CHEMBL5738502

Kinase Inhibition: Kinase assay: After the buffer was prepared, the enzyme was mixed with different concentrations of the compound prepared by pre-diluting, and allowed to stand at room temperature for 30 minutes, with each concentration in duplicate. The corresponding substrate and ATP were added and reacted at room temperature for 60 minutes (wherein negative and positive controls were set). After the reaction was completed, the antibody was added for detection. After incubation at room temperature for 60 minutes, Evnvision detection was performed, and data were collected. The enzyme activities in the presence of varying concentration of the compounds of the present invention were determined by Evnvision microplate reader, and the inhibitory activities of the compounds at different concentrations on the enzyme activity were calculated. Then, the inhibitory activities of the compounds at different concentrations on the enzyme activity were fitted to the four-parameter equation according to the Graphpad 5.0 software, and the IC50 values were calculated.