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Assay Detail

CHEMBL5736794

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Binding
ERK2 In vitro Inhibition Assay: Activity of ERK2 enzyme (Life Technologies) was determined using a time-resolved fluorescence format measuring the phosphorylation of a truncated version of Activating transcription factor 2 labelled with green fluorescent protein (ATF2-GFP) (Life Technologies). Assay reactions containing 50 mM Tris pH 7.5, 10 mM MgC2, 1 mM EGTA, 0.01% Triton X-100, 1 mM DTT, 2.5% DMSO, 0.4 μM ATF2-GFP, 20 μM ATP and 0.25 nM ERK2 were set up in the presence of compound and allowed to proceed for 30 min at room temperature. Reactions were then stopped using TR-FRET dilution buffer (Life Technologies), 25 mM EDTA and 2 nM Tb-Anti-pATF2 (Thr71) (Life Technologies). After a further incubation period of at least 30 minutes, fluorescence was read on a Pherastar reader (Lanthascreen optic module; excitation 340 nm, emission 520 nm (channel A), 495 nm (channel B)). The ratio between A and B counts was used to calculate signal. IC50 values were calculated using a sigmoidal dose response equation (Prism GraphPad software, La Jolla, Calif., USA).In the assays using ERK2, the compounds of Examples 125, 127, 162, 181, 183, 289, 299, 310 and 939 have IC50 values in the range from 1 μM to 10 μM, provide at least 50% inhibition of the activity at a concentration of 10 μM in the assay against ERK2 or would be expected to provide at least 50% inhibition of the activity at a concentration of 10 μM (based on the level of inhibition of the activity at concentrations of 3 μM) in the assay against ERK2.
2769
Total Activities
899
Compounds Tested
3
Activity Types
0
Assay Parameters

Assay Information

Assay Type Binding
Organism Homo sapiens
Confidence 9 — Direct single protein target
Curated By Autocuration

Target

Mitogen-activated protein kinase 1 (CHEMBL4040)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Benzolactam compounds as protein kinase inhibitors
(2021)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 923 7.84 8.96
kon 923 - -
k_off 923 - -

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL4871302 3 8.96
CHEMBL4849148 3 8.92
CHEMBL5952643 9 8.89
CHEMBL4847212 3 8.89
CHEMBL5930683 3 8.89
CHEMBL5883529 3 8.85
CHEMBL5875401 3 8.85
CHEMBL5776683 6 8.82
CHEMBL6019688 3 8.82
CHEMBL5812423 3 8.82
CHEMBL5886204 3 8.82
CHEMBL6034338 3 8.82
CHEMBL5822851 3 8.80
CHEMBL5923573 3 8.80
CHEMBL5746376 3 8.80
CHEMBL5760842 3 8.80
CHEMBL5774206 3 8.80
CHEMBL5860427 3 8.80
CHEMBL6046631 3 8.80
CHEMBL5979676 3 8.77
CHEMBL5965263 3 8.77
CHEMBL5905526 3 8.77
CHEMBL5918274 3 8.77
CHEMBL5903955 3 8.77
CHEMBL5857382 3 8.77
CHEMBL5788880 3 8.77
CHEMBL5803384 3 8.77
CHEMBL5788403 3 8.77
CHEMBL5900929 3 8.77
CHEMBL5823208 3 8.74

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL4871302 IC50 = 1.1 nM 8.96
CHEMBL4849148 IC50 = 1.2 nM 8.92
CHEMBL5952643 IC50 = 1.3 nM 8.89
CHEMBL4847212 IC50 = 1.3 nM 8.89
CHEMBL5930683 IC50 = 1.3 nM 8.89
CHEMBL5875401 IC50 = 1.4 nM 8.85
CHEMBL5883529 IC50 = 1.4 nM 8.85
CHEMBL5812423 IC50 = 1.5 nM 8.82
CHEMBL6019688 IC50 = 1.5 nM 8.82
CHEMBL5776683 IC50 = 1.5 nM 8.82
CHEMBL5886204 IC50 = 1.5 nM 8.82
CHEMBL6034338 IC50 = 1.5 nM 8.82
CHEMBL5822851 IC50 = 1.6 nM 8.80
CHEMBL5774206 IC50 = 1.6 nM 8.80
CHEMBL5760842 IC50 = 1.6 nM 8.80
CHEMBL5746376 IC50 = 1.6 nM 8.80
CHEMBL5860427 IC50 = 1.6 nM 8.80
CHEMBL6046631 IC50 = 1.6 nM 8.80
CHEMBL5923573 IC50 = 1.6 nM 8.80
CHEMBL5965263 IC50 = 1.7 nM 8.77
CHEMBL5788880 IC50 = 1.7 nM 8.77
CHEMBL5903955 IC50 = 1.7 nM 8.77
CHEMBL5803384 IC50 = 1.7 nM 8.77
CHEMBL5900929 IC50 = 1.7 nM 8.77
CHEMBL5776683 IC50 = 1.7 nM 8.77
CHEMBL5788403 IC50 = 1.7 nM 8.77
CHEMBL5905526 IC50 = 1.7 nM 8.77
CHEMBL5857382 IC50 = 1.7 nM 8.77
CHEMBL5979676 IC50 = 1.7 nM 8.77
CHEMBL5918274 IC50 = 1.7 nM 8.77
CHEMBL4857389 IC50 = 1.8 nM 8.74
CHEMBL5823208 IC50 = 1.8 nM 8.74
CHEMBL6064867 IC50 = 1.8 nM 8.74
CHEMBL5961403 IC50 = 1.8 nM 8.74
CHEMBL5768700 IC50 = 1.8 nM 8.74
CHEMBL5929439 IC50 = 1.8 nM 8.74
CHEMBL5933836 IC50 = 1.8 nM 8.74
CHEMBL5813710 IC50 = 1.9 nM 8.72
CHEMBL5929791 IC50 = 1.9 nM 8.72
CHEMBL4865318 IC50 = 1.9 nM 8.72
CHEMBL5754931 IC50 = 1.9 nM 8.72
CHEMBL5958179 IC50 = 1.9 nM 8.72
CHEMBL5860624 IC50 = 1.9 nM 8.72
CHEMBL6001381 IC50 = 1.9 nM 8.72
CHEMBL5907157 IC50 = 2.0 nM 8.70
CHEMBL5783614 IC50 = 2.0 nM 8.70
CHEMBL5888852 IC50 = 2.0 nM 8.70
CHEMBL5961659 IC50 = 2.0 nM 8.70
CHEMBL5999598 IC50 = 2.0 nM 8.70
CHEMBL5796891 IC50 = 2.0 nM 8.70