Assay Detail
Binding
CHEMBL3887889
Review assay metadata, readout intent, target linkage, and publication context from the same page.
TdF (Temperature Dependence Fluorescence) Assay: The SAR (Structure Activity Relationship) for ERK ligands covered by this invention was interrogated using the TdF (Temperature Dependence Fluorescence) assay or best known as thermal shift assay. (M. W. Pantoliano, et al., "High-density miniaturized thermal shift assays as a general strategy for drug discovery," J. Biomol. Screen 6 (2001) 429-440) The TdF assay was mainly conducted in the 96-well based CHROMO-4 real time fluorescence plate reader (BioRad). The Sypro Orange (Sigma-Aldrich), environmentally sensitive fluorescence dye, was used to monitor the protein folding-unfolding transition. Protein-ligand binding was gauged by the change (or shift) in the unfolding transition temperature (ΔTm) acquired at protein alone with respect to protein in the presence of ligand of interest.Compound of interest was first prepared in DMSO stock (typical concentration: 10 mM). Sample of 20 μt was then added into the 96-well PCR plate, where it consisted of 3 μM ERK protein and 15, 50 or 100 μM compound (depending on compound's solubility) in buffer (25 mM HEPES, 150 mM NaCl, pH-7.5 and 1 mM DTT) incorporated with Sypro Orange dye (5x final concentration). Final percentage of DMSO resided in the sample was 2%. The sample plate was heated from 30° C. to 90° C. with thermal ramping rate of 1° C./min.
22
Total Activities
22
Compounds Tested
1
Activity Types
0
Assay Parameters
Assay Information
| Assay Type | Binding |
| Organism | Mus musculus |
| Confidence | 8 — Homologous single protein target |
| Curated By | Autocuration |
Publication
Fused pyrazole derivatives as novel ERK inhibitors
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| Kd | 22 | 9.46 | 10.52 |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL4108876 | — | — | 1 | 10.52 |
| CHEMBL4112238 | — | — | 1 | 10.52 |
| CHEMBL3982108 | — | — | 1 | 10.22 |
| CHEMBL4110299 | — | — | 1 | 9.92 |
| CHEMBL3963762 | — | — | 1 | 9.89 |
| CHEMBL4106627 | — | — | 1 | 9.85 |
| CHEMBL4109581 | — | — | 1 | 9.82 |
| CHEMBL4111090 | — | — | 1 | 9.82 |
| CHEMBL4108021 | — | — | 1 | 9.60 |
| CHEMBL4107742 | — | — | 1 | 9.52 |
| CHEMBL4115451 | — | — | 1 | 9.51 |
| CHEMBL4114639 | — | — | 1 | 9.43 |
| CHEMBL4107781 | — | — | 1 | 9.40 |
| CHEMBL4110333 | — | — | 1 | 9.33 |
| CHEMBL4111882 | — | — | 1 | 9.29 |
| CHEMBL3966446 | — | — | 1 | 9.15 |
| CHEMBL4114164 | — | — | 1 | 8.95 |
| CHEMBL4111984 | — | — | 1 | 8.87 |
| CHEMBL4108732 | — | — | 1 | 8.82 |
| CHEMBL4114970 | — | — | 1 | 8.81 |
| CHEMBL4108740 | — | — | 1 | 8.68 |
| CHEMBL4110435 | — | — | 1 | 8.22 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL4108876 | — | Kd | = | 0.03 | nM | 10.52 |
| CHEMBL4112238 | — | Kd | = | 0.03 | nM | 10.52 |
| CHEMBL3982108 | — | Kd | = | 0.06 | nM | 10.22 |
| CHEMBL4110299 | — | Kd | = | 0.12 | nM | 9.92 |
| CHEMBL3963762 | — | Kd | = | 0.13 | nM | 9.89 |
| CHEMBL4106627 | — | Kd | = | 0.14 | nM | 9.85 |
| CHEMBL4109581 | — | Kd | = | 0.15 | nM | 9.82 |
| CHEMBL4111090 | — | Kd | = | 0.15 | nM | 9.82 |
| CHEMBL4108021 | — | Kd | = | 0.25 | nM | 9.60 |
| CHEMBL4107742 | — | Kd | = | 0.3 | nM | 9.52 |
| CHEMBL4115451 | — | Kd | = | 0.31 | nM | 9.51 |
| CHEMBL4114639 | — | Kd | = | 0.37 | nM | 9.43 |
| CHEMBL4107781 | — | Kd | = | 0.4 | nM | 9.40 |
| CHEMBL4110333 | — | Kd | = | 0.47 | nM | 9.33 |
| CHEMBL4111882 | — | Kd | = | 0.51 | nM | 9.29 |
| CHEMBL3966446 | — | Kd | = | 0.7 | nM | 9.15 |
| CHEMBL4114164 | — | Kd | = | 1.12 | nM | 8.95 |
| CHEMBL4111984 | — | Kd | = | 1.36 | nM | 8.87 |
| CHEMBL4108732 | — | Kd | = | 1.51 | nM | 8.82 |
| CHEMBL4114970 | — | Kd | = | 1.55 | nM | 8.81 |
| CHEMBL4108740 | — | Kd | = | 2.1 | nM | 8.68 |
| CHEMBL4110435 | — | Kd | = | 6.0 | nM | 8.22 |