Donated Chemical Probes

Target	IRAK4
Targeted domain	Kinase domain (active site)
Mode of action	Inhibitor (type 1, ATP competitive)
Control	GNE-6689
Recommended cellular usage concentration	1 µM
In vivo use	Yes
Synonyms	Compound 19, IRAK4i-2
Donated by	Genentech

Inhibitor/agonist potency: goal is < 100 nM (IC₅₀, K_d)	Surpasses criterion: Biochemical assay (FRET): Ki = 1.4 ± 0.32 nM (n = 30); Maximal inhibition: 82 ± 9.2 %
Selectivity within target family: > 30-fold	Kinase panel (221) at 1 µM: Closest off-targets (IC50 [nM]) are FLT3 (177), JAK1 (282), JAK2 (486), LRRK2 (198), MAP4K4 (680), MINK1 (879), MAP3K7- TAB1 (TAK1-TAB1) (966), NTRK1 (313), NTRK2 (259) NanoBRET assay (SGC Frankfurt) (EC₅₀ [µM]; n = 3): FLT3 (> 50), JAK1 (> 50), JAK2 (> 50), LRRK2 (5.2 ± 0.74), NTRK1 (11 ± 0.99), NTRK2 (5.6 ± 1.2)
Selectivity outside target family	CEREP panel (36) at 1 µM: Off-targets with > 50% inhibition are TACR1 (NK1), HTR2B, ACHE PDSP scan (44 targets) at 10 µM clean except for HTR2B (Ki = 509.45 nM)
On target cell activity for cell-based targets: goal is < 1 µM IC₅₀/EC₅₀	Surpasses criterion: NanoBRET: EC₅₀ = 3 ± 0.3 nM (SGC Frankfurt) IL-6 human whole blood assay: IC₅₀ = 190 ± 79 nM (n=10) IFNα human whole blood assay: IC₅₀ = 290 nM
Control compound (100 times less potent than the probe)	GNE-6689: NanoBRET assay (SGC Frankfurt) (EC₅₀ [µM]; n =3): IRAK4 (9.5 ± 0.54), FLT3 (3.6 ± 0.74), JAK1 (> 50), JAK2 (> 50), LRRK2 (> 50), NTRK1 (> 50), NTRK2 (> 50) Clean kinase profile, clean CEREP panel