Determine the enzymatic kinetic characteristics of CYP3A4 variants utilizing artemether-lumefantrine

Artemether-lumefantrine is an artemisinin-based combination therapy for the treatment of malaria, which are primarily metabolized by cytochrome P450 3A4. Therapeutic difference caused by gene polymorphisms of CYP3A4 may lead to uncertain adverse side effects or treatment failure. The aim of this study was to evaluate the effect of CYP3A4 gene polymorphism on artemether-lumefantrine metabolism in vitro. Enzyme kinetics assay was performed using recombinant human CYP3A4 cell microsomes. The analytes, dihydroartimisinin and desbutyl-lumefantrine, were detected by ultra-performance liquid chromatography tandem mass spectrometry. The results demonstrated that compared to CYP3A4.1, the intrinsic clearance of CYP3A4.4, 5, 9, 16, 18, 23, 24, 28, 31–34 significantly reduced for artemether (58.5%–93.3%), and CYP3A4.17 almost loss catalytic activity. Simultaneously, CYP3A4.5, 14, 17, 24 for lumefantrine were decreased by 56.1%–99.6%, and CYP3A4.11, 15, 18, 19, 23, 28, 29, 31–34 for lumefantrine was increased by 51.7%–296%. The variation in clearance rate indicated by molecular docking could be attributed to the disparity in the binding affinity of artemether and lumefantrine with CYP3A4. The data presented here have enriched our understanding of the effect of CYP3A4 gene polymorphism on artemether-lumefantrine metabolizing. These findings serve as a valuable reference and provide insights for guiding the treatment strategy involving artemether-lumefantrine. •A UPLC-MS/MS method was established for the simultaneous detection of artemether-lumefantrine.•Enzyme kinetic parameters for metabolizing artemether-lumefantrine by CYP3A4 variants were obtained.•The variability in the clearance rate of artemether-lumefantrine by CYP3A4 may be related to its affinity.