Made available in DSpace on 2016-04-28T20:15:26Z (GMT). No. of bitstreams: 1 2007-Renata da Silva Ribeiro.pdf: 705558 bytes, checksum: e16373fa17f99c734d58ede2a54543c6 (MD5) Previous issue date: 2007-02-27 / Ouratea cuspidata (Ochnaceae) crude extracts were added to experimental systems containing either host macrophages, or Leishmania braziliensis promastigotes (L566). The biological effect by the use of the hexane (OCH), methanol (OCMeH) and ethyl-acetate (OCAcEt) extracts added at different concentrations: 2mg/mL; 4mg/mL; 8mg/mL, 16mg/mL e 32mg/mL. The OCMeOH was the only fraction to which the anti parasitic potentiality was achieved at doses rates that generated bio protective effects in the host cells. The parasiticide action, due by direct exposure of promastigotes to OCMeOH, as well as the parasitic growth, and morphological alterations intermediary metabolism alterations (mitochondrial activity) were concomitantly measured. Biological macrophagic functions were also evaluated using Cricetus cricetus peritoneal macrophages as a model (Mf). Bio-protective assays were carried out in order to determine the free radical generation by the extract constituents. The results were compared with non treated L566 and Mf. 78% of the 16 mg/106 Mf/mL treated Mf were viable but a significant decrease of their phagocytosis capability (92%) was detected. Such alterations were evident in 62% of the cells treated with 32mg/106 Mf/mL. At this concentration cells were 70% viable and presented 98% phagocytosis suppression. The macrophages enzymatic- mitochondrial activity was gradually diminished, with 87% activity at exposures to 8mg/106 Mf/mL; 66,3% at exposures to 16mg/106 Mf/mL, and 49% at exposures to 32mg/106 Mf/mL, respectively. The anti parasitic effects were not associated to the promastigotes lack of viability. Instead, a significant rising on the L566 cells counting was detected when compared to control non treated parasites, from the first 6 hours treatment until 24 hours exposure. Morphological changes were detected in 62% of the 8mg/106 L566/mL parasite treated cells; 78% after 16mg/106 L566/mL treatment, and 98% of the cells presented morphological changes after 32mg/106 L566/mL treatment. Cells were either round-shaped, showing incomplete mitosis, or presenting double flagella, suggesting that the present extract containing substances that might interfere in the parasitic topoisomerase function. The parasitary mitochondrial activity evidenced the occurrence of a metabolic acceleration due by the OCMeOH treatment. The mitochondrial enzymatic activity was of 220% at exposures to 8mg/106 L566/mL; 389% at exposures to 16mg/106 L566/mL and of 480% at exposures to 32mg/106 L566/mL, respectively, when compared to the non treated parasites (100%). The anti parasitic potential (Therapeutic Index) was considered to be positive (acceptable) (TI=4,0) when estimated in function of the morphological changes observed at the extract concentration of 32mg/106 Mf/mL (LD62%), and at 8mg/106 L566/mL (ED62%), respectively. Since bi-flavonoids are the main constituents present in the OCMeOH, results were suggestive the anti parasitic effect was due to this group of secondary metabolites. As such biologically active molecules are known COX2 selective inhibitors, its internal use should be avoided. Otherwise, bi- flavonoids are good candidate substances to be applied as topical medicine to treat the American Tegumentar Leishmaniasis. / Os efeitos dos extratos de Ouratea cuspidata (Ochnaceae) foram avaliadas a partir da exposi??o de c?lulas hospedeiras e de promastigotas de Leishmania (Viannia) braziliensis (L566) ?s fra??es oriundas da sua parti??o hex?nica (OCH), metan?lica (OCMeOH) e acetato de et?lica (OCAcEt). Dentre estas, a fra??o OCMeOH foi a ?nica que apresentou potencial antiparasit?rio em doses que geraram bioprote??o aos sistemas hospedeiros. Assim, procederam-se an?lises de a??o parasiticida direta do extrato OCMeOH sobre formas promastigotas, onde, tamb?m, foram avaliados o crescimento, altera??es morfol?gicas e fun??es do metabolismo mitocondriais. Do mesmo modo, foram realizadas observa??es semelhantes em macr?fagos peritoneais de hamsters Cricetus cricetus (Mf) afim de se determinar a toxicidade relativa para o hospedeiro. Finalmente, foram realizados ensaios complementares de bioprote??o e gera??o de radicais livres, com a finalidade de confirmar a poss?vel aplica??o terap?utica dos constituintes presentes no extrato. Em todas as etapas experimentais, foram utilizadas cinco concentra??es de OCMeOH (2mg/mL; 4mg/mL; 8mg/mL; 16mg/mL e 32mg/mL), sendo que os resultados obtidos foram comparativamente avaliados em rela??o a sistemas de controle (L566 e Mf n?o tratados). Os (Mf) tratados com 16 mg/106 Mf/mL do extrato sofreram uma diminui??o significativa (92%) de sua capacidade fagocit?ria, mas permaneceram vi?veis, em sua maioria (78%). As altera??es morfol?gicas mostraram-se mais evidentes em 62% das c?lulas tratadas com 32mg/106 Mf/mL, com 70% de viabilidade e 98% de inibi??o da fagocitose. A atividade enzim?tica-mitocondrial macrof?gica apresentou diminui??o gradativa, com a preserva??o de 87% das fun??es enzim?ticas nas c?lulas tratadas com 8mg/106 Mf/mL; 66,3% daquelas tratadas com 16mg/106 Mf/mL e 49% das tratadas com 32mg/106 Mf/mL. As L566 n?o apresentaram a perda da viabilidade ap?s exposi??o de 24horas ?s diferentes concentra??es de OCMeOH. Ao contr?rio, houve um aumento significativo no n?mero de promastigotas que, a partir de seis horas de cultivo, mostraram-se sempre superiores ? quantidade de parasitos n?o tratados. Foram detectadas altera??es morfol?gicas em 62% das L566 tratadas com 8mg/106 L566/mL; 78% daquelas tratadas com 16mg/106 L566/mL e 98% das tratadas com 32mg/106 L566/mL. As promastigotas tratadas apresentaram-se arredondadas, com mitose incompleta, ou apresentando dois flagelos, sugerindo que as subst?ncias presentes no extrato podem interferir nas topoisomerases parasit?rias. A atividade mitocond rial parasit?ria evidenciou a ocorr?ncia de acelera??o metab?lica induzida pelo tratamento com OCMeOH. A atividade enzim?ticamitocondrial foi de 220% no tratamento com 8mg/106 L566/mL; de 389% com 16mg/106 L566/mL e 480% com 32mg/106 L566/mL quando comparadas ao controle (100%). O potencial antiparasit?rio (Indice Terap?utico) foi considerado positivo (IT=4,0) em fun??o das altera??es morfol?gicas observadas nas concentra??es de 32mg/106 Mf/mL (LD62%), e 8mg/106 L566/mL (ED62%). Sendo biflavon?ides os constituintes presentes em maior propor??o, atribuiu-se a este grupo de subst?ncias a a??o antiparasit?ria e imunot?xica observadas para OCMeOH. Os bi-flavon?ides presentes no extrato metan?lico de Ouratea cuspidata est?o entre os inibidores seletivos de COX2, sugerindo o desenvolvimento de pesquisas que visem sua utiliza??o t?pica em animais naturalmente infectados.

Doctor of Philosophy / Department of Chemistry / Duy H. Hua / I. Ovalicin is a natural product isolated from the culture of fungus Pseudorotium ovalis Stolk, it selectively inhibit type 2 methionine amino-peptidase (MetAP 2), which related to many physiological activities such as angiogenesis. Total synthesis of [plus or minus] ovalicin, its C4(S*), C4(S*)C5(S*) stereo-isomers, and C5 regio-isomer were synthesized via an intramolecular Heck reaction of (Z)-3-(t-butyldimethyl silyloxy)-1-iodo-1,6-heptadiene utilizing a catalytic amount of palladium acetate. Subsequent epoxidation, dihydroxylation, methylation (or stereochemistry inversion before or after methylation) and oxidation led to a variety of ketones, key intermediates for synthesis of ovalicin and its analogues. Introduction of side-chain to ketones by lithium (Z)-6-methylhepta-2,5-dien-2-ide and following functional group transformation led to ovalicin and its analogues. Anti-trypanosomal activities of various ovalicin analogues and synthetic intermediates were evaluated. II. Bio-based polymers from vegetable oils are renewable and environment-friendly materials. Dihydroxylated, trihydroxylate, tetrahydroxylated and hexahydroxylated triglycerides, triamino and triisopropylamino glycerides were synthesized from model triglyceride glyceryl trioleoate. These monomers were cross-linked with 1, 4-phenylene diisocynate to make polyurethanes and polyureas. The physical properties of these polymers were examined by gel content and swelling value measurements, thermodynamic and viscoelastic properties were studied from TGA, DSC and DMA measurements. The structure-property relationship was discussed based on these measurements. III. Diacetylenic fatty acids were widely applied in material science to regulate alignment on surface and stabilize self-assembled nanomaterials. A novel synthetic method of diacetylenic fatty acids from vegetable oils was developed. Its self-assembling properties on alumina surface were measured and discussed.