Background: Thegene polymorphisms of the CYP2C9, as well as the substrate specificity of theenzyme, result in different clearances for different substrates by CYP2C9variants.
Research designand methods: The CYP2C9 wild type and 38 CYP2C9 variants, expressed in insectmicrosomes, were incubated with azilsartan. The resulting metabolite,O-desethyl azilsartan, was determined by HPLC-MS/MS. The enzyme kineticparameters of the 38 variants were calculated and compared with the wild type.Subsequently, we selected CYP2C9 × 1, *2, and * 3 as target proteins for moleculardocking with azilsartan to elucidate the mechanisms underlying changes inenzyme function.
Results: Comparedwith CYP2C9 × 1, three variants (CYP2C9 × 29, *39, and * 49) exhibited markedlyincreased CLint values (from 170%-275%, *p < 0.05), whereas 28variants exhibited significantly decreased CLint values (from3%-63%,*p < 0.05). The molecular docking results showed that the binding energy ofCYP2C9 × 2 and * 3 was lower than that of the wild type.
Conclusion: Thisassessment revealed the effect of CYP2C9 gene polymorphisms on azilsartanmetabolism, establishing a theoretical basis for further in-vivo studies andclinical applications. This study will help expand the database of CYP2C9gene-drug pairs and identify appropriate treatment strategies for azilsartan,contributing to the field of precision medicine.
Keywords: Azilsartan; CYP2C9; Genetic polymorphism; HPLC-MS/MS; metabolism in vitro.