A-1211212
Probe criteria
| Inhibitor/agonist potency: goal is < 100 nM (IC50, KD) | Surpasses criterion: BCL2: IC50 = 1.4 nM (Time-resolved fluorescence energy transfer (TR-FRET)) |
| Selectivity within target family: goal is > 30-fold | Surpasses criterion: BCL2L1: IC50 = 753 nM (TR-FRET); MCL1: IC50 = 2050 nM (TR-FRET) |
| Selectivity outside target family | Closest off target for the probe at 10 µM is the CACNA1S (Ca2+ channel (L, dihydropyridine site)) with 63.2 % inhibition (Eurofins-Cerep screen). One off-target in PDSP scan: TMEM97 (Ki = 1491.08 nM)KINOMEScan with 468 targets at 1 µM: Closest off-targets of primary data (% Ctrl) are HIPK4 (4.4), MAPK12 (47) |
| On target cell activity for cell-based targets: goal is < 1 µM IC50/EC50 | Surpasses criterion: Cellular selectivity demonstrated – induces apoptosis in BCL2 dependent cell lines but not MCL1 or BCL2L1 dependent cell lines. EC50 = 6 nM in CellTiter-Glo assays (10 % HS, 48 hr) in BCL2 dependent RS4;11 cells (EC50 > 5000 nM in BCL2L1 dependent H146 cells (10 % HS, 48 hr) and EC50 > 1000 nM in MCL1 dependent AMO-1 cells (10 % HS, 24 hr)) |
| Control compound (100 times less potent than the probe) | Surpasses criterion: A-1210227 >100-fold less potent in binding and cellular assays (BCL2: IC50 = 296 nM (TR-FRET)) Closest off-target in the PDSP (43) scan is TMEM97 (Ki = 8371.44 nM).KINOMEScan with 468 targets at 1 µM: clean |