抗乙肝候选新药替芬泰对细胞色素P450酶的体外抑制和诱导作用研究  

The in vitro inhibition and induction of cytochrome P450 activities by bentysrepinine: a novel candidate of anti-hepatitis B virus drug

作　　者：樊慧蓉 慈小燕[2] 李薇[2] 董世奇[3] 曾勇[2] 伊秀林[2] 司端运[2] 刘昌孝[2] 

机构地区：[1]中国医学科学院、北京协和医学院放射医学研究所,天津市放射医学与分子核医学重点实验室,天津300192 [2]天津药物研究院,释药技术与药代动力学国家重点实验室,天津300193 [3]天津大学,天津300191

出　　处：《药学学报》2016年第12期1864-1870,共7页Acta Pharmaceutica Sinica

基　　金：科技部国家科技重大专项课题资助项目(2011ZX09102-009-02);国家自然科学基金重点项目(81430096;81302870);国家科技重大专项资助项目(2012ZX09506001-006);贵州省中药现代化科技产业研究开发专项项目([2010]5018)

摘　　要：抗乙肝候选新药替芬泰(Y101)是一类苯丙氨酸二肽衍生物,有全新的抗乙肝病毒机制和良好的抗乙肝病毒作用。本文应用基于荧光测定的高通量细胞色素P450酶(cytochrome P450,CYP)抑制筛选试剂盒,通过测定所得的荧光强度值,计算剩余酶活性及半数抑制浓度(IC50),评估Y101对CYP同工酶的抑制潜能,结果表明Y101对CYP1A2、CYP3A4、CYP2C9、CYP2C19和CYP2D6产生抑制作用的可能性较小(IC_(50)均大于100μmol·L^(-1))。人原代培养肝细胞中加入Y101培养72 h,以"Cocktail"法混合加入CYP1A2、CYP2B6和CYP3A4的探针底物进行反应,液相色谱-质谱联用(LC-MS/MS)法同时测定上述探针底物经CYP同工酶代谢的代谢产物浓度,计算CYP同工酶的剩余酶活性。实验组与空白对照组相比的酶活性比值均小于1(在0.662~0.928之间),与阳性对照组相比均小于其诱导能力的40%,结果表明Y101对CYP1A2、CYP2B6和CYP3A4无诱导潜能。结果表明,Y101可能不会在联合用药的过程中发生基于CYP酶抑制或诱导的代谢性相互作用。Bentysrepinine（Y101）, a derivative of phenyalanine dipeptide, has a novel mechanism in the treatment of hepatitis B virus（HBV） infection with a good anti-HBV effect. In the present study, a fluorometric-based high throughput method using cytochrome P450（CYP） screening kit was adopted to evaluate in vitro inhibition potential of Y101 on CYP isoenzymes by calculating remaining enzyme activities and inhibitory potential（IC50 values） using the determined values of fluorescence intensity. The result showed that Y101 exhibited little activity in the inhibition of CYP1A2, CYP3A4, CYP2C9, CYP2C19 and CYP2D6（IC50 100 μmol·L^-1）. Y101 was used to treat human primary hepotocytes for 72 h, and the enzyme activities of CYP1A2, CYP2B6 and CYP3A4 were determined with a cocktail of probe substrates for the three CYP isoforms. The metabolites were simultaneously determined using a LC-MS/MS method. Y101 had no activity in the induction of CYP1A2, CYP2B6 and CYP3A4 on the basis of the following results： 1 The ratio of enzyme activities between test and control groups were all below than 1（varied from 0.662 to 0.928）; 2 The induction potential of Y101 were lower than forty percent compared with that of positive groups. The above results suggest that Y101 has little activity in the regulation of metabolic drug-drug interactions based on the CYP isoform changes following co-administration of drugs.

关 键 词：替芬泰 CYP450同工酶 酶活性 抑制 诱导 药物-药物相互作用 

分 类 号：R963[医药卫生—微生物与生化药学]