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Km值,Vmax值和内在清除率Clint值(Vmax/Km)。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。
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（Km value, Vmax 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 ICs0 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.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2014年第9期1016-1020,共5页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
广州市医药卫生科技项目(20132A011015)
关键词
CYP2C19
基因多态性
荧光检测技术
CYP2C19
gene polymorphism
fluorogenic assay
enzyme kinetics
high throughput drug screen
