Identifying SARS-CoV-2 Antiviral Compounds by Screening for Small Molecule Inhibitors of Nsp12/7/8 RNA-dependent RNA Polymerase
| Autor: | Florian Weissmann, Berta Canal, Rachel Ulferts, Mary Wu, Michael Howell, John F.X. Diffley, Lucy S. Drury, Jennifer C. Milligan, Rupert Beale, Jingkun Zeng, Agustina P Bertolin, Viktor Posse |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
viruses Drug Evaluation Preclinical coronavirus Druggability Viral Nonstructural Proteins medicine.disease_cause RNA-dependent RNA polymerase Biochemistry Benzoates Chemical library chemistry.chemical_compound 0302 clinical medicine RNA polymerase Chlorocebus aethiops Fluorescence Resonance Energy Transfer Research Articles Coronavirus chemistry.chemical_classification Coronavirus RNA-Dependent RNA Polymerase biology Small molecule Suramin Antiviral Agents Small Molecule Libraries 03 medical and health sciences Biochemical Techniques & Resources Virology medicine Animals Molecular Biology Vero Cells Enzyme Assays SARS-CoV-2 Reproducibility of Results COVID-19 RNA virus Cell Biology Bridged Bicyclo Compounds Heterocyclic biology.organism_classification High-Throughput Screening Assays nsp12 030104 developmental biology Enzyme Viral replication chemistry Holoenzymes 030217 neurology & neurosurgery |
| Zdroj: | Biochemical Journal |
| DOI: | 10.1101/2021.04.07.438807 |
| Popis: | SummaryThe 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 homologs 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 (FRET)-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 3 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. |
| Databáze: | OpenAIRE |
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