Donated Chemical Probes

Target	BRD2 BRD3 BRD4 BRDT
Targeted domain	BD2
Mode of action	Inhibitor
Control
Orthogonal probe	GSK620
Recommended cellular usage concentration	≤ 10 µM
In vivo use	Yes
Synonyms	iBET-BD2, Compound 59
Donated by	GlaxoSmithKline

Inhibitor/agonist potency: goal is < 50 nM (IC₅₀, K_D)	Surpasses criterion: :BET mutant TR-FRET assay: BRD2 (BD1) pIC₅₀ = 5.0; (BD2) 6.6 ± 0.2 (n= 9) BRD3 (BD1) pIC₅₀ = 4.4 ± 0.01 (n = 2); (BD2) 7.0 ± 0.08 (n = 8) BRD4 (BD1) pIC₅₀ = 4.2 ± 0.44 (n = 3); (BD2) 7.3 ± 0.13 (n = 12); 1260-fold BRDT (BD1) pIC₅₀ < 4.3 (n = 8); (BD2) 6.7 ± 0.25 (n = 8) BROMOScan (DiscoverX): BRD2 (BD1): K_d = 1621 nM; (BD2) 35 nM; 64-fold BRD3 (BD1): K_d= 2082 nM; (BD2) 32 nM ; 63-fold BRD4 (BD1): K_d =769 nM; (BD2) 9 nM; 85-fold BRDT (BD1): K_d = 2454 nM; (BD2) 15 nM; 164-fold SPR BRD4 (BD1): K_d > 10 µM; BRD4 (BD2) K_d = 30 nM
Selectivity within target family: > 30-fold	Surpasses criterion:: BROMOScan (DiscoverX) (40 tested): clean
Selectivity outside target family	Selectivity screen (48 targets tested): clean Clean PDSP scan exept for one off-target: HTR1D Ki = 3438.75 nM
On target cell activity for cell-based targets: goal is < 1 µM IC₅₀/EC₅₀	Surpasses criterion: Lipopolysaccharide (LPS)-stimulated human whole blood assay: MCP-1 (pIC₅₀ = 6.5 ± 0.1 (n=6)) LPS-PBMC assay (MCP-1) pIC₅₀ = 7.5 ± 0.3 (n = 4)