Avaliação da eficácia e segurança de produtos naturais e sintéticos ativos sobre o Trypanosoma cruzi
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da UERJ |
Texto Completo: | http://www.bdtd.uerj.br/handle/1/20343 |
Resumo: | More than 110 years after the discovery of Chagas Disease (CD), only two nitro-heterocyclic derivatives, associated with partial efficacy and adverse effects, are available for the treatment of approximately 7 million infected people, mainly in Latin America. Considering the importance of bioprospection and rational drug design in the search for more selective therapeutic alternatives, this work investigated, using in silico and in vitro methods, the efficacy and safety of natural and synthetic products with potential application in the control of the parasitic infection by Trypanosoma cruzi. The natural products consisted of a hydromethanolic extract of the leaves of Plinia cauliflora (HME), known as jabuticabeira, and its most abundant phenolic constituents, and the synthetic ones were two new 3-nitro-1,2,4-triazole derivatives, PCBN 009/17 and PCBN 012/17. The metabolomic characterization of the extract was performed by ultra-efficiency liquid chromatography coupled with mass spectrometry. Preliminary in silico analyzes of pharmacokinetic and toxicity (ADMET) properties were conducted on the LAZAR, pkCSM and SwissADME platforms. Subsequently, mutagenicity and genotoxicity were investigated by Ames test and micronucleus assay, in parallel with the determination of cytotoxicity in HepG2 and F C3H liver cells by WST-1 and LDH assays. The in vitro trypanocidal activity was evaluated on trypomastigote forms of the Y strain of T. cruzi. A total of 14 phenolic compounds were identified in HME, mostly flavonoids. In silico predictions for the most abundant compounds, (+)-catechin, digallic acid, and hesperidin, indicated a moderately viable druglikeness profile with low toxicity. In in vitro assays, HME did not produce point mutations in the prokaryotic model, nor an increase in the frequency of micronuclei. However, it exhibited cytostatic effects, reducing nuclear division rates of HepG2. In addition, HME showed time- and dose-dependent cytotoxicity in both strains, promoting cell death through cell membrane damage and more evidently through mitochondrial dysfunction, with F C3H being the most sensitive line. At low concentrations, HME inhibited blood trypomastigotes, generating dose-dependent responses after 2 and 24 hours of exposure. In silico analyzes of the nitrotriazole derivatives indicated structural toxicity alerts for both molecules, which were predicted as hepatotoxic and as substrates of CYP450 enzymes related to drug-drug interactions. Furthermore, the derivatives were predicted to be mutagenic, in agreement with in vitro results, in which they induced base-pair substitution and frameshift mutations. However, the mutagenicity and function of metabolic activation were influenced by the type of substituent, since PCBN 012/17 was mutagenic only at the highest concentration evaluated. The derivatives mainly induced cytotoxicity associated with cell membrane damage, at concentrations higher than their previously described trypanocidal concentrations. In summary, despite the occasional evidence of toxicity, the natural and synthetic products evaluated in this study, especially PCBN 012/17, can be used in the development of more accessible and selective therapeutic alternatives for CD |
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Felzenszwalb, Israelhttp://lattes.cnpq.br/8132847165466920Oliveira, Carlos Fernando Araujo Lima dehttp://lattes.cnpq.br/2590242493425386Coelho, Marsen Garcia Pintohttp://lattes.cnpq.br/4468352500732645Branco, Frederico Silva Castelohttp://lattes.cnpq.br/5370920209323551Mello, Francisco do Vale Chaves ehttp://lattes.cnpq.br/3809738780445226http://lattes.cnpq.br/5229759225844354Galvão, Bárbara Verena Diascba@uerj.br2023-09-22T18:39:36Z2024-01-012022-01-25GALVÃO, Bárbara Verena Dias. Avaliação da eficácia e segurança de produtos naturais e sintéticos ativos sobre o Trypanosoma cruzi. 2022. 118 f. Dissertação (Mestrado em Biociências) – Instituto de Biologia Roberto Alcântara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, 2022.http://www.bdtd.uerj.br/handle/1/20343More than 110 years after the discovery of Chagas Disease (CD), only two nitro-heterocyclic derivatives, associated with partial efficacy and adverse effects, are available for the treatment of approximately 7 million infected people, mainly in Latin America. Considering the importance of bioprospection and rational drug design in the search for more selective therapeutic alternatives, this work investigated, using in silico and in vitro methods, the efficacy and safety of natural and synthetic products with potential application in the control of the parasitic infection by Trypanosoma cruzi. The natural products consisted of a hydromethanolic extract of the leaves of Plinia cauliflora (HME), known as jabuticabeira, and its most abundant phenolic constituents, and the synthetic ones were two new 3-nitro-1,2,4-triazole derivatives, PCBN 009/17 and PCBN 012/17. The metabolomic characterization of the extract was performed by ultra-efficiency liquid chromatography coupled with mass spectrometry. Preliminary in silico analyzes of pharmacokinetic and toxicity (ADMET) properties were conducted on the LAZAR, pkCSM and SwissADME platforms. Subsequently, mutagenicity and genotoxicity were investigated by Ames test and micronucleus assay, in parallel with the determination of cytotoxicity in HepG2 and F C3H liver cells by WST-1 and LDH assays. The in vitro trypanocidal activity was evaluated on trypomastigote forms of the Y strain of T. cruzi. A total of 14 phenolic compounds were identified in HME, mostly flavonoids. In silico predictions for the most abundant compounds, (+)-catechin, digallic acid, and hesperidin, indicated a moderately viable druglikeness profile with low toxicity. In in vitro assays, HME did not produce point mutations in the prokaryotic model, nor an increase in the frequency of micronuclei. However, it exhibited cytostatic effects, reducing nuclear division rates of HepG2. In addition, HME showed time- and dose-dependent cytotoxicity in both strains, promoting cell death through cell membrane damage and more evidently through mitochondrial dysfunction, with F C3H being the most sensitive line. At low concentrations, HME inhibited blood trypomastigotes, generating dose-dependent responses after 2 and 24 hours of exposure. In silico analyzes of the nitrotriazole derivatives indicated structural toxicity alerts for both molecules, which were predicted as hepatotoxic and as substrates of CYP450 enzymes related to drug-drug interactions. Furthermore, the derivatives were predicted to be mutagenic, in agreement with in vitro results, in which they induced base-pair substitution and frameshift mutations. However, the mutagenicity and function of metabolic activation were influenced by the type of substituent, since PCBN 012/17 was mutagenic only at the highest concentration evaluated. The derivatives mainly induced cytotoxicity associated with cell membrane damage, at concentrations higher than their previously described trypanocidal concentrations. In summary, despite the occasional evidence of toxicity, the natural and synthetic products evaluated in this study, especially PCBN 012/17, can be used in the development of more accessible and selective therapeutic alternatives for CDMais de 110 anos após a descoberta da Doença de Chagas (DC), apenas dois derivados nitro-heterocíclicos, associados a eficácia parcial e efeitos adversos, estão disponíveis para o tratamento de cerca de 7 milhões de infectados, sobretudo na América Latina. Considerando a importância da bioprospecção e do desenvolvimento racional de fármacos na busca por novas alternativas terapêuticas mais seletivas, este trabalho investigou, através de métodos in silico e in vitro, a eficácia e segurança de produtos naturais e de síntese com potencial aplicação no controle da infeção parasitária por Trypanosoma cruzi. Os produtos naturais consistiram em um extrato hidrometanólico das folhas de Plinia cauliflora (HME), conhecida como jabuticabeira, e seus constituintes fenólicos mais abundantes, e os sintéticos, foram dois novos derivados 3-nitro-1,2,4-triazólicos, PCBN 009/17 e PCBN 012/17. A caracterização metabolômica do extrato foi realizada por cromatografia líquida de ultra-eficiência acoplada a espectrometria de massas. Análises in silico preliminares de propriedades farmacocinéticas e de toxicidade (ADMET) foram conduzidas nas plataformas LAZAR, pkCSM e SwissADME. Posteriormente, a mutagenicidade e genotoxicidade foi investigada pelo Teste de Ames e ensaio do micronúcleo, em paralelo a determinação de citotoxicidade em células hepáticas HepG2 e F C3H pelos ensaios WST-1 e LDH. A atividade tripanocida in vitro foi avaliada sobre formas tripomastigotas da cepa Y de T. cruzi. Um total de 14 compostos fenólicos foram identificados em HME, majoritariamente flavonoides. As predições in silico para os compostos mais abundantes, (+)- catequina, ácido digálico e hesperidina, indicam um perfil de druglikeness moderadamente viável e de baixa toxicidade. Nos ensaios in vitro, HME não produziu mutações pontuais no modelo procarioto, nem aumento da frequência de micronúcleos. Contudo, exibiu efeitos citostáticos, reduzindo índices de divisão nuclear de HepG2. Complementarmente, HME apresentou citotoxicidade tempo e dose-dependente em ambas as linhagens, promovendo morte celular através de danos à membrana celular e mais evidentemente por disfunção mitocondrial, sendo F C3H, a linhagem mais sensível. Em baixas concentrações, HME inibiu tripomastigotas sanguíneos, gerando respostas dose-dependentes após 2 e 24 horas de exposição. Em continuidade, as análises in silico dos derivados nitrotriazólicos, indicaram alertas estruturais de toxicidade para ambas as moléculas, que foram preditas como hepatotóxicas e como substratos das enzimas CYP450 relacionadas às interações medicamentosas. Além disso, os derivados foram preditos como mutagênicos, em concordância com resultados in vitro, em que induziram mutações por substituição de pares de bases e deslocamento do quadro de leitura. Entretanto, a mutagenicidade e função da ativação metabólica foram influenciadas pelo tipo de substituinte, de forma que o PCBN 012/17 foi mutagênico apenas na maior concentração avaliada. Os derivados induziram principalmente citotoxicidade associada a danos à membrana celular, em concentrações superiores às suas concentrações tripanocidas descritas anteriormente. Em síntese, apesar das evidências pontuais de toxicidade, os produtos naturais e de síntese avaliados no presente estudo, especialmente PCBN 012/17, podem ser empregados no desenvolvimento de alternativas terapêuticas mais acessíveis e seletivas para DCSubmitted by Felipe CB/A (felipebibliotecario@gmail.com) on 2023-09-22T18:39:36Z No. of bitstreams: 2 Dissertação - Barbara Verena Dias Galvão - Completa - 2022.pdf: 3112191 bytes, checksum: 6f3e4144810afbe686355415e3f22d1c (MD5) Dissertação - Barbara Verena Dias Galvão - Parcial - 2022.pdf: 692239 bytes, checksum: 7d6c9098cdd2651567da0afa4b23de45 (MD5)Made available in DSpace on 2023-09-22T18:39:36Z (GMT). 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dc.title.por.fl_str_mv |
Avaliação da eficácia e segurança de produtos naturais e sintéticos ativos sobre o Trypanosoma cruzi |
dc.title.alternative.eng.fl_str_mv |
Efficacy and safety evaluation of natural and synthetic products active on Trypanosoma cruzi |
title |
Avaliação da eficácia e segurança de produtos naturais e sintéticos ativos sobre o Trypanosoma cruzi |
spellingShingle |
Avaliação da eficácia e segurança de produtos naturais e sintéticos ativos sobre o Trypanosoma cruzi Galvão, Bárbara Verena Dias Nitrotriazoles Plinia cauliflora Chagas Disease Trypanosoma cruzi Mutagenicity Cytotoxicity Nitrotriazóis Plinia cauliflora Doença de Chagas Trypanosoma cruzi Mutagenicidade Citotoxicidade Produtos naturais CIENCIAS BIOLOGICAS::GENETICA |
title_short |
Avaliação da eficácia e segurança de produtos naturais e sintéticos ativos sobre o Trypanosoma cruzi |
title_full |
Avaliação da eficácia e segurança de produtos naturais e sintéticos ativos sobre o Trypanosoma cruzi |
title_fullStr |
Avaliação da eficácia e segurança de produtos naturais e sintéticos ativos sobre o Trypanosoma cruzi |
title_full_unstemmed |
Avaliação da eficácia e segurança de produtos naturais e sintéticos ativos sobre o Trypanosoma cruzi |
title_sort |
Avaliação da eficácia e segurança de produtos naturais e sintéticos ativos sobre o Trypanosoma cruzi |
author |
Galvão, Bárbara Verena Dias |
author_facet |
Galvão, Bárbara Verena Dias cba@uerj.br |
author_role |
author |
author2 |
cba@uerj.br |
author2_role |
author |
dc.contributor.advisor1.fl_str_mv |
Felzenszwalb, Israel |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/8132847165466920 |
dc.contributor.advisor-co1.fl_str_mv |
Oliveira, Carlos Fernando Araujo Lima de |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://lattes.cnpq.br/2590242493425386 |
dc.contributor.referee1.fl_str_mv |
Coelho, Marsen Garcia Pinto |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/4468352500732645 |
dc.contributor.referee2.fl_str_mv |
Branco, Frederico Silva Castelo |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/5370920209323551 |
dc.contributor.referee3.fl_str_mv |
Mello, Francisco do Vale Chaves e |
dc.contributor.referee3Lattes.fl_str_mv |
http://lattes.cnpq.br/3809738780445226 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/5229759225844354 |
dc.contributor.author.fl_str_mv |
Galvão, Bárbara Verena Dias cba@uerj.br |
contributor_str_mv |
Felzenszwalb, Israel Oliveira, Carlos Fernando Araujo Lima de Coelho, Marsen Garcia Pinto Branco, Frederico Silva Castelo Mello, Francisco do Vale Chaves e |
dc.subject.eng.fl_str_mv |
Nitrotriazoles Plinia cauliflora Chagas Disease Trypanosoma cruzi Mutagenicity Cytotoxicity |
topic |
Nitrotriazoles Plinia cauliflora Chagas Disease Trypanosoma cruzi Mutagenicity Cytotoxicity Nitrotriazóis Plinia cauliflora Doença de Chagas Trypanosoma cruzi Mutagenicidade Citotoxicidade Produtos naturais CIENCIAS BIOLOGICAS::GENETICA |
dc.subject.por.fl_str_mv |
Nitrotriazóis Plinia cauliflora Doença de Chagas Trypanosoma cruzi Mutagenicidade Citotoxicidade Produtos naturais |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS::GENETICA |
description |
More than 110 years after the discovery of Chagas Disease (CD), only two nitro-heterocyclic derivatives, associated with partial efficacy and adverse effects, are available for the treatment of approximately 7 million infected people, mainly in Latin America. Considering the importance of bioprospection and rational drug design in the search for more selective therapeutic alternatives, this work investigated, using in silico and in vitro methods, the efficacy and safety of natural and synthetic products with potential application in the control of the parasitic infection by Trypanosoma cruzi. The natural products consisted of a hydromethanolic extract of the leaves of Plinia cauliflora (HME), known as jabuticabeira, and its most abundant phenolic constituents, and the synthetic ones were two new 3-nitro-1,2,4-triazole derivatives, PCBN 009/17 and PCBN 012/17. The metabolomic characterization of the extract was performed by ultra-efficiency liquid chromatography coupled with mass spectrometry. Preliminary in silico analyzes of pharmacokinetic and toxicity (ADMET) properties were conducted on the LAZAR, pkCSM and SwissADME platforms. Subsequently, mutagenicity and genotoxicity were investigated by Ames test and micronucleus assay, in parallel with the determination of cytotoxicity in HepG2 and F C3H liver cells by WST-1 and LDH assays. The in vitro trypanocidal activity was evaluated on trypomastigote forms of the Y strain of T. cruzi. A total of 14 phenolic compounds were identified in HME, mostly flavonoids. In silico predictions for the most abundant compounds, (+)-catechin, digallic acid, and hesperidin, indicated a moderately viable druglikeness profile with low toxicity. In in vitro assays, HME did not produce point mutations in the prokaryotic model, nor an increase in the frequency of micronuclei. However, it exhibited cytostatic effects, reducing nuclear division rates of HepG2. In addition, HME showed time- and dose-dependent cytotoxicity in both strains, promoting cell death through cell membrane damage and more evidently through mitochondrial dysfunction, with F C3H being the most sensitive line. At low concentrations, HME inhibited blood trypomastigotes, generating dose-dependent responses after 2 and 24 hours of exposure. In silico analyzes of the nitrotriazole derivatives indicated structural toxicity alerts for both molecules, which were predicted as hepatotoxic and as substrates of CYP450 enzymes related to drug-drug interactions. Furthermore, the derivatives were predicted to be mutagenic, in agreement with in vitro results, in which they induced base-pair substitution and frameshift mutations. However, the mutagenicity and function of metabolic activation were influenced by the type of substituent, since PCBN 012/17 was mutagenic only at the highest concentration evaluated. The derivatives mainly induced cytotoxicity associated with cell membrane damage, at concentrations higher than their previously described trypanocidal concentrations. In summary, despite the occasional evidence of toxicity, the natural and synthetic products evaluated in this study, especially PCBN 012/17, can be used in the development of more accessible and selective therapeutic alternatives for CD |
publishDate |
2022 |
dc.date.issued.fl_str_mv |
2022-01-25 |
dc.date.accessioned.fl_str_mv |
2023-09-22T18:39:36Z |
dc.date.available.fl_str_mv |
2024-01-01 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
GALVÃO, Bárbara Verena Dias. Avaliação da eficácia e segurança de produtos naturais e sintéticos ativos sobre o Trypanosoma cruzi. 2022. 118 f. Dissertação (Mestrado em Biociências) – Instituto de Biologia Roberto Alcântara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, 2022. |
dc.identifier.uri.fl_str_mv |
http://www.bdtd.uerj.br/handle/1/20343 |
identifier_str_mv |
GALVÃO, Bárbara Verena Dias. Avaliação da eficácia e segurança de produtos naturais e sintéticos ativos sobre o Trypanosoma cruzi. 2022. 118 f. Dissertação (Mestrado em Biociências) – Instituto de Biologia Roberto Alcântara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, 2022. |
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http://www.bdtd.uerj.br/handle/1/20343 |
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por |
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por |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
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Universidade do Estado do Rio de Janeiro |
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Programa de Pós-Graduação em Biociências |
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UERJ |
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Brasil |
dc.publisher.department.fl_str_mv |
Centro Biomédico::Instituto de Biologia Roberto Alcantara Gomes |
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Universidade do Estado do Rio de Janeiro |
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UERJ |
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UERJ |
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Biblioteca Digital de Teses e Dissertações da UERJ |
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Biblioteca Digital de Teses e Dissertações da UERJ |
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Biblioteca Digital de Teses e Dissertações da UERJ - Universidade do Estado do Rio de Janeiro (UERJ) |
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bdtd.suporte@uerj.br |
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1811728739599384576 |