2. Academic Validation
  3. Rationally Engineered CYP3A4 Fluorogenic Substrates for Functional Imaging Analysis and Drug-Drug Interaction Studies

Rationally Engineered CYP3A4 Fluorogenic Substrates for Functional Imaging Analysis and Drug-Drug Interaction Studies

  • J Med Chem. 2023 May 25;66(10):6743-6755. doi: 10.1021/acs.jmedchem.3c00101.
Rong-Jing He 1,  Zhen-Hao Tian 2,  Jian Huang 3,  Meng-Ru Sun 1,  Feng Wei 4,  Chun-Yu Li 1,  Hai-Rong Zeng 1,  Feng Zhang 1,  Xiao-Qing Guan 1,  Yan Feng 4,  Xiang-Ming Meng 4 5,  Hui Yang 2,  Guang-Bo Ge 1
Affiliations

Affiliations

  • 1 Shanghai Frontiers Science Center of TCM Chemical Biology, Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
  • 2 School of Life Sciences, Northwestern Polytechnical University, Xi'an, Shaanxi 710072, China.
  • 3 Pharmacology and Toxicology Division, Shanghai Institute of Food and Drug Control, Shanghai 201203, China.
  • 4 School of Chemistry and Chemical Engineering & Center for Atomic Engineering of Advanced Materials & Anhui Province Key Laboratory of Chemistry for Inorganic/Organic Hybrid Functionalized Materials, Anhui University, Hefei 230601, China.
  • 5 Institute of Physical Science and Information Technology, Anhui University, Hefei 230601, China.
PMID: 37145039 DOI: 10.1021/acs.jmedchem.3c00101
Abstract

Cytochrome P450 3A4 (CYP3A4) is a key xenobiotic-metabolizing enzyme-mediated drug metabolism and drug-drug interaction (DDI). Herein, an effective strategy was used to rationally construct a practical two-photon fluorogenic substrate for hCYP3A4. Following two-round structure-based substrate discovery and optimization, we have successfully constructed a hCYP3A4 fluorogenic substrate (F8) with desirable features, including high binding affinity, rapid response, excellent isoform specificity, and low cytotoxicity. Under physiological conditions, F8 is readily metabolized by hCYP3A4 to form a brightly fluorescent product (4-OH F8) that can be easily detected by various fluorescence devices. The practicality of F8 for real-time sensing and functional imaging of hCYP3A4 has been examined in tissue preparations, living cells, and organ slices. F8 also demonstrates good performance for high-throughput screening of hCYP3A4 inhibitors and assessing DDI potentials in vivo. Collectively, this study develops an advanced molecular tool for sensing CYP3A4 activities in biological systems, which strongly facilitates CYP3A4-associated fundamental and applied research studies.

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