Assay Detail
Binding
CHEMBL5737408
Review assay metadata, readout intent, target linkage, and publication context from the same page.
Enzymatic Assay2: The enzymatic assay couples DHODH activity with bleaching of the dye 2,6-Dichlorophenolindophenol (DCIP) (Knecht and Loffler, 1998; Miller et al., 1968). The assay was conducted in aqueous buffer containing 50 mM Tris, 0.1% Triton X-100, 150 mM potassium chloride, 0.4 μg/mL DHODH, 1 mM dihydroorotate, 0.1 mM decylubiquinone, 0.06 mM DCIP, and 0.17% DMSO at pH 8.0 at room temperature. Compounds were added via pin transfer or via D300 digital dispenser, and the reaction was initiated by addition of substrates. Enzyme activity was monitored kinetically by the reduction in DCIP absorbance at 600 nm. Purified recombinant human DHODH (full-length, C-terminal MYC/DDK-tag) enzyme was purchased from Origene (cat. no. TP039034). Other chemicals, including leflunomide and teriflunomide, were purchased from Sigma-Aldrich. Absorbance measurements were obtained using a Molecular Devices Spectramax M5 plate-reading spectrophotometer.
210
Total Activities
70
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
Dihydroorotate dehydrogenase (quinone), mitochondrial (CHEMBL1966) ↗| Type | SINGLE PROTEIN |
| Organism | Homo sapiens |
Publication
2,4,5-trisubstituted 1,2,4-triazolones useful as inhibitors of DHODH
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| IC50 | 70 | 6.48 | 9.00 |
| kon | 70 | - | - |
| k_off | 70 | - | - |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL5837805 | — | — | 3 | 9.00 |
| CHEMBL5910343 | — | — | 3 | 8.74 |
| CHEMBL5961667 | — | — | 3 | 8.55 |
| CHEMBL5780640 | — | — | 3 | 8.52 |
| CHEMBL6025330 | — | — | 3 | 8.27 |
| CHEMBL6057870 | — | — | 3 | 8.26 |
| CHEMBL5882889 | — | — | 3 | 8.10 |
| CHEMBL6062298 | — | — | 3 | 8.05 |
| CHEMBL5987725 | — | — | 3 | 7.96 |
| CHEMBL5888619 | — | — | 3 | 7.85 |
| CHEMBL5854933 | — | — | 3 | 7.82 |
| CHEMBL5891802 | — | — | 3 | 7.77 |
| CHEMBL5953067 | — | — | 3 | 7.77 |
| CHEMBL5898676 | — | — | 3 | 7.72 |
| CHEMBL6032087 | — | — | 3 | 7.68 |
| CHEMBL5986568 | — | — | 3 | 7.67 |
| CHEMBL5742229 | — | — | 3 | 7.65 |
| CHEMBL6030009 | — | — | 3 | 7.42 |
| CHEMBL5871444 | — | — | 3 | 7.38 |
| CHEMBL5858428 | — | — | 3 | 7.28 |
| CHEMBL6017108 | — | — | 3 | 7.26 |
| CHEMBL5753736 | — | — | 3 | 7.25 |
| CHEMBL6061337 | — | — | 3 | 7.19 |
| CHEMBL5758058 | — | — | 3 | 7.14 |
| CHEMBL5900744 | — | — | 3 | 7.09 |
| CHEMBL5916952 | — | — | 3 | 7.07 |
| CHEMBL6021562 | — | — | 3 | 7.03 |
| CHEMBL6036019 | — | — | 3 | 6.98 |
| CHEMBL6017382 | — | — | 3 | 6.83 |
| CHEMBL5922631 | — | — | 3 | 6.82 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL5837805 | — | IC50 | = | 1.0 | nM | 9.00 |
| CHEMBL5910343 | — | IC50 | = | 1.8 | nM | 8.74 |
| CHEMBL5961667 | — | IC50 | = | 2.8 | nM | 8.55 |
| CHEMBL5780640 | — | IC50 | = | 3.0 | nM | 8.52 |
| CHEMBL6025330 | — | IC50 | = | 5.4 | nM | 8.27 |
| CHEMBL6057870 | — | IC50 | = | 5.5 | nM | 8.26 |
| CHEMBL5882889 | — | IC50 | = | 8.0 | nM | 8.10 |
| CHEMBL6062298 | — | IC50 | = | 9.0 | nM | 8.05 |
| CHEMBL5987725 | — | IC50 | = | 11.0 | nM | 7.96 |
| CHEMBL5888619 | — | IC50 | = | 14.0 | nM | 7.85 |
| CHEMBL5854933 | — | IC50 | = | 15.3 | nM | 7.82 |
| CHEMBL5953067 | — | IC50 | = | 17.0 | nM | 7.77 |
| CHEMBL5891802 | — | IC50 | = | 17.0 | nM | 7.77 |
| CHEMBL5898676 | — | IC50 | = | 19.0 | nM | 7.72 |
| CHEMBL6032087 | — | IC50 | = | 21.0 | nM | 7.68 |
| CHEMBL5986568 | — | IC50 | = | 21.2 | nM | 7.67 |
| CHEMBL5742229 | — | IC50 | = | 22.4 | nM | 7.65 |
| CHEMBL6030009 | — | IC50 | = | 37.7 | nM | 7.42 |
| CHEMBL5871444 | — | IC50 | = | 41.5 | nM | 7.38 |
| CHEMBL5858428 | — | IC50 | = | 52.0 | nM | 7.28 |
| CHEMBL6017108 | — | IC50 | = | 54.6 | nM | 7.26 |
| CHEMBL5753736 | — | IC50 | = | 56.8 | nM | 7.25 |
| CHEMBL6061337 | — | IC50 | = | 64.3 | nM | 7.19 |
| CHEMBL5758058 | — | IC50 | = | 71.6 | nM | 7.14 |
| CHEMBL5900744 | — | IC50 | = | 80.5 | nM | 7.09 |
| CHEMBL5916952 | — | IC50 | = | 85.7 | nM | 7.07 |
| CHEMBL6021562 | — | IC50 | = | 92.9 | nM | 7.03 |
| CHEMBL6036019 | — | IC50 | = | 104.0 | nM | 6.98 |
| CHEMBL6017382 | — | IC50 | = | 149.0 | nM | 6.83 |
| CHEMBL5922631 | — | IC50 | = | 152.0 | nM | 6.82 |
| CHEMBL6053874 | — | IC50 | = | 190.0 | nM | 6.72 |
| CHEMBL6034462 | — | IC50 | = | 213.0 | nM | 6.67 |
| CHEMBL5871141 | — | IC50 | = | 219.0 | nM | 6.66 |
| CHEMBL5994483 | — | IC50 | = | 246.0 | nM | 6.61 |
| CHEMBL5801349 | — | IC50 | = | 252.0 | nM | 6.60 |
| CHEMBL5800658 | — | IC50 | = | 289.0 | nM | 6.54 |
| CHEMBL5754716 | — | IC50 | = | 295.0 | nM | 6.53 |
| CHEMBL5991000 | — | IC50 | = | 409.0 | nM | 6.39 |
| CHEMBL5897487 | — | IC50 | = | 418.0 | nM | 6.38 |
| CHEMBL6065377 | — | IC50 | = | 665.0 | nM | 6.18 |
| CHEMBL5767805 | — | IC50 | = | 738.0 | nM | 6.13 |
| CHEMBL5878423 | — | IC50 | = | 755.0 | nM | 6.12 |
| CHEMBL6032510 | — | IC50 | = | 849.0 | nM | 6.07 |
| CHEMBL5753829 | — | IC50 | = | 867.0 | nM | 6.06 |
| CHEMBL5875474 | — | IC50 | = | 1000.0 | nM | 6.00 |
| CHEMBL5956604 | — | IC50 | = | 1100.0 | nM | 5.96 |
| CHEMBL5864771 | — | IC50 | = | 1100.0 | nM | 5.96 |
| CHEMBL5798342 | — | IC50 | = | 1100.0 | nM | 5.96 |
| CHEMBL6035903 | — | IC50 | = | 1700.0 | nM | 5.77 |
| CHEMBL5991714 | — | IC50 | = | 2000.0 | nM | 5.70 |