Figure 3.

In vitro assay. (a) Selected structure of active compound, N-(3-(2-ethyl-1H-imidazol-1-yl)propyl)-5-(4-fluorophenyl)pyrazolo[1,5-a]pyrimidin-7-amine (cpd1) and dose–response curve fit of cpd1 and reference SB216763. (b) Competitive inhibition by cpd1 and SB216873. Compounds were tested using 4-point 2-fold serial dilution, starting from 1 µM or 0.25 µM at 5 different concentration of ATP (200, 100, 50, 33.3, and 25 µM). (c) Reversibility test. GSK3β enzyme was pre-incubated with 1 µM (>20-fold of IC₅₀) cpd1 for 30 min and diluted with buffer into indicated concentrations.