Structural Basis for Inhibiting Porcine Epidemic Diarrhea Virus Replication with the 3C-Like Protease Inhibitor GC376

Viruses. 2020 Feb 21;12(2):240. doi: 10.3390/v12020240.

Abstract

Porcine epidemic diarrhea virus (PEDV), being highly virulent and contagious in piglets, has caused significant damage to the pork industries of many countries worldwide. There are no commercial drugs targeting coronaviruses (CoVs), and few studies on anti-PEDV inhibitors. The coronavirus 3C-like protease (3CLpro) has a conserved structure and catalytic mechanism and plays a key role during viral polyprotein processing, thus serving as an appealing antiviral drug target. Here, we report the anti-PEDV effect of the broad-spectrum inhibitor GC376 (targeting 3Cpro or 3CLpro of viruses in the picornavirus-like supercluster). GC376 was highly effective against the PEDV 3CLpro and exerted similar inhibitory effects on two PEDV strains. Furthermore, the structure of the PEDV 3CLpro in complex with GC376 was determined at 1.65 Å. We elucidated structural details and analyzed the differences between GC376 binding with the PEDV 3CLpro and GC376 binding with the transmissible gastroenteritis virus (TGEV) 3CLpro. Finally, we explored the substrate specificity of PEDV 3CLpro at the P2 site and analyzed the effects of Leu group modification in GC376 on inhibiting PEDV infection. This study helps us to understand better the PEDV 3CLpro substrate specificity, providing information on the optimization of GC376 for development as an antiviral therapeutic against coronaviruses.

Keywords: PEDV 3CLpro; complex; coronavirus; crystal structure; inhibitor.

MeSH terms

  • Animals
  • Antiviral Agents / chemistry
  • Antiviral Agents / metabolism
  • Antiviral Agents / pharmacology*
  • Catalytic Domain
  • Chlorocebus aethiops
  • Crystallography, X-Ray
  • Models, Molecular
  • Peptide Hydrolases / chemistry*
  • Peptide Hydrolases / metabolism
  • Porcine epidemic diarrhea virus / drug effects*
  • Porcine epidemic diarrhea virus / enzymology
  • Porcine epidemic diarrhea virus / physiology
  • Protease Inhibitors / chemistry
  • Protease Inhibitors / metabolism
  • Protease Inhibitors / pharmacology*
  • Pyrrolidines / chemistry
  • Pyrrolidines / metabolism
  • Pyrrolidines / pharmacology*
  • Substrate Specificity
  • Transmissible gastroenteritis virus / enzymology
  • Vero Cells
  • Virus Replication / drug effects

Substances

  • Antiviral Agents
  • GC376
  • Protease Inhibitors
  • Pyrrolidines
  • Peptide Hydrolases