Assay Detail
Binding
CHEMBL5731270
Review assay metadata, readout intent, target linkage, and publication context from the same page.
KDM5 Enzyme Assay: Full length KDM5A enzyme was expressed and purified inhouse. Biotin-H3K4me3 peptide was purchased from New England Biolabs. HTRF reagents (containing Eu-labeled H3K4me1-2 antibody, and streptavidin-XL665) were purchased from Cis-Bio International. Plates were read on an Envision multi-label plate reader (Perkin Elmer).The HTRF assay mixture contained 2 nM full length KDM5A enzyme, 100 nM H3K4Me3 peptide substrate, 1 uM 2-OG, 100 uM Fe2+, 500 uM ascorbate. 50 mM HEPES pH 7.0 buffer, 0.01% Triton-X 100, 2 mM DTT, 0.25% DMSO at a final volume of 10 uL.The enzyme reaction was carried out at room temperature in black Proxiplate 384-Plus plate (Corning. Costar) within 30 minutes, in the presence of varying concentration of a test compound. At the end of enzyme reaction, 5 uL of 1 mM EDTA were added to quench the reaction and then the detection reagents (5 uL) were added to give final concentrations of 0.5 nM Eu-labeled H3K4me1-2 antibody, and 50 nM streptavidin-XL665. The plates were incubated at room temperature for 60 minutes and then read in the Envision plate reader. The readouts were transformed into % inhibition, and IC50 value of a test compounds was generated by using four parameters curve fitting (Model 205 in XLFIT5, iDBS).
738
Total Activities
243
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 |
Publication
Pyrrolidine amide compounds as histone demethylase inhibitors
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| IC50 | 246 | 9.25 | 10.60 |
| kon | 246 | - | - |
| k_off | 246 | - | - |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL5945932 | — | — | 3 | 10.60 |
| CHEMBL6053239 | — | — | 3 | 10.43 |
| CHEMBL5908882 | — | — | 3 | 10.42 |
| CHEMBL5784930 | — | — | 3 | 10.41 |
| CHEMBL5877919 | — | — | 3 | 10.34 |
| CHEMBL5767733 | — | — | 3 | 10.33 |
| CHEMBL4088737 | — | — | 3 | 10.31 |
| CHEMBL5798182 | — | — | 3 | 10.30 |
| CHEMBL5982539 | — | — | 3 | 10.29 |
| CHEMBL5810582 | — | — | 3 | 10.29 |
| CHEMBL5769925 | — | — | 3 | 10.28 |
| CHEMBL5857962 | — | — | 3 | 10.27 |
| CHEMBL5893489 | — | — | 3 | 10.24 |
| CHEMBL6049070 | — | — | 3 | 10.24 |
| CHEMBL6040479 | — | — | 3 | 10.23 |
| CHEMBL5826403 | — | — | 3 | 10.19 |
| CHEMBL5963139 | — | — | 3 | 10.18 |
| CHEMBL5931344 | — | — | 3 | 10.18 |
| CHEMBL6063394 | — | — | 3 | 10.15 |
| CHEMBL5820582 | — | — | 3 | 10.14 |
| CHEMBL5830112 | — | — | 3 | 10.13 |
| CHEMBL6052581 | — | — | 3 | 10.13 |
| CHEMBL6027927 | — | — | 3 | 10.12 |
| CHEMBL5954405 | — | — | 3 | 10.12 |
| CHEMBL6042879 | — | — | 3 | 10.11 |
| CHEMBL5743849 | — | — | 3 | 10.11 |
| CHEMBL5286372 | — | — | 3 | 10.11 |
| CHEMBL5974502 | — | — | 3 | 10.11 |
| CHEMBL5921049 | — | — | 3 | 10.10 |
| CHEMBL6008287 | — | — | 3 | 10.09 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL5945932 | — | IC50 | = | 0.025 | nM | 10.60 |
| CHEMBL6053239 | — | IC50 | = | 0.037 | nM | 10.43 |
| CHEMBL5908882 | — | IC50 | = | 0.038 | nM | 10.42 |
| CHEMBL5784930 | — | IC50 | = | 0.039 | nM | 10.41 |
| CHEMBL5877919 | — | IC50 | = | 0.046 | nM | 10.34 |
| CHEMBL5767733 | — | IC50 | = | 0.047 | nM | 10.33 |
| CHEMBL4088737 | — | IC50 | = | 0.049 | nM | 10.31 |
| CHEMBL5798182 | — | IC50 | = | 0.05 | nM | 10.30 |
| CHEMBL5810582 | — | IC50 | = | 0.051 | nM | 10.29 |
| CHEMBL5982539 | — | IC50 | = | 0.051 | nM | 10.29 |
| CHEMBL5769925 | — | IC50 | = | 0.053 | nM | 10.28 |
| CHEMBL5857962 | — | IC50 | = | 0.054 | nM | 10.27 |
| CHEMBL6049070 | — | IC50 | = | 0.058 | nM | 10.24 |
| CHEMBL5893489 | — | IC50 | = | 0.058 | nM | 10.24 |
| CHEMBL6040479 | — | IC50 | = | 0.059 | nM | 10.23 |
| CHEMBL5826403 | — | IC50 | = | 0.065 | nM | 10.19 |
| CHEMBL5931344 | — | IC50 | = | 0.066 | nM | 10.18 |
| CHEMBL5963139 | — | IC50 | = | 0.066 | nM | 10.18 |
| CHEMBL6063394 | — | IC50 | = | 0.071 | nM | 10.15 |
| CHEMBL5820582 | — | IC50 | = | 0.073 | nM | 10.14 |
| CHEMBL6052581 | — | IC50 | = | 0.074 | nM | 10.13 |
| CHEMBL5830112 | — | IC50 | = | 0.074 | nM | 10.13 |
| CHEMBL6027927 | — | IC50 | = | 0.076 | nM | 10.12 |
| CHEMBL5954405 | — | IC50 | = | 0.075 | nM | 10.12 |
| CHEMBL6042879 | — | IC50 | = | 0.077 | nM | 10.11 |
| CHEMBL5974502 | — | IC50 | = | 0.078 | nM | 10.11 |
| CHEMBL5743849 | — | IC50 | = | 0.078 | nM | 10.11 |
| CHEMBL5286372 | — | IC50 | = | 0.078 | nM | 10.11 |
| CHEMBL5921049 | — | IC50 | = | 0.08 | nM | 10.10 |
| CHEMBL6008287 | — | IC50 | = | 0.082 | nM | 10.09 |
| CHEMBL5774354 | — | IC50 | = | 0.083 | nM | 10.08 |
| CHEMBL4059597 | — | IC50 | = | 0.083 | nM | 10.08 |
| CHEMBL5762361 | — | IC50 | = | 0.087 | nM | 10.06 |
| CHEMBL5829576 | — | IC50 | = | 0.092 | nM | 10.04 |
| CHEMBL5984909 | — | IC50 | = | 0.094 | nM | 10.03 |
| CHEMBL5786013 | — | IC50 | = | 0.098 | nM | 10.01 |
| CHEMBL5934009 | — | IC50 | = | 0.11 | nM | 9.96 |
| CHEMBL5749705 | — | IC50 | = | 0.11 | nM | 9.96 |
| CHEMBL4077454 | — | IC50 | = | 0.11 | nM | 9.96 |
| CHEMBL5852216 | — | IC50 | = | 0.11 | nM | 9.96 |
| CHEMBL6014540 | — | IC50 | = | 0.11 | nM | 9.96 |
| CHEMBL6054167 | — | IC50 | = | 0.11 | nM | 9.96 |
| CHEMBL5795610 | — | IC50 | = | 0.12 | nM | 9.92 |
| CHEMBL5859588 | — | IC50 | = | 0.12 | nM | 9.92 |
| CHEMBL5986775 | — | IC50 | = | 0.12 | nM | 9.92 |
| CHEMBL5906941 | — | IC50 | = | 0.12 | nM | 9.92 |
| CHEMBL5848305 | — | IC50 | = | 0.12 | nM | 9.92 |
| CHEMBL4071397 | — | IC50 | = | 0.12 | nM | 9.92 |
| CHEMBL5981009 | — | IC50 | = | 0.13 | nM | 9.89 |
| CHEMBL5852412 | — | IC50 | = | 0.13 | nM | 9.89 |