RECERCA EN PRODUCTES NATURALS Amaryllidaceae com a...
Transcript of RECERCA EN PRODUCTES NATURALS Amaryllidaceae com a...
RECERCA EN PRODUCTES NATURALS
Dr. Laura Torras Claveria; Luciana Tallini, Dr. Francesc
Viladomat, Dr. JaumeBastida
Seminarisde Recercade la Facultatde Farmcia
27 de setembrede 2016
Plantes ornamentals de la famlia Amaryllidaceaecom a font de
compostos bioactius
Hymenocallis littoralis
Phaedranasadubia
Crinum x powellii
Galanthus nivalis
Lycoris radiata
Narcissuscv. Pinza
AMARYLLIDACEAE
1981-2010
Newman & Cragg, J Nat Prod 2007, 70, 461-477
Newman & Cragg, J Nat Prod 2012, 75, 311-335
Natural Products
All newapproveddrugs
Natural product and/or natural product structures play a highly significant role in the drug discovery and development
New drugs Pre-clinic Clinic assays (Phases I-III)FDA Revision
Production/ Phase IV
5 Years 1,5 Years 6 Years 2 Years 2 Years
Pre
sen
taci
n
ND
A
Pre
sen
taci
n
IN
D
5 compounds
Average time in each step
250 compunds
IND: InvestigationalNew Drug
NDA: New DrugApplication
Research and development
of new products
1 approved drugFDA
10.000compounds
AmaryllidaceaefamilyBulbous plant family with ornamental and medical value
3 subfamilies: Agapanthoideae, Allioideae, and Amaryllidoideae(previously 3 families)
Term Amaryllidaceae
corresponds with subfamily Amaryllidoideae
Hippocrates of Kos (460-370 BC). Narcissus oil
Uterus tumors
Dioscorides(40-90 AD)Soranusof Ephesus (98-138 AD)
Same therapy
Pliny the Elder (23-79 AD)Topic use
Narcissus extracts
BibleNarcissus tazetta
Anti-tumor therapy
Middle AgesChinese medicine, N Africa
Narcissus oil
European doctorsHymenocalliscaribaeatumors inflammation
Amaryllidaceaemedical uses through History
Amaryllidaceaegeographical distribution
Amaryllidaceae: Africa
Amaryllidaceae: America
Hymenocalliscoronaria(Alabama)
Amaryllidaceae: Asia
Amaryllidaceae: EuropeMediterranean area
Caucasus
Narcissus poeticus(Vall , Catalonia)
Amaryllidaceaealkaloids
6
4a
1
4a
10b10a
10
9
8
7 6
12
11
4
3
2
norbelladine lycorine homolycorine
crinine narciclasinehaemanthamine
tazettine
montanine
6a'
43
2
16
5
1'2'
3'
4' 6'5'
12 11
3
21
4
4a
10b10a
6a7
109
8
12
3
4
4a10b
10a
6a 6
10
9
87
1
11
12
32
66a
10a10
7
8
910b
4
7 6
11
4
1
2
10b
4a
10a
6a
3
8
910
1
4
4a12
11
6
6a7
10
8
911a 2
3
10a
1210a
6a7
8
910
11 12
1
23
4
4a
10b
6
galanthamine
NH
HO
HO
HO
N
O
O
OH
HO
H
H
O
MeN
MeO
MeO
H
H
O
H
NH
OH
O
O
O
NMe
OMe
H
OH
O
O
4a
1
11
12
32
66a
10a10
7
8
910b
4
NH
OMe
O
O
OH
NH
OH
OH
OH
H
OH O
O
O
N
OMe
OHO
O H
O
NMe
MeO
OH
Other Amaryllidaceaealkaloids
O
NMeO
O
4a
10b
GracilineEsqueleto dibenzo[b,d]piranoConfiguracin absoluta 4aR/10bR
RR
Galanthus gracilisGalanthus plicatus ssp byzantinusPancratium maritimum
N
NMeO
O
OMe
H
O
O
OH
H
PlicamineCompuesto dinitrogenado no dimricoCentro spiro = Tazettina [O/N]
Galanthus plicatus ssp byzantinusCyrthantus obliquus
Graciline Plicamine
Dibenzo[b,d] piranskeletonAbsolute configuration 4aR/10bR
GalanthusgracilisGalanthusplicatusssp. ByzantinusPancratiummaritimum
Non dimeric dinitrogenatedcompoundSpiro center = Tazettine[O/N]
Galanthusplicatusssp. ByzantinusCyrthantusobliquusNarcissus broussonetii
Exceptions
NMe
O
O
O
O
O
H
H O
NMe
O
OH
HO
MeO
NMe2
ONMe2
O NMe
OMe
O
OMe
NMe
OMe
OH
OMe
(-)-capnoidineFumariaceae
(+)-bulbocapninePapaveraceae, Fumariaceae yLauraceae
amisineAizoaceaeHymenocallis arenicolaZ Chem 1980, 20: 298
mesembrenolAizoaceaeCrinum oliganthumZ Chem 1981, 21: 358
mesembrenoneAizoaceaeNarcissus pallidulusJ Nat Prod 1989, 52: 478
NH
HO
MeO
salsolineChenopodiaceae (Salsola richteri)Galanthus woronowii
Me
Galanthus nivalis ssp cilicicusBiochem Syst Ecol 2004, 32, 1059
Biochem Syst Ecol 2013, 51, 276
Chemotaxonomic value
O
NMe
O
NMe
COO
NMe
COOH
Shihunine
Dehidroshihunine
H
BehriatenuifloraGreene
Amaryllidaceae AlliaceaeBastida, Sells, Codina, Viladomat, Len de la LuzPlanta Medica 1996, 62, 575-577
Amaryllidaceaealkaloids biosynthesis
trans-cinnamic acid, R1=R2=Hpara-coumaric acid, R1=OH, R2=Hcaffeic acid, R1=R2=OH
tyramine
Tyr-decarboxylase
protocatechuic aldehyde
PAL
L-TyrL-Phe
HO
H2N COOH
HO
H2N
COOH
R1
R2 CHO
HO
HO
NH2
COOH
1.- Enzimaticpreparation of precursors
-oxidation
PAL
hydroxylases
reduction
Tyr-decarboxylase
CHO
HO
HO
HO
H2N
N
HO
HONH
HO
O
H
N
HOHOHO
HO
OH
H
NH
HO
HO
HO
tyramineprotocatechuic aldehyde
Schiff's base(isomeric structures in solution)
norbelladine
Amaryllidaceaealkaloids biosynthesis2.- Primary ciclation
Amaryllidaceaealkaloids biosynthesis
3.- Enzimaticpreparation of intermediary agents
MeO
NHHO
4'
HO
HO
NHHO
4'
HO
norbelladine O-methylnorbelladine
Amaryllidaceaealkaloids biosynthesis
4.- Secondary ciclation. Diversification
types
galanthamine
types
crinine haemanthaminetazettinenarciclasinemontanine
types
lycorinehomolycorine
ortho-para' para-para' para-ortho'
O-methylnorbelladine
MeO
NHHO
4'
HO
Amaryllidaceaealkaloids biosynthesis
4.- Secondary ciclation. Diversification
types
galanthamine
types
crinine haemanthaminetazettinenarciclasinemontanine
types
lycorinehomolycorine
ortho-para' para-para' para-ortho'
O-methylnorbelladine
MeO
NHHO
OH
Amaryllidaceaealkaloids biosynthesis
4.- Secondary ciclation. Diversification
types
galanthamine
types
crinine haemanthaminetazettinenarciclasinemontanine
types
lycorinehomolycorine
ortho-para' para-para' para-ortho'
O-methylnorbelladine
MeO
NH
OHOH
Natural Products Research
SCREENING STRUCTURAL CHARACTERIZATION
Studyof genusNarcissus
[Spain-Portugal]
Studyof Amaryllidaceae
spp. usedin TradMed[SA/Ibero-America]
Studyof galanthamine
producerspecies[Chemotaxonomy]
Catalan flora
19831980 20111993 2005I II III
Phytochemical screening
SCREENING
Catalan flora
19831980
Collection, Drying, StorageTaxonomic DeterminationObtaining genus informationSimple method rapid selection
NarcissusassoanusDuf.=N. requieniiRoem.
=N. juncifoliusLag.
(Amaryllidaceae)
> 1000 samples Green plants & Fungus
different habitats from Catalonia
Natural Products Research
SCREENING STRUCTURAL CHARACTERIZATION
Studyof genusNarcissus
[Spain-Portugal]
Studyof Amaryllidaceae
spp. usedin TradMed[SA/Ibero-America]
Studyof galanthamine
producerspecies[Chemotaxonomy]
Catalan flora
19831980 20111993 2005I II III
Selection of genus Narcissus
High content in exclusive AmaryllidaceaealkaloidsBiological and pharmacological activity of extracts and alkaloids previously isolated from this family speciesPoor studied genusInfluence area: Mediterranean (easy to work)
In base of:
Selection of genus Narcissus
Distribution South Europe (Iberian Peninsula and Italy) (also in North Africa and East Mediterranean)
80-100 speciesNarko(=narcotic) penetrating smell of flowers from some speciesGrowing from lowlands to highlandsFlowering at the end of winter - spring
*
*
**
Plant material
Extracts FractionsPure
compounds
SpectroscopyUV, IR, MS,
NMR (1D/2D), X Ray
Structure elucidation
Methodology
Alkaloids from genus Narcissus
N
OH
HO
O
OH
H
12
3
4
4a
6
11
12
7
10 10b
6a8
9 10a
Lycorine, NarcissuspseudonarcissusGerrad, 1877: isolationNagakawaet al., 1956: characterization
New alkaloids from Narcissus
N
HO
MeO
H
H
OR2
R1O
1-O-acetylpseudolycorine R1= Ac, R2= H2-O-acetylpseudolycorine R1= H, R2= Ac
N
MeO
MeO
oxoassoanine
O
O
R1O
R2O
MeN
H
H
O
9
9-O-demethylhomolycorine R1=H, R2= Me, R3= Hdubiusine R1= COCH2CHOHMe, R2= Me, R3= OAc9-O-demethyl-2 -hydroxyhomolycorine R1= H, R2= Me, R3= OH8-O-acetylhomolycorine R1= Me, R2= Ac, R3= H
H
NMe
O
O
bicolorine
R3
N
MeO
MeO
vasconine R= Htortuosine R= OMe
N
MeO
MeO
OMe
roserine
O
R1O
R2O
MeN
H
H9
H
OMe
O-methyllycorenine R1=R2= Me, R3=H2 -hydroxy-6-O-methyloduline R1+R2= CH2, R3=OH
R3
R
New alkaloids from Narcissus
OMeO
OH
N
H
O
N-formylnorgalanthamine
N
OH
HO
MeO
9
H
9-O-demethylmartidine
OMeO
NH
epinorgalanthamine
OMeO
NH
epinorlycoramine
OH OH
N
OAc
HO
MeO
9
H
cantabricine
N
OMe
H
11-O-acetylhaemanthamine
N
OMe
H
bujeine
OAcO
O
O
O
MeCOOCH2
O
nangustine
O
O N H OH
OH
O
NH
HO H
O
O
obesine
O
NMeO
O
OMe
OOMe
N
HO
pallidiflorine
11
Bastida, Lavilla, Viladomat. TheAlkaloids2006, 63, 87-179
Natural Products Research
SCREENING STRUCTURAL CHARACTERIZATION
Studyof genusNarcissus
[Spain-Portugal]
Studyof Amaryllidaceae
spp. usedin TradMed[SA/Ibero-America]
Studyof galanthamine
producerspecies[Chemotaxonomy]
Catalan flora
19831980 20111993 2005I II III
Amaryllidaceaein traditional medicine
KwaZulu-Natal200.000 Traditionalhealers(Year2004 licencesauthorization) South Africa
Amaryllidaceaein traditional medicine
Brunsvigia
B. josephinaeB. orientalisB. radulosaB. littoralisB. bosmeniae
CrinumC. sthulmaniiC. kirkiiC. delagoenseC. macowanii
CyrtanthusC. elatusC.sanguineusC. obliquus
Nerine
N. filifolia
stomach painLeprosy affected areas
pregnancy (facilitate birth)
sexually transmitted diseases (STD)lactation
urinary problems
South Africa
Amaryllidaceaein traditional medicine
AmmocharisA. tinneana
BoophaneB. flava
Used by indigenous population (Xhosa, Shoto, San, Zulu ...) Infusions of bulbs [> 200 years]. A lot of species from genus Boophane, Brunsvigia, Crinum andNerineare used to treat infertility
South Africa
Plant material
Extracts FractionsPure
compounds
Biological assays
Mass spectrometryGC-MS, LC-MS
SpectroscopyUV, IR, MS,
NMR (1D/2D), X Ray
Structure elucidation
Methodology
Amaryllidaceaealkaloids
South Africa
N
OH
H
MeO
HO
N
OAc
OH
R1
R2
H
N
O
O
OH
OMe
OMe
HN
O
O
OH
OMe
OMe
H
N
OR2
R1O
R3
O
OH
N
O
O
OAc
HO
H
N
O
O
OMe
AcO
OMe
HN
O
O
OMe
O
OMe
H
N
O
O
O NH
OOR1
OHOMe
josephinine
11-O-acetylambelline
brunsbelline
1-epibowdensine R1= R2= Ac, R3= OMe1-epidemethoxybowdensine R1= R2= Ac, R3= H1-epideacetylbowdensine R1= R2 =H, R3= OMe
1
delagoensine R1+R2= OCH2Odelagoenine R1= R2= OMe
6 -hydroxycrinamidine R= H6 -hydroxyundulatine R= Me
macowiine 6 -hydroxybuphanidrine
filifoline
OH
Amaryllidaceaealkaloids
South Africa
N
MeO
HO
H
H
HO
NH
H
AcO
O
O
OAc
O
O
O
HN
O
N
NMeO
O
OMe
O
H
H
OH
N
O
O
OMe
OH
NMe
MeO
RO
N
MeO
HO
H
H
AcO
NH
MeO
MeO
MeO
O
O
O
N
O
kirkine
buflavine R= Me8-O-demethylbuflavine R= H
8
montabuphine
1-O-acetylnorpluviine
obliquine
1,2-O-diacetylzephyranthine
noraugustamine 4a-dedihydronoraugustamine
N-demethylbelladine
Amaryllidaceaein Ibero-America
Amaryllidaceaealkaloids
Ibero-America
N
O
O
O N
MeO
MeO
O
H
N
MeO
MeO
OMe O
NH
O
O
O
O
NMeO
O
OMe
OMe
H
H
ungeremine oxomaritidine
7-methoxyassoanine
apohaemanthamine
6-O-methylpretazettine
MeO
O
NMe
OH
OH
11 -hydroxygalanthamine
Andrade, Berkov, Viladomat, Codina, Zuanazzi, BastidaMolecules2011,16, 7097-7104 (11 -hydroxygalanthamine)
Amaryllidaceaemicropropagation
Ibero-America
Study of species in danger of extinctionStudy of insufficient samples
Natural Products Research
SCREENING STRUCTURAL CHARACTERIZATION
Studyof genusNarcissus
[Spain-Portugal]
Studyof Amaryllidaceae
spp. usedin TradMed[SA/Ibero-America]
Studyof galanthamine
producerspecies[Chemotaxonomy]
Catalan flora
19831980 20111993 2005I II III
Galanthamine
Galanthamine
H3CO
NCH3
O
OH Acetylcholinesteraseinhibitory activity
Interaction with nicotinic receptors
Commercialized for the treatment of mild to
Geographic study Leucojumaestivum
Distribution of different alkaloid types in Leucojumaestivumpopulations in Bulgaria
Berkov, Georgieva, Kondakova, Viladomat, Bastida, Atanassov, CodinaBiochemical Systematics in Ecology 2013, 46, 152-161
Methodology: GC-MS
software AMDISAutomaticMassSpectralDeconvolutionand IdentificationSystem
Separationand identificationof Amaryllidaceaealkaloids in complex mixtures and extracts without derivatisating
Structural information of new compounds
Differentiation of coelutingcompounds
Methodology: GC-MS
software AMDISAutomaticMassSpectralDeconvolutionand IdentificationSystem
Identification of Amaryllidaceaealkaloids
Amaryllidaceaealkaloids library NIST databaseLiterature
KovatsRetention IndexMass spectra
Methodology: GC-MSsoftware AMDIS
GAL in ornamental Narcissus
105 ornamental Narcissuscultivars were evaluated in terms of GAL content and AChEinhibitory activity, as a new source of GAL for pharmaceutical industry
Torras-Claveria, Berkov, Codina, Viladomat, Bastida Industrial Cropsand Products, 2013, 43, 237-244
Narcissus hispanicus
GAL in ornamental Narcissus
GAL in ornamental Narcissus
GAL in ornamental Narcissus
Bulbs and leaves were evaluated separately
Torras-Claveria, Berkov, Codina, Viladomat, Bastida Industrial Cropsand Products, 2013, 43, 237-244
GAL contentGC-MS
AChEinhibitory activity
In vitro test
GAL content
Torras-Claveria, Berkov, Codina, Viladomat, Bastida Industrial Cropsand Products, 2013, 43, 237-244
0
1000
2000
3000
4000
5000
6000
7000
8000
N. confusus N. hispanicusYellow WingsRijnveld EarlySensation
Rip VanWinkle
g
Ga
l /g
DW
Galanthamine
content in
leaves
0
2000
4000
6000
8000
10000
12000
14000
16000
N. confususYellow WingsBella Estrella Enkele JonquillaSimplex
N. hispanicus
g
Ga
l/g D
W
Galanthamine
content in
bulbs
AchEinhibitory activity
Torras-Claveria, Berkov, Codina, Viladomat, Bastida Industrial Cropsand Products, 2013, 43, 237-244
0,000
0,200
0,400
0,600
0,800
1,000
1,200
Snowball GreenHoward
N. confusus Hawera Albus PlenusOdoratus
PinkParadise
N. hispanicus
AC
hEi
nh
ib. A
ct.(
IC50)
0,000
0,200
0,400
0,600
0,800
1,000
1,200
N. confusus Hawera Little Spell
AC
hEi
nh
ib. A
ct.
(IC
50)
IC 50
AChE
inhibition
in leaves
IC 50
AChE
inhibition
in bulbs
AchEinhibitory activity
Torras-Claveria, Berkov, Codina, Viladomat, Bastida Industrial Cropsand Products, 2013, 43, 237-244
Galanthaminecontent
AChEactivity inhibition
N. hispanicus
Cluster analysis Clustering, the organization of a collection of patterns based on similarity. Its aim to find structure in data
LEAFsamples
ALKALOID PROFILE
BULBsamples
5 clusters
5 clusters
Torras-Claveria, Berkov, Codina, Viladomat, Bastida Industrial Cropsand Products, 2014, 56, 211-222
Hierarchical cluster
RupSud Vel
LEAFsamples
BULBsamples
Torras-Claveria, Berkov, Codina, Viladomat, Bastida Industrial Cropsand Products, 2014, 56, 211-222
Lycorine type
0,00
500,00
1000,00
1500,00
2000,00
2500,00
3000,00
D1 D2 D3 D4 D5 D6 D7 D8 D9 D10 D11 D12 D13
g
GA
L/D
W
Horticultural divisions
Galanthamine type
Haemanthamine type
Homolycorine type
Lycorine type
Narciclasine type
Tazettine type
Other
Protoalkaloids
LEAFsamples Horticultural divisions
Galanthamine type
0,00
500,00
1000,00
1500,00
2000,00
2500,00
3000,00
D1 D2 D3 D4 D5 D6 D7 D8 D9 D10 D11 D12 D13
g
GA
L/D
W
Horticultural divisions
Galanthamine type
Haemanthamine type
Homolycorine type
Lycorine type
Narciclasine type
Tazettine type
Other
Protoalkaloids
BULBsamples Horticultural divisions
Statistical analysis
Correspondence analysis
C1 C2 C3 C4 C5 All
D1 0 7 1 8 0 16
D2 0 4 0 9 0 13
D3 0 3 0 9 0 12
D4 0 4 0 8 0 12
D5 0 2 0 5 0 7
D6 0 7 2 6 0 15
D7 0 2 0 6 0 8
D8 0 0 1 0 0 1
D9 0 2 0 0 0 2
D10 0 1 0 2 0 3
D11 0 3 0 3 0 6
D12 0 2 0 0 0 2
D13 1 1 2 4 1 9
All 1 38 6 60 1 106
C1 C2 C3 C4 C5 All
D1 4 2 3 7 0 16
D2 2 3 3 5 0 13
D3 2 5 3 2 0 12
D4 7 1 3 1 0 12
D5 1 0 6 0 0 7
D6 6 6 2 1 0 15
D7 2 1 3 2 0 8
D8 0 0 1 0 0 1
D9 0 1 0 1 0 2
D10 0 0 3 0 0 3
D11 1 0 1 4 0 6
D12 0 1 0 1 0 2
D13 2 0 2 4 1 9
All 1 38 6 60 1 106
Cross tabulation
Torras-Claveria, Berkov, Codina, Viladomat, Bastida Industrial Cropsand Products, 2014, 56, 211-222
Natural Products Research
Metabolomics
Ibero-America
Lycoris
2011 - today
Metabolomics
Torras-Claveria, Berkov, Juregui, Caujap, Viladomat, Codina, Bastida Phytochemicalanalysis, 2010, 21, 80-88
Pancratiumcanariense
Pigni, Berkov, Ros-Ruiz, Luque, Viladomat, Codina, Bastida Phytochemistry, 2013, 95, 384-393
Narcissus SectionGanymedes
Torras-Claveria, Berkov, Codina, Viladomat, BastidaIndustrial CropsandProducts, 2014, 56, 211-222
OrnamentalNarcissus varieties and cultivars
Metabolomics
Parejo, Juregui, Viladomat, Bastida, Codina. RapidCommunications in MassSpectrometry, 2004, 18, 2801-2810
Polyphenols LC-MS/MS and LC high resolution MS2000 - today
Snchez-Rabaneda, Juregui, Lamuela-Ravents, Bastida, Viladomat, Codina. Journalof ChromatographyA, 2003, 1008, 57-72
Snchez-Rabaneda, Juregui, Lamuela-Ravents, Viladomat, Bastida, Codina. RapidCommunications in MassSpectrometry, 2004, 18, 553-563
Artichoke waste
Apple pomace
Tagetesmaxima
Torras-Claveria, Juregui, Bastida, Codina, Viladomat. Journalof AgriculturalandFoodChemistry, 2007, 55, 8436-8443
Lavandin
Metabolomics
Luna, Pigni, Torras-Claveria, Monferran, Maestri, Wunderlin, Ferensin, Bastida, Tapia. Journalof FoodCompositionandAnalysis, 2013, 31, 120-128
Polyphenols LC-MS/MS and LC high resolution MS2000 - today
Torras-Claveria, Juregui, Codina, Tiburcio, Bastida, Viladomat. Plant Science, 2012, 182, 71-78
Gasca, Cabezas, Torras, Bastida, Codina. FreeRadicals andAntioxidants, 2013, 3, S55-S61
Tobacco
Pavoniasepioides
Ramorinoagirolae
De Andrade, Berkov, Viladomat, Codina, Zuanazzi, Bastida. Molecules, 2011, 16, 7097-7104
Hippeastrumpapilio
De Andrade, Pigni, Torras-Claveria, Guo, Berkov, Reyes-Chilpa, El Amrani, Zuanazzi, Codina, Viladomat, Bastida. Revista Latinoamericanade Qumica, 2012, 40, 83-98
GenusHippeastrum
De Andrade, Guo, Font-Bardia, Clavet, Dutilh, Viladomat, Codina, Nair, Zuanazzi, Bastida. Phytochemistry, 2014, 56, 188-195
Hippeastrumaulicumand calyptratum
Guo, de Andrade, Pigni, Torras-Claveria, Tallini, Borges, Viladomat, Nair, Zuanazzi, Bastida. HelveticaChimicaActa, 2016, 99, 143-147
Sebben, Giordani, de Andrade, Berkov, Osorio, Sobral, de Almeida, Henriques, Bastida, Zuanazzi. Revista Brasileirade Farmacognosia, 2015, 25, 353-355
Hippeastrumbreviflorum
Reyes-Chilpa, Berkov, Hernndez-Ortega, Jankowski, Arseneau, Clotet-Codina, Est, Codina, Viladomat, Bastida. Molecules, 2011, 16, 9250-9533
Zephyranthesconcolor
Osorio, Berkov, Brun, Guo, Codina, Viladomat, Cabezas, Bastida. PhytochemistryLetters, 2010, 3, 161-163
Phaedranassadubia
De Andrade, Giordani, Torras-Claveria, Pigni, Berkov, Font-Bardia, Calvet, Konrath, Bueno, Sachett, Dutilh, Borges, Viladomat, Henriques, Nair, Zuanazzin, BastidaPhytochemistryReviews, 2016, 15, 147-160
Brazilian Amaryllidaceaeas a source of acetylcholinesterase inhibitory alakaloids
Guo, Pigni, Zheng, de Andrade, Torras-Claveria, Borges, Viladomat, Codina, BastidaNatural ProductCommunication, 2014, 9, 1081-1086
Bioactive alkaloids in Lycorisspecies
Pharmacological activity
Increase of interest in pharmacological activity of Amaryllidaceaealkaloids since GAL approval by FDA
Anti-tumoralAnti-parasiticAcetylcholinesterase inhibitionDocking studies
Pharmacological activity
Some Amaryllidaceaehave been historically used in symptomatologiesidentified as cancer
Citotoxicitytest in anti-tumor compounds
Toxicity test in ArtemiasalinaPrevious evaluation of AmaryllidaceaecompoundsReproducible and comparable values
Pharmacological activity
Citotoxicitytest in anti-tumor compounds
Evaluation in 60 cell lines
Pharmacological activity
Citotoxicitytest in anti-tumor compounds
Pharmacological activity
Apoptosis Selectivity
NH
O
O
OH
OH
HO
OH
OH O
Pancratistatin
McLachlan, Kekre, McNulty, Pandey. Apoptosis2005, 10, 619-630Kekre, Griffin, McNulty, Pandey. CancerChemother. & Pharmacol. 2005, 56, 29-38
Selective apoptosis of tumoral cellsMinimal effects in normal cellsEarly activation of Caspasa-3Fast apoptosis in Human Linfomacells
Pharmacological activity
Apoptosis SAR studies Selectivity Hepatoma (5123tc)
N
O
O
OH
HO
HN
O
O
OAc
HO
HN
O
O
H
OH
N
O
O
H
OMe
OMe
HO
N
O
O
H
OH
OMe
N
O
O
H
OH
OMe
N
O
O
HN
O
O
H
HO
OMeO
OMe MeO
OOMe
OH
amabiline(3)
josephinine(4)
crinine(5)
ambelline(6)
crinamine(7)
haemanthamine(8)
epoxyambelline(9)
6 -hydroxyundulatine(10)
25mM 48h treatment
0
10
20
30
40
50
60
70
80
90
100
C A1 A2 A3 A4 A5 A7 A8 A10
% A
po
pto
tic
c 3 4 5 7 8 6 9 10
C 3 4 5 7 8 6 9 10
McNulty, Nair, Codina, Bastida, Pandey, Gerasimoff, Griffin. Phytochemistry2007, 68, 1068-1074
% A
po
pto
sis
CRININE-HAEMANTHAMINE
Pharmacological activity
Apoptosis SAR studies Selectivity Hepatoma (5123tc)
McNulty, Nair, Codina, Bastida, Pandey, Gerasimoff, Griffin. Phytochemistry2007, 68, 1068-1074
Pharmacophoricrequirements
Presence of ethanobridge -5,10bMethoxylgroup tolerance in or in C-3Hydroxyl group in position 11
N
OHO
O
OMe311
H
Pharmacological activity
Anti-parasitic
Parasite Disease Infectectedpeople Detection
Plasmodiumfalciparum
Malaria 300 millions (2 millions death/year)
HypoxanthineH3 incorporation
Leishmaniasp. Leishmaniasis 12 millions Microscopy
Trypanosoma brucei-rhodesiense
Sleeping sickness 300.000(100%mortality)
Alamarblue assay (colorimetry)
Trypanosoma cruzi Chagas disease 18 milions Colorimetry
Spencer et al., Lloydia1947, 10, 145-174
Pharmacological activityAnti-parasitic
Parasite: strain: stage: reference drug:
T. b. rhodesiense STIB 900 trypomastigotes Melarsoprol
T. cruzi Tulahuen C4 amastigotes Benznidazole
L. donovani MHOM-ET-67/L82 amastigotes Miltefosine
P. falciparum K1 IEF Chloroquine
Cytotoxicity L6 Podophyllotoxin All values as: g / mL
ID No IC-50 ref drug IC-50 ref drug IC-50 ref drug IC-50 ref drug IC-50 ref drug
Ungeremine 0,979 0,003 0,85 0,22 >30 0,17 0,0933 0,0321 17,3 0,008
Buphanamine 28,75 0,003 >30 0,22 >30 0,17 4,31 0,046 >90 0,008
Zefbetaine 39,22 0,003 >30 0,22 >30 0,17 >5 0,046 >90 0,008
Galanthamine 37,92 0,003 >30 0,22 >30 0,17 >5 0,046 >90 0,008
Pdub Extract 27,43 0,003 >30 0,22 >30 0,17 >5 0,046 >90 0,008
P. falc. K1 Cytotox. L6T.b.rhod. T. cruzi L.don. axen.
N
R1O
R2O
O
OMeO
NMeN
HO
O
O
OMe
H
OH
Buphanamine GalanthamineUngeremine: R1+R2=CH2Zefbetaine: R1=R2=Me
Osorio, Robledo, Bastida. TheAlkaloids2008, 66, 113-190
Pharmacological activity
Strategy to palliate Alzheimer symptomatology
Acetylcholinesterase activity inhibition
Drugs with cholinergic activity
H3CO
NCH3
O
OH
Ellmanin vitro assay to test Amaryllidaceaealkaloids and plant extracts
Galanthamine