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Carolina Oliveira Avaliação Da Atividade Biológica Dos Extratos De Plectranthus Mutabilis, P

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Carolina Oliveira Avaliação Da Atividade Biológica Dos Extratos De Plectranthus Mutabilis, P CAROLINA ISABEL FAUSTINO PEREIRA DE OLIVEIRA AVALIAÇÃO DA ATIVIDADE BIOLÓGICA DOS EXTRATOS DE PLECTRANTHUS MUTABILIS, P. CILIATUS, P. XEROPHILUS, P. CRASSUS E P. LUCIDUS Orientadora: Professora Doutora Patrícia Dias Mendonça Rijo Universidade Lusófona de Humanidades e Tecnologias Escola de Ciências e Tecnologias da Saúde Lisboa 2019 CAROLINA ISABEL FAUSTINO PEREIRA DE OLIVEIRA AVALIAÇÃO DA ATIVIDADE BIOLÓGICA DOS EXTRATOS DE PLECTRANTHUS MUTABILIS, P. CILIATUS, P. XEROPHILUS, P. CRASSUS E P. LUCIDUS Dissertação defendida em provas públicas na Universidade Lusófona de Humanidades e Tecnologias no dia 24/04/2020, perante o júri, nomeado pelo Despacho de Nomeação nº: 42/2020, de 12 de Fevereiro de 2020, com a seguinte composição: Presidente: Prof. Doutor Luís Monteiro Rodrigues Arguente: Profª. Doutora Mariana Reis Orientadora: Profª. Doutora Patrícia Dias Mendonça Rijo Universidade Lusófona de Humanidades e Tecnologias Escola de Ciências e Tecnologias da Saúde Lisboa 2019 “Determinação, coragem e autoconfiança são fatores decisivos para o sucesso. Não importa quais sejam os obstáculos e as dificuldades, se estamos possuídos de uma inabalável determinação, conseguiremos superá-los. Independentemente das circunstâncias, devemos ser sempre humildes, recatados e despidos de orgulho.” Dalai Lama Agradecimentos Este trabalho foi desenvolvido no âmbito da unidade curricular de Projeto e, como tal, o meu primeiro agradecimento vai para a Professora Doutora Patrícia Dias Mendonça Rijo que, há três anos, me tem vindo a orientar com empenho e a incentivar à participação em eventos de grande importância para a investigação. Agradeço à Escola de Ciências e Tecnologias da Saúde (ECTS) da Universidade Lusófona, pelo reconhecimento do meu trajeto curricular e consequente atribuição da bolsa PADDIC (Programa de apoio ao desenvolvimento e divulgação da investigação científica), que me incentivou na realização deste projeto. Agradeço à Professora Doutora Catarina Garcia e Professora Epole Ntungwe da ECTS, da Universidade Lusófona, por tornarem possível a realização deste trabalho e pela amizade. Agradeço à Professora Doutora Lucília Saraiva do Laboratório de Microbiologia, do Departamento de Ciências Biológicas da Faculdade de Farmácia da Universidade do Porto pela colaboração na realização de ensaios de citotoxicidade em células tumorais humanas. Agradeço às minhas colegas e amigas Catarina Teodósio, Cláudia Oliveira e Raquel Ferreira, pelo companheirismo e amizade ao longo destes cinco anos e que levo para a vida. Agradeço aos meus amigos de infância Mónica Luís, João Guisado, Ana Belchior, Guilherme Teixeira, Ana Soares e, namorado, Diogo Rosa por serem o meu escape sempre que os obstáculos pareciam intransponíveis. E por último, mas não menos importante, agradeço a toda a minha familia, em particular: país, Paula Oliveira e Jorge Oliveira, por serem os primeiros a acreditarem em mim e pelo apoio incondicional; avós, Isabel Ilhéu e Raul Ilhéu, por terem sido os meus pilares quando mais precisei; irmão, João Oliveira, pelo exemplo de vida que foi para mim e que, apesar de hoje estar apenas presente no meu coração, foi essencial para que eu me tornasse na pessoa que sou hoje e prima, Adriana Oliveira, pela disponibilidade para me aconselhar e ajudar sempre que precisei. A todos expresso um enorme obrigada. Carolina Oliveira Avaliação da atividade biológica dos extratos de Plectranthus mutabilis, P. ciliatus, P. xerophilus, P. crassus e P. lucidus Produção Científica Os resultados presentes nesta dissertação bem como outros trabalhos realizados encontram-se identificados abaixo. Artigos científicos - Ntungwe, E., Marçalo, J., Garcia, C., Reis, C., Teodósio, C., Oliveira, C. I., Oliveira, C. F., Roberto, A. & Rijo, P. (2017). Biological activity screening of seven Plectranthus species. Journal Biomedical and Biopharmaceutical Research, 14(1), 95–108. https://doi.org/10.19277/bbr.14.1.153. - Ntungwe, E., Bessa, C., Garcia, C., Teodósio, C., Oliveira, C., Oliveira, C., Andrade, J., Isca, V., Roberto, A., Saraiva, L. & Rijo, P. (2018). 7-Acetoxy-6-hydroxyroyleanone from Plectranthus ramosior: a screening study for anticancer lead molecules. Planta medica. Em submissão. - Garcia, C., Teodósio, C., Oliveira, C., Oliveira, C., Diaz-Lanza, A., Reis, C., Duarte, N. & Rijo, P. (2018). Naturally occurring Plectranthus-derived diterpenes with antitumoral activities. Current Pharmaceutical Design, Volume 24, Number 36, 2018, pp. 4207-4236(30). https://doi.org/10.2174/1381612825666190115144241. - Macedo, A., Filipe, P., Thomé, N. G., Vieira, J., Oliveira, C., Teodósio, C., Ferreira, R., Roque, L. & Fonte, P. (2020). A Brief Overview of the Oral Delivery of Insulin as an Alternative to the Parenteral Delivery. Current Molecular Medicine, 20, 1–10. https://doi.org/10.2174/1566524019666191010095522 Comunicações em painel nacionais - Carolina Oliveira, Catarina Teodósio, Cláudia Oliveira, Marisa Nicolai, Paula Pereira, Amilcar Roberto, Patrícia Rijo, “Antioxidant activity and rosmarinic acid content of ultrasound-assisted ethanolic extracts of medicinal plants”; 13º Congresso das Fármacias da Associação Nacional das Farmácias (ANF), 19 outubro, Lisboa, Portugal, 2018. - Catarina Teodósio, Carolina Oliveira, Cláudia Oliveira, Ntungwe Epole, Joana M. Andrade, Vera Isca, Patrícia Rijo, “Plectranthus species as a source of anticancer lead molecules”; 13º Congresso das Farmácias da Associação Nacional das Farmácias (ANF), 19 outubro, Lisboa, Portugal, 2018. 6 Universidade Lusófona de Humanidades e Tecnologias, Escola de Ciências e Tecnologias da Saúde Carolina Oliveira Avaliação da atividade biológica dos extratos de Plectranthus mutabilis, P. ciliatus, P. xerophilus, P. crassus e P. lucidus - C. Garcia, C. Oliveira, C.S. Oliveira, A. Roberto, C. Reis, M. Nicolai, P. Rijo, “Determination of the antioxidante activity of medicinal plants extracts – a didactic approach”; II Jornadas CBIOS, 12 outubro, Lisboa, Portugal, 2016. - Epole Ntungwe N, Carolina Oliveira, Catarina Teodósio, Vera Isca, Patrícia Rijo, “Bioguided fractionation and isolation study of bioactive componentes from Plectranthus mutabilis Codd”; 1ª Conferência sobre investigação em Produtos Naturais e Avícolas- Iniciativa conjunta CBIOS-CYTED, 8 e 9 novembro, Lisboa, Portugal, 2018. Comunicações em painel internacionais - Henrique Silva, Ntungwe Epole, Joana Marçalo, Catarina Garcia, Catarina Reis, Catarina Teodósio, Carolina Oliveira, Cláudia Oliveira, Luís Rodrigues, Patrícia Rijo, “Estimation of acetylcholinesterase inhibition, antioxidant, antimicrobial activities and brine shrimp, toxicity of seven Plectranthus species”; International Federation Congress (FIP), Estocolmo, Suécia, 2017. - Epole Ntungwe N, Joana Marçalo, Catarina Garcia, Catarina Reis, Catarina Teodósio, Carolina Oliveira, Cláudia Oliveira, Amílcar Roberto, Patrícia Rijo, “Biological activity screening of seven Plectranthus species: anticancer lead molecule search” (P5); 2nd Training School COST Action CM 1407, 18-20 setembro, Lisboa, Portugal, 2017. Proceeding nacionail - C. Garcia, C. Oliveira, C.S. Oliveira, A. Roberto, C. Reis, M. Nicolai, P. Rijo, “Determination of the antioxidante activity of medicinal plants extracts – a didactic approach”, Biomed Biopharm Res., 2016; (P9) 2 :, 273-290. Proceeding internacional - Epole Ntungwe N, Joana Marçalo, Catarina Garcia, Catarina Reis, Catarina Teodósio, Carolina Oliveira, Cláudia Oliveira, Amílcar Roberto, Patrícia Rijo, “Biological activity screening of seven Plectranthus species: anticancer lead molecule search” (P10); 4th meeting of COST Action CM 1407, 21-22 setembro, Lisboa, Portugal, 2017. 7 Universidade Lusófona de Humanidades e Tecnologias, Escola de Ciências e Tecnologias da Saúde Carolina Oliveira Avaliação da atividade biológica dos extratos de Plectranthus mutabilis, P. ciliatus, P. xerophilus, P. crassus e P. lucidus Suplemento - E. Ntungwe, J. Marçalo, C. Garcia, C. Reis, C. Teodósio, C. Oliveira, C. F. Oliveira, A. Roberto, P. Rijo, “Biological activity screening of seven Plectranthus species:Anticancer lead molecule search”, Biomed Biopharm Res., 2017; (P5) 2:, 253-286. https://doi.org/10.19277/bbr.14.2.168. 8 Universidade Lusófona de Humanidades e Tecnologias, Escola de Ciências e Tecnologias da Saúde Carolina Oliveira Avaliação da atividade biológica dos extratos de Plectranthus mutabilis, P. ciliatus, P. xerophilus, P. crassus e P. lucidus Resumo Neste trabalho foi avaliada a atividade biológica de cinco espécies do género Plectranthus (P. mutabilis, P. lucidus, P. xerophilus, P. crassus e P. ciliatus). As bioatividades testadas foram as atividades antioxidante, pelo método de DPPH, e antimicrobiana, pelo método de difusão em poços, e a toxicidade geral, pelo modelo de Artemia salina L. O extrato de P. mutabilis apresentou o maior rendimento de extração (30,3%) e a atividade antioxidante mais elevada (46,14%). Demonstrou atividade antimicrobiana significativa contra S. aureus (17 mm) e moderada contra C. albicans (10 mm) pelo que, posteriormente, determinaram-se os valores da CMI (31,25 e 125 g/mL, respetivamente), CMB e CMF (125 g/mL), pelo método de microdiluição. Esta planta demonstrou também a toxicidade geral mais elevada (CL50 = 0,55 mg/L) e, portanto, a sua citotoxicidade foi testada em três linhas celulares tumorais humanas (HCT116 = 28.00±2.00; MCF-7 = 35.00±1.00 e NCI-H460 = 36.00±2.00 µg/mL). O extrato de P. mutabilis foi o mais bioativo e foi selecionado para a realização de técnicas cromatográficas de fracionamento e identificação estrutural por espetroscopia
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