New Variety of Pyridine and Pyrazine-Based Arginine Mimics : Synthesis, Structural Study and Preliminary Biological Evaluation Olga Ovdiichuk To cite this version: Olga Ovdiichuk. New Variety of Pyridine and Pyrazine-Based Arginine Mimics : Synthesis, Structural Study and Preliminary Biological Evaluation. Chemical and Process Engineering. Université de Lorraine, 2016. English. NNT : 2016LORR0289. tel-01540911 HAL Id: tel-01540911 https://tel.archives-ouvertes.fr/tel-01540911 Submitted on 16 Jun 2017 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. 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Contact : [email protected] LIENS Code de la Propriété Intellectuelle. articles L 122. 4 Code de la Propriété Intellectuelle. articles L 335.2- L 335.10 http://www.cfcopies.com/V2/leg/leg_droi.php http://www.culture.gouv.fr/culture/infos-pratiques/droits/protection.htm Doctoral School RP2E Science and Engineering, Resources, Processes, Products and Environment New Variety of Pyridine and Pyrazine-Based Arginine Mimics: Synthesis, Structural Study and Preliminary Biological Evaluation A dissertation presented to the Université de Lorraine and the Taras Shevchenko National University of Kyiv To obtain the degree Doctor of Philosophy Olga OVDIICHUK Jury members Florine CAVELIER DR, IBMM, Université de Montpellier, Reviewer Claude TAILLEFUMIER Prof., Université Clermont-Ferrand, Reviewer Marc LE BORGNE Prof., Université Claude Bernard Lyon 1, Examinator Zoia VOITENKO Prof., Taras Shevchenko National University of Kyiv, Examinator Alain DURAND, Prof., Université de Lorraine, Examinator Marie-Christine AVERLANT-PETIT CR, LCPM, Université de Lorraine, Director Olga HORDIYENKO Dr., Taras Shevchenko National University of Kyiv, Co-director Axelle ARRAULT, MCF, Université de Lorraine, guest member, Co-director November 28, 2016 ABSTRACT Heterocycle-containing amino acids have become important precursors in the design of peptidomimetics. The pyridine and pyrazine rings can improve the binding affinity and bioavailability of potential drugs. Moreover, an enzymatic degradation resistance is known to be more pronounced in peptidomimetics with heterocyclic cores as non-peptidic fragments. In addition, an amidoxime group introduced into the structure represents potent pharmacophore due to the ability for its in vivo oxidation to amide with a subsequent release of NO (antiaggregatory activity) or their reduction to amidine, an excellent mimic of the arginine side chain. The use of amidoximes as amidine prodrugs allows to overcome unfavorable physicochemical and pharmacokinetic properties. This work hence describes the synthesis, structural study and preliminary biologycal evaluation of new variety of pyridine and pyrazine-based peptidomimetics. The first part of the thesis is devoted to the design and convenient synthesis of novel peptidomimetics bearing amidoxime function. Moreover, the introduction of an additional amino acid through different linkage like hydrazide, ester or heterocyclic unit (1,2,4-oxadiazole, 1,2,4-triazole) was of our great interest. 1,2,4-oxadiazole and 1,2,4-triazole rings are known as isosteric replacement of the amide and/or ester group due to its high resistance to metabolic degradation that has been widely used in peptide mimicry. Thus, several chemical functionalization of new scaffolds were studied. We have developed the acylation of amidoximes with further microwave- assisted condensation into amino acid derived 1,2,4-oxadiazoles. The synthesis of 1,2,4- triazoles was performed via N-acylamidrazones. The latter were synthesized in mild conditions using a new approach from the pyrrolopyridines(pyrazines) precursors in good to excellent yields. On further developing constrained peptidomimetics, we also synthesized hydrazide modified turn mimics derived from amidoximes. The second section describes structural analysis of the prepared compounds by NMR, IR spectroscopic studies and molecular modelling. Crystal structures of some compounds were analyzed by X-Ray diffraction study. Conformational preferences and thermodynamic studies of proline-containing peptidomimetics were investigated and both series – pyridine and pyrazine ones were compared. Examination of a new ProPhe pyrazine-based pseudotripeptide revealed the hydrogen bond formation between the proton of the OH and the carbonyl oxygen of the C-terminal phenylalanine and the hydrogen bond that adopts a seven- membered -turn conformation. Therefore, a dramatic increase of the trans rotamer up to 98% 2 was observed in weakly polar solvent, which is CHCl3. Hydrazide modified peptidomimetics adopt a turn structure in solution also stabilized by the hydrogen bond forming C10- pseudocycle. Conformational studies confirmed that these heterocyclic moieties can be used to increase rigidity and the pyrazine core could stronger effect on conformation stabilization. The third part of the thesis is dedicated to the preliminary results of the NO release assay on amidoximes. All compounds revealed release NO in concentration sufficient for pharmaМologiМal ОffОМts (≥1 µM). A tОntativО МorrОlation bОtwООn struМturО anН aМtivity was performed. 3 DEDICATION To my family 4 ACKNOWLEDGMENTS I would like to thank everybody who contributed to the success of this work. First and foremost, I would like to thank Dr. Olga V. HORDIYENKO, Dr. Axelle ARRAULT and Dr. Marie-Christine AVERLANT-PETIT for the opportunity to work on these interesting project. Their mentorship and the insightful discussions have been a major encouragement during my studies. I would also like to thank the members of the jury Prof. Claude TAILLEFUMIER, Dr. Florine CAVELIER, Prof. Marc LE BORGNE and Prof. Zoia VOITENKO for accepting the invitation to serve on my committee. Additionally, I thank Dr. Caroline GAUCHER at the Faculty of Pharmacy of the Université de Lorraine for the collaboration on the NO-donors project and the biological testing of my compounds. I also thank the Taras Shevchenko National University of Kyiv and the Campus France for financial support of this work and the Université de Lorraine for ATER position which gave me the opportunity to finish my thesis in four years. My thanks also go to the members of LCPM for their suggestions and a pleasant working atmosphere: Prof. Alain DURAND, Dr. Jacques BODIGUEL, Dr. Regis VANDERESSE, Dr. Guillaume PICKAERT, Dr. Samir ACHERAR, Dr. Olivier FABRE and Mme Mathilde ACHARD. Furthermore, I would like to thank Dr. Emmanuel WENGER (CRM2), Dr. Volodymyr MEDVIEDIEV and Prof. Oleg SHISHKIN (Institute for Single Crystals, NASU) for their X-ray diffraction measurements. I am especially thankful for Tanya SAHYOUN and Mohamed IBRAHIM with whom I shared the lab and to Amirah GAZZALI, they always made me looking forward to come here and were always willing to show support. Most importantly, I am extremely grateful for my parents, who have always supported me and encouraged me in my education. Equally important was the emotional support from my sister Alina and my friends Viktoriia, Iulia, Anastasiia, Vika and Larisa; I deeply appreciate your friendship. Thank you / Merci beaucoup / Щио ку! Olga 5 TABLE OF CONTENTS Abstract ...................................................................................................................................... 2 Dedication .................................................................................................................................. 4 Acknowledgments ...................................................................................................................... 5 Table of Contents ....................................................................................................................... 6 Abbreviations ............................................................................................................................. 8 Chapter 1: Introduction ............................................................................................................ 11 1. Introduction .................................................................................................................. 12 2. Pyridine and pyrazine heterocycles as scaffolds for peptidomimetics ......................... 12 3. Amidoximes and masked amidoximes as prodrugs of amidines, an arginine mimics . 13 4. Amidoximes as NO donors .......................................................................................... 19 Chapter 2: Synthesis of Pyridine and Pyrazine-based Peptidomimetics .................................. 23 1. Introduction .................................................................................................................