Vicagrel is hydrolyzed by Raf kinase inhibitor protein in human intestine
Vicagrel is hydrolyzed by Raf kinase inhibitor protein in human intestine
复制标题
DOI：
10.1002/bdd.2340
复制
发表时间：
2022-12
影响因子：
2.1
通讯作者：
Ting Zhu;Yu Wu;Xue-Mei Li;Yu-Meng Jia;Huan Zhou;Li jiang;T. Tai;Qiong-Yu Mi;Jin-Zi Ji;Hong-Guang Xie
中科院分区：
文献类型：
--
作者：
Ting Zhu;Yu Wu;Xue-Mei Li;Yu-Meng Jia;Huan Zhou;Li jiang;T. Tai;Qiong-Yu Mi;Jin-Zi Ji;Hong-Guang Xie
作为氯吡格雷和普拉格雷的类似物，在人体肠道中，主要通过AADAC和CES 2将维卡格雷完全水解为中间体硫内酯代谢物2-氧代-氯吡格雷（也是活性巯基代谢物H4的前体）;然而，其他未知的维卡格雷水解酶仍有待鉴定。本研究以重组人Raf激酶抑制蛋白（rhRKIP）、人肠道S9（HIS 9）组分和微粒体（HIM）制备物作为不同的酶源;普拉格雷作为RKIP的探针药物（阳性对照）、作为目标底物药物的维卡格雷以及作为水解酶活性指标的硫内酯代谢物2-氧代-氯吡格雷和R95913的形成速率，分别此外，还测定了洛可他汀抑制rhRKIP催化的维卡格雷水解的IC 50值，并使用了5种具有不同酯酶选择性的经典酯酶抑制剂来分析多种水解酶参与维卡格雷水解的情况。结果表明，rhRKIP在体外水解vicagrel，Km、Vmax和克林特值分别为20.04 ± 1.99 μM、434.60 ± 12.46 nM/min/mg蛋白和21.69 ± 0.28 ml/min/mg蛋白，而locostatin对rhRKIP的IC 50值估计为1.24 ± 0.04 mM。除了洛可他汀，艾司氯胺酮和长春碱强烈抑制在HIM中的维卡格雷水解。结果表明，RKIP可以催化人体肠道中的维格瑞洛水解，并且维格瑞洛可以被多种水解酶如RKIP、AADAC和CES 2同时水解。
As an analog of clopidogrel and prasugrel, vicagrel is completely hydrolyzed to intermediate thiolactone metabolite 2‐oxo‐clopidogrel (also the precursor of active thiol metabolite H4) in human intestine, predominantly by AADAC and CES2; however, other unknown vicagrel hydrolases remain to be identified. In this study, recombinant human Raf kinase inhibitor protein (rhRKIP) and pooled human intestinal S9 (HIS9) fractions and microsome (HIM) preparations were used as the different enzyme sources; prasugrel as a probe drug for RKIP (a positive control), vicagrel as a substrate drug of interest, and the rate of the formation of thiolactone metabolites 2‐oxo‐clopidogrel and R95913 as metrics of hydrolase activity examined, respectively. In addition, an IC50 value of inhibition of rhRKIP‐catalyzed vicagrel hydrolysis by locostatin was measured, and five classical esterase inhibitors with distinct esterase selectivity were used to dissect the involvement of multiple hydrolases in vicagrel hydrolysis. The results showed that rhRKIP hydrolyzed vicagrel in vitro, with the values of Km, Vmax, and CLint measured as 20.04 ± 1.99 μM, 434.60 ± 12.46 nM/min/mg protein, and 21.69 ± 0.28 ml/min/mg protein, respectively, and that an IC50 value of locostatin was estimated as 1.24 ± 0.04 mM for rhRKIP. In addition to locostatin, eserine and vinblastine strongly suppressed vicagrel hydrolysis in HIM. It is concluded that RKIP can catalyze the hydrolysis of vicagrel in the human intestine, and that vicagrel can be hydrolyzed by multiple hydrolases, such as RKIP, AADAC, and CES2, concomitantly.