采用毛细管电泳方法以凝血酶为靶筛选天然药物提取化合物

Application of Capillary Electrophoresis to Screening Natural Products Using Thrombin as Target

采用毛细管电泳(CE)法研究了凝血酶和水蛭素及肝素间的相互作用,并测定了凝血酶和水蛭素的结合常数.选择凝血酶为靶点,建立了快速灵敏的从天然植物提取物中筛选出先导化合物的CE方法.在接近生理环境的水溶液条件下,研究了两色金鸡菊、肉苁蓉和黄蜀葵中天然植物提取物与凝血酶之间的相互作用;与阴性对照和阳性对照相比,从11个天然化合物中快速筛选出与凝血酶有相互作用的6个化合物,提高了天然植物提取物药理活性筛选的效率.

In this article, the interactions between hirudin and thrombin, and that between heparin and thrombin were investigated by using capillary electrophoresis. The binding constant between hirudin and thrombin was calculated to be 7.85 × 10^5 L/mol. A capillary zone electrophoretic method was applied to determine the interactions between natural products and thrombin. The samples containing the natural extractions and thrombin at various ratios were incubated at 25 ℃ and then were separated by CZE with Tris-acetate buffer at pH =7.2. Each run could be accomplished within 10 min. The experimental results show that the reported method by capillary zone electrophoresis for the determination of the interactions between natural products and thrombin is powerful, sensitive and fast, requires less amounts of reagents, and further, it can be employed as a reliable alternative to other methods.