α-葡萄糖苷酶抑制剂微量筛选模型的正交法构建和筛选

Study on microassay for screening α-glucosidase inhibitors by orthogonal matrix method

目的为新药发现阶段α-葡萄糖苷酶抑制剂的筛选提供实用的微量筛选方法。方法以α-葡萄糖苷酶为酶源,96孔板为载体,在生理pH值和温度的条件下,用正交试验法研究酶浓度、底物浓度、反应时间对酶活力的影响,确定酶反应最佳条件。在确定筛选模型运行条件后,对阳性药物和492种贵州民族药植物提取物进行筛选研究。结果采用正交试验法成功构建了实用灵敏的α-葡萄糖苷酶抑制剂微量筛选模型。492种植物提取物筛选结果表明,有145种提取物的抑制率达到了50%以上。结论本研究构建的α-葡萄糖苷酶抑制剂微量筛选模型具备了简便、快速、灵敏、高效、经济的优点,适用于大规模筛选α-葡萄糖苷酶抑制剂。

Aim To provide practical microassay for screeningα--glucosidase inhibitors in drug discovery. Methods The optimal conditions of assaying the activity of α--glucosidase were determined in 96-well plates under physiological pH value and temperature by orthogonal matrix method. Reaction time and the concentrations of α-glucosidase and substrate were optimized. After the effects of sample solvent （DMSO） used in the assay and stopping reagent on enzyme activity were assessed, the assay conditions were finally selected. Then 492 kinds of extracts from Guizhou ethno-drugs were screened. Results Practical and sensitive microassay for screening α-glucosidase inhibitors was successfully constituted. And in 492 kinds of extracts, 145 kinds of samples effectively inhibited the enzyme activity. Conclusion The microassay constituted in this work possesses advantages of being rapid, sensitive, rehable, cost saving, easy and flexible.