A Four-Point Screening Method for Assessing Molecular Mechanism of Action (MMOA) Identifies Tideglusib as a Time-Dependent Inhibitor of Trypanosoma brucei GSK3β
Trypanosomes (5 x 105/mL in HMI-9 medium) were treated with pentamidine (or water solvent control) or tideglusib (5 μM, 10 μM, or DMSO solvent control) for 6 hours (37°C, 5% CO2) and then cultured in drug-free treatment as detailed in panel (A). Trypanosome densities were determined after 6 hour treatment (B), and the drugs washed from the cells by centrifugation and resuspension in fresh medium (to a density of 0.5 x 105/mL). Trypanosome proliferation in the fresh medium was monitored for 44 hours (37°C, 5% CO2) (C). Error between four separate cultures are shown. Cultures in which no cells were observed is denoted by an asterisk.