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Assay Detail

CHEMBL3705207

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Binding
Enzymatic Assay: Compounds were tested in an enzymatic assay using human ERK-2 (Mitogen Activated Kinase 1), recombinantly expressed as an n-terminal 6-His fusion protein in E. coli and corresponding to aa 8-360. The substrate used was the fluorescent Omnia peptide S/T17 (Invitrogen of Carlsbad, Calif.; Cat. KNZ1171C). Test compounds were diluted in dimethylsulfoxide (DMSO) in 3-fold serial dilutions at 100× final concentrations. In addition to compound, the assay contained 50 mM HEPES [pH 7.3], 10 mM MgCl2, 1 mM DTT, 0.005% Triton-X100, 5 nM ERK-2 enzyme, 6.25 μM S/T17 peptide substrate and 25 μM ATP (corresponding to the observed Km) for a total reaction volume of 25 μL. The assay was run at ambient temperature in a white 384-well polypropylene plate (Nunc, Inc of Naperville, Ill.; Cat. 267462) collecting data every 50 seconds for approximately 30 minutes on an Envision plate reader (PerkinElmer, Inc. of Waltham, Mass.); Excitation 340 nm/Emission 495 nm.
42
Total Activities
42
Compounds Tested
1
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

Serine/threonine kinase inhibitors
(2014)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 42 8.12 9.10

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL3675723 1 9.10
CHEMBL3680608 1 8.80
CHEMBL3680610 1 8.66
CHEMBL3680609 1 8.62
CHEMBL3680618 1 8.59
CHEMBL3675720 1 8.54
CHEMBL3675718 1 8.54
CHEMBL3675706 1 8.52
CHEMBL3409576 1 8.52
CHEMBL3675719 1 8.51
CHEMBL3675701 1 8.49
CHEMBL3675711 1 8.48
CHEMBL3544964 RAVOXERTINIB 2.0 1 8.48
CHEMBL3675703 1 8.42
CHEMBL3409585 1 8.42
CHEMBL3409588 1 8.41
CHEMBL3675722 1 8.41
CHEMBL3675702 1 8.40
CHEMBL3680616 1 8.40
CHEMBL3675715 1 8.26
CHEMBL3675721 1 8.24
CHEMBL3409609 1 8.20
CHEMBL3675707 1 8.20
CHEMBL3680611 1 8.07
CHEMBL3675717 1 8.06
CHEMBL3680614 1 8.01
CHEMBL3675713 1 7.98
CHEMBL3675708 1 7.94
CHEMBL3680613 1 7.92
CHEMBL3675704 1 7.87

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL3675723 IC50 = 0.8 nM 9.10
CHEMBL3680608 IC50 = 1.6 nM 8.80
CHEMBL3680610 IC50 = 2.2 nM 8.66
CHEMBL3680609 IC50 = 2.4 nM 8.62
CHEMBL3680618 IC50 = 2.6 nM 8.59
CHEMBL3675720 IC50 = 2.9 nM 8.54
CHEMBL3675718 IC50 = 2.9 nM 8.54
CHEMBL3675706 IC50 = 3.0 nM 8.52
CHEMBL3409576 IC50 = 3.0 nM 8.52
CHEMBL3675719 IC50 = 3.1 nM 8.51
CHEMBL3675701 IC50 = 3.2 nM 8.49
CHEMBL3544964 RAVOXERTINIB IC50 = 3.3 nM 8.48
CHEMBL3675711 IC50 = 3.3 nM 8.48
CHEMBL3675703 IC50 = 3.8 nM 8.42
CHEMBL3409585 IC50 = 3.8 nM 8.42
CHEMBL3409588 IC50 = 3.9 nM 8.41
CHEMBL3675722 IC50 = 3.9 nM 8.41
CHEMBL3675702 IC50 = 4.0 nM 8.40
CHEMBL3680616 IC50 = 4.0 nM 8.40
CHEMBL3675715 IC50 = 5.5 nM 8.26
CHEMBL3675721 IC50 = 5.7 nM 8.24
CHEMBL3409609 IC50 = 6.3 nM 8.20
CHEMBL3675707 IC50 = 6.3 nM 8.20
CHEMBL3680611 IC50 = 8.5 nM 8.07
CHEMBL3675717 IC50 = 8.7 nM 8.06
CHEMBL3680614 IC50 = 9.8 nM 8.01
CHEMBL3675713 IC50 = 10.4 nM 7.98
CHEMBL3675708 IC50 = 11.5 nM 7.94
CHEMBL3680613 IC50 = 12.1 nM 7.92
CHEMBL3675704 IC50 = 13.6 nM 7.87
CHEMBL3675716 IC50 = 13.7 nM 7.86
CHEMBL3675709 IC50 = 13.9 nM 7.86
CHEMBL3680615 IC50 = 14.5 nM 7.84
CHEMBL3409594 IC50 = 15.6 nM 7.81
CHEMBL3675714 IC50 = 15.9 nM 7.80
CHEMBL3680612 IC50 = 15.8 nM 7.80
CHEMBL3409586 IC50 = 35.7 nM 7.45
CHEMBL3675712 IC50 = 39.5 nM 7.40
CHEMBL3409598 IC50 = 51.8 nM 7.29
CHEMBL3675710 IC50 = 53.0 nM 7.28
CHEMBL3675705 IC50 = 145.6 nM 6.84
CHEMBL3680617 IC50 = 249.1 nM 6.60