Identifying SARS-CoV-2 antiviral compounds by screening for small molecule inhibitors of Nsp14 RNA cap methyltransferase.
Identifying SARS-CoV-2 antiviral compounds by screening for small molecule inhibitors of Nsp14 RNA cap methyltransferase.
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DOI：
10.1042/bcj20210219
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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：
Basu S;Mak T;Ulferts R;Wu M;Deegan T;Fujisawa R;Tan KW;Lim CT;Basier C;Canal B;Curran JF;Drury LS;McClure AW;Roberts EL;Weissmann F;Zeisner TU;Beale R;Cowling VH;Howell M;Labib K;Diffley JFX
COVID-19大流行已成为世纪最严峻的公共卫生挑战之一，SARS-CoV-2是冠状病毒科家族中第三个导致人类致命疾病的成员。目前只有一种抗病毒化合物瑞德西韦可用于治疗COVID-19。为了确定其他潜在的治疗方法，我们研究了SARS-CoV-2基因组中编码的酶蛋白。在这项研究中，我们专注于病毒RNA帽甲基转移酶，它在使病毒蛋白翻译和促进病毒逃离免疫系统中发挥关键作用。我们表达并纯化了鸟嘌呤-N7甲基转移酶nsp 14和nsp 16 2′-O-甲基转移酶及其活化辅因子nsp 10。我们进行了体外高通量筛选抑制剂的nsp 14使用定制的化合物库超过5000药物化合物，以前已在临床或基础研究的特点。我们确定了四种化合物作为nsp 14的潜在抑制剂，所有这些化合物都在基于细胞的SARS-CoV-2感染模型中显示出抗病毒能力。四种化合物中的三种也表现出与Remdesivir对病毒复制的协同作用。
The COVID-19 pandemic has presented itself as one of the most critical public health challenges of the century, with SARS-CoV-2 being the third member of the Coronaviridae family to cause a fatal disease in humans. There is currently only one antiviral compound, remdesivir, that can be used for the treatment of COVID-19. To identify additional potential therapeutics, we investigated the enzymatic proteins encoded in the SARS-CoV-2 genome. In this study, we focussed on the viral RNA cap methyltransferases, which play key roles in enabling viral protein translation and facilitating viral escape from the immune system. We expressed and purified both the guanine-N7 methyltransferase nsp14, and the nsp16 2′-O-methyltransferase with its activating cofactor, nsp10. We performed an in vitro high-throughput screen for inhibitors of nsp14 using a custom compound library of over 5000 pharmaceutical compounds that have previously been characterised in either clinical or basic research. We identified four compounds as potential inhibitors of nsp14, all of which also showed antiviral capacity in a cell-based model of SARS-CoV-2 infection. Three of the four compounds also exhibited synergistic effects on viral replication with remdesivir.