Unraveling the Structural Basis of Selective Inhibition of Human Cytochrome P450 3A5

• Published in: Journal of the American Chemical Society Vol. 143; no. 44; pp. 18467 - 18480
• Main Authors: Wang, Jingheng, Buchman, Cameron D, Seetharaman, Jayaraman, Miller, Darcie J, Huber, Andrew D, Wu, Jing, Chai, Sergio C, Garcia-Maldonado, Efren, Wright, William C, Chenge, Jude, Chen, Taosheng
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 10-11-2021; American Chemical Society (ACS)
• Subjects: Anti-Inflammatory Agents - chemistry; Anti-Inflammatory Agents - pharmacology; Catalytic Domain; Chemistry; Cytochrome P-450 CYP3A - chemistry; Cytochrome P-450 CYP3A - genetics; Cytochrome P-450 CYP3A - metabolism; Cytochrome P-450 CYP3A Inhibitors - chemistry; Cytochrome P-450 CYP3A Inhibitors - pharmacology; Gene Expression Regulation, Enzymologic - drug effects; Humans; Models, Molecular; Protein Conformation; Structure-Activity Relationship
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/jacs.1c07066

The human cytochrome P450 (CYP) CYP3A4 and CYP3A5 enzymes metabolize more than one-half of marketed drugs. They share high structural and substrate similarity and are often studied together as CYP3A4/5. However, CYP3A5 preferentially metabolizes several clinically prescribed drugs, such as tacrolimus. Genetic polymorphism in CYP3A5 makes race-based dosing adjustment of tacrolimus necessary to minimize acute rejection after organ transplantation. Moreover, the differential tissue distribution and expression levels of CYP3A4 and CYP3A5 can aggravate toxicity during treatment. Therefore, selective inhibitors of CYP3A5 are needed to distinguish the role of CYP3A5 from that of CYP3A4 and serve as starting points for potential therapeutic development. To this end, we report the crystal structure of CYP3A5 in complex with a previously reported selective inhibitor, clobetasol propionate (CBZ). This is the first CYP3A5 structure with a type I inhibitor, which along with the previously reported substrate-free and type II inhibitor-bound structures, constitute the main CYP3A5 structural modalities. Supported by structure-guided mutagenesis analyses, the CYP3A5–CBZ structure showed that a unique conformation of the F–F′ loop in CYP3A5 enables selective binding of CBZ to CYP3A5. Several polar interactions, including hydrogen bonds, stabilize the position of CBZ to interact with this unique F–F′ loop conformation. In addition, functional and biophysical assays using CBZ analogs highlight the importance of heme-adjacent moieties for selective CYP3A5 inhibition. Our findings can be used to guide further development of more potent and selective CYP3A5 inhibitors.