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Assay Detail

CHEMBL5737541

Review assay metadata, readout intent, target linkage, and publication context from the same page.

Binding
Biochemical EGFR Inhibition Assays (100 mM ATP): 5 μL of EGFR enzyme form in assay buffer (50 mM HEPES pH 7.3, 10 mM MgCl2, 1 mM EGTA, 0.01% Tween 20, 2 mM DTT) are dispensed into columns 1-23, than 5 μL of ATP and ULight-poly-GT substrate (PerkinElmer; TRF0100-M) mix in assay buffer is added to all wells (final assay concentration of the ULight-poly-GT substrate 200 nM). Each of the different EGFR enzyme form assays is available at high ATP levels (final assay concentration 100 μM). After 90 minutes incubation at room temperature 5 μL EDTA (final assay concentration 50 mM) and LANCE Eu-anti-P-Tyr (PT66) antibody (PerkinElmer, AD0069) (final assay concentration 6 nM) mix are added to stop the reaction and start the binding of the antibody. After additional 60 minutes incubation at room temperature the signal is measured in a PerkinElmer Envision HTS Multilabel Reader using the TR-FRET LANCE Ultra specs of PerkinElmer (used wavelengths: excitation 320 nm, emission 1 665 nm, emission 2 615 nm). Each plate contains 16 wells of a negative control (diluted DMSO instead of test compound; w EGFR enzyme form; column 23) and 16 wells of a positive control (diluted DMSO instead of test compound; w/o EGFR enzyme form; column 24). Negative and positive control values are used for normalization and IC50 values are calculated and analysed using a 4 parametric logistic model.
180
Total Activities
60
Compounds Tested
3
Activity Types
0
Assay Parameters

Assay Information

Assay Type Binding
Confidence 8 — Homologous single protein target
Curated By Autocuration

Target

Epidermal growth factor receptor (CHEMBL203)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Benzimidazole compounds and derivatives as EGFR inhibitors
(2021)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 60 7.32 8.52
kon 60 - -
k_off 60 - -

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL5994478 3 8.52
CHEMBL6055515 3 8.52
CHEMBL5984171 3 8.43
CHEMBL5968167 3 8.38
CHEMBL5998385 3 8.24
CHEMBL6029813 3 8.24
CHEMBL5932830 3 8.06
CHEMBL5810396 3 7.99
CHEMBL5790363 3 7.99
CHEMBL5914292 3 7.99
CHEMBL5825830 3 7.98
CHEMBL5755984 3 7.95
CHEMBL4532034 3 7.92
CHEMBL5786240 3 7.86
CHEMBL5902827 3 7.76
CHEMBL6021978 3 7.73
CHEMBL5976089 3 7.72
CHEMBL5864704 3 7.61
CHEMBL5809520 3 7.59
CHEMBL5994405 3 7.56
CHEMBL5772994 3 7.56
CHEMBL6043073 3 7.50
CHEMBL5954592 3 7.46
CHEMBL6056421 3 7.46
CHEMBL5897403 3 7.42
CHEMBL5195167 3 7.40
CHEMBL5959712 3 7.40
CHEMBL6007714 3 7.34
CHEMBL5840868 3 7.21
CHEMBL5893361 3 7.19

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL6055515 IC50 = 3.0 nM 8.52
CHEMBL5994478 IC50 = 3.0 nM 8.52
CHEMBL5984171 IC50 = 3.7 nM 8.43
CHEMBL5968167 IC50 = 4.2 nM 8.38
CHEMBL6029813 IC50 = 5.7 nM 8.24
CHEMBL5998385 IC50 = 5.8 nM 8.24
CHEMBL5932830 IC50 = 8.8 nM 8.06
CHEMBL5914292 IC50 = 10.3 nM 7.99
CHEMBL5790363 IC50 = 10.3 nM 7.99
CHEMBL5810396 IC50 = 10.3 nM 7.99
CHEMBL5825830 IC50 = 10.5 nM 7.98
CHEMBL5755984 IC50 = 11.3 nM 7.95
CHEMBL4532034 IC50 = 11.9 nM 7.92
CHEMBL5786240 IC50 = 13.7 nM 7.86
CHEMBL5902827 IC50 = 17.4 nM 7.76
CHEMBL6021978 IC50 = 18.6 nM 7.73
CHEMBL5976089 IC50 = 19.0 nM 7.72
CHEMBL5864704 IC50 = 24.3 nM 7.61
CHEMBL5809520 IC50 = 25.5 nM 7.59
CHEMBL5772994 IC50 = 27.7 nM 7.56
CHEMBL5994405 IC50 = 27.7 nM 7.56
CHEMBL6043073 IC50 = 31.6 nM 7.50
CHEMBL6056421 IC50 = 34.5 nM 7.46
CHEMBL5954592 IC50 = 34.8 nM 7.46
CHEMBL5897403 IC50 = 38.5 nM 7.42
CHEMBL5959712 IC50 = 39.6 nM 7.40
CHEMBL5195167 IC50 = 40.1 nM 7.40
CHEMBL6007714 IC50 = 45.7 nM 7.34
CHEMBL5840868 IC50 = 61.7 nM 7.21
CHEMBL5893361 IC50 = 64.6 nM 7.19
CHEMBL5960825 IC50 = 67.4 nM 7.17
CHEMBL5872035 IC50 = 76.5 nM 7.12
CHEMBL6015235 IC50 = 78.2 nM 7.11
CHEMBL5865007 IC50 = 79.3 nM 7.10
CHEMBL5891448 IC50 = 86.9 nM 7.06
CHEMBL5829030 IC50 = 91.6 nM 7.04
CHEMBL5865099 IC50 = 94.1 nM 7.03
CHEMBL5963945 IC50 = 94.6 nM 7.02
CHEMBL5911542 IC50 = 95.2 nM 7.02
CHEMBL5932930 IC50 = 102.0 nM 6.99
CHEMBL5861020 IC50 = 105.0 nM 6.98
CHEMBL5832255 IC50 = 104.0 nM 6.98
CHEMBL5985978 IC50 = 104.0 nM 6.98
CHEMBL5783119 IC50 = 111.0 nM 6.96
CHEMBL5925024 IC50 = 113.0 nM 6.95
CHEMBL5784511 IC50 = 128.0 nM 6.89
CHEMBL5834325 IC50 = 146.0 nM 6.84
CHEMBL5945119 IC50 = 171.0 nM 6.77
CHEMBL6048747 IC50 = 169.0 nM 6.77
CHEMBL5750984 IC50 = 178.0 nM 6.75