Assay Detail

CHEMBL3887900

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Binding

ERK2 (20 pM) Kinase Assay: Compound potency against activated ERK2 was determined using a kinase assay that measures ERK2-catalyzed phosphorylation of biotinylated ERKtide peptide substrate ([Biotin]-AHA-K-R-E-L-V-E-P-L-T-P-S-G-E-A-P-N-Q-A-L-L-R-[NH2], the peptide sequence derived from EGF receptor: SEQ ID NO:1). The assay was carried out in 20 mM HEPES [pH 7.5], 5 mM MgCl2, 1 mM DTT, 0.01% Tween-20, 0.05% BSA using 0.02 nM ERK2, 400 nM ERKtide peptide and 35 uM ATP (all concentrations are final in the reaction) in a total volume of 10.25 uL. A 16-point, half-log dilution series of compounds at 41x final concentration was used for generating IC50 curves. Compound dilution series were prepared in 100% DMSO. ERK2 was preincubated with compounds for 30 minutes at ambient temperature. Reaction was initiated by addition of a substrate cocktail of ERKtide peptide and ATP and was allowed to proceed for 4 hours at ambient temperature. Reaction was terminated by addition of 10 uL of a 2x stop buffer consisting of 100 mM Tris-Cl [pH 7.5], 25 mM EDTA, 0.01% Tween 20, 20 ug/mL of AlphaScreen Protein A Acceptor Beads, 20 ug/mL of Streptavidin Donor Beads (PerkinElmer, Waltham, Mass.), and 1:1000 dilution phospho-EGF Receptor (Thr669) antibody (Cat #8808, Cell Signaling Technology, Danvers, Mass.). Terminated reactions were read, after overnight incubation in the dark, on an EnVision Multilabel Plate Reader (PerkinElmer, Waltham, Mass.), with excitation and emission wavelengths set to 680 nm and 570 nm, respectively. IC50 values were determined using a four-parameter fit.

ChEMBL ↗

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Target

Mitogen-activated protein kinase 1 (CHEMBL4040) ↗

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Primary target Mitogen-activated protein kinase 1

CHEMBL4040

Anchored from Assay target.

Protein identity Mitogen-activated protein kinase 1

Disease frame Disease evidence pending

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280

Total Activities

261

Compounds Tested

Activity Types

Assay Parameters

Assay Information

Assay Type	Binding
Organism	Homo sapiens
Confidence	8 — Homologous single protein target
Curated By	Autocuration

Target

Mitogen-activated protein kinase 1 (CHEMBL4040) ↗

Type	SINGLE PROTEIN
Organism	Homo sapiens

Publication

Aminoheteroaryl benzamides as kinase inhibitors

(2016)

Activity Statistics

Type	Count	Avg pChEMBL	Best pChEMBL
IC50	280	9.57	11.00

Compounds Tested

Compound	Name	Phase	Activities	Best pChEMBL
CHEMBL4108345	—	—	1	11.00
CHEMBL4114147	—	—	1	11.00
CHEMBL4109252	—	—	1	10.99
CHEMBL3957670	—	—	1	10.98
CHEMBL4107945	—	—	1	10.97
CHEMBL4109456	—	—	1	10.96
CHEMBL3923033	—	—	1	10.92
CHEMBL4110508	—	—	1	10.92
CHEMBL4108076	—	—	1	10.90
CHEMBL3956128	—	—	1	10.86
CHEMBL4107980	—	—	1	10.86
CHEMBL4109206	—	—	1	10.85
CHEMBL4115307	—	—	1	10.84
CHEMBL3923915	—	—	1	10.84
CHEMBL4109955	—	—	1	10.82
CHEMBL4106909	—	—	1	10.80
CHEMBL4113157	—	—	1	10.77
CHEMBL4107833	—	—	1	10.76
CHEMBL4114947	—	—	1	10.76
CHEMBL3913471	—	—	1	10.75
CHEMBL4108962	—	—	1	10.74
CHEMBL3905175	—	—	2	10.73
CHEMBL4114300	—	—	1	10.73
CHEMBL4106814	—	—	1	10.71
CHEMBL3930522	—	—	1	10.71
CHEMBL4113452	—	—	1	10.70
CHEMBL4114593	—	—	1	10.70
CHEMBL4108926	—	—	1	10.70
CHEMBL4108313	—	—	1	10.70
CHEMBL4115371	—	—	1	10.69

Activity Data

Compound	Name	Type	Rel.	Value	Units	pChEMBL
CHEMBL4108345	—	IC50	=	0.01	nM	11.00
CHEMBL4114147	—	IC50	=	0.01	nM	11.00
CHEMBL4109252	—	IC50	=	0.0103	nM	10.99
CHEMBL3957670	—	IC50	=	0.0104	nM	10.98
CHEMBL4107945	—	IC50	=	0.0107	nM	10.97
CHEMBL4109456	—	IC50	=	0.011	nM	10.96
CHEMBL4110508	—	IC50	=	0.0121	nM	10.92
CHEMBL3923033	—	IC50	=	0.0119	nM	10.92
CHEMBL4108076	—	IC50	=	0.0127	nM	10.90
CHEMBL4107980	—	IC50	=	0.0139	nM	10.86
CHEMBL3956128	—	IC50	=	0.0139	nM	10.86
CHEMBL4109206	—	IC50	=	0.014	nM	10.85
CHEMBL3923915	—	IC50	=	0.0143	nM	10.84
CHEMBL4115307	—	IC50	=	0.0143	nM	10.84
CHEMBL4109955	—	IC50	=	0.0153	nM	10.82
CHEMBL4106909	—	IC50	=	0.0158	nM	10.80
CHEMBL4113157	—	IC50	=	0.0168	nM	10.77
CHEMBL4107833	—	IC50	=	0.0173	nM	10.76
CHEMBL4114947	—	IC50	=	0.0175	nM	10.76
CHEMBL3913471	—	IC50	=	0.0176	nM	10.75
CHEMBL4108962	—	IC50	=	0.0183	nM	10.74
CHEMBL4114300	—	IC50	=	0.0187	nM	10.73
CHEMBL3905175	—	IC50	=	0.0187	nM	10.73
CHEMBL4106814	—	IC50	=	0.0197	nM	10.71
CHEMBL3930522	—	IC50	=	0.0194	nM	10.71
CHEMBL4113452	—	IC50	=	0.0199	nM	10.70
CHEMBL4114593	—	IC50	=	0.0198	nM	10.70
CHEMBL4108313	—	IC50	=	0.02	nM	10.70
CHEMBL4108926	—	IC50	=	0.0201	nM	10.70
CHEMBL4115371	—	IC50	=	0.0205	nM	10.69
CHEMBL4108747	—	IC50	=	0.0205	nM	10.69
CHEMBL4113751	—	IC50	=	0.0208	nM	10.68
CHEMBL4107676	—	IC50	=	0.0209	nM	10.68
CHEMBL4113025	—	IC50	=	0.0208	nM	10.68
CHEMBL3928081	—	IC50	=	0.021	nM	10.68
CHEMBL4112962	—	IC50	=	0.0209	nM	10.68
CHEMBL4110468	—	IC50	=	0.0208	nM	10.68
CHEMBL3924033	—	IC50	=	0.0217	nM	10.66
CHEMBL4112079	—	IC50	=	0.0222	nM	10.65
CHEMBL3924033	—	IC50	=	0.0225	nM	10.65
CHEMBL3891114	—	IC50	=	0.0229	nM	10.64
CHEMBL4108968	—	IC50	=	0.023	nM	10.64
CHEMBL3905175	—	IC50	=	0.0233	nM	10.63
CHEMBL3924033	—	IC50	=	0.0234	nM	10.63
CHEMBL4108246	—	IC50	=	0.0242	nM	10.62
CHEMBL3924033	—	IC50	=	0.0239	nM	10.62
CHEMBL4113613	—	IC50	=	0.0248	nM	10.61
CHEMBL4106578	—	IC50	=	0.0245	nM	10.61
CHEMBL4106790	—	IC50	=	0.0252	nM	10.60
CHEMBL4114268	—	IC50	=	0.0258	nM	10.59