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Assay Detail

CHEMBL5735192

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

Binding
Kinase Activity Inhibition Assay: Inhibition of ASK1 kinase activity was determined radiometrically using 33P substrate incorporation (Reaction Biology Corp., Malvern, Pa.). Briefly, recombinant human ASK1 protein, 20 μM substrate (myelin basic protein (MBP)), and compound (diluted from DMSO stock to give 1% final DMSO) were incubated in Base Reaction buffer (20 mM Hepes (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 0.02% Brij35, 0.02 mg/mL BSA, 0.1 mM Na3VO4, 2 mM DTT, 1% DMSO) at room temperature for 20 minutes. The reaction was initiated by addition of 10 μM 33P-ATP (specific activity 10 μCi/4l). The reaction was then allowed to proceed for 2 hours at room temperature. Upon reaction termination, the mixture was spotted on P81 exchange paper and kinase activity by incorporation of 33P into MBP substrate was detected by filter-binding method.
276
Total Activities
90
Compounds Tested
3
Activity Types
0
Assay Parameters

Assay Information

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

Target

Mitogen-activated protein kinase kinase kinase 5 (CHEMBL5285)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

ASK1 isoindolin-1-one inhibitors and methods of use thereof
(2020)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 92 8.22 9.52
kon 92 - -
k_off 92 - -

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL5787727 3 9.52
CHEMBL5946199 3 9.15
CHEMBL5967167 3 8.80
CHEMBL5795972 3 8.80
CHEMBL5841486 3 8.74
CHEMBL5746635 3 8.72
CHEMBL5985747 3 8.68
CHEMBL5879519 3 8.60
CHEMBL6060824 3 8.59
CHEMBL6057773 3 8.55
CHEMBL5775165 3 8.55
CHEMBL5865443 3 8.49
CHEMBL6022506 3 8.48
CHEMBL5862012 6 8.48
CHEMBL6019733 3 8.47
CHEMBL5886893 3 8.46
CHEMBL5895853 3 8.44
CHEMBL5771353 3 8.41
CHEMBL5782592 3 8.40
CHEMBL5819333 3 8.39
CHEMBL5868776 3 8.37
CHEMBL6058001 3 8.37
CHEMBL5914977 3 8.37
CHEMBL5804541 3 8.36
CHEMBL5750805 3 8.34
CHEMBL5908319 3 8.34
CHEMBL6049786 3 8.34
CHEMBL5876473 3 8.33
CHEMBL5906116 3 8.33
CHEMBL5939633 3 8.33

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL5787727 IC50 = 0.3 nM 9.52
CHEMBL5946199 IC50 = 0.7 nM 9.15
CHEMBL5795972 IC50 = 1.6 nM 8.80
CHEMBL5967167 IC50 = 1.6 nM 8.80
CHEMBL5841486 IC50 = 1.83 nM 8.74
CHEMBL5746635 IC50 = 1.9 nM 8.72
CHEMBL5985747 IC50 = 2.1 nM 8.68
CHEMBL5879519 IC50 = 2.5 nM 8.60
CHEMBL6060824 IC50 = 2.6 nM 8.59
CHEMBL5775165 IC50 = 2.8 nM 8.55
CHEMBL6057773 IC50 = 2.8 nM 8.55
CHEMBL5865443 IC50 = 3.2 nM 8.49
CHEMBL6022506 IC50 = 3.28 nM 8.48
CHEMBL5862012 IC50 = 3.3 nM 8.48
CHEMBL6019733 IC50 = 3.4 nM 8.47
CHEMBL5886893 IC50 = 3.5 nM 8.46
CHEMBL5895853 IC50 = 3.6 nM 8.44
CHEMBL5771353 IC50 = 3.9 nM 8.41
CHEMBL5782592 IC50 = 4.0 nM 8.40
CHEMBL5819333 IC50 = 4.1 nM 8.39
CHEMBL6058001 IC50 = 4.3 nM 8.37
CHEMBL5914977 IC50 = 4.3 nM 8.37
CHEMBL5868776 IC50 = 4.3 nM 8.37
CHEMBL5804541 IC50 = 4.4 nM 8.36
CHEMBL6049786 IC50 = 4.6 nM 8.34
CHEMBL5908319 IC50 = 4.6 nM 8.34
CHEMBL5750805 IC50 = 4.6 nM 8.34
CHEMBL5939633 IC50 = 4.7 nM 8.33
CHEMBL5876473 IC50 = 4.7 nM 8.33
CHEMBL5906116 IC50 = 4.7 nM 8.33
CHEMBL5857821 IC50 = 4.9 nM 8.31
CHEMBL5794733 IC50 = 4.9 nM 8.31
CHEMBL5758560 IC50 = 4.9 nM 8.31
CHEMBL6010208 IC50 = 4.9 nM 8.31
CHEMBL5787566 IC50 = 4.9 nM 8.31
CHEMBL6041520 IC50 = 5.0 nM 8.30
CHEMBL6029058 IC50 = 5.1 nM 8.29
CHEMBL5808077 IC50 = 5.1 nM 8.29
CHEMBL5987742 IC50 = 5.1 nM 8.29
CHEMBL5780163 IC50 = 5.2 nM 8.28
CHEMBL5765172 IC50 = 5.3 nM 8.28
CHEMBL5932507 IC50 = 5.2 nM 8.28
CHEMBL5963745 IC50 = 5.2 nM 8.28
CHEMBL5909721 IC50 = 5.2 nM 8.28
CHEMBL5792360 IC50 = 5.2 nM 8.28
CHEMBL6041862 IC50 = 5.3 nM 8.28
CHEMBL5880765 IC50 = 5.4 nM 8.27
CHEMBL5862012 IC50 = 5.4 nM 8.27
CHEMBL5976849 IC50 = 5.44 nM 8.26
CHEMBL6031940 IC50 = 5.52 nM 8.26