人肝微粒体CYP亚型在新型吲哚醌类化合物629代谢中的作用

目的研究人肝微粒体重组体系中不同CYP亚型在丝裂霉素C(MMC)的衍生物5-氮丙啶-3-羟甲基-1-甲基吲哚-4,7-二酮[5-(aziridin-1-yl)-3-hydroxymethyl-1-methylin-dole-4,7-dione,简称629]代谢中的作用。方法不同浓度629与人肝脏微粒体共孵育,给予细胞不同CYP亚型特异性抑制剂的处理,用高压液相色谱法(high pressure liquid chro-matography,HPLC)分离、检测629的消失情况。结果HPLC检测到629在肝脏微粒体中的代谢遵循酶动力学剂量效应关系,Km值为336μmol·L-1。P450酶系中CYP1A2、CYP2B6和CYP2A6被抑制后,可影响629的代谢(P<0.05),并且3者中CYP1A2的影响较大,但三者间差异无显著性(P>0.05);而CYP3A4、CYP2C19、CYP2C9、CYP2E1和CYP2D6对629的代谢无影响(P>0.05)。结论新型吲哚醌类生物还原物629可在肝脏代谢,其中CYP1A2、CYP2B6和CYP2A6可参与629的代谢,这为新型生物还原活性物设计、开发及临床上的合理用药具有重大的意义。

Role of CYP hypotype on the metabolism of 629 in human liver microsomes in vitro

Aim To study the role of CYP hypotype in the metabolism of 629[5-(aziridin-1-yl)-3-hydroxymethyl-1-methylindole-4,7-dione]-the derivate of mitomycin-C(MMC) in human liver microsomes system in vitro.Methods 629 was incubated with human liver microsomes,and treated by specific inhibitors of different CYP hypotypes,then the parent disappearance of 629 was isolated and detected by High Pressure Liquid Chromatography(HPLC). Results 629 could be metabolized stably in human liver microsomes,It metabolized according to enzyme kinetics,and Km price was 336 μmol·L-1. CYP1A2,2B6 and CYP2A6 were inhibited,the metabolism of 629 was affected(P<0.05),but there was no effect on the metabolism of 629 by CYP3A4,2C19,2C9,2E1 and CYP2D6 (P>0.05). and the effect of CYP1A2 on 629 was slightly higher than the other two,but there were no significant difference among them(P>0.05). Conclusions In human liver microsome system in vitro,CYP1A2,2B6 and CYP2A6 contribute to the metabolism of 629.It is very important for bioreduction drugs design and development,and provide the basic experimental and theoretical profiles for extensive application in clinic.