Our previous study identified both high p27kip1 and low CKS1B protein levels were independent prognosis markers in hepatocelular carcinomas (HCC), however, CKS1B overexpression implicated clinical aggressiveness of HCC but not p27Kip1 protein turnover. In this study, we demonstrated a cytotoxic compound, goniothalamin (GTN), that stabilized p27kip1 protein and downregulated CKS1B mRNA as well as protein levels in HCC-derived cell lines. The IC50 of TP53-negative Hep-3B and TP53-positive SK-Hep1 after treatment with GTN for 48 h were detected as 25 and 10 £gM, respectively. The GTN induced DNA damages, formation of £^H2AFX foci, and upregulation of £^H2AFX protein levels in dose-dependent manners in both cell lines. In addition, GTN arrested Hep-3B and SK-Hep1 cells in G2/M and G1 phases, respectively. Protein levels of several cell cycle regulators, including p27Kip1 in Hep-3B and, p21Cip1 and CKS1B in SK-Hep1 cells have been noticeable upregulated after treatment with GTN. In Hep-3B cells, GTN induced the configuration of p27Kip1/CCND1, p27Kip1/CDK2; p27Kip1/CCNB1 and p27Kip1/CDK1, however, repressed the p27Kip1/STMN1 complexes. On the other hand, GTN evidently induced the configuration of p21Cip1/CDK2 and p21Cip1/CCNE1, but repressed the p21Cip1/SKP2 and p21Cip1/CKS1B complexes, in SK-Hep1 cells. The CKS1B protein abundance that was induced by GTN was stemmed from the CKS1B transactivity. However, GTN-induced p27Kip protein profusion in Hep-3B cell line was, instead, due to the stabilization of p27Kip1 protein, mainly in the cytosol. Both GTN and the proteasome inhibitor, MG132, induced p27Kip1 ubiquitination and p27Kip1 protein profusion with an additive effect, regardless of CKS1B protein level, suggesting that the p27Kip1 protein might be regulated by GTN through an alternative ubiquitin-complex in Hep-3B cells. In addition to the quantitative reverse transcription -polymerase chain reaction, GTN-induced p21Cip1 protein level in SK-Hep1 cells could be traced back to mRNA level, by evidence of quantitative immunoprecipitation/chromatin immunoprecipitation assay with primer sets specific to two regions of the p21Cip1 proximal promoter. Taken together, GTN is able to induce the cell cycle arrest via stabilization of p27Kip1 protein and downregulation of CSK1B mRNA and subsequent protein levels in TP53-negative Hep-3B and TP53-positive SK-Hep1 cells, respectively. The GTN-repressed CKS1B transcription and thereafter, cell cycle arrest, was irrelevant to the p27Kip1 protein stability in Hep-3B cells.

Orientador: Ronaldo Aloise Pilli / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Química / Made available in DSpace on 2018-08-23T10:20:29Z (GMT). No. of bitstreams: 1 Barcelos_RosimeireCoura_D.pdf: 15482677 bytes, checksum: f8abc41bb1731100795495890fe0c509 (MD5) Previous issue date: 2013 / Resumo: Esse trabalho descreve a preparação de 48 compostos inéditos inspirados nas estruturas da goniotalamina (23) e da piplartina (210), bem como a avaliação da citotoxicidade in vitro desses derivados frente a uma variedade de linhagens de células tumorais humanas. Foram sintetizados 13 análogos da goniotalamina (23), os quais exibiram interessante perfil citotóxico, especialmente os compostos 144, 146 e 148 apresentando grupos metoxila no anel aromático da goniotalamina (23). Ainda, o análogo 164 com o grupo hidroxila em meta reduziu a viabilidade celular do câncer de pâncreas em 50% em uma concentração duas vezes menor em relação a goniotalamina (23). Também foi feita a substituição do anel lactônico por uma lactama, sendo observada uma diminuição significativa da atividade citotóxica dos 9 aza-análogos sintetizados. Em seguida, foi realizado um breve estudo de relação estrutura-atividade da piplartina (210), no qual se evidenciou a importância da carbonila a,b-insaturada endocíclica para a citotoxicidade dessa classe de compostos. Descreveu-se a preparação e a avaliação citotóxica in vitro de 21 derivados N-acilados dos aza-análogos. A N-acilação do nitrogênio lactâmico resultou em compostos ainda mais potentes do que a goniotalamina (23) e a piplartina (210) contra diversas linhagens de câncer, tal como o derivado 294 contendo o grupo N-crotonoila. Além disso, essas moléculas apresentaram um perfil citotóxico promissor contra as células tumorais PANC-1, uma vez que todos os análogos N-acilados foram mais ativos do que a goniotalamina (23). A piplartina (210) e os derivados 269 e 309 foram os mais potentes, sendo que o híbrido goniotalamina-piplartina 269 apresentou o menor valor de IC50, validando a estratégia de hibridação molecular / Abstract: This work describes the preparation of 48 novel compounds inspired on the structures of goniothalamin (23) and piplartine (210) as well as the in vitro assessment of their cytotoxicity against a variety of human tumor cell lines. Thirteen analogs of goniothalamin (23) were synthesized which exhibited interesting cytotoxic profiles, especially compounds 144, 146 and 148 bearing aromatic methoxyl groups. Analog 164 presenting a meta-hydroxyl group reduced the cell viability of pancreatic cancer by 50% at a concentration twice as low as the goniothalamin (23). Subsequently, the lactonic ring was substituted by a lactam, however leading to a significant reduction of the cytotoxic activity of the corresponding 9 aza-analogs. A brief structure-activity relationships study was performed, in which the importance of a,b-unsaturated endocyclic carbonyl group for cytotoxic activity of this class of compounds was demonstrated. The preparation and in vitro cytotoxic evaluation of 21 N-acylated aza-analogs derivatives is herein described. The N-acylation of the lactam nitrogen resulted in compounds that were more potent than goniothalamin (23) and piplartine (210) against diverse strains of cancer, such as compound 294 containing the N-crotonoyl group. Moreover, these molecules showed a promising cytotoxic profile against tumor cells PANC-1, since all the N-acylated analogs were more active than goniothalamin (23). Piplartine (210) and derivatives 269 and 309 were the most potent, and the goniothalamin-piplartine hybrid 269 displayed the lowest IC50 value of all, thereby validating the strategy of molecular hybridization / Doutorado / Quimica Organica / Doutora em Ciências

The goniothalamin (GTN) and related styryl-lactones were found to be cytotoxic and apoptotic to a variety of tumor cell lines including breast cancer MCF-7, cervical cancer HeLa and leukemia HL60. In HL60 cells, GTN triggers mitochondria dysfunction, caspase activation and eventually, apoptosis. In this study, we demonstrated that GTN was able to induce apoptosis in hepatocellular carcinoma derived cells, SK-Hep1 and Hep-3B cells via upregulation of the phorbol-12-myristate-13-induced 1 (PMAIP1) protein , alternation of mitochondrial membrane potentials (P < 0.05) via TP53-dependent and -independent transactivations. Treatment with GTN induced DNA damage, formation of reactive oxygen species (ROS, P < 0.05) and activated cleaved CASP8, CASP9, CASP3 and poly (ADP-ribose) polymerase 1 proteins. However, cleaved BH3 interacting domain (BID) death agonist protein was not identified, suggesting that an intrinsic cellular stress was produced after GTN treatments in both cell lines. A pan caspase inhibitor, z-VAD-FMK, suppressed GTN-induced apoptotic cell percentages (P < 0.05), demonstrating that GTN-induced apoptosis was mediated by the activation of the caspase cascade protein. The GTN induced ROS formation prior to DNA damage in SK-Hep1, yet in reverse order in Hep-3B cells. Moreover, GTN induced DNA damages through activation of the gamma H2A histone family member X (£^H2AFX, Ser139)/phospho-CHK1 checkpoint homolog (pCHEK1, Ser345)/phospho-CHK2 checkpoint homolog (pCHEK2, Thr68)/phosphos-tumor protein p53 (pTP53, Ser15), and £^H2AFX/pCHEK2 (Thr68), in SK-Hep1 and Hep-3B cells,respectively. Among five integral outer mitochondrial membrane proteins that blocks or induces apoptotic death, PMAIP1 protein and PMAIP1 mRNA levels were upregulated after GTN treatments for 4 to 6 h in both cell lines (P < 0.05). Transfection of shRNA interference plasmids targeting PMAIP1 gene downregulated PMAIP1 mRNA (P < 0.05) and PMAIP1 protein (P < 0.05) levels, as well as GTN-induced apoptotic cell percentages (P < 0.05) in both cell lines. In parallel, transfection of the shRNA interference plasmid targeting TP53 gene in SK-Hep1 cells, downregulated TP53 and PMAIP1 mRNA (P < 0.05) and TP53, pTP53, PMAIP1, cleaved PARP1 protein levels and apoptotic cell percentages (P < 0.05). Treatment with the TP53 inhibitor, PFT-£\ or transfection of a dominant negative TP53 plasmid, pTP53mt135, repressed TP53, pTP53 and PMAIP1 protein and/or TP53, PMAIP1 mRNA levels (P < 0.05), however, significantly augmented GTN-induced apoptotic cell percentages (P < 0.05). Cytosol/mitochondria fractionation identified that TP53, along with PMAIP1 proteins, were translocated to mitochondria after GTN treatment in time-dependent manners. Taken together, our studies demonstrated that GTN induced apoptosis via PMAIP1 via both TP53-dependent and -independent transactivation pathways. In TP53-positive SK-Hep1 cells, PMAIP1 and TP53 proteins conducted synergic effects.

The objective was to study apoptotic effects and underlying molecular mechanisms of goniothalamin (GTN) in hepatocellular carcinoma (HCC)-derived cells. The GTN that isolated from Goniothalamus amuyon, was found to possess profound cytotoxic activities against human SK-Hep1 and Hep-3B cells in vitro. The cytotoxicity of GTN/cell viability was measured by MTT assay at 570-nm absorbance and the IC50 at 72 h were determined as 7.5, 17 mM in SK-Hep1 and Hep-3B respectively. The GTN induced cell death and accumulation of reactive oxygen species in HCC-derived cells. One reactive oxygen species inhibitor, N-actylcysteine, further restored cell viabilities post-GTN treatments. Formation of £^-H2AX foci suggested that GTN-induced DNA damages were double-strand breaks. The GTN arrested cell cycle at G0/G1 by regulation of CCND, CCNE1, RB1 and E2F1 proteins in SK-Hep1, and at G2/M by regulation of CCNB1 in Hep-3B cells. The GTN-induced apoptosis in HCC-derived cells were evidenced by phosphatidylserine externalization and involved both extrinsic and intrinsic pathways in SK-Hep1 cells, but only extrinsic pathway in Hep-3B cells. In SK-Hep1 cells, GTN induced apoptosis and cell cycle arrest through TP53-mediated pathway in contrast to that of TP53/FAS mutated Hep-3B cells. Importantly, GTN was able to induce apoptosis in both TP53 wild type and TP53/FAS mutated HCC-derived cells. On the other hand, GTN was able to induce TP53 and p21WAF1/Cip1 up-regulation in SK-Hep1 and Hep-3B cells and p27Kip1 up-regulation in Hep-3B cells. These results demonstrated that GTN induced apoptosis in HCC cells through distinct signaling pathways.

Orientadores: João Ernesto de Carvalho, Ronaldo Aloise Pilli / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-09-28T18:41:42Z (GMT). No. of bitstreams: 1 Costa_DeboraBarbosaVendramini_D.pdf: 5920460 bytes, checksum: d6638ad4d3d0a96e4bbcdb55823f3254 (MD5) Previous issue date: 2012 / Resumo: A carcinogênese é um processo longo, multi-etapas, onde células normais progressivamente adquirem um fenótipo neoplásico. Sua origem é favorecida por fatores genéticos, exposição à carcinógenos químicos, infecções crônicas e incorporação de mutações em genes envolvidos com a regulação da homeostase celular. O crescente entendimento da biologia tumoral tem fornecido alvos moleculares para a triagem orientada de quimioterápicos e de agentes quimiopreventivos, geralmente de origem natural ou sintetizados com base em produtos naturais. A evolução da química orgânica permitiu que compostos naturais fossem sintetizados de forma econômica e aperfeiçoados quanto às suas propriedades físico-químicas, favorecendo sua disponibilidade. Esse trabalho teve como foco a goniotalamina, uma estiril-lactona naturalmente encontrada na forma enantiomérica (R) em plantas do gênero Goniothalamus (Annonaceae). Foram sintetizadas as formas (R), (S) e racêmica, que em painel de nove linhagens tumorais humanas levaram à inibição de crescimento e morte celular com elevada potência. Apesar da potência sobre a MCF-7 (carcinoma mamário responsivo ao estrógeno), estudos com estimulação estrogênica revelaram que a atividade da goniotalamina é independente da via hormonal. De qualquer maneira, a goniotalamina racêmica inibiu em 65% a proliferação da MCF-7 implantada em fibras semipermeáveis (hollow fiber) em camundongos. A citometria de fluxo revelou que o racemato de goniotalamina induz apoptose na linhagem tumoral de cólon HT-29 em baixa concentração (10 ?g/mL), com participação das caspases 8 e 9, sugerindo ativação das vias extrínseca e intrínseca. Houve também indução de apoptose na linhagem de melanoma humano SK-MEL-2, mas esse efeito diminuiu quando essa linhagem foi transfectada com Bcl-XL, inibidor apoptótico ligado à mitocôndria, sugerindo ativação principalmente da via apoptótica extrínseca, com ativação da via intrínseca para amplificação do sinal. A goniotalamina em suas três formas comprovou in vivo a atividade apresentada in vitro, pela inibição do desenvolvimento tumoral em modelo de tumor sólido e ascítico de Ehrlich, sem sinais de toxicidade nas doses efetivas. Esses resultados, aliados aos bons rendimentos na rota sintética, favoreceram a continuidade dos estudos com a forma racêmica. Considerando que aproximadamente 25% dos tumores estão relacionados com inflamações crônicas, a goniotalamina racêmica foi avaliada em modelos de edema de pata induzido por carragenina e por diversos mediadores (fosfolipase A2, histamina/serotonina, prostaglandina E2 e bradicinina), apresentando atividade anti-edematogênica em todos os modelos. Corroborando com esses resultados, a goniotalamina apresentou efeito antinociceptivo em modelos de algesia induzidos por processos inflamatórios. A goniotalamina inibiu o desenvolvimento de colite induzida por TNBS e o tratamento oral (30 mg/kg, três vezes por semana/três meses) preveniu o desenvolvimento de inflamação e câncer em animais deficientes em interleucina 10, sem apresentar sinais de toxicidade. Tais dados foram revelados após análise de dano macroscópico e microscópico (histologia) do cólon, havendo diminuição da expressão relativa dos genes para TNF? e MMP9, importantes no processo inflamatório e de invasão e metástase. Além disso, a goniotalamina diminuiu a incidência de metástase pulmonar em modelo de melanoma metastático. Os resultados obtidos e o perfil multi-alvos apresentado pela goniotalamina sugerem sua avaliação em terapias combinatórias para tratamento de processos inflamatórios crônicos e de câncer / Abstract: Carcinogenesis is a long process involving several steps, in which normal cells progressively acquire a neoplastic phenotype. Its origin is favored by genetic factors, exposure to chemical carcinogens, chronic infections and incorporation of mutations into genes involved in regulation of cellular homeostasis. The increasing understanding of tumor biology has provided molecular targets for screening of targeted chemotherapeutic and chemopreventive agents, usually from natural sources or synthesized based on natural products. The improvement of organic chemistry allowed the synthesis of natural products in an economic and refined way, with improvement on their physicochemical properties and availability. This work focuses on goniothalamin, a styryl-lactone naturally found on its enantiomeric form (R), in plants of Goniothalamus (Annonaceae) genus. The (R), (S) and racemic forms were synthesized and led to growth inhibition and cell death in a panel of nine human tumor cell lines, with high potency. Despite the potency against MCF-7 (breast carcinoma responsive to estrogen), estrogen stimulation studies revealed that the goniothalamin activity is independent of the hormone pathway. Anyway, racemic goniothalamin inhibited on 65% the proliferation of MCF-7 implanted into semi permeable fibers (hollow fiber) in mice. Flow cytometry analysis showed that low concentration of racemic goniothalamin (10 ?g/mL) leads colon tumor cell line (HT-29) to apoptosis with involvement of caspase 8 and 9, suggesting the activation of the extrinsic and intrinsic pathways. Apoptosis was also induced in human melanoma cell line SK-MEL-2, but the activity was decreased in SK-MEL-2 transfected with Bcl-XL, a mitochondrial related apoptotic inhibitor, suggesting mainly activation of the extrinsic apoptotic pathway, with activation of the intrinsic pathway to amplify the signal. Goniothalamin on its three forms comproved in vivo the in vitro activity by inhibiting tumor development in models of Ehrlich solid and ascitic tumor in mice, without signals of toxicity at the effective doses. These results, combined with good yields in the synthetic rout favored the racemic form to continue the in vivo studies. Considering that about 25% of tumors are related to chronic inflammation, racemic goniothalamin was evaluated in models of paw edema induced by carrageenan and various mediators (phospholipase A2, histamine/serotonin, prostaglandin E2 and bradykinin) displaying anti-edematogenic activity in all models. Corroborating these results, goniothalamin showed antinociceptive effect in models of algesia induced by inflammatory processes. It also inhibited the development of TNBS-induced colitis and oral treatment (30mg/kg, three times/week, three months) prevented the development of inflammation and cancer in interleukin-10 deficient mice, without signals of toxicity. These data were revealed after macroscopic and microscopic (histology) colon damage analysis, with decreased expression of TNF? and MMP9, which are important in inflammation, invasion and metastasis process. Furthermore, goniothalamin reduced the incidence of lung metastasis in a metastatic melanoma model. These results and the multitarget profile presented by goniothalamin suggest its evaluation in combinatory therapies for treatment of chronic inflammation and cancer / Doutorado / Biologia Celular / Doutor em Biologia Celular e Estrutural

Orientador: Ronaldo Aloise Pilli / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Química / Made available in DSpace on 2018-08-19T04:40:46Z (GMT). No. of bitstreams: 1 Pereira_VanessaCaixeta_M.pdf: 3243379 bytes, checksum: e0935d08b333881437e8413863fa0ff7 (MD5) Previous issue date: 2011 / Resumo: A goniotalamina (9), produto natural isolado de diversas plantas do gênero Goniothalamus, vem se destacando por sua potência em teste de triagem in vitro contra linhagens de células tumorais, bem como pelos resultados positivos em modelos tumorais in vivo. Isso faz com que a goniotalamina sirva como modelo para a síntese e avaliação de novos compostos. Com base nisso, propomos sintetizar e avaliar a atividade citotóxica de duas classes de derivados de 9. A primeira classe são os derivados com substituintes oxigenados na posição 4 da 5,6-di-hidropiran-2-ona. Os derivados alquil e acil (17,24-28) foram obtidos a partir da kavalactona (23) através de reações clássicas de alquilação e acilação em rendimentos que variaram de baixo a excelente. Por sua vez, a kavalactona (23) foi obtida através de uma reação de aldol seguida de lactonização in situ entre o cinamaldeído (10) e acetoacetato de etila (29) em 15% de rendimento. O ensaio de atividade antiproliferativa in vitro mostrou que nenhum dos derivados avaliados foi mais citotóxico que a goniotalamina (9) e que nenhum dos compostos apresentou seletividade para as linhagens avaliadas. Outra classe de compostos avaliada foram os derivados nitrogenados da goniotalamina (9) que apresentam a substituição do anel lactônico por um anel lactâmico. Os derivados lactâmicos 46 a 51 foram sintetizados em 3 etapas e rendimentos globais que variaram de 18 a 60%. A primeira etapa consistiu na formação da imina a partir do respectivo aldeído (10,52-56) seguida de alilação com brometo de alilmagnésio para a formação da amina primária 57. Esta foi convertida em uma amida 58 através da reação com cloreto de acriloíla ou crotonoíla. E por fim, a lactama foi obtida após o tratamento da amida 58 com 6 mol% do catalisador de Grubbs de segunda geração. A avaliação da atividade antiproliferativa in vitro desses derivados mostrou que nenhum deles apresentou citotoxicidade maior que 9. Entretanto, o derivado 49 apresentou seletividade para duas linhagens de células tumorais PC-03 (próstata) e K562 (leucemia) / Abstract: Goniothalamin (9), a natural product isolated from several plants of the Goniothalamus genus, has been highlighted in the literature for presenting good potency against human tumor cell lines in in vitro assays, as well as good results in in vivo models. This makes goniothalamin a promising model for designing new compounds. Herein, we propose the synthesis and the cytotoxic evaluation of two classes of goniothalamin derivatives. The first class comprises derivatives with substituents at position 4 of the 5,6-di-hydropyran-2-one moiety. Alkyl and acyl derivatives (17,24-28) were obtained from kavalactone (23) after well-known alkylation and acylation reaction in bad to good yields. Kavalactone (23) was obtained from cinnamaldehyde (10) and ethyl acetoacetate (29) via an aldol reaction followed by in situ lactonization, in 15% yield. The antiproliferative assay showed that none of these derivatives was more cytotoxic than goniothalamin and none presented selectivity for tumor cell lines. The second class of compounds presents the substitution of the lactone by a lactam moiety. Lactam derivatives 46 - 51 were synthesized in 3 steps and overall yields ranging from 18 to 60%. The first step was the imine formation from the respective aldehyde (10,52-56) followed by allylmagnesium bromide addition to form primary amine 57. This was reacted with acryloyl or crotonoyl chloride to generate amide 58. Finally, the desired lactam was obtained by treating amide 58 with 6 mol% of Grubbs¿ second generation catalyst. The in vitro antiproliferative evaluation showed that 9 was more cytotoxic than the derivatives. However, derivative 49 presented selectivity for two tumor cell lines PC-03 (prostate) and K562 (leukemia) / Mestrado / Quimica Organica / Mestre em Química

Orientador: Ronaldo Aloise Pilli / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Quimica / Made available in DSpace on 2018-08-14T13:10:56Z (GMT). No. of bitstreams: 1 Marquissolo_Cilene_M.pdf: 4919703 bytes, checksum: f834b50c778a98043d8f699e6b2947b9 (MD5) Previous issue date: 2009 / Resumo: Este trabalho tem como objetivo preparar análogos furânicos da goniotalamina 31, 32 e 33 em suas duas formas enantioméricas, utilizando-se a alilação catalítica e assimétrica nas condições de Keck e a reação de metátese de olefinas para fechamento de anel. Estes análogos foram obtidos em bons rendimentos e excelentes razões enantioméricas. Estes novos compostos foram submetidos a testes de atividade antiproliferativa frente a nove linhagens de células tumorais e a bioensaios in vitro contra a forma tripomastigota do Trypanosoma cruzi e contra a forma promastigota de Leishmania major e Leishmania brasiliensis. No que diz respeito à atividade antiproliferativa, exceto para as linhagens de melanoma e cólon, os compostos testados apresentaram maior potência do que o controle positivo, doxorrubicina. Adicionalmente, os compostos 31, 32 e 33 mostraram-se mais ativos que a goniotalamina (1), exceto para as linhagens de mama e ovário. Os bioensaios de citotoxicidade com células não infectadas (LLC-MK2) mostraram que os compostos apresentaram baixa citotoxicidade. Com relação à atividade tripanocida, embora o composto (S)-31 tenha se apresentado como o composto mais ativo, mostrou alta citotoxicidade em células não-infectadas. (S)-32 é o composto que apresenta atividade mais interessante contra T. cruzi, além do menor valor de citotoxicidade e alto índice de segurança. Para a atividade leishmanicida, o análogo (S)-31 apresentou-se como o composto mais ativo para ambas as espécies de Leishmania, mostrando-se cerca de 6,5 vezes mais ativo que o controle positivo para Leishmania brasiliensis e cerca de 14,6 vezes para Leishmania major. Os ensaios de citotoxicidade revelaram valores de concentração superiores aos valores de IC50, indicando baixa toxicidade. Para Leishmania brasiliensis e Leishmania major, (R)-31 e (S)-31 mostraram-se como os compostos mais ativos e apresentaram bons índices de segurança. O análogo 33 não apresentou atividade expressiva em nenhum dos testes realizados indicando que a presença do grupo nitro ligado ao anel furânico é de grande importância para as atividades biológicas avaliadas / Abstract: This work describes the preparation of furan analogues of goniothalamin (compounds 31, 32 and 33) in both enantiomeric forms through the utilization of Keck asymmetric allylation and ring-closing methatesis reaction. These analogues were prepared in good overall yield and excellent enantiomeric ratio. These novel compounds were evaluated as antiproliferative agents against a panel of nine cancer cell lines and in vitro bioassays against the tripomastigote form of Trypanosoma cruzi and promastigote form of Leishmania major and Leishmania brasiliensis. Compounds 31-33 were more potent than the positive control (doxorubicin) in the antiproliferative experiments, except for melanoma and colon cancer cells. Additionally compounds 31-33 were more active than goniothalamin (1), except for breast and ovary cancer cells. Regarding their tripanocidal activity, compound (S)-31 was shown to be very toxic to non-infected cells despite its being highly active. Compound (S)-32 was the most promising one against T. cruzi due to its low toxicity and high insurance level. As to the leishmanicidal activity, the analogue (S)-31 was shown to be the most active for both Leishmania species investigated being 6,5 times more active than the positive control for Leishmania brasiliensis and about 14,6 times for Leishmania major, The citotoxicity assays revealed low toxicity against non-infected cells. For Leishmania brasiliensis and Leishmania major (R)-31 and (S)-31 were shown to be more active one and also displayed the best insurance level. Analogue 33 did not display any significant biological activity in our assays indicating that the nitrofuran moiety is very important for the biological activities investigated / Mestrado / Quimica Organica / Mestre em Química

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第21831号 / 農博第2344号 / 新制||農||1068(附属図書館) / 学位論文||H31||N5203(農学部図書室) / 京都大学大学院農学研究科地域環境科学専攻 / (主査)教授 平井 伸博, 准教授 赤松 美紀, 教授 縄田 栄治 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

Orientadores: Carmen Veríssima Ferreira Halder, Ronaldo Aloise Pilli / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-23T09:59:14Z (GMT). No. of bitstreams: 1 Pelizzaro-Rocha_KarinJuliane_D.pdf: 18909118 bytes, checksum: fa9cec97e3d81e8fb670dab95bc1bc4d (MD5) Previous issue date: 2013 / Resumo: O resumo poderá ser visualizado no texto completo da tese digital quando liberada / Abstract: The abstract is available with the full electronic document when available / Doutorado / Bioquimica / Doutora em Biologia Funcional e Molecular