51
1 Group Meeting Montreal, February 22, 2012 Sonia Diab
1
Group Meeting
Montreal, February 22, 2012
Sonia Diab
2CONSTANTINE
3
CAEN (Basse-Normandie
4
ROUEN (Haute-Normandie)
5
SYNTHESIS OF DIFLUOROPHOSPHONATES BY
CARBANIONIC AND RADICAL PROCESS FOR THE
PREPARATION OF THYMIDINE PHOSPHORYLASE
INHIBITORS
MMM
PhD Supervisor: Pr. Thierry. LEQUEUXLaboratoire de Chimie Moléculaire et Thio-organique. UMR-CNRS 6507
6 Boulevard du Maréchal Juin 14050 CAEN, France
Sonia DIABMontreal, February 22, 2012
6
IntroductionIntroduction
o Tumoral AngiogenesisTumoral Angiogenesis
PD-ECGF: : Platelet-derived endothelial cell growth factor
• Protein involved in angiogenesis of cancerous cells.• Identified as Human Thymidine Phosphorylase (HTP)
o Propperties of Human Thymidine PhosphorylasePropperties of Human Thymidine Phosphorylase
(a) Folkman J and al. J Biol Chem 1992 ; 267 : 10931-10934, (b) Ishikawa, F. and al. Nature 1989, 338, 557-562.
O
OH OH
N
PO
OH
HO
HO
12 Å
HN
O
O
domain
adomain
Opened Conformation
O
OH OH
N
PO
OH
HOHO
4 Å
HN
O
O
domain
adomainClosed
Conformation
(a) Walter, M. R. And al. J. Biol. Chem. 1990, 265, 14016-14022. (b) Norman, R. A. And al. Structure 2004, 12, 75-84.
Inhibitor of TP
Base
Spacer 4-12 Å
X POH
OH
O
7
o Multisubstrats InhibitorsMultisubstrats Inhibitors
Synthesis of new fluorinated inhibitors of TP
Inhibitors of Thymidine phosphorylaseInhibitors of Thymidine phosphorylase
• Very few TP inhibitors described
• Phosphonate as mimics of the phosphate
function
N
NH
H2N
O
O
Br
PO3H2
TP64Ki 35 M
N
NHO
OH
N
PO3H2
TP65Ki 20 M
HN
N
O
O
O
R
PO3H2
PMEA derivativeR = CH3, CH2OH
Ki 2- 3 M
NH
N
O
OO
OO
HO
H
PO3H2
GanemKi 0.24 M
• No fluorinates TP inhibitors described to date
Blazarini, J. and al. J. Med. Chem. 2000, 43, 971-984.
o Analogie phosphate / phosphonate / fluorophosphonateAnalogie phosphate / phosphonate / fluorophosphonate
• Difluorophosphonates best isopolar and isosteric phosphate analogue
pKa2 = 6.45 pKa2 = 7.65 pKa2 = 5.65
PO
HO
OO
PO
HO
CH2O
PO
HO
CF2O
Blackburn, M. G. ; England, D. A. ; Kolkmann, F. J. Chem. Soc. Chem. Comm. 1981, 930-392
8
Choice of structures to synthesizeChoice of structures to synthesize
Spacer
T PH I nhi bi t or s
P
O
HO
HO
F F
Base
4- 12 Å
OP
O
O
N
HN
O
O
Variable distances
O
O
HO
HO
OP
O
O
N
HN
O
O
O
HN
N
O
O
CF2P(O)(OH)2
( )n
Aliphatic nonfunctionalized
chain
Variable distances
OP
O
O
O
N
HN
O
O
HO
O
O
N
HN
O
O
HO
CF2P(O)(OH)2
( )n
Variable distances
Aliphatic functionalized chain
OP
O
O
N
HN
O
O
O
O
N
HN
O
O
CF2P(O)(OH)2( )n
HO
HO
O
N
HN
O
O
HO
HO
CF2P(O)(OH)2( )n
O
HO
HO Variable distances
Mimic the transition state
OP
O
O
N
HN
O
O
O
N
HN
O
O
CF2P(O)(OH)2( )n
N
HN
O
O
( )n
NN
N
CF2P(O)(OH)2
arom Variable distances
Presence of an aromatic
9
OP
O
O
N
HN
O
O
Variable distances
O
HN
N
O
O
CF2P(O)(OH)2
( )n
O
N
HN
O
O
HO
CF2P(O)(OH)2
( )n
N
HN
O
O
CF2P(O)(OH)2( )n
N
HN
O
O
( )n
NN
N
CF2P(O)(OH)2
O
N
HN
O
O
CF2P(O)(OH)2( )n
HO
HO
O
N
HN
O
O
HO
HO
CF2P(O)(OH)2
( )n
Aliphatic nonfunctionalized
chain
Aliphatic functionalized chain
Presence of an aromatic
Mimic the transition state
O
HO
HO
Spacer
T PH I nhi bi t or s
P
O
HO
HO
F F
Base
4- 12 Å
Choice of structures to synthesizeChoice of structures to synthesize
10
Les Families of TP Inhibitors Les Families of TP Inhibitors
o Family I :Family I : Linear Inhibitors Linear Inhibitors
o Family II :Family II : Conformationally constrained Inhibitors Conformationally constrained Inhibitors
How to introduce the difluoromethylphosphonate function
P
ORO
ROF F
4- 12 Å
Base( )n
R
P
ORO
ROF F
OH( )n
P
ORO
ROF F
I onic routeR
P
ORO
ROF F
4- 12 Å
Base( )n
R
P
ORO
ROI
F F
Base( )n+ P
ORO
ROF F
Radical route
P
ORO
ROF F
4- 12 Å
Base( )n
R
P
ORO
ROF F
OH( )n
P
ORO
ROF F
I onic routeR
P
ORO
ROI
F F
Base( )n+ P
ORO
ROF F
Radical route
P
ORO
ROF F
4- 12 Å
Base( )n
R
Base
( )nP
OH
OOH
FF( )n
P
ORO
ROF F
I onic route
HO
( )nP
OR
OOR
FF( )n
P
ORO
ROI
F F+ P
ORO
ROF F
Radical route
11
General methods for the synthesis of fluorophosphonatesGeneral methods for the synthesis of fluorophosphonates
(a) Burton, D. J. et al. Chem. Rev. 1996, 96, 1641-1715. (b) Matulic-Adamic, J. et al. J. Org. Chem. 1995, 60, 2563-2569.
(a) Li, A.-R. ; Chen, Q.-Y. Synthesis 1996, 606-608. (b) Yokomatsu, T. et al. J. Org. Chem. 1996, 61, 7207-7211.
Herpin, T. F. Et al. Chem. Commun. 1996, 613-614.
(a) Solas, D. et al. J. Org. Chem. 1996, 61, 1537-1539. (b) Lequeux, T. et al. Org. Lett. 2001, 3, 185-188
(a) Blades, K. et al. Chem. Commun. 1996, 1457-1458. (b) Xu, Y. et al. Org. Lett. 2002, 4, 4021-4024.
CF2P
O
RO
RO CF2CF2
P
O
RO
RO
ZP
O
RO
RO
Z = C(O), CH2
PCF2
ORORO
Y
Y = H, M, X, S, Se
Direct nucleophilic or radical introduction
Construction from
gem- difluoroalkenes
Nucleophilic or electrophilic fluorination
Using phosphonofluorinated
"building blocks"
PCF2
ORORO
PCF2
ORORO
P
12
Limits: based on the use of HCFC or CFCLimits: based on the use of HCFC or CFC
• Halons and freons: regulated and prohibited for use since the Kyoto and Montreal protocole
Need to find other methods that avoid the use of ecotoxic materials.
(a) McCulloch, A. J. Fluorine Chem. 1999, 100, 163-173. (b) Obayashi, M. ; Ito, E. ; Kondo, K. Tetrahedron Lett. 1982, 23, 2323-2326. (c) Burton, D. J. et al. Chem. Lett. 1982, 755-758. (d) Burton, D. J. ; Takei, R. ; Seiji, S. J. Fluorine Chem. 1981, 18, 197-202.
CF2P
O
RO
RO CF2
PCF2
ORORO
MXnOrganometallic
reagents
DifluoroalkenesTransition
metals
HCF2Cl or CF2Br2
CF2Br2PCF2
ORORO
MCF2I2CF2Br2
13
Previous work of the Previous work of the laboratorylaboratory
o New method to generate the anionNew method to generate the anion
P
OO
OF F
LiP
OO
OS
3HF-NEt3, ZnBr2,
CH3CN, ref lux
P
OO
OS
62%
F F
P
OO
OS
Cl Cl
SO2Cl2,
CH2Cl2
85%
t-BuLi,
-78°C
Henry-dit-Quesnel, A. ; Toupet, L. ; Pommelet, J. C. ; Lequeux, T. Org. Biomol. Chem. 2003, 1, 2486-2491
Ozouf, P. ; Binot, G. ; Pommelet, J. C. ; Lequeux, T. Org Lett. 2004, 6, 21, 37-47.
o Study of th reactivity of the anion depending on its source Study of th reactivity of the anion depending on its source
• E+ : TMSCl, aldehyde, ketone, CS2 The same reactivity
• E+ : , , , DMF Different reactivity
P
ORO
ROF F
E P
ORO
ROF F
E
OR O
O
• Fluorosulfide: Novel source of the anion developed in Lequeux’s Laboratory
P
OO
OH
F F
1/ LDA, -78°C
2/
BF3-Et2O
O
Compound 1
No reaction P
OO
OS
F F
1/ t -BuLi, -78°C
2/
BF3-Et2O
O
Compound 2
Opening of
Oxacycles
Anion can open oxacycles to prepare corresponding alcohols.
14
Family I:Family I:
Inhibitors with a linear chainInhibitors with a linear chain
P
ORO
ROF F
OH( )n
P
ORO
ROF F
I onic routeR
P
ORO
ROI
F F
Base( )n+ P
ORO
ROF F
Radical route
P
ORO
ROF F
4- 12 Å
Base( )n
R
15
Hydroxyphosphonates: Key molecules for the synthesis of our inhibitorsHydroxyphosphonates: Key molecules for the synthesis of our inhibitors
• Formation of hydroxydifluoromethylphosphonates with various size
• Opening reations of oxacycles
4- 12 Å
P
OO
O OHF F
P
OHO
HO BaseF F
( )n( )n
n = 1
P
OO
OF F
OH
OH
Previous laboratory work
n = 2
OR
1,2- cyclic sulfate
R1 R2
OS
O
O O
Epoxide
n = 3
OR'
R
Oxetanen = 4
O
THF
n > 4
O O
( )n
Lactone
16
o 1,2-cyclic sulfates opening1,2-cyclic sulfates opening
Preparation of Preparation of -hydroxydifluoromethylphosphonates-hydroxydifluoromethylphosphonates
P
OHO
HOF F
4- 12 Å
Base
P
OO
OF F
HN
69%
Diab, S. A. ; Sene, A. ; Pfund, E. ; Lequeux, T. Org. Lett. 2008, 10, 3895-3898.
( )4
P
OO
OF F
Cl
OH
P
OO
OF F OH
53%
79% 83%
76%
P
OO
OF F OH
P
OO
OF F
OBn
OH
P
OO
OF F
O
OH
OMe
69%
71%
P
OO
OF F OH
2/ H2SO4
OS
O
O O
RP
OO
OLi
F F
1/ -78°C, THF, 15 min.
P
OO
OF F
R
OS
O
O
O
P
OO
OF F
R
OH
53- 83%
(1.3 eq.)
17
o Oxetanes openingOxetanes opening
Preparation of Preparation of -hydroxydifluoromethylphosphonates-hydroxydifluoromethylphosphonates
P
OHO
HOF F
4- 12 Å
BaseO
P
OO
OF F
OH
63%
P
OO
OLi
F F
1/
Et2O, -78°C BF3-Et2O, 5 min.2/ NH4Cl
O
OBnO
P
OO
OF F
OBn
57%
P
OO
OF F
OBn
65%OH
OH
BnO1/ t-BuLi, -78°C, Et2O
5 min.2/ BF3-Et2O3/ NH4Cl
OO
OO
OOH
OO
50%
P
FF
O
O
O
P
OO
OLi
F F
Diab, S. A. ; Sene, A. ; Pfund, E. ; Lequeux, T. Org. Lett. 2008, 10, 3895-3898.
Ozouf, P. ; Binot, G. ; Pommelet, J. C. ; Lequeux, T. Org Lett. 2004, 6, 21, 37-47.
18
• No reaction
• The size of oxacycles opening is limited to the THF
o THF openingTHF opening
Exploit other alternatives to increase the size of the spacer
Preparation of Preparation of -hydroxydifluoromethylphosphonates-hydroxydifluoromethylphosphonates
o THP openingTHP opening
P
OHO
HOF F
4- 12 Å
Base
P
OHO
HOF F
4- 12 Å
Base
P
OO
OLi
F F
79%
O
1/ Complexe:
BF3-
30 min, -78°C2/ NH4Cl
P
OO
OF F
OH
Ozouf, P. ; Binot, G. ; Pommelet, J. C. ; Lequeux, T. Org Lett. 2004, 6, 21, 37-47.
P
OO
OLi
F F
1/ Complexe:
BF3-
-78°C2/ NH4Cl
ONo reaction
19
o Lactones opening: Lactones opening: -caprolactone and -caprolactone and -lactone-lactone
• Lactones opening leads to the formation of hydroxyketodifluoromethylphosphonates
Preparation of Preparation of -hydroxydifluoromethylphosphonates-hydroxydifluoromethylphosphonates
P
OHO
HOF F
4- 12 Å
Base( )n
P
OO
OLi
F F
ratio: 85 / 15
n = 1, 258- 61%
1/ Et2O, -78 °C
BF3-Et2O (2,0 eq.) +
(1,3 eq.) 10min2/ NH4Cl -78 °C -> r.t.., 1h
+P
OO
OF F
O
( )n
( )nOH
OOH
PO
OO
F F
( )n
O O
P
OO
OLi
F F-caprolactone
P
OO
OF F
O
O
-lactone
OO
O
OH
6 C
( )3 P
OO
OF F
O
OH
7 C
( )4
Introduction of the nucleic base
20
Preparation of fluorinated analogues of linear acyclonucleotidesPreparation of fluorinated analogues of linear acyclonucleotides
o Approach 1: Mitsunobu ReactionApproach 1: Mitsunobu Reaction
• Direct introduction of the nucleic base by Mitsunobu reaction Moderate yields and nonreproducible reaction
Using TMG to introduce the nucleic base
o Approach 2: Activation by tosylateApproach 2: Activation by tosylate
Conditions tested:
1/ NaH/DMF/reflux Low yields (< 35%)
2/ K2CO3/DMF/r.t. No reaction
3/ TMG/DMSO/r.t. Best results
Diab, S. A. ; Sene, A. ; Pfund, E. ; Lequeux, T. Org. Lett. 2008, 10, 3895-3898.
56- 78%
NH
N(Me)2(Me)2N
DMSO, 16-24h, r.t.
1/
2/ CH3NH2, MeOH, 18h, r.t.
P
OO
OF F
( )n
OTsP
OO
OF F
( )n
N
NH
O
O O
N NH
O
OP
OO
OF F
( )n
OH
TsCl, NEt3,
CH2Cl2, r.t., 24h
70- 89%
21
Preparation of fluorinated analogues of linear acyclonucleotidesPreparation of fluorinated analogues of linear acyclonucleotides
NH
NH
O
O
NH
NH
O
O
NH
NH
O
O
Br
N
NN
NH
Cl
NH2N
NN
NH
Cl
(i-PrO)2P
F F
O
N
NH
O
O
O
F FNH
N O
O
(i-PrO)2P
O
( )n
n = 1, 71%n = 2, 67%
(i-PrO)2P
O
F Fn = 1, 72%
n = 2, 67%
( )n
N
NN
N
Cl
(i-PrO)2P
O
F Fn = 1, 60%
n = 2, 68%
( )n
N
NN
N
Cl
NH2
F FNH
N O
O
(i-PrO)2P
O
( )n
n = 2, 78%
F FNH
N O
O
(i-PrO)2P
O
( )n
n = 2, 75% Br
( )n
n = 1, 56%n = 2, 62%
Diab, S. A. ; Sene, A. ; Pfund, E. ; Lequeux, T. Org. Lett. 2008, 10, 3895-3898
o Introduction of the nucleic base Introduction of the nucleic base
22
• Late introduction and at the same time of the difluoromethylphosphonate group and the nucleobase
Access to long-chain inhibitorsAccess to long-chain inhibitors
P
OHO
HOF F
4- 12 Å
Base( )n
o Synthesis according to the first strategy: ionic process Synthesis according to the first strategy: ionic process
( )n
• Early introduction of the difluoromethylphosphonate group
o Synthesis according to the second strategy: radical processSynthesis according to the second strategy: radical process
( )n
• Late introduction of the nucleobase
P
OHO
HOF F
( )nP
OHO
HOF F
Base( )n
P
OHO
HOF F
Base( )n
P
OHO
HOF F
4- 12 Å
Base( )n
P
ORO
ROI
F F
Base( )n+ P
ORO
ROF F
Radical route
23
o Preparation of iododifluoromethylphosphonatePreparation of iododifluoromethylphosphonate
• Three initiator systems were used
• Regioselective reaction
Radical reactionRadical reaction
• Chalcogen-halogen exchange avoids the use of HCFCs
o Reaction with Reaction with nn-octene-octene
Functionalized alkenes
Henry-dit-Quesnel, A. ; Toupet, L. ; Pommelet, J. C. ; Lequeux, T. Org. Biomol. Chem. 2003, 1, 2486-2491
Sène, A. ; Diab, S. ; Hienzsch, A. ; Cahard, D. ; Lequeux, T. Synlett. 2009, 981-985.
1/ tBuLi, THF, -78 °C
2/ I 2, 1h, -78 °C
81%
P
OO
OS
F F
P
OO
OI
F F
Methodea (A) or (B) or (C)P
OO
OI
F F
C6H13
P
OO
OF F
C6H13
I
a Methodes:
(A) Na2S2O4, NaHCO3, CH3CN/ H2O (5/ 3), rt., 18h.
(B) BEt3 (0,3 eq., 1M in n-hexane), CH2Cl2, r.t., 1h.
(C) Lauroyl peroxide (0,3 eq.), C2H4Cl2, 80 °C, 3h.
(A), 75%(B), 76%(C), 45%
24
o Addition on the allylthymineAddition on the allylthymine
Addition of the radical phosphonodifluoromethyle on the alkenesAddition of the radical phosphonodifluoromethyle on the alkenes
Lauroyl peroxide (0,3 eq.), C2H4Cl2, 80 °C, 3h
P
OO
OI
F F
P
OO
OF F I
71%
N NH
O
ON
HNO O
o Addition on functionalized alkenes with long-chainAddition on functionalized alkenes with long-chain
P
OO
OI
F FNa2S2O4 (4 eq.), NaHCO3, CH3CN/ H2O (5/ 3), r.t., 24h.
Base
P
OO
OF F
BaseI
P
OO
OF F I
P
OO
OF F I
73%
NH
N O
O 76%
N
N
N
N
Cl
NH2
• Reaction performed in the presence of purine and pyrimidine base
25
Family II:Family II:
Conformationally constrained InhibitorsConformationally constrained Inhibitors
Base
( )n
P
ORO
ROF F
I onic route
POH
OOH
FF( )n
HO
( )nP
OR
OOR
FF( )n
P
ORO
ROI
F F+ P
ORO
ROF F
Radical route
26
Retrosynthsis of the structures to be synthetizedRetrosynthsis of the structures to be synthetized
• Mimic the approach of the phosphate group
• Conformation frozen by a benzene ring
• Conformation frozen by a triazole moiety
OBase
P
F FOH
OOH
O OH
P
F FOR
OOR
O
P
ORO
ROF F
+
Radical route
O
OS
O
O
Base
P
O
OH
OHFF
P
ORO
ROF F
I onic route
OH
P
O
OR
ORFF
+
Base
P
ORO
ROF F
I onic route
( )n
NN
N
( )n
Base
( )n +
P
FF
O OHOH
( )n
N3
P
FF
O OROR
( )n
OH
P
FF
O OROR
27
Conformation frozen by a furan ringConformation frozen by a furan ring
• Differents initiators were used The product was only observed in the presence of sodium dithionite
• The trans isomer is predominant (confirmed by NMR experiments HOESY 1H{19F})
• Heterocycle to block the conformation
o Radical addition on 2,3-dihydrofuranRadical addition on 2,3-dihydrofuran
P
OO
OI
F F
89%
Na2S2O4 (4 eq.), NaHCO3, CH3CN/ H2O (5/ 3), r.t., 24h.
OO OH
P
F FO
OO
OI CF2 P
CF2 P
CF2 P
O
CF2 P
I
- I
O
CF2 P
P = P(O)(Oi Pr)2
H2O O
CF2 P
OH
t r ans/ ci s 93/ 7
28
• Diastereoselective reaction (trans configuration)
• Formation of two regioisomers N 1 and N 3
o Introduction of the nucleic baseIntroduction of the nucleic base
Conformation frozen by a furan ringConformation frozen by a furan ring
O OH
P
F FO
OO
Ac2O, NEt3,
DMAP, CH2Cl2, r.t., 3h
89%
O O
P
F FO
OO
O
N
N
O
O
Si
Si
SnCl4, -15°C, 2h
O O
P
F FO
OO
O
ON
P
F FO
OO
HN
O
O+ O
N
P
F FO
OO
HN
O O
I somer N1 + I somer N3 = 76%
O
OS
O
O
P
OO
OS
F F
1/ t -BuLi, THF, -78 °C
2/
15 min3/ H2SO4 20%
52%
OHP
O
O
O
F F
72%
NP
O
O
O
F F
HN
O
O
1/ TsCl, NEt3, CH2Cl2, r.t., 24h
2/ protected base, TMG, DMSO, r.t., 16h3/ CH3NH2, MeOH, r.t., 18h
29
Conformation frozen by a benzene ring Conformation frozen by a benzene ring
o Preparation of 1,4-cyclic sulfatePreparation of 1,4-cyclic sulfate
o 1,4-cyclic sulfate opening1,4-cyclic sulfate opening
• First exemple of 1,4-cyclic sulfate opening by difluoromethylphosphonate anion
(a) Soai, K. ; Ookawa, A. J. Org. Chem. 1986, 51, 4000-4005. (b) Gao, Y. ; Sharpless, K. B. J. Am. Chem. Soc. 1988, 110, 7538-7539.
1/ LiBH4, MeOH,
Et2O, 45 °C, 30 min.2/ HCl 1M 81%
O
O
O
O
OH
OH
1/ SOCl2, CH2Cl2, NEt3, 0 °C, 30 min.
2/ NaI O4, RuCl3-H2O (0,02%), H2O, CH3CN, r.t., 1h 85%
O
OS
O
O
30
Conformation frozen by a triazole ringConformation frozen by a triazole ring
o 1,3-dipolar cycloaddition1,3-dipolar cycloaddition
N
71%
P
F F
OOO N
N
NHN
O
O
N
79%
P
F F
OOO N
N
N
N
N
N
Cl
NH2F F
P
OOO
87%
NN
NNHN
O
O
F F
P
OOO
74%
NN
NN
N
N
N
Cl
NH2
N
96%
P
F F
OOO N
N
NHN
O
O
N
75%
P
F F
OOO N
N
N
N
N
N
Cl
N
71%
P
F F
OOO N
N
N
N
N
N
Cl
NH2F F
P
OOO
93%
NN
NNHN
O
OF F
P
OOO
85%
NN
NNHN
O
O
F F
P
OOO
82%
NN
NN
N
N
N
Cl
F F
P
OOO
75%
NN
NN
N
N
N
Cl
NH2
Diab, S. A. ; Hienszch, A. ; Lebargy, C. ; Pfund, E. ; Lequeux, T. Organic and Biomolecular Chemistry, 2009, 21, 4481-4490.
P
OO
OF F
N3( )nP
OO
OF F
OTs( )n
n = 2, 3, 478- 85%
NaN3, DMF,
r.t., 16h
n = 2, 3, 480- 90%
Base+
CuSO4
(5 mol%)sodium ascorbate
(10 mol%)
t BuOH/ H2O 1/ 1,
r.t., 12-72 h
P
OO
OF F
N( )n
NN
Base
71- 96%
31
Tests of inhibition of Tests of inhibition of
Thymidine phosphorylase from Thymidine phosphorylase from E.ColiE.Coli
32
In vitroIn vitro inhibition of TP from inhibition of TP from E. ColiE. Coli
o Deprotection of the phosphonic acidsDeprotection of the phosphonic acids
•This method was applied to all phosphonic esters synthesized (fluorinated linear acyclonucleotides analogues and conformationally constrained)
Directed inhibition tests in collaboration with Dr. Murielle Muzard from the University of
Reims
McKenna, C. E. ; Higa, M. T. ; Cheung, N. H. ; McKenna, M.-C. Tetrahedron Lett. 1977, 2, 155-158.
P
OO
OF F
N( )n
NH
R
O
O
P
OHO
HOF F
N( )n
NH
R
O
O1/ TMSBr (5 eq.), CH2Cl2,
0 °C -> 1h, r.t. -> 72h
2/ MeOH, r.t., 2h
n = 3, 4R = H, Br, CH3
67-75%
Br
Br( )4
P(OiPr)3
130°C, 24hP
OO
OBr
81%
1/ N 3-Benzoylthmine,
TMG, DMSO, r.t., 16h
2/ MeNH2, MeOH, r.t., 16h (i-PrO)2P
O
92% 88%
( )4 N
HN OO
( )4
P
O
N
HN OO
( )4HOHO
o Non-fluorinated analogueNon-fluorinated analogue
Diab, S. A.; De Schutter, C.; Muzard, M.; Plantier-Royon, R.; Pfund, E.; Lequeux, T. J. Med. Chem. 2012, submitted.
330
10
20
30
40
50
60
70
1 2I (1mM)
P
F F
OHOHO
NH
N O
O
0
10
20
30
40
50
60
I (1mM)
• Increased activity with the fluorine compound CF2 > CH2
Influence of the fluorine atoms and the size of the spacerInfluence of the fluorine atoms and the size of the spacer
o Influence of the fluorine atomInfluence of the fluorine atom
• Long chain Better activity
% I = 51%
% I = 68%
o Influence of the size of the spacerInfluence of the size of the spacer
P
OHOHO
NH
N O
O
P
F F
OHOHO
NH
N O
O
P
F F
OHOHO
N
NH
O
O
% I = 51%
% I = 23%
I (1 mM)
I (1 mM)
Diab, S. A.; De Schutter, C.; Muzard, M.; Plantier-Royon, R.; Pfund, E.; Lequeux, T. J. Med. Chem. 2012, submitted.
34
Influence of the nucleobase and the iodine atomInfluence of the nucleobase and the iodine atom
o Influence of the nature of the nucleobaseInfluence of the nature of the nucleobase
o Influence of the iodine atomInfluence of the iodine atom
P
F F
OHOHO ( )3
N
NH
O
O P
F F
OHOHO ( )3
N
NH
O
O
Br
F F
P
O
HOHO
N
N
N
N
OH
NH2( )3
% I = 44% % I =
74%
% I = 61%
% I = 68%
P
F F
OHOHO
NH
N O
O
P
F F
OHOHO
NH
N O
O
I
% I = 95%
% I = 51%
I (1 mM)
I (1 mM)
P
F F
OHOHO ( )3
N
NH
O
O
• The presence of an iodine atom increase the activity
• Uracil Moderate activity
• Thymine, 5-bromouracil, guanine Good activity
P
F F
OHOHO
NH
N O
O
P
F F
OHOHO
NH
N O
O
I
% I = 52%
% I = 4%
I (100 M)
Diab, S. A.; De Schutter, C.; Muzard, M.; Plantier-Royon, R.; Pfund, E.; Lequeux, T. J. Med. Chem. 2012, submitted.
35
Influence of the traizoleInfluence of the traizole
o concentration of 1 mMconcentration of 1 mM
o Concentration of 100 Concentration of 100 MM
F F
P
OHOHO
NN
NN
N
N
N
OH
( )2
F F
P
OHOHO
NN
NN
N
N
N
OH
( )3
F F
P
OHOHO
NN
NN
N
N
N
OH
( )3NH2
F F
P
OHOHO
NN
NNHN
O
O
( )3
% I = 69%
% I = 76%
% I = 76%
% I = 29%
% I = 35%
% I = 35%
% I = 79%
% I = 90% % I =
79%
% I = 36%
% I = 68%
% I = 22%
• The activity increases with the size of the spacer• Best result obtained with compound conformationally constrained
F F
P
OHOHO
NN
NNHN
O
O
( )2
F F
P
OHOHO
NN
NNHN
O
O
( )3
I (1 mM)
I (100 M)
F F
P
OHOHO
NN
NNHN
O
O
( )3
Ki = 58 M
Diab, S. A.; De Schutter, C.; Muzard, M.; Plantier-Royon, R.; Pfund, E.; Lequeux, T. J. Med. Chem. 2012, submitted.
36
General conclusionGeneral conclusion
o Inhibitors of the family I: Linear chain Inhibitors of the family I: Linear chain
• Two synthetic athways: ionic and radical
P
OO
OF F
Base
I
NH
NH
O
O
NH
NH
O
O
NH
NH
O
O
Br
NH
N
N
N
Cl
NH2NH
N
N
N
Cl
( )n
n = 1, 471- 76%
P
F F
O
O
O
Base( )n
n = 1, 260- 78%
P
OO
OF F
O
Base
56- 62%
P
ORO
ROF F
4- 12 Å
Base( )n
P
ORO
ROF F
OH( )n
P
ORO
ROF F
P
ORO
ROI
F F
Base( )n+ P
ORO
ROF F
I onic process
Radical process
37
o Inhibitors with constrained conformation Inhibitors with constrained conformation
72%
NP
O
O
O
F F
HN
O
O
N
n = 0, 1, 2, 371- 96%
P
F F
OOO N
N
Base
( )n
NH
NH
O
O
NH
NH
O
O
NH
N
N
N
Cl
NH2NH
N
N
N
Cl
36%
ON
P
F FO
OO
HN
O
O
General conclusionGeneral conclusion
Base
( )n
P
ORO
ROF F
I onic process
POH
OOH
FF( )n
HO
( )nP
OR
OOR
FF( )n
P
ORO
ROI
F F+ P
ORO
ROF F
Radical process
38
ON
P
F FOH
OOH
HN
O
ON
P
O
HO
HO
F F
HN
O
O
P
OHO
HOF F
O
Base
BaseP
OHO
HOF F I
( )n
P
OHO
HOF F
Base( )n
OHP
OHO
HOF F
Base
OH
• « click chemistry » Inhibitors by autoselection of the enzyme
Conclusion and outlookConclusion and outlook
• All our phosphonic acids tested are active
P
OHO
HO NNH
O
O
% I (1 mM) = 68%% I (100 M) = 22%
Référence 1
Référence 2
• Test the other phosphonic acids.
• Design of new inhibitors long chain, fonctionnality, nucleic base
% I (1mM) = 90 %% I (100 M) = 68%Ki = 58 M
N NN
N
HNO OP
F FHO
OHO
P
OHO
HO NNH
O
Br
O
% I (1 mM) = 91%
% I (100 M) = 55%
Ki = 142 M
H2N
TP64
39
Post-doc work:DEAROMATIZATION OF NITROARENES BY
CYCLOADDITION
Supervisor: Dr. Isabelle CHATAIGNERLFAOC-IRCOF. UMR-CNRS 6014
Faculté des Sciences et Techniques. Rue Tesnières 76821 Mont Saint Aignan, Rouen. France
40
Dearomatization of Nitroarenes by cycloaddtionDearomatization of Nitroarenes by cycloaddtion
Subject I:Subject I: Dearomatization of Nitroarenes with Dearomatization of Nitroarenes with NN-Benzyl Azomethine Ylide -Benzyl Azomethine Ylide
Subject II:Subject II: Nitroarenes reactivity with enaminesNitroarenes reactivity with enamines
41
Dearomatization of Nitroarenes with Dearomatization of Nitroarenes with NN-Benzyl Azomethine Ylide -Benzyl Azomethine Ylide
Lee, S.; Chataigner, I.; Piettre, S. R. Angew. Chem. Int. Ed. 2011, 50, 472-476.
o Preliminary resultsPreliminary results
• Cycloaddition [3+2]
NO2
NO2
NO2
CO2Me
NO2
CF3
Cl
NO2
NO2
NO2N
NO2
NNO2N
p-tosN
N
N S
NO2
p-tos
NO2
p-tos
NO2
N
NO2
O
Reaction time: 0.5 to 26 h
Yield: 57 to 95%
TFA cat., CH2Cl2, 0°C à r.t., 4h
NO2
BnN TMSMeO
N
N Bn
BnNO2
NO2 NO2
BnN
85%+ H
H
2
1
42
• Bicycloadduct
• Regioisomer cis
o 1,2-dinitrobenzene1,2-dinitrobenzene
Dearomatization of Nitroarenes with Dearomatization of Nitroarenes with NN-Benzyl Azomethine Ylide -Benzyl Azomethine Ylide
o 3-chloronitrobenzene 3-chloronitrobenzene
TFA cat., CH2Cl2, 0°C to r.t., 24hBn
N35%
+
NO2
NO2N
N Bn
BnNO2
NO2
o 3,4-dichloronitrobenzene 3,4-dichloronitrobenzene
• The three possible chlorinated carbon-carbon double bonds were found to be unreactive
• Bicycloadduct
NO2
NO2
4h85%
trans bisadduct
NO2
4h69%
trans bisadduct
NO2
TFA cat., CH2Cl2, 0°C to r.t., 24hBn
N81%
+
NO2
Cl
N
N
Cl
O2NH
H
Bn
BnCl
Cl
Lee, S.; Diab, S.; Queval, P.; Chataigner, I.; Piettre, S. R. Angew. Chem. Int. Ed. 2012 (will be submitted).
BnN TMSMeO+
Cl
TFA cat., CH2Cl2, 0°C to r.t., 24h
NO2
33%
N
N
Cl
Bn
Bn H
O2N
H
H
43
Dearomatization of Nitroarenes with Dearomatization of Nitroarenes with NN-Benzyl Azomethine Ylide -Benzyl Azomethine Ylide
o 3,5-bis(trifluoromethyl)nitrobenzene3,5-bis(trifluoromethyl)nitrobenzene
TFA cat., CH2Cl2, 0°C to r.t., 1h
NO2 N
N Bn
Bn
NO2
NBn
NO2
NBn
+
CF3CF3
CF3F3C F3C
F3CBnN
98%+
30 / 70
+
NO2
N
N Bn
BnNO2
CF3
CF3
N
N Bn
Bn
O2N
CF3
TFA cat., CH2Cl2, 0°C to r.t., 24hBn
N
H
53%
H
H
H
NBn
NO2
CF3
NBn
NBn
O2N
N
CF3
Bn
+
o 3-(trifluoromethyl)nitrobenzene3-(trifluoromethyl)nitrobenzene
Lee, S.; Diab, S.; Queval, P.; Chataigner, I.; Piettre, S. R. Angew. Chem. Int. Ed. 2012 (will be submitted).
44
o Pyridine derivative: 3,5-dinitropyridinePyridine derivative: 3,5-dinitropyridine
Dearomatization of Nitroarenes with Dearomatization of Nitroarenes with NN-Benzyl Azomethine Ylide -Benzyl Azomethine Ylide
NN
O2N TFA cat., CH2Cl2, 0°C to r.t., 24hBn
N98%
+
NN
N
Bn
O2N H
o 5-nitro-1,10-phenanthroline5-nitro-1,10-phenanthroline
BnN TMSMeO+
N
NO2O2N TFA cat., CH2Cl2, 0°C à t.a., 1h
72% N
N
N
N
Bn
Bn
Bn NO2
O2N
Lee, S.; Diab, S.; Queval, P.; Chataigner, I.; Piettre, S. R. Angew. Chem. Int. Ed. 2012 (will be submitted).
CNNC
CNNC
CO2MeMeO2C
CO2MeMeO2C
CNCl
CNClCO2Me
CF3
F3C
o Substrats without nitro groupSubstrats without nitro group
45
Subject I:Subject I: Dearomatization of Nitroarenes with Dearomatization of Nitroarenes with NN-Benzyl Azomethine Ylide -Benzyl Azomethine Ylide
Subject II:Subject II: Nitroarenes reactivity with enaminesNitroarenes reactivity with enamines
Dearomatization of Nitroarenes by cycloaddtionDearomatization of Nitroarenes by cycloaddtion
NO
OR1
[4+2]
ON
O
R1**
FunctionalizationO
N
OH
R1**
R2
*OH
NH2
R1**
R2
*
N-O bond
cleavage
[3+2]
ON
R1**
O
R1
*
46
Nitroarenes reactivity with enaminesNitroarenes reactivity with enamines
o The aims of the projectThe aims of the project
• Re-evaluating the synthetic potential of nitroaromatic compounds
• Dearomatization
• nitronate chemistry
N
O
• Enamine : 4-cyclohexenylmorpholine
O
OOH
NO2
N
O
+ CH2Cl2, r.t., 3h
65%
O
O O
NO2
(3 eq.)
N
O
47
Nitroarenes reactivity with enaminesNitroarenes reactivity with enamines
o Reaction at atmospheric pressure (unpublished)Reaction at atmospheric pressure (unpublished)
N
NO2
Ts
N
O
+CH2Cl2, r.t., 30 min
77%N
NO2
Ts
N
O
(3 eq.)
NitroindoleNitroindole
NitrobenzofuranNitrobenzofuran
• Mild conditions of dearomatization
• Unstable product
• Furan ring opening
NO2
Cl
ClN
NO2NO2
NO2
NO2
N
NO2
N
O2NNO2
NO2
N
NO2
• No reaction
48
o Reaction under high pressure (unpublished)Reaction under high pressure (unpublished)
Nitroarenes reactivity with enaminesNitroarenes reactivity with enamines
N
O
14kbar
+CH2Cl2, r.t., 24h
NOO
ON
O
N
O
Conjugate
addition
NOO
N
O
Nitronate intermediate
ON
O N
O
Rearomatization
Hydrolysis of iminium
HOH
NO O
Formation ofnitroso
ON
ON
O
N OO
N
O
49
o Reaction under high pressure (unpublished)Reaction under high pressure (unpublished)
Nitroarenes reactivity with enaminesNitroarenes reactivity with enamines
N
O
14kbar
+CH2Cl2, r.t., 24h
NOO
ON
O
N
O
N
ON
O
N
O
87%
ON
O
N
O
75%
48% N
NO
O
N
O
ON ON
O
O
O
O
51%
50
o Reduction: Cleavage of the N-O bond (unpublished)Reduction: Cleavage of the N-O bond (unpublished)
Nitroarenes reactivity with enaminesNitroarenes reactivity with enamines
• PtO2, H2 NO
• Ni/Raney, 10 bar H2 NO
• H-Cube trace
Under acidic conditionUnder acidic condition
ON
O
N
O
Ni/Raney, H2
MeOH, 48h, r.t.
ON
O
O
X
ON
O
N
O
X
NO
TFAcat.
CH2Cl2, r.t., 30min.
X = CH 88%X = N 91%
51
ConclusionConclusion
Dearomatization of Nitroarenes with Dearomatization of Nitroarenes with NN-Benzyl Azomethine Ylide -Benzyl Azomethine Ylide
• Uncomplexed benzene derivatives acting as 2 components in an intermolecular pericyclic process.
• The formed products, functionnalized, constitute scaffolds prone to a potentially interesting derivatisations.
• Loss of aromaticity of nitrobenzene and other nitroarenes.
Nitroarenes reactivity with enaminesNitroarenes reactivity with enamines
• We have discovered a new type of reactivity of enamine under high pressure that allows the rapid generation of polycyclic compounds
• Applicable to different nitroarenes
• In only two steps, starting from simple compounds, original polycyclic aminated compounds are efficiently obtained
