Institut Curie / CNRS UMR 176 Conception, Synthesis and Targeting
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Institut Curie / CNRS UMR 176 Conception, Synthesis and Targeting
Institut Curie / CNRS UMR 176 Conception, Synthesis and Targeting of Biomolecules Unit director: Jean-Claude Florent, PhD The central activity of this chemistry unit is to develop and use small molecule ligands to control the biological activities of important target proteins. and nucleic acids. Organic and Organometallic synthesis is being used to design new bioactive compounds, as well as natural-product analogues, new chemicals to cross-link proteins and new probes of biological function. Currently, the Institut Curie–CNRS's proprietary library is comprised of 9 000 small chemical compounds arrayed in 96-well plates. The main research themes of the unit include: Antivascular compounds (such as tubulin-binding ligands). Kinase inhibitors G-quadruplex targeting agents DNA fluorescent dyes Photodynamic therapy of tumors with new porphyrines. Antitumour drug-targeting compounds for antibody-directed enzyme prodrug therapy Shiga toxin as a vector for agent delivery (molecules, siRNA, immunogenic petides) Methodology development in organic and organometallic chemistry. Protein–protein interaction inhibitors Jean-Claude Florent, PhD Medicinal Chemistry, Chemical biology, Drug Targeting Institut Curie / CNRS UMR 176 26 rue d'Ulm 75248 Paris Tel : +33 1 56 24 66 58 Fax : +33 1 56 24 66 31 Email : [email protected] KEYWORDS : Antivascular agents, kinase inhibitors, drug targeting, chemical probes for biology, anti-cell migrating agents, libraries of small molecules, methyl-transferase inhibitors, tubulin agents, CK2 inhibitors, Phosphatase inhibitors The main activity of my “Medicinal Chemistry, Bioorganic Chemistry, Targeting" group is the pharmacomodulation and synthesis of small molecules for the study of living organisms. The synthesized molecules find multiple applications as probes for proteomics/genomics and/or as agents for therapeutic purposes. We have a strong research activity on antitumour drug targeting, particularly through the use of Shiga toxin-drug conjugates (antitumour) or Shiga-peptide conjugates (for antitumour immunology). Our laboratory is also involved in another important field,, i.e. targeted therapy. Thus, we particularly develop flavonoid compounds as aminopeptidases inhibitors, potentially anti-angiogenic and anti-metastastic. We have also synthesized tubulin ligands for their potential antivascular activity, in particular analogues of Combretastatin A4, and have conceived serine threonine kinases, inhibitors of CK2 and PIM1. We have organized the Curie-CNRS “chimiothèque”, a collection of small molecules, comprising the products synthesized at UMR 176, to which we continue adding small molecules. Some of the hits found by our biologists partners are being optimized as inhibitors of protein-protein interactions involved in cell migration and metastasis (Llamiline/Syndecan3) and we develop a hit inhibitor of the serine-threonine phosphatase I (PP1). Furthermore, within a Curie network in connection with a team of biologists from our Institute, we synthesize chemical probes to study molecular biology and mechanisms relating to the retrograde route taken by the Shiga toxin to the Golgi apparatus. This chemical biology programme has allowed us to synthesize chemical probes applied for the study of the Shiga toxin endocytosis involving cell fluorescence microscopy on living cells or on Geant Unilamelar vesicles as cell models membrane. We have undertaken a programme on the proteomic analysis of the cell surface proteins, which could follow this retrograde route of endocytosis. Figure 1 : Shiga toxin B subunit as a vector for antitumour drugs The Shiga toxin B subunit (STxB) is used for intracellular retrograde delivery of drug. Recent/Key Publications A. El Alaoui et al. (2008) Synthesis and properties of a mitochondrial peripheral benzodiazepine receptor conjugate. ChemMedChem. 3, 1687-1695. R. Prudent et al. (2008) Expanding the chemical diversity of CK2 inhibitors. Mol Cell Biochem, 316, 71-85. W. Römer et al. (2007) Shiga toxin induces tubular membrane invaginations for its uptake into cells. Nature 450, 670-675. A. El Alaoui et al. (2007) Shiga toxin-mediated retrograde delivery of a topoisomerase I inhibitor prodrug. Angewandte Chemie 119, 6469-6472. M. Arthuis, R. Pontikis, J.-C. Florent. (2009) Stereoselective Synthesis of Novel Highly Substituted Isochromanone and Isoquinolinone-Containing Exocyclic Tetrasubstituted. Alkenes J. Org. Chem. 74, 2234-2237. Patents available for licensing : 5 Philippe Belmont, PhD Organometallic chemistry, heterocycles and biological targets Institut Curie / CNRS UMR 176 26 rue d'Ulm 75248 Paris Cedex 05 Tel : +33 1 56 24 68 24 Fax : +33 1 56 24 66 31 Email : [email protected] KEYWORDS : Heterocyclic chemistry, cycloisomerization and tandem reactions, silver-catalyzed and goldcatalyzed transformations, protein-kinase inhibitors, anti-proliferative and antipaludic properties. Our research group deals with an original access to novel heterocyclic structures by organometallic and organic chemistry. Then, their biological targets and anticancer properties are investigated. Our first project allowed us to access the acridine scaffold, a known bioactive heterocycle, through a benzene ring formation (benzannulation, Fig. 1), thanks to an ene-yne cycloisomerization reaction catalyzed with rhodium(I) salts ([Rh] cat.). We then looked for a more general method, and although the cationic gold complex Au(PPh3)SbF6 proved to be the most active specie for this transformation, silver salts such as AgSbF6 (needed for the anion metathesis in the gold chemistry) led us to develop an original cycloisomerization reaction for the access to polyaromatic derivatives: naphthalenes, acridines and quinolines (Fig.1). O OTBS Y R Y = N or CH OTBS or [Ag] cat. Y O N R2 R2 Furoquinoline Pyranoquinoline Figure 2: Tandem acetalization/heterocyclization reaction. Finally, we have synthesized tetrahydrocyclopenta[c]acridinone structures in only 3 steps from a commercially available quinoline. The key chemical step is a Pauson-Khand reaction leading to an active compound on cyclin-dependent kinases at submicromolar concentration (Fig. 3, kinase inhibition). O Key step : Pauson Khand reaction Cl Commercially available quinoline R2 N R3 O tetrahydrocyclopenta[c]acridinones Heterocycle (in orange) Kinase ATP binding pocket (in green) R Y = CH : Naphthalenes Y = N : Acridines, quinolines N R2 N [Rh] cat. or [Au] cat. R1 OR3 OR3 O or M = [Ag], [Au] cat. N H BENZANNULATION R1 Base (K2CO3) or R1 Figure 3: Kinase inhibition. AMINOBENZANNULATION R1 O R1 n R N R2 Recent/Key Publications n Y Y Starting material N H R2 R n =1, Y = N : Acridines, quinolines n = 0, Y = NR : Carbazoles n = 0, Y = O : Dibenzofurans Figure 1: Benzannulation and aminobenzannulation reactions. With the same starting material, thanks to the use of a secondary amine, the enamine intermediate triggers the cycloisomerization reaction onto the alkynyl group (aminobenzannulation) to obtain bioactive heterocycles such as acridines, quinolines, carbazoles and also dibenzofurans (Fig. 1). Also, our work is directed towards the access to furoquinoline and pyranoquinoline cores, structurally related to known bioactive alkaloids. First of all, the reaction was developed via a base-catalyzed tandem cycloisomerization reaction and thereafter a more general method was developed by organometallic catalysis with gold and various silver salts. More importantly, depending on the silver salt used (Ag2O versus AgOTf) a regioselective 5-exo-dig versus 6-endo-dig cyclization was observed, leading respectively to furoquinolines and pyranoquinolines (Fig. 2). P. Belmont et al. (2009). Silver and Gold Catalysis for Cycloisomerization Reactions. Eur. J. Org. Chem., 6075-6089. M. Tiano et al. (2008). Rapid access to amino-substituted quinoline, (di)benzofuran and carbazole heterocycles through an aminobenzannulation reaction. J. Org. Chem., 73, 41014109. T. Godet et al. (2007). Silver versus Gold Catalysis in Tandem Reactions of Carbonyl Functions onto Alkynes: a Versatile Access to Furoquinoline and Pyranoquinoline Cores. Chem. Eur. J., 13, 5632-5641. P. Belmont et al. (2007). Acridine and Acridone Derivatives, Anticancer Properties and Synthetic Methods: Where Are We Now? Anti-Cancer Agents Med. Chem., 7, 139-169. P. Belmont et al. (2005). An Efficient and Simple Aminobenzannulation Reaction: Pyrrolidine as a Trigger for the Synthesis of 1-Amino-Acridines. Org. Lett., 7, 1793-1795. Marie-Paule Teulade-Fichou, PhD Heterocyclic Chemistry and Design of Biomolecules Interactions Inhibitors Institut Curie / CNRS UMR 176 Centre Universitaire 91405 Orsay Cedex Tel : +33 1 69 86 30 86 Fax : +33 1 69 07 53 81 Email : [email protected] KEYWORDS : Heterocyclic chemistry, cancer therapeutic agents, quadruplex nucleic acids ligands, telomere targeting agents, DNA fluorescent dyes, retinoblastoma, photosensitising molecules, compound library screening, hit identification and optimisation Our group works on the design of compounds that target various proteins, enzymes and nucleic acids of interest for cancer therapy. Our research is carried out in close collaboration with biochemists, biologists and clinicians. The group has a broad expertise in organic synthesis and bioorganic chemistry with a strong background in molecular recognition of biological targets. mouse model grafted with an anatomopathological sample of human retinoblastoma. The Institut Curie-CNRS compound library To date our diverse chemical libraries is composed of more than 8500 compounds. Our collection has been screened against a number of therapeutically interesting targets which led to the identification of promising novel chemical hits. Some of DNA structural probes these are currently being optimised, and in particular some We are designing probes with the property to bind to protein-protein interaction modulating agents. secondary structures of DNA in strategic regions of the genomic DNA. For example we are actively working at developing distinct families of G-quadruplex DNA binding molecules and Mismatched DNA ligands which could offer new perspectives in anticancer drug discovery (Fig. 1). In addition, we have recently developed a novel family of redemitting dyes for use in optical tracking of DNA in cells by multiphoton microscopy. Figure 2: The structure of glyco-conjugated porphyrin-based photosensitisers Recent/Key Publications Figure 1 : Structure of a quinacridine ligand and of its complex with G-quadruplex DNA determined by NMR. Purine libraries We are constructing a library of novel and diverse purinebased compounds to be used to develop inhibitors of molecular targets in Alzheimer's disease, Huntington's disease (e.g. GSK-3, CDK5, CK1) and cancer (Tyrosine kinases). The receptor tyrosine kinase c-Kit for example is known to be involved in hematopoiesis and to be overexpressed in various cancerous pathologies. In collaboration with an industrial partner, we are developing specific inhibitors of c-Kit and its mutated form. Piazza A. et al. (2010) Genetic instability triggered by Gquadruplex interacting Phen-DC compounds in Saccharomyces cerevisiae Nucleic Acid.Res. published on line March 19th Brevet, D. et al. (2009) Mannose-targeted mesoporous silica nanoparticles for photodynamic therapy. Chem. Commun. 1475-1477. Granzhan A; et al. (2009) Macrocyclic DNA-Mismatch-Binding Ligands: Structural Determinants of Selectivity. Chem.Eur.J.,15,1314 Yang P. et al. (2009) Engineering Bisquinolinium-Thiazole orange conjugates for fluorescent sensing of G-quadruplex DNA Angew. Chem. Int .Ed. 48, 2188-2191. Keriel A. et al. (2009) Protection against Retrovirus Pathogenesis by SR Protein Inhibitors PLoS One 2009, 2, 4, e4533. Besselièvre F. et al. (2009) Copper-Catalyzed Direct Alkynylation of Azoles. Angew. Chem. Int. Ed. Engl. 48, 9553- 9556. Photosensitising molecules for retinoblastoma The current treatment for retinoblastoma, the most frequent eye tumour in children, exposes the patient to the long-term Patent available for licensing : 1 consequences of chemotherapy. The development of alternative treatment using non-mutagenic photosensitizers (i.e. compounds that damage cell components upon photoactivation) is of particular interest for treating this tumour type. The photosensitizers developed in our laboratory are glycoconjugated tetrapyrrole macrocycles (Fig. 2). We are in the process of evaluating the in vivo efficacy of these glycoconjugated photosensitisers in an immunodeficient