| Assay Method Information | |
| | Evaluation of Virus protease (Main Protease) Inhibitory Activity |
| Description: | The Mpro/3CL protease enzyme inhibitory activity of each of the above compounds (test compounds) was evaluated using an assay kit based on FRET (product name: 3CL Protease Untagged Assay Kit, BPS Biosceience, CA, USA). Specifically, first the test compound of each concentration after the serial dilution was dispensed to a 96 well plate at 10 μl/well. Then, the 3CL protease was added at 15 ng/30 μL/well, followed by stirring at room temperature for 30 minutes to incubate. Immediately after that, 200 μM of 3CL Protease fluorogenic substrate was added at 10 μL/well. After incubation up to 4 hours under a condition of 25° C., the fluorescent emission intensity (excitation 360 nm/luminescence 460 nm) was quantified using microplate reader Cytation 5 (BioTek). From the quantified value, the IC50 value was calculated to evaluate the anti-SARS-CoV-2 enzyme inhibitory activity of each test compound. |
| Affinity data for this assay | |
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