Assay Detail
Binding
CHEMBL5738682
Review assay metadata, readout intent, target linkage, and publication context from the same page.
Kat5 Assay: 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: KAT5 25 nM, 1 uM AcCoA, 2 uM H4 1-21 peptide, 30-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.
165
Total Activities
52
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
Benzisoxazole sulfonamide derivatives
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| Ki | 55 | 5.90 | 7.16 |
| kon | 55 | - | - |
| k_off | 55 | - | - |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL5993180 | — | — | 3 | 7.16 |
| CHEMBL5955544 | — | — | 3 | 6.96 |
| CHEMBL5852195 | — | — | 3 | 6.92 |
| CHEMBL5886450 | — | — | 3 | 6.89 |
| CHEMBL5927368 | — | — | 3 | 6.82 |
| CHEMBL5818085 | — | — | 3 | 6.82 |
| CHEMBL5883288 | — | — | 3 | 6.75 |
| CHEMBL5998930 | — | — | 3 | 6.75 |
| CHEMBL5812221 | — | — | 3 | 6.72 |
| CHEMBL5941367 | — | — | 3 | 6.70 |
| CHEMBL6005841 | — | — | 3 | 6.66 |
| CHEMBL5853409 | — | — | 3 | 6.58 |
| CHEMBL5959733 | — | — | 3 | 6.57 |
| CHEMBL5830107 | — | — | 3 | 6.50 |
| CHEMBL5963038 | — | — | 3 | 6.50 |
| CHEMBL5723326 | — | — | 12 | 6.43 |
| CHEMBL5792825 | — | — | 3 | 6.39 |
| CHEMBL5996948 | — | — | 3 | 6.37 |
| CHEMBL5857566 | — | — | 3 | 6.28 |
| CHEMBL5860247 | — | — | 3 | 6.15 |
| CHEMBL5925551 | — | — | 3 | 6.11 |
| CHEMBL5993009 | — | — | 3 | 6.10 |
| CHEMBL6057979 | — | — | 3 | 6.08 |
| CHEMBL5871508 | — | — | 3 | 6.01 |
| CHEMBL5924783 | — | — | 3 | 5.99 |
| CHEMBL5867166 | — | — | 3 | 5.99 |
| CHEMBL5847041 | — | — | 3 | 5.93 |
| CHEMBL5779221 | — | — | 3 | 5.84 |
| CHEMBL5980695 | — | — | 3 | 5.77 |
| CHEMBL5785798 | — | — | 3 | 5.68 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL5993180 | — | Ki | = | 70.0 | nM | 7.16 |
| CHEMBL5955544 | — | Ki | = | 110.0 | nM | 6.96 |
| CHEMBL5852195 | — | Ki | = | 120.0 | nM | 6.92 |
| CHEMBL5886450 | — | Ki | = | 130.0 | nM | 6.89 |
| CHEMBL5927368 | — | Ki | = | 150.0 | nM | 6.82 |
| CHEMBL5818085 | — | Ki | = | 150.0 | nM | 6.82 |
| CHEMBL5883288 | — | Ki | = | 180.0 | nM | 6.75 |
| CHEMBL5998930 | — | Ki | = | 180.0 | nM | 6.75 |
| CHEMBL5812221 | — | Ki | = | 190.0 | nM | 6.72 |
| CHEMBL5941367 | — | Ki | = | 200.0 | nM | 6.70 |
| CHEMBL6005841 | — | Ki | = | 220.0 | nM | 6.66 |
| CHEMBL5853409 | — | Ki | = | 260.0 | nM | 6.58 |
| CHEMBL5959733 | — | Ki | = | 270.0 | nM | 6.57 |
| CHEMBL5963038 | — | Ki | = | 320.0 | nM | 6.50 |
| CHEMBL5830107 | — | Ki | = | 320.0 | nM | 6.50 |
| CHEMBL5723326 | — | Ki | = | 370.0 | nM | 6.43 |
| CHEMBL5792825 | — | Ki | = | 410.0 | nM | 6.39 |
| CHEMBL5996948 | — | Ki | = | 430.0 | nM | 6.37 |
| CHEMBL5857566 | — | Ki | = | 530.0 | nM | 6.28 |
| CHEMBL5723326 | — | Ki | = | 570.0 | nM | 6.24 |
| CHEMBL5723326 | — | Ki | = | 620.0 | nM | 6.21 |
| CHEMBL5723326 | — | Ki | = | 610.0 | nM | 6.21 |
| CHEMBL5860247 | — | Ki | = | 710.0 | nM | 6.15 |
| CHEMBL5925551 | — | Ki | = | 770.0 | nM | 6.11 |
| CHEMBL5993009 | — | Ki | = | 800.0 | nM | 6.10 |
| CHEMBL6057979 | — | Ki | = | 830.0 | nM | 6.08 |
| CHEMBL5871508 | — | Ki | = | 980.0 | nM | 6.01 |
| CHEMBL5867166 | — | Ki | = | 1030.0 | nM | 5.99 |
| CHEMBL5924783 | — | Ki | = | 1030.0 | nM | 5.99 |
| CHEMBL5847041 | — | Ki | = | 1170.0 | nM | 5.93 |
| CHEMBL5779221 | — | Ki | = | 1440.0 | nM | 5.84 |
| CHEMBL5980695 | — | Ki | = | 1710.0 | nM | 5.77 |
| CHEMBL5785798 | — | Ki | = | 2070.0 | nM | 5.68 |
| CHEMBL5938038 | — | Ki | = | 2960.0 | nM | 5.53 |
| CHEMBL5930092 | — | Ki | = | 3050.0 | nM | 5.52 |
| CHEMBL5808891 | — | Ki | = | 3060.0 | nM | 5.51 |
| CHEMBL5759534 | — | Ki | = | 3640.0 | nM | 5.44 |
| CHEMBL5789111 | — | Ki | = | 4380.0 | nM | 5.36 |
| CHEMBL5842425 | — | Ki | = | 4530.0 | nM | 5.34 |
| CHEMBL5756543 | — | Ki | = | 4630.0 | nM | 5.33 |
| CHEMBL5907267 | — | Ki | = | 5700.0 | nM | 5.24 |
| CHEMBL5906744 | — | Ki | = | 5980.0 | nM | 5.22 |
| CHEMBL6020167 | — | Ki | = | 6220.0 | nM | 5.21 |
| CHEMBL5782689 | — | Ki | = | 8450.0 | nM | 5.07 |
| CHEMBL5766096 | — | Ki | = | 9790.0 | nM | 5.01 |
| CHEMBL5871884 | — | Ki | = | 9920.0 | nM | 5.00 |
| CHEMBL6044138 | — | Ki | = | 10800.0 | nM | 4.97 |
| CHEMBL5780277 | — | Ki | = | 11700.0 | nM | 4.93 |
| CHEMBL6008690 | — | Ki | = | 11700.0 | nM | 4.93 |
| CHEMBL6051437 | — | Ki | = | 12500.0 | nM | 4.90 |