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1-(2-萘甲基)靛红-5-甲酰胺类化合物通过与底物口袋结合来抑制SARS-3CL蛋白酶的活性,而SARS-3CL蛋白酶自身的N端8肽是作用于蛋白二聚界面的抑制剂.本文设计同时占据SARS-3CL蛋白酶底物口袋和二聚界面的双功能抑制剂,通过固相多肽合成方法制备由1-(2-萘甲基)靛红-5-甲酸和N端8肽组成的化合物,得到不同长度连接链的6个目标产物.用显色底物方法测定化合物对SARS-3CL蛋白酶的抑制活性,其中化合物3的活性最高,IC50值(半抑制率)为3.8μmol·L-1,连接偶数甘氨酸的活性明显要好于连接奇数甘氨酸的化合物.用超速离心沉降速率方法研究了化合物3对SARS-3CL蛋白酶聚集状态与活性的调控作用,其同时具有诱导与抑制二聚的双重能力,综合调控结果是抑制SARS-3CL蛋白酶的二聚.这项研究给应用合成的化合物研究酶活性调节机制提供了一个示例.
The 1- (2-naphthylmethyl) isatin-5-carboxamides inhibit the activity of the SARS-3CL protease by binding to the substrate pocket, whereas the N-terminal 8-peptide of SARS-3CL protease acts on the dimerization of the protein Interfacial inhibitor.In this paper, we design a bifunctional inhibitor that simultaneously occupies both the pocket and the dimeric interface of the SARS-3CL protease substrate and prepares a novel peptide consisting of 1- (2-naphthylmethyl) isatin-5-carboxylic acid and N-terminal 8peptide to obtain six target products with different length of linker.The inhibitory activity of the compound on SARS-3CL protease was determined by colorimetric substrate method, among which compound 3 showed the highest activity and IC50 (half-inhibitory rate) Was 3.8μmol·L-1, and the activity of binding even glycine was significantly better than that of the compound with the excess of glycine.The compound 3 was used to control the aggregation status and activity of SARS-3CL protease by ultracentrifugal sedimentation rate method, Inhibition of the dual ability of dimerization and the combined result of regulation is to inhibit the dimerization of SARS-3CL protease.This study provides an example of the mechanism by which synthetic compounds can be used to study the activity of enzymes.