Investigação da atividade biológicas dos adutos de Morita-Baylis-Hillman sobre Leishmania spp
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da UFPB |
Texto Completo: | https://repositorio.ufpb.br/jspui/handle/123456789/13654 |
Resumo: | Leishmaniasis treatment is based on the use of pentavalent antimonial compounds, as first choice drugs; even though they are highly toxic presenting several side effects. This diseases complex is a public health problem, with challenges in the new therapies identification that allow a greater adhesion and better quality in the treatment of the patients. Due to high toxicity and drug resistance, the search of new alternatives for the treatment of leishmaniasis is justified. In this context, this study aimed to evaluate the anti-Leishmania activity of Morita-Baylis-Hillman adducts in vitro and ex vivo experimental models. Activity of the compounds on the promastigote and axenic amastigote forms of the Leishmania parasite, in addition to the human peripheral blood monuclear cells, were evaluated by the MTT colorimetric test (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium). The rate of infection in human monocytes and the cell death profile of promastigote forms were assessed by flow cytometry. In addition, hemolytic activity on human erythrocytes and indirect evaluation of nitric oxide levels by Griess's reagent were measured. Screening of 36 molecules from the Morita-Baylis-Hillman reaction on L. donovani's promastigote forms had MBH-A12 and MBH-A13 as one of the lowest IC50 values (4.71 and 0.37 μg/mL), besides they present the same chemical synthesis process, being thus selected for the continuation of the biological evaluation. These two substances also showed significant inhibition of growth (IC50) on the promastigote forms of L. infantum (5.6 and 5.3 μg/mL), L. amazonensis (13.2 and 13.4 μg/mL) and L. braziliensis (16.32 and 19.09 μg/mL). In addition, they showed activity on the axenic amastigote forms of L. infantum (EC50 of 9.08 and 10.76 μg/mL) and L. amazonensis (EC50 of 13.4 and 10.4 μg/mL), demonstrating effect on the diversity of Leishmania species that cause different clinical manifestations. Among the tested concentrations, these drugs did not present toxicity to peripheral blood mononuclear cells and human erythrocytes, thus demonstrating selectivity indices to parasite tropism. In the infection model evaluation of human monocytes, we did not observe statistical difference with the substances treatment, nor in modulation via nitric oxide synthesis. Cell death profile in Leishmania spp. suggests a pattern involving apoptosis death mechanism for MBH-A12 and MBH-A13 adducts, different from amphotericin B which also leads to the necrosis process. Thus, we concluded that the Morita-BaylisHillman adducts are compounds with expressive anti-Leishmania activity in an in vitro model, being molecules that need to be better investigated, regarding to the action mechanism, in the search for new treatments for leishmaniasis. |
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Investigação da atividade biológicas dos adutos de Morita-Baylis-Hillman sobre Leishmania spp Atividade anti-LeishmaniaCompostos sintéticosLeishmaniosesAnti-Leishmania activitySynthetic compoundsLeishmaniasisLeishmaniose - TratamentoAtividade anti-leishmaniaMorita-Baylis-HillmanCNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIALeishmaniasis treatment is based on the use of pentavalent antimonial compounds, as first choice drugs; even though they are highly toxic presenting several side effects. This diseases complex is a public health problem, with challenges in the new therapies identification that allow a greater adhesion and better quality in the treatment of the patients. Due to high toxicity and drug resistance, the search of new alternatives for the treatment of leishmaniasis is justified. In this context, this study aimed to evaluate the anti-Leishmania activity of Morita-Baylis-Hillman adducts in vitro and ex vivo experimental models. Activity of the compounds on the promastigote and axenic amastigote forms of the Leishmania parasite, in addition to the human peripheral blood monuclear cells, were evaluated by the MTT colorimetric test (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium). The rate of infection in human monocytes and the cell death profile of promastigote forms were assessed by flow cytometry. In addition, hemolytic activity on human erythrocytes and indirect evaluation of nitric oxide levels by Griess's reagent were measured. Screening of 36 molecules from the Morita-Baylis-Hillman reaction on L. donovani's promastigote forms had MBH-A12 and MBH-A13 as one of the lowest IC50 values (4.71 and 0.37 μg/mL), besides they present the same chemical synthesis process, being thus selected for the continuation of the biological evaluation. These two substances also showed significant inhibition of growth (IC50) on the promastigote forms of L. infantum (5.6 and 5.3 μg/mL), L. amazonensis (13.2 and 13.4 μg/mL) and L. braziliensis (16.32 and 19.09 μg/mL). In addition, they showed activity on the axenic amastigote forms of L. infantum (EC50 of 9.08 and 10.76 μg/mL) and L. amazonensis (EC50 of 13.4 and 10.4 μg/mL), demonstrating effect on the diversity of Leishmania species that cause different clinical manifestations. Among the tested concentrations, these drugs did not present toxicity to peripheral blood mononuclear cells and human erythrocytes, thus demonstrating selectivity indices to parasite tropism. In the infection model evaluation of human monocytes, we did not observe statistical difference with the substances treatment, nor in modulation via nitric oxide synthesis. Cell death profile in Leishmania spp. suggests a pattern involving apoptosis death mechanism for MBH-A12 and MBH-A13 adducts, different from amphotericin B which also leads to the necrosis process. Thus, we concluded that the Morita-BaylisHillman adducts are compounds with expressive anti-Leishmania activity in an in vitro model, being molecules that need to be better investigated, regarding to the action mechanism, in the search for new treatments for leishmaniasis.O tratamento das leishmanioses é baseado na utilização de compostos antimoniais pentavalentes, como medicamentos de primeira escolha; sendo drogas altamente tóxicas e que apresentam vários efeitos colaterais. Esse complexo de doenças é um problema de saúde pública, com desafios na identificação de novas terapias que permitam uma maior adesão e melhor qualidade no tratamento aos pacientes. Com a elevada toxicidade e o aparecimento de resistência às drogas, se justifica a busca de novas alternativas para o tratamento das leishmanioses. Neste contexto, este trabalho teve como objetivo avaliar a atividade anti-Leishmania dos adutos de Morita-Baylis-Hillman em modelos experimentais in vitro e ex vivo. A atividade dos compostos sobre as formas promastigotas e amastigotas axênicas do parasito Leishmania, além das células monucleares de sangue periférico humano foram avaliadas pelo teste colorimétrico do MTT (brometo de 3-(4,5-dimetiltiazol-2-yl)-2,5difenil tetrazólio). A taxa de infecção em monócitos humanos e o perfil de morte celular de formas promastigotas foram avaliados por citometria de fluxo. Além disso, foi mensurada a atividade hemolítica sobre eritrócitos humanos e a avaliação indireta dos níveis de óxido nítrico pelo reagente de Griess. A triagem das 36 moléculas oriundas da reação de Morita-Baylis-Hillman sobre as formas promastigotas de L. donovani, teve os MBH-A12 e MBH-A13 com um dos menores valores de CI50 (4,71 e 0,37 µg/mL), além de que apresentam o mesmo processo de síntese química, sendo assim selecionados para a continuação da avaliação biológica. Essas duas substâncias também apresentaram inibição de crescimento (CI50) significativa sobre as formas promastigotas de L. infantum (5,6 e 5,3 µg/mL), L. amazonensis (13,2 e 13,4 µg/mL) e L. braziliensis (16,32 e 19,09 µg/mL); além disso, apresentaram atividade sobre as formas amastigotas axênicas de L. infantum (CE50 de 9,08 e 10,76 µg/mL) e L. amazonensis (CE50 de 13,4 e 10,4 µg/mL), demonstrando efeito sobre a diversidade de espécies de Leishmania que causam manifestações clínicas diferentes. Dentro das concentrações testadas, essas drogas não apresentaram toxicidade para células mononucleares de sangue periférico e eritrócitos humanos, apresentando assim índices de seletividade com tropismo para o parasito. No modelo de avaliação da taxa de infecção em monócitos humanos, não observamos diferença estatística com o tratamento das substâncias, nem modulação via síntese de óxido nítrico. O perfil de morte celular em promastigotas de Leishmania spp. sugere um padrão envolvendo mecanismo de morte via apoptose para os adutos MBH-A12 e MBH-A13, diferente da anfotericina B que também leva ao processo de necrose. Assim, conclui-se que os adutos de MoritaBaylis-Hillman são compostos com atividade anti-Leishmania em modelo in vitro, sendo moléculas a serem investigadas quanto ao mecanismo de ação, na busca por novos tratamentos para as leishmanioses.Universidade Federal da ParaíbaBrasilFarmacologiaPrograma de Pós-Graduação em Produtos Naturais e Sintéticos BioativosUFPBCruz , Josiane de Camposhttp://lattes.cnpq.br/7245505923489654Clemente , Tatjana Keesen de Souza Limahttp://lattes.cnpq.br/5504382837656473Rocha, Juliana da Câmara2019-02-21T15:05:36Z2018-05-102019-02-21T15:05:36Z2018-05-04info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesishttps://repositorio.ufpb.br/jspui/handle/123456789/13654porAttribution-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:UFPB2019-02-22T06:03:10Zoai:repositorio.ufpb.br:123456789/13654Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufpb.br/PUBhttp://tede.biblioteca.ufpb.br:8080/oai/requestdiretoria@ufpb.br|| diretoria@ufpb.bropendoar:2019-02-22T06:03:10Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB)false |
dc.title.none.fl_str_mv |
Investigação da atividade biológicas dos adutos de Morita-Baylis-Hillman sobre Leishmania spp |
title |
Investigação da atividade biológicas dos adutos de Morita-Baylis-Hillman sobre Leishmania spp |
spellingShingle |
Investigação da atividade biológicas dos adutos de Morita-Baylis-Hillman sobre Leishmania spp Rocha, Juliana da Câmara Atividade anti-Leishmania Compostos sintéticos Leishmanioses Anti-Leishmania activity Synthetic compounds Leishmaniasis Leishmaniose - Tratamento Atividade anti-leishmania Morita-Baylis-Hillman CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
title_short |
Investigação da atividade biológicas dos adutos de Morita-Baylis-Hillman sobre Leishmania spp |
title_full |
Investigação da atividade biológicas dos adutos de Morita-Baylis-Hillman sobre Leishmania spp |
title_fullStr |
Investigação da atividade biológicas dos adutos de Morita-Baylis-Hillman sobre Leishmania spp |
title_full_unstemmed |
Investigação da atividade biológicas dos adutos de Morita-Baylis-Hillman sobre Leishmania spp |
title_sort |
Investigação da atividade biológicas dos adutos de Morita-Baylis-Hillman sobre Leishmania spp |
author |
Rocha, Juliana da Câmara |
author_facet |
Rocha, Juliana da Câmara |
author_role |
author |
dc.contributor.none.fl_str_mv |
Cruz , Josiane de Campos http://lattes.cnpq.br/7245505923489654 Clemente , Tatjana Keesen de Souza Lima http://lattes.cnpq.br/5504382837656473 |
dc.contributor.author.fl_str_mv |
Rocha, Juliana da Câmara |
dc.subject.por.fl_str_mv |
Atividade anti-Leishmania Compostos sintéticos Leishmanioses Anti-Leishmania activity Synthetic compounds Leishmaniasis Leishmaniose - Tratamento Atividade anti-leishmania Morita-Baylis-Hillman CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
topic |
Atividade anti-Leishmania Compostos sintéticos Leishmanioses Anti-Leishmania activity Synthetic compounds Leishmaniasis Leishmaniose - Tratamento Atividade anti-leishmania Morita-Baylis-Hillman CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
description |
Leishmaniasis treatment is based on the use of pentavalent antimonial compounds, as first choice drugs; even though they are highly toxic presenting several side effects. This diseases complex is a public health problem, with challenges in the new therapies identification that allow a greater adhesion and better quality in the treatment of the patients. Due to high toxicity and drug resistance, the search of new alternatives for the treatment of leishmaniasis is justified. In this context, this study aimed to evaluate the anti-Leishmania activity of Morita-Baylis-Hillman adducts in vitro and ex vivo experimental models. Activity of the compounds on the promastigote and axenic amastigote forms of the Leishmania parasite, in addition to the human peripheral blood monuclear cells, were evaluated by the MTT colorimetric test (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium). The rate of infection in human monocytes and the cell death profile of promastigote forms were assessed by flow cytometry. In addition, hemolytic activity on human erythrocytes and indirect evaluation of nitric oxide levels by Griess's reagent were measured. Screening of 36 molecules from the Morita-Baylis-Hillman reaction on L. donovani's promastigote forms had MBH-A12 and MBH-A13 as one of the lowest IC50 values (4.71 and 0.37 μg/mL), besides they present the same chemical synthesis process, being thus selected for the continuation of the biological evaluation. These two substances also showed significant inhibition of growth (IC50) on the promastigote forms of L. infantum (5.6 and 5.3 μg/mL), L. amazonensis (13.2 and 13.4 μg/mL) and L. braziliensis (16.32 and 19.09 μg/mL). In addition, they showed activity on the axenic amastigote forms of L. infantum (EC50 of 9.08 and 10.76 μg/mL) and L. amazonensis (EC50 of 13.4 and 10.4 μg/mL), demonstrating effect on the diversity of Leishmania species that cause different clinical manifestations. Among the tested concentrations, these drugs did not present toxicity to peripheral blood mononuclear cells and human erythrocytes, thus demonstrating selectivity indices to parasite tropism. In the infection model evaluation of human monocytes, we did not observe statistical difference with the substances treatment, nor in modulation via nitric oxide synthesis. Cell death profile in Leishmania spp. suggests a pattern involving apoptosis death mechanism for MBH-A12 and MBH-A13 adducts, different from amphotericin B which also leads to the necrosis process. Thus, we concluded that the Morita-BaylisHillman adducts are compounds with expressive anti-Leishmania activity in an in vitro model, being molecules that need to be better investigated, regarding to the action mechanism, in the search for new treatments for leishmaniasis. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-05-10 2018-05-04 2019-02-21T15:05:36Z 2019-02-21T15:05:36Z |
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.uri.fl_str_mv |
https://repositorio.ufpb.br/jspui/handle/123456789/13654 |
url |
https://repositorio.ufpb.br/jspui/handle/123456789/13654 |
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 |
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Universidade Federal da Paraíba (UFPB) |
instacron_str |
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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1801842945248198656 |