BAY 1816032
Probe criteria
| Inhibitor/agonist potency: goal is < 100 nM (IC50, Kd) | Biochemical assay: IC50 = 6.5 nM (low ATP), 1.9 μM (high ATP (2mM)) (w/o preinc.); TR-FRET: EC50 = 6.1 ± 2.5 nM at 10 μM ATP; SPR: Kd = 2.1 nM; ePCA binding competition (equilibrium binding assay): IC50 = 1 ± 0.5 nM; Kinome scan: Kd = 3.3 nM |
| Selectivity within target family: > 30-fold | DiscoverX (403 kinases at 100 nM and 1 μM): Closest off-targets (Kd [nM]): STK10 (57) (17-fold), CDC42BPG (850), DDR1 (2300); all > 250-fold selective |
| Selectivity outside target family | Eurofins-Panlabs Screen (89 targets at 10 μM): Closest off-targets [% inh.] are human Adenosine transporter (71, IC50 = 370 nM; 51-fold), ADORA2A (67), GABAA Chloride Channel TBOB receptor (60), PTGS1 (54), sodium channel site 2 (53) and Phosphodiesterase PDE4 (52) Clean PDSP scan (44 targets) at 10 µM |
| On target cell activity for cell-based targets: goal is < 1 µM IC50/EC50 | Surpasses criterion:Inhibits tumor cell proliferation with a median IC50 = 1.4 μM in a very uniform manner (range of measured IC50's for different tumor cell lines: between 100 nM and 58 μM).Abrogation of histone H2A-Thr120 phosphorylation, a substrate of BUB1 kinase, in nocodazole arrested HeLa cells after 1 hour compound incubation: IC50 = 29 ± 23 nM; Formation of H2A-Thr120: IC50 = 43 nM |
| Control compound (100 times less potent than the probe) | BAY-283: IC50 = 1.71 µM (low ATP, 260-fold less potent than probe), IC50 > 20 μM (2mM ATP); In-house kinase panel (25 targets: all IC50 > 20 μM) Closest off-targets in the PDSP scan (44 targets) at 10 µM are TMEM97 (Ki = 655.24 nM), SIGMAR1 (Ki = 1340.29 nM), SLC6A3 (Ki = 1626.67 nM) and SLC6A2 (Ki = 2744.73 nM). |