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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。
Anti-hepatitis B candidate new drug Tiftamate (Y101) is a class of phenylalanine dipeptide derivatives, a new anti-hepatitis B virus mechanism and a good anti-hepatitis B virus. In this paper, a high-throughput cytochrome P450 (CYP) inhibition screening kit based on fluorescence assay was used to calculate the remaining enzyme activity and half-inhibitory concentration (IC50) by measuring the fluorescence intensity obtained, The results showed that Y101 had less inhibitory effect on CYP1A2, CYP3A4, CYP2C9, CYP2C19 and CYP2D6 (IC 50 was more than 100 μmol·L -1). Human primary cultured hepatocytes were cultured in Y101 for 72 h. The probe substrates of CYP1A2, CYP2B6 and CYP3A4 were mixed with “Cocktail” method and reacted by liquid chromatography-mass spectrometry (LC-MS / MS) At the same time, the concentrations of metabolites metabolized by the CYP isozymes of the probe substrate were measured to calculate the residual enzyme activity of CYP isoenzymes. Compared with the blank control group, the enzyme activity ratios of the experimental group and the blank control group were all less than 1 (between 0.662 and 0.928), which was less than 40% of the induction capacity compared with the positive control group. The results showed that Y101 had no effect on CYP1A2, CYP2B6 and CYP3A4 Potential. The results indicate that Y101 may not undergo metabolic interactions based on CYP enzyme inhibition or induction during combination therapy.