ERKi
Probe criteria
| Inhibitor/agonist potency: goal is < 100 nM (IC50, Kd) | Surpasses criteria: MAPK1/3 potency is < 10 nM in MSD and Invitrogen assays |
| Selectivity within target family: > 30-fold | Surpasses criteria: > 30-fold against Invitrogen kinase panel. Closest off targets: GSK3A = 0.21 µM, SGK1 = 0.27 µM, GSK3B = 0.54 µM, GRK2 (ADRBK1) = 0.59 µM, TBK1 = 0.77 µM, CDK2 = 0.79 µM, STK26 (MST4) = 0.88 µM, CLK2 = 0.94 µM, PRKG2 = 0.95 µM, CDK5 = 1.2 µM, ROCK1 = 1.2 µM, ROCK2 = 1.4 µM, MARK2 = 1.4 µMNanoBRET results: GRK2 (IC50 > 20µM), from SGC Frankfurt: GSK3A (EC50 = 13 ± 1 µM), SGK1 (EC50 > 50 µM) , GSK3B (EC50 = 5.2 ± 1.9 µM) TBK1 (EC50 = 22 ± 4.8 µM) |
| Selectivity outside target family | Profiled in a panel of 116 enzymes and receptors at MDS Pharma/Taiwan: only one unconfirmed hit, PDE3A (61% @ 10 µM) is > 1000-fold MAPK1/3 potency; Clean PDSP scan |
| On target cell activity for cell-based targets: goal is < 1 µM IC50/EC50 | Surpasses criteria: 280 nM pRSK in A375SM cellsNanoBRET assay (SGC Frankfurt, n= 3): MAPK1 (EC50 = 95 ± 19 nM), MAPK3 (EC50 = 32 ± 11 nM) |
| Control compound (100 times less potent than the probe) | Surpasses criteria: The negative control is 1000-fold less potent biochemically and shows no activity in cells @ 10µM; PDSP scan shows weak off-target activity for HTR2B (pKi = 6.27).NanoBRET results (SGC Frankfurt): GSK3A, SGK1, GSK3B, GRK2 and TBK1 (all EC50 > 50µM), MAPK3 (EC50 = 6.7 ± 1 µM), MAPK1 (EC50 =18 ± 2.7 µM) |