ACBI3
Probe criteria
| Inhibitor/agonist potency: goal is < 100 nM (IC50, KD) | Surface plasmon resonance (SPR): KRASG12D GDP (KD = 5 ± 1 nM; n = 3), KRASG12V GDP (KD = 4 ± 1 nM; n = 3) Fluorescence polarization (FP): VCB+KRASG12D (KD = 4 ± 1 nM; n = 3) |
| Selectivity within target family: > 30-fold | Whole cell proteomics MS analysis of GP2d cells: selective degradation of KRAS. HRAS (log2 fold change -0.0006, -logP 0.001) and NRAS (log2 fold change -0.12, -logP 0.52) levels are not significantly affected. |
| Selectivity outside target family | Whole cell proteomics MS analysis of GP2d cells: clean |
| On target cell activity for cell-based targets: goal is < 1 µM IC50/EC50 | Cellular KRASG12D degradation (24 h, GP5d cells): DC50 = 2 nM; Cellular KRASG12V degradation (24 h, SW620 cells): DC50 = 7 nM (Assay: capillary electrophoresis using the following antibodies: KRASG12D, KRASG12V, normalized by GAPDH) Cellular proliferation (5 days): GP5d cells (IC50 = 5 nM), SW620 cells (IC50 = 15 nM) (CellTiterGlo assay) |
| Control compound (100 times less potent than the probe) | cis-ACBI3: Cellular KRASG12D degradation (24 h, GP5d cells): DC50 > 1 µM; Cellular proliferation (5 days): GP5d cells (IC50 > 1 µM) |