Avaliação da atividade antifúngica de 2-Cloro-N-fenilacetamida (a1cl) contra espécies de candida de origem clínica
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da UFPB |
Texto Completo: | https://repositorio.ufpb.br/jspui/handle/123456789/29761 |
Resumo: | Candida and mycoses are a growing global public health concern. Candida albicans, Candida tropicalis and Candida parapsilosis are on the list of priority fungal pathogens created by the World Health Organization. Thus, the development of research is planned to generate new drugs in order to combat and prevent the development of resistance. In this sense, prospective studies with reaction intermediates with amide groups and chlorine atoms are a promising strategy. This work aimed to evaluate the antifungal activity of the synthetic molecule 2-Chloro- N-phenylacetamides (A1Cl) against C. albicans, C. tropicalis and C. parapsilosis. The minimum inhibitory concentration (MIC) was determined by microdilution, so that values ranged from 16 to 256 μg/mL for A1Cl and 8 to 512 μg/mL for fluconazole (positive control). The minimum fungicidal concentration was similar to the MIC, for both evaluated compounds, confirming the fungicidal effect of A1Cl. In the model of morphological changes (microculture technique), A1Cl and fluconazole reduced the formation of virulence structures compared to their respective controls, showing a concentration-dependent effect. The presence of sorbitol and exogenous ergosterol did not alter the MIC values, confirming that A1Cl has no effect interfering with the functionality and/or integrity of the cell wall and membrane, respectively. The association study (checkerboard) of A1Cl with fluconazole resulted in an antifungal effect of indifference. In the in vitro antibiofilm effect assay, A1Cl was able to inhibit biofilm formation by more than 45% and 80% (***p<0.001), and was able to promote preformed biofilm rupture by more than 35% and 60 % (***p<0.001), in sub and supra inhibitory concentrations, respectively, when compared to the respective control groups. However, for fluconazole to achieve 50% (***p<0.001) or more inhibition effect, suprainhibitory concentrations were required. Similarly, A1Cl (CIM) was able to inhibit by more than 50% (***p<0.001) the formation of ex vivo biofilms (nail fragments) and for fluconazole to achieve the same antibiofilm effect by more than 50% 4xCIM was required. A1Cl showed binding energy values greater than or close to fluconazole in at least one scoring function, for Dihydrofolate reductase (DHFR), Geranylgeranyltransferase-I (GGTase-I) and Aspartic Protease-2 (SAP-2), so that these enzymes, may be targets susceptible to the antifungal action of A1Cl. The RMSD of the evaluated complexes revealed stability around 2-3ns and maintained its stability above 10 ns. For the RMSF of the complexes, we observed interactions similar to those investigated in docking after dynamic simulations at times of 200 and 600ps and that A1Cl remained in the active site under the influence of solvents and structural flexibility. Given these results, it is possible to infer that A1Cl has an antifungal effect and that this activity is related to antibiofilm mechanisms and binding to DHFR and SAP-2. |
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Avaliação da atividade antifúngica de 2-Cloro-N-fenilacetamida (a1cl) contra espécies de candida de origem clínicaFungicida - Candida2-Cloro-N-FenilacetamidaAntibiofilmesDocking molecularDinâmica molecularFungicide - Candida2-Chloro-N-PhenylacetamideAntibiofilmsMolecular dockingMolecular dinamicCNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIACandida and mycoses are a growing global public health concern. Candida albicans, Candida tropicalis and Candida parapsilosis are on the list of priority fungal pathogens created by the World Health Organization. Thus, the development of research is planned to generate new drugs in order to combat and prevent the development of resistance. In this sense, prospective studies with reaction intermediates with amide groups and chlorine atoms are a promising strategy. This work aimed to evaluate the antifungal activity of the synthetic molecule 2-Chloro- N-phenylacetamides (A1Cl) against C. albicans, C. tropicalis and C. parapsilosis. The minimum inhibitory concentration (MIC) was determined by microdilution, so that values ranged from 16 to 256 μg/mL for A1Cl and 8 to 512 μg/mL for fluconazole (positive control). The minimum fungicidal concentration was similar to the MIC, for both evaluated compounds, confirming the fungicidal effect of A1Cl. In the model of morphological changes (microculture technique), A1Cl and fluconazole reduced the formation of virulence structures compared to their respective controls, showing a concentration-dependent effect. The presence of sorbitol and exogenous ergosterol did not alter the MIC values, confirming that A1Cl has no effect interfering with the functionality and/or integrity of the cell wall and membrane, respectively. The association study (checkerboard) of A1Cl with fluconazole resulted in an antifungal effect of indifference. In the in vitro antibiofilm effect assay, A1Cl was able to inhibit biofilm formation by more than 45% and 80% (***p<0.001), and was able to promote preformed biofilm rupture by more than 35% and 60 % (***p<0.001), in sub and supra inhibitory concentrations, respectively, when compared to the respective control groups. However, for fluconazole to achieve 50% (***p<0.001) or more inhibition effect, suprainhibitory concentrations were required. Similarly, A1Cl (CIM) was able to inhibit by more than 50% (***p<0.001) the formation of ex vivo biofilms (nail fragments) and for fluconazole to achieve the same antibiofilm effect by more than 50% 4xCIM was required. A1Cl showed binding energy values greater than or close to fluconazole in at least one scoring function, for Dihydrofolate reductase (DHFR), Geranylgeranyltransferase-I (GGTase-I) and Aspartic Protease-2 (SAP-2), so that these enzymes, may be targets susceptible to the antifungal action of A1Cl. The RMSD of the evaluated complexes revealed stability around 2-3ns and maintained its stability above 10 ns. For the RMSF of the complexes, we observed interactions similar to those investigated in docking after dynamic simulations at times of 200 and 600ps and that A1Cl remained in the active site under the influence of solvents and structural flexibility. Given these results, it is possible to infer that A1Cl has an antifungal effect and that this activity is related to antibiofilm mechanisms and binding to DHFR and SAP-2.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESCandida e micoses são uma preocupação crescente de saúde pública global. Candida albicans, Candida tropicalis e Candida parapsilosis estão na lista de patógenos fúngicos prioritários criada pela Organização mundial de saúde. Assim, é planejado o desenvolvimento de pesquisas para geração de novos fármacos com intuito de combater e prevenir o desenvolvimento de resistência. Neste sentido, estudos de prospecção com intermediários de reações com grupamentos de amidas e átomos de cloro são uma estatégia promissora. Este trabalho teve como objetivo avaliar a atividade antifúngica da molécula sintética 2-Cloro-N-fenilacetamidas (A1Cl) , frente C. albicans, C. tropicalis e C. parapsilosis. A concentração inibitória mínima (CIM) foi determinada por microdiluição, de modo que os valores variaram de 16 a 256 μg/mL para A1Cl e 8 a 512 μg/mL para fluconazol (controle positivo). A concentração fungicida mínima foi semelhante a CIM, para ambos os compostos avaliados, confirmando efeito fungicida de A1Cl. No modelo de alterações morfológicas (técnica de microcultivo), A1Cl e fluconazol reduziu a formação de estruturas de virulência comparados aos seus respectivos controles, evidenciando um efeito dependente de concentração. A presença de sorbitol e ergosterol exógeno não alterou os valores de CIM, confirmando que A1Cl não exerce efeito inteferindo na funcionalidade e/ou integridade da parede e membrana celular, respectivamente. O estudo de associação (checkerboard) de A1Cl com fluconazol resultou em efeito antifúngico de indiferença. No ensaio de efeito antibiofilme in vitro A1Cl foi capaz de inibir a formação de biofilme em mais de 45% e 80% (***p<0,001), e capaz de promover ruptura de biofilme pré- formado em mais de 35% e 60% (***p<0,001), em concentrações sub e suprainibitórias, respectivamente, quando comparados aos respectivos grupos controle. Todavia, para fluconazol alcançar 50% (***p<0,001) ou mais de efeito de inibição foi necessário concentrações suprainibitórias. De forma semelhante, A1Cl (CIM) foi capaz de inibir em mais de 50% (***p<0,001) a formação de biofilmes em modelo ex vivo (fragmentos de unhas) e para o fluconazol alcançar o mesmo efeito antibiofilme em mais de 50% foi necessário 4xCIM. A1Cl apresentou valores de energia de ligação maiores ou próximos ao fluconazol em pelo menos uma função de pontuação, para Dihidrofolato redutase (DHFR), Geranilgeraniltransferase-I (GGTase-I) e Protease aspártica-2 (SAP-2), de modo que essas enzimas, podem ser alvos susceptíveis à ação antifúngica de A1Cl. O RMSD dos complexos avaliados revelou estabilidade em torno de 2-3ns e manteve sua estabilidade acima de 10 ns. Para o RMSF dos complexos, observamos interações semelhantes às pesquisadas no docking após simulações dinâmicas em tempos de 200 e 600ps e que A1Cl permaneceu no sítio ativo sob a influência de solventes e flexibilidade estrutural. Diante desses resultados é possível inferir que A1Cl apresenta efeito antifúngico e que essa atividade está relacionada com mecanismos antibiofilmes e ligação à DHFR e SAP-2.Universidade Federal da ParaíbaBrasilFarmacologiaPrograma de Pós-Graduação em Produtos Naturais e Sintéticos BioativosUFPBLima, Edeltrudes de Oliveirahttp://lattes.cnpq.br/9406572870167006Lima, Alberto Shellygton2024-03-06T13:01:11Z2023-07-202024-03-06T13:01:11Z2023-03-03info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesishttps://repositorio.ufpb.br/jspui/handle/123456789/29761porAttribution-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:UFPB2024-03-07T06:07:25Zoai:repositorio.ufpb.br:123456789/29761Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufpb.br/PUBhttp://tede.biblioteca.ufpb.br:8080/oai/requestdiretoria@ufpb.br|| diretoria@ufpb.bropendoar:2024-03-07T06:07:25Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB)false |
dc.title.none.fl_str_mv |
Avaliação da atividade antifúngica de 2-Cloro-N-fenilacetamida (a1cl) contra espécies de candida de origem clínica |
title |
Avaliação da atividade antifúngica de 2-Cloro-N-fenilacetamida (a1cl) contra espécies de candida de origem clínica |
spellingShingle |
Avaliação da atividade antifúngica de 2-Cloro-N-fenilacetamida (a1cl) contra espécies de candida de origem clínica Lima, Alberto Shellygton Fungicida - Candida 2-Cloro-N-Fenilacetamida Antibiofilmes Docking molecular Dinâmica molecular Fungicide - Candida 2-Chloro-N-Phenylacetamide Antibiofilms Molecular docking Molecular dinamic CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
title_short |
Avaliação da atividade antifúngica de 2-Cloro-N-fenilacetamida (a1cl) contra espécies de candida de origem clínica |
title_full |
Avaliação da atividade antifúngica de 2-Cloro-N-fenilacetamida (a1cl) contra espécies de candida de origem clínica |
title_fullStr |
Avaliação da atividade antifúngica de 2-Cloro-N-fenilacetamida (a1cl) contra espécies de candida de origem clínica |
title_full_unstemmed |
Avaliação da atividade antifúngica de 2-Cloro-N-fenilacetamida (a1cl) contra espécies de candida de origem clínica |
title_sort |
Avaliação da atividade antifúngica de 2-Cloro-N-fenilacetamida (a1cl) contra espécies de candida de origem clínica |
author |
Lima, Alberto Shellygton |
author_facet |
Lima, Alberto Shellygton |
author_role |
author |
dc.contributor.none.fl_str_mv |
Lima, Edeltrudes de Oliveira http://lattes.cnpq.br/9406572870167006 |
dc.contributor.author.fl_str_mv |
Lima, Alberto Shellygton |
dc.subject.por.fl_str_mv |
Fungicida - Candida 2-Cloro-N-Fenilacetamida Antibiofilmes Docking molecular Dinâmica molecular Fungicide - Candida 2-Chloro-N-Phenylacetamide Antibiofilms Molecular docking Molecular dinamic CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
topic |
Fungicida - Candida 2-Cloro-N-Fenilacetamida Antibiofilmes Docking molecular Dinâmica molecular Fungicide - Candida 2-Chloro-N-Phenylacetamide Antibiofilms Molecular docking Molecular dinamic CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
description |
Candida and mycoses are a growing global public health concern. Candida albicans, Candida tropicalis and Candida parapsilosis are on the list of priority fungal pathogens created by the World Health Organization. Thus, the development of research is planned to generate new drugs in order to combat and prevent the development of resistance. In this sense, prospective studies with reaction intermediates with amide groups and chlorine atoms are a promising strategy. This work aimed to evaluate the antifungal activity of the synthetic molecule 2-Chloro- N-phenylacetamides (A1Cl) against C. albicans, C. tropicalis and C. parapsilosis. The minimum inhibitory concentration (MIC) was determined by microdilution, so that values ranged from 16 to 256 μg/mL for A1Cl and 8 to 512 μg/mL for fluconazole (positive control). The minimum fungicidal concentration was similar to the MIC, for both evaluated compounds, confirming the fungicidal effect of A1Cl. In the model of morphological changes (microculture technique), A1Cl and fluconazole reduced the formation of virulence structures compared to their respective controls, showing a concentration-dependent effect. The presence of sorbitol and exogenous ergosterol did not alter the MIC values, confirming that A1Cl has no effect interfering with the functionality and/or integrity of the cell wall and membrane, respectively. The association study (checkerboard) of A1Cl with fluconazole resulted in an antifungal effect of indifference. In the in vitro antibiofilm effect assay, A1Cl was able to inhibit biofilm formation by more than 45% and 80% (***p<0.001), and was able to promote preformed biofilm rupture by more than 35% and 60 % (***p<0.001), in sub and supra inhibitory concentrations, respectively, when compared to the respective control groups. However, for fluconazole to achieve 50% (***p<0.001) or more inhibition effect, suprainhibitory concentrations were required. Similarly, A1Cl (CIM) was able to inhibit by more than 50% (***p<0.001) the formation of ex vivo biofilms (nail fragments) and for fluconazole to achieve the same antibiofilm effect by more than 50% 4xCIM was required. A1Cl showed binding energy values greater than or close to fluconazole in at least one scoring function, for Dihydrofolate reductase (DHFR), Geranylgeranyltransferase-I (GGTase-I) and Aspartic Protease-2 (SAP-2), so that these enzymes, may be targets susceptible to the antifungal action of A1Cl. The RMSD of the evaluated complexes revealed stability around 2-3ns and maintained its stability above 10 ns. For the RMSF of the complexes, we observed interactions similar to those investigated in docking after dynamic simulations at times of 200 and 600ps and that A1Cl remained in the active site under the influence of solvents and structural flexibility. Given these results, it is possible to infer that A1Cl has an antifungal effect and that this activity is related to antibiofilm mechanisms and binding to DHFR and SAP-2. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-07-20 2023-03-03 2024-03-06T13:01:11Z 2024-03-06T13:01:11Z |
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/123456789/29761 |
url |
https://repositorio.ufpb.br/jspui/handle/123456789/29761 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
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 |
dc.source.none.fl_str_mv |
reponame:Biblioteca Digital de Teses e Dissertações da UFPB instname:Universidade Federal da Paraíba (UFPB) instacron:UFPB |
instname_str |
Universidade Federal da Paraíba (UFPB) |
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UFPB |
institution |
UFPB |
reponame_str |
Biblioteca Digital de Teses e Dissertações da UFPB |
collection |
Biblioteca Digital de Teses e Dissertações da UFPB |
repository.name.fl_str_mv |
Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB) |
repository.mail.fl_str_mv |
diretoria@ufpb.br|| diretoria@ufpb.br |
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1801843030018228224 |