大鼠肝微粒体中有机锡抗癌化合物DBDCT的测定与酶促动力学研究

Determination of Antitumor Diorganotin Compound DBDCT in Rat Liver Microsomes by HPLC and Its Enzymatic Kinetics

关键词:  DBDCT  高效液相色谱法  大鼠肝微粒体  酶促动力学

Key words:  DBDCT  HPLC  rat liver microsomes  enzymatic kinetics

摘要:

目的 建立测定大鼠肝微粒体中有机锡抗癌化合物DBDCT的方法并研究其相应的酶促动力学。方法 采用HPLC测定DBDCT在体外代谢系统中的酶促动力学，用Eadie-Hofstee法分析数据，求算酶促动力学参数Km和Vmax以及肝代谢速率Clin。结果 在体外代谢系统中，DBDCT代谢酶促动力学参数Km=159.07 μmol·L^-1，Vmax=2.813 μmol·(min·mg)^-1以及肝清除率Clin=V_max/Km=0.017 L·(min·mg)^-1。结论 此分析方法符合方法学验证要求且简单准确，可满足DBDCT体外代谢的研究。

Abstract:

OBJECTIVE To establish an HPLC method for the determination of an antitumor diorganotin(IV) compound DBDCT and study its enzymatic kinetics in rat liver microsomes. METHODS HPLC was employed to determine the enzyme kinetics of DBDCT in vitro metabolic system using the rat liver microsomes. The kinetic parameters, Km, Vmax and liver clearance rate (Vmax/Km)were calculated by Eadie-Hofstee plot. RESULTS In the metabolic system, the enzyme kinetics parameters of DBDCT were found as follows: Km=157.07 μmol·L^-1, V_max=2.189 μmol·(min·mg)^-1 and liver clearance rate Vmax/Km=0.014 L·(min·mg)^-1, respectively. CONCLUSION The method is satisfied to the requirement of methodological verification, and it is simple, accurate and can be used for the in vitro metabolism research of DBDCT.