Assay Detail
Binding
CHEMBL5735193
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)). 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)). 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.).
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 | 6.98 | 8.40 |
| kon | 69 | - | - |
| k_off | 69 | - | - |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL5923623 | — | — | 3 | 8.40 |
| CHEMBL5912136 | — | — | 3 | 8.15 |
| CHEMBL4064366 | — | — | 3 | 8.15 |
| CHEMBL4577393 | — | — | 3 | 8.10 |
| CHEMBL5785335 | — | — | 3 | 8.05 |
| CHEMBL4450982 | — | — | 3 | 8.00 |
| CHEMBL5859408 | — | — | 3 | 7.89 |
| CHEMBL5830364 | — | — | 3 | 7.89 |
| CHEMBL5944377 | — | — | 3 | 7.82 |
| CHEMBL6062840 | — | — | 3 | 7.77 |
| CHEMBL5953308 | — | — | 3 | 7.72 |
| CHEMBL4541637 | — | — | 3 | 7.72 |
| CHEMBL5862685 | — | — | 3 | 7.70 |
| CHEMBL5930150 | — | — | 3 | 7.62 |
| CHEMBL5828860 | — | — | 3 | 7.62 |
| CHEMBL5748427 | — | — | 3 | 7.57 |
| CHEMBL5755930 | — | — | 3 | 7.55 |
| CHEMBL5851723 | — | — | 3 | 7.54 |
| CHEMBL6060287 | — | — | 3 | 7.52 |
| CHEMBL6028717 | — | — | 3 | 7.40 |
| CHEMBL6044376 | — | — | 3 | 7.37 |
| CHEMBL6049497 | — | — | 3 | 7.29 |
| CHEMBL5982497 | — | — | 3 | 7.26 |
| CHEMBL6009040 | — | — | 3 | 7.24 |
| CHEMBL5891682 | — | — | 3 | 7.17 |
| CHEMBL6000630 | — | — | 3 | 7.16 |
| CHEMBL6001759 | — | — | 3 | 7.16 |
| CHEMBL5869385 | — | — | 3 | 7.16 |
| CHEMBL4577523 | GOLIDOCITINIB | 2.0 | 3 | 7.14 |
| CHEMBL5747451 | — | — | 3 | 7.10 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL5923623 | — | IC50 | = | 4.0 | nM | 8.40 |
| CHEMBL5912136 | — | IC50 | = | 7.0 | nM | 8.15 |
| CHEMBL4064366 | — | IC50 | = | 7.0 | nM | 8.15 |
| CHEMBL4577393 | — | IC50 | = | 8.0 | nM | 8.10 |
| CHEMBL5785335 | — | IC50 | = | 9.0 | nM | 8.05 |
| CHEMBL4450982 | — | IC50 | = | 10.0 | nM | 8.00 |
| CHEMBL5830364 | — | IC50 | = | 13.0 | nM | 7.89 |
| CHEMBL5859408 | — | IC50 | = | 13.0 | nM | 7.89 |
| CHEMBL5944377 | — | IC50 | = | 15.0 | nM | 7.82 |
| CHEMBL6062840 | — | IC50 | = | 17.0 | nM | 7.77 |
| CHEMBL5953308 | — | IC50 | = | 19.0 | nM | 7.72 |
| CHEMBL4541637 | — | IC50 | = | 19.0 | nM | 7.72 |
| CHEMBL5862685 | — | IC50 | = | 20.0 | nM | 7.70 |
| CHEMBL5828860 | — | IC50 | = | 24.0 | nM | 7.62 |
| CHEMBL5930150 | — | IC50 | = | 24.0 | nM | 7.62 |
| CHEMBL5748427 | — | IC50 | = | 27.0 | nM | 7.57 |
| CHEMBL5755930 | — | IC50 | = | 28.0 | nM | 7.55 |
| CHEMBL5851723 | — | IC50 | = | 29.0 | nM | 7.54 |
| CHEMBL6060287 | — | IC50 | = | 30.0 | nM | 7.52 |
| CHEMBL6028717 | — | IC50 | = | 40.0 | nM | 7.40 |
| CHEMBL6044376 | — | IC50 | = | 43.0 | nM | 7.37 |
| CHEMBL6049497 | — | IC50 | = | 51.0 | nM | 7.29 |
| CHEMBL5982497 | — | IC50 | = | 55.0 | nM | 7.26 |
| CHEMBL6009040 | — | IC50 | = | 57.0 | nM | 7.24 |
| CHEMBL5891682 | — | IC50 | = | 68.0 | nM | 7.17 |
| CHEMBL6001759 | — | IC50 | = | 69.0 | nM | 7.16 |
| CHEMBL5869385 | — | IC50 | = | 69.0 | nM | 7.16 |
| CHEMBL6000630 | — | IC50 | = | 69.0 | nM | 7.16 |
| CHEMBL4577523 | GOLIDOCITINIB | IC50 | = | 73.0 | nM | 7.14 |
| CHEMBL5747451 | — | IC50 | = | 79.0 | nM | 7.10 |
| CHEMBL5859591 | — | IC50 | = | 85.0 | nM | 7.07 |
| CHEMBL5791326 | — | IC50 | = | 97.0 | nM | 7.01 |
| CHEMBL5793130 | — | IC50 | = | 106.0 | nM | 6.97 |
| CHEMBL5838152 | — | IC50 | = | 107.0 | nM | 6.97 |
| CHEMBL6034383 | — | IC50 | = | 106.0 | nM | 6.97 |
| CHEMBL5742095 | — | IC50 | = | 110.0 | nM | 6.96 |
| CHEMBL5852498 | — | IC50 | = | 113.0 | nM | 6.95 |
| CHEMBL5747892 | — | IC50 | = | 166.0 | nM | 6.78 |
| CHEMBL5769895 | — | IC50 | = | 184.0 | nM | 6.74 |
| CHEMBL5937523 | — | IC50 | = | 191.0 | nM | 6.72 |
| CHEMBL5887329 | — | IC50 | = | 192.0 | nM | 6.72 |
| CHEMBL5963863 | — | IC50 | = | 265.0 | nM | 6.58 |
| CHEMBL5879809 | — | IC50 | = | 272.0 | nM | 6.57 |
| CHEMBL5765520 | — | IC50 | = | 296.0 | nM | 6.53 |
| CHEMBL6043947 | — | IC50 | = | 296.0 | nM | 6.53 |
| CHEMBL5748446 | — | IC50 | = | 304.0 | nM | 6.52 |
| CHEMBL5873822 | — | IC50 | = | 359.0 | nM | 6.45 |
| CHEMBL5787931 | — | IC50 | = | 426.0 | nM | 6.37 |
| CHEMBL5979824 | — | IC50 | = | 454.0 | nM | 6.34 |
| CHEMBL6055526 | — | IC50 | = | 490.0 | nM | 6.31 |