Assay Detail
Binding
CHEMBL5738679
Review assay metadata, readout intent, target linkage, and publication context from the same page.
Kat6A Assay 2: A. Compound preparation 1. Prepare 10 mM stock solutions in 100% DMSO from solid material 2. Serial dilute 10 mM, 1 mM or 0.1 mM compound stocks 3-fold in 100% DMSO for 11-point dose responseB. Reagent preparation 1. Prepare 1× assay buffer containing 10 mM Tris HCL pH 8.0, 2.5 mM NaCl, 0.5 mM EDTA, 0.005% BSG and 0.02% Tween-20 2. Dilute Histone peptide (CPC Scientific) and AcCoA (Sigma) together in assay buffer to 2×. 3. Dilute KAT enzyme in assay buffer to 2×.C. Enzyme reaction 1. Final reaction conditions for each KAT assay in a 20 ul assay reaction volume: KAT6A 15 nM, 10 uM AcCoA, 2 uM H3 1-21 peptide, 45-minute reaction 2. Add 0.5 ul of diluted compound to the assay plate (384-well V-bottom polypropylene plates) or 0.5 ul of DMSO for control wells. 3. Add 10 ul of 2× Histone peptide/2×AcCoA mix to the assay plate. 4. Add 10 ul of 2× enzyme to the assay plate. 5. Stop the reaction after the indicated time with the addition of 2 ul of 5% formic acid 6. Each reaction was analyzed using self-assembled monolayer desorption/ionization time-of-flight mass spectrometry (Mrksich, Milan (2008) Mass Spectrometry of Self-Assembled Monolayers: A New Tool for Molecular Surface Science ACS Nano 2008 2 (1), 7-18; SAMDI Tech, Inc. (Chicago, Ill.)). 7. Area under the curve (AUC) for both substrate and product peaks was determined for KAT5 at M.W. 2561 [Substrate+H]+ and 2603 [Product+H]+ with a +/−1 Da tolerance, respectively 8. Area under the curve (AUC) for both substrate and product peaks was determined for KAT6a, KAT6B, KAT7 and KAT8 at M.W. 2723 [Substrate+H]+ and 2765 [Product+H]+ with a +/−1 Da tolerance, respectively. 9. Percent conversion to product was calculated by: AUCProduct/(AUCsubstrate+AUCProduct).D. Data analysis 1. IC50 values were determined by fitting the % conversion at each inhibitor concentration to the 4-parameter IC50 equation using Pfizer proprietary curve fitting software. 2. Ki values were determined by fitting the % conversion at each inhibitor concentration to the Morrison equation for tightbinding competitive inhibitors using Pfizer proprietary curve fitting software.
225
Total Activities
71
Compounds Tested
3
Activity Types
0
Assay Parameters
Assay Information
| Assay Type | Binding |
| Confidence | 8 — Homologous single protein target |
| Curated By | Autocuration |
Publication
Benzisoxazole sulfonamide derivatives
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| Ki | 75 | 8.49 | 9.46 |
| kon | 75 | - | - |
| k_off | 75 | - | - |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL5993180 | — | — | 3 | 9.46 |
| CHEMBL5886450 | — | — | 3 | 9.42 |
| CHEMBL5853409 | — | — | 3 | 9.40 |
| CHEMBL5852195 | — | — | 3 | 9.37 |
| CHEMBL5955544 | — | — | 3 | 9.35 |
| CHEMBL5812221 | — | — | 3 | 9.34 |
| CHEMBL5883288 | — | — | 3 | 9.31 |
| CHEMBL5959733 | — | — | 3 | 9.31 |
| CHEMBL5818085 | — | — | 3 | 9.29 |
| CHEMBL6000330 | — | — | 3 | 9.29 |
| CHEMBL5853561 | — | — | 3 | 9.23 |
| CHEMBL6057979 | — | — | 3 | 9.19 |
| CHEMBL5723326 | — | — | 12 | 9.18 |
| CHEMBL6005841 | — | — | 3 | 9.17 |
| CHEMBL5927368 | — | — | 3 | 9.06 |
| CHEMBL5941367 | — | — | 3 | 9.04 |
| CHEMBL5996948 | — | — | 3 | 8.99 |
| CHEMBL5830107 | — | — | 3 | 8.93 |
| CHEMBL5785798 | — | — | 3 | 8.91 |
| CHEMBL5857566 | — | — | 3 | 8.86 |
| CHEMBL5766096 | — | — | 3 | 8.81 |
| CHEMBL5925551 | — | — | 3 | 8.78 |
| CHEMBL5860247 | — | — | 3 | 8.77 |
| CHEMBL5847041 | — | — | 3 | 8.77 |
| CHEMBL6051437 | — | — | 3 | 8.74 |
| CHEMBL5794113 | — | — | 3 | 8.68 |
| CHEMBL5871508 | — | — | 3 | 8.63 |
| CHEMBL5938038 | — | — | 3 | 8.63 |
| CHEMBL5760697 | — | — | 3 | 8.59 |
| CHEMBL5945088 | — | — | 3 | 8.50 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL5993180 | — | Ki | = | 0.35 | nM | 9.46 |
| CHEMBL5886450 | — | Ki | = | 0.38 | nM | 9.42 |
| CHEMBL5853409 | — | Ki | = | 0.4 | nM | 9.40 |
| CHEMBL5852195 | — | Ki | = | 0.43 | nM | 9.37 |
| CHEMBL5955544 | — | Ki | = | 0.45 | nM | 9.35 |
| CHEMBL5812221 | — | Ki | = | 0.46 | nM | 9.34 |
| CHEMBL5883288 | — | Ki | = | 0.49 | nM | 9.31 |
| CHEMBL5959733 | — | Ki | = | 0.49 | nM | 9.31 |
| CHEMBL6000330 | — | Ki | = | 0.51 | nM | 9.29 |
| CHEMBL5818085 | — | Ki | = | 0.51 | nM | 9.29 |
| CHEMBL5853561 | — | Ki | = | 0.59 | nM | 9.23 |
| CHEMBL6057979 | — | Ki | = | 0.64 | nM | 9.19 |
| CHEMBL5723326 | — | Ki | = | 0.66 | nM | 9.18 |
| CHEMBL6005841 | — | Ki | = | 0.68 | nM | 9.17 |
| CHEMBL5723326 | — | Ki | = | 0.75 | nM | 9.12 |
| CHEMBL5723326 | — | Ki | = | 0.8 | nM | 9.10 |
| CHEMBL5723326 | — | Ki | = | 0.83 | nM | 9.08 |
| CHEMBL5927368 | — | Ki | = | 0.87 | nM | 9.06 |
| CHEMBL5941367 | — | Ki | = | 0.92 | nM | 9.04 |
| CHEMBL5996948 | — | Ki | = | 1.02 | nM | 8.99 |
| CHEMBL5830107 | — | Ki | = | 1.17 | nM | 8.93 |
| CHEMBL5785798 | — | Ki | = | 1.24 | nM | 8.91 |
| CHEMBL5857566 | — | Ki | = | 1.38 | nM | 8.86 |
| CHEMBL5766096 | — | Ki | = | 1.54 | nM | 8.81 |
| CHEMBL5925551 | — | Ki | = | 1.67 | nM | 8.78 |
| CHEMBL5860247 | — | Ki | = | 1.7 | nM | 8.77 |
| CHEMBL5847041 | — | Ki | = | 1.7 | nM | 8.77 |
| CHEMBL6051437 | — | Ki | = | 1.83 | nM | 8.74 |
| CHEMBL5794113 | — | Ki | = | 2.1 | nM | 8.68 |
| CHEMBL5938038 | — | Ki | = | 2.36 | nM | 8.63 |
| CHEMBL5871508 | — | Ki | = | 2.32 | nM | 8.63 |
| CHEMBL5760697 | — | Ki | = | 2.6 | nM | 8.59 |
| CHEMBL5945088 | — | Ki | = | 3.16 | nM | 8.50 |
| CHEMBL5980695 | — | Ki | = | 3.23 | nM | 8.49 |
| CHEMBL5792825 | — | Ki | = | 3.27 | nM | 8.48 |
| CHEMBL5789111 | — | Ki | = | 3.62 | nM | 8.44 |
| CHEMBL5814838 | — | Ki | = | 3.9 | nM | 8.41 |
| CHEMBL5804751 | — | Ki | = | 3.89 | nM | 8.41 |
| CHEMBL5827996 | — | Ki | = | 4.14 | nM | 8.38 |
| CHEMBL5808891 | — | Ki | = | 4.44 | nM | 8.35 |
| CHEMBL6005999 | — | Ki | = | 4.65 | nM | 8.33 |
| CHEMBL5929534 | — | Ki | = | 4.64 | nM | 8.33 |
| CHEMBL5779221 | — | Ki | = | 4.98 | nM | 8.30 |
| CHEMBL5866271 | — | Ki | = | 5.23 | nM | 8.28 |
| CHEMBL6032068 | — | Ki | = | 5.32 | nM | 8.27 |
| CHEMBL5896279 | — | Ki | = | 5.57 | nM | 8.25 |
| CHEMBL5756543 | — | Ki | = | 5.61 | nM | 8.25 |
| CHEMBL5871884 | — | Ki | = | 5.73 | nM | 8.24 |
| CHEMBL6000207 | — | Ki | = | 6.25 | nM | 8.20 |
| CHEMBL5976778 | — | Ki | = | 6.28 | nM | 8.20 |