Development and Validation of a High-Throughput Screen for Inhibitors of SARS CoV and Its Application in Screening of a 100,000-Compound Library
Identifieur interne : 000F16 ( Main/Exploration ); précédent : 000F15; suivant : 000F17Development and Validation of a High-Throughput Screen for Inhibitors of SARS CoV and Its Application in Screening of a 100,000-Compound Library
Auteurs : William E. Severson [États-Unis] ; Nice Shindo ; Mindy Sosa [États-Unis] ; Thomas Fletcher [États-Unis] ; E. Lucile White [États-Unis] ; Subramaniam Ananthan [États-Unis] ; Colleen B. Jonsson [États-Unis]Source :
- Journal of Biomolecular Screening [ 1087-0571 ] ; 2007-02.
English descriptors
- Teeft :
- Accurate indication, Active compound, Assay, Assay media, Assay performance, Average value, Beckman coulter biomek, Biomolecular, Biomolecular screening, Calpain, Calpain inhibitor, Cell control, Cell control signal, Cell controls, Cell number, Cell viability, Compound, Coronavirus, Cytopathic, Cytopathic effect, Dehydrogenase inhibitors, Different days, Dmso, Endpoint, Endpoint reagent, Excellent candidates, Further evaluation, Highest concentration, Hong kong, Incubation time, Inhibitor, Inhibitory effects, Luminescence, Luminescent signal, Major outbreak, Microsource, Microsource library, Microsource spectrum library, Molecular biology, Ninds, Ninds library, Plate reader, Positive control, Positive control drug, Potential inhibitors, Prestwick, Prestwick library, Reagent, Relative luminescence, Respiratory syndrome, Respiratory syndrome coronavirus, Sars, Sars coronavirus, Screening, Selective index, Serial dilutions, Several compounds, Several drawbacks, Small compound libraries, Standard deviation, Syndrome, Tetrazolium compound, Validation, Vero, Viable cells, Viral stock.
Abstract
The authors have developed a high-throughput screen (HTS) that allows for the identification of potential inhibitors of the severe acute respiratory syndrome coronavirus (SARS CoV) from large compound libraries. The luminescent-based assay measures the inhibition of SARS CoV–induced cytopathic effect (CPE) in Vero E6 cells. The assay was validated in 96-well plates in a BSL3 containment facility. The assay is sensitive and robust, with Z values > 0.6, signal to background (S/B) > 16, and signal to noise (S/N) > 3. The assay was further validated with 2 different diversity sets of compounds against the SARS CoV. The “hit” rate for both libraries was approximately 0.01%. The validated HTS assay was then employed to screen a 100,000-compound library against SARS CoV. The hit rate for the library in a single-dose format was determined to be approximately 0.8%. Screening of the 3 libraries resulted in the identification of several novel compounds that effectively inhibited the CPE of SARS CoV in vitro—compounds which will serve as excellent lead candidates for further evaluation. At a 10-μM concentration, 3 compounds with selective indexes (SI50) of > 53 were discovered.
Url:
DOI: 10.1177/1087057106296688
Affiliations:
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The luminescent-based assay measures the inhibition of SARS CoV–induced cytopathic effect (CPE) in Vero E6 cells. The assay was validated in 96-well plates in a BSL3 containment facility. The assay is sensitive and robust, with Z values > 0.6, signal to background (S/B) > 16, and signal to noise (S/N) > 3. The assay was further validated with 2 different diversity sets of compounds against the SARS CoV. The “hit” rate for both libraries was approximately 0.01%. The validated HTS assay was then employed to screen a 100,000-compound library against SARS CoV. The hit rate for the library in a single-dose format was determined to be approximately 0.8%. Screening of the 3 libraries resulted in the identification of several novel compounds that effectively inhibited the CPE of SARS CoV in vitro—compounds which will serve as excellent lead candidates for further evaluation. At a 10-μM concentration, 3 compounds with selective indexes (SI50) of > 53 were discovered.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Alabama</li></region></list><tree><noCountry><name sortKey="Shindo, Nice" sort="Shindo, Nice" uniqKey="Shindo N" first="Nice" last="Shindo">Nice Shindo</name></noCountry><country name="États-Unis"><region name="Alabama"><name sortKey="Severson, William E" sort="Severson, William E" uniqKey="Severson W" first="William E." last="Severson">William E. Severson</name></region><name sortKey="Ananthan, Subramaniam" sort="Ananthan, Subramaniam" uniqKey="Ananthan S" first="Subramaniam" last="Ananthan">Subramaniam Ananthan</name><name sortKey="Fletcher, Thomas" sort="Fletcher, Thomas" uniqKey="Fletcher T" first="Thomas" last="Fletcher">Thomas Fletcher</name><name sortKey="Jonsson, Colleen B" sort="Jonsson, Colleen B" uniqKey="Jonsson C" first="Colleen B." last="Jonsson">Colleen B. Jonsson</name><name sortKey="Sosa, Mindy" sort="Sosa, Mindy" uniqKey="Sosa M" first="Mindy" last="Sosa">Mindy Sosa</name><name sortKey="White, E Lucile" sort="White, E Lucile" uniqKey="White E" first="E. Lucile" last="White">E. Lucile White</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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