Donated Chemical Probes

Target	MAPK14
Targeted domain	Kinase domain (active site)
Mode of action	Inhibitor (type 2, ATP competitive)
Control	SR-321
Orthogonal probe	Skepinone-L FS-694
Recommended cellular usage concentration	< 100 nM
In vivo use	No
Donated by	SGC Frankfurt

Inhibitor/agonist potency: goal is < 100 nM (IC₅₀, K_d>/sub>)	Surpasses criterion: MAPK14: IC₅₀ =5 nM; MAPK11: IC₅₀ =32 nM (Radiometric activity assay ProQinase)
Selectivity within target family: > 30-fold	Surpasses criterion: Closest off-targets found in KinomeScan: DDR1, DDR2, KIT, MAP3K20, RPS6KA6, MYLK4 are > 200 fold selective in NanoBRET assay
Selectivity outside target family	Clean PDSP scan (45 targets)
On target cell activity for cell-based targets: goal is < 1 µM IC₅₀/EC₅₀	Surpasses criterion:NanaoBRET: MAPK14 IC₅₀ =3.7 nM; MAPK11 IC₅₀ = 10 nM (full-length protein in HEK cells); Western: inhibition of HSP27 (Ser82) < 1μM
On target cell activity for secreted targets: appropriate alternative such as mouse model or other mechanistic biological assay, e.g., explant culture	Inhibition of TNFα release in WB: IC₅₀ = 283 nM
Control compound (100 times less potent than the probe)	Surpasses criterion: SR-321: IC₅₀ MAPK14 = 13 µM and MAPK11 = 11 µM (NanoBRET), DDR2: IC₅₀ = 220 nM (NanoBRET), but no other kinase activity. Clean PDSP scan except for one off-target: TMEM97 Ki =2887.53 nM