RO-275
Probe criteria
| Selectivity within target family: > 30-fold | Surpasses criterion: Automated patch clamp in HCNx overexpressing HEK cells after a single hyperpolarizing pulse of -100 mV: HCN2 (IC50 = 14.3 µM), HCN3 (IC50 = 4.6 µM, 100 fold selectivity), HCN4 (IC50 = 13.9 µM) |
| Selectivity outside target family | Surpasses criterion: Roche CEREP panel (48 targets) at 10 µM: clean |
| On target cell activity for cell-based targets: goal is < 1 µM IC50/EC50 | Surpasses criterion: Automated patch clamp in HCN1 overexpressing HEK293 cells after a single hyperpolarizing pulse of: -80 mV (IC50 = 48 nM; 97% max inhibition), -100 mV (IC50 = 46 nM; 99% max inhibition), and -120 mV (IC50 = 43 nM; 99% max inhibition) and after 50 hyperpolarizing pulses of -100 mV to simulate use dependency (UD): IC50 = 49 nM (99% max inhibition)Electrophysiology recordings in HEK293 cells (manual whole-cell patch clamp): hHCN1 (IC50 = 269 nM), rat HCN1 (IC50 = 240 nM, hHCN1 in co-expression with PEX5L (TRIP8b) (IC50 = 312 nM), hHCN1/ hHCN2 tandem (IC50 = 423 nM) |
| Control compound (100 times less potent than the probe) | Surpasses criterion: RO7239047: MC profiling results (Combination of patch clamp recordings with multi-electrode array technology) for -80, -100, -120 mV: HCN1 (IC50 = 40 µM), HCN2 and HCN4 (IC50 > 40 µM) |