Assay Detail
Binding
CHEMBL5737840
Review assay metadata, readout intent, target linkage, and publication context from the same page.
Enzyme Inhibition Assay: Enzyme inhibition studies were performed using recombinant JAK1 (amino acids 866-1154, Life Technologies, #PV4774, Carlsbad, Calif.), JAK2 (amino acids 831-1132), or JAK3 (amino acids 781-1124) under buffer conditions of 50 mM HEPES pH 7.3, 1 mM DTT, 0.01% Tween® 20, 50 μg/mL BSA, and 10 mM MgCl2. JAK enzyme was expressed as an N-terminal GST fusion in insect cells and purified by glutathione-affinity and size-exclusion chromatographies. Enzymes were assayed both at their respective ATP Km (JAK1: 55 μM, JAK2: 15 μM, JAK3: 3 μM) and the approximated high end of physiological ATP concentration of 5 mM, in the presence of inhibitor dosed at 30, 3, 0.3, 0.03, 0.003 and 0 μM final test concentrations. For JAK1, 6 nM of enzyme (for Km ATP assay) or 4 nM enzyme (for high ATP assay) was incubated with 1.5 μM peptide substrate (FITC-C6-KKHTDDGYMPMSPGVA-NH2 (SEQ ID NO:1), Intonation, Boston, Mass.). For JAK2, 0.8 nM of enzyme (for Km ATP assay) or 0.3 nM enzyme (for high ATP assay) was incubated with 1.5 μM peptide substrate (5FAM-GEEPLYWSFPAKKK-NH2 (SEQ ID NO:2), Intonation, Boston, Mass.). For JAK3, 0.2 nM of enzyme (for Km ATP assay) or 0.1 nM enzyme (for high ATP assay) was incubated with 1.5 μM peptide substrate (5FAM-GEEPLYWSFPAKKK-NH2 (SEQ ID NO:2), Intonation, Boston, Mass.). Phosphorylated and unphosphorylated peptides were separated and quantified by a Caliper LC3000 system (Caliper Life Sciences, MA) for calculating percent inhibition.
207
Total Activities
69
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 methods for inhibiting JAK
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| IC50 | 69 | 5.15 | 6.49 |
| kon | 69 | - | - |
| k_off | 69 | - | - |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL4060528 | — | — | 3 | 6.49 |
| CHEMBL5890455 | — | — | 3 | 6.10 |
| CHEMBL5923623 | — | — | 3 | 5.82 |
| CHEMBL4577393 | — | — | 3 | 5.80 |
| CHEMBL4085905 | — | — | 3 | 5.75 |
| CHEMBL4064366 | — | — | 3 | 5.75 |
| CHEMBL6045935 | — | — | 3 | 5.71 |
| CHEMBL5995592 | — | — | 3 | 5.69 |
| CHEMBL5912136 | — | — | 3 | 5.60 |
| CHEMBL5756048 | — | — | 3 | 5.59 |
| CHEMBL5989133 | — | — | 3 | 5.50 |
| CHEMBL5748427 | — | — | 3 | 5.38 |
| CHEMBL4092807 | — | — | 3 | 5.30 |
| CHEMBL5755930 | — | — | 3 | 5.29 |
| CHEMBL4450982 | — | — | 3 | 5.27 |
| CHEMBL5953308 | — | — | 3 | 5.21 |
| CHEMBL5830364 | — | — | 3 | 5.19 |
| CHEMBL5785335 | — | — | 3 | 5.19 |
| CHEMBL4541637 | — | — | 3 | 5.18 |
| CHEMBL6044376 | — | — | 3 | 5.18 |
| CHEMBL5859408 | — | — | 3 | 5.16 |
| CHEMBL6062840 | — | — | 3 | 5.05 |
| CHEMBL5828860 | — | — | 3 | 5.05 |
| CHEMBL6000630 | — | — | 3 | 5.03 |
| CHEMBL6060287 | — | — | 3 | 5.02 |
| CHEMBL5930150 | — | — | 3 | 4.98 |
| CHEMBL5793130 | — | — | 3 | 4.93 |
| CHEMBL5859591 | — | — | 3 | 4.89 |
| CHEMBL6001759 | — | — | 3 | 4.78 |
| CHEMBL5862685 | — | — | 3 | 4.77 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL4060528 | — | IC50 | = | 322.0 | nM | 6.49 |
| CHEMBL5890455 | — | IC50 | = | 790.0 | nM | 6.10 |
| CHEMBL5923623 | — | IC50 | = | 1530.0 | nM | 5.82 |
| CHEMBL4577393 | — | IC50 | = | 1580.0 | nM | 5.80 |
| CHEMBL4064366 | — | IC50 | = | 1760.0 | nM | 5.75 |
| CHEMBL4085905 | — | IC50 | = | 1770.0 | nM | 5.75 |
| CHEMBL6045935 | — | IC50 | = | 1950.0 | nM | 5.71 |
| CHEMBL5995592 | — | IC50 | = | 2040.0 | nM | 5.69 |
| CHEMBL5912136 | — | IC50 | = | 2530.0 | nM | 5.60 |
| CHEMBL5756048 | — | IC50 | = | 2580.0 | nM | 5.59 |
| CHEMBL5989133 | — | IC50 | = | 3190.0 | nM | 5.50 |
| CHEMBL5748427 | — | IC50 | = | 4220.0 | nM | 5.38 |
| CHEMBL4092807 | — | IC50 | = | 5010.0 | nM | 5.30 |
| CHEMBL5755930 | — | IC50 | = | 5150.0 | nM | 5.29 |
| CHEMBL4450982 | — | IC50 | = | 5370.0 | nM | 5.27 |
| CHEMBL5953308 | — | IC50 | = | 6240.0 | nM | 5.21 |
| CHEMBL5830364 | — | IC50 | = | 6480.0 | nM | 5.19 |
| CHEMBL5785335 | — | IC50 | = | 6390.0 | nM | 5.19 |
| CHEMBL6044376 | — | IC50 | = | 6570.0 | nM | 5.18 |
| CHEMBL4541637 | — | IC50 | = | 6620.0 | nM | 5.18 |
| CHEMBL5859408 | — | IC50 | = | 6950.0 | nM | 5.16 |
| CHEMBL5828860 | — | IC50 | = | 8930.0 | nM | 5.05 |
| CHEMBL6062840 | — | IC50 | = | 8850.0 | nM | 5.05 |
| CHEMBL6000630 | — | IC50 | = | 9260.0 | nM | 5.03 |
| CHEMBL6060287 | — | IC50 | = | 9620.0 | nM | 5.02 |
| CHEMBL5930150 | — | IC50 | = | 10400.0 | nM | 4.98 |
| CHEMBL5793130 | — | IC50 | = | 11800.0 | nM | 4.93 |
| CHEMBL5859591 | — | IC50 | = | 12800.0 | nM | 4.89 |
| CHEMBL6001759 | — | IC50 | = | 16700.0 | nM | 4.78 |
| CHEMBL5862685 | — | IC50 | = | 17000.0 | nM | 4.77 |
| CHEMBL6049497 | — | IC50 | = | 17200.0 | nM | 4.76 |
| CHEMBL6009040 | — | IC50 | = | 17300.0 | nM | 4.76 |
| CHEMBL5838152 | — | IC50 | = | 18100.0 | nM | 4.74 |
| CHEMBL5944377 | — | IC50 | = | 19800.0 | nM | 4.70 |
| CHEMBL5791326 | — | IC50 | = | 20400.0 | nM | 4.69 |
| CHEMBL6034383 | — | IC50 | = | 21100.0 | nM | 4.68 |
| CHEMBL5747451 | — | IC50 | = | 21100.0 | nM | 4.68 |
| CHEMBL5891682 | — | IC50 | = | 22600.0 | nM | 4.65 |
| CHEMBL5742095 | — | IC50 | = | 23600.0 | nM | 4.63 |
| CHEMBL5963863 | — | IC50 | = | 24100.0 | nM | 4.62 |
| CHEMBL5851723 | — | IC50 | = | 25700.0 | nM | 4.59 |
| CHEMBL5982497 | — | IC50 | = | 25500.0 | nM | 4.59 |
| CHEMBL5937523 | — | IC50 | = | 26800.0 | nM | 4.57 |
| CHEMBL6045935 | — | k_off | = | - | s-1 | - |
| CHEMBL6045935 | — | kon | = | - | — | - |
| CHEMBL5756048 | — | k_off | = | - | s-1 | - |
| CHEMBL5887329 | — | kon | = | - | — | - |
| CHEMBL5890455 | — | kon | = | - | — | - |
| CHEMBL5756048 | — | kon | = | - | — | - |
| CHEMBL6034383 | — | kon | = | - | — | - |