Potency comparison of FLT3 inhibitors in biochemical and cellular assays
| Compound . | FLT3 activity (IC50, nM) . | Human leukemia cell line (IC50, nM) . | BaF3 cells with FLT3 mutants (IC50, nM) . | |||||||
|---|---|---|---|---|---|---|---|---|---|---|
| WT . | D835Y . | MV4-11 . | Molm-14 . | WT . | ITD . | ITD/N676D . | ITD/F691L . | D835Y . | D835Y/N676D . | |
| G-749 | 0.4 ± 0.2 | 0.6 ± 0.2 | 3.5 ± 0.9 | 7.5 ± 0.3 | 6.1 ± 0.4 | 11.8 ± 3.7 | 21.4 ± 1.1 | 38.1 ± 6.6 | 3.1 ± 0.5 | 2.1 ± 0.7 |
| AC220 | 8.8 ± 1.4 | 28.2 ± 2.1 | 1.1 ± 0.4 | 1.3 ± 0.5 | 9.0 ± 1.8 | 3.7 ± 0.8 | 36.2 ± 4.0 | 194.2 ± 28 | 52.4 ± 4.7 | 149.7 ± 27.0 |
| PKC412 | 15.4 ± 3.5 | 24.2 ± 3.1 | 18.5 ± 2.8 | 21.5 ± 1.5 | 28.9 ± 3.0 | 21.6 ± 1.1 | 128.7 ± 29 | 16.1 ± 4.7 | 11.4 ± 1 | 17.3 ± 3.7 |
| Compound . | FLT3 activity (IC50, nM) . | Human leukemia cell line (IC50, nM) . | BaF3 cells with FLT3 mutants (IC50, nM) . | |||||||
|---|---|---|---|---|---|---|---|---|---|---|
| WT . | D835Y . | MV4-11 . | Molm-14 . | WT . | ITD . | ITD/N676D . | ITD/F691L . | D835Y . | D835Y/N676D . | |
| G-749 | 0.4 ± 0.2 | 0.6 ± 0.2 | 3.5 ± 0.9 | 7.5 ± 0.3 | 6.1 ± 0.4 | 11.8 ± 3.7 | 21.4 ± 1.1 | 38.1 ± 6.6 | 3.1 ± 0.5 | 2.1 ± 0.7 |
| AC220 | 8.8 ± 1.4 | 28.2 ± 2.1 | 1.1 ± 0.4 | 1.3 ± 0.5 | 9.0 ± 1.8 | 3.7 ± 0.8 | 36.2 ± 4.0 | 194.2 ± 28 | 52.4 ± 4.7 | 149.7 ± 27.0 |
| PKC412 | 15.4 ± 3.5 | 24.2 ± 3.1 | 18.5 ± 2.8 | 21.5 ± 1.5 | 28.9 ± 3.0 | 21.6 ± 1.1 | 128.7 ± 29 | 16.1 ± 4.7 | 11.4 ± 1 | 17.3 ± 3.7 |
Biochemical kinase assay for FLT3-WT and FLT3-D835Y obtained from Carna Bioscience Inc. was performed in time-resolved fluorescence resonance energy transfer in series concentration of G-749, AC220, and PKC412, and their IC50 values were calculated. For the determination of IC50 values in human leukemia cell lines, the MV4-11 and Molm-14 cells were treated with test inhibitors, and cell viability was measured. The BaF3 model cell expressing double FLT3 mutants, FLT3-ITD/N676D, FLT3-ITD/D835Y, and FLT3-D835Y/N676D, and single mutants, FLT3-ITD and FLT3-D835Y, were treated with test inhibitors, and cell viability was measured.