Probing the SAM Binding Site of SARS-CoV-2 Nsp14 In Vitro Using SAM Competitive Inhibitors Guides Developing Selective Bisubstrate Inhibitors.
Probing the SAM Binding Site of SARS-CoV-2 Nsp14 In Vitro Using SAM Competitive Inhibitors Guides Developing Selective Bisubstrate Inhibitors.
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DOI：
10.1177/24725552211026261
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发表时间：
2021-10
期刊：
影响因子：
--
通讯作者：
Vedadi M
中科院分区：
文献类型：
--
作者：
Devkota K;Schapira M;Perveen S;Khalili Yazdi A;Li F;Chau I;Ghiabi P;Hajian T;Loppnau P;Bolotokova A;Satchell KJF;Wang K;Li D;Liu J;Smil D;Luo M;Jin J;Fish PV;Brown PJ;Vedadi M
COVID-19大流行显然已将全球医疗保健系统带到了一个临界点，沿着带来了毁灭性的社会经济后果。引起这种疾病的SARS-CoV-2病毒利用RNA加帽来逃避人类免疫系统。非结构蛋白（Nonstructural protein，nsp）14是SARS-CoV-2中的16种nsp之一，在冠状形成过程中催化病毒RNA的N7-鸟苷甲基化。为了发现nsp 14甲基转移酶（MTase）活性的小分子抑制剂，我们开发并采用放射性MTase测定来筛选161个内部合成的S-腺苷甲硫氨酸（SAM）竞争性MTase抑制剂和SAM类似物的库。在6个鉴定的筛选命中中，SS 148抑制nsp 14 MTase活性，IC 50值为70 ± 6 nM，并且对20种人蛋白赖氨酸MTase具有选择性，表明SAM结合位点存在显著差异。有趣的是，IC 50值为1.1 ± 0.2 µM的DS 0464显示出双底物竞争性抑制剂作用机制。DS 0464对33种RNA、DNA和蛋白质MTase中的28种也具有选择性。由这些化合物提供的结构-活性关系应该指导选择性双底物nsp 14抑制剂的优化，并可能提供一种针对COVID-19和可能的其他冠状病毒的新型抗病毒药物的途径。
The COVID-19 pandemic has clearly brought the healthcare systems worldwide to a breaking point, along with devastating socioeconomic consequences. The SARS-CoV-2 virus, which causes the disease, uses RNA capping to evade the human immune system. Nonstructural protein (nsp) 14 is one of the 16 nsps in SARS-CoV-2 and catalyzes the methylation of the viral RNA at N7-guanosine in the cap formation process. To discover small-molecule inhibitors of nsp14 methyltransferase (MTase) activity, we developed and employed a radiometric MTase assay to screen a library of 161 in-house synthesized S-adenosylmethionine (SAM) competitive MTase inhibitors and SAM analogs. Among six identified screening hits, SS148 inhibited nsp14 MTase activity with an IC50 value of 70 ± 6 nM and was selective against 20 human protein lysine MTases, indicating significant differences in SAM binding sites. Interestingly, DS0464 with an IC50 value of 1.1 ± 0.2 µM showed a bisubstrate competitive inhibitor mechanism of action. DS0464 was also selective against 28 out of 33 RNA, DNA, and protein MTases. The structure–activity relationship provided by these compounds should guide the optimization of selective bisubstrate nsp14 inhibitors and may provide a path toward a novel class of antivirals against COVID-19, and possibly other coronaviruses.
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