title: Identification of a broad-spectrum antiviral small molecule against severe acute respiratory syndrome coronavirus and Ebola, Hendra, and Nipah viruses by using a novel high-throughput screening assay.
abstract: Severe acute respiratory syndrome coronavirus (SARS-CoV) and Ebola, Hendra, and Nipah viruses are members of different viral families and are known causative agents of fatal viral diseases. These viruses depend on cathepsin L for entry into their target cells. The viral glycoproteins need to be primed by protease cleavage, rendering them active for fusion with the host cell membrane. In this study, we developed a novel high-throughput screening assay based on peptides, derived from the glycoproteins of the aforementioned viruses, which contain the cathepsin L cleavage site. We screened a library of 5,000 small molecules and discovered a small molecule that can inhibit the cathepsin L cleavage of all viral peptides with minimal inhibition of cleavage of a host protein-derived peptide (pro-neuropeptide Y). The small molecule inhibited the entry of all pseudotyped viruses in vitro and the cleavage of SARS-CoV spike glycoprotein in an in vitro cleavage assay. In addition, the Hendra and Nipah virus fusion glycoproteins were not cleaved in the presence of the small molecule in a cell-based cleavage assay. Furthermore, we demonstrate that the small molecule is a mixed inhibitor of cathepsin L. Our broad-spectrum antiviral small molecule appears to be an ideal candidate for future optimization and development into a potent antiviral against SARS-CoV and Ebola, Hendra, and Nipah viruses.
  
  
  OBJECTIVE
  We developed a novel high-throughput screening assay to identify small molecules that can prevent cathepsin L cleavage of viral glycoproteins derived from SARS-CoV and Ebola, Hendra, and Nipah viruses that are required for their entry into the host cell. We identified a novel broad-spectrum small molecule that could block cathepsin L-mediated cleavage and thus inhibit the entry of pseudotypes bearing the glycoprotein derived from SARS-CoV or Ebola, Hendra, or Nipah virus. The small molecule can be further optimized and developed into a potent broad-spectrum antiviral drug.
authors: Department of Microbiology and Immunology, College of Medicine, University of Illinois at Chicago, Chicago, Illinois, USA.
authors: Hatem A
authors: HA
authors: Elshabrawy
authors: Jilao
authors: J
authors: Fan
authors: Christine S
authors: CS
authors: Haddad
authors: Kiira
authors: K
authors: Ratia
authors: Christopher C
authors: CC
authors: Broder
authors: Michael
authors: M
authors: Caffrey
authors: Bellur S
authors: BS
authors: Prabhakar
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D056668:Cathepsin L / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D029043:Ebolavirus / Q000187:drug effects* / Q000378:metabolism; D045404:Hendra Virus / Q000187:drug effects* / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D045405:Nipah Virus / Q000187:drug effects* / Q000378:metabolism; D045473:Severe acute respiratory syndrome-related coronavirus / Q000187:drug effects* / Q000378:metabolism; D054852:Small Molecule Libraries / Q000494:pharmacology*; D014759:Viral Envelope Proteins / Q000378:metabolism; D014777:Virus Diseases / Q000201:enzymology / Q000821:virology
chemical_list: D000998:Antiviral Agents; D054852:Small Molecule Libraries; D014759:Viral Envelope Proteins; D056668:Cathepsin L