【摘要】 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。更多还原

【Abstract】 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 IC₅₀values 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 0.33μmol·L^-1,respectively.Then,amitriptyline,risperidone and fluvoxamine showed intermediate inhibitory effect with IC₅₀ values of 6.0-8.0μmol·L^-1,while metoprolol showed the weakest inhibitory effect with IC₅₀ value more than 39.0μmol·L^-1.In addition,compared with IC₅₀values 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.更多还原