Assay Detail
Binding
CHEMBL5732440
Review assay metadata, readout intent, target linkage, and publication context from the same page.
Chromatin Immunoprecipitation (CHIP): ChIP assay was performed using EZ ChIP Kit (Millipore, Billerica, Mass., USA) as per manufacturer's instruction. Briefly, after crosslinking with formaldehyde (1%), cell were lysed using SDS Lysis Buffer containing Protease Inhibitor Cocktail and nuclear extract were obtained. Then, crosslinked DNA was sheared using sonication; sheared DNA size was checked on agarose gel and ranged, in all cases, between 300-500 bp. An amount of 1/20 of the shared chromatin was kept as an input. Sheared chromatin was incubated with rabbit polyclonal Phospho-Stat5 (Tyr694) Antibody (Cell Signaling, Danvers, Mass., USA) at dilution 1:50 or rabbit polyclonal IgG at appropriate concentration. Incubation was performed at 4° C. overnight. The antibody/chromatin complex was precipitated using Protein G Agarose beads. After elution, the crosslink of protein/DNA was reversed, and DNA was purified using Spin Columns. PCR was performed using platinum Taq (Invitrogen Canada, Burlington, ON, Canada). Two sets of primers were designed to amplify two DNA segments that contain STAT5-binding sites located 1672 bp and 428 bp upstream the start sites of C-Myc and Cyclin-D1, respectively. The STAT5-binding site in the C-MYC promoter is characterized by a 4N spacer and has the sequence (ttcccccgaa), whereas the one in the Cyclin D1 promoter is characterized by a 3N spacer and has the sequence (ttcttggaa).
279
Total Activities
86
Compounds Tested
4
Activity Types
0
Assay Parameters
Assay Information
| Assay Type | Binding |
| Confidence | 8 — Homologous single protein target |
| Curated By | Autocuration |
Target
Signal transducer and activator of transcription 3 (CHEMBL4026) ↗| Type | SINGLE PROTEIN |
| Organism | Homo sapiens |
Publication
Substituted 2-hydroxy-4-(2-(phenylsulfonamido)acetamido)benzoic acid analogs as inhibitors of STAT protein
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| kon | 93 | - | - |
| k_off | 93 | - | - |
| Ki | 50 | 5.11 | 5.75 |
| IC50 | 43 | 4.26 | 5.17 |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL2431993 | — | — | 3 | 5.75 |
| CHEMBL2431970 | — | — | 3 | 5.60 |
| CHEMBL2431942 | — | — | 3 | 5.60 |
| CHEMBL2431990 | — | — | 3 | 5.58 |
| CHEMBL2431991 | — | — | 3 | 5.58 |
| CHEMBL2431964 | — | — | 3 | 5.52 |
| CHEMBL2431965 | — | — | 3 | 5.51 |
| CHEMBL2431966 | — | — | 3 | 5.44 |
| CHEMBL2431945 | — | — | 3 | 5.42 |
| CHEMBL2431962 | — | — | 3 | 5.42 |
| CHEMBL2431967 | — | — | 3 | 5.40 |
| CHEMBL5974948 | — | — | 3 | 5.37 |
| CHEMBL2431961 | — | — | 3 | 5.36 |
| CHEMBL2431941 | — | — | 3 | 5.35 |
| CHEMBL2431995 | — | — | 3 | 5.34 |
| CHEMBL2431944 | — | — | 3 | 5.32 |
| CHEMBL2431969 | — | — | 3 | 5.30 |
| CHEMBL2431994 | — | — | 3 | 5.29 |
| CHEMBL2431960 | — | — | 3 | 5.28 |
| CHEMBL2431943 | — | — | 3 | 5.26 |
| CHEMBL2431971 | — | — | 3 | 5.26 |
| CHEMBL2431947 | — | — | 3 | 5.24 |
| CHEMBL2431968 | — | — | 3 | 5.23 |
| CHEMBL2431973 | — | — | 3 | 5.21 |
| CHEMBL5952999 | — | — | 3 | 5.21 |
| CHEMBL1829876 | — | — | 6 | 5.17 |
| CHEMBL2431989 | — | — | 3 | 5.14 |
| CHEMBL2431939 | — | — | 3 | 5.11 |
| CHEMBL2431963 | — | — | 3 | 5.10 |
| CHEMBL2431986 | — | — | 3 | 5.08 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL2431993 | — | Ki | = | 1800.0 | nM | 5.75 |
| CHEMBL2431970 | — | Ki | = | 2500.0 | nM | 5.60 |
| CHEMBL2431942 | — | Ki | = | 2500.0 | nM | 5.60 |
| CHEMBL2431991 | — | Ki | = | 2600.0 | nM | 5.58 |
| CHEMBL2431990 | — | Ki | = | 2600.0 | nM | 5.58 |
| CHEMBL2431964 | — | Ki | = | 3000.0 | nM | 5.52 |
| CHEMBL2431965 | — | Ki | = | 3100.0 | nM | 5.51 |
| CHEMBL2431966 | — | Ki | = | 3600.0 | nM | 5.44 |
| CHEMBL2431962 | — | Ki | = | 3800.0 | nM | 5.42 |
| CHEMBL2431945 | — | Ki | = | 3800.0 | nM | 5.42 |
| CHEMBL2431967 | — | Ki | = | 4000.0 | nM | 5.40 |
| CHEMBL5974948 | — | Ki | = | 4300.0 | nM | 5.37 |
| CHEMBL2431961 | — | Ki | = | 4400.0 | nM | 5.36 |
| CHEMBL2431941 | — | Ki | = | 4500.0 | nM | 5.35 |
| CHEMBL2431995 | — | Ki | = | 4600.0 | nM | 5.34 |
| CHEMBL2431944 | — | Ki | = | 4800.0 | nM | 5.32 |
| CHEMBL2431969 | — | Ki | = | 5000.0 | nM | 5.30 |
| CHEMBL2431994 | — | Ki | = | 5100.0 | nM | 5.29 |
| CHEMBL2431960 | — | Ki | = | 5300.0 | nM | 5.28 |
| CHEMBL2431971 | — | Ki | = | 5500.0 | nM | 5.26 |
| CHEMBL2431943 | — | Ki | = | 5500.0 | nM | 5.26 |
| CHEMBL2431947 | — | Ki | = | 5800.0 | nM | 5.24 |
| CHEMBL2431968 | — | Ki | = | 5900.0 | nM | 5.23 |
| CHEMBL2431973 | — | Ki | = | 6200.0 | nM | 5.21 |
| CHEMBL5952999 | — | Ki | = | 6200.0 | nM | 5.21 |
| CHEMBL1829876 | — | IC50 | = | 6800.0 | nM | 5.17 |
| CHEMBL2431989 | — | Ki | = | 7200.0 | nM | 5.14 |
| CHEMBL2431939 | — | Ki | = | 7700.0 | nM | 5.11 |
| CHEMBL2431963 | — | Ki | = | 7900.0 | nM | 5.10 |
| CHEMBL2431986 | — | Ki | = | 8300.0 | nM | 5.08 |
| CHEMBL2431985 | — | Ki | = | 8900.0 | nM | 5.05 |
| CHEMBL6031851 | — | Ki | = | 9200.0 | nM | 5.04 |
| CHEMBL2429889 | — | Ki | = | 11100.0 | nM | 4.96 |
| CHEMBL2431978 | — | Ki | = | 11300.0 | nM | 4.95 |
| CHEMBL2431940 | — | Ki | = | 11400.0 | nM | 4.94 |
| CHEMBL2431972 | — | Ki | = | 12000.0 | nM | 4.92 |
| CHEMBL1829876 | — | Ki | = | 13000.0 | nM | 4.89 |
| CHEMBL2023986 | — | Ki | = | 16700.0 | nM | 4.78 |
| CHEMBL5866289 | — | Ki | = | 18700.0 | nM | 4.73 |
| CHEMBL2023987 | — | Ki | = | 21500.0 | nM | 4.67 |
| CHEMBL5845912 | — | Ki | = | 21600.0 | nM | 4.67 |
| CHEMBL2431975 | — | Ki | = | 22600.0 | nM | 4.65 |
| CHEMBL2431977 | — | Ki | = | 23300.0 | nM | 4.63 |
| CHEMBL1829864 | — | IC50 | = | 24600.0 | nM | 4.61 |
| CHEMBL2431976 | — | Ki | = | 25400.0 | nM | 4.59 |
| CHEMBL1829871 | — | Ki | = | 26500.0 | nM | 4.58 |
| CHEMBL2431987 | — | Ki | = | 27900.0 | nM | 4.55 |
| CHEMBL1829872 | — | IC50 | = | 28800.0 | nM | 4.54 |
| CHEMBL1829871 | — | IC50 | = | 28800.0 | nM | 4.54 |
| CHEMBL5808733 | — | Ki | = | 33000.0 | nM | 4.48 |