Assay Detail
Binding
CHEMBL3705485
Review assay metadata, readout intent, target linkage, and publication context from the same page.
Mass Spectrometry Assay : The EGLN-1 (or EGLN-3) enzyme activity is determined using mass spectrometry (matrix-assisted laser desorption ionization, time-of-flight MS, MALDI-TOF MS—for assay details, see reference (Greis et al, 2006). The reaction is conducted in a total volume of 50 uL containing TrisCl (5 mM, pH7.5), ascorbate (120 μM), 2-oxoglutarate (3.2 μM), HIF-1α (8.6 μM), and bovine serum albumin (0.01%). The enzyme, quantity predetermined to hydroxylate 20% of substrate in 20 minutes, is added to start the reaction. Compounds are prepared in dimethyl sulfoxide at 10-fold final assay concentration. After 20 minutes at room temperature, the reaction is stopped by transferring 10 μL of reaction mixture to 50 μL of a mass spectrometry matrix solution (α-cyano-4-hydroxycinnamic acid, 5 mg/mL in 50% acetonitrile/0.1% TFA, 5 mM NH4PO4). Two microliters of the mixture is spotted onto a MALDI-TOF MS target plate for analysis with an Applied Biosystems.
25
Total Activities
24
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
Prolyl hydroxylase inhibitors and method of use
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| IC50 | 25 | 5.79 | 6.72 |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL5897931 | — | — | 1 | 6.72 |
| CHEMBL3646220 | — | — | 1 | 6.62 |
| CHEMBL3646233 | — | — | 1 | 6.52 |
| CHEMBL3646224 | — | — | 1 | 6.52 |
| CHEMBL3646234 | — | — | 1 | 6.40 |
| CHEMBL3646221 | VADADUSTAT | 4.0 | 2 | 6.39 |
| CHEMBL3646223 | — | — | 1 | 6.36 |
| CHEMBL3646215 | — | — | 1 | 6.19 |
| CHEMBL3646222 | — | — | 1 | 5.96 |
| CHEMBL3646227 | — | — | 1 | 5.92 |
| CHEMBL3646225 | — | — | 1 | 5.80 |
| CHEMBL3646232 | — | — | 1 | 5.60 |
| CHEMBL3646226 | — | — | 1 | 5.60 |
| CHEMBL3646229 | — | — | 1 | 5.58 |
| CHEMBL3646212 | — | — | 1 | 5.55 |
| CHEMBL3646213 | — | — | 1 | 5.43 |
| CHEMBL3646217 | — | — | 1 | 5.39 |
| CHEMBL3646228 | — | — | 1 | 5.29 |
| CHEMBL3646214 | — | — | 1 | 5.24 |
| CHEMBL3646218 | — | — | 1 | 5.17 |
| CHEMBL3646235 | — | — | 1 | 5.04 |
| CHEMBL3646231 | — | — | 1 | 4.95 |
| CHEMBL3646216 | — | — | 1 | 4.70 |
| CHEMBL3646219 | — | — | 1 | - |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL5897931 | — | IC50 | = | 190.0 | nM | 6.72 |
| CHEMBL3646220 | — | IC50 | = | 240.0 | nM | 6.62 |
| CHEMBL3646233 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL3646224 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL3646234 | — | IC50 | = | 400.0 | nM | 6.40 |
| CHEMBL3646221 | VADADUSTAT | IC50 | = | 410.0 | nM | 6.39 |
| CHEMBL3646223 | — | IC50 | = | 440.0 | nM | 6.36 |
| CHEMBL3646215 | — | IC50 | = | 650.0 | nM | 6.19 |
| CHEMBL3646222 | — | IC50 | = | 1100.0 | nM | 5.96 |
| CHEMBL3646221 | VADADUSTAT | IC50 | = | 1100.0 | nM | 5.96 |
| CHEMBL3646227 | — | IC50 | = | 1200.0 | nM | 5.92 |
| CHEMBL3646225 | — | IC50 | = | 1600.0 | nM | 5.80 |
| CHEMBL3646226 | — | IC50 | = | 2500.0 | nM | 5.60 |
| CHEMBL3646232 | — | IC50 | = | 2500.0 | nM | 5.60 |
| CHEMBL3646229 | — | IC50 | = | 2600.0 | nM | 5.58 |
| CHEMBL3646212 | — | IC50 | = | 2800.0 | nM | 5.55 |
| CHEMBL3646213 | — | IC50 | = | 3700.0 | nM | 5.43 |
| CHEMBL3646217 | — | IC50 | = | 4100.0 | nM | 5.39 |
| CHEMBL3646228 | — | IC50 | = | 5100.0 | nM | 5.29 |
| CHEMBL3646214 | — | IC50 | = | 5800.0 | nM | 5.24 |
| CHEMBL3646218 | — | IC50 | = | 6800.0 | nM | 5.17 |
| CHEMBL3646235 | — | IC50 | = | 9200.0 | nM | 5.04 |
| CHEMBL3646231 | — | IC50 | = | 11200.0 | nM | 4.95 |
| CHEMBL3646216 | — | IC50 | = | 20000.0 | nM | 4.70 |
| CHEMBL3646219 | — | IC50 | = | 99000.0 | nM | - |