Toxicidade e potencial antitumoral do derivado espiro-acridínico (E)-1'-((4- clorobenzilideno)amino)-5'-oxo-1',5'-diidro-10H-espiro[acridina-9,2'-pirrol]- 4'-carbonitrila (AMTAC-06)
Autor(a) principal: | |
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Data de Publicação: | 2021 |
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/123456789/20981 |
Resumo: | Cancer comprise an esemble of diseases characterized by uncontrolled cell growth and metastatic potential. Current chemotherapy treatments have limitations, mainly due to toxicity and the development of tumor resistance. Considering that acridine compounds are reported as promising anticancer agents, this study aimed to investigate the antitumor activity and toxicity of the new synthetic spiro-acridine compound (E)-1'-((4-chlorobenzylidene)amino)-5'- oxo-1',5'-dihydro-10H-spiro[acridine-9,2'-pyrrole]-4'-carbonitrile (AMTAC-06), selected after pharmacological screening. In vitro antitumor activity and cytotoxicity was evaluated by the MTT assay, using tumor cells (HCT-116, HeLa, MCF-7, PC-3, MDA-MB-231, SK-MEL-28, HL-60) and non-tumor cells (L929, HaCat, PBMC). AMTAC-06 induced higher cytotoxicity in colorectal carcinoma cells, HCT-116 (IC50, or half-maximal inhibitory concentration, of 12.62 µM, in 72 hours), and reduced the viability of non-tumor cells (HaCaTIC50: 17.87 µM; L929IC50: 26.15 µM; PBMCIC50: 7.89 µM). However, AMTAC-06 was less cytotoxic compared to the standard drug doxorubicin (HCT-116IC50: 2.57 µM; HaCaTIC50: 0.28 µM; PBMCIC50: 0.05 µM). In order to elucidate its mechanisms of action in vitro, the effects on cell cycle, apoptosis and on the production of reactive oxygen species (ROS) in HCT-116 cells (15 and 30 µM AMTAC-06, after 48 hours) were evaluated. AMTAC-06 induced an increase in the sub-G1 fraction and cell cycle arrest in the S phase (p <0.05). Morphological characteristics of apoptosis, such as membrane blebbing formation, apoptotic bodies, chromatin condensation and nuclear fragmentation, were observed by confocal microscopy. Simultaneously, there was an increase in the number of staining cells with annexin V (p <0.05), characterizing apoptosis. There was also a reduction in the production of ROS (p <0.05), which suggests an antioxidant effect. In vivo, AMTAC-06 toxicity was investigated in Swiss mice (Mus Musculus) and in zebrafish (Danio rerio), and the antitumor activity was studied using the Ehrlich Ascites Carcinoma (EAC) model. AMTAC-06 did not induce toxicity in zebrafish embryos/larvae (LC50, or mean lethal concentration, higher than 126.2 µM) or in mice (LD50, or mean lethal dose, higher than 5000 mg/kg, i.p.). Genotoxicity was assessed by the micronucleus test on peripheral blood of mices, and it was observed that AMTAC-06 (2000 mg/kg, i.p.) did not show genotoxicity. In a EAC model, AMTAC-06 (12.5 mg/kg, i.p.) reduced the viability and the total peritoneal tumor cells (p <0.05), as well as the microdensity of the peritumoral vessels (p <0.05), indicating antiangiogenic effect. In addition, there was an increase in the levels of cytokines TNF-α and IL-1β, as well as a reduction in INF- in the peritoneal fluid, characterizing an immunomodulatory action associated with antitumor activity. The analysis of biochemical, hematological and histological parameters in animals transplanted with Ehrlich and treated with AMTAC-06 (12.5 mg/kg, i.p.) did not show toxicity. In conclusion, the new spiro-acridine derivative AMTAC-06 has antitumor activity in vitro and in vivo, with low toxicity, which indicates its potential as an anticancer agent. |
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Toxicidade e potencial antitumoral do derivado espiro-acridínico (E)-1'-((4- clorobenzilideno)amino)-5'-oxo-1',5'-diidro-10H-espiro[acridina-9,2'-pirrol]- 4'-carbonitrila (AMTAC-06)Espiro-acridínicoAtividade antitumoralToxicidadeCarcinoma colorretalPeixe-zebraCarcinoma ascítico de Ehrlich.Spiro-acridineAntitumor activityToxicityColorectal carcinomaZebrafishEhrlich ascites carcinomaCNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIACancer comprise an esemble of diseases characterized by uncontrolled cell growth and metastatic potential. Current chemotherapy treatments have limitations, mainly due to toxicity and the development of tumor resistance. Considering that acridine compounds are reported as promising anticancer agents, this study aimed to investigate the antitumor activity and toxicity of the new synthetic spiro-acridine compound (E)-1'-((4-chlorobenzylidene)amino)-5'- oxo-1',5'-dihydro-10H-spiro[acridine-9,2'-pyrrole]-4'-carbonitrile (AMTAC-06), selected after pharmacological screening. In vitro antitumor activity and cytotoxicity was evaluated by the MTT assay, using tumor cells (HCT-116, HeLa, MCF-7, PC-3, MDA-MB-231, SK-MEL-28, HL-60) and non-tumor cells (L929, HaCat, PBMC). AMTAC-06 induced higher cytotoxicity in colorectal carcinoma cells, HCT-116 (IC50, or half-maximal inhibitory concentration, of 12.62 µM, in 72 hours), and reduced the viability of non-tumor cells (HaCaTIC50: 17.87 µM; L929IC50: 26.15 µM; PBMCIC50: 7.89 µM). However, AMTAC-06 was less cytotoxic compared to the standard drug doxorubicin (HCT-116IC50: 2.57 µM; HaCaTIC50: 0.28 µM; PBMCIC50: 0.05 µM). In order to elucidate its mechanisms of action in vitro, the effects on cell cycle, apoptosis and on the production of reactive oxygen species (ROS) in HCT-116 cells (15 and 30 µM AMTAC-06, after 48 hours) were evaluated. AMTAC-06 induced an increase in the sub-G1 fraction and cell cycle arrest in the S phase (p <0.05). Morphological characteristics of apoptosis, such as membrane blebbing formation, apoptotic bodies, chromatin condensation and nuclear fragmentation, were observed by confocal microscopy. Simultaneously, there was an increase in the number of staining cells with annexin V (p <0.05), characterizing apoptosis. There was also a reduction in the production of ROS (p <0.05), which suggests an antioxidant effect. In vivo, AMTAC-06 toxicity was investigated in Swiss mice (Mus Musculus) and in zebrafish (Danio rerio), and the antitumor activity was studied using the Ehrlich Ascites Carcinoma (EAC) model. AMTAC-06 did not induce toxicity in zebrafish embryos/larvae (LC50, or mean lethal concentration, higher than 126.2 µM) or in mice (LD50, or mean lethal dose, higher than 5000 mg/kg, i.p.). Genotoxicity was assessed by the micronucleus test on peripheral blood of mices, and it was observed that AMTAC-06 (2000 mg/kg, i.p.) did not show genotoxicity. In a EAC model, AMTAC-06 (12.5 mg/kg, i.p.) reduced the viability and the total peritoneal tumor cells (p <0.05), as well as the microdensity of the peritumoral vessels (p <0.05), indicating antiangiogenic effect. In addition, there was an increase in the levels of cytokines TNF-α and IL-1β, as well as a reduction in INF- in the peritoneal fluid, characterizing an immunomodulatory action associated with antitumor activity. The analysis of biochemical, hematological and histological parameters in animals transplanted with Ehrlich and treated with AMTAC-06 (12.5 mg/kg, i.p.) did not show toxicity. In conclusion, the new spiro-acridine derivative AMTAC-06 has antitumor activity in vitro and in vivo, with low toxicity, which indicates its potential as an anticancer agent.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESO câncer compreende um conjunto de doenças caracterizadas pelo crescimento celular descontrolado e potencial metastático. Os tratamentos quimioterápicos atuais apresentam limitações, principalmente devido à toxicidade e ao desenvolvimento de resistência tumoral. Considerando que os compostos acridínicos são relatados como promissores agentes anticâncer, este trabalho teve como objetivo investigar a atividade antitumoral e a toxicidade do novo composto espiro-acridínico sintético (E)-1'-((4- clorobenzilideno)amino)-5'-oxo-1',5'-diidro-10H-espiro[acridina-9,2'-pirrol]-4'- carbonitrila (AMTAC-06), selecionado após triagem farmacológica. A atividade antitumoral in vitro e citotoxicidade foram avaliados pelo ensaio do MTT, utilizando células tumorais (HCT-116, HeLa, MCF-7, PC-3, MDA-MB-231, SKMEL-28, HL-60) e não tumorais (L929, HaCat, PBMC). AMTAC-06 induziu maior citotoxicidade em células de carcinoma colorretal, HCT-116 (CI50, ou concentração inibitória média, de 12,62 µM, em 72 horas), e reduziu a viabilidade das células não tumorais (HaCaTCI50: 17,87 µM; L929CI50: 26,15 µM; PBMCCI50: 7,89 µM). Todavia, o AMTAC-06 foi menos citotóxico em comparação à droga padrão doxorrubicina (HCT-116CI50: 2,57 µM; HaCaTCI50: 0,28 µM; PBMCCI50: 0,05 µM). Para elucidar seus mecanismos de ação in vitro, foram avaliados os efeitos no ciclo celular, apoptose e na produção de espécies reativas de oxigênio (EROs), em células HCT-116 (15 e 30 µM de AMTAC-06, após 48 horas). AMTAC-06 induziu aumento na fração sub-G1 e parada do ciclo celular na fase S (p<0,05). Características morfológicas de apoptose, como formação de blebs na membrana, corpos apoptóticos, condensação da cromatina e fragmentação nuclear, foram observadas por microscopia confocal. Em paralelo, ocorreu aumento na quantidade de células marcadas com anexina V (p<0,05), caracterizando apoptose. Ainda, houve redução na produção de EROs (p<0,05), o que sugere efeito antioxidante. In vivo, a toxicidade do AMTAC-06 foi investigada em camundongos Swiss (Mus Musculus) e em peixe-zebra (Danio rerio), e a atividade antitumoral foi estudada usando o modelo de Carcinoma Ascítico de Ehrlich (CAE). AMTAC06 não induziu toxicidade em embriões/larvas de peixe-zebra (CL50, ou concentração letal média, superior a 126,2 µM) e em camundongos (DL50, ou dose letal média, maior que 5000 mg/kg, i.p.). A genotoxicidade foi avaliada pelo teste do micronúcleo em sangue periférico de camundongos, sendo observado que AMTAC-06 (2000 mg/kg, i.p.) não apresentou genotoxicidade. Em modelo de CAE, AMTAC-06 (12,5 mg/kg, i.p.) reduziu a viabilidade e o total de células tumorais peritoneais (p<0,05), bem como a microdensidade dos vasos peritumorais (p<0,05), indicando efeito antiangiogênico. Ainda, houve aumento nos níveis das citocinas TNF-α e IL-1β, bem como redução de INF- no fluido peritoneal, caracterizando uma ação imunomoduladora associada à atividade antitumoral. A análise de parâmetros bioquímicos, hematológicos e histológicos nos animais transplantados com Ehrlich e tratados com AMTAC-06 (12,5 mg/kg, i.p.) não evidenciou toxicidade. Em conclusão, o novo derivado espiro-acridínico AMTAC-06 apresenta atividade antitumoral in vitro e in vivo, com baixa toxicidade, o que indica seu potencial como um agente anticâncer.Universidade Federal da ParaíbaBrasilCiências BiológicasPrograma de Pós-Graduação em Produtos Naturais e Sintéticos BioativosUFPBSobral, Marianna Vieirahttp://lattes.cnpq.br/1036684849301560Duarte, Sâmia Sousa2021-09-14T19:27:11Z2021-03-172021-09-14T19:27:11Z2021-02-05info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesishttps://repositorio.ufpb.br/jspui/handle/123456789/20981porAttribution-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:UFPB2022-08-09T18:23:56Zoai:repositorio.ufpb.br:123456789/20981Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufpb.br/PUBhttp://tede.biblioteca.ufpb.br:8080/oai/requestdiretoria@ufpb.br|| diretoria@ufpb.bropendoar:2022-08-09T18:23:56Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB)false |
dc.title.none.fl_str_mv |
Toxicidade e potencial antitumoral do derivado espiro-acridínico (E)-1'-((4- clorobenzilideno)amino)-5'-oxo-1',5'-diidro-10H-espiro[acridina-9,2'-pirrol]- 4'-carbonitrila (AMTAC-06) |
title |
Toxicidade e potencial antitumoral do derivado espiro-acridínico (E)-1'-((4- clorobenzilideno)amino)-5'-oxo-1',5'-diidro-10H-espiro[acridina-9,2'-pirrol]- 4'-carbonitrila (AMTAC-06) |
spellingShingle |
Toxicidade e potencial antitumoral do derivado espiro-acridínico (E)-1'-((4- clorobenzilideno)amino)-5'-oxo-1',5'-diidro-10H-espiro[acridina-9,2'-pirrol]- 4'-carbonitrila (AMTAC-06) Duarte, Sâmia Sousa Espiro-acridínico Atividade antitumoral Toxicidade Carcinoma colorretal Peixe-zebra Carcinoma ascítico de Ehrlich. Spiro-acridine Antitumor activity Toxicity Colorectal carcinoma Zebrafish Ehrlich ascites carcinoma CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
title_short |
Toxicidade e potencial antitumoral do derivado espiro-acridínico (E)-1'-((4- clorobenzilideno)amino)-5'-oxo-1',5'-diidro-10H-espiro[acridina-9,2'-pirrol]- 4'-carbonitrila (AMTAC-06) |
title_full |
Toxicidade e potencial antitumoral do derivado espiro-acridínico (E)-1'-((4- clorobenzilideno)amino)-5'-oxo-1',5'-diidro-10H-espiro[acridina-9,2'-pirrol]- 4'-carbonitrila (AMTAC-06) |
title_fullStr |
Toxicidade e potencial antitumoral do derivado espiro-acridínico (E)-1'-((4- clorobenzilideno)amino)-5'-oxo-1',5'-diidro-10H-espiro[acridina-9,2'-pirrol]- 4'-carbonitrila (AMTAC-06) |
title_full_unstemmed |
Toxicidade e potencial antitumoral do derivado espiro-acridínico (E)-1'-((4- clorobenzilideno)amino)-5'-oxo-1',5'-diidro-10H-espiro[acridina-9,2'-pirrol]- 4'-carbonitrila (AMTAC-06) |
title_sort |
Toxicidade e potencial antitumoral do derivado espiro-acridínico (E)-1'-((4- clorobenzilideno)amino)-5'-oxo-1',5'-diidro-10H-espiro[acridina-9,2'-pirrol]- 4'-carbonitrila (AMTAC-06) |
author |
Duarte, Sâmia Sousa |
author_facet |
Duarte, Sâmia Sousa |
author_role |
author |
dc.contributor.none.fl_str_mv |
Sobral, Marianna Vieira http://lattes.cnpq.br/1036684849301560 |
dc.contributor.author.fl_str_mv |
Duarte, Sâmia Sousa |
dc.subject.por.fl_str_mv |
Espiro-acridínico Atividade antitumoral Toxicidade Carcinoma colorretal Peixe-zebra Carcinoma ascítico de Ehrlich. Spiro-acridine Antitumor activity Toxicity Colorectal carcinoma Zebrafish Ehrlich ascites carcinoma CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
topic |
Espiro-acridínico Atividade antitumoral Toxicidade Carcinoma colorretal Peixe-zebra Carcinoma ascítico de Ehrlich. Spiro-acridine Antitumor activity Toxicity Colorectal carcinoma Zebrafish Ehrlich ascites carcinoma CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
description |
Cancer comprise an esemble of diseases characterized by uncontrolled cell growth and metastatic potential. Current chemotherapy treatments have limitations, mainly due to toxicity and the development of tumor resistance. Considering that acridine compounds are reported as promising anticancer agents, this study aimed to investigate the antitumor activity and toxicity of the new synthetic spiro-acridine compound (E)-1'-((4-chlorobenzylidene)amino)-5'- oxo-1',5'-dihydro-10H-spiro[acridine-9,2'-pyrrole]-4'-carbonitrile (AMTAC-06), selected after pharmacological screening. In vitro antitumor activity and cytotoxicity was evaluated by the MTT assay, using tumor cells (HCT-116, HeLa, MCF-7, PC-3, MDA-MB-231, SK-MEL-28, HL-60) and non-tumor cells (L929, HaCat, PBMC). AMTAC-06 induced higher cytotoxicity in colorectal carcinoma cells, HCT-116 (IC50, or half-maximal inhibitory concentration, of 12.62 µM, in 72 hours), and reduced the viability of non-tumor cells (HaCaTIC50: 17.87 µM; L929IC50: 26.15 µM; PBMCIC50: 7.89 µM). However, AMTAC-06 was less cytotoxic compared to the standard drug doxorubicin (HCT-116IC50: 2.57 µM; HaCaTIC50: 0.28 µM; PBMCIC50: 0.05 µM). In order to elucidate its mechanisms of action in vitro, the effects on cell cycle, apoptosis and on the production of reactive oxygen species (ROS) in HCT-116 cells (15 and 30 µM AMTAC-06, after 48 hours) were evaluated. AMTAC-06 induced an increase in the sub-G1 fraction and cell cycle arrest in the S phase (p <0.05). Morphological characteristics of apoptosis, such as membrane blebbing formation, apoptotic bodies, chromatin condensation and nuclear fragmentation, were observed by confocal microscopy. Simultaneously, there was an increase in the number of staining cells with annexin V (p <0.05), characterizing apoptosis. There was also a reduction in the production of ROS (p <0.05), which suggests an antioxidant effect. In vivo, AMTAC-06 toxicity was investigated in Swiss mice (Mus Musculus) and in zebrafish (Danio rerio), and the antitumor activity was studied using the Ehrlich Ascites Carcinoma (EAC) model. AMTAC-06 did not induce toxicity in zebrafish embryos/larvae (LC50, or mean lethal concentration, higher than 126.2 µM) or in mice (LD50, or mean lethal dose, higher than 5000 mg/kg, i.p.). Genotoxicity was assessed by the micronucleus test on peripheral blood of mices, and it was observed that AMTAC-06 (2000 mg/kg, i.p.) did not show genotoxicity. In a EAC model, AMTAC-06 (12.5 mg/kg, i.p.) reduced the viability and the total peritoneal tumor cells (p <0.05), as well as the microdensity of the peritumoral vessels (p <0.05), indicating antiangiogenic effect. In addition, there was an increase in the levels of cytokines TNF-α and IL-1β, as well as a reduction in INF- in the peritoneal fluid, characterizing an immunomodulatory action associated with antitumor activity. The analysis of biochemical, hematological and histological parameters in animals transplanted with Ehrlich and treated with AMTAC-06 (12.5 mg/kg, i.p.) did not show toxicity. In conclusion, the new spiro-acridine derivative AMTAC-06 has antitumor activity in vitro and in vivo, with low toxicity, which indicates its potential as an anticancer agent. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-09-14T19:27:11Z 2021-03-17 2021-09-14T19:27:11Z 2021-02-05 |
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/123456789/20981 |
url |
https://repositorio.ufpb.br/jspui/handle/123456789/20981 |
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 Ciências Biológicas Programa de Pós-Graduação em Produtos Naturais e Sintéticos Bioativos UFPB |
publisher.none.fl_str_mv |
Universidade Federal da Paraíba Brasil Ciências Biológicas 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) |
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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1801842980368154624 |