Assay Detail
Binding
CHEMBL5733237
Review assay metadata, readout intent, target linkage, and publication context from the same page.
ASV & NPG EGFR Exon 20 Insertion Mutations: A compound's ability in selectively inhibiting EGFR exon 20 insertion mutations can be assessed using Ba/F3 cells, a murine pro-B cell line, which have been transduced with EGFR exon 20 insertions. An expression vector, pLVX-IRES puro (Clontech) coding for human EGFR exon 20 insertions NPG (H773_V774insNPG) or ASV (V769_D770insASV), was transfected into HEK293 cells by the Trans-Lentiviral ORF Packaging System (Thermo Scientific), to produce virus encoding EGFR exon 20 insertions. Ba/F3 (DSMZ) cells maintained in RPMI 1640 medium supplemented with 10% fetal bovine serum, 200 μM L-glutamine/200 μg/mL penicillin/200 μg/mL streptomycin (Life Technology) and 10 ng/mL IL-3 (R&D system), were infected by EGFR Exon20 virus and subsequently selected by puromycin (Life Technology) selection and IL-3 depletion. Ba/F3 cells expressing EGFR exon 20 insertions (named Ba/F3-EGFR-Exon20-NPG or Ba/F3-EGFR-Exon20-ASV) can proliferate in the absence of IL-3. The anti-proliferative activity of compounds were determined as follows: BaF3-EGFR-Exon20 cells (NPG or ASV) seeded in 96 well plates (2500 cells/well) were treated with test compound (dissolved in DMSO) at a series of concentrations (4-fold dilution, top concentration: 10,000 nM). The plates were incubated for 72 h in a 37° C. incubator with 5% CO2, and the number of viable cells in each well were measured indirectly by CellTiter 96® Aqueous One Solution Cell Proliferation Assay (Promega) This assay is a colorimetric method for determining the number of viable cells through measurement of their metabolic activity by detection of enzymatic conversion of tetrazolium salts into blue formazan derivatives. Reagent (20 μL) was added into each well, and the plates were returned to the incubator for 2 h. The absorbance in each well was then measured at 490 nm using an Envision plate reader (Perkin Elmer). IC50 values were calculated by determining the concentration of compound required to decrease the MTS signal by 50% comparing to the DMSO control in best-fit curves using Microsoft XLfit software or Accelrys Pipeline Pilot.
498
Total Activities
99
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
Heteroaryl compounds for kinase inhibition
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| IC50 | 166 | 6.44 | 6.52 |
| kon | 166 | - | - |
| k_off | 166 | - | - |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL5787865 | — | — | 6 | 6.52 |
| CHEMBL5870314 | — | — | 3 | 6.52 |
| CHEMBL5780612 | — | — | 3 | 6.52 |
| CHEMBL5960275 | — | — | 3 | 6.52 |
| CHEMBL5930490 | — | — | 6 | 6.52 |
| CHEMBL5820070 | — | — | 6 | 6.52 |
| CHEMBL5936272 | — | — | 6 | 6.52 |
| CHEMBL5820028 | — | — | 6 | 6.52 |
| CHEMBL3353412 | — | — | 6 | 6.52 |
| CHEMBL5773016 | — | — | 6 | 6.52 |
| CHEMBL6058940 | — | — | 6 | 6.52 |
| CHEMBL6009791 | — | — | 6 | 6.52 |
| CHEMBL6029316 | — | — | 6 | 6.52 |
| CHEMBL5749382 | — | — | 3 | 6.52 |
| CHEMBL6034048 | — | — | 6 | 6.52 |
| CHEMBL5978253 | — | — | 3 | 6.52 |
| CHEMBL5949541 | — | — | 6 | 6.52 |
| CHEMBL5905455 | — | — | 3 | 6.52 |
| CHEMBL5804325 | — | — | 6 | 6.52 |
| CHEMBL5978450 | — | — | 3 | 6.52 |
| CHEMBL5802671 | — | — | 6 | 6.52 |
| CHEMBL5834572 | — | — | 6 | 6.52 |
| CHEMBL6022292 | — | — | 3 | 6.52 |
| CHEMBL5752500 | — | — | 3 | 6.52 |
| CHEMBL3353405 | — | — | 3 | 6.52 |
| CHEMBL3353410 | OSIMERTINIB | 4.0 | 6 | 6.52 |
| CHEMBL5854207 | — | — | 6 | 6.52 |
| CHEMBL5780056 | — | — | 6 | 6.52 |
| CHEMBL5828176 | — | — | 3 | 6.52 |
| CHEMBL5844856 | — | — | 6 | 6.52 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL5820028 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5949541 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5752500 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL6009791 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5804325 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5978450 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL6022292 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL6034048 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5978253 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL3353405 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5787865 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5773016 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5870116 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5787865 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL3353412 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5780612 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5802671 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5960275 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5905455 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5930490 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5936272 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL3353410 | OSIMERTINIB | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5854207 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5818238 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5870314 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5780056 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5844856 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL6058583 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL6029316 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL6058940 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5749382 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5820070 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5828176 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL6009791 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5834572 | — | IC50 | = | 300.0 | nM | 6.52 |
| CHEMBL5824145 | — | IC50 | = | 750.0 | nM | 6.12 |
| CHEMBL5907594 | — | IC50 | = | 750.0 | nM | 6.12 |
| CHEMBL5838931 | — | IC50 | = | 750.0 | nM | 6.12 |
| CHEMBL5848181 | — | IC50 | = | 750.0 | nM | 6.12 |
| CHEMBL5802671 | — | IC50 | = | 750.0 | nM | 6.12 |
| CHEMBL5942093 | — | IC50 | = | 750.0 | nM | 6.12 |
| CHEMBL5766497 | — | IC50 | = | 750.0 | nM | 6.12 |
| CHEMBL5976731 | — | IC50 | = | 750.0 | nM | 6.12 |
| CHEMBL2426277 | — | IC50 | = | 750.0 | nM | 6.12 |
| CHEMBL3353404 | — | kon | = | - | — | - |
| CHEMBL6034073 | — | IC50 | < | 100.0 | nM | - |
| CHEMBL6034073 | — | kon | = | - | — | - |
| CHEMBL6034073 | — | k_off | = | - | s-1 | - |
| CHEMBL6034073 | — | IC50 | < | 100.0 | nM | - |
| CHEMBL5854576 | — | kon | = | - | — | - |