Contribuição ao conhecimento fitoquímico de Cissampelos sympodialis Eichl. (Menispermaceae)

Detalhes bibliográficos
Autor(a) principal: Lima, Natanael Teles Ramos de
Data de Publicação: 2020
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/123456789/18499
Resumo: Cissampelos sympodialis Eichl., popularly known as “milona”, “jarrinha”, “abuteira” and “orelha-de-onça”, it is a species rich in alkaloids and has traditional use in the treatment of asthma, bronchitis, rheumatism and arthritis. Thus, the objective of this work was to contribute to the chemical and pharmacological knowledge of the species, through the isolation of substances, mass spectrometry, in silico studies and biological tests. The aerial parts were collected in the garden of UFPB’s Research Institute for Drugs and Medicines – IpeFarM and an exsiccate is deposited in the Herbarium Prof. Lauro Pires Xavier under AGRA 1476 registration and in the National System of Management of Genetic Heritage and Associated Traditional Knowledge (SisGen) under the code A9B2EFC. After harvest, drying and grinding, extraction was performed with 95% ethanol and then the solvent was concentrated in a rotary evaporator, obtaining the Crude Ethanol Extract (CEE). The CEE was partitioned using hexane, chloroform and ethyl acetate. The hydromethanolic phase was subjected to open column chromatography using Sephadex-LH 20, resulting in the CsFHM 6-7 fraction that was processed by preparative High Performance Liquid Chromatography (HPLC). An alkaloids march was performed to obtain the Alkaloid Rich Fraction (ARF). The isolated compounds were identified using Mass spectrometry (MS), Nuclear Magnetic Resonance (NMR) and comparison with the literature. A molecular docking study was carried out on alkaloids already isolated from milona: laurifolin, milonine, warifteina, methylwarifteina, roraimina, des-7'-O-methylroraimina and epi-des-7'-Omethylroraimina; to evaluate their interactions with Topoisomerase IIα and DNA. An MTT assay was performed with samples of C. sympodialis in breast cancer cells, human melanoma, colon carcinoma, cervical adenocarcinoma and in normal human keratinocyte cells. The preparative HPLC allowed the isolation of 4 substances: the nucleoside adenosine and three flavonoids, vicenin-2, quercetin-3-O-βglycopyranosyl-(1→6)-β-galactopyranoside and canferol-3-O-β-glycopyranosyl- (1→6)-β-galactopyranoside. In addition, from the literature, 10 more compounds were identified by LC-MS: caffeic acid-hexoside, salsolinol, guanosine, syringic acid-Ohexoside, magnoflorin, benzyl alcohol-hexose-pentose, aromadendrine-hexoside, apigenin-6-C-glycoside, isoquercetin and astragalin. The docking pointed out that among milona alkaloids, roraimine is the one that best interacts with Toposiomerase IIα and with DNA and des-7'-O-methylroraimine is the best as a DNA ligand. The MTT assay indicated that CEE has inhibitory activity in colon and breast cancer cells and ARF was able to strongly inhibit the growth of all tested strains, although it has an action against normal cells. The test also demonstrated that the glycosylated flavonoids have no activity against the tested cell lines. It is concluded that milona has potential antitumor activity and that it can eventually be used as a complement in the treatment of cancer, in addition to theoretically having chemical compounds that can become active drugs against neoplasms.
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spelling Contribuição ao conhecimento fitoquímico de Cissampelos sympodialis Eichl. (Menispermaceae)MilonaCissampelosFlavonoidesDockingTopoisomeraseCitotoxicidadeFlavonoidsTopoisomeraseCytotoxicityCNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIACissampelos sympodialis Eichl., popularly known as “milona”, “jarrinha”, “abuteira” and “orelha-de-onça”, it is a species rich in alkaloids and has traditional use in the treatment of asthma, bronchitis, rheumatism and arthritis. Thus, the objective of this work was to contribute to the chemical and pharmacological knowledge of the species, through the isolation of substances, mass spectrometry, in silico studies and biological tests. The aerial parts were collected in the garden of UFPB’s Research Institute for Drugs and Medicines – IpeFarM and an exsiccate is deposited in the Herbarium Prof. Lauro Pires Xavier under AGRA 1476 registration and in the National System of Management of Genetic Heritage and Associated Traditional Knowledge (SisGen) under the code A9B2EFC. After harvest, drying and grinding, extraction was performed with 95% ethanol and then the solvent was concentrated in a rotary evaporator, obtaining the Crude Ethanol Extract (CEE). The CEE was partitioned using hexane, chloroform and ethyl acetate. The hydromethanolic phase was subjected to open column chromatography using Sephadex-LH 20, resulting in the CsFHM 6-7 fraction that was processed by preparative High Performance Liquid Chromatography (HPLC). An alkaloids march was performed to obtain the Alkaloid Rich Fraction (ARF). The isolated compounds were identified using Mass spectrometry (MS), Nuclear Magnetic Resonance (NMR) and comparison with the literature. A molecular docking study was carried out on alkaloids already isolated from milona: laurifolin, milonine, warifteina, methylwarifteina, roraimina, des-7'-O-methylroraimina and epi-des-7'-Omethylroraimina; to evaluate their interactions with Topoisomerase IIα and DNA. An MTT assay was performed with samples of C. sympodialis in breast cancer cells, human melanoma, colon carcinoma, cervical adenocarcinoma and in normal human keratinocyte cells. The preparative HPLC allowed the isolation of 4 substances: the nucleoside adenosine and three flavonoids, vicenin-2, quercetin-3-O-βglycopyranosyl-(1→6)-β-galactopyranoside and canferol-3-O-β-glycopyranosyl- (1→6)-β-galactopyranoside. In addition, from the literature, 10 more compounds were identified by LC-MS: caffeic acid-hexoside, salsolinol, guanosine, syringic acid-Ohexoside, magnoflorin, benzyl alcohol-hexose-pentose, aromadendrine-hexoside, apigenin-6-C-glycoside, isoquercetin and astragalin. The docking pointed out that among milona alkaloids, roraimine is the one that best interacts with Toposiomerase IIα and with DNA and des-7'-O-methylroraimine is the best as a DNA ligand. The MTT assay indicated that CEE has inhibitory activity in colon and breast cancer cells and ARF was able to strongly inhibit the growth of all tested strains, although it has an action against normal cells. The test also demonstrated that the glycosylated flavonoids have no activity against the tested cell lines. It is concluded that milona has potential antitumor activity and that it can eventually be used as a complement in the treatment of cancer, in addition to theoretically having chemical compounds that can become active drugs against neoplasms.Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqCissampelos sympodialis Eichl., conhecida popularmente como “milona”, “jarrinha”, “abuteira” e “orelha-de-onça”, é rica em alcaloides e tem uso tradicional no tratamento de asma, bronquite, reumatismo e artrite. Dessa forma, o objetivo deste trabalho foi contribuir com o conhecimento químico e farmacológico da espécie, por meio de isolamento de substâncias, espectrometria de massas, estudos in silico e ensaios biológicos. As partes aéreas foram coletadas no horto do Instituto de Pesquisa em Fármacos e Medicamentos – IpeFarM da UFPB e uma exsicata encontra-se depositada no Herbário Prof. Lauro Pires Xavier sob registro AGRA 1476 e no Sistema Nacional de Gestão de Patrimônio Genético e Conhecimento Tradicional Associado (SisGen) sob código A9B2EFC. Após a coleta, secagem e trituração foi feita a extração com etanol 95% e em seguida o solvente foi concentrado em rotevaporador, sendo obtido o Extrato Etanólico Bruto (EEB). O EEB foi particionado usando hexano, clorofórmio e acetato de etila. A fase hidrometanólica foi submetida à cromatografia em coluna aberta usando Sephadex-LH 20, resultando na fração CsFHM 6-7 que foi processada por Cromatografia Líquida de Alta Eficiência (CLAE) preparativa. Foi realizada uma marcha para alcaloides a fim de obter a Fração de Alcaloides Totais (FAT). Os compostos isolados foram identificados através de Espectrometria de massas (EM), Ressonância Magnética Nuclear (RMN) e comparação com a literatura. Foi realizado um estudo de docking molecular de alcaloides já isolados da milona: laurifolina, milonina, warifteina, metilwarifteina, roraimina, des-7’-O-metilroraimina e epi-des-7’-O-metilroraimina; para avaliar suas interações com a Topoisomerase IIα e com o DNA. Realizou-se um ensaio de MTT com amostras de C. sympodialis em células câncer de mama, melanoma humano, carcinoma de cólon, adenocarcinoma cervical e em células normais de queratinócitos humanos. A CLAE preparativa permitiu o isolamento de 4 substâncias: o nucleosídeo adenosina e três flavonoides, a vicenina-2, a quercetina-3-O-β-glicopiranosil-(1→6)-β-galactopiranosídeo e o canferol-3-O-β-glicopiranosil-(1→6)-β-galactopiranosídeo. Além disso, com auxílio de dados da literatura, foram identificados mais 10 compostos por CLAE-EM: ácido cafeico-hexosídeo, salsolinol, guanosina, ácido siríngico-O-hexosideo, magnoflorina, álcool benzílico-hexose-pentose, aromadendrina-hexosideo, apigenina-6-Cglicosídeo, isoquercetina e astragalina. O docking apontou que dentre os alcaloides da milona, a roraimina é a que melhor interage com a Toposiomerase IIα e com o DNA e a des-7’-O-metilroraimina é melhor como ligante de DNA. O ensaio de MTT indicou que o EEB possui atividade inibitória de células de câncer de cólon e de mama e a FAT foi capaz de inibir fortemente o crescimento de todas linhagens testadas, embora tenha apresentação ação contra células normais. O teste também demonstrou que os flavonoides glicosilados não possuem atividade contra as linhagens testadas. Concluise que a milona tem potencial atividade antitumoral e que eventualmente pode ser usada como complemento no tratamento do câncer, além de teoricamente possuir compostos químicos que podem se tornar fármacos atuantes contra neoplasias.Universidade Federal da ParaíbaBrasilFarmacologiaPrograma de Pós-Graduação em Produtos Naturais e Sintéticos BioativosUFPBBarbosa Filho, José Mariahttp://lattes.cnpq.br/8892459126928726Fechine, Ivana Mariahttp://lattes.cnpq.br/5689715833582093Lima, Natanael Teles Ramos de2020-11-23T16:55:47Z2020-07-282020-11-23T16:55:47Z2020-02-14info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesishttps://repositorio.ufpb.br/jspui/handle/123456789/18499porAttribution-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:UFPB2020-11-24T06:09:53Zoai:repositorio.ufpb.br:123456789/18499Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufpb.br/PUBhttp://tede.biblioteca.ufpb.br:8080/oai/requestdiretoria@ufpb.br|| diretoria@ufpb.bropendoar:2020-11-24T06:09:53Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB)false
dc.title.none.fl_str_mv Contribuição ao conhecimento fitoquímico de Cissampelos sympodialis Eichl. (Menispermaceae)
title Contribuição ao conhecimento fitoquímico de Cissampelos sympodialis Eichl. (Menispermaceae)
spellingShingle Contribuição ao conhecimento fitoquímico de Cissampelos sympodialis Eichl. (Menispermaceae)
Lima, Natanael Teles Ramos de
Milona
Cissampelos
Flavonoides
Docking
Topoisomerase
Citotoxicidade
Flavonoids
Topoisomerase
Cytotoxicity
CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA
title_short Contribuição ao conhecimento fitoquímico de Cissampelos sympodialis Eichl. (Menispermaceae)
title_full Contribuição ao conhecimento fitoquímico de Cissampelos sympodialis Eichl. (Menispermaceae)
title_fullStr Contribuição ao conhecimento fitoquímico de Cissampelos sympodialis Eichl. (Menispermaceae)
title_full_unstemmed Contribuição ao conhecimento fitoquímico de Cissampelos sympodialis Eichl. (Menispermaceae)
title_sort Contribuição ao conhecimento fitoquímico de Cissampelos sympodialis Eichl. (Menispermaceae)
author Lima, Natanael Teles Ramos de
author_facet Lima, Natanael Teles Ramos de
author_role author
dc.contributor.none.fl_str_mv Barbosa Filho, José Maria
http://lattes.cnpq.br/8892459126928726
Fechine, Ivana Maria
http://lattes.cnpq.br/5689715833582093
dc.contributor.author.fl_str_mv Lima, Natanael Teles Ramos de
dc.subject.por.fl_str_mv Milona
Cissampelos
Flavonoides
Docking
Topoisomerase
Citotoxicidade
Flavonoids
Topoisomerase
Cytotoxicity
CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA
topic Milona
Cissampelos
Flavonoides
Docking
Topoisomerase
Citotoxicidade
Flavonoids
Topoisomerase
Cytotoxicity
CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA
description Cissampelos sympodialis Eichl., popularly known as “milona”, “jarrinha”, “abuteira” and “orelha-de-onça”, it is a species rich in alkaloids and has traditional use in the treatment of asthma, bronchitis, rheumatism and arthritis. Thus, the objective of this work was to contribute to the chemical and pharmacological knowledge of the species, through the isolation of substances, mass spectrometry, in silico studies and biological tests. The aerial parts were collected in the garden of UFPB’s Research Institute for Drugs and Medicines – IpeFarM and an exsiccate is deposited in the Herbarium Prof. Lauro Pires Xavier under AGRA 1476 registration and in the National System of Management of Genetic Heritage and Associated Traditional Knowledge (SisGen) under the code A9B2EFC. After harvest, drying and grinding, extraction was performed with 95% ethanol and then the solvent was concentrated in a rotary evaporator, obtaining the Crude Ethanol Extract (CEE). The CEE was partitioned using hexane, chloroform and ethyl acetate. The hydromethanolic phase was subjected to open column chromatography using Sephadex-LH 20, resulting in the CsFHM 6-7 fraction that was processed by preparative High Performance Liquid Chromatography (HPLC). An alkaloids march was performed to obtain the Alkaloid Rich Fraction (ARF). The isolated compounds were identified using Mass spectrometry (MS), Nuclear Magnetic Resonance (NMR) and comparison with the literature. A molecular docking study was carried out on alkaloids already isolated from milona: laurifolin, milonine, warifteina, methylwarifteina, roraimina, des-7'-O-methylroraimina and epi-des-7'-Omethylroraimina; to evaluate their interactions with Topoisomerase IIα and DNA. An MTT assay was performed with samples of C. sympodialis in breast cancer cells, human melanoma, colon carcinoma, cervical adenocarcinoma and in normal human keratinocyte cells. The preparative HPLC allowed the isolation of 4 substances: the nucleoside adenosine and three flavonoids, vicenin-2, quercetin-3-O-βglycopyranosyl-(1→6)-β-galactopyranoside and canferol-3-O-β-glycopyranosyl- (1→6)-β-galactopyranoside. In addition, from the literature, 10 more compounds were identified by LC-MS: caffeic acid-hexoside, salsolinol, guanosine, syringic acid-Ohexoside, magnoflorin, benzyl alcohol-hexose-pentose, aromadendrine-hexoside, apigenin-6-C-glycoside, isoquercetin and astragalin. The docking pointed out that among milona alkaloids, roraimine is the one that best interacts with Toposiomerase IIα and with DNA and des-7'-O-methylroraimine is the best as a DNA ligand. The MTT assay indicated that CEE has inhibitory activity in colon and breast cancer cells and ARF was able to strongly inhibit the growth of all tested strains, although it has an action against normal cells. The test also demonstrated that the glycosylated flavonoids have no activity against the tested cell lines. It is concluded that milona has potential antitumor activity and that it can eventually be used as a complement in the treatment of cancer, in addition to theoretically having chemical compounds that can become active drugs against neoplasms.
publishDate 2020
dc.date.none.fl_str_mv 2020-11-23T16:55:47Z
2020-07-28
2020-11-23T16:55:47Z
2020-02-14
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/123456789/18499
url https://repositorio.ufpb.br/jspui/handle/123456789/18499
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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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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