Assay Detail
Binding
CHEMBL3705477
Review assay metadata, readout intent, target linkage, and publication context from the same page.
Fluorescence Resonance Energy Transfer (FRET) Assay: The enzyme inhibitory activity of test compounds was determined by fluorescence resonance energy transfer (FRET) using synthetic peptides labelled with both donor and acceptor fluorophores (Z-LYTE, Invitrogen). Recombinant, phosphorylated p38 MAPK (MAPK12:Millipore) was diluted in HEPES buffer, mixed with compound at the desired final concentrations and incubated for 2 hr at RT. The FRET peptide (2 uM) and ATP (100 uM) were next added to the enzyme/compound mixture and incubated for 1 hr. Development reagent (protease) was added for 1 hr prior to detection in a fluorescence microplate reader. The site-specific protease only cleaves non-phosphorylated peptide and eliminates the FRET signal. Phosphorylation levels of each reaction were calculated using the ratio of coumarin emission (donor) over fluorescein emission (acceptor) with high ratios indicating high phosphorylation levels and low ratios indicating low phosphorylation levels.
37
Total Activities
36
Compounds Tested
1
Activity Types
0
Assay Parameters
Assay Information
| Assay Type | Binding |
| Organism | Homo sapiens |
| Confidence | 9 — Direct single protein target |
| Curated By | Autocuration |
Publication
Respiratory formulations and compounds for use therein
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| IC50 | 37 | 8.25 | 9.00 |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL1090979 | — | — | 1 | 9.00 |
| CHEMBL3695591 | — | — | 1 | 8.70 |
| CHEMBL3695576 | — | — | 1 | 8.70 |
| CHEMBL3695584 | — | — | 1 | 8.70 |
| CHEMBL1214131 | — | — | 1 | 8.70 |
| CHEMBL3695578 | — | — | 1 | 8.70 |
| CHEMBL3695593 | — | — | 1 | 8.70 |
| CHEMBL3695575 | — | — | 1 | 8.52 |
| CHEMBL1214134 | — | — | 1 | 8.52 |
| CHEMBL1214132 | — | — | 1 | 8.52 |
| CHEMBL3695588 | — | — | 1 | 8.40 |
| CHEMBL3695580 | — | — | 1 | 8.40 |
| CHEMBL3695573 | — | — | 2 | 8.40 |
| CHEMBL3695590 | — | — | 1 | 8.30 |
| CHEMBL3695581 | — | — | 1 | 8.30 |
| CHEMBL3695579 | — | — | 1 | 8.30 |
| CHEMBL3695571 | — | — | 1 | 8.30 |
| CHEMBL3695577 | — | — | 1 | 8.30 |
| CHEMBL1093756 | — | — | 1 | 8.30 |
| CHEMBL3695568 | JNJ-49095397 | 2.0 | 1 | 8.30 |
| CHEMBL3695583 | — | — | 1 | 8.22 |
| CHEMBL3695592 | — | — | 1 | 8.22 |
| CHEMBL1214135 | — | — | 1 | 8.15 |
| CHEMBL3695589 | — | — | 1 | 8.15 |
| CHEMBL3695572 | — | — | 1 | 8.15 |
| CHEMBL3695574 | — | — | 1 | 8.15 |
| CHEMBL3695586 | — | — | 1 | 8.15 |
| CHEMBL3695585 | — | — | 1 | 7.96 |
| CHEMBL103667 | DORAMAPIMOD | 2.0 | 1 | 7.92 |
| CHEMBL3695569 | — | — | 1 | 7.92 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL1090979 | — | IC50 | = | 1.0 | nM | 9.00 |
| CHEMBL3695591 | — | IC50 | = | 2.0 | nM | 8.70 |
| CHEMBL3695576 | — | IC50 | = | 2.0 | nM | 8.70 |
| CHEMBL1214131 | — | IC50 | = | 2.0 | nM | 8.70 |
| CHEMBL3695584 | — | IC50 | = | 2.0 | nM | 8.70 |
| CHEMBL3695578 | — | IC50 | = | 2.0 | nM | 8.70 |
| CHEMBL3695593 | — | IC50 | = | 2.0 | nM | 8.70 |
| CHEMBL1214132 | — | IC50 | = | 3.0 | nM | 8.52 |
| CHEMBL3695575 | — | IC50 | = | 3.0 | nM | 8.52 |
| CHEMBL1214134 | — | IC50 | = | 3.0 | nM | 8.52 |
| CHEMBL3695588 | — | IC50 | = | 4.0 | nM | 8.40 |
| CHEMBL3695573 | — | IC50 | = | 4.0 | nM | 8.40 |
| CHEMBL3695580 | — | IC50 | = | 4.0 | nM | 8.40 |
| CHEMBL3695581 | — | IC50 | = | 5.0 | nM | 8.30 |
| CHEMBL3695568 | JNJ-49095397 | IC50 | = | 5.0 | nM | 8.30 |
| CHEMBL1093756 | — | IC50 | = | 5.0 | nM | 8.30 |
| CHEMBL3695590 | — | IC50 | = | 5.0 | nM | 8.30 |
| CHEMBL3695579 | — | IC50 | = | 5.0 | nM | 8.30 |
| CHEMBL3695577 | — | IC50 | = | 5.0 | nM | 8.30 |
| CHEMBL3695571 | — | IC50 | = | 5.0 | nM | 8.30 |
| CHEMBL3695583 | — | IC50 | = | 6.0 | nM | 8.22 |
| CHEMBL3695592 | — | IC50 | = | 6.0 | nM | 8.22 |
| CHEMBL3695586 | — | IC50 | = | 7.0 | nM | 8.15 |
| CHEMBL3695572 | — | IC50 | = | 7.0 | nM | 8.15 |
| CHEMBL3695589 | — | IC50 | = | 7.0 | nM | 8.15 |
| CHEMBL1214135 | — | IC50 | = | 7.0 | nM | 8.15 |
| CHEMBL3695574 | — | IC50 | = | 7.0 | nM | 8.15 |
| CHEMBL3695585 | — | IC50 | = | 11.0 | nM | 7.96 |
| CHEMBL103667 | DORAMAPIMOD | IC50 | = | 12.0 | nM | 7.92 |
| CHEMBL3695569 | — | IC50 | = | 12.0 | nM | 7.92 |
| CHEMBL3695594 | — | IC50 | = | 12.0 | nM | 7.92 |
| CHEMBL3695595 | — | IC50 | = | 17.0 | nM | 7.77 |
| CHEMBL1090975 | — | IC50 | = | 17.0 | nM | 7.77 |
| CHEMBL3695573 | — | IC50 | = | 17.0 | nM | 7.77 |
| CHEMBL3695570 | — | IC50 | = | 18.0 | nM | 7.75 |
| CHEMBL3695582 | — | IC50 | = | 21.0 | nM | 7.68 |
| CHEMBL3695587 | — | IC50 | = | 30.0 | nM | 7.52 |