Assay Detail
Binding
CHEMBL5736237
Review assay metadata, readout intent, target linkage, and publication context from the same page.
BTK Assay: An HTRF assay (Cisbio KinEASE-TK cat #62TK0PEC) was performed to quantitate the ability of test compounds to inhibit BTK mediated phosphorylation of substrate. Assays were assembled in 384 well plates where 6 nM of full-length human His-tagged BTK (Life Technologies cat # PV3587) and test compound at varying concentrations were preincubated for 15 minutes at 28° C. Then, 1 uM of TK substrate-biotin and 30 uM ATP were added and incubated for an additional 30 minutes at 28° C. Phospohrylation was detected by adding 62.5 nM Streptavidin-XL665 and TK-Antibody Cryptate diluted 1:100 in HTRF detection buffer (Cisbio cat #62SDBRDF) and incubated for 60 minutes at RT. The plate was read on an Envision plate reader and the fluorescence is measured at 620 nm (cryptate) and 665 nm (XL665). A ratio is calculated (665/620) and converted to POC relative to control and blank wells.Assay Buffer:50 mM HEPES (Invitrogen #15630), 0.01% Brij-35 (sigma # B4184), 10 mM MgCl2 (Sigma M1028), 1 mM EGTA (Ambion AM9262) and 100 uM sodium orthovanedate (Sigma S6508), 1 mM DTT (Sigma D5545) and 10 nM supplement enzyme buffer (Cisbio cat #61SEBALB).
468
Total Activities
151
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
Heteroaromatic compounds as BTK inhibitors
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| IC50 | 156 | 7.89 | 9.70 |
| kon | 156 | - | - |
| k_off | 156 | - | - |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL5864486 | — | — | 3 | 9.70 |
| CHEMBL5834993 | — | — | 6 | 9.30 |
| CHEMBL5782411 | — | — | 3 | 9.30 |
| CHEMBL5810745 | — | — | 3 | 9.22 |
| CHEMBL5989330 | — | — | 3 | 9.15 |
| CHEMBL6016166 | — | — | 3 | 9.15 |
| CHEMBL5865771 | — | — | 3 | 9.14 |
| CHEMBL4447328 | — | — | 3 | 9.14 |
| CHEMBL5890539 | — | — | 3 | 9.11 |
| CHEMBL5847425 | — | — | 3 | 9.10 |
| CHEMBL5852587 | — | — | 3 | 9.10 |
| CHEMBL5852514 | — | — | 3 | 9.10 |
| CHEMBL5937691 | — | — | 3 | 9.10 |
| CHEMBL5981317 | — | — | 3 | 9.10 |
| CHEMBL5944649 | — | — | 3 | 9.06 |
| CHEMBL6011689 | — | — | 3 | 9.05 |
| CHEMBL5759776 | — | — | 3 | 9.05 |
| CHEMBL5893757 | — | — | 3 | 9.05 |
| CHEMBL6000192 | — | — | 3 | 9.05 |
| CHEMBL5833723 | — | — | 3 | 9.03 |
| CHEMBL5921140 | — | — | 3 | 9.00 |
| CHEMBL5921734 | — | — | 3 | 8.96 |
| CHEMBL5989799 | — | — | 6 | 8.96 |
| CHEMBL5918461 | — | — | 3 | 8.96 |
| CHEMBL5920832 | — | — | 3 | 8.92 |
| CHEMBL5880319 | — | — | 3 | 8.92 |
| CHEMBL5984238 | — | — | 3 | 8.85 |
| CHEMBL5744138 | — | — | 3 | 8.82 |
| CHEMBL5886894 | — | — | 3 | 8.80 |
| CHEMBL5843036 | — | — | 3 | 8.77 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL5864486 | — | IC50 | = | 0.2 | nM | 9.70 |
| CHEMBL5834993 | — | IC50 | = | 0.5 | nM | 9.30 |
| CHEMBL5782411 | — | IC50 | = | 0.5 | nM | 9.30 |
| CHEMBL5810745 | — | IC50 | = | 0.6 | nM | 9.22 |
| CHEMBL6016166 | — | IC50 | = | 0.7 | nM | 9.15 |
| CHEMBL5989330 | — | IC50 | = | 0.7 | nM | 9.15 |
| CHEMBL5865771 | — | IC50 | = | 0.73 | nM | 9.14 |
| CHEMBL4447328 | — | IC50 | = | 0.73 | nM | 9.14 |
| CHEMBL5890539 | — | IC50 | = | 0.77 | nM | 9.11 |
| CHEMBL5852514 | — | IC50 | = | 0.8 | nM | 9.10 |
| CHEMBL5852587 | — | IC50 | = | 0.79 | nM | 9.10 |
| CHEMBL5847425 | — | IC50 | = | 0.79 | nM | 9.10 |
| CHEMBL5937691 | — | IC50 | = | 0.8 | nM | 9.10 |
| CHEMBL5981317 | — | IC50 | = | 0.8 | nM | 9.10 |
| CHEMBL5944649 | — | IC50 | = | 0.87 | nM | 9.06 |
| CHEMBL6000192 | — | IC50 | = | 0.9 | nM | 9.05 |
| CHEMBL5759776 | — | IC50 | = | 0.9 | nM | 9.05 |
| CHEMBL5893757 | — | IC50 | = | 0.9 | nM | 9.05 |
| CHEMBL6011689 | — | IC50 | = | 0.89 | nM | 9.05 |
| CHEMBL5833723 | — | IC50 | = | 0.93 | nM | 9.03 |
| CHEMBL5921140 | — | IC50 | = | 1.0 | nM | 9.00 |
| CHEMBL5989799 | — | IC50 | = | 1.1 | nM | 8.96 |
| CHEMBL5921734 | — | IC50 | = | 1.1 | nM | 8.96 |
| CHEMBL5918461 | — | IC50 | = | 1.1 | nM | 8.96 |
| CHEMBL5920832 | — | IC50 | = | 1.2 | nM | 8.92 |
| CHEMBL5880319 | — | IC50 | = | 1.2 | nM | 8.92 |
| CHEMBL5984238 | — | IC50 | = | 1.4 | nM | 8.85 |
| CHEMBL5744138 | — | IC50 | = | 1.5 | nM | 8.82 |
| CHEMBL5886894 | — | IC50 | = | 1.6 | nM | 8.80 |
| CHEMBL5843036 | — | IC50 | = | 1.7 | nM | 8.77 |
| CHEMBL5874890 | — | IC50 | = | 1.9 | nM | 8.72 |
| CHEMBL5890655 | — | IC50 | = | 1.9 | nM | 8.72 |
| CHEMBL6064610 | — | IC50 | = | 2.0 | nM | 8.70 |
| CHEMBL5861890 | — | IC50 | = | 2.0 | nM | 8.70 |
| CHEMBL5840071 | — | IC50 | = | 2.1 | nM | 8.68 |
| CHEMBL5854298 | — | IC50 | = | 2.1 | nM | 8.68 |
| CHEMBL5983452 | — | IC50 | = | 2.2 | nM | 8.66 |
| CHEMBL5898333 | — | IC50 | = | 2.3 | nM | 8.64 |
| CHEMBL6008438 | — | IC50 | = | 2.7 | nM | 8.57 |
| CHEMBL5867921 | — | IC50 | = | 2.8 | nM | 8.55 |
| CHEMBL6051455 | — | IC50 | = | 3.0 | nM | 8.52 |
| CHEMBL5776066 | — | IC50 | = | 3.0 | nM | 8.52 |
| CHEMBL5830755 | — | IC50 | = | 3.2 | nM | 8.49 |
| CHEMBL5966328 | — | IC50 | = | 3.2 | nM | 8.49 |
| CHEMBL4456912 | — | IC50 | = | 3.2 | nM | 8.49 |
| CHEMBL5767485 | — | IC50 | = | 3.3 | nM | 8.48 |
| CHEMBL5866577 | — | IC50 | = | 3.4 | nM | 8.47 |
| CHEMBL5973202 | — | IC50 | = | 3.5 | nM | 8.46 |
| CHEMBL5891776 | — | IC50 | = | 3.5 | nM | 8.46 |
| CHEMBL5989799 | — | IC50 | = | 3.5 | nM | 8.46 |