title: Inhibitors of SARS-CoV entry--identification using an internally-controlled dual envelope pseudovirion assay.
abstract: Severe acute respiratory syndrome-associated coronavirus (SARS-CoV) emerged as the causal agent of an endemic atypical pneumonia, infecting thousands of people worldwide. Although a number of promising potential vaccines and therapeutic agents for SARS-CoV have been described, no effective antiviral drug against SARS-CoV is currently available. The intricate, sequential nature of the viral entry process provides multiple valid targets for drug development. Here, we describe a rapid and safe cell-based high-throughput screening system, dual envelope pseudovirion (DEP) assay, for specifically screening inhibitors of viral entry. The assay system employs a novel dual envelope strategy, using lentiviral pseudovirions as targets whose entry is driven by the SARS-CoV Spike glycoprotein. A second, unrelated viral envelope is used as an internal control to reduce the number of false positives. As an example of the power of this assay a class of inhibitors is reported with the potential to inhibit SARS-CoV at two steps of the replication cycle, viral entry and particle assembly. This assay system can be easily adapted to screen entry inhibitors against other viruses with the careful selection of matching partner virus envelopes.
authors.affiliation: Blood Systems Research Institute, University of California, San Francisco, CA 94118, USA.
authors.forename: Yanchen
authors.initials: Y
authors.lastname: Zhou
authors.forename: Juliet
authors.initials: J
authors.lastname: Agudelo
authors.forename: Kai
authors.initials: K
authors.lastname: Lu
authors.forename: David H
authors.initials: DH
authors.lastname: Goetz
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Hansell
authors.forename: Yen Ting
authors.initials: YT
authors.lastname: Chen
authors.forename: William R
authors.initials: WR
authors.lastname: Roush
authors.forename: James
authors.initials: J
authors.lastname: McKerrow
authors.forename: Charles S
authors.initials: CS
authors.lastname: Craik
authors.forename: Sean M
authors.initials: SM
authors.lastname: Amberg
authors.forename: Graham
authors.initials: G
authors.lastname: Simmons
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical / Q000379:methods* / Q000592:standards; D057166:High-Throughput Screening Assays / Q000379:methods* / Q000592:standards; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods / Q000592:standards; D045473:Severe acute respiratory syndrome-related coronavirus / Q000187:drug effects*; D014776:Virus Cultivation / Q000379:methods / Q000592:standards; D053586:Virus Internalization / Q000187:drug effects*
chemical_list: D000998:Antiviral Agents