Hit Confirmation

Based on primery screening results the most perspective molecules could be tested on CPE assay to measuse general antiviral activity. Also this molecules could be screened on enzymatic assays for IC50 measure.

CPE assay

Cell line: Vero E6

Virus (origin/strain): SARS-Cov-2

Treatment: 4 days

Infection: 100 and/or 1000 TCID50

Detection: MTT and/or microscope

Real-time PCR for SARS-Cov-2 RNA: yes, Ct

Cytotoxicity: yes

Format: 96-well, single compound test

Reference: hydroxychloroquine

IC50

Screening the most perspectives compounds for measuring its IC50 on one of enzymatic assays: 3CL, PLpro, RdRp, ACE2-S1.