Latifou LAGNIKA ETUDE PHYTOCHIMIQUE ET ACTIVITE

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Latifou LAGNIKA ETUDE PHYTOCHIMIQUE ET ACTIVITE UNIVERSITE LOUIS PASTEUR STRASBOURG FACULTE DE PHARMACIE en cotutelle avec L’UNIVERSITE D’ABOMEY-CALAVI BENIN FACULTE DES SCIENCES ET TECHNIQUES THESE Soutenue à la Faculté de Pharmacie de Strasbourg le 14 décembre 2005 pour obtenir le grade de DOCTEUR de L’ UNIVERSITE LOUIS PASTEUR (Mention Sciences Pharmaceutiques) Domaine : PHARMACOGNOSIE présentée par Latifou LAGNIKA ETUDE PHYTOCHIMIQUE ET ACTIVITE BIOLOGIQUE DE SUBSTANCES NATURELLES ISOLEES DE PLANTES BENINOISES JURY Monsieur B. WENIGER Directeur de Thèse Monsieur le Professeur A. SANNI Co-Directeur de thèse Monsieur le Professeur M. HIBERT Rapporteur interne Madame le Professeur V. COLLOT Rapporteur externe Monsieur le Professeur K. DRAMANE Rapporteur externe non membre du jury Nous tenons à remercier : Monsieur Bernard WENIGER, Directeur de thèse Maître de conférences en Pharmacochimie des Substances Naturelles à l’Université Louis Pasteur de Strasbourg & Monsieur Ambaliou SANNI, co-Directeur de thèse Professeur de Biochimie et biologie moléculaire à la Faculté des Sciences et Techniques de l’Université d’Abomey- calavi, Bénin qui, durant ces trois années, ont bien voulu diriger mes travaux de thèse, en me faisant bénéficier de leur expérience, leur compétences et encouragements. Monsieur Marcel HIBERT Professeur de Pharmacochimie à la Faculté de Pharmacie de l’Université de Strasbourg Madame Valérie COLLOT Professeur de Pharmacognosie à l’Université de Caen qui nous ont fait l’honneur d’accepter de juger et de siéger dans le jury de thèse. En témoignage de ma reconnaissance et respectueuse gratitude Nous tenons à remercier : L’Agence Universitaire Francophone (AUF), dans le cadre du projet scientifique inter- universitaire intitulé : Activité antipaludique et action réversante de la chloroquinorésistance d’extraits d’espèces médicinales et de biomolécules purifiées issues des pharmacopées traditionnelles béninoise et ivoirienne. L’Ambassade de France au Bénin qui nous a attribué une bourse en alternance. L’Union International de Biochimie et Biologie Moléculaire (IUBMB) pour l’allocation qu’elle nous a attribuée. Nos remerciements vont également à toute les personnes qui ont contribué de près ou de loin à la réalisation de ce travail, et notamment à : Madame Catherine VONTHRON-SENECHAU Pour son aide constante et ses précieux conseils pendant tout la durée de ce travail. J’ai été très content d’avoir travaillé avec toi. Merci Monsieur le Professeur Robert ANTON Directeur du Laboratoire de Pharmacognosie à la Faculté de Pharmacie de Strasbourg Pour l'accueil chaleureux que vous nous avez toujours réservé dans votre Laboratoire et je souhaite que cette collaboration initiée se poursuive. Veuillez trouver ici l'expression de notre profonde gratitude. Madame Annelise LOBSTEIN Pour votre accueil dès notre arrivé au Laboratoire, vos conseils et votre disponibilité. Mes sincères remerciements. Monsieur Bernard KUBALLA Pour votre simplicité et disponibilité Martine BERNARD Pour sa bonne humeur et disponibilité. Sincères remerciements Monsieur le Professeur Jacques Henry WEIL Pour votre engagement, votre aide et votre soutien quotidien durant toutes ces années. Vous êtes l’un des pionniers de la création des licence et maîtrise de Biochimie au Bénin. Toute notre gratitude. Monsieur Cyril ANTHEAUME Pour sa gentillesse, sa grande compétence dans le domaine de la RMN et sa contribution à l’élucidation structurale des composés. Monsieur le Professeur Karim DRAMANE Rapporteur externe : vous avez accepté, prendre de votre temps, pour juger cette thèse. Tous nos remerciements. Monsieur le Professeur Mansour MOUDACHIROU Pour votre disponibilité et votre encouragement. Mes sincères remerciements. Docteur Lamine Saïd BABA MOUSSA, Madame Lucie AYI FANOU, Monsieur Nicodème CHABI. Pour vos conseils et encouragement pendant les moments les plus difficiles. Nous ne saurions oublier tous nos collègues et stagiaires du Laboratoire de Pharmacognosie et du Laboratoire de biochimie et Biologie moléculaire de l’UAC : Saliou N’GOM, CHAABI Mehdi, Inès EDAYE, Tom ADJOBIMEY, Patrice AVOGBE, Bérénice ASSOGBA et Michael MISSIHOUN, avec qui nous avons toujours su entretenir une ambiance chaleureuse et amicale. ABREVIATIONS AcOEt : Acétate d’éthyle ADN : Acide Désoxyribonucléique ALCl3 : Chlorure d’aluminium APCI : Ionisation Chimique à Pression Athmosphérique CCM : Chromatographie sur Couche Mince CH3CN (ACN) : Acétonitrile DDT : Dichloro-diphenyl-trichloroéthane DHEA : Déhydroépiandrostérone DHFR : Dihydrofolate synthétase DHPS : Dihydroptéroate synthétase DMSO : Diméthylsulfoxyde EDTA : Acide ethylenediaminetetraacétique ELISA : Enzyme-Linked Immunosorbent Assay ESI : Electrospray (ionisation par électronébulisation) EtOH : Ethanol FAB : Fast Atom Bombardment FBS B : Fast Blue Salt B (3,3’-diméthoxy-biphenyl-4,4’-bis (diazonium dichloride)) HCH : Hexachlorocyclohexane HGPRT : Hypoxanthine-guanine phosphoribosyle transférase HMBC : Heteronuclear Multiple Bond Correlation HRP : Histidine Rich Protéine HSQC : heteronuclear single quantum correlation IC50 : Concentration Inhibitrice 50 IE : Impact Electronique IgG : Immunoglobuline G IgM : Immunoglobuline M Jmod : J-modulated spin-echo LDH : Lactate déshydrogénase NAD : Nicotinamide adénine dinucléotide NaHCO3 : Carbonate de sodium NaOH: Hydroxyde de sodium NH4OH: Hydroxyde d’ammonium NP/PEG : Natural Product Polyethylèneglycol NOESY : Nuclear overhauser effect spectroscopy OMS : Organisation Mondiale de le Santé PMA : phorbol myristate acetate PCR : Polymérase Chain Reaction QBC : Quantitative Buffy Coat RMN : Résonance Magnétique Nucléaire RNA : Acide ribonucléique Rf : Rapport Frontal RPMI : Roswell Park Memorial Institute Medium SbCl : Chlorure d’antimoine SNIGS : Système national d'Information et de Gestion Sanitaire TOCSY : Total Correlated Spectroscopy WHO: World Health Organization Sommaire INTRODUCTION……………………………………………………………………. 1 Première partie : ETUDE BIBLIOGRAPHIQUE…………………………………. 6 Chapitre 1 : GENERALITES SUR LE PALUDISME…………………………….. 7 1. Epidémiologie du paludisme………………………………………………….. 8 1.1 Répartition géographique du paludisme………………………………… 8 1.2 Des chiffres alarmants………………………………………………...… 9 2. Le parasite et le vecteur…………………………………………...................... 11 2.1 Le parasite du genre«Plasmodium»………………………..………...…. 11 2.2 Le vecteur «l’Anophèle femelle»…………………………………...…... 12 2.3 Cycle de reproduction des plasmodies……………...…………………... 13 2.3.1 Cycle asexué ou schizogonique chez l’homme…………………………. 13 2.3.2 Cycle sexué ou sporogonie chez l’Anophèle…………………………… 14 3. Symptomatologie………………………………………………………………. 16 3.1 Accès de primo-invasion………………………………………………... 16 3.2 L’invasion……………………………………………………………….. 16 3.3 Les formes cliniques…………………………………………………….. 17 3.4 Les reviviscences………………………………………………………... 17 3.5 Le rythme des accès…………………………………………………….. 18 3.6 Particularités symptomatiques liées aux espèces……………………….. 18 3.7 Fièvre bilieuse hémoglobinurique………………………………………. 19 4. Diagnostic………………………………………………………………………. 20 4.1 Diagnostic clinique……………………………………………………… 21 4.2 Diagnostic basé sur la mise en évidence du parasite……………………. 22 4.2.1 Diagnostic microscopique classique……………………………………. 22 4.2.2 Diagnostic basé sur la détection d’antigènes parasitaires………………. 23 4.2.3 Diagnostic basé sur la détection du matériel génomique du parasite…… 24 5. Traitement du paludisme……………………………………………………… 25 5.1 Les produits disponibles………………………………………………… 26 5.1.1 Les schizontocides………………………………………………………. 26 5.1.2 Les gamétocytocides………...………………………………………….. 29 5.1.3 Les antibiotiques………………………………………………………... 29 5.1.4 Les associations d’antipaludiques………………………………………. 29 5.2 Les produits d’avenir……………………………………………………. 30 5.3 Traitement du paludisme par la médecine traditionnelle …………….… 30 6. Prévention……………………………………………………………………… 31 6.1. Prophylaxie générale……………………………………………………. 32 6.2. Prophylaxie individuelle……………………………………………...… 33 Chapitre 2 : NOUVELLES SUBSTANCES ANTIPALUDIQUES ET LES CIBLES THERAPEUTIQUES…………………..…………………… 34 1. Quelques substances naturelles antipaludiques……………………………... 36 2. Cibles chimiothérapeutiques potentielles chez le Plasmodium……………… 41 2.1. Cibles responsables des processus se produisant dans la vacuole digestive du Plasmodium……………………………………………….. 41 2.1.1. La dégradation de l’hémoglobine……………………………………….. 42 2.1.2. La polymérisation de l’hème……………………………………………. 42 2.1.3. Le stress oxydatif……………………………………………………….. 43 2.2. Cibles intervenant dans la production des enzymes impliquées dans la synthèse macromoléculaires et des métabolites……………………… 44 2.2.1. La glycolyse…………………………………………………………….. 44 2.2.2. Le métabolisme des acides nucléiques………………………………….. 44 Chapitre 3: DESCRIPTION DES PLANTES ETUDIEES……………………...… 48 1. La famille des Amaranthaceae et l’espèce Gomphrena celosioides…………. 50 1.1 La famille des Amaranthaceae………………………………………….. 51 1.1.1. Présentation……………………………………………………………... 51 1.1.2 Intérêt pharmacologique, nutritionnel et commercial…………………... 51 1.1.3 Chimie des Amaranthaceae……………………………………………... 52 1.2 Le genre Gomphrena……………………………………………………. 53 1.2.1 Présentation……………………………………………………………... 53 1.2.2 Propriétés pharmacologiques des Gomphrena…..……………………… 54 1.2.3 Quelques exemples de composés isolés du genre Gomphrena……...….. 55 1.3 L’espèce Gomphrena celosioïdes……………………………………….. 57 1.3.1 Classification……………………………………………………………. 59 1.3.2 Description botanique…………………………………………………... 59 1.3.3 Utilisation dans la médecine traditionnelle……………………………... 60 2. La famille des Euphorbiaceae et l’espèce Croton lobatus………….………… 61 2.1 La famille
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