以MIF为靶标的抑制剂药物高通量筛选模型的建立和应用

doi:10.13560/j.cnki.biotech.bull.1985.2016.09.034

摘要/Abstract

摘要： 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59.07 μmol/L、44.12 μmol/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。

关键词: 巨噬细胞迁移抑制因子, 高通量筛选, 互变异构酶活性, 抑制剂

Abstract: 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 59.07 μmol/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.

Key words: macrophage migration inhibitory factor, high-throughput screening, tautomerase activity, inhibitors

张晶, 孙瑞秋, 唐延婷, 刘祥. 以MIF为靶标的抑制剂药物高通量筛选模型的建立和应用[J]. 生物技术通报, 2016, 32(9): 253-259.

ZHANG Jing^{1, 2}, SUN Rui-qiu², TANG Yan-ting², LIU Xiang². Establishment and Application of a High-throughput Drug Screening Assay Targeting Macrophage Migration Inhibitory Factor[J]. Biotechnology Bulletin, 2016, 32(9): 253-259.

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