论文部分内容阅读
目的:以重组胶原酶催化区建立高通量药物筛选模型,筛选胶原酶抑制剂.方法:在大肠杆菌中表达了人的一型胶原酶的催化区,并以此建立了高通量药物筛选模型.对一批共2720个纯化合物进行了高通量筛选.结果:胶原酶催化区的高通量筛选中,完成2720个纯化合物的筛选只用了2小时10分,每个化合物的用量是4μg.高通量筛选发现66个化合物在20 mg/L的浓度下具有大于60％的抑制活性,其中有44个化合物的抑制活性被随后的多浓度测试所证实.筛选发现的最好化合物的IC_(50)为4.3μmol/L,共有IC_(50)小于20μmol/L的化合物15个.结论:重组胶原酶催化区的高通量药物筛选模型具有快速、微量、准确的特点.
OBJECTIVE: To establish a high-throughput drug screening model using collagenase catalytic domain and screen collagenase inhibitor.Methods: The human collagenase catalytic domain was expressed in Escherichia coli and established a high-throughput drug screening Model. A total of 2,720 pure compounds were subjected to high-throughput screening.Results: In a high-throughput screening of the collagenase catalytic zone, the screening of 2,720 pure compounds took only 2 hours and 10 minutes and the amount of each compound Was 4 The high throughput screening found that 66 compounds had greater than 60% inhibitory activity at a concentration of 20 mg / L, of which 44 compounds were tested for inhibitory activity as demonstrated by subsequent multi-concentration tests.Screening of the best compounds found (IC 50) of 4.3 μmol / L, and 15 compounds with IC 50 less than 20 μmol / L.Conclusion: The high-throughput drug screening model of the recombinant collagenase catalytic zone has the characteristics of rapid, trace and accurate.