Identifying SARS-CoV-2 antiviral compounds by screening for small molecule inhibitors of nsp13 helicase.
journal contributionposted on 2021-07-09, 09:41 authored by Jingkun Zeng, Florian Weissmann, Agustina P Bertolin, Viktor Posse, Berta Canal, Rachel Ulferts, Mary Wu, Ruth Harvey, Saira Hussain, Jennifer C Milligan, Chloe Roustan, Annabel Borg, Laura McCoy, Lucy S Drury, Svend Kjaer, John McCauley, Michael Howell, Rupert Beale, John FX Diffley
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.
Crick (Grant ID: 10827, Grant title: Beale FC001827) Crick (Grant ID: 10066, Grant title: Diffley FC001066) Crick (Grant ID: 10030, Grant title: McCauley FC001030) Crick (Grant ID: 10008, Grant title: STP High Throughput Screening) Crick (Grant ID: 10015, Grant title: STP Structural Biology) Wellcome Trust (Grant ID: 106252/Z/14/Z, Grant title: WT 106252/Z/14/Z) European Commission (Grant ID: 844211 - HUMAN REPL MECH, Grant title: EC 844211 - HUMAN REPL MECH)
coronavirusCOVID-19nsp13RNA helicaseAnimalsAntiviral AgentsChlorocebus aethiopsDrug Evaluation, PreclinicalEnzyme AssaysFluorescence Resonance Energy TransferHigh-Throughput Screening AssaysRNA HelicasesReproducibility of ResultsSARS-CoV-2Small Molecule LibrariesSuraminVero CellsViral Nonstructural ProteinsDiffley FC001066Beale FC001827McCauley FC001030HTSSBWIC03 Chemical Sciences06 Biological Sciences11 Medical and Health SciencesBiochemistry & Molecular Biology