Identification of SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of the nsp14 RNA Cap Methyltransferase
Identification of SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of the nsp14 RNA Cap Methyltransferase
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DOI：
10.1101/2021.04.07.438810
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发表时间：
2021-04
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通讯作者：
Souradeep Basu;Tiffany Mak;Rachel Ulferts;Mary Y. Wu;T. Deegan;R. Fujisawa;K. Tan;Chew Theng Lim;Clovis Basier;Berta Canal;Joseph F. Curran;L. Drury;Allison W. McClure;Emma L. Roberts;Florian Weissmann;Theresa U. Zeisner;R. Beale;V. Cowling;M. Howell;K. Labib;J. Diffley
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作者：
Souradeep Basu;Tiffany Mak;Rachel Ulferts;Mary Y. Wu;T. Deegan;R. Fujisawa;K. Tan;Chew Theng Lim;Clovis Basier;Berta Canal;Joseph F. Curran;L. Drury;Allison W. McClure;Emma L. Roberts;Florian Weissmann;Theresa U. Zeisner;R. Beale;V. Cowling;M. Howell;K. Labib;J. Diffley
COVID-19大流行已成为本世纪最关键的公共卫生挑战之一，SARS-CoV-2是冠状病毒科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remdesivir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，我们重点研究了病毒RNA帽甲基转移酶，它在使病毒蛋白翻译和促进病毒从免疫系统逃逸中起关键作用。我们表达并纯化了鸟嘌呤- n7甲基转移酶nsp14和nsp16 2 ' - o -甲基转移酶及其激活辅因子nsp10。我们对nsp14抑制剂进行了体外高通量筛选，使用了一个定制的化合物文库，其中包含超过5000种先前在临床或基础研究中表征的药物化合物。我们确定了4种化合物作为nsp14的潜在抑制剂，所有这些化合物在基于SARS-CoV-2感染的细胞模型中也显示出抗病毒能力。4种化合物中有3种还显示出与瑞德西韦的病毒复制协同作用。
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 fatal disease in humans. There is currently only one antiviral compound, remdesivir, that can be used for the treatment of COVID-19. In order 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 a key role 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 5,000 pharmaceutical compounds that have previously been characterised in either clinical or basic research. We identified 4 compounds as potential inhibitors of nsp14, all of which also show antiviral capacity in a cell based model of SARS-CoV-2 infection. Three of the 4 compounds also exhibited synergistic effects on viral replication with remdesivir.