Assay Detail
Binding
CHEMBL5737798
Review assay metadata, readout intent, target linkage, and publication context from the same page.
Measurement of RET Inhibitory Activity (In Vitro): Regarding the conditions for measurement of in vitro inhibitory activity of compounds against RET kinase activity, the website of AnaSpec states that Srctide (GEEPLYWSFPAKKK) corresponds to the substrate peptide for reaction to measure RET kinase activity. Thus, the amino acid sequence was partly modified and biotinylated to prepare biotinylated peptides (biotin-EEPLYWSFPAKKK). The purified recombinant human RET protein used in the test was purchased from Carna Biosciences, Inc.To measure the inhibitory activity, first, the compounds of the present invention were individually diluted with dimethyl sulfoxide (DMSO) stepwise. Subsequently, RET protein, the substrate peptide (final concentration: 250 nM), magnesium chloride (final concentration: 10 mM), ATP (the final concentration: 10 μM), and a solution of the compound of the present invention in DMSO (final concentration of DMSO: 2.5%) were added to a buffer for kinase reaction (13.5 mM Tris, pH of 7.5, 2 mM dithiothreitol, 0.009% Tween-20). Each of the mixtures was incubated at 25° C. for 100 minutes to perform kinase reaction. EDTA was then added thereto to give a final concentration of 24 mM so that the reaction was terminated. A detection solution containing Eu-labeled antiphosphotyrosine antibody PT66 (PerkinElmer) and SureLight APC-SA (PerkinElmer) was added thereto, and each mixture was allowed to stand at room temperature for 2 hours or more. Finally, the intensity of fluorescence under the excitation light with a wavelength of 337 nm was measured with a PHERAstar FS (BMG Labtech) at the two wavelengths of 620 nm and 665 nm. The phosphorylation level was calculated from the ratio of the fluorescence intensity at the two wavelengths, and the compound concentration at which phosphorylation was inhibited by 50% was defined as the IC50 value (nM).
234
Total Activities
78
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
Proto-oncogene tyrosine-protein kinase receptor Ret (CHEMBL2041) ↗| Type | SINGLE PROTEIN |
| Organism | Homo sapiens |
Publication
Fused pyrimidine compound or salt thereof
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| IC50 | 78 | 8.97 | 10.40 |
| kon | 78 | - | - |
| k_off | 78 | - | - |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL5944764 | — | — | 3 | 10.40 |
| CHEMBL5916213 | — | — | 3 | 10.15 |
| CHEMBL5838635 | — | — | 3 | 10.05 |
| CHEMBL6035624 | — | — | 3 | 10.00 |
| CHEMBL5902423 | — | — | 3 | 10.00 |
| CHEMBL5807109 | — | — | 3 | 9.85 |
| CHEMBL5895474 | — | — | 3 | 9.82 |
| CHEMBL5747405 | — | — | 3 | 9.82 |
| CHEMBL5909127 | — | — | 3 | 9.82 |
| CHEMBL5753786 | — | — | 3 | 9.82 |
| CHEMBL5876102 | — | — | 3 | 9.82 |
| CHEMBL5787119 | — | — | 3 | 9.82 |
| CHEMBL5992123 | — | — | 3 | 9.80 |
| CHEMBL5982797 | — | — | 3 | 9.77 |
| CHEMBL5747090 | — | — | 3 | 9.77 |
| CHEMBL5887050 | — | — | 3 | 9.70 |
| CHEMBL5872379 | — | — | 3 | 9.68 |
| CHEMBL5786868 | — | — | 3 | 9.64 |
| CHEMBL6060970 | — | — | 3 | 9.64 |
| CHEMBL5879224 | — | — | 3 | 9.57 |
| CHEMBL5982088 | — | — | 3 | 9.57 |
| CHEMBL6005276 | — | — | 3 | 9.55 |
| CHEMBL5969042 | — | — | 3 | 9.54 |
| CHEMBL5916732 | — | — | 3 | 9.49 |
| CHEMBL5868962 | — | — | 3 | 9.47 |
| CHEMBL5958139 | — | — | 3 | 9.44 |
| CHEMBL5930401 | — | — | 3 | 9.40 |
| CHEMBL5901100 | — | — | 3 | 9.40 |
| CHEMBL5923647 | — | — | 3 | 9.37 |
| CHEMBL5918408 | — | — | 3 | 9.34 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL5944764 | — | IC50 | = | 0.04 | nM | 10.40 |
| CHEMBL5916213 | — | IC50 | = | 0.07 | nM | 10.15 |
| CHEMBL5838635 | — | IC50 | = | 0.09 | nM | 10.05 |
| CHEMBL6035624 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL5902423 | — | IC50 | = | 0.1 | nM | 10.00 |
| CHEMBL5807109 | — | IC50 | = | 0.14 | nM | 9.85 |
| CHEMBL5895474 | — | IC50 | = | 0.15 | nM | 9.82 |
| CHEMBL5876102 | — | IC50 | = | 0.15 | nM | 9.82 |
| CHEMBL5747405 | — | IC50 | = | 0.15 | nM | 9.82 |
| CHEMBL5787119 | — | IC50 | = | 0.15 | nM | 9.82 |
| CHEMBL5909127 | — | IC50 | = | 0.15 | nM | 9.82 |
| CHEMBL5753786 | — | IC50 | = | 0.15 | nM | 9.82 |
| CHEMBL5992123 | — | IC50 | = | 0.16 | nM | 9.80 |
| CHEMBL5747090 | — | IC50 | = | 0.17 | nM | 9.77 |
| CHEMBL5982797 | — | IC50 | = | 0.17 | nM | 9.77 |
| CHEMBL5887050 | — | IC50 | = | 0.2 | nM | 9.70 |
| CHEMBL5872379 | — | IC50 | = | 0.21 | nM | 9.68 |
| CHEMBL5786868 | — | IC50 | = | 0.23 | nM | 9.64 |
| CHEMBL6060970 | — | IC50 | = | 0.23 | nM | 9.64 |
| CHEMBL5879224 | — | IC50 | = | 0.27 | nM | 9.57 |
| CHEMBL5982088 | — | IC50 | = | 0.266 | nM | 9.57 |
| CHEMBL6005276 | — | IC50 | = | 0.279 | nM | 9.55 |
| CHEMBL5969042 | — | IC50 | = | 0.29 | nM | 9.54 |
| CHEMBL5916732 | — | IC50 | = | 0.32 | nM | 9.49 |
| CHEMBL5868962 | — | IC50 | = | 0.337 | nM | 9.47 |
| CHEMBL5958139 | — | IC50 | = | 0.36 | nM | 9.44 |
| CHEMBL5930401 | — | IC50 | = | 0.4 | nM | 9.40 |
| CHEMBL5901100 | — | IC50 | = | 0.4 | nM | 9.40 |
| CHEMBL5923647 | — | IC50 | = | 0.43 | nM | 9.37 |
| CHEMBL5918408 | — | IC50 | = | 0.46 | nM | 9.34 |
| CHEMBL5939730 | — | IC50 | = | 0.48 | nM | 9.32 |
| CHEMBL5890640 | — | IC50 | = | 0.51 | nM | 9.29 |
| CHEMBL5746968 | — | IC50 | = | 0.59 | nM | 9.23 |
| CHEMBL5987493 | — | IC50 | = | 0.61 | nM | 9.21 |
| CHEMBL5741664 | — | IC50 | = | 0.61 | nM | 9.21 |
| CHEMBL5845614 | — | IC50 | = | 0.76 | nM | 9.12 |
| CHEMBL5760086 | — | IC50 | = | 0.788 | nM | 9.10 |
| CHEMBL6026130 | — | IC50 | = | 0.79 | nM | 9.10 |
| CHEMBL5811565 | — | IC50 | = | 0.83 | nM | 9.08 |
| CHEMBL5817133 | — | IC50 | = | 1.0 | nM | 9.00 |
| CHEMBL6049914 | — | IC50 | = | 1.0 | nM | 9.00 |
| CHEMBL5800562 | — | IC50 | = | 1.07 | nM | 8.97 |
| CHEMBL5977421 | — | IC50 | = | 1.35 | nM | 8.87 |
| CHEMBL5804877 | — | IC50 | = | 1.47 | nM | 8.83 |
| CHEMBL5964862 | — | IC50 | = | 1.48 | nM | 8.83 |
| CHEMBL5821817 | — | IC50 | = | 1.53 | nM | 8.81 |
| CHEMBL5829996 | — | IC50 | = | 1.73 | nM | 8.76 |
| CHEMBL5749302 | — | IC50 | = | 1.86 | nM | 8.73 |
| CHEMBL5877425 | — | IC50 | = | 1.86 | nM | 8.73 |
| CHEMBL5837715 | — | IC50 | = | 1.94 | nM | 8.71 |