Phytochimie-Chimie Organique Biologique

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Phytochimie-Chimie Organique Biologique N° d’ordre : 2015AGUY0858 Faculté des Sciences Exactes et Naturelles de Guadeloupe Ecole doctorale pluridisciplinaire THESE En vue de l’obtention du DOCTORAT Discipline : chimie Spécialité : phytochimie-chimie organique biologique Présentée par Josianny JOSEPH-ANGELIQUE CONTRIBUTION A LA CONNAISSANCE CHIMIQUE ET VALORISATION BIOLOGIQUE DE Nectandra membranacea (Swartz) Grisebach de Guadeloupe Soutenue le 23 juillet 2015 COMPOSITION DU JURY Pr André PICHETTE Laboratoire d’Analyse et de Séparation Rapporteur des Essences VEgétales (LASEVE) Université du Québec à Chicoutimi Pr Laila SALMEN Laboratorie de Farmacognosia Rapporteur ESPINDOLA DARVENNE Universidade de Brasília Pr Jacqueline ABAUL Université des Antilles et de la Guyane Examinateur Pr Joëlle LEVALOIS Université des Antilles et de la Guyane Directeur de GRÜTZMACHER thèse ii REMERCIEMENTS Ce travail a été effectué à l'Unité Mixte de Recherche de la Qualité et Valorisation des produits végétaux tropicaux (UMR QUALITROP 1270) de l'Université des Antilles et de la Guyane, Pôle Guadeloupe, sous la direction de Madame Le Professeur Joëlle LEVALOIS- GRÜTZMACHER. Je tiens à la remercier pour m'avoir accueillie au sein du laboratoire et de m’avoir donné l’opportunité de poursuivre ma formation en phytochimie. Je tiens également à remercier Madame Sylvie BERCION, directrice du laboratoire QualiTrop et Maître de conférence-HDR en Chimie, pour ses conseils, son soutien et ses encouragements. Je remercie également mes responsables de thèse, à l’origine du projet sur lequel est basé cette thèse, Madame Lyn UDINO et Madame Muriel SYLVESTRE, Maîtres de conférences en Chimie, pour leurs efforts, leurs conseils et leur positivité, tout au long de ces années de travail. J'aimerais également exprimer ma gratitude à Madame Le Professeur Jacqueline ABAUL, Directrice de l'Ecole Doctorale et Professeur en Chimie, pour ses encouragements et sa compréhension. Je remercie Monsieur JOVIAL, de m'avoir permis de récolter sur son terrain, l'objet de mon étude, sans lui ce travail n'aurait pas pu être réalisé. Mes remerciements vont également aux rapporteurs les Professeurs Laila SALMEN ESPINDOLA DARVENNE et André PICHETTE qui ont accepté de juger ce travail. Je remercie également les personnes dont la collaboration a été essentielle pour plusieurs aspects de ce travail : Les Professeurs André PICHETTE et Jean LEGAULT, de l’Université du Québec à Chicoutimi et Directeurs du Laboratoire d’Analyse et de Séparation des Essences Végétales (LASEVE) de m’avoir accueillie et permis de réaliser sur leur site diverses analyses et tests biologiques sur mes échantillons. iii Monsieur Le Professeur, Hanszork GRÜTZMACHER m'a accueillie dans son laboratoire à l’Ecole polytechnique fédérale de Zurich, ETH. Je remercie toute son équipe pour leur convivialité et leur gentillesse. J'y ai passé de très bons moments. Monsieur Le Professeur Etienne HIRSCH, neurobiologiste, et Directeur de recherche au CNRS (Centre national de la recherche scientifique) et à l’INSERM (Institut National de La Santé et de la Recherche Médicale) qui a réalisé les tests neuroprotecteurs pour la maladie d'Alzheimer sur mes extraits. Je suis aussi très reconnaissante envers Monsieur Serge LAVOIE pour avoir partagé avec moi ses connaissances et pour sa disponibilité. Ce travail n'aurait pas pu être mené à bien sans une équipe de collègues : Une mention particulière pour Madame Francine LAVOIE-NAGAU qui m’a énormément soutenue, conseillée, aidée dans ce travail et merci pour sa bonne humeur au quotidien. Merci beaucoup Francine! Je remercie tous mes collègues de bureau, particulièrement Grégory LEGRAVE pour m'avoir encouragée, pour sa disponibilité et pour m'avoir supportée durant cette thèse. Corine, Laurent, Axelle, Lisa et Sabin pour tout leur soutien, leurs conseils et leur bonne humeur. Merci à ma famille de cœur, Madame Novelta MALINUR dit Tata, Monsieur Serge Malinur, à Denise, Marie-Line, Fulbert, Stacy, Shannie, Allan et Nicolas pour leur soutien et les bons moments passés ensemble. Un merci particulier pour Jocelyn et François pour m'avoir aidée lors de mes nombreuses récoltes, et pour leurs encouragements. Je remercie également toute l'équipe de l'UFR Sciences Exactes et Naturelles. Je tiens à remercier aussi Rod DABRIOU, Harry MEPHON pour leur soutien ainsi que Rio, Constant, Véronique, Jean-Jacques, Daniel et tout le personnel du centre hospitalier du Carbet. Enfin mes remerciements vont aussi à Roberto, Lulu, Julien, Jean-Claude et Norbert pour m’avoir soutenue moralement, encouragée à surmonter ces épreuves difficiles. Et également au Dr Carra pour avoir contribué à la présentation de ce travail. A toute ma famille, mes parents, Alex BENGON dit Lailai et tous mes amis Didier, Christelle, Constance et Eve, merci. iv Résumé Contribution à la connaissance chimique et à la valorisation biologique de Nectandra membranacea (Swartz) Grisebach Nectandra membranacea (Lauraceae) est une plante très commune aux Petites Antilles. Aucune utilisation traditionnelle de cette plante n’est connue mais le genre Nectandra, qui représente le deuxième plus important de la famille des Lauraceae avec 114 espèces reconnues, présente des propriétés analgésiques, anti-inflammatoires et énergétiques (Le Quesne et al 1980). Nectandra a également démontré une activité antitumorale (Silva-Filho, 2004). Les nombreuses propriétés du genre et le peu de connaissance chimique sur cette espèce nous ont incités à réaliser une étude phytochimique et à évaluer le potentiel biologique des différentes parties de cette plante. L’analyse des lipides a été effectuée, et les principales classes de métabolites secondaires (terpènes, flavonoïdes, alcaloïdes) ont été recherchées dans les feuilles, les écorces et les fruits de Nectandra membranacea. Afin de suivre les variations de la composition chimique en fonction du cycle de vie du végétal, l’étude a été réalisée sur des échantillons (feuilles, écorces et fruits), récoltés à deux stades du cycle de la plante : la germination (stade A) et la fructification (stade B). N. membranacea, grâce à la présence de nombreuses familles chimiques, possède des activités biologiques intéressantes. Les huiles essentielles ont montré des activités anticancéreuses, anti-inflammatoires, antioxydantes et antiherpétiques. Les extraits polyphénoliques issus des différentes parties de la plante ont montré une très grande activité antioxydante (ORAC et DPPH) et in vitro (cellules saines) ainsi que des propriétés anti-inflammatoires notables. Les extraits d’alcaloïdes totaux issus des trois parties ont été évalués pour la maladie d’Alzheimer et ont révélé de très bons effets neuroprotecteurs, plus particulièrement pour les extraits de feuilles récoltées lors de la germination. Ce travail a permis d’élargir la connaissance des huiles essentielles de l’espèce (composition pour plusieurs parties et activité biologique). Des flavonoïdes ont été identifiés, mais la plante s’est révélée particulièrement riche en alcaloïdes (une quinzaine) de type isoquinoliques et indoliques. N. membranacea se révèle être une plante aux nombreuses propriétés, au regard de ses activités biologiques intéressantes. Des extraits pourraient être utilisés comme principes actifs dans des médicaments mais également en industrie cosmétique. Mots clés : Nectandra membranacea, Lauraceae, étude phytochimique, activité biologique, cytotoxique, anti-inflammatoire, antioxydant, antiviral. v Abstract Contribution to chemical knowledge and biological recovery of Nectandra membranacea (Swartz) Grisebach Nectandra membranacea (Lauraceae) is a very common plant in the French West Indies. No traditional use of this plant is known, but the genus Nectandra, which represents the second most important of Lauraceae family with 114 recognized species, has got analgesic, anti-inflammatory and energizing activities (Le Quesne et al 1980). Nectandra was also identified as a potential antitumor agent (Silva-Filho, 2004). Many properties of the genus, and some chemical knowledge of this species, have prompted us to perform phytochemical and biological study of different parts of the plant. Lipids and the main classes of secondary metabolites (terpenes, flavonoids, alkaloids) were investigated in leaves, bark and fruits of Nectandra membranacea. To determine the monitoring changes in the chemical composition, according to the life cycle of the plant, the study was carried out on samples (leaves, bark and fruits) harvested at two stages of the cycle plant : germination (stage A) and fruit stage (stage B). N. membranacea, thanks to the presence of many chemical families, has interesting biological activities. Essential oils have shown anticancer, anti-inflammatory, antioxidant activities. They revealed antiviral activity on the herpes virus. The polyphenolic extracts from different parts of the plant showed a high antioxidant activity (ORAC and DPPH tests) and in vitro (healthy cells) and significant anti-inflammatory properties. The total alkaloids extracted from the three parts were evaluated for Alzheimer’s disease and were found to have very good protective effects, especially the leaves’ extracts collected during germination. This work has expanded knowledge of the essential oil of the species (composition of several parts and biological activity). Flavonoids have been identified, but the plant was shown to be particularly rich in alkaloids (fifteen were identified) of indolic and isoquinolic type. N. membranacea presents many properties. In view of its interesting biological activities, it could
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