Enzyme Kinetics of TM-2 in Rat, Beagle Dog and Human Liver Microsomes
MEN Lei1,2a,2b, ZHAO Yun-li1, TANG Xing1, YU Zhi-guo1*
1. School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China; 2a. College of Life Science; 2b. Key Laboratory of Biotechnology and Bioresources Utilization, Ministry of Education, Dalian Minzu University, Dalian 116600, China
Abstract��OBJECTIVE To study the enzymatic kinetics of TM-2 in rat, Beagle dog and human liver microsomes by LC-MS/MS. METHODS TM-2 was incubated with liver microsomal incubation system. LC-MS/MS method was established for quantitative analysis of TM-2 with cabazitaxel as internal standard. The enzyme kinetics parameters Vmax and Km was calculated by the GraphPad Prism 5.0 software. RESULTS A rapid and sensitive LC-MS/MS method was developed to study the enzyme kinetics of TM-2 in rat, Beagle dog and human liver microsome. The corresponding enzymatic kinetic parameters in rat, Beagle dog and human were as follows: Vmaxvalues were 16.3, 354.6 and 154.8 nmol��min-1��mg(protein)-1, respectively; Km values were 25.7, 313.8 and 89.4 ��mol��L-1, respectively; CLint values were 0.63, 1.13 and 1.73 mL��min-1��mg(protein)-1, respectively. CONCLUSION The result indicates that there are species differences in the activity of metabolic enzyme and the affinity of TM-2 to the metabolic enzyme. Enzyme kinetic parameters obtained of TM-2 provide important parameters for the further study.
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MEN Lei, ZHAO Yun-li, TANG Xing, YU Zhi-guo. Enzyme Kinetics of TM-2 in Rat, Beagle Dog and Human Liver Microsomes. Chinese Pharmaceutical Journal, 2018, 53(8): 620-626.
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2018, Vol. 53 
