Novel inhibitors of severe acute respiratory syndrome coronavirus entry that act by three distinct mechanisms

J Virol. 2013 Jul;87(14):8017-28. doi: 10.1128/JVI.00998-13. Epub 2013 May 15.

Abstract

Severe acute respiratory syndrome (SARS) is an infectious and highly contagious disease that is caused by SARS coronavirus (SARS-CoV) and for which there are currently no approved treatments. We report the discovery and characterization of small-molecule inhibitors of SARS-CoV replication that block viral entry by three different mechanisms. The compounds were discovered by screening a chemical library of compounds for blocking of entry of HIV-1 pseudotyped with SARS-CoV surface glycoprotein S (SARS-S) but not that of HIV-1 pseudotyped with vesicular stomatitis virus surface glycoprotein G (VSV-G). Studies on their mechanisms of action revealed that the compounds act by three distinct mechanisms: (i) SSAA09E2 {N-[[4-(4-methylpiperazin-1-yl)phenyl]methyl]-1,2-oxazole-5-carboxamide} acts through a novel mechanism of action, by blocking early interactions of SARS-S with the receptor for SARS-CoV, angiotensin converting enzyme 2 (ACE2); (ii) SSAA09E1 {[(Z)-1-thiophen-2-ylethylideneamino]thiourea} acts later, by blocking cathepsin L, a host protease required for processing of SARS-S during viral entry; and (iii) SSAA09E3 [N-(9,10-dioxo-9,10-dihydroanthracen-2-yl)benzamide] also acts later and does not affect interactions of SARS-S with ACE2 or the enzymatic functions of cathepsin L but prevents fusion of the viral membrane with the host cellular membrane. Our work demonstrates that there are at least three independent strategies for blocking SARS-CoV entry, validates these mechanisms of inhibition, and introduces promising leads for the development of SARS therapeutics.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Angiotensin-Converting Enzyme 2
  • Anthracenes / pharmacology*
  • Benzamides / pharmacology*
  • Blotting, Western
  • Cathepsin B / metabolism
  • Cathepsin L / antagonists & inhibitors
  • Cathepsin L / metabolism
  • DNA Primers / genetics
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • Fluorometry
  • HEK293 Cells
  • Humans
  • Immunoprecipitation
  • Luciferases
  • Oxazoles / pharmacology*
  • Peptidyl-Dipeptidase A / metabolism
  • Piperazines / pharmacology*
  • Real-Time Polymerase Chain Reaction
  • Severe acute respiratory syndrome-related coronavirus / physiology*
  • Small Molecule Libraries
  • Thiophenes / pharmacology*
  • Thiourea / pharmacology*
  • Virus Internalization / drug effects*

Substances

  • ((Z)-1-thiophen-2-ylethylideneamino)thiourea
  • Anthracenes
  • Benzamides
  • DNA Primers
  • N-((4-(4-methylpiperazin-1-yl)phenyl)methyl)-1,2-oxazole-5-carboxamide
  • N-(9,10-dioxo-9,10-dihydroanthracen-2-yl)benzamide
  • Oxazoles
  • Piperazines
  • Small Molecule Libraries
  • Thiophenes
  • Luciferases
  • Peptidyl-Dipeptidase A
  • ACE2 protein, human
  • Angiotensin-Converting Enzyme 2
  • Cathepsin B
  • Cathepsin L
  • Thiourea