UPLC-MS/MS研究抗肿瘤化合物HK-7在不同种属肝微粒体中的代谢稳定性和代谢酶表型

Investigation of metabolic stability and metabolic enzyme phaenotypes of anti-tumor compound HK-7 in different species of liver microsomes by UPLC-MS/MS

目的:应用体外肝微粒体孵育体系,研究抗肿瘤化合物HK-7在人、SD大鼠、昆明种小鼠、恒河猴和比格犬的肝微粒体中的代谢稳定性,确定HK-7在人肝微粒体中的代谢表型。方法:通过UPLC-MS/MS检测方法,测定在各个种属中孵育后的HK-7剩余浓度,考察它们的代谢稳定性及体外代谢动力学参数。采用化学抑制剂法预测HK-7在人肝微粒体中的代谢表型。结果:在人、SD大鼠、昆明种小鼠、恒河猴和比格犬的肝微粒体中,HK-7的体外代谢半衰期t1/2分别为44.91,83.42,72.70,48.93和66.21 min;体外固有清除率CLint分别为0.031,0.016,0.019,0.028和0.021 m L·min^(-1)·mg^(-1);在人肝微粒体中HK-7主要被细胞色素CYP2C9,CYP2E1和CYP3A4催化代谢。结论:HK-7在肝微粒体中的代谢存在种属差异,人肝微粒体中参与HK-7代谢的酶可能有CYP2C9,CYP2E1和CYP3A4,其中CYP2E1贡献最大。

Objective： To investigate the metabolic stability of HK-7 in liver microsomes of human, SD rats, Kunming mice, Rhesus monkeys and Beagle dogs, and identify the main enzymes involved in HK-7 metabolism in human liver microsomes. Methods： After being incubated with different species of liver microsomes, HK-7 was quantified by UPLC-MS/MS method to evaluate its stability and metabolic kinetic parameters in vitro. The CYP450 phenotype of HK-7 was identified by specific inhibitors of isoforms in human microsomal incubation system. Results： The in vitro half-lives （t1/2 ） of HK-7 in liver microsomes of human, SD rats, Kunming mice, Rhesus monkeys and Beagle dogs were 44.91,83.42,72.70,48.93, and 66.21 rain respectively. Their intrinsic clearance rates （CLint） were0. 031, 0.016, 0.019, 0.028, and0.021 mL-min l.mg-1 respectively. CYP2C9 CYP2E1, and CYP3A4 were found to be the major CYP isoforms involved in HK-7 metabolism. Conclusion： There is significant difference in the metabolic rate of HK-7 between different species. The metabolism of HK-7 in human liver microsomes may be CYP2C9, CYP2E1 and CYP3A4, and CYP2E1 has the greatest contribution to its metabolism.