title: In vitro metabolism of the lignan (-)-grandisin, an anticancer drug candidate, by human liver microsomes.
abstract: (-)-grandisin is a tetrahydrofuran lignan that displays important biological properties, such as trypanocidal, anti-inflammatory, cytotoxic, and antitumor activities, suggesting its utility as a potential drug candidate. One important step in drug development is metabolic characterization and metabolite identification. To perform a biotransformation study of (-)-grandisin and to determine its kinetic properties in humans, a high performance liquid chromatography (HPLC) method was developed and validated. After HPLC method validation, the kinetic properties of (-)-grandisin were determined. (-)-grandisin metabolism obeyed Michaelis-Menten kinetics. The maximal reaction rate (Vmax ) was 3.96 ± 0.18 µmol/mg protein/h, and the Michaelis-Menten constant (Km ) was 8.23 ± 0.99 μM. In addition, the structures of the metabolites derived from (-)-grandisin were characterized via gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) analysis. Four metabolites, 4-O-demethylgrandisin, 3-O-demethylgrandisin, 4,4'-di-O-demethylgrandisin, and a metabolite that may correspond to either 3,4-di-O-demethylgrandisin or 3,5-di-O-demethylgrandisin, were detected. CYP2C9 isoform was the main responsible for the formation of the metabolites. These metabolites have not been previously described, demonstrating the necessity of assessing (-)-grandisin metabolism using human-derived materials.
authors.affiliation: Núcleo de Pesquisa em Produtos Naturais e Sintéticos (NPPNS), Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, 14040-903, Ribeirão Preto, SP, Brazil.|Curso de Farmácia, Universidade Federal do Rio de Janeiro, 27930-560, Macaé-RJ, Brazil.
authors.forename: Thiago
authors.initials: T
authors.lastname: Barth
authors.affiliation: Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, 14040-901, Ribeirão Preto-SP, Brazil.
authors.forename: Maísa Daniela
authors.initials: MD
authors.lastname: Habenschus
authors.affiliation: Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, 14040-903, Ribeirão Preto, São Paulo, Brazil.
authors.forename: Fernanda
authors.initials: F
authors.lastname: Lima Moreira
authors.affiliation: Lychnoflora Pesquisa & Desenvolvimento em Produtos Naturais LTDA, Rua Ângelo Mestriner 263, 14030-090, Vila Virgínia, Ribeirão Preto-SP, Brazil.
authors.forename: Leandro De Santis
authors.initials: Lde S
authors.lastname: Ferreira
authors.affiliation: Núcleo de Pesquisa em Produtos Naturais e Sintéticos (NPPNS), Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, 14040-903, Ribeirão Preto, SP, Brazil.
authors.forename: Norberto Peporine
authors.initials: NP
authors.lastname: Lopes
authors.affiliation: Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, 14040-901, Ribeirão Preto-SP, Brazil.
authors.forename: Anderson Rodrigo
authors.initials: AR
authors.lastname: Moraes de Oliveira
mesh_terms: D000970:Antineoplastic Agents / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D005663:Furans / Q000378:metabolism*; D006801:Humans; D017705:Lignans / Q000378:metabolism*; D008862:Microsomes, Liver / Q000378:metabolism*; D020033:Protein Isoforms / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D005663:Furans; D017705:Lignans; D020033:Protein Isoforms; C074346:grandisin; D003577:Cytochrome P-450 Enzyme System
keywords: (−)-grandisin; human liver microsomes; in vitro metabolism; metabolite identification