Avaliação do potencial antitripanossoma de neolignanas modificadas via metátese olefínica cruzada obtidas de partes aéreas de Nectandra leucantha Nees & Mart. (Lauraceae)
Data
2023-11-28
Autores
Ueno, Anderson Keity 
Orientadores
Lago, João Henrique Ghilardi
Tipo
Tese de doutorado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
doença de Chagas, cujo agente etiológico é o parasita Trypanossoma cruzi, faz parte do rol de moléstias categorizadas como doenças tropicais negligenciadas (DTN) por ter ainda recursos terapêuticos limitados para o seu tratamento e cerca de 6 milhões de pessoas atingidas no Brasil. A fitoquímica, diante deste cenário, é uma área promissora para a busca de substâncias bioativas com atividades antiparasitárias. A espécie Nectandra leucantha (Ness & Mart) apresenta como principais marcadores neolignanas do tipo dehidrodieugenol, as quais possuem as atividades citadas. A substância dehidrodieugenol B (1) foi obtida a partir do fracionamento cromatográfico do extrato hexânico das folhas e galhos de N. leucantha e o respectivo derivado metilado metildehidrodieugenol B (2) foi preparado atavés da metilação de (1). Estes compostos foram utilizados como materiais de partida para o preparo de oito derivado esterificados (1(a-d) e 2(a-d)) os quais foram obtidos através de protocolo de metátese olefínica cruzada e purificados através de técnicas cromatográficas. As respectivas identificações foram feitas através de métodos espectroscópicos (Infravermelho e Ressonância Magnética Nuclear) e espectrométricos (Espectrometria de Massas). Os derivados obtidos foram avaliados para atividade antitripanosoma frente as formas tripomastigota e amastigota de Trypanossoma cruzi em ensaios in vitro. Os compostos 1a, 1b, 1c, 1d e 2c apresentaram valores de CE50 frente a tripomastigota respectivamente de: 13,5 μM, 1,51 μM, 0,68 μM, 3,89 μM e 2,70 μM. Estes valores foram inferiores ao padrão benznidazol (BZD 18,7 μM). Os compostos que apresentaram valores inferiores ao controle BZD (5,5 μM) frente a amastigota foram: 1b (2,68 μM), 1c(3,49 μM) e 1d (2,09 μM). A citotoxicidade dos compostos foi avaliada em ensaios in vitro empregando células NCTC. Os compostos 2a, 2b e 2d apresentaram valores de CC50 superiores a 200 μM enquanto que 1b, 1c, 1d e 2c apresentaram valores inferiores a 100 μM. O composto 2a representa o melhor canditado entre os protótipos preparados por ter potência elevada, não citotoxicidade e seletividade (IS >32,8). Além disto, 2a apresentou valor de logP de 4,43, boa absorção pelo trato gastrointestinal e hidrossolubilidade em predições in silico apresentando condições mínimas para formulação oral.
Chagas disease, whose etiological agent is the parasite Trypanosoma cruzi, is part of the group of diseases categorized as neglected tropical diseases (NTDs) due to its limited therapeutic resources for treatment and approximately 6 million affected individuals in Brazil. In this scenario, phytochemistry emerges as a promising field for searching bioactive substances with anti-parasitic activities. The species Nectandra leucantha (Ness & Mart) presents as its main phytochemical marker dehydrodieugenol neolignan´s type, which possess the aforementioned activities. The primary objective of this study was to evaluate the antritrypanossome activity against both trypomastigote and amastigote forms of Trypanosoma cruzi employing semi-synthetic derivatives (1a-1d) and (2a-2d) generated through cross-olefin metathesis procedure. These derivatives were synthesized from neolignans Dehydrodieugenol B (1) and Methyldehydrodieugenol B (2), isolated from the aerial parts of Nectandra leucantha (Ness & Mart). The isolation of neolignans 1 and 2 was achieved through chromatographic techniques of hexanic extract from the aerial parts of N. leucantha. Structural characterization of the semi-syntethic derivatives 1(a-d) and 2(a-2d) was conducted through spectroscopic techniques (Infrared, Nuclear Magnetic Resonance) as well as spectrometric analysis (Mass Spectrometry). Subsequently, the compounds were subjected to in vitro assays to evaluate their potencial against trypomastigotes and amastigotes of T. cruzi. Among the compounds tested 1a, 1b, 1c, 1d and 2c displayed EC50 values against trypomastigotes of 13,5μM, 1,5 μM, 0,7 μM, and 3,9 μM, respectively, which were lower than the reference benznidazole (BZD – 18,7 μM). For amastigotes, the compounds that displayed values lower than the BZD control (5,5 μM) were 1b (2,7 μM), 1c (3,5 μM), and 1d (2,1 μM). The cytotoxicity of these compounds was evaluated by in vitro using NCTC cells, revealing that 2a, 2b, and 2d exhibited CC50 values greater than 200 μM, while 1b, 1c, 1d, and 2c demonstrated values below 100 μM. Compound 2a is most suitable among others due to its high potency, lack of cytotoxicity, and selectivity (IS > 32.8). Additionally, 2a exhibited a logP value of 4.43, good gastrointestinal absorption, and hydrophilicity predicted by in silico, indicating conditions for oral formulation.
Chagas disease, whose etiological agent is the parasite Trypanosoma cruzi, is part of the group of diseases categorized as neglected tropical diseases (NTDs) due to its limited therapeutic resources for treatment and approximately 6 million affected individuals in Brazil. In this scenario, phytochemistry emerges as a promising field for searching bioactive substances with anti-parasitic activities. The species Nectandra leucantha (Ness & Mart) presents as its main phytochemical marker dehydrodieugenol neolignan´s type, which possess the aforementioned activities. The primary objective of this study was to evaluate the antritrypanossome activity against both trypomastigote and amastigote forms of Trypanosoma cruzi employing semi-synthetic derivatives (1a-1d) and (2a-2d) generated through cross-olefin metathesis procedure. These derivatives were synthesized from neolignans Dehydrodieugenol B (1) and Methyldehydrodieugenol B (2), isolated from the aerial parts of Nectandra leucantha (Ness & Mart). The isolation of neolignans 1 and 2 was achieved through chromatographic techniques of hexanic extract from the aerial parts of N. leucantha. Structural characterization of the semi-syntethic derivatives 1(a-d) and 2(a-2d) was conducted through spectroscopic techniques (Infrared, Nuclear Magnetic Resonance) as well as spectrometric analysis (Mass Spectrometry). Subsequently, the compounds were subjected to in vitro assays to evaluate their potencial against trypomastigotes and amastigotes of T. cruzi. Among the compounds tested 1a, 1b, 1c, 1d and 2c displayed EC50 values against trypomastigotes of 13,5μM, 1,5 μM, 0,7 μM, and 3,9 μM, respectively, which were lower than the reference benznidazole (BZD – 18,7 μM). For amastigotes, the compounds that displayed values lower than the BZD control (5,5 μM) were 1b (2,7 μM), 1c (3,5 μM), and 1d (2,1 μM). The cytotoxicity of these compounds was evaluated by in vitro using NCTC cells, revealing that 2a, 2b, and 2d exhibited CC50 values greater than 200 μM, while 1b, 1c, 1d, and 2c demonstrated values below 100 μM. Compound 2a is most suitable among others due to its high potency, lack of cytotoxicity, and selectivity (IS > 32.8). Additionally, 2a exhibited a logP value of 4.43, good gastrointestinal absorption, and hydrophilicity predicted by in silico, indicating conditions for oral formulation.
Descrição
Citação
UENO, Anderson K. Avaliação do potencial antitripanossoma de neolignanas modificadas via metátese olefínica cruzada obtidas de partes aéreas de Nectandra leucantha Nees & Mart. (Lauraceae). 2023. 207 f. Tese (Doutorado em Biologia Química) - Instituto de Ciências Ambientais, Químicas e Farmacêuticas, Universidade Federal de São Paulo, Diadema, 2023. Paulo