Assay Detail
Binding
CHEMBL5735964
Review assay metadata, readout intent, target linkage, and publication context from the same page.
LanthaScreen Assay: BTK: TR-FRET LanthaScreen assays were performed by incubating a dilution series of inhibitor concentrations with 50 μM ATP, 100 nM FAM-Srctide peptide substrate (5FAM-GEEPLYWSFPAKKK-NH2, SEQ ID NO: 1, Molecular Devices, RP7595) and 70 pM of human full-length BTK Kinase (expressed in Sf9 insect cells and purified in-house). The assays were performed with and without pre-incubating the inhibitors with the enzyme for 60 minutes before starting the kinase reaction by adding ATP and the peptide substrate. Samples containing enzyme but no inhibitor were included to determine the maximal extent of reaction. Samples containing no enzyme served as the negative control. The kinase reaction mixtures were incubated at room temperature for 60 minutes before stopping the kinase activity by the addition of 15 mM EDTA. The extent of peptide phosphorylation by BTK was detected using a Terbium-conjugated anti-phospho-Tyrosine antibody (Tb-PT66 antibody, Invitrogen # PV3557). Phosphorylation of peptide substrate was measured by determining the ratio of 520/495 nm on an Envision Multi-label Reader (Perkin Elmer) and IC50 values were calculated by fitting the data to a four-parameter equation using XLFit4 (IDBS).
498
Total Activities
140
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 |
Publication
Bruton's tyrosine kinase inhibitors
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| IC50 | 166 | 8.24 | 9.77 |
| kon | 166 | - | - |
| k_off | 166 | - | - |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL5781756 | — | — | 3 | 9.77 |
| CHEMBL4447328 | — | — | 3 | 9.74 |
| CHEMBL5752119 | — | — | 6 | 9.60 |
| CHEMBL5842396 | — | — | 9 | 9.43 |
| CHEMBL5916210 | — | — | 3 | 9.42 |
| CHEMBL5842964 | — | — | 3 | 9.40 |
| CHEMBL5882418 | — | — | 3 | 9.40 |
| CHEMBL5936062 | — | — | 3 | 9.39 |
| CHEMBL5742930 | — | — | 9 | 9.37 |
| CHEMBL5916403 | — | — | 3 | 9.36 |
| CHEMBL5808447 | — | — | 6 | 9.34 |
| CHEMBL5926524 | — | — | 3 | 9.32 |
| CHEMBL5826277 | — | — | 3 | 9.27 |
| CHEMBL5927942 | — | — | 3 | 9.22 |
| CHEMBL4456283 | — | — | 3 | 9.19 |
| CHEMBL5895064 | — | — | 3 | 9.15 |
| CHEMBL5868553 | — | — | 3 | 9.13 |
| CHEMBL5912536 | — | — | 3 | 9.11 |
| CHEMBL5820322 | — | — | 3 | 9.10 |
| CHEMBL5939859 | — | — | 3 | 9.09 |
| CHEMBL5838675 | — | — | 9 | 9.09 |
| CHEMBL5941034 | — | — | 3 | 9.00 |
| CHEMBL5752464 | — | — | 9 | 9.00 |
| CHEMBL5948246 | — | — | 3 | 9.00 |
| CHEMBL5960732 | — | — | 3 | 8.96 |
| CHEMBL6063311 | — | — | 3 | 8.96 |
| CHEMBL5790269 | — | — | 3 | 8.92 |
| CHEMBL5775965 | — | — | 3 | 8.92 |
| CHEMBL5954821 | — | — | 3 | 8.92 |
| CHEMBL5774120 | — | — | 3 | 8.92 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL5781756 | — | IC50 | = | 0.17 | nM | 9.77 |
| CHEMBL4447328 | — | IC50 | = | 0.18 | nM | 9.74 |
| CHEMBL5752119 | — | IC50 | = | 0.25 | nM | 9.60 |
| CHEMBL5842396 | — | IC50 | = | 0.37 | nM | 9.43 |
| CHEMBL5916210 | — | IC50 | = | 0.38 | nM | 9.42 |
| CHEMBL5842964 | — | IC50 | = | 0.4 | nM | 9.40 |
| CHEMBL5882418 | — | IC50 | = | 0.4 | nM | 9.40 |
| CHEMBL5936062 | — | IC50 | = | 0.41 | nM | 9.39 |
| CHEMBL5742930 | — | IC50 | = | 0.43 | nM | 9.37 |
| CHEMBL5916403 | — | IC50 | = | 0.44 | nM | 9.36 |
| CHEMBL5808447 | — | IC50 | = | 0.46 | nM | 9.34 |
| CHEMBL5926524 | — | IC50 | = | 0.48 | nM | 9.32 |
| CHEMBL5826277 | — | IC50 | = | 0.54 | nM | 9.27 |
| CHEMBL5927942 | — | IC50 | = | 0.6 | nM | 9.22 |
| CHEMBL4456283 | — | IC50 | = | 0.64 | nM | 9.19 |
| CHEMBL5895064 | — | IC50 | = | 0.7 | nM | 9.15 |
| CHEMBL5868553 | — | IC50 | = | 0.74 | nM | 9.13 |
| CHEMBL5912536 | — | IC50 | = | 0.77 | nM | 9.11 |
| CHEMBL5820322 | — | IC50 | = | 0.79 | nM | 9.10 |
| CHEMBL5939859 | — | IC50 | = | 0.81 | nM | 9.09 |
| CHEMBL5838675 | — | IC50 | = | 0.81 | nM | 9.09 |
| CHEMBL5838675 | — | IC50 | = | 1.0 | nM | 9.00 |
| CHEMBL5941034 | — | IC50 | = | 1.0 | nM | 9.00 |
| CHEMBL5742930 | — | IC50 | = | 1.0 | nM | 9.00 |
| CHEMBL5752464 | — | IC50 | = | 1.0 | nM | 9.00 |
| CHEMBL5948246 | — | IC50 | = | 1.0 | nM | 9.00 |
| CHEMBL6063311 | — | IC50 | = | 1.1 | nM | 8.96 |
| CHEMBL5960732 | — | IC50 | = | 1.1 | nM | 8.96 |
| CHEMBL5790269 | — | IC50 | = | 1.2 | nM | 8.92 |
| CHEMBL5775965 | — | IC50 | = | 1.2 | nM | 8.92 |
| CHEMBL5774120 | — | IC50 | = | 1.2 | nM | 8.92 |
| CHEMBL5954821 | — | IC50 | = | 1.2 | nM | 8.92 |
| CHEMBL6048351 | — | IC50 | = | 1.3 | nM | 8.89 |
| CHEMBL5903044 | — | IC50 | = | 1.3 | nM | 8.89 |
| CHEMBL5885541 | — | IC50 | = | 1.28 | nM | 8.89 |
| CHEMBL5949878 | — | IC50 | = | 1.3 | nM | 8.89 |
| CHEMBL5787151 | — | IC50 | = | 1.3 | nM | 8.89 |
| CHEMBL5853497 | — | IC50 | = | 1.3 | nM | 8.89 |
| CHEMBL5966610 | — | IC50 | = | 1.3 | nM | 8.89 |
| CHEMBL6047250 | — | IC50 | = | 1.3 | nM | 8.89 |
| CHEMBL5842396 | — | IC50 | = | 1.4 | nM | 8.85 |
| CHEMBL5835317 | — | IC50 | = | 1.4 | nM | 8.85 |
| CHEMBL5887361 | — | IC50 | = | 1.4 | nM | 8.85 |
| CHEMBL5963260 | — | IC50 | = | 1.5 | nM | 8.82 |
| CHEMBL5795141 | — | IC50 | = | 1.5 | nM | 8.82 |
| CHEMBL5798403 | — | IC50 | = | 1.6 | nM | 8.80 |
| CHEMBL5784044 | — | IC50 | = | 1.7 | nM | 8.77 |
| CHEMBL5952459 | — | IC50 | = | 1.7 | nM | 8.77 |
| CHEMBL5841573 | — | IC50 | = | 1.8 | nM | 8.74 |
| CHEMBL5815887 | — | IC50 | = | 1.8 | nM | 8.74 |