In Vitro Assessment of 36 CYP2C9 Allelic Isoforms Found in the Chinese Population on the Metabolism of Glimepiride
In Vitro Assessment of 36 CYP2C9 Allelic Isoforms Found in the Chinese Population on the Metabolism of Glimepiride
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中国人群中发现的 36 个 CYP2C9 等位基因亚型对格列美脲代谢的体外评估
DOI：
10.1111/bcpt.12159
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发表时间：
2014-04-01
影响因子：
3.1
通讯作者：
Cai, Jian-Ping
中科院分区：
文献类型：
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作者：
Dai, Da-Peng;Wang, Shuang-Hu;Cai, Jian-Ping
到目前为止，在已报道的57个CYP2C9等位基因中，有36个已在中国人群中被鉴定出来。本研究的目的是评估这些等位基因异构体的催化特性及其对格列美脲体外代谢的影响。利用杆状病毒介导的表达系统在昆虫细胞微粒体中高效表达野生型及其35个等位基因突变体。然后，以格列美脲为底物，对各变异体的酶学性质进行了评价。与昆虫微粒子和0.125-10μM格列美脲在37℃下反应40min。终止后，提取产物，用LC-MS/MS进行信号采集。在所检测的36个等位基因异构体中，只有4个变异株(CYP2C9.40、CYP2C9.47、CYP2C9.51和CYP2C9.54)的相对清除值与野生型的相近。此外，一个变异体(CYP2C9.36)的固有清除值高于野生型蛋白，而其余30个等位基因亚型的清除值明显低于野生型蛋白(从0.1%到87.2%)。这项研究提供了关于中国人群中关于常用降糖药物格列美脲的所有已报告的CYP2C9变异体的酶活性的最全面的数据。我们的结果表明，大多数被测试的稀有等位基因在体外显着降低了CYP2C9变体对格列美脲羟化的催化活性。
Of the 57 reported CYP2C9 alleles, to date, 36 of them have been identified in the Chinese population. The aim of this study was to assess the catalytic characteristics of these allelic isoforms and their effects on the metabolism of glimepiride in vitro. Baculovirus‐mediated expressing system was used to highly express wild‐type and the 35 CYP2C9 allelic variants in insect cell microsomes. Then, the enzymatic characteristics of each variant were evaluated using glimepiride as the substrate. Reactions were performed at 37°C with the insect microsomes and 0.125–10 μM glimepiride for 40 min. After termination, the products were extracted and used for signal collection by LC‐MS/MS. Of the 36 tested CYP2C9 allelic isoforms, only four variants (CYP2C9.40, CYP2C9.47, CYP2C9.51 and CYP2C9.54) exhibited similar relative clearance values to that of wild‐type CYP2C9.1. In addition, one variant (CYP2C9.36) showed a higher intrinsic clearance value than the wild‐type protein, while the remaining 30 CYP2C9 allelic isoforms exhibited significantly decreased clearance values (from 0.1% to 87.2%) compared to CYP2C9.1. This study provided the most comprehensive data on the enzymatic activities of all reported CYP2C9 variants in the Chinese population with regard to the commonly used antidiabetic drug, glimepiride. Our results indicate that most of the tested rare alleles significantly decrease the catalytic activity of CYP2C9 variants towards glimepiride hydroxylation in vitro.