Assay Detail
Binding
CHEMBL5733306
Review assay metadata, readout intent, target linkage, and publication context from the same page.
In Vitro Raf Activity: Raf and biotinylated Mek, kinase dead, were combined at 2× final concentrations in assay buffer (50 mM Tris, pH 7.5, 15 mM MgCl2, 0.01% BSA and 1 mM DTT) and dispensed 5 ml per well in assay plates (Greiner white 384 well assay plates #781207) containing 0.25 ml of 40× of a Raf kinase inhibitor test compound diluted in 100% DMSO. The plate was incubated for 60 minutes at room temperature. The Raf kinase activity reaction was started by the addition of 5 mL per well of 2×ATP diluted in assay buffer. After 3 hours (b-Raf(V600E)) or 1 hour (c-Raf). The reactions were stopped and the phosphorylated product was measured using a rabbit anti-p-MEK (Cell Signaling, #9121) antibody and the Alpha Screen IgG (ProteinA) detection Kit (PerkinElmer #6760617R), by the addition of 10 mL to the well of a mixture of the antibody (1:2000 dilution) and detection beads (1:2000 dilution of both beads) in Stop/bead buffer (25 mM EDTA, 50 mM Tris, pH 7.5, 0.01% Tween20). The additions were carried out under dark conditions to protect the detection beads from light. A lid was placed on top of the plate and incubated for 1 hour at room temperature, then the luminescence was read on a PerkinElmer Envision instrument. The concentration of each compound for 50% inhibition (IC50) was calculated by non-linear regression using XL Fit data analysis software.
3819
Total Activities
1057
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 |
Target
RAF proto-oncogene serine/threonine-protein kinase (CHEMBL1906) ↗| Type | SINGLE PROTEIN |
| Organism | Homo sapiens |
Publication
Biaryl amide compounds as kinase inhibitors
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| IC50 | 1273 | 8.99 | 10.40 |
| kon | 1273 | - | - |
| k_off | 1273 | - | - |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL6023111 | — | — | 3 | 10.40 |
| CHEMBL3933272 | — | — | 3 | 10.40 |
| CHEMBL3951474 | — | — | 6 | 10.25 |
| CHEMBL3905257 | — | — | 3 | 10.15 |
| CHEMBL5945139 | — | — | 3 | 10.15 |
| CHEMBL5885861 | — | — | 3 | 10.10 |
| CHEMBL5912922 | — | — | 3 | 10.10 |
| CHEMBL5937628 | — | — | 3 | 10.10 |
| CHEMBL5917557 | — | — | 3 | 10.05 |
| CHEMBL5808970 | — | — | 3 | 10.05 |
| CHEMBL3982146 | — | — | 6 | 10.05 |
| CHEMBL6039326 | — | — | 3 | 10.00 |
| CHEMBL5768029 | — | — | 3 | 10.00 |
| CHEMBL5966098 | — | — | 3 | 10.00 |
| CHEMBL5740520 | — | — | 3 | 10.00 |
| CHEMBL5776436 | — | — | 3 | 10.00 |
| CHEMBL5801973 | — | — | 3 | 10.00 |
| CHEMBL6032960 | — | — | 3 | 10.00 |
| CHEMBL5829780 | — | — | 3 | 10.00 |
| CHEMBL5941581 | — | — | 3 | 10.00 |
| CHEMBL3914464 | — | — | 6 | 10.00 |
| CHEMBL3966766 | — | — | 6 | 10.00 |
| CHEMBL3942721 | — | — | 6 | 10.00 |
| CHEMBL3907015 | — | — | 6 | 10.00 |
| CHEMBL3959581 | — | — | 6 | 10.00 |
| CHEMBL3928030 | — | — | 6 | 10.00 |
| CHEMBL3972141 | — | — | 24 | 10.00 |
| CHEMBL3976881 | — | — | 6 | 10.00 |
| CHEMBL3984282 | — | — | 6 | 10.00 |
| CHEMBL3956654 | — | — | 6 | 10.00 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL3933272 | — | IC50 | = | 0.04 | nM | 10.40 |
| CHEMBL6023111 | — | IC50 | = | 0.04 | nM | 10.40 |
| CHEMBL3951474 | — | IC50 | = | 0.056 | nM | 10.25 |
| CHEMBL3951474 | — | IC50 | = | 0.06 | nM | 10.22 |
| CHEMBL3905257 | — | IC50 | = | 0.07 | nM | 10.15 |
| CHEMBL5945139 | — | IC50 | = | 0.07 | nM | 10.15 |
| CHEMBL5885861 | — | IC50 | = | 0.08 | nM | 10.10 |
| CHEMBL5912922 | — | IC50 | = | 0.08 | nM | 10.10 |
| CHEMBL5937628 | — | IC50 | = | 0.08 | nM | 10.10 |
| CHEMBL3982146 | — | IC50 | = | 0.09 | nM | 10.05 |
| CHEMBL5808970 | — | IC50 | = | 0.09 | nM | 10.05 |
| CHEMBL5917557 | — | IC50 | = | 0.09 | nM | 10.05 |
| CHEMBL5776436 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL3984282 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL5740520 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL5949500 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL5966098 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL6025398 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL5772954 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL3907015 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL5941581 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL3966766 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL3914464 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL5814168 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL3976881 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL3942721 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL5801973 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL6032960 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL6039326 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL5831664 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL3972141 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL3928030 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL3959581 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL5829780 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL3956654 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL5768029 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL5870489 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL5768457 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL5862003 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL3967893 | — | IC50 | = | 0.103 | nM | 9.99 |
| CHEMBL3966766 | — | IC50 | = | 0.103 | nM | 9.99 |
| CHEMBL5993288 | — | IC50 | = | 0.11 | nM | 9.96 |
| CHEMBL3972141 | — | IC50 | = | 0.11 | nM | 9.96 |
| CHEMBL3919205 | — | IC50 | = | 0.117 | nM | 9.93 |
| CHEMBL3962812 | — | IC50 | = | 0.119 | nM | 9.92 |
| CHEMBL5990870 | — | IC50 | = | 0.12 | nM | 9.92 |
| CHEMBL5917765 | — | IC50 | = | 0.12 | nM | 9.92 |
| CHEMBL5758312 | — | IC50 | = | 0.12 | nM | 9.92 |
| CHEMBL3907015 | — | IC50 | = | 0.124 | nM | 9.91 |
| CHEMBL3914464 | — | IC50 | = | 0.122 | nM | 9.91 |