Identifying SARS-CoV-2 antiviral compounds by screening for small molecule inhibitors of Nsp3 papain-like protease.
Identifying SARS-CoV-2 antiviral compounds by screening for small molecule inhibitors of Nsp3 papain-like protease.
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DOI：
10.1042/bcj20210244
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发表时间：
2021-07-16
期刊：
影响因子：
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通讯作者：
Diffley JFX
中科院分区：
文献类型：
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作者：
Lim CT;Tan KW;Wu M;Ulferts R;Armstrong LA;Ozono E;Drury LS;Milligan JC;Zeisner TU;Zeng J;Weissmann F;Canal B;Bineva-Todd G;Howell M;O'Reilly N;Beale R;Kulathu Y;Labib K;Diffley JFX
COVID-19大流行已成为本世纪最大的威胁生命的疾病。虽然疫苗接种应该提供一个长期的解决方案，但这是针对病毒突变的持续威胁，使目前的疫苗效果不佳。因此，小分子抗病毒剂将是非常有用的，以补充疫苗接种计划。COVID-19的病原体是一种新型冠状病毒SARS-CoV-2，它编码至少九种酶活性，这些酶活性都具有药物靶向潜力。nsp 3蛋白中含有的木瓜蛋白酶样蛋白酶（PLpro）从多蛋白前体产生病毒非结构蛋白，并切割泛素和ISG蛋白缀合物。在这里，我们描述PLpro的表达和纯化。我们开发了用于筛选定制化合物文库的蛋白酶测定，从中我们鉴定出二氢丹参酮I和Ro 08-2750为在蛋白酶和异肽酶测定中抑制PLpro并且还在基于细胞培养的测定中抑制病毒复制的化合物。
The COVID-19 pandemic has emerged as the biggest life-threatening disease of this century. Whilst vaccination should provide a long-term solution, this is pitted against the constant threat of mutations in the virus rendering the current vaccines less effective. Consequently, small molecule antiviral agents would be extremely useful to complement the vaccination program. The causative agent of COVID-19 is a novel coronavirus, SARS-CoV-2, which encodes at least nine enzymatic activities that all have drug targeting potential. The papain-like protease (PLpro) contained in the nsp3 protein generates viral non-structural proteins from a polyprotein precursor, and cleaves ubiquitin and ISG protein conjugates. Here we describe the expression and purification of PLpro. We developed a protease assay that was used to screen a custom compound library from which we identified dihydrotanshinone I and Ro 08-2750 as compounds that inhibit PLpro in protease and isopeptidase assays and also inhibit viral replication in cell culture-based assays.