Identifying SARS-CoV-2 antiviral compounds by screening for small molecule inhibitors of nsp12/7/8 RNA-dependent RNA polymerase.
Identifying SARS-CoV-2 antiviral compounds by screening for small molecule inhibitors of nsp12/7/8 RNA-dependent RNA polymerase.
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DOI：
10.1042/bcj20210200
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发表时间：
2021-07-16
期刊：
影响因子：
--
通讯作者：
Diffley JFX
中科院分区：
文献类型：
--
作者：
Bertolin AP;Weissmann F;Zeng J;Posse V;Milligan JC;Canal B;Ulferts R;Wu M;Drury LS;Howell M;Beale R;Diffley JFX
2019冠状病毒病（COVID-19）全球大流行已演变为近代史上最大的公共卫生和经济危机，几乎影响到社会的所有领域。需要有效的治疗方法来对抗正在进行的大流行。重新利用具有已知药理学安全性的现有药物是一种快速且具有成本效益的方法来确定新的治疗方法。COVID-19的病原体是严重急性呼吸综合征冠状病毒2（SARS-CoV-2），一种单链正义RNA病毒。冠状病毒依赖于复制-转录复合物（RTC）的酶活性在宿主细胞内繁殖。RTC核心催化组分是RNA依赖性RNA聚合酶（RdRp）全酶。由于RdRp在病毒复制中的重要作用、高度的序列和结构保守性以及在人类细胞中缺乏同源物，因此RdRp是CoV的关键药物靶标之一。在这里，我们表达，纯化和生化特性的活性SARS-CoV-2 RdRp复合物。我们开发了一种新的荧光共振能量转移为基础的链置换检测SARS-CoV-2 RdRp活性适合高通量格式。作为一个更大的研究项目的一部分，以确定抑制剂的所有酶活性编码的SARS-CoV-2，我们使用这种方法来筛选一个定制的化学库超过5000批准和研究化合物的新型SARS-CoV-2 RdRp抑制剂。我们鉴定了三个新化合物（GSK-650394、C646和BH 3 I-1），并证实苏拉明和苏拉明样化合物是体外SARS-CoV-2 RdRp活性抑制剂。我们还在我们开发的用于监测SARS-CoV-2生长的基于细胞的测定中表征了这些药物的抗病毒功效。
The coronavirus disease 2019 (COVID-19) global pandemic has turned into the largest public health and economic crisis in recent history impacting virtually all sectors of society. There is a need for effective therapeutics to battle the ongoing pandemic. Repurposing existing drugs with known pharmacological safety profiles is a fast and cost-effective approach to identify novel treatments. The COVID-19 etiologic agent is the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a single-stranded positive-sense RNA virus. Coronaviruses rely on the enzymatic activity of the replication–transcription complex (RTC) to multiply inside host cells. The RTC core catalytic component is the RNA-dependent RNA polymerase (RdRp) holoenzyme. The RdRp is one of the key druggable targets for CoVs due to its essential role in viral replication, high degree of sequence and structural conservation and the lack of homologues in human cells. Here, we have expressed, purified and biochemically characterised active SARS-CoV-2 RdRp complexes. We developed a novel fluorescence resonance energy transfer-based strand displacement assay for monitoring SARS-CoV-2 RdRp activity suitable for a high-throughput format. As part of a larger research project to identify inhibitors for all the enzymatic activities encoded by SARS-CoV-2, we used this assay to screen a custom chemical library of over 5000 approved and investigational compounds for novel SARS-CoV-2 RdRp inhibitors. We identified three novel compounds (GSK-650394, C646 and BH3I-1) and confirmed suramin and suramin-like compounds as in vitro SARS-CoV-2 RdRp activity inhibitors. We also characterised the antiviral efficacy of these drugs in cell-based assays that we developed to monitor SARS-CoV-2 growth.
影响因子：
8.8
作者：
Hosoi, Y;Matsumoto, Y;Tomita, M;Enomoto, A;Morita, A;Sakai, K;Umeda, N;Zhao, H-J;Nakagawa, K;Ono, T;Suzuki, N
通讯作者：
Suzuki, N