Assay Detail
Binding
CHEMBL5738684
Review assay metadata, readout intent, target linkage, and publication context from the same page.
Kat8 Assay: TBDA. 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: KAT8 15 nM, 1 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.
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 | 5.67 | 7.00 |
| kon | 55 | - | - |
| k_off | 55 | - | - |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL5886450 | — | — | 3 | 7.00 |
| CHEMBL5852195 | — | — | 3 | 6.96 |
| CHEMBL5812221 | — | — | 3 | 6.80 |
| CHEMBL5993180 | — | — | 3 | 6.77 |
| CHEMBL5818085 | — | — | 3 | 6.66 |
| CHEMBL5955544 | — | — | 3 | 6.62 |
| CHEMBL5853409 | — | — | 3 | 6.46 |
| CHEMBL5959733 | — | — | 3 | 6.38 |
| CHEMBL5927368 | — | — | 3 | 6.33 |
| CHEMBL5996948 | — | — | 3 | 6.33 |
| CHEMBL5723326 | — | — | 12 | 6.30 |
| CHEMBL6005841 | — | — | 3 | 6.28 |
| CHEMBL5941367 | — | — | 3 | 6.27 |
| CHEMBL5883288 | — | — | 3 | 6.25 |
| CHEMBL5857566 | — | — | 3 | 6.18 |
| CHEMBL5924783 | — | — | 3 | 6.08 |
| CHEMBL5963038 | — | — | 3 | 6.05 |
| CHEMBL5993009 | — | — | 3 | 6.02 |
| CHEMBL5998930 | — | — | 3 | 6.00 |
| CHEMBL5847041 | — | — | 3 | 5.97 |
| CHEMBL5925551 | — | — | 3 | 5.96 |
| CHEMBL5830107 | — | — | 3 | 5.84 |
| CHEMBL5867166 | — | — | 3 | 5.82 |
| CHEMBL6057979 | — | — | 3 | 5.76 |
| CHEMBL5785798 | — | — | 3 | 5.71 |
| CHEMBL5980695 | — | — | 3 | 5.54 |
| CHEMBL6007870 | — | — | 3 | 5.49 |
| CHEMBL5779221 | — | — | 3 | 5.49 |
| CHEMBL5930092 | — | — | 3 | 5.49 |
| CHEMBL5759534 | — | — | 3 | 5.49 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL5886450 | — | Ki | = | 100.0 | nM | 7.00 |
| CHEMBL5852195 | — | Ki | = | 110.0 | nM | 6.96 |
| CHEMBL5812221 | — | Ki | = | 160.0 | nM | 6.80 |
| CHEMBL5993180 | — | Ki | = | 170.0 | nM | 6.77 |
| CHEMBL5818085 | — | Ki | = | 220.0 | nM | 6.66 |
| CHEMBL5955544 | — | Ki | = | 240.0 | nM | 6.62 |
| CHEMBL5853409 | — | Ki | = | 350.0 | nM | 6.46 |
| CHEMBL5959733 | — | Ki | = | 420.0 | nM | 6.38 |
| CHEMBL5927368 | — | Ki | = | 470.0 | nM | 6.33 |
| CHEMBL5996948 | — | Ki | = | 470.0 | nM | 6.33 |
| CHEMBL5723326 | — | Ki | = | 500.0 | nM | 6.30 |
| CHEMBL6005841 | — | Ki | = | 530.0 | nM | 6.28 |
| CHEMBL5941367 | — | Ki | = | 540.0 | nM | 6.27 |
| CHEMBL5883288 | — | Ki | = | 560.0 | nM | 6.25 |
| CHEMBL5857566 | — | Ki | = | 660.0 | nM | 6.18 |
| CHEMBL5723326 | — | Ki | = | 660.0 | nM | 6.18 |
| CHEMBL5723326 | — | Ki | = | 730.0 | nM | 6.14 |
| CHEMBL5723326 | — | Ki | = | 820.0 | nM | 6.09 |
| CHEMBL5924783 | — | Ki | = | 830.0 | nM | 6.08 |
| CHEMBL5963038 | — | Ki | = | 900.0 | nM | 6.05 |
| CHEMBL5993009 | — | Ki | = | 960.0 | nM | 6.02 |
| CHEMBL5998930 | — | Ki | = | 1010.0 | nM | 6.00 |
| CHEMBL5847041 | — | Ki | = | 1070.0 | nM | 5.97 |
| CHEMBL5925551 | — | Ki | = | 1110.0 | nM | 5.96 |
| CHEMBL5830107 | — | Ki | = | 1440.0 | nM | 5.84 |
| CHEMBL5867166 | — | Ki | = | 1510.0 | nM | 5.82 |
| CHEMBL6057979 | — | Ki | = | 1750.0 | nM | 5.76 |
| CHEMBL5785798 | — | Ki | = | 1970.0 | nM | 5.71 |
| CHEMBL5980695 | — | Ki | = | 2870.0 | nM | 5.54 |
| CHEMBL5779221 | — | Ki | = | 3260.0 | nM | 5.49 |
| CHEMBL5792825 | — | Ki | = | 3260.0 | nM | 5.49 |
| CHEMBL6007870 | — | Ki | = | 3260.0 | nM | 5.49 |
| CHEMBL5759534 | — | Ki | = | 3260.0 | nM | 5.49 |
| CHEMBL5896279 | — | Ki | = | 3260.0 | nM | 5.49 |
| CHEMBL5930092 | — | Ki | = | 3260.0 | nM | 5.49 |
| CHEMBL5860247 | — | Ki | = | 3260.0 | nM | 5.49 |
| CHEMBL5871508 | — | Ki | = | 3320.0 | nM | 5.48 |
| CHEMBL5842425 | — | Ki | = | 4460.0 | nM | 5.35 |
| CHEMBL6020167 | — | Ki | = | 4950.0 | nM | 5.30 |
| CHEMBL6051437 | — | Ki | = | 12500.0 | nM | 4.90 |
| CHEMBL5782689 | — | Ki | = | 12500.0 | nM | 4.90 |
| CHEMBL5789111 | — | Ki | = | 12500.0 | nM | 4.90 |
| CHEMBL5871884 | — | Ki | = | 12500.0 | nM | 4.90 |
| CHEMBL5756543 | — | Ki | = | 12500.0 | nM | 4.90 |
| CHEMBL5907267 | — | Ki | = | 12500.0 | nM | 4.90 |
| CHEMBL5808891 | — | Ki | = | 12500.0 | nM | 4.90 |
| CHEMBL6008690 | — | Ki | = | 12500.0 | nM | 4.90 |
| CHEMBL5938038 | — | Ki | = | 12500.0 | nM | 4.90 |
| CHEMBL5780277 | — | Ki | = | 17400.0 | nM | 4.76 |
| CHEMBL5766096 | — | Ki | = | 17400.0 | nM | 4.76 |