CYP2C19基因多态性在中草药物代谢性质筛选中的初步研究

中国临床药理学与治疗学 ›› 2014, Vol. 19 ›› Issue (9): 1016-1020.

CYP2C19基因多态性在中草药物代谢性质筛选中的初步研究

陈旺¹, 温纯洁², 付利娟²

¹ 暨南大学医学院第四附属医院、广州市红十字会医院药学部,广州 510220,广东;
² 重庆医科大学生命科学研究院,重庆 400016,重庆

收稿日期:2014-06-18 修回日期:2014-08-20 出版日期:2014-09-26 发布日期:2014-09-26
作者简介:陈旺,男,硕士研究生,主要从事临床药理学和遗传药理学研究。Tel: 18312015596 E-mail: cogito304@sina.com
基金资助:
广州市医药卫生科技项目(20132A011015)

Preliminary study of the CYP2C19 genotype in the application of Chinese herbal medicine metabolism screening

CHEN Wang¹, WEN Chun-jie², FU Li-juan²

¹ Department of Pharmacy, Guangzhou Red Cross Hospital, Medical College, Jinan University, Guangzhou 510220, China;
² Institute of Life Sciences, Chongqing Medical University, Chongqing 400016, Chongqing, China

Received:2014-06-18 Revised:2014-08-20 Online:2014-09-26 Published:2014-09-26

摘要/Abstract

摘要： 目的体外测定三个CYP2C19基因型(CYP2C19^*1、CYP2C19^*2、CYP2C19^*3)酶动力学常数和药物抑制常数,研究CYP2C19基因多态性对药物代谢的影响。方法以CEC(3-cynao-7-ethoxycoumarin)作为CYP2C19特异性荧光底物进行酶学反应,检测CYP2C19三个多态性酶的活性,使用非线性回归方法计算酶促反应动力学参数K_m值,V_max值和内在清除率Clint值(V_max/K_m)。通过将特异性荧光底物CEC,待检测中草药物以及其他用来模拟体内环境的相关实验条件相组合进行体外酶抑制反应,分析待检测的19种中草药物对CYP2C19基因多态性的抑制效应,并计算其IC₅₀值。结果抑制筛选实验表明:2C19参与代谢的底物和已知抑制剂均对其有抑制作用,多数中草药物对CYP2C19抑制作用微弱或者无抑制作用;化学结构或治疗作用类似的同类药品对CYP2C19的抑制情况不一;不同基因型被同一种药物抑制程度也不尽相同,甚至差别较大。结论建立了高效、稳定、简便的荧光高通量药物筛选平台,为后期研究CYP2C19应用于创新药物或新药先导化合物临床前代谢性质的筛选和预测奠定了基础,为临床潜在药物相互作用提供前期实验依据。

关键词: CYP2C19, 基因多态性, 荧光检测技术, 酶动力学分析, 药物高通量筛选

Abstract: AIM: To determine the constants of enzyme kinetics and drug inhibition of the three CYP2C19 genotype(CYP2C19^*1, CYP2C19^*2, CYP2C19^*3),and study the impact of CYP2C19 genetic polymorphism in drug metabolism in vitro. METHODS: In the study the CEC (3-cynao-7-ethoxycoumarin) as a CYP2C19-specific fluorescent substrate used for enzymatic reaction and detected CYP2C19 genotype activity, in addition, the nonlinear regression method to calculate the enzymatic kinetic parameters(K_m value, V_max of the value and intrinsicclear rate Clint value).Then, enzyme inhibition reaction analysis the inhibitory effect of 19 kinds of Chinese herbal medicine on the CYP2C19 genotypes, which by combined of fluorescent substrate CEC and herbal material, as well as other experimental conditions in vitro, and calculate the IC₅₀ values.RESULTS: The screening experiment results showed that: CYP2C19 involved in the metabolism of substrates and known inhibitors all were its inhibitor, majority of Chinese medicine monomer weak or no inhibitory effect on CYP2C19; chemical structure or similar drugs with similar therapeutic effects on inhibition of CYP2C19 different;the same drug inhibitied different genotypes were not the same result,evern a larger difference.CONCLUSION: Establish an efficient, stable, and easy fluorescent high-throughput drug screening platform, it bulited a basement applied before the metabolic nature of innovative drugs or new drug lead compounds for clinical screening and predictive for late-stage study,providing experimental basis for the clinical potential drug interactions.

Key words: CYP2C19, gene polymorphism, fluorogenic assay, enzyme kinetics, high throughput drug screen

中图分类号:

R969.1

陈旺, 温纯洁, 付利娟. CYP2C19基因多态性在中草药物代谢性质筛选中的初步研究[J]. 中国临床药理学与治疗学, 2014, 19(9): 1016-1020.

CHEN Wang, WEN Chun-jie, FU Li-juan. Preliminary study of the CYP2C19 genotype in the application of Chinese herbal medicine metabolism screening[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2014, 19(9): 1016-1020.

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