UFLC-MS/MS研究胡椒碱在不同种属肝微粒体中的代谢稳定性和代谢表型

应用体外肝微粒体孵育体系,考察胡椒碱在人、SD大鼠、小鼠、恒河猴和比格犬5个种属肝微粒体中的代谢稳定性,比较代谢的种属差异,确定其在人肝微粒体中的代谢表型。通过UFLC-MS/MS检测方法,测定胡椒碱在各个种属肝微粒体中孵育后的剩余浓度,考察他们的代谢稳定性及体外代谢动力学参数。采用化学抑制法考察胡椒碱在人肝微粒体中的代谢表型。结果表明胡椒碱在人、SD大鼠、小鼠、恒河猴和比格犬的肝微粒体中,半衰期T1/2分别为31. 36、48. 46、138. 60、147. 45、165. 00 min;体外固有清除率CLint分别为0. 0442、0. 0286、0. 0100、0. 0094、0. 0084m L/(m L·mg);在人肝微粒体中,胡椒碱主要被CYP3A4和CYP2C9酶代谢。推测胡椒碱在各种肝微粒体中的代谢均相对较稳定,其中大鼠和人的肝微粒体代谢性质最相近,在后续的实验中可以考虑用大鼠的代谢结果预测人的代谢结果;人肝微粒体中参与胡椒碱代谢的酶主要有CYP3A4和CYP2C9。

Investigation of metabolic stability and metabolic enzyme phenotype of piperine in different species of liver microsomes by UFLC-MS/MS

In vitro liver microsomal incubation system was used to investigate the metabolic stability of piperine in liver microsomes of human,SD rats,mice,Rhesus monkeys and Beagle dogs,compare the metabolic difference between differenct species and determine the metabolic phenotype of piperine in human liver microsomes.Selective chemical inhibition was used to investigate the metabolic phenotype of piperine in human liver microsomes.The remaining concentrations of piperine incubated in various species of liver microsomes were determined by UFLC-MS/MS,and evaluated their metabolic stability and in vitro pharmacokinetic parameters.Selective chemical inhibition was used to predict the metabolic phenotype of piperine in human liver microsomes.It can be speculated that the half-life of piperine in the liver microsomes of SD rats,mice,rhesus monkeys,and Beagle dogs respectively were 31.36,48.46,138.60,147.45,and 165.00 min;The ininsic clearance rate was 0.044 2,0.028 6,0.010 0,0.009 4 and 0.008 4 mL/(mL·mg);In human liver microsomes,piperine was mainly metabolized by CYP3A4 and CYP2C9 enzymes.The results showed that the metabolism of piperine in all kinds of liver microsomes is relatively stable,the metabolic properties of liver microsomes in rat is similar mostly to human.It can be considered that metabolism results from rats can be used to predict human metabolic results in subsequent experiments.The results showed that CYP3A4 and CYP2C9 primary participate in metabolism of piperine.