Targeted design of drug binding sites in the main protease of SARS-CoV-2 reveals potential signatures of adaptation

Biochem Biophys Res Commun. 2021 May 28:555:147-153. doi: 10.1016/j.bbrc.2021.03.118. Epub 2021 Mar 26.


Several existing drugs are currently being tested worldwide to treat COVID-19 patients. Recent data indicate that SARS-CoV-2 is rapidly evolving into more transmissible variants. It is therefore highly possible that SARS-CoV-2 can accumulate adaptive mutations modulating drug susceptibility and hampering viral antigenicity. Thus, it is vital to predict potential non-synonymous mutation sites and predict the evolution of protein structural modifications leading to drug tolerance. As two FDA-approved anti-hepatitis C virus (HCV) drugs, boceprevir, and telaprevir, have been shown to effectively inhibit SARS-CoV-2 by targeting the main protease (Mpro), here we used a high-throughput interface-based protein design strategy to identify mutational hotspots and potential signatures of adaptation in these drug binding sites of Mpro. Several mutants exhibited reduced binding affinity to these drugs, out of which hotspot residues having a strong tendency to undergo positive selection were identified. The data further indicated that these anti-HCV drugs have larger footprints in the mutational landscape of Mpro and hence encompass the highest potential for positive selection and adaptation. These findings are crucial in understanding the potential structural modifications in the drug binding sites of Mpro and thus its signatures of adaptation. Furthermore, the data could provide systemic strategies for robust antiviral design and discovery against COVID-19 in the future.

Keywords: Adaptable mutations; Drug resistance; Drug tolerance; Fitness; Main protease; Protein design; SARS-CoV-2; Signatures of adaptation.

MeSH terms

  • Adaptation, Physiological / genetics*
  • Amino Acid Sequence
  • Antiviral Agents / chemistry*
  • Antiviral Agents / pharmacology
  • Binding Sites / drug effects
  • Binding Sites / genetics
  • COVID-19 Drug Treatment
  • Coronavirus 3C Proteases / antagonists & inhibitors
  • Coronavirus 3C Proteases / chemistry*
  • Coronavirus 3C Proteases / genetics
  • Coronavirus 3C Proteases / metabolism
  • Drug Design*
  • Drug Resistance, Viral / genetics*
  • Genetic Fitness / genetics
  • Hepacivirus / drug effects
  • Hepacivirus / enzymology
  • Ligands
  • Models, Molecular
  • Mutation*
  • Oligopeptides / chemistry
  • Oligopeptides / pharmacology
  • Proline / analogs & derivatives
  • Proline / chemistry
  • Proline / pharmacology
  • Reproducibility of Results
  • SARS-CoV-2 / drug effects
  • SARS-CoV-2 / enzymology*
  • SARS-CoV-2 / genetics*
  • Selection, Genetic / genetics
  • Structure-Activity Relationship


  • Antiviral Agents
  • Ligands
  • Oligopeptides
  • telaprevir
  • N-(3-amino-1-(cyclobutylmethyl)-2,3-dioxopropyl)-3-(2-((((1,1-dimethylethyl)amino)carbonyl)amino)-3,3-dimethyl-1-oxobutyl)-6,6-dimethyl-3-azabicyclo(3.1.0)hexan-2-carboxamide
  • Proline
  • 3C-like proteinase, SARS-CoV-2
  • Coronavirus 3C Proteases