以MIF为靶标的抑制剂药物高通量筛选模型的建立和应用

Establishment and Application of a High-throughput Drug Screening Assay Targeting Macrophage Migration Inhibitory Factor

旨在以巨噬细胞迁移抑制因子(MIF)为靶标,采用紫外-分光光度法建立高通量药物筛选体系。对目的基因进行分子克隆,利用大肠杆菌原核表达系统进行纯化得到高纯度的目的蛋白,利用紫外-分光光度法构建酶活体系,并优化体系条件,建立合适的高通量药物筛选模型,最终从384种小分子中筛选出潜在的酶抑制剂。筛选模型构建成功,并筛选出酶活抑制率较高的小分子2种,测得半数抑制浓度IC50分别为59.07μmol/L、44.12μmol/L。针对MIF蛋白,建立了较理想的高通量药物筛选模型,适用于MIF蛋白酶活抑制剂的筛选,有利于后期的药物研发。

The aim is to establish a high-throughput drug screening assay targeting macrophage migration inhibitory factor （ MIF ） by UV-spectrophotometry. The target gene was molecularly cloned, prokaryotic expression system of Escherichia coli was used to purify, and then the highly-purified target protein was acquired. UV-spectrophotometry was utilized to establish the enzymatic system, in which then the conditions were optimized, and the proper high-throughput drug screening assay was set up and screened new MIF inhibitors from 384 small molecules. As results, the screening assay was established successfully, and 2 small molecules with high inhibitory rate were identified, while median inhibitory concentration （IC50） was 59.07μmol/L and 44.12 μmol/L, respectively. In conclusion, targeting MIF protein, the ideal high-throughput drug screening assay was established, which is appropriate for screening MIF inhibitors and conducive to the future drug development.