BAY-277
Probe criteria
| Inhibitor/agonist potency: goal is < 100 nM (IC50, Kd) | Surpasses criterion: Biochemical assay: hMETAP2 (IC50 = 5.8 nM), mouse METAP2 (IC50 = 5.9 nM) |
| Selectivity within target family: > 30-fold | Proteomics: MetAP2 is the only protein among ~5000 detected proteins in HUVEC cells which is highly and significantly reduced |
| Selectivity outside target family | Eurofins Panlabs panel (76 targets) at 10 µM: Closest off-targets (% inh.): DRD3 (90), HRH3 (81), ADRA2C (79)Eurofins kinase panel at 1 µM: clean (25 kinases > 10 % inh., max 29% inh.)Closest off-targets in the PDSP panel (43 targets, at 10 µM) are HRH3 (Ki = 72.84 nM), CHRM5 (Ki = 567.76 nM), CHRM4 (Ki = 631.53 nM), CHRM2 (Ki = 676.76 nM), SIGMAR1 (Ki =866.55 nM). |
| On target cell activity for cell-based targets: goal is < 1 µM IC50/EC50 | Surpasses criterion: Degradation in HT1080 cells (Capillary electrophoresis) DC50= 8.93 nM, Dmax = 88.5 %; Degradation in HUVEC cells (Western Blot): DC50= 0.2 nM, Dmax = 94 %;2D HUVEC cells proliferation assay: IC50 = 12 nM Cellular efficacy shown in 14-3-3 methionine peptidase assay: MetAP2 degradation results in strong inhibition of peptidase activity; The mode of action was shown by Proteasome-dependent degradation, rescue experiments with CRBN binder, target binder and proteasome inhibiton. |
| Control compound (100 times less potent than the probe) | BAY-8805 (E3-blocked MetAP2 bifunction): Biochemical assay: hMETAP2 (IC50 = 5.7 nM), Degradation in HT1080 cells (capillary electrophoresis): DC50 > 1 µM; Degradation in HUVEC cells (Western Blot): DC50 > 100 nM; 2D HUVEC cells proliferation assay: IC50 = 450 nMEurofins Panlabs panel (76 targets) at 10 µM: Closest off-targets (% inh.): HRH3 (85), DRD3(76), ADRA2C (62); Eurofins kinase panel at 1 µM: clean (44 kinases > 10 % inh., max 26% inh.)Closest off-targets in the PDSP panel (43 targets, at 10 µM) are HRH3 (Ki = 73.22 nM), DRD3 (Ki = 525.3 nM), CHRM4 (Ki = 893.72 nM), SIGMAR1 (Ki =1142.33 nM). |