Two examples of bio-guided isolation, synthesis and ...

Two Examples of
Bioguided Isolation, Synthesis and Pharmacomodulation of
Natural Products from Annonaceae and Rutaceae
Dr Bruno FIGADERE
Laboratories of Natural Substances Chemistry
(Pharmacognosy), UMR 8076-BioCIS
Faculty of Pharmacy, University Paris-Sud
Thai-Franco Seminar on Natural Substances
27 September 2005
BioCIS (UMR CNRS 8076)

Laboratory of Chemistry of Natural Substances
- Extraction of natural products with biological properties
- Natural products and analogues: synthesis and semi-synthesis
- Total Synthesis of Natural Products
- Antiparasitic Chemiotherapy
BioCIS (UMR CNRS 8076)

Bioguided Extraction - Leishmanicidal Compounds:
The Chimanines, 1st Example
- in Bolivia, Galipea longiflora (Rutaceae) is traditionally used
against cutaneous leishmaniasis
- bioguided fractionation and purification led to biological active
alcaloides
N
N
OMe
O
N
N
N
N
O
O
(Fournet, A.et al J. Nat. Prod. 1993, 56, 1547-1552)
BioCIS (UMR CNRS 8076)

Synthesis of 2-substituted quinolines (part 1)
H 2 O 2
ClCOOiBu, t he n RMgBr
N
N
O
35-90 %
(Fakhfakh,M.A.. et al Tetrahedron Lett. 2001, 42, 3847)
N
R
N
(67%)
N
(79%)
N
(73%)
N
(53%)
N
(82%)
N
(68%)
N
(63%)
N
(35%)
N
(75%)
SiMe 3
N
(66%)
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Synthesis of 2-substituted quinolines (part 2)
Wittig
N R N
O
Br
N
(89%)
Br
N
(80%)
CO 2 Et
N
CHO
N
OMe
(64%)
(80%)
N C 5 H 11
N CO 2 Et
(84%) (63%)
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N CN N NO 2
(80%) (70%)
(Fakhfakh,M.A. et al Synth. Commun. 2002, 32, 2863)

Synthesis of 2- or 3-substituted quinolines (part 3)
N
Cl
RMgX, Fe(acac) 3
THF, -10 °C-0 °C
N
R
N
76%
N
36%
OH
75% N
N
75%
N
70%
(Seck M. et al Tet. Lett. 2004, 45, 1881-1884)
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Synthesis of 2- substituted quinolines (part 4)
N
OH
Z
F
NO 2
N(CH 3 ) 2
OCH 3
N
H
N
Z
N
O
N
NCH 3
N
OH
CO 2 Me
N
O
Bu
N
O
OEt
N
OH
OMe
OMe
R
N
R
H
Ph
CO 2 Me
N
Z
F
NO 2
N(CH 3 ) 2
OCH 3
N
R
N
N
O
O
n
O
n=1or 7
O
N
N
N
N
O
O
N
N
N
N
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In vitro activity of synthetic quinolines (IC 50 µM)
L. am az onensis
L. infant um L. donovani
cytotoxicity (macrophages
N
5
32 -
41
N
CHO
4
2 -
9
N
OH
4
2 -
34
N
Br
3
2 -
13
21
32 1,2
40
N
N
CO 2 Me
59
26 3,3
>200
N
37
12 -
74
amphotericine B 0,12
Gluca nt im e 20
pentamidine -
BioCIS (UMR CNRS 8076)
- -
7 -
- 1
-
-
-

In vivo activity of synthetic quinolines
L. infantum Balb/c infected mice (10 days treatment by oral administration 2x12.5
mg/kg/d)
Gluca nt im e
% of reduction of parasites
in the liver
- 65,3
% of reduction of parasites
in t he s ple e n
+ 6,0%
N
- 1 3,3
+ 95,6
N SiMe 3
- 3 7,8
- 23,8
N
- 60,0
- 47,6
N
OMe
- 6 0,0
- 64,3
N
CHO
- 91,6
- 47,6
N
CH 2 OH
- 51,1
- 3 8,1
N Br
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- 24,4 - 33,3
(Fakhfakh, M.A. e t al PCT/FR02/00140 ; PCT Int. Appl. WO 02 57,238 ; CA 2002, 137, 125093m)

Bioguided Extraction - Cytotoxic Compounds:
The Acetogenins of Annonaceae, 2nd Example
O OH OH
O
1
n
O
S
O
O
O
O
O
O
type A1
O
OH
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goniocin (type D1)
OH
reticulatamol (type E)
R, R, R, R : threo-trans-threo
S,R,S,R : threo-cis-threo
n=1, m=3, threo-trans-threo : trans-solamin
n=1, m=3, threo-cis-threo : cis-solamin
n=3, m=3, threo-trans-threo : reticulatacin
n=3, m=3, threo-trans-erythro : reticulatain 1
n=5, m=1, threo-trans-erythro : reticulatain 2
n=5, m=1, threo-trans-threo : uvariamicin 3
OH
OH
O O
O
isomolvizarin (type B2)
OH
O O
Figadère, Cortes et al. Nat. Prod. Rep. 2005, 22, 269-303
m

Acetogenins of Annonaceae
O
O
O
OH
O O OH
O
O
O
rolliniastatin 2
OH
O
OH
iso-rolliniastatin 2
antitumoral activity on grafted mice
OH
Products
rolliniastatin 2
iso-rolliniastatin 2
paclitaxel
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Quantity(/day)
µg/kg
200
100
50
25
12
800
400
200
100
50
25
25000
17500
15000
7000
4400
% T/C Variation of weight
TOX
106
138
125
113
144
144
125
113
113
100
90
105
139
114
117
(g/mouse)
-2,9
-2,6
-0,4
0,6
0,9
0,5
1,1
1,8
1,6
2,1
1,7
-4,4
-2,4
-2,5
-1,1
-0,9

Semisynthesis- Perspectives and collaborations
Acetogenins of’Annonaceae: collaboration with Pasteur Institute, ICSN (Gif-s-Yvette, France)
fluorescent probe (visualisation)
Proteomic
N 3
OR
O
O
OR
O
O
protection, azide formation
OH
OH
O
O
OH
O
O
squamocine
OH
OH
O
KMnO 4
O
OH
OH
R
radical decarboxylation +
O
R
O
R R
OH OH
OH O O
O O
R
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R
O
R

Biomimetic Total Synthesis of Solamin (part 1)
Z
E
O
O
pseudo- muricadienin
AD-mix β
HO
O
O
OH
oxidation
O
O
O
O
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OH
15 R
16 R
O
trans-solamin
or
OH
20 R
19 R
OH OH
15 S
O 20 R
16 S 19 R
cis-solamin
or
diastereomers

Biomimetic Total Synthesis of Solamin (part 2)
MeO
O
Z
E
MeO
O
4 steps
Z
Br
+
Metal
E
2 steps
O
O
OH
OH
Figadère, Tetrahedron Lett. 1999, 40, 4041 and Let t . Org. Chem . 2004, 1 ,
316-322.
O
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Conclusion
- From several plants,bio-guided isolation led to natural 2-substituted quinolines that have shown
promising in vivo activity against a murine model of visceral leishmaniasis; bioguided synthesis
then allowed us to obtain suitable products for preclinical trials
- From several Annonaceae, bio-guided isolation has afforded a series of new cytotoxic compounds
the acetogenins of Annonaceae; semi-synthesis, and total synthesis gave so far, some information
on the structure-activity relationship, as well as on the cellular target.
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BioCIS (UMR CNRS 8076)

BioCIS (UMR CNRS 8076)

BioCIS (UMR CNRS 8076)