IntroductionVorolanib is a new-generation multi-targeted kinase inhibitor (MTKI) whose drug-drug interaction (DDI) profile remains largely unexplored. This study combined in vitro and in vivo approaches to systematically investigate the effect of resveratrol on vorolanib metabolism.MethodsUsing ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), the concentrations of vorolanib and its primary active metabolite X297 were quantified in rat liver microsomes (RLM), human liver microsomes (HLM), recombinant human CYP3A4 (rCYP3A4), and Sprague-Dawley rats. Molecular docking simulations were performed to investigate the binding poses of vorolanib and resveratrol within the active site of CYP3A4.ResultsIn vitro assays revealed that resveratrol inhibited vorolanib metabolism with half-maximal inhibitory concentration (IC50) values of 6.28 ± 0.27, 33.86 ± 0.65 and 6.70 ± 0.32 μM in RLM, HLM and rCYP3A4, respectively. The specific mechanisms were identified as non-competitive inhibition in HLM, and mixed inhibition in both RLM and rCYP3A4. IC50 shift experiments indicated that the inhibition of vorolanib by resveratrol was non-time-dependent. Furthermore, metabolic stability assays in RLM showed that resveratrol substantially prolonged the in vitro t1/2 of vorolanib from 39.02 ± 2.02 to 186.24 ± 58.99 min. Molecular docking confirmed that both compounds bind within the catalytic pocket of CYP3A4 with binding energies of −7.29 and −7.23 kcal/mol, respectively. The in vivo pharmacokinetic study demonstrated that co-administration with resveratrol significantly increased the AUC(0-t) and AUC0‐∞ of vorolanib by 1.70- and 1.82-fold, respectively, while significantly decreasing its CLz/F by 56.1%. Similarly, for the metabolite X297, the AUC(0-t), AUC0‐∞, and Cmax were increased by 2.00-, 2.22-, and 1.18-fold, respectively, accompanied by a 63.9% decrease in CLz/F.ConclusionBoth in vitro and in vivo results consistently demonstrate that resveratrol profoundly inhibits the metabolism and elimination of vorolanib, driven by its interference with the CYP3A4 catalytic domain as corroborated by docking analysis. These findings highlight a potential risk of herb-drug interactions, suggesting that close clinical monitoring for adverse reactions is required when vorolanib and resveratrol are used in combination.