A presente dissertação descreve um estudo químico das cascas do tronco de Guarea guidonia (L.) Sleumer (Meliaceae). Do extrato hexânico de cascas coletadas de um espécimen localizado em Campo Grande (MS) foram isolados e identificados cinco sesquiterpenos: o trans 1(10)-epóxi-4(15)-cariofileno; o 1(10)-epóxi-4,7-humuladieno; o 10-aloaromadendranol (viridiflorol); o 1(10),4-diepóxi-7-humuleno e o 3-oxa-10-aloaromadendranol e quatro esteróides: o sitosterol, o estigmasterol, o palmitato do sitosterol e o oleato do sitosterol. Esteróides glicosilados foram também detectados nesse extrato. Uma destilação por arraste a vapor d\'água forneceu o óleo essencial que foi comparado por cromatografia gasosa, com o óleo extraído de cascas de uma planta localizada em Araraquara (SP). Os cromatogramas dos dois óleos indicam que eles possuem composições químicas muito semelhantes. Do óleo essencial obtido do espécimen de Araraquara foram identificados onze sesquiterpenos: o 1(10),4(15),5-germacratrieno (germacreno D), o cis 4,10(14)-cadinadieno (γ-muuroleno), o trans 4,10(14)-cadinadieno, o 1(10),4-cadinadieno (σ-cadineno), o 1(10),4(15)-cariofiladieno, o 1,3,11-elematrieno (β-elemeno), o 1(10),4,7-humulatrieno (α-humuleno), o cis e o trans 1(10)-epóxi-4(15)-cariofileno, o 1(10)-epóxi-4,7-humuladieno e o 10-aloaromadendranol (viridiflorol). Os três últimos sesquiterpenos foram encontrados também no extrato hexânico do espécimen que vive em Campo Grande. As substâncias foram isoladas através de cromatografias de adsorção em sílica gel e desta impregnada com AgNO3 e identificadas por espectroscopias de RMN de 1H e de 13C (PND e DEPT 1350) e de massas. / This work describes a chemical study of the bark wood of Guarea guidonia (L.) Sleumer (Meliaceae). The hexanic extract of barks collected from an especimen growing at Campo Grande (MS) afforded five sesquiterpenes: the trans 1(10)-epoxi-4(15)-cariofileno; the 1(1 0)-epoxi-4,7-humuladieno; the 10-aloaromadendranol (viridiflorol); the 1(10),4-diepoxi-7-humuleno and the 3-oxa-10-aloaromadendranol and four steroids: the sitosterol, the stigmasterol, the palmitato of sitosterol and the oleato of sitosterol. From this extract were also detected glycosyl steroids. The essencial oil obtained from a steam water extraction of this barks was compared with that extract fiom the barks of an Araraquara\'s plant (SP). The chromatograms of the oils showed that the chemical composition of both oils are very similar. From the Araraquara\'s oil were identified eleven sesquiterpenes: the 1(10),4(15),5-germacratrieno (germacreno D), the cis 4,10(14)-cadinadieno (γ-muuroleno), the trans 4,10(14)-cadinadieno, the 1(10),4-cadinadieno (σ-cadineno), the 1(10),4(15)-cariofiladieno, the 1,3,11-elematrieno (β-elemeno), the 1(10),4,7-humulatrieno (α-humuleno), the cis and the trans 1(10)-epoxi-4(15)-cariofileno, the 1(10)-epoxi-4,7-humuladieno and the 10-aloaromadendranol (viridiflorol). The last three sesquiterpenes were also isolated fiom the hexanic extract. The compounds were isolated by adsortion chromatography on silica gel and on silica gel impregnated by AgNO3 and identified by 1H and 13C NMR (PND and DEPT 1350) and mass spectroscopy.

This thesis investigated the phytochemistry and pharmacology of immunomodulatory medicinal plant species used traditionally by the Q’eqchi’ Maya Healers Association (QMHA) of Belize, and the Cree of Eeyou Istchee (CEI) of northern Quebec. Using quantitative ethnobotanical methodology, we identified 107 plant species belonging to 49 families used by Q’eqchi’ healers in the treatment of symptoms from 14 usage categories related to inflammation. Regression analysis revealed that the Piperaceae, Araceae, and Begoniaceae are preferentially selected by the Maya. Healer consensus for plant species was high, with 56 species (52%) being used by all the healers, and consensus for usage categories was also high, as informant consensus factor (FIC) values for each category were greater than 0.4. Fifty-two Belizean species were evaluated for their TNF-α inhibitory activity in an LPS-stimulated THP-1 monocyte model. Twenty-one species (40%) demonstrated significant TNF-α inhibition when assayed at 100 µg/mL, 8 of which had greater than 50% of the activity of the parthenolide positive control (10 µg/mL). Significant regressions were found between the anti-inflammatory activity and total healer frequency of use (Fuse) and the use reports for 3 usage categories, which indicated that ethnobotanical parameters can in part predict the activity of traditionally used species. Five sesquiterpene lactones were isolated from the leaves of Neurolaena lobata, one of the most active species tested, all of which demonstrated anti-inflammatory activity greater than that of parthenolide (IC50 = 4.79 µM), with IC50s ranging from 0.17-2.32 µM. Lobatin B was the most active isolate tested. Ethanolic and water extracts of 17 species used by Cree healers were evaluated for their immunomodulatory activity. In general, the average anti-inflammatory activity of ethanolic extracts was 1.8 times greater than that of water extracts, and the pro-inflammatory activity of water extracts was 3.7 times greater than ethanolic extracts. Picea mariana and Pinus banksiana were the most anti-inflammatory ethanolic and water extracts, while the water extract of Sarracenia purpurea was the most pro-inflammatory. Picea marina cones, the most anti-inflammatory Cree medicine, were subjected to bioassay guided isolation. This led to the isolation of the anti-inflammatory lignan (+)-lariciresinol-9’-p-coumarate, which had an IC50 of 28.4 µM. Together, these results validate the traditional knowledge shared by our Q’eqchi’ and Cree collaborators, and draw attention to the therapeutic potential of subtropical and boreal plants as culturally appropriate complements to modern medicine. - Cette thèse porte sur la phytochimie et la pharmacologie des espèces de plantes médicinales immunomodulatrices utilisées traditionnellement par le Q’eqchi’ Maya Healers Association (QMHA) du Belize, et les Cris d'Eeyou Istchee (CEI) du nord du Québec. En utilisant une méthodologie ethnobotanique quantitative, nous avons identifié 107 espèces végétales appartenant à 49 familles utilisées par les guérisseurs Q'eqchi’ dans le traitement de symptômes appartenant à 14 catégories d'utilisation liées à l'inflammation. Une analyse de régression a révélé que les familles Piperaceae, Araceae, et Begoniaceae sont préférentiellement choisis par les Mayas. Le consensus entre guérisseurs pour les espèces végétales était élevé, avec 56 espèces (52%) étant utilisés par tous les guérisseurs, et le consensus pour les catégories d'utilisation était également élevé, car les valeurs de facteur de consensus des informants (FIC) pour chaque catégorie étaient supérieurs à 0,4. Cinquante-deux espèces du Belize ont été évaluées pour leur activité inhibitrice de TNF-α dans un modèle de THP-1 monocytes stimulés par le LPS. Vingt-et-une espèces (40%) ont montré une inhibition significative de TNF-α lorsque dosés à 100 µg/mL, dont 8 d’entre elles ont démontrées plus de 50% de l'activité du contrôle positif parthénolide (10 µg/mL). Des régressions significatives ont été observées entre l'activité anti-inflammatoire et la fréquence d'utilisation de guérisseurs totale (Fuse) et les rapports d'utilisation pour 3 catégories d'utilisation, ce qui indique que les paramètres ethnobotaniques peuvent en partie prédire l'activité des espèces traditionnellement utilisées. Cinq lactones sesquiterpéniques ont été isolés à partir des feuilles de Neurolaena lobata, l'une des espèces les plus actives testées, qui a démontré une activité anti-inflammatoire supérieure à celle du parthénolide (CI50 = 4,79 µM), avec des CI50 allant de 0,17 à 2,32 µM. Lobatin B était l’isolât le plus actif testé. Des extraits éthanoliques et aqueux de 17 espèces utilisées par les guérisseurs Cris ont été évalués pour leur activité immunomodulatrice. En général, l'activité anti-inflammatoire moyenne des extraits éthanoliques était 1,8 fois supérieure à celle des extraits d'eau, et l'activité pro-inflammatoire des extraits d'eau était de 3,7 fois supérieure à celle des extraits éthanoliques. Picea mariana et Pinus banksiana étaient les extraits éthanoliques et aqueux avec le plus d’activité anti-inflammatoire, tandis que l'extrait aqueux de Sarracenia purpurea était le plus pro-inflammatoire. Le cône de Picea marina, le médicament traditionnelle Cris le plus anti-inflammatoire, a été soumis à une isolation guidée par essais biologiques. Cela a mené à l'isolement du lignane anti-inflammatoire (+)-lariciresinol-9'-p-coumarate, qui avait une CI50 de 28,4 µM. Ensemble, ces résultats valident les connaissances traditionnelles partagées par nos collaborateurs Q'eqchi' et Cris, et mettent en évidence le potentiel thérapeutique des plantes subtropicales et boréales comme des compléments à la médecine moderne qui sont culturellement appropriées.

Ce travail de thèse s'inscrit dans un projet régional, initié par le Parc du Lubéron et en collaboration avec le Laboratoire de Pharmacognosie et d'Ethnopharmacologie de l’Université de Marseille. Il avait pour objectif d'étudier la possibilité de domestiquer une plante sauvage, Inula montana L. (Asteraceae) connue dans la pharmacopée provençale pour ses effets anti traumatiques semblables à ceux d'Arnica montana L. et de la proposer comme nouvelle production agricole. Inula montana produit notamment des lactones sesquiterpènes, identifiées comme les métabolites secondaires responsables de ses aptitudes biologiques anti inflammatoires. Il s’est agit dans cette étude1) de déterminer les caractéristiques de croissance et de développement de la plante sauvage largement inconnue et d’identifier en conditions naturelles les facteurs les plus favorables à la production de métabolites secondaires, 2)d’étudier sa capacité à se multiplier végétativement in vitro et à former des cultures cellulaires aptes à synthétiser les molécules d’intérêt 3), de proposer un itinéraire technique applicable agronomiquement et 4) de tester les effets de divers facteurs environnementaux (fertilisation, apport de NaCl, modification du rythme circadien de l’éclairement,rayonnement UVB, ablation de feuilles, application de méthyl jasmonate ) sur la production qualitative et quantitative des lactones sesquiterpènes et des composés phénoliques. Les traits phénologiques de la plante sauvage sont impactés par l’altitude qui induit un retard dans la croissance végétative et la phase reproductrice ainsi que des modifications physiologiques et morphologiques. Les teneurs en métabolites secondaires (certaines lactones sesquiterpènes, les polyphénols totaux et flavonoïdes) varient en fonction de la saison et sont plus importantes dans le site qui présente les conditions climatiques les plus contraignantes du point de vue hydrique (sol drainant,température plus élevée et présence d’une période sèche en été). L’observation microscopique a indiqué la présence de deux types de trichomes : glandulaires (bisériés) et non glandulaires (des poils) qui sont potentiellement les structures porteuses des molécules d’intérêt. I. montana est apte à former des cals in vitro à partir d’explants racinaires, foliaires et caulinaires sur lesquels des pousses feuillées se forment. La domestication d’Inula a été réussie à partir de semences issues des plantes sauvages et en conditions agronomiques, les teneurs en lactones sesquiterpènes (costunolide, artémorine, eldarine et hydrocostunolide) et en composés phénoliques sont généralement plus élevées que chez les plantes sauvages. Les différentes contraintes appliquées pour tester les effets des facteurs environnementaux sur la production des métabolites ont montré : 1) qu’il ne peut pas être établi de corrélation entre la présence d’un stress oxydatif et une augmentation des teneurs en métabolites chez Inula 2) que l’accumulation des lactones et composés phénoliques semblent principalement favorisée lorsque la plante dispose d’un surplus de squelettes carbonés, non utilisés pour la croissance 3) enfin, les deux conditions les plus favorables à l’accumulation des métabolites chez Inula, sont : dans les feuilles, une alternance rapide de lumière et d’obscurité durant la photopériode et dans les capitules, l’application de méthyl jasmonate. Ce travail augure de bonnes perspectives en termes de valorisation d’Inula dans le secteur pharmaco-cosmétologique. Il reste à poursuivre la description du profil phytochimique de la plante et à localiser précisément les organes et/ou sous structures anatomiques concentrant les composés considérés. Ayant démontré que cette plante présente une bonne réponse à la domestication, il est également proposé de poursuivre l’étude des leviers environnementaux susceptibles d’influencer positivement et significativement le profil chimique d’Inula. / This thesis work is part of a regional project, initiated by the Luberon Park and in collaboration with the Laboratory of Pharmacognosy and Ethnopharmacology of the University of Marseille. It aimed to study the possibility of domesticating a wild plant, Inula montana L. (Asteraceae) known in the Provençal pharmacopoeia for its anti-traumatic effects similar to those of Arnica montana L. and to propose it as a new agricultural production. Inula montana produces lactones sesquiterpenes, identified as the secondary metabolites responsible for its biological anti-inflammatory properties. In this study, 1) to determine the growth and development characteristics of the largely unknown wild plant and to identify under natural conditions the most favorable factors for the production of secondary metabolites, 2) to study its characteristics. ability to multiply vegetatively in vitro and to form cell cultures able to synthesize the molecules of interest 3), to propose an agronomically applicable technical itinerary and 4) to test the effects of various environmental factors (fertilization, NaCl supply, modification circadian rhythm of illumination, UVB radiation, ablation of leaves, application of methyl jasmonate) on the qualitative and quantitative production of sesquiterpene lactones and phenolic compounds. The phenological characteristics of the wild plant are impacted by the altitude which induces a delay in the vegetative growth and the reproductive phase as well as physiological and morphological modifications. Levels of secondary metabolites (certain sesquiterpene lactones, total polyphenols and flavonoids) vary according to the season and are more important in the site which has the most water-constraining climatic conditions (draining soil, higher temperature and presence of a dry period in summer). Microscopic observation indicated the presence of two types of trichomes: glandular (biseriate) and non-glandular (hair) which are potentially the carrying structures of the molecules of interest. I. montana is able to form calli in vitro from root, foliar and shoot explants on which leafy shoots are formed. The domestication of Inula has been successful from seed from wild plants and under agronomic conditions, sesquiterpene lactone (costunolide, artemorine, eldarin and hydrocostunolide) and phenolic compounds are generally higher than in wild plants. The different constraints applied to test the effects of environmental factors on the production of metabolites have shown: 1) that there can be no correlation between the presence of oxidative stress and an increase in metabolite levels in Inula 2) that the accumulation of lactones and phenolic compounds seems mainly favored when the plant has a surplus of carbon skeletons, not used for growth; 3) finally, the two most favorable conditions for the accumulation of metabolites in Inula, are: in the leaves, a rapid alternation of light and darkness during the photoperiod and in the flower heads, the application of methyl jasmonate. This work augurs good prospects in terms of valuation of Inula in the pharmaco-cosmetological sector. It remains to continue the description of the phytochemical profile of the plant and to precisely locate the organs and / or anatomical substructures concentrating the compounds in question. Having demonstrated that this plant has a good response to domestication, it is also proposed to continue the study of environmental levers likely to positively and significantly influence the chemical profile of Inula.

For the first time in human history, more people worldwide suffer from obesity than are undernourished. Numerous health complications are associated with obesity including cardiovascular disease, Type 2 Diabetes, cancers of reproductive tissues, stroke, depression, anxiety disorders, and Alzheimer’s disease. A deeper understanding of the anti-adipogenic effects and mechanism of action of sesquiterpene lactones may have pharmacological import in the continuing search for therapeutic modalities to ameliorate the effects of this global obesity epidemic. Dehydroleucodine (DhL), 11,13-dihydro-dehydroleucodine (DH-DhL), and dehydroparashin-B (DhP), sesquiterpene lactones extracted from or derived from compounds extracted from Artemisia douglasiana, were investigated for their anti-adipogenic effects on 3T1-L1 preadipocytes. Dehydroleucodine inhibited the expression of C/EBPa and PPARg, and also strongly blocked the expression of C/EBPβ, an early stage biomarker of early adipogenesis, in a concentration-dependent manner. Dehydroleucodine arrested the cell cycle at the G0/G1 phase, increased p27 and decreased both cyclins A and D and their partners (e.g., CDK2 and CDK4). Furthermore, DhL downregulated expression of histone demethylase JMJD2 as well as repressed the expression of histone methyltransferase MLL4, which in turn diminished the expression of C/EBPb and PPARg, respectively. 11,13-dihydro-dehydroleucodine blocked the accumulation of lipid droplets and inhibited the expression of PPARγ and C/EBPβ. Collectively, the results indicate that the inhibition of early stage preadipocyte differentiation by DH-DhL may be associated with cell cycle arrest at the G0/G1 phase. Dehydroparashin-B significantly decreased the accumulation of lipid content and downregulated the expression of CEBPβ, PPARγ and CEBPα as well as FAS. Interestingly, the addition of DhP inhibited the number as well as the size of the lipid droplets during the differentiation of 3T3-L1 preadipocytes. Taken together, this data suggests that DhP has an important inhibitory effect on cellular pathways regulating adipocyte differentiation.

Made available in DSpace on 2015-05-14T12:59:50Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 9147230 bytes, checksum: 092aa2337723ed50987e1d968d4fd438 (MD5) Previous issue date: 2013-04-26 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The family Celastraceae is distributed in tropical and subtropical regions, including North Africa, South America and Asia, particularly in China. It consists of 98 genera and about 1200 species, and in Brazil, this family is represented by four genera: Maytenus Juss., Austroplenckia Lund., Franhofera Mart. and Salacia Mart. The genus Maytenus is one of the largest family Celastraceae, and in Brazil, 76 species are recognized. Maytenus distichophylla Mart. Reissek former is used in folk medicine to treat stomach ulcers and is well adapted to semi-arid Northeast of Brazil. There are no reports of phytochemical and pharmacological study with this species. The aim of this work was to contribute to the phytochemical study of plants of the Brazilian Northeast through the isolation and identification of chemical constituents from leaves and roots of Maytenus distichophylla. Was used for both adsorption and partition chromatography on columns, thin layer chromatography on analytical and preparative. M. distichophylla was collected in the municipality of Maturéia-PB, and identified by Prof.. Dr. Maria de Fátima Agra. The powder obtained from the leaves and roots was subjected to maceration with methanol for 72 hours to yield the methanolic solution which was concentrated on rotaevaporator to give the methanol extract. Leaf extract was partitioned generating phases hexane, chloroform and ethyl acetate. The chloroform phase and ethyl acetate was subjected to chromatographic procedures known pentacyclic triterpenes six isolating the series friedelano, 3β-friedelinol, fridelina 3-oxo-12α-hidroxifriedelano 3-oxo-29α-hidroxifriedelano 3-oxo-30β-hidroxifriedelano and a new natural product the 6β, 12α-dihydroxy-friedelan-1 en-3,16,21-trione called maytensifolona. The chloroform phase roots of M. distichophylla was subjected to Medium Pressure Liquid Chromatography. The group of fractions 115-121 was subjected to High Performance Liquid Chromatography obtaining two sesquiterpene pyridine alkaloids known to wilforina and ebinifolina. Thus, these data corroborate the chemical composition of other species of Maytenus and contribute to the chemical knowledge of M. distichophylla. / A família Celastraceae distribui-se nas regiões tropical e subtropical, incluindo no norte da África, América do Sul e Ásia, particularmente na China. É constituída por 98 gêneros e aproximadamente 1200 espécies, sendo que no Brasil, esta família é representada por quatro gêneros: Maytenus Juss., Austroplenckia Lund., Franhofera Mart. e Salacia Mart.. O gênero Maytenus é um dos maiores da família Celastraceae e, no Brasil, são reconhecidas 76 espécies. Maytenus distichophylla Mart. ex Reissek é utilizada na medicina popular para o tratamento de úlceras estomacais e é bem adaptada ao semi-árido nordestino do Brasil. Não há nenhum relato de estudo fitoquímico e farmacológico com essa espécie. O objetivo desse trabalho foi contribuir com o estudo fitoquímico de plantas do Nordeste Brasileiro através do isolamento e identificação dos constituintes químicos das folhas e raízes de Maytenus distichophylla. Para tanto foi utilizado cromatografias de adsorção e partição em colunas, cromatografias em camada delgada analítica e preparativa. M. distichophylla foi coletada no município de Maturéia- PB, e identificada pela Prof. Dra. Maria de Fátima Agra. O pó obtido das folhas e raízes foi submetido à maceração com metanol durante 72 horas, obtendo-se a solução metanolica, que foi concentrada em rotaevaporador obtendo-se o extrato metanolico. O extrato das folhas foi particionado gerando as fases hexanica, clorofórmica e acetato de etila. A fase clorofórmica e acetato de etila foi submetida a procedimentos cromatograficos isolando seis triterpenos pentacíclicos conhecidos da serie friedelano, 3β-friedelinol, fridelina, 3-oxo-12α-hidroxifriedelano, 3-oxo-29-hidroxifriedelano, 3-oxo-30β-hidroxifriedelano e um novo produto natural o 6β,12α-dihidroxi-friedelan-1en-3,16,21-triona denominado maytensifolona. A fase clorofórmica das raízes de M. distichophylla, foi submetida a Cromatografia Liquida de Média Pressão. O grupo de frações 115-121 foi submetida a Cromatografia Liquida de Alta Eficiência obtendo-se dois alcaloides sesquiterpenos piridinicos conhecidos a wilforina e ebinifolina. Dessa forma, esses dados corroboram com a constituição química de outras espécies de Maytenus e contribuem para o conhecimento químico de M. distichophylla.

A química de produtos naturais sempre foi uma fonte importante de novas substâncias e de substâncias bioativas. No mundo moderno, o homem utiliza os produtos naturais para diversos fins: corantes, edulcorantes, essências, defensivos agrícolas e principalmente medicamentos. Com o desenvolvimento das técnicas de isolamento de substâncias, cresceu a necessidade de organizar as informações obtidas e também a criação de meios para a identificação mais rápida das substâncias isoladas. Esta foi uma das necessidades que fez surgir a Quimioinformática. Quimioinformática é uma disciplina que utiliza os métodos da informática para organizar dados químicos, analisar estes dados e gerar novas informações a partir destes dados. Esta ferramenta tem sido utilizada com sucesso em procura por novas drogas (QSAR/QSPR), elucidação estrutural automatizada de substâncias orgânicas e em cálculos e previsão de propriedades físico-químicas de diversas moléculas. Os objetivos do presente trabalho foram o estudo fitoquímico de espécies dos gêneros Dimerostemma e Ichthyothere com o intuito de isolar novas substâncias e o desenvolvimento de técnicas envolvendo quimioinformática com o intuito de auxiliar a elucidação estrutural de produtos naturais. Realizou-se a técnica de microamstragem de tricomas glandulares de diversas espécies pertencentes a gêneros da tribo Heliantheae (Viguiera, Tithonia, Dimerostemma). Através da microamostragem foi possível identificar diversas substâncias presentes nos tricomas glandulares das espécies analisadas. Das duas espécies de Dimerostemma investigadas (D. brasilianum e D. rotundifolium) foi possível identificar dois germacrolidos e dois eudesmanolidos, enquanto que de Ichthyothere terminalis foi possível a identificação de dois melampolidos, todos eles lactonas sesquiterpênicas. Foram treinadas redes neurais artificiais para a realização da identificação dos esqueletos carbônicos de determinadas substâncias a partir dos dados obtidos através dos espectros de RMN 13C, sendo que os resultados obtidos podem ser considerados satisfatórios. Foi desenvolvido um software para efetuar a identificação automática de substâncias através da comparação com uma biblioteca de padrões que possui dados cromatográficos de 51 lactonas sesquiterpênicas. Esse software, chamado de NAPROSYS, também é capaz de fazer comparação de dados de RMN de amostra com dados de RMN presentes em uma biblioteca de dados, tornando possível a identificação imediata de substâncias presentes na biblioteca e também auxiliar a elucidação estrutural de substâncias que não estão nela presentes. Para testar a eficiência do NAPROSYS, o programa foi utilizado com sucesso para identificar LSTs através da microamostragem de tricomas glandulares. A eficiência do NAPROSYS em identificar dados de RMN de substâncias presentes na biblioteca foi testada com substâncias isoladas do gênero Tithonia e Viguiera que possuem substâncias bem descritas na literatura e já isoladas no nosso laboratório, sendo que os resultados apresentados foram excelentes. Criou-se também dois modelos de redes neurais para prever tempos de retenção de lactonas sesquiterpênicas em cromatografia líquida (QSRR) com o objetivo de melhorar o desempenho do NAPROSYS em análises de dados cromatográficos. Os resultados para este caso, embora coerentes, precisam ser melhorados. Neste trabalho concluimos que o uso das técnicas clássicas juntamente com as novas técnicas de Quimoinformática pode se tornar uma ferramenta muito eficaz para a elucidação estrutural e busca de substâncias com determinadas propriedades químicas ou mesmo na bioprospecção de novas substâncias bioativas. / Natural products chemistry has always been an important source for new andbioactive compounds. In modern world, mankind uses natural products to do many tasks: colouring, as essences, as agricultural defensives and many as medicines. Within the development of compound isolation techniques, the need for information organisation has grown. The need for quickly identification of isolated compounds has also grown. This was one of the necessities that made Chemoinformatics emerge. Chemoinformatics is a discipline that uses informatics as a tool to organise, analise and to generate new knowledge from chemical data. This tool has been used with success in automate structure elucidation, drug development (QSAR/QSPR) and to predict chemical-physical data of many molecules. The aims of the present work were the phytochemical study of species of the genera Dimerostemma and Ichthyothere to isolate new compounds, and the development of chemoinformatics techniques to aid natural products structure elucidation. The glandular trichome microsampling was made for diverse species of genera from the tribe Heliantheae (Viguiera, Tithonia, Dimerostemma). Many compounds were identified through glandular trichome microsampling. Two germacrolides and two eudesmanolides were identified from Dimerostemma species (D. brasilianum and D. episcopale), while from Ichthyothere terminalis two melampolides were identified, all of them being sesquiterpene lactones. Artificial Neural Networks were trained to make skeleton identification from data obtained from 13C NMR and the obtained results can be considered satisfactory. A software was developed to make automatic compound identification through the comparation with a compound library that possesses data from 51 STLs. This software is called NAPROSYS is also able to compare the NMR data of the sample with the NMR data stored into a compound library, making the imediate identification of compounds present into library possible and also help the structure elucidation of unknown compounds. To test NAPROSYS\' efficience to identify NMR data of compunds sored into the library was made with compounds isolated from species of Tithonia and Viguiera genera, because these genera has well describe compounds in the literature and that has been isolated in our laboratory, and the obtained results are excellent. Two Artificial Neural Network models were created to predict the retention time of sesquiterpene lactones in liquid cromatography (QSRR) with the aim of improve NAPROSYS performance in cromatographic data analysis. The results for this case, although coherent, can be improved. The conclusion of this work is that the use of classical techniques with the new techniques of chemoinformatics can be a very efficient tool to make structure elucidation, search for compounds with certain chemical properties and even the search for new bioactive compounds.

A presente dissertação descreve um estudo químico das cascas do tronco de Guarea guidonia (L.) Sleumer (Meliaceae). Do extrato hexânico de cascas coletadas de um espécimen localizado em Campo Grande (MS) foram isolados e identificados cinco sesquiterpenos: o trans 1(10)-epóxi-4(15)-cariofileno; o 1(10)-epóxi-4,7-humuladieno; o 10-aloaromadendranol (viridiflorol); o 1(10),4-diepóxi-7-humuleno e o 3-oxa-10-aloaromadendranol e quatro esteróides: o sitosterol, o estigmasterol, o palmitato do sitosterol e o oleato do sitosterol. Esteróides glicosilados foram também detectados nesse extrato. Uma destilação por arraste a vapor d\'água forneceu o óleo essencial que foi comparado por cromatografia gasosa, com o óleo extraído de cascas de uma planta localizada em Araraquara (SP). Os cromatogramas dos dois óleos indicam que eles possuem composições químicas muito semelhantes. Do óleo essencial obtido do espécimen de Araraquara foram identificados onze sesquiterpenos: o 1(10),4(15),5-germacratrieno (germacreno D), o cis 4,10(14)-cadinadieno (γ-muuroleno), o trans 4,10(14)-cadinadieno, o 1(10),4-cadinadieno (σ-cadineno), o 1(10),4(15)-cariofiladieno, o 1,3,11-elematrieno (β-elemeno), o 1(10),4,7-humulatrieno (α-humuleno), o cis e o trans 1(10)-epóxi-4(15)-cariofileno, o 1(10)-epóxi-4,7-humuladieno e o 10-aloaromadendranol (viridiflorol). Os três últimos sesquiterpenos foram encontrados também no extrato hexânico do espécimen que vive em Campo Grande. As substâncias foram isoladas através de cromatografias de adsorção em sílica gel e desta impregnada com AgNO3 e identificadas por espectroscopias de RMN de 1H e de 13C (PND e DEPT 1350) e de massas. / This work describes a chemical study of the bark wood of Guarea guidonia (L.) Sleumer (Meliaceae). The hexanic extract of barks collected from an especimen growing at Campo Grande (MS) afforded five sesquiterpenes: the trans 1(10)-epoxi-4(15)-cariofileno; the 1(1 0)-epoxi-4,7-humuladieno; the 10-aloaromadendranol (viridiflorol); the 1(10),4-diepoxi-7-humuleno and the 3-oxa-10-aloaromadendranol and four steroids: the sitosterol, the stigmasterol, the palmitato of sitosterol and the oleato of sitosterol. From this extract were also detected glycosyl steroids. The essencial oil obtained from a steam water extraction of this barks was compared with that extract fiom the barks of an Araraquara\'s plant (SP). The chromatograms of the oils showed that the chemical composition of both oils are very similar. From the Araraquara\'s oil were identified eleven sesquiterpenes: the 1(10),4(15),5-germacratrieno (germacreno D), the cis 4,10(14)-cadinadieno (γ-muuroleno), the trans 4,10(14)-cadinadieno, the 1(10),4-cadinadieno (σ-cadineno), the 1(10),4(15)-cariofiladieno, the 1,3,11-elematrieno (β-elemeno), the 1(10),4,7-humulatrieno (α-humuleno), the cis and the trans 1(10)-epoxi-4(15)-cariofileno, the 1(10)-epoxi-4,7-humuladieno and the 10-aloaromadendranol (viridiflorol). The last three sesquiterpenes were also isolated fiom the hexanic extract. The compounds were isolated by adsortion chromatography on silica gel and on silica gel impregnated by AgNO3 and identified by 1H and 13C NMR (PND and DEPT 1350) and mass spectroscopy.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2017-07-04T14:51:48Z No. of bitstreams: 1 jordanadamascenogitiranadesantana.pdf: 3160925 bytes, checksum: 910c9f83a7dacd60918a4704f0d0fdeb (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-08-08T13:39:58Z (GMT) No. of bitstreams: 1 jordanadamascenogitiranadesantana.pdf: 3160925 bytes, checksum: 910c9f83a7dacd60918a4704f0d0fdeb (MD5) / Made available in DSpace on 2017-08-08T13:39:58Z (GMT). No. of bitstreams: 1 jordanadamascenogitiranadesantana.pdf: 3160925 bytes, checksum: 910c9f83a7dacd60918a4704f0d0fdeb (MD5) Previous issue date: 2015-07-30 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais / Vernonia polyanthes Less. (Asteraceae), popularmente conhecida como assa-peixe, é uma espécie vegetal nativa da América do Sul, principalmente do Brasil. Esta planta medicinal é tradicionalmente utilizada em casos de gripes, resfriados, tosses, febre, bronquite, contusões, hemorroidas, infecções do útero e no tratamento de malária. Farmacologicamente, V. polyanthes tem sido relatada como detentora de propriedades anti-hipertensiva, antinociceptiva, anti-ulcerogênica, anti-inflamatória, leishmanicida e antimicrobiana. Estudos fitoquímicos focando esta espécie, disponibilizados na literatura, relatam a presença de metabólitos secundários pertencentes, principalmente, às classes dos flavonoides e terpenos, especialmente lactonas sesquiterpênicas. O presente estudo descreve a avaliação da atividade antibacteriana do extrato de lavagem foliar (extrato VP) e da lactona sesquiterpênica glaucolídeo A isolada de V. polyanthes, no intuito de corroborar cientificamente com o uso tradicional dessa espécie vegetal e de verificar a potencialidade do extrato VP e desta molécula como prováveis fitoterápico e protótipo de um agente antibacteriano, respectivamente. O extrato VP foi obtido por meio da lavagem das folhas íntegras e secas de V. polyanthes com diclorometano e o glaucolídeo A isolado a partir do fracionamento desse extrato por métodos de cromatografia em coluna. A atividade antibacteriana foi investigada por meio da determinação da Concentração Inibitória Mínima (CIM) pelo método de microdiluição em caldo, segundo recomendações do Clinical Laboratory Standards Institute (CLSI). Em seguida, procedeu-se ao estabelecimento da Concentração Bactericida Mínima (CBM) e à classificação do efeito farmacológico bactericida ou bacteriostático. Staphylococcus aureus ATCC® 6538™/ATCC® 29213™, Escherichia coli ATCC® 10536™/ ATCC® 25922™, Salmonella enterica subsp. enterica serovar Choleraesuis ATCC® 10708™, Salmonella enterica subsp. enterica serovar Thyphimurium ATCC®13311™ e Pseudomonas aeruginosa ATCC® 9027™/ATCC® 27853™ foram usadas como cepas de referência. O extrato VP revelou atividade antibacteriana frente a 5 das 8 cepas testadas, sendo mais ativo contra S. aureus ATCC® 6538™ e ATCC® 29213, com valores de CIM iguais a 0,625 mg/mL, refletindo efeito bacteriostático e CBM de 2,5 mg/mL e 1,25 mg/mL, respectivamente. O glaucolídeo A apresentou atividade antibacteriana frente às referidas cepas de S. aureus, com valores de CIM de 250 µg/mL e 500 µg/mL, efeito bacteriostático e CBM de 500 µg/mL e > 500 µg/mL, nessa ordem, sendo inativo contra as demais cepas nas concentrações testadas. Os resultados do presente estudo sugerem que V. polyanthes e glaucolídeo A apresentam potencial antibacteriano, especialmente frente à S. aureus, sendo a atividade do extrato VP resultante, provavelmente, de um sinergismo entre essa lactona sesquiterpênica e outros metabólitos secundários presentes nessa planta medicinal. / Vernonia polyanthes Less. (Asteraceae), popularly known as "assa-peixe", is a native species of South America, especially in Brazil. This medicinal plant is traditionally used in cases of cold, flu, cough, fever, bronchitis, bruise, hemorrhoid, uterus infection and malaria's treatment. Pharmacologically, V. polyanthes has been reported as anti-hypertensive, antinociceptive, anti-ulcerogenic, anti-inflammatory, leishmanicidal and antimicrobial. Phytochemical studies focusing this plant, available in the literature, report the presence of secondary metabolites mainly belonging to the flavonoids and terpene classes, especially sesquiterpene lactones. The present study investigated the antibacterial activity of the leaf rinse extract (VP extract) and the sesquiterpene lactone glaucolide A isolated from V. polyanthes in order to scientifically corroborate with the traditional use of this plant species and to verify the potential of VP extract and this molecule as probable herbal and an antibacterial agent prototype, respectively. VP extract was obtained by washing the dried intact leaves of V. polyanthes with dichloromethane and the glaucolide A, isolated from the VP extract fractionation by column chromatography methods. Antibacterial activity was established by the determination of the Minimal Inhibitory Concentration (MIC) using the microdilution method according to the Clinical Laboratory Standards Institute (CLSI) guidelines followed by the Minimal Bactericidal Concentration (MBC), classifying the pharmacological effect as bacteriostatic or bactericidal. Staphylococcus aureus ATCC® 6538™/ATCC® 29213™, Escherichia coli ATCC® 10536™/ATCC® 25922™, Salmonella enterica subsp. enterica serovar Choleraesuis ATCC® 10708™, Salmonella enterica subsp. enterica serovar Thyphimurium ATCC® 13311™ and Pseudomonas aeruginosa ATCC® 9027™/ATCC® 27853™ were used as reference bacterial strains. The VP extract revealed antibacterial activity against 5 of 8 tested strains, being more active against S. aureus ATCC® 6538™ and ATCC® 29213™ with MIC values of 0.625 mg/mL, demonstrating bacteriostatic effect and MBC of 2.5 mg/mL and 1.25 mg/mL, respectively. Glaucolide A showed antibacterial activity against S. aureus with MIC values of 250 µg/mL and 500 µg/mL and, revealing bacteriostatic effect and CBM of 500 µg/mL and > 500 µg/mL, in this order, being inactive against the other strains at the concentrations tested. Our results suggest the antimicrobial potential of V. polyanthes and glaucolide A, notably against S. aureus, and the VP extract activity is probably due to the synergism between this sesquiterpene lactone and other secondary metabolites of this medicinal plant.

This thesis investigated the phytochemistry and pharmacology of immunomodulatory medicinal plant species used traditionally by the Q’eqchi’ Maya Healers Association (QMHA) of Belize, and the Cree of Eeyou Istchee (CEI) of northern Quebec. Using quantitative ethnobotanical methodology, we identified 107 plant species belonging to 49 families used by Q’eqchi’ healers in the treatment of symptoms from 14 usage categories related to inflammation. Regression analysis revealed that the Piperaceae, Araceae, and Begoniaceae are preferentially selected by the Maya. Healer consensus for plant species was high, with 56 species (52%) being used by all the healers, and consensus for usage categories was also high, as informant consensus factor (FIC) values for each category were greater than 0.4. Fifty-two Belizean species were evaluated for their TNF-α inhibitory activity in an LPS-stimulated THP-1 monocyte model. Twenty-one species (40%) demonstrated significant TNF-α inhibition when assayed at 100 µg/mL, 8 of which had greater than 50% of the activity of the parthenolide positive control (10 µg/mL). Significant regressions were found between the anti-inflammatory activity and total healer frequency of use (Fuse) and the use reports for 3 usage categories, which indicated that ethnobotanical parameters can in part predict the activity of traditionally used species. Five sesquiterpene lactones were isolated from the leaves of Neurolaena lobata, one of the most active species tested, all of which demonstrated anti-inflammatory activity greater than that of parthenolide (IC50 = 4.79 µM), with IC50s ranging from 0.17-2.32 µM. Lobatin B was the most active isolate tested. Ethanolic and water extracts of 17 species used by Cree healers were evaluated for their immunomodulatory activity. In general, the average anti-inflammatory activity of ethanolic extracts was 1.8 times greater than that of water extracts, and the pro-inflammatory activity of water extracts was 3.7 times greater than ethanolic extracts. Picea mariana and Pinus banksiana were the most anti-inflammatory ethanolic and water extracts, while the water extract of Sarracenia purpurea was the most pro-inflammatory. Picea marina cones, the most anti-inflammatory Cree medicine, were subjected to bioassay guided isolation. This led to the isolation of the anti-inflammatory lignan (+)-lariciresinol-9’-p-coumarate, which had an IC50 of 28.4 µM. Together, these results validate the traditional knowledge shared by our Q’eqchi’ and Cree collaborators, and draw attention to the therapeutic potential of subtropical and boreal plants as culturally appropriate complements to modern medicine. - Cette thèse porte sur la phytochimie et la pharmacologie des espèces de plantes médicinales immunomodulatrices utilisées traditionnellement par le Q’eqchi’ Maya Healers Association (QMHA) du Belize, et les Cris d'Eeyou Istchee (CEI) du nord du Québec. En utilisant une méthodologie ethnobotanique quantitative, nous avons identifié 107 espèces végétales appartenant à 49 familles utilisées par les guérisseurs Q'eqchi’ dans le traitement de symptômes appartenant à 14 catégories d'utilisation liées à l'inflammation. Une analyse de régression a révélé que les familles Piperaceae, Araceae, et Begoniaceae sont préférentiellement choisis par les Mayas. Le consensus entre guérisseurs pour les espèces végétales était élevé, avec 56 espèces (52%) étant utilisés par tous les guérisseurs, et le consensus pour les catégories d'utilisation était également élevé, car les valeurs de facteur de consensus des informants (FIC) pour chaque catégorie étaient supérieurs à 0,4. Cinquante-deux espèces du Belize ont été évaluées pour leur activité inhibitrice de TNF-α dans un modèle de THP-1 monocytes stimulés par le LPS. Vingt-et-une espèces (40%) ont montré une inhibition significative de TNF-α lorsque dosés à 100 µg/mL, dont 8 d’entre elles ont démontrées plus de 50% de l'activité du contrôle positif parthénolide (10 µg/mL). Des régressions significatives ont été observées entre l'activité anti-inflammatoire et la fréquence d'utilisation de guérisseurs totale (Fuse) et les rapports d'utilisation pour 3 catégories d'utilisation, ce qui indique que les paramètres ethnobotaniques peuvent en partie prédire l'activité des espèces traditionnellement utilisées. Cinq lactones sesquiterpéniques ont été isolés à partir des feuilles de Neurolaena lobata, l'une des espèces les plus actives testées, qui a démontré une activité anti-inflammatoire supérieure à celle du parthénolide (CI50 = 4,79 µM), avec des CI50 allant de 0,17 à 2,32 µM. Lobatin B était l’isolât le plus actif testé. Des extraits éthanoliques et aqueux de 17 espèces utilisées par les guérisseurs Cris ont été évalués pour leur activité immunomodulatrice. En général, l'activité anti-inflammatoire moyenne des extraits éthanoliques était 1,8 fois supérieure à celle des extraits d'eau, et l'activité pro-inflammatoire des extraits d'eau était de 3,7 fois supérieure à celle des extraits éthanoliques. Picea mariana et Pinus banksiana étaient les extraits éthanoliques et aqueux avec le plus d’activité anti-inflammatoire, tandis que l'extrait aqueux de Sarracenia purpurea était le plus pro-inflammatoire. Le cône de Picea marina, le médicament traditionnelle Cris le plus anti-inflammatoire, a été soumis à une isolation guidée par essais biologiques. Cela a mené à l'isolement du lignane anti-inflammatoire (+)-lariciresinol-9'-p-coumarate, qui avait une CI50 de 28,4 µM. Ensemble, ces résultats valident les connaissances traditionnelles partagées par nos collaborateurs Q'eqchi' et Cris, et mettent en évidence le potentiel thérapeutique des plantes subtropicales et boréales comme des compléments à la médecine moderne qui sont culturellement appropriées.

Ce mémoire décrit les travaux qui ont été réalisés sur la synthèse de l’hodgsonox, un sesquiterpène tricyclique comportant un éther diallylique dans un cycle tétrahydropyranique. Les approches envisagées sont la formation du cycle à cinq puis la formation du tétrahydropyrane et une autre plus convergente qui implique la synthèse des deux cycles en une seule étape. La première partie du mémoire discute de la synthèse d’un précurseur acyclique du cycle à cinq membres, afin de réaliser une réaction de métathèse de fermeture de cycle. Toutefois, les essais n’ont pas été concluants et cette voie a été abandonnée. Dans la deuxième partie, une nouvelle approche impliquant la synthèse d’un bicycle par une réaction de Pauson-Khand a été étudiée. Le précurseur de la réaction de Pauson- Khand a été préparé en 9 étapes (30% de rendement global) à partir du diéthyle tartrate. Le produit de cyclisation a été également obtenu mais il n’a pas été possible par la suite d’introduire le groupement isopropyle. Dans la dernière partie de ce mémoire, les travaux de Lise Brethous sur la synthèse de l’hodgsonox ont été repris. Celle-ci avait montré que le cycle à 5 membres pouvait être obtenu à partir de l’a-acétyl g-butyrolactone et que la formation du bicycle pouvait être réalisée par une réaction catalytique d’insertion d’un composé diazoïque dans un lien O-H. Certaines de ces étapes ont été optimisées et différents tests ont été effectués pour réaliser les dernières étapes de la synthèse de l’hodgosonox, mais sans succès. / This master thesis describes various strategies towards the total synthesis of (+)- and (-)-hodgsonox, a tricyclic sesquiterpene containing a diallylic ether functionality fused into a tetrahydropyran ring. The first synthetic approach involved the formation of the fivemembered ring followed by the formation of the tetrahydropyran moiety. The second approach consisted in the synthesis of both carbon cycles in a single stage. In the first part, is described the synthesis of an acyclic precursor to assemble the fivemembered ring via a ring-closing metathesis reaction. This approach proved unsuccessful and was not pursued. In the second part, is reported a Pauson-Khand cyclization to produce the bicyclic compound. The Pauson-Khand precursor was synthesized in 9 steps (30% overall yield) from diethyl tartarate. The cyclization was successful, but the isopropyl moiety could not be introduced. A former Lebel’s PhD student, Lise Brethous had also worked on the total synthesis of hodgsonox. She established that the 5-membered ring could be prepared from a-acetyl gbutyrolactone and observed the formation of the tetrahydropyran moiety via an OH-insertion reaction with a diazo intermediate. In the last part of this master thesis, some steps from Brethous’s approach were optimized. Unsuccessful attempts to tackle the last steps of the synthesis are also described.