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Synthèse Et Evaluation De Nouveau Agents De Protection Contre Les Rayonnements Ionisants Brice Nadal

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Synthèse Et Evaluation De Nouveau Agents De Protection Contre Les Rayonnements Ionisants Brice Nadal Synthèse et Evaluation de nouveau agents de protection contre les rayonnements ionisants Brice Nadal To cite this version: Brice Nadal. Synthèse et Evaluation de nouveau agents de protection contre les rayonnements ion- isants. Chimie. Université Paris Sud - Paris XI, 2009. Français. tel-00447089 HAL Id: tel-00447089 https://tel.archives-ouvertes.fr/tel-00447089 Submitted on 14 Jan 2010 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. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. N° D’ORDRE : 9602 UNIVERSITÉ PARIS SUD XI FACULTÉ DES SCIENCES D’ORSAY THÈSE DE DOCTORAT Présentée en vue de l’obtention du grade de DOCTEUR EN SCIENCES DE L’UNIVERSITÉ PARIS SUD XI Spécialité Chimie Organique Par Brice NADAL Ingénieur CPE Lyon Synthèse et évaluation de nouveaux agents de protection contre les rayonnements ionisants Soutenue le 29 octobre 2009 devant la commission d’examen : Professeur Cyrille Kouklovsky Président Docteur Paul-Henri Ducrot Rapporteur Professeur Olivier Piva Rapporteur Docteur Claude Lion Examinateur Docteur Pierre Bischoff Examinateur Docteur Thierry Le Gall Directeur de thèse N° D’ORDRE : 9602 UNIVERSITÉ PARIS SUD XI FACULTÉ DES SCIENCES D’ORSAY THÈSE DE DOCTORAT Présentée en vue de l’obtention du grade de DOCTEUR EN SCIENCES DE L’UNIVERSITÉ PARIS SUD XI Spécialité Chimie Organique Par Brice NADAL Ingénieur CPE Lyon Synthèse et évaluation de nouveaux agents de protection contre les rayonnements ionisants Soutenue le 29 octobre 2009 devant la commission d’examen : Professeur Cyrille Kouklovsky Président Docteur Paul-Henri Ducrot Rapporteur Professeur Olivier Piva Rapporteur Docteur Claude Lion Examinateur Docteur Pierre Bischoff Examinateur Docteur Thierry Le Gall Directeur de thèse Ce travail de thèse a été financé par la Direction Générale pour l’Armement. Il a été réalisé au CEA de Saclay, dans le Service de Chimie Bioorganique et de Marquage (SCBM) de l’Institut de Biologie et de Technologies de Saclay (iBiTec-S) au sein de la Direction des Sciences du Vivant (DSV). Remerciements Je voudrais en premier lieu remercier le Dr Thierry Le Gall pour la confiance qu'il m'a accordée en m'intégrant dans son équipe. Ses qualités scientifiques et sa passion pour la chimie m'ont permis de mener à bien ce travail de thèse. Je tiens à exprimer ma gratitude au Pr Cyrille Kouklovsky, au Dr Paul-Henri Ducrot, au Pr Olivier Piva, au Dr Claude Lion et au Dr Pierre Bischoff pour avoir accepté de juger ce travail. Mes remerciements s’adressent aussi au Dr Bernard Rousseau, responsable du SCBM, pour m’avoir accueilli au sein de son service mais également pour ses conseils et sa disponibilité. Je tiens à exprimer ma gratitude à tous ceux qui ont contribué au bon déroulement scientifique de ce travail. Je tiens plus particulièrement à remercier le Dr Stéphane Meunier de l'Université de Strasbourg et Marie-Claire Nevers du Service de Pharmacologie et d’Immunologie pour leur patience et leur aide mais également pour m'avoir initié aux dosages immuno-enzymatiques. Je remercie le Dr Serge Pin et le Dr Jean-Philippe Renault du Laboratoire de Radiolyse pour leurs conseils, leurs contributions ainsi que pour nos nombreuses discussions sur les tests biologiques sous irradiations. Un grand merci au Dr Pierre Bischoff, au Dr Damien Habrant, à Antoine Le Roux de l’Université de Strasbourg, au Dr Ghassoub Rima et à Damien Cressier de l’Université de Toulouse pour les tests antioxydants et les études de radioprotection. Je remercie Hélène pour son travail rigoureux, sa persévérance et son courage. Merci à Sophie pour son aide pour la synthèse de composés et pour la réalisation de tests antioxydants. Je tiens à remercier Elisabeth Zekri, Estelle Léonce, David Buisson du service d’analyse pour leur aide au bon déroulement de ce projet. Un très grand merci à Aurélie qui a été d'un grand soutien au labo pendant mes deux premières années de thèse dans les moments difficiles où la chimie ne marchait pas … Et merci Delphine d'avoir pris le relais pendant cette dernière année. Nous avons beaucoup discuté ces derniers mois et ton amitié m'a permis de tenir le coup dans des moments difficiles. Je remercie chaleureusement tous les permanents, thésards, post-docs et stagiaires de la « Recherche » que j’ai rencontrés pendant ces trois années et plus particulièrement Yves, Marie, Xavier, Julien C., Julien R., Fanny, Véro … pour l’ambiance agréable qui régnait dans le laboratoire. Je tiens également à adresser mes remerciements à tous mes collègues des labos tritium, 14 C et de la combi pour leur accueil sympathique et leurs soutiens occasionnels. Je remercie également Chantal et Eric Le Gallo. Je souhaite remercier toutes les personnes d’Eurisotop pour leur bonne humeur et tous les petits services qu’ils peuvent nous rendre : Fredo, Yannick, Géraldine, Fredy ainsi qu’Alain Valleix et tous les autres. Je remercie chaleureusement ma famille sans qui tout cela n'aurait été possible et tout particulièrement mes parents, mes frères et mes grands-parents. Je souhaite également remercier Françoise et Yves pour m'avoir chaleureusement accueilli et soutenu. Enfin, un grand merci à Caro pour m'avoir accompagné et encouragé pendant ces dernières années. Ta présence et ton soutien au quotidien sont d'un grand réconfort et me permettent d'avancer. Abréviations Abréviations A AAPH : dichlorhydrate de 2,2’-azobis-2-méthyl-propanimidamide ABTS : acide 2,2’-azinobis-(3-éthylbenzothiazoline-6-sulfonique) Ac : acétyle AChE : acétylcholinestérase ADN : acide désoxyribonucléique AIBN : azobis(isobutyronitrile) AMT : 3’-amino-3’-déoxythymidine Ar : aryle ARP : pouvoir antiradicalaire B BER : excision de bases BHA : 3-tert -butyl-4-hydroxyanisole Bn : benzyle Boc : tert -butoxycarbonyle B-PE : B-phycoérythrine BSA : albumine sérique bovine Bu : butyle C CAN : nitrate de cérium ammoniacal CAS : anticorps polyclonal de chèvre anti-immunoglobulines de souris CCM : chromatographie sur couche mince CEA : Commissariat à l’Energie Atomique CNRS : Centre National de la Recherche Scientifique CUPRAC : cupric reducing antioxidant capacity D DBN : 1,5-diazabicyclo[4.3.0]non-5-ène DBU : 1,8-diazabicyclo[5.4.0]undéc-7-ène DCC : N,N’-dicyclohexylcarbodiimide DGA : Direction Générale pour l’Armement DL : dose létale DMAP : 4-(diméthylamino)pyridine DMF : N,N-diméthylformamide DMPU : 1,3-diméthyl-3,4,5,6-tétrahydro-2(1 H)-pyrimidinone DMSO : diméthylsulfoxyde DPPH : 2,2-diphényl-1-picrylhydrazyle DTNB : 5,5’-dithiobis(acide 2-nitrobenzoïque) E EAO : espèces activées de l’oxygène ED : dose efficace EDC : 1-éthyl-3-(3-diméthylaminopropyl)carbodiimide EDTA : acide éthylènediaminetétraacétique Abréviations EIA : dosage immuno-enzymatique éq. : équivalent ESI : ionisation par électrospray Et : éthyle F FDA : Food and Drug Administration FRAP : ferric reducing antioxidant power G Gp : groupement protecteur H HMBC : corrélation hétéronucléaire à longues distances HR : réparation par recombinaison homologue HRMS : spectrométrie de masse à haute résolution HSQC : correlation hétéronucléaire à simple quantum I ID : dose infectieuse Ig : immunoglobuline IR : infrarouge IRAMIS : Institut Rayonnement et Matière de Saclay K KMBA : acide 2-céto-4-méthiobutyrique L LDA : diisopropylamidure de lithium LICA : N-cyclohexyl-N-isopropylamidure de lithium LiHMDS : bis(triméthylsilyl)amidure de lithium Litt . : littérature LNS : liaison non spécifique M MDA : malonaldéhyde Me : méthyle Ms : méthanesulfonyle MS : spectrométrie de masse N NBS : N-bromosuccinimide NER : excision de nucléotides NHEJ : réparation par religation non homologue NIS : N-iodosuccinimide O ORAC : Oxygen Radical Absorbance Capacity Abréviations P PBS : Phosphate Buffered Saline Ph : phényle ppm : partie par million PROPERGAL : PROtection du PERsonnel – Nouvelle formulation GALénique R rdt : rendement Rf : rapport frontal RMN : résonance magnétique nucléaire RPE : résonance paramagnétique électronique S SBIGeM : Service de Biologie Intégrative et Génétique Moléculaire SCBM : Service de Chimie Bioorganique et de Marquage SIS2M : Service Interdiciplinaire sur les Systèmes Moléculaires et les Matériaux SOD : superoxyde dismutase SPI : Service de Pharmacologie et d’Immunoanalyse T t.a. : température ambiante TBA : acide thiobarbiturique TBAF : fluorure de tétrabutylammonium t-Bu : tert -butyle Tf : triflate TFA : acide trifluoroacétique TFAA : anhydride trifluoroacétique THF : tétrahydrofurane TMS : triméthylsilyle TOF : temps de vol TOSC : Total Oxyradical Scavenging Capacity TRAP : Total Radical trapping Antioxidant Parameter Ts : tosyle U UAF : unité arbitraire de fluorescence UV : ultraviolet Sommaire Sommaire Introduction générale............................................................................................................................ 1 Chapitre 1 : Les rayonnements ionisants : effet et protection........................................................... 5 I. Les rayonnements ionisants........................................................................................................ 5 1. Définition..........................................................................................................................................
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