甾体5α-还原酶抑制剂的高效液相色谱法体外筛选模型

Screen steroid 5 alpha-reductase inhibitors in vitro by high performance liquid chromatography

报道了一种用高效液相色谱法(HPLC)筛选甾体5α-还原酶抑制剂的体外模型.将甾体5α-还原酶液、睾酮溶液、辅酶(NADPH)与5α-还原酶抑制剂(非那甾胺和爱普列特)在37℃下反应30 min,反应终止后,通过HPLC测定反应前后底物睾酮量的变化来分析酶的活性,进而由酶活性的降低来判断抑制剂对5α-还原酶的抑制作用.将实验数据按Lineweaver-Burk倒数作图,求得甾体类抑制非那甾胺抑制常数Ki=4.32±0.38μmol/L,爱普列特抑制常数Ki=1.14±0.23μmol/L.实验所得的数据和图形均说明了本实验所采用的高效液相色谱法作为一种甾体5α-还原酶抑制剂体外筛选模型是简便可行的.

An in vitro method to screen steroid 5 alpha-reductase inhibitors by high performance liquid chromatography was reported. Steroid 5 alpha-reductase, series concentration of testosterone and Finasteride （or Epristeride） were incubated at 37℃ for 30 rain. Then the samples were analyzed by HPLC with C18 column. The descending rate of the concentration of testosterone showed the activity of the enzymatic. And inhibition constant （Ki） of the steroid 5α-reductatse inhibitors could be calculated according to the Lineweaver-Burk plots. The Ki values of Finasteride and Epristeride were Ki=4.32±3.8 μmol/L and 1. 14±0.23 μmol/L, respectively. The results indicated that the method is rapid and efficient to screen steroid 5 alpha-reductase inhibitors in vitro.