Identifying SARS-CoV-2 antiviral compounds by screening for small molecule inhibitors of nsp13 helicase

Biochem J. 2021 Jul 16;478(13):2405-2423. doi: 10.1042/BCJ20210201.

Abstract

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.

Keywords: COVID-19; RNA helicase; coronavirus; nsp13.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antiviral Agents / chemistry*
  • Antiviral Agents / pharmacology*
  • Chlorocebus aethiops
  • Drug Evaluation, Preclinical*
  • Enzyme Assays
  • Fluorescence Resonance Energy Transfer
  • High-Throughput Screening Assays
  • RNA Helicases / antagonists & inhibitors*
  • RNA Helicases / metabolism
  • Reproducibility of Results
  • SARS-CoV-2 / drug effects
  • SARS-CoV-2 / enzymology*
  • Small Molecule Libraries / chemistry
  • Small Molecule Libraries / pharmacology*
  • Suramin / pharmacology
  • Vero Cells
  • Viral Nonstructural Proteins / antagonists & inhibitors*
  • Viral Nonstructural Proteins / metabolism

Substances

  • Antiviral Agents
  • Small Molecule Libraries
  • Viral Nonstructural Proteins
  • Suramin
  • RNA Helicases