Ensartinib (X-396) is a promising
tyrosine kinase inhibitor currently undergoing advanced clinical evaluation for the treatment of
non-small cell lung cancer. In this work, we investigate possible interactions of this promising drug candidate with
ATP-binding cassette (ABC) drug efflux transporters and
cytochrome P450 biotransformation
enzymes (CYPs), which play major roles in multidrug resistance (MDR) and pharmacokinetic drug-drug interactions (DDIs). Accumulation studies showed that
ensartinib is a potent inhibitor of ABCB1 and ABCG2 transporters. Additionally, incubation experiments with recombinant CYPs showed that
ensartinib significantly inhibits
CYP3A4 and
CYP2C9. Subsequent molecular docking studies confirmed these findings.
Drug combination experiments demonstrated that
ensartinib synergistically potentiates the antiproliferative effects of
daunorubicin,
mitoxantrone, and
docetaxel in ABCB1, ABCG2, and CYP3A4-overexpressing cellular models, respectively. Advantageously,
ensartinib's antitumor efficiency was not compromised by the presence of MDR-associated
ABC transporters, although it acted as a substrate of ABCB1 in Madin-Darby Canine Kidney II (MDCKII) monolayer transport assays. Finally, we demonstrated that
ensartinib had no significant effect on the
mRNA-level expression of examined transporters and
enzymes in physiological and lung
tumor cellular models. In conclusion,
ensartinib may perpetrate clinically relevant pharmacokinetic DDIs and modulate ABCB1-, ABCG2-, and CYP3A4-mediated MDR. The in vitro findings presented here will provide a valuable foundation for future in vivo investigations.