Premium
In vitro metabolism of the lignan (−)‐grandisin, an anticancer drug candidate, by human liver microsomes
Author(s) -
Barth Thiago,
Habenschus Maísa Daniela,
Lima Moreira Fernanda,
Ferreira Leandro De Santis,
Lopes Norberto Peporine,
Moraes de Oliveira Anderson Rodrigo
Publication year - 2015
Publication title -
drug testing and analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.065
H-Index - 54
eISSN - 1942-7611
pISSN - 1942-7603
DOI - 10.1002/dta.1743
Subject(s) - metabolite , chemistry , lignan , biotransformation , chromatography , high performance liquid chromatography , microsome , mass spectrometry , drug metabolism , metabolism , biochemistry , in vitro , stereochemistry , enzyme
(−)‐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 (V max ) was 3.96 ± 0.18 µmol/mg protein/h, and the Michaelis‐Menten constant (K m ) 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. Copyright © 2015 John Wiley & Sons, Ltd.