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Assay Detail

CHEMBL5733481

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

Binding
Human Recombinant Btk Enzyme Assay: To V-bottom 384-well plates were added test compounds, human recombinant Btk (1 nM, Invitrogen Corporation), fluoresceinated peptide (1.5 μM), ATP (20 μM), and assay buffer (20 mM HEPES pH 7.4, 10 mM MgCl2, 0.015% Brij 35 surfactant and 4 mM DTT in 1.6% DMSO), with a final volume of 30 μL. After incubating at room temperature for 60 min, the reaction was terminated by adding 45 μL of 35 mM EDTA to each sample. The reaction mixture was analyzed on the Caliper LABCHIP® 3000 (Caliper, Hopkinton, Mass.) by electrophoretic separation of the fluorescent substrate and phosphorylated product. Inhibition data were calculated by comparison to no enzyme control reactions for 100% inhibition and no inhibitor controls for 0% inhibition. Dose response curves were generated to determine the concentration required for inhibiting 50% of kinase activity (IC50). Compounds were dissolved at 10 mM in DMSO and evaluated at eleven concentrations.
600
Total Activities
167
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

Tyrosine-protein kinase BTK (CHEMBL5251)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Carbazole derivatives
(2019)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 200 9.18 10.34
kon 200 - -
k_off 200 - -

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL5909954 3 10.34
CHEMBL5789446 6 10.33
CHEMBL5749648 3 10.31
CHEMBL5858594 9 10.30
CHEMBL5799170 3 10.30
CHEMBL5772754 3 10.29
CHEMBL5848923 3 10.28
CHEMBL5983201 3 10.21
CHEMBL6042416 6 10.17
CHEMBL5819531 3 10.16
CHEMBL5862023 9 10.15
CHEMBL5808322 3 10.14
CHEMBL5958946 3 10.12
CHEMBL6053298 6 10.12
CHEMBL5989288 3 10.11
CHEMBL4240879 3 10.10
CHEMBL5935683 3 10.06
CHEMBL5965184 9 10.05
CHEMBL5935718 6 10.05
CHEMBL6057626 3 10.02
CHEMBL5863409 3 10.02
CHEMBL5801580 3 10.00
CHEMBL5958902 3 10.00
CHEMBL5891936 3 10.00
CHEMBL5942743 3 10.00
CHEMBL5847252 12 10.00
CHEMBL5948678 3 10.00
CHEMBL5793001 3 10.00
CHEMBL5931695 3 10.00
CHEMBL5884381 3 10.00

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL5909954 IC50 = 0.046 nM 10.34
CHEMBL5789446 IC50 = 0.047 nM 10.33
CHEMBL5749648 IC50 = 0.049 nM 10.31
CHEMBL5799170 IC50 = 0.05 nM 10.30
CHEMBL5858594 IC50 = 0.05 nM 10.30
CHEMBL5772754 IC50 = 0.051 nM 10.29
CHEMBL5848923 IC50 = 0.052 nM 10.28
CHEMBL5983201 IC50 = 0.061 nM 10.21
CHEMBL6042416 IC50 = 0.067 nM 10.17
CHEMBL5819531 IC50 = 0.069 nM 10.16
CHEMBL5862023 IC50 = 0.07 nM 10.15
CHEMBL6042416 IC50 = 0.07 nM 10.15
CHEMBL5808322 IC50 = 0.073 nM 10.14
CHEMBL5958946 IC50 = 0.076 nM 10.12
CHEMBL6053298 IC50 = 0.075 nM 10.12
CHEMBL5989288 IC50 = 0.078 nM 10.11
CHEMBL4240879 IC50 = 0.079 nM 10.10
CHEMBL5858594 IC50 = 0.087 nM 10.06
CHEMBL5935683 IC50 = 0.088 nM 10.06
CHEMBL5935718 IC50 = 0.09 nM 10.05
CHEMBL5965184 IC50 = 0.09 nM 10.05
CHEMBL6057626 IC50 = 0.095 nM 10.02
CHEMBL5863409 IC50 = 0.095 nM 10.02
CHEMBL5965184 IC50 = 0.1 nM 10.00
CHEMBL5884381 IC50 = 0.1 nM 10.00
CHEMBL5891936 IC50 = 0.1 nM 10.00
CHEMBL5958902 IC50 = 0.099 nM 10.00
CHEMBL5931695 IC50 = 0.1 nM 10.00
CHEMBL5942743 IC50 = 0.1 nM 10.00
CHEMBL5847252 IC50 = 0.1 nM 10.00
CHEMBL5793001 IC50 = 0.1 nM 10.00
CHEMBL5948678 IC50 = 0.1 nM 10.00
CHEMBL5801580 IC50 = 0.1 nM 10.00
CHEMBL6053298 IC50 = 0.1 nM 10.00
CHEMBL5755283 IC50 = 0.11 nM 9.96
CHEMBL5968758 IC50 = 0.11 nM 9.96
CHEMBL6054817 IC50 = 0.11 nM 9.96
CHEMBL5944790 IC50 = 0.12 nM 9.92
CHEMBL5948732 IC50 = 0.12 nM 9.92
CHEMBL5793668 IC50 = 0.12 nM 9.92
CHEMBL5988397 IC50 = 0.12 nM 9.92
CHEMBL5808911 IC50 = 0.13 nM 9.89
CHEMBL5789446 IC50 = 0.13 nM 9.89
CHEMBL5973174 IC50 = 0.14 nM 9.85
CHEMBL5959900 IC50 = 0.15 nM 9.82
CHEMBL6038830 IC50 = 0.16 nM 9.80
CHEMBL4248271 IC50 = 0.16 nM 9.80
CHEMBL5920793 IC50 = 0.16 nM 9.80
CHEMBL4248271 IC50 = 0.16 nM 9.80
CHEMBL4248719 IC50 = 0.17 nM 9.77