Development and Validation of a High-Throughput Screen for Inhibitors of SARS CoV and Its Application in Screening of a 100,000-Compound Library

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 (SI 50 ) of > 53 were discovered. (Journal of Biomolecular Screening 2007:33-40)

New Substructure Filters for Removal of Pan Assay Interference Compounds (PAINS) from Screening Libraries and for Their Exclusion in Bioassays

“…These include certain guanidines, 74 and certain arysulfonamides (it seems unlikely that the arylsulfonamides involved would be reactive enough to form a Meisenheimer complex with protein thiols and the reasons for their appearance in these screening hit sets remains unknown). [21][22][23][24]28,130 Some dye-like compounds with conjugated and often charged nitrogen atoms also appear in some of the screening literature already cited. 74, 168 We find that several of these also interfere in our assays, which is unsurprising considering the results shown in Table 9.…”

Acoustic Droplet Ejection Applications for High-Throughput Screening of Infectious Agents

“…In that time, more than 9 million compounds have been screened against 36 infectious agents, at both the BSL-2 and the BSL-3 level. [9][10][11][12][13][14][15][16][17] Many assays were adapted to the automation platform and screened successfully. But there were assays that could only achieve semi-high throughput due to the technical limitations of conventional liquid handling.…”

Screening of the antiviral activity in the range of C5 and N3 substituted 4-thiazolidinone derivatives

“…The study involved the standard assay protocols regarding coronavirus SARS [24], herpes viruses (Herpes simplex virus 1 and 2 (HSV-1, HSV-2)), varicella zoster virus (VZV), Epstein-Barr virus (EBV), cytomegalovirus (HCMV) [25], hepatitis C (HCV) [26], and B (HBV) viruses [27], as well as vaccinia virus (Vaccinia, Cowpox) and influenza viruses of type A and type B, adeno-and rhinovirus; viruses of biological weapons [28] -Dengue fever virus, yellow fever virus, Takaribe virus, Rift Valley Fever vitus, Venezuelan equine encephalitis virus (VEE), measles virus, pig influenza virus (PIV), respiratory syncytial virus (RSV). The results obtained (Tab.…”