Submitted by Natalie Mendes (nataliermendes@gmail.com) on 2015-07-28T00:17:52Z No. of bitstreams: 1 Disserta??o - C?ssio Santana.pdf: 10345998 bytes, checksum: 41cc3a308f7a72f1c4e10cc1ee2fe345 (MD5) / Made available in DSpace on 2015-07-28T00:17:52Z (GMT). No. of bitstreams: 1 Disserta??o - C?ssio Santana.pdf: 10345998 bytes, checksum: 41cc3a308f7a72f1c4e10cc1ee2fe345 (MD5) Previous issue date: 2014-02-05 / Funda??o de Amparo ? Pesquisa do Estado da Bahia - FAPEB / Chagas disease is a zoonosis caused by the hemoflagellate protozoan Trypanosoma cruzi, which affects millions of people in Latin America. Disease treatment is based on the use of two drugs, benznidazole and nifurtimox, which present low cure rates in the chronic phase of the disease as well as generating a number of adverse side effects. In this context, the development of new therapies for a better treatment of Chagas disease is necessary. In this study we investigated the anti-T.cruzi potential of physalins and betulinic acid, plant-based isolated steroids, in addition to synthetic derivatives from the latter, in in vitro assays. Our results demonstrate a high trypanocidal activity of physalins B and F and of the derivative BA5 against trypomastigote forms and on the processes of invasion and development of these on peritoneal macrophages. Electron microscopy revealed that physalin B or BA5 derivative treatment resulted in ultrastructural changes in the plasma membrane, Golgi apparatus, kinetoplast and endoplasmic reticulum of trypomastigotes forms. Additionally, these compounds contributed to the formation of atypical vacuoles in the appearance of myelin figures, which were labeled with MDC to confirm their identity as autophagic vacuoles. Flow cytometry analysis revealed that parasite death occurred mainly by necrosis. When combined with benznidazole, physalin B, physalin F or BA5 derivative, resulted in a higher anti-T. cruzi activity against amastigotes when compared to the compounds tested alone. These results indicate that steroids, such as physalins B and F and the derivative BA5, are potential candidates for an alternative treatment of Chagas disease. / A doen?a de Chagas ? uma zoonose causada pelo protozo?rio hemoflagelado Trypanosoma cruzi, que afeta milh?es de pessoas na Am?rica Latina. O tratamento dessa enfermidade se baseia na utiliza??o de dois f?rmacos, o benzonidazol e o nifurtimox, que possuem baixa taxa de cura na fase cr?nica da doen?a, al?m de gerarem uma s?rie de efeitos colaterais. Nesse contexto, o surgimento de novos medicamentos para uma quimioterapia mais adequada da doen?a de Chagas torna-se necess?rio. Neste estudo investigamos o potencial anti-T. cruzi de esteroides isolados de plantas, fisalinas e ?cido betul?nico, assim como de derivados sint?ticos deste ?ltimo, em ensaios in vitro. Nossos resultados demonstram uma elevada atividade tripanocida das fisalinas B e F e do derivado BA5 contra formas tripomastigotas e sobre os processos de invas?o e desenvolvimento destas em macr?fagos peritoneais. Atrav?s de ensaios de microscopia eletr?nica foi poss?vel observar que o tratamento com a fisalina B ou com o derivado BA5 causa altera??es ultraestruturais na membrana plasm?tica, complexo de Golgi, cinetoplasto e ret?culo endoplasm?tico das formas tripomastigotas, al?m da forma??o de vac?olos at?picos e o aparecimento de figuras miel?nicas, que foram marcadas com MDC para confirmar a sua identidade como vac?olos autof?gicos. An?lises por citometria de fluxo revelaram que a morte parasit?ria ocorre principalmente por necrose. A combina??o da fisalina B, da fisalina F ou do derivado BA5 com o benzonidazol resultou em uma maior atividade anti-T. cruzi frente a formas amastigotas quando comparados aos compostos testados de forma isolada. Estes resultados indicam que esteroides, tais como as fisalinas B e F e o derivado BA5, s?o potencias candidatos para o tratamento alternativo da doen?a de Chagas.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-04-03T19:41:30Z No. of bitstreams: 1 SarahPollyanaDiasDosSantos_DISSERT.pdf: 10111393 bytes, checksum: 44cc6b08e8b21238733661e330e1b811 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-04-05T19:31:53Z (GMT) No. of bitstreams: 1 SarahPollyanaDiasDosSantos_DISSERT.pdf: 10111393 bytes, checksum: 44cc6b08e8b21238733661e330e1b811 (MD5) / Made available in DSpace on 2017-04-05T19:31:53Z (GMT). No. of bitstreams: 1 SarahPollyanaDiasDosSantos_DISSERT.pdf: 10111393 bytes, checksum: 44cc6b08e8b21238733661e330e1b811 (MD5) Previous issue date: 2016-07-14 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Aspidosperma pyrifolium Mart., esp?cie nativa da caatinga com grande ocorr?ncia no nordeste, conhecida popularmente como pereiro, utilizada na medicina popular no tratamento de doen?as e enfermidades. Consta na literatura diversos trabalhos realizados, relatando a presen?a de classes de compostos org?nicos como alcaloides, entre outras, que seriam respons?veis por propriedades terap?uticas existentes nessa esp?cie. O estudo fitoqu?mico de A. pyrifolium, das fra??es da semente e cascas do fruto, utilizando m?todos cromatogr?ficos cl?ssicos e purifica??o por CLAE, nos permitiu o isolamento de tr?s compostos. A partir da fra??o hex?nica das cascas do fruto (APCF-H) foi isolado o triterpeno ?cido betul?nico, n?o relatado anteriormente na esp?cie A. pyrifolium. Das fra??es diclorometano (APSE/DCM) e acetato de etila 50% dicloromentano (APSEA/50% DCM) das sementes foi poss?vel o isolamento do alcaloide plumerano pentac?clico aspidospermina, do alcaloide ?-carbolina glicosilado Cordifolina, n?o descrito anteriormente na esp?cie e outro composto do tipo ?-carbolina que n?o possui uma estrutura elucidada at? o momento. O extrato das sementes fra??o diclorometano (APSE/DCM) foi analisado por CG/EM (cromatografia gasosa acoplada a espectrometria de massa) e seus dados foram comparados com dados na literatura em banco de dados (NIST08.LIB), foi identificada a presen?a dos alcaloides 1-acetil-aspidospermidina, aspidospermina e pirifolina, dentre outros compostos minorit?rios. A avalia??o da atividade antioxidante, pelo m?todo sequestrador de radical livre de DPPH (2,2-difenil-1picril-hidrazila), da fra??o APSE/DCM apresentou um percentual de atividade antioxidande m?ximo (AA% m?ximo) de 83,68%, com concentra??o inibit?ria 50% (IC50) 133,4867 ? 2,22. A fra??o das sementes fra??o diclorometano (APSE/DCM) foi avaliada em rela??o ? toxicidade contra Artemia salina, tendo em vista que j? existiam relatos da toxicidade nas folhas de A. pyrifolium, comprovamos a toxicidade aguda nas fra??o das sementes diclorometano com concentra??o letal m?dia (CL50) em 24 horas de 1,1524 ?g.ml-1, sendo consideradas t?xicas as esp?cies que apresentam CL50 inferior a 1000 ?g.ml-1. Na segunda parte do trabalho foi realizado um levantamento bibliogr?fico de alcaloides do tipo ?-carbolina de ocorr?ncia natural, com suas estruturas, atividades relatadas e um banco de dados com seus deslocamentos qu?micos em RMN 1H e 13C, totalizando 169 alcaloides descritos na literatura, do per?odo de 1992 a 2016, agrupados em quatro principais grupos, denominados ?-carbolina, dihidro-?-carbolina, tetrahidro-?-carbolina e os ?-carbolinas divergentes, este levantamento auxiliar? na elucida??o estrutural de novas mol?culas. / Aspidosperma pyrifolium Mart., a native species of the caatinga with great occurrence in the northeast, known perpularmente as pereiro, used in popular medicine in the treatment of diseases and diseases. There are several studies carried out in the literature, reporting the presence of classes of organic compounds such as alkaloids, among others, that would be responsible for the therapeutic properties of this species. The phytochemical study of A. pyrifolium, seed fractions and fruit peels using classical chromatographic methods and purification by HPLC allowed us to isolate three compounds. From the hexanic fraction of fruit peels (APCF-H) was isolated the betulinic acid triterpene, not previously reported in the A. pyrifolium species. From the dichloromethane (APSE/DCM) and ethyl acetate 50% dichloromethane (APSEA / 50% DCM) fractions of the seeds, it was possible to isolate the pentacyclic pentameric aspidospermine alkaloid from the glycosylated ?-carboline alkaloid Cordifolina, not previously described in the species and another compound of the ?-carboline type that does not have a structure elucidated until the moment. The extract of the seeds dichloromethane fraction (APSE/DCM) was analyzed by GC/MS (gas chromatography coupled to mass spectrometry) and its data were compared with data in the literature in database (NIST08.LIB), the presence of the alkaloids 1-acetyl-aspidospermidine, aspidospermine and pyrifoline, among other minor compounds. The antioxidant activity evaluation of the APSE/DCM fraction showed a percentage of maximum antioxidant activity (AA% maximum) of 83.68%, with the free radical sequestration method of DPPH (2,2-diphenyl-1-picryl-hydrazila). Inhibitory concentration 50% (IC50) 133.4867 ? 2.22. The fraction of the seeds dichloromethane fraction (APSE/DCM) was evaluated in relation to the toxicity against Artemia salina, considering that there were already reports of the toxicity in the leaves of A. pyrifolium, we verified the acute toxicity in the seed dichloromethane fraction with average lethal concentration (CL50) in 24 hours of 1.1524 ?g.ml-1, species with CL50 less than 1000 ?g.ml-1 being considered toxic. In the second part of the work a bibliographic survey of naturally occurring ?-carboline alkaloids with their structures, reported activities and a database with their chemical displacements in 1H and 13C NMR were carried out, totaling 169 alkaloids described in the literature. From 1992 to 2016, grouped into four main groups, named ?-carboline, dihydro-?-carboline, tetrahydro-?-carboline and the divergent ?-carbolines, this survey will aid in the structural elucidation of new molecules.