Article
High throughput screening identifies inhibitors of the SARS coronavirus main proteinase
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Authors
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Jan E. Blanchard
- McMaster High Throughput Screening Laboratory, Department of Biochemistry, McMaster University, Hamilton, ON, Canada
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Nadine H. Elowe
- McMaster High Throughput Screening Laboratory, Department of Biochemistry, McMaster University, Hamilton, ON, Canada
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Pascal D. Fortin
- Departments of Microbiology and Biochemistry, University of British Columbia, Vancouver, BC, Canada
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Carly Huitema
- Departments of Microbiology and Biochemistry, University of British Columbia, Vancouver, BC, Canada
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Jonathan D. Cechetto
- McMaster High Throughput Screening Laboratory, Department of Biochemistry, McMaster University, Hamilton, ON, Canada
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Lindsay D. Eltis
- Departments of Microbiology and Biochemistry, University of British Columbia, Vancouver, BC, Canada
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Eric D. Brown
- McMaster High Throughput Screening Laboratory, Department of Biochemistry, McMaster University, Hamilton, ON, Canada
| Published: | May 26, 2004 |
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Outline
Text
The main proteinase of the SARS coronavirus, 3CLpro, is an attractive target for therapeutics against SARS owing to its fundamental role in viral replication. We sought to identify novel inhibitors of 3CLpro through a high throughput screening campaign to advance the development of appropriate therapies in the treatment of SARS. 3CLpro was cloned, expressed and purified from the Tor2 isolate. A quenched fluorescence resonance energy transfer assay was developed with a nine amino acid peptide labeled with 2-aminobenzoyl and nitrotyrosine on opposing sides of the proteinase cleavage site. This assay was used to screen 3CLpro against 50,000 drug-like small molecules on a fully automated system. The screening campaign yielded 572 primary hits; through a series of virtual and experimental filters, this number was reduced to 5 novel small molecules that show potent and selective inhibitory activity towards SARS-CoV 3CLpro.
