采用体外肝微粒体孵育体系,研究噻吩诺啡在大鼠、比格犬和人肝微粒体中酶代谢动力学及代谢产物差异。通过对噻吩诺啡浓度、微粒体蛋白含量和孵育时间等条件的考察优化噻吩诺啡与肝微粒体的反应体系;应用LC-MS/MS定量检测孵育体系中的噻吩诺啡及代谢产物,分析比较噻吩诺啡在3种肝微粒体中代谢产物种类和生成量的差异,计算并比较相应的动力学参数。噻吩诺啡在人肝微粒体中代谢转化最慢,其相应的动力学参数Km=(4.00±0.59)μmol·L-1、Vmax=(0.21±0.06)μmol·L-1·min-1、T1/2=(223±6.10)min、CLint=(117±3.19)mL·min-1·kg-1;比格犬和大鼠肝微粒体中相应的参数Km、Vmax、T1/2和CLint分别为(3.57±0.69)和(3.28±0.50)μmol·L-1、(0.18±0.04)和(0.14±0.04)μmol·L-1·min-1、(244±1.21)和(70.7±1.05)min、(213±1.06)和(527±7.79)mL·min-1·kg-1。在3个种属肝微粒体中均观察到噻吩诺啡的6个I相代谢产物,但6个产物的相对生成百分比在不同种属肝微粒体中有一定差异。实验结果表明,噻吩诺啡在体外人、比格犬和大鼠肝微粒体中主要的I相代谢途径相同,但是代谢产物的生成量及噻吩诺啡的代谢动力学性质存在着一定的差异。 The in vitro hepatic microsomal incubation system was used to study the differences in enzyme kinetics and metabolites of thienorphine in rat, beagle dog and human liver microsomes. The reaction system of thienorphine and liver microsome was optimized by investigating the conditions of thienorphine concentration, microsomal protein content and incubation time. The thienorphine and metabolites in the incubation system were determined by LC-MS / MS. Comparing the difference of the metabolites of thienorphine in the three kinds of liver microsomes and the amount of their metabolites, the corresponding kinetic parameters were calculated and compared. Thienorphine showed the slowest metabolic conversion in human liver microsomes. The corresponding kinetic parameters were Km = (4.00 ± 0.59) μmol·L-1, Vmax = (0.21 ± 0.06) μmol·L-1 · min- T1 / 2 = (223 ± 6.10) min and CLint = (117 ± 3.19) mL · min-1 · kg-1 respectively. The corresponding parameters Km, Vmax, T1 / 2 and CLint in beagle dogs and rat liver microsomes (3.57 ± 0.69) and (3.28 ± 0.50) μmol·L -1, (0.18 ± 0.04) and (0.14 ± 0.04) μmol·L -1 min -1, (244 ± 1.21) and (70.7 ± 1.05) ) min, (213 ± 1.06) and (527 ± 7.79) mL · min-1 · kg-1, respectively. Six I-phase metabolites of thienorphine were observed in the three species of liver microsomes, but the relative percentages of the six products were different among different species of liver microsomes. The experimental results show that the major phase I metabolic pathway of thienorphine in human liver, beagle dog and rat liver microsomes is the same, but there are some differences between the metabolites and the metabolites of thienorphine.