Effects of Agrimonolide on Human Cytochrome P450 Isoforms Using Recombinant Enzymes and HepaRG Cells

Cytochrome P450 (CYP) plays a crucial role in the metabolism of small molecules, primarily in the liver and extrahepatic tissues.The inhibition and/or induction of CYP by exogenous substances is often associated with clinically significant herb-drug interactions (HDIs).HDIs are mediated by co-administered drugs and herbs, which can increase the complexity of treatment, leading to drug adverse effects or reduced drug efficacy.Numerous natural compounds have been studied for their potential to induce or inhibit CYP activity.In this study, we selected agrimonolide (AM), extracted from Agrimonia pilosa, to evaluate its effects on CYP activity.First, the inhibitory effects of AM on CYP reactions (CYP1A2, CYP2B6, CYP2C9, CYP2C19, CYP2D6, and CYP3A4) were investigated in recombinant enzymes to assess potential HDIs.The results confirmed that AM inhibited CYP isoforms, with moderate effects on CYP1A2 and CYP3A4, and weak effects on the remaining isoforms.In particular, these inhibitory effects were in the form of competitive and mixed-type inhibition in CYP1A2 and CYP3A4, respectively.Lineweaver-Burk plots and molecular docking studies further confirmed that AM binds to the enzyme active site.In HepaRG cells, however, CYP1A2 and CYP3A4 were significantly inhibited only when AM was administered at higher concentrations.Predicting CYP-specific binding in the early stages of drug development with natural compounds is essential, as information regarding CYP interactions can help facilitate drug modification and assess the potential for HDIs.Our study suggests that AM, in combination with herbal preparations, may affect drug metabolism by modulating CYP enzyme activity.