Chromosome Replication Laboratory, The Francis Crick Institute, 1 Midland Road, London NW1 1AT, U.K.
Cell Biology of Infection Laboratory, The Francis Crick Institute, 1 Midland Road, London NW1 1AT, U.K.
Biochem J. 2021 Jul 16;478(13):2405-2423. doi: 10.1042/BCJ20210201.
The coronavirus disease 2019 (COVID-19) pandemic, which is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is a global public health challenge. While the efficacy of vaccines against emerging and future virus variants remains unclear, there is a need for therapeutics. Repurposing existing drugs represents a promising and potentially rapid opportunity to find novel antivirals against SARS-CoV-2. The virus encodes at least nine enzymatic activities that are potential drug targets. Here, we have expressed, purified and developed enzymatic assays for SARS-CoV-2 nsp13 helicase, a viral replication protein that is essential for the coronavirus life cycle. We screened a custom chemical library of over 5000 previously characterized pharmaceuticals for nsp13 inhibitors using a fluorescence resonance energy transfer-based high-throughput screening approach. From this, we have identified FPA-124 and several suramin-related compounds as novel inhibitors of nsp13 helicase activity in vitro. We describe the efficacy of these drugs using assays we developed to monitor SARS-CoV-2 growth in Vero E6 cells.
新型冠状病毒病（COVID-19）大流行是由严重急性呼吸系统综合征冠状病毒 2（SARS-CoV-2）引起的，是一项全球性的公共卫生挑战。虽然针对新出现和未来病毒变异体的疫苗的功效尚不清楚，但仍需要治疗方法。重新利用现有药物是寻找针对 SARS-CoV-2 的新型抗病毒药物的一种很有前途且潜在快速的机会。该病毒编码至少 9 种酶活性，这些酶活性是潜在的药物靶点。在这里，我们已经表达、纯化并开发了 SARS-CoV-2 nsp13 解旋酶的酶促测定方法，nsp13 解旋酶是一种病毒复制蛋白，对冠状病毒的生命周期至关重要。我们使用基于荧光共振能量转移的高通量筛选方法，对超过 5000 种先前经过表征的药物的定制化学文库进行了 nsp13 抑制剂的筛选。由此，我们鉴定了 FPA-124 和几种苏拉明相关化合物作为 nsp13 解旋酶体外活性的新型抑制剂。我们使用我们开发的监测 Vero E6 细胞中 SARS-CoV-2 生长的测定方法来描述这些药物的功效。
