Quaternary
Stereocenters.
J
Org
Chem
2006;
71:
8891-‐8900
[3] Kerzaon
I,
Pouchus
YF,
Monteau
F,
Le
Bizec
B,
Nourrisson
MR,
Biard
JF
and
Grovel
O.
Structural
investigation
and
elucidation
of
new
communesins
from
a
marine-‐derived
Penicillium
expansum
Link
by
liquid
chromatography/electrospray
ionization
mass
spectrometry.
Rapid
Commun
Mass
Spectrom
2009;
23:
3928-‐3938
[4] Ratnayake
AS,
Yoshida
WY,
Moobery
SL,
Hemscheidt
TK.
J
Org
Chem
2001,
66,
8717
(b)
Retraction
of
this
article:
Ratnayake
AS,
Yoshida
WY,
Mooberry
SL,
Hemscheidt
TK.
J
Org
Chem
2003;
68:
1640
P603
Cytotoxic
activity
screening
of
some
Turkish
marine
sponge
spe-‐
cies
V.
Murat
Kutluay1,
Belma
Konuklugil2,
Iclal
Saracoglu1
1
Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Hacettepe
University,
06100,
Ankara,
Turkey,
2
Department
of
Pharmacognosy,
Faculty
of
Pharmacy,
Ankara
University,
06100,
Ankara,
Turkey
Marine
organisms
have
proved
to
be
a
source
of
potent
pharmacologically
active
compounds.
More
than
15.000
marine
products
have
been
isolated
and
tested
during
the
last
23
years,
until
2015.
Sponges
have
been
a
big
source
of
secondary
metabolites
with
a
great
diversity
of
structures
and
bioactivities.
Research
studies
on
these
sources
have
resulted
in
the
identifica-‐
tion
of
various
bioactive
compounds
which
were
tested
for
their
potential
antiinflammatory,
antimalarial,
antitumour,
antiviral,
antibiotic,
antifouling
properties
[1,
2].
Turkish
coastline
totals
almost
about
8400km.
The
sponges
found
in
Turkish
water,
have
not
been
intensively
studied
yet.
During
the
course
of
our
studies
on
Turkish
marine
sponges,
we
aimed
to
screen
cytotoxic
potentials
of
Agelas
oroides
Schmidt,
Ircinia
variabilis
Schmidt,
Dysidea
avara
Schmidt,
Dictyonella
incisa
Schmidt,
Axinella
polypoides
Schmidt,
Aplysina
aerophoba
Nardo.
Materials
were
collected
from
the
Turkish
coastline
of
the
Aegean
Sea
as
well
as
the
Mediter-‐
ranean
Sea
at
depths
varying
from
10-‐30
m.
Sponges
were
cut
into
small
pieces
and
extracted
with
methanol.
Cytotoxic
activity
was
performed
through
HeLa
(human
cervix
epithelial
can-‐
cer
cell)
by
MTT
method
[3].
All
extracts
showed
concentration
dependent
cytotoxic
activity.
IC50
values
were
found
in
a
range
of
159.0−823.0
(μg/mL)
as
given
in
the
table
below.
Dicty-‐
onella
incisa
was
found
to
be
the
most
active
species
among
all
tested
sponges.
Due
to
these
promising
results
the
samples
will
be
tested
on
other
cancer
and
non-‐cancerous
cell
lines
to
detect
selectivity
as
a
continuing
study.
Sponge
species
Location
IC50
(μg/mL)
Agelas
oroides
Kemer,
Antalya
426.8
Ircinia
variabilis
Turgutreis,
Mugla
287.9
Dysidea
avara
Ayvalik,
Balikesir
214.2
Dictyonella
incisa
Seferihisar,
Izmir
159.0
Axinella
polypoides
Fethiye,
Mugla
823.0
Aplysina
aerophoba
Ayvalik,
Balikesir
376.1
Acknowledgements:
Authors
are
thankful
to
biologist
Bulent
Gozcelioglu,
PhD
for
collecting
sponge
sam-‐
ples.
This
work
was
partially
assisted
by
FP-‐7
IRSES
BACT
295226.
Keywords:
Marine
sponges,
cytotoxic
activity,
HeLa
cell
line,
MTT
method
References:
[1] Blunt
JW,
Copp
BR,
Munro
MHG,
Northcote
PT,
Prinsep
MR.
Marine
natural
products.
Nat
Prod
Rep
2002;
20:
1-‐48
[2] Blunt
JW,
Copp
BR,
Keyzers
RA,
Munro
MHG,
Prinsep
MR.
Marine
natural
products.
Nat
Prod
Rep
2013;
30:
237-‐323
[3] Saracoglu
I,
Inoue
M,
Calis
I,
Ogihara
Y.
Studies
on
constituents
with
cytotoxic
and
cyto-‐
static
activity
of
two
Turkish
medicinal
plants
Phlomis
armeniaca
and
Scutellaria
salviifo-‐
lia,
Biol
Pharm
Bull
1995;
18:
1396-‐1400
P604
Isolation
and
structure
elucidation
of
new
alkaloids
from
the
bryozoan
Flustra
foliacea
Xiaxia
Di1,
Shuqi
Wang2,
Eydis
Einarsdottir1,
Elin
S.
Olafsdottir1,
Sesselja
Omarsdottir1
1
Faculty
of
Pharmaceutical
Sciences,
University
of
Iceland,
Hagi,
Hofsvallagata
53,
IS-‐107
Reykjavik,
Ice-‐
land,
2
Faculty
of
Pharmaceutical
Science,
University
of
Shandong,
44
West
Wenhua
Road,
250012
Jinan,
China
Flustra
folicea
is
an
abundant
bryozoan
in
various
parts
of
the
North
Sea
[1].Chemical
investi-‐
gations
of
the
F.
foliaca
have
resulted
in
a
number
of
unique
brominated
pyrrolo[2,3-‐b]indole
alkaloids
with
isoprenyl
substituents
at
various
positions.
These
alkaloids
have
been
shown
to
have
antibacterial
[1],
nonspecific
voltage-‐sensitive
potassium
channel
blocking
[2]
and
sub-‐
type-‐specific
nicotinic
acetylcholine
receptor
blocking
activities
[3].
The
aim
of
the
study
was
to
perform
a
chemical
investigation
of
the
secondary
metabolites
of
F.
folicea
collected
in
Icelandic
waters
using
UPLC-‐qTOF
MS/MS
analysis,
followed
by
isolation
and
structure
elucidation
of
new
compounds.
Dichloromethane:methanol
extract
of
the
bryo-‐
zoan
was
prepared
and
the
profile
of
secondary
metabolites
analysed
by
UPLC-‐qTOF-‐MS/MS
and
MassLynx.
The
data
indicated
the
presence
of
both
previously
known
as
well
as
new
compounds.
Therefore,
the
extract
was
further
separated
into
fractions
using
various
chroma-‐
tographic
methods
(e.g.
CC,
VLC,
SPE
and
preparative
HPLC)
followed
by
TLC,
1H-‐NMR
and
UPLC-‐MS/MS.
Structure
elucidation
was
established
on
the
basis
of
extensive
spectroscopic
analysis
(NMR,
MS,
UV,
and
IR).
The
stereochemistry
of
the
new
compounds
was
determined
by
NOESY,
proton-‐proton
NMR
coupling
constants,
and
CD
spectra.
In
addition
to
previously
known
compounds,
this
study
revealed
nine
new
brominated
alka-‐
loids,
including
one
rearranged
prenylated
oxindole
alkaloid
(1),
two
prenylated
oxindole
alkaloids
(2−3),
and
six
prenylated
pyrrolo[2,3-‐b]indole-‐
triatomic
heterocyclic
alkaloids
(4−9)
and
two
new
imidazole
alkaloids
(10−11).
The
new
F.
foliacea
alkaloids
will
be
virtually
screened
for
activity
using
an
in
silico
α7
nAChR
model
and
positive
hits
subsequently
tested
in
vitro
for
nAChR
potentiation.
N OH
O
NH
Br N O
NH N
Br N O Br N O
H H Br N R
2 O
1 3
4 R= H
N OH 5 R= OH
R 6 R= OCH3
N N
N O
Br R Br N N
H O
O O
N
H
7 R= H 9 10 R= H
8 R= OH 11 R= CH3
Acknowledgements:
Doctoral
grant
from
the
University
of
Iceland
Research
Fund
Keywords: Flustra folicea, bryozoan, brominated alkaloids, imidazole alkaloids
References:
[1] Peters L, Konig GM, Terlau H, Wright AD. Four new bromo-‐tryptamine derivatives from
the marine bryozoan Flustra foliacea. J Nat Prod 2002; 65: 1633-‐1637
[2] Hodgson JW, Mitchell MO, Thomas ML, Waters KF, Powell D. Deoxypseudophrynaminol: a
novel antibacterial alkaloid. Bioorg Med Chem Lett 1995; 5: 2527-‐2528
[3] Peters L, Wright AD, Kehraus S, Gündisch D, Tilotta MC, König GM. Prenylated indole alka-‐
loids from Flustra foliacea with subtype specific binding on NAChRs. Planta Med. 2004;
70: 883-‐886
P605
Essential
oil
compostion
and
anticandidal
activity
of
Hypericum
elongatum
L.
subsp.
elongatum
Yavuz
Bülent
Köse1,
Gökalp
İşcan2,
Mine
Kürkçüoğlu2,
Sevim
Küçük1
1
Department
of
Pharmaceutical
Botany,
Anadolu
University,
Faculty
of
Pharmacy,
26470,
Eskişehir,
Tur-‐
key; 2 Department of Pharmacognosy, Anadolu University, Faculty of Pharmacy, 26470, Eskişehir, Turkey
The
genus
Hypericum
L.
belonging
to
the
Hypericaceae
family,
has
about
500
species
around
the
world
[1].
Flora
of
Turkey
has
96
species
of
Hypericum
and
46
of
them
are
endemic
[2].
Hypericum
species
are
used
as
sedatives,
antiseptics
and
antispasmodics
in
Turkish
folk
medi-‐
cine
[3].
Aerial
parts
of
Hypericum
elongatum
L.
subsp.
elongatum
was
hydrodistilled
for
3
h
using
a
Clevenger-‐type
apparatus
to
produce
a
small
amount
of
essential
oil
which
was
trapped
in
n-‐hexane.
Oils
were
analysed
by
gas
chromatography-‐mass
spectrometry
(GC-‐MS).
Main
constituents
of
the
oil
were
found
as
α−Pinene
(37.4%),
(E)-‐β-‐ocimene
(23.6%)
and
β-‐
pinene
(10%).
The
essential
oil
was
evaluated
for
its
anticandidal
activity
against
ten
Candida
strains
by
using
partly
modified
CLSI
M27-‐A2
broth
microdilution
method.
The
oil
demon-‐
strated
varied
MIC
results
against
tested
strains
between
31
to
1000µg/mL.
Remarkably,
min-‐
imum
inhibitory
concentration
(MIC)
was
31µg/mL
against
C.
tropicalis.
Keywords: Hypericum, anticandidal activity, essential oil
References:
[1] Nürk
NM,
Madrinán
S,
Carine
MA,
Chase
MW,
Blattner
FR.
Molecular
phylogenetics
and
morphological
evolution
of
St.
John’s
wort
(Hypericum;
Hypericaceae).
Mol
Phylogenet
Evol
2013;
66:
1–16
[2] Güner
A,
Aslan
S,
Ekim
T,
Vural
M,
Babaç
MT.
List
of
Turkish
Flora
(Vascular
Plants).
Publi-‐
cation
of
Nezahat
Gökyiğit
Botanical
Garden
and
Flora
Research
Foundation.
2012
[3] Baytop
T.
Therapy
with
Medicinal
Plants
in
Turkey.
Istanbul
Univ.
Press,˙Istanbul,
Turkey.
1999.
P606
Anti-‐diabetic
and
anti-‐inflammatory
activities
of
the
edible
red
alga
Gracilariopsis
chorda
Yukyoung
Jeon1,
Youn
Hee
Nam2,
Tong
Ho
Kang2,
Jong
Hwan
Kwak1
1
School
of
Pharmacy,
Sungkyunkwan
University,
Suwon,
Gyeonggi-‐do
440-‐746,
Republic
of
Korea,
2
De-‐
partment
of
Oriental
Medicinal
Materials
and
Processing,
College
of
Life
Sciences,
Kyung
Hee
University,
Gyeonggi-‐do
449-‐701,
Republic
of
Korea
Gracilariopsis
chorda
(Holmes)
Ohmi
is
an
edible
perennial
red
alga
in
the
family
Gracilaria-‐
ceae
and
is
distributed
in
the
coastal
area
of
Gangneung,
and
Jeju
islands
in
Korea
[1].
Previ-‐
ous
studies
on
G.
chorda,
have
investigated
anti-‐inflammatory
activity
and
neuroprotective
effects
[2].
In
continuation
of
our
search
for
bioactive
natural
products,
we
have
found
that
the
EtOH
extract
of
G.
chorda
has
antihyperglycemic
activity
in
the
zebrafish
model
for
type
1
diabetes,
and
anti-‐inflammatory
activity.
The
zebrafish
model
for
type
1
diabetes
was
induced
by
alloxan,
which
causes
pancreatic
β-‐cell
necrosis
[3].
The
EtOH
extract
was
consecutively
partitioned
with
hexane,
CH2Cl2,
EtOAc
and
n-‐BuOH
to
give
five
fractions.
Anti-‐diabetic
activi-‐
ty
of
the
solvent
fractions
was
evaluated
by
using
a
zebrafish
model.
Following
alloxan
treat-‐
ment,
pancreatic
islet
size
and
fluorescence
intensity
were
measured.
Among
the
fractions,
the
CH2Cl2,
n-‐BuOH
and
H2O
fractions
revealed
potent
anti-‐diabetic
activity
for
type
1
at
a
concentration
of100
μg/ml.
We
also
evaluated
the
levels
of
inflammatory
mediators
in
LPS-‐
stimulated
RAW
264.7
macrophages,
and
TNF-‐α-‐induced
VCAM-‐1
expression
in
MOVAS-‐1
vascular
smooth
muscle
cells.
The
hexane
fraction
and
hexane
and
CH2Cl2
fractions
signifi-‐
cantly
decreased
IL-‐6
and
TNF-‐α
production,
respectively.
The
VCAM-‐1
expression
was
inhib-‐
ited
significantly
by
hexane,
CH2Cl2
and
H2O
fractions.
Acknowledgements:
This
work
was
supported
by
a
grant
from
Marine
Biotechnology
Program
(PJT200620,
Genome
Analysis
of
Marine
Organisms
and
Development
of
Functional
Applications)
funded
by
Ministry
of
Oceans
and
Fisheries.
Keywords: Gracilariopsis chorda, type 1 diabetes, zebrafish, anti-‐inflammation, VCAM-‐1
References:
[1] Boo
SM,
Ko
YD.
Marine
Plants
from
Korea.
Seoul:
Junghaeng-‐Sa,
2012;
188
[2] Mohibbullah
M,
Hannan
MA,
Choi
J-‐Y,
Bhuiyan
MMH,
Hong
Y-‐K,
Choi
J-‐S,
Choi
IS,
Moon
IS.
The
edible
marine
alga
Gracilariopsis
chorda
alleviates
hypoxia/reoxygenation-‐induced
oxidative
stress
in
cultured
hippocampal
neurons.
J
Med
Food
2015;
18:
960-‐971
[3] Desgraz
R,
Bonal
C,
Herrera
PL.
β-‐Cell
regeneration:
the
pancreatic
intrinsic
faculty.
Trends
Endocrinol
Met
2011;
22:
34-‐43
P607
Halogenated
alkaloids
from
Taiwanese
zoanthid
Zoanthus
kuro-‐
shio
Yu-‐Ming
Hsu1,
I-‐Wen
Lo1,
Yang-‐Chang
Wu1,2,
Fang-‐Rong
Chang1,
Yuan-‐Bing
Cheng1
1
Graduate
Institute
of
Natural
Products,
College
of
Pharmacy,
Kaohsiung
Medical
University,
Kaohsiung,
Taiwan,
2
School
of
Pharmacy,
College
of
Pharmacy,
China
Medical
University,
Taichung
40402,
Taiwan
Marine
invertebrates
are
proven
to
be
rich
sources
of
bioactive
natural
products,
which
are
functionalized
as
chemical
defense
substances
against
other
reef
creatures.
Diverse
secondary
metabolites
are
secreted
directly
by
themselves
or
accumulated
through
the
food
chain.
Tai-‐
wan
connects
Japanese
and
Philippine
islands
and
is
surrounded
by
Kuroshio
and
Oyashio
Currents
with
high
biodiversity.
Zoanthid,
a
sessile
benthos
with
two
rows
of
tentacles
and
appealing
colors,
is
one
of
the
marine
organisms
widely
distributed
in
subtropical
or
tropical
areas.
Zoanthus
kuroshio,
the
most
dominant
species
in
western
coast
of
Taiwan,
having
a
flu-‐
orescent-‐pink
oral
disc
and
brown
tentacles,
was
collected
and
investigated
for
its
secondary
metabolites.
Previously,
a
series
of
alkaloids
were
identified
by
our
research
group
[1].
In
our
further
chemical
investigation,
four
new
alkaloids
named
kuroshines
H－K
(1－4),
together
with
four
known
compounds,
kuroshines
A
and
D
(5
and
6),
zoanthenamide
(7),
and
28-‐
dexozoanthenmide
(8)
were
isolated
from
the
methanolic
extract
of
Z.
kuroshio.
All
of
these
marine
alkaloids
possess
a
densely
functionalized
octacyclic
ring
system
on
the
basis
of
the
zoanthamine
framework.
In
particular,
kuroshines
H
and
I
(1
and
2)
were
the
first
halogenat-‐
ed
zoanthamine-‐type
alkaloids
in
nature.
The
structures
of
these
compounds
were
identified
by
the
interpretation
of
spectroscopic
methods,
especially
2D
NMR
technologies
(COSY,
HMQC,
HMBC,
and
NOESY).
O O O O
H O H H H
OH O O O
H O H O H O H O
H O H O H O H O
R2 O
O O
N N N N
R1 R2
O O O O
R1 R1
1 R1 =I 2 R 1=H R2 =Cl 3 R 1=H R 2=OH 8
7 R1 =H 5 R 1=H R2 =OH 4 R 1=OMe R 2=H
6 R 1=OH R 2=OH
Acknowledgements: The authors thank the grants from ministry of science and technology of Taiwan
(MOST103-‐2628-‐B-‐037-‐001-‐MY3 award to Y.-‐B C). The authors also thank the support from Kaohsiung
medical university (Aim for the Top Universities Grant, grant No. KMU-‐TP104H02)
Keywords: Zoanthid, Zoanthus kuroshio, halogenated alkaloid
References:
[1] Cheng
Y-‐B,
Lan
C-‐C,
Liu
W-‐C,
Lai
W-‐C,
Tsai
Y-‐C,
Chiang
M-‐Y,
Wu
Y-‐C,
Chang
F-‐R.
Ku-‐
roshines
A
and
B,
new
alkaloids
from
Zoanthus
kuroshio.
Tetrahedron
Lett
2014;
55:
5369-‐5372
P608
Two
new
polyhydroxylated
compounds
from
the
fungus
Malbranchea
circinata
with
α-‐glucosidase
inhibitory
properties
Abraham Madariaga-‐Mazón, Fernando Cedillo-‐Torres, Laura Flores-‐Bocanegra, Rachel Mata
Departamento
de
Farmacia,
Universidad
Nacional
Autónoma
de
México,
Ciudad
de
México,
04510,
Méxi-‐
co
Type
2
diabetes
mellitus
(TII-‐DM)
is
one
of
the
most
important
public
health
problems
in
all
nations
in
the
21st
century,
since
the
disease
leads
to
serious
complications
resulting
in
in-‐
creasing
disability
and
reduction
of
life
expectancy
[1].
Although
metformin
remains
the
most
important
oral
agent
for
the
initial
management
of
TII-‐DM,
there
are
increasing
numbers
of
second-‐
and
third-‐line
oral
antidiabetical
agents.
These
include
sulfonylureas,
thiazolidinedi-‐
ones,
incretin-‐based
therapies,
and
α-‐glucosidase
inhibitors
[2].
Since
the
fungal
kingdom
represents
a
valuable
source
of
α-‐glucosidase
inhibitors
[3]
we
decided
to
investigate
Malbranchea
circinata
(ATCC
34526).
Bioactive-‐guided
fractionation
of
a
fungal
extract
from
the
culture
medium
(rice)
led
to
the
isolation
of
two
new
compounds,
a
prenylated
xanthone
(1)
[1,4,8-‐trihydroxy-‐3-‐methyl-‐5-‐(3-‐methylbut-‐2-‐enyl)-‐9H-‐xanthen-‐9-‐one]
and
a
glycosylated
anthraquinone
(2)
[3-‐O-‐(2,3-‐di-‐O-‐acetyl-‐α-‐D-‐ribofuranosyl)-‐1,4,8-‐trihydroxy-‐6-‐methyl-‐9,10-‐
dioxo-‐9,10-‐dyhydroantrhacene].
Conventional
NMR
and
HRESI-‐MS
methods
permitted
their
structural
elucidation.
Compounds
1
and
2
inhibited
the
α-‐glucosidases
enzymes
separated
from
rat
intestinal
acetone
powder
when
tested
with
a
well-‐known
spectrophotometric
assay.
The
half-‐maximal
inhibitory
concentration
(IC50)
values
calculated
were
1.19
±
0.018
mM
and
0.47
±
0.0132
mM,
respectively.
The
results
were
compared
with
those
obtained
for
acarbose
(0.36
±
0.065
mM)
used
as
positive
control.
1 2
Acknowledgements:
This
work
was
supported
by
a
grant
from
CONACyT
grant
219765
Keywords:
Malbranchea
circinata,
anthraquinone,
xanthone,
alpha-‐glucosidase
inhibitors.
References:
[1]
International
Federation
of
Diabetes.
Diabetes
Atlas
IFD,
6th
ed.;
International
Federation
of
Diabetes:
Brussels,
Belgium,
2013
[2]
Israili
HZ.
Advances
in
the
treatment
of
type
2
diabetes
mellitus.
Am
J
Ther
2011;
18:
117–152
[3]
Rivera-‐Chávez
J,
Figueroa
M,
Gonzalez
MC,
Glenn
AE,
Mata
R.
α-‐Glucosidase
inhibitors
from
a
Xylaria
feejeensis
associated
with
Hintonia
latiflora.
J
Nat
Prod
2015;
78:
730−735
[4]
Oki
T,
Matsui
T,
Osajima
Y.
Inhibitory
effect
of
α-‐glucosidase
inhibitors
varies
according
to
its
origin.
J
Agric
Food
Chem
1999;
47:
550–553
P609
Protective
effect
of
oyster
hydrolysate
peptide
in
alcohol
induced
alcoholic
fatty
liver
in
SD-‐rats.
Jae-‐Hyuk
Byun1,
Yeung-‐Joon
Choi3,
Se-‐Young
Choung
1,2
1Department
of
Life
and
Nanopharmaceutical
Sciences,
College
of
Pharmacy,
Kyung
Hee
University,
Hoegi-‐dong,
Dongdaemun-‐gu,
Seoul
130-‐701,
Republic
of
Korea,
2
Department
of
Preventive
Pharmacy
and
Toxicology,
College
of
Pharmacy,
Kyung
Hee
University,
Hoegi-‐dong,
Dongdaemun-‐gu,
Seoul
130-‐701,
Republic
of
Korea,
3Department
of
Seafood
Science
and
Technology/Institute
of
Marine
Industry,
Gyeong-‐
sang
National
University,
Gyeongnam
650-‐160,
Republic
of
Korea
In
the
present
study,
protective
effects
of
oyster
hydrolysate
peptide
(TGPN)
against
alcoholic
liver
were
investigated
in
Sprague-‐Dawley
rats.
5
week
old
Sprague-‐Dawley
male
rats
were
divided
into
two
groups
and
fed
Lieber-‐DeCarli
diet
containing
5%
(w/v)
alcohol
(ED)
or
an
isocaloric
amount
of
dextrin-‐maltose
for
the
controls
(ND)
for
4
weeks
[1],
then
ED
were
treated
with
TGPN
(50,
100,
or
200
mg/kg)
or
silymarin
(100mg/kg)
for
another
4
weeks.
The
TGPN
supplementation
significantly
lowered
serum
lipid
levels
(TG,
TC,
FFA),
hepatic
lipid
levels
(TG,
TC,
FFA),
liver-‐index
and
hepatotoxicity
(ALT,
AST)
in
ED-‐induced
rats.
Histo-‐
logical
analysis
indicated
that
the
TGPN-‐treated
group
showed
lowered
number
of
lipid
drop-‐
lets,
reduced
size
of
adipocytes
and
reduced
hepatic
lipid
amount
compared
to
the
ED
group.
To
understand
the
molecular
mechanism
of
TGPN,
the
levels
of
genes
involved
in
alcoholic
fatty
liver
were
measured.
The
expression
of
fatty
acid
oxidation
and
thermogenesis-‐related
genes
(PPAR-‐α,
CPT-‐1)
of
ED-‐induced
rats
[2]
were
increased
by
TGPN.
On
the
other
hand,
TGPN
treatment
resulted
in
decreased
expression
levels
of
lipogenesis-‐related
genes
such
as
SREBP-‐1c,
FAS,
and
SCD1.
These
results
suggest
that
TGPN
may
have
ability
to
ameliorate
al-‐
coholic
fatty
liver
and
hepatic
protection.
Acknowledgements:
This
study
was
supported
by
the
Korea
Institute
of
Marine
Science
&
Technology
promotion.
Keywords: Oyster hydrolysate, alcoholic fatty liver, lipid metabolism, SD-‐rat
References:
[1] Lieber
CS,
Decarli
LM.
Animal
models
of
chronic
ethanol
toxicity.
Methods
Enzym
1994;
233:
585–594.
[2] Chang
CJ,
Tzeng
TF,
Liou
SS,
Chang
YS,
Liu
IM.
Kaempferol
regulates
the
lipid-‐profile
in
high-‐fat
diet-‐fed
rats
through
an
increase
in
hepatic
PPARα
levels.
Planta
Med
2011;
77:
1876-‐1882.
P610
Halogenated
metabolites
from
Irish
Osmundea
spp.
–
a
three
course
meal
for
Aplysia
sp.
Sylvia
Soldatou1,
2,
Ryan
M.
Young1,
2,
Svenja
Heesch3,
4,
Bill
J.
Baker1,
2
1School
of
Chemistry,
National
University
of
Ireland,
Galway,
Ireland,
2Department
of
Chemistry
and
Cen-‐
ter
for
Drug
Discovery
and
Innovation
(CDDI),
University
of
South
Florida,
Tampa,
FL
33620,
USA,
3Irish
Seaweed
Research
Group,
Ryan
Institute
for
Environmental,
Marine
and
Energy
Research,
National
Uni-‐
versity
of
Ireland,
Galway,
Ireland,
4Laboratoy
of
Integrative
Biology
of
Marine
Models,
Algal
Genetics
Research
Group,
Station
Biologique
de
Roscoff,
Roscoff,
France
Ireland’s
marine
territory
is
ten
times
bigger
than
its
landmass
representing
one
of
the
most
biodiverse
and
species-‐rich
habitats
in
Europe
[1].
With
few
studies
conducted
in
the
NE
At-‐
lantic
region
from
a
biodiscovery
point
of
view,
Irish
waters
are
a
great
source
of
potential
new
and
unexplored
chemistry.
From
the
10.000
algal
species
existing
worldwide
Ireland
hosts
~600
[2].
Red
algae
produce
halogenated
terpenes
and
acetogenins
for
chemical
defence
and
ecological
purposes.
Therefore
they
have
been
studied
as
a
source
of
bioactive
metabo-‐
lites
and
these
compounds
have
been
also
examined
from
an
ecological
point
of
view
[3,4].
Four
red
algal
species
from
the
Osmundea
genus,
have
been
described
from
Irish
waters
with
O.
pinnatifida
and
O.
osmunda
being
the
most
common.
Thus,
this
project
is
focusing
on
the
isolation
and
characterisation
of
secondary
metabolites
from
Osmundea
spp.
collected
from
the
Western
Irish
shore
in
Co.
Galway.
Moreover,
a
sea
hare,
Aplysia
sp.,
has
been
found
to
be
associated
with
Osmundea
spp.
The
goal
of
this
research
is
to
compare
the
chemistry
of
the
two
marine
organisms
and
study
the
ecological
relationship
between
the
algae
and
the
sea
hares.
Particularly,
the
aim
is
to
determine
whether
the
sea
hares
are
sequestering
com-‐
pounds
from
the
algae
or
they
are
using
the
seaweed
as
a
shield
from
environmental
challen-‐
ges.
To
probe
the
ability
of
the
sea
hares
to
sequester
algal
chemistry
we
analysed
sea
hares
freshly
collected
from
their
Osmundea
habitat
and
compared
them
to
a
similar
collection
that
was
first
isolated
in
an
aquarium
where
they
were
allowed
to
clear
their
guts
during
a
week
of
starvation.
The
sea
hare
and
algal
samples
were
extracted
in
organic
solvents
and
their
ext-‐
racts
were
subject
to
NMR-‐guided
isolation
of
secondary
metabolites
using
MPLC
and
HPLC.
The
algal
and
sea
hare
extracts
were
compared
through
metabolomics
analysis
using
LC-‐MS
and
GC-‐MS,
whereas
the
structures
of
pure
compounds
were
determined
by
1D
and
2D
NMR
spectroscopy.
Keywords: red algae, sea hare, GC-‐MS, biodiscovery, metabolomics
Acknowledgements:
This
research
was
carried
out
with
the
support
of
the
Beaufort
Marine
Research
Award
and
the
Marine
Institute,
Oranmore,
Co.
Galway,
Ireland.
We
are
grateful
to
Dr.
Laurent
Calcul
and
Andrew
Shilling
for
helping
with
GC-‐MS
QToF
data
collection
as
well
as
the
State
of
Florida
for
Cen-‐
ter
of
Excellence
funding
of
CDDI
References:
[1] Rae
M,
Folch
H,
Moniz
BJM,
Wolff
WC,
McCormack
PG,
Fabio
Rindi,
F,Johnson
PM.
Marine
bioactivity
in
Irish
waters.
Phytochem
Rev
2013;
12:
555–565.
[2] Guiry
MD,
Guiry
GM.
AlgaeBase
2016.
World-‐wide
electronic
publication,
National
Univer-‐
sity
of
Ireland,
Galway.
http://www.algaebase.org
[3] Cabrita
TM,
Vale
C,
Rauter
PA.
Halogenated
Compounds
from
Marine
Algae.
Mar
Drugs
2010;
8:
2301-‐2317
[4] Crasten
P,
Pohnert
G.
Production
and
role
of
volatile
halogenated
compounds
from
marine
algae.
Nat
Prod
Rep
2011;
28:
186-‐195
P611
α-‐Glucosidase
inhibitors
from
Malbranchea
flavorosea
Abraham
Madariaga-‐Mazón,
Daniela
Rebollar-‐Ramos,
Brisa
Verastegui-‐Omaña,
Laura
Flores-‐
Bocanegra,
Rachel
Mata
Departamento
de
Farmacia,
Universidad
Nacional
Autónoma
de
México,
Ciudad
de
México,
04510,
Méxi-‐
co
Diabetes
mellitus
is
a
metabolic
disorder
characterized
by
chronic
high
blood
glucose
levels,
and
is
estimated
as
one
of
the
most
challenging
health
problems
of
the
21st
century.
The
drugs
available
to
lower
hyperglycemia,
focus
on
increasing
insulin
levels,
improving
sensitiv-‐
ity
to
the
hormone
in
tissues,
or
reducing
the
rate
of
carbohydrate
absorption
from
the
gas-‐
trointestinal
tract.
The
last
group
of
drugs
includes
the
inhibitors
of
α-‐glucosidases.
In
recent
years,
the
search
for
new
α-‐glucosidase
inhibitors
(αGI)
structurally
unique
and
biologically
active
from
natural
sources
has
increased
notably
[1].
As
a
part
of
our
continuing
effort
to
dis-‐
cover
new
α-‐glucosidases
from
natural
sources
[2,
3],
we
have
now
investigated
Malbranchea
flavorosea
(ATCC
34529).
Biodirected
fractionation
of
the
organic
extract
of
M.
flavorosea
grown
in
rice
medium,
led
to
the
isolation
of
three
active
compounds,
identified
as
xylarinol
A
(1)
[4],
xylarinol
B
(2)
[5]
and
massarigeninen
C
(3)
[6].
Their
spectroscopic
and
spectromet-‐
ric
data
was
compared
to
that
previously
reported
on
literature.
1 2 3
The
α-‐glucosidase
inhibitory
activity
was
demonstrated
with
yeast
and
rat
small
intestinal
α-‐
glucosidases
using
a
well-‐known
spectrophotocolorimetric
assay
[6].
The
three
compounds
showed
inhibitory
activity
on
mammalian
α-‐glucosidases
with
half
maximal
inhibitory
con-‐
centrations
(IC50)
ranging
from
1.05
to
1.19
mM,
less
active
than
acarbose
(IC50
0.362
mM)
used
as
positive
control,
but
comparable
to
most
natural
products
reported
as
αGI.
This
is
the
first
report
of
αGI
activity
for
these
natural
products,
and
represents
the
first
chemical
study
of
M.
flavorosea,
contributing
to
the
chemical
knowledge
of
the
genus
Malbranchea.
Acknowledgements: This work was supported by a grant from CONACyT (219765).
Keywords: Malbranchea flavorosea, α-‐glucosidase inhibitors, xylarinol.
References:
[1] Israili
ZH.
Advances
in
the
treatment
of
type
2
diabetes
mellitus.
Am
J
Ther
2011;
18:
117−152
[2] Mata
R,
Cristians
S,
Escandón-‐Rivera
S,
Juárez-‐Reyes
K,
Rivero-‐Cruz
I.
Mexican
antidiabetic
herbs:
Valuable
sources
of
inhibitors
of
α-‐glucosidases.
J
Nat
Prod
2013;
76;
468−483
[3] Rivera-‐Chávez
J,
Figueroa
M,
González
MDC,
Glenn
AE,
Mata
R.
α-‐Glucosidase
inhibitors
from
a
Xylaria
feejeensis
associated
with
Hintonia
latiflora.
J
NatProd.
2015;
78;
730−735
[4] Lee
IK,
Jang
YW,
Kim
YS,
Yu
SH,
Lee
KJ,
Park
SM,
Oh
BT,
Chae
JC,
Yun
BS.
Xylarinols
A
and
B,
two
new
2-‐benzoxepin
derivatives
from
the
fruiting
bodies
of
Xylaria
polymorpha.
J
Anti-‐
biot
2009;
62;
163−165
[5] Mullapudi
V,
Ramana
CP.
The
total
synthesis
and
structural
assignment
of
hexaketide
xylarinol
B
and
its
C1’-‐epimer.
Asian
J
Org
Chem
2016;
5:
417−422
[6] Oh
H,
Swenson
DC,
Gloer
JB,
Shearer
CA.
New
bioactive
rosigenin
analogues
and
aromatic
polyketide
metabolites
from
the
freshwater
aquatic
fungus
Massarina
tunicata.
J
Nat
Prod
2003;
66:
73−79
P612
Statistical
optimization
of
production
and
isolation
of
cytotoxic
compound
from
Nocardia
ignorata
Alba
Noël1,
Gwendoline
Van
Soen1,
Isabelle
Rouaud1,
Solenn
Ferron1,
Eric
Hitti2,3,
Sophie
To-‐
masi1
1
PNSCM,
UMR
CNRS
ISCR
6226,
UFR
Sciences
Pharmaceutiques
et
Biologiques,
Université
Bretagne
Loire,
2
Av.
du
Professeur
Léon
Bernard,
35043
Rennes,
France,
2
INSERM
UMR
1099,
Rennes,
France,
3
Univer-‐
sité
de
Rennes
1,
LTSI,
Rennes,
France.
Lichens
are
symbiotic
organisms
consisting
of
fungi
and
algae
(Chlorophyta
or
Cyanobacte-‐
ria),
and
are
well-‐known
as
rich
sources
of
novel
compounds
[1].
However,
recent
studies
re-‐
vealed
the
presence
of
bacterial
microbiota
associated
with
these
organisms
[2].
These
mi-‐
croorganisms
may
represent
an
underexplored
reservoir
of
novel
species
of
potential
interest
in
the
discovery
of
novel
lead
compounds.
However,
the
biosynthesis
of
original
microbial
compounds
is
influenced
by
cultivation
conditions
such
as
nutrients,
incubation
periods,
pH,
temperature,
etc.
Changing
these
parameters
according
to
the
OSMAC
approach
[3]
(One
Strain
MAny
Compounds)
can
result
in
different
secondary
metabolite
profiles.
Actinobacteria
are
known
for
their
ability
to
produce
compounds
of
clinical
and
pharmaceutical
importance
[4].
Thus,
we
applied
the
OSMAC
approach
to
Nocardia
ignorata,
which
was
isolated
from
Col-‐
lema
auriforme
[5].
The
extracts
obtained
from
various
culture
media
(LB,
TY,
MB,
ISP2)
showed
interesting
cytotoxicity
against
two
different
cell
lines.
A
statistical
optimization
of
the
production
of
active
compound
was
established
using
Tagushi
L9
orthogonal
array
design.
The
four
selected
parameters
were
temperature,
pH,
volume
and
oxygenation
of
the
medium
with
three
different
levels
for
each
parameter.
After
several
days
of
growth
using
a
5L
biore-‐
actor
BioFlo®
115,
the
culture
medium
TY
was
collected
and
extracted
with
XAD-‐7
resin
yielding
two
extracts
(supernatant
and
resin
extracts).
A
bioguided
isolation
process
showed
that
the
cytotoxicity
was
focused
in
the
supernatant
(HaCaT
(Human
keratinocyte
cells):
IC50
=
8.0
±
1.5
µg/mL;
B16
(Murine
melanoma
cells):
IC50
=
4.6
±
1.8
µg/mL).
This
extract
was
then
purified
using
flash
chromatography,
semi-‐preparative
reversed-‐phase
HPLC
using
a
water/CH3CN
gradient.
This
process
led
to
the
isolation
of
an
active
compound
which
is
a
novel
bromide
diketopiperazine
identified
by
NMR
and
mass
spectrometry
data.
Acknowledgements:
Delphine
Parrot
is
acknowledged
for
bacterial
isolation
and
preliminary
assays.
The
“Ligue
contre
le
cancer”
is
also
acknowledged
for
financial
support
for
the
BioFlo®
115
bioreactor.
Keywords: Actinobacteria, Nocardia ignorata, Bioguided fractionation, Tagushi experimental
design, OSMAC approach
References:
[1] Shukla
V,
Joshi
GP,
Rawat
MSM.
Lichens
as
a
potential
natural
source
of
bioactive
compounds:
a
review.
Phytochem
Rev
2010;
9:
303-‐314.
Shrestha
G,
St
Clair
LL.
Lichens:
a
promising
source
of
antibiotic
and
anticancer
drugs.
Phytochem
Rev
2013;
12:
229–244.
[2] González
I,
Ayuso-‐Sacido
A,
Anderson
A,
Genilloud
O.
Actinomycetes
isolated
from
lichens:
Evaluation
of
their
diversity
and
detection
of
biosynthetic
gene
sequences.
FEMS
Micro-‐
biol
Ecol
2005;
54:
401–415.
Cardinale
M,
Puglia
AM,
Grube
M.
Molecular
analysis
of
li-‐
chen-‐associated
bacterial
communities.
FEMS
Microbiol
Ecol
2006;
57:
484–495.
Grube
M,
Cardinale
M,
Vieira
de
Castro
J
Jr,
Müller
H,
Berg
G.
Species-‐specific
structural
and
func-‐
tional
diversity
of
bacterial
communities
in
lichen
symbioses.
Int
Soc
Microb
Ecol
2009;
3:
1105–1115.
[3] Bode
HB,
Bethe
B,
Höfs
R,
Zeeck
A.
Big
effects
from
small
changes:
possible
ways
to
explore
nature's
chemical
diversity.
Chem
Bio
Chem
2002;
3:
619-‐627.
[4] Bérdy
J.
Bioactive
Microbial
Metabolites.
J
Antibiot
2005;
58:
1–26.
[5] Parrot
D,
Antony-‐Babu
S,
Intertaglia
L,
Grube
M,
Tomasi
S,
Suzuki
MT.
Littoral
lichens
as
a
novel
source
of
potentially
bioactive
Actinobacteria.
Sci
Rep
2015;
5:
15839.
P613
Characterization
of
alkaloids
and
carotenoids,
a
defense
cocktail
on
Coccinellidae
eggs’
surface.
Alexandre Maciuk1, Pedro Vásquez-‐Ocmín1, Felipe Ramon-‐Portugal2, Gilles Espinasse2, Erwan
Poupon1, Alexandra Magro2
1 Faculté de Pharmacie, Université Paris Sud, CNRS UMR 8076 BioCIS, Equipe de Chimie des Substances
Naturelles,
Université
Paris-‐Saclay,
5
rue
J-‐B.
Clément
92296,
Chatenay-‐Malabry
Cedex,
France,
2
Labora-‐
toire
Evolution
et
diversité
biologique,
UMR
CNRS
5174,
Université
de
Toulouse
–
ENFA,
2
route
de
Nar-‐
bonne,
31326
Castanet
Tolosane
Cedex,
France
Ladybirds
(Coccinellidae:
Coleoptera)
are
known
to
produce
sophisticated
secondary
metabo-‐
lites,
including
alkaloids,
used
mainly
for
defense.
These
molecules
are
present
in
different
life
stages,
from
eggs
to
adults.
Our
attention
was
drawn
to
the
Calvia
quatuordecimguttata
(L.)
species
eggs,
that
are
covered
with
red
drops.
Behavioral
studies
show
that
this
secretion,
produced
during
egg
laying,
very
effectively
repels
ladybird
predators.
The
minute
amounts
of
secretion
available
are
a
challenge
of
the
chemist.
After
collection
with
SPME
fibers
under
a
stereo
microscope,
secretions
were
analyzed
by
LC-‐UV-‐ESI-‐QToF.
They
contain
calvine
ana-‐
logs
and
carotenoids.
Carotenoids
are
believed
to
be
a
reliable
signal
of
the
toxic
or
distasteful
presence
of
the
calvine
alkaloid.
This
example
shows
how
ecologists
and
chemists
benefit
from
joint
effort
to
explore
chemical
ecology
mysteries.
Keywords: Calvia, calvine, β-‐carotene
References:
[1] Braekman
JC, Charlier
A, Daloze
D, Heilporn
S, Pasteels
J, Plasman
V,
Shaofang
W.
New
Piperidine
Alkaloids
from
Two
Ladybird
Beetles
of
the
Genus
Calvia
(Coccinellidae).
Eur
J
Org
Chem
1999;
1749-‐1755
[2] Ware
RL,
Ramon-‐Portugal
F,
Magro
A,
Ducamp
C,
Hemptinne
JL,
Majerus
MEN.
Chemical
protection
of
Calvia
quatuordecimguttata
eggs
against
intraguild
predation
by
the
inva-‐
sive
ladybird
Harmonia
axyridis.
BioControl
2008
;
53:
189-‐200
P614
Sectorial
land
snail
damage
to
the
lichen
Argopsis
friesiana
could
be
explained
by
metabolite
profiles.
Alice
Gadéa1,2,
Françoise
Le
Dévéhat1,
Anne-‐Cécile
Le
Lamer1,3,
Pierre
Le
Pogam1,
Damien
Ertz4,
Maryvonne
Charrier2
,
Joël
Boustie1
1
Université
de
Rennes
1,
ISCR,
UMR
CNRS
6226
PNSCM,
2
avenue
du
Professeur
Léon
Bernard,
35043
Rennes,
France,
2Université
de
Rennes
1,
UMR
CNRS
6553,
263
avenue
du
Général
Leclerc,
35042
Rennes,
France,
3Université
de
Toulouse,
UPS,
UMR
152
Pharma-‐DEV,
Université
Toulouse
3,
Faculté
des
Sciences
Pharmaceutiques;
35
Chemin
des
Maraîchers,
31062
Toulouse
cedex
9,
France,
4
Botanic
Garden
Meise,
Department
Bryophytes-‐Thallophytes
(BT),
Nieuwelaan
38,
B-‐1860
Meise,
Belgium
Notodiscus
hookeri
is
the
only
native
land
snail
of
the
sub-‐Antarctic
present
in
Crozet
Archi-‐
pelago.
Two
shell
ecophenotypes
co-‐exist
on
Possession
Island
and
feed
exclusively
on
lichens
[1]
that
are
symbiotic
organisms
widely
distributed
in
sub-‐Antarctic
islands.
To
improve
our
understanding
of
lichen-‐snail
trophic
interactions,
the
endemic
tripartite
lichen
Argopsis
frie-‐
siana
was
selected.
The
snail
belonging
to
the
organic
ecophenotype
occurs
sympatrically
with
A.
friesiana.
Profiling
and
quantification
of
the
major
secondary
metabolites
in
the
whole
lichen
were
carried
out
by
LC-‐DAD-‐MS.
Two
main
compounds
were
identified,
the
depside
atranorin
(8.2
mg·g-‐1
DM)
and
the
depsidone
argopsin.
Besides,
a
screening
of
primary
me-‐
tabolites
was
carried
out
to
determine
which
compounds
could
be
useful
for
the
snail
energet-‐
ic
demands.
Glutamate,
glutamine,
gamma-‐aminobutyric
acid
and
alpha-‐alanine
were
the
main
amino
acids
found
in
the
lichen
in
low
concentrations
(0.4
mg·g-‐1
DM).
Carbohydrates,
including
arabitol
(10.2
mg·g-‐1),
mannitol
(4.0.10-‐4
mg·g-‐1)
and
sucrose
(3.0.10-‐4
mg·g-‐1),
were
also
identified.
In
cultivation,
N.
hookeri
snails
were
fed
with
fructified
thalli
of
A.
friesiana.
Snails
(n=20)
ate
the
podetia
cortex
and
phyllocladia
but
they
clearly
avoided
apothecia,
the
reproductive
organs.
This
sectorial
damage
of
the
lichen
by
the
snail
suggested
a
differential
distribution
of
the
metabolites.
Therefore,
a
profiling
of
secondary
compounds
was
performed
by
Direct
Analysis
in
Real
Time
–
Mass
Spectroscopy
(DART-‐MS)
[2]
on
each
morphological
part
of
the
lichen
(phyllocladia,
apothecia,
podetia
and
cephalodia).
The
identification
and
distribution
of
secondary
metabolites,
fatty
acids,
amino
acids
and
carbohydrates
should
help
us
to
determine
whether
metabolites
affect
snail
feeding
choice
and
which
of
them
could
be
involved.
Acknowledgements:
Aurélie
Bernard
and
Corentin
Daugan
are
acknowledged
for
their
technical
assis-‐
tance.
The
authors
are
also
indebted
to
David
Rondeau
for
giving
access
to
DART-‐MS.
This
work
used
analytical
facilities
of
P2M2
plateform
for
primary
metabolites
(amino-‐acids
and
carbohydrates)
anal-‐
yses.
Sub-‐antarctic
field
trip
was
funded
by
l'Institut
Polaire
Paul-‐Émile
Victor,
Plouzané,
France
(IPEV,
programme
136).
Keywords: Sub-‐antarctic lichens, metabolites profiling, DART-‐MS, Notodiscus hookeri, chemi-‐
cal ecology.
References:
[1] Charrier
M,
Marie
A,
Guillaume
D,
Bédouet
L,
Le
Lannic
J,
Roiland
C,
Berland
S,
Pierre
J-‐S,
Le
Floch
M,
Frenot
Y,
Lebouvier
M.
Soil
Calcium
Availability
Influences
Shell
Ecopheno-‐
type
Formation
in
the
Sub-‐Antarctic
Land
Snail,
Notodiscus
hookeri.
PLoS
ONE.
2013,
8(12):
e84527.
[2] Le
Pogam
P,
Legouin
B,
Le
Lamer
A-‐C,
Boustie
J,
Rondeau
D.
Analysis
of
the
cyanolichen
Lichina
pygmaea
metabolites
using
in
situ
DART-‐MS:
from
detection
to
thermochemistry
of
mycosporine
serinol.
J
Mass
Spectrom
2015,
50(3):454–62.
P615
SOD
inhibitors
research
from
endophytics
fungi
extracts
Alix
Poinso1,
Patricia
Vicendo1,
François
Couderc1,
Nicolas
Fabre2,
Guillaume
Marti2,
Marieke
Vansteelandt2,
Mohamed
Haddad2,
Billy
J.
Cabanillas3,
Varravaddheay
Ong-‐Meang1
1 IMRCP, université Paul Sabatier, 118 route de Narbonne, 31062 Toulouse Cedex 09. 2 UMR 152 Pharma
Dev,
Université
de
Toulouse,
IRD,
UPS,
France.
3
Instituto
de
Investigación
de
la
Amazonía
Peruana
(IIAP),
Iquitos-‐Quistococha,
Perú.
Superoxide
Dismutase
(SOD)
inhibitors
are
proven
to
be
very
interesting
leads
to
cancer
treatment
and
cisplatine
resistance
reversion
[1].
The
Nature
is
a
huge
“pot
of
gold”
for
new
molecules
research
and
among
the
living
organisms;
fungal
endophytes
represent
an
interest-‐
ing
renewable
source
of
new
compounds
as
their
chemical
investigation
is
quite
recent
in
comparison
to
other
organisms.
In
this
study
we
investigated
endophytes
isolated
from
Ama-‐
zonian
plants,
cultured
on
Malt-‐Agar
medium
and
extracted
by
EtOAc.
An
in
vitro
enzymatic
test,
reproducible,
fast
and
well
adapted
for
fast-‐screening
research
of
SOD
inhibitors
was
developed
in
our
research
group.
This
method,
based
on
pyrogallol
colorimetric
properties
[2]
allows
to
highlight
both
the
anti-‐oxidant
properties
of
extracts
and
their
ability
to
inhibit
the
SOD.
The
preliminary
results
show
an
anti-‐SOD
activity
in
16
on
57
endophytic
fungi
ex-‐
tracts.
In
order
to
decipher
putative
active
compounds
on
our
SOD
assay
model,
a
dereplica-‐
tion
strategy
based
on
LC-‐HRMS
profiling
was
undertaken.
The
most
active
strain,
bellowing
the
Pestalotiopsis
genus
(20%
inhibition
of
SOD
activity),
was
selected
and
a
LC-‐MS
profile
of
this
sample
was
performed
in
order
to
find
furocoumarins
moieties
since
benzopyranoids
scaffold
are
potential
inhibitors
of
SOD
as
shown
by
few
reports
[3].
A
[M+H]+
ions
at
235.096
compound,
corresponding
with
aloesol,
a
coumarine
derivative
already
identified
in
Pestalo-‐
tiopsis
genus
[4],
was
found
in
our
sample,
as
well
as
a
lot
of
other
furocoumarines
moieties.
We
are
now
targeting
the
isolation
and
the
purification
of
these
compounds,
to
experience
our
hypothesis.
In
conclusion,
we
confirmed
that
the
pyrogallol
test
is
indeed
a
good
indicator
for
the
SOD
activity
of
natural
samples,
because
its
results
match
with
bibliography.
Moreover,
we
found
new
compound(s)
responsible
for
anti-‐SOD
activity,
what
provide
a
huge
hope
for
cancer
and
parasitosis
treatment.
Keywords: SuperOxide Dismutase, endophytes, metabolomic
References:
[1] Papa
L,
Manfredi
G,
Germain
D.
SOD1,
an
unexpected
novel
target
for
cancer
therapy.
Genes
Cancer
2014;
5:15-‐21
[2] Li
X.
Improved
pyrogallol
autoxidation
method:
a
reliable
and
cheap
superoxide
scaveng-‐
ing
assay
suitable
for
all
antioxidants.
J
Agric
Food
Chem
2012;
60:
6418-‐6424
[3] Raja
SB,
Murali
MR,
Roopa
K,
Devaraj
SN.
Imperatorin
a
furocoumarin
inhibits
periplas-‐
mic
Cu-‐Zn
SOD
of
Shigella
dysenteriae
their
by
modulates
its
resistance
towards
phagocy-‐
tosis
during
host
pathogen
interaction.
Biomed
Pharmacother
2011;
65:
560-‐568
[4] Yang
XL,
Huang
L,
Li
HY,
Yang
DF,
Li
ZZ.
Two
new
compounds
from
the
plant
endophytic
fungus
Pestalotiopsis
versicolor.
J
Asian
Nat
Prod
Res
2015;
17:
333-‐337.
P616
Bacterial
identification
through
machine
learning
Andreas
Helfenstein1,
Päivi
Tammela1
1
Division
of
Pharmaceutical
Biosciences,
Faculty
of
Pharmacy,
University
of
Helsinki,
P.O.
Box
56,
FI-‐
00014 Helsinki, Finland
Machine
learning
provides
powerful
and
versatile
tools
for
the
analysis
of
large
and
complex
datasets.
It
has
many
applications
in
pharma-‐
and
biosciences,
including
identification
of
mi-‐
cro-‐organisms.
In
order
to
find
out
which
machine
learning
algorithm
is
best
suited
for
that
purpose,
we
compared
the
performance
of
five
algorithms
–
artificial
neural
networks,
naïve
Bayes,
genetic
algorithm,
random
forests,
and
support
vector
machines
–
in
identifying
micro-‐
organisms
present
in
pure
and
mixed
cultures.
Basis
data
used
for
the
identification
were
ab-‐
sorbance
spectra
of
three
microbial
strains
(Escherichia
coli,
Staphylococcus
aureus,
and
Can-‐
dida
albicans).
The
predictive
power
of
the
different
algorithms
varied
between
43
and
92
%.
Best
results
were
achieved
with
a
random
forest
classifier,
which
identified
92
%
of
the
samples
correctly.
The
possibility
to
use
simple
absorbance
spectra
to
identify
bacteria
in
co-‐culture
can
lead
to
streamlined
processes
in
various
fields
of
microbiology,
for
example
in
antimicrobial
screen-‐
ing.
Acknowledgements: This study was funded by the Academy of Finland (grant no. 277001 and 284477).
Keywords: Machine learning, microbiology, data mining
P617
Isolation
and
characterization
of
halogenated
monoterpenes
in
the
investigation
of
the
ecological
relationship
between
Antarctic
Plocamium
cartilagineum
and
Paradexamine
fissicauda
Andrew
J.
Shilling1,
Jacqueline
L.
von
Salm1,
Ryan
M.
Young1,
Margaret
O.
Amsler2,
Charles
D.
Amsler2,
James
B.
McClintock2,
Bill
J.
Baker1
1
Department
of
Chemistry
and
Center
for
Drug
Discovery
and
Innovation,
University
of
South
Florida,
Tampa,
FL
33620,
USA,
2
Department
of
Biology,
University
of
Alabama
at
Birmingham,
Birmingham,
AL
35294,
USA
Plocamium
cartilagineum
is
a
widely
occurring
red
algal
species
which
plays
a
major
role
in
structuring
many
benthic
marine
ecosystems
found
in
the
shallow
waters
of
Antarctica
[1].
This
rhodophyte
is
known
to
produce
many
polyhalogenated
monoterpenes
in
various
rela-‐
tive
abundances
which
have
been
linked
through
metabolomics
with
site-‐specificity,
presum-‐
ably
driven
by
ecological
interactions
[1,2].
The
cytotoxic
nature
of
these
secondary
metabo-‐
lites
is
thought
to
convey
ecological
relevance
as
feeding
deterrents
to
sympatric
algal
con-‐
sumers
[3,4].
One
remarkable
exception
to
this
trend
is
seen
in
the
amphipod
Paradexamine
fissicauda,
which
has
been
shown
to
consume
P.
cartilagineum
at
a
level
unpalatable
to
many
other
associated
amphipod
species
[5].
Further
investigations
have
detected
halogenated
monoterpenes
in
the
metabolome
of
specimens
fed
P.
cartilagineum,
while
not
in
those
fed
non-‐chemically
defended
algae,
with
the
former
enjoying
a
lower
probability
of
being
eaten
by
fish,
suggesting
P.
fissicauda
is
not
only
tolerating
these
chemical
defenses,
but
sequester-‐
ing
halogenated
monoterpenes
for
its
own
defense
[5].
In
order
to
gain
a
greater
understand-‐
ing
of
this
relationship,
numerous
halogenated
monoterpenes
were
isolated
from
P.
cartilagi-‐
neum
specimens
collected
at
Palmer
Station
in
Antarctica
for
subsequent
field-‐based
feeding
assays
and
further
metabolomics
studies.
Metabolites
which
were
shown
to
exist
in
the
great-‐
est
abundance
within
P.
cartilagineum
extracts
were
targeted
for
subsequent
HPLC
purifica-‐
tion
and
structural
elucidation
through
1D
and
2D
NMR
as
well
as
GC/MS
analysis.
During
the
course
of
this
investigation
several
previously
reported
halogenated
monoterpenes
[6]
were
isolated
and
characterized
while
multiple
new
structures
of
the
same
class
were
also
found.
Acknowledgements:
We
are
grateful
for
financial
support
from
the
NSF
(PLR-‐1341339)
and
the
State
of
Florida
for
Center
of
Excellence
funding
of
CDDI.
We
thank
the
field
team
members
of
S-‐022
and
the
staff
of
Antarctic
Support
Contract
for
logistical
support,
Edwin
Rivera
for
his
assistance
in
obtaining
high
quality
NMR
spectra,
Laurent
Calcul
for
his
assistance
with
mass
spec
and
Patrick
Walther
for
his
assis-‐
tance
in
HPLC
purification.
Keywords: Halogenated monoterpenes, Plocamium cartilagineum, Paradexamine fissicauda,
secondary metabolite, chemical defense, chemical sequestration
References:
[1] Young
RM,
von
Salm
JL,
Amsler
MO,
Lopez-‐Bautista
J,
Amsler
CD,
McClintock
JB,
Baker
BJ.Site-‐specific
variability
in
the
chemical
diversity
of
the
Antarctic
red
alga
Plocamium
cartilagineum.
Mar
Drugs
2013;
11:
2126-‐2139
[2] Kladi
M,
Vagias
C,
Roussis
V.
Volatile
halogenated
metabolites
from
marine
red
algae.
Phy-‐
tochem
Rev
2004;
3:
337-‐363
[3] Amsler
CD,
Iken
K,
McClintock
JB,
Amsler
MO,
Peters
KJ,
Hubbard
JM,
Furrow
FB,
Baker
BJ.
Comprehensive
evaluation
of
the
palatability
and
chemical
defenses
of
subtidal
macroal-‐
gae
from
the
Antarctic
Peninsula.
Mar
Ecol
Prog
Ser
2005;
294:
141-‐159
[4] Ankisetty
S,
Nandiraju
S,
Win
H,
Park
YC,
Amsler
CD,
McClintock
JB,
Baker
JA,
Diyaba-‐
lanage
TK,
Pasaribu
A,
Singh
MP,
Maiese
WM, Walsh
RD,
Zaworotko
MJ,
Baker
BJ.
Chemi-‐
cal
investigation
of
predator-‐deterred
macroalgae
from
the
Antarctic
Peninsula.
J
Nat
Prod
2004;
67:
1295-‐1302
[5] Amsler
MO,
Amsler
CD,
von
Salm
JL,
Aumack
CF,
McClintock
JB,
Young
RM,
Baker
BJ.
Toler-‐
ance
and
sequestration
of
macro
algal
chemical
defenses
by
an
Antarctic
amphipod:
a
‘cheater’
among
mutualists.
Mar
Ecol
Prog
Ser
2013;
490:
79-‐90
[6] Stierle
DB,
Wing
RM.
Sims
JJ.
Polyhalogenated
acyclic
monoterpenes
from
the
red
alga
Plocamium
of
Antarctica.
Tetrahedron
1979;
35:
2855-‐2859
P618
Trichaptum
abietinum
from
British
Columbia
exhibited
anti-‐
proliferative
and
immuno-‐modulatory
activities
Ankush
Barad1,
Sumreen
Javed1,
Chow
H.
Lee2
1,2Biochemistry
and
Molecular
Biology
(Chemistry
Program,
University
of
Northern
British
Columba,
3333
University Way, Prince George British Columbia, Canada, V2M 4E2)
Increasing
mortality
rate
and
case
of
occurrence
makes
cancer
a
huge
public
health
challenge
in
both
developed
and
developing
countries
[1].
Toxicity,
adverse
effects,
and
non-‐specificity
are
some
of
the
major
problems
with
the
current
drugs
used
in
cancer
chemotherapy
[2].
For
centuries,
mushrooms
have
been
used
for
their
medicinal
properties
by
different
cultures
[3].
In
the
past
few
decades,
several
natural
bioactive
compounds
from
mushrooms
have
been
reported
to
exhibit
anti-‐cancer
properties
by
means
of
anti-‐proliferation
(reduced
cell
viabil-‐
ity),
immuno-‐stimulation
(stimulating
production
of
cytokines),
apoptosis
(programmed
cell
death)
and
anti-‐inflammation
(inhibiting
excess
unregulated
cytokine
production)
[3].
This
study
aims
to
investigate
potential
anti-‐cancer
properties
of
a
wood-‐decaying
wild
mush-‐
room,
Trichaptum
abietinum
(Ryvarden
L.,
1972)
[Polyporaceae],
which
is
native
to
British
Columbia.
Fresh
fruiting
bodies
of
T.
abietinum
obtained
from
the
Forest
for
the
World
area
in
Prince
George,
British
Columbia,
were
first
identified
by
DNA
sequencing.
Powdered
T.
abiet-‐
inum
was
subjected
to
a
sequential
extraction
using
85%
methanol,
50%
ethanol,
hot
water
and
5%
sodium
hydroxide.
The
extracts
obtained
were
subjected
to
the
anti-‐proliferative
MTT
assay
on
HeLa
cells.
The
fractions
were
also
subjected
to
ELISA,
using
macrophage
Raw
264.7
cells,
for
assessment
of
immuno-‐stimulation
and
anti-‐inflammatory
activities.
At
5
µg/mL,
the
85%
ethanol
fraction
was
found
to
inhibit
the
production
of
lipopolysaccharide
(LPS)-‐induced
tumor
necrosis
factor
(TNF)-‐alpha
production
from
4042pg/ml
to
180.91pg/ml.
At
5µg/ml,
hot
water
extract
was
found
to
be
immuno-‐stimulant,
inducing
2530.9pg/ml
of
TNF-‐alpha
as
compared
to
3897pg/ml
induced
by
LPS.
The
ethanol
and
hot
water
fraction
exhibited
anti-‐proliferative
activity
at
5µg/mL
by
reducing
cell
viability
to
60%.
Future
research
involves
purification
and
identification
of
the
bioactive
from
85%
etha-‐
nol
and
hot
water
fraction.
Fractions
from
T.
abietinum
Biological
activity
85%
Ethanol
(at
5µg/ml)
1. Exhibits
Immuno-‐inhibition
by
inhibiting
LPS
in-‐
duced
TNF-‐alpha
production
from
4042pg/ml
to
180.91pg/ml
in
Raw
264.7
cells
2. Exhibits
anti-‐proliferative
activity
by
reducing
60%
cell
viability
in
HeLa
cells.
Hot
water
(at
5µg/ml)
1. Exhibits
immuno-‐stimulation
by
inducing
2530.9
pg/ml
TNF-‐alpha
as
compared
to
3897pg/ml
in-‐
duced
by
LPS.
2. Exhibits
anti-‐proliferative
activity
by
reducing
60%
cell
viability
in
HeLa
cells
Acknowledgements:
Dr.
Keith
Egger,
Dr.
Hugues
Massicotte,
Dr.
Kerry
Reimer
and
University
of
Northern
British
Columbia.
Keywords:
Anti-‐proliferative,
immuno-‐modulatory,
Trichaptum
abietinum,
mushroom
References:
[1] De
Martel
C,
Ferlay
J,
Franceschi
S,
Vignat
J,
Bray
F,
Forman
D,
Plummer
M.
Global
burden
of
cancer
attribute
to
infections
in
2008:
a
review
and
synthetic
analysis.
Lancet
Oncol
2012;
13:
607−615
[2] Tipton
MT.
Side
effects
of
cancer
chemotherapy.
Skeel,
R.T.
(ed.)
Handbook
of
Cancer
Chemotherapy.
(7th
Edition).
Philadelphia:
Lippincott
Williams
and
Wilkins.
2007;
27:
615−638
[3] Wasser
SP.
Review
of
Medicinal
Mushroom
Advances:
Good
News
from
Old
Allies.
Herbal-‐
Gram
2002;
56:
28−33
P619
Isolation
of
metabolites
from
epigenetically
modified
mangrove
fungi
for
anti-‐infective
drug
discovery
Anne-‐Claire
D.
Limon1,
Renee
Fleeman2,
Lindsey
N.
Shaw2,
Bill
J.
Baker1
1
Departments
of
Chemistry,
2
Cell
Biology,
Microbiology,
and
Molecular
Biology
and
Center
for
Drug
Dis-‐
covery
and
Innovation,
University
of
South
Florida,
Tampa
Florida
33620
Pathogen
resistance,
appearance
of
new
diseases,
and
lack
of
cure
for
neglected
diseases
are
several
motivations
driving
the
search
for
new
treatments
[1,
2].
The
chemical
survival
mech-‐
anisms
used
by
fungi
offer
a
potential
axis
for
research
to
find
new
drugs.
The
production
of
metabolites
being
directly
related
to
the
expression
of
genetic
information
encoded
in
DNA,
regulating
this
resource
will
enable
producing
of
new
molecules
for
the
purpose
of
drug
dis-‐
covery
progress.
Epigenetic
regulation
is
a
key
mechanism
to
orchestrate
the
expression
or
suppression
of
gene
activity
[3].
Manipulating
these
mechanisms
offers
new
opportunities
to
express
down-‐regulated
secondary
metabolite
genes
and
has
the
potential
to
generate
new
potent
and
novel
metabolites
[3].
Screening
studies
in
our
labs
have
shown
that
fungi
from
Floridian
mangroves,
cultured
on
rice
in
the
presence
of
epigenetic
regulators,
exhibited
activity
against
microbial
agents
such
as
ESKAPE
pathogens,
leishmaniasis
and
Naegleria
fowleri
pathogens.
Through
a
bioassay-‐guided
sequence
compiling
extractions,
partitions,
and
chromatographic
methods,
the
separation
of
a
crude
extract
material
has
shown
potential
new
chemistry.
Col-‐
lected
from
the
Tampa
Bay
area
mangrove
in
Florida,
a
DNMT
treated
fungus
was
active
in
ESKAPE
screening.
After
ethyl
acetate/water
partition,
the
crude
extract
was
fractionated
on
normal
phase
MPLC.
Then,
stages
of
normal
phase
and
reverse
phase
HPLC
purifications
re-‐
sulted
in
the
isolation
of
a
polyketide
compound
with
1μM
activity
against
MRSA.
One
and
two-‐dimensional
NMR
spectroscopy
with
mass
spectrometry
provided
the
data
necessary
to
elucidate
the
structure
and
characterize
the
stereochemistry
involved.
Keywords: Epigenetic modifications, mangrove, fungi, drug discovery, ESKAPE pathogens
References:
[1] Boucher
H
W,
Talbot
G
H,
Bradley
J
S,
Edwards
J
E,
Gilbert
D,
Rice
L
B,
Scheld
M,
Spellberg
B,
Bartlett
J.
Bad
bugs,
no
drugs:
no
ESKAPE!
An
update
from
the
Infectious
Diseases
Society
of
America.
Clin
Infect
Dis
2009;
48:
1-‐12
[2] Fleeman
R,
LaVoi
T
M,
Santos
R
G,
Morales
A,
Nefzi
A,
Welmaker
G
S,
Medina-‐Franco
J
L,
Giu-‐
lianotti
M
A,
Houghten
R
A,
Shaw
L
N.
Combinatorial
libraries
as
a
tool
for
the
discovery
of
novel,
broad-‐spectrum
antibacterial
agents
targeting
the
ESKAPE
pathogens.
J
Med
Chem
2015;
58:
3340-‐3355
[3] Beau
J,
Mahid
N,
Burda
W.
N,
Harrington
L,
Shaw
L.
N,
Mutka
T,
Kyle
D.
E,
Barisic
B,
van
Olphen,
A,
Baker
B.
J.
Epigenetic
tailoring
for
the
production
of
anti-‐infective
cytosporones
from
the
marine
fungus
Leucostoma
persoonii.
Mar
Drugs
2012;
10:
762-‐774
P620
The
new
diketopiperazines
produced
by
marine
algicolous
fun-‐
gus
Penicillium
sp.
KMM
4672
Anton
N.
Yurchenko1,
Olga
F.
Smetanina1
1
Laboratory
of
Chemistry
of
Microbial
Metabolites,
G.B.
Elyakov
Pacific
Institute
of
Bioorganic
Chemistry
Far
Eastern
Branch
of
Russian
Academy
of
Science,
Prospect
100-‐letiya
Vladivostoka,
159,
690022
Vladi-‐
vostok,
Russian
Federation
Marine
fungi
are
a
prolific
source
of
chemically
diverse
biologically
active
metabolites
[1,2].
The
features
of
habitats
of
marine
fungi
make
them
very
promising
sources
of
bioactive
com-‐
pounds
[3].
Recently
cephalosporin
C
was
the
only
marine
fungal
compound
that
used
as
a
medical
drug,
but
today
fingolimod
developed
from
myriocin
is
approved
by
FDA
and
EMA,
and
plinabulin,
the
synthetic
derivative
of
diketopiperizine
phenylahistin,
is
under
phase
III
of
clinical
trials
(according
to
clinicaltrials.gov)
[4,5].
Diketopiperazine
alkaloids
are
produced
by
many
fungal
species.
Many
of
them
show
the
var-‐
ious
bioactivities
such
as
antiviral,
antibiotic,
cancer
cells
growth
inhibition,
anti-‐
inflammatory
and
others
[6,7].
In
our
recent
study
of
fungi
isolated
from
Vietnamese
algae
we
found
that
the
strain
of
Penicillium
sp.
produced
several
alkaloid
types
that
were
rare
for
na-‐
ture.
One
of
them
showed
potent
cytotoxic
effect
against
human
prostate
cells.
Acknowledgements:
The
research
was
supported
by
Russian
Science
Foundation
(Grant
No
14-‐14-‐00030)
Keywords: Marine-‐derived fungus, secondary metabolites, alkaloids, bioactivity, cytotoxicity
References:
[1] Nicoletti
R,
Trincone
A.
Bioactive
compounds
produced
by
strains
of
Penicillium
and
Tala-‐
romyces
of
marine
origin.
Mar
Drugs
2016;
14:
37
[2] Blunt
JW,
Copp
BR,
Keyzers
RA,
Munro
MH,
Prinsep
MR.
Marine
natural
products.
Nat
Prod
Rep
2016;
33:
382-‐431
[3] Ebada
S,
Proksch
P.
Marine-‐derived
fungal
metabolites.
In:
Kim
S-‐K
ed,
Springer
handbook
of
marine
biotechnology.
Berlin:
Springer
Berlin
Heidelberg;
2015:
759-‐78
[4] Schueffler
A,
Anke
T.
Fungal
natural
products
in
research
and
development.
Nat
Prod
Rep
2014;
31:
1425-‐1448
[5] Ji
YT,
Liu
YN,
Liu
ZP.
Tubulin
colchicine
binding
site
inhibitors
as
vascular
disrupting
agents
in
clinical
developments.
Curr
Med
Chem
2015;
22:
1348-‐1360
[6] Borthwick
AD.
2,5-‐diketopiperazines:
Synthesis,
reactions,
medicinal
chemistry,
and
bio-‐
active
natural
products.
Chem
Rev
2012;
112:
3641-‐3716
[7] Huang
R-‐M,
Yi
X-‐X,
Zhou
Y,
Su
X,
Peng
Y,
Gao
C-‐H.
An
update
on
2,5-‐diketopiperazines
from
marine
organisms.
Mar
Drugs
2014;
12:
6213-‐6235
P621
De
novo
metabolite
production
through
co-‐cultivation
of
different
fungal
species
on
solid
media
Antonio
Azzollini1,
Jean-‐Luc
Wolfender1,
Katia
Gindro2
1
School
of
Pharmaceutical
Sciences,
University
of
Geneva,
University
of
Lausanne,
Quai
Ernest-‐Ansermet
30,
CH-‐1211
Geneva
4,
Switzerland;
2
Agroscope,
Institute
for
Plant
Production
Sciences
IPS,
Mycology
and
Biotechnology,
Route
de
Duiller
50,
PO
Box
1012,
1260
Nyon
1,
Switzerland
In
the
field
of
natural
products
research,
finding
sources
of
novel
bioactive
compounds
is
of
primary
importance.
In
this
respect,
microorganisms
have
provided
a
large
number
of
biolog-‐
ically
active
molecules,
but
due
to
continuous
re-‐isolation
of
known
secondary
metabolites
this
source
is
losing
attractiveness.
To
discover
original
microbial
natural
products,
new
strategies
that
switch
on
silent
genes
appear
as
an
interesting
alternative
to
yield
more
struc-‐
turally
diverse
secondary
metabolites.
In
this
regard,
the
use
of
fungal
co-‐cultures
for
the
in-‐
duction
of
new
bioactive
compounds
has
emerged
as
a
promising
field
in
drug
discovery
[1,2].
In
this
work,
a
method
based
on
the
use
of
12-‐well-‐plate
miniaturized
Petri
dishes
(2
cm
di-‐
ameter)
and
compatible
with
UHPLC-‐HRMS
metabolomics
has
been
applied
to
screen
for
me-‐
tabolite
induction
in
co-‐cultures
of
four
fungal
species
(Epicoccum
sp.,
Eutypa
sp.,
Fusarium
sp.,
Aspergillus
sp.)
grown
on
solid
media
[3].
A
miniaturized
multi-‐well
procedure
was
used
as
it
allows
growing
and
analysing
with
high-‐throughput
a
large
number
of
samples
(single-‐
and
co-‐cultures)
necessary
for
statistical
studies.
PCA
(Principal
Component
Analysis)
was
performed
in
order
to
explore
the
data
through
an
unsupervised
approach
and
reveal
metabolome
variation
among
single
culture
and
co-‐cultures.
The
results
of
the
screening
showed
that
these
fungal
species
produced
new
compounds
when
co-‐cultivated
with
another
fungus.
For
example,
de
novo
induced
metabolites
(detected
in
the
co-‐culture
but
not
in
the
single
cultures)
were
revealed
in
the
interaction
of
Fusarium
sp.
vs
Aspergillus
sp.
as
well
as
in
the
interaction
of
Eutypa
sp.
vs
Epicoccum
sp.
and
some
fungal
secondary
metabolites,
like
O-‐
methylmellein,
were
dereplicated.
This
miniaturized
strategy
provided
a
satisfactory
repro-‐
ducibility;
moreover
it
is
generic
and
can
be
applied
to
other
types
of
microorganisms
that
can
grow
on
solid
media
such
as
those
that
are
part
of
the
microbiome.
This
study
demonstrates
the
consistent
induction
of
new
metabolites
through
fungal
co-‐cultures.
Keywords:
Fungal
co-‐culture,
de
novo
induction,
screening,
12-‐well-‐plate
References:
[1] Bertrand
S,
Bohni
N,
Schnee
S,
Schumpp
O,
Gindro
K,
Wolfender
JL.
Metabolite
induction
via
microorganism
co-‐culture:
A
potential
way
to
enhance
chemical
diversity
for
drug
discovery.
Biotechnol
Adv
2014;
32:
1180−1204
[2] Glauser
G,
Gindro
K,
Fringeli
J,
De
Joffrey
JP,
Rudaz
S,
Wolfender
JL.
Differential
analysis
of
mycoalexins
in
confrontation
zones
of
grapevine
fungal
pathogens
by
ultrahigh
pressure
liquid
chromatography/time-‐of-‐flight
mass
spectrometry
and
capillary
nuclear
magnetic
resonance.
J
Agric
Food
Chem
2009;
57:
1127−1134
[3] Bertrand
S,
Azzollini
A,
Schumpp
O,
Bohni
N,
Schrenzel
J,
Monod
M,
Gindro
K,
Wolfender
JL.
Multi-‐well
fungal
co-‐culture
for
de
novo
metabolite-‐induction
in
time-‐series
studies
based
on
untargeted
metabolomics.
Mol
BioSyst
2014;
10:
2289−2298
P622
Cytotoxic
activity
of
endolichenic
fungi
isolated
from
the
lichen
Nephroma
laevigatum.
Aurélie
Lagarde1,
Marion
Millot1,
Patricia
Jargeat2,
Tan-‐Sothéa
Ouk1,
Vincent
Sol1,
Lengo
Mambu1
1
Department
of
Pharmacognosy,
University
of
Limoges,
EA1069,
Laboratory
of
Chemistry
of
Natural
Sub-‐
stances,
2
rue
du
Docteur
Raymond
Marcland,
87025
Limoges,
France,
2
University
of
Paul
Sabatier,
UMR5174,
CNRS-‐UPS-‐ENFA,
Laboratory
Evolution
and
Biological
Diversity,
118
route
de
Narbonne,
31062
Toulouse
Cedex
9,
France
Lichens
are
characterized
by
a
symbiotic
association
between
a
fungus
(mycobiont)
and
cya-‐
nobacteria
and/or
alga
(photobiont).
They
produce
various
biologically
active
compounds
due
to
the
great
diversity
of
their
ecosystems,
and
their
ability
to
adapt
their
metabolism
to
extreme
environmental
conditions.
However,
lichen
resources
are
limited.
The
study
of
en-‐
dolichenic
fungi
is
relatively
recent1,
but
the
discovery
of
many
new
substances
demonstrates
that
they
represent
a
promising
source
of
original
natural
products2.
The
aim
of
this
work
is
to
investigate
endolichenic
fungi
from
Nephroma
laevigatum
in
search
for
new
bioactive
com-‐
pounds
with
a
possible
pharmaceutical
interest.
Endolichenic
fungi
were
cultivated
on
solid
medium.
Identification
of
isolated
fungi
was
apprehended
by
barcoding
(PCR
amplification
of
the
ITS
with
ITS4
and
ITS5
primers3,
sequencing
and
comparison
to
gene
libraries).
The
iso-‐
lated
fungi
belong
to
genus
Nemania
(81%),
Daldinia
(13%),
Peziza
(3%)
or
Coniochaeta
(3%).
Variations
in
morphological
appearance
and
growth
speed
of
colonies
were
observed,
and
metabolic
profiling
of
crude
extracts
showed
that
their
composition
was
dependent
on
species,
strain
and
culture
medium.
In
this
context,
6
strains
were
selected
and
extracted
with
EtOAc.
Bioactivity
screening
has
been
performed
on
these
crude
extracts.
In
vitro
activity
against
bacterial
biofilm
(Staphylococcus
aureus:
ATCC
25923)
was
carried
out
and
cytotoxici-‐
ty
was
evaluated
by
MTT
assay
on
HT-‐29;
HCT116;
PC3
and
DU145
cell
lines.
One
crude
ex-‐
tract
showed
(IC50
<
20
µg/mL)
cytotoxic
activity
against
colorectal
carcinoma
(HCT116)
and
human
prostate
cancer
(DU145)
cell
lines
and
another
was
found
to
be
cytotoxic
for
DU145
cell
line.
The
most
bioactive
fungus
was
identified
as
Nemania
aenae
var
aureolatum
and
the
second
active
as
Nemania
serpens.
All
extracts
were
inactive
against
S.
aureus.
Results
are
promising
in
search
of
new
bioactive
compounds
from
endolichenic
fungi.
Acknowledgements:
Region
Limousin
for
financial
support;
AFL
and
L.
Farou
for
Nephroma
localization
Keywords: Nephroma laevigatum, Endolichenic fungi, MTT assay, cytotoxicity, phytochemi-‐
cals
References:
[1] Suryanarayanan
TS,
Thirunavukkarasu
N,
Hariharan
GN,
Balaji
P.
Occurrence
of
non-‐
obligate
microfungi
inside
lichen
thalli.
Sydowia,
2005;
57:
120-‐130
[2] Zhang
K,
Ren
J,
Ge
M,
Li
L,
Guo
L,
Chen
D,
Che
Y.
Mono-‐
and
bis-‐furanone
derivatives
from
the
endolichenic
fungus
Peziza
sp.
Fitoterapia,
2014,
92;
79-‐84
[3] White
TJ,
Bruns
T,
Lee
S,
Taylor
J.
Amplification
and
direct
sequencing
of
fungal
ribosomal
RNA
genes
for
phylogenetics.
In
PCR
protocols:
a
guide
to
methods
and
applications.
Aca-‐
demic
Press,
1990;
315-‐322
P623
New
bioactive
compounds
from
Fusarium
fujikuroi
Birgit
Arndt1,2,
Slavica
Janevska3,
Isabel
Krug1,
Lucas
Maciel
Mauriz
Marques1,
Bettina
Tudzynski3,
Hans-‐Ulrich
Humpf1,2
1
Institute
of
Food
Chemistry,
Westfälische
Wilhelms-‐Universität
Münster,
Corrensstraße
45,
48149
Mün-‐
ster,
Germany,
2
NRW
Graduate
School
of
Chemistry,
Westfälische
Wilhelms-‐Universität
Münster,
Wil-‐
helm-‐Klemm-‐Str.
10,
48149
Münster,
Germany,
3
Institute
of
Plant
Biology
and
Biotechnology,
West-‐
fälische
Wilhelms-‐Universität
Münster,
Schlossplatz
8,
Münster,
Germany
In
the
field
of
natural
products,
moulds
provide
a
large
spectrum
of
bioactive
compounds
and
are
therefore
important
targets
to
analyse.
One
of
these
moulds
is
Fusarium
fujikuroi,
a
rice
pathogen
causing
the
foolish
seedling
disease
due
to
its
secretion
of
the
secondary
metabo-‐
lites
gibberellic
acids,
a
group
of
highly
bioactive
phytohormones
[1].
Besides
these
isopre-‐
noids,
it
produces
a
broad
range
of
other
interesting
compounds,
e.g.
the
cyclic
tetrapeptide
apicidin
F
which
shows
antimalarial
activity
[2].
The
genome
of
F.
fujikuroi
was
fully
se-‐
quenced,
revealing
the
presence
of
altogether
47
putative
secondary
metabolite
gene
clusters,
most
without
yet
assigned
product
[3].
Besides,
global
regulatory
genes
that
encode
positive
or
negative
regulators
of
secondary
metabolite
gene
clusters
are
under
investigation.
With
the
help
of
genetic
engineering,
overexpression
and
deletion
mutants
of
biosynthetic
genes
and/or
global
regulator
genes
were
generated.
In
this
study,
we
present
several
identified
metabolites,
including
the
products
of
a
dimethylallyl
tryptophan
synthase,
a
polyketide
syn-‐
thase
(PKS)
and
a
PKS-‐non
ribosomal
peptide
hybrid
(see
Fig.
1).
Those
were
identified
by
HPLC-‐UV-‐HRMS
measurements
of
the
created
mutants
in
comparison
to
the
wild
type.
To
en-‐
hance
the
identification
process,
the
secondary
metabolite
profiles
were
analysed
with
the
software
tool
MZmine
2
[4],
resulting
in
the
identification
of
new
or
missing
peaks,
respective-‐
ly.
The
compounds
were
isolated
by
different
methods
and
their
structures
were
elucidated
in
detail
by
NMR
and
HPLC-‐HRMS.
With
this
procedure,
known
secondary
metabolites
[5,6]
as
well
as
new,
yet
unknown
compounds
were
identified,
and
corresponding
gene
clusters
char-‐
acterized
[7].
Acknowledgements:
Financial
support
by
the
DFG
(Project
HU
730/9-‐3)
and
the
NRW
Graduate
School
of
Chemistry
is
gratefully
acknowledged.
Keywords: Fusarium, Genomics, Bikaverin, Biosynthesis, Natural Products, Mass spectrome-‐
try, NMR spectroscopy
References:
[1] Kurosawa
E.
Experimental
studies
on
the
nature
of
the
substance
excreted
by
the
“ba-‐
kanae”
fungus.
Trans
Nat
Hist
Soc
Formosa
1926;
16:
213-‐227
[2] von
Bargen
KW,
Niehaus
E-‐M,
Bergander
K,
Brun
R,
Tudzynski
B,
Humpf
H-‐U.
Structure
elu-‐
cidation
and
antimalarial
activity
of
apicidin
F:
an
apicidin-‐like
compound
produced
by
Fusarium
fujikuroi.
J
Nat
Prod
2013;
76:
2136-‐2140
[3] Wiemann
P,
Sieber
CMK,
von
Bargen
KW,
Studt,
L
Niehaus,
E-‐M
Espino,
JJ
Huß,
K,
Mich-‐
ielse
CB,
Albermann
S,
Wagner
D,
Bergner
S
V,
Connolly
LR,
Fischer
A,
Reuter
G,
Kleigrewe
K,
Bald
T,
Wingfield
BD,
Ophir
R,
Freeman
S,
Hippler
M,
Smith
KM,
Brown
DW,
Proctor
RH,
Münsterkötter
M,
Freitag
M,
Humpf
H-‐U,
Güldener
U,
Tudzynski
B.
Decipher-‐
ing
the
Cryptic
Genome:
Genome-‐wide
analyses
of
the
rice
pathogen
Fusarium
fujikuroi
reveal
complex
regulation
of
secondary
metabolism
and
novel
metabolites.
PLoS
Pathog
2013;
9:
1-‐36
[4] Pluskal
T,
Castillo
S,
Villar-‐Briones
A,
Oresic
M.
MZmine
2:
Modular
framework
for
pro-‐
cessing,
visualizing,
and
analyzing
mass
spectrometry-‐based
molecular
profile
data.
BMC
Bioinformatics
2010;
11:
1-‐11
[5] Schultz,
AW,
Lewis,
CA,
Luzung,
MR,
Baran,
PS,
Moore,
BS.
Functional
characterization
of
the
cyclomarin/cyclomarazine
prenyltransferase
CymD
directs
the
biosynthesis
of
un-‐
natural
cyclic
peptides.
J
Nat
Prod
2010;
73:
373-‐377.
[6] Marfori,
EC,
Kajiyama,
S,
Fukusaki,
E,
Kobayashi,
A.
Trichosetin,
a
novel
tetramic
acid
anti-‐
biotic
produced
in
dual
culture
of
Trichoderma
harzianum
and
Catharanthus
roseus
callus.
Z
Naturforsch
C
2002;
57:
465-‐470.
[7] Arndt
B,
Studt
L,
Wiemann
P,
Osmanov
H,
Kleigrewe
K,
Köhler
J,
Krug
I,
Tudzynski
B,
Humpf
H-‐U.
Genetic
engineering,
high
resolution
mass
spectrometry
and
nuclear
magnetic
reso-‐
nance
spectroscopy
elucidate
the
bikaverin
biosynthetic
pathway
in
Fusarium
fujikuroi.
Fungal
Genet
Biol
2015;
84:
26-‐36
Figure
1:
Structures
of
compounds
presented
in
the
study.
P624
Influence
of
Inonotus
hispidus
on
function
of
human
immune
cells
Carsten
Gründemann1,
Mandy
Arnhold1,
Stefanie
Meier2,
Christian
Bäcker2,
Manuel
Garcia-‐
Käufer1,
Franziska
Grunewald1,
Roman
Huber1,
Ulrike
Lindequist2
1
Center
for
Complementary
Medicine,
Institute
for
Environmental
Health
Sciences
and
Hospital
Infection
Control,
Medical
Center-‐
University
of
Freiburg,
Breisacher
Str.
115B,
79111
Freiburg,
Germany,
2
Institu-‐
te
of
Pharmacy,
Ernst-‐Moritz-‐Arndt-‐University
Greifswald,
F.-‐L.-‐Jahn-‐Str.
17,
17487
Greifswald,
Germany
Inonotus
hispidus
is
commonly
used
as
a
traditional
medicine
in
many
regions
of
China.
In
previous
investigations,
fruit
body
extracts
demonstrated
interesting
immunomodulatory
activity,
whereas
bioactivity-‐guided
fractionation
led
to
isolation
and
structure
elucidation
of
hispolon
and
hispidin.
Since
little
is
known
about
the
activities
of
I.
hispidus
in
human
immunocompetent
cells,
we
analyzed
effects
of
I.hispidus
extracts
(IHE)
and
selected
constituents
of
this
mushroom
on
different
types
of
human
immune
cells
and
investigated
the
potential
of
I.
hispidus
as
a
medic-‐
inal
mushroom.
The
influence
of
endotoxin-‐free
IHE
and
selected
compounds
on
activity
and
maturation
of
human
T
cells,
purified
NK
cells
and
monocyte-‐derived
dendritic
cells
were
independently
analyzed
using
flow
cytometric-‐based
surface
marker
expression.
Cell
division
analysis
of
ac-‐
tivated
T
cells
was
assessed
by
the
membrane-‐permeable
dye
carboxyfluorescein
diacetate
succinimidyl
ester
(CFSE)
and
the
function
of
purified
NK
cells
was
investigated
by
the
degranulation-‐CD107a
assay.
Apoptosis
induction
was
analyzed
by
surface
staining
of
phos-‐
phatidylserine
and
camptothecin
and
cyclosporine
A
were
used
individually
as
controls.
Phy-‐
tochemical
analysis,
using
TLC
chromatograms
and
HPLC
analysis,
was
conducted
to
charac-‐
terize
the
IHE.
IHE
increase
the
activation
status
and
diminish
the
proliferation
capacity
of
activated
human
T
cells
in
the
presence
of
apoptosis.
Further
experiments
showed
that
NK
cell
activity
and
function
was
dose-‐dependently
increased.
Surface
marker
expression
of
monocyte-‐derived
dendritic
cells
demonstrated
that
mushroom
extracts
have
the
power
to
induce
maturation
of
these
cells.
TLC
and
HPLC
analysis
showed
that
the
extracts
contain
hispidin
and
hispolon.
Investigations
using
hispidin
and
hispolon
demonstrate
similar
albeit
non-‐congruent
results
with
extracts
on
measured
parameters.
The
results
indicate
that
extracts
from
I.
hispidus
and
their
constituents’
hispidin
and
hispolon
have
the
capability
to
interfere
with
the
function
of
immune
cells
at
more
than
one
site,
thus
providing
a
rationale
for
their
potential
as
a
medicinal
mushroom.
Keywords: Medicinal mushrooms, Inonotus hispidus, Immunomodulation
P625
Anticancer
active
metabolites
from
soil
bacteria
Cassandra Lew1, Chenxi Zoe1, Sandra Loesgen1
1 Department of Chemistry, Oregon State University, 153 Gilbert Hall Corvallis, Oregon 97331-‐4003, USA
Over
the
time
frame
from
around
the
1940s
to
the
end
of
2014,
175
small
molecules
were
approved
as
anticancer
agents.
Of
those,
49%
were
either
natural
products
or
directly
derived
therefrom
[1].
Thus,
small
molecules
still
represent
a
promising
new
source
for
continued
drug
development
and
discovery.
Soil
bacteria
specifically
are
responsible
for
a
number
of
anticancer
drugs
used
clinically
today,
such
as
doxorubicin
and
mitomycin
c.
Herein,
17
soil
bacteria
isolated
from
Bend,
Oregon
were
tested
for
cytotoxic
activity
against
the
a
colon,
breast,
and
prostate
cancer
model
(cell
lines
HCT-‐116,
MCF-‐7,
PC3).
Each
bacterial
strain
was
identified
through
16S
ribosomal
RNA
sequencing.
Ethyl
acetate
extracts
were
tested
using
MTT
cell
viability
and
LDH
cell
cytotoxicity
assays
to
evaluate
their
relative
anticancer
activi-‐
ty.
The
strain
isolation
procedure
and
taxonomy
is
presented
as
well
as
chemical
screening
and
cytotoxic
evaluation
of
these
newly
isolated
bacterial
strains,
derived
from
Oregonian
high
desert.
Bend"Strains" HCT+116"cells"
120"
100"
80"
%"Cell"Viability"
60"
40"
20"
0"
Samples:)10)μg/mL)
Acknowledgements:
Thanks
to
Elizabeth
Kaweesa
and
Birte
Plitzko
for
help
with
the
cancer
assays.
Thanks
to
Patrick
Videau
for
help
with
bacterial
taxonomy
Keywords: Natural product drug discovery, cancer, soil bacteria
References:
[1] Cragg
G,
Newman,
D.
Natural
products
as
sources
of
new
drugs
from
1981
to
2014.
J
Nat
Prod
2016;
79:
629-‐661
P626
The
discovery
and
evaluation
of
anti-‐parasitic
metabolites
from
filamentous
fungi
Cedric
Pearce1,
Blaise
Darveaux1,
Christopher
Rice2,
Beatrice
Colon3,
Kaitlin
Mettel2,
Kati
Rasanen2,
Dennis
Kyle2,
Bill
Baker4,
Nicholas
Oberlies5
1
Mycosynthetix,
Inc.,
505
Meadowlands
Drive
Suite
103,
Hillsborough,
NC,
27278,
USA,
2
Department
of
Global
Health,
College
of
Public
Health,
University
of
South
Florida,
Tampa,
FL,
33612,
USA,
3
Morsani
College
of
Medicine,
University
of
South
Florida,
Tampa,
FL,
33612,
USA,
4
Department
of
Chemistry,
Uni-‐
versity
of
South
Florida,
Tampa,
FL,
33612,
USA,
5
Department
of
Chemistry
and
Biochemistry,
UNC
Greensboro,
NC,
27412,
USA
Mycosynthetix, Inc. a fungus-‐metabolites research organization that employs an extensive
fungus library containing over 50,000 isolates for the production of biologically-‐active metab-‐
olites for human and animal health, and agriculture, has for some time been focused on anti-‐
parasitic lead discovery. We have established validated whole organism assays to determine
activity of fungal metabolites against Haemonchus contortus, Cooperia onchophora, Strong-‐
lyoides stercoralis, Brugia pahangi(Bp), Brugia malayi and Dirofilaria immitis microfilaria, and
Aedes aegypti larvae. Several compounds not only had high potency against BpMf but also
caused high mortality within 24 hours. A small library of carefully selected purified fungal
metabolites was tested in the BpMf assay at 50 ppm, and a number of compounds were fully
active at 3-‐6 μM with the more potent showing nanomolar activity. Encouraged by this and
our joint (CP, DK, BB) earlier work on antimalarial discovery, we initiated a project to find
fungal metabolites active against Naegleria fowleri and Acanthamoeba spp., Free–Living
Amoeba (FLA), that can cause deadly brain diseases called Primary Amoebic Meningoenceph-‐
alitis (PAM) and Granulomatous Amoebic Encephalitis (GAE). Since the first documented in-‐
fection in 1965, 146 PAM cases have been reported in the U.S., with only 3 treated survivors
(98% fatality). Between 1955 and 2012, 63 Acanthamoeba non-‐keratitis cases were docu-‐
mented with 14 treated patients surviving (78% fatality). The major problem for FLA infec-‐
tions is the lack of effective therapeutics discovered specifically for these parasites. The aim of
this study was to employ a validated in vitro HTS method using FLA to screen our large
(30,000) extract libraries. Positive samples were observed between 0.1 and 1% of those test-‐
ed, depending on concentration and included extracts showing minimal cytotoxicity.
Research Supported by: National Institute of Allergy and Infectious Diseases (1R43AI100316-‐1A1, R43
AI118075-‐01A1, R21AI103664 and R21AI119787) and Medicines for Malaria Venture (MMV08/0105)
Keywords:
Fungal
metabolites,
Naegleria
fowleri
and
Acanthamoeba
spp,
Dirofilaria
immitis,
Brugia
panangi,
Brugia
malayi,
Plasmodium
falciparum,
Strongyloides
stercoralis,
Haemonchus
contortus
P627
A
UHPLC/MS-‐MS-‐based
HDAC
assay
applied
to
bio-‐guided
micro-‐
fractionation
of
fungi
extracts
Vincent
Zwick,
Claudia
A.
Simões-‐Pires,
Lucie
Ory,
Pierre-‐Marie
Allard,
Jean-‐Luc
Wolfender,
Muriel
Cuendet
School
of
Pharmaceutical
Sciences,
University
of
Geneva,
University
of
Lausanne,
1211
Geneva,
Switzer-‐
land
Histone
acetylation
and
deacetylation
constitutes
a
dynamic
and
reversible
process
catalyzed
by
two
classes
of
enzymes,
histone
acetyltransferase
(HATs)
and
histone
deacetylases
(HDACs),
respectively.
To
date,
four
HDAC
pan-‐inhibitors,
vorinostat,
romidepsin,
belinostat,
and
panobinostat,
have
been
approved
by
the
FDA
for
cancer
treatment
[1].
Natural
products
have
been
of
great
contribution
to
the
early
discovery
of
potent
HDAC
inhibitors.
Romidepsin,
for
example,
has
been
originally
isolated
from
Chromobacterium
violaceum
[2].
However,
the
discovery
of
natural
HDAC
inhibitors
has
not
been
driven
by
HDAC
bioguided
fractionation
so
far.
Instead,
HDAC
inhibitory
activity
of
many
natural
compounds
has
been
discovered
in
a
second
step
following
anti-‐tumor
activity
screening
[2,
3].
Indeed,
the
most
commonly
used
fluorescence-‐based
HDAC
activity
assays
are
not
well
suited
for
screening
crude
extracts
and
fractions
obtained
from
nature.
To
accelerate
the
search
for
HDAC
inhibitors
of
natural
origin,
a
UHPLC-‐MS/MS-‐based
enzymatic
assay
was
optimized
and
applied
to
the
bioactivity-‐guided
identification
of
new
HDAC
inhibitors
in
fungi.
By
this
means,
salirepol
was
isolated
and
iden-‐
tified
as
a
new
HDAC
inhibitor
from
Penicillium
griseofulvum.
Moreover,
salirepol
showed
se-‐
lective
HDAC6
inhibition
in
the
low
micromolar
range
(IC50
=
3.4
µM).
The
UHPLC-‐MS/MS
approach
was
compared
to
the
traditional
fluorescence-‐based
assay,
which
led
to
highlighting
the
advantages
and
the
applicability
of
the
MS-‐based
method.
Acknowledgements:
Laurence
Marcourt
is
acknowledged
for
NMR
experiments
Keywords: UHPLC/MS-‐MS, HDAC inhibitor, fungi, salirepol
References:
[1] Mottamal
M,
Zheng
S,
Huang
T,
Wang
G.
Histone
deacetylase
inhibitors
in
clinical
studies
as
templates
for
new
anticancer
agents.
Molecules
2015;
20:
3898-‐3941
[2] Ueda
H,
Manda
T,
Matsumoto
S,
Mukumoto
S,
Nishigaki
F,
Kawamura
I,
Shimomura
K.
FR901228,
a
novel
antitumor
bicyclic
depsipeptide
produced
by
Chromobacterium
vio-‐
laceum
III.
Antitumor
activities
on
experimental
tumors
in
mice.
J
Antibiot
(Tokyo)
1994;
47:
315-‐323
[3] Nakajima
H,
Kim
YB,
Terano
H,
Yoshida
M,
Horinouchi
S.
FR901228,
a
potent
antitumor
antibiotic,
is
a
novel
histone
deacetylase
inhibitor.
Exp
Cell
Res
1998;
241:
126-‐133
P628
A
cytotoxic
pentadecapeptide
from
a
South
African
Didemnid
tu-‐
nicate
David
Gallegos1,
Jeffrey
Serrill1,
Shirley
Parker-‐Nance2,
Rosemary
Dorrington3,
Jane
Ishmael1,
Kerry
McPhail1
1
Department
of
Pharmaceutical
Sciences,
College
of
Pharmacy,
Oregon
State
University,
Corvallis,
OR
97331,
USA,
2
South
African
Institute
of
Aquatic
Biosciences
and
Nelson
Mandela
Metropolitan
University,
3
Rhodes
University,
Eastern
Cape,
South
Africa
The
rate
of
discovery
of
new
natural
product
chemical
entities
has
plateaued,
and
unique
populations
of
endemic,
biologically
diverse
sessile
marine
organisms
represent
increasingly
critical
opportunities
to
discover
new
chemistry.
Discovery
of
the
mandelalides
[1]
as
potent
inhibitors
of
cancer
cell
growth
from
the
new
South
African
tunicate
Lissoclinum
mandelai
is
an
example
of
the
diverse
suites
of
metabolites
with
potent
biological
activities
that
have
been
isolated
from
tunicates
and
other
filter-‐feeding
sessile
marine
organisms
that
house
complex
microbial
consortia.
Further
investigation
of
archived
and
new
tunicate
collections
from
Algoa
Bay,
South
Africa,
has
revealed
a
group
of
didemnid
tunicates
with
an
unusual
gelatinous
morphology
similar
to
Lissoclinum
mandelai.
Using
a
bioassay-‐guided
isolation
approach,
a
new
“gelatinous”
species
of
the
genus
Didemnum
has
yielded
a
cytotoxic
pentadecapeptide
with
a
molecular
mass
of
1603.7688
Da,
comprising
fifteen
residues
including
both
proteino-‐
genic
and
non-‐proteinogenic
amino
acids.
The
pure
compound
inhibited
both
HeLa
cervical
cancer
and
NCI-‐H460
non-‐small
cell
lung
cancer
cell
lines
when
tested
at
30
nM
in
prelimi-‐
nary
assays
against
cells
seeded
at
low
densities.
Inhibition
of
cancer
cells
at
low
starting
den-‐
sity
may
be
indicative
of
an
anti-‐proliferative
mechanism
of
action.
The
compound
did
not
show
antibacterial
activity
against
Vibrio
cholera.
Didemnin
B
and
its
clinically
approved
ana-‐
logue
dehydrodidemnin
B
(plitidepsin,
Aplidin®)
[2,
3]
are
important
macrocyclic
depsipep-‐
tides
from
a
didemnid
tunicate.
The
pentadecapeptide
reported
here
provides
justification
for
our
continued
investigation
of
unique,
endemic
didemnid
tunicates
from
South
Africa
as
a
source
of
new
macrocyclic
natural
products
with
cytotoxic,
anti-‐viral
or
antimicrobial
activity.
Acknowledgements:
We
acknowledge
the
South
African
government
for
permission
to
collect
the
subject
tunicate
(Collection
Permit
No.
278
RES2013/43)
Keywords: Tunicates, natural products, cytotoxic, peptide, macrocycle, didemnid
References:
[1]
Sikorska
J,
Hau
AM,
Anklin,
C,
Parker-‐Nance
S,
Davies-‐Coleman
MT,
Ishmael
JE,
McPhail
KL,
Mandelalides
AD.
Cytotoxic
macrolides
from
a
new
Lissoclinum
species
of
South
African
tunicate.
J
Org
Chem
2012;
77:
6066-‐6075
[2]
Shin
DM,
Holoye
PY,
Forman
A,
Winn
R,
Perez-‐Soler
R,
Dakhil
S,
Rosenthal
J,
Raber
MN,
Hong
WK.
Phase
II
clinical
trial
of
didemnin
B
in
previously
treated
small
cell
lung
cancer.
Invest
New
Drugs
1994;
12:
243-‐249
[3]
Urdiales
J,
Morata
P,
Castro
IND,
Sánchez-‐Jiménez
F.
Antiproliferative
effect
of
dehydro-‐
didemnin
B
(DDB),
a
depsipeptide
isolated
from
Mediterranean
tunicates.
Cancer
Lett
1996;
102:
31-‐37
P629
Insect
symbionts:
Prolific
sources
of
new
bioactive
compounds
Dong-‐Chan
Oh
Natural
Products
Research
Institute,
College
of
Pharmacy,
Seoul
National
University,
1
Gwanak-‐ro,
Gwa-‐
nak-‐gu,
Seoul
08826,
Republic
of
Korea
Symbioses
between
insects
and
microorganisms
are
widespread
in
nature.
Recent
studies
of
the
secondary
metabolites
of
insect
symbionts
have
revealed
that
insect
symbionts
could
be
tremendous
sources
of
new
bioactive
compounds
with
pharmaceutical
potential
[1].
In
this
presentation,
the
discovery
of
new
secondary
metabolites
from
symbionts
in
diverse
insects
such
as
the
dung
beetle
(Copris
tripartitus),
the
silkworm
(Bombyx
mori),
and
the
burying
bee-‐
tle
(Nicrophorus
concolor),
the
horn
beetle
(Allomyrina
dichotoma),
and
the
carpenter
(Com-‐
ponotus
japonicus)
ant
will
be
discussed.
For
example,
the
coprisamides
A
and
B,
isolated
from
a
Streptomyces
sp.
strain
from
the
gut
of
the
dung
beetle,
turned
out
to
be
structurally
unique
cyclic
peptides
bearing
a
couple
of
modi-‐
fied
amino
acids
(β-‐methyl-‐aspartic
acid
and
diaminopropanoic
acid),
three
D-‐amino
acids,
and
an
unusual
2-‐alkenyl
cinnamic
acid.
The
branched
feature
of
the
coprisamides
at
dia-‐
minopropanoic
acid
to
valine
is
also
biosynthetically
interesting
[2].
In
addition,
diverse
new
bioactive
compounds
such
as
antibacterial
macrocyclic
lactams
and
unusual
cyclic
hexapep-‐
tides
were
discovered
from
the
symbiotic
bacteria
in
the
silkworm
and
the
burying
beetle,
respectively.
The
discovery
of
structurally,
biologically,
and
biosynthetically
interesting
sec-‐
ondary
metabolites
from
insect
symbionts
demonstrates
that
studying
insect
symbionts
in
search
for
new
bioactive
compounds
could
be
a
new
promising
strategy
in
natural
product
research.
Coprisamide A
Acknowledgements:
This
work
was
supported
by
National
Research
Foundation
of
Korea
(NRF)
grants
funded
by
the
Korean
government
(Ministry
of
ICT
and
Future
Planning)
(2014R1A2A1A11053477
and
2009-‐0083533)
and
HHMI
International
Early
Career
Scientist
Program.
Keywords: Insect, symbiont, secondary metabolite, antibacterial
References:
[1] Bode
HB.
Insects:
True
pioneers
in
anti-‐infective
therapy
and
what
we
can
learn
from
them.
Angew
Chem
Int
Ed
2009;
48:
6394-‐6396
[2] Um
S,
Park
SH,
Kim
J,
Park
HJ,
Ko
K,
Bang
HS,
Lee
SK,
Shin
J,
Oh
D-‐C.
Coprisamides
A
and
B,
new
cyclic
peptides
from
a
gut
bacterium
of
the
dung
beetle
Copris
tripartitus.
Org
Lett
2015;
17:
1272-‐1275
P630
Characterisation
of
vietnamycin:
a
novel
Burkholderia
antibiotic
targeting
mupirocin-‐resistant
methicillin-‐resistant
Staphylococ-‐
cus
aureus
(MRSA)
Rachel
A.
Rowe1,
Cerith
Jones1,
Matthew
J.
Bull1,
Matthew
Jenner2,
Lijang
Song2,
Yousef
Dash-‐
ti2,
Simon
R.
Harris3,
Julian
Parkhill3,
Thomas
R.
Connor1,
Gregory
L.
Challis2,
Eshwar
Ma-‐
henthiralingam1
1
School
of
Biosciences,
Cardiff
University,
Cardiff,
Wales,
UK,
2
School
of
Chemistry,
University
of
War-‐
wick,
UK,
3
Wellcome
Trust
Sanger
Institute,
Wellcome
Trust
Genome
Campus,
Hinxton,
Cambridge,
UK
Gram-‐negative
Burkholderia
bacteria
are
an
untapped
source
of
new
antibiotics
with
multiple
bioactive
compounds
currently
under
investigation1.
An
antimicrobial
activity
produced
by
Burkholderia
vietnamiensis
has
been
identified
and
designated
vietnamycin.
The
compound
has
potent
activity
against
methicillin-‐resistant,
mupirocin-‐resistant
Staphylococcus
aureus
(MRSA),
but
the
chemical
structure
of
vietnamycin
and
its
biosynthetic
pathway
are
not
known.
To
bring
vietnamycin
from
discovery
towards
pre-‐clinical
testing,
an
efficient
extrac-‐
tion
method
based
on
solvent
extraction
of
growth
media
followed
by
FLASH
chromatography
has
been
developed.
A
thin
layer
chromatography
(TLC)
bioassay
of
the
resulting
crude
ex-‐
tract
demonstrated
the
presence
of
two
bioactive
fractions,
with
vietnamycin
(Rf
0.43)
sepa-‐
rating
from
a
bioactive
fraction
with
a
mobility
(Rf
0.90)
characteristic
of
the
4-‐hydroxy-‐3-‐
methyl-‐2-‐alkylquinoline
(HMAQ)
Burkholderia
signalling
molecules1.
In
tandem
with
chemi-‐
cal
characterisation
of
vietnamycin,
a
genome
mining
approach
is
being
undertaken
to
identi-‐
fy
the
gene
cluster
responsible
for
its
biosynthesis.
The
genome
sequence
of
producer
strain
B.
vietnamiensis
BCC0268
(6.77
Mb)
has
been
determined
by
single
molecule
real
time
se-‐
quencing,
and
is
composed
of
3
chromosomal
replicons
(3.28,
2.22,
and
1.26
Mb).
Prediction
of
secondary
metabolite
biosynthesis
clusters
was
performed
using
the
Antibiotics
and
Sec-‐
ondary
Metabolite
Analysis
Shell2
(antiSMASH),
and
demonstrated
presence
of
32
clusters
including
a
nonribosomal
peptide
synthase
(NRPS)
and
NRPS-‐type
I
polyketide
synthase
(PKS)
locus.
Deletion
of
the
smallest
replicon
had
no
impact
on
vietnamycin
biosynthesis,
eliminating
the
NRPS-‐type
I
PKS
encoded
on
this
replicon
as
the
pathway
responsible
for
its
biosynthesis.
In
summary,
an
interdisciplinary
approach
combining
microbiology,
chemistry,
and
genome
mining
is
being
used
to
identify
vietnamycin
and
progress
it
as
a
novel
Burkhold-‐
eria
antibiotic.
Acknowledgements:
This
work
was
funded
by
Biotechnology
and
Biology
Research
Council
(grant
BB/L021692/1)
and
R.
Rowe
is
the
recipient
of
a
PhD
studentship
from
the
Life
Sciences
Research
Net-‐
work
Wales
Keywords: Antibiotics, Burkholderia vietnamiensis, MRSA, genome mining
References:
[1]
Mahenthiralingam
E,
Song
L,
Sass
A,
White
J,
Wilmot
C,
Marchbank
A,
Boaisha
O,
Paine
J,
Knight
D,
Challis
GL.
Enacyloxins
are
products
of
an
unusual
hybrid
modular
polyketide
synthase
encoded
by
a
cryptic
Burkholderia
ambifaria
Genomic
Island.
Chem
Biol
2011;
18:
665-‐677
[2]
Blin
K,
Medema
MH,
Kazempour
D,
Fischbach
MA,
Breitling
R,
Takano
E,
Weber
T.
an-‐
tiSMASH
2.0
-‐
a
versatile
platform
for
genome
mining
of
secondary
metabolite
producers.
Nucleic
Acids
Res
2013;
41:
204-‐212
P631
New
antiplasmodial
compounds
discovered
by
High
Throughput
Screening
(HTS)
of
a
collection
of
microbial
natural
extracts
Noureddine
El
Aouad1,
Frederick
Annang1,
Ignacio
Pérez-‐Victoria1,
Jesús
Martín1,
Gloria
Cre-‐
spo1,
Elizabeth
Domingo1,
Guiomar
Pérez-‐Moreno2,
Juan
Cantizani1,
Paula
Sánchez-‐Carrasco2,
Ignacio
González1,
Víctor
González-‐Menéndez1,
Nuria
de
Pedro1,
José
R.
Tormo1,
Luis
M.
Ruiz-‐
Pérez2,
Dolores
González-‐Pacanowska2,
Francisca
Vicente1,
Gerald
F.
Bills1,
Olga
Genilloud1,
Fernando
Reyes1
1
Fundación
MEDINA,
Avenida
del
Conocimiento
34,
18016-‐Armilla,
Granada,
Spain,
2
Instituto
de
Para-‐
sitología y Biomedicina “López-‐Neyra”. CSIC. Avda del Conocimiento s/n, 18016-‐Armilla, Granada, Spain
Malaria
is
a
widespread
disease
in
tropical
and
subtropical
regions
and
an
estimated
3.2
bil-‐
lion
people
are
at
the
risk
of
suffering
it,
with
0.5-‐1
million
deaths
reported
in
2014.
Due
to
the
low
variety
and
structural
diversity
of
antimalarial
drugs
currently
available
in
the
clinic
and
the
increasing
number
of
cases
of
resistance
against
these
drugs,
there
is
an
urgent
need
to
find
new
treatments
with
novel
modes
of
action.
Microbial
natural
products
have
not
been
yet
exhaustively
explored
in
the
search
for
new
antiparasitic
drugs,
especially
in
the
case
of
malaria,
and
therefore
they
offer
a
valuable
source
for
the
discovery
of
new
and
interesting
hits
[1].
With
the
aim
of
finding
new
drugable
natural
products
with
antimalarial
properties,
a
subset
of
22,000
samples
of
the
MEDINA
natural
product
extracts
collection,
one
of
the
largest
natural
products
libraries
harbouring
more
than
130,000
microbial
extracts,
was
screened
using
a
phenotypic
HTS
based
on
measurements
of
Plasmodium
falciparum
lactate
dehydro-‐
genase
[2].
Active
hits
from
this
test
were
subjected
to
chemical
dereplication
using
LC-‐LRMS,
LC-‐HRMS
and
LC-‐NMR
in
the
off-‐line
mode
in
order
to
discard
samples
containing
known
an-‐
timalarial
and/or
cytotoxic
molecules
[3].
Hits
containing
potentially
new
molecules
were
re-‐
fermented
at
1L
scale
and
the
compounds
responsible
for
the
bioactivity
of
the
extracts
were
isolated
following
a
bioassay-‐guided
process.
Several
new
molecules
with
antimalarial
activity
in
the
micromolar
range
were
obtained
using
this
approach,
including
the
naphthoquinone
lasionectrin
[4],
the
betaine
lipid
MDN-‐0104
[5],
a
new
member
of
the
pepstatin
family,
pep-‐
statin
K
[2],
and
two
cyclic
hexapeptides.
Additionally,
a
new
polycyclic
xanthone
structurally
related
to
xantholipin
[6]
and
Sch
54445
[7]
with
an
IC50
of
9
nM
against
P.
falciparum
3D7
was
also
obtained.
The
approach
used
in
this
work
confirms
once
more
that
microbial
natural
products
continue
to
be
a
rich
and
underexploited
source
of
novel
molecules
that
might
lead
to
the
discovery
of
new
and
interesting
drugs.
Acknowledgements:
This
work
was
supported
by
the
Junta
de
Andalucía
[BIO-‐199,
P09-‐CVI-‐
5367],
the
VI
Plan
Nacional
de
Investigación
Científica,
Desarrollo
e
Innovación
Tecnológica
2008-‐2011,
Instituto
de
Salud
Carlos
III-‐Subdirección
General
de
Redes
y
Centros
de
Investigación
Cooperativa-‐Red
de
Investi-‐
gación
Cooperativa
en
Enfermedades
Tropicales
(RICET
FIS
Network:
RD12/0018/0017),
the
Plan
Nacional
(SAF2013-‐48999-‐R),
the
FEDER
funds
from
the
EU
and
the
PARAMET
network
(FP7-‐PEOPLE-‐
2011-‐ITN.
GA290080)
Keywords: Malaria, Plasmodium falciparum, HTS, microbial extracts, natural products
References:
[1] Kaur
K,
Jain
M,
Kaur
T,
Jain
R.
Antimalarials
from
nature.
Bioorg
Med
Chem
2009;
17:
3229–3256.
[2] Pérez-‐Moreno
G,
Cantizani
J,
Sánchez-‐Carrasco
P,
Ruiz-‐Pérez
LM,
Martín
J,
El
Aouad
N,
Pérez-‐Victoria
I,
Tormo
JR,
González
V,
González
I,
de
Pedro
N,
Reyes
F,
Genilloud
O,
Vi-‐
cente
F,
González-‐Pacanowska
D.
Discovery
of
new
compounds
active
against
Plasmodi-‐
um
falciparum
by
high
throughput
screening
of
microbial
natural
products.
PLoS
ONE
2016;
11:
e0145812
[3] Pérez-‐Victoria
I,
Martín
J,
Reyes
F.
Combined
LC/UV/MS
and
NMR
strategies
for
the
derep-‐
lication
of
marine
natural
products.
Planta
Med,
advance
online
publication
22
March
2016;
doi:
10.1055/s-‐0042-‐101763
[4] El
Aouad
N,
Pérez-‐Moreno
G,
Sánchez
P,
Cantizani
J,
Ortiz-‐López
FJ,
Martín
J,
González-‐
Menéndez
V,
Ruiz-‐Pérez
LM,
González-‐Pacanowska
D,
Vicente
F,
Bills
G,
Reyes
F.
La-‐
sionectrin,
a
Naphthopyrone
from
a
Lasionectria
sp.
J
Nat
Prod
2012;
75:
1228-‐1230.
[5] Martín
J,
Crespo
G,
González-‐Menéndez
V,
Pérez-‐Moreno
G,
Sánchez-‐Carrasco
P,
Pérez-‐
Victoria
I,
Ruiz-‐Pérez
LM,
González-‐Pacanowska
D,
Vicente
F,
Genilloud
O,
Bills
GF,
Reyes
F.
MDN-‐0104,
an
antiplasmodial
betaine
lipid
from
Heterospora
chenopodii.
J
Nat
Prod
2014;
77:
2118-‐2123.
[6] Terui
Y,
Yiwe,
C,
Jun-‐ying
L,
Ando
T,
Yamamoto
H,
Kawamura
Y,
Tomishima
Y,
Uchida
S,
Okazaki,
T,
Munetomo
E,
Seki
T,
Yamamoto
K,
Murakami
S,
Kawashima
A.
Xantholipin,
a
novel
inhibitor
of
HSP47
gene
expression
produced
by
Streptomyces
sp.
Tetrahedron
Lett
2003;
44:
5427-‐5430.
[7] Chu
M,
Truumees
I,
Mierzwa
R,
Terracciano
J,
Patel
M,
Loebenberg
D,
Kaminski
JJ,
Das
P,
Puar
MS.
Sch
54445:
A
new
polycyclic
xanthone
with
highly
potent
antifungal
activity
produced
by
Actinoplanes
sp.
J
Nat
Prod
1997;
60:
525-‐528.
P632
Decalin-‐containing
polyketides
with
antibacterial
activities
from
an
endophytic
fungus,
Eupenicillium
sp.
LG41
Gang
Li1,
Souvik
Kusari1,
Hartmut
Laatsch2,
Christopher
Golz3,
Carsten
Strohmann3,
Michael
Spiteller1
1
Institute
of
Environmental
Research
(INFU),
Department
of
Chemistry
and
Chemical
Biology,
Chair
of
Environmental
Chemistry
and
Analytical
Chemistry,
TU
Dortmund,
Otto-‐Hahn-‐Straße
6,
44221
Dort-‐
mund,
Germany,
2
Institute
for
Organic
and
Biomolecular
Chemistry,
Georg-‐August
University,
Tam-‐
mannstraße
2,
37077
Göttingen,
Germany,
3
Inorganic
Chemistry,
Department
of
Chemistry
and
Chemical
Biology,
TU
Dortmund,
Otto-‐Hahn-‐Straße
6,
44221
Dortmund,
Germany
Endophytes
have
been
well-‐known
to
synthesize
diverse
and
novel
secondary
metabolites
with
intriguing
biological
activities
[1,
2].
LC-‐HRMS
guided
chemical
research
on
an
endophyt-‐
ic
fungus,
Eupenicillium
sp.
LG41
isolated
from
traditional
medicinal
plant
Xanthium
sibiricum,
afforded
one
known
(1)
and
four
new
(2-‐5)
decalin
moiety-‐containing
polyketides
[3,
4].
The
structures
and
stereochemistry
of
the
new
compounds
were
elucidated
by
extensive
spectro-‐
scopic
analysis
using
LC-‐HRMS,
NMR,
optical
rotation,
and
ECD
calculation,
as
well
as
single
crystal
X-‐ray
diffraction.
Compounds
4
and
5
existed
in
chemical
equilibrium
in
two
and
three
cis/trans
isomers,
respectively,
as
indicated
by
LC-‐HRMS
and
NMR.
Compound
3
exhibited
antibacterial
activity
against
the
clinically
relevant
Staphylococcus
au-‐
reus
with
an
MIC
value
of
1.0
µg/mL.
Compound
5
not
only
was
highly
active
against
S.
aureus
with
an
MIC
value
of
0.1
µg/mL,
but
also
demonstrated
marked
cytotoxicity
against
human
acute
monocytic
leukemia
cell
line
(THP-‐1).
These
data
reveal
that
altering
the
substitution
at
C-‐11
could
drastically
increase
the
antibacterial
activity.
Acknowledgements:
The
Ministry
of
Innovation,
Science,
Research
and
Technology
of
the
State
of
North
Rhine-‐Westphalia,
Germany,
and
the
German
Research
Foundation
(DFG)
are
thankfully
acknowledged
for
granting
a
high-‐resolution
mass
spectrometer.
G.L.
gratefully
acknowledges
the
China
Scholarship
Council
(CSC)
for
a
doctoral
fellowship
Keywords: Endophytes, secondary metabolites, polyketides, decalin, antibacterial activity
References:
[1] Kusari
S,
Hertweck
C,
Spiteller
M.
Chemical
ecology
of
endophytic
fungi:
origins
of
second-‐
ary
metabolites.
Chem
Biol
2012;
19:
792-‐798
[2] Li
G,
Kusari
S,
Kusari
P,
Kayser
O,
Spiteller
M.
Endophytic
Diaporthe
sp.
LG23
produces
a
potent
antibacterial
tetracyclic
triterpenoid.
J
Nat
Prod
2015;
78:
2128-‐2132
[3] Li
G,
Kusari
S,
Spiteller
M.
Natural
products
containing
‘decalin’
motif
in
microorganisms.
Nat
Prod
Rep
2014;
31:
1175-‐1201
[4] Li
G,
Kusari
S,
Lamshöft
M,
Schüffler
A,
Laatsch
H,
Spiteller
M.
Antibacterial
secondary
me-‐
tabolites
from
an
endophytic
fungus,
Eupenicillium
sp.
LG41.
J
Nat
Prod
2014;
77:
2335-‐
2341
P632a
An
endophytic
fungus
Phyllosticta
capitalensis
harboring
unculti-‐
vable
endosymbiotic
bacterium
Herbaspirillum
sp.
produces
lac-‐
tam-‐fused
4-‐pyrones
Wen-‐Xuan
Wang1,
Souvik
Kusari1,
Parijat
Kusari2,
Oliver
Kayser2,
Michael
Spiteller1
1
Department
of
Chemistry
and
Chemical
Biology,
Chair
of
Environmental
Chemistry
and
Analytical
Chemistry,
Institute
of
Environmental
Research
(INFU),
TU
Dortmund,
Otto-‐Hahn-‐Straße
6,
44221
Dort-‐
mund,
Germany,
2
Department
of
Biochemical
and
Chemical
Engineering,
Chair
of
Technical
Biochemis-‐
try,
TU
Dortmund,
Emil-‐Figge-‐Straße
66,
44227
Dortmund,
Germany
Unraveling
the
chemical
and
molecular
interactions
occurring
between
endophytic
microor-‐
ganisms
in
their
natural
niches
is
an
essential
prerequisite
for
exploiting
their
potential
as
sources
of
untapped
natural
products
[1,
2].
Herein
we
report
the
isolation
and
characteriza-‐
tion
of
two
new
lactam-‐fused
4-‐pyrones
by
an
endophytic
fungus
Phyllosticta
capitalensis,
which
harbors
an
uncultivable
endosymbiotic
bacterium
Herbaspirillum
sp.,
isolated
from
the
boxwood
plant
Buxus
sinica
[Buxaceae].
Their
structures
were
elucidated
by
LC-‐HRMSn,
1D
and
2D
NMR,
DFT
13C
NMR
calculation
and
chemical
reaction.
On
the
basis
of
their
structures,
they
should
comprise
of
a
combination
of
polyketide
synthase
and
non-‐ribosomal
peptide
synthetase,
leading
to
the
PKS-‐NRPS
hybrids.
For
the
identification
of
PKS
and
NRPS
genes
in
the
fungal
metagenome,
a
degenerate
primer-‐based
approach
was
first
used
based
on
al-‐
ready-‐reported
fungal
and
bacterial
genes
encoding
the
highly
conserved
ketosynthase
do-‐
main
(KS)
and
the
conserved
adenylation
domain
(A),
respectively.
This
revealed
that
the
compounds
were
plausibly
biosynthesized
using
the
fungal
PKS
and
the
bacterial
NRPS.
We
are
presently
in
the
process
of
sequencing
the
entire
metagenome
of
the
endophytic
fungus
containing
the
endosymbiont,
which
will
confirm
the
molecular
pathway
of
biogenesis
of
the-‐
se
two
compounds.
The
interface
of
plants
and
endophytic
communities
are
complex
ecologi-‐
cal
systems.
The
scenario
of
endophytes
containing
uncultivable
endosymbionts,
further
add
to
a
higher
level
of
complexity
operational
in
natural
ecological
niches,
which
could
be
anoth-‐
er
challenge
for
the
future
investigations.
These
hidden
endosymbiotic
endophytes
and
their
corresponding
functions
are
likely
essential
parts
of
the
whole
interaction
network
at
the
plant-‐microbe
interface.
Acknowledgements:
We
have
to
thank
the
Ministry
of
Innovation,
Science,
Research
and
Technology
of
the
State
of
North
Rhine-‐Westphalia,
Germany
and
the
German
Research
Foundation
(DFG)
for
funding
a
high-‐resolution
mass
spectrometer.
W-‐WW
is
grateful
to
the
China
Scholarship
Council
(CSC)
for
a
doc-‐
toral
fellowship.
We
gratefully
acknowledge
Dr.
W.
Hiller
(Department
of
Chemistry
and
Chemical
Biolo-‐
gy,
TU
Dortmund)
for
the
realization
of
the
NMR
measurements,
and
Dr.
S.
Zühlke
(INFU,
TU
Dortmund)
for
valuable
discussions
and
realization
of
mass
spectrometric
analyses
Keywords: Endophytes, Buxus sinica, Phyllosticta capitalensis, Herbaspirillum sp., endosym-‐
biont
References:
[1] Kusari
S,
Hertweck
C,
Spiteller
M.
Chemical
ecology
of
endophytic
fungi:
origins
of
second-‐
ary
metabolites.
Chem
Biol
2012;
19:
792−798
[2] Kusari
S,
Spiteller,
M.
Metabolomics
of
endophytic
fungi
producing
associated
plant
sec-‐
ondary
metabolites:
progress,
challenges
and
opportunities.
In:
Roessner
U,
editor.
Metabolomics.
Rijeka,
Croatia:
InTech
2012;
241−266
P633
Two
new
compounds
produced
by
Xylaria
sp.
an
endophytic
fun-‐
gus
from
Casearia
sylvestris
(Flacourtiaceae)
G.
F.
Martins1,
C.
R.
Nogueira1,
G.
H.
Silva1,
M.
C.
M.
Young2,
C.
O.
Pessoa3,
D.
J.
B.
Lima3,
P.
M.
P.
Ferreira3,
V.
S.
Bolzani1,
A.
R.
Araujo1
1
NuBBE
–
Nucleus
of
Bioassays,
Biosynthesis
and
Ecophysiology
of
Natural
Products,
State
University
of
São
Paulo,
Institute
of
Chemistry,
Department
of
Organic
Chemistry,
Zip
Code
14800-‐900,
Araraquara,
São
Paulo,
Brazil,
2
Section
of
Physiology
and
Biochemistry
of
plants,
Botany
Institute,
Zip
Code
01061-‐
970,
São
Paulo,
Brazil,
3
Experimental
Oncology
Laboratory,
Department
of
Biophysics
and
Physiology,
Federal
University
of
Piauí,
Zip
Code
64049-‐550,
Teresina,
Piauí,
Brazil.
Endophytic
fungi
are
widespread
in
nature
and
attract
considerable
attention
as
prolific
sources
of
new
natural
products
with
diverse
biological
properties
and
structures,
including
antibacterial,
antifungal,
antiinflammatory,
and
antitumor
activities
[1,
2].
In
particular,
fungi
of
the
genus
Xylaria
sp.
belonging
to
Xylariaceae
family
(Ascomicota),
have
shown
to
be
po-‐
tential
sources
of
new
secondary
metabolites,
most
of
them
with
biological
activities
[3].
In
our
research
for
bioactive
compounds
from
endophytic
fungi
of
Brazilian
plants,
Xylaria
sp.
has
been
isolated
from
Casearia
sylvestris
(Flacourtiaceae).
Xylaria
sp.
was
cultured
in
rice
for
21
days
at
26
oC
under
static
mode.
The
culture
was
extracted
with
EtOAc.
After
evaporation
of
the
solvent,
the
crude
EtOAc
extract
was
partioned
using
water
by
liquid
partitioning.
The
EtOAc
fraction
was
evaporated
resulting
in
crude
extract.
This
extract
was
dissolved
in
CH3CN
and
defatted
with
hexane
by
liquid
partitioning.
Following,
the
CH3CN
fraction
was
evapo-‐
rated
to
give
0.249
g
of
the
crude
extract.
The
crude
extract
exhibited
activity
against
three
cancer
cell
lines
(ovarian
carcinoma,
colon
and
glioblastoma),
strong
activity
against
fungal
pathogens
C.
cladosporioides
and
C.
sphaerospermum,
and
moderate
inhibitory
activity
of
ace-‐
tylcholinesterase
enzyme.
The
CH3CN
extract
was
subjected
to
chromatographic
separation
by
preparative
HPLC
(reversed-‐phase)
yielding
the
isolation
of
two
new
compounds,
an
α-‐
pyrone
(5-‐butyl-‐6-‐hydroxy-‐methyl-‐2-‐pyrone)
and
an
isocoumarin
(6-‐hydroxy-‐7,8-‐dimethoxy-‐
3-‐methyl-‐3,4-‐dihiydro-‐isocoumarin.
The
structure
elucidation
of
these
compounds
was
achieved
by
spectroscopic
data,
including
HREIMS,
1D
and
2D
NMR
(HSQC,
HMBC,
COSY,
NO-‐
ESY).
The
isolated
compounds
are
under
biological
investigation.
Acknowledgements:
CNPq,
FAPESP,
and
CAPES
Keywords: Endophytic fungi, Xylaria sp., biological activities, pyrone, isocoumarin
References:
[1] Aly
AH,
Debbab
A,
Proksch
P.
Fungal
endophytes:
unique
plant
inhabitants
with
great
promises.
Appl
Microbiol
Biotechnol
2011;
90:
1829-‐1845
[2] Jalgaonwal
RE,
Mohite
BV,
Mahajan
RT.
A
review:
Natural
products
from
plant
associated
endophytic
fungi.
J
Microbiol
Biotechnol
Res
2011
;
1:
21-‐32
[3] Song,
F,
Wu
SH,
Zhai
YZ,
Xuan
QC,
Wang
T.
Secondary
metabolites
from
the
genus
Xylaria
and
their
bioactivities.
Chem
Biodivers
2014;
11:
673-‐699
P634
Antifungal
long-‐chain
alkenyl
sulphates
isolated
from
culture
broths
of
the
fungus
Chaetopsina
sp.
Gloria
Crespo,
Ignacio
Pérez
Victoria,
Mercedes
de
la
Cruz,
Víctor
González-‐Menéndez,
Bastien
Cautain,
Jesús
Martín,
Francisca
Vicente,
Olga
Genilloud,
Fernando
Reyes
Fundación
MEDINA,
Avda.
del
Conocimiento
34,
Parque
Tecnológico
de
Ciencias
de
la
Salud,
E-‐18016,
Armilla,
Granada,
Spain
Invasive
infections
caused
by
fungi
have
experienced
a
great
increase
in
recent
years,
mainly
due
to
the
rising
number
of
immunocompromised
patients.
The
therapeutic
arsenal
of
anti-‐
fungal
drugs
currently
in
use
is
rather
limited
and
the
evolving
threat
of
resistant
emerging
pathogens
has
turned
the
development
of
new
antifungal
agents,
preferably
naturally-‐
occurring
compounds
with
novel
mechanisms
of
action,
low
resistance
rates
and
fewer
side
effects,
into
an
urgent
medical
need.
As
part
of
our
program
focused
on
the
discovery
and
characterization
of
new
antifungal
com-‐
pounds,
a
total
of
8320
extracts
from
our
collection
were
tested
against
the
fungal
parasites
Candida
glabrata,
C.
krusei,
C.
parapsilosis,
C.
tropicalis,
C.
albicans
and
Aspergillus
fumigatus.
A
total
of
127
extracts
displayed
antifungal
properties
and
after
LC/MS
dereplication,
10
were
selected
for
further
fractionation.
One
of
them,
the
acetone
extract
of
fungus
Chaetopsina
sp.,
isolated
from
leaf
litter
of
the
plant
Beilschmiedia
tawa
(from
Basil
Hewett
Reserve),
grown
in
STP
medium
displayed
antifungal
properties.
Bioassay-‐guided
fractionation
from
a
1L
fer-‐
mentation,
using
a
combination
of
low
resolution
chromatography
on
SP207ss
resin
and
re-‐
versed
phase
HPLC,
led
to
the
isolation
of
linoleyl
sulphate
and
two
structurally
related
con-‐
geners
as
the
compounds
responsible
for
the
observed
activity.
Linoleyl
sulphate
has
been
isolated
for
the
first
time
as
a
natural
product.
This
molecule
was
previously
described
as
a
synthetic
product
with
the
ability
to
produce
the
allosteric
inhibition
of
soybean
(SLO)
and
human
(15-‐HLO)
lipoxygenase,
a
potential
target
for
therapies
against
cancer,
asthma
and
artherosceloris
[1].
The
natural
sulphates
isolated
displayed
antifungal
activity
against
the
panel
of
six
fungal
species
cited
above,
with
MIC
values
ranging
between
2
and
64
µg/mL.
In
conclusion
a
group
of
antifungal
sulphated
long
chain
alcohols
has
been
isolated
from
cultures
of
Chaetopsina
sp.
To
the
best
of
our
knowledge,
this
is
the
first
time
that
compounds
belonging
to
this
structural
class
have
been
isolated
from
microbial
sources.
Keywords: Long-‐chain sulphates, Chaetopsina sp., antifungal, structural elucidation.
References:
[1] Mogul
R,
Holman
T
R.
Inhibition
studies
of
soybean
and
human
15-‐lipoxygenases
with
long-‐chain
alkenyl
sulfated
substrates.
Biochemistry
2001;
40:
4391−4397
P635
Fungi
as
a
source
for
antibacterial
compounds
Jawad
Anwar1,
Taj
Muhammad2,
Ulf
Göransson2,
Zafar
Iqbal1
1 Department of Agricultural Chemistry, University of Agriculture, 25100 Peshawar, Pakistan, 2 Division of
Pharmacognosy,
Department
of
Medicinal
Chemistry,
Uppsala
University,
Biomedical
Centrum
Box
574,
Husargatan
3,
SE
75123Uppsala,
Sweden
Phytopathogenic
bacteria
constantly
develop
resistance
against
existing
antibiotic
drugs.
To-‐
day,
this
is
a
real
threat
to
human
health,
but
it
also
leads
to
secondary
effects
in
agriculture
such
as
great
losses
of
crops
and
vegetable
yield.
Development
of
new
drugs,
especially
in
ar-‐
ea
of
infectious
diseases,
represents
today
one
of
the
most
important
area
of
research.
Natural
products
exhibit
biological
properties
including
antibacterial,
antifungal,
herbicidal,
anti-‐
cancer
and
insecticidal
which
are
used
for
pharmaceutical
drug
discovery
and
agro-‐chemical
applications
[1].
Fungi
are
known
to
produce
a
vast
array
of
secondary
metabolites
that
are
often
bioactive;
species
like
Aspergillus,
Penicillium,
Fusarium
and
Acrimonies
have
the
poten-‐
tial
to
produce
numerous
secondary
metabolites
[2].
In
the
current
work,
twelve
different
fungal
strains
of
both
habitats
(soil
borne
and
endophytes)
were
screened
against
two
phyto-‐
pathogenic
bacterial
strains,
i-‐e
Clavibacter
michiganensis
and
Xanthomonas
campestris,
using
dual
culture
technique
[3].
Based
on
the
antagonistic
effect
of
fungi
against
the
two
bacterial
strains,
three
fungi
named
Aspergillus
flavus,
Penicillium
EU003
and
Trichoderma
harzianum
were
selected.
The
cultivation
of
the
three
fungi
was
optimized
on
different
growth
media
with
the
aim
to
identify
the
medium
which
secondary
metabolites
possessing
comparatively
more
inhibition
against
phytopathogens
[4].
The
following
culture
media
were
used:
Glucose
Peptone
Yeast
Broth
(GPYB),
Yeast
Extract
Broth
(YEB),
Growth
Nutrient
Broth
(GNB)
and
Potato
Dextrose
Broth
(PDB),
and
the
fungi
were
then
extracted
with
ethyl
acetate.
All
organic
extracts
obtained
from
culture
media
were
tested
for
the
bench
top
antibacterial
activity
us-‐
ing
a
disc
diffusion
method
[5].
The
results
revealed
that
the
crude
extract
of
Aspergillus
flavus
grown
on
PDB
show
highest
inhibition
against
the
two
plant
pathogenic
bacterial
strains
as
compared
to
other
medias.
Similarly,
crude
extracts
of
Penicillium
EU003
and
Trichoderma
harzianum
showed
good
results
on
GNB
culture
media.
Acknowledgements: Higher Education Commission Pakistan
Keywords: Dual culture technique, culture media optimization, in-‐vitro antibacterial bioassay
References:
[1] Gershenzon
J,
Dudareva
N.
The
function
of
terpene
natural
products
in
the
natural
world.
Nature
Chem
Biol
2007;
3:
408-‐414
[2] Grabley
S,
Sattler
I.
Modern
Methods
of
Drug
Discovery.
Natural
products
for
lead
identifi-‐
cation:
Nature
is
a
valuable
resource
for
providing
tools
online
publication
2003
[3] Sonawane
A,
Mahajan
M,
Renake
S.
Antifungal
activity
of
a
fungal
isolates
against
pome-‐
granate
Wilt
pathogen
Fusarium.
Int
J
Curr
Microbiol
Appl
Sci
2015;
2:
48-‐57
[4] Joel
EL,
Bhimba
BV.
A
secondary
metabolite
with
antibacterial
activity
produced
by
man-‐
grove
foliar
fungus
Schizophyllum
commune.
IJCEBS
2013;
1:
2320-‐4087
[5] Xiulan
XU,
Miller
SA,
Gurel
B,
Gartemann
KH,
Eichenlaub
R,
Rajasheara
G.
Bioluminescence
imaging
of
Clavibacter
michiganensis
subsp.
michiganensis
infection
of
tomato
seeds
and
plants.
Appl
Environ
Biotech
2010;
76:
3978-‐3988
P636
Production
and
detection
of
the
natural
ionophore
Beauvericin
Jesús
M.
González1,
Amparo
Alfonso1,
MJ
Sainz
2,
Luis
M
Botana1
1Departamento
de
Farmacología,
Universidad
de
Santiago
de
Compostela,
Facultad
de
Veterinaria,
27002,
Lugo,
Spain,
2
Departamento
de
Producción
Vegetal,
Universidad
de
Santiago
de
Compostela,
Facultad
de
Veterinaria,
27002,
Lugo,
Spain.
Mycotoxins are secondary metabolites produced by several species of filamentous fungi.
Many of these metabolites have interesting therapeutic properties. Beauvericin is a cyclohex-‐
adepsipeptide mycotoxin produced by Fusarium species. This compound acts as an ionophore
by forming dimeric structures that can be crossed by ions. Ultra Performance Liquid Chroma-‐
tography coupled to Mass Spectometry-‐Ion Trap-‐Time of Flight (UPLC-‐MS-‐IT-‐ TOF) is one of
the best instruments for identifying new compounds, and it is a useful metabolite profiling
technology.
The aims of this work were for one side the identification of Fusarium strains capable of pro-‐
ducing Beauvericin, and for other side the optimization of an UPLC-‐MS-‐IT-‐TOF method for the
detection of this fungal metabolite. We tested twenty Fusarium strains which belong to fifteen
different Fusarium species. The strains were cultured over wheat, under controlled condi-‐
tions, during 15 days and then frozen until analysis. Once the samples were extracted and fil-‐
tered, they were analysed by UPLC-‐MS-‐IT-‐TOF. Exact mass of Beauvericin, m/z 784.4173 was
monitored and it was detected in seven strains. Beauvericin presence was confirmed by the
sodium and ammonium adducts in the mass spectra, and also by using an analytical standard
for final confirmation. The quantification of Beauvericin was done with a calibration curve
employing eight different concentrations (12.5-‐300 ng/mL). These results show that F. prolif-‐
eratum and F. anthophillum are suitable fungi species for Beauvericin production owing to the
great amount of this metabolite which they can generate, 34 and 36 µg per gram of wheat,
respectively. The other Fusarium species that produce this metabolite (F. foetens, F. ox-‐
ysporum, F. poae and F. sterilihyphosum) are able to produce among 1 and 18 µg of Beauver-‐
icin per gram of wheat. Besides, the UPLC-‐MS-‐IT-‐TOF method developed allows accurate and
reliable detection and quantification of natural compounds.
Acknowledgements:
Jesús
María
González
Jartín
is
supported
by
a
fellowship
from
Programa
de
For-‐
mación
de
Profesorado
Universitario
(FPU14/00166),
Ministerio
de
Educación,
Spain.
Keywords: Beauvericin, Fusarium, UPLC-‐MS-‐IT-‐TOF
P637
Peptaibols
from
Tichoderma
sp.
(MSX70741):
Isolation,
structure
elucidation
and
biological
activity
José
Rivera-‐Chávez1,
Huzefa
A.
Raja1,
Prashant
Metri2,
Ding
Xue2,
Cedric
J.
Pearce3,
Nicholas
H.
Oberlies1
1
Department
of
Chemistry
and
Biochemistry,
University
of
North
Carolina
at
Greensboro,
Greensboro,
NC
27402,
USA;
2Department
of
Molecular,
Cellular
and
Developmental
Biology,
University
of
Colorado,
Boulder,
CO
80309,
USA.,
3Mycosynthetix,
Inc.,
Hillsborough,
NC
27278,
USA.
Peptaibols
are
a
class
of
linear
peptides
with
a
high
content
of
α-‐aminoisobutyric
acid,
an
ac-‐
ylated
N-‐terminus,
while
the
C-‐terminus
may
consist
of
a
free
amino
acid,
2-‐amino
alcohol,
or
sugar
alcohol,
among
others
[1].
These
compounds
have
been
widely
studied
due
to
their
in-‐
teresting
biological
properties,
such
as
antibiotics,
anthelmintic
and
cytotoxic
agents
[2].
In
this
context,
MSX
70741
(Trichoderma
sp.),
a
peptaibols
producing
fungus
from
the
Mycosyn-‐
thetix
library
was
chemically
reinvestigated.
This
study
led
to
the
isolation
of
three
new
pep-‐
taibols
(1-‐3),
as
well
as
the
known
compounds
alamethicin
F50
(4),
alamethicin
II
(5),
atrovi-‐
ridin
J
(6),
and
trichobranchin
D-‐I
(7).
The
structures
of
the
isolates
were
established
using
a
set
of
spectroscopic
(1D
and
2D
NMR)
and
spectrometric
(HRESIMS/MSn)
techniques.
The
absolute
configuration
of
compounds
was
determined
by
Marfey’s
analysis
of
the
individual
amino
acids.
The
cytotoxic
activity
of
compounds
was
evaluated
in
an
MTT
assay
against
a
panel
of
cancer
cell
lines.
HTC
116
cell
line,
Atroviridin J
(6)
MSX 70741
Keywords:
Peptaibols,
1D
and
2D
NMR,
HRESIMS/MSn,
Biological
activity
References:
[1] Degenklob
T,
Berg
A,
Gams
W,
Schlegel
B,
Gräfe
U.
The
occurrence
of
peptaibols
and
struc-‐
turally
related
peptaibiotics
in
fungi
and
their
mass
spectrometric
identification
via
diag-‐
nostic
fragment
ions.
J
Pept
Sci
2003;
9:
666-‐678
[2] Ayers
S,
Ehrmann
BM,
Adcock
AF,
Kroll
DJ,
Carcache
de
Blanco
EJ,
Shen
Q,
Swanson
SM,
Falkinham
III
JO,
Wani
MC,
Mitchell
SM,
Pearce
CJ,
Oberlies
NH.
Peptaibols
from
two
uni-‐
dentified
fungi
of
the
order
Hypocreales
with
cytotoxic,
antibiotic,
and
anthelmintic
activ-‐
ities.
J
Pept
Sci
2012;
18:
500-‐510
P638
Development
of
microcystin
derivatives
as
novel
agents
against
OATP1B3-‐expressing
tumors
Julius
Krivec¹,
Urs
F.
Moschik1,
Wolfram
Lorenzen¹,
Julia
Moschny²,
Timo
H.
J.
Niedermeyer²
1Cyano
Biotech
GmbH,
Magnusstrasse
11,
12489
Berlin,
Germany;
2Interfaculty
Institute
of
Microbiology
and
Infection
Medicine,
University
of
Tübingen,
Auf
der
Morgenstelle
28,
72076
Tübingen,
Germany
Microcystins
(MC)
are
cyanobacterial
cyclic
heptapeptides.
They
potently
inhibit
the
eukary-‐
otic
phosphatase
families
PP1
and
PP2A
[1]
after
cellular
uptake
mediated
by
the
organic
ani-‐
on
transporting
polypeptides
OATP1B1
and
OATP1B3
[2].
These
transporters
are
mainly
ex-‐
pressed
in
liver
cells,
resulting
in
severe
liver
damage
upon
microcystin
exposure
[2].
Howev-‐
er,
OATP1B3
is
also
expressed
by
several
carcinoma
cells
from
other
organs
and
tissues
such
as
lung,
breast,
and
colon
[3].
Interestingly
the
hepatic
expression
levels
of
OATP1B3
lie
far
below
those
of
OATP1B1.
Thus,
microcystin
selectivity
toward
OATP1B3
should
result
in
de-‐
creased
hepatic
clearance
and
toxicity
as
well
as
in
increased
uptake
in
OATP1B3-‐expressing
tumor
cells,
creating
a
therapeutic
window
for
microcystins
as
anticancer
drugs.
Screening
23
microcystin
variants,
we
found
[D-‐Asp3,(E)-‐Dhb7]MC-‐HilR,
MC-‐RF
and
MC-‐RY
as
congeners
with
selectivity
for
OATP1B3.
Our
findings
suggest
that
an
arginine
residue
in
position
2
and
an
aromatic
amino
acid
in
position
4
favor
OATP1B3
selectivity
over
OATP1B1
[4].
In
order
to
generate
a
broader
spectrum
of
microcystins
with
these
structural
elements,
we
used
a
tech-‐
nique
known
as
precursor-‐directed
biosynthesis
[5].
Feeding
specific
unnatural
amino
acids
to
MC-‐LR
and
MC-‐YR
producing
strains
resulted
in
succesful
incorporation
of
these
precur-‐
sors.
Interestingly,
not
all
strains
capable
of
producing
MC-‐YR
were
also
able
to
incorporate
the
unnatural
amino
acid
derivatives.
Thus,
precursor
feeding
strategies
present
a
new
and
convenient
way
to
generate
novel
MC
variants
with
unusual
structural
features.
Keywords:
Microcystin,
anti-‐cancer
agents,
OATP1B3,
precursor-‐directed
biosynthesis
References:
[1] MacKintosh
C,
Beattie
KA,
Klumpp
S,
et
al.
Cyanobacterial
microcystin-‐LR
is
a
potent
and
specific
inhibitor
of
protein
phosphatases
1
and
2A
from
both
mammals
and
higher
plants.
FEBS
1990;
264:
187–192
[2] Fischer
WJ,
Altheimer
S,
Cattori
V,
et
al.
Organic
anion
transporting
polypeptides
ex-‐
pressed
in
liver
and
brain
mediate
uptake
of
microcystin.
Toxicol
Appl
Pharmacol
2005;
203:
257−263
[3] Liu
T,
Li
Q.
Organic
anion-‐transporting
polypeptides:
a
novel
approach
for
cancer
therapy.
J
Drug
Target
2014;
22:
14–22
[4] Niedermeyer
THJ,
Daily
A,
Swiatecka-‐Hagenbruch
M,
et
al.
Selectivity
and
Potency
of
Mi-‐
crocystin
Congeners
against
OATP1B1
and
OATP1B3
Expressing
Cancer
Cells.
PLoS
One
2014;
9:
e91476.
[5] Kennedy
J.
Mutasynthesis,
Chemobiosynthesis,
and
Back
to
Semi-‐Synthesis:
Combining
Synthetic
Chemistry
and
Biosynthetic
Engineering
for
Diversifying
Natural
Products.
Nat
Prod
Rep
2008;
25–34
P639
Bioguided
isolation
as
a
tool
for
the
discovery
of
novel
cosmeu-‐
ceutical
agents
from
microbial
biodiversity
Katerina
Georgousaki1,
Nikolaos
Tsafantakis1,
Sentiljana
Gumeni2,3,
Spiros
Fotinos2,
Ioannis
P.
Trougakos2,
Nikolas
Fokialakis1
1Department
of
Pharmacognosy
and
Natural
Products
Chemistry,
Faculty
of
Pharmacy,
University
of
Ath-‐
ens,
Panepistimioupolis,
15771,
Athens,
Greece,
2Department
of
Cell
Biology
and
Biophysics,
Faculty
of
Biology,
University
of
Athens,
Panepistimiopolis,
15784
Athens,
Greece,
3
Lavipharm
SA,
Agias
Marinas,
19002
Paiania
Attica,
Greece
In
the
frame
of
MICROSMETICS
EU
project
more
than
110
potential
candidate
fungi
and
acti-‐
nomycetes
strains
were
selected
to
be
studied.
In
total
more
than
1100
extracts
were
pro-‐
duced
and
a
broad
spectrum
of
bioassays
have
being
incorporated
for
the
evaluation
of
their
anti-‐ageing
activity,
including
analyses
for
anti-‐oxidant,
skin-‐protecting
and
skin-‐whitening
bioactivity.
Among
the
initial
1100
extracts,
100
were
selected
as
promising
bioactive
prod-‐
ucts
and
have
been
further
analyzed
by
LC-‐HRMS
for
profiling,
metabolomics
analysis
and
dereplication.
Through
metabolomics
analyses
the
20
most
interesting
extracts
(19
strains
under
14
conditions)
have
been
forwarded
for
large-‐scale
cultivation
and
for
confirmation
of
their
bioactivity.
In
total,
8
fungal
and
12
actinomycetes
extracts
were
further
investigated
for
their
antioxidant
(DPPH
and
ABTS
assays)
and
skin
whitening
(tyrosinase
assay)
activities;
in
parallel,
a
bioassays-‐guided
optimization
of
the
extraction
procedure
was
performed.
The
ten
most
active
extracts
have
been
further
evaluated
in
cell-‐based
assays
for
their
skin
whitening
(bleaching)
activity
(i.e.
tyrosinase
inhibition)
in
mouse
melanocytes
(B16F10
cell
line),
as
well
as
in
normal
human
skin
fibroblasts
for
their
capacity
to
activate
proteostasis
ensuring
anti-‐ageing
mechanisms,
namely
the
ubiquitin-‐proteasome
and
autophagy-‐
lysosomal
systems.
The
most
bioactive
extracts
have
been
selected
for
dereplication;
cultiva-‐
tion
in
larger
scale
and
isolation
of
bioactive
compounds.
The
dereplication
was
performed
using
UPLC-‐HRMS
while
the
isolation
of
the
compounds
contained
in
those
extracts
was
performed
using
both
classical
chromatographic
methods
(HPLC,
MPLC),
as
well
as
more
innovative
techniques,
like
SFC-‐MS
and
FCPC.
The
full
set
of
spectroscopic
data
(MS
and
NMR)
was
recorded
for
all
isolated
compounds
in
order
to
unam-‐
biguously
elucidate
their
structure.
Acknowledgements:
This
work
has
been
financially
supported
by
EU
under
the
frame
of
MICROSMETICS
project
(FP7-‐PEOPLE-‐IAPP
2013,
Grant
agreement:
612276).
Keywords: Cosmeceuticals, skin-‐whitening, anti-‐oxidant, fungi, actinomycetes
P640
Do
the
secondary
metabolites
from
Phanerochaete
chrysosporium
change
depending
on
their
growth
substrates?
Kirk
P.
Manfredi,
James
Humpal
Department of Chemistry and Biochemistry, University of Northern Iowa, Cedar Falls, Iowa USA
The
wood-‐rotting
fungus
P.
chrysosporium
has
been
extensively
studied
because
of
its
ability
to
degrade
lignin
as
well
as
a
wide
variety
of
environmentally
persistent
organic
pollutants
[1].
It
is
one
of
the
few
fungi
capable
of
digesting
lignin.
This
is
advantageous
to
the
fungus
because
it
allows
it
to
have
access
to
cellulose
and
hemicellulose
from
a
multitude
of
sources.
Several
lignin
peroxidases
and
manganese
peroxidases
are
responsible
for
the
biodegradation
of
lignin.
Many
environmentally
persistent
organic
compounds
are
oxidized
by
lignin
and
manganese
peroxidases.
Because
of
its
biodegradative
abilities,
there
is
substantial
interest
in
the
use
of
this
and
other
wood-‐rotting
fungi
for
bioremediation
[2].
Recent
genomic
studies
have
shown
that
P.
chrysosporium
also
contains
numerous
polyketide
synthases
and
non-‐
ribosomal
peptide
synthases
in
its
genome,
suggesting
its
potential
to
produce
useful
second-‐
ary
metabolites
[3].
Because
of
its
metabolic
versatility
and
its
ability
to
grow
on
a
vast
array
of
carbon
sources,
we
undertook
a
study
to
determine
if
its
production
of
secondary
metabo-‐
lites
varies
with
the
growth
substrate.
P.
chrysosporium
was
cultured
on
a
number
of
different
carbon
sources
(e.g.,
coffee,
saw
dust)
and
attempted
to
identify
some
marker
secondary
me-‐
tabolites
using
LC-‐MS.
Additionally,
we
selected
other
growth
substrates
(e.g.,
Petroselinum
crispum,
Ocimum
basilicum)
that
are
known
to
produce
an
array
of
natural
products
and
in-‐
vestigated
the
fate
of
these
compounds
during
the
organism’s
growth.
The
isolated
metabo-‐
lites
and
biotransformation
products
were
assayed
for
their
antimicrobial
activity.
Our
initial
results
have
shown
that
when
P.
chrysosporium
is
grown
on
coffee,
paper
or
rice
a
major
me-‐
tabolite
with
antimicrobial
properties
is
isolated.
Though
the
organism
grows
extremely
well
on
orange
peels,
lemon
peels,
and
Petroselinum
crispum
the
aforementioned
antimicrobial
compound
is
not
produced.
Further
structural
details
of
the
isolated
compounds
will
be
dis-‐
cussed.
Acknowledgements:
Professor
John
Bumpus
is
acknowledged
for
providing
P.
chrysosporium
and
his
ex-‐
pertise
in
its
growth.
Keywords: LC-‐MS, P. chrysosporium, secondary metabolites
References:
[1] Karas
P,
Perruchon
C,
Exrhou
K,
Ehallotis
C,
Karpouzas
D.
Potential
for
bioremediation
of
agro-‐industrial
effluents
with
high
loads
of
pesticides
by
selected
fungi.
Biodegradations
2011;
22:
215−228
[2] McGrath
R,
Singleton
I.
Pentachlorophenol
transformation
in
soil:
a
toxicological
assess-‐
ment.
Soil
Biol
Biochem
2000;
32:
1311−1314
[3] Martinez
D,
et
al.
Genome
sequence
of
lignocellulose
degrading
fungus
Phanerochaete
chrysosporium
strain
RP78.
Nat
Biotechnol
2004;
22:
695−700
P641
Chemotaxonomy
of
the
genus
Stemphylium
Kresten Jon Kromphardt Olsen, Birgitte Andersen
Department
for
Systems
Biology,
Søltofts
Plads,
Building
223,
Technical
University
of
Denmark,
DK-‐2800
Lyngby,
Denmark
The
filamentous
fungal
genus
Stemphylium
is
often
found
on
various
crops,
and
especially
the
common
animal
feed
plant
Medicago
sativa
(alfalfa)
is
often
infected
by
this
plant
pathogen.
Some
members
also
contaminate
food
products
such
as
tomatoes
and
cherries.
With
this
in
mind
it
is
important
to
consider
what
consequences
such
an
infection/contamination
can
have,
e.g.
production
of
mycotoxins.
Identification
of
such
pathogens
is
pivotal
to
determine
if
an
infection
is
to
be
further
handled
[1].
Traditionally
Stemphylium
isolates
have
been
identi-‐
fied
using
morphological
characters,
however,
correct
identification
takes
a
specialist.
Particu-‐
larly
S.
herbarum,
S.
botryosum,
S.
vesicarium,
S.
alfalfae
and
S.
gracilariae
have
very
similar
conidial
appearance
[3].
Phylogenetic
studies
using
DNA
sequencing
of
Stemphylium
have
shown
that
some
species
within
the
genus
are
distinguishable
via
phylogeny
(ITS,
GPD)
while
others
are
not
[4,
5].
Especially,
a
group
of
S.
herbarum,
S.
vesicarium,
S.
alfalfae,
S.
tomatonis
and
S.
sedicola
are
almost
identical
when
only
considering
phylogeny.
The
purpose
of
this
pro-‐
ject
was
to
utilize
current
chemical
analytical
methods,
to
build
a
profile
of
secondary
metabo-‐
lites
(chemotaxonomy),
to
attempt
to
distinguish
the
species
in
the
genus
Stemphylium.
The
previously
mentioned
species,
which
are
hard
to
distinguish
in
conventional
identification
methods,
were
of
special
interest.
A
total
of
90
isolates
were
analyzed
for
chemical
production
along
with
morphological
characters.
Some
of
the
detected
compounds
were
stemphol,
alter-‐
porriols
and
altersolanols,
of
which
the
last
two
are
shared
with
the
closely
related
genus
Al-‐
ternaria.
Both
datasets
(morphology
and
chemical
production)
were
clustered
to
produce
phenograms
of
the
isolates.
Some
isolates
from
previous
studies
[1,
2]
were
also
used
in
the
current
study
and
phylogenies
were
used
to
support
findings
in
the
current
study.
The
chemo-‐
taxonomy
did
not
have
a
strong
resolving
power
to
differentiate
the
species
in
question,
and
it
is
possible
that
the
previously
proposed
differences
in
morphology
are
not
a
proof
of
separate
species,
but
rather
variation
within
the
same
species.
Acknowledgements: Amy Rossman, USDA, for donation of Stemphylium isolates
Keywords: Chemotaxonomy, Stemphylium, HPLC-‐MS, mycotoxins, secondary metabolites
References:
[1] Frisvad
JC,
Andersen
B,
Thrane
U.
The
use
of
secondary
metabolite
profiling
in
chemotax-‐
onomy
of
filamentous
fungi.
Mycological
research
2008;
112:
231-‐240
[2] Simmons
EG.
Perfect
States
of
Stemphylium.
Mycologia
1969;
61:
1-‐26
[3] Simmons
EG.
Perfect
states
of
Stemphylium-‐IV.
Harvard
Papers
in
Botany
2001;
199-‐208
[4] Câmara
MPS,
O'Neill
NR,
Van
Berkum
P.
Phylogeny
of
Stemphylium
spp.
based
on
ITS
and
glyceraldehyde-‐3-‐phosphate
dehydrogenase
gene
sequences.
Mycologia
2002;
94:
660-‐
672
[5] Inderbitzin
P,
Yeshwant
RM,
Mary
LB.
Pleospora
species
with
Stemphylium
anamorphs:
a
four
locus
phylogeny
resolves
new
lineages
yet
does
not
distinguish
among
species
in
the
Pleospora
herbarum
clade.
Mycologia
2009;
101:
329-‐339
P642
Cytotoxic
activities
of
endophytic
fungi
extracts
from
Paepalan-‐
thus
planifolius:
A
Brazilian
evergreen
Marcelo
R.
de
Amorim1,
Weslei
B.
Botero1,
Ana
Caroline
Z.
Silva1,
Angela
R.
Araújo1,
Iracilda
Z.
Carlos2,
Lourdes
Campaner
dos
Santos1
1
Department
of
Organic
Chemistry,
Institute
of
Chemistry,
UNESP
–
Univ
Estadual
Paulista,
14800-‐900
Araraquara
–
SP,
Brazil,
2
Department
of
Biological
Sciences,
School
of
Pharmaceutical
Sciences,
UNESP
–
Univ
Estadual
Paulista,
14800-‐900
Araraquara
–
SP,
Brazil
The
Eriocaulaceae
family
comprises
approximately
1.200
species
divided
into
10
genera
[1].
Paepalanthus
has
approximately
357
species,
95%
of
which
are
endemic,
restricted
to
Brazil
and
many
of
these
are
endangered
[2].
Previous
studies
of
Paepalanthus
species
showed
the
presence
of
substances
with
biological
potential
such
as
cytotoxic
activity
[3].
Despite
the
large
number
of
studies
of
the
Eriocaulaceae
species,
there
are
few
studies
of
endophytic
fungi
associated
with
this
family,
which
demonstrates
the
need
for
researches
to
identify
the
chem-‐
ical
and
biological
composition
of
extracts
and
metabolites
produced
by
endophytic
fungi
as-‐
sociated
with
the
Paepalanthus
genus.
The
aim
of
this
work
is
to
evaluate
cytotoxic
activity
from
endophytic
fungi
extracts
isolated
from
Paepalanthus
planifolius
(Bong.)
Körn
using
MTT
assay.
In
this
study,
fifteen
endophytic
fungi
were
isolated
from
the
leaves,
capitula
and
scapes
of
P.
planifolius
and
were
inoculated
into
Potato
Dextrose
Broth
(PDB)
on
a
reduced
scale.
The
medium
was
autoclaved
and
afterwards
the
endophytes
were
inoculated
and
incu-‐
bated
in
static
mode
for
28
days.
The
flask-‐accumulated
mycelial
biomass
was
separated
from
the
aqueous
medium
by
filtration,
and
the
filtrate
was
subjected
to
liquid-‐liquid
partition
with
ethyl
acetate
(EtOAc).
The
EtOAc
fraction
was
evaporated
resulting
in
the
crude
extract
[4].
The
extracts
were
screened
and
evaluated
for
their
cytotoxic
activity;
the
in
vitro
growth
in-‐
hibitory
effect
of
the
extracts
were
compared
with
cisplatin
(positive
control)
using
the
MTT
assay
for
cells
lines
LM3
(mammary
adenocarcinoma)
and
LP07
(lung
adenocarcinoma),
and
for
human
cancer
cells
line
MCF-‐7
(breast
adenocarninoma)
[5].
The
corresponding
IC50
val-‐
ues
are
listed
in
Table
1.These
results
are
promising
in
our
search
for
new
bioactive
substanc-‐
es
from
endophytic
fungi
and
the
screening
will
be
used
to
select
the
bioactive
fungal
strains
in
subsequent
studies.
Table
1.
Cytotoxicity
values
for
extracts
Pp-‐F03
and
Pp-‐E02.
IC50
±
(SD)
[µg
mL-‐1]
Extracts
LM3
LP07
MCF-‐7
Pp-‐F03
68.25
±
1.02
110.17
±
4.90
31.68
±
0.46
Pp-‐E02
37.93
±
4.95
38.26
±
0.25
28.82
±
5.85
Cisplatin
9.09
±
1.11
1.3
±
0.12
5.88
±
1.38
Acknowledgements:
The
authors
would
like
to
thank
the
financial
support
provided
by
the
Fundação
de
Amparo
à
Pesquisa
do
Estado
de
São
Paulo
(FAPESP),
Grant
2015/04899-‐3,
awarded
to
L.C.S.,
and
M.R.A.
Would
like
to
thank
FAPESP
(2015/
11058-‐5)
for
the
scholarship
granted.
Keywords: Eriocaulaceae, endophytic fungi, MTT
References:
[1] Andrade
MJG,
Giulietti
AM,
Harley
RM,
van
den
Berg
C.
Blastocaulon
(Eriocaulaceae),
a
syn-‐
onym
of
Paepalanthus:
morphological
and
molecular
evidence.
Taxon
2011;
60:
178-‐184
[2] Costa
FN,
Trovó
M,
Sano
PT.
Eriocaulaceae
na
Cadeia
do
Espinhaço:
riqueza,
endemismo
e
ameaças.
Megadiversidade
2008;
4:
89-‐97
[3] Devienne
KF,
Raddi
MSG,
Varanda
EA,
Vilegas
W.
In
vitro
cytotoxicity
of
some
natural
and
semi-‐synthetic
isocoumarins
from
Paepalanthus
bromelioides.
Z
Naturforsch
C
2002;
57:
85-‐88
[4] Chapla
VM,
Zeraik
ML,
Leptokarydis
IH,
Silva
GH,
Bolzani
VS,
Young
MCM,
Pfenning
LH,
Araújo
AR.
Antifungal
compounds
produced
by
Colletotrichum
gloeosporioides,
an
endo-‐
phytic
fungus
from
Michelia
champaca.
Molecules
2014;
19:
19243-‐19252
[5] Rocha
F
V,
Barra
C
V,
Mauro
A
E,
Carlos
I
Z,
Nauton
L,
El
Ghozzi
M,
Gautier
A,
Morel
L,
Netto
A
V
G.
Synthesis,
characterization,
x-‐ray
structure,
DNA
cleavage,
and
cytotoxic
activities
of
Palladium(II)
complexes
of
4-‐phenyl-‐3-‐thiosemicarbazide
and
triphenylphosphane.
Eur
J
Inorg
Chem
2013;
4499-‐4505
P643
Influence
of
the
medium
and
preferred
cereal
substrate
on
sec-‐
ondary
metabolite
production
by
species
from
Penicillium
series
Viridicata
Magnus
Hallas-‐Møller,
Kristian
F.
Nielsen
and
Jens
C.
Frisvad
Department
of
Systems
Biology,
Technical
University
of
Denmark,
Søltofts
Plads
building
221,
2800
Kgs.
Lyngby,
Denmark
Stored
cereals
are
vulnerable
to
mycotoxin
contamination
by
the
major
fungal
genus
Penicilli-‐
um
and
especially
the
species
from
the
series
Viridicata
which
is
constituted
by
the
7
species:
P.
aurantiogriseum,
P.
cyclopium,
P.
freii,
P.
neoechinulatum,
P.
polonicum,
P.
tricolor,
and
P.
viridicatum,
but
also
known
cereal
associated
species
such
as
P.
verrucosum
and
P.
hordei
[1].
These
species
have
been
chemically
investigated
[2],
but
often
only
by
now
outdated
analyti-‐
cal
systems
like
thin-‐layer
chromatography
or
HPLC-‐DAD.
On
top
of
this,
their
secondary
me-‐
tabolite
profiles
have
routinely
only
been
screened
on
laboratory
agar
media
such
as
yeast
extract
sucrose
(YES)
or
czapek
yeast
extract
agar
(CYA),
so
our
knowledge
on
how
constitu-‐
tively
the
known
secondary
metabolites
are
expressed
on
their
natural
substrates
such
as
e.g.
whole
wheat
grains,
is
limited.
Based
on
the
mentioned
species
we
will
here
present
a
thor-‐
ough
investigation
on
the
exo-‐metabolome,
performed
on
a
powerful
UHPLC-‐HRQTOFMS
platform,
which
coupled
together
with
our
in-‐house
MSMS
library
on
fungal
secondary
me-‐
tabolites
allowed
to
identify
several
known,
but
new
to
these
species,
secondary
metabolites
like
e.g.
the
ergot
alkaloid
pathway
found
in
Claviceps
purpurea
and
Penicillium
commune
[3]
or
Chrysogine,
which
is
produced
by
various
phylogenetically
unrelated
ascomycetous
genera,
were
found
in
three
species
from
the
series
Viridicata.
Acknowledgements:
We
would
like
to
thank
the
Novo
Nordisk
Foundation
grant
#NNF
130
C0005201
for
the
funding
of
this
study.
We
are
grateful
to
Agilent
Technologies
for
the
Thought
Leader
Donation
of
the
UHPLC-‐MS
systems.
Keywords: Pencillium, cereals, UHPLC-‐HRQTOFMS, mycotoxins
References:
[1] Mills
JT,
Seifert
KA,
Frisvad
JC,
Abramson
D.
Nephrotoxigenic
Penicillium
species
occurring
on
farm-‐stored
cereal
grains
in
western
Canada.
Mycopathologia
1995;
130:
23-‐28
[2] Frisvad
JC,
Smedsgaard
J,
Larsen
TO,
Samson
RA.
Mycotoxins,
drugs
and
other
extrolites
produced
by
species
in
Penicillium
subgenus
Penicillium.
Stud
Mycol
2004;
49:
201-‐241
[3] Gerhards,
N,
Neubauer
L,
Tudzynski
P,
Shu-‐Ming
L.
Biosynthetic
Pathways
of
Ergot
Alka-‐
loids.
Toxins
2014;
6:
3281-‐3281
P644
New
diketomorpholines
from
a
facultative
marine-‐derived
As-‐
pergillus
sp.
(ACA-‐9)
Manuel
A.
Aparicio-‐Cuevas1,
Isabel
Rivero-‐Cruz1,
María
C.
González2,
Huzefa
A.
Raja3,
Mario
Figueroa1
1
Departamento
de
Farmacia,
Facultad
de
Química,
and
2
Instituto
de
Biología,
Universidad
Nacional
Autónoma
de
México,
Mexico
City
04510,
Mexico,
3
Department
of
Chemistry
and
Biochemistry,
University
of
North
Carolina
at
Greensboro,
Greensboro,
North
Carolina
27402,
United
States
As
part
of
our
continuing
search
for
novel
bioactive
fungal
compounds,
the
facultative
marine-‐
derived
Aspergillus
sp.
(ACA-‐9)
was
isolated
from
a
transition
zone
between
land
and
the
sea
at
the
Acapulco
Bay,
Guerrero,
Mexico
[1].
Fungal
identification
was
performed
based
on
the
morphology
of
reproductive
structures,
protein-‐coding
genes
(RPB2
and
β-‐tubulin)
sequenc-‐
ing,
as
well
as
nuclear
ribosomal
internal
transcribed
spacer
(ITS)
barcoding
[2].
Chemical
analysis
of
the
organic
(CHCl3-‐MeOH)
extract
from
the
axenic
solid
(moisture
rice)
culture
yielded
five
new
diketomorpholine
(DKM)
derivatives
together
with
shornephine
A.
Their
structures
were
elucidated
using
a
set
of
spectroscopic
(400,
500,
and
700
MHz
NMR)
and
spectrometric
(UPLC-‐ESIHRMS-‐MS/MS)
techniques,
along
with
chemical
degradations
and
derivatizations.
In
addition,
these
compounds
were
tested
for
antiproliferative
activity
against
colon
(HCT-‐15),
cervix
(HeLa),
and
breast
(MCF-‐7
and
MDA-‐MB-‐231)
carcinoma
cell
lines,
and
for
antimicrobial
activity
using
an
array
of
bacteria
(Escherichia
coli,
Pseudomonas
aeruginosa,
Staphylococcus
aureus,
and
Bacillus
subtilis)
and
fungi
(Candida
albicans
and
Aspergillus
terre-‐
us).
To
the
best
of
our
knowledge,
this
is
the
second
report
of
natural
products
with
a
DKM
scaffold
[3].
Acknowledgements:
This
research
was
supported
by
grant
CB-‐2014
236564
(to
M.F.)
from
the
Consejo
Nacional
de
Ciencia
y
Tecnología
(CONACyT),
Mexico
City,
Mexico.
MA.A.-‐C.
acknowledges
a
fellowship
from
CONACyT
(274048)
to
pursue
graduate
studies.
Keywords: Diketomorpholines, fungi, Aspergillus, bioactive compounds
References:
[1] Velez
P,
González
MC,
Rosique-‐Gil
E,
Cifuentes
J,
Reyes-‐Montes
MR,
Capello-‐García
S,
Hanlin,
RT.
Community
structure
and
diversity
of
marine
ascomycetes
from
coastal
beaches
of
the
southern
Gulf
of
Mexico.
Fungal
Ecol
2013;
6:
513-‐521
[2] Samson
RA,
Visagie
CM,
Houbraken
J,
Hong
SB,
Hubka
V,
Klaassen
CH,
Perrone
G,
Seifert
KA,
Susca
A,
Tanney
JB,
Varga
J.
Phylogeny,
identification
and
nomenclature
of
the
genus
Aspergillus.
Stud
Mycol
2014;
78:141-‐173
[3] Khalil
ZG,
Huang
XC,
Raju
R,
Piggott
AM,
Capon
RJ.
Shornephine
a:
structure,
chemical
sta-‐
bility,
andP-‐glycoprotein
inhibitory
properties
of
a
rare
diketomorpholine
from
an
Aus-‐
tralian
marine-‐derived
Aspergillus
sp.
J
Org
Chem
2014;
79:
8700-‐8705
P645
Evaluation
of
lichen
compounds
as
inhibitors
of
Candida
albicans
biofilms
Marion
Girardot1,
Marion
Millot2,
Clément
Bernard1,
Lengo
Mambu2,
Christine
Imbert1
1UMR
CNRS
7267
Laboratory
of
Ecology
and
Biology
of
Interactions,
Faculty
of
Medicine
Pharmacy,
Uni-‐
versity
of
Poitiers,
Bât
D1,
6
rue
de
la
Milétrie,
TSA
51115,
86073
Poitiers
cedex
9,
France;
2EA
1069
La-‐
boratory
of
Chemistry
of
Natural
Substances,
Faculty
of
Pharmacy,
University
of
Limoges,
2
rue
du
Dr
Marcland,
87025
Limoges
cedex,
France
C.
albicans,
a
frequent
commensal
coloniser
of
the
human
gastrointestinal
tract,
can
also
be-‐
come
an
opportunistic
fungal
pathogen
especially
thanks
to
its
capacity
to
form
biofilms.
This
organisation
confers
to
Candida
a
1,000-‐fold
greater
resistance
to
certain
antifungal
drugs
[1].
In
this
context
of
frequent
infections
involving
Candida
biofilms
often
resistant
to
current
treatments,
new
anti-‐biofilm
substances
are
needed.
Lichens
are
symbiotic
associations
be-‐
tween
an
alga
or/and
a
cyanobacterium
and
a
fungus
and
represent
a
reservoir
of
compounds
with
therapeutic
potential
[2].
In
this
study,
screening
of
fifty
lichen
extracts
was
performed
against
sessile
C.
albicans
cells
by
evaluating
their
anti-‐adherent
activity
(against
a
2h
biofilm)
using
a
XTT
method.
Seven
lichen
extracts
had
significant
anti-‐adherent
activity
(p<0.05)
at
concentrations
<
10
μg/mL.
Their
activity
was
then
tested
against
24h
preformed
biofilms
and
results
were
analysed
using
three
methods
(XTT,
CFU
counts
and
trypan
blue
staining)
in
order
to
evaluate
their
influence
on
both
fungal
metabolism,
cultivability
and
viability.
All
ex-‐
tracts
demonstrated
anti-‐biofilm
activity
using
concentrations
<
10
μg/mL,
especially
Evernia
prunastri,
Ramalina
fastigiata
and
Xanthoparmelia
tinctina,
whose
activity
persisted
at
48h
of
biofilm
treatment.
CFU
tests
demonstrated
a
drastic
decrease
of
the
amount
of
yeast
cells
con-‐
stituting
the
biofilm
after
treatment
(inhibition
≥
80%).
Trypan
blue
staining
showed
that
cells
growing
as
biofilms
were
still
alive
after
treatment,
suggesting
a
non
lethal
effect
of
the
tested
products.
This
work
suggested
a
dispersant
or
removing
action
not
targeting
the
fungal
cell
membrane.
HPLC
and
TLC
profiles
of
the
active
extracts
revealed
some
similarities
in
chemical
content.
The
main
compounds
were
isolated
from
these
extracts
by
precipitation
or
by
chromatographic
methods
and
identified
by
NMR
and
MS.
Among
them,
the
depsides
ever-‐
nic,
squamatic
and
thamnolic
acids
displayed
anti-‐adherent
activity
(IC50
≤
25µg/mL)
whereas
the
depside
atranorin
and
depsidones
were
inactive.
Only
thamnolic
acid
demonstrated
activ-‐
ity
against
preformed
biofilms.
Thus,
the
preventive
and
curative
potential
of
lichen
metabo-‐
lites
was
demonstrated
against
C.
albicans
biofilms.
Acknowledgements:
CNRS
is
acknowledged
for
financial
support
(Licafilm
project
/
Exomod
PEPS
re-‐
search
program
2014,
2015)
Keywords: lichens, depside, biofilm, Candida albicans
References:
[1] Herwald
SE,
Kumamoto
CA.
Candida
albicans
niche
specialization:
features
that
distin-‐
guish
biofilm
cells
from
commensal
cells.
Curr
Fungal
Infect
Rep
2014;
8:
179−184
[2] Zambare
VP,
Christopher
LP.
Biopharmaceutical
potential
of
lichens.
Pharm
Biol
2012;
50:
778−798
P646
Screening
of
actinobacteria
for
inhibition
of
quorum
sensing
of
Chromobacterium
violaceum
CV026
and
Staphylococcus
aureus
PC322
Nico
Ortlieb,
Timo
H.
J.
Niedermeyer
Interfaculty
Institute
of
Microbiology
and
Infection
Medicine,
Eberhard
Karls
Universität
Tübingen,
Auf
der
Morgenstelle
28,
72076
Tübingen,
Germany
and
German
Center
for
Infection
Research,
partner
site
Tübingen,
Germany
The
spread
of
antibiotic
drug
resistances
and
the
occurrence
of
multidrug
resistant
bacteria
are
one
of
the
major
threats
of
the
21th
century,
as
they
lead
to
infections
that
cannot
be
treat-‐
ed
adequately.
Interference
with
bacterial
virulence
or
biofilm
formation
by
inhibition
of
bac-‐
terial
quorum
sensing
(QS)
has
been
discussed
as
new
strategy
to
overcome
resistances
[1].
This
is
seen
as
a
promising
strategy
since
bacterial
growth
is
not
affected
and
thus
emergence
of
resistance
is
not
very
likely
[2].
Although
actinobacteria
potentially
produce
compounds
that
inhibit
QS,
only
few
studies
have
been
reported
[3].
Therefore,
about
1000
strains
of
the
Tübinger
actinobacteria
strain
collection
have
been
screened
for
inhibition
of
quorum
sensing
using
the
reporter
strains
Chromobacterium
violaceum
CV026
and
Stahylococcus
aureus
PC322
[4]
after
the
development
of
a
medium-‐throughput
agar
plug
assay
based
screening
system.
56
Streptomyces
sp.
strains
were
found
to
inhibit
violacein
production
in
Chromobac-‐
terium
violaceum
CV026.
Furthermore,
41
Streptomyces
sp.
strains
showed
reduced
expres-‐
sion
of
hla
in
the
monitor
strain
Staphyloccocus
aureus
PC322.
A
HPLC-‐DAD-‐MS
based
chemi-‐
cal
screening
of
the
20
most
active
strains
revealed
3
strains
producing
compounds
that
could
not
be
dereplicated
using
commercial
and
in-‐house
databases
of
natural
products.
In
order
to
isolate
bioactive
compounds,
extracts
of
these
strains
have
been
subjected
to
bioactivity
guid-‐
ed
fractionation
using
several
chromatographic
techniques.
The
structures
of
these
com-‐
pounds
have
been
elucidated
mainly
by
nuclear
magnetic
resonance
spectroscopy
and
tan-‐
dem
high
resolution
mass
spectrometry.
Our
screening
highlights
the
potential
of
actinobac-‐
teria
to
produce
compounds
interfering
with
the
quorum
sensing
of
other
bacteria,
a
potential
bioactivity
of
actinobacterial
specialized
metabolites
that
has
long
been
neglected.
Keywords:
Quorum
sensing
inhibition,
Chromobacterium
violaceum
CV026,
Staphylococcus
aureus
PC322,
Actinobacteria,
natural
product
research
References:
[1] Clatworthy
AE,
Pierson
E,
Hung
DT.
Targeting
virulence:
a
new
paradigm
for
antimicrobial
therapy.
Nat
Chem
Biol
2007;
3:
541-‐548
[2] Gerdt
JP,
Blackwell
HE.
Competition
studies
confirm
two
major
barriers
that
can
preclude
the
spread
of
resistance
to
quorum-‐sensing
inhibitors
in
bacteria.
ACS
Chem
Biol
2014;
9:
2291-‐2299
[3] Polkade
AV,
Mantri
SS,
Patwekar
UJ,
Jangid
K.
Quorum
Sensing:
An
Under-‐Explored
Phe-‐
nomenon
in
the
Phylum
Actinobacteria
Front
Microbiol
2016;
7:
131
[4] Nielsen
A,
Nielsen
KF,
Frees
D,
et
al.
Method
for
screening
compounds
that
influence
viru-‐
lence
gene
expression
in
Staphylococcus
aureus.
Antimicrob
Agents
Chemother
2010;
54:
509-‐512
P647
Bioactive
type
A
proanthocyanins
from
fungus
Laurobasidium
lauri
João
Serina1,
Maria
J.
Carvalho2,
Tatiana
Weinhold1,
Paula
C.
Castilho1
1
Centro
de
Química
da
Madeira,
Faculty
of
Exact
Sciences
and
Engineering,
Madeira
University,
Campus
Penteada,
9000-‐390,
Funchal,
Portugal,
2
Faculty
of
Exact
Sciences
and
Engineering,
Madeira
University,
Campus
Penteada,
9000-‐390,
Funchal,
Portugal
Laurobasidium
lauri,
commonly
known
as
Madre
de
Louro,
is
a
parasitic
fungus
of
Laurus
trees
which
has
a
large
use
in
local
folk
medicine,
usually
as
an
alcoholic
tincture
[1].
In
this
study,
the
antioxidant
and
antimicrobial
activities
of
several
extracts
of
Madre
de
Louro
were
evaluated
and
enzymatic
studies
were
performed
using
in-‐vitro
assays
[2].
The
ethanol
extract
revealed
higher
activity
in
all
assays.
Results
also
showed
the
ethanol
extract
to
be
a
potent
inhibitor
of
α-‐glucosidase,
compared
to
positive
control
acarbose.
IC50
values
were
1.04
µg/mL
for
the
methanol
extract
and
IC50
3.43
µg/mL
for
acarbose.
Alcoholic
extracts
exert
antibacterial
activity
against
Staphylococcus
aureus,
with
a
MIC
of
62.5
µg/mL.
Thus,
a
LC-‐MSn
technique
was
applied
to
determine
the
main
compounds
in
this
extract,
revealing
catechin
monomers,
dimers
and
trimers,
with
special
relevance
for
A-‐type
procyanidins.
Molecular
docking
was
performed
in
an
attempt
to
identify
the
compounds
which
exerted
the
observed
inhibitory
effects.
However,
per
se,
none
of
the
analysed
compounds
had
a
binding
affinity
close
to
acarbose
(-‐15.6
kcal/mol)
in
the
same
docking
conditions,
but
a
combination
of
two
ligands
leading
to
synergistic
interactions
is
proposed
as
responsible
for
the
enhanced
activity
observed
in
vitro.
Acknowledgements:
The
Portuguese
Foundation
for
Science
and
Technology
(FCT)
is
acknowledged
for
the
strategic
project
PEst-‐OE/QUI-‐674/UI0674/2014
Keywords: Laurobasidium lauri, procyanidins, LC-‐MSn, α-‐glucosidase, molecular docking
References:
[1] Tavares L, Carrilho D, Tyagi M, Barata D, Serra AT, Duarte CMM, Duarte RO, Feliciano RP,
Bronze MR, Chicau P, Espírito-‐Santo MD, Ferreira RB, dos Santos CN. Antioxidant capacity
of Macaronesian traditional medicinal plants. Molecules 2010; 15: 2576-‐2592
[2] Gouveia-‐Figueira SC, Gouveia CA, Carvalho MJ, Rodrigues AI, Nording ML, Castilho PC. Anti-‐
oxidant capacity, cytotoxicity and antimycobacterial activity of Madeira archipelago en-‐
demic Helichrysum dietary and medicinal plants. Antioxidants 2014; 3: 713-‐729
P658
Modulation
of
a
lichen-‐associated
fungus
for
improved
biosyn-‐
thesis
of
bioactive
secondary
metabolites
Peter
M.
Eze1,
Blessing
O.
Umeokoli2,
Huiqin
Chen3,
Zhen
Liu3,
Festus
B.C.
Okoye2,
Charles
O.
Esimone1,
Peter
Proksch3
1
Department
of
Pharmaceutical
Microbiology
and
Biotechnology,
Faculty
of
Pharmaceutical
Sciences,
Nnamdi
Azikiwe
University,
PMB
5025
Awka,
Anambra
State.
Nigeria.
2
Department
of
Pharmaceutical
and
Medicinal
Chemistry,
Faculty
of
Pharmaceutical
Sciences,
Nnamdi
Azikiwe
University,
PMB
5025
Awka,
Anambra
State.
Nigeria.
3
Institute
of
Pharmaceutical
Biology
and
Biotechnology,
Heinrich
Heine
University,
40225
Duesseldorf,
Germany.
In
this
study,
Apiospora
montagnei
isolated
from
a
lichen-‐Cladonia
sp
was
modulated
to
im-‐
prove
its
biosynthesis
of
bioactive
secondary
metabolites.
A
solid
state
fermentation
of
A.
montagnei
was
carried
out
on
rice
medium
for
21
days
at
22°C
and
the
fungal
secondary
me-‐
tabolites
were
extracted
using
ethyl
acetate.
The
crude
extract
was
subjected
to
several
chromatographic
separation
methods
and
the
resulting
fractions
were
subjected
to
HPLC-‐
DAD
analysis.
Detected
compounds
were
then
isolated
and
characterized
using
various
tech-‐
niques
such
as
NMR
spectroscopy,
MS
analysis,
etc.
The
metabolic
profile
of
A.
montagnei
was
then
established.
Manipulation
of
cultural
conditions
of
the
fungus
by
addition
of
carbon
and
nitrogen
sources,
and
also
several
sodium
salts;
cultivation
of
the
fungus
at
different
tempera-‐
tures;
as
well
as
co-‐cultivation
with
Bacillus
subtilis
and
Streptomyces
lividans,
respectively,
were
carried
out
to
improve
the
biosynthesis
of
identified
bioactive
compounds.
Several
bio-‐
active
compounds
which
were
undetected
or
those
detected
at
very
low
levels
in
the
initial
crude
extract
of
A.
montagnei
were
up-‐regulated
after
manipulation
of
the
cultural
conditions
of
the
fungus.
These
compounds
include-‐Anomalin
B,
Anomalin
A,
Norlichexanthone,
N-‐
hydroxy-‐apiosporamide,
Apiosporamide,
Decarboxycitrinone,
Decarboxyhydroxycitrinone,
Myrocin
A,
and
8-‐hydroxy-‐3-‐methyl-‐9-‐oxo-‐9H-‐xanthene-‐1-‐carboxylic
acid
methyl
ether.
Culti-‐
vation
of
fungus
at
28oC,
co-‐cultivation
with
B.
subtilis,
and
the
addition
of
various
sodium
salts,
nitrogen,
and
carbon
sources
triggered
expression
of
silent
fungal
genes.
Addition
of
NaCl
(5%)
and
NH4Cl
(1%),
as
well
as
co-‐cultivation
with
S.
lividans,
induced
the
production
of
new
compounds.
Results
of
this
study
indicate
that
co-‐cultivation,
as
well
as
manipulation
of
cultural
conditions,
can
result
in
increased
yields
of
previously
described
metabolites
and
previously
undetected
metabolites.
Keywords: Apiospora montagnei, Cladonia sp., modulation, co-‐cultivation, secondary metabo-‐
lites.
P649
Screening
of
cyanobacteria
extracts
for
inhibitory
activity
against
the
cysteine
protease
rhodesain
of
Trypanosoma
brucei
Ronja
Kossack¹,
Trang
Nguyen¹,
Steffen
Breinlinger1,
Tanja
Schirmeister²,
Timo
H.
J.
Nieder-‐
meyer¹
¹
Interfaculty
Institute
of
Microbiology
and
Infection
Medicine,
Eberhard
Karls
Universität
Tübingen,
Auf
der
Morgenstelle
28,
72076
Tübingen,
Germany
and
German
Center
for
Infection
Research,
partner
site
Tübingen,
Germany;
²
Institute
of
Pharmacy
und
Biochemistry,
Johannes
Gutenberg-‐Universität
Mainz,
Staudinger
Weg
5,
55128
Mainz,
Germany
Cyanobacteria
are
known
as
a
rich
source
of
structurally
diverse
protease
inhibitors
[1],
but
no
screening
for
rhodesain
inhibitors
has
been
reported
yet.
Therefore
a
collection
of
about
670
cyanobacteria
extracts
was
screened
for
inhibitory
activity
against
rhodesain
according
to
Breuning
[2].
The
trypanosomal
cysteine
protease
rhodesain
plays
a
major
role
during
parasitic
infection
by
Trypanosoma
brucei,
known
as
human
African
trypanosomiasis
(HAT)
[3].
Rhodesain
is
involved
in
the
parasitic
crossover
of
the
blood-‐brain-‐barrier,
leading
to
the
late-‐stage
of
HAT
[4]
and
in
the
synthesis
of
variant
surface
glycoproteins
(VSGs)
of
trypanosomes,
enabling
T.
brucei
to
elude
host
immune
response
[5].
Thus
it
is
regarded
as
a
promising
target
for
the
development
of
urgently
needed
new
therapies
[3].
The
screening
of
the
extract
library
revealed
12
extracts
from
the
genera
Aphanizomenon,
Cylindrospermum,
Fischerella,
Nostoc,
and
Oscillatoria
with
a
rhodesain
inhibition
of
higher
than
90%
at
0.1
mg/mL.
The
most
promising
hit
extract
from
a
Nostoc
strain
has
subsequently
been
fractionated
using
flash
chromatography
and
preparative
high-‐performance
liquid
chromatography
in
order
to
isolate
the
active
compounds.
Five
structurally
related
compounds
(Mr
603
to
62
g/Mol)
with
rhodesain
inhibition
of
more
than
90%
at
0.1
mg/mL
could
be
isolated
in
addition
to
three
less
active
compounds
of
the
same
compound
family.
Interestingly,
the
most
abundant
compound
of
this
compound
family
is
not
active
in
the
assay.
Structure
elucidation
of
the
active
compounds
has
been
carried
out
mainly
by
nuclear
magnetic
resonance
spectroscopy
and
tandem
high
resolution
mass
spectrometry.
Our
study
confirms
that
cyanobacteria
present
a
valuable
source
for
protease
inhibitors.
Further
investigations
concerning
activities
against
other
proteases
relevant
in
infectious
diseases
are
planned.
Keywords: Natural products, cyanobacteria, rhodesain, human African trypanosomiasis
References:
[1]
Radau
G.
Cyanopeptides:
A
New
and
Nearly
Inexhaustible
Natural
Resource
for
the
Design
and
Structure-‐Activity
Relationship
Studies
of
New
Inhibitors
of
Trypsin-‐like
Serine
Proteases.
Curr
Enz
Inhib
2005;
1:
295-‐307
[2]
Breuning
A,
Degel
B,
Schulz
F,
et
al.
Michael
Acceptor
Based
Antiplasmodial
and
Antitrypanosomal
Cysteine
Protease
Inhibitors
with
Unusual
Amino
Acids.
J
Med
Chem
2010;
53:
1951-‐1963
[3]
Ettari
R,
Tamborini
L,
Angelo
IC,
et
al.
Inhibition
of
Rhodesain
as
a
Novel
Therapeutic
Modality
for
Human
African
Trypanosomiasis.
J
Med
Chem
2013;
56:
5637-‐5658
[4]
Lonsdale-‐Eccles
JD,
Grab
DJ.
Trypanosome
hydrolases
and
the
blood–brain
barrier.
Trends
Parasitol
2002;
18:
17-‐19
[5]
Overath
P,
Chaudhri
M,
Steverding
D,
Ziegelbauer
K.
Invariant
surface
proteins
in
bloodstream
forms
of
Trypanosoma
brucei.
Parasitol
Today
1994;
10:
53-‐58
P650
Mensacarcin,
a
soil
derived
polyketide,
exhibits
strong
universal
anti-‐proliferative
effects
in
human
cancer
cell
lines
and
induces
cell
death
selectively
in
melanoma
cells.
Birte Plitzko, Sandra Loesgen
Department of Chemistry, Oregon State University, 153 Gilbert Hall, 97331 Corvallis OR, USA
Mensacarcin
is
a
unique,
highly
oxygenated
polyketide
that
was
isolated
from
a
soil
derived
Streptomyces
bottropensis
[1].
The
in
vitro
NCI
60
cancer
cell
line
panel
revealed
a
potent
anti-‐
proliferative
response
and
‘‘COMPARE-‐negative’’
profile
indicative
of
a
unique
mechanism
of
action.
Here
we
report
its
potent
cytostatic
effects
(GI:
0.5
μM)
in
various
cancer
cell
viability
assays
and
its
selective
cytotoxicity
against
melanoma
cell
lines
(LC50:
1.0
μM).
We
show
that
mensacarcin
induces
morphologically
and
biochemically
distinct
forms
of
cell
death
according
to
cell
type.
SK-‐Mel-‐28
melanoma
cells
show
strong
chromatin
condensation
and
procaspase-‐
3
and
PARP
cleavage
upon
treatment
with
mensacarcin,
effectively
inducing
apoptotic
cell
death.
TZM-‐Bl
cervix
cancer
cells
show
a
similarly
strong
response
to
mensacarcin,
in
which
cytostasis
is
accompanied
by
fast
progression
into
cell
death
via
apoptotic
and
secondary
ne-‐
crosis.
Interestingly,
HCT-‐116
colon
carcinoma
cells
treated
at
similar
concentrations
of
men-‐
sacarcin
are
strongly
inhibited
in
growth
but
do
not
show
signs
of
cell
death.
Cell
cycle
analy-‐
sis
reveals
that
mensacarcin
can
arrest
cells
at
any
given
stage
eventually
leading
into
apopto-‐
sis.
The
mode
of
action
of
mensacarcin
was
further
explored
on
the
molecular
level.
We
found
evidence
for
interaction
with
DNA
using
intercalating
studies,
and
we
examined
its
potential
to
directly
bind
to
DNA
and
single
nucleotides
using
the
Comet
assays,
ITC,
NMR
and
MS.
Mensacarcin
Acknowledgements: Prof. Axel Zeeck for providing mensacarcin, OSU start up funds, NSF IOS-‐1557804
Keywords: Soil bacteria, Streptomyces, Apoptosis, Cytotoxicity, Melanoma Cells.
References:
[1] Maier
S,
Pflüger
T,
Loesgen
S,
Asmus
K,
Brötz
E,
Paululat
T,
Zeeck
A,
Andrade
S,
Bechthold
A.
Novel
insights
into
the
bioactivity
and
epoxide
formation
in
mensacarcin
by
MsnO8.
ChemBioChem
2014;
15:
749-‐756
P651
Chemical
investigations
of
the
fruiting
bodies
of
Pleurotus
cornu-‐
copiae
and
biological
activities
of
the
isolated
compounds
Seoung
Rak
Lee,
Seulah
Lee,
Hee
Jeong
Eom,
Hee
Rae
Kang,
Jae
Sik
Yu,
Tae
Kyoung
Lee,
Jiwon
Baek,
Dahae
Lee,
Won
Se
Suh,
Ki
Hyun
Kim
School
of
Pharmacy,
Sungkyunkwan
University,
Suwon
440-‐746,
Republic
of
Korea
Pleurotus
cornucopiae
(Pleurotaceae)
is
an
edible
mushroom
widely
distributed
in
Korea,
Japan,
China,
Siberia,
and
North
America
[1].
The
mushroom
is
traditionally
known
for
its
use
as
health
functional
food
because
of
its
various
bioactivities
including
immunoregulation,
fa-‐
tigue
resistance,
and
antioxidant
and
antitumor
effects
[2].
Previous
studies
reported
signifi-‐
cant
bioactive
constituents
from
P.
cornucopiae,
such
as
D-‐mannitol,
lectin,
and
laccase
[3].
Recently,
a
phytochemical
study
of
the
mycelia
of
this
mushroom
reported
the
isolation
of
monoterpenoids
and
sesquiterpenoids
exhibiting
inhibitory
activity
toward
NO
production
[4].
In
this
study,
we
report
the
isolation
of
one
new
nucleoside
analogue
(1),
together
with
six
known
compounds
(2-‐7).
The
structures
of
isolated
compounds
were
determined
by
analyz-‐
ing
spectroscopic
data
(1H,
13C
NMR,
2D
NMR,
MS)
and
comparison
with
the
literature
data.
Although
compound
3
was
previously
reported
as
a
synthetic
product,
it
was
isolated
for
the
first
time
from
natural
products.
Compounds
4-‐7
were
isolated
for
the
first
time
from
P.
cor-‐
nucopiae.
All
the
isolated
compounds
from
this
mushroom
were
tested
for
their
effects
on
an-‐
giogenesis
in
HUVECs
and
cisplatin-‐induced
nephrotoxicity
in
LLC-‐PK1
cells.
OH NH2
N N N
N O
N N H3CO
N N OH NH2
OH O OH
O OH H3CO N
HO
HN OH OH
1 2 3 4
H H H
OH H
HO HO OH
OH HO
OH OH OH
OH OH
5 6 7
Acknowledgements:
This
research
was
supported
by
Basic
Science
Research
Program
through
the
Na-‐
tional
Research
Foundation
of
Korea
(NRF)
funded
by
the
Ministry
of
Science,
ICT
&
Future
Planning
(2015R1C1A1A02037383)
Keywords: Pleurotus cornucopiae, Pleurotaceae, Nucleoside analogue, Structural elucidation
References:
[1] Jang
JH,
Jeong
SC,
Kim
JH,
Lee
YH,
Ju
YC,
Lee
JS.
Characterisation
of
a
new
antihypertensive
angiotensin
I-‐converting
enzyme
inhibitory
peptide
from
Pleurotus
cornucopiae.
Food
Chem
2001;
127:
412−418
[2] Petr
V,
Vendula
V,
Vera
M,
Jiri
G.
Degradation
of
lignocellulose
by
Pleurotus
ostreatus
in
the
presence
of
copper,
manganese,
lead
and
zinc.
Res
Microbiol
2005;
156:
670−676
[3] Oguri
S,
Ando
A,
Nagata
Y.
A
novel
developmental
stage-‐specific
lectin
of
the
basidiomy-‐
cete
Pleurotus
cornucopiae.
J
Bacteriol
1996;
178:
5692−5698
[4] Wang
S,
Bao
L,
Zhao
F,
Wang
Q,
Li
S,
Ren
J,
Li
L,
Wen
H,
Guo
L,
Liu
H.
Isolation,
identifica-‐
tion,
and
bioactivity
of
monoterpenoids
and
sesquiterpenoids
from
the
mycelia
of
edible
mushroom
Pleurotus
cornucopiae.
J
Agric
Food
Chem
2013;
61:
5122−5129
P652
Inhibitory
effect
of
isolated
constituents
from
sclerotia
of
Poria
cocos
on
LPS-‐induced
NO
production
Seoung
Rak
Lee1,
Seulah
Lee1,
Hee
Jeong
Eom1,
Hee
Rae
Kang1,
Jae
Sik
Yu1,
Tae
Kyoung
Lee1,
Jiwon
Baek1,
Dahae
Lee1,
Won
Se
Suh1,
Ki
Hyun
Kim1
1School
of
Pharmacy,
Sungkyunkwan
University,
Suwon
440-‐746,
Republic
of
Korea
Poria
cocos
Wolf,
well
known
as
‘Fu-‐Ling’,
is
a
medicinal
mushroom
of
the
family
Polyporace-‐
ae,
which
is
widely
distributed
in
Korea,
China,
and
East
Asian
countries
[1].
This
mushroom
is
widely
utilized
in
traditional
medicine
for
sedative,
diuretic,
and
tonic
effects
[2].
Recently,
the
sclerotia
of
P.
cocos
showed
potential
as
a
natural
drug
for
improving
diuresis,
treatment
of
invigorating
spleen,
and
stabilizing
the
mind
[3,4].
Previous
phytochemical
studies
reported
that
more
than
50
triterpenoids
were
isolated
from
P.
cocos,
and
most
of
them
were
lanos-‐
tane-‐type
triterpenoids
[5].
In
our
continuing
search
for
bioactive
constituents
from
Korean
wild
mushrooms,
we
investigated
the
EtOH
extract
of
the
sclerotia
of
P.
cocos
in
the
course
of
bioassay-‐guided
isolation.
The
EtOH
extract
was
solvent-‐partitioned
with
n-‐hexane,
CH2Cl2,
EtOAc,
and
n-‐BuOH.
We
found
that
the
CH2Cl2
fraction
showed
significant
inhibitory
activity
on
nitric
oxide
(NO)
production
in
LPS-‐induced
RAW
264.7
macrophage
cells.
Phytochemical
investigation
of
the
CH2Cl2
fraction
led
to
the
isolation
of
six
triterpenoids
(1-‐6),
one
lignan
(7),
and
one
phenolic
compound
(8).
All
the
isolated
compounds
were
structurally
identified
by
spectroscopic
methods
(1H-‐NMR,
13C-‐NMR,
MS)
and
comparison
with
previously
reported
spectroscopic
data.
Anti-‐inflammatory
activities
of
the
isolated
compounds
were
determined
as
potential
regulators
for
excessive
inflammatory
response
in
RAW
264.7
macrophage
cells.
HOOC HOOC HOOC HOOC
H H H H
OH OH OH
HOOC O
O O HO
H H H H
1 2 3 4
HO
HOOC HOOC
O H
H H
H H3CO O
O OCH3
H
OH
HO HO OH OH
H H
5 6 7 8
Acknowledgements:
This
research
was
supported
by
Basic
Science
Research
Program
through
the
Na-‐
tional
Research
Foundation
of
Korea
(NRF)
funded
by
the
Ministry
of
Science,
ICT
&
Future
Planning
(2015R1C1A1A02037383)
Keywords: Poria cocos, Polyporaceae, Triterpenoid, Nitric oxide
References:
[1] Ling
Y,
Chen
MC,
Wang
K,
Sun
ZL,
Li
ZX,
Wu
B,
Huang
CG.
Systematic
screening
and
charac-‐
terization
of
the
major
bioactive
components
of
Poria
cocos
and
their
metabolites
in
rats
by
LC-‐ESI-‐MS.
Biomed
Chromatogr
2012;
26:
1109-‐1117
[2] Rios
JL.
Chemical
constituents
and
pharmacological
properties
of
Poria
cocos.
Planta
Med
2011;
77:
681-‐691
[3] Kobira
S,
Atsumi
T,
Kakiuchi
N,
Mikage
M.
Difference
in
cultivation
characteristics
and
ge-‐
netic
polymorphism
between
Chinese
and
Japanese
strains
of
Wolfiporia
cocos
Ryvarden
et
Gilbertson
(Poria
cocos
Wolf).
J
Nat
Med
2012;
66:
493-‐499
[4] Wang
YZ,
Li
T,
Zhao
YL,
Zhang
J,
Liu
HG.
Contents
of
some
metabolites
in
the
peel
and
flesh
of
the
medicinal
mushroom
Wolfiporia
cocos
(F.A.
Wolf)
Ryvarden
et
Gilb.
Int
J
Med
Mushrooms
2012;
14:
79-‐83
[5] Rios
JL,
Andujar
I,
Recio
MC,
Giner
RM.
Lanostanoids
from
fungi:
a
group
of
potential
anti-‐
cancer
compounds.
J
Nat
Prod
2012;
75:
2016-‐2044
P653
Marine
fungi:
A
novel
source
of
cosmeceutical
applications
Shivankar
Agrawal1,
Sunil
Kumar
Deshmukh1,
Colin
Barrow2,
Alok
Adholeya1*
1
TERI-‐Deakin
Nano
Biotechnology
Centre,
Biotechnology
and
Management
of
Bioresources
Division,
The
Energy
and
Resources
Institute,
New
Delhi
110003,
India.
2
Centre
for
Chemistry
and
Biotechnology
(CCB),
School
of
Life
and
Environmental
Sciences,
Deakin
University,
Pigdons
Road,
Waurn
Ponds,
Victo-‐
ria
3216,
Australia
A
variety
of
genetic
and
environmental
factors
cause
various
cosmetics
and
dermatological
problems.
There
are
already
claimed
drugs
available
in
market
for
treating
these
problems.
However,
the
challenge
remains
in
finding
more
potent,
environmental
friendly,
causing
min-‐
imal
side
effects
and
economical
cosmeceuticals.
This
leads
to
an
increased
demand
for
natu-‐
ral
cosmeceutical
products
in
the
last
few
decades.
Plant
derived
ingredients
are
limited
be-‐
cause
plants
either
contain
toxic
metabolites,
grow
too
slow
or
seasonal
harvesting
is
a
prob-‐
lem.
The
research
work
carried
out
in
this
project
aims
at
isolation,
characterization
of
marine
fungal
secondary
metabolite
and
evaluating
their
potential
use
in
future
cosmetic
skin
care
products.
We
have
isolated
and
purified
70
morphologically
different
fungal
isolates
from
var-‐
ious
marine
habitats
of
the
India.
These
isolates
have
been
functionally
characterised
for
anti-‐
tyrosinase,
antioxidant
and
anti-‐acne
activities.
For
molecular
characterisation,
the
Internal
Transcribed
spacer
(ITS)
region
of
15
functionally
active
marine
fungal
isolates
was
amplified
using
universal
primers,
ITS1
and
ITS4
and
sequenced.
Out
of
15
marine
fungal
isolates
crude
extract
of
strains
D4
and
P2
showed
70%
and
57%
tyrosinase
inhibition
at
1mg/mL
respec-‐
tively.
Strain
D5
has
showed
significant
inhibition
against
Propionibacterium
acnes
and
Staph-‐
ylococcus
epidermidis.
In
addition,
all
these
strains
also
displayed
DPPH-‐
radical
scavenging
activity
and
may
be
utilized
as
a
skin
cosmeceutical
applications
purification
and
characteri-‐
zation
of
crude
extracts
for
identification
of
active
lead
molecule
is
under
process.
Acknowledgements:
The
authors
thank
TERI
and
Deakin
University
for
providing
necessary
infrastruc-‐
ture
and
financial
support.
Keywords: Marine Fungi, cosmeceutical, anti-‐tyrosinase, anti-‐acne.
P654
Indole
alkaloids
from
Penicillium
genus:
Identification,
isolation,
structure
elucidation
and
cytotoxicity
Svetlana
A
Kalinina1,2,
Annika
Jagels1,
Benedikt
Cramer1,
Hans-‐Ulrich
Humpf1,2
1Institute
of
Food
Chemistry,
Westfälische
Wilhelms-‐Universität
Münster,
Corrensstraße
45,
48149
Mün-‐
ster,
Germany;
2NRW
Graduate
School
of
Chemistry,
Westfälische
Wilhelms-‐Universität
Münster,
Wil-‐
helm-‐Klemm-‐Str.
10,
48149
Münster,
Germany
Fungi
are
major
plant
and
insect
pathogens,
but
they
are
not
nearly
as
important
as
agents
of
disease
in
vertebrates.
Growth
of
fungi
on
animal
hosts
is
a
reason
of
diseases
collectively
called
mycoses,
while
dietary,
respiratory,
dermal,
and
other
exposures
to
toxic
fungal
metab-‐
olites
lead
to
the
diseases
called
mycotoxicoses
[1,2].
There
are
still
few
well-‐characterized
fungi
and
most
of
these
fungi
produce
several
unknown
metabolites,
thus
making
fungi
a
promising
source
for
the
discovery
of
new
lead
compounds.
The
genus
Penicillium
contains
many
toxigenic
species
(approximately
100),
and
the
range
of
mycotoxin
classes
produced
by
this
genus
is
much
broader
than
that
of
any
others.
Penicillium
toxins
can
be
divided
into
two
wide
groups
based
on
toxic
effect:
those
that
effect
liver
and
kidney
function
and
those
that
are
neurotoxins.
The
most
important
of
them
are:
ochratoxin
A,
citrinin,
penicillic
acid,
citre-‐
oviridin,
penitrems,
roquefortines,
auranthine,
aurantiamine
and
cyclopiazonic
acid.
In
the
course
of
our
study
of
toxins
from
Penicillium
spp.,
penitrems
were
successfully
isolated.
Also,
effects
of
various
stress-‐factors
(oxidative
stress,
salinity,
nitrogen
and
carbon
sources,
pH,
temperature
etc.)
on
the
fungal
growth
and
production
of
the
metabolites
were
analyzed.
New
rapid
and
effective
methods
of
extraction
and
isolation
of
these
toxins
were
developed.
Thus,
fungal
cultures
were
extracted
with
acetonitrile,
followed
by
separation
of
compounds
using
flash
chromatography
(normal-‐phase
column
SNAP-‐25)
with
a
cyclohexane/ethyl
acetate
gradient.
Final
purification
of
alkaloids
was
carried
out
with
preparative
HPLC
using
isocratic
mode
of
appropriate
solvent.
The
structures
of
isolated
compounds
were
elucidated
by
NMR
and
HPLC-‐HRMS
experiments. Newly
obtained
mycotoxins
were
screened
for
their
cytotoxici-‐
ty
against
Hep
G2
cells
using
the
Cell
Counting
Kit-‐8
(CCK-‐8)
assay
with
IC50
=
46
μM
for
peni-‐
trem
A.
Penitrems
showed
moderate
cytotoxic
effect.
H
H
H 2C O H
H 2C O R1
R1 OH R1
O
OH OH
OH
N
R H N CH3
O R H O
Penitrem A, R=Cl, R1=OH Penitrem C, R=Cl, R1=H CH 2
Penitrem B, R=R1=H Penitrem D, R=R1=H
Penitrem E, R=H, R1=OH
Penitrem G, R=H, R1=OH
Penitrem F, R=Cl, R1=H
Keywords: Penicillium spp., penitrems, indole-‐based alkaloids, CCK-‐8
References:
[1] Bennett
JW,
Klich
M.
Mycotoxins.
Clin
Microbiol
Rev
2003;
3:
497–516
[2] Turner
NW,
Subrahmanyam
S,
Piletsky
SA.
Analytical
methods
for
determination
of
myco-‐
toxins:
a
review.
Anal
Chim
Acta
2009;
632:
168−180
P655
Activation
of
fungal
secondary
metabolism
by
disturbance
of
the
two-‐component
signal
transduction
system
and
identification
of
the
nectriapyrone
biosynthetic
gene
cluster
Takayuki
Motoyama,
Yoko
Tanaka,
Hiroyuki
Osada
RIKEN CSRS, 2-‐1 Hirosawa, Wako-‐shi, Saitama 351-‐0198, Japan
Filamentous
fungi
have
been
a
rich
source
of
secondary
metabolites
for
drug
development.
After
whole
genome
sequencing
analyses,
it
has
been
revealed
that
filamentous
fungi
have
much
more
secondary
metabolism
genes
than
expected.
It
suggests
that
most
secondary
me-‐
tabolism
genes
are
poorly
expressed
under
laboratory
conditions.
Secondary
metabolite
pro-‐
duction
might
be
strictly
regulated
to
produce
under
specific
environmental
conditions.
We
predicted
that
secondary
metabolite
production
would
be
activated
by
disturbance
of
signal
transduction
pathways
involved
in
environmental
responses.
Here,
we
show
that
secondary
metabolite
production
can
be
induced
by
disturbance
of
the
two-‐component
signal
transduc-‐
tion
system
[1]
in
the
rice
blast
fungus
Magnaporthe
oryzae,
a
model
plant
pathogenic
fungus.
We
constructed
knockout
strains
for
a
MAP
kinase
gene
(OSM1)
that
works
downstream
of
the
two-‐component
system.
The
Δosm1
strains
showed
enhanced
production
of
tenuazonic
acid
biosynthesized
by
a
fungal
NRPS–PKS
hybrid
enzyme
[2].
Next,
we
constructed
MoYPD1
gene
knockout
strains
of
the
Δosm1
strains.
MoYPD1
codes
for
a
His-‐containing
phosphotrans-‐
fer
(HPt)
protein
essential
for
phospho-‐relay
in
the
fungal
two-‐component
system.
The
Δosm1
ΔMoypd1
strains
showed
induced
production
of
two
polyketide
compounds,
nectriapyrone
(1)
and
its
new
hydroxylated
analog
(2).
In
M.
oryzae,
production
of
nectriapyrone
has
not
been
reported
yet.
Nectriapyrone
is
known
as
a
secondary
metabolite
produced
by
some
fungi
including
endophytes.
DNA
microarray
analysis
showed
that
expression
of
a
PKS
gene
is
in-‐
duced
in
the
Δosm1
ΔMoypd1
strain.
Gene
knockout
of
this
PKS
gene
indicated
that
this
gene
is
essential
for
nectriapyrone
biosynthesis.
A
closely
located
O-‐methyltransferase
gene
was
also
involved
in
nectriapyrone
biosynthesis,
indicating
that
there
is
a
nectriapyrone
biosyn-‐
thetic
gene
cluster.
O O
HO
O O O O
nectriapyrone (1) hydroxynectriapyrone (2)
Acknowledgements: This study was supported in part by Grants-‐in-‐Aid from the Ministry of Education,
Culture, Sports, Science and Technology of Japan and by the Science and Technology Research Promotion
Program for Agriculture, Forestry, Fisheries and Food Industry.
Keywords: Magnaporthe oryzae, secondary metabolite, gene cluster activation, endophyte
References:
[1] Motoyama
T,
Ochiai
N,
Morita
M,
Iida
Y,
Usami
R,
Kudo
T.
Involvement
of
putative
re-‐
sponse
regulator
genes
of
the
rice
blast
fungus
Magnaporthe
oryzae
in
osmotic
stress
re-‐
sponse,
fungicide
action,
and
pathogenicity.
Curr
Genet
2008;
54:
185-‐195
[2] Yun
CS,
Motoyama
T,
Osada
H.
Biosynthesis
of
the
mycotoxin
tenuazonic
acid
by
a
fungal
NRPS–PKS
hybrid
enzyme.
Nature
Commun
2015;
6:
8758.
P656
Prediction
of
secondary
metabolite
encoding
genes
based
on
chemical
structure
analysis
Thomas
Isbrandt1,
Maria
Lund
Nielsen1,
Casper
Hoeck3,
Yuksel
Gezgin4,
Rasmus
J.
N.
Frand-‐
sen2,
Kristian
Fog
Nielsen1,
Thomas
Ostenfeld
Larsen1
1
Natural
Product
Discovery,
Department
of
Systems
Biology,
Technical
University
of
Denmark,
Søltofts
Plads,
Building
221.
2
Eucaryotic
Molecular
Cell
Biology,
Department
of
Systems
Biology,
Technical
Uni-‐
versity
of
Denmark,
Søltofts
Plads,
Building
223.
3
Department
of
Chemistry,
Technical
University
of
Den-‐
mark,
Kemitorvet,
Building
207.
4
Ege
Universities,
Bornova-‐Izmir,
Turkey.
Dereplication
of
the
secondary
metabolite
profile
from
the
filamentous
fungus
Aspergillus
brasiliensis,
by
High
Performance
Liquid
Chromatography
coupled
with
Diode
Array
Detection
and
High
Resolution
Mass
Spectrometry,
[1]
lead
to
the
discovery
of
a
novel
biomarker
having
a
unique
UV
spectrum
and
elemental
composition.
Structural
elucidation
based
on
Nuclear
Magnetic
Resonance
spectroscopy
of
the
pure
compound
revealed
an
apolar
polyketide
or
fatty
acid
derived
secondary
metabolite,
possibly
assembled
from
two
entities,
a
C8
and
a
C12
chain,
fused
via
a
Claisen-‐like
condensation
and
subsequent
cyclisation
to
form
a
core
lactone
ring
structure.
The
compound
has
been
named
brasenol.
To
our
knowledge,
only
two
other
compunds
contain
a
similar
ring
structure;
namely
alternaric
acid
from
Alternaria
solani
and
fujikurin
D
found
in
Fusarium
fujikuroi
[2,3].
Despite
the
apolar
nature
of
the
compound
initial
bioassay
investigation
have
demonstrated
antibacterial
activity
(MIC=28.44
µg/mL)
against
methicillin-‐resistant
Staphylococcus
aureus
MB5393.
We
are
currently
investigating
whether
other
closely
related
black
Aspergilli
in
section
nigri
could
also
be
identified
as
brasenol
pro-‐
ducers.
This
will
be
followed
by
comparative
bioinformatics
analysis
of
their
already
se-‐
quenced
genomes,
setting
the
scene
for
construction
of
knock
out
mutants
of
potential
candi-‐
date
genes
involved
using
CRISPR/Cas9
in
A.
brasiliensis
[4].
This
poster
will
summarize
our
efforts
towards
characterization
of
the
biosynthetic
pathway
of
this
new
compound.
Figure
2.
Structure
of
brasenol.
Acknowledgements:
We
thank
Fundación
Medina,
Granada,
Spain
for
bioactivity
assessment.
Keywords: HPLC-‐DAD-‐HRMS, dereplication, Aspergillus brasiliensis, CRISPR/Cas9, biomarker
References:
[1] Nielsen
KF,
Månsson
M,
Rank
C,
Frisvad
JC,
Larsen
TO.
Dereplication
of
microbial
natural
products
by
LC-‐DAD-‐TOFMS.
J
Nat
Prod
2011;
74:
2338-‐2348
[2] Brian
PW,
Curtis
PJ,
Hemming
HG,
Unwin
CH,
Wright
JM.
Alternaric
acid,
a
biologically
ac-‐
tive
metabolic
product
of
the
fungus
Alternaria
solani.
Nature
1949;
164:
534-‐534
[3] Von
Bargen
KW,
Niehaus
E-‐M,
Krug
I,
Bergander
K,
Würthwein
E-‐U,
Tudzynski
B,
Humpf
H-‐
U.
Isolation
and
structure
elucidation
of
fujikurins
A–D:
Products
of
the
PKS19
gene
clus-‐
ter
in
Fusarium
fujikuroi.
J
Nat
Prod
2015;
78:
1809-‐1815
[4] Nødvig
CS,
Nielsen
JB,
Kogle
ME,
Mortensen
UH.
A
CRISPR-‐Cas9
system
for
genetic
engi-‐
neering
of
filamentous
fungi.
PLoS
One
2015;
10:
e0133085
P657
Chemical
and
biological
investigation
of
Algerian
lichens
Rafika
Brakni1,
Monia
Serradj1,
Xavier
Fernandez2,
Thomas
Michel2
1Department
de
biologie,
Université
de
Badji
Mokhtar
Annaba,
Laboratoire
de
biologie
végétale
et
envi-‐
ronnement.
BP
12
23000.
Annaba,
Algérie.
2
Institut
de
Chimie
de
Nice,
UMR
CNRS
7272,
Université
Nice
Sophia
Antipolis,
Parc
Valrose,
06108
Nice
CEDEX
2,
France
Lichens
are
symbioses
association
between
alga
and/or
a
cyanobacteria
and
a
fungus
in
self-‐
sustaining
partnerships
in
which
each
plays
a
well-‐defined
role:
algae
synthesize
organic
mat-‐
ter
(first
metabolism)
and
fungal
partner
produce
a
variety
of
secondary
metabolites
exhibit-‐
ing
various
biological
activities
such
as
antiviral,
anti-‐tumor,
anti-‐inflammatory,
analgetic,
antiproliferative,
antibacterial
[1,2].
In
Algeria
it
exists
a
very
diverse
lichen
flora
including
some
species
(Ramalina
farinacea,
fla-‐
voparmelia
caperata)
used
only
as
bio
indicator
of
air
pollution
[3,4].
Lichens
are
especially
abundant
in
the
east
region
of
Annaba,
which
is
characterized
by
cork
oak
forest,
which
sup-‐
ports
one
of
the
highest
levels
of
biodiversity.
However,
no
research
has
been
conducted
yet
to
evaluate
the
chemical
and
biological
qualities
of
this
lichenic
wealth.
In
this
purpose,
the
aim
of
this
research
work
was
to
explore
chemical
diversity
and
biological
activities
of
four
lichen
species
from
Algeria
Cladonia
rangiformis,
Roccella
phycopsis,
Ramalina
fastigeata
and
R.
farinacea.
This
study
reports
preliminary
results
concerning
the
evaluation
of
Algerian
lichen
as
cosmet-‐
ic
agents.
Indeed,
a
high
throughput
screening
was
performed
on
enzyme
involved
in
skin
disorders
(tyrosinase,
elastase,
lipoxygenase)
as
well
as
on
antioxidant
potential
of
lichens.
R.
phycopsis
was
active
against
tyrosinase
(48.89%)
and
lipoxygenase
(64.87%)
whereas
R.
fari-‐
naceae
showed
the
highest
anti
elastase
(87.80%)
and
antioxidant
potential
(46.32%).
Using
UPLC-‐HRMS
analyses
we
were
able
to
identify
huge
number
of
lichenic
compounds
such
as
orsellinic
acid,
montgnetol,
erythrin
(R.
phycopsis),
evernic
acid
(R.
fastigeata),
usnic
acid
(R.farinaceae
and
R.
fastigeata)
and
rangiformic
acid
from
C.
rangiformis.
Keywords:
Lichen
metabolites,
tyrosinase,
lipoxygenase,
elastase,
UPLC-‐HRMS.
References:
[1] Huneck
S.
New
results
on
the
chemistry
of
lichen
substances
in
progress
in
the
chemistry
of
organic
natural
products.
Springer
2001;
86:
1-‐276.
[2] Halama
P,
Haluwin
C.
Antifungal
activity
of
lichen
extracts
and
lichenic
acids.
BioControl
2004;
49:
95-‐107.
[3] Semadi
A.
Lead
pollution
monitoring
by
transplanted
lichens
in
Annaba
area
(Algeria).
Pollution
atmosphérique
1993;
90:
86-‐102
[4] Serradj
M,
Ahmed
A,
Boumedris
ZE,
Djebar
MR,
Tahar
A.
Réponses
d’antioxydants
chez
Flavoparmelia
caperata
(L.)
Hale
à
la
pollution
atmosphérique
au
niveau
de
deux
zones
urbaine
et
semi-‐urbaine
dans
la
région
d’Annaba
(Est
de
l’Algérie).
Pollution
atmosphé-‐
rique,
2014,
n°221
P658
Insects-‐entomopathogens
interactions
to
discover
new
insecti-‐
cidal
and
antimicrobial
compounds
Seinde
Touré1,
I.
Dusfour2,
D.
Stien3,
V.
Eparvier1
1
CNRS,
Institut
de
Chimie
des
Substances
Naturelles
(ICSN),
1
avenue
de
la
terrasse,
91198
Gif-‐sur-‐Yvette
Cedex,
France,
2
Unité
d’entomologie
médicale,
Institut
Pasteur
de
la
Guyane,
23
Avenue
Pasteur,
BP
6010,
97306
Cayenne
Cedex,
3
Sorbonne
Universités,
UPMC
Univ
Paris
06,
CNRS,
Laboratoire
de
Biodiversité
et
Biotechnologies
Microbiennes
(LBBM),
Observatoire
Océanologique,
66650
Banyuls-‐sur-‐mer,
France
The
search
of
insecticidal
and
antimicrobial
molecules
represents
a
global
challenge
for
public
health
[1].
On
one
hand,
vector-‐borne
diseases
remain
an
important
burden
while
the
efficacy
of
existing
insecticides
is
decreasing
due
to
the
development
of
resistances.
On
the
other
hand,
most
of
drugs
available
against
dermatophytes,
yeast
or
bacteria
suffer
of
drawbacks,
emergence
of
resistant/multiresistant
strains
and
toxicity.
Entomopathogenic
microorganisms
have
deleterious
effects
on
insects
due
to
their
develop-‐
ment
into
the
body
and
insecticidal
compounds
they
can
excrete.
During
the
process
of
colo-‐
nization,
they
most
likely
produce
active
compounds
against
other
microbial
species,
which
in
turn
protect
insects
by
mutualistic
association
[2].
Thus,
it
is
reasonable
to
postulate
that
in-‐
teractions
between
insects
and
insect
pathogenic
microorganisms
are
governed
by
specific
insecticides
and/or
antimicrobial
metabolites.
In
order
to
better
understand
microbial
competitions
on
insect
and
eventually
isolate
active
secondary
metabolites,
we
undertook
to
isolate
microbial
species
from
live
infected
insects.
Ethyl
acetate
extracts
of
the
57
isolated
strains
have
been
tested
on
Aedes
aegypti
mosquito
larvae
and
on
human
microbial
pathogens
(Staphyloccocus
aureus,
methicillin-‐resistant
S.
au-‐
reus,
Candida
albicans
and
Trichophyton
rubrum).
Cytotoxicity
bioassays
have
been
carried
out
on
three
cell
lines
(MRC5:
normal
human
lung
tissue,
KB:
human
cervical
cancer
cells,
MDA-‐231:
metastasic
melanoma).
High
hit
rate
was
recorded,
with
11,
7,
and
9
extracts
displaying
significant
insecticide,
anti-‐
microbial,
and
cytotoxic
potential,
respectively.
Up
to
now,
14
secondary
metabolites
have
been
isolated.
Acknowledgements:
This
work
has
benefited
from
an
“Investissement
d’Avenir”
grant
managed
by
Agence
Nationale
de
la
Recherche
(CEBA,
ref
ANR-‐10-‐LABX-‐0025).
Keywords: Entomopathogens, insecticide, Aedes aegypti, antimicrobial
References:
[1] WHO.
Fact
sheet
N°387.
(2014).
Available
at
http://www.who.int/mediacentre/factsheets/fs387/en/.
Accessed
February
2016
[2] Nirma
C,
Eparvier
V,
Stien
D.
Antifungal
Agents
from
Pseudallescheria
boydii
SNB-‐CN73
Isolated
from
a
Nasutitermes
sp.
termite.
J
Nat
Prod
2013;
76:
988-‐991
P659
New
antibacterial
small
molecules
from
insect-‐associated
bacte-‐
ria
in
Bombyx
mori
and
Nicrophorus
concolor
Yern-‐Hyerk Shin1, Ki-‐Bong Oh2, Jongheon Shin1, Dong-‐Chan Oh1
1 Natural Products Research Institute, College of Pharmacy, Seoul National University, 1, Gwanak-‐ro,
Gwanak-‐gu,
Seoul
08826,
Republic
of
Korea
2
Department
of
Agricultural
Biotechnology,
College
of
Agri-‐
culture
&
Life
Science,
Seoul
National
University,
1,
Gwanak-‐ro,
Gwanak-‐gu,
Seoul
08826,
Republic
of
Korea
Investigation
of
secondary
metabolites
of
bacteria
associated
with
eukaryotic
hosts
has
be-‐
come
a
promising
approach
to
discover
novel
bioactive
compounds
[1].
Especially,
insect-‐
associated
microbes
have
been
highlighted
for
a
new
source
of
natural
products
because
of
their
tremendous
biological
and
chemical
diversity
[2,3].
In
this
study,
we
isolated
bacterial
strains
from
silkworms
(Bombyx
mori)
and
burying
beetles
(Nicrophorus
concolor).
Chemical
analysis
of
the
bacteria
indicated
that
a
couple
of
actinobacterial
strains
produced
new
anti-‐
bacterial
small
molecules.
Chromatographic
isolation
and
spectroscopic
analysis
of
the
new
compounds
led
us
to
dis-‐
cover,
piceamycin
(1)
and
three
of
new
macrocyclic
lactams
(SD53.449,
SD53.419A,
and
SD53.419B.
2-‐4),
derived
from
silkworm-‐associated
bacterial
strain,
Streptomyces
sp.
(#SD53)
[4].
The
planar
structures
of
the
macrocyclic
lactams
were
determined
by
spectro-‐
scopic
analysis
of
1D
&
2D
NMR,
MS,
and
UV
data.
The
relative
and
absolute
configurations
of
the
lactams
were
established
by
spectroscopic
analysis
and
multiple-‐step
chemical
derivatiza-‐
tions
(modified
Mosher’s
method,
ozonolysis,
acid
hydrolysis,
Sanger’s
reagent
derivatization,
and
PGME
derivatization).
These
new
lactams
showed
significant
antibacterial
activity
against
gram
negative
bacteria
(Salmonella
enterica,
Proteus
hauseri)
and
cytotoxic
activity
against
several
human
tumor
cell
lines.
Furthermore,
we
analyzed
the
full
genome
of
the
producer
(#SD53)
and
identified
the
biosynthetic
gene
cluster
for
1-‐4.
In
addition,
two
of
new
cyclic
peptides,
UTG9.789
and
UTG9.773
(5-‐6),
were
discovered
from
a
strain
(#UTG9)
belonging
to
the
genus
Microbacterium
isolated
from
burying
beetle‘s
intes-‐
tine.
The
absolute
configurations
of
5-‐6
were
determined
mainly
by
the
advanced
Marfey’s
method
and
GITC
derivatization.
The
structures
of
5-‐6
were
finally
elucidated
as
cyclic
hex-‐
apeptides
bearing
unusual
amino
acids
such
as
chlorinated
tryptophan,
β-‐hydroxy
asparagine,
and
ornithine.
The
major
cyclic
peptide
(5)
showed
antibacterial
activity
against
gram
posi-‐
tive
bacteria
(Staphylococcus
aureus,
Enterococcus
faecalis,
Enterococcus
faecium).
Piceamycin
(1)
and
SD53.449
(2)
Acknowledgements:
This
work
was
supported
by
a
National
Research
Foundation
of
Korea
(NRF)
grant
funded
by
the
Korean
government
(Ministry
of
ICT
and
Future
Planning)
(2014R1A2A1A11053477).
Keywords: Bombyx mori, Nicrophorus concolor, antibacterial, insect-‐associated bacteria,
structure determination, absolute configuration
References:
[1] Crawford
JM,
Clardy
J.
Bacterial
symbionts
and
natural
products.
Chem
Commun
2011;
47:
7559-‐7566
[2] O’Brien
J,
Wright
GD.
An
ecological
perspective
of
microbial
secondary
metabolism.
Curr
Opin
Biotechnol
2011;
22:
552-‐558
[3] Bode
HB.
Insects:
True
pioneers
in
anti-‐infective
therapy
and
what
we
can
learn
from
them.
Angew
Chem
Int
Ed
2009;
48:
6394-‐6396
[4] Schulz
D,
Nachtigall
J,
Riedlinger
J,
Schneider
K,
Poralla
K,
Imhoff
JF,
Beil
W,
Nicholson
G,
Fiedler
H-‐P,
Sussmuth
RD.
Piceamycin
and
its
N-‐acetylcysteine
adduct
is
produced
by
Streptomyces
sp.
GB
4-‐2.
J
Antibiot
2009;
62:
513-‐518
P660
Application
of
a
simple
bioactivity
profiling
strategy
to
natural
product
discovery
from
endophytes
of
marine
macroalgae
Andrew J. Flewelling1, John A. Johnson1, Christopher A. Gray1,2
1 Department of Biological Sciences, University of New Brunswick, 100 Tucker Park Road, E2L 4L5 Saint
John,
Canada,
2
Department
of
Chemistry,
University
of
New
Brunswick,
30
Dineen
Drive,
E3B
5A3
Freder-‐
icton,
Canada
The
natural
products
chemistry
of
marine
macroalgal
endophytes
is
relatively
unexplored
despite
these
fungi
being
recognized
as
a
promising
source
of
new
bioactive
molecules
[1].
As
redundancy
in
natural
products
discovery
increases,
new
techniques
are
needed
to
prioritise
extracts
for
fractionation.
The
use
of
bioactivity
profiling
provides
an
excellent,
albeit
labour
intensive
screening
approach
that
facilitates
the
discovery
of
antibiotics
with
novel
modes
of
action
or
cellular
targets
[2].
Here
we
present
a
simplified
method
for
bioactivity
profiling
that
we
have
applied
to
a
library
of
one
hundred
and
forty-‐one
extracts
of
endophytic
fungi
isolated
from
20
species
of
marine
macroalgae
from
the
Bay
of
Fundy,
Canada.
Extracts
were
screened
for
antimicrobial
activity
against
a
suite
of
Gram
positive
and
Gram
negative
bacte-‐
ria,
mycobacteria
and
fungi.
These
data
were
used
to
compile
bioactivity
profiles
of
each
ex-‐
tract
that
were
compared
to
each
other
and
the
profiles
of
known
antibiotics
representing
a
range
of
modes
of
action.
Principle
component
analysis
revealed
that
34
extracts
exhibited
unique
profiles
within
the
extract
library,
and
hierarchical
cluster
analysis
indicated
six
of
these
extracts
possessed
profiles
different
from
those
of
the
antibiotics.
We
are
currently
sub-‐
jecting
these
six
extracts
to
bioassay-‐guided
fractionation
to
isolate
the
biologically
active
constituents.
We
have
therefore
demonstrated
that
a
simple,
efficient
and
robust
bioactivity
profiling
technique
is
effective
for
prioritising
fungal
extract
libraries.
We
are
confident
that
this
technique
will
be
a
valuable
tool
for
identifying
natural
products
with
unique
antimicro-‐
bial
modes
of
action.
Acknowledgements: Kelsey Pendleton for her assistance with the endophyte isolation
Keywords:
Endophytic
fungi,
bioactivity
profiling,
marine
macroalgae,
antimicrobial,
mode
of
action
References:
[1] Flewelling
AJ,
Currie
J,
Gray
CA,
Johnson
JA.
Endophytes
from
marine
macroalgae:
promis-‐
ing
sources
of
novel
natural
products.
Curr
Sci
2015;
109:
88−111
[2] Wong
WR,
Oliver
AG,
Linington
RG.
Development
of
antibiotic
activity
profile
screening
for
the
classification
and
discovery
of
natural
product
antibiotics.
Chem
Biol
2012;
19:
1483−1495
P661
Rearranged
sesquiterpenes
produced
by
Camarops
sp.
an
endo-‐
phytic
fungus
in
Alibertia
macrophylla
(Rubiaceae)
Juliana
R.
Gubiani1,
Cláudio
R.
Nogueira1,
Maria
C.
M.
Young2,
Paulo
M.
P.
Ferreira3,
Manoel
O.
de
Moraes4,
Cláudia
Pessoa4,
Vanderlan
S.
Bolzani1,
Angela
R.
Araujoa
1
Nucleus
of
Bioassays,
Biosynthesis
and
Ecophysiology
of
Natural
Products
(NuBBE],
Institute
of
Chemis-‐
try,
São
Paulo
State
University
(UNESP),
P.
O.
Box
355,
14800-‐900,
Araraquara,
São
Paulo,
Brazil,
2
Sec-‐
tion
of
Physiology
and
Biochemistry
of
plants,
Botany
Institute,
01061-‐970,
São
Paulo,
Brazil,
3
Experi-‐
mental
Oncology
Laboratory,
Department
of
Biophysics
and
Physiology,
Federal
University
of
Piauí,
64049-‐550,
Teresina,
Piauí,
Brazil.
Sesquiterpenes
are
secondary
metabolites,
which
are
produced
by
a
number
of
endophytic
fungi
[1].
Two
unrelated
families
of
sesquiterpenoids
are
the
botryanes
and
presilphiper-‐
folanes.
Botrydial
displays
phytotoxic,
antibiotic
and
potent
cytotoxic
activities
against
tu-‐
moral
and
non-‐tumoral
cells.
Presilphiperfolanes
shows
antifeedant
activitiy
against
the
chrysomelid
Leptinotarsa
decemlineata
and
the
aphid
Diuraphis
noxia
[1-‐4].
In
our
continual
search
for
bioactive
compounds
from
endophytic
fungi
we
have
investigated
Camarops
sp.
isolated
from
A.
macrophylla
(Rubiaceae).
Camarops
sp
was
cultivated
in
40
500
mL
Erlen-‐
meyer
flasks,
each
containing
90
g
of
corn
and
80
mL
of
H2O.
After
sterilization,
the
medium
was
inoculated
with
the
endophyte
and
incubated
while
stationary
at
25
°C
for
21
days.
The
cultures
were
combined,
ground
and
extracted
with
CH3OH
(7
x
350
mL).
The
solvent
was
evaporated,
yielding
a
crude
CH3OH
extract
(148.3
g),
which
was
dissolved
in
CH3CN
and
defatted
with
hexane.
The
CH3CN
fraction
was
evaporated
to
yield
17.2
g
of
crude
extract,
which
was
fractionated
by
C18
CC
and
eluted
with
an
H2O–CH3OH
gradient
(70:30→0:100)
and
CH3OH–EtOAc
(50:50→0:100),
affording
eight
fractions
(Fr1–Fr8).
Fraction
Fr3
(1,50
g)
and
Fr5
(1.72
g)
after
successive
fractionations
by
CC
and
HPLC
afforded
two
new
compounds
1
(2,5
mg),
2
(7,8
mg)
and
two
known
3
(52.3
mg),
4
(67,2
mg),
respectively.
The
structures
were
determined
as
3,5,9-‐trihydroxy
presilphiperfolane
(1),
4-‐methylenic-‐10-‐
oxodihydrobotrydial
(2),
Xylarenone
C
(3)
and
Xylarenone
D
(4)
by
NMR
and
HRESIMS
analy-‐
sis,
while
the
relative
stereochemistry
of
1
and
2
were
determined
by
NOESY.
The
compounds
3
and
4
exhibited
cytotoxic
activity
against
leukemia
(HL-‐60),
melanoma
(MDA/MB-‐435),
colon
(HCT-‐8)
and
glioblastoma
(SF-‐295)
human
tumor
cell
lines
with
IC50
1.5,
2.4,
1.9,
2.1
and
1.2,
2.1,
1.5,
1.9
µg/mL,
respectively.
Doxorubicin
was
used
as
positive
control.
Compound
4
presented
acetylcholinesterase
inhibition
with
IC50
6.25µg/mL.
Acknowledgements:
We
gratefully
acknowledge
financial
support
from
FAPESP
(Fundação
de
Amparo
`a
Pesquisa
do
Estado
de
São
Paulo,
grant
#2013/07600-‐3)
and
CNPq
(Conselho
Nacional
de
Desenvolvi-‐
mento
Científico
e
Tecnológico)
Grant
#485181/2011.
JRG
also
thanks
CAPES
(Coordenação
de
Aper-‐
feiçoamento
de
Pessoal
de
Nível
Superior)
for
fellowship.
Keywords: Camarops sp., endophytic fungi, sesquiterpenes botrydial, presilfiperfolanes
References:
[1] Collado
IG,
Sánchez
AJM,
Hanson
JR.
Fungal
terpene
metabolites:
biosynthetic
relation-‐
ships
and
the
control
of
the
phytopathogenic
fungus
Botrytis
cinerea.
Nat
Prod
Rep
2007;
24:
674–686
[2] Daoubi
M,
Durán-‐Patrón
R,
Hernández-‐Galán
R,
Benharref
A,
Hanson
JR,
Collado
IG.
The
role
of
botrydienediol
in
the
biodegradation
of
the
sesquiterpenoid
phytotoxin
botrydial
by
Botrytis
cinerea.
Tetrahedron
2006;
62:
8256–8261
[3] Reino
JL,
Durán-‐Patrón
R,
Segura
I,
Hernández-‐Galán
R,
Riese
HH,
Collado
IG.
Chemical
Transformations
on
Botryane
Skeleton.
Effect
on
the
Cytotoxic
Activity.
J
Nat
Prod
2003;
66:
344–349
[4] Durán-‐Patrón
R,
Colmenares
AJ,
Hernández-‐Galán
R,
Collado
IG.
Some
key
metabolic
inter-‐
mediates
in
the
biosynthesis
of
botrydial
and
related
compounds.
Tetrahedron
2001;
57:
1929–1933
P662
Search
for
novel
metabolites
in
fungal
endophytes:
study
of
Pho-‐
mopsis
sp.
and
Colletotrichum
sp.
co-‐cultivation
and
Botry-‐
osphaeria
mamane
epigenetic
modification
Asih
Triastuti1,
Marieke
Vansteelandt1,
Fatima
Barakat1,
Patricia
Jargeat2,
Laura
Rieusset1,
Nicolas
Fabre1,
Carlos
Amasifuen3,
Alexis
Valentin1,
Mohamed
Haddad1
1UMR
152
Pharma
Dev,
Université
de
Toulouse,
IRD,
UPS,
France.
2Université
Paul
Sabatier,
CNRS,
ENFA,
UMR5174
EDB
(Laboratoire
Evolution
et
Diversité
Biologique),
118
route
de
Nar-‐
bonne,
F-‐31062
Toulouse,
France
3Université
Montpellier
2,
UMR
AMAP,
Montpellier,
France;
CNRS,
UMR
AMAP,
Montpellier,
France
Fungal
endophytes
are
fungi
that
grow
within
plant
tissues
without
causing
immediate
signs
of
disease.
They
are
gaining
increased
attention
for
their
ability
to
produce
natural
products
with
diverse
structure
and
pharmacological
activity
[1].
Under
conventional
cultures
condi-‐
tions
some
endophytes
show
loss
of
activity
in
producing
secondary
metabolites
(SM)
due
to
inactivation
of
metabolite
gene
clusters
[2].
Several
strategies
have
been
applied
to
activate
these
silent
gene
clusters
including
epigenetic
modifications
and
co-‐cultivation.
The
signal
come
to
the
cell
will
modify
the
silence
gene
and
lead
to
an
enhanced
production
and/or
to
an
accumulation
of
different
compounds
that
are
not
detected
in
axenic
cultures
[3].
The
purpose
of
this
study
was
to
find
new
SM
and
analyze
gene
clusters
activation
through
co-‐
cultivation
and
through
the
addition
of
molecular
and
epigenetic
modifier
such
as
suberoylan-‐
ilide
hydroxamic
acid
or
valproic
acid
into
the
culture
medium.
Our
research
group
isolated
409
Ascomycetes
strains
from
21
medicinal
plants
from
South
America
[4].
Among
them,
we
chose
Phomopsis
sp.
and
Colletotrichum
sp
on
the
basis
of
ecological
data
as
model
for
co-‐
cultivation
and
Botrysosphaeria
mamane
for
the
epigenetic
modification
studies.
First
results
obtained
highlighted
that
the
fungal
co-‐cultivation
can
induce
de
novo
synthesis
of
secondary
metabolites
(Barakat
et
al.,
data
not
published).
In
order
to
understand
the
mechanisms
un-‐
derlying
these
modifications,
we
focused
on
B.
mamane,
known
to
produce
SM
such
as
mel-‐
lein,
primin,
botryomaman,
and
4,5-‐dihydroxy-‐2-‐hexenoic
acid[5].
This
strain
is
being
under
investigation
in
our
group
for
producing
anti-‐leishmania
SM.
It
exhibited
good
activity
against
Leishmania
infantum
with
IC50=17.
04
µg/ml.
Investigations
are
currently
in
process
to
apply
epigenetic
modifier
in
order
to
induce
SM
production
in
this
strain
and
analyze
its
metabo-‐
lomics
and
pharmacological
activity
modifications
against
L.
infantum
through
immunomodu-‐
latory
approaches.
Keywords:
Fungal
endophytes,
Co-‐cultivation,
Epigenetic,
New
Metabolite
Production
References:
[1] Higginbotham
SJ,
Arnold
AE,
Ibańez
A,
Spadafora
C,
Coley
PD,
Kursar
TA.
Bioactivity
of
fun-‐
gal
endophytes
as
a
function
of
endophyte
taxonomy
and
the
taxonomy
and
distribution
of
their
host
plants.
PLoS
ONE
2013;
8:
e73192
[2] Abdelmohsen
UR,
Grkovic
T,
Balasubramanian
S,
Kamel
MS,
Quinn
RJ,
Hentschel
U.
Elicita-‐
tion
of
secondary
metabolism
in
actinomycetes.
Biotechnol
Adv.
2015;
33:
798-‐811
[3] Marmann
A,
Aly
AH,
Lin
W,
Wang
B,
Proksch
P.
Co-‐cultivation
-‐
a
powerful
emerging
tool
for
enhancing
the
chemical
diversity
of
microorganisms.
Mar
Drugs
2014;
12:
1043-‐1065
[4] Vansteelandt
M,
Jargeat
P,
Haddad
M,
Marti
G,
Fabre
N.
Inside
the
medicinal
plants:
the
fungal
endophytes,
a
hidden
community.
CBS-‐KNAW
Second
Ascomycete
Workshop.
2015
[5] Pongcharoen
W,
Rukachaisirikul
V,
Phongpaichit
S,
Sakayaroj
J.
A
new
dihydrobenzofuran
derivative
from
the
endophytic
fungus
Botryosphaeria
mamane
PSU-‐M76.
Chem
Pharm
Bull
(Tokyo).
2007;
55:
1404-‐1405
P663
Differential
chemotypes
produce
phenotypic
responses
in
endo-‐
phytic
microbial
interactions
Caraballo-‐Rodriguez,
A.
M.,
Pupo,
M.
T.
School
of
Pharmaceutical
Sciences
of
Ribeirao
Preto,
University
of
Sao
Paulo,
Avenida
do
Cafe
s/n,
14040-‐
903
Ribeirao
Preto,
SP,
Brazil
Endophytes
inhabit
plant
tissues
and
share
their
host
with
several
microorganisms
[1].
Re-‐
cently,
we
have
investigated
the
interaction
chemistry
of
Lychnophora
ericoides-‐derived
mi-‐
croorganisms.
We
were
able
to
reveal
a
differential
chemical
repertoire
from
two
endophytic
Actinobacteria,
Streptomyces
albospinus
RLe7
and
S.
mobaraensis
RLe3.
When
challenged
with
an
endophytic
fungus,
Coniochaeta
sp.
FLe4,
a
red-‐pigmentation
in
the
fungal
rim
was
ob-‐
served.
Using
orthogonal
mass
spectrometry
tools
[2,
3],
both
Actinobacteria
showed
distinct
chemical
profiles.
S.
albospinus
RLe7
was
identified
as
an
amphotericin-‐producer
and
it
was
further
demonstrated
that
amphotericin
B
induces
the
phenotypic
‘red-‐rim’
response
in
Coni-‐
ochaeta
sp.
FLe4.
Interestingly,
amphotericin-‐related
compounds
were
not
detected
from
S.
mobaraensis
RLe3,
suggesting
that
different
chemical
entities
are
involved
in
this
fungal
re-‐
sponse.
By
studying
the
chemical
profile
of
S.
mobaraensis
RLe3
we
were
able
to
confirm
the
presence
of
C-‐glycosylated
benz[α]anthraquinones
through
isolation
and
elucidation
of
dehy-‐
droxyaquayamycin
by
NMR
and
HR-‐MS
data,
as
well
as
isolation
and
characterization
of
3
additional
analogues.
Amongst
the
endophytic
fungi
from
L.
ericoides
tested,
only
Coniochaeta
sp.
FLe4
displayed
a
phenotypic
response
in
presence
of
two
different
Actinobacteria
as
well
as
when
chemically
complemented
with
the
pure
natural
products.
In
this
study
it
was
demonstrated
that
individual
chemical
entities
are
responsible
for
specific
chemical
respons-‐
es
(red-‐rim)
in
endophytic
microorganisms
during
interactions.
These
results
may
be
helpful
in
elucidating
chemical
mechanisms
of
microbial
communities
that
occur
inside
the
plant
host.
Cocultures
of
RLe3
and
RLe7
against
FLe4
showing
a
red-‐pigmented
phenotype
RLe3 FLe4 RLe3 FLe4 RLe3 FLe4 RLe7 FLe4 RLe7 FLe4 RLe7 FLe4
Dehydroxyaquayamycin Amphotericin
B
Acknowledgements:
This
work
was
supported
by
grants#2012/21803-‐1
and
2014/01651-‐8
São
Paulo
Research
Foundation
(FAPESP),
Conselho
Nacional
de
Desenvolvimento
Científico
e
Tecnológico
(CNPq)
and
Dorrestein
Lab
at
University
of
California
–
San
Diego
(UCSD).
Authors
also
thank
the
CEPID-‐CIBFar
for
supporting
research
Keywords: endophytes, Lychnophora ericoides, molecular networking, co-‐culture, microbial
interactions.
References:
[1]
Newman
DJ,
Cragg
GM.
Endophytic
and
epiphytic
microbes
as
“sources”
of
bioactive
agents.
Front
Chem
2015;
3:
1−13
[2]
Watrous
J,
Roach
P,
Alexandrov
T,
Heath
BS,
Yang
JY,
Kersten
RD,
van
der
Voort
M,
Pogliano
K,
Gross
H,
Raaijmakers
JM,
Moore
BS,
Laskin
J,
Bandeira
N,
Dorresteina
PC.
Mass
spectral
molecular
networking
of
living
microbial
colonies.
Proc
Natl
Acad
Sci
USA
2012;
109:
E1743−E1752
[3]
Yang
JY,
Phelan
VV,
Simkovsky
R,
Watrous
JD,
Trial
RM,
Fleming
TC,
Wenter
R,
Moore
BS,
Golden
SS,
Pogliano
K,
Dorrestein
PC.
Primer
on
agar-‐based
microbial
imaging
mass
spectrometry.
J
Bacteriol
2012;
194:
6023−6028
P664
Effect
of
germination
on
in
vitro
and
in
vivo
activity
of
Lupinus
al-‐
bus
L.
and
Lupinus
angustifolius
L.
seed
extract
Corina
Danciu1,
Ersilia
Alexa2,
Istvan
Zupko3,
Stefana
Avram1,
Ioana
Zinuca
Pavel1,
Daliana
Minda1,
Georgeta
Pop4,
Dorina
Coricovac5,
Cristina
Dehelean5
1Department
of
Pharmacognosy,
University
of
Medicine
and
Pharmacy
“Victor
Babeş“,
Eftimie
Murgu
Square,
No.
2,
300041
Timişoara,
România,
2 Department
of
Food
Control,
Banat’s
University
of
Agricul-‐
tural
Sciences
and
Veterinary
Medicine
“King
Michael
I
of
Romania”from
Timisoara,
Calea
Aradului
no.119,
300641,
Timisoara,
România,
3
Department
of
Pharmacodynamics
and
Biopharmacy,
University
of
Szeged,
Eötvös
u.
6.,
Szeged
H-‐6720,
Hungary,
4
Department
of
Plant
culture,Banat’s
University
of
Agri-‐
cultural
Sciences
and
Veterinary
Medicine
“King
Michael
I
of
Romania”
from
Timisoara,
Calea
Aradului
no.119,
300641,
Timisoara,
România,
5
Department
of
Toxicology,
University
of
Medicine
and
Pharmacy
“Victor
Babeş“,
Eftimie
Murgu
Square,
No.
2,
300041
Timişoara,
România.
Nutraceuticals
have
lately
been
in
the
spotlight
of
research
due
to
their
significant
contribu-‐
tion
to
the
natural
prevention
of
various
health
issues
[1].
Lupinus
albus
L.,
commonly
known
as
the
white
lupin,
and
Lupinus
angustifolius
L.,
commonly
known
as
the
narrow-‐leafed
lupin
or
blue
lupin,
both
included
in
the
Fabaceae
family,
are
two
plant
species
known
as
nutraceu-‐
ticals
for
their
seeds.
The
Lupinus
genus
comprises
over
200
species
that
contain
approxi-‐
mately
36-‐52%
proteins,
30-‐40%,
fibers
and
5-‐20%
oil
depending
on
the
environmental
or
genetic
conditions
[2,
3].
Ethanolic
extracts
from
the
ungerminated
and
germinated
seeds
of
Lupinus
albus
L.
and
Lupinus
angustifolius
L.
were
analyzed
in
terms
of
the
content
in
total
phenols,
isoflavones
and
cinnamic
acid
derivatives.
Additionally,
the
extracts
were
evaluated
for
antimicrobial
(against
5
bacterial
strains:
Staphylococcus
aureus
(ATCC
25923),
Pseudo-‐
monas
aeruginosa
(ATCC
27853),
Escherichia
coli
(ATCC
25922),
Klebsiella
pneumoniae
(ATCC
700603),
Staphylococcus
epidermidis
(ATCC
14990)),
antiproliferative
(four
cancer
cell
lines:
MCF7,
MDA-‐MB-‐231
(breast
cancers),
A2780
(ovarian
cancer)
and
SiHa
(cervical
cancer))
and
antiinflammatory
properties
(ear
model
of
inflammation),
using
in
vitro
and
in
vivo
tests.
Re-‐
sults
have
shown
that
germination
is
a
method
of
choice
in
increasing
the
amount
of
total
pol-‐
yphenols,
isoflavones
and
cinnamic
acid
derivatives
in
both
Lupinus
albus
L.
and
Lupinus
an-‐
gustifolius
L.
seeds.
However,
biological
evaluation
of
the
antimicrobial,
antiproliferative,
anti-‐
inflammatory
activity
of
all
vegetal
extracts
revealed
an
overall
weak
potential
for
both
un-‐
germinated
and
germinated
seeds.
Keywords: Lupinus albus, Lupinus angustifolius, polyphenols, antibacterial, antiproliferative,
anti-‐inflammatory
References:
[1] Rahal
AM.
Phytonutrients
and
nutraceuticals
in
vegetables
and
their
multi-‐dimensional
medicinal
and
health
benefits
for
humans
and
their
companion
animals:
A
review.
J
Biol
Sci
2014;
14:
1-‐19
[2] Akritidu
KP,
Boinik
VV,
Demeshko
OV.
Organic
acids
from
Lupinus
polyphyllus
roots.
Chem
Nat
Compd
2013;
49:
501-‐502
[3] Mohamed
AA,
Rayas-‐Duarte
P.
Composition
of
Lupinus
albus.
Cereal
Chem
1995;
72:
643-‐
647
P665
Co-‐cultivation
approach
and
untargeted
metabolomics
in
the
search
for
new
secondary
metabolites
from
endophytic
fungi
Fatima
Barakat1,
Marieke
Vansteelandt1,
Asih
Triastuti1,
Laura
Rieusset1,
Billy
Cabanillas2,
Mohamed
Haddad1,
Nicolas
Fabre1
1UMR152
Pharma
Dev,
Université
de
Toulouse,
IRD,
UPS,
France,
2Instituto
de
Investigaciones
de
la
Ama-‐
zonía Peruana (IIAP), Iquitos, Peru
Fungal
endophytes
are
fungi
living
within
the
internal
tissues
of
a
host-‐plant,
without
causing
any
symptoms.
Such
microorganisms
develop
interactions
not
only
with
their
host-‐plant
but
also
with
the
entire
microbiome
of
the
host
[1].
These
organisms,
and
especially
those
isolated
from
plants
of
high
biodiversity
areas,
are
arousing
researchers
interest
since
they
represent
a
relatively
untapped
renewable
source
of
novel
secondary
metabolites
[2].
Nevertheless,
as
micromycetes
can
modulate
their
metabolome
very
fast,
depending
on
their
environment,
some
endophytes
lose
their
ability
to
produce
compounds
of
interest
in
standard
laboratory
conditions
[3].
Thus,
different
strategies
have
been
investigated
to
induce
the
production
of
these
metabolites,
via
the
activation
of
silent
genes.
Such
methods
include
the
addition
of
epi-‐
genetic
modifier
or
a
modification
of
nutrients
into
the
culture
medium,
but
also
the
co-‐
cultivation
of
endophytes.
This
last
approach
may
induce
the
activation
of
some
metabolic
pathway
leading
to
the
production
of
bioactive
metabolites,
in
response
to
the
interactions
between
the
species
[4].
In
order
to
search
for
new
bioactive
compounds
from
fungal
endo-‐
phytes,
we
investigated
two
fungal
strains
isolated
from
a
South
American
plant,
and
belong-‐
ing
to
the
Phomopsis
and
Colletotrichum
genera.
These
strains
were
both
cultured
in
axenic
conditions
and
as
co-‐cultures.
Ethyl
acetate
extractions
of
the
cultures
were
performed
and
extracts
were
analysed
by
Ultra
high-‐performance-‐liquid-‐chromatography
coupled
to
high
resolution
mass
spectrometry
(UHPLC-‐HRMS).
Chemical
fingerprints
obtained
by
untargeted
metabolomics
showed
that
the
co-‐cultivation
modified
the
metabolism
of
the
strains
and
in-‐
duced
the
production
of
new
secondary
metabolites.
In
vitro
bioactivity
of
these
extracts
against
Plasmodium
falciparum,
Leishmania
infantum
and
VERO
cells
is
under
investigation,
such
as
the
isolation
and
characterization
of
the
co-‐cultivation
induced
compounds.
Keywords: fungal endophytes, co-‐cultivation, metabolome, UHPLC-‐HRMS.
References:
[1] Petrini
O.
Fungal
endophytes
of
tree
leaves.
In:
Andrews
JH
and
Hirano
SS,
eds.
Microbial
ecology
of
leaves
1991;
Springer,
Germany.
pp.
179–197.
[2] Strobel
G,
Daisy
B.
Bioprospecting
for
microbial
endophytes
and
their
natural
products.
Microbiol
Mol
Biol
R
2003;
67:
491–502.
[3] Bertrand
S,
Schumpp
O,
Bohni
N,
Bujard
A,
Azzollini
A,
Monod
M,
Wolfender,
J.L.
Detection
of
metabolite
induction
in
fungal
co-‐cultures
on
solid
media
by
high-‐throughput
differential
ultra-‐high
pressure
liquid
chromatography-‐time-‐of-‐flight
mass
spectrometry
fingerprint-‐
ing.
J
Chromatogr
A
2013;
1292:
219–28.
[4] Bertrand
S.,
Bohni
N,
Schnee
S,
Schumpp
O,
Gindro
K
,
&
WolfenderJ.
L.
Metabolite
induc-‐
tion
via
microorganism
co-‐culture:
A
potential
way
to
enhance
chemical
diversity
for
drug
discovery.
Biotechnol
Adv
2014;
32:
1180–1204.
P666
Proteolytic
active
proteins
from
Euphorbia
mauritanica
L.
stimu-‐
late
the
production
of
interleukine-‐6
and
interleukine-‐8
in
keratinocytic
HaCaT
cells
Florian
Guenther,
Matthias
F.
Melzig
Freie
Universitaet
Berlin,
Institute
of
Pharmacy
-‐
Pharmaceutical
Biology,
Königin-‐Luise-‐Str.
2+4,
D-‐
14195
Berlin,
Germany
HaCaT
(human
adult
calcium
low
high
temperature
keratinocytes)
is
a
permanent,
epithelial
human
cell
line,
generated
by
Fusenig
and
Boukamp
[1,2].
Serine
proteases
like
salmon
and
king
crab
trypsin
stimulate
the
interleukine-‐8
production
via
protease-‐activated-‐receptor
2
(PAR2)
[3].
It
is
reported,
that
latices
from
Euphorbiaceae
JUSS.
induce
irritation
and
inflam-‐
mation
on
human
skin.
Proteins
of
Euphorbia
mauritanica
L.
were
isolated
from
latices
by
methanol
precipitation.
After
detection
of
proteolytic
activity,
the
cells
were
treated
with
the
proteolytic
active
frac-‐
tion,
Trypsin,
Phorbol-‐12-‐myristat-‐13-‐acetat
(PMA)
or
the
PAR
agonist
peptide
SFLLR-‐NH2
for
48
hours.
The
supernatant
was
isolated
and
cytokine
concentration
measured
by
enzyme-‐
linked
immunosorbent
assay
(ELISA).
The
quotient
of
cytokine
concentration
and
fluores-‐
cence
of
dsDNA
detection
shows
specific
cytokine
production.
The
results
indicate
that
IL-‐6-‐
release
is
reduced
by
proteins
of
E.
mauritanica
and
IL-‐8
release
is
increased
by
proteins
of
E.
mauritanica
in
combination
with
PMA.
Keywords: Euphorbia mauritanica, proteins, HaCaT, proteases, cytokines
References:
[1] Ryle
CM
et
al.
Density-‐dependent
modulation
of
synthesis
of
keratins
1
and
10
in
the
hu-‐
man
keratinocyte
line
HaCaT
and
in
ras-‐transfected
tumorigenic
clones.
Differentiation
1989;
40:
42-‐54
[2] Boukamp
P
et
al.
Normal
keratinization
in
a
spontaneously
immortalized
aneuploid
hu-‐
man
keratinocyte
cell
line.
J
Cell
Biol
1988;
106:
761-‐771
[3] Bhagwat
SS
et
al.
Salmon
and
king
crab
trypsin
stimulate
interleukin-‐8
and
matrix
metal-‐
loproteinases
via
protease-‐activated
receptor-‐2
in
the
skin
keratinocytic
HaCaT
cell
line.
Food
Chem
Toxicol
2014;
69:
303-‐311
P667
Antigenotoxic
compounds
from
bark
of
South
African
Erythrina
latissima
Yancho
Zarev1,
Kenn
Foubert1,
Vera
Almeida1,
Sandra
Apers1,
Luc
Verschaeve2,
Luc
Pieters1
1
Natural
products
&
Food
Research
and
Analysis
(NatuRA),
Department
of
Pharmaceutical
Sciences,
University
of
Antwerp,
Universiteitsplein
1,
2610
Antwerp,
Belgium,
2
Laboratory
of
Toxicology,
Scientific
Institute
of
Public
Health,
Juliette
Wytsmanstraat
14,
1050
Brussels
Since
mycotoxins
cause
many
diseases,
the
addition
of
protective
plant
substances
to
feeds
and
foods
as
a
chemo-‐preventive
measure
against
the
genotoxic/carcinogenic
effect
of
myco-‐
toxins
seems
a
realistic
approach
[1].
In
preliminary
investigations
bark
extracts
of
Erythrina
latissima
E.Mey
(Leguminosae)
had
shown
antigenotoxic
properties.
Therefore,
the
aim
of
this
work
was
to
isolate
and
to
identify
its
antigenotoxic
constituents.
Dried
and
pulverized
stem
bark
of
E.
latissima
was
percolated
with
dichloromethane/methanol
(1:1).
Subsequent
liquid-‐
liquid
extraction
with
dichloromethane
and
acidified
water,
hexane
and
methanol
(90%)
yielded
5
fractions
(H2O
fraction,
insoluble
fraction
1,
hexane
fraction,
insoluble
fraction
2,
methanol
90%
fraction),
which
were
tested
in
the
Vitotox
test
against
aflatoxin
B1
(AFB1)-‐
induced
mutagenicity
[2].
Apart
from
the
H2O
fraction
all
fractions
displayed
antimutagenic
activity.
Fractions
obtained
were
subjected
to
open
column
chromatography,
normal
and
re-‐
versed
phase
flash
chromatography
and
automated
semi-‐preparative
HPLC-‐MS.
Obtained
compounds
were
elucidated
by
1D
and
2D
NMR
spectroscopy
and
mass
spectrometry.
Several
known
compounds
were
isolated
of
which
the
vast
majority
were
flavanoids,
i.e.
sigmoidin
A,
sigmoidin
B,
sigmoidin
C,
sigmoidin
F,
sigmoidin
G,
4-‐O-‐methylsigmoidin,
abyssinin
II,
abys-‐
sinin
III,
abyssinoflavone
IV,
abyssinoflavone
V,
7-‐demethylrobustegenin,
5’-‐(3methylbut-‐2-‐
enyl)-‐pratensein-‐A,
5,7-‐dihydroxychromone,
5,7,4’-‐trihydroxy-‐3’-‐methoxyflavanone,
Olibergin
A,
Glycyrrhinoflavone.
Also
two
new
compounds
were
identified,
i.e.,
5,7-‐dihydroxy-‐
2-‐[4-‐hydroxy-‐2-‐(prop-‐1-‐en-‐2-‐yl)-‐2,3-‐dihydro-‐1-‐benzofuran-‐5-‐yl]-‐2,3-‐dihydro-‐4H-‐chromen-‐4-‐
one
(1)
and
6,8-‐dihydroxy-‐3-‐(8-‐hydroxy-‐2,2-‐dimethyl-‐3,4-‐dihydro-‐2H-‐chromen-‐6-‐yl)-‐3,4-‐
dihydronaphthalen-‐1(2H)-‐one
(2)
(Fig.
1).
All
major
flavonoids
showed
antigenotoxic
effects
(4-‐100
µg/mL)
without
being
genotoxic.
Mild
cytotoxicity
was
observed
at
100
µg/mL.
The
active
compounds
will
be
quantified
in
the
original
extract
and
the
extract
will
be
evaluated
in
vivo
in
rat
for
its
protective
effect
against
aflatoxin-‐induced
toxicity.
A B OH
O O
HO O HO O
OH
OH O OH O
Figure
.1.
:
chemical
structure
of
compound
1
(A)
and
compound
2
(B).
Acknowledgements:
This
research
was
supported
by
a
research
grant
offered
by
the
Research
Fund
–
Flanders
(FWO),
which
is
also
acknowledged
for
granting
a
post-‐doctoral
fellowship
to
KF
Keywords: Erythrina latissima, flavonoid, antigenotoxicity, vitotox assay
References:
[1] Turner
NW,
Subrahmanyam
S,
Piletsky
SA.
Analytical
methods
for
determination
of
myco-‐
toxins:
A
review.
Anal
Chem
Acta
2009;
632:
168-‐180
[2] Verschaeve
L,
Van
Gompel
J,
Thilemans
L,
Regniers
L,
Vanparys
P,
van
der
Lelie
D.
VITO-‐
TOX¨
Bacterial
Genotoxicity
and
Toxicity
Test
for
the
Rapid
Screening
of
Chemicals.
Envi-‐
ron
Mol
Mutagen
1999;
33:
240-‐248
P668
Chemical
constituents
and
biological
activity
from
the
stem
and
root
of
Neolitsea
konishii
Hsien-‐Kai
Huang1,
Shan-‐Yu
Lin2,
Su-‐Ling
Wong3,
Tian-‐Lu
Cheng4,
Chu-‐Hung
Lin1,
Kim-‐Hong
Gan1,
Hsun-‐Shuo
Chang1,2,3,
Ih-‐Sheng
Chen1,2,3
1
School
of
Pharmacy,
College
of
Pharmacy,
Kaohsiung
Medical
University,
Kaohsiung
807,
Taiwan,
2
Graduate
Institute
of
Natural
Products,
College
of
Pharmacy,
Kaohsiung
Medical
University,
Kaohsiung
807,
Taiwan,
3
Research
Center
for
Natural
Products
and
Drug
Development,
Kaohsiung
Medical
Univer-‐
sity,
Kaohsiung
807,
Taiwan,
4
Department
of
Biomedical
Science
and
Environmental
Biology,
Kaohsiung
Medical
University,
Kaohsiung
807,
Taiwan
Sixty-‐three
species
of
Formosan
indigenous
lauraceous
plants
were
screened
under
the
drug-‐
screening
platforms
of
Escherichia
coli
β-‐glucuronidase
(eβG)
inhibitors
against
chemothera-‐
py-‐induced
diarrhea
(CID).
The
methanolic
extract
of
the
stem
and
the
root
of
Neolitsea
kon-‐
ishii
(Hayata)
Kanehira
&
Sasaki
can
specifically
block
intestinal
eβG-‐activity
but
not
affect
the
human
βG
(hβG),
respectively.
N.
konishii
is
a
small
evergreen
tree,
distributes
in
Ryukyu
and
Taiwan
[1].
In
Taiwan,
it
grows
in
low
to
medium
altitudes
forest.
Sesquiterpenoids,
mono-‐
terpenoids,
triterpenoids,
alkaloids,
benzenoids,
and
steroids
have
previously
been
reported
from
the
bark,
leaves,
fruits
of
N.
konishii
[2].
Therefore,
the
aim
of
this
study
was
to
isolate
the
chemical
constituents
of
this
species
and
to
evaluate
their
anti-‐eβg
activity.
Bioassay-‐
guided
fractionation
of
the
active
ethyl
acetate-‐soluble
layer
of
the
stem
of
N.
konishii
led
to
the
isolation
of
one
new
1,3-‐diphenylbutanoid,
(‒)-‐konishibutanin,
two
new
lignans,
neok-‐
oninins
A
and
B,
and
two
new
β-carboline alkaloids, neolitseacarbolines A and B,
along
with
20
known
compounds.
One
new
alkanone
derivative,
neolitsenol,
along
with
24
known
com-‐
pounds
were
isolated
from
the
root
of
N.
konishii.
The
structures
of
these
isolates
were
eluci-‐
dated
by
spectral
analysis.
Among
the
isolates,
132-hydroxy-(132-R)-pheophytin a and N-trans-
feruloyl-3ʹ,4ʹ-dihydroxyphenylethylamine
can
specifically
block
intestinal
eβG-‐activity
(inhibi-‐
tion
ratio
of
88%
and
76%)
but
not
affect
the
hβG.
Some
analog
of
N-‐trans-‐feruloyl-‐3ʹ,4ʹ-‐
dihydroxyphenylethylamine
were
also
evaluated
on
anti-‐eβg
activity
but
no
one
showed
sig-‐
nificant
activity.
Acknowledgements:
This
work
was
partially
supported
by
grants
from
the
Kaohsiung
Medical
University
“Aim
for
the
Top
Universities
Grant,
grant
No.
KMU-‐TP103H01
and
KMU-‐TP103H05”
Keywords: Neolitsea konishii, Lauraceae, root, stem, anti-‐Escherichia coli β-‐glucuronidase
References:
[1] Li
HL.
Lauraceae.
In:
Flora
of
Taiwan,
Vol.
2,
2nd
edition.
Taipei,
Taiwan:
Editorial
Com-‐
mittee
of
the
Flora
of
Taiwan;
1996:
433−499
[2] Chang
HS,
Chen
IS.
Chemical
constituents
and
bioactivity
of
Formosan
lauraceous
plants.
J
Food
Drug
Anal
2016,
24:
247−263
P669
Cytotoxic
lignans
from
the
fruits
of
Koelreuteria
henryi
Chu-‐Hung
Lin1,
Kuo-‐Hsiung
Lee2,
Ih-‐Sheng
Chen1,3,4,
Hsun-‐Shuo
Chang1,3,4,5
1
School
of
Pharmacy,
College
of
Pharmacy,
Kaohsiung
Medical
University, Kaoshiung
807,
Taiwan,
2
Nat-‐
ural
Products
Research
Laboratories,
UNC
Eshelman
School
of
Pharmacy,
University
of
North
Carolina,
Chapel
Hill,
North
Carolina
27599,USA,
3
Graduate
Institute
of
Natural
Products,
College
of
Pharmacy,
Kaohsiung
Medical
University,
Kaoshiung
807,
Taiwan,
4Research
Center
for
Natural
Products
and
Drug
Development,
Kaohsiung
Medical
University,
Kaoshiung
807,
Taiwan,
5
Center
for
Infectious
Disease
and
Cancer
Research
(CICAR),
Kaohsiung
Medical
University,
Kaohsiung
807,
Taiwan
Koelreuteria
henryi
Dummer
(Sapindaceae)
is
a
large
evergreen
tree,
endemic
to
Taiwan
[1].
The
methanolic
extract
of
the
fruits
of
K.
henryi
showed
strong
cytotoxicity
against
MCF-‐7
(Breast
cancer
cells),
A549
(Lung
cancer),
KB
(Epidermoid
carcinoma
of
the
nasopharynx),
KB-‐VIN
(Vincristine
resistant),
and
MDA-‐MB-‐231
(Triple-‐negative
breast
cancer)
cancer
cell
lines
in
vitro.
The
aim
of
this
study
is
the
isolation
of
chemical
constituents
from
this
plant
and
the
evaluation
of
their
biological
activities.
Previously,
we
reported
13
compounds,
including
three
new
lignans
and
one
isolated
from
nature
for
the
first
time,
along
with
nine
known
compounds
from
the
active
ethyl
acetate-‐soluable
layer
of
this
plant.
Continuing
investigation
of
the
active
layer
of
this
species
led
to
the
isolation
of
one
new
lignan
derivative,
namely
keolreuterone
E
(5).
The
structures
of
these
new
compounds
(1‒3,
and
5)
were
elucidated
by
1D,
2D
NMR,
UV,
IR,
ESIMS,
and
HRESIMS
analysis.
In
this
study,
compounds
1-‐3,
6,
7,
and
8
were
evaluated
for
cytotoxic
activities
against
the
MCF-‐7,
A549,
KB,
KB-‐VIN
and
MDA-‐MB-‐231
cancer
cell
lines
and
paclitaxel
was
used
as
positive
control.
Among
these
isolates,
keolreuter-‐
one
C
(3)
and
austrobailignan-‐1
(6)
displayed
potent
cytotoxicity
against
the
KB,
KB-‐VIN,
and
MCF-‐7
cancer
cell
lines,
with
IC50
values
of
5.800,
5.484,
and
9.147;
0.045,
0.007,
and
0.074
μM,
respectively.
However,
the
structure
of
austrobailignan-‐1
(6)
was
similar
to
podophyllo-‐
toxin and showed the most cytotoxic activity [2].
Acknowledgements:
This
work
was
supported
by
grants
from
the
National
Science
Council
of
Republic
of
China,
“NSC
99-‐2320-‐B-‐037-‐010-‐MY3”,
and
partially
supported
by
grants
from
the
Kaohsiung
Medical
University
“Aim
for
the
Top
Universities
Grant,
grant
No.
KMU-‐TP103H01,
KMU-‐TP103H05
and
KMU-‐
TP104E43”
Keywords: Koelreuteria henryi, Sapindaceae, fruits, lignan, cytotoxic activity
References:
[1] Chen
CH.
Sapindaceae.
In:
Flora
of
Taiwan,
Vol.
3,
2nd
edition.
Taipei,
Taiwan:
Editorial
Committee
of
the
Flora
of
Taiwan;
1993:
599−608
[2] Song
YN,
Zhang
HL,
Chang
CJ.
Cytotoxic
cyclolignans
from
Koelreuteria
henryi.
J
Nat
Prod
1994,
57:
1670−1674
P670
Three
new
dikeopiperazines
from
Costa
Rican
endolichenic
fun-‐
gus
Colpoma
sp.
CR1465A
Jae
Sik
Yu1,
Seulah
Lee1,
Hee
Jeong
Eom1,
Hee
Rae
Kang1,
Seoung
Rak
Lee1,
Tae
Kyoung
Lee1,
Jiwon
Baek1,
Dahae
Lee1,
Won
Se
Suh1,
Ki
Hyun
Kim1
1School
of
Pharmacy,
Sungkyunkwan
University,
Suwon
440-‐746,
Republic
of
Korea
Three new diketopiperazines (1–3), namely cyclo(L-‐Pro-‐D-‐trans-‐Hyp) (1), cyclo(L-‐Pro-‐D-‐Glu)
(2), and cyclo(D-‐Pro-‐D-‐Glu) (3) and five known diketopiperazines (4–8) were isolated from
the endolichenic fungus Colpoma sp. CR1465A identified from the Costa Rican plant Henri-‐
ettea tuberculosa (Melatomataceae). The structures of the new compounds 1–3 were eluci-‐
dated using a combination of extensive spectroscopic analyses, including 2D NMR and HR-‐MS,
and their absolute configurations were determined by a combination of NOESY analysis and
Marfey’s method [1,2].Cyclo(L-‐Pro-‐D-‐allo-‐Thr) (4) was recently isolated from a South China
Sea marine sponge Callyspongia sp. [3], but its NMR spectroscopic data were not reported,
and cyclo(L-‐Pro-‐L-‐Asp) (5) was previously reported but only as a synthetic product [4]. The
NMR data assignments of compounds 4 and 5 are reported for the first time. All of the isolated
compounds were tested for antifungal and antimicrobial properties.
O O O O O
7 H H H OH O
HO H
12
6
1 N HO 9 N 12
HO N 11
3 8 8
N
N 4 HN HN
9 HN
H 10 H 10 H
O O H 9
O O
1 2 3 4
O O O O
H
O
8 N 11 N N N
OH HN HN HN HN
H 9 H H H
O O O O
5 6 7 8
Acknowledgements:
This
paper
was
supported
by
SEOK
CHUN
Research
Fund,
Sungkyunkwan
University,
2014
Keywords: Colpoma sp., Henriettea tuberculosa, Diketopiperazines, Marfey’s method
References:
[1] Noh
HJ,
Hwang
D,
Lee
ES,
Hyun
JW,
Yi
PH,
Kim
GS,
Lee
SE,
Pang
C,
Park
YJ,
Chung
KH,
Kim
GD,
Kim
KH.
Anti-‐inflammatory
activity
of
a
new
cyclic
peptide,
citrusin
XI,
isolated
from
the
fruits
of
Citrus
unshiu.
J
Ethnopharmacol
2015;
163:
106-‐112
[2] Marfey
P.
Determination
of
D-‐amino
acids.
II.
Use
of
a
bifunctional
reagent,
1,5-‐difluoro-‐
2,4-‐dinitrobenzene.
Carlsberg
Res
Commun
1984;
49:
591-‐596
[3] Chen
Y,
Peng
Y,
Gao
C,
Huang
R.
A
new
diketopiperazine
from
South
China
Sea
marine
sponge
Callyspongia
sp.
Nat
Prod
Res
2014;
28:
1010-‐1014
[4] Challa
C,
Kumar
N,
John
M,
Lankalapalli
RS.
A
comparative
study
of
antimicrobial
proper-‐
ties
of
cyclo(L -‐Pro-‐
L -‐Asp)
with
its
2-‐ketopiperazine
analog.
Med
Chem
Res
2014;
23:
2377-‐2385
P671
Resorcylic
lactones
from
Lasiodiplodia
theobromae
(MUB65),
a
fungal
endophyte
isolated
from
Myracrodruon
urundeuva
Aline
C.
M.
Sobreira1,3,
Otília
D.
L.
Pessoa1,
Katharine
G.
D.
Florêncio2,
Diego
V.
Wilke2,
Francis-‐
co
C.
O.
Freire3,
Francisco
J.
T.
Gonçalves3,
Paulo
Riceli.
V.
Ribeiro3,
Lorena
M.
A.
Silva3,
Edy.
S.
Brito3,
Kirley
M.
Canuto3.
1Departamento
de
Química
Orgânica
e
Inorgânica,
Universidade
Federal
do
Ceará,
Campus
do
Pici,
60021-‐97,
Fortaleza,
Brazil,
2Núcleo
de
Pesquisa
e
Desenvolvimento
de
Medicamentos,
Universidade
Fed-‐
eral
do
Ceará,
Rua
Coronel
Nunes
de
Melo,
1000,
60430-‐275
Fortaleza,
Brazil,
3Embrapa
Agroindústria
Tropical,
Rua
Dr.
Sara
Mesquita,
2270,
60511-‐110
Fortaleza,
Brazil
Endophytic
fungi
have
been
pointed
out
as
a
promising
source
of
pharmacologically
active
substances,
among
compounds
with
potential
anticancer
action
[1].
Here
in,
we
carried
out
a
chemical
investigation
and
cytotoxicity
tests
of
a
Lasiodiplodia
theobromae
strain
(MUB-‐65)
isolated
from
the
branches
of
Myracrodruon
urundeuva
(Anacardiaceae),
which
is
a
native
tree
from
Northeastern
Brazil
used
as
an
anti-‐inflammatory
medicine.
The
fungus
was
grown
in
a
malt
medium
for
21
days
and
afterwards,
its
broth
was
partitioned
with
ethyl
acetate.
The
EtOAc
extract
was
analyzed
by
ultra-‐performance
liquid
chromatography
with
high-‐
resolution
mass
spectrometry
(UPLC-‐HRMS).
Sephadex
LH-‐20
fractionation
and
high-‐
performance
liquid
chromatography
of
this
extract
yielded
two
yellowish
solids
named
(1)
and
(2)
(Fig.
1).
UPLC-‐HRMS
chromatogram
showed
15
peaks
that
were
characterized
as
two
isocoumarins,
two
lasiojasmonates,
one
eremophilane
sesquiterpene,
two
lasiodiplodin
de-‐
rivatives
as
well
as
1
and
2,
besides
six
unknown
compounds.
The
structures
of
(1)
and
(2)
were
elucidated
by
1D
and
2D
nuclear
magnetic
resonance.
The
compound
1
was
identified
as
lasiodiplodin,
a
macrolide
reported
earlier
in
L.
theobromae
from
other
host-‐plants
and
in
plants
[3,4].
The
compound
(2)
was
determined
as
rel-‐11-‐12-‐(7'R*,
4'R*,
2'R*-‐
tetrahydrofuro[1',2']pyranyl)-‐lasiodiplodin,
which
is
being
described
in
the
literature
for
the
first
time.
The
cytotoxicity
of
these
compounds
was
tested
against
the
colorectal
cancer
cell
line
HCT-‐116,
using
the
MTT
in
vitro
assay
[2].
Compound
(1)
exhibited
moderate
effect,
pre-‐
senting
an
IC50
of
11.2
μg.mL-‐1,
while
(2)
showed
no
activity.
In
previous
studies,
lasiodiplodin
was
cytotoxic
against
an
ovarian
cancer
cell
line
and
demonstrated
potent
antileukemic
effect
in
a
lymphocytic
leukemia
in
vivo
assay
[3,4].
Furthermore,
resorcylic
lactones
are
well-‐
known
as
protein
kinase
inhibitors,
which
are
currently
one
of
the
main
mechanism
of
action
of
commercial
anticancer
drugs
[5].
OCH3 O
OCH3 O
O
O
O
HO
O
H
H
1
2
Acknowledgements:
Thanks
to
the
Banco
do
Nordeste
do
Brasil
for
the
financial
support
(Convênio
7385).
Keywords: endophytic fungus, lasiodiplodin, macrolide, citotoxicity.
References:
[1] Chandra
S.
Endophytic
fungi:
novel
sources
of
anticancer
lead
molecules.
Appl
Microbiol
Biotechnol
2012;
95:
47–59
[2] Mosmann
T.
Rapid
colorimetric
assay
for
cellular
growth
and
survival:
application
to
pro-‐
liferation
and
cytotoxicity
assays.
J
Immunol
Methods
1983;
65:
55–63
[3] Lee
K-‐H,
Hayashi
N,
Okamo
M,
Hall
IH,
Wu
R-‐Y,
McPhailti
AT.
Lasiodiplodin,
a
potent
antileukemic
macrolide
from
Euphorbia
splendens.
Phytochemistry
1982;
21:
1119–1121
[4] Cao
S,
Hou,
Y,
Brodie
P,
Miller
JS,
Randrianaivo
R,
Rakotobe
E,
Rasamison
VE,
Kingston,
DG.
Antiproliferative
compounds
of
Cyphostemma
greveana
from
Madagascar
dry
florest.
Chemistry
&
Biodiversity
2011;
8:
643–650
[5] Patocka
J,
Soukup
O,
Kuca
K.
Resorcylic
acid
lactones
as
the
protein
kinase
inhibitors,
nat-‐
urally
occuring
toxins.
Mini-‐Rev
Med
Chem
2013;
13:
1873–1878
P672
Biodiversity
of
fungal
community
associated
to
lichens
active
against
Candida
biofilms
Patricia
Jargeat1,
Marion
Girardot2,
Caroline
Rouger3,
Willy
Aucher2,
Marion
Millot3,
Christine
Imbert2,
Lengo
Mambu3
1
UMR5174
UPS-‐CNRS-‐ENFA
Laboratory
of
Evolution
and
Biological
Diversity,
University
of
Toulouse,
Bât
4R1,
118
route
de
Narbonne,
31062
Toulouse,
France;
2
UMR
CNRS
7267
Laboratory
of
Ecology
and
Biol-‐
ogy
of
Interactions,
Faculty
of
Medicine
Pharmacy,
University
of
Poitiers,
Bât
D1,
6
rue
de
la
Milétrie,
TSA
51115,
86073
Poitiers
cedex
9,
France;
3
EA
1069
Laboratory
of
Chemistry
of
Natural
Substances,
Faculty
of
Pharmacy,
University
of
Limoges,
2
rue
du
Dr
Marcland,
87025
Limoges
cedex,
France
Lichens
are
symbiotic
organisms
and
their
thalli
constitute
an
ecological
niche
for
associated
microbial
communities
(fungi
and
bacteria)
classified
as
epi-‐
and
endobionts,
possibly
able
to
form
biofilm-‐like
structures
[1].
Interactions
exist
within
this
complex
ecosystem
and
appear
to
be
at
the
origin
of
the
production
of
defense
metabolites
[2]
which
contribute
to
the
re-‐
sistance
of
lichens
to
fungi.
These
compounds
also
display
interesting
biological
activities
such
as
antimicrobials
[3]
or
inhibitors
of
bacterial
biofilms
[4].
A
screening
of
fifty
lichen
ex-‐
tracts
was
previously
performed
against
sessile
Candida
albicans
yeasts
by
evaluating
their
anti-‐adherent
activity
and
highlighted
the
interest
of
7
lichen
extracts.
In
order
to
explore
the
possible
link
between
fungal
inhibition
of
these
extracts
and
biotic
existing
balances
within
the
lichen,
the
study
of
fungal
community
associated
to
thalli
of
active
lichens
was
performed.
Endolichenic
and
epilichenic
fungi
were
isolated
on
MEA
and
PDA
culture
media
after
sterili-‐
sation
step
or
not
of
the
thalli
respectively
[5]
and
the
characterisation
of
the
total
fungal
community
by
high-‐throughput
sequencing
(MiSeq)
from
crushed
thalli
was
carried
out.
Se-‐
quencing
of
the
ITS
rDNA
and/or
18S
and
comparison
with
sequence
databases
were
per-‐
formed
from
cultures.
The
results,
especially
with
thalli
of
the
2
most
active
lichens
(Evernia
prunastri
and
Ramalina
fastigiata)
showed
a
wide
fungal
diversity.
The
endolichenic
versus
epilichenic
communities
are
well
differentiated
with
a
predominance
of
Sordariomycetes
for
endolichenic
fungi
and
Dothideomycetes
for
epilichenic
fungi.
Eurotiomycetes,
Leotiomycetes
and
Pezizomycetes
are
also
present.
Epi
and
endobionts
belong
to
classes
that
are
distinct
from
that
of
mycobiont
(Lecanoromycetes)
[5].
Only
the
species
Sordaria
fimicola
was
found
to
be
both
epi
and
endolichenic
fungus.
The
analyse
of
results
obtained
by
high-‐throughput
sequencing
after
ITS1
amplification
is
in
progress.
Acknowledgements:
CNRS
is
acknowledged
for
financial
support
(Licafilm
project
/
Exomod
PEPS
re-‐
search
program
2014,
2015)
Keywords: Lichens, epilichenic and endolichenic fungi, Candida albicans biofilm, Ascomycetes
References:
[1] Grube
M,
Berg
G.
Microbial
consortia
of
bacteria
and
fungi
with
focus
on
the
lichen
symbi-‐
osis.
Fungal
biol
rev
2009;
23:
72-‐85
[2] Lawrey
JD,
Rossman
AY,
Lowen
R.
Inhibition
of
selected
hypocrealean
fungi
by
lichen
sec-‐
ondary
metabolites.
Mycologia
1994;
86:
502-‐506
[3] Zambare
VP,
Christopher
LP.
Biopharmaceutical
potential
of
lichens.
Pharm
biol
2012;
50:
778-‐798
[4] Francolini
I,
Norris
P,
Piozzi
A,
Donelli
G,
Stoodley
P.
Usnic
acid,
a
natural
antimicrobial
agent
able
to
inhibit
bacterial
biofilm
formation
on
polymer
surfaces.
Antimicrob
Agents
Chemother
2004;
48:
4360-‐4365
[5] Arnold
AE,
Miadlikowska
J,
Higgins
KL,
Sarvate
SD,
Gugger
P,
Way
A,
Hofstetter
V,
Kauff
F,
Lutzoni
F.
A
phylogenetic
estimation
of
trophic
transition
networks
for
ascomycetous
fungi:
are
lichens
cradles
of
symbiotrophic
fungal
diversification?
Syst
Biol
2009;
58:
283-‐
297
P673
Increasing
the
production
of
secondary
metabolites
by
changes
in
culture
conditions
Madelinea
Aguilar1,2,
Maria
Julca-‐Canto1,
Nivia
Rios3,
Luis
Cubilla-‐Rios1,2
1Laboratory
of
Tropical
Bioorganic
Chemistry,
Faculty
of
Natural,
Exact
Sciences
and
Technology,
Apartado
0824-‐00159,
University
of
Panama,
Panama
City,
Panama,
2Smithsonian
Tropical
Research
Institute,
Panama
City,
Panama,
3Department
of
Microbiology,
Faculty
of
Natural,
Exact
Sciences
and
Technology,
Apartado
0824-‐00159,
University
of
Panama,
Panama
City,
Panama.
In
our
search
for
active
compounds
from
macro
and
micro
fungi,
a
number
of
active
molecules
have
been
isolated
from
the
cultures.
Such
is
the
case
of
3-‐epi-‐waol
A
(1)
[1],
mangiferaelac-‐
tone
(2)[2]
and
mycoleptodiscin
B
(R=OH,
3)[3],
which
have
shown
biological
activity
(cyto-‐
toxic
and
antibacterial).
Thereafter,
four
research
groups
have
reported
the
synthesis
of
man-‐
giferaelactone
[4];
while
the
synthesis
of
mycoleptodiscin
A
(R=H,
4)
[5],
an
analog
of
myco-‐
leptodiscin
B,
has
also
been
reported
once.
An
alternative
way
to
obtain
these
active
compounds,
which
avoids
the
use
of
complicated
strategies
of
synthesis
as
well
as
the
use
of
chemicals
that
could
be
adverse
to
the
environ-‐
ment,
is
the
exploration
of
new
methodologies
to
culture
the
microorganism.
Recently,
we
have
cultivated
the
saprophytic
fungus
Lentinus
striguelus
[6]
following
different
strategies
and
as
a
result,
a
group
of
new
secondary
metabolites
has
been
isolated.
Moreover
we
have
increased
the
production
of
panepoxydone
(5),
an
epoxyketone
produced
by
some
species
in
the
genus
Lentinus,
for
which
previous
studies
had
determined
its
antiparasitic
and
anticancer
activity
[7,8],
Could
the
development
of
biotechnological
strategies
be
an
alternative
for
the
production
of
interesting
secondary
metabolites
produced
by
fungi?
O
NH
OH O OH
O
O
H
OH
O
O O O
O R
2 5
HO
O R=OH, 3
H
1 O OH R=H, 4 HO
Acknowledgements:
For
financial
support
we
thank
to
The
Global
Environment
Facility
(GEF),
The
Na-‐
tional
Secretary
of
Science,
Technology
and
Innovation
(SENACYT-‐Panamá)
and
The
International
Coop-‐
erative
Biodiversity
Group
(ICBG-‐Panamá).
Keywords: Fungi, panepoxydone, magiferaelactone, mycoleptodiscin B.
References:
[1] Adames
I,
Ortega
HE,
Asai
Y,
Kato
M,
TenDyke
K,
Shen
YY,
Cubilla-‐Ríos
L.
3-‐epi-‐Waol
A
and
Waol
C:
polyketide-‐derived
γ-‐lactones
isolated
from
the
endophytic
fungus
Libertella
blepharis
F2644.
Tetrahedron
Lett
2015;
56:
252-‐255
[2] Ortega
HE,
TenDyke
K,
Shen
YY,
Rios
N,
Cubilla-‐Ríos
L.
An
Antibacterial
Polyhydroxylated
Macrolide
Isolated
from
the
endophytic
fungus
Pestalotiopsis
mangiferae.
Tetrahedron
Lett
2014;
55:
2642-‐2645
[3] Ortega
HE,
Graupner
PR,
Asai
Y,
Kato
M,
TenDyke
K,
Qiu
D,
Shen
YY,
Rios
N,
Arnold
AE,
Coley
PD,
Kursar
TA,
Gerwick
WH,
Cubilla-‐Ríos
L.
Mycoleptodiscins
A
and
B,
cytotoxic
al-‐
kaloids
from
the
endophytic
fungus
Mycoleptodiscus
sp.
F0194.
J
Nat
Prod
2013;
76:
741-‐
744
[4] Reddy
BVS,
Reddy
PS,
Babu
KV,
Reddy
BP,
Yadav
JS.
Stereoselective
Total
Synthesis
of
Mangiferaelactone
using
d-‐Mannose
as
a
Chiral
Pool.
Helv
Chim
Acta
2015;
98:
1395-‐
1402
[5] Zhou
S,
Chen
H,
Luo
Y,
Zhang
W,
Li
A.
Asymmetric
total
synthesis
of
Mycoleptodiscin
A.
Angew
Chem
Int
Edit
2015;
54:
6878–6882
[6] Julca-‐Canto
M,
Rios
N,
Aguilar
M,
Sousa
JPB,
Cubilla-‐Ríos
L.
Additional
new
natural
prod-‐
ucts
produced
by
Lentinus
strigellus:
A
biotechnological
approach.
Tetrahedron
Lett
2016;
57:
650-‐653.
[7] Barros-‐Filho
BA,
Oliveira
MCF,
Mafezoli
J,
Barbosa
FG,
Rodrigues-‐Filho
E.
Secondary
me-‐
tabolite
production
by
the
Basidiomycete,
Lentinus
strigellus,
under
different
culture
con-‐
ditions.
Nat
Prod
Commun
2012;
7:
771-‐773
[8] Arora
R,
Schmitt
D,
Karanam
B,
Tan
M,
Yates
C,
Dean-‐Colomb
W.
Inhibition
of
the
Warburg
effect
with
a
natural
compound
reveals
a
novel
measurement
for
determining
the
meta-‐
static
potential
of
breast
cancers.
Oncotarget
2015;
6:
662-‐678
P674
New
hormonemate
derivatives
from
the
endophytic
fungus
Dothiora
sp.
Mercedes
Pérez-‐Bonilla1,
Víctor
González-‐Menendez1,
Nuria
de
Pedro1,
Ignacio
Pérez-‐
Victoria1,
Jesús
Martín1,
Francisca
Vicente1,
Olga
Genilloud1,
José
R.
Tormo1,
Fernando
Reyes1
1Fundación
MEDINA,
Centro
de
Excelencia
en
Investigación
de
Medicamentos
Innovadores
en
Andalucía,
Parque Tecnológico Ciencias de la Salud. Avda. del Conocimiento 34, 18016, Armilla, Granada, Spain
Arid
zones
in
Andalucía
have
special
edaphological
and
climatic
conditions
such
as
low
rain-‐
fall,
high
sunshine
levels
and
specific
lithology
where
loamy
materials
and
evaporites
abound.
Native
plant
communities
from
these
arid
zones
possess
distinctive
survival
characteristics
in
these
special
conditions,
which
have
led
to
the
existence
of
a
high
degree
of
endemic
plants
with
highly
adapted
endophyte
ecosystems
poorly
studied.
It
is
precisely
this
singularity
which
turns
them
into
a
valuable
potential
source
for
the
isolation
of
new
unique
host-‐specific
endophytes.
Fungal
endophytes
are
known
to
produce
a
wide
variety
of
secondary
metabo-‐
lites
involved
in
their
adaptation
and
survival
within
higher
plants.
Launaea
arborescens
was
collected
in
Tabernas
desert
(Andalucía,
Spain),
an
arid
area
of
the
Almería
province,
being
one
of
the
representative
plants
characteristic
of
this
geographic
re-‐
gion.
The
fungal
strain
Dothiora
sp.
was
isolated
from
this
plant
and
the
taxonomic
determina-‐
tion
of
its
genus
and
species
was
performed
by
sequencing
the
complete
ITS1-‐5.8S-‐ITS2-‐28S
region
or
independent
ITS
and
28S
rDNA
compared
with
GenBank
and
the
NITE
Biological
Resource
Center
databases
by
using
the
BLAST
application
[1].
Acetone
extract
from
the
strain
grown
in
the
YES
medium
showed
cytotoxicity
against
2
cancer
cell
lines:
human
breast
adenocarcinoma
(MCF-‐7)
and
liver
carninoma
(HEPG2).
Bioassay-‐guided
fractionation
of
this
extract
using
SP207ss
resin
column
chromatography
and
preparative
reversed-‐phased
HPLC
led
to
the
isolation
of
six
new
natural
products,
hormonemates
B-‐G,
structurally
related
to
hormonemate
[2]
whose
chemical
characterization
is
reported
herein
for
the
first
time.
Cyto-‐
toxic
activities
of
the
pure
compounds
against
human
liver
carcinoma
(HEPG2),
breast
(MCF-‐
7)
and
pancreatic
carcinoma
(MiaPaca)
were
determined.
Moderate
cytotoxicity
in
the
mi-‐
cromolar
range
was
found
for
four
of
these
compounds
against
the
HEPG2
(IC50
=
26-‐36
µM)
and
MCF-‐7
(IC50
=
11-‐28
µM)
cell
lines.
Acknowledgements:
To
the
Junta
de
Andalucía
for
financial
support
through
the
Project
RNM-‐7987.
Keywords: Launaea arborescens, fungal endophytes, Dothiora sp., hormonemate, cytotoxicity.
References:
[1] González-‐Menéndez
V,
Pérez-‐Bonilla
M,
Pérez-‐Victoria
I,
Martín
J,
Muñoz
F,
Reyes
F,
Tormo
JR,
Genilloud
O.
Multicomponent
Analysis
of
the
Differential
Induction
of
Secondary
Me-‐
tabolite
Profiles
in
Fungal
Endophytes.
Molecules
2016;
21:
234–250
[2] Filip
P,
Weber
RWS,
Sterner
O,
Anke
T.
Hormonemate,
a
New
Cytotoxic
and
Apoptosis-‐
Inducing
Compound
from
the
Endophytic
Fungus
Hormonema
dematioides.
I.
Identifica-‐
tion
of
the
Producing
Strain,
and
Isolation
and
Biological
Properties
of
Hormonemate.
Z
Naturforsch
2003;
58:
547–552
P675
Angiogenesis
inhibitors
and
anti-‐inflammatory
agents
from
Pho-‐
ma
sp.
NTOU4195
Ming-‐Shain
Lee1,
Shin-‐Wei
Wang2,
Guei-‐Jane
Wang3,
Ka-‐Lai
Pang4,
Ching-‐Kuo
Lee5,
Yueh-‐
Hsiung
Kuo6,
Hyo-‐Jung
Cha7,
Ruo-‐Kai
Lin8,
Tzong-‐Huei
Lee9
1School
of
Pharmacy,
Taipei
Medical
University,
Taipei,
Taiwan
11031,
2Department
of
Medicine,
Mackay
Medical
College,
New
Taipei
City,
Taiwan
25245,
3Graduate
Institute
of
Clinical
Medical
Science,
China
Medical
University,
Taichung,
Taiwan
40402,
4Institute
of
Marine
Biology
and
Center
of
Excellence
for
the
Oceans,
National
Taiwan
Ocean
University,
Keelung,
Taiwan
20224,
5Graduate
Institute
of
Pharmacy,
Taipei
Medical
University,
Taipei,
Taiwan
11031,
6Department
of
Chinese
Pharmaceutical
Sciences
and
Chinese
Medicine
Resources,
China
Medical
University,
Taichung,
Taiwan
40447,
7Institute
of
Marine
Bi-‐
ology
and
Center
of
Excellence
for
the
Oceans,
National
Taiwan
Ocean
University,
Keelung,
Taiwan
20224,
8Graduate
Institute
of
Pharmacy,
Taipei
Medical
University,
Taipei,
Taiwan
11031,
9Institute
of
Fisheries
Science,
National
Taiwan
University,
Taipei,
Taiwan
10617.
Seven
new
polyketides,
namely
phomaketides
A‒E
(1-‐5),
pseurotins
A3
(6),
and
G
(7),
along
with
FR-‐111142
[1],
pseurotins
A,
A1,
A2,
D,
F2,
14‒norpseurotin
A
[2,3],
α-‐carbonylcarbene,
tyrosol,
cyclo-‐L-‐Pro-‐L-‐Leu,
and
cyclo-‐L-‐Pro-‐L-‐Phe
were
purified
from
the
fermented
broth
and
mycelium
of
an
endophytic
fungal
strain
Phoma
sp.
NTOU4195
isolated
from
a
marine
red
algae
Pterocladiella
capillacea.
The
chemical
structures
were
established
by
interpretations
of
the
spectroscopic
data.
The
anti-‐angiogenic
and
anti-‐inflammatory
effects
of
the
new
isolates
1‒7
and
their
known
analogues
were
evaluated
by
using
human
endothelial
progenitor
cells
(EPCs)
and
lipopolysaccharide
(LPS)-‐activated
murine
macrophage
RAW264.7
cells,
respec-‐
tively.
Of
these
compounds
tested,
compound
1
exhibited
the
most
potent
anti-‐angiogenic
activity
by
suppressing
tube
formation
of
EPCs
with
an
IC50
value
of
8.1
±
0.5
μM,
while
com-‐
pound
3
showed
the
most
selective
inhibitory
activity
on
LPS-‐induced
NO
production
in
RAW264.7
macrophages
with
an
IC50
value
of
8.8
±
0.3
μM.
Acknowledgments:
This
work
was
supported
by
grants
from
the
Ministry
of
Science
and
Technology.
We
thank
Ms.
S.-‐L.
Huang
of
the
Instrumentation
Center
of
the
College
of
Science,
National
Taiwan
University,
for
the
NMR
data
acquisition
and
Ms.
Y.-‐C.
Wu
of
the
Small
Molecule
Metabolomics
Core
Facility,
the
Insti-‐
tute
of
Plant
and
Microbial
Biology
and
Academia
Sinica
Scientific
Instrument
Center,
Academia
Sinica,
for
the
MS
data
acquisition
Keywords:
Phoma
sp.,
Pterocladiella
capillacea,
polyketide,
endophyte,
ntric
oxide,
anti-‐
angiogenic.
References:
[1] Otsuka
K,
Shibata
T,
Tsurumi
Y,
Takase
S,
Okuhara
M,
Terano
H,
Kohsaka
M,
Imanaka
H.
A
new
angiogenesis
inhibitor,
FR-‐111142.
J
Antibiot
1992;
45:
348-‐354.
[2] Wang
FZ,
Li
DH,
Zhu
TJ,
Zhang
M,
Gu
QQ.
Pseurotin
A1
and
A2,
two
new
1-‐oxa-‐7-‐
azaspiro[4.4]non-‐2-‐ene-‐4,6-‐diones
from
the
holothurian-‐derived
fungus
Aspergillus
fu-‐
migatus
WFZ-‐25.
Can
J
Chem
2011;
89:
72-‐76.
[3] Breitenstein
W,
Chexal
KK,
Mohr
P,
Tamm
C.
Pseurotin
B,
C,
D,
and
E.
Further
new
metabo-‐
lites
of
Pseudeurotium
ovalis
STOLK.
Helv
Chim
Acta
1981;
64:
379-‐388.
P676
Discovery
of
new
bioactive
secondary
metabolites
produced
by
Streptomyces
strains
derived
from
volcanic
islands
Munhyung
Bae1,
Heegyu
Kim2,
Sohyun
Park1,
Jongheon
Shin1,
Ki-‐Bong
Oh2,
Sang
Kook
Lee1,
Dong-‐Chan
Oh1
1
Natural
Products
Research
Institute,
College
of
Pharmacy,
Seoul
National
University,
1
Gwanak-‐ro,
Gwanak-‐gu,
Seoul
08826,
Republic
of
Korea,
Republic
of
Korea,
2
Department
of
Agricultural
Biotechnol-‐
ogy,
College
of
Agriculture
and
Life
Science,
Seoul
National
University,
1
Gwanak-‐ro,
Gwanak-‐gu,
Seoul
08826,
Republic
of
Korea
Microorganisms
have
been
regarded
as
one
of
the
most
efficient
natural
sources
for
drug
dis-‐
covery
and
development
over
the
past
decades
including
the
golden
era
of
antibiotics
during
1960s
[1,
2].
However,
the
decline
of
discovering
new
antibiotics
by
conventional
approaches
and
constant
clinical
need
for
bioactive
compounds
with
structural
novelty
have
led
natural
products
chemists
to
investigate
microorganisms
that
inhabit
relatively
under-‐investigated
environments
[3,
4],
such
as
abandoned
mines,
salterns,
the
Arctic
sea,
and
deep-‐sea.
As
part
of
efforts
to
search
for
new
bioactive
compounds,
we
have
focused
on
actinomycetes
derived
from
climatically
different
two
volcanic
islands
in
Republic
of
Korea,
Jeju
and
Ul-‐leung
Islands.
First,
SAK30
strain
was
isolated
from
a
soil
sample
collected
in
Jeju
Island.
The
planar
struc-‐
ture
of
SAK30A
(1)
was
determined
as
a
new
polyene
peroxide
bearing
hexadeca-‐
2,4,6,8,10,12,14-‐heptaenoic
acid
chain
by
comprehensive
analysis
of
1D
and
2D
NMR
spec-‐
troscopy.
Further
analysis
of
1H-‐1H
coupling
constants
and
ROESY
correlations
resulted
in
the
determination
of
relative
configuration
of
the
1,2-‐oxolane
ring
and
the
double
bond
geome-‐
tries
of
1.
The
planar
structures
of
SAK30B-‐D
(2-‐4)
were
also
elucidated
based
on
the
analysis
of
1D
and
2D
NMR
spectroscopy.
SAK30A
(1),
bearing
the
endoperoxide
moiety,
strongly
dis-‐
played
antimicrobial
activity.
Secondly,
the
chemical
analysis
of
Streptomyces
strain
SUD119,
which
was
isolated
from
a
sediment
sample
collected
in
Ul-‐leung
Island,
indicated
that
SUD119
produced
a
new
benz
[a]anthracene
dimer
linked
by
a
sulfide
bond.
The
planar
struc-‐
ture
of
SUD119A
(5)
was
clarified
based
on
the
mass
data,
UV
spectrum,
and
comprehensive
spectroscopic
analyses.
The
stereochemistry
of
SUD119A
(5)
was
determined
by
the
analysis
of
ROESY
correlations
with
DFT
calculation
and
the
ECD
calculations.
SUD119A
(5)
exhibited
significant
quinone
reductase
induction
activity.
Keywords:
Volcanic
island,
polyene
peroxide,
benz
[a]
anthracene,
antibiotic
References:
[1] Bérdy
J.
Thoughts
and
facts
about
antibiotics:
Where
we
are
now
and
where
we
are
head-‐
ing.
J
Antibiot
2012;
65:
385-‐395
[2] Ortholand
J-‐Y,
Ganesan
A.
Natural
products
and
combinatorial
chemistry:
back
to
the
fu-‐
ture.
Curr
Opin
Chem
Biol
2004;
8:
271-‐280
[3] Fenical
W,
Jensen
PR.
Developing
a
new
resource
for
drug
discovery:
marine
actinomycete
bacteria.
Nat
Chem
Biol
2006;
2:
666-‐673
[4] Pettit
RK.
Culturability
and
secondary
metabolite
diversity
of
extreme
microbes:
expand-‐
ing
contribution
of
deep
sea
and
deep-‐sea
vent
microbes
to
natural
product
discovery.
Mar
Biotechnol
2011;
13:
1-‐11
P677
Biocontrol
potential
of
endophytic
fungi
against
Batrachochytri-‐
um
dendrobatidis,
the
fungi
causing
global
amphibian
declines
Carolina
Castro,
Carolina
Portero,
Alexandra
Narváez-‐Trujillo
Laboratory
of
Plant
Biotechnology,
Department
of
Biological
Sciences,
Pontificia
Universidad
Católica
del
Ecuador,
Quito,
Ecuador
The
pathogenic
fungus
Batrachochytrium
dendrobatidis
(Bd)
causes
the
disease
called
chytridiomycosis
[1]
and
has
contributed
to
the
dramatic
decline
of
frog
species
around
the
world
[2].
The
information
collected
in
the
last
decade
suggests
that
at
least
26
species
of
am-‐
phibians
in
Ecuador
have
suffered
major
declines
in
their
populations
because
of
this
pathol-‐
ogy
[3].
Worldwide
various
species
of
frogs
have
been
affected
[4].
Organic
extracts
from
twenty-‐two
Ecuadorian
endophytes
cryopreserved
at
-‐80°C
from
the
Endophyte
Collection
Quito
Catolica
(CEQCA)
were
evaluated
in
in
vitro
bioassays
for
their
capacity
to
control
or
inhibit
growth
of
Batrachochytrium
dendrobatidis.
Protocols
using
Alamar
Blue®
were
stand-‐
ardized
for
microplate
determination
of
minimum
inhibitory
concentrations
(MICs)
of
6
en-‐
dophytes
with
fungicide
activities
against
Batrachochytrium
dendrobatidis.
Additionally,
6
endophytes
producing
volatile
organic
compounds
were
evaluated.
Four
volatile
organic
compounds
(VOCs)
producing
endophytes
presented
total
inhibition
of
Bd
growth;
qualitative
data
of
this
inhibition
is
presented.
Up
to
date,
conservation
of
amphibian
species
depends
on
rescue
centres
and
reproduction
in
captivity
since
no
natural
in
situ
mechanisms
to
control
Bd
disease
have
been
established.
Rescue
centres
rely
on
commercial
fungicides
[5].
We
propose
the
use
of
natural
products
based
on
endophyte
extracts
and
VOCs
as
alternatives.
Keywords:
Endophytes,
amphibian
decline,
Ecuador,
Batrachochytrium
dendrobatidis
References:
[1] Rosenblum
EB,
Voyles
J,
Poorten
TJ,
Stajich
JE.
The
Deadly
Chytrid
Fungus:
A
Story
of
an
Emerging
Pathogen.
PLoS
Pathog
2010;
6:
1-‐2
[2] Stuart,
S.
N.,
J.
S.
Chanson,
N.
A.
Cox,
B.
E.
Young,
A.
S.
L.
Rodrigues,
D.
L.
Fischman,
R.
W.
Waller.
Status
and
trends
of
amphibian
declines
and
extinctions
worldwide.
Science
2004;
306:1783-‐1786
[3] Ron
S.
R.,
Merino-‐Viteri
A.
Amphibian
declines
in
Ecuador:
overview
and
first
report
of
chytridiomycosis
from
South
America.
Froglog
2000;
42:
2-‐3
[4] Pessier
AP,
Mendelson
JR
(eds).
A
manual
for
control
of
infectious
diseases
in
amphibian
survival
assurance
colonies
and
reintroduction
programs.
Apple
Valley,
MN:
IUCN/SCC
Conservation
Breeding
Specialist
Group
2010
[5] Bennett
M.
Hardy,
Karen
L.
Pope,
Jonah
Piovia-‐Scott,
Richard
N.
Brown,
Janet
E.
Foley.
Itra-‐
conazole
treatment
reduces
Batrachochytrium
dendrobatidis
prevalence
and
increases
over
winter
field
survival
in
juvenile
Cascades
frogs.
Dis
Aquat
Org
2015;
112:
243-‐250
P678
Chemical
study
on
Paenibacillus
odorifer,
a
bacterial
species
iso-‐
lated
from
lichen
Nguyen
Thi
Bach
Le,
Delmail
David,
Tomasi
Sophie
UMR
CNRS
ISCR
6226
PNSCM,
Department
of
Pharmacy
and
Biological
Sciences,
University
of
Rennes
1,
2
avenue
du
Pr.
Léon
Bernard,
35043
Rennes,
France.
Recent
studies
have
admitted
that
bacterial
communities
were
an
important
united
partner
of
mini-‐ecosystem
lichen
symbiosis
[1].
Lichen,
moreover,
is
also
a
rich
source
of
new
bacterial
lineages
and
novel
bacterial
compounds
[2].
Therefore,
microorganism
communities
isolated
from
lichens
became
a
significant
subject
as
a
great
potential
of
natural
product
discovery.
Paenibacillus
odorifer,
a
bacterium
belonging
to
Firmicutes,
was
isolated
from
Rhizocarpon
geographicum,
a
particular
popular
crust-‐forming
rock
lichen
[3].
Assessing
its
optimal
growth
conditions
was
conducted
to
improve
its
metabolite
production
yield
by
measure
of
optical
density
(OD)
and
by
analysis
of
its
chemical
profile
via
TLC,
HPLC,
LC-‐MS.
Firstly,
the
parameters
chosen
for
optimal
culture
were
kind
of
medium,
pH
and
temperature.
Results
revealed
that
the
growth
of
P.
odorifer
was
better
at
250C
using
Gym
Streptomyces
liquid
me-‐
dium
supplemented
with
CaCO3
at
pH=7.
Secondly,
the
fermentation
was
launched
by
apply-‐
ing
the
best
conditions
of
bacterial
growth.
After
the
3-‐day
culture
using
a
bioreactor,
the
su-‐
pernatant
and
bacterial
cells
were
separated
by
centrifugation
and
organic
compounds
were
absorbed
from
supernatant
by
XAD-‐7
resin.
The
desorption
from
resin
was
carried
out
through
several
stages
to
collect
raw
extract.
TLC,
HPLC
and
LC-‐MS
analyses
highlighted
that
this
extract
was
a
rich
source
of
metabolites.
Then,
the
crude
extract
was
fractionated
via
flash
chromatography
on
a
40
g
C18
reversed-‐phase
column,
using
a
gradient
solvent
system
of
CH3CN-‐H2O
at
15
mL/min
flow
rate
in
75
min.
Two
diketopiperazines
were
isolated
from
the
fractions
by
applying
semi-‐preparative
HPLC
(Prevail
C18
column,
various
ratios
of
CH3CN)
such
as
cyclo(L-‐Phe-‐L-‐Val)
and
cyclo(L-‐Phe-‐L-‐Ile).
Correspondingly,
fermentation
of
P.
odorifer
could
produce
valuable
compounds
for
future
research.
Acknowledgments:
We
acknowledge the
“Ligue
Contre
le
Cancer”
for
financial
support
for
the
BioFlo®
115
bioreactor.
Keywords:
Paenibacillus
odorifer,
Rhizocarpon
geographicum,
diketopiperazine,
HPLC,
LC-‐MS.
References:
[1]
Grube
M,
Cernava
T,
Soh
J,
Fuchs
S,
Aschenbrenner
I,
Lassek
C,
Wegner
U,
Becher
D,
Riedel
K,
Sensen
CW,
Berg
G.
Exploring
functional
contexts
of
symbiotic
sustain
within
Lichen-‐
associated
bacteria
by
comparative
omics.
ISME
J
2015;
2:
412–424
[2]
Suzuki
MT,
Parrot
D,
Berg
G,
Grube
M,
Tomasi
S.
Lichens
as
natural
sources
of
biotechno-‐
logically
relevant
bacteria.
Appl
Microbiol
and
Biotechnol
2016;
2:
583−595
[3]
Bjelland
T,
Grube
M,
Hoem
S,
Jorgensen
SL,
Daae
FL,
Thorseth
IH,
Øvreås
L.
Microbial
meta-‐
communities
in
the
lichen–rock
habitat.
Environ
Microbiol
Rep
2011;
4:
434–442
P679
Bigger
is
not
always
better:
A
study
of
the
structure-‐activity
rela-‐
tionship
of
oligomeric
ellagitannins
on
ruminal
fermentation
in
vitro
Nicolas
Baert1,
Wilbert
F.
Pellikaan2,
Maarit
Karonen1,
Juha-‐Pekka
Salminen1
1
Laboratory
of
Organic
Chemistry
and
Chemical
Biology,
Department
of
Chemistry,
University
of
Turku,
Turku
FI-‐20014,
Finland,
2
Animal
Nutrition
Group,
Department
of
Animal
Sciences,
Wageningen
Univer-‐
sity,
P.O.
Box
338,
NL-‐6700
AH
Wageningen,
The
Netherlands
Methane
from
enteric
fermentation
is
a
natural
by-‐product
of
the
digestive
processes
of
rumi-‐
nants
which
represents
a
loss
of
2
to
12%
of
the
animal’s
gross
energy
intake
[1].
Plant
sec-‐
ondary
metabolites
and
tannins
in
particular,
have
shown
some
promising
results
as
dietary
inhibitors
of
methanogenesis.
However,
the
structure-‐activity
relationships
that
underlie
the
mechanisms
of
action
of
tannins
in
the
rumen
remain
mostly
unkown.
In
this
study,
we
inves-‐
tigated
the
effect
of
the
degree
of
oligomerization
of
ellagitannins
(ET)
on
their
ability
to
alter
ruminal
fermentation.
For
that
purpose
we
isolated
dimeric
to
heptameric
ET
from
rosebay
willowherb
(Epilobium
angustifolium)
and
tested
these
compounds
on
several
indicators
of
ruminal
fermentation
with
an
in
vitro
gas
production
system.
Results
show
that
oligomeric
ET
decreased
total
volatile
fatty
acids
production
proportionally
to
their
degree
of
oligomeriza-‐
tion.
Methane
production
was
also
decreased
but
the
experiment
did
not
reveal
any
difference
in
the
level
of
activity
of
the
ET.
Additionally,
willowherb’s
oligomeric
ET
decreased
ammonia-‐
nitrogen
and
branched-‐chain
volatile
fatty
acids
concentrations,
thus
indicating
reduced
pro-‐
tein
degradation
by
ruminal
bacteria.
In
conclusion
this
study
shows
that
the
various
activi-‐
ties
of
willowherb’s
oligomeric
ET
towards
ruminal
fermentation
are
highly
dependent
on
their
chemical
structure.
But
more
importantly,
smaller
oligomers
had
less
detrimental
effect
toward
organic
matter
fermentation
than
the
larger
ones
while
displaying
similar
level
of
an-‐
timethanogenic
activity.
250 a
ab
Total volatile fatty acids
cd bc
(mmol/L/g OM)
200
de
e
ef
f
150
er
er
s
l
er
er
er
er
tro
er
im
am
am
im
m
m
on
m
ta
tra
Tr
D
ly
ex
nt
C
ep
Te
Po
Pe
Treatment
Keywords:
Gas
production
technique,
willowherb,
methane,
structure-‐activity
relationship
References:
[1] Johnson
KA,
Johnson
DE.
Methane
emissions
from
cattle.
J
Anim
Sci
1995;
73:
2483-‐2492
P680
Elicitation
of
isoflavonoid
production
in
the
cell
suspension
cul-‐
ture
of
Pueraria
candollei
var.
candollei
by
endophytic
bacteria
Panitch
Boonsnongcheep1,
Atsuko
Matsumoto2,3,
Yoko
Takahashi2,3,
Sompop
Prathanturarug1
1Department
of
Pharmaceutical
Botany,
Faculty
of
Pharmacy,
Mahidol
University,
447
Sri-‐Ayuthaya
Road,
Rajathevi,
Bangkok
10400,
Thailand,
2Graduate
School
of
Infection
Control
Sciences,
Kitasato
Uni-‐
versity,
5-‐9-‐1
Shirokane,
Minato-‐ku,
Tokyo
108-‐8641,
Japan,
3Kitasato
Institute
for
Life
Sciences,
Kitasato
University,
5-‐9-‐1
Shirokane,
Minato-‐ku,
Tokyo,
108-‐8641,
Japan
Tuberous
roots
of
Pueraria
candollei
var.
candollei
were
traditionally
used
in
Thailand
as
re-‐
juvenator.
The
cell
suspension
culture
of
P.
candollei
var.
candollei
can
produce
high
amount
of
isoflavonoids,
daidzin
and
genistin,
in
short
time
[1]. Some
elicitors
accelerated
the
accu-‐
mulation
of
isoflavonoids
in
P.
candollei
cell
suspension
cultures
[2].
This
present
study
focus-‐
es
on
the
usage
of
endophytic
bacteria,
isolated
from
P.
candollei,
as
the
elicitors.
The
bacteria
were
isolated
from
roots
and
root
nodules
of
P.
candollei
in
Thailand.
They
were
preliminary
identified
by
the
sequence
of
16S
rRNA
and
EzTaxon
database
[3].
After
identification,
7
rhi-‐
zobia
and
13
actinomycete
isolates
were
selected
and
used
for
the
elicitation
with
cell
suspen-‐
sion
culture
of
P.
candollei
var.
candollei.
The
accumulation
of
isoflavonoids
in
the
cell
suspen-‐
sion
culture
was
analyzed
by
HPLC.
The
addition
of
living
rhizobia
isolates
could
elicit
the
production
of
isoflavonoids
in
the
P.
candollei
var.
candollei
up
to
8.72±0.17
and
mg/g
dry
weight,
higher
than
the
untreated
group
and
to
the
comparable
level
with
yeast
extract
(7.09±0.79
mg/g
dry
weight)
and
methyl
jasmonate
(8.17±1.78
mg/g
dry
weight).
In
the
sim-‐
ilar
manner,
the
50%
ethanol
extract
of
actinomycete
culture
broths
could
enhance
the
isofla-‐
vonoid
production
in
the
cell
suspension
culture,
up
to
6.15±1.16
mg/g
dry
weight,
1.5
times
higher
than
the
control
group.
In
conclusion,
the
results
indicated
the
capability
of
endophytic
rhizobia
and
the
extract
of
endophytic
actinomycetes
as
the
elicitors
for
the
isoflavonoid
pro-‐
duction
in
P.
candollei
var.
candollei
cell
suspension
culture.
Acknowledgements:
This
study
was
supported
by
the
Thailand
Research
Fund
through
the
Royal
Golden
Jubilee
Ph.D.
Program
(Grant
no.
PHD/0302/2551)
and
the
Office
of
the
High
Education
Commission
and
Mahidol
University
under
the
National
Research
Universities
initiative
Keywords: Cell suspension culture, endophytes, elicitation, isoflavonoids
References:
[1] Boonsnongcheep
P,
Korsangruang
S,
Soonthornchareonnon
N,
Chintapakorn
Y,
Saralamp
P,
Prathanturarug
S.
Growth
and
isoflavonoid
accumulation
of
Pueraria
candollei
var.
candollei
and
P.
candollei
var.
mirifica
cell
suspension
cultures.
Plant
Cell
Tiss
Organ
Cult
2010;
101:
119-‐126
[2] Korsangruang
S,
Soonthornchareonnon
N,
Chintapakorn
Y,
Saralamp
P,
Prathanturarug
S.
Effects
of
abiotic
and
biotic
elicitors
on
growth
and
isoflavonoid
accumulation
in
Pueraria
candollei
var.
candollei
and
P.
candollei
var.
mirifica
cell
suspension
cultures.
Plant
Cell
Tiss
Organ
Cult
2010;
103:
333-‐342
[3] Kim
OS,
Cho
YJ,
Lee
K,
Yoon
SH,
Kim
M,
Na
H,
Park
SC,
Jeon
YS,
Lee
JH,
Yi
H,
Won
S,
Chun
J.
Introducing
EzTaxon-‐e:
a
prokaryotic
16S
rRNA
gene
sequence
database
with
phylotypes
that
represent
uncultured
species.
Int
J
Syst
Evol
Microbiol
2012;
62:
716-‐721
P657
A
new
indole
alkaloid
from
the
endophyte
Aspergillus
ustus
iso-‐
lated
from
the
mangrove,
Avicennia
germinans
Petrea
C.
Facey1,
Roy
B.
Porter1,
Hartmut
Laatsch2
1Department
of
Chemistry,
University
of
the
West
Indies,
Mona,
Kingston
17,
Jamaica;
2Department
of
Organic
Chemistry,
University
of
Goettingen,
Tammannstrasse
2,
D-‐37077
Goettingen,
Germany
Endophytic
fungi
are
microorganisms
that
spend
the
whole
or
part
of
their
life
cycle
inside
the
living
tissues
of
their
host
plants
in
a
symbiotic
or
pathogenic
relationship.
These
endophytes
have
been
found
to
produce
a
wide
range
of
biologically
active
compounds
which
may
con-‐
tribute
to
the
protection
and
survival
of
their
host
plant
[1].
Mangroves,
due
to
their
harsh
habitat
of
high
salinity,
high
temperature
and
high
UV
radiation,
offer
a
particularly
unique
environment
for
the
isolation
of
these
endophytes
[2].
As
a
result,
the
mangrove,
Avicennia
germinans
(Black
mangrove)
found
along
the
coastal
waters
of
Jamaica,
was
investigated
for
associated
fungal
endophytes.
The
fungus
Aspersgillus
ustus
was
isolated
and
its
rice
cultures
extracted
and
chromatographed
via
column
chromatography
using
silica
gel
and
Sephadex
LH-‐20.
A
new
prenylated
indole
alkaloid
(1)
and
three
previously
isolated
sesterterpenoids
ophiobolin
G
(2)
[3],
ophiobolin
K
(3a)
and
6-‐epiophiobolin
K
(3b)
[4]
were
obtained
from
the
ethyl
acetate
extract.
Their
structures
were
elucidated
using
1D
and
2D
NMR
and
MS
data
analysis.
O
H
N
N O
H
O
O N
H
1
Acknowledgements:
This
work
was
funded
by
the
Office
of
Sponsored
Research,
University
of
the
West
Indies,
Jamaica
and
the
Humboldt
Foundation,
Germany.
We
are
grateful
to
Dr.
Lynne
Sigler,
University
of
Alberta
Microfungus
Collection
and
Herbarium,
Canada
for
the
identification
of
fungus.
Keywords: Indole alkaloid, Aspergillus ustus, Avicennia germinans, endophytes
References:
[1] Strobel
G,
Daisy
B,
Castillo
U,
Harper
J.
Natural
products
from
endophytic
microorganisms.
J
Nat
Prod
2004;
67:
257−268
[2] Ding
L,
Hans-‐Martin
D,
Hertweck
C.
Cytotoxic
alkaloids
from
Fusarium
incarnatum
associ-‐
ated
with
the
mangrove
tree
Aegiceras
corniculatum.
J
Nat
Prod
2012;
75:
617−621.
[3] Cutler
GH,
Crumley
FG,
Cox
RH,
Springer
JP,
Arrendale
RF,
Cole
RJ,
Cole
PD.
Ophiobo-‐
lins
G
and
H:
new
fungal
metabolites
from
a
novel
source,
Aspergillus
ustus.
J
Agric
Food
Chem
1984;
32:
778−782.
[4] Singh
SB,
Smith
JL,
Sabnis
GS,
Dombrowski
AW,
Schaeffer
JM,
Goetz
MA,
Bills
GF.
Structure
and
conformation
of
ophiobolin
K
and
6-‐epiophiobolin
K
from
Aspergillus
ustus
as
a
nem-‐
atocidal
agent.
Tetrahedron
1991;
47:
6931−693
P682
Compounds
with
anti-‐respiratory
syncytial
virus
activity
from
endophytic
fungus
Pestalotiopsis
thea
Philip
F.
Uzor1,
Damian
C.
Odimegwu2,
Weaam
Ebrahim3,4,
Patience
O.
Osadebe1,
Ngozi
J.
Nwodo1,
Festus
B.
C.
Okoye5,
Zhen
Liu3,
Peter
Proksch3
1
Department
of
Pharmaceutical
and
Medicinal
Chemistry,
University
of
Nigeria,
Nsukka,
410001,
Nigeria,
2
Division
of
Pharmaceutical
Microbiology
and
Biotechnology,
Department
of
Pharmaceutics,
University
of Nigeria, Nsukka, 410001, Nigeria, 3Heinrich-‐Heine-‐Universität, Institut für Pharmazeutische Biologie
und Biotechnologie, Universitatsstrasse 1, Geb. 26.23, Düsseldorf, 40225, Germany, 4 Department of
Pharmacognosy, Faculty of Pharmacy, Mansoura University, Mansoura 35516, Egypt, 5 Department of
Pharmaceutical and Medicinal Chemistry, Faculty of Pharmaceutical Sciences, Nnamdi Azikiwe Universi-‐
ty, Awka, Nigeria
Respiratory syncytial virus (RSV) is a known cause of severe respiratory infections with at-‐
tendant high morbidity and mortality rates in all populations especially in infants younger
than 2 years of age [1,2].Currently, the viral infection has no approved vaccine and the only
approved drug, ribavirin, is expensive and is not likely to improve outcome [3]. This has war-‐
ranted the search for safer and more potent alternatives from natural sources such as medici-‐
nal plants and endophytic fungi. Endophytic fungi have been known as prolific source of bio-‐
active compounds [4]. The present study was aimed at investigating the anti-‐RSV activity of
compounds from endophytic fungi. The endophytic fungus Pestalotiopsis thea was isolated
from the leaves of the host Nigerian plant Fagara zanthoxyloids (Rutaceae). After fermenta-‐
tion in solid rice media for 21 days, the fungus afforded three known compounds (Fig. 1):
chloroisosulochrin (1), ficipyrone A (2) and pestheic acid (3) which were identified from their
spectroscopic and physicochemical data and comparison with the literature [5,6]. The com-‐
pounds were investigated for their anti-‐RSV activity using the HEP-‐2 cell lines and ribavirin as
the standard drug. Compound 1 was found to show a strong inhibition of the RSV with IC50 of
4.22 ± 1.03 µM (ribavirin 4.91 ± 1.85 µM). Other compounds showed moderate inhibition of
the virus (IC50 ranging from 45.00 ± 0.98 to 146.20 ± 2.14 µM). The results of the present
study have shown that chloroisosulochrin (1), isolated from an endophytic fungus P. thea,
possesses strong activity against RSV and could be developed as a potent drug against the vi-‐
ral infection.
Fig.
1:
Isolated
compounds
from
Pestalotiopsis
thea
Keywords:
Chloroisosulochrin,
endophytic
fungi,
ficipyrone
A,
Pestalotiopsis
thea,
pestheic
acid,
respiratory
syncytial
virus.
References:
[1] Hall
CB,
Weinberg
GA,
Iwane
MK,
Blumkin
AK,
Edwards
KM,
Staat
MA,
Auinger
P,
Griffin
MR,
Poehling
KA,
Erdman
D,
Grijalva
CG,
Zhu
Y,
Szilagyi
P.
The
burden
of
respiratory
syn-‐
cytial
virus
infection
in
young
children.
New
Engl
J
Med
2009;
360:
588-‐598
[2] Bloom-‐Feshbach
K,
Alonso
WJ,
Charu
V,
Tamerius
J,
Simonsen
L,
Miller
MA,
Viboud
C.
Latitu-‐
dinal
variations
in
seasonal
activity
of
influenza
and
respiratory
syncytial
virus
(RSV):
a
global
comparative
review.
PLoS
ONE
2013;
8:
Article
ID
e54445
[3] Schickli
JH,
Dubovsky
F,
Tang
RS.
Challenges
in
developing
a
pediatric
RSV
vaccine.
Hum
Vaccin
2009;
5:
582-‐591
[4] Uzor
PF,
Ebrahim
W,
Osadebe
PO,
Nwodo
JN,
Okoye
FB,
Müller
WEG,
Lin
W,
Liu
Z,
Proksch
P.
Metabolites
from
Combretum
dolichopetalum
and
its
associated
endophytic
fungus
Nigrospora
oryzae
–
Evidence
for
a
metabolic
partnership.
Fitoterapia
2015;
105:
147-‐150
[5] Shimada
A,
Takahashi
I,
Kawano
T,
Kimura
Y.
Chloroisosulochrin,
chloroisosulochrin
de-‐
hydrate,
and
pestheic
acid,
plant
growth
regulators,
produced
by
Pestalotiopsis
theae.
Z
Naturforsch
2001;
56b:
797-‐803
[6] Liu
S,
Liu
X,
Guo
L,
Che
Y,
Liu
L.
2H-‐Pyran-‐2-‐one
and
2H-‐furan-‐2-‐one
derivatives
from
the
plant
endophytic
fungus
Pestalotiopsis
fici.
Chem
Biodivers
2013;
10:
2007-‐2013
P683
The
endophyte
Candidatus
Burkholderia
crenata
of
the
TCM
plant
Ardisia
crenata
produces
the
selective
Gq-‐inhibitor
FR900359
Raphael
Reher1,
Isabella
Schamari1,
Stefan
Kehraus1,
Suvi
Annala1,
Markus
Kuschak2,
Till
Schäberle1,
Max
Crüsemann1,
Aurelien
Carlier3,4,
Leo
Eberl3,
Christa
E.
Müller2,
Evi
Kostenis1,
Gabriele
M.
König1
1Institute
for
Pharmaceutical
Biology,
University
of
Bonn,
D-‐53115
Bonn,
Germany,
2Institute
for
Phar-‐
maceutical
Chemistry
I,
University
of
Bonn,
D-‐53121
Bonn,
Germany,
3Department
of
Plant
and
Microbial
Biology,
University
of
Zurich,
CH-‐8008
Zurich,
Switzerland,
4Department
of
Microbiology,
University
of
Ghent,
BE-‐9000
Gent,
Belgium
The
cyclic
depsipeptide
FR900359
(FR,
1),
a
selective
inhibitor
of
Gαq
proteins
(Ki
=
6.90
±
1.30nM),
is
applied
as
an
extremely
useful
pharmacological
tool.
Additionally,
this
highly
active
molecule
is
evaluated
for
the
treatment
of
pulmonary
diseases
and
melanoma
[1].
FR
is
present
in
the
higher
plant
Ardisia
crenata
[2],
which
is
known
to
harbor
symbiotic
bac-‐
teria
of
the
genus
Burkholderia
[3,5].
In
the
current
study,
the
novel,
closely
related
cyclic
depsipeptide
AC-‐1
(2)
was
isolated
as
a
minor
metabolite
from
A.
crenata.
Structure
elucida-‐
tion
was
performed
by
extensive
2D-‐NMR
and
MS2
analysis.
Despite
only
little
structural
dif-‐
ferences
in
AC-‐1
as
compared
to
FR
(see
figure
below),
its
bioactivity
towards
Gαq
proteins
is
altered.
These
findings
together
with
results
from
the
semi-‐synthetically
derived
FR-‐
Hexanoate
(3)
allowed
us
to
deduce
first
structure-‐activity-‐relationships.
These
cyclic
depsipeptides
show
several
structural
peculiarities,
(i)
rare,
non-‐proteinogenic
amino
acids
with
manifold
N-‐
and
O-‐methylations,
(ii)
multiple
ester
bonds,
and
(iii)
cis-‐
configurated
amide
bonds
[4],
which
makes
their
non-‐ribosomal
and
bacterial
origin
likely.
Indeed,
sequencing
the
genome
of
Candidatus
Burkholderia
crenata,
the
leaf
nodule
symbiont
of
A.
crenata,
revealed
a
non-‐ribosomal
peptide
synthetase
(NRPS)
gene
cluster,
putatively
encoding
FR
biosynthesis.5
First
results
verified
the
deduced
biosynthetic
hypothesis
and
serve
as
basis
for
further
genetic
and
biochemical
studies.
Acknowledgements:
We
thank
Dr.
Marc
Sylvester
for
measuring
Tandem-‐MS
data
and
Nina
Heycke
for
technical
assistance.
DFG
is
acknowledged
for
funding
research
unit
FOR
2372.
Keywords: Endophyte, Gq-‐Inhibitor, FR900359, Cyclic Depsipeptide, NRPS, SAR
References:
[1] Schrage
R,
Schmitz
AL,
Gaffal
E,
Annala
S,
Kehraus
S,
Wenzel
D,
Büllesbach
KM,
Bald
T,
In-‐
oue
A,
Shinjo
Y,
Galandrin
S,
Shridhar
N,
Hesse
M,
Grundmann
M,
Merten
N,
Charpentier
TH,
Martz
M,
Butcher
AJ,
Slodczyk
T,
Armando
S,
Effern
M,
Namkung
Y,
Jenkins
L,
Horn
V,
Stößel
A,
Dargatz
H,
Tietze
D,
Imhof
D,
Galés
C,
Drewke
C,
Müller
CE,
Hölzel
M,
Milligan
G,
Tobin
AB,
Gomeza
J,
Dohlman
HG,
Sondek
J,
Harden
TK,
Bouvier
M,
Laporte
SA,
Aoki
J,
Fleischmann
BK,
Mohr
K,
König
GM,
Tüting
T,
Kostenis
E.
The
experimental
power
of
FR900359
to
study
Gq-‐regulated
biological
processes.
Nat
Commun
2015;
6:
10156.
[2] Fujioka
M,
Koda
S,
Morimoto
Y,
Biemann
K.
Structure
of
FR900359,
a
cyclic
depsipeptide
from
Ardisia
crenata
Sims.
J
Org
Chem
1988;
53:
2820-‐2825.
[3] Ku
C,
Hu
JM.
Phylogenetic
and
cophylogenetic
analyses
of
the
leaf-‐nodule
symbiosis
in
Ardisia
subgenus
Crispardisia
(Myrsinaceae):
Evidence
from
nuclear
and
chloroplast
markers
and
bacterial
rrn
operons.
Int
J
Plant
Sci
2014;
175:
92-‐109.
[4] Miyamae
A,
Fujioka
M,
Koda
S,
Morimoto
Y.
Studies
of
FR900359,
a
novel
cyclic
depsipep-‐
tide
from
Ardisia
crenata
Sims
1989;
5:
873-‐878.
[5] Carlier
A,
Fehr
L,
Pinto-‐Carbó
M,
Schäberle
T,
Reher
R,
Dessein
S,
König
GM,
Eberl
L.
The
genome
analysis
of
Candidatus
Burkholderia
crenata
reveals
that
secondary
metabolism
may
be
a
key
function
of
the
Ardisia
crenata
leaf
nodule
symbiosis.
Environ
Microbiol
2015;
doi:
10.1111/1462-‐2920.13184
P684
Isolation
of
bioactive
secondary
metabolites
from
mangrove
fun-‐
gal
endophytes
using
epigenetic
regulation
Santana
A.
L.
Thomas1,
Renee
Fleming2,
Lindsey
N.
Shaw2,
Bill
J.
Baker1
1Department
of
Chemistry,
2
Cell
Biology,
Microbiology
and
Molecular
Biology
and
Center
of
Drug
Discov-‐
ery and Innovation, 4202 E. Fowler Ave., University of South Florida, Tampa, FL 33620
Fungal
endophytes
have
gained
increasing
interest
over
the
past
two
decades
as
a
source
of
new
bioactive
natural
products.
As
it
meets
a
developing
source
of
secondary
metabolites,
this
makes
it
an
ideal
focal
point
for
new
natural
products.
Small
molecules
such
as
secondary
me-‐
tabolites
are
targets
for
potential
new
drug
candidates,
especially
for
antibiotic
resistant
bac-‐
teria.
Enterococcus
faecium,
Staphylococcus
aureus,
Klebsiella
species,
Acenitobacter
baumanii,
Pseudomonas
aeruginosa,
Enterobacter
species
(ESKAPE)
pathogens
are
classed
as
the
most
infectious
diseases
because
of
the
lack
of
effective
antibiotics
for
these
pathogens.
The
need
for
new
drugs
can
benefit
from
new
methods
such
as
epigenetic
regulation
of
fungal
metabo-‐
lism.
In
this
project,
a
sample
of
a
black
mangrove
leaf
stem
was
plated
on
Sabouraud
dex-‐
trose
agar
for
isolation
of
the
fungal
endophytes.
Once
isolated
it
was
treated
with
sodium
butyrate,
a
histone
deacetylase
(HDAC)
inhibitor,
resulting
in
a
bioactive
extract
against
methicillin-‐resistant
Staphylococcus
aureus
at
200
µg/mL.
Using
bioactivity-‐guided
fractiona-‐
tion
the
extract
went
through
several
stages
of
separation
beginning
with
normal
phase
me-‐
dium-‐pressure
liquid
chromatography
(MPLC),
then
using
a
flash
column
to
further
purify
the
active
fraction,
which
eluted
on
a
gradient
of
2%
to
12%
ethyl
acetate
in
hexane.
After
obtain-‐
ing
18
fractions
from
the
flash
column,
two
of
the
active
fractions
were
purified
using
high
pressure
liquid
chromatography
(HPLC).
First,
a
normal
phase
preparative
column
was
used
followed
by
a
normal
phase
analytical
column.
The
bioactivity
of
200
µg/mL
was
retained
through
separation.
Keywords: Epigenetic regulation, mangrove endophytic fungi
References:
[1] Aly
A,
Abdessamad
D,
Julia
K,
Peter
P.
Fungal
endophytes
from
higher
plants:
a
prolific
source
of
phytochemicals
and
other
bioactive
natural
products.
Fungal
Diversity
Review
2010;
41:1-‐16
[2] Infectious
Diseases
Society
of
America.
No
ESKAPE!
New
drugs
against
MRSA,
other
su-‐
perbugs
still
lacking.
Science
Daily,
9
December,
2008.
P685
Secondary
metabolites
from
the
fungus
Porodaedalea
pini
Shuen-‐Shin
Yang1,
Hing-‐Yuen
Chan2,
Sung-‐Yuan
Hsieh2,
Gwo-‐Fang
Yuan2,
Su-‐Ling
Wong1,
Chu-‐
Hung
Lin3,
Ming-‐Jen
Cheng2,
Ih-‐Sheng
Chen1,3,4,
Hsun-‐Shuo
Chang1,3,4,5
1Graduate
Institute
of
Natural
Products,
College
of
Pharmacy,
Kaohsiung
Medical
University,
Kaohsuing
807,
Taiwan,
2Bioresource
Collection
and
Research
Center
(BCRC),
Food
Industry
Research
and
Develop-‐
ment
Institute
(FIRDI),
Hsinchu
300,
Taiwan,
3School
of
Pharmacy,
College
of
Pharmacy,
Kaohsiung
Med-‐
ical
University,
Kaohsuing
807,
Taiwan,
4Research
Center
for
Natural
Products
and
Drug
Development,
Kaohsiung
Medical
University,
Kaohsuing
807,
Taiwan.
5Center
for
Infectious
Disease
and
Cancer
Re-‐
search
(CICAR),
Kaohsiung
Medical
University,
Kaohsiung
807,
Taiwan
A
fungus,
Porodaedalea
pini
(Brot.)
Murrill
(Phellinus
pini)
(Hymenochaetaceae),
which
caus-‐
es
tree
diseases
commonly
known
as
“red
ring
rot”
or
“white
speck”,
is
an
important
forest
disturbance
agent
and
plays
a
key
role
in
habitat
formation
for
several
forest
animals.
Previ-‐
ous
research
revealed
the
diverse
constituents
of
P.
pini,
including
amines,
amino
acids,
aro-‐
matic
compounds,
ceramides,
diterpenoids,
lignans,
polyketides,
polysaccharides,
quinones,
and
steroids.
P.
pini
was
processed
through
liquid-‐state
fermentation,
and
its
liquid
fermen-‐
tate
showed
androgen
regulation
(AR)
activity
based
on
a
preliminary
screening.
The
liquid
fermentate
was
partitioned
and
afforded
ethyl
acetate,
n-‐butanol
and
water-‐
soluble
extracts.
Bioassay-‐guided
fractionation
of
the
active
ethyl
acetate-‐soluble
layer
of
the
liquid
fermentate
of
P.
pini
led
to
the
isolation
of
two
new
aromatic
compounds,
piniphenol
A
(1)
and
piniphenol
B
(2),
one
aromatic
compound,
(R)-‐(+)-‐4-‐hydroxyphenyloxirane
(3)
[1]
that
was
isolated
from
nature
for
the
first
time
along
with
five
known
aromatic
compounds,
4-‐
vinylphenol
(4),
4-‐hydroxybenzaldehyde
(5),
2-‐methoxy-‐2-‐(4'-‐hydroxyphenyl)ethanol
(6),
2,3-‐dihydroxypropyl
acetate
(7),
and
phenol
(8).
The
structures
of
these
compounds
were
elucidated
by
spectral
analysis.
The
isolation
of
the
active
fractions
is
still
in
progress
and
the
isolates
are
further
evaluated
with
regard
to
androgen
regulation
activity.
Acknowledgements:
This
work
was
supported
by
Food
Industry
Research
and
Development
Institute
(FIRDI),
Kaohsiung
Medical
University
Research
Foundation
(KMU-‐Q104011).
Keywords: Porodaedalea pini, Hymenochaetaceae, fungus, androgen regulation activity
References:
[1] Sarma
K,
Goswami
A,
Goswami
B.
Exploration
of
chiral
induction
on
epoxides
in
lipase-‐
catalyzed
epoxidation
of
alkenes
using
(2R,3S,4R,5S)-‐(–)-‐2,3:4,6-‐di-‐O-‐isopropylidiene-‐2-‐
keto-‐L-‐gulonic
acid
monohydrate.
Tetrahedron:
Asymmetry
2009;
20:
1295–1300.
P686
Development
of
chemopreventive
agents
for
hepatocellular
car-‐
cinoma
from
Excoecaria
formosana
by
a
glycine
N-‐
methyltransferase
(GNMT)
gene
expression-‐oriented
screen
plat-‐
form
Ho-‐Cheng
Wu1,
Chia-‐Hung
Yen1,2,
Yi-‐Ming
Chen2,
Ya-‐Han
Chang3,
Hsun-‐Shuo
Chang1,2,3,4
1Graduate
Institute
of
Natural
Products,
College
of
Pharmacy,
Kaohsiung
Medical
University,
Kaohsiung
807,
Taiwan,
2
Center
for
Infectious
Disease
and
Cancer
Research
(CICAR),
Kaohsiung
Medical
University,
Kaohsiung
807,
Taiwan,
3School
of
Pharmacy,
College
of
Pharmacy,
Kaohsiung
Medical
University,
Kaohsiung
807,
Taiwan,
4Research
Center
for
Natural
Products
and
Drug
Development,
Kaohsiung
Medi-‐
cal
University,
Kaohsiung
807,
Taiwan
Glycine
N-‐methyltransferase
(GNMT)
is
a
tumor
suppressor
gene
for
hepatocellular
carcino-‐
ma
(HCC)
[1].
The
genotypic
analysis
of
GNMT
may
serve
as
a
marker
for
preventive
and/or
occupational
healthy
consultation.
Approximately
1,400
species
of
Formosan
indigenous
plants
have
been
screened
for
the
GNMT
promotor
activity,
and
the
Excoecaria
formosana
(Hay.)
Hay.
(Euphorbiaceae)
was
found
to
be
one
of
the
most
bioactive
species.
E.
formosana
is
a
shrub
and
mainly
distributed
in
Tonkin,
Indo-‐China,
southern
part
of
Taiwan
in
thickets
and
forests
along
the
seashores
[2].
The
studies
on
Excoecaria
genus
identified
various
classes
of
chemical
constituents,
most
of
which
were
diterpenoids,
flavonoids,
and
galloyl
glucoses.
However,
no
chemical
constituents
and
bioactivity
investigation
of
E.
formosana
have
been
reported.
The
methanolic
extract
of
the
whole
plant
of
E.
formosana
was
partitioned
into
ethyl
acetate,
water,
and
n-‐butanol-‐soluble
layers.
Bioassay-‐guided
fractionation
of
the
active
ethyl
acetate-‐soluble
layer
led
to
the
isolation
of
one
new
steroid:
6'-‐(stigmast-‐5-‐en-‐7-‐hydroperoxy-‐
3-‐O-‐β-‐glucopyransidyl)hexadecanoate
(1)
and
six
known
compounds,
including
four
steroids:
6'-‐(stigmast-‐5-‐en-‐7-‐one-‐3-‐O-‐β-‐glucopyransidyl)hexadecanoate
(2),
(6'-‐O-‐palmitoyl)sitosterol
-‐3-‐O-‐β-‐D-‐glucoside
(3),
a
mixture
of
β-‐sitosterol
(4)
and
stigmasterol
(5),
one
diterpenoid:
ent-‐11α-‐hydroxy-‐3-‐oxo-‐13-‐epi-‐manoyl
oxide
(6),
and
one
benzenoid:
methyl
gallate
(7).
The
successive
isolation
of
the
active
subfractions
of
this
plant
is
still
in
progress
and
the
isolates
are
further
evaluated
with
their
GNMT
promoter
activity
assay.
Acknowledgements:
This
study
was
supported
partially
by
Kaohsiung
Medical
University
“Aim
for
the
Top
Universities
Grant,
grant
No.
KMU-‐TP103H01,
KMU-‐TP103H05
and
KMU-‐TP104E43”
Keywords: Excoecaria formosana, Euphorbiaceae, whole plant, GNMT promoter activity
References:
[1] Chen
YM,
Shiu
JY,
Tzeng
SJ,
Shih
LS,
Chen
YJ,
Lui
WY,
Chen
PH.
Characterization
of
glycine-‐
N-‐methyltransferase-‐gene
expression
in
human
hepatocellular
carcinoma.
Int
J
Cancer
1998;
75:
787−793
[2] Hsieh
CF,
Chaw
SM,
Wang
JC.
Euphorbiaceae.
In:
Flora
of
Taiwan,
Vol.
3,
2nd
edition.
Tai-‐
pei,
Taiwan:
Editorial
Committee
of
the
Flora
of
Taiwan;
1993:
469–470.
P687
Mangrove
associated
fungi
as
a
source
of
potential
drugs
against
the
ESKAPE
pathogens
Sofia
Kokkaliari1,
Renee
Fleeman2,
Lindsey
N.
Shaw2,
Bill
J.
Baker1
Departments
of
1
Chemistry,
2
Cell
Biology,
Microbiology,
and
Molecular
Biology
and
Center
for
Drug
Discovery
and
Innovation,
University
of
South
Florida,
Tampa,
FL
33620,
USA
Over
the
last
decades,
marine
natural
products
have
attracted
the
interest
of
the
scientific
community
due
to
their
chemical
diversity
and
wide
range
of
biological
activities.
Particularly,
marine
fungi
have
been
proven
to
be
a
great
source
of
bioactive
secondary
metabolites.
Man-‐
grove
forests
lay
at
the
intersection
of
land
and
sea,
hosting
both
terrestrial
and
marine
fungal
species.
Due
to
the
competitive
environment
of
this
ecosystem,
fungi
produce
chemical
com-‐
pounds
which
have
been
shown
to
exhibit
biological
activities
[1].
The
ESKAPE
pathogens
(Enterococcus
faecium,
Staphylococcus
aureus,
Klebsiella
pneumoniae,
Acinetobacter
bau-‐
mannii,
Pseudomonas
aeruginosa,
and
Enterobacter
species)
have
been
the
cause
of
many
hos-‐
pital-‐acquired
infections.
Therefore,
the
aim
of
this
project
is
to
isolate
compounds
with
po-‐
tential
activity
against
drug
resisting
bacteria,
such
as
the
ESKAPE
pathogens.
In
particular,
the
fungi
collected
from
the
mangrove
forests
were
grown
under
control
and
epigenetically
modified
conditions,
in
order
to
counterbalance
the
non-‐competitive
laboratory
conditions
[2].
A
fungal
sample,
identified
as
Fusarium
sp.,
was
cultured
on
rice
and
then
extracted
with
methanol
and
ethyl
acetateand
ethyl
acetate,
then
partitioned
between
water
and
hexane,
then
ethyl
acetate.
The
crude
ethyl
acetate
fraction
was
subjected
to
MPLC
(Medium
Pressure
Liquid
Chromatography).
Bioassay–guided
isolation
through
HPLC
(High
Performance
Liquid
Chromatography)
resulted
in
the
isolation
of
three
pure
compounds
with
moderate
to
good
activity,
ranging
between
1
μg/ml
to
50
μg/ml,
against
MRSA
(Methicillin-‐resistant
Staphylo-‐
coccus
aureus).
The
structures
of
the
pure
compounds,
which
belong
to
the
terpene
family,
were
elucidated
by
1D
and
2D
NMR
spectroscopy.
Acknowledgements:
We
are
thankful
for
the
financial
support
received
from
NIH
(AI103715)
as
well
as
from
the
State
of
Florida
for
the
Center
of
Excellence
funding
of
Center
for
Drug
Discovery
and
Innova-‐
tion.
Danielle
Demers
and
Matt
Knestrick,
with
the
assistance
of
innumerable
undergraduate
researchers,
are
thanked
for
early
screening
efforts
that
identified
the
active
fungus.
Keywords:
Epigenetic
modification,
drug
discovery,
secondary
metabolites
References:
[1] Xu
L,
Meng
W,
Cao
C,
Wang
J,
Shan
W,
Wang
Q.
Antibacterial
and
antifungal
compounds
from
marine
fungi.
Mar
Drugs
2015;
13:
3479-‐3513
[2] Beau
J,
Mahid
N,
Burda
W.N,
Harrington
L,
Shaw
LN,
Mutka
T,
Kyle
D.E,
Barisic
B,
van
Ol-‐
phen
A,
Baker
B.J.
Epigenetic
tailoring
for
the
production
of
anti-‐infective
cytosporones
from
the
marine
fungus
Leucostoma
persoonii.
Mar
Drugs
2012;
10:
762-‐774
P688
New
bioactive
secondary
metabolites
from
dung
beetle-‐
associated
bacteria
Soohyun
Um1,
Ki-‐Bong
Oh2,
Jongheon
Shin1,
Dong-‐Chan
Oh1
1
College
of
Pharmacy,
Seoul
National
University,
Gwanak-‐ro
1,
Gwanak-‐gu,
Seoul
08826,
Republic
of
Ko-‐
rea;
2
Department
of
Agricultural
Biotechnology,
College
of
Agriculture
&
Life
Science,
Seoul
National
University,
1
Gwanak-‐ro,
Gwanak-‐gu,
Seoul
08826,
Republic
of
Korea
Insect-‐microbial
symbioses
are
increasingly
recognized
as
ubiquitous
phenomena
in
insect
ecology,
possibly
driving
co-‐evolution
[1].
Secondary
metabolites
produced
by
insect-‐
associated
bacteria
could
play
important
roles
by
mediating
symbioses.
Structural
novelty
and
biological
activity
of
symbiotic
bacterial
metabolites
notably
encouraged
to
study
these
tremendous
chemical
sources
for
biomedical
purpose
[2].
In
our
investigation,
we
isolated
an
identical
bacterial
strain,
belonging
to
the
genus
Streptomyces,
from
the
dung
beetle
Copris
tripartitus
inhabiting
Jeju
Island
three
times
between
2012
and
2014.
This
bacterial
strain
SNU607
produced
two
oxindole
naphtalenediones
which
were
previously
unreported.
The
planar
structures
of
the
compounds
were
determined
by
UV,
MS
spectral
data
and
1D/2D
NMR
spectroscopic
techniques,
while
the
absolute
configurations
of
the
compounds
were
de-‐
termined
by
ECD.
These
two
novel
compounds,
coprisidins
A
and
B
(1
and
2),
were
presuma-‐
bly
produced
through
an
unusual
biosynthesis
pathway.
Coprisidins
A
and
B
inhibit
the
action
of
Na+/K+
ATPase
(IC50
of
1
=
8.95
µg/mL)
and
of
Staphylococcus
aureus
sortase
A
(IC50
of
2
=
32.9
µg/mL).
We
also
isolated
three
more
putatively
associated
bacterial
strains
from
the
dung
beetle’s
gut
(SNU471,
SNU502,
and
SNU533
[3])
producing
new
peptide
class
com-‐
pounds.
Here
we
report
the
dung
beetle-‐associated
bacterial
isolation,
the
structural
elucida-‐
tion
of
their
secondary
metabolites,
and
the
biological
activities
of
the
compounds.
HO HO
O O
NH HO NH
HO HO
O O
OH OH
O O O O
coprisidin A
coprisidin B
Acknowledgements:
This
work
was
supported
by
a
National
Research
Foundation
of
Korea
(NRF)
grant
funded
by
the
Korean
government
(Ministry
of
ICT
and
Future
Planning)
(2014R1A2A1A11053477
and
2009-‐0083533)
Keywords: Symbiotic bacteria, dung beetle, naphtalenedione, peptide
References:
[1] Douglas
AE.
Lessons
from
studying
insect
symbioses.
Cell
Host
Microbe
2011;
10:
359-‐
367
[2] Cragg
GM,
Newman
DJ.
Natural
products:
A
continuing
source
of
novel
drug
leads.
Bio-‐
chim
Biophys
Acta
2013;
1830:
3670-‐3695
[3] Um
S,
Park
SH,
Kim
J,
Park
HJ,
Ko
K,
Bang
HS,
Lee
SK,
Shin
J,
Oh
D-‐C.
Coprisamides
A
and
B,
new
cyclic
peptides
from
a
gut
bacterium
of
the
dung
beetle
Copris
tripartitus.
Org
Lett
2016;
17:
1272-‐1275
P689
Novel
insights
into
plant-‐endophyte
communication:
maytansine
as
an
example
Souvik
Kusari1,
Parijat
Kusari2,
Dennis
Eckelmann1,
Sebastian
Zühlke1,
Oliver
Kayser2,
Michael
Spiteller1
1Institute
of
Environmental
Research
(INFU),
Department
of
Chemistry
and
Chemical
Biology,
Chair
of
Environmental
Chemistry
and
Analytical
Chemistry,
TU
Dortmund,
Otto-‐Hahn-‐Str.
6,
44221
Dortmund,
Germany,
2Department
of
Biochemical
and
Chemical
Engineering,
Chair
of
Technical
Biochemistry,
TU
Dortmund,
Emil-‐Figge-‐Str.
66,
44227
Dortmund,
Germany
Studies
on
microbe-‐host
interactions
in
plant
and
animal
systems
aimed
at
understanding
the
role
of
these
associations
and
their
utility
in
pharmaceutical
and
agricultural
sectors
are
gain-‐
ing
impetus
[1].
Several
recent
studies
have
lent
evidence
to
the
fact
that
certain
so-‐called
“plant
metabolites”
are
actually
biosynthesized
by
associated
endophytic
microorganisms
[2,3].
Given
the
central
role
of
chemical
crosstalk
in
plants
and
associated
endophytes
[see
Figure
(A)],
it
is
essential
to
unravel
the
factors
that
affect
the
nature,
distribution
and
amount
of
“communication”
molecules
at
the
plant-‐microbe
and
microbe-‐microbe
interface
to
gain
fundamental
insights
on
endophytic
biosynthetic
pathways
in
distinct
ecological
niches.
We
recently
investigated
biosynthesis
of
the
important
anticancer
and
cytotoxic
compound
maytansine
in
Celastraceae
plants
in
order
to
elucidate
its
actual
producer(s),
which
has
been
an
open
question
since
its
discovery
in
the
1970s.
We
showed
that
maytansine
is
actually
a
biosynthetic
product
of
root-‐associated
endophytic
bacterial
community
in
Putterlickia
verru-‐
cosa
and
Putterlickia
retrospinosa
plants
[4].
This
extremely
interesting
outcome
provided
the
scientific
basis
to
investigate
the
actual
producer(s)
responsible
for
maytansine
biosynthesis
in
Maytenus
plants.
Endophytic
communities
harboring
different
tissues
of
Maytenus
serrata
originating
from
Cameroon
were
investigated
using
a
combination
of
bioanalytical
tools
such
as
HPLC-‐HRMSn
and
MALDI-‐MSI,
and
targeted
genome
mining
techniques
to
elucidate
the
source
and
sites
of
maytansine
biosynthesis.
We
proved
that
the
biosynthesis
of
maytansine
in
M.
serrata
is
shared
between
the
endophytic
bacterial
community
colonizing
the
stem
and
the
host
plant
containing
non-‐culturable
cryptic
endophytes
[see
Figure
(B)]
[5].
Our
work
demonstrates
that
maytansine
is
biosynthesized
in
M.
serrata
only
when
the
host
plant
joins
forces
with
its
selected
and
very
eco-‐specific
endophytic
bacterial
community.
Acknowledgements:
This
work
was
funded
by
the
“Welcome
to
Africa”
initiative
of
the
German
Federal
Ministry
of
Education
and
Research
(BMBF)
and
German
Academic
Exchange
Service
(DAAD).
The
Minis-‐
try
of
Innovation,
Science,
Research
and
Technology
of
the
State
of
North
Rhine-‐Westphalia,
Germany,
and
the
German
Research
Foundation
(DFG)
are
thankfully
acknowledged
for
granting
a
high-‐resolution
mass
spectrometer.
Keywords: Endophytes, endophytic bacterial community, cross-‐species biosynthesis, may-‐
tansine, plant-‐endophyte communication
References:
[1] Kusari
S,
Hertweck
C,
Spiteller
M.
Chemical
ecology
of
endophytic
fungi:
origins
of
second-‐
ary
metabolites.
Chem
Biol
2012;
19:
792-‐798.
[2] Kusari
S,
Spiteller
M.
Are
we
ready
for
industrial
production
of
bioactive
plant
secondary
metabolites
utilizing
endophytes?
Nat
Prod
Rep
2011;
28:
1203-‐1207.
[3] Kusari
S,
Spiteller
M.
Metabolomics
of
endophytic
fungi
producing
associated
plant
sec-‐
ondary
metabolites:
progress,
challenges
and
opportunities.
In:
Roessner
U,
editor.
Metabolomics.
Rijeka,
Croatia:
InTech,
2012;
241-‐266.
[4] Kusari
S,
Lamshöft
M,
Kusari
P,
Gottfried
S,
Zühlke
S,
Louven
K,
Hentschel
U,
Kayser
O,
Spiteller
M.
Endophytes
are
hidden
producers
of
maytansine
in
Putterlickia
roots.
J
Nat
Prod
2014;
77:
2577-‐2584.
[5] Kusari
P,
Kusari
S,
Eckelmann
D,
Zühlke
S,
Kayser
O,
Spiteller
M.
Cross-‐species
biosynthe-‐
sis
of
maytansine
in
Maytenus
serrata.
RSC
Adv
2016;
6:
10011-‐10016.
P690
A
comparison
between
the
application
of
NMR
and
LC-‐HRMS
based
metabolomics
on
the
discovery
of
natural
products
from
endophytic
fungi
Trevor
N.
Clark1,
Fabrice
Berrué2,
Larry
Calhoun3,
Patricia
Boland4,5,
Russel
Kerr4,5,
John
A.
Johnson1,
Christopher
A.
Gray
1,3
1Department
of
Biological
Sciences,
University
of
New
Brunswick,
100
Tucker
Park
Road,
E2L
4L5
Saint
John,
Canada,
2National
Research
Council
Canada,
1411
Oxford
Street,
B3H
3Z1
Halifax,
Canada,
3Department
of
Chemistry,
University
of
New
Brunswick,
30
Dineen
Drive,
E3B
5A3
Fredericton,
Canada,
4Department
of
Chemistry,
Atlantic
Veterinary
College,
University
of
Prince
Edward
Island,
550
University
Avenue,
C1A
4P3
Charlottetown,
Canada,
5Department
of
Biomedical
Sciences,
Atlantic
Veterinary
Col-‐
lege,
University
of
Prince
Edward
Island,
550
University
Avenue,
C1A
4P3
Charlottetown,
Canada
Natural
products
have
been
an
excellent
source
of
diverse
and
complex
chemical
structures.
However,
as
we
isolate
more
natural
products
it
becomes
increasingly
difficult
to
discover
novel
chemical
entities.
Screening
strategies
that
rely
solely
on
evaluation
of
biological
activi-‐
ty
are
becoming
redundant,
and
metabolomic
approaches
are
being
increasingly
used
to
avoid
the
isolation
of
known
natural
products
[1].
In
this
project,
we
employed
nuclear
magnetic
resonance
(NMR)
[2]
as
well
as
liquid
chromatography
tandem
high
resolution
mass
spec-‐
trometry
(LC-‐HRMS)-‐based
[3]
metabolomics
screens
in
an
effort
to
more
effectively
priori-‐
tise
endophyte
extracts
for
natural
product
discovery.
Fungi
used
in
this
project
were
isolated
from
the
leaves
of
medicinal
plants
used
by
the
Canadian
First
Nations.4
Independent
princi-‐
pal
component
analysis
of
the
NMR
and
LC-‐HRMS
metabolomics
data
identified
extracts
pos-‐
sessing
unique
metabolic
profiles
from
the
two
screening
methods
and
these
extracts
were
selected
for
further
investigation.
The
NMR
metabolomic
screening
prioritised
five
extracts
that
contained
a
high
abundance
of
structurally
similar
natural
products,
particularly
those
showing
an
abundance
of
peaks
between
1.0-‐2.0
ppm
and
6.0-‐6.4
ppm,
and
ultimately
led
to
the
isolation
of
only
known
compounds.
Advantages
gained
through
the
increased
sensitivity
and
accuracy
of
LC-‐HRMS
make
the
mass
spectrometric
based
method
greatly
superior
as
a
screening
and
prioritization
tool.
LC-‐HRMS
based
metabolomics
strength
lies
within
the
completeness
of
the
chemical
database
that
is
being
used
as
a
dereplication
step.
Prioritized
extracts
can
be
removed
from
further
study
if
the
target
m/z
chosen
by
PCA
has
a
known
nat-‐
ural
product
structure
within
5ppm.
This
strategy
of
prioritization
has
highlighted
seven
en-‐
dophytic
extracts
containing
an
m/z
without
a
match
in
our
database
allowing
for
the
discov-‐
ery
of
new
natural
products
from
our
fungal
library
in
the
future.
Acknowledgements: Hebelin Correa is acknowledged for technical assistance
Keywords: Endophytic fungi, medicinal plants, metabolomics, NMR, LC-‐HRMS
References:
[1] Wu
C,
Kim
HK,
van
Wezel
GP,
Choi
YH.
Metabolomics
in
the
natural
products
field
–
a
gateway
to
novel
antibiotics.
Drug
Discov
Today
Technol
2015;
13:
11−17
[2] Clark
TN,
Bishop
AI,
McLaughlin
M,
Calhoun
LA,
Johnson
JA,
Gray
CA.
Isolation
of
(-‐)-‐
avenaciolide
as
the
antifungal
and
antimycobacterial
constituent
of
a
Seimatosporium
sp.
endophyte
from
the
medicinal
plant
Hypericum
perforatum.
Nat
Prod
Comm
2014;
9:
1495−1496
[3] Forner
D,
Berrué
F,
Correa
H,
Duncan
K,
Kerr
RG.
Chemical
dereplication
of
marine
acti-‐
nomycetes
by
liquid
chromatography-‐high
resolution
mass
spectrometry
profiling
and
statistical
analysis.
Anal
Chim
Acta
2013;
805:
70−79
[4] Ellsworth
KT,
Clark
TN,
Gray
CA,
Johnson
JA.
Isolation
and
bioassay
screening
of
medicinal
plant
endophytes
from
eastern
Canada.
Can
J
Microbiol
2013;
59:
761−765
P691
Unusual
natural
products
mediated
root
hairs-‐endophyte
stack-‐
ing
(RHESt)
that
traps
and
kills
pathogens
Walaa
K.
Mousa1,
Charles
Shearer2,
Cassandra
Ettinger3,
Jonathan
Eisen3,
Manish
N.
Raizada2
1
Department
of
Biochemistry
and
Biomedical
Sciences,
McMaster
University,
Hamilton,
ON
Canada
L8S
4L8;
2
Department
of
Plant
Agriculture,
University
of
Guelph,
Guelph,
ON
Canada
N1G
2W1,
3
University
of
California
Davis
Genome
Center,
Davis,
California,
USA
95616
The
ancient
African
crop,
finger
millet,
has
broad
resistance
to
pathogens
including
the
toxi-‐
genic
fungus
Fusarium
graminearum
[1].
Here
we
report
the
discovery
of
a
novel
plant
de-‐
fence
mechanism,
resulting
from
an
unusual
symbiosis
between
finger
millet
and
a
root-‐
inhabiting
bacterial
endophyte,
M6
(Enterobacter
sp.)
[2].
Upon
sensing
the
pathogen,
M6
swarms
towards
Fusarium
attempting
to
penetrate
root
epidermis,
induces
growth
of
root
hairs
through
production
of
growth
hormones,
which
then
bend
parallel
to
the
root
axis,
then
forms
biofilm-‐mediated
microcolonies,
resulting
in
a
remarkable,
multi-‐layer
root
hair-‐
endophyte
stack
(RHESt).
The
RHESt
consists
of
two
lines
of
defence,
a
dense
layer
of
interca-‐
lated
root
hairs
and
endophyte
microcolonies
followed
by
a
long,
continuous
endophyte
bar-‐
rier
layer
on
the
root
epidermal
surface
(see
A
and
B).
M6
was
visualized
to
specifically
attach
to
Fusarium
hyphae
(see
C)
and
kill
it
(see
D).
RHESt
results
in
a
physical
barrier
that
prevents
entry
and/or
traps
F.
graminearum
which
is
then
killed
inside.
Thus
M6
creates
its
own
spe-‐
cialized
killing
microhabitat.
M6
killing
requires
c-‐di-‐GMP-‐dependent
signalling,
diverse
fun-‐
gicides
(phenazine
metabolites,
colicin
V
peptide
antibiotic,
chitinase
enzyme,
etc.)
[3,
4].
We
show
that
a
pre-‐requisite
of
M6
killing
is
expression
of
a
fusaric
acid
resistance
operon
that
effluxes
the
Fusarium-‐derived
mycotoxin,
fusaric
acid,
outside
the
cell
(see
E).
Furthermore,
our
results
show
a
novel
epistatic
regulatory
interaction
between
the
fusaric
acid
resistance
and
phenazine
pathways.We
propose
that
the
phenazine-‐fusaric
acid
arms
race
provides
a
molecular
and
biochemical
paleontological
record
that
M6
and
Fusarium
co-‐evolved.
The
end-‐
result
of
this
remarkable
symbiosis
and
tripartite
co-‐evolution
is
reduced
DON
mycotoxin
[5],
potentially
benefiting
millions
of
subsistence
farmers
over
thousands
of
years.
RHESt
demon-‐
strates
the
value
of
exploring
ancient,
orphan
crop
endophytic
microbiomes.
E
Acknowledgements:
We
thank
Dr.
Michaela
Strueder-‐Kypke
for
technical
assistance.
We
thank
the
graphical
designer,
Lisa
Smith
for
the
graphical
illustration.
Keywords: Endophytes, finger millet, F. graminearum, RHESt, antifungal natural products
References:
[1] Mousa
W,
Schwan
A,
Davidson
J,
Strange
P,
Liu
H,
Zhou
T,
Auzanneau
FI,
Raizada
M.
An
en-‐
dophytic
fungus
isolated
from
finger
millet
(Eleusine
coracana)
produces
anti-‐fungal
nat-‐
ural
products.
Front
Microbiol
2015;
6:
1157−1173
[2] Ettinger
C,
Mousa
W,
Raizada
M,
Eisen
J.
Draft
genome
sequence
of
Enterobacter
sp.
str.
UCD-‐UG_FMILLET
(Phylum
Proteobacteria).
Genome
Announce
2015;
3:
1461−1462
[3] Mousa
W,
Raizada
M.
The
diversity
of
anti-‐microbial
secondary
metabolites
produced
by
fungal
endophytes:
An
interdisciplinary
perspective.
Front
Microbiol
2016;
4:
65−83
[4] Mousa
W,
Raizada
N.
Biodiversity
of
genes
encoding
anti-‐microbial
traits
within
plant
as-‐
sociated
microbes.
Front
Plant
Sci
2015;
6:
231−256
[5] Mousa
W,
Shearer
C,
Limay-‐Rios
V,
Zhou
T,
Raizada
M.
Bacterial
endophytes
from
wild
maize
suppress
Fusarium
graminearum
in
modern
maize
and
inhibit
mycotoxin
accumu-‐
lation.
Front
Plant
Sci
2015;
6:
805−824
P693
Search
for
anti-‐dengue
secondary
metabolites
from
two
Diospy-‐
ros
endophytes,
Aspergillus
sp.
and
Phomopsis
sp.
Laure-‐Anne
Peyrat1,
Véronique
Eparvier1,
Cécilia
Eydoux2,
Jean-‐Claude
Guillemot2,
Didier
Stien3,
Marc
Litaudon1
1
Institut
de
Chimie
des
Substances
Naturelles,
CNRS,
UPR2301,
University
Paris-‐Saclay,
91198-‐Gif-‐sur-‐
Yvette
Cedex,
France,
2
Centre
de
Recherche
Architecture
et
Fonction
des
Macromolécules
Biologiques,
UMR
7257
CNRS
-‐
Aix-‐Marseille
Univ.,
163
Avenue
de
Luminy,
13288-‐Marseille
Cedex
09,
France,
3
Sor-‐
bonne
Universités,
UPMC
Univ
Paris
06,
CNRS,
Laboratoire
de
Biodiversité
et
Biotechnologies
Micro-‐
biennes
(LBBM),
Observatoire
Océanologique,
66650-‐Banyuls-‐sur-‐mer,
France
Dengue
fever
is
the
most
prevalent
mosquito-‐borne
viral
disease
of
humans
with
an
estimated
390
million
infections
occurring
annually
[1].
A
new
dengue
vaccine
has
been
recently
regis-‐
tered
in
several
countries,
but
no
specific
antiviral
therapy
is
available
for
the
treatment
of
this
disease.
A
screening,
using
a
dengue
replicon
virus-‐cell-‐based
assay
[2],
was
performed
on
3563
ethyl
acetate
extracts
from
1500
plants
and
led
to
the
selection
of
25
tropical
species
from
the
genus
Diospyros
(Ebenaceae)
for
their
significant
activity.
The
aim
of
the
study
is
to
isolate
and
characterize
active
compounds
against
dengue
virus
rep-‐
lication
from
Diospyros
species
and
their
associated
endophytes.
The
endophytes
of
2
active
plants,
D.
ierensis
and
D.
carbonaria
(F.
Guiana)
were
isolated
and
identified
[3].
A
total
of
82
strains,
corresponding
to
11
fungal
genera,
were
identified
and
their
extracts
were
tested
against
dengue
virus
replication.
The
most
active
endophytes,
Aspergillus
sp.
(LAP1-‐7-‐57)
and
Phomopsis
sp.
(LAP1-‐7-‐32),
were
investigated.
Three
pyranones,
2
dehydro-‐aflavinine
derivatives
(1
new),
betulinic
acid,
a
new
steroid
named
gilvsin
E,
and
a
series
of
12
new
trans-‐decaline
carneic
acid
analogues
were
isolated.
All
these
molecules
will
be
tested
to
evaluate
their
dengue
virus
inhibition.
It’s
the
first
time
that
betulinic
acid
has
been
isolated
from
an
endophyte.
It
should
be
mentioned
that
this
molecule
has
been
isolated
from
the
host
plant
(D.
carbonaria),
and
biological
assay
showed
a
significant
inhibitory
activity
on
dengue
virus
replication.
Our
work
demonstrates
that
this
endophyte
is
able
to
synthesize
the
same
active
molecule
than
its
host
plant.
A
gene
transfer
from
the
host
plant
to
its
endophyte
might
have
occurred.
Acknowledgements:
This
work
has
benefited
from
an
“Investissement
d’Avenir”
grant
managed
by
ANR
(CEBA,
ref
ANR-‐10-‐LABX-‐0025)
Keywords: Dengue replicon, Diospyros spp., endophytes, Aspergillus sp., Phomopsis sp.
References:
[1] Behnam
MAM,
Nitsche
C,
Boldescu
V,
Klein
CD.
The
Medicinal
Chemistry
of
Dengue
Virus
Med
Chem
2016;
advance
online
publication
19
May
2016;
DOI:
10.1021/acs.jmedchem.5b01653
[2] Massé
N,
Davidson
A,
Ferron
F,
Alvarez
K,
Jacobs
M,
Romette
JL,
Canard
B,
Guillemot
JC.
Dengue
virus
replicons:
production
of
an
interserotypic
chimera
and
cell
lines
from
dif-‐
ferent
species,
and
establishment
of
a
cell-‐based
fluorescent
assay
to
screen
inhibitors,
validated
by
the
evaluation
of
ribavirin's
activity.
Antiviral
Res
2010;
86:
296–305.
[3] Kusari
S,
Hertweck
C,
Spiteller
M.
Chemical
ecology
of
endophytic
fungi:
origins
of
second-‐
ary
metabolites.
Chem
Biol
2012;
19:
792–798.
P694
Anti-‐MRSA
natural
products
from
an
epigenetic
modified
Floridi-‐
an
mangrove-‐associated
fungus
Sylvia
Soldatou1,
2,
Renee
Fleeman3,
Lindsey
N.
Shaw3,
Bill
J.
Baker2
1School
of
Chemistry,
National
University
of
Ireland,
Galway,
Ireland,
Departments
of
2Chemistry
and
3Cell
Biology,
Microbiology,
and
Molecular
Biology
and
the
Center
for
Drug
Discovery
and
Innovation
(CDDI), University of South Florida, Tampa, FL 33620, USA.
The ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae,
Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.) are responsible for
the majority of nosocomial infections and are resistant to most antibacterial agents [1]. The
continuous need for new antibiotics has invigorated drug discovery research, leading us to
investigate niche biological sources for new chemodiversity. We focus on mangrove-‐
associated microorganisms which have been found to be a great source of chemically diverse
and biologically active metabolites [2]. In particular, we have been interested in the isolation
and characterisation of secondary metabolites from Floridian mangrove-‐associated fungi as
well as their biological activity against the ESKAPE pathogens. Moreover, we are interested in
enhancing the bioactivity of these secondary metabolites by epigenetic regulation of the fungi
using histone deacetylase (HDAC) and DNA methyltransferase (DNMT) inhibitors [3].
During the course of this work, a suite of new and known compounds with anti-‐MRSA activity
have been uncovered. These compounds are produced by an endophytic fungus derived from
the leaves of the Floridian red mangrove. Using an enhanced rice medium, the fungus was cul-‐
tured under the regulation of an HDAC inhibitor. After a 21-‐day incubation period the fungal
culture was extracted 3 x with EtOAc and the crude extract was partitioned between water
and EtOAc as shown in Figure 1.
Figure
1:
Isolation
scheme
of
anti-‐MRSA
compounds
Following
the
anti-‐MRSA
activity,
the
EtOAc
partition
was
active
against
MRSA
at
5
µg/ml
and
was
subjected
to
MPLC
giving
five
fractions.
All
bioactive
MPLC
fractions
were
purified
through
several
rounds
of
HPLC
which
led
to
the
isolation
of
at
least
four
new
and
know
com-‐
pounds,
exhibiting
moderate
activity
against
MRSA.
The
structures
of
the
pure
compounds,
which
belong
to
the
benzopyrone
and
benzofuran
families
were
elucidated
by
1D
and
2D
NMR
spectroscopy.
Acknowledgements:
We
acknowledge
Danielle
H.
Demers,
Matthew
A.
Knestrick
and
all
undergraduates
who
worked
on
the
small-‐scale
high
throughput
screening
fungal
project.
We
are
also
grateful
for
finan-‐
cial
support
from
the
NIH
(AI103715),
as
well
as
the
State
of
Florida
for
Center
of
Excellence
funding
of
CDDI.
Keywords: mangrove, fungi, bioassay-‐guided isolation, MRSA, epigenetic regulation
References:
[1] Boucher
WH,
Talbot
HG,
Bradley
SJ,
Edwards
EJ,
Gilbert
D,
Rice
BL,
Scheld
M,
Brad
Spellberg
B,
John
Bartlett
J.
Bad
Bugs,
No
Drugs:
No
ESKAPE!
An
Update
from
the
Infectious
Diseas-‐
es
Society
of
America.
Clin
Infect
Dis
2009;
48:
1-‐12
[2] Xu
J.
Bioactive
natural
products
derived
from
mangrove-‐associated
microbes.
RSC
Ad-‐
vances
2015;
5:
841-‐892
[3] Beau
J,
Mahid
N,
Burda
NW,
Harrington
L,
Shaw
NL,
Mutka
T,
Kyle
ED,
Barisic
B,
van
Olphen
A,
Baker
JB.
Epigenetic
tailoring
for
the
production
of
anti-‐infective
cytosporones
from
the
marine
fungus
Leucostoma
persoonii.
Mar
Drugs
2012;
10:
762-‐774
…
P695
Bioinspired
total
syntheses
of
(±)-‐dictazole
B
and
(±)-‐
tubastrindole
B:
"The
aplysinopsins’
cascade"
Adam
Skiredj1,
Mehdi
A.
Beniddir1,
Delphine
Joseph1,
Guillaume
Bernadat1,
Laurent
Evanno1,
Erwan
Poupon1
1
BioCIS,
Univ.
Paris-‐Sud,
CNRS,
LabEx
LERMIT,
Université
Paris-‐Saclay,
92296
Châtenay-‐Malabry,
France
When
trying
to
figure
out
the
natural
origin
of
the
aplysinopsin
family
of
alkaloids,
one
essen-‐
tial
feature
emerges
from
the
isolation
reports.
All
the
congeners
of
this
series
are
indeed
coming
from
sponges
or
stony
corals
collected
in
shallow
waters,
which
means
within
reach
of
sunlight.
Three
dinstinct
scaffolds
are
represented
in
the
family
and,
on
a
structural
basis,
a
dimerization
step
is
obviously
needed
at
some
point
to
form
the
more
challenging
dictazole-‐
type
and
tubastrindole-‐type
alkaloids.
Guided
by
these
biosynthetic
considerations,
we
have
carried-‐out
the
first
total
synthesis
of
(±)-‐dictazole
B
[1]
through
a
diastereoselective
[2+2]
photocycloaddition
between
two
aplysinopsin-‐type
monomers.
Beyond
this
surprisingly
sim-‐
ple
and
efficient
synthesis,
we
then
considered
dictazole-‐type
compounds
as
possible
inter-‐
mediates
toward
their
tubastrindole-‐type
relatives.
This
fruitful
ring
expansion
approach
led
to
the
first
total
synthesis
of
(±)-‐tubastrindole
B
[2]
with
full
retention
of
configuration
during
the
cascade.
Given
these
results,
one
could
imagine
that
the
chosen
centrosymmetric
dicta-‐
zole-‐type
precursors
might
exist
in
nature
even
if
they
have
not
been
isolated
to
date.
Our
work
has
also
resulted
in
the
synthesis
of
a
transient
intermediate
explaining
the
formation
of
other
natural
products
of
the
series.
Moreover,
the
extended
study
uncovered
a
synthetic
network
around
these
marine
alkaloids
[3].
Keywords:
Total
synthesis,
Biomimetic
synthesis,
Alkaloids,
Self-‐assembly
References:
[1] Skiredj
A,
Beniddir
MA,
Joseph
D,
Leblanc
K,
Bernadat
G,
Evanno
L,
Poupon
E.
Spontaneous
biomimetic
formation
of
(±)-‐dictazole
B
under
irradiation
with
artificial
sunlight.
Angew
Chem
Int
Ed
2014;
53:
6419-‐6424
[2] Skiredj
A,
Beniddir
MA,
Joseph
D,
Leblanc
K,
Bernadat
G,
Evanno
L,
Poupon
E.
A
unified
bio-‐
inspired
“Aplysinopsin
cascade”:
total
synthesis
of
(±)-‐tubastrindole
B
and
related
bio-‐
synthetic
congeners.
Org
Lett
2014;
16:
4980-‐4983
[3] Skiredj
A,
Beniddir
MA,
Joseph
D,
Bernadat
G,
Evanno
L,
Poupon
E.
Harnessing
the
intrinsic
reactivity
within
the
aplysinopsin
series
for
the
synthesis
of
intricate
dimers:
natural
from
start
to
finish.
Synthesis
2015;
47:
2367-‐2376
P696
Harnessing
the
main
event
of
drimentines
biosynthesis:
bio-‐
inspired
synthesis
and
biological
evaluation
of
Δ8’-‐isodrimentine
A
and
related
compounds
Adam Skiredj, Mehdi A. Beniddir, Laurent Evanno, Erwan Poupon
BioCIS, Univ. Paris-‐Sud, CNRS, LabEx LERMIT, Université Paris-‐Saclay, 92296 Châtenay-‐Malabry, France
Drimentines
are
a
family
of
alkaloids
biosynthetically
originating
from
the
condensation
of
a
sesquiterpene
unit
onto
a
tryptophan-‐containing
cyclic
dipeptide
[1].
These
alkaloids
feature
a
fused
diketopiperazino-‐pyrroloindoline
core.
Their
scaffold
is
likely
to
arise
from
an
indol-‐
dearomatization
followed
by
the
intramolecular
trapping
of
an
indolenine
intermediate.
The
aim
of
the
study
was
to
exploit
this
biosynthetic
hypothesis
to
develop
a
straightforward
as-‐
sembly
of
the
drimentine
A
core.
This
concise
synthetic
sequence
is
described
herein.
Our
du-‐
al
strategy
involves
the
alkylation
of
a
tryptophane-‐containing
cyclodipeptide
by
a
drimane-‐
type-‐decaline
or
a
linear
C15
unit.
In
both
approaches,
the
starting
cyclopeptide
was
obtained
by
classical
peptidic
couplings.
The
drimane-‐type-‐decaline
was
prepared
from
sclareolide
while
the
fully
biomimetic
approach
was
also
evaluated
with
the
synthesis
of
a
farnesyl-‐
substituted
pyrrolo-‐indoline
diketopiperazine.
That
farnesylated
derivative
is
corresponding
to
a
biosynthetic
precursor
of
drimentines
A
and
D.
The
envisioned
dearomatization-‐trapping
sequence
has
been
successfully
realized
validating
the
synthetic
rational
and
furnishing
bio-‐
synthetic
clues.
Hence,
very
close
analogs
of
the
natural
substances,
such
as
Δ8’-‐isodrimentine
A,
have
been
obtained
and
biologically
evaluated
[2].
Keywords:
Drimentines,
biomimetic
synthesis,
alkaloids,
dearomatization
References:
[1] Che
Q,
Zhu
T,
Qi
X,
Màndi
A,
Kurtàn
T,
Mo
X,
Li
J,
Gu
Q,
Li
D.
Hybrid
isoprenoids
from
a
reeds
rhizosphere
soil
derived
actinomycete
streptomyces
sp.
CHQ-‐64.
Org
Lett
2012;
14:
3438-‐3441
[2] Skiredj
A,
Beniddir
M
A,
Evanno
L,
Poupon
E.
Mimicking
the
main
event
of
drimentines
bio-‐
synthesis:
synthesis
of
Δ8’-‐isodrimentine
A
and
related
compounds,
biological
evaluation.
Eur
J
Org
Chem
Eur.
doi:
10.1002/ejoc.201600444
P697
Enzymatic
tailoring
of
oleuropein
isolated
from
Olea
europaea
leaves
Efstratios
Nikolaivits1,
Aikaterini
Termentzi2,3,
Alexios-‐Leandros
Skaltsounis2,
Nikolas
Fokia-‐
lakis2,
Evangelos
Topakas1
1Biotechnology
Laboratory,
School
of
Chemical
Engineering,
National
Technical
University
of
Athens,
5
Iroon
Polytechniou
Str,
Zografou
Campus,
15700,
Athens
Greece.
2Laboratory
of
Pharmacognosy
and
Natural
Products
Chemistry,
School
of
Pharmacy,
National
and
Kapodistrian
University
of
Athens,
Pane-‐
pistimiopolis
Zografou,
17755,
Athens,
Greece.
3Department
of
Pesticides
Control
and
Phytopharmacy,
Benaki
Phytopathological
Institute,
St.
Delta
8,
14561,
Kifissia,
Greece
Oleuropein
is
one
of
the
main
phenolic
compounds
of
the
olive
plant,
Olea
europaea.
It
can
be
found
in
high
quantities
in
leaves
and
roots
(up
to
500mg/g
dry
extract)
[1].
It
is
a
glucosylat-‐
ed
ester
of
hydroxytyrosol
(HT)
with
the
secoiridoid
elenolic
acid
(EA).
Oleuropein’s
various
pharmacological
properties
have
been
unveiled,
including
antioxidant,
antimicrobial,
cardio-‐
protective,
hypolipidemic
etc.
HT
is
one
of
the
most
important
degradation
products
of
oleuropein,
during
the
fruit
maturation
process.
It
is
a
strong
antioxidant
and
the
only
olive
molecule
with
an
approved
health
claim
from
EFSA
[2].
Other
hydrolysis
products
of
oleuro-‐
pein
are
a
wide
range
of
oleuropein
aglycones
and
EA,
both
possessing
important
biological
activities,
such
as
antioxidant,
anti-‐inflammatory,
antimicrobial,
antiviral.
There
have
been
several
efforts
towards
HT
production
through
oleuropein
hydrolysis
by
chemical
procedures
[3].
However,
the
enzymatic
approach
is
always
preferred,
as
it
results
in
environmentally
friendly
products
through
selective
hydrolysis.
So
far,
different
commer-‐
cial
crude
enzyme
preparations
have
been
used
for
the
hydrolysis
of
oleuropein
to
give
HT,
however
the
bioconversion
could
not
tailor
the
final
products
due
to
many
enzymatic
activi-‐
ties
present
[4].
Furthermore,
a
hyperthermophilic
β-‐glucosidase,
catalyzed
the
hydrolysis
of
the
glycosidic
bond,
releasing
HT
by
the
chemical
hydrolysis
of
the
ester
bond
due
to
the
tem-‐
perature
and
pH
reaction
conditions
[5].
In
the
present
study
the
enzymatic
tailored
hydrolysis
of
oleuropein,
purified
from
Olea
euro-‐
paea
leaves
is
reported.
Utilization
of
a
commercial
lipase
(Lipolase,
Novozymes)
and
a
non-‐
commercial
GH3
β-‐glucosidase
from
Myceliophthora
thermophile
[6]
resulted
in
the
hydrolysis
of
either
the
glycosidic
bond
between
the
glucose
and
the
EA
moiety
or
of
the
ester
bond
be-‐
tween
the
EA
moiety
and
HT.
The
identification
of
the
all
the
hydrolysis
products
was
per-‐
formed
by
LC-‐HRMS/MS
and
1D
&
2D
NMR
experiments.
-glucosidase lipase
Keywords:
oleuropein,
enzymatic
tailoring,
hydroxytyrosol,
elenolic
acid,
oleuropein
agly-‐
cone
References:
[1] Michel
T,
Khlif
I,
Kanakis
P,
Termentzi
A,
Allouche
N,
Halabalaki
M,
Skaltsounis
AL.
UHPLC-‐
DAD-‐FLD
and
UHPLC-‐HRMS/MS
based
metabolic
profiling
and
characterization
of
differ-‐
ent
Olea
europaea
organs
of
Koroneiki
and
Chetoui
varieties.
Phytochem
Lett
2015;
11:
424−439
[2] EFSA
Scientific
Opinion,
Article
13(1)
of
Regulation
(EC)
No
1924/20061,
EFSA
Journal,
2011;
9:
2033.
[3] Walter
WM
Jr,
Fleming
HP,
Etchells
JL.
Preparation
of
antimicrobial
compounds
by
hy-‐
drolysis
of
oleuropein
from
green
olives.
J
Appl
Microbiol
1973;
26:
773−776
[4] Yuan
JJ,
Wang
CZ,
Ye
JZ,
Tao
R,
Zhang
YS.
Enzymatic
hydrolysis
of
oleuropein
from
Olea
Europea
(olive)
leaf
extract
and
antioxidant
activities.
Molecules
2015;
20:
2903−2921
[5] Briante
R,
La
Cara
F,
Febbraio
F,
Barone
R,
Piccialli
G,
Carolla
R,
Mainolfi
P,
De
Napoli
L,
Patumi
M,
Fontanazza
G,
Nucci
R.
Hydrolysis
of
oleuropein
by
recombinant
β-‐glycosidase
from
hyperthermophilic
archaeon
Sulfolobus
solfataricus
immobilised
on
chitosan
ma-‐
trix.
J
Biotechnol
2000;
77:
275−286
[6] Karnaouri
A,
Topakas
E,
Paschos
T,
Taouki
I,
Christakopoulos
P.
Cloning,
expression
and
characterization
of
an
ethanol
tolerant
GH3
β-‐glucosidase
from
Myceliophthora
ther-‐
mophila.
PeerJ
2013;
1:
e46.
P698
HPLC
quantification
method
for
chrysin
and
tectochrysin
in
Flourensia
extracts
M. Ángeles Ramírez-‐Cisneros, Ramiro Ríos Gómez, María Yolanda Rios
Centro
de
Investigaciones
Químicas,
IICBA,
Universidad
Autónoma
del
Estado
de
Morelos.
Av.
Univer-‐
sidad
1001
Col.
Chamilpa
62209
Cuernavaca,
Morelos,
México
Flourensia
genus
(Asteraceae)
is
formed
by
42
species
growing
exclusively
in
American
conti-‐
nent,
adapted
to
arid
conditions
is
able
to
replace
the
typical
desert
vegetation.
Leaves
of
the-‐
se
subshrubs,
shrubs
and
small
trees
are
characterized
by
the
production
and
secretion
of
resins
[1].
Flourensia
species
exhibit
several
protective
properties
as
herbicide,
insecticide,
antibacterial,
antialgal,
antifungal,
antitermite
[1].
F.
resinosa
dominates
widely
the
vegetation
were
it
grows
[2]
and
this
fact
is
related
with
its
chemical
content
which
has
been
reported
previously
[3].
Flavones
chrysin
(1)
and
tectochysin
(2),
besides
sesquiterpene
ilicic
acid
(3)
are
major
metabolites
of
this
species.
Compounds
1-‐3
were
reported
to
show
some
of
the
mentioned
biological
activities
[4-‐6].
In
the
present
work
an
HPLC
method
to
quantify
fla-‐
vones
1-‐2
(270λ)
and
identify
sesquiterpene
3
(235λ)
in
Flourensia
extracts
was
developed
and
validated.
UV-‐HPLC
equipment
with
diode
array
detector
was
used.
Chromatographic
separation
of
analytes
was
performed
using
a
5µm
C18
column
(4.6
id
x
250
mm)
at
20°C.
Mobile
phase
was
a
mixture
of
A
(aqueous
TFA
0.05%)
and
B
(MeOH).
Multigradient
elution
was
carried
out
as
follows:
0-‐4
min
from
100%
to
20%
A,
1
ml/min
flow,
4-‐12
min
20%
A
at
a
0.5
ml/min
flow,
12-‐14
min
from
20%
to
0%
A,
at
min
16
flow
was
increased
to
1
ml/min,
maintained
until
min
20
and
then
returned
to
original
conditions.
Method
display
good
selec-‐
tivity,
accuracy,
precision,
linearity
and
reproducibility
and
was
used
to
analyze
content
of
1-‐
3
from
four
different
Flourensia
species:
F.
resinosa,
F.
cernua,
F.
glutinosa
and
F.
laurifolia
(Table
1).
Species
(1)
(2)
(3)
1.100 ZYEXTRACTOS DIF. FLOURENSIAS II #9 UV_VIS_4
mAU WVL:270 nm
4: Tectocrisina
1: Crisina
F. cernua -‐ -‐ -‐
F. glutinosa -‐ -‐ -‐
F. laurifolia 1.18 -‐ + 500
F. resinosa 29.95 6.47 +
Results represent the mean of triplicate
3
analysis and are expressed as compound (mg)
2
/ dry vegetal material (g). -‐ non detected. + min
-100
identified 0,0 5,0 10,0 15,0 20,0 25,0
Acknowledgements:
CONACyT
(grant
number
241044)
financed
this
work.
Results
were
taken
in
part
from
Fernando
Rodríguez
Garrido
bachelor
thesis
(UAEM,
2015).
Keywords: Chrysin, Tectochrysin, Flourensia, Asteraceae, HPLC quantification
References:
[1] Rios
MY.
Chemistry
and
Biology
of
the
Genus
Flourensia
(Asteraceae).
Chem
Biodivers
2015;
12:
1595-‐1634
[2] Zavala-‐Chavez
F,
Garcia-‐Mateos
R,
Soto-‐Hernandez
M,
Kite
G.
Phytochemical
differences
between
Calia
secundiflora
(Leguminosae)
growing
at
two
sites
in
Mexico.
Z
Naturforsch,
J
Biosci
2006;
61:
155-‐159
[3] Rios
MY,
Estrada-‐Soto
S,
Flores-‐Morales
V,
Aguilar
MI.
Chemical
constituents
from
Flouren-‐
sia
resinosa
S.F.
Blake
(Asteraceae).
Biochem
Syst
Ecol
2013;
51:
240-‐242
[4] Trute
A,
Nahrstedt
A.
Separation
of
rosmarinic
acid
enantiomers
by
three
different
chro-‐
matographic
methods
and
the
determination
of
rosmarinic
acid
in
Hedera
helix.
Phyto-‐
chem
Anal
1996;
7:
204-‐208
[5] Shimura
H,
Matsuura
M,
Takada
N,
Koda
Y.
An
antifungal
compound
involved
in
symbiotic
germination
of
Cypripedium
macranthos
var.
rebunense
(Orchidaceae).
Phytochem
2007;
68:
1442-‐1447
[6] King-‐Díaz
B,
Granados-‐Pineda
J,
Bah
B,
Rivero-‐Cruz
JF,
Lotina-‐Hennsen
B.
Mexican
propolis
flavonoids
affect
photosynthesis
and
seedling
growth.
J
Photochem
Photobiol
B
2015;
151:
213-‐220
P699
Synthesis
of
cyclic
citrullinated
peptides
targeting
rheumatoid
arthritis
autoantibodies
based
on
sunflower
trypsin
inhibitors
Camilla
Eriksson1,
Sunithi
Gunasekera1,
Cátia
Cerqueira2,
Per-‐Johan
Jacobsson2,
Ulf
Görans-‐
son1
1Div.
Pharmacognosy,
Dept.
Medicinal
Chemistry,
BMC,
Uppsala
University,
Uppsala,
Sweden,
2
Rheuma-‐
tology
Unit,
Dept.
Medicine,
Karolinska
Institutet,
Karolinska
University
Hospital,
Stockholm,
Sweden
Anti-‐cyclic
citrullinated
peptide
antibodies
(ACPA)
are
a
hallmark
of
rheumatoid
arthritis
(RA)
and
a
widely
used
biomarker
for
early
diagnosis.
ACPA
target
the
non-‐coded
amino
acid
citrulline
(deiminated
arginine)
on
peptides
and
proteins
present
during
RA.
ACPA
are
found
in
the
major
subset
of
the
RA-‐population
(60-‐70%)
[1]
and
possess
high
specificity
(87-‐96%)
[2].
Their
role
in
disease
progression
is
not
clear,
however
they
are
suggested
to
be
important
for
RA
pathophysiology
and
it
is
hypothesized
that
they
may
present
a
new
target
in
the
de-‐
velopment
of
disease-‐modifying
anti-‐rheumatic
drugs.
Among
the
best
characterized
ACPA-‐
targets
are
citrullinated
α-‐enolase,
vimentin,
fibrinogen,
filaggrin
and
type-‐II
collagen.
In
the
present
work,
we
developed
a
set
of
linear
and
cyclic
citrullinated
peptides
derived
from
se-‐
lected
target
proteins
to
determine
ACPA
selectivity.
Cyclic
peptides
from
natural
origin
were
used
as
scaffolds
to
improve
the
stability
of
peptide
epitopes
under
biological
conditions.
Our
results
from
competitive
ELISA
show
that
peptides
derived
from
fibrinogen,
filaggrin
and
col-‐
lagen
neutralize
ACPA
in
vitro.
These
results
support
the
hypothesis
of
cyclic
stable
peptide
analogues
as
potential
ACPA-‐neutralizers
or
as
diagnostic
tools.
Keywords:
ACPA,
rheumatoid
arthritis,
citrullination,
SFTI-‐1
References:
[1] Cerqueira
FC,
Klareskog
L,
Jakobsson
PJ.
Neutralization
of
anticitrullinated
protein
anti-‐
bodies
in
rheumatoid
arthritis
-‐
a
way
to
go?
Basic
Clin
Pharmacol
Toxicol
2014;
114:
13-‐
17
[2] Coenen,
D.,
Verscueren,
P.,
Westhovens,
R.,
Bossuyt,
X.
Technical
and
diagnostic
perfor-‐
mance
of
6
assays
for
the
measurement
of
citrullinated
protein/peptide
antibodies
in
the
diagnosis
of
rheumatoid
arthritis.
Clin
Chem
2007;
53:
498-‐504
P700
Silymarin
from
Silybum
marianum
–
new
approaches
to
separa-‐
tion
and
derivatization
David
Biedermann1,
Alena
Křenková1,
Kateřina
Valentová1,
Vladimír
Křen1
1
Institute
of
Microbiology,
Laboratory
of
Biotransformation,
Czech
Academy
of
Sciences,
Vídeňská
1083,
142 20 Prague, Czech Republic
Milk
thistle
(Silybum
marianum)
has
been
used
medicinally
from
the
14th
century
[1].
Today
it
is
a
source
of
silymarin
–
extract
of
its
fruits
–
which
is
used
as
nutraceutical
and
in
the
treatment
of
the
liver
problems.
Despite
high
commercial
value
and
large
amount
produced,
until
recently
preparative
separation
of
the
silymarin
component
has
not
been
achieved.
Con-‐
sequently,
extensive
research
of
its
biological
activity
was
not
possible.
Silymarin
was
separated
by
the
combination
of
Sephadex
LH-‐20
chromatography,
kinetic
res-‐
olution
and
crystallization
[2,3].
Silymarin
flavonolignans
were
all
obtained
in
gram
quantities
and
high
purity.
The
derivatization
was
mainly
carried
out
on
silychristin.[4]
The
toxicity
was
assayed
against
several
cell
lines
of
different
histological
origin,
the
cytotoxicity
is
mostly
neg-‐
ligible.
Radical
scavenging
activity
and
lipoperoxidation
inhibition
is
very
high
(eg.
anhydrosi-‐
lychristin
lipoperoxidation
inhibition
IC50=4.12
µM).
This
is
in
agreement
with
observed
low
redox
potentials
(dehydrosilychristin
0.39
V)
[4].
Supported
by
the
CSF
(project
No.
15-‐03037S),
MSMT
CZ
(project
LD
15080,
LD15081).
Keywords: Milk thistle, silymarin, silybin, silychristin, cytotoxicity, lipoperoxidation inhibi-‐
tion
References:
[1] Biedermann
D,
Vavrikova
E,
Cvak
L,
Kren
V.
Chemistry
of
silybin.
Nat
Prod
Rep
2014;
31:
1138-‐1157.
[2] Gazak
R,
Marhol
P,
Purchartova
K,
Monti
D,
Biedermann
D,
Riva
S,
Cvak
L,
Kren
V.
Large-‐
scale
separation
of
silybin
diastereoisomers
using
lipases.
Process
Biochem
2010;
45:
1657-‐1663.
[3] Krenek
K,
Marhol
P,
Peikerova
Z,
Kren
V,
Biedermann
D.
Preparatory
separation
of
the
si-‐
lymarin
flavonolignans
by
Sephadex
LH-‐20
gel.
Food
Res
Intl
2014;
65:
115-‐120.
[4] Pyszkova
M,
Biler
M,
Biedermann
D,
Valentova
K,
Kuzma
M,
Vrba
J,
Ulrichova
J,
Sokolova
R,
Mojovic
M,
Popovic-‐Bijelic
A,
Kubala
M,
Trouillas
P,
Kren
V,
Vacek
J.
Flavonolignan
2,3-‐
dehydroderivatives:
Preparation,
antiradical
and
cytoprotective
activity.
Free
Radic
Biol
Med
2016;
90:
114-‐125.
P701
Boosting
the
antifungal
drug
discovery
by
halogenating
plant
ex-‐
tracts
to
obtain
bioactive
‘unnatural’
natural
products
Davide
Righi,
Alice
Mainetti,
Quentin-‐Favre
Godal,
Laurence
Marcourt,
Jean-‐Luc
Wolfender,
Emerson
F.
Queiroz
School
of
Pharmaceutical
Sciences,
EPGL,
University
of
Geneva,
University
of
Lausanne,
30
Quai
Ansermet
CH-‐1211
Geneva
4,
Switzerland
It
is
estimated
that
20
percent
of
all
pharmaceutical
small
molecule
drugs
are
halogenated.1
Carbon–halogen
bond
lead
to
a
wide
range
of
effects,
including
an
increase
in
thermal
and
oxidative
stability
and
increased
biological
membrane
permeability
[1].
Furthermore
in
drug
discovery,
natural
products
(NPs)
represent
biologically
valuable
scaffolds
and
might
show
improved
bioactivities
or
bioavailability
when
halogenated.
Previous
works
have
demon-‐
strated
the
potential
of
chemical
halogenation
on
plant
extracts
to
obtain
bioactive
com-‐
pounds
[2].
In
this
context,
a
strategy
for
the
generic
halogenation
of
plant
extracts
has
been
developed
to
build
up
libraries
of
original
halogenated
NPs
in
particular
for
the
search
of
new
antifungal
agents.
First
the
halogenation
reaction
was
performed
with
a
series
of
NP
stand-‐
ards
from
different
chemical
classes.
Once
the
reaction
was
successfully
achieved
different
plant
extracts
have
been
submitted
to
the
same
generic
halogenation
procedure
(Br,
I,
Cl)
[3].
This
procedure
generated
significant
modification
of
the
metabolites
profiles
in
all
extracts
tested.
To
localize
the
active
compounds,
biological
profiling
for
antifungal
activity
was
per-‐
formed
using
at-‐line
HPLC-‐microfractionation
in
96-‐well
plates
and
subsequent
bioautog-‐
raphy
against
Candida
albicans
[4].
The
analytical
HPLC-‐PDA
conditions
were
geometrically
transferred
to
preparative
Flash
chromatography
column
(Flash-‐UV)
for
efficient
targeted
isolation.
Using
this
approach
a
series
of
halogenated
NP
analogues
possessing
interesting
antifungal
activity
has
been
identified.
80
60
40
20
0
0 10 20 30 40 50 60 min
Inactive Halogenation
Antifungal activity detected after HPLC
Plant
extract Reaction microfractionation and biological assay Active
Halogenated
NPs
Keywords: halogenation, chemical engineering, antifungal compounds, drug discovery
References:
[1] N.Herrera-‐Rodriguez L, Khan F, T.Robins K, Meyer HP. Perspectives on biotechnological
halogenation. Chem Today 2011; 29: 4
[2] Mendez L, O.Salazar M, Ramallo IA, Furlan LER. Brominated extracts as source of bioactive
compounds. ACS Comb Sci. 2011; 13: 200-‐204
[3] Bernini R, Pasqualetti M, Provenzano G, Tempesta S. Ecofriendly synthesis of halogenated
flavonoids and evaluation of their antifungal activity. New J Chem 2015; 39: 2980-‐2987
[4] Favre-‐Godal Q, Dorsaz S, Queiroz EF, Conan C, Marcourt L, Wardojo BP, Voinesco F,
Buchwalder A, Gindro K, Sanglard D, Wolfender JL. Comprehensive approach for the de-‐
tection of antifungal compounds using a susceptible strain of Candida albicans and con-‐
firmation
of
in
vivo
activity
with
the
Galleria
mellonella
model.
Phytochemistry
2014;
105:
68-‐78
P702
Sarqaquinoic
acid
and
related
synthetic
naphthoquinones
inhibit
the
function
of
Hsp90
Maynard
Chiwakata1,
Jo-‐Anne
de
la
Mare2,
Adrienne
Edkins2,
Denzil
R.
Beukes3
1
Faculty
of
Pharmacy,
Rhodes
University,
Grahamstown,
6140,
South
Africa,
2
Department
of
Biochemis-‐
try
and
Microbiology,
Rhodes
University,
Grahamstown,
6140,
South
Africa,
3
School
of
Pharmacy,
Univer-‐
sity
of
the
Western
Cape,
Bellville,
7535,
South
Africa
Heat
shock
protein
90
(Hsp90)
is
of
critical
importance
in
the
proper
folding
of
numerous
proteins,
including
those
involved
in
cancer.
Consequently,
there
is
significant
interest
in
the
discovery
and
development
of
Hsp90
inhibitors
as
anticancer
drugs.
In
this
study,
we
investi-‐
gated
the
ability
of
sargaquinoic
acid
(SQA)
and
selected
naphthoquinone
derivatives
to
inhib-‐
it
Hsp90
function.
SQA
was
isolated
and
purified
from
Sargassum
incisifolium
while
the
naph-‐
thoquinones
were
synthesised
via
a
straightforward
sequence
incorporating
a
Diels-‐Alder
reaction
between
benzoquinone
derivatives
and
myrcene
followed
by
coupling
with
substi-‐
tuted
alkyl
or
arylamines.
Hsp90
inhibition
was
assessed
by
a
client
protein
degradation
as-‐
say.
At
a
concentration
of
1
µM,
SQA
showed
almost
complete
inhibition
of
Hsp90
but
only
moderate
antiproliferative
effects
(IC50
658
µM)
against
a
Hs578T
breast
cancer
carcinoma
cell
line.
Interestingly,
the
most
potent
synthetic
aminonaphthoquinone
inhibited
Hsp90
function
by
50%
at
a
concentration
of
1
µM
but
showed
much
improved
activity
against
the
Hs578T
cell
line
(IC50
0.32
µM).
Furthermore,
unlike
geldanamycin,
none
of
the
compounds
tested
upregulates
Hsp70
suggesting
that
these
compounds
may
bind
to
the
C-‐terminal
end
of
Hsp90.
Keywords: Hsp90, sargaquinoic acid, naphthoquinone
P703
Assessment
of
the
bactericidal
effect
of
green
synthesized
silver
nanoparticles
against
a
panel
of
infectious
microorganisms
Edith
Antunes1,
Mokone
Mmola2,
3,
Mervin
Meyer2,
Denzil
R.
Beukes3
1
Department
of
Chemistry,
University
of
the
Western
Cape,
Robert
Sobukwe
Rd,
Bellville,
7535,
Cape
Town,
South
Africa,
2
Department
of
Biotechnology,
University
of
the
Western
Cape,
Robert
Sobukwe
Rd,
Bellville,
7535,
Cape
Town,
South
Africa,
3
School
of
Pharmacy,
University
of
the
Western
Cape,
Robert
Sobukwe
Rd,
Bellville,
7535,
Cape
Town,
South
Africa.
The
emergence
of
multi-‐drug
resistant
microorganisms
poses
a
major
threat
to
human
life,
making
the
antibiotics
currently
in
use,
ineffective
[1].
There
is
therefore
a
need
for
the
devel-‐
opment
of
new
broad-‐spectrum
antibiotics.
Current
research
has
been
channelled
into
nano-‐
science
in
conjunction
with
drug
discovery,
in
the
search
for
effective
antibacterial
agents.
Nanoscience
is
the
study
of
the
remarkable
properties
a
material
exhibits
when
it
is
reduced
to
less
than
100
nm
in
size.
The
synthesis
of
nanoparticles
can
be
achieved
using
the
so
called
“bottom-‐up”
or
“top-‐down”
approaches.
However,
the
chemicals
employed
are
typically
toxic
or
harmful,
restricting
their
use
in
medical
applications.
There
is
therefore
a
need
for
the
use
of
eco-‐friendly,
nanoparticle
synthetic
methods
i.e.
“green
chemistry”
methods
[2,3].
The
synthesis
of
the
silver
and
gold
nanoparticles
(NPs)
used
in
this
study
were
carried
out
by
means
of
green
synthetic
methods,
using
an
aqueous
extract
from
a
South
African
endemic
brown
alga
Sargassum
incisifolium.
For
comparison,
commercially
available
brown
algal
fu-‐
coidans
were
also
used
to
synthesise
the
NPs.
The
NPs
were
characterised
using
UV-‐Vis
and
FT-‐IR
spectroscopy,
Transmission
electron
microscopy,
Dynamic
light
scattering,
zeta
poten-‐
tial
measurements
and
X-‐ray
diffraction.
The
rate
of
NP
formation
clearly
varied
with
the
type
of
reducing
agent
used
and
the
NPs
produced
varied
in
size
from
5
nm
to
as
much
as
66
nm
for
both
sets
of
NPs.
The
AgNPs
synthesised
using
the
Sargassum
aqueous
extracts
showed
excellent
antimicrobial
activity
against
five
pathogenic
microorganisms
including
A.
bau-‐
mannii,
K.
pneumoniae,
E.
faecalis,
S.
aureus,
and
C.
albicans,
while
the
AuNPs
were
found
to
be
much
less
effective.
The
cytotoxic
activity
of
these
NPs
was
also
assessed
against
three
cell
lines,
namely
MCF-‐7,
HT-‐29
and
MCF-‐12a.
The
AgNPs
were
found
to
be
toxic
to
both
the
HT-‐
29
and
MCF-‐7
cell
lines,
exhibiting
slightly
less
toxicity
to
the
MCF-‐12a
cells.
The
AuNPs
showed
lower
toxicity
levels.
Acknowledgements:
The
authors
gratefully
acknowledge
UWC,
NRF
CSUR
and
CPRR
grants
(South
Afri-‐
ca)
and
the
DST/National
Nanoscience
Postgraduate
Teaching
and
Training
Programme
(NNPTTP)
for
financial
support
Keywords: Green synthesis, fucoidans, Sargassum incisifolium, antibacterial activity, cytotoxi-‐
city
References:
[1] Morones
JR,
Elechiguerra
JL,
Camacho
A,
Holt
K,
Kouri
JB,
Ramírez
JT,
Yacaman
MJ.
The
bac-‐
tericidal
effect
of
silver
nanoparticles.
Nanotechnol
2005;
16:
2346-‐2353
[2] Sun
Q,
Cai
X,
Li
J,
Zheng
M,
Chen
Z,
Yu
C-‐P.
Green
synthesis
of
silver
nanoparticles
using
tea
leaf
extract
and
evaluation
of
their
stability
and
antibacterial
activity.
Colloids
Surf
A
2014;
444:
226-‐231
[3] Akhtar
MS,
Panwar
J,
Yun
Y.
Biogenic
synthesis
of
metallic
nanoparticles
by
plant
extracts.
ACS
Sustainable
Chem
Eng
2013;
1:
591-‐602
P704
Biomimetic
studies
towards
the
total
synthesis
of
highly
complex
araiosamines
Kévin
Cottet,
Mehdi
A.
Beniddir,
Adam
Skiredj,
Laurent
Evanno,
Erwan
Poupon
BioCIS,
Univ.
Paris-‐Sud,
CNRS,
LabEx
LERMIT,
Université
Paris-‐Saclay,
92296
Châtenay-‐Malabry,
France
Araiosamines
are
among
the
most
complex
indole
alkaloids
isolated
from
marine
environ-‐
ments
to
date
[1].
Indeed,
the
fascinating
structures
of
several
araiosamines
were
disclosed
in
2011
(see
the
structure
of
araiosamine
C)
from
a
marine
sponge
(Clathria
araiosa)
[2].
Their
structures
reveal
an
original
assembly
of
tryptophane-‐derived
reactive
building
blocks
and
guanidine.
Based
on
biosynthetic
considerations,
several
cascades
of
reactions
are
studied
involving
simple
indole-‐acetaldehydes
and
guanidine
itself
in
order
to
better
understand
how
such
molecular
architectures
can
arise
in
Nature
[3].
Metabolomic
tools
appear
appropriate
for
such
approaches.
Assemblies
of
the
starting
units
give
rise
to
interesting
guanidine
heter-‐
ocycles.
Keywords: Araiosamine, alkaloids, guanidine
References:
[1] Berlinck
RGS,
Burtoloso
AC,
Trindale-‐Silva
AE,
Romminger
S,
Morais
RP,
Bandeira
K,
Mizu-‐
mo
CM.
The
chemistry
and
biology
of
organic
guanidine
derivatives.
Nat
Prod
Rep
2010;
27:
1871−1907
[2] Wei
X,
Henriksen
NM,
Skalicky
JJ,
Harper
MK,
Cheatham
TE,
Ireland
CM,
Van
Wagoner
RM.
Araiosamines
A–D:
Tris-‐bromoindole
Cyclic
Guanidine
Alkaloids
from
the
Marine
Sponge
Clathria
(Thalysias)
araiosa.
J
Org
Chem
2011;
76:
5515–5523
[3] Genta-‐Jouve
G,
Cachet
N,
Holderith
S,
Obershänli
F,
Teyssié
JL,
Jeffree
R,
Al
Mourabit
A,
Thomas
OP.
New
insight
into
marine
alkaloid
metabolic
pathways:
revisiting
oroidin
bio-‐
synthesis.
ChemBioChem
2011;
12:
2298−2301
P705
Antimalarial
and
antimicrobial
activities
of
α-‐(2-‐piperidyl)-‐2-‐
aryl-‐4-‐quinolinemethanol
analogs
H.
M.
T.
Bandara
Herath,
H.
Ranjith
W.
Dharmaratne,
Melissa
Jacob,
Shabana
I.
Khan,
N.
P.
Dhammika
Nanayakkara
National
Center
for
Natural
Product
Research,
School
of
Pharmacy,
University
of
Mississippi,
University,
MS
38677
Quinine,
an
alkaloid
isolated
from
cinchona
tree,
has
been
used
for
the
treatment
of
malaria
for
more
than
two
centuries
[1].
During
the
last
century,
a
number
of
synthetic
analogs
of
qui-‐
nine
have
been
prepared
and
evaluated
for
antimalarial
activity
[2].
Mefloquine,
a
more
po-‐
tent
quinine
analog,
is
currently
used
for
the
treatment
and
prevention
of
malaria.
Although
quinine
and
mefloquine
have
weak
antimicrobial
activities,
their
2-‐aryl-‐analogs
have
shown
potent
activity
against
several
opportunistic
fungi
and
pathogenic
bacteria
[3].
In
order
to
develop
basic
structure-‐activity
relationships,
we
have
prepared
several
α-‐(2-‐
piperidyl)-‐2-‐aryl-‐4-‐quinolinemethanol
analogs
(1:
R1,
R2,
R3,
R4
=
H,
Cl,
or
CF3)
and
evaluated
them
for
in
vitro
antiplasmodial
and
antimicrobial
activities.
These
analogs
showed
potent
activity
against
P.
falciparum
W2
and
D6
strains
(IC50
6.5–30
ng/ml;
positive
control,
meflo-‐
quine,
IC50
=
20.0
ng/ml)
as
well
as
opportunistic
fungi,
Candida
albicans
(IC50
=
2.0–10.0
µg/ml;
positive
control,
amphotericin
B,
IC50
=
0.3
µg/ml),
Cryptococcus
neoformans
(IC50
=
0.30–0.70
µg/ml;
positive
control,
amphotericin
B,
IC50
=
0.3
µg/ml)
,
and
Aspergillus
fumiga-‐
tus
(IC50
=
0.30–3.5
µg/ml;
positive
control,
amphotericin
B,
IC50
=
0.9
µg/ml),
and
bacteria,
methicillin-‐resistant
Staphylococcus
aureus
(IC50
=
0.35–1.2
µg/ml;
positive
control,
ciproflox-‐
acin,
IC50
=
0.1
µg/ml)
and
Mycobacterium
intracellulare
(IC50
=
0.23–1.5
µg/ml;
positive
con-‐
trol,
ciprofloxacin,
IC50
=
0.4
µg/ml).
These
results
provide
a
basis
for
further
development
of
this
class
of
compounds
for
treatment
of
malarial
and
microbial
infections.
Keywords:
4-‐Quinolinemethanol,
antimalarial,
antimicrobial
References:
[1] Achan
J,
Talisuna
AO,
Erhart
A,
Yeka
A,
Tibenderana
JK,
Baliraine
FN,
Rosenthal
PJ,
D'Alessandro
U.
Quinine,
an
old
anti-‐malarial
drug
in
a
modern
world:
role
in
the
treatment
of
malaria.
Malar
J
2011;
10:
144
[2] Schmidt
LH,
Crosby
R,
Rasco
J,
Vaughan
D.
Antimalarial
activities
of
various
4-‐
quinolonemethanols
with
special
attention
to
WR-‐142,490
(mefloquine).
Antimicrob
Agents
Chemother
1978;
13:
1011-‐1030
[3] Kunin
CM,
Ellis
WY.
Antimicrobial
activities
of
mefloquine
and
a
series
of
related
com-‐
pounds.
Antimicrob
Agents
Chemother
2000;
44:
848-‐852
P706
Establishment
and
evaluation
of
processes
for
the
production
of
the
antiviral
and
cytostatic
cardenolide
glucoevatromonoside
Jennifer
Munkert1,
Marina
Santiago
Franco2,
Fernao
C.
Braga2,
Wolfgang
Kreis1,
Saulo
F.
An-‐
drade3,
Flaviano
Melo
Ottoni2,
Ricardo
José
Alves2,
Rodrigo
Maia
de
Pádua2
1
Department
of
Biology,
Friedrich-‐Alexander
Universität,
Erlangen-‐Nürnberg,
Germany;
2
Faculdade
de
Farmácia,
Departamento
de
Produtos,
Farmacêuticos,
Universidade
Federal
de
Minas
Gerais,
Belo
Hori-‐
zonte,
MG,
Brazil;
3
Departamento
de
Produção
de
Matéria-‐Prima,
Faculdade
de
Farmácia,
Universidade
Federal
do
Rio
Grande
do
Sul
(UFRGS),
Av.
Ipiranga,
2752,
Porto
Alegre,
Brazil
Cardenolides
(cardiac
glycosides)
are
a
group
of
natural
products
possessing
interesting
bio-‐
logical
activities.
For
decades,
cardiac
glycosides
have
been
used
for
treating
cardiac
insuffi-‐
ciency
in
humans,
as
they
bind
to
and
inhibit
Na+/K+-‐ATPase
in
the
myocard
resulting
in
an
increased
cardiac
output
by
increasing
the
force
of
contraction.
Besides
the
well-‐known
posi-‐
tive
effects
on
heart
activity,
new
therapeutic
targets
in
various
diseases
are
discussed.
More
recent
studies
reported
an
increased
susceptibility
of
cancer
cells
to
cardenolides
in
tumor
therapy
and
anti-‐cancer
treatment
[1,2].
Other
targets
are
viral
diseases,
such
as
Herpes
sim-‐
plex
virus
infection
(HSV).
A
very
strong
anti-‐herpes
effect
was
recently
attributed
to
the
car-‐
diac
glycoside
glucoevatromonoside
(GEV)
[3].
Although
cardiac
glycosides
have
a
great
po-‐
tential
for
anti-‐cancer
and
anti-‐virus
therapy,
they
are
still
isolated
from
plants
since
their
structural
complexity
impedes
their
chemical
synthesis.
GEV
is
currently
not
commercially
available.
For
the
previous
studies
GEV
was
isolated
from
Digitalis
lanata
in
a
time
consuming
process
[4].
Therefore
we
suggest
and
present
here
additional
more
straight-‐forward
ap-‐
proaches
to
obtain
the
bioactive
compound
GEV.
We
consider
and
compare
a
biotransfor-‐
mation
of
suitable
precursors
by
plant
suspension
cultures,
chemical
synthesis
of
GEV
and
a
molecular
biology
approach.
For
biotransformation
reaction
K1OHD
suspension
cells
from
D.
lanata
are
feed
with
synthesized
digitoxigenin-‐monodigitoxoside.
Final
recovering
by
bio-‐
transformation
of
purified
GEV
was
up
to
15
%.
However
the
developed
regioselective
catalyst
controlled
glycosylation
reaction
resulted
in
45
%
purified
GEV.
Regarding
the
molecular
ap-‐
proach,
we
established
a
set
of
recombinant
glycosyltransferases
belonging
to
the
Glycosyl-‐
transferase
Family
1
from
either
the
cardenolid
containing
plant
E.
crepidifolium
or
form
the
model
plant
A.
thaliana
that
can
be
used
in
in
vitro
or
in
vivo
enzyme
assays.
Acknowledgements:
EU
FP7
IRSES
(grant
295251),
CNPq
Keywords: Cardenolides, glucoevatromonoside, plant suspension culture, chemical synthesis,
glucosyltransferase
References:
[1] Prassas
I,
Diamandis
EP.
Novel
therapeutic
applications
of
cardiac
glycosides.
Nat
Rev
Drug
Discov
2008;
7:
926-‐935
[2] Newman
RA,
Yang
P,
Pawlus
AD,
Block
KI.
Cardiac
glycosides
as
novel
cancer
therapeutic
agents.
Mol
Interv
2008;
8:
36-‐49
[3] Bertol
JW,
Rigotto
C,
de
Pádua
RM,
Kreis
W,
Barardi
CR,
Braga
FC,
Simões
CM.
Antiherpes
activity
of
glucoevatromonoside,
a
cardenolide
isolated
from
a
Brazilian
cultivar
of
Digi-‐
talis
lanata.
Antiviral
Res
2011;
92:
73-‐80.
[4] Braga
FC,
Kreis
W,
Braga
de
Oliveira
A.
Isolation
of
cardenolides
from
a
Brazilian
cultivar
of
Digitalis
lanata
by
rotation
locular
counter-‐current
chromatography.
J
Chromatogr
A
1996;
756:
287-‐291
P707
Synthesis
and
characterization
99mTc-‐labebled
DTPA-‐
digitoxigenin
and
its
new
potential
in
imaging
techniques
for
the
diagnostic
and
identification
of
tumor
cells
Jennifer
Munkert1,
Eliza
Rocha
Gomes2,
Saulo
F.
Andrade3,
José
Dias
de
Souza
Filho4,
Lucas
L.
Marostica5,
Wolfgang
Kreis1,
Fernão
C.
Braga2,
Cláudia
M.
O.
Simões5, Valbert
Nascimento
Cardoso2,
Rodrigo
M.
Pádua2*,
André
Luís
Branco
de
Barros2*
1
Department
of
Biology,
Friedrich-‐Alexander
Universität,
Erlangen-‐Nürnberg,
Germany;
2
Faculdade
de
Farmácia,
Departamento
de
Produtos,
Farmacêuticos,
Universidade
Federal
de
Minas
Gerais,
Belo
Hori-‐
zonte,
MG,
Brazil;
3
Departamento
de
Produção
de
Matéria-‐Prima,
Faculdade
de
Farmácia,
Universidade
Federal
do
Rio
Grande
do
Sul
(UFRGS),
Av.
Ipiranga,
2752,
Porto
Alegre,
Brazil;
4
Departamento
de
Química,
Universidade
Federal
de
Minas
Gerais,
Belo
Horizonte,
MG,
Brazil;
5
Departamento
de
Ciências
Farmacêuticas,
Centro
de
Ciências
da
Saúde,
Universidade
Federal
de
Santa
Catarina,
Florianópolis,
SC,
Brazil
In
recent
years
cardenolides
and
cardiac
glycosides,
have
presented
novel
therapeutic
appli-‐
cations,
as
antiviral
and
anticancer
drugs
[1].
Digitoxigenin
is
known
to
bind
and
inhibit
Na+/K+-‐ATPase
and
furthermore
tumor
cells
are
more
susceptible
to
bind
cardenolides,
as
they
either
show
a
higher
expression
rate
of
the
Na+/K+-‐ATPase
protein
or
a
higher
affinity
towards
the
binding
of
cardenolides
[2].
As
cancer
imaging
techniques
using
radiotracers
tar-‐
geted
to
specific
receptors
have
yielded
successful
results,
the
utility
of
such
approaches
for
developing
specific
radiopharmaceuticals
was
demonstrated
[3].
Technetium-‐99m
(99mTc)
has
mostly
been
used
to
label
radiopharmaceuticals,
due
to
its
suitable
physical
and
chemical
characteristics
and
inexpensive
isotope
cost
[3].
Diethylene
triamine
pentaacetic
acid
(DTPA)
is
an
attractive
bifunctional
chelating
ligand
used
to
prepare
99mTc-‐labeled
complexes.
There-‐
fore
the
radiolabeling
of
a
complex
of
Digitoxigenin
and
DTPA
with
99mTc
was
tried
as
a
new
approach
for
the
use
in
imaging
technique.
By
radiolabeling
the
complex
of
Digitoxigenin
and
DTPA
the
biodistribution
can
be
followed
and
therefore
this
complex
might
be
useful
in
diag-‐
nostic
and
identification
of
tumor
cells.
We
here
present
the
results
of
the
complex
synthesis,
as
well
as
the
characterization
of
the
radiolabeled
compound
including
stability
tests,
blood
clearance,
biodistribution
studies
in
healthy
mice
and
the
specific
binding
of
the
complex
to
A
549
lung
cancer
cells.
Acknowledgements: EU FP7 IRSES (grant 295251), CNPq. * Authors contributed equally to work
Keywords: Cardenolides, digitoxigenin, technetium-‐99m (99mTc), diagnosis agents
References:
[1] Prassas
I,
Diamandis
EP.
Novel
therapeutic
applications
of
cardiac
glycosides.
Nat
Rev
Drug
Discov.
2008;
7:
926-‐935.
[2] Newman
RA,
Yang
P,
Pawlus
AD,
Block
KI.
Cardiac
glycosides
as
novel
cancer
therapeutic
agents.
Mol
Interv
2008;
8:
36-‐40.
[3] de
Barros
ABL,
Mota
LG,Ferreira
CA,Corrêa
NCR,
de
Góes
AM,
Oliveira
MC,
Cardoso
VN.
99mTc-‐labeled
bombesin
analog
for
breast
cancer
identification.
J
Radioanal
Nucl
Chem
2013;
295:2083-‐2090.
P708
Four
new
diterpenoid
alkaloids
from
Aconitum
japonicum
Koji
Wada1,
Keiko
Takeda1,
Machiko
Haraguchi1,
Yuki
Abe1,
Natsumi
Kuwahara1,
Shota
Suzu-‐
ki1,
Ayaka
Terui1,
Takumi
Masaka1,
Naoko
Munakata1,
Mariko
Uchida1,
Masashi
Nunokawa1,
Hiroshi
Yamashita1,
Masuo
Goto2,
Kuo-‐Hsiung
Lee2,3
1
Medicinal
Chemistry,
School
of
Pharmacy,
Hokkaido
Pharmaceutical
University,
7-‐15-‐4-‐1,
Maeda,
Teine,
Sapporo
006-‐8590,
Japan,
2
Natural
Products
Research
Laboratories,
UNC
Eshelman
School
of
Pharmacy,
University
of
North
Carolina,
Chapel
Hill,
NC
27599
7568,
USA,
3
Chinese
Medicine
Research
and
Devel-‐
opment
Center,
China
Medical
University
and
Hospital,
Taichung,
Taiwan
The
constituents
of
Aconitum
japonicum
THUNB
(Ranunculaceae)
were
investigated
many
years
ago,
and
the
isolation
and
structure
elucidation
of
twenty-‐three
diterpenoid
alkaloids
from
rhizomes
of
Aconitum
species
have
been
reported
[1-‐7].
The
investigation
on
the
con-‐
stituents
of
this
plant
have
now
resulted
in
the
isolation
of
four
new
C19-‐diterpenoid
alkaloids,
14-‐anisoyl-‐N-‐deethylaconine
(1),
N-‐deethylaljesaconitine
A
(2),
14-‐anisoyllasianine
(3),
and
N-‐deethylnevadensine
(4),
together
with
fifteen
known
C19-‐diterpenoid
alkaloids.
Alkaloids
were
isolated
using
standard
column
liquid
chromatography
and
structures
assigned
by
high
resolution
mass
spectrometry
combined
with
C13
and
1
and
2D
proton
NMR
spectroscopy.
Two
of
the
new
C19-‐diterpenoid
alkaloids
(2,
4)
and
three
of
the
known
diterpenoid
alkaloids
(aconine
5,
virescenine
6,
ryosenamine
7)
were
evaluated
for
cytotoxic
activity
against
four
human
tumor
cell
lines
[lung
carcinoma
(A549),
triple-‐negative
breast
cancer
(estrogen
and
progesterone
receptors-‐negative/HER2-‐negative)
(MDA-‐MB-‐231),
nasopharyngeal
(KB),
and
multidrug
resistant
KB
subline
expressing
P-‐glycoprotein
(KB-‐VIN)].
However,
the
five
diterpenoid
alkaloids
(2,
4-‐7)
were
inactive
(IC50
>
40 µM)
against
four
human
tumor
cell
lines
(A549,
MDA-‐MB-‐231,
KB,
and
KB-‐VIN).
The
presentation
describes
the
isolation
of
four
new
aconitine-‐
or
lycoctonine-‐type
C19-‐diterpenoid
alkaloids
(1-‐4)
from
the
rhizomes
of
A.
japonicum.
The
structures
of
1-‐4
were
elucidated
from
1D
and
2D
NMR
spectroscopic
data.
The
structure
of
three
new
alkaloids,
14-‐anisoyl-‐N-‐deethylaconine
(1),
N-‐
deethylaljesaconitine
A
(2),
and
N-‐deethylnevadensine
(4),
was
unusual
N-‐deethyl
C19-‐
diterpenoid
alkaloids.
14-‐Anisoyllasianine
(3)
was
the
fourth
natural
C19-‐diterpenoid
alkaloid
possessing
the
amino
group
at
C-‐8.
Lipoaconitine,
lipomesaconitine,
aconine
(5),
15α-‐
hydroxyneoline,
isotalatizidine,
nevadenine,
talatisamine,
virescenine
(6),
nevadensine,
ry-‐
osenamine
(7),
and
dehydrolucidusculine
were
isolated
the
first
time
from
this
plant.
OH
OCH3 OCH3 OCH3
OCH3 OR3 OH OH OH
H H H
H H H 3C H
R2
R2 N N N
OH O
HO R1 OH OH
H H H
R1 OH
OCH3
H 3CO H 3CO H 3CO
1: R1 = OH, R 2 = H, R 3 = As 4: R1 = OH, R 2 = H 6
2: R1 = OCH3, R 2 = H, R 3 = As
3: R1 = NH 2, R 2 = CH2CH3, R 3 = As
5: R1 = OH, R 2 = CH2CH3, R 3 = H
As = COC6H 4OCH3 (p)
BzO
OH
N OH
7
Keywords:
Aconitum
japonicum,
C19-‐diterpenoid
alkaloid,
14-‐anisoyl-‐N-‐deethylaconine,
N-‐
deethylaljesaconitine
A,
14-‐anisoyllasianine,
N-‐deethylnevadensine
References:
[1] Bando
H,
Kanaiwa
Y,
Wada
K,
Mori
T,
Amiya
T.
Structure
of
deoxyjesaconitine.
A
new
diterpene
alkaloid
from
Aconitum
subcuneatum
NAKAI.
Heterocycles
1981;
16:
1723-‐
1726
[2] Mori
T,
Bando
H,
Kanaiwa
Y,
Wada
K,
Amiya
T.
Studies
on
the
constituents
of
Aconitum
Species.
II.
Structure
of
deoxyjesaconitine.
Chem
Pharm
Bull
1983;
31:2884-‐2886
[3] Wada
K,
Bando
H,
Mori
T,
Wada
R,
Kanaiwa
Y,
Amiya
T.
Studies
on
the
constituents
of
Ac-‐
onitum
Species.
III.
On
the
components
of
Aconitum
subcuneatum
NAKAI.
Chem
Pharm
Bull
1985;
33:
3658-‐3661
[4] Bando
H,
Wada
K,
Watanabe
M,
Mori
T,
Amiya
T.
Studies
on
the
constituents
of
Aconitum
Species.
IV.
On
the
components
of
Aconitum
japonicum
THUNB.
Chem
Pharm
Bull
1985;
33:
4717-‐4722
[5] Bando
H,
Wada
K,
Amiya
T,
Fujimoto
Y,
Kobayashi,
K.
Structures
of
secojesaconitine
and
subdesculine,
two
new
diterpenoid
alkaloids
from
Aconitum
japonicum
THUNB.
Chem
Pharm
Bull
1988;
36:
1604-‐1606
[6] Bando
H,
Wada
K,
Amiya
T,
Fujimoto
Y,
Kobayashi
K.
Studies
on
the
constituents
of
Aconi-‐
tum
Species.
VII.
On
the
components
of
Aconitum
japonicum
THUNB.
Heterocycles
1988;
27:
2167-‐2174
[7] Wada
K.
Studies
on
structural
elucidation
of
Aconitum
diterpenoid
alkaloid
by
LC-‐APCI-‐
MS
and
effects
of
Aconitum
diterpenoid
alkaloid
on
cutaneous
blood
flow.
Yakugaku
Zas-‐
shi
2002;
122:
929-‐956
P709
Phytotoxic
constituents
of
Diaporthe
eres
and
synthesis
of
ana-‐
logs
Kumudini
M.
Meepagala1,
Natascha
Techen2,
Robert
D.
Johnson1,
Stephen
O.
Duke1
1
USDA-‐ARS,
NPURU,
P.
O.
Box
1848,
University,
MS
38677,
U.S.A,
2
National
Center
for
Natural
Product
Research, University, MS 38677, USA
Plant
pathogenic
fungi
produce
secondary
metabolites
that
are
phytotoxic
to
the
host
plant.
These
metabolites
can
be
used
as
bioherbicides
or
lead
molecules
or
templates
in
developing
agrochemicals
[1].From
an
infected
leaf
of
Hedera
helix
(English
Ivy)
that
showed
necrosis,
a
fungus
was
isolated
and
identified
as
Diaporthe
eres
by
molecular
techniques.
The
ITS1-‐5.8S-‐
ITS2
genomic
region
(ITS)
was
amplified
from
genomic
DNA
using
the
forward
primer
ITS1
(5´-‐
TCCGTAGGTGAACCTGCGG-‐3´)
and
the
reverse
primer
ITS4
(5´-‐
TCCTCCGCTTATTGA-‐
TATGC-‐3´)
[2].
The
primers
cylh3f
(5´-‐
AGGTCCACTGGTGGCAAG-‐3´)
[3]
and
H3-‐1b
(5′-‐
GCGGGCGAGCTGGATGTCCTT-‐3′)
[4]
were
used
to
amplify
part
of
the
histone
H3
(HIS)
gene,
and
the
primers
T1
(5′-‐
AACATGCGTGAGATTGTAAGT-‐3′)
[5]
and
Bt-‐2b
(5′-‐
ACCCTCAGTG-‐
TAGTGACCCTTGGC-‐3′)4
to
amplify
part
of
the
beta-‐tubulin
gene
(TUB).
This
fungus
was
grown
in
Czapek
Dox
broth
culture
medium
for
two
weeks.
The
ethyl
acetate
extract
of
the
liquid
culture
medium
showed
phytotoxic
activity
against
monocots
(Agrostis
stolinifera)
and
dicots
(Lactuca
sativa)
with
90%
and
80%
germination
inhibition
respectively.
The
two
known
metabolites
were
isolated
via
bioassay
guided
isolation
and
identified
by
NMR
spec-‐
troscopy.
This
is
the
first
report
of
isolation
of
phytotoxins
from
D.eres.
One
of
the
phytotoxic
constituents
was
identified
as
8-‐hydroxy-‐3,
7
-‐dimethylisochroman-‐1-‐one.
We
have
synthe-‐
sized
this
phytotoxin
as
well
as
some
analogs
that
showed
phytotoxic
activity
in
seed
germi-‐
nation
bioassays
and
Lemna
pausicostata
(duckweed)
growth
bioassay.
The
IC50
values
of
these
compounds
ranged
from
82
µM
to
288
µm
in
L.
pausicostata
bioassay.
Some
analogs
showed
hormesis
in
L.
pausicostata
growth
bioassay.
Acknowledgements:
Eric
Briscoe,
Brandon
Clausen
and
Bradford
Gilbreath
are
acknowledged
for
tech-‐
nical
assistance
Keywords: Diaporthe eres, Hedera helix, phytotoxins, isochroman
References:
[1] Cimmino
A,
Masi
M,
Evidente
M,
Superchi
S,
Evidente
A.
Fungal
phytotoxins
with
potential
herbicidal
activity:
chemical
and
biological
characterization.
Nat
Prod
Rep
2015;
32:
1629-‐1653
[2] White,
TJ,
Burns
T,
Lee
S,
Taylor
J.
Amplification
and
direct
sequencing
of
fungal
ribosomal
RNA
genes
for
phylogenetics.
PCR
Protocols:
A
guide
to
methods
and
applications.
1990;
315-‐322
[3] Crous
PW,
Groenewald
JZ,
Risède
J-‐M,
Simoneau
P,
Hyde
KD.
Calonectria
specis
and
their
cylindrocladium
anamorphs:
Species
with
clavate
vesicles.
Stud
Mycol
2006;
55:
213-‐226
[4] Glass
NL
and
Donaldson
GC.
Development
of
primer
sets
designed
for
use
with
the
PCR
to
amplify
conserved
genes
from
filamentous
ascomycetes.
Applied
Environmental
Microbi-‐
ology.
1995;
61:
1323-‐1330
[5] O’Donnell
K
and
Cigelnik
E.
Two
divergent
intragenomic
rDNA
ITS2
types
within
a
monophletic
lineage
of
the
fungus
fusarium
are
non
orthologous.
Molecular
phylogenetic
Evolution.
1997;
7:
103-‐116
P710
A
flavone
and
cytotoxic
activity
of
sesquiterpenoids
from
the
res-‐
inous
exudates
of
cushion
bush
(Leucophyta
brownii)
Katrine
T.
Jensen1,
Mette
G.
Hyldgaard1,
Stig
Purup2,
Xavier
Fretté1,
Lars
P.
Christensen1
1
Department
of
Chemical
Engineering,
Biotechnology
and
Environmental
Technology,
University
of
Southern
Denmark,
Campusvej
55,
5230
Odense
M,
Denmark,
2
Department
of
Animal
Science,
Aarhus
University,
Blichers
Allé
20,
8830
Tjele,
Denmark
A
reinvestigation
of
the
resinous
exudates
of
cushion
bush
(Leucophyta
brownii
Cass.,
Aster-‐
aceae)
resulted
in
the
isolation
of
the
8,12-‐guaianolides
1−4,
the
xanthanolide
tomentosin
(5),
and
the
1,10-‐seco-‐eudesmane
leucophytalin
C
(6)
previously
isolated
from
this
plant
[1],
as
well
as
the
flavone
7,4’-‐dihydroxy-‐5,8-‐dimethoxyflavone
(7).
The
flavone
7
has
previously
been
synthesized
[2]
but
to
the
best
of
our
knowledge
this
is
its
first
report
from
natural
sources.
The
flavone
7
was
identified
by
1D
and
2D
NMR
techniques,
UV
and
LC-‐MS/MS.
Ses-‐
quiterpene
lactones
containing
an
α,
β-‐unsaturated
γ-‐lactone
moiety
are
known
for
their
anti-‐
inflammatory
and
cytotoxic
activity
due
to
reactions
with
sulfhydryl
groups
of
functional
pro-‐
teins
via
a
Michael-‐type
reaction
[1].
Compounds
1−6
were
investigated
for
their
cytotoxic
activity
towards
human
breast
cancer
(MCF-‐7)
and
colon
cancer
(HT-‐29)
cells
and
their
abil-‐
ity
to
induce
apoptosis.
Compounds
1−4
reduced
proliferation
of
HT-‐29
and
MCF-‐7
cells
be-‐
tween
60−90%
at
a
concentration
of
18.9,
16.3,
20.2
and
17.2
µM,
respectively,
and
with
IC50
values
<
10
µM.
Tomentosin
(5)
showed
less
cytotoxicity
with
an
IC50
value
of
approximately
40
µM
for
both
cell
lines.
As
expected
leucophytalin
C
(6)
with
the
absence
of
an
α,
β-‐
unsaturated
γ-‐lactone
moiety
did
not
show
any
significant
cytotoxicity
at
a
concentration
of
79.4
µM.
No
activation
of
caspase-‐3,
-‐7
and
-‐8
were
observed
in
the
tested
cancer
cell
lines.
This
was
also
confirmed
by
gene
expression
studies,
indicating
that
the
observed
cytotoxic
effect
of
compounds
1−5
was
not
due
to
an
apoptosis
initiated
mechanism.
Acknowledgements:
A.
K.
Nielsen
and
K.
B.
Poulsen
are
acknowledged
for
technical
assistance
Keywords: Leucophyta brownii, 7,4’-‐dihydroxy-‐5,8-‐dimethoxyflavone, 8,12-‐guaianolides, cy-‐
totoxicity, apoptosis
References:
[1] Hyldgaard
MG,
Purup
S,
Bond
AD,
Fretté
XC,
Qu
H,
Jensen
KT,
Christensen
LP.
Guaianolides
and
a
seco-‐eudesmane
from
the
resinous
exudates
of
cushion
bush
(Leucophyta
brownii)
and
evaluation
of
their
cytostatic
and
anti-‐inflammatory
activity.
J
Nat
Prod
2015;
78:
1877-‐1885
[2] Gupta
SR,
Seshadri
TR,
Sharma
CS,
Sharma
ND.
Chemical
investigation
of
Dikamali
gum:
isolation
of
a
new
flavone,
4'-‐hydroxywogonin.
Indian
J
Chem
1975;
13:
785-‐788
P711
Biomimetic
assembly
of
leucoridine
A
Sarah Benayad, Mehdi A. Beniddir, Laurent Evanno, Erwan Poupon
BioCIS, Univ. Paris-‐Sud, CNRS, LabEx LERMIT, Université Paris-‐Saclay, 92296 Châtenay-‐Malabry, France
Leucoridine
A
is
a
complex
bis-‐indole
alkaloid
isolated
in
2010
from
Leuconotis
griffithii
(Apocynaceae)
showing
moderate
cytotoxicity
towards
human
KB
cells
(IC50
=
1
μM)
[1].
The
three-‐step
biomimetic
assembly
of
leucoridine
A
is
described
[2].
The
hemisynthetic
route
provides
suitable
conditions
toward
the
central
3-‐spiro-‐1,
2,
3,
4-‐dehydropiperidine
ring
con-‐
necting
the
two
subunits
of
this
highly
congested
structure.
The
biomimetic
assembly
by
an
imino-‐Rauhut-‐Currier
reaction
affords
the
natural
(S)-‐diastereomer
of
leucoridine
A
as
the
sole
product.
The
sequence,
repeated
in
a
one-‐pot
procedure,
supports
a
non-‐enzymatic
pathway
for
the
assembly
of
this
complex
alkaloid
and
known
related
alkaloids.
Keywords:
Leucoridine
A,
Biomimetic
synthesis,
Alkaloid
References:
[1] Gan
CY,
EtOH
T,
Hayashi
M,
Komiyama
K,
Kam
TS.
Leucoridines
A-‐D,
cytotoxic
Strychnos-‐
Strychnos
bisindole
alkaloids
from
Leuconotis.
J
Nat
Prod
2010;
73:
1107-‐1111
[2] Benayad
S,
Beniddir
MA,
Evanno
L,
Poupon
E.
Biomimetic
assembly
of
Leucoridine
A.
Eur
J
Org
Chem
2015;
1894-‐1898
P712
Preakuammicine:
A
long
awaited
missing
link
in
the
biosynthesis
of
monoterpene
indole
alkaloids
Sarah Benayad, Kadiria Ahamada, Guy Lewin, Laurent Evanno, Erwan Poupon
BioCIS, Univ. Paris-‐Sud, CNRS, LabEx LERMIT, Université Paris-‐Saclay, 92296 Châtenay-‐Malabry, France
Preakuammicine
is
considered
as
a
key
intermediate
in
the
biosynthesis
of
monoterpene
in-‐
dole
alkaloids
since
the
1970's.
Indeed,
preakuammicine
plays
a
crucial
role
in
the
biosynthe-‐
sis
of
strychnan,
type
II,
type
III
monoterpene
indole
alkaloids.
According
to
the
seminal
hy-‐
pothesis,
preakuammicine
would
originate
from
a
complex
rearrangement
of
the
akuammilan
skeleton
(such
as
rhazimol)
but
neither
the
structure
of
preakuammicine,
nor
the
direct
con-‐
version
from
akuammillan
alkaloids
have
been
described.
[1]
Given
the
piecemeal
spectral
description
of
the
molecule,
[2]
questions
about
its
structure
and
in
particular
a
non
matching
C-‐16
stereochemistry
between
different
monoterpene
indole
alkaloid
series
[ex:
rhazimol
(16R),
stemmadenine
(16S)].
The
conversion
of
akuammilan-‐type
into
strychnan-‐type
alka-‐
loids
was
therefore
studied,
with
the
aim
of
characterizing
this
“missing
link”
for
the
first
time.
The
unprecedented
biomimetic
transformation
of
the
akuammilan
framework
into
acetyl-‐
16R-‐preakuammicine
was
performed
in
buffered
conditions.
Retention
of
the
C16
configura-‐
tion
was
observed
giving
mechanistic
insight
supporting
a
sigmatropic
pathway.
This
work
affords
the
first
spectral
data
for
this
never-‐described
structure.
Importantly
the
16R
stereo-‐
chemistry
of
preakuammicine
as
obtained,
does
not
match
the
16S
stereochemistry
of
stem-‐
madenine.
These
results
lead
for
the
first
time
to
the
consideration
of
both
the
16R
and
the
16S
diasteroisomers
of
preakuammicine
as
biosynthetic
precursors
intervening
in
two
co-‐
existing
routes.
Keywords: Preakuammicine, biosynthesis, alkaloids, sigmatropy [1, 5]
References:
[1] O’Connor
SE,
Maresh
J.
Chemistry
and
biology
of
monoterpene
indole
alkaloid
biosynthe-‐
sis.
Nat
Prod
Rep
2006;
23:
532-‐547
[2] Scott
AI,
Qureshi
AA.
Biogenesis
of
Strychnos,
Aspidosperma,
and
Iboga
alkaloids.
Struc-‐
ture
and
reactions
of
preakuammicine.
J
Am
Chem
Soc
1969;
91:
5874-‐5876
[3] Benayad
S,
Ahamada
K,
Lewin
G,
Evanno
L,
Poupon
E.
Preakuammicine:
A
long-‐awaited
missing
link
in
the
biosynthesis
of
monoterpene
indole
alkaloids.
Eur
J
Org
Chem
2016;
1494-‐1499
P713
Cytotoxic
potential
of
naturally
occurring
isoquinoline
alkaloids
possessing
different
structural
types
Cahlíková
Lucie1,
Doskočil
Ivo2,
Chlebek
Jakub1,
Hošťálková
Anna1,
Havelek
Radim3,
Šafratová
Marcela1
1
ADINACO
Research
Group,
Department
of
Pharmaceutical
Botany
and
Ecology,
Faculty
of
Pharmacy,
Charles
University,
Heyrovského
1203,
500
05
Hradec
Králové,
Czech
Republic,
2
Department
of
Microbi-‐
ology,
Nutrition
and
Dietetics,
Faculty
of
Agrobiology,
Food
and
Natural
Resources,
Czech
University
of
Life
Sciences,
Kamýcká
129,
165
21
Prague
6,
Czech
Republic,
3
Department
of
Biological
and
Biochemical
Sciences,
Faculty
of
Chemical
Technology,
University
of
Pardubice,
Studentská
573,
532
10
Pardubice,
Czech
Republic
Library
of
fourty-‐six
isoquinoline
alkaloids
possessing
eleven
structural
types
have
been
iso-‐
lated
in
our
laboratory,
and
their
cytotoxicity
against
human
epithelial-‐like
colorectal
adeno-‐
carcinoma
cells
Caco-‐2
and
hepatocellular
carcinoma
cells
Hep-‐G2
was
evaluated.
At
the
same
time,
normal
human
lung
fibroblast
cells
MRC-‐5
were
used
to
determine
overall
toxicity.
In
this
study
scoulerine
exhibited
potent
cytotoxic
potential
against
both
tested
cancer
cell
lines,
with
IC50
values
of
6.44
±
0.87
µM,
against
Caco-‐2
cells
and
4.57
±
0.42
µM
for
the
Hep-‐G2
cell
line.
Additionally,
berbamine
and
parfumidine
showed
cytotoxic
potential
against
the
Hep-‐G2
cell
line,
and
aromoline
showed
moderate
activity
against
the
Caco-‐2
cell
line;
the
other
eval-‐
uated
alkaloids
were
determined
as
non-‐cytotoxic
(IC50
>
100
μM)
against
the
tested
cell
lines.
Unfortunately,
scoulerine,
aromoline,
and
berbamine
also
showed
cytotoxicity
to
the
normal
cell
line
MRC-‐5.
Among
the
tested
isoquinoline
alkaloids,
scoulerine
was
selected
as
a
com-‐
pound
with
the
most
evident
cytotoxic
effects
in
cancer
cells
for
furthers
evaluations
to
eluci-‐
date
mechanisms
underlying
this
activity
and
for
SAR
study.
First
step
of
the
SAR
study
is
the
preparation
of
semisynthetic
analogues
by
changing
different
parts
of
the
structure
of
the
se-‐
lected
compound.
The
first
series
of
semisynthetic
analogues,
various
types
of
2,
9-‐di-‐O-‐esters
of
scoulerine
has
been
prepared.
All
ten
synthetic
analogues
were
evaluated
for
their
cytotox-‐
icity
against
Caco-‐2
cell
line.
Prepared
analogue
2,
9-‐di-‐O-‐hexanoylscoulerine
showed
approx-‐
imately
four
times
better
activity
against
Caco-‐2
cell
line
(IC50
=
1.75
±
0.41
µM)
compared
to
scoulerine.
All
compounds
are
assayed
for
their
toxicity
against
Hep-‐G2
cells,
and
also
overall
toxicity
against
noncancerous
cell
line
is
studied.
Acknowledgements:
This
project
was
supported
by
grants
SVV
UK
260
292,
Charles
University
grant
Nr.
17/2012/UNCE
and
Grant
Agency
of
the
Czech
University
of
Life
Sciences
Prague
CIGA
20132035.
Keywords: Isoquinoline alkaloids, scoulerine, semisynthetic analogues, cytotoxicity, Caco-‐2,
Hep-‐G2
P714
Isolation
and
structure
elucidation
of
twelve
new
prenylated
fla-‐
vonoids
from
Onobrychis
spp.
(Leguminosae)
employing
LC-‐
HRMS,
HSCCC
and
NMR
techniques
Maria-‐Eleni Sakavitsi, Job Tchoumtchoua, Sofia Mitakou, Maria Halabalaki, Alexios-‐Leandros
Skaltsounis
Division
of
Pharmacognosy
and
Natural
Products
Chemistry,
School
of
Pharmacy,
University
of
Athens,
15771,
Athens,
Greece
Onobrychis
spp.
(Leguminosae),
mostly
known
as
tanniferous
forage
legumes,
bear
great
eco-‐
nomic
impact
on
agricultural
and
livestock
fields.
In
our
previous
reports,
the
phytochemical
and
pharmacological
evaluation
of
Onobrychis
ebenoides,
a
Greek
endemic
plant,
yielded
novel
prenylated
compounds
with
remarkable
cytotoxic
and
estrogenic
activities
[1].
The
continua-‐
tion
of
our
studies
suggested
the
prenyl
moiety
position,
as
the
decisive
component
of
the
cy-‐
totoxicity
and/or
estrogenicity
in
a
cell-‐dependent
manner
[2,
3].
Therefore,
the
present
study
was
aimed
firstly
at
a
comparative
phytochemical
investigation
between
Onobrychis
ebenoides
and
Onobrychis
alba
subsp.
laconica,
another
endemic
species
of
Greece
without
scientific
re-‐
ports.
Among
these
lines,
LC-‐HRMS-‐based
dereplication
approaches
were
employed
for
tar-‐
geted
isolation
purposes.
Secondly,
both
extracts
subjected
to
isolation
and
purification
of
putative
new
compounds
using
High
Speed
Countercurrent
Chromatography
(HSCCC)
tech-‐
nique.
The
profiling
of
methanol
extracts
from
both
plants
was
performed
on
a
UHPLC-‐ESI
(-‐)-‐
LTQ-‐Orbitrap
system.
Chromatographic
and
spectrometric
features
were
incorporated
for
the
identification
procedure
[4].
Both
extracts
revealed
a
rich
content
in
prenylated
derivatives
of
isoflavonoids,
coumestans
and
benzofurans.
Consequent
targeted
fractionation
with
step-‐
gradient
HSCCC
afforded
5
new
isoflavonoids
namely
albalacone
I-‐II,
ebenosin
I-‐III,
4
new
coumestans
albalacol,
onobenol
I-‐III,
1
new
pterocarpan
onopterol
and
2
new
benzofurans
ebenfuran
IV
and
bis-‐ebenfuran
II,
as
indicated
in
the
figure
below,
along
with
10
known
compounds
belonging
to
the
respective
chemical
classes.
Conclusively,
both
Onobrychis
ebe-‐
noides
and
Onobrychis
alba
subsp.
laconica,
found
to
be
significantly
rich
in
at
least
three
dif-‐
ferent
classes
of
phytoestrogens,
known
for
their
estrogenic
as
well
as
their
cytotoxic
proper-‐
ties.
Keywords:
Onobrychis
ebenoides,
Onobrychis
alba
subsp.
laconica,
prenylated
flavonoids,
es-‐
trogenicity,
cytotoxicity
References:
[1] Papoutsi
Z,
Kassi
E,
Halabalaki
M,
Mitakou
S,
Moutsatsou
P.
Evaluation
of
estrogen-‐
ic/antiestrogenic
activity
of
Onobrychis
ebenoides
extract
-‐
Interaction
with
estrogen
re-‐
ceptor
subtypes
ERα
and
ERβ.
Toxicology
in
Vitro
2007;
3:
364-‐370
[2] Halabalaki
M,
Alexi
X,
Aligiannis
N,
Lambrinidis
G,
Pritsinis
H,
Florentin
I,
Mitakou
S,
Mikros
E,
Skaltsounis
A-‐L,
Alexis
MN.
Estrogenic
activity
of
isoflavonoids
from
On-‐
obrychis
ebenoides.
Planta
Med
2006;
6:
488-‐493
[3] Halabalaki
M,
Alexi
X,
Aligiannis
N,
Alexis
MN,
Skaltsounis
A-‐L.
Ebenfurans
IV-‐VIII
from
Onobrychis
ebenoides:
Evidence
that
C-‐prenylation
is
the
key
determinant
of
the
cytotoxi-‐
city
of
3-‐formyl-‐2-‐arylbenzofurans.
J
Nat
Prod
2008;
11:
1934-‐1937
[4] Tchoumtchoua
J,
Njamen
D,
Mbanya
JC,
Skaltsounis
AL,
Halabalaki
M.
Structure-‐oriented
UHPLC-‐LTQ
Orbitrap-‐based
approach
as
a
dereplication
strategy
for
the
identification
of
isoflavonoids
from
Amphimas
pterocarpoides
crude
extract.
J
Mass
Spectrom
2013;
48:
561-‐575
P715
B-‐ring
modification
in
prenylflavonoids
of
hops
and
the
effect
on
induction
of
neuronal
differentiation
Michael Kirchinger1, Corinna Urmann1, Lara Bieler2, Sebastien Couillard-‐Despres2, Herbert
Riepl1
1 Organic and Analytical Chemistry, Weihenstephan-‐Triesdorf University of Applied Sciences, Schulgasse
16
94315
Straubing,
Germany,
2
Institute
of
Experimental
Neuroregeneration
Paracelsus
Medical
Univer-‐
sity,
Salzburg,
Strubergasse
21,
5020
Salzburg,
Austria
Humulus
lupulus
L.
is
mostly
associated
to
brewing
industry
but
is
also
well
known
as
medical
plant.
Recently
the
prenylflavonoids
of
hop
were
identified
as
potent
inducers
of
neuronal
differentiation
in
adult
neuronal
stem
cells
[1,
2].
Therefore,
this
special
class
of
substances
is
discussed
as
possible
lead
structures
against
neurodegenerative
diseases.
Figure
3.
Xanthohumol
C,
A-‐and
B-‐ring.
Bioactivities
of
flavonoids
are
strongly
related
to
the
substitution
pattern
of
the
B-‐ring.
Flavo-‐
noids
with
two
hydroxyl
groups
in
ortho
position
for
instance
show
an
increased
anti-‐
oxidative
effect.
Accordingly,
we
investigated
the
influence
of
different
substitution
pattern
on
B-‐ring
of
this
special
class
of
flavonoids
concerning
the
induction
of
neuronal
differentiation.
Different
types
of
flavonoids
with
several
hydroxyl
and
ether
groups
were
synthesized
using
aldol
condensation.
In
the
next
step
the
oxygen
was
replaced
with
nitrogen
or
sulfur
leading
to
a
different
chemical
surrounding,
due
to
a
different
size
and
the
ability
for
hydrogen
bond
formation.
Subsequently,
the
activities
of
synthesized
compounds
were
quantified
using
a
du-‐
al
luciferase
reporter
gene
assay
based
on
doublecortin,
a
marker
of
early
neurons.
In
contrast
to
the
anti-‐oxidative
activity
a
second
hydroxyl
group
in
ortho
position
decreases
the
differentiation
inducing
activity.
In
addition,
blocking
of
hydroxyl
group
by
methylation
also
results
in
a
significant
decrease
of
activity.
Keywords:
Hop,
xanthohumol
c,
neuronal
differentiation
References:
[1]
Oberbauer
E,
Urmann
C,
Steffenhagen
C,
Bieler
L,
Furtner
T,
Humpel
C,
Baumer
B,
Bandtlow
C,
Couillard-‐Despres
S,
Rivera
FJ,
Riepl
H,
Aigner
L.
Chroman-‐like
cyclic
prenylflavonoids
promote
neuronal
differentiation
and
neurite
outgrowth
and
are
neuroprotective.
J
Nutr
Biochem
2013;
24:
1953-‐1962
[2]
Urmann
C,
Oberbauer
E,
Couillard-‐Despres
S,
Aigner
L,
Riepl
H.
Neurodifferentiating
poten-‐
tial
of
8-‐prenylnaringenin
and
related
compounds
in
neural
precursor
cells
and
correla-‐
tion
with
estrogen-‐like
activity.
Planta
Med
2015;
4:
305-‐311
P716
A
peptide
isolated
from
an
ant
active
against
Helicobacter
pylori
J.
Guzman1,
M.
Treilhou2,
D.
Castillo1,
H.
Belkhelfa3,
L.
Haddioui-‐Hbabi3,
M.
Sauvain1,4
1
Universidad
Peruana
Cayetano
Heredia
(UPCH),
Lima,
Peru,
2
Equipe
VacBio
EA
4357,
Université
Fédérale
Toulouse
Midi-‐Pyrénées,
CUFR
JF
Champollion,
Albi,
France,
3
Fonderephar,
Université
Fédérale
Toulouse
Midi-‐Pyrénées,
UPS,
Faculté
des
Sciences
Pharmaceutiques,
Toulouse,
France,
4
Université
Fédé-‐
rale
Toulouse
Midi-‐Pyrénées,
UPS,
PHARMADEV-‐UMR
152
IRD-‐UPS,
Toulouse,
France
An
antimicrobial
peptide,
named
Bicarinalin
[1],
isolated
from
the
venom
of
the
ant
Tetramo-‐
rium
bicarinatum
showed
remarkably
strong
activity
against
an
ATCC
strain
and
forty-‐four
Peruvian
patient
strains
of
Helicobacter
pylori
isolated
from
stomach
ulcer
biopsies.
Bicari-‐
nalin
had
a
potent
antibacterial
activity
(IC50
=
0.98
µM
at
the
same
magnitude
as
for
four
an-‐
tibiotics:
amoxicillin
IC50
<
0.07
µM,
clarithromycin
IC50
=
1.34
µM,
metronidazole
IC50
=
46.7
µM
and
levofloxacin
IC50
=
2.77
µM
used
currently
in
therapeutic
against
H.
pylori.
Moreover,
it
is
efficient
in
the
inhibition
of
H.
pylori
adherence
on
gastric
cells
with
an
IC50
=
0.098
µM
and
not
toxic
against
the
same
cells
(Selective
Index
>
7),
suggesting
potential
for
develop-‐
ment
into
an
anti-‐infective
agent
for
use
against
the
causal
agent
of
stomach
ulcers.
Bicari-‐
nalin
is
a
promising
lead
compound
in
the
search
for
more
effective
and
specific
H.
pylori
therapeutics.
Keywords: Antimicrobial peptide, Tetramorium bicarinatum, Bicarinalin, Helicobacter pylori,
stomach ulcers
References:
[1] Rifflet
A,
Gavalda
S,
Téné
N,
Orivel
J,
Leprince
J,
Guilhaudis
L,
Génin
E,
Vétillard
A,
Treilhou
M.
Identification
and
characterization
of
a
novel
antimicrobial
peptide
from
the
venom
of
the
ant
Tetramorium
bicarinatum.
Peptides
2012;
38:
363-‐370
P717
Anatomical
characterization
and
chemical
profiling
of
Rumex
species
Hye-‐Jin
Kim1,
Woo
Sung
Park1,
Ji-‐Yeong
Bae2,
Jong
Hee
Park3,
Mi-‐Jeong
Ahn1
1
College
of
Pharmacy
and
Research
Institute
of
Pharmaceutical
Sciences,
Gyeongsang
National
Universi-‐
ty,
Jinju
52828,
Korea,
2
National
Center
for
Natural
Products
Research,
University
of
Mississippi,
MS
38677,
USA,
3
College
of
Pharmacy,
Pusan
National
University,
Busan
46241,
Korea
Rumex
species
are
perennial
herbs
belonging
to
the
Polygonaceae
family,
which
are
widely
distributed
across
Eastern
Asia.
These
herbs
have
been
used
in
folk
medicine
to
treat
gastric
disease;
however,
the
botanical
origin
of
the
herbs
remains
to
be
completely
described
[1,
2].
In
this
study,
anatomical
characterization
and
chemical
profiling
were
carried
out
to
differen-‐
tiate
among
five
Rumex
species,
i.e.,
R.
acetosa,
R.
acetosella,
R.
crispus,
R.
japonicas,
and
R.
lon-‐
gifolius,
from
Korea.
The
inner
morphological
characteristics
of
these
specimens
were
ob-‐
served
using
transverse
or
vertical
sections,
which
were
prepared
using
a
microslicer
or
a
handslicer.
HPLC-‐DAD
analysis
was
preformed
to
verify
the
differences
in
chemical
profiles
among
the
species.
Druse
was
found
in
the
upper
surface
of
R.
acetosa
leaves
only.
R.
acetosa
and
R.
acetosella
showed
V-‐like
petioles,
while
the
other
species
displayed
elliptical
ones.
Fewer
vascular
bundles
were
observed
in
the
transverse
sections
of
the
petioles
of
R.
acetosa
and
R.
acetosella.
R.
longifolius
showed
the
highest
number
of
vascular
bundles
in
the
flower
stalk.
In
addition,
the
five
species
could
be
differentiated
by
other
internal
morphological
cri-‐
teria
such
as
the
shape
of
the
main
vein
of
leaves,
number
of
collenchyma
cell
layers,
and
the
frequency
of
stomata.
While
the
root
of
R.
crispus
showed
the
highest
content
(11.8
mg/g
DW)
of
the
three
major
anthraquinones
[3],
emodin,
chrysophanol
and
physcion,
it
also
exhibited
the
lowest
content
(1.92
mg/g
DW)
of
three
anthraquinone
glycosides.
In
contrast,
while
the
content
(1.12
mg/g
DW)
of
the
above
three
anthraquinones
was
lowest
in
the
root
of
R.
ace-‐
tosa,
this
species
exhibited
a
high
content
(13.6
mg/g
DW)
of
the
glycosides.
R.
acetosella
par-‐
ticularly
exhibited
a
high
content
of
the
anthraquinones
(6.94
mg/g
DW)
and
their
anthraqui-‐
none
glycosides
(21.8
mg/g
DW).
These
significantly
different
aspects,
including
other
pa-‐
rameters
in
HPLC
profiles,
should
be
key
factors
to
differentiate
among
the
five
Rumex
spe-‐
cies.
Acknowledgements:
This
research
was
supported
by
the
Basic
Science
Research
Program
through
the
National
Research
Foundation
of
Korea
(NRF)
funded
by
the
Ministry
of
Science,
ICT
&
Future
Planning
(NRF-‐2014R1A1A305070)
and
the
Ministry
of
Trade,
Industry
&
Energy
(MOTIE),
Korea
Institute
for
Advancement
of
Technology
(KIAT)
through
the
Inter-‐ER
Cooperation
Projects
(Grant
#
A004500005).
Keywords: Rumex species, anatomical characteristics, HPLC profiles, anthraquinones, an-‐
thraquinone glycosides
References:
[1] Bae
J-‐Y,
Lee
YS,
Han
SY,
Jeong
EJ,
Lee
MK,
Kong
JY,
Lee
DH,
Cho
KJ,
Lee
H-‐S,
Ahn
M-‐J.
A
com-‐
parison
between
water
and
ethanol
extracts
of
Rumex
acetosa
for
protective
effects
on
gastric
ulcers
in
mice.
Biomol
Ther
2012;
20:
425-‐430
[2] Lee
NJ,
Choi
JH,
Koo
BS,
Ryu
SY,
Han
YH,
Lee
SI,
Lee
DU.
Antimutagenicity
and
cytotoxicity
of
the
constituents
from
the
aerial
parts
of
Rumex
acetosa.
Biol
Pharm
Bull
2005;
28:
2158-‐2161
[3] Lim
JP,
Park
Y-‐S,
Hong
MW,
Kim
DK.
Quantitative
analysis
of
anthraquinones
from
the
roots
of
Korean
natural
Rumex
species
plants.
Kor
J
Pharmacogn
2011;
42:
297-‐301
P718
Volatile
compounds
of
Tripleurospermum
decipiens
from
differ-‐
ent
sites
in
Turkey
Mine
Kurkcuoglu1,
Fatma
Tosun2,
Huseyin
Inceer3,
K.
Husnu
Can
Baser1,4
1
Anadolu
University,
Faculty
of
Pharmacy,
Department
of
Pharmacognosy,
TR-‐26470
Eskisehir,
Turkey,
2
Gazi
University,
Faculty
of
Pharmacy,
Department
of
Pharmacognosy
TR-‐06330Ankara,
Turkey,
3
Ka-‐
radeniz
Technical
University,
Faculty
of
Sciences
and
Arts,
Department
of
Biology,
TR-‐61080
Trabzon,
Turkey,
4
Near
East
University,
Faculty
of
Pharmacy,
Department
of
Pharmacognosy,
Near
East
Boule-‐
vard,
99138,
Nicosia
/
TRNC,
Mersin
10,
Turkey
The
genus
Tripleurospermum
Schultz
Bip.
is
represented
in
Turkey
by
31
taxa,
15
being
en-‐
demic
in
Turkey[1-‐3].
Hydrodistilled
essential
oils
of
the
dried
flowers
of
Tripleurospermum
decipiens
(Fisch.
&
Mey.)
Bornm.
[Asteraceae]
were
analyzed
by
GC-‐FID
and
GC-‐MS.
The
oils
were
characterized
by
the
occurrence
of
matricaria
esters
as
main
constituents.
Flowers
of
T.
decipiens
were
collected
from
two
localities
in
Turkey;
A:
Adana:
Saimbeyli-‐Yesilkent
road,
12.06.2014–TDA;
B:
Eskişehir:
Between
Bozüyük-‐
Eskişehir,
26.06.2009–TDB.
The
essential
oils
of
TDA
and
TDB
were
characterized
by
the
occurrence
of
(2Z,8Z)-‐matricaria
ester
(TDA:
57.9%;TDB:
70.0%),
β-‐sesquiphellandrene
(TDA:
10.4%;
TDB:
2.7%),
(2E,8Z)-‐matricaria
ester
(TDA:
8.1%;TDB:
1.7%),
(Z)-‐β-‐farnesene
(TDA:
7.5%,
TDB:
0.3%),
(2E,8E)-‐matricaria
ester
(TDA:
2.3%,
TDB:
0.4%)
and
(2Z,8E)-‐matricaria
ester
(TDA:
1.4%;
TDB:
0.5%)
as
main
con-‐
stituents.
Matricaria
esters
were
previously
found
in
other
Tripleurospermum
species
such
as
T.
disciforme
(C.A.Mey)
Schultz-‐Bip.
[5]
and
Matricaria
perforata
Merat
(=Tripleurospermum
inodorum
(L.)
Sch.
Bip)
[6].
To
the
best
of
our
knowledge,
this
is
the
first
report
on
the
essen-‐
tial
oil
composition
of
Tripleurospermum
decipiens.
Keywords:
Tripleurospermum
decipiens,
essential
oils,
GC,
GC-‐MS,
matricaria
esters
References:
[1] Davis
PH.
Flora
of
Turkey
and
the
East
Aegean
Islands.
Edinburgh:
Edinburgh
University
Press,
1975;
5:
295-‐311
[2] Inceer
H,
Hayirlioglu-‐Ayaz
S.
Tripleurospermum
ziganaense
(Asteraceae,
Anthemideae),
a
new
species
from
north-‐east
Anatolia,
Turkey.
Bot
J
Linn
Soc
2008;
158:
696-‐700
[3] Inceer
H,
Hayirlioglu-‐Ayaz
S.
Tripleurospermum
insularum
(Asteraceae,
Anthemideae),
a
new
species
from
Turkey.
Ann
Bot
Fennici
2014;
51:
49-‐53
[4] Jaimand,
K,
Rezaee,
M.B.
Investigation
extraction
by
two
different
apparatus
and
effects
of
essential
oils
on
content
and
constituents
of
Tripleurospermum
disciforme(C.A.Mey)
Schultz-‐Bip.
Pajouhesh-‐va-‐Sazandegi.
In
Natural
Sciences,
2003;
60:
2-‐7
[5] NazarAlipour,
A,
,Sefidkon,
F.
Quantitative
and
qualitative
study
of
the
essential
oil
from
aromatic
and
medicinal
Tripleurospermum
disciforme
(C.A.
Mey.)
Schultz
-‐
Bip.
J
Med
Plants
2003;
2:
33-‐40
[6] Raal
A,
Kaur
H,
Orav
A,
Arak
E,
Kailas
T,
Müürisepp
M.
Content
and
Composition
of
Essen-‐
tial
Oils
in
Some
Asteraceae
Species,
Proceedings
of
the
Estonian
Academy
of
Sciences
2011;
60:
55-‐63
P719
Phytochemical
and
cytotoxic
studies
on
Cestrum
nocturnum
Mona
A.
Mohamed
Department
of
Medicinal
Chemistry,
Theodor
Bilharz
Research
Institute,
Kornish
El-‐Nile,
1266,
Warrak
El-‐Hadar,
Giza,
Egypt
The
genus
Cestrum
belonging
to
family
solanaceae.
Several
Cestrum
species
have
already
been
chemically
studied
and
found
to
contain
mainly
steroidal
saponin
[1],
triterpenes
[2]
and
fla-‐
vonoids
[3].
From
the
leaves
of
Cestrum
nocturnum
(solanaceae)
two
new
triterpene
saponins,
named
sophradiol
3-‐
O-‐
α
-‐L-‐1C4-‐rhamnopyranosyl-‐(1'''→4'')-‐
O-‐β
-‐D-‐4C1-‐glucopyranosyl
(1''→6')-‐
O-‐β-‐D
4C1-‐
galactopyranoside
(10)
and
23-‐hydroxy-‐3α-‐[(O-‐α-‐L-‐1C4-‐
rhamnopyranosyl-‐(1"→2')-‐O-‐α-‐L4C1-‐arabinopyranosyl)
oxy]olean12-‐en-‐28-‐oic
acid
O-‐α-‐L-‐1C4-‐
rhamnopyranosyl-‐(1'''''→4'''')-‐O-‐β-‐D-‐4C1-‐glucopyranosyl-‐(1''''→6''')-‐O-‐
D-‐4C1-‐
galactopyranosyl
ester
(11)
were
isolated.
In
addition,
nine
known
compounds
were
isolated,
namely
E-‐
caffeic
acid
(1),
quercetin
(2),
4',5-‐dimethoxy
quercetin
(3),
8-‐
sulfated
apigenin
(4)
rutin
(5),
quercetin
3-‐O-‐β-‐D-‐4C1-‐neohesperidoside
(6),
3,3',4'-‐
trimethoxy
quercetin
(7),
oleanolic
acid,
(8),
oleanolic
acid
3-‐O-‐
β-‐D-‐4C1-‐glucopyranosyl(1''→3')-‐
O-‐β-‐D-‐4C1-‐
glucopy-‐
ranside
(9),
All
metabolites
were
isolated
for
the
first
time
from
this
plant.
The
structures
were
determined
mainly
by
spectroscopic
methods
(UV,
ESI-‐MS,
1H-‐,
13C-‐NMR,1H-‐1H
COSY,
HSQC
and
HMBC).
Cytotoxic
screening
of
the
butanol,
ethyl
acetate
and
chloroform
extracts
was
carried
out
on
brine
shrimps.
In
addition
the
investigated
butanol
extract
and
major
iso-‐
lates
(10
and
11)
were
also
tested
against
the
HepG2
tumor
cell
line.
Keywords: Cestrum nocturnum, solanaceae, brine shrimp, cytotoxic, HepG2
References:
[1] Ahmad VU, Baqai FT, Ahmad R. Diosgenin Tetrasaccharide from Cestrum nocturnum. Z.
Naturforsch 1995; 50: 1104-‐1110
[2] Hamed M, El-‐Amin S, Abdel-‐Fattah AS. Molluscicidal activity of some constituentsisolated
from Cestrum purpureum. Pharmacologyonline 2015; 2: 59-‐72
[3] D'Abrosca B , DellaGreca M , Fiorentino A , Monaco P , Zarrelli A. Low molecular weight
phenols from the bioactive aqueous fraction of Cestrum parqui. J Agric Food Chem 2004;
52: 4101-‐4108
P720
Bioactive
formylated
flavonoids
from
Eugenia
rigida:
Their
isola-‐
tion,
synthesis,
and
X-‐ray
crystallography
Muhammad
Ilias1,
Mohamed
A.
Zaki2,
Melissa
R.
Jacob1,
Shabana
I.
Khan1,3,
Mohamed
A.
Ibra-‐
him1,
Volodymyr
Samoylenko1,
Rabab
Mohammed2,
Mona
H.
Hetta2,
David
D.
Pasco1,
Daneel
Ferreira1,3,
Frank
R.
Fronczek4,
Dhammika
Nanayakkara1
1
National
Center
for
Natural
Products
Research,
Research
Institute
of
Pharmaceutical
Sciences,
School
of
Pharmacy,
University
of
Mississippi,
University,
MS
38677,
USA,
2
Department
of
Pharmacognosy,
School
of
Pharmacy,
Beni-‐Suef
University,
Beni-‐Suef,
Egypt,
3
Department
of
Biomolecular
Sciences,
School
of
Pharmacy,
University
of
Mississippi,
University,
MS
38677,
USA,
4
Department
of
Chemistry,
Louisiana
State
University,
Baton
Rouge,
LA
70803-‐1804,
USA
Two
new
formylated
flavonoids,
1
and
2,
together
with
their
known
derivatives
5,7-‐
dihydroxy-‐8-‐formylflavanone
(3),
4΄,6΄-‐dihydroxy-‐2΄-‐methoxy-‐3΄-‐methyldihydrochalcone
(4),
7-‐hydroxy-‐5-‐methoxy-‐6-‐methylflavanone
(5),
3΄-‐formyl-‐2΄,4΄,6΄-‐trihydroxy-‐5΄-‐methyl-‐
dihy-‐
drochalcone
(6)
and
3΄-‐formyl-‐2΄,4΄,6΄-‐trihydroxydihydrochalcone
(7)
were
isolated
using
centrifugal
preparative
TLC
[1]
from
the
leaves
of
Eugenia
rigida
DC.
(Fam.
Myrtaceae).
The
individual
(S)
and
(R)-‐enantiomers
of
1
and
3,
together
with
the
corresponding
formylated
flavones,
12
(6-‐formyl-‐5,7-‐dihydroxyflavone)
and
13
(8-‐formyl-‐5,7-‐dihydroxyflavone),
as
well
as
3΄-‐formyl-‐2΄,4΄,6΄-‐trihydroxychalcone
(15)
and
the
dihydrochalcone
7
were
synthe-‐
sized.
The
structures
of
the
isolated
and
synthetic
compounds
were
established
by
full
spec-‐
troscopic
data,
including 2D
NMR
experiments
and
HRMS.
In
addition,
structures
3,
5,
and
12
were
confirmed
by
single
crystal
X-‐ray
crystallography.
The
isolated
and
synthetic
flavonoids
were
evaluated
for
antimicrobial,
antifungal,
and
cytotoxic
activities
against
selected
microor-‐
ganisms
and
solid
tumor
cells,
respectively.
Antibacterial
activities
against
Staphylococcus
aureus
and
methicillin-‐resistant
S.
aureus
were
observed
for
compound
7
(IC50/MIC
of
4.99/20
µg/mL
and
10.49/20
µg/mL,
respectively),
and
antifungal
activity
for
15
against
Candida
albicans,
C.
glabrata,
C.
krusei
and
Cryptococcus
neoformans
(IC50/MIC
of
18.58/20,
2.86/20
and
9.72/20,
and
6.98/20
µg/mL),
while
7
was
active
against
C.
glabrata
and
Cr.
neoformans
(IC50
3.25
and
3.03
µg/ml,
respectively).
Acknowledgements:
This
work
was
supported
by
NCNPR,
and
in
part
by
the
United
States
Department
of
Agriculture
ARS
cooperative
agreement
No.
58-‐6408-‐2-‐0009.
MAZ
is
thankful
to
the
Egyptian
Govt.
for
scholarship.
Keywords: Eugenia rigida, flavonoids, synthesis
References:
[1] Ilias
M,
Samoylenko
V,
Gillum
VD.
Preparation
of
pre-‐coated
rp-‐rotors
and
universal
chromatorotors,
chromatographic
separation
devices
and
methods
for
centrifugal
pre-‐
parative
chromatography.
USPTO
Application
filed
March
7,
2014,
Serial
No.
USSN
14/343,830
(Patent
Pending)
P721
Flavone
derivatives:
a
promising
tool
in
the
fight
against
malaria
Flore
Nardella1,2,
Silvia
Stiebing3,
Patrick
Wagner1,
Valérie
Collot3,
Marcel
Kaiser4,
Benoit
Witkowski5,
Didier
Menard5,
Martine
Schmitt1,
Ermanno
Candolfi2,
Catherine
Vonthron-‐
Sénécheau1
1
Equipe
Chimie
Biologie
Intégrative
-‐
Laboratoire
d’Innovation
Thérapeutique,
UMR
CNRS-‐Unistra
7200,
Faculté
de
Pharmacie,
74
route
du
Rhin,
CS60024,
67401
Illkirch
Cedex,
France,
2
Institut
de
Parasitologie
et
de
Pathologie
Tropicale
de
Strasbourg,
Faculté
de
Médecine,
3
rue
Koeberlé,
67000
Strasbourg,
France,
3
Centre
d’Etudes
et
de
Recherches
sur
le
Médicament
en
Normandie,
Université
de
Caen
Basse-‐
Normandie,
Boulevard
Becquerel,
14032
Caen
Cedex,
France,
4
Swiss
Tropical
and
Public
Health
Institute,
Socinstrasse
57,
4051
Basel,
Switzerland,
5
Unité
d’Epidémiologie
Moléculaire
du
Paludisme,
Institut
Pas-‐
teur
du
Cambodge,
5
Boulevard
Monivong,
Phnom
Penh,
Cambodia.
P.
falciparum
malaria
is
the
deadliest
parasitic
disease
with
438.000
deaths
every
year
[1].
The
increasing
resistance
of
Plasmodium
to
antimalarials,
notably
to
the
first
line
treatment,
artemisinins
[2],
is
a
major
threat
and
fast
acting
drugs
with
new
mechanisms
of
action
are
needed.
We
isolated
an
active
biflavonoid
from
Campnosperma
panamense
(Anacardiaceae,
IC50
=
480
nM
in
vitro
on
P.
falciparum
K1
multi-‐resistant
strain)
[3],
and
developed
novel
simplified
synthetic
analogs
(MR
series)
with
improved
pharmacological
and
pharmacokinet-‐
ic
profiles.
One
of
these
analogs,
MR70
is
parasiticidal
on
early
blood
stages
of
P.
falciparum
in
less
than
6
hours.
Moreover,
MR70
and
its
analog
MR87
exhibit
a
partial
in
vivo
antimalarial
activity,
reducing
parasitemia
by
35%
to
70%,
respectively,
on
day
4
in
a
murine
model
(P.
berghei
ANKA,
100
mg/kg
for
4
days).
But
these
compounds
showed
no
significant
improve-‐
ment
in
terms
of
survival.
A
structure-‐activity
relationship
study
is
still
ongoing
to
further
improve
these
results.
MR70
is
a
fast
acting
drug.
To
our
knowledge,
it
is
the
only
compound
targeting
specifically
these
stages,
suggesting
a
potential
new
mechanism
of
action.
Interest-‐
ingly,
this
stage
is
specifically
the
one
that
is
resistant
to
artemisinins
[4]
and
we
are
currently
assessing
MR70
efficacy
on
resistant
isolates.
Further
investigation
is
needed
to
optimize
MR70
activity
and
to
understand
its
underlying
mechanism(s)
of
action.
Acknowledgements:
Fondation
Groupe
Pasteur
Mutualité,
Fondation
Pierre
Ledoux
Jeunesse
Internatio-‐
nale,
Réseau
International
des
Instituts
Pasteur.
Keywords: Plasmodium, flavone, in vivo, artemisinin, resistance, fast acting drug
References:
[1] World malaria report, 2015, WHO. http://www.who.int/malaria/publications/world-‐
malaria-‐report-‐2015/report/en/
[2] Noedl H, Se Y, Schaecher K, Smith BL, Socheat D, Fukuda MM. Evidence of artemisinin-‐
resistant malaria in western Cambodia. N Engl J Med 2008; 24: 2619 – 2620
[3] Weniger B, Vonthron-‐Sénécheau C, Arango GJ, Kaiser M, Brun R, Anton R. A bioactive bi-‐
flavonoid from Campnosperma panamense. Fitoterapia 2004; 7: 764 – 767
[4] Witkowski B, Amaratunga C, Khim N, Sreng S, Chim P, Kim S, Lim P, Mao S, Sopha C, Sam
B, Anderson J, Duong S, Chuor CM, Taylor WRJ, Suon S, Mercereau-‐Puijalon O, Fairhurst
RM, Menard D. Novel phenotypic assays for the detection of artemisinin-‐resistant Plas-‐
modium falciparum malaria in Cambodia: in-‐vitro and ex-‐vivo drug-‐response studies.
Lancet Infect Dis 2013; 12: 1043 – 1049
P722
A two-‐season impact study on Globularia alypum: Adaptive leaf
structures and metabolic variations
Stavroula Mamoucha1, Nikolaos Tsafantakis2, Nikolas Fokialakis2, Nikolaos Christodoulakis1
1 Faculty of Biology, University of Athens, Panepistimioupolis ,15771, Athens, Greece, 2 Depart-‐
ment
of
Pharmacognosy
and
Natural
Products
Chemistry,
Faculty
of
Pharmacy,
University
of
Athens,
Panepistimioupolis
-‐
15771
Athens
,
Greece
Globularia
alypum
is
a
phryganic
species
in
a
genus
composed
of
various
herbs
or
shrubs
growing
in
Europe,
in
particular
in
the
Mediterranean
region.
It
is
well
known
that
plants
of
the
Mediterranean
climate
undergo
to
several
abiotic
and
biotic
stressful
conditions,
thus
Mediterranean
plant
species
have
evolved
adaptations
to
maximize
tolerance.
Among
these
adaptations,
anatomical
structures
and
biochemical
modifications
are
widely
reported
[1].
Secondary
metabolites
are
produced
by
plants
not
only
for
defense
purposes
against
pathogens,
but
also
as
protection
agents
against
environmental
stresses
[2].
In
the
present
study,
a
plant
anatomical
and
phytochemical
investigation
on
G.
alypum
was
carried
out,
in
order
to
examine
the
adaptive
structures
and
the
metabolic
variations
occurred
in
response
to
the
different
climate
conditions.
Summer
and
winter
leaves
were
detached,
fixed,
sectioned
and
investigated
using
light,
transmission
and
scanning
electron
microscopy
along
with
histo-‐
chemical
tests.
UHPLC-‐HRMS
analysis
was
carried
out
in
order
to
define
the
nature
and
the
relative
abundance
of
SM
under
the
two
different
seasons.
The
anatomical
adaptations
of
G.
alypum
evolve
a
compact,
isolateral,
amphistomatic
summer
leaf,
which
appears
rather
xero-‐
morphic,
and
epidermal
cells
which
posses
a
thick
external
wall,
accumulate
secondary
me-‐
tabolites
and
host
numerous
small,
capitate
hairs.
Mesophyll
cells
are
characterized
by
the
accumulation
of
osmiophillic
metabolites
within
their
vacuole.
In
the
phytochemical
study,
iridoids,
flavonoids
and
phenyl
ethanoid
glucosides
were
mainly
identified.
Among
the
differ-‐
ent
classes,
(E)-‐globularisin
showed
the
higher
amount
during
the
summer
season
while
ver-‐
bascoside
was
3-‐fold
higher
during
winter
[3,
4].
On
the
other
hand,
flavonoids
relative
con-‐
tent
was
less
remarkable,
among
seasons.
Interestingly,
winter
leaves
are
mostly
like
the
summer
ones
yet
a
bit
thicker
while
the
accumulation
of
secondary
metabolites
belonging
resulted
far
inferior.
Acknowledgements:
This
work
was
supported
by
IKY
-‐
State
Scholarship
Foundation,
Athens,
Greece.
Keywords: Globularia alypum, leaf anatomy, secondary metabolites, UHPLC-‐HRMS
References:
[1] Llusia
J,
Pen˜
uelas
J,
Alessio
GA
and
Estiarte
M.
Seasonal
contrasting
changes
of
foliar
con-‐
centrations
of
terpenes
and
other
volatile
organic
compound
in
four
dominant
species
of
a
Mediterranean
shrubland
submitted
to
a
field
experimental
drought
and
warming.
Physiol
Plant
2006;
127:
632-‐649
[2] Arbona
V,
Manzi
M,
Gómez-‐Cadenas
C,
Ollas
and
Gómez-‐Cadenas
A.
Metabolomics
as
a
tool
to
investigate
abiotic
stress
tolerance
in
plants.
Int
J
Mol
Sci
2013;
14:
4885-‐4911
[3] Es-‐Safi
N-‐E,
Kollmann
A,
Khlifi
S,
Ducrot
PH.
Antioxidative
effect
of
compounds
isolated
from
Globularia
alypum
L.
structure–activity
relationship.
LWT-‐Food
Sci
Technol
2007;
40:
1246-‐1252
[4] Amessis-‐Ouchemoukh
N,
Abu-‐Reidah
MI,
Quirantes-‐Piné
R,
Rodríguez-‐Pérez
C,
Madani
K,
Fernández-‐Gutiérrez
A,
Segura-‐Carretero
A.
Tentative
characterisation
of
iridoids,
phe-‐
nylethanoid
glycosides
and
flavonoid
derivatives
from
Globularia
alypum
L.
(Globularia-‐
ceae)
leaves
by
LC-‐ESI-‐QTOF-‐MS.
Phytochem
Anal
2014;
25:
389-‐484
P723
Synthesis
and
thymidine
phosphorylase
inhibition
studies
of
5-‐
chlorobenzothiazole
derivatives
Mastura
Arbin1,2,
Norizan
Ahmat1,2,Muhammad
Taha1,2
1
Faculty
of
Applied
Sciences,
Universiti
Teknologi
MARA
(UiTM),
Shah
Alam,
40450
Selangor
Darul
Ehsan,
2
Atta
ur
Rahman
Institute
for
Natural
Products
Discovery,
Universiti
Teknologi
MARA,
UiTM
Pun-‐
cak
Alam,
42300
Bandar
Puncak
Alam,
Selangor,
Malaysia
Benzothiazole
and
their
derivatives
have
been
previously
reported
to
exhibit
antitumor,
an-‐
timicrobial,
antiviral
and
anticonvulsant
activities
[1].
Thymidine
phosphorylase
inhibitors
have
attracted
great
attention
due
to
their
ability
to
suppress
the
tumors
formation.
In
our
ongoing
research,
a
series
of
5-‐chlorobenzothiazole
derivatives
(1–10)
have
been
synthesized
in
good
to
excellent
yields
(80–90%)
and
their
thymidine
phosphorylase
inhibition
potential
has
also
been
evaluated.
The
synthesized
compounds
showed
moderate
thymidine
phosphor-‐
ylase
inhibitory
activity
with
IC50
values
ranging
from
19.60
±
0.45
to
93.50±
2.88µM
,
and
7-‐
deazaxanthine
(7DX)
was
used
as
a
standard(IC50
38.68
±
4.42).
Compound
1,4-‐(5-‐
chlorobenzo[d]thiazol-‐2-‐yl)benzene-‐1,3-‐diol
and
compound
2,
5-‐chloro-‐2-‐(4-‐
chlorophenyl)benzo[d]thiazole
showed
the
lowest
IC50
value
of
19.60
±
0.45
µM
and
23.40
±
0.68
µM,
compared
to
the
standard
inhibitor.
These
compound
showed
better
phosphorylase
inhibition
activity
as
compared
to
7-‐deazaxanthine
[2].
HO
Cl N
Cl N
OH
S Cl
S
4-‐(5-‐chlorobenzo[d]thiazol-‐2-‐yl)benzene-‐1,3-‐diol
5-‐chloro-‐2-‐(4-‐chlorophenyl)benzo[d]thiazole
1
2
Figure
1.
Structure
compound
of
Benzothiazole
Acknowledgements:
Faculty
of
Applied
Sciences,
Universiti
Teknologi
MARA
(UiTM)
and
Atta
ur
Rahman
Institute
for
Natural
Products
Discovery,
are
acknowledged
for
providing
laboratory
facilities.
Keywords: 5-‐chlorobenzothiazole, pyiridine, heterocyclic, pharmacological
References:
[1] Srivastava
SK,
Yadav
R,
Srivastava
SD.
Synthesis
and
biological
activity
of
4-‐
oxothiazolidines
and
their
5-‐arylidenes.
Indian
J
Chem
2004;
43:
399-‐405
[2] Sohail
AS,
Shahzad
A,
Yar
M
,
Bajda
M,
Jadoon
B,
Khan
ZA,
Naqvi
SAR,
Shaikh
AJ
,
Hayat
K
,
Mahmmod
A,
Mahmood
N,
Filipek
S.
Synthesis
and
biological
evaluation
of
novel
oxadia-‐
zole
derivatives:
A
new
class
of
thymidine
phosphorylase
inhibitors
as
potential
anti-‐
tumor
agents.
Bioorg
Med
Chem
2014;
22:
1008-‐1015
P724
Lupeol
and
resveratrol
from
the
stembark
of
Shorea
ovalis
(Dip-‐
terocarpaceae)
Rosmawati
A.
Aziz1,2,
N.
Ahmat1,2
1
Faculty
of
applied
Sciences,
Universiti
Teknologi
MARA,
40450
Shah
Alam,
Selangor,
Malaysia,
2
Atta-‐ur-‐
Rahman
Institute
for
Natural
Product
Discovery,
Universiti
Teknologi
MARA,
Puncak
Alam
Campus,
42300
Bandar
Puncak
Alam,
Selangor
Darul
Ehsan,
Malaysia
Dipterocarpaceae
is
a
plant
family
of
large
three
that
is
widely
distributed
in
the
tropical
rain
forest
of
Southeast
Asia,
such
as
Borneo
and
Peninsular
Malaysia.
Shorea
is
the
largest
genus
of
Dipterocarpaceae
[1].
This
genus
is
the
most
economically
important
in
Malaysia.
The
tim-‐
bers
produced
are
very
diverged,
ranging
from
light
to
very
heavy,
and
are
characterized
by
the
presence
of
intercellular
resin
canals
with
color
ranging
from
white,
yellow,
pink,
dark-‐red
to
brown.
In
Malaysia,
Shorea
is
known
as
Balau,
Meranti
Pa’ang,
Meranti
Damar
Hitam
and
Red
Meranti.
Shorea
ovalis
is
the
species
included
in
the
group
of
Red
Meranti.
Locally
it
is
known
as
Meranti
Kepong.
So
far,
very
limited
chemical
constituents
have
been
reported
from
Shorea
ovalis.
In
this
study,
a
phytochemical
study
was
conducted
on
the
stem
bark
of
Shorea
ovalis.
The
stem
bark
was
obtained
from
Hutan
Simpan,
Jengka,
Pahang.
The
cleaned,
chopped
and
dried
stem-‐bark
was
grinded
into
powder.
The
powder
was
extracted
using
acetone
for
several
times.
The
separation
of
the
components
was
carried
out
using
vacuum
liquid
chro-‐
matography
(VLC)
followed
by
radial
chromatography
and
preparative
thin
layer
chromatog-‐
raphy
method.
The
structure
of
the
pure
compounds
was
elucidated
by
spectroscopic
meth-‐
ods
including
nuclear
magnetic
resonance
(NMR),
UV-‐
Vis,
and
comparison
with
literature.
Two
compounds
were
isolated
and
identified
as
lupeol
[2,
3]
and
resveratrol.
These
com-‐
pounds
were
reported
for
the
first
time
from
this
plant.
Acknowledgements:
We
thank
the
Faculty
of
Applied
Sciences,
Universiti
Teknologi
MARA,
Shah
Alam
for
providing
the
laboratory
facilities
to
do
the
research.
Keywords: Lupeol, resveratrol, Shorea ovalis, Dipterocaparceae, NMR
References:
[1] Noviany.
The
Isolation
of
α-‐viniferin,
A
Trimer
Stilbene,
from
Shorea
ovalis
Blume.
Mo-‐
dern
Applied
Science
2009;
3:
45-‐51
[2] Pattamadilok
D,
Suttisri
R.
Seco-‐terpenoid
and
other
constituents
from
Elateriospermum
tapos.
J
Nat
Prod
2008;
71:
292-‐294
[3] Jamal
Ak,
Yaacob
WA,
Din
LB.
A
chemical
Study
on
Phyllanthus
reticulatus.
Journal
of
Physical
Sciences
2008;
19:
45-‐50
P725
Synthesis
of
benzimidazole
derivatives
as
new
α-‐Glucosidase
in-‐
hibitors
Nik
Khairunissa
Nik
Abdullah
Zawawi1,2,
Norizan
Ahmat1,2,
Muhammad
Taha1,2,
Aisyah
Sali-‐
hah
Kamarozaman2,3
1
Atta-‐ur-‐Rahman
Institute
for
Natural
Product
Discovery,
Universiti
Teknologi
MARA
(UiTM),
Puncak
Alam
Campus,
42300
Bandar
Puncak
Alam,
Selangor
D.
E.
Malaysia,
2
Faculty
of
Applied
Science,
UiTM
Shah
Alam,
40450
Shah
Alam,
Selangor
D.E.
Malaysia,
3
Centre
of
Foundation
Studies,
Universiti
Teknolo-‐
gi
MARA,
Selangor
Branch,
Dengkil
Campus,
43800
Dengkil,
Selangor,
Malaysia
Synthesis of 4-‐(5,6-‐dimethyl-‐1H-‐benzo[d]imidazole-‐2-‐yl)benzohydrazide Schiff bases (1-‐26)
was accomplished by three-‐step procedure as shown in the scheme below. Equimolar solution
of 4-‐formylbenzoic acid methyl ester, 4,5-‐dimethyl-‐O-‐phenylenediamine and sodium meta
bisulfite in dimethylformamide (DMF) was refluxed for six hours to give out the arylester sub-‐
stituted benzimidazole [1]. The benzohydrazide of benzimidazole was formed by refluxing
arylester of benzimidazole with methanolic hydrazine hydrate. The synthesis of new benzim-‐
idazole benzohydrazide Schiff bases (1-‐26) was accomplished by reacting different aldehydes
with benzimidazole benzohydrazide in n-‐butanol in the presence of catalytic amount of acetic
acid and were further evaluated for their α-‐glucosidase inhibitory activity. Compounds 1-‐26
exhibited varying degrees of yeast α-‐glucosidase inhibitory activity with IC50 values between
8.40 ± 0.76 -‐ 179.71 ± 1.11 μM when compared with standard acarbose. In this assay, seven
compounds that showed highest inhibitory effects than the rest of benzimidazole series were
identified. All the synthesized compounds were characterized by different spectroscopic
methods adequately such as 1D NMR, IR and HREIMS.
NH2
O O NH2 Na S O ,
DMF, N O
2 2 5
+
O H reflux
6
hours N O
H
Hydrazine
RCHO
MeOH
4 3
2' 3' Acetic
acid,
n-‐butanol
N O reflux
3
hours N O
N HN N N HN NH2
7 H 6' 5' H
1 R
(1-‐26)
Acknowledgements:
The
authors
would
like
to
acknowledge
the
Ministry
of
Higher
Education
(MOHE)
and
Universiti
Teknologi
MARA
for
the
financial
support
under
RAGS
grant
600-‐RMI/RAGS/5/3/
(2/2012)
Keywords: Synthesis, benzimidazole, α-‐Glucosidase inhibitory activity
References:
[1] Taha
M,
Ismail
NH,
Jamil
W,
Rashwan
H,
Kashif
SM,
Sain
AA,
Adenan
MI,
Anouar
EH,
Ali
M,
Rahim
F,
Khan
KM.
Synthesis
of
novel
derivatives
of
4-‐methylbenzimidazole
and
evalua-‐
tion
of
their
biological
activities.
Eur
J
Med
Chem
2014;
84:
731-‐738
P726
New
cassane
diterpenes
from
the
leaves
and
twigs
of
Caesalpinia
bonduc
(Linn)
Roxb
Olubanke
O.
Ogunlana1,
Wen
J.
He2,
Jun
T.
Fan2,
Guang
Z.
Zeng2,
Oluseyi
E.
Ogunlana3,
Abiodun
H.
Adebayo1,
Chang
J.
Ji2,
Joseph
O.
Olagunju4,
Afolabi
A.
Akindahunsi5
and
Ning
H.
Tan2
1
Department
of
Biological
Sciences,
College
of
Science
and
Technology,
Covenant
University,
PMB
1023,
Ota,
Ogun
State,
Nigeria,
2
State
Key
Laboratory
of
Phytochemistry
and
Plant
Resources
in
West
China,
Kunming
Institute
of
Botany,
Chinese
Academy
of
Sciences,
Kunming
650204,
Yunnan,
China,
3
Depart-‐
ment
of
Biological
Sciences,
Biochemistry
programme,
Crawford
University,
Igbesa,
Ogun
State,
Nigeria,
4
Department
of
Medical
Biochemistry,
Faculty
of
Basic
Medical
Sciences,
College
of
Medicine,
Lagos
State
University,
Ikeja,
Lagos
State,
Nigeria,
5
Department
of
Biochemistry,
Federal
University
of
Technology,
Akure,
Nigeria
Caesalpinia
bonduc
(family:
Caesalpiniaceae)
has
been
reported
to
have
several
therapeutic
activities
folkloric
and
experimentally
[1,
2].
Two
new
cassane
diterpenoids
1α,7α
-‐diacetoxy-‐
5α,6β-‐dihydroxyl-‐cass-‐14(15)-‐epoxy-‐16,12-‐olide
(1)
and
12α-‐ethoxy-‐1α,14β-‐diacetoxy-‐
2α,5α-‐dihydroxy-‐cass-‐13(15)-‐en-‐16,12-‐olide
(2)
and
known
compounds,
bonducellin
(3)
[3],
7,4'-‐dihydroxy-‐3,11-‐dehydrohomoisoflavanone
(4),
daucosterol
(5),
luteolin
(6),
quercetin-‐3-‐
methyl
ether
(7)
and
kaempferol-‐3-‐O-‐α-‐L-‐rhamnopyranosyl-‐(1
2)-‐β-‐D-‐xylopyranoside
(8)
were
isolated
from
the
ethanolic
extract
of
the
leaves
and
twigs
of
C.
bonduc.
Their
structures
were
elucidated
mainly
by
analysis
of
their
NMR
and
MS
data.
All
compounds
were
evaluated
for
their
antifungal
activity
against
Candida
albicans.
The
antioxidant
properties
of
the
extract
and
compounds
were
assessed
by
the
measurement
of
the
total
phenolic
content,
ascorbic
acid
content,
total
antioxidant
capacity
and
1-‐1-‐diphenyl-‐2-‐picryl
hydrazyl
(DPPH)
scaveng-‐
ing
activity.
Compound
3
showed
moderate
antifungal
activities
with
IC50
value
of
6.45
μg/ml.
Compounds
3,
6,
7
and
ethanolic
extract
had
DPPH
scavenging
activities
with
IC50
values
of
186,
75,
17
and
102
µg/ml
respectively
when
compared
to
vitamin
C
with
IC50
values
of
15
µg/ml.
In
addition,
compound
7
has
the
highest
phenolic
content
of
0.81±0.01
mg/ml
of
Gallic
acid
equivalent
while
compound
8
showed
the
highest
total
antioxidant
capacity
with
254.31±3.54
and
199.82±2.78
µg/ml
Gallic
and
Ascorbic
acid
equivalent
respectively.
All
the
isolated
compounds
and
extract
showed
a
moderate
Ascorbic
acid
content
of
2.26±0.01
to
6.78±0.03
mg/ml.
The
results
obtained
showed
the
antioxidant
activity
of
the
ethanolic
ex-‐
tract
of
C.
bonduc
and
its
extracted
compounds.
Acknowledgement:
We
appreciate
the
Chinese
Academy
of
Sciences
(CAS)
and
the
Academy
of
Sciences
for
the
Developing
World
(TWAS)
for
their
financial
and
material
supports
through
the
CAS-‐TWAS
2009
Post-‐Graduate
Research
Fellowship
granted
to
the
lead
author.
This
work
was
also
supported
by
the
grant
from
the
National
Natural
Science
Foundation
of
China
(30725048)
given
to
Dr.
Ning
H.
Tan
and
also
supported
by
the
Program
for
Promotion
of
Fundamental
Studies
in
Health
Science
of
the
National
Institute
of
Biomedical
innovation
(NIBIO)
(09-‐21,
PI:Yusuke
Wataya).
Keywords:
Cassane
diterpenoids,
Caesalpinia
bonduc,
antifungal
activity,
structural
elucida-‐
tion
References:
[1] Ogunlana
OO,
He
WJ,
Fan
JT,
Zeng
GZ,
Ji
CJ,
Zheng
YQ,
Olagunju
JA,
Akindahunsi
AA,
Tan
NH.
Cytotoxic
flavonoids
from
the
young
twigs
and
leaves
of
Caesalpinia
bonduc
(Linn)
Roxb.
Pak
J
Pharm
Sci
2015;
28:
2191-‐2198
[2] Ogunlana
OO,
Kim
HS,
Wataya
Y,
Olagunju
JO,
Akindahunsi
AA,
Tan
NH.
Antiplasmodial
flavonoid
from
young
twigs
and
leaves
of
Caesalpinia
bonduc
(Linn)
Roxb.
J
Chem
Pharm
Res
2015;
7:
931-‐937
[3] McPherson
DD,
Cordell
GA,
Soejarto
DD,
Pezzuto
JM,
Fong
HHS.
Peltogynoids
and
homoiso-‐
flavonoids
from
Caesalpinia
pulcherrima.
Phytochemistry
1983;
22:
2835-‐2838
P727
Microbial
transformation
of
ruscogenins
by
Cunninghamella
blakesleeana
Özge Özçinar1, Özgür Tağ2, Bijen Kivçak1, Erdal Bedir3
1Department of Pharmacognosy, Ege University, 35100 İzmir, Turkey, 2Bionorm Natural Products Pro-‐
duction
&
Marketing
Co
,İTOB,
35100
İzmir,
Turkey,
3Department
of
Bioengineering,
Izmir
Institue
of
Technology,
35430
İzmir,
Turkey
The
natural
product
drug
discovery
process
involves
the
isolation
of
new
molecules
from
nat-‐
ural
sources,
investigation
of
their
biological
activities,
and
semi-‐synthesis
of
more
active
ana-‐
logs.
Microbial
transformation
plays
a
vital
role
in
the
preparation
of
new
oxygenated
deriva-‐
tives,
and
has
frequently
been
used
as
microbial
model
of
mammalian
drug
metabolism
[1,2].
It
has
been
proved
that
the
hydroxylation
of
steroidal
compounds
is
catalyzed
by
cytochrome
P450
monoxygenase
systems,
which
exist
in
all
eucaryotic
microorganisms
[3].
Cunning-‐
hamella
genus
has
been
widely
used
in
transformation
of
steroids
[4,5].
The
major
steroidal
saponins
of
Ruscus
aculeatus,
ruscogenin
and
neoruscogenin,
has
strong
anti-‐inflammatory
activities,
acts
as
an
anti-‐elastase,
and
decreases
capillary
permeability
[6].
In
the
present
study
microbial
transformation
of
Neoruscogenin:Ruscogenin (78:22) mixture
by
Cunning-‐
hamella
blakesleeana
fungus
afforded
three
new
compounds.
The
structures
were
elucidated
by
LC-‐MS,
1D-‐
and
2D
NMR
analyses
as
shown
below.
Mainly
oxydation
products
were
ob-‐
tained
from
neoruscogenin
by
C.
blakesleana.
As
far
as
can
be
ascertained
from
the
literature,
this
is
the
first
microbial
transformation
study
performed
on
neoruscogenin.
Acknowledgements: We are very grateful to Bionorm Natural Products for providing ruscogenins.
Keywords: Microbial transformation, Ruscogenin, Neoruscogenin, Cunninghamella
blakesleeana
References:
[1] Pazmino
Torres
DE,
Winkler
M,
Glieder
A,
Fraaije
MW.
Monooxygenases
as
biocatalysts:
Classification,
mechanistic
aspects
and
biotechnological
applications.
J
Biotechnol
2010;
146:
9–24
[2] Clark
AM,
Hufford
CD.
Use
of
microorganisms
for
the
study
of
drug
metabolism:
An
update
Med
Res
Rev
1991;
11:
473–501
[3] Ohnishi
T,Yokota
T,
Mizutani
M.
Insights
into
the
function
and
evolution
of
P450s
in
plant
steroid
metabolism
Phytochemistry
2009,
70:
1918–1929
[4] Dong
T,
Wu
GW,
Wang
XN,
Gao
JM,
Chen
JG,
Lee
SS.
Microbiological
transformation
of
dios-‐
genin
by
resting
cells
of
filamentous
fungus,
Cunninghamella
echinulata
CGMCC
3.2716.
J
Mol
Catal.B:
Enzym
2010;
67:
251–256
[5] Hea
X,
Wang
X,
Liu
B,
Su
L,
Wang
G,
Qu
G,
Yao
Z,
Liu
RH,Yao
X.
Microbial
transformation
of
methyl
protodioscin
by
Cunninghamella
elegans.
J
Mol
Catal
B:
Enzym
2005;
35:33–40
[6] Capra
C.
Pharmacology
and
toxicology
of
some
components
of
Ruscus
aculeatus
L.
Fitot-‐
erapia
1972;
43:
99–113
P728
Synthetic
derivatives
of
pulchrol:
An
antiparasitic
natural
prod-‐
uct
Paola Terrazas1,2, Olov Sterner1
1 Center of Analysis and Synthesis, Lund University, Naturvetarvägen 14, SE-‐22100 Lund, 2 Center of
Agroindustrial
Technology,
Mayor
de
San
Simon
University,
La
Torre
Park
and
Sucre
Street,
Cochabamba,
Bolivia.
Natural
products
from
plants
are
one
of
the
most
important
sources
of
new
leading
therapeu-‐
tic
compounds.
In
Mexico
Bourreria
pulchra
(Boraginaceae)
is
used
to
treat
viral
infections,
fever
and
cutaneous
diseases
by
the
people
of
Yucatan
[1].
An
extract
of
its
roots
has
been
previously
studied
and
the
benzochromene
pulchrol
was
isolated
showing
biological
activity
against
Leishmania
mexicana
and
Tripanozoma
cruzi,
parasites
responsible
for
the
neglected
diseases
Leishmania
and
Chagas
[2].
After
its
isolation,
pulchrol
was
synthesized
by
Killander
[3].
In
the
present
study,
options
of
a
total
synthesis
of
pulchrol
derivatives
were
studied
in
order
to
change
the
functional
group’s
position,
the
size
and
nature
on
the
three
rings.
Changes
in
ring
A
such
as
oxidation,
reduction
and
esterification
were
possible
without
diffi-‐
culties;
modifications
on
substituent
position
on
ring
C
were
developed
successfully,
although
an
increase
in
the
number
of
substituents
in
this
ring
was
not
possible;
finally
an
increase
in
the
substituent’s
size
in
ring
B
lead
mainly
to
byproduct
formation.
Thirteen
derivatives
were
obtained
which
have
shown
facile
changes
in
ring
A
and
restricted
modifications
in
ring
B
due
to
its
difficult
subsequent
cyclization.
This
study
is
intended
to
design
of
new
derivatives
of
pulchrol
which
could
improve
the
biological
activity
of
this
compound.
Acknowledgements:
Sofia
Essen
for
her
help
in
the
mass
spectrum
analysis
Keywords: Benzochromene, pulchrol, Bourreria pulchra, synthesis, derivatives.
References:
[1] Argueta
VA,
Cano
LM,
Rodarte
ME.
Atlas
de
las
Plantas
de
la
Medicinal
Tradicional
Mexi-‐
cana.
Mexico:
Instituto
Nacional
Indigenista;
1994:
483-‐485
[2] Erosa-‐Rejón
GJ,
Yam-‐Puc
A,
Chan-‐Bacab
MJ,
Giménez-‐Turbax
A,
Salamanca
E,
Peña-‐
Rodríguez
LM,
Sterner
O.
Benzochromenes
of
the
roots
of
Bourreria
pulchra.
Phytochem
Lett
2010;
3:
9-‐12
[3] Killander
D,
Sterner
O.
Synthesis
of
the
bioactive
benzochromene
pulchrol
and
pulchral,
metabolites
of
Bourreria
pulchra.
Eur.
J.
Org.
Chem
2014;
8:
1594-‐1596
P729
Rational
design
of
annotine
analogues
with
increased
inhibitory
activity
towards
acetylcholinesterase
Elsa
S.
Halldorsdottir1,
Sebastian
Oddsson1,
Arndis
M.
Einarsdottir1,
Borghildur
Eiriksdottir1,
Natalia
M.
Kowal1,
Elin
S.
Olafsdottir1
1
Faculty
of
Pharmaceutical
Sciences,
School
of
Health
Sciences,
University
of
Iceland,
Hagi,
Hofsvallagata
53, IS-‐107 Reykjavik, Iceland
Club
mosses
are
plants
belonging
to
the
family
of
Lycopodiaceae.
They
produce
lycopodium
alkaloids
with
compact,
stable
structures
of
considerable
pharmacological
interest
[1,2].
They
can
interfere
with
important
neurological
targets
for
Alzheimer’s
and
other
neurodegenerative
diseases
and
have
favorable
pharmacological
properties
[2].
Lycodane-‐
type
alkaloids
such
as
huperzine
A
and
B,
are
known
to
be
potent
acetylcholinesterase
(AChE)
inhibitors
[2].
Another
lycopodium
alkaloid,
annotine,
is
a
lycopodane-‐type
alkaloid
found
exclusively
in
Lycopodium
annotinum
[2-‐4].
In
spite
of
fitting
into
the
binding
site
of
the
enzyme,
annotine
creates
only
a
few
weak
interactions
which
make
it
a
very
weak
inhibitor
of
AChE
(IC50
=
860
µM)
[4].
In
this
study
180
analogues
of
annotine
(1)
were
designed
in
silico,
docked
into
a
crystal
structure
of
AChE
(PDB
ID:
1GPN)
and
subsequently
top
score
analogues
were
chosen
for
semi-‐synthesis
with
the
aim
of
obtaining
higher
inhibition
towards
AChE
in
vitro.
The
analogues
annotinol
(2),
O-‐acetyl-‐annotinol
(3),
O-‐benzoyl-‐annotinol
(4),
o-‐fluoro-‐
O-‐benzoyl-‐annotinol
(5)
and
p-‐fluoro-‐O-‐benzoyl-‐annotinol
(6)
were
synthesized
and
tested
for
AChE
inhibition
by
the
colorimetric
method
of
Ellman
[5].
All
analogues
tested
showed
increased
inhibition
of
AChE
as
predicted
by
molecular
modelling
studies;
annotinol
(2)
being
the
most
active
one
showing
about
10
times
the
activity
of
annotine
(1).
Acknowledgements:
Financial
support
from
The
Icelandic
Research
Fund,
The
University
of
Iceland
Re-‐
search
Fund,
The
Icelandic
Research
Fund
for
Graduate
Students,
Bergthora
and
Th.
Scheving
Thorsteins-‐
son
Fund
and
Selma
and
Kay
Langvad
Fund
are
gratefully
acknowledged.
Keywords: Lycopodium alkaloids, annotine, semi-‐synthesis, ligand docking, acetylcholines-‐
terase inhibition
References:
[1] Ma
XQ,
Gang
DR.
The
Lycopodium
alkaloids.
Nat
Prod
Reports
2004;
21:
752-‐772
[2] Olafsdottir
ES,
Halldorsdottir
ES,
Pich
NM,
Omarsdottir
S.
Lycopodium
alkaloids:
Pharma-‐
cology.
In:
Ramawat
K,
Merillon
J,
editors.
Handbook
of
Natural
Products
–
Phytochemis-‐
try,
Botany,
Metabolism,
Vol.
1:
Alkaloids.
Springer-‐Verlag
Berlin
Heidelberg:
2013;
1239-‐
1262
[3] Szarek
WA,
Adams
KAH,
Curcumelli-‐Rodostamo
M,
MacLean
DB.
Lycopodium
alkaloids:
XVI.
Annotine.
Can
J
Chem
1964;
42:
2584-‐2594
[4] Halldorsdottir
ES,
Jaroszewski
JW,
Olafsdottir
ES.
Acetylcholinesterase
inhibitory
activity
of
lycopodane-‐type
alkaloids
from
the
Icelandic
Lycopodium
annotinum
ssp.
alpestre.
Phy-‐
tochem
2010;
71:
149-‐157
[5] Ellman
GL,
Courtney
KD,
Andres
jr
V,
Featherstone
RM.
A
new
and
rapid
colorimetric
de-‐
termination
of
acetylcholinesterase
activity.
Biochem
Pharmacol
1961;
7:
88-‐95
P730
(‒)-‐Bassianolide,
a
cyclodepsipeptide
from
Bombycis
Corpus:
To-‐
tal
synthesis
and
evaluation
of
its
antitumor
activity
Seoung
Rak
Lee,
Seulah
Lee,
Hee
Jeong
Eom,
Hee
Rae
Kang,
Jae
Sik
Yu,
Tae
Kyoung
Lee,
Jiwon
Baek,
Dahae
Lee,
Won
Se
Suh,
Ki
Hyun
Kim
School of Pharmacy, Sungkyunkwan University, Suwon 440-‐746, Republic of Korea
Bombycis
Corpus
is
a
white-‐stiff
silkworm
(Bai
Jiang
Can)
which
is
a
Bombyx
mori
larvae
(silk
moth
larvae,
Bombycidae)
killed
by
infecting
with
the
entomopathogenic
fungus
Beauveria
bassiana.
This
medicinal
insect
has
been
used
as
a
Chinese
traditional
medi-‐
cine
for
an
anticancer
agent
[1].
However,
its
active
ingredients
associated
with
anticancer
effects
and
underlying
mechanisms
remain
unknown.
One
of
the
ingredients
of
Bombycis
Corpus,
(‒)-‐bassianolide
(1),
represents
rich
pharmacophores
with
diverse
biological
activi-‐
ties
including
potential
cytotoxicity
to
various
cancer
cells
[2].
In
this
study,
we
investigated
efficient
total
synthesis
of
(‒)-‐bassianolide
and
evaluated
its
antitumor
activity
in
order
to
support
the
traditional
usage
of
Bombycis
Corpus
as
an
anticancer
agent.
Efficient
total
syn-‐
thesis
of
(‒)-‐bassianolide
was
designed
and
achieved
in
nine
steps,
with
significant
improve-‐
ments
in
the
overall
yield
of
46.8%
(vs.
7.2%
yield
in
previous
synthesis)
[3].
The
cytotoxicity
of
the
(‒)-‐bassianolide
was
evaluated
against
six
human
tumor
cells,
and
the
results
showed
that
the
(‒)-‐bassianolide
displayed
significant
cytotoxicity
against
A549,
SK-‐OV-‐3,
HepG2,
HCT-‐15,
MCF-‐7
and
MDA-‐MB
231
cell
lines
with
IC50
values
of
7.24,
8.44,
15.39,
6.40,
11.42
and
3.98
μg/mL
respectively.
Especially,
(‒)-‐bassianolide
induced
G0/G1
arrest
associated
with
decrease
of
cyclin
A,
D1
and
increase
of
p53,
MDM2,
p21
expression
in
MDA-‐MB
231
cells.
These
results
demonstrate
that
(‒)-‐bassianolide
possesses
antitumor
activities
via
ar-‐
rest
of
the
cell
cycle
in
MDA-‐MB
231
cells
and
the
synthetic
approach
features
an
efficient
and
mild
method
for
the
formation
of
amide
bonds
through
three
inter-‐
and
intramolecular
cou-‐
pling
reactions.
O
O
O N
O O
O
N N
O
O O
N O
O
O
(−)-bassianolide
Acknowledgements:
This
research
was
supported
by
Basic
Science
Research
Program
through
the
Na-‐
tional
Research
Foundation
of
Korea
(NRF)
funded
by
the
Ministry
of
Science,
ICT
&
Future
Planning
(2015R1C1A1A02037383)
Keywords: Total synthesis, (‒)-‐Bassianolide, Cancer, MDA-‐MB 231, Cell cycle
References:
[1] Koo
BS,
An
HG,
Moon
SK,
Lee
YC,
Kim
HM,
Ko
JH,
Kim
CH.
Bombycis
corpus
extract
(BCE)
protects
hippocampal
neurons
against
excitatory
amino
acid-‐induced
neurotoxicity.
Im-‐
munopharm
Immunot
2003;
25:
191−201
[2] Jirakkakul
J,
Punya
J,
Pongpattanakitshote
S,
Paungmoung
P,
Vorapreeda
N,
Tachaleat
A,
Klomnara
C,
Tanticharoen
M,
Cheevadhanarak
S.
Identification
of
the
nonribosomal
pep-‐
tide
synthetase
gene
responsible
for
bassianolide
synthesis
in
wood-‐decaying
fungus
Xylaria
sp.
BCC1067.
Microbiology
2008;
154:
995−1006
[3] Kanaoka
M,
Isogai
A,
Suzuki
A.
Synthesis
of
bassianolide.
Tetrahedron
1977;
46:
4049−4050
P731
The
effect
of
prenylation
on
the
antimicrobial
activity
of
selected
naturally
occurring
furanones
and
pyranones
Serena
Fiorito1,
Francesco
Epifano1,
Vito
Alessandro
Taddeo1,
Salvatore
Genovese1,
Jabrane
Azelmat2,
Daniel
Grenier2
1
Department
of
Pharmacy,
University
“G.
d’Annunzio”
of
Chieti-‐Pescara,
Via
dei
Vestini
31,
66100
Chieti
Scalo
(CH),
Italy,
2
Groupe
de
Recherche
en
Écologie
Buccale,
Faculté
de
médecine
dentaire,
Université
Laval,
2420
de
la
Terrasse,
Québec
(Québec),
Canada
Naturally
occurring
furanones
and
pyranones
and
semisynthetic
derivatives
are
compounds
that
in
recent
years
attracted
the
attention
of
researchers
due
to
their
promising
and
valuable
pharmacological
properties,
mainly
in
terms
of
antibacterial
and
antibiofilm
effects
[1,2].
In
this
poster
communication,
data
on
synthesis
of
six
prenylated
naturally
occurring
and
sem-‐
isynthetic
furanones
and
pyranones
as
well
as
on
the
antimicrobial
activity
of
the
obtained
adducts
will
be
presented.
Selected
parent
products
were
homosotolone
(syn.
abhexone)
1,
sotolone
2,
norfuraneol
3,
among
the
flavour
compounds
of
beer,
red
wine,
and
several
other
beverages,
maltol
4,
typically
present
in
coniferous
plants,
ethylmaltol
5,
a
semisynthetic
de-‐
rivative
of
4,
largely
used
as
sugar-‐like
tasting
additive,
and
finally
kojic
acid
6,
a
pyranone
extracted
from
several
Aspergillus
strains.
The
corresponding
3,3-‐dimethylallyl,
geranyl,
and
farnesyl
ethers
have
been
obtained
by
alkylation
with
the
respective
alkyl
bromide
in
acetone
at
80
°C
in
the
presence
of
dry
K2CO3
as
the
base.
Parent
and
prenylated
compounds
have
been
tested
for
their
in
vitro
growth
inhibitory
activity
against
Gram-‐positive
(Streptococcus
mutans,
Streptococcus
sobrinus)
and
Gram-‐negative
(Porphyromonas
gingivalis,
Fusobacterium
nucleatum,
Aggregatibacter
actinomycetemcomitans)
oral
bacterial
pathogens
as
well
as
a
rep-‐
resentative
pathogenic
fungi
(Candida
albicans).
Preliminary
data
indicated
that
the
most
ef-‐
fective
effects
were
recorded
against
P.
gingivalis
and
F.
nucleatum.
OH HO O
R
O O O
3
1R=H
2 R = CH3
O
R3 R2
4 R1 = CH3, R2 = OH, R3 = H
5 R1 = CH2CH3, R2 = OH, R3 = H
6 R1 = CH2OH, R2 = H, R3 = OH
O R1
Acknowledgements:
Financial
support
to
this
research
from
University
“G.
d’Annunzio”
of
Chieti-‐Pescara
is
gratefully
acknowledged
Keywords: Furanones, Oxyprenylated secondary metabolites, Prenylation, Pyranones
References:
[1] De
Nys
R,
Givskov
M,
Kumar
N,
Kjelleberg
S,
Steinberg
PD.
Furanones.
Prog
Mol
Subcell
Biol
2006;
42:
55-‐86
[2] Lou
J,
Fu
L,
Peng
Y,
Zhou
L.
Metabolites
from
Alternaria
fungi
and
their
bioactivities.
Mole-‐
cules
2013;
18:
5891-‐5935
P732
Preliminary
investigations
on
seleno-‐analogues
of
plant
oxy-‐
prenylated
secondary
metabolites
Serena
Fiorito1,
Francesco
Epifano1,
Vito
Alessandro
Taddeo1,
Salvatore
Genovese1,
Luca
San-‐
cineto2,
Claudio
Santi2
1
Department
of
Pharmacy,
University
“G.
d’Annunzio”
of
Chieti-‐Pescara,
Via
dei
Vestini
31,
66100
Chieti
Scalo
(CH),
Italy,
2
Department
of
Pharmaceutical
Sciences,
University
of
Perugia,
Via
del
Liceo,
Perugia,
Italy.
During
the
last
decades
the
use
of
selenium-‐containing
compounds
as
potential
and
effective
pharmacologically
active
agents
has
been
of
growing
interest
and
to
organoselenium
deriva-‐
tives
can
be
ascribed
a
certain
number
of
biological
activities
[1].
For
example,
ebselen
repre-‐
sent
a
synthetic
selenium-‐containing
product
now
used
in
therapy
able
to
mimic
GPx
activity
[2].
In
this
talk
were
synthesized
four
Se-‐analogues
of
plant
oxyprenylated
benzoic
acid
deriv-‐
atives
1-‐4
starting
from
commercially
available
methyl
esters
of
p-‐aminobenzoic
and
an-‐
thranilic
acids.
They
were
converted
into
diazonium
salts
by
treatment
with
NaNO2
and
HCl
at
0°C
and
made
to
react
in
the
same
reaction
vessel
with
Na2Se2
to
give
diselenides.
In
2-‐
propanol
solution
the
diselenide
derivatives
were
reduced
with
NaBH4
at
r.t.,
and
finally
al-‐
kylated
with
3,3-‐dimethylallyl
bromide
or
geranyl
bromide
to
provide
samples
1-‐4
in
63-‐74%
yields.
The
GPx-‐like
activitiy
of
the
Se-‐prenyl
derivatives
was
assessed
by
1H
NMR
spectrosco-‐
py
using
the
Iwaoka’s
test
[3],
where
the
reduction
of
H2O2
in
CD3OD
was
performed
using
reduced
dithiothreitol
(DTTred).
The
DTTred/DTTox
ratio
was
measured
by
integrating
proton
resonance
for
CH
protons
at
2.63,
2.87,
3.03,
3.67,
and
3.49
ppm
respectively,
as
shown
in
Fig
below
for
compound
3.
COOH COOH
R Se Se R
1 R = CH3 3 R = CH3
2 R = CH2CH2CH=C(CH3)2
4 R = CH2CH2CH=C(CH3)2
Keywords: Glutathione peroxidise activity, oxyprenylated secondary metabolites, seleno-‐
prenylated compounds, semisynthetic derivatives
References
[1] Achibat
H,
Al
Omari
NA,
Messina
F,
Sancineto
L,
Khouili
M,
Santi
C.
Organoselenium
com-‐
pounds
as
phytochemicals
from
the
natural
kingdom.
Nat
Prod
Commun
2015;
10:
1885-‐
1892
[2] Azad
GK,
Tomar
RS.
Ebselen
a
promising
antioxidant
drug:
mechanism
of
action
and
tar-‐
gets
of
biological
pathways.
Mol
Biol
Rep
2014;
41:
4865-‐4879
[3] Santi
C,
Galli
F,
Piroddi
M,
Tidei
C.
Thiols
oxidation
for
the
evaluation
of
Gpx-‐like
activity.
Phosphorus
Sulfur
Silicon
Relat
Elem
2013;
188:
507-‐508
P733
Chemically
engineered
extracts
of
St
John’s
wort
as
sources
of
polyprenylated
acylphloroglucinols
to
prevent
endothelial
dys-‐
function
Maxime
Le
Bot1,
Nina
Corlay1,
Aurore
Michaud1,
Sylvain
Pagie2,
Pascal
Richomme1,
Béatrice
Charreau2,
Séverine
Derbré1
1
EA921
SONAS/SFR4207
QUASAV,
Université
d’Angers,
France.
2
INSERM,
UMR1064,
Nantes,
France.
Graft
rejection
remains
a
serious
concern
in
transplantation
therapy.
As
endothelial
dysfunc-‐
tion
plays
a
prominent
role
in
transplant
rejection,
finding
new
ways
to
prevent
this
process
is
a
huge
challenge.
Recently,
Rouger
et
al.
evidenced
the
significant
anti-‐inflammatory
and
immunomodulatory
properties
of
polyphenolic
compounds
[e.g.
polyprenylated
acylphloroglucinols
(PPAPs)]
from
tropical
Calophyllaceae
and
Clusiaceae
plants
on
endothe-‐
lial
cells
[1,2].
Plants
from
the
Hypericaceae
family
biosynthesize
similar
natural
products
and
some
species
such
as
St
John’s
wort
(Hypericum
perforatum)
are
cultivated
thus
available
in
large
amounts.
To
preserve
biodiversity
and
valorize
medicinal
plants
growing
in
the
Loire
Valley,
the
HYPROTEC
project
attempts
to
I/
improve
access
to
a
library
of
original
and
bioac-‐
tive
polyphenols
by
chemical
modifications
of
H.
perforatum
extracts
[3];
II/
evaluate
their
potential
for
preventing
endothelial
dysfunction.
Preliminary
results
evaluating
the
inhibition
of
VCAM-‐1
expression
on
endothelial
cells
[1]
by
H.
perforatum
roots
and
flowering
tops
ex-‐
tracts
and
their
fractions,
combined
with
a
dereplication
study,
suggested
that
hyperforin
1
and
its
oxidized
forms
could
prevent
inflammation
and,
consequently,
endothelial
dysfunc-‐
tion.
The
present
work
thus
describes
the
synthesis
and
purification
of
a
small
library
of
PPAPs
derivatives
from
the
cyclohexanic
flowering
tops
extract
of
H.
perforatum
(1:
3.0%
m/m)
that
was
subjected
to
various
oxidation
reactions
using
so-‐called
“green
processes”.
Data
were
eventually
supplemented
by
results
from
biological
evaluation.
In
particular,
hy-‐
perforin-‐DCHA
1
(10µM)
strongly
inhibited
TNF-‐induced
VCAM-‐1
(58%),
ICAM-‐1
(34%)
and
E-‐selectin
(54%)
expression
in
comparison
with
the
reference
compound
PDTC
(200
µM).
Acknowledgements:
This
work
was
supported
by
a
grant
funded
by
the
Region
Pays
de
la
Loire
(HYPRO-‐
TEC
project).
Keywords: Endothelial dysfunction, Hypericum perforatum, hyperforin, immunosuppression,
inflammation.
References:
[1] Rouger
C,
Derbré
S,
Charreau
B,
Pabois
A,
Cauchy
T,
Litaudon
M,
Awang
K,
Richomme
P.
Lepidotol
A
from
Mesua
lepidota
inhibits
inflammatory
and
immune
mediators
in
human
endothelial
cells.
J
Nat
Prod
2015;
78:
2187-‐2197.
[2] Rouger
C,
Pagie
S,
Derbré
S,
Ray
A-‐ML,
Richomme
P,
Charreau
B.
Anti-‐inflammatory
and
immunosuppressive
effects
of
polyphenols
from
Clusiaceae
and
Calophyllaceae
on
endothelial
cells.
PloS
One:
submitted
[3] Ayelen
Ramallo
I,
Salazar
MO,
Mendez
L,
Furlan
RLE.
Chemically
engineered
extracts:
Source
of
bioactive
compounds.
Acc
Chem
Res
2011;
44:
241-‐250
P734
Design,
synthesis
and
biological
evaluation
of
novel
emodin
de-‐
rivatives
as
potent
antidyslipidemic
and
antioxidant
agents
Sukanya
Pandeti1,
Ravi
Sonkar2,
Gitika
Bhatia2,
Narender
Tadigoppula1
1Medicinal
and
Process
Chemistry
Division,
CSIR-‐Central
Drug
Research
Institute,
Lucknow-‐226
031,
U.P
India,
2Biochemistry
Division,
CSIR-‐Central
Drug
Research
Institute,
Lucknow-‐226
031,
U.P
India
Hyperlipidemia
is
the
presence
of
abnormal
levels
of
lipids
or
lipoproteins
in
the
blood,
which
contributes
in
the
manifestation
of
atherosclerosis
and
other
cardiovascular
diseases
[1].
As
a
part
of
our
drug
discovery
program
on
antidyslipidemic
agents,
a
large
quantity
of
Emodin
(1)
was
isolated
from
the
roots
of
Rheum
emodi
L.
(Polygonaceae)
[2].
We
studied
antidyslipidem-‐
ic
activity
in
triton
(WR-‐1339)
induced
hyerlipidemic
rat
model
the
naturally
occurring
an-‐
thraquinone
emodin
(1)
and
its
synthetic
derivatives
2-‐15.
The
administration
of
triton
in
rats
induced
marked
hyperlipidemia
by
increasing
the
plasma
levels
of
TC
(2.11
folds),
PL
(2.37
folds)
and
TG
(2.44
folds)
compared
to
control
rats.
Treatment
of
hyperlipidemic
rats
with
2-‐15
at
100
mg/kg
p.o.
reversed
the
plasma
levels
of
lipids.
Emodin
(1)
caused
a
de-‐
crease
in
plasma
levels
of
TC
by
27%
(133.50
±
10.06
mg/dl),
PL
by
26%
(150.75
±
11.08
mg/dl)
and
TG
by
28%
(155.36
±
11.05
mg/dl)
respectively
as
compared
to
triton
induced
rats.
Among
2-‐15,
the
C-‐alkylated
emodin
derivative
9
turned
out
to
be
most
potent
lipid
low-‐
ering
agent,
causing
a
decrease
in
plasma
levels
of
TC,
PL
and
TG
by
39%,
35%
and
39%,
re-‐
spectively,
as
compared
to
triton
induced
rats
whereas
the
marketed
lipid
lowering
drug
gem-‐
fibrozil
decreased
the
levels
of
TC,
PL
and
TG
in
plasma
by
34%,
33%
and
33%,
respectively.
Further
compound
9
was
studied
for
lecithin-‐cholesterol
acyltransferase
(LCAT)
activity.
Ad-‐
ministration
of
triton
in
rats
markedly
decreased
LCAT
activity
in
liver
by
38%.
After
treat-‐
ment
with
compound
9,
LCAT
activity
was
significantly
increased
by
25%
similar
to
standard
drug
gemfibrozil.
The
scavenging
potential
of
9
at
200
µg/ml
was
studied
and
a
significant
decrease
in
superoxide
anions
(36%)
and
hydroxyl
radicals
(31%)
were
observed.
Further-‐
more,
compound
9
at
200
µg/ml
reduced
the
microsomal
lipid
peroxidation
(35%).
Altogeth-‐
er
our
results
suggest
that
9
could
be
a
potential
new
class
of
therapeutic
agent
for
dyslipidemia
treatment.
Acknowledgements:
Sukanya
Pandeti
is
thankful
to
Council
of
Scientific
and
Industrial
Research
(CSIR),
New
Delhi
for
financial
support
and
SAIF,
CDRI
for
spectral
data
Keywords:
Emodin
derivatives,
Rheum
emodi,
LCAT
activity,
antidyslipidemic
activity,
antiox-‐
idant
activity,
LDL
oxidation
References:
[1] Grundy
M.
Cholesterol
and
coronary
heart
disease
a
new
era.
JAMA
1986;
256:
2849-‐2858
[2] Agarwal
K,
Singh
S,
Verma
S,
Kumar
S.
Antifungal
activity
of
anthraquinone
derivatives
from
Rheum
emodi.
J
Ethnopharmacol
2000;
72:
43-‐46
P735
Circular
disulfide-‐rich
peptide
scaffolds
as
anti-‐citrullinated
pep-‐
tide
antibody
inhibitors
Sunithi
Gunasekera1,
Catia
Fernandes-‐Cerqueira2,
Camilla
Eriksson1,
Per-‐Johan
Jakobsson2,
Ulf
Göransson
1
1Division of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, BMC, Box 574, 751
23
Uppsala,
Sweden.
2Rheumatology
Unit,
Department
of
Medicine,
Karolinska
Institutet,
Stockholm,
171
76
Solna,
Sweden
There
is
strong
evidence
suggesting
that
Anti-‐Citrullinated
Protein/Peptide
Antibodies
(AC-‐
PA)
are
specifically
involved
in
the
pathogenesis
of
rheumatoid
arthritis
(RA)
[1].
The
current
work
describes
the
use
of
sunflower
trypsin
inhibitor
1
(SFTI-‐1)
scaffold
[2],
a
14
residue
long
disulfide
stabilized
circular
peptide
for
incorporating
bioactive
epitopes
for
RA
therapy.
We
synthesized
a
series
of
high
affinity
(kd=2
nM),
stable,
cyclic
peptide-‐based
scavengers
of
AC-‐
PA,
by
translocating
an
epitope
derived
from
α
fibrinogen
into
the
SFTI-‐1
scaffold.
The
best
scavenger
blocked
79%
of
aCCP2
IgG
(IC50
of
20
μM),
and
approximately
>90%
of
the
peptide
was
retained
after
five
hours
of
incubation
in
blood.
We
found
that
the
peptide
scavenger
could
serve
as
a
chemical
probe
to
capture
a
specific
subtype
of
ACPA
from
human
serum.
In
summary
we
have
shown
proof-‐of-‐concept
that
antibodies
represent
a
druggable
target,
whose
efficient
inhibition
or
‘blocking’
by
stable
cyclic
peptides
show
great
promise
for
ther-‐
apeutic
and
diagnostic
applications
in
autoimmune
diseases
including
RA.
Keywords: anti-‐citrullinated protein/peptide antibodies (ACPA), cyclic peptide, fibrinogen,
NMR, target, sunflower trypsin inhibitor 1 (SFTI-‐1)
References:
[1] Rantapaa-‐Dahlqvist S, de Jong B A, Berglin E, Hallmans G, Wadell G, Stenlund H, Sundin U,
van Venrooij WJ. Antibodies against cyclic citrullinated peptide and IgA rheumatoid factor
predict the development of rheumatoid arthritis. Arthritis Rheum 2003; 48: 2741-‐2749
[2] Goransson U, Burman R, Gunasekera S, Stromstedt AA, Rosengren KJ. Circular proteins from
plants and fungi. J Biol Chem 2012; 287: 27001-‐27006
P736
Engineering
of
KR-‐12:
A
minimalized
domain
derived
from
hu-‐
man
host
defense
peptide
LL-‐37
into
a
potent
antimicrobial
drug
lead
Taj
Muhammad,
Sunithi
Gunasekera,
Adam
A.
Strömstedt,
Ulf
Göransson
Division
of
Pharmacognosy,
Department
of
Medicinal
Chemistry,
Uppsala
University,
Husargatan
3,
Bio-‐
medical
Centrum,
SE
75123,
Uppsala,
Sweden
The
human
cathelicidin
LL-‐37
is
a
multifunctional
host
defence
molecule
that
mediates
vari-‐
ous
host
responses
including
antimicrobial
action,
chemotaxis,
epithelial
cell
activation,
angi-‐
ogenesis,
and
activation
of
chemokine
secretion
[1].
However,
LL-‐37
encounters
the
generic
problems
common
to
most
linear
counter
parts
such
as
short
biological
half-‐life,
primarily
arising
from
proteolytic
susceptibility.
A
minimalized
bacteriolytic
domain
of
LL-‐37,
referred
to
as
KR-‐12,
has
selective
toxicity
toward
bacteria
[2].
In
our
previous
work,
we
have
per-‐
formed
alanine/lysine
scans
on
KR-‐12
to
identify
critical
residues
for
antimicrobial
activity.
Using
the
insights
obtained
from
the
previous
studies,
we
designed
2nd
generation
cyclic
ana-‐
logs
to
improve
peptide
stability
further
and
reduce
cytotoxicity.
The
stable
peptide
leads
were
engineered
by
using
a
combination
of
peptide
stabilization
approach,
namely
backbone
cyclization,
disulfide
bond
formation
and
dimerization.
The
new
peptide
analogues
showed
more
potent
antimicrobial
activity
against
all
the
tested
strains
(E.
coli,
P.
aeruginosa,
S.
aureus
and
C.
albicans)
than
the
1st
generation
cyclic
analogs.
The
activity
was
equivalent
to
the
par-‐
ent
peptide
LL-‐37. Moreover,
the
new
series
of
peptides
showed
more
potency
on
E.
coli
membrane
permabilization
as
compared
to
the
1st
generation
cyclic
analogs.
Notably,
NMR
analysis
revealed
that
the
2nd
generation
cyclic
analogs
were
unstructured,
presumably
be-‐
cause
backbone
cyclization
restricts
conformational
freedom.
In
addition
to
increasing
our
understanding
of
the
structure
activity
relationship
of
KR-‐12,
this
study
highlights
several
viable
peptide
stabilization
strategies
to
improve
both
potency
and
stability
of
the
enzyme
susceptible
KR-‐12,
which
could
serve
as
a
template
for
novel
antibiotic
development.
Acknowledgements:
Swedish
NMR
Centre
Gothenburg
Keywords: LL-‐37, KR-‐12, dimerization, membrane permabilization, NMR
References:
[1] Patricia MS. The human cathelicidin hCAP18/LL-‐37: A multifunctional peptide involved in
mycobacterial infections. Peptides 2010; 31: 1791-‐1798
[2] Wang G. Structures of human host defense cathelicidin LL-‐37 and its smallest antimicro-‐
bial peptide KR-‐12 in lipid micelles. J Biol Chem 2008; 283: 32637-‐32643
P737
In
silico
identification
and
in
vitro
activity
of
new
pancreatic
li-‐
pase
inhibitors
Tina Buchholz1, Anne Frank2, Gerhard Wolber2, Matthias F. Melzig1
1 Department of Pharmaceutical Biology, Freie Universität Berlin, Königin-‐Luise-‐Str. 2+4, 14195 Berlin,
Germany,
2
Department
of
Medicinal
and
Pharmaceutical
Chemistry,
Freie
Universität
Berlin,
Königin-‐
Luise-‐Str.
2+4,
14195
Berlin
Overweight
and
obesity
are
becoming
some
of
the
greatest
threats
to
global
health
in
the
21st
century.
An
important
strategy
in
their
treatment
includes
the
reduction
of
intestinal
fat
ab-‐
sorption
through
inhibition
of
pancreatic
lipase
[1]
(PL,
pancreatic
triacylglycerol
lipase
(EC:
3.1.1.3)),
which
is
the
main
enzyme
that
breaks
down
dietary
fats
in
the
human
digestive
sys-‐
tem
[2,3].
Some
of
the
most
widely
studied
materials
for
natural
PL
inhibitors
are
secondary
metabolites,
of
which
polyphenols,
saponins,
and
terpenes
showed
significant
potential
[4].
In
the
present
study,
polyphenolic
compounds
were
screened
for
PL
inhibitory
activity
by
using
an
enzymatic
in
vitro
assay
based
on
the
hydrolysis
kinetic
of
an
oleate
ester
of
4-‐
methylumbelliferone.
The
polyphenolic
compounds
which
showed
an
anti-‐lipase
effect
were
further
investigated
for
their
binding
site
and
pharmacophore
using
in
silico
computational
modelling
experiments.
By
using
available
plant
resources
data
banks,
the
most
likely
poten-‐
tial
natural
structures
(hits)
were
derived.
In
a
set
of
1699
structures
chosen
for
the
screen-‐
ing,
68
were
predicted
to
possess
inhibitory
activity
towards
the
enzyme.
Eight
candidates
were
therefore
tested
in
the
in
vitro
enzyme
assay
used
before.
The
results
show
that
7
out
of
8
tested
substances
were
able
to
inhibit
PL
in
a
dose-‐dependent
manner
(best
IC50
value:
87.2
±
3.1
µM).
Keywords: Anti-‐obesity, pancreatic lipase inhibition, in silico modelling, polyphenols
References:
[1] Foster-‐Schubert
KE,
Cummings
DE.
Emerging
Therapeutic
Strategies
for
Obesity.
Endocr
Rev
2006;
27:
779–793
[2] Winkler
FK,
D’Arcy
A,
Hunziker
W.
Structure
of
human
pancreatic
lipase.
Nature
1990;
343:
771–774
[3] Kimura
H,
Futami
Y,
Tarui
S,
Shinomiya
T.
Activation
of
Human
Pancreatic
Lipase
Activity
by
Calcium
and
Bile
Salts.
J
Biochem
1982;
92:
243–251
[4] Birari
RB,
Bhutani
KK.
Pancreatic
lipase
inhibitors
from
natural
sources:
unexplored
po-‐
tential.
Drug
Discovery
Today
2007;
12:
879–889
P738
Anti-‐HIV
natural
products
(28):
preparation
of
conjugate
for
3-‐O-‐
acyl
betulin
derivative
and
AZT
as
anti-‐HIV
agents
Shizuka
Wada1,
Naonobu
Tanaka1,
Chin-‐Ho
Chen2,
Susan
L.
Morris-‐Natschke3,
Kuo-‐Hsiung
Lee3,
Yoshiki
Kashiwada1
1
Graduate
School
of
Pharmaceutical
Sciences,
Tokushima
University,
Tokushima
770-‐8505,
Japan,
2
Med-‐
ical
Center,
Duke
University,
Durham,
NC
27710,
USA,
3
Natural
Products
Laboratory,
School
of
Pharmacy,
University
of
North
Carolina,
Chapel
Hill,
NC
27599,
USA
Bevirimat (3-‐O-‐(3',3'-‐dimethylsuccinyl)-‐betulinic acid) was discovered in our modification
study on betulinic acid, which was identified as an anti-‐HIV component from the leaves of
Syzygium claviflorum (Roxb.) Wall. ex Steud. (Myrtaceae) [1,2]. Bevirimat was shown to be the
first-‐in-‐class HIV maturation inhibitor, which disrupts the processing of capside precursor
p25 (CA-‐SP1) to mature capsid protein p24 (CA) [3]. Based on this finding, various bevirimat-‐
related betulin derivatives were prepared and their anti-‐HIV activity evaluated. Among oth-‐
ers, 3-‐O-‐(3',3'-‐dimethylsuccinyl)-‐28-‐O-‐(2",2"-‐dimethylsuccinyl)-‐betulin and 3-‐O-‐glutaryl-‐
dihydrobetulin demonstrated potent anti-‐HIV activity with EC50 values of 8.7 × 10–4 and 2.0 ×
10–5 µM, respectively, and TI values of 4.24 × 104 and 1.12 × 106, respectively [3,4]. In a con-‐
tinuing study on bevirimat-‐related derivatives as anti-‐HIV agents, novel conjugates of 3,28-‐di-‐
O-‐acyl betulin derivative with AZT were also prepared as anti-‐HIV agents based on the strate-‐
gy of ‘double-‐drug’ in drug design. Some of the prepared conjugates exhibited potent anti-‐HIV
activity at the submicromolar level equipotent or more potent than bevirimat [5]. However,
further experiment of the most potent derivative suggested that the conjugate could not split
into two fragments (a betulin derivative and AZT) inside of the lymphocytes under the exper-‐
imental condition. Based on these findings, we modified the design of the conjugate where an
amino acid moiety was introduced in its linker moiety anticipating easily to be hydrolysed by
esterase. Preparation of these conjugates and evaluation of their anti-‐HIV activities will be
presented.
Keywords: Bevirimat, betulin, AZT, anti-‐HIV
References:
[1] Fujioka T, Kashiwada Y, Kilkuskie RE, Cosentino LM, Ballas LM, Jiang JB, Janzen WP, Chen IS,
Lee KH. Anti-‐AIDS agents. 11. Betulinic acid and platanic acid as anti-‐HIV principles from
Syzygiun claviflorum, and the anti-‐HIV activity of structurally related triterpenoids. J Nat
Prod 1994; 57: 243-‐247
[2] Kashiwada Y, Hashimoto F, Cosentino LM, Chen CH, Garrett PE, Lee KH. Betulinic acid and
dihydrobetulinic acid derivatives as potent anti-‐HIV agents. J Med Chem 1996; 39: 1016
[3] Li F, Goila-‐Gaur R, Salzwedel K, Kilgore NR, Reddick M, Matallana C, Castillo A, Zoumplis D,
Marin DE, Orenstein JM, Allaway GP, Freed EO, Wild CT. PA-‐457: A potent HIV inhibitor
that disrupts core condensation by targeting a late step in Gag processing. Proc Natl Acad
Sci USA 2003; 100: 13555-‐13560
[4] Kashiwada Y, Chiyo J, Ikeshiro Y, Nagao T, Okabe H, Cosentino LM, Fowke K, Lee KH. 3,28-‐
Di-‐O-‐(dimethylsuccinyl)-‐betulin isomers as anti-‐HIV agents. Bioorg Med Chem Lett 2001;
11: 183-‐185
[5] Kashiwada Y, Sekiya M, Ikeshiro Y, Fujioka T, Kilgore NR, Wild CT, Allaway GP, Lee KH. 3-‐O-‐
Glutaryl-‐dihydrobetulin and related monoacyl derivatives as potent anti-‐HIV agents.
Bioorg Med Chem Lett 2004; 14: 5851-‐5853
P739
Exploring
natural
coumarin
derivatives
as
agents
against
Myco-‐
bacterial
biofilm,
highlighting
inhibition
and
eradication
Carel
Basson
Oosthuizen1,
Navneet
Kishore1,
Namrita
Lall1
1
Department
of
integrated
plant
and
soil
sciences,
University
of
Pretoria,
Plant
Science
Complex,
C/o
Lynnwood
road
&
Herold
street,
Pretoria,
South
Africa.
Coumarin,
a
benzo-‐α-‐pyrone
class
compound,
was
first
isolated
from
Dipteryx
odorata.
Cou-‐
marins
are
biologically
active
compounds
found
in
several
plant
families
[1].
Tuberculosis
remains
a
burden
in
many
countries
in
the
world.
It
is
believed
that
the
Tuberculosis
bacteria
utilize
mechanisms
to
evade
chemotherapeutic
attack
and
immune
responses
[2].
This
in-‐
cludes
a
quorum
sensing
regulated
biofilm
growth
habit
[3].
In
this
study
two
coumarin
base
compounds;
7-‐hydroxy-‐umbeliferone
and
4-‐methyl-‐umbeliferone
were
used
to
derivatize
12
compounds.
They
were
tested
for
multiple
activities
related
to
Mycobacterial
infections,
with
the
main
focus
on
biofilms.
Antimycobacterial
and
antibiofilm
assays
were
conducted
to
de-‐
termine
their
efficacy.
It
was
found
that
the
antibacterial
activity
was
medium
to
low
with
minimum
inhibitory
concentrations
(MIC’s)
ranging
between
50
µg/ml
(Samples
U1,
U2)
and
100
µg/ml
(Samples
U3,
U4).
Although
this
seems
low,
the
derivatives
appear
to
be
selective
towards
biofilm
inhibition
with
a
selective
index
(SI
=
Antimycobacterial
antivity/Antibiofilm
activity)
of
60
for
U1,
U2,
MU1
and
MU2
suggesting
the
importance
of
the
moieties
attached.
Due
to
the
hepatotoxicity
related
to
the
TB
regimen,
the
samples
were
tested
for
their
hepa-‐
toprotective
vs.
hepatotoxic
activity
on
HepG2
hepatocytes,
with
a
toxicity
induced
by
aceta-‐
minophen.
Samples
U1-‐U5
and
MU1-‐MU4
showed
40%
and
20%
protection
respectively.
All
the
samples
showed
low
to
no
toxicity
with
sample
U2
showing
the
highest
with
an
IC50
of
46.52
µg/ml.
These
coumarin
derivatives
showed
promising
results
for
biofilm
inhibition
and
should
be
considered
for
further
testing
on
different
biofilm
structures.
Acknowledgements:
Medical
Research
Council
SA,
National
Research
Foundation
Keywords: Coumarin, Mycobacterium tuberculosis, biofilm, hepatoprotection.
References:
[1] Tandon S, Rastogi RP. Recent Advances in naturally occurring coumarins. J Sci Ind Res 1979;
38: 428–441.
[2] Ojha AK, Hatfull GF. Biofilms of Mycobacterium tuberculosis: New perspectives of an old
pathogen. Underst Tuberc - Deciphering Secret Life Bacill 2012; 182–192.
[3] Ojha
AK,
Baughn
AD,
Sambandan
D,
Hsu
T,
Trivelli
X,
Guerardel
Y,
Alahari
A,
Kremer
L,
Jacobs
WR,
Hatfull
GF.
Growth
of
Mycobacterium
tuberculosis
biofilms
containing
free
mycolic
acids
and
harbouring
drug-‐tolerant
bacteria.
Mol
Microbiol
2008;
69:
164–174.
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