Identificação do alvo molecular das 2(quinolin-4-ilóxi) acetamidas como candidatos a fármacos para o tratamento da tuberculose
Autor(a) principal: | |
---|---|
Data de Publicação: | 2017 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da PUC_RS |
Texto Completo: | http://tede2.pucrs.br/tede2/handle/tede/7359 |
Resumo: | The high incidence of tuberculosis is a great concern worldwide. Different strategies are being developed by the World Health Organization to fight tuberculosis. Amongst the three pillars that are part of the End TB Strategy, we can highlight the intensive research and innovation pillar. In this extent, the development of new drugs for tuberculosis treatment is a field that is gaining importance. The 2(quinolin-4-yloxy) acetamides are molecules that showed promising bactericidal effects in Mycobacterium tuberculosis, which motivated us to continue studying to improve their mycobactericidal activity and also perform their chemical and biological characterization. In order to continue the 2(quinolin-4-yloxy) acetamides derivatives development, the target identification of these molecules is a keystone and is also the focus of this study. Initially, it was hypothesized that DNA gyrase was their molecular target due to the great chemical similarities between the 2(quinolin-4-yloxy) acetamides and the fluoroquinolones. Despite our results that the 2(quinolin-4-yloxy) acetamides have diminished effects in ofloxacin resistant clinical isolates, the results obtained with the gyrA point mutant and with DNA gyrase protein revealed that this enzyme is not the molecular target of these compounds. The new target identification strategy involved the selection of spontaneous mutants for our lead compound 12L, characterization of this mutant strain against other 2(quinolin-4-yloxy) acetamides derivates and whole genome sequencing. Whole genome sequencing data allowed the identification of a single mutation (T313A) in the QcrB protein, which is the B subunit of cytochrome bc1 complex. This mutation was confirmed by Sanger sequencing and molecular docking results confirmed the importance of this residue for proteindrug interaction. The cytochrome bc1 complex is involved in the electron transport of the bacilli’s respiratory chain, and therefore it appears to be an interesting target, especially to treat the latent form of the disease. We hope that this work contributes to the 2(quinolin-4-yloxy) acetamides development for tuberculosis treatment. |
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Santos, Diógenes Santiago187.292.588-04http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4721461Z7Villela, Anne Drumondhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4126479J1011.970.930-95http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4496890T6Subtil, Fernanda Teixeira2017-06-23T14:56:57Z2017-03-03http://tede2.pucrs.br/tede2/handle/tede/7359The high incidence of tuberculosis is a great concern worldwide. Different strategies are being developed by the World Health Organization to fight tuberculosis. Amongst the three pillars that are part of the End TB Strategy, we can highlight the intensive research and innovation pillar. In this extent, the development of new drugs for tuberculosis treatment is a field that is gaining importance. The 2(quinolin-4-yloxy) acetamides are molecules that showed promising bactericidal effects in Mycobacterium tuberculosis, which motivated us to continue studying to improve their mycobactericidal activity and also perform their chemical and biological characterization. In order to continue the 2(quinolin-4-yloxy) acetamides derivatives development, the target identification of these molecules is a keystone and is also the focus of this study. Initially, it was hypothesized that DNA gyrase was their molecular target due to the great chemical similarities between the 2(quinolin-4-yloxy) acetamides and the fluoroquinolones. Despite our results that the 2(quinolin-4-yloxy) acetamides have diminished effects in ofloxacin resistant clinical isolates, the results obtained with the gyrA point mutant and with DNA gyrase protein revealed that this enzyme is not the molecular target of these compounds. The new target identification strategy involved the selection of spontaneous mutants for our lead compound 12L, characterization of this mutant strain against other 2(quinolin-4-yloxy) acetamides derivates and whole genome sequencing. Whole genome sequencing data allowed the identification of a single mutation (T313A) in the QcrB protein, which is the B subunit of cytochrome bc1 complex. This mutation was confirmed by Sanger sequencing and molecular docking results confirmed the importance of this residue for proteindrug interaction. The cytochrome bc1 complex is involved in the electron transport of the bacilli’s respiratory chain, and therefore it appears to be an interesting target, especially to treat the latent form of the disease. We hope that this work contributes to the 2(quinolin-4-yloxy) acetamides development for tuberculosis treatment.A alta incidência da tuberculose, em âmbito mundial, é de grande preocupação. Para combater esta doença, diferentes estratégias vêm sendo desenvolvidas pela Organização Mundial da Saúde (OMS). Dentre os pilares que compõem a End TB strategy, podemos destacar a pesquisa intensiva e a inovação. Neste âmbito, o desenvolvimento de novos fármacos para tuberculose vem ganhando destaque. As 2(quinolin-4-ilóxi) acetamidas são moléculas que demonstraram resultados bactericidas promissores frente ao Mycobacterium tuberculosis, o que nos motivou a realizar novos estudos para melhorar a atividade e realizar a caracterização química e biológica destas moléculas. A fim de continuar o desenvolvimento da série quinolínica, a identificação do seu alvo molecular foi o foco deste trabalho. Inicialmente, levantou-se a hipótese de que a DNA girase seria o alvo molecular, uma vez que as 2(quinolin- 5-ilóxi) acetamidas possuem grande similaridade estrutural com as fluoroquinolonas. Apesar das 2(quinolin-4-ilóxi) acetamidas terem apresentado menor atividade frente a isolados clínicos resistentes a ofloxacino, os resultados de atividade obtidos frente a uma mutante pontual de gyrA e frente à proteína indicam que a DNA girase não é o alvo destas moléculas. A nova estratégia para identificação de alvo envolveu a seleção de mutantes espontâneos para o composto líder 12L, caracterização desta cepa frente aos demais compostos da série e sequencimento total do genoma. Este permitiu a identificação de uma única mutação (T313A) localizada na proteína QcrB, que é a subunidade B do complexo citocromo bc1. Esta mutação foi confirmada por sequenciamento de Sanger e o docking molecular aferiu a importância deste resíduo na interação proteína-composto. O complexo citocromo bc1 está envolvido no transporte de elétrons na cadeia respiratória do bacilo, e por isso é um alvo molecular interessante, principalmente para combater a forma latente da doença. Esperamos que este trabalho contribua no processo de desenvolvimento das 2(quinolin-4-ilóxi) acetamidas para o tratamento da tuberculose.Submitted by Caroline Xavier (caroline.xavier@pucrs.br) on 2017-06-23T14:56:57Z No. of bitstreams: 1 DIS_FERNANDA_TEIXEIRA_SUBTIL_COMPLETO.pdf: 1495988 bytes, checksum: 70f1cb1dccabfa7c302effb51db76e03 (MD5)Made available in DSpace on 2017-06-23T14:56:57Z (GMT). No. of bitstreams: 1 DIS_FERNANDA_TEIXEIRA_SUBTIL_COMPLETO.pdf: 1495988 bytes, checksum: 70f1cb1dccabfa7c302effb51db76e03 (MD5) Previous issue date: 2017-03-03Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESapplication/pdfhttp://tede2.pucrs.br:80/tede2/retrieve/168717/DIS_FERNANDA_TEIXEIRA_SUBTIL_COMPLETO.pdf.jpgporPontifícia Universidade Católica do Rio Grande do SulPrograma de Pós-Graduação em Biologia Celular e MolecularPUCRSBrasilFaculdade de BiociênciasTuberculoseDesenvolvimento de FármacosIdentificação de AlvoCIENCIAS BIOLOGICAS::BIOLOGIA GERALIdentificação do alvo molecular das 2(quinolin-4-ilóxi) acetamidas como candidatos a fármacos para o tratamento da tuberculoseinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis819824693009663736060060060060036528317262667714-16345593859312446972075167498588264571info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da PUC_RSinstname:Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS)instacron:PUC_RSTHUMBNAILDIS_FERNANDA_TEIXEIRA_SUBTIL_COMPLETO.pdf.jpgDIS_FERNANDA_TEIXEIRA_SUBTIL_COMPLETO.pdf.jpgimage/jpeg3428http://tede2.pucrs.br/tede2/bitstream/tede/7359/5/DIS_FERNANDA_TEIXEIRA_SUBTIL_COMPLETO.pdf.jpgd26f3dd5e384c7322c3017284aa3da76MD55TEXTDIS_FERNANDA_TEIXEIRA_SUBTIL_COMPLETO.pdf.txtDIS_FERNANDA_TEIXEIRA_SUBTIL_COMPLETO.pdf.txttext/plain122644http://tede2.pucrs.br/tede2/bitstream/tede/7359/4/DIS_FERNANDA_TEIXEIRA_SUBTIL_COMPLETO.pdf.txt3e42db97089a5331ef0ba5f95786bc0eMD54LICENSElicense.txtlicense.txttext/plain; charset=utf-8590http://tede2.pucrs.br/tede2/bitstream/tede/7359/3/license.txt220e11f2d3ba5354f917c7035aadef24MD53ORIGINALDIS_FERNANDA_TEIXEIRA_SUBTIL_COMPLETO.pdfDIS_FERNANDA_TEIXEIRA_SUBTIL_COMPLETO.pdfapplication/pdf1495988http://tede2.pucrs.br/tede2/bitstream/tede/7359/2/DIS_FERNANDA_TEIXEIRA_SUBTIL_COMPLETO.pdf70f1cb1dccabfa7c302effb51db76e03MD52tede/73592017-06-23 12:00:52.609oai:tede2.pucrs.br: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Biblioteca Digital de Teses e Dissertaçõeshttp://tede2.pucrs.br/tede2/PRIhttps://tede2.pucrs.br/oai/requestbiblioteca.central@pucrs.br||opendoar:2017-06-23T15:00:52Biblioteca Digital de Teses e Dissertações da PUC_RS - Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS)false |
dc.title.por.fl_str_mv |
Identificação do alvo molecular das 2(quinolin-4-ilóxi) acetamidas como candidatos a fármacos para o tratamento da tuberculose |
title |
Identificação do alvo molecular das 2(quinolin-4-ilóxi) acetamidas como candidatos a fármacos para o tratamento da tuberculose |
spellingShingle |
Identificação do alvo molecular das 2(quinolin-4-ilóxi) acetamidas como candidatos a fármacos para o tratamento da tuberculose Subtil, Fernanda Teixeira Tuberculose Desenvolvimento de Fármacos Identificação de Alvo CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
title_short |
Identificação do alvo molecular das 2(quinolin-4-ilóxi) acetamidas como candidatos a fármacos para o tratamento da tuberculose |
title_full |
Identificação do alvo molecular das 2(quinolin-4-ilóxi) acetamidas como candidatos a fármacos para o tratamento da tuberculose |
title_fullStr |
Identificação do alvo molecular das 2(quinolin-4-ilóxi) acetamidas como candidatos a fármacos para o tratamento da tuberculose |
title_full_unstemmed |
Identificação do alvo molecular das 2(quinolin-4-ilóxi) acetamidas como candidatos a fármacos para o tratamento da tuberculose |
title_sort |
Identificação do alvo molecular das 2(quinolin-4-ilóxi) acetamidas como candidatos a fármacos para o tratamento da tuberculose |
author |
Subtil, Fernanda Teixeira |
author_facet |
Subtil, Fernanda Teixeira |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Santos, Diógenes Santiago |
dc.contributor.advisor1ID.fl_str_mv |
187.292.588-04 |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4721461Z7 |
dc.contributor.advisor-co1.fl_str_mv |
Villela, Anne Drumond |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4126479J1 |
dc.contributor.authorID.fl_str_mv |
011.970.930-95 |
dc.contributor.authorLattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4496890T6 |
dc.contributor.author.fl_str_mv |
Subtil, Fernanda Teixeira |
contributor_str_mv |
Santos, Diógenes Santiago Villela, Anne Drumond |
dc.subject.por.fl_str_mv |
Tuberculose Desenvolvimento de Fármacos Identificação de Alvo |
topic |
Tuberculose Desenvolvimento de Fármacos Identificação de Alvo CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
description |
The high incidence of tuberculosis is a great concern worldwide. Different strategies are being developed by the World Health Organization to fight tuberculosis. Amongst the three pillars that are part of the End TB Strategy, we can highlight the intensive research and innovation pillar. In this extent, the development of new drugs for tuberculosis treatment is a field that is gaining importance. The 2(quinolin-4-yloxy) acetamides are molecules that showed promising bactericidal effects in Mycobacterium tuberculosis, which motivated us to continue studying to improve their mycobactericidal activity and also perform their chemical and biological characterization. In order to continue the 2(quinolin-4-yloxy) acetamides derivatives development, the target identification of these molecules is a keystone and is also the focus of this study. Initially, it was hypothesized that DNA gyrase was their molecular target due to the great chemical similarities between the 2(quinolin-4-yloxy) acetamides and the fluoroquinolones. Despite our results that the 2(quinolin-4-yloxy) acetamides have diminished effects in ofloxacin resistant clinical isolates, the results obtained with the gyrA point mutant and with DNA gyrase protein revealed that this enzyme is not the molecular target of these compounds. The new target identification strategy involved the selection of spontaneous mutants for our lead compound 12L, characterization of this mutant strain against other 2(quinolin-4-yloxy) acetamides derivates and whole genome sequencing. Whole genome sequencing data allowed the identification of a single mutation (T313A) in the QcrB protein, which is the B subunit of cytochrome bc1 complex. This mutation was confirmed by Sanger sequencing and molecular docking results confirmed the importance of this residue for proteindrug interaction. The cytochrome bc1 complex is involved in the electron transport of the bacilli’s respiratory chain, and therefore it appears to be an interesting target, especially to treat the latent form of the disease. We hope that this work contributes to the 2(quinolin-4-yloxy) acetamides development for tuberculosis treatment. |
publishDate |
2017 |
dc.date.accessioned.fl_str_mv |
2017-06-23T14:56:57Z |
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2017-03-03 |
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http://tede2.pucrs.br/tede2/handle/tede/7359 |
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por |
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por |
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8198246930096637360 |
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600 600 600 600 |
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36528317262667714 |
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-1634559385931244697 |
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Pontifícia Universidade Católica do Rio Grande do Sul |
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Programa de Pós-Graduação em Biologia Celular e Molecular |
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PUCRS |
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Brasil |
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Faculdade de Biociências |
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