GSK789
Probe criteria
| Inhibitor/agonist potency: goal is < 50 nM (IC50, KD) | Surpasses criterion: :BET mutant TR-FRET assay:
BRD2 (BD1) pIC50 = 7.0; (BD2) 4.7; 200-fold BRD3 (BD1) pIC50 =6.8; (BD2) 4.7; 125-fold BRD4 (BD1) pIC50 = 7.5 ± 0.26 (n = 4); (BD2) 4.5 ± 0.09 (n = 2); 1000-fold BRDT (BD1) pIC50 = 6.2; (BD2) 4.9; 20-fold BROMOScan (DiscoverX): BRD2(1): pKd = 7.7; (BD2) 4.6; 1200-fold; BRD3(1): pKd= 7.8; (BD2) 4.7; 1200-fold; BRD4(1): pKd =7.7; (BD2) 4.5; 1300-fold; BRDT(1): pKd = 7.7; (BD2) 4.7; 1000-fold Chemoproteomics (full-length protein): BRD2(1): pKd = 8.4; (BD2) < 4.6; < 6000-fold; BRD3(1): pKd=8.4; (BD2) 5.2; 1600-fold; BRD4(1): pKd =8.2; (BD2) <4.7; > 3000-fold; BRDT(1): pKd =8.0; (BD2) < 4.5; > 3000-fold |
| Selectivity within target family: > 30-fold | Surpasses criterion:BROMOScan (DiscoverX) (34 tested): selective for the BD1 domain of BET proteins with the exception of the following two hits: TAF1 BD2 (pKd = 7.3) and TAF1L BD2 (pKd = 6.4). But In house TAF(1) FRET pIC50 = 5.0 ± 0.1, n = 3 (BRD4 BD1 selectivity = 500-fold). |
| Selectivity outside target family | Clean in-house panel (40 targets, data not shown)Clean PDSP scan except for one off-target: DRD4 Ki = 1013.12 nM |
| On target cell activity for cell-based targets: goal is < 1 µM IC50/EC50 | Surpasses criterion: Potent antiproliferative activity in various cancer cell lines: MV-4−11 (IC50 = 125 nM); HL60 (IC50 = 390 nM); THP-1 (IC50 = 258 nM) Lipopolysaccharide (LPS)-stimulated human whole blood assay: MCP-1 (IC50 = 0.67 µM ± 0.03 (n=6)); TNFα (IC50 = 0.87 µM ± 0.19 (n=5)); IL-6 (IC50 = 3.55 µM ±1.74 (n=5)) LPS-PBMC assay (MCP-1) pIC50 = 7.7 |
| Control compound (100 times less potent than the probe) | Surpasses criterion: GSK791 BET mutant TR-FRET assay:
BRD2 (BD1) pIC50 = 5.6; (BD2) 4.8 BRD3 (BD1) pIC50 = 5.4; (BD2) 4.7 BRD4 (BD1) pIC50 = 5.8; (BD2) 4.5 BRDT (BD1) pIC50 = 5.2; (BD2) 5.1 Clean PDSP scan except for one off-target: GABAA/BZP (Ki = 2539.22 nM) |