Remdesivir is a direct-acting antiviral that inhibits RNA-dependent RNA polymerase from severe acute respiratory syndrome coronavirus 2 with high potency
Remdesivir is a direct-acting antiviral that inhibits RNA-dependent RNA polymerase from severe acute respiratory syndrome coronavirus 2 with high potency
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DOI：
10.1074/jbc.ra120.013679
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发表时间：
2020-05-15
影响因子：
4.8
通讯作者：
Gotte, Matthias
中科院分区：
文献类型：
--
作者：
Gordon, Calvin J.;Tchesnokov, Egor P.;Gotte, Matthias
迫切需要对冠状病毒病2019年(新冠肺炎)进行有效治疗，以控制由严重急性呼吸综合征冠状病毒2号(SARS-CoV-2)引起的当前大流行。SARS-CoV-2的复制依赖于病毒RNA依赖的RNA聚合酶(RdRp)，而RdRp可能是研究核苷酸类似物雷米德韦尔(RDV)的靶标。RDV对RNA病毒表现出广谱的抗病毒活性，先前对来自埃博拉病毒和中东呼吸综合征冠状病毒(MERS-CoV)的RdRp的研究表明，延迟链终止是RDV可能的作用机制。在此，我们表达并纯化了由非结构蛋白NSP8和NSP12组成的活性SARS-CoV-2RdRp。酶动力学分析表明，该RdRp能有效地将RDV的活性三磷酸形式(RDV-TP)整合到RNA中。在I位掺入RDV-TP导致I+3位RNA合成终止。我们得到了与SARS-CoV、MERS-CoV和SARS-CoV-2 RdRps几乎相同的结果。RDV-TP的一个独特性质是它比天然核苷酸对应物ATP的掺入具有高的选择性。在这方面，被批准用于丙型肝炎病毒感染管理的2？-C-甲基化化合物的三磷酸形式，包括索莫布韦，以及广泛作用的抗病毒药物法韦拉韦和利巴韦林，显示出显著的缺陷。此外，我们为RDV的靶标特异性提供了证据，因为RDV-TP被远亲的Lassa病毒RdRp低效率地结合，并且没有观察到RNA合成的终止。这些结果共同为RDV介导的冠状病毒RNA合成抑制提供了一个统一的、精细的机制，并将这种核苷酸类似物定义为直接作用的抗病毒药物。
Effective treatments for coronavirus disease 2019 (COVID-19) are urgently needed to control this current pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Replication of SARS-CoV-2 depends on the viral RNA-dependent RNA polymerase (RdRp), which is the likely target of the investigational nucleotide analogue remdesivir (RDV). RDV shows broad-spectrum antiviral activity against RNA viruses, and previous studies with RdRps from Ebola virus and Middle East respiratory syndrome coronavirus (MERS-CoV) have revealed that delayed chain termination is RDV's plausible mechanism of action. Here, we expressed and purified active SARS-CoV-2 RdRp composed of the nonstructural proteins nsp8 and nsp12. Enzyme kinetics indicated that this RdRp efficiently incorporates the active triphosphate form of RDV (RDV-TP) into RNA. Incorporation of RDV-TP at position i caused termination of RNA synthesis at position i+3. We obtained almost identical results with SARS-CoV, MERS-CoV, and SARS-CoV-2 RdRps. A unique property of RDV-TP is its high selectivity over incorporation of its natural nucleotide counterpart ATP. In this regard, the triphosphate forms of 2?-C-methylated compounds, including sofosbuvir, approved for the management of hepatitis C virus infection, and the broad-acting antivirals favipiravir and ribavirin, exhibited significant deficits. Furthermore, we provide evidence for the target specificity of RDV, as RDV-TP was less efficiently incorporated by the distantly related Lassa virus RdRp, and termination of RNA synthesis was not observed. These results collectively provide a unifying, refined mechanism of RDV-mediated RNA synthesis inhibition in coronaviruses and define this nucleotide analogue as a direct-acting antiviral.