Development and validation of a high-throughput screen for inhibitors of SARS CoV and its application in screening of a 100,000-compound library.
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-microM concentration, 3 compounds with selective indexes (SI50) of > 53 were discovered.
Severson
William E
WE
Department of Biochemistry and Molecular Biology, Southern Research Institute, Birmingham, AL 35205, USA.
Shindo
Nice
N
Sosa
Mindy
M
Fletcher
Thomas
T
White
E Lucile
EL
Ananthan
Subramaniam
S
Jonsson
Colleen B
CB
D000818:Animals; D000998:Antiviral Agents / Q000032:analysis* / Q000737:chemistry / Q000494:pharmacology*; D002452:Cell Count; D002522:Chlorocebus aethiops; D020650:Combinatorial Chemistry Techniques / Q000379:methods*; D004121:Dimethyl Sulfoxide; D004353:Drug Evaluation, Preclinical; D023381:Endpoint Determination; D020128:Inhibitory Concentration 50; D049449:Luminescence; D015203:Reproducibility of Results; D045473:Severe acute respiratory syndrome-related coronavirus / Q000187:drug effects*; D014709:Vero Cells
D000998:Antiviral Agents; D004121:Dimethyl Sulfoxide