@article{e759009954644da8a4e8bad37aced9f3,
title = "Elucidating the binding and metabolic interactions of sunitinib and sorafenib with Cytochrome P450s CYP2U1 and CYP2D6",
abstract = "Cytochrome P450 2U1 (CYP2U1) is an extrahepatic monooxygenase that metabolizes both endogenous fatty acids and xenobiotic substrates. Because of its high expression in both healthy and cancerous thymus tissues, this study investigated CYP2U1-mediated biotransformation of 2 tumor-targeting tyrosine kinase inhibitors, sorafenib and sunitinib, both are commonly prescribed for thymus cancers. Recombinantly expressed CYP2U1 and CYP2D6 were incorporated into nanodiscs, and their metabolism of sorafenib and sunitinib was investigated. Liquid chromatography–tandem mass spectrometry metabolite profiling revealed that both CYP2U1 and CYP2D6 nanodiscs oxidized sorafenib to generate sorafenib N-oxide and both enzymes catalyzed the dealkylation of sunitinib to form N-desethyl sunitinib. Spectroscopic studies (UV-Visible and fluorescence) confirmed favorable binding interactions of CYP2U1 and CYP2D6 to both drugs tested. Molecular dynamics simulations demonstrated binding of sorafenib and sunitinib in the CYP2U1 active site and identified key interactions between the drug and key residues at the enzyme{\textquoteright}s active site. Rates of metabolite formation were quantified by targeted metabolomics, and inhibition of tyrosine kinase was assessed by ELISA assay. Both the parent compound sorafenib and metabolite sorafenib N-oxide showed similar inhibitory effects on cellular migration in HepG2 cells. The metabolite sorafenib N-oxide was approximately twice as potent as the parent compound in inhibiting cancer cell migration. In contrast, N-desethyl sunitinib failed to show similar extent of inhibition. Together, these findings highlight the potential role of extrahepatic CYP2U1 in the local metabolism of tyrosine kinase inhibitors and suggest that CYP2U1-mediated transformations directly influence antitumor efficacy at thymic tumor sites.Significance StatementUnderstanding the interactions between cytochrome P450 2U1 and cytochrome P450 2D6 in nanodiscs and thymus tumor-targeting drugs, sorafenib and sunitinib, led to discovery of new bioactive metabolites that carry differential anticancer properties compared with their parent compounds.",
keywords = "Cytochrome P450 2D6, Cytochrome P450 2U1, Drug metabolism, Thymus cancer, Tyrosine kinase inhibition",
author = "Tang, \{Tiffany Y.C.\} and Yupeng Li and Peter Chapman and Emad Tajkhorshid and Aditi Das",
note = "We would like to thank Jean-Luc Boucher (Universit{\'e} Paris Descartes, Paris Sorbonne Cit{\'e}, Paris, France) and Eric Johnson (Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California) for the N-terminus modified CYP2U1 and CYP2D6 gene constructs. We also thank F. Peter Guengerich for full-length CYP2U1 mammalian gene construct and relevant discussion. We would like to acknowledge Lucas Li from the Duke Proteomics Center for method development on the targeted mass spectrometry, and Sam Moore from the mass spectrometry core of Georgia Tech for untargeted mass spectrometry sample processing.DisclaimerThe content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.Financial supportThe financial support was provided by theNational Institutes of Health Maximizing Investigators{\textquoteright} ResearchAward [GrantR35GM152121] (to T.Y.C.T, P.C., and A.D.),Air Force Office of Scientific Research[GrantAWD005464] (to T.Y.C.T and A.D.),National Institutes of Health[GrantR24-GM145965] (to Y.L. and E.T.). This research was also funded in part by theBeckman Institute for Advanced Scienceand Technology Graduate Fellows Program with support from theArnold and Mabel Beckman Foundation(Y.L.). The molecular dynamics simulations were performed using the computational resources provided by the NSF Supercomputing Centers [ACCESS Grant MCA06N060].Data availabilityThis article contains no datasets generated or analyzed during the current study.CRediT authorship contribution statementTiffany Y.-C. Tang: Conceptualization, Formal analyses, Investigation, Validation, Writing – original draft. Yupeng Li: Conceptualization, Formal analyses, Investigation, Writing – original draft. Peter Chapman: Investigation. Emad Tajkhorshid: Conceptualization, Methodology, Supervision, Funding acquisition. Aditi Das: Conceptualization, Methodology, Supervision, Writing – review and editing, Funding acquisition. The financial support was provided by the National Institutes of Health Maximizing Investigators{\textquoteright} Research Award [Grant R35GM152121 ] (to T.Y.C.T, P.C., and A.D.), Air Force Office of Scientific Research [Grant AWD005464 ] (to T.Y.C.T and A.D.), National Institutes of Health [Grant R24-GM145965 ] (to Y.L. and E.T.). This research was also funded in part by the Beckman Institute for Advanced Science and Technology Graduate Fellows Program with support from the Arnold and Mabel Beckman Foundation (Y.L.). The molecular dynamics simulations were performed using the computational resources provided by the NSF Supercomputing Centers [ACCESS Grant MCA06N060].",
year = "2026",
month = apr,
doi = "10.1016/j.molpha.2026.100114",
language = "English (US)",
volume = "108",
journal = "Molecular Pharmacology",
issn = "0026-895X",
publisher = "Elsevier Inc.",
number = "4",
}