药物抑制CYP2D6突变体代谢右美沙芬的比较研究

目的：基于CYP2D6代谢右美沙芬成为去甲右美沙芬的反应，以去甲右美沙芬浓度为测定指标，考察CYP2D6突变对抑制剂响应作用的变化，为个体化用药提供参考。方法：选择亚洲人群中代表性的4种表型分别为CYP2D6*1、CYP2D6*2、CYP2D6*10和CYP2D6*39，以及6种CYP2D6抑制剂，包括奎尼丁、普罗帕酮、阿米替林、利培酮、氟伏沙明和美托洛尔。通过优化底物浓度、孵育时间和孵育酶量等反应参数，确定了孵育反应体系。利用LC-MS/MS方法测定上述孵育体系中去甲右美沙芬的浓度，计算得到了6种药物对4种CYP2D6酶的IC₅₀值。结果：建立并验证了去甲右美沙芬浓度的测定方法。得到了酶抑制结果：奎尼丁和普罗帕酮对野生酶（CYP2D6*1）具有最强的抑制作用，IC₅₀值分别为0.030μmol·L^-1和0.33μmol·L^-1；阿米替林、利培酮和氟伏沙明表现为中等抑制，IC₅₀值在6.0~8.0μmol·L^-1范围内；美托洛尔是最弱的抑制剂，IC₅₀值大于39.0μmol·L^-1。同种药物对CYP2D6*1、CYP2D6*2和CYP2D6*39的抑制作用没有显著区别。每种药物对CYP2D6*10的IC₅₀值是野生酶IC₅₀值的2.5~6.7倍。结论：CYP2D6*10突变体减弱了药物对其代谢右美沙芬的抑制能力，对于临床用药具有指导意义。  

Objective: To investigate the change of inhibitory capacity of six inhibitors on four CYP2D6 enzymes using the concentration of dextrorphan in incubation system as a measurement, which was based on the reaction of metabolism of dextromethorphan to dextromethorphan by CYP2D6 in order to provide references for individualized medication. Methods: Four most representative CYP2D6 enzymes in Asian population, such as CYP2D6*1, CYP2D6*2, CYP2D6*10 and CYP2D6*39 were chosen. Six CYP2D6 inhibitors, for example, quinidine, propafenone, amitriptyline, risperidone, fluvoxamine and metoprolol, were selected for inhibitorscreening. The incubation reaction system was determined by optimizing reaction parameters such as substrate concentration, incubation time, and amount of incubation enzyme. The LC-MS/MS method was used to determine the concentration of dextrorphan in the above incubation system. Finally, the various IC₅₀ values were compared among four CYP2D6 enzymes. Results: A method for determining the concentration of dextrorphan was developed and validated. From drug-enzyme inhibitory study, quinidine and propafenone were the most potent inhibitors towards wild type (CYP2D6*1)with IC₅₀ values of 0.030 and 0.33 μmol·L^-1, respectively. Then, amitriptyline, risperidone and fluvoxamine showed intermediate inhibitory effect with IC₅₀ values of 6.0-8.0 μmol·L^-1, while metoprolol showed the weakest inhibitory effect with IC₅₀ value more than 39.0 μmol·L^-1. In addition, compared with IC 50 values towards CYP2D6*1 for each drug, IC₅₀ values towards CYP2D6*10 were 2.5-6.7 fold higher than that of wild type. Whereas that towards CYP2D6*1, CYP2D6*2 and CYP2D6*39 exhibited minor difference. Conclusion: CYP2D6*10 enzyme reduces the inhibitory effect of six drugs on metabolizing dextromethorphan, which is of guiding significance for clinical medication. 

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