白介素-1β转化酶抑制剂的高通量筛选

High-throughput screening for interleukin-1β converting enzyme inhibitors

目的筛选具有白介素-1β转化酶(ICE)抑制活性的化合物。方法通过体外克隆ICE原酶全长基因、质粒原核表达及蛋白纯化等方式,应用酶与特异性底物的荧光反应,建立ICE抑制剂高通量筛选平台;通过紫外、质谱及核磁等理化数据分析对所筛选的化合物进行结构鉴定,同时进行了初步的药理毒理学活性验证。结果筛选得到1种具有较强ICE抑制活性的化合物LX1986,该化合物与已知化合物棕曲菌素D(Aspochalasin D)结构相同。结论 LX1986为活性较强的微生物来源ICE抑制剂,其抑制活性为首次报道。

Objective To screen compounds with interleukin-1β converting enzyme （ICE） inhibiting activity. Methods The high throughput screenirLg （HTS） assay for ICE inhibitor was established. It based on the clone of ICE pro-enzymes genes in vitro, prokaryotic expression, the protein purification and the fluorescence reaction between ICE and specific substrate. By the data of UV, MS and NMR, the structure of the selected compound was determined. Meanwhile, we did some pharmaco-toxicological evaluation in vitro. Results An active compound named LX1986 showed positive activity. The structure of the compound was the same with the Aspochalasin D. Conclusion LX1986 is a specific strong ICE inhibitor and the ICE inhibitor activity is the first reported.