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Assay Detail

CHEMBL5732253

Review assay metadata, readout intent, target linkage, and publication context from the same page.

Binding
Radioactive Binding Assay: The radioactive filter binding assay is standardized using recombinant human activated BRAF (V599E) kinase (Cat No. 14-557) and kinase dead MEK1 (K97R) (Cat No. 14-737) procured from Upstate. The incorporation of 32P into MEK1 (K97R) by BRAF (V599E) is measured with final assay buffer conditions of 50 mM Tris pH 7.5, 10 mM MgCl2, 1 mM DTT, 100 mM sucrose, 100 μM sodium orthovanadate, 5 μM ATP and 2 μCi [γ 32P] ATP and 500 mg MEK1 Kinase dead substrate. The enzymatic reaction is stopped after 120 minutes with 8N HCl (hydrochloric acid) and 1 mM ATP. The solution is spotted on P81 filter paper and washed 4 times with 0.75% orthophosphoric acid and lastly with acetone. The dried P81 filter papers are read in a Micro-beta Trilux scintillation counter. The final concentration of DMSO is 1% in the assay. Compounds are screened at 10 μM concentration with pre-incubation of the enzymes in the presence of test compound for 45 minutes. Compounds of the invention were found to be inactive in this assy, e.g. ex. 33 (13% inhibition at 10 uM), ex. 1A (0% inhibition at 10 uM).
303
Total Activities
100
Compounds Tested
3
Activity Types
0
Assay Parameters

Assay Information

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

Publication

Compounds and compositions as inhibitors of MEK
(2017)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 101 7.58 9.00
kon 101 - -
k_off 101 - -

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL3946736 3 9.00
CHEMBL3980017 3 8.82
CHEMBL3930250 3 8.77
CHEMBL3943311 3 8.70
CHEMBL3916718 3 8.62
CHEMBL3974369 3 8.59
CHEMBL3955338 3 8.59
CHEMBL3971701 3 8.57
CHEMBL3934047 3 8.57
CHEMBL3918610 3 8.52
CHEMBL5970584 3 8.52
CHEMBL3936697 3 8.44
CHEMBL5939759 3 8.43
CHEMBL3961528 3 8.43
CHEMBL3934040 3 8.42
CHEMBL3967019 3 8.37
CHEMBL3894284 3 8.37
CHEMBL5882359 3 8.35
CHEMBL3913667 3 8.34
CHEMBL3951042 3 8.33
CHEMBL3904439 3 8.30
CHEMBL3963847 3 8.28
CHEMBL5744841 3 8.24
CHEMBL3913739 6 8.21
CHEMBL5866714 3 8.15
CHEMBL3936680 3 8.15
CHEMBL3960631 3 8.14
CHEMBL3904177 3 8.07
CHEMBL3940408 3 8.07
CHEMBL5980351 3 8.05

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL3946736 IC50 = 1.0 nM 9.00
CHEMBL3980017 IC50 = 1.5 nM 8.82
CHEMBL3930250 IC50 = 1.7 nM 8.77
CHEMBL3943311 IC50 = 2.0 nM 8.70
CHEMBL3916718 IC50 = 2.4 nM 8.62
CHEMBL3955338 IC50 = 2.6 nM 8.59
CHEMBL3974369 IC50 = 2.6 nM 8.59
CHEMBL3971701 IC50 = 2.7 nM 8.57
CHEMBL3934047 IC50 = 2.7 nM 8.57
CHEMBL5970584 IC50 = 3.0 nM 8.52
CHEMBL3918610 IC50 = 3.0 nM 8.52
CHEMBL3936697 IC50 = 3.6 nM 8.44
CHEMBL5939759 IC50 = 3.7 nM 8.43
CHEMBL3961528 IC50 = 3.7 nM 8.43
CHEMBL3934040 IC50 = 3.8 nM 8.42
CHEMBL3894284 IC50 = 4.3 nM 8.37
CHEMBL3967019 IC50 = 4.3 nM 8.37
CHEMBL5882359 IC50 = 4.5 nM 8.35
CHEMBL3913667 IC50 = 4.6 nM 8.34
CHEMBL3951042 IC50 = 4.7 nM 8.33
CHEMBL3904439 IC50 = 5.0 nM 8.30
CHEMBL3963847 IC50 = 5.3 nM 8.28
CHEMBL5744841 IC50 = 5.7 nM 8.24
CHEMBL3913739 IC50 = 6.2 nM 8.21
CHEMBL3913739 IC50 = 6.2 nM 8.21
CHEMBL3936680 IC50 = 7.1 nM 8.15
CHEMBL5866714 IC50 = 7.0 nM 8.15
CHEMBL3960631 IC50 = 7.3 nM 8.14
CHEMBL3940408 IC50 = 8.5 nM 8.07
CHEMBL3904177 IC50 = 8.5 nM 8.07
CHEMBL5980351 IC50 = 9.0 nM 8.05
CHEMBL3914635 IC50 = 9.2 nM 8.04
CHEMBL3941176 IC50 = 10.0 nM 8.00
CHEMBL3908754 IC50 = 10.0 nM 8.00
CHEMBL3933193 IC50 = 10.0 nM 8.00
CHEMBL3936824 IC50 = 11.0 nM 7.96
CHEMBL3918375 IC50 = 11.0 nM 7.96
CHEMBL3922238 IC50 = 11.0 nM 7.96
CHEMBL3923663 IC50 = 11.0 nM 7.96
CHEMBL3951376 IC50 = 11.0 nM 7.96
CHEMBL5790979 IC50 = 12.0 nM 7.92
CHEMBL3946095 IC50 = 13.0 nM 7.89
CHEMBL3957047 IC50 = 13.0 nM 7.89
CHEMBL3927711 IC50 = 15.0 nM 7.82
CHEMBL3966473 IC50 = 16.0 nM 7.80
CHEMBL3889657 IC50 = 18.0 nM 7.75
CHEMBL3946632 IC50 = 18.0 nM 7.75
CHEMBL5741764 IC50 = 19.0 nM 7.72
CHEMBL3935342 IC50 = 19.0 nM 7.72
CHEMBL3906048 IC50 = 19.0 nM 7.72