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Group Meeting Montreal, February 22, 2012 Sonia Diab. CONSTANTINE. CAEN (Basse-Normandie. ROUEN (Haute-Normandie). SYNTHESIS OF DIFLUOROPHOSPHONATES BY CARBANIONIC AND RADICAL PROCESS FOR THE PREPARATION OF THYMIDINE PHOSPHORYLASE INHIBITORS. Sonia DIAB Montreal, February 22, 2012.
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SYNTHESIS OF DIFLUOROPHOSPHONATES BY CARBANIONIC AND RADICAL PROCESS FOR THE PREPARATION OF THYMIDINE PHOSPHORYLASE INHIBITORS Sonia DIAB Montreal, February 22, 2012 PhD Supervisor: Pr. Thierry. LEQUEUX Laboratoire de Chimie Moléculaire et Thio-organique. UMR-CNRS 6507 6 Boulevard du Maréchal Juin 14050 CAEN, France
Introduction • Tumoral Angiogenesis PD-ECGF::Platelet-derived endothelial cell growth factor • Protein involved in angiogenesis of cancerous cells. • Identified as Human Thymidine Phosphorylase (HTP) (a) Folkman J and al. J Biol Chem1992 ; 267 : 10931-10934, (b) Ishikawa, F. and al. Nature 1989, 338, 557-562. • Propperties of Human Thymidine Phosphorylase Inhibitor of TP (a) Walter, M. R. And al. J. Biol. Chem. 1990, 265, 14016-14022. (b) Norman, R. A. And al. Structure2004, 12, 75-84.
Inhibitors of Thymidine phosphorylase • Multisubstrats Inhibitors • Very few TP inhibitors described • Phosphonate as mimics of the phosphate • function Blazarini, J. and al. J. Med. Chem.2000, 43, 971-984. • Analogie phosphate / phosphonate / fluorophosphonate • Difluorophosphonates best isopolar and isosteric phosphate analogue Blackburn, M. G. ; England, D. A. ; Kolkmann, F. J. Chem. Soc. Chem. Comm. 1981, 930-392 • No fluorinates TP inhibitors described to date Synthesis of new fluorinated inhibitors of TP
Les Families of TP Inhibitors • Family I : Linear Inhibitors • Family II : Conformationally constrained Inhibitors How to introduce the difluoromethylphosphonate function
General 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. Synthesis1996, 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) Blades, K. et al. Chem. Commun. 1996, 1457-1458. (b) Xu, Y. et al. Org. Lett. 2002, 4, 4021-4024. (a) Solas, D. et al. J. Org. Chem. 1996, 61, 1537-1539. (b) Lequeux, T. et al. Org. Lett.2001, 3, 185-188
Limits: based on the use of HCFC or CFC (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. • 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.
Previous work of the laboratory • New method to generate the anion • Fluorosulfide: Novel source of the anion developed in Lequeux’s Laboratory Henry-dit-Quesnel, A. ; Toupet, L. ; Pommelet, J. C. ; Lequeux, T. Org. Biomol. Chem. 2003, 1, 2486-2491 • Study of th reactivity of the anion depending on its source • E+ : TMSCl, aldehyde, ketone, CS2 The same reactivity • E+ : , , , DMF Different reactivity Ozouf, P. ; Binot, G. ; Pommelet, J. C. ; Lequeux, T. Org Lett.2004, 6, 21, 37-47. Anion can open oxacycles to prepare corresponding alcohols.
Family I: Inhibitors with a linear chain
Hydroxyphosphonates: Key molecules for the synthesis of our inhibitors • Formation of hydroxydifluoromethylphosphonates with various size • Opening reations of oxacycles
Preparation of b-hydroxydifluoromethylphosphonates • 1,2-cyclic sulfates opening Diab, S. A. ; Sene, A. ; Pfund, E. ; Lequeux, T. Org. Lett. 2008, 10, 3895-3898.
Preparation of g-hydroxydifluoromethylphosphonates • Oxetanes opening Ozouf, P. ; Binot, G. ; Pommelet, J. C. ; Lequeux, T. Org Lett.2004, 6, 21, 37-47. Diab, S. A. ; Sene, A. ; Pfund, E. ; Lequeux, T. Org. Lett. 2008, 10, 3895-3898.
Preparation of d-hydroxydifluoromethylphosphonates • THF opening Ozouf, P. ; Binot, G. ; Pommelet, J. C. ; Lequeux, T. Org Lett.2004, 6, 21, 37-47. • THP opening • No reaction • The size of oxacycles opening is limited to the THF Exploit other alternatives to increase the size of the spacer
Preparation of w-hydroxydifluoromethylphosphonates • Lactones opening: e-caprolactone and z-lactone • Lactones opening leads to the formation of hydroxyketodifluoromethylphosphonates Introduction of the nucleic base
Preparation of fluorinated analogues of linear acyclonucleotides • Approach 1: Mitsunobu Reaction • Direct introduction of the nucleic base by Mitsunobu reaction Moderate yields and nonreproducible reaction • Approach 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 Using TMG to introduce the nucleic base Diab, S. A. ; Sene, A. ; Pfund, E. ; Lequeux, T. Org. Lett. 2008, 10, 3895-3898.
Preparation of fluorinated analogues of linear acyclonucleotides • Introduction of the nucleic base Diab, S. A. ; Sene, A. ; Pfund, E. ; Lequeux, T. Org. Lett. 2008, 10, 3895-3898
Access to long-chain inhibitors • Synthesis according to the first strategy: ionic process • Early introduction of the difluoromethylphosphonate group • Late introduction of the nucleobase • Synthesis according to the second strategy: radical process • Late introduction and at the same time of the difluoromethylphosphonate group and the nucleobase
Radical reaction • Preparation of iododifluoromethylphosphonate • Chalcogen-halogen exchange avoids the use of HCFCs Henry-dit-Quesnel, A. ; Toupet, L. ; Pommelet, J. C. ; Lequeux, T. Org. Biomol. Chem. 2003, 1, 2486-2491 • Reaction with n-octene • Three initiator systems were used • Regioselective reaction Sène, A. ; Diab, S. ; Hienzsch, A. ; Cahard, D. ; Lequeux, T. Synlett. 2009, 981-985. Functionalized alkenes
Addition of the radical phosphonodifluoromethyle on the alkenes • Addition on the allylthymine • Addition on functionalized alkenes with long-chain • Reaction performed in the presence of purine and pyrimidine base
Family II: Conformationally constrained Inhibitors
Retrosynthsis 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
Conformation frozen by a furan ring • Radical addition on 2,3-dihydrofuran • 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
Conformation frozen by a furan ring • Introduction of the nucleic base • Diastereoselective reaction (trans configuration) • Formation of two regioisomers N 1 and N 3
Conformation frozen by a benzene ring • Preparation of 1,4-cyclic sulfate (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. • 1,4-cyclic sulfate opening • First exemple of 1,4-cyclic sulfate opening by difluoromethylphosphonate anion
Conformation frozen by a triazole ring • 1,3-dipolar cycloaddition Diab, S. A. ; Hienszch, A. ; Lebargy, C. ; Pfund, E. ; Lequeux, T. Organic and Biomolecular Chemistry, 2009, 21, 4481-4490.
Tests of inhibition of Thymidine phosphorylase from E.Coli
In vitro inhibition of TP from E. Coli • Deprotection of the phosphonic acids McKenna, C. E. ; Higa, M. T. ; Cheung, N. H. ; McKenna, M.-C. Tetrahedron Lett. 1977, 2, 155-158. • This method was applied to all phosphonic esters synthesized (fluorinated linear acyclonucleotides analogues and conformationally constrained) • Non-fluorinated analogue Directed inhibition tests in collaboration with Dr. Murielle Muzard from the University of Reims Diab, S. A.; De Schutter, C.; Muzard, M.; Plantier-Royon, R.; Pfund, E.; Lequeux, T. J. Med. Chem. 2012, submitted.
Influence of the fluorine atoms and the size of the spacer • Influence of the fluorine atom • Increased activity with the fluorine compound • CF2 > CH2 % I = 23% % I = 51% I (1 mM) • Influence of the size of the spacer % I = 68% % I = 51% • Long chain Better activity I (1 mM) Diab, S. A.; De Schutter, C.; Muzard, M.; Plantier-Royon, R.; Pfund, E.; Lequeux, T. J. Med. Chem. 2012, submitted.
Influence of the nucleobase and the iodine atom • Influence of the nature of the nucleobase % I = 61% % I = 44% % I = 74% % I = 68% • Uracil Moderate activity • Thymine, 5-bromouracil, guanine Good activity I (1 mM) • Influence of the iodine atom • The presence of an iodine atom increase the activity % I = 95% % I = 52% % I = 51% % I = 4% I (1 mM) I (100 mM) Diab, S. A.; De Schutter, C.; Muzard, M.; Plantier-Royon, R.; Pfund, E.; Lequeux, T. J. Med. Chem. 2012, submitted.
Influence of the traizole • concentration of 1 mM % I = 79% % I = 90% % I = 76% % I = 79% % I = 76% % I = 69% I (1 mM) • Concentration of 100 mM % I = 68% % I = 36% % I = 35% % I = 35% % I = 22% % I = 29% • The activity increases with the size of the spacer • Best result obtained with compound conformationally constrained I (100 mM) Diab, S. A.; De Schutter, C.; Muzard, M.; Plantier-Royon, R.; Pfund, E.; Lequeux, T. J. Med. Chem. 2012, submitted.
General conclusion • Inhibitors of the family I: Linear chain • Two synthetic athways: ionic and radical
General conclusion • Inhibitors with constrained conformation
Conclusion and outlook • All our phosphonic acids tested are active Référence 1 Référence 2 • « click chemistry » Inhibitors by autoselection of the enzyme • Test the other phosphonic acids. • Design of new inhibitors long chain, fonctionnality, nucleic base
Post-doc work:DEAROMATIZATION OF NITROARENES BY CYCLOADDITION Supervisor: Dr. Isabelle CHATAIGNER LFAOC-IRCOF. UMR-CNRS 6014 Faculté des Sciences et Techniques. Rue Tesnières 76821 Mont Saint Aignan, Rouen. France
Dearomatization of Nitroarenes by cycloaddtion Subject I:Dearomatization of Nitroarenes with N-Benzyl Azomethine Ylide Subject II:Nitroarenes reactivity with enamines
Dearomatization of Nitroarenes with N-Benzyl Azomethine Ylide • Preliminary results • Cycloaddition [3+2] Lee, S.; Chataigner, I.; Piettre, S. R. Angew. Chem. Int. Ed. 2011, 50, 472-476.
Dearomatization of Nitroarenes with N-Benzyl Azomethine Ylide • 1,2-dinitrobenzene • Bicycloadduct • Regioisomer cis • 3-chloronitrobenzene • 3,4-dichloronitrobenzene • The three possible chlorinated carbon-carbon double bonds were found to be unreactive • Bicycloadduct Lee, S.; Diab, S.; Queval, P.; Chataigner, I.; Piettre, S. R. Angew. Chem. Int. Ed. 2012 (will be submitted).
Dearomatization of Nitroarenes with N-Benzyl Azomethine Ylide • 3-(trifluoromethyl)nitrobenzene • 3,5-bis(trifluoromethyl)nitrobenzene Lee, S.; Diab, S.; Queval, P.; Chataigner, I.; Piettre, S. R. Angew. Chem. Int. Ed. 2012 (will be submitted).
Dearomatization of Nitroarenes with N-Benzyl Azomethine Ylide • Pyridine derivative: 3,5-dinitropyridine • 5-nitro-1,10-phenanthroline • Substrats without nitro group Lee, S.; Diab, S.; Queval, P.; Chataigner, I.; Piettre, S. R. Angew. Chem. Int. Ed. 2012 (will be submitted).
Dearomatization of Nitroarenes by cycloaddtion Subject I:Dearomatization of Nitroarenes with N-Benzyl Azomethine Ylide Subject II:Nitroarenes reactivity with enamines
Nitroarenes reactivity with enamines • The aims of the project • Re-evaluating the synthetic potential of nitroaromatic compounds • Dearomatization • nitronate chemistry • Enamine : 4-cyclohexenylmorpholine
Nitroarenes reactivity with enamines • Reaction at atmospheric pressure (unpublished) Nitroindole • Mild conditions of dearomatization • Unstable product Nitrobenzofuran • Furan ring opening • No reaction
Nitroarenes reactivity with enamines • Reaction under high pressure (unpublished)
Nitroarenes reactivity with enamines • Reaction under high pressure (unpublished)
Nitroarenes reactivity with enamines • Reduction: Cleavage of the N-O bond (unpublished) • PtO2, H2 NO • Ni/Raney, 10 bar H2 NO • H-Cube trace Under acidic condition