Atividade antifúngica contra Aspergillus e caracterização toxicológica da amida sintética 2- cloro-N-fenilacetamida
Ano de defesa: | 2021 |
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Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | |
Tipo de documento: | Tese |
Tipo de acesso: | Acesso aberto |
Idioma: | por |
Instituição de defesa: |
Universidade Federal da Paraíba
Brasil Farmacologia Programa de Pós-Graduação em Produtos Naturais e Sintéticos Bioativos UFPB |
Programa de Pós-Graduação: |
Não Informado pela instituição
|
Departamento: |
Não Informado pela instituição
|
País: |
Não Informado pela instituição
|
Palavras-chave em Português: | |
Link de acesso: | https://repositorio.ufpb.br/jspui/handle/123456789/22826 |
Resumo: | The Aspergillus genus comprises a group of saprophytic fungi that cause opportunistic infections that mainly affect immunocompromised individuals. The species A. flavus and A. niger are, respectively, the second and third major causes of aspergillosis documented in the literature. Knowing that Aspergillus spp resistance to current therapies has been growing and reducing the possibilities of treatment with available antifungal agents, and that the synthetic amide 2-chloro-N-phenylacetamide has shown antifungal potential against this genus, the aim of this study was to evaluate the effect of synthetic amide 2-chloro-N-phenylacetamide on Aspergillus and to determine its toxicological profile. Therefore, the minimum inhibitory concentration, minimum fungicidal concentration, conidia germination, associations with antifungal agents, elucidation of the probable mechanism of action by assays involving fungal cell wall and membrane, as well as molecular prediction by docking, were evaluated. The pharmacokinetic and toxicological profile was evaluated from in silico assays using Swiss ADMET and OSIRIS software. In addition, cytotoxicity was evaluated in vitro through the hemolysis test, the protective capacity through the osmotic fragility test and the ex vivo genotoxic effects on cells of the oral mucosa. It was observed that 2-chloro-N-phenylacetamide showed antifungal activity against A. flavus strains with minimal inhibitory concentrations between 16 and 256 μg/mL and minimal fungicidal concentrations ranging from 32 to 512 μg/mL. The evaluated compound showed antifungal activity against A. niger strains with MICs and CFMs ranging, respectively, from 32 and 256 μg/mL, and 64 to 1024 μg/mL, thus exhibiting fungicidal activity for all strains used. For both species, 2-chloro-N-phenylacetamide promoted inhibition of conidia germination at all concentrations used, and the association with amphotericin B or voriconazole was antagonistic. Ergosterol binding in the fungal plasma membrane is the likely mechanism of action, together with the possible inhibition of DNA synthesis through inhibition of thymidylate synthase. In the evaluation of the in silico pharmacokinetic and in vitro and ex vivo toxicological profile, it was observed that 2-chloro-N-phenylacetamide presents favorable physicochemical parameters according to Lipinski's rule, presenting good oral bioavailability, absorption through the tract gastrointestinal, ability to cross the bloodbrain barrier, inhibition of CYP1A2 and mutagenic, tumorigenic, and reproductiverelated damage effects. 2-chloro-N-phenylacetamide, has low hemolytic capacity at concentrations of 50, 100 and 500 µg/mL, medium toxicity at a concentration of 1000 µg/mL and shows a protective effect of red blood cells of blood types A and O. The compound promotes few genotoxic changes, including karyolysis and karyorrhexis in cells of the oral mucosa, but at a lower frequency than that observed for the positive control hydrogen peroxide. Therefore, it can be concluded that 2-chloro-Nphenylacetamide has promising antifungal potential with a favorable pharmacokinetic profile for oral administration, and low cytotoxic and genotoxic potential, being a promising candidate for in vivo toxicity studies. |
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Atividade antifúngica contra Aspergillus e caracterização toxicológica da amida sintética 2- cloro-N-fenilacetamidaAspergillusAntifúngicoAcetamidasErgosterolToxicidadeAntifungalAcetamidesToxicityCNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIAThe Aspergillus genus comprises a group of saprophytic fungi that cause opportunistic infections that mainly affect immunocompromised individuals. The species A. flavus and A. niger are, respectively, the second and third major causes of aspergillosis documented in the literature. Knowing that Aspergillus spp resistance to current therapies has been growing and reducing the possibilities of treatment with available antifungal agents, and that the synthetic amide 2-chloro-N-phenylacetamide has shown antifungal potential against this genus, the aim of this study was to evaluate the effect of synthetic amide 2-chloro-N-phenylacetamide on Aspergillus and to determine its toxicological profile. Therefore, the minimum inhibitory concentration, minimum fungicidal concentration, conidia germination, associations with antifungal agents, elucidation of the probable mechanism of action by assays involving fungal cell wall and membrane, as well as molecular prediction by docking, were evaluated. The pharmacokinetic and toxicological profile was evaluated from in silico assays using Swiss ADMET and OSIRIS software. In addition, cytotoxicity was evaluated in vitro through the hemolysis test, the protective capacity through the osmotic fragility test and the ex vivo genotoxic effects on cells of the oral mucosa. It was observed that 2-chloro-N-phenylacetamide showed antifungal activity against A. flavus strains with minimal inhibitory concentrations between 16 and 256 μg/mL and minimal fungicidal concentrations ranging from 32 to 512 μg/mL. The evaluated compound showed antifungal activity against A. niger strains with MICs and CFMs ranging, respectively, from 32 and 256 μg/mL, and 64 to 1024 μg/mL, thus exhibiting fungicidal activity for all strains used. For both species, 2-chloro-N-phenylacetamide promoted inhibition of conidia germination at all concentrations used, and the association with amphotericin B or voriconazole was antagonistic. Ergosterol binding in the fungal plasma membrane is the likely mechanism of action, together with the possible inhibition of DNA synthesis through inhibition of thymidylate synthase. In the evaluation of the in silico pharmacokinetic and in vitro and ex vivo toxicological profile, it was observed that 2-chloro-N-phenylacetamide presents favorable physicochemical parameters according to Lipinski's rule, presenting good oral bioavailability, absorption through the tract gastrointestinal, ability to cross the bloodbrain barrier, inhibition of CYP1A2 and mutagenic, tumorigenic, and reproductiverelated damage effects. 2-chloro-N-phenylacetamide, has low hemolytic capacity at concentrations of 50, 100 and 500 µg/mL, medium toxicity at a concentration of 1000 µg/mL and shows a protective effect of red blood cells of blood types A and O. The compound promotes few genotoxic changes, including karyolysis and karyorrhexis in cells of the oral mucosa, but at a lower frequency than that observed for the positive control hydrogen peroxide. Therefore, it can be concluded that 2-chloro-Nphenylacetamide has promising antifungal potential with a favorable pharmacokinetic profile for oral administration, and low cytotoxic and genotoxic potential, being a promising candidate for in vivo toxicity studies.NenhumaO gênero Aspergillus compreende um grupo de fungos saprófitos, causadores de infecções oportunistas que acometem principalmente indivíduos imunocomprometidos. As espécies A. flavus e A. niger são, respectivamente, a segunda e terceira maiores causas de aspergilose documentada na literatura. Sabendo que a resistência de Aspergillus spp às atuais terapias vêm crescendo e reduzindo as possibilidades de tratamento com os antifúngicos disponíveis, e que a amida sintética 2-cloro-N-fenilacetamida têm mostrado potencial antifúngico contra esse gênero, o objetivo desse trabalho foi avaliar o efeito da amida sintética 2-cloroN-fenilacetamida sobre Aspergillus e determinar seu perfil toxicológico. Para tanto, foi avaliado a concentração inibitória mínima, concentração fungicida mínima, germinação de conídios, associações com antifúngicos, elucidação do provável mecanismo de ação por ensaios que envolvem parede e membrana celular fúngica, bem como predição molécular pelo docking. O perfil farmacocinético e toxicológico foi avaliado a partir de ensaios in silico utilizando os softwares Swiss ADMET e OSIRIS. Além disso, foi avaliado in vitro a citotoxicidade por meio do teste de hemólise, a capacidade protetiva por meio do teste de fragilidade osmótica e os efeitos genotóxicos ex vivo em células da mucosa oral. Foi observado que 2-cloro-Nfenilacetamida apresentou atividade antifúngica contra cepas de A. flavus com concentrações inibitórias mínimas entre 16 e 256 μg/mL e concentrações fungicidas mínimas que variaram de 32 a 512 μg/mL. O composto avaliado apresentou atividade antifúngica contra cepas de A. niger com CIMs e CFMs variando, respectivamente, de 32 e 256 μg/mL, e 64 a 1024 μg/mL, exibindo, portanto, atividade fungicida para todas as cepas utilizadas. Para ambas as espécies, 2-cloroN-fenilacetamida promoveu inibição da germinação dos conídios em todas as concentrações utilizadas, e a associação com à anfotericina B ou ao voriconazol foi antagônica. A ligação ao ergosterol na membrana plasmática fúngica é o provável mecanismo de ação, junto com a possível inibição da síntese de DNA por meio da inibição da timidilato sintase. Na avaliação do perfil farmacocinético in silico e toxicológico in vitro e ex vivo, foi observado que a 2-cloro-N-fenilacetamida apresenta parâmetros físico-quimicos favoráveis de acordo com a regra de Lipinski, apresentando boa biodisponibilidade por via oral, absorção pelo trato gastrointestinal, capacidade de atravessar a barreira hematoencefálica, inibição da CYP1A2 e efeitos mutagênicos, tumorigênicos e danos relacionados à reprodução. 2-cloro-N-fenilacetamida, apresenta baixa capacidade hemolítica nas concentrações de 50, 100 e 500 µg/mL, média toxicidade na concentração de 1000 µg/mL e mostra efeito protetivo das hemácias dos tipos sanguíneos A e O. Além disso, o composto promove poucas alterações genotóxicas, incluindo cariólise e cariorrexe em células da mucosa oral, porém em frequência menor do que o observado para o controle positivo peróxido de hidrogênio. Diante disso, pode-se concluir que 2-cloro-Nfenilacetamida apresenta potencial antifúngico promissor com perfil farmacocinético favorável a administração pela via oral, e baixo potencial citotóxico e genotóxico, sendo um candidato promissor para estudos de toxicidade in vivo.Universidade Federal da ParaíbaBrasilFarmacologiaPrograma de Pós-Graduação em Produtos Naturais e Sintéticos BioativosUFPBCastro, Ricardo Dias dehttp://lattes.cnpq.br/0031529469046003Lima, Edeltrudes de Oliveirahttp://lattes.cnpq.br/9406572870167006Ferreira, Elba dos Santos2022-05-12T16:51:01Z2021-09-102022-05-12T16:51:01Z2021-08-09info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesishttps://repositorio.ufpb.br/jspui/handle/123456789/22826porAttribution-NoDerivs 3.0 Brazilhttp://creativecommons.org/licenses/by-nd/3.0/br/info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UFPBinstname:Universidade Federal da Paraíba (UFPB)instacron:UFPB2022-08-09T14:33:13Zoai:repositorio.ufpb.br:123456789/22826Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufpb.br/PUBhttp://tede.biblioteca.ufpb.br:8080/oai/requestdiretoria@ufpb.br|| diretoria@ufpb.bropendoar:2022-08-09T14:33:13Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB)false |
dc.title.none.fl_str_mv |
Atividade antifúngica contra Aspergillus e caracterização toxicológica da amida sintética 2- cloro-N-fenilacetamida |
title |
Atividade antifúngica contra Aspergillus e caracterização toxicológica da amida sintética 2- cloro-N-fenilacetamida |
spellingShingle |
Atividade antifúngica contra Aspergillus e caracterização toxicológica da amida sintética 2- cloro-N-fenilacetamida Ferreira, Elba dos Santos Aspergillus Antifúngico Acetamidas Ergosterol Toxicidade Antifungal Acetamides Toxicity CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
title_short |
Atividade antifúngica contra Aspergillus e caracterização toxicológica da amida sintética 2- cloro-N-fenilacetamida |
title_full |
Atividade antifúngica contra Aspergillus e caracterização toxicológica da amida sintética 2- cloro-N-fenilacetamida |
title_fullStr |
Atividade antifúngica contra Aspergillus e caracterização toxicológica da amida sintética 2- cloro-N-fenilacetamida |
title_full_unstemmed |
Atividade antifúngica contra Aspergillus e caracterização toxicológica da amida sintética 2- cloro-N-fenilacetamida |
title_sort |
Atividade antifúngica contra Aspergillus e caracterização toxicológica da amida sintética 2- cloro-N-fenilacetamida |
author |
Ferreira, Elba dos Santos |
author_facet |
Ferreira, Elba dos Santos |
author_role |
author |
dc.contributor.none.fl_str_mv |
Castro, Ricardo Dias de http://lattes.cnpq.br/0031529469046003 Lima, Edeltrudes de Oliveira http://lattes.cnpq.br/9406572870167006 |
dc.contributor.author.fl_str_mv |
Ferreira, Elba dos Santos |
dc.subject.por.fl_str_mv |
Aspergillus Antifúngico Acetamidas Ergosterol Toxicidade Antifungal Acetamides Toxicity CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
topic |
Aspergillus Antifúngico Acetamidas Ergosterol Toxicidade Antifungal Acetamides Toxicity CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
description |
The Aspergillus genus comprises a group of saprophytic fungi that cause opportunistic infections that mainly affect immunocompromised individuals. The species A. flavus and A. niger are, respectively, the second and third major causes of aspergillosis documented in the literature. Knowing that Aspergillus spp resistance to current therapies has been growing and reducing the possibilities of treatment with available antifungal agents, and that the synthetic amide 2-chloro-N-phenylacetamide has shown antifungal potential against this genus, the aim of this study was to evaluate the effect of synthetic amide 2-chloro-N-phenylacetamide on Aspergillus and to determine its toxicological profile. Therefore, the minimum inhibitory concentration, minimum fungicidal concentration, conidia germination, associations with antifungal agents, elucidation of the probable mechanism of action by assays involving fungal cell wall and membrane, as well as molecular prediction by docking, were evaluated. The pharmacokinetic and toxicological profile was evaluated from in silico assays using Swiss ADMET and OSIRIS software. In addition, cytotoxicity was evaluated in vitro through the hemolysis test, the protective capacity through the osmotic fragility test and the ex vivo genotoxic effects on cells of the oral mucosa. It was observed that 2-chloro-N-phenylacetamide showed antifungal activity against A. flavus strains with minimal inhibitory concentrations between 16 and 256 μg/mL and minimal fungicidal concentrations ranging from 32 to 512 μg/mL. The evaluated compound showed antifungal activity against A. niger strains with MICs and CFMs ranging, respectively, from 32 and 256 μg/mL, and 64 to 1024 μg/mL, thus exhibiting fungicidal activity for all strains used. For both species, 2-chloro-N-phenylacetamide promoted inhibition of conidia germination at all concentrations used, and the association with amphotericin B or voriconazole was antagonistic. Ergosterol binding in the fungal plasma membrane is the likely mechanism of action, together with the possible inhibition of DNA synthesis through inhibition of thymidylate synthase. In the evaluation of the in silico pharmacokinetic and in vitro and ex vivo toxicological profile, it was observed that 2-chloro-N-phenylacetamide presents favorable physicochemical parameters according to Lipinski's rule, presenting good oral bioavailability, absorption through the tract gastrointestinal, ability to cross the bloodbrain barrier, inhibition of CYP1A2 and mutagenic, tumorigenic, and reproductiverelated damage effects. 2-chloro-N-phenylacetamide, has low hemolytic capacity at concentrations of 50, 100 and 500 µg/mL, medium toxicity at a concentration of 1000 µg/mL and shows a protective effect of red blood cells of blood types A and O. The compound promotes few genotoxic changes, including karyolysis and karyorrhexis in cells of the oral mucosa, but at a lower frequency than that observed for the positive control hydrogen peroxide. Therefore, it can be concluded that 2-chloro-Nphenylacetamide has promising antifungal potential with a favorable pharmacokinetic profile for oral administration, and low cytotoxic and genotoxic potential, being a promising candidate for in vivo toxicity studies. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-09-10 2021-08-09 2022-05-12T16:51:01Z 2022-05-12T16:51:01Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufpb.br/jspui/handle/123456789/22826 |
url |
https://repositorio.ufpb.br/jspui/handle/123456789/22826 |
dc.language.iso.fl_str_mv |
por |
language |
por |
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Attribution-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nd/3.0/br/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nd/3.0/br/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal da Paraíba Brasil Farmacologia Programa de Pós-Graduação em Produtos Naturais e Sintéticos Bioativos UFPB |
publisher.none.fl_str_mv |
Universidade Federal da Paraíba Brasil Farmacologia Programa de Pós-Graduação em Produtos Naturais e Sintéticos Bioativos UFPB |
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reponame:Biblioteca Digital de Teses e Dissertações da UFPB instname:Universidade Federal da Paraíba (UFPB) instacron:UFPB |
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UFPB |
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UFPB |
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Biblioteca Digital de Teses e Dissertações da UFPB |
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Biblioteca Digital de Teses e Dissertações da UFPB |
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Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB) |
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diretoria@ufpb.br|| diretoria@ufpb.br |
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