Identifying SARS-CoV-2 antiviral compounds by screening for small molecule inhibitors of nsp15 endoribonuclease.
Identifying SARS-CoV-2 antiviral compounds by screening for small molecule inhibitors of nsp15 endoribonuclease.
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DOI：
10.1042/bcj20210199
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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：
Canal B;Fujisawa R;McClure AW;Deegan TD;Wu M;Ulferts R;Weissmann F;Drury LS;Bertolin AP;Zeng J;Beale R;Howell M;Labib K;Diffley JFX
SARS-CoV-2是COVID-19的罪魁祸首，COVID-19是一种人类疾病，已造成200多万人死亡，使卫生系统接近崩溃，并危及世界各地国家和家庭的经济。目前缺乏对抗COVID-19的抗病毒治疗。Remdesivir是唯一被批准用于治疗COVID-19的抗病毒药物，它可以影响疾病的严重程度，但需要更好的治疗方法。SARS-CoV-2编码16个具有不同酶活性的非结构蛋白（non-structural proteins，nsp），这些蛋白在病毒基因组复制、转录和宿主免疫逃避中发挥重要作用。宿主免疫逃避的一个关键方面是通过nsp 15的尿苷指导的核糖核酸内切酶活性进行的。本研究对nsp 15重组蛋白的表达和纯化进行了研究。我们已经开发了生物化学测定来跟踪它的活性，我们已经发现了变构行为的证据。我们筛选了超过5000种化合物的定制化学文库以鉴定nsp 15内切核糖核酸酶抑制剂，并且我们鉴定并验证了NSC 95397作为nsp 15内切核糖核酸酶的体外抑制剂。虽然NSC 95397不能抑制SARS-CoV-2在VERO E6细胞中的生长，但需要进一步研究来确定nsp 15抑制对宿主免疫逃避的影响。
SARS-CoV-2 is responsible for COVID-19, a human disease that has caused over 2 million deaths, stretched health systems to near-breaking point and endangered economies of countries and families around the world. Antiviral treatments to combat COVID-19 are currently lacking. Remdesivir, the only antiviral drug approved for the treatment of COVID-19, can affect disease severity, but better treatments are needed. SARS-CoV-2 encodes 16 non-structural proteins (nsp) that possess different enzymatic activities with important roles in viral genome replication, transcription and host immune evasion. One key aspect of host immune evasion is performed by the uridine-directed endoribonuclease activity of nsp15. Here we describe the expression and purification of nsp15 recombinant protein. We have developed biochemical assays to follow its activity, and we have found evidence for allosteric behaviour. We screened a custom chemical library of over 5000 compounds to identify nsp15 endoribonuclease inhibitors, and we identified and validated NSC95397 as an inhibitor of nsp15 endoribonuclease in vitro. Although NSC95397 did not inhibit SARS-CoV-2 growth in VERO E6 cells, further studies will be required to determine the effect of nsp15 inhibition on host immune evasion.