Molecular Design Strategy to Construct the Near-Infrared Fluorescent Probe for Selectively Sensing Human Cytochrome P450 2J2

• DOI: 10.1021/jacs.8b12136
• Publication Date: 2018-12-10T19:41:22Z
• Authors: Jing Ning; Tao Liu; Peipei Dong; Wei Wang; Guangbo Ge; Bo Wang; Zhenlong Yu; Lei Shi; Xiangge Tian; Xiaokui Huo; Lei Feng; Chao Wang; Chengpeng Sun; Jingnan Cui; Tony D. James; Xiaochi Ma
• Keywords: /dk/atira/pure/subjectarea/asjc/1600/1600; name=General Chemistry; Lymphoma; Mice, Nude; Cytochrome P-450 CYP2J2; 01 natural sciences; /dk/atira/pure/subjectarea/asjc/1300/1303; name=Biochemistry; Cytochrome P-450 Enzyme System; Cell Line, Tumor; /dk/atira/pure/subjectarea/asjc/1500/1503; name=Catalysis; Animals; Humans; /dk/atira/pure/sustainabledevelopmentgoals/good_health_and_well_being; name=SDG 3 - Good Health and Well-being; Fluorescent Dyes; Leukemia; Microscopy, Confocal; Neovascularization, Pathologic; /dk/atira/pure/subjectarea/asjc/1500/1505; name=Colloid and Surface Chemistry; 3. Good health; 0104 chemical sciences; Molecular Docking Simulation; Microscopy, Fluorescence; Xanthenes; Microsomes, Liver; Protein Binding

Cytochrome P450 2J2 (CYP2J2), a key enzyme responsible for oxidative metabolism of various xenobiotics and endogenous compounds, participates in a diverse array of physiological and pathological processes in humans. Its biological role in tumorigenesis and cancer diagnosis remains poorly understood, owing to the lack of molecular tools suitable for real-time monitoring CYP2J2 in complex biological systems. Using molecular design principles, we were able to modify the distance between the catalytic unit and metabolic recognition moiety, allowing us to develop a CYP2J2 selective fluorescent probe using a near-infrared fluorophore ( E)-2-(2-(6-hydroxy-2, 3-dihydro-1 H-xanthen-4-yl)vinyl)-3,3-dimethyl-1-propyl-3 H-indol-1-ium iodide (HXPI). To improve the reactivity and isoform specificity, a self-immolative linker was introduced to the HXPI derivatives in order to better fit the narrow substrate channel of CYP2J2, the modification effectively shortened the spatial distance between the metabolic moiety ( O-alkyl group) and catalytic center of CYP2J2. After screening a panel of O-alkylated HXPI derivatives, BnXPI displayed the best combination of specificity, sensitivity and applicability for detecting CYP2J2 in vitro and in vivo. Upon O-demethylation by CYP2J2, a self-immolative reaction occurred spontaneously via 1,6-elimination of p-hydroxybenzyl resulting in the release of HXPI. Allowing BnXPI to be successfully used to monitor CYP2J2 activity in real-time for various living systems including cells, tumor tissues, and tumor-bearing animals. In summary, our practical strategy could help the development of a highly specific and broadly applicable tool for monitoring CYP2J2, which offers great promise for exploring the biological functions of CYP2J2 in tumorigenesis.