WZ4002 [Ligand Id: 9176] activity data from GtoPdb and ChEMBL
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| ChEMBL ligand: CHEMBL1229592 |
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| DB | Assay description | Assay Type | Standard value | Standard parameter | Original value | Original units | Original parameter | Reference |
|---|---|---|---|---|---|---|---|---|
| ALK receptor tyrosine kinase/ALK tyrosine kinase receptor in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4247] [GtoPdb: 1839] [UniProtKB: Q9UM73] | ||||||||
| ChEMBL | Binding affinity of ALK (unknown origin) assessed as dissociation constant | B | 6.52 | pKd | 300 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| ChEMBL | Inhibition of GST-tagged human ALK cytoplasmic domain (1058 to 1620 residues) expressed in baculovirus expression system using Poly G:T (4:1) as substrate after 30 mins by Z-LYTE assay | B | 6 | pIC50 | 990 | nM | IC50 | Eur J Med Chem (2017) 136: 497-510 [PMID:28528303] |
| cyclin G associated kinase/Cyclin-G-associated kinase in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4355] [GtoPdb: 2027] [UniProtKB: O14976] | ||||||||
| ChEMBL | Binding affinity to GAK (unknown origin) assessed as dissociation constant | B | 7.04 | pKd | 91 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| BMX non-receptor tyrosine kinase/Cytoplasmic tyrosine-protein kinase BMX in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL3834] [GtoPdb: 1942] [UniProtKB: P51813] | ||||||||
| ChEMBL | Inhibition of TEL fused BMX (unknown origin) transformed in mouse BaF3 cells | B | 7.32 | pIC50 | 48 | nM | IC50 | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| cathepsin C/Dipeptidyl peptidase 1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2252] [GtoPdb: 2344] [UniProtKB: P53634] | ||||||||
| ChEMBL | Inhibition of CatL-activated recombinant human C-terminal His10-tagged cathepsin C (25 to 463 residues) expressed in mouse myeloma cells using Gly-Phe-AFC as substrate preincubated for 3 hrs followed by substrate addition and measured after 60 mins | B | 5.68 | pIC50 | 2100 | nM | IC50 | J Med Chem (2019) 62: 5901-5919 [PMID:31145622] |
| ChEMBL | Inhibition of cathepsin C (unknown origin) | B | 5.68 | pIC50 | 2100 | nM | IC50 | J Med Chem (2019) 62: 5901-5919 [PMID:31145622] |
| eukaryotic translation initiation factor 2 alpha kinase 4/eIF-2-alpha kinase GCN2 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL5358] [GtoPdb: 2018] [UniProtKB: Q9P2K8] | ||||||||
| ChEMBL | Binding affinity to GCN2 (unknown origin) assessed as dissociation constant | B | 6.85 | pKd | 140 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| epidermal growth factor receptor/Epidermal growth factor receptor in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL203] [GtoPdb: 1797] [UniProtKB: P00533] | ||||||||
| ChEMBL | Binding affinity to recombinant EGFR L858R/T790M mutant | B | 6.89 | pKd | 130 | nM | Kd | J Med Chem (2012) 55: 6243-6262 [PMID:22621397] |
| ChEMBL | Binding affinity to EGFR (unknown origin) assessed as dissociation constant | B | 7.34 | pKd | 46 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| ChEMBL | EGFR in vitro assay: Kinase-tagged T7 phage strains were prepared in an E. coli host derived from the BL21 strain. E. coli were grown to log-phase and infected with T7 phage and incubated with shaking at 32° C. until lysis. The lysates were centrifuged and filtered to remove cell debris. The remaining kinases were produced in HEK-293 cells and subsequently tagged with DNA for qPCR detection. Streptavidin-coated magnetic beads were treated with biotinylated small molecule ligands for 30 minutes at room temperature to generate affinity resins for kinase assays. The liganded beads were blocked with excess biotin and washed with blocking buffer (SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT) to remove unbound ligand and to reduce non-specific binding. Binding reactions were assembled by combining kinases, liganded affinity beads, and test compounds in 1× binding buffer (20% SeaBlock, 0.17×PBS, 0.05% Tween 20, 6 mM DTT). All reactions were performed in polystyrene 96-well plates in a final volume of 0.135 ml. The assay plates were incubated at room temperature with shaking for 1 hour and the affinity beads were washed with wash buffer (1×PBS, 0.05% Tween 20). The beads were then re-suspended in elution buffer (1× PBS, 0.05% Tween 20, 0.5 μM non-biotinylated affinity ligand) and incubated at room temperature with shaking for 30 minutes. The kinase concentration in the eluates was measured by qPCR. | B | 7.34 | pKd | 46 | nM | Kd | US-10167264-B2. Substituted pyrimidines useful as EGFR-T790M kinase inhibitors (2019) |
| ChEMBL | Binding affinity to EGFR | B | 7.34 | pKd | 46 | nM | Kd | J Med Chem (2012) 55: 2711-2723 [PMID:22339342] |
| GtoPdb | - | - | 7.34 | pKd | 46 | nM | Kd | Nature (2009) 462: 1070-4 [PMID:20033049] |
| ChEMBL | Binding affinity to EGFR L858R mutant (unknown origin) assessed as dissociation constant | B | 7.54 | pKd | 29 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| ChEMBL | Binding affinity to EGFR L858R mutant | B | 7.54 | pKd | 29 | nM | Kd | J Med Chem (2012) 55: 2711-2723 [PMID:22339342] |
| ChEMBL | Binding affinity to EGFR del S752_I759 mutant (unknown origin) assessed as dissociation constant | B | 7.55 | pKd | 28 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| ChEMBL | Binding affinity to EGFR del L747_E749,A750P mutant (unknown origin) assessed as dissociation constant | B | 7.85 | pKd | 14 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| ChEMBL | EGFR in vitro assay: Kinase-tagged T7 phage strains were prepared in an E. coli host derived from the BL21 strain. E. coli were grown to log-phase and infected with T7 phage and incubated with shaking at 32° C. until lysis. The lysates were centrifuged and filtered to remove cell debris. The remaining kinases were produced in HEK-293 cells and subsequently tagged with DNA for qPCR detection. Streptavidin-coated magnetic beads were treated with biotinylated small molecule ligands for 30 minutes at room temperature to generate affinity resins for kinase assays. The liganded beads were blocked with excess biotin and washed with blocking buffer (SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT) to remove unbound ligand and to reduce non-specific binding. Binding reactions were assembled by combining kinases, liganded affinity beads, and test compounds in 1× binding buffer (20% SeaBlock, 0.17×PBS, 0.05% Tween 20, 6 mM DTT). All reactions were performed in polystyrene 96-well plates in a final volume of 0.135 ml. The assay plates were incubated at room temperature with shaking for 1 hour and the affinity beads were washed with wash buffer (1×PBS, 0.05% Tween 20). The beads were then re-suspended in elution buffer (1× PBS, 0.05% Tween 20, 0.5 μM non-biotinylated affinity ligand) and incubated at room temperature with shaking for 30 minutes. The kinase concentration in the eluates was measured by qPCR. | B | 7.92 | pKd | 12 | nM | Kd | US-10167264-B2. Substituted pyrimidines useful as EGFR-T790M kinase inhibitors (2019) |
| ChEMBL | Binding affinity to EGFR del E746_A750 mutant (unknown origin) assessed as dissociation constant | B | 7.92 | pKd | 12 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| ChEMBL | Binding affinity to partial length human EGFR L858R/T790M double mutant expressed in mammalian system by KINOMEscan assay | B | 9.52 | pKd | 0.3 | nM | Kd | J Med Chem (2016) 59: 6580-6594 [PMID:26882288] |
| ChEMBL | Inhibition of EGFR in human NCI-H460 cells assessed as reduction in cell viability after 72 hrs by MTT assay | B | 4.74 | pIC50 | 18210 | nM | IC50 | Bioorg Med Chem Lett (2017) 27: 4832-4837 [PMID:28974338] |
| ChEMBL | Inhibition of EGFR E746_A750 deletion/T790M mutant in human growth-resistant PC9 cells assessed as reduction in cell viability after 72 hrs by MTT assay | B | 5.07 | pIC50 | 8600 | nM | IC50 | Bioorg Med Chem Lett (2017) 27: 4832-4837 [PMID:28974338] |
| ChEMBL | Inhibition of wildtype EGFR (unknown origin) transformed in mouse BaF3 cells | B | 5.12 | pIC50 | 7560 | nM | IC50 | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| ChEMBL | Inhibition of EGFR L858R/T790M mutant (unknown origin) expressed in baculovirus expression system by ELISA | B | 5.15 | pIC50 | 7000 | nM | IC50 | Eur J Med Chem (2017) 140: 510-527 [PMID:28987609] |
| ChEMBL | Inhibition of human EGFR T790M/C797S/L858R triple mutant preincubated for 20 mins followed by [33P]ATP addition measured after 2 hrs | B | 5.69 | pIC50 | 2053 | nM | IC50 | J Med Chem (2017) 60: 5613-5637 [PMID:28603991] |
| ChEMBL | Inhibition of recombinant human N-terminally GST-tagged EGFR L858R/T790M/C797S triple mutant expressed in baculovirus in Sf9 insect cells preincubated for 20 mins followed by addition of [33P]-ATP measured after 2 hrs by filter-binding method | B | 5.69 | pIC50 | 2050 | nM | IC50 | J Med Chem (2017) 60: 4636-4656 [PMID:28482151] |
| ChEMBL | Inhibition of wild type EGFR phosphorylation in human LoVo cells after 2 hrs by fluorescence assay | B | 5.93 | pIC50 | 1180 | nM | IC50 | J Med Chem (2013) 56: 7025-7048 [PMID:23930994] |
| ChEMBL | Inhibition of wild-type EGFR phosphorylation in human A431 cells preincubated for 1 hr followed by EGF stimulation measured after 45 mins by ELISA | B | 6.15 | pIC50 | 710 | nM | IC50 | Bioorg Med Chem Lett (2018) 28: 1257-1261 [PMID:29534926] |
| ChEMBL | Inhibition of wild type GST-tagged human EGFR cytoplasmic domain (668 to 1210 residues) expressed in baculovirus expression system using Poly G:T (4:1) as substrate after 30 mins by Z-LYTE assay | B | 6.92 | pIC50 | 120 | nM | IC50 | Eur J Med Chem (2017) 136: 497-510 [PMID:28528303] |
| ChEMBL | Inhibition of EGFR E746_A750 deletion mutant in human PC9 cells assessed as reduction in cell viability after 72 hrs by MTT assay | B | 6.96 | pIC50 | 110 | nM | IC50 | Bioorg Med Chem Lett (2017) 27: 4832-4837 [PMID:28974338] |
| ChEMBL | Inhibition of wild type recombinant human EGFR preincubated for 10 mins followed by FAM-labeled peptide/ATP addition measured after 1 hr by mobility shift assay | B | 7.1 | pIC50 | 79 | nM | IC50 | Eur J Med Chem (2015) 104: 115-126 [PMID:26451770] |
| ChEMBL | Inhibition of EGFR L858R/T790M mutant phosphorylation in human NCI-H1975 cells after 1 hr by ELISA | B | 7.34 | pIC50 | 46 | nM | IC50 | Bioorg Med Chem Lett (2018) 28: 1257-1261 [PMID:29534926] |
| ChEMBL | Inhibition of EGFR exon 19 deletion activating mutant phosphorylation in human PC9 cells after 2 hrs by fluorescence assay | B | 7.36 | pIC50 | 44 | nM | IC50 | J Med Chem (2013) 56: 7025-7048 [PMID:23930994] |
| ChEMBL | Inhibition of wild-type EGFR (unknown origin) expressed in baculovirus expression system after 1 hr by ELISA | B | 7.51 | pIC50 | 31.1 | nM | IC50 | Bioorg Med Chem (2016) 24: 2673-2680 [PMID:27131639] |
| ChEMBL | Inhibition of human recombinant EGFR T790M/L858R double mutant preincubated for 10 mins followed by FAM-labeled peptide/ATP addition measured after 1 hr by mobility shift assay | B | 7.51 | pIC50 | 31 | nM | IC50 | Eur J Med Chem (2015) 104: 115-126 [PMID:26451770] |
| ChEMBL | Inhibition of EGFR L858R/T790M double mutant phosphorylation in human NCI-H1975 cells after 2 hrs by fluorescence assay | B | 7.64 | pIC50 | 23 | nM | IC50 | J Med Chem (2013) 56: 7025-7048 [PMID:23930994] |
| ChEMBL | Inhibition of recombinant human GST-tagged wild-type EGFR cytoplasmic domain (668 to 1210 residues) expressed in baculovirus expression system using tyrosine04 peptide as substrate after 60 mins in presence of ATP by Z-LYTE assay | B | 7.74 | pIC50 | 18 | nM | IC50 | Bioorg Med Chem Lett (2018) 28: 1257-1261 [PMID:29534926] |
| ChEMBL | Inhibition of wild type N-terminal His6-tagged human EGFR kinase domain (702 to 1016 residues) expressed in Sf9 cells pre-incubated for 30 mins before ATP and substrate addition by homogeneous time-resolved FRET assay | B | 7.8 | pIC50 | 16 | nM | IC50 | Bioorg Med Chem (2015) 23: 2767-2780 [PMID:25975640] |
| ChEMBL | Inhibition of wild type EGFR (unknown origin) expressed in Sf9 cells pre-incubated for 30 mins before substrate and ATP addition by homogeneous time-resolved FRET assay | B | 8 | pIC50 | 10 | nM | IC50 | J Med Chem (2015) 58: 6844-6863 [PMID:26275028] |
| ChEMBL | Inhibition of EGFR T790M/L858R mutant (unknown origin) expressed in baculovirus expression system after 1 hr by ELISA | B | 8.01 | pIC50 | 9.7 | nM | IC50 | Bioorg Med Chem (2016) 24: 2673-2680 [PMID:27131639] |
| ChEMBL | EGFR kinase assay: In vitro enzyme activity assay: wild-type and various mutants (T790M, L858R, L861Q, L858 R/T790M) EGFR, Z′-Lyte Kinase Assay Kit were purchased from Invitrogen. 10 concentration gradients, from 5.1×10−11 mol/L to 1.0×10−6 mol/L, were set for all of the compounds to be tested. Concentrations of different kinases were determined based on the optimization of experiment, and the corresponding concentrations were: EGFR (PV3872, Invitrogen) 0.287 μg/μL, EGFR-T790M (PV4803, Invitrogen) 0.174 μg/μL, EGFR-L858R (PV4128, Invitrogen) 0.054 μg/μL, EGFR-L858R/T790M (PV4879, Invitrogen) 0.055 μg/μL. Compounds were diluted for 3 times in DMSO from 5.1×10−9 M to 1×10−4 M. 4 μL of compound was dissolved in 96 μL of water, to give a 4× compound solution. 40 μM ATP was dissolved in 1.33× kinase buffer, and a kinase/peptide mixture comprising 2× kinase, 4 μM tyrosine and four peptides was prepared for use. 10 μL of kinase reaction system comprised 2.5 μL of compound solution, 5 μL of Kinase/peptide mixture, and 2.5 μL of ATP solution. 5 μL of phosphopeptide solution was used in place of kinase/peptide mixture as 100% phosphorylation control. 2.5 μL of 1.33× kinase buffer was used to replace ATP solution as 100% inhibition control, and 2.5 μL of 4% DMSO solution was used to replace compound solution as 0% inhibition control. After thoroughly mixing the solution within the plate, the plate was incubated at room temperature for 1.5 hours. 5 μL of DevelopmentSolution was added into each well, and then the plate was incubated at room temperature for another 1 hour, and non-phosphorylated peptide was cleaved within this period. Finally, the reaction was quenched by adding 5 μL of Stop Reagent. The Plate was measured with EnVision Multilabel Reader (Perkin Elmer). Experimental data were calculated by using GraphPad Prism version 4.0. | B | 8.02 | pIC50 | 9.58 | nM | IC50 | US-9670213-B2. Pteridine ketone derivative and applications thereof as EGFR, BLK, and FLT3 inhibitor (2017) |
| ChEMBL | Inhibition of EGFR kinase L858R mutant (unknown origin) after 1.5 hrs by FRET-based Z'-Lyte assay | B | 8.09 | pIC50 | 8.13 | nM | IC50 | Medchemcomm (2012) 3: 1155-1159 |
| ChEMBL | Inhibition of wild type EGFR kinase (unknown origin) after 1.5 hrs by FRET-based Z'-Lyte assay | B | 8.19 | pIC50 | 6.47 | nM | IC50 | Medchemcomm (2012) 3: 1155-1159 |
| ChEMBL | Inhibition of EGFR L858R/T790M mutant (unknown origin) using Poly(Glu,Tyr)4:1 as substrate incubated for 1 hr by ELISA | B | 8.19 | pIC50 | 6.4 | nM | IC50 | Medchemcomm (2015) 6: 1693-1697 |
| ChEMBL | Inhibition of EGFR after 1.5 hr by FRET assay | B | 8.21 | pIC50 | 6.18 | nM | IC50 | J Med Chem (2012) 55: 2711-2723 [PMID:22339342] |
| ChEMBL | Inhibition of EGFR L861Q mutant after 1.5 hr by FRET assay | B | 8.21 | pIC50 | 6.13 | nM | IC50 | J Med Chem (2012) 55: 2711-2723 [PMID:22339342] |
| ChEMBL | Inhibition of EGFR L858R mutant (unknown origin) using Tyr 4 peptide as substrate after 1.5 hrs by FRET based Z'-lyte assay | B | 8.27 | pIC50 | 5.4 | nM | IC50 | Eur J Med Chem (2013) 66: 82-90 [PMID:23792318] |
| ChEMBL | Inhibition of EGFR L858R mutant (unknown origin) after 1.5 hrs by FRET-based Z'Lyte assay | B | 8.27 | pIC50 | 5.4 | nM | IC50 | J Med Chem (2013) 56: 4738-4748 [PMID:23668441] |
| ChEMBL | Inhibition of EGFR L858R mutant after 1.5 hr by FRET assay | B | 8.27 | pIC50 | 5.37 | nM | IC50 | J Med Chem (2012) 55: 2711-2723 [PMID:22339342] |
| ChEMBL | Inhibition of GST-tagged human EGFR T790M mutant cytoplasmic domain (668 to 1210 residues) expressed in baculovirus expression system using Poly G:T (4:1) as substrate after 30 mins by Z-LYTE assay | B | 8.3 | pIC50 | 5 | nM | IC50 | Eur J Med Chem (2017) 136: 497-510 [PMID:28528303] |
| ChEMBL | Inhibition of EGFR L858R mutant (unknown origin) after 1.5 hr by FRET-based Z-lyte assay | B | 8.32 | pIC50 | 4.8 | nM | IC50 | Eur J Med Chem (2014) 77: 75-83 [PMID:24607591] |
| ChEMBL | Inhibition of N-terminal GST-tagged human recombinant EGFR L858R mutant (668 to 1210 residues) expressed in baculovirus expression system by Z'-LYTE kinase assay | B | 8.33 | pIC50 | 4.64 | nM | IC50 | Eur J Med Chem (2021) 211: 113022-113022 [PMID:33239261] |
| ChEMBL | Inhibition of N-terminal GST-tagged human recombinant EGFR cytoplasmic domain (668 to 1210 residues) expressed in baculovirus expression system by Z'-LYTE kinase assay | B | 8.34 | pIC50 | 4.59 | nM | IC50 | Eur J Med Chem (2021) 211: 113022-113022 [PMID:33239261] |
| ChEMBL | Inhibition of EGFR (unknown origin) after 1.5 hr by FRET-based Z-lyte assay | B | 8.36 | pIC50 | 4.4 | nM | IC50 | Eur J Med Chem (2014) 77: 75-83 [PMID:24607591] |
| ChEMBL | Inhibition of EGFR kinase L861Q mutant (unknown origin) after 1.5 hrs by FRET-based Z'-Lyte assay | B | 8.37 | pIC50 | 4.25 | nM | IC50 | Medchemcomm (2012) 3: 1155-1159 |
| ChEMBL | Inhibition of GST-tagged human EGFR T790M/L858R double mutant cytoplasmic domain (668 to 1210 residues) expressed in baculovirus expression system using Poly G:T (4:1) as substrate after 30 mins by Z-LYTE assay | B | 8.4 | pIC50 | 4 | nM | IC50 | Eur J Med Chem (2017) 136: 497-510 [PMID:28528303] |
| ChEMBL | Inhibition of EGFR L858R mutant (unknown origin) using Tyr 4 peptide as substrate after 1.5 hrs by FRET-based Z'-LYTE assay | B | 8.42 | pIC50 | 3.82 | nM | IC50 | J Med Chem (2013) 56: 7821-7837 [PMID:24053674] |
| ChEMBL | Inhibition of EGFR T790M mutant after 1.5 hr by FRET assay | B | 8.42 | pIC50 | 3.77 | nM | IC50 | J Med Chem (2012) 55: 2711-2723 [PMID:22339342] |
| ChEMBL | Inhibition of wild type EGFR (unknown origin) after 1.5 hrs by FRET-based Z'Lyte assay | B | 8.49 | pIC50 | 3.2 | nM | IC50 | J Med Chem (2013) 56: 4738-4748 [PMID:23668441] |
| ChEMBL | Inhibition of wild type EGFR (unknown origin) using Tyr 4 peptide as substrate after 1.5 hrs by FRET based Z'-lyte assay | B | 8.49 | pIC50 | 3.2 | nM | IC50 | Eur J Med Chem (2013) 66: 82-90 [PMID:23792318] |
| ChEMBL | Inhibition of recombinant human GST-tagged EGFR L858R/T790M mutant cytoplasmic domain (668 to 1210 residues) expressed in baculovirus expression system using tyrosine04 peptide as substrate after 60 mins in presence of ATP by Z-LYTE assay | B | 8.52 | pIC50 | 3 | nM | IC50 | Bioorg Med Chem Lett (2018) 28: 1257-1261 [PMID:29534926] |
| ChEMBL | Inhibition of N-terminal GST-tagged human recombinant EGFR L861Q mutant (668 to 1210 residues) expressed in baculovirus expression system by Z'-LYTE kinase assay | B | 8.54 | pIC50 | 2.86 | nM | IC50 | Eur J Med Chem (2021) 211: 113022-113022 [PMID:33239261] |
| ChEMBL | EGFR kinase assay: In vitro enzyme activity assay: wild-type and various mutants (T790M, L858R, L861Q, L858 R/T790M) EGFR, Z′-Lyte Kinase Assay Kit were purchased from Invitrogen. 10 concentration gradients, from 5.1×10−11 mol/L to 1.0×10−6 mol/L, were set for all of the compounds to be tested. Concentrations of different kinases were determined based on the optimization of experiment, and the corresponding concentrations were: EGFR (PV3872, Invitrogen) 0.287 μg/μL, EGFR-T790M (PV4803, Invitrogen) 0.174 μg/μL, EGFR-L858R (PV4128, Invitrogen) 0.054 μg/μL, EGFR-L858R/T790M (PV4879, Invitrogen) 0.055 μg/μL. Compounds were diluted for 3 times in DMSO from 5.1×10−9 M to 1×10−4 M. 4 μL of compound was dissolved in 96 μL of water, to give a 4× compound solution. 40 μM ATP was dissolved in 1.33× kinase buffer, and a kinase/peptide mixture comprising 2× kinase, 4 μM tyrosine and four peptides was prepared for use. 10 μL of kinase reaction system comprised 2.5 μL of compound solution, 5 μL of Kinase/peptide mixture, and 2.5 μL of ATP solution. 5 μL of phosphopeptide solution was used in place of kinase/peptide mixture as 100% phosphorylation control. 2.5 μL of 1.33× kinase buffer was used to replace ATP solution as 100% inhibition control, and 2.5 μL of 4% DMSO solution was used to replace compound solution as 0% inhibition control. After thoroughly mixing the solution within the plate, the plate was incubated at room temperature for 1.5 hours. 5 μL of DevelopmentSolution was added into each well, and then the plate was incubated at room temperature for another 1 hour, and non-phosphorylated peptide was cleaved within this period. Finally, the reaction was quenched by adding 5 μL of Stop Reagent. The Plate was measured with EnVision Multilabel Reader (Perkin Elmer). Experimental data were calculated by using GraphPad Prism version 4.0. | B | 8.59 | pIC50 | 2.6 | nM | IC50 | US-9670213-B2. Pteridine ketone derivative and applications thereof as EGFR, BLK, and FLT3 inhibitor (2017) |
| ChEMBL | Inhibition of EGFR kinase T790M mutant (unknown origin) after 1.5 hrs by FRET-based Z'-Lyte assay | B | 8.61 | pIC50 | 2.47 | nM | IC50 | Medchemcomm (2012) 3: 1155-1159 |
| ChEMBL | Inhibition of wild type EGFR (unknown origin) using Tyr 4 peptide as substrate after 1.5 hrs by FRET-based Z'-LYTE assay | B | 8.67 | pIC50 | 2.16 | nM | IC50 | J Med Chem (2013) 56: 7821-7837 [PMID:24053674] |
| ChEMBL | Inhibition of N-terminal GST-tagged human recombinant EGFR T790M mutant (668 to 1210 residues) expressed in baculovirus expression system by Z'-LYTE kinase assay | B | 8.7 | pIC50 | 1.98 | nM | IC50 | Eur J Med Chem (2021) 211: 113022-113022 [PMID:33239261] |
| ChEMBL | Inhibition of EGFR L858R/T790M mutant after 1.5 hr by FRET assay | B | 8.73 | pIC50 | 1.88 | nM | IC50 | J Med Chem (2012) 55: 2711-2723 [PMID:22339342] |
| ChEMBL | Inhibition of EGFR kinase L858R/T790M double mutant (unknown origin) after 1.5 hrs by FRET-based Z'-Lyte assay | B | 8.74 | pIC50 | 1.82 | nM | IC50 | Medchemcomm (2012) 3: 1155-1159 |
| ChEMBL | Inhibition of EGFR (unknown origin) using biotin-labelled peptide as substrate preincubated for 2 hrs followed by substrate addition and measured after 30 mins by HTRF FRET assay | B | 8.77 | pIC50 | 1.7 | nM | IC50 | J Med Chem (2019) 62: 5901-5919 [PMID:31145622] |
| ChEMBL | Inhibition of EGFR L858R mutant (unknown origin) | B | 8.96 | pIC50 | 1.1 | nM | IC50 | J Med Chem (2017) 60: 4636-4656 [PMID:28482151] |
| ChEMBL | EGFR kinase assay: In vitro enzyme activity assay: wild-type and various mutants (T790M, L858R, L861Q, L858 R/T790M) EGFR, Z′-Lyte Kinase Assay Kit were purchased from Invitrogen. 10 concentration gradients, from 5.1×10−11 mol/L to 1.0×10−6 mol/L, were set for all of the compounds to be tested. Concentrations of different kinases were determined based on the optimization of experiment, and the corresponding concentrations were: EGFR (PV3872, Invitrogen) 0.287 μg/μL, EGFR-T790M (PV4803, Invitrogen) 0.174 μg/μL, EGFR-L858R (PV4128, Invitrogen) 0.054 μg/μL, EGFR-L858R/T790M (PV4879, Invitrogen) 0.055 μg/μL. Compounds were diluted for 3 times in DMSO from 5.1×10−9 M to 1×10−4 M. 4 μL of compound was dissolved in 96 μL of water, to give a 4× compound solution. 40 μM ATP was dissolved in 1.33× kinase buffer, and a kinase/peptide mixture comprising 2× kinase, 4 μM tyrosine and four peptides was prepared for use. 10 μL of kinase reaction system comprised 2.5 μL of compound solution, 5 μL of Kinase/peptide mixture, and 2.5 μL of ATP solution. 5 μL of phosphopeptide solution was used in place of kinase/peptide mixture as 100% phosphorylation control. 2.5 μL of 1.33× kinase buffer was used to replace ATP solution as 100% inhibition control, and 2.5 μL of 4% DMSO solution was used to replace compound solution as 0% inhibition control. After thoroughly mixing the solution within the plate, the plate was incubated at room temperature for 1.5 hours. 5 μL of DevelopmentSolution was added into each well, and then the plate was incubated at room temperature for another 1 hour, and non-phosphorylated peptide was cleaved within this period. Finally, the reaction was quenched by adding 5 μL of Stop Reagent. The Plate was measured with EnVision Multilabel Reader (Perkin Elmer). Experimental data were calculated by using GraphPad Prism version 4.0. | B | 8.99 | pIC50 | 1.02 | nM | IC50 | US-9670213-B2. Pteridine ketone derivative and applications thereof as EGFR, BLK, and FLT3 inhibitor (2017) |
| ChEMBL | Inhibition of EGFR L858R/T790M mutant (unknown origin) expressed in Sf9 cells pre-incubated for 60 mins before substrate and ATP addition by homogeneous time-resolved FRET assay | B | 9 | pIC50 | <1 | nM | IC50 | J Med Chem (2015) 58: 6844-6863 [PMID:26275028] |
| ChEMBL | Inhibition of recombinant human GST-tagged EGFR L858R/T790M double mutant expressed in baculovirus expression system preincubated for 30 mins followed by ATP and TK-substrate addition measured after 20 mins by HTRF assay | B | 9 | pIC50 | <1 | nM | IC50 | J Med Chem (2017) 60: 7725-7744 [PMID:28853575] |
| ChEMBL | Inhibition of human recombinant GST-tagged EGFR L858R mutant expressed in baculovirus expression system preincubated for 30 mins followed by ATP and TK-substrate addition measured after 15 mins by HTRF assay | B | 9 | pIC50 | <1 | nM | IC50 | J Med Chem (2017) 60: 7725-7744 [PMID:28853575] |
| ChEMBL | Inhibition of human N-terminal GST-fused EGFR T790M/L858R double mutant (669 to 1210 residues) expressed in baculovirus using TK-substrate-biotin preincubated for 30 mins followed by substrate addition measured after 20 mins by HTFR assay | B | 9 | pIC50 | <1 | nM | IC50 | J Med Chem (2017) 60: 5613-5637 [PMID:28603991] |
| ChEMBL | Inhibition of human N-terminal GST-fused EGFR L858R mutant (669 to 1210 residues) expressed in baculovirus using TK-substrate-biotin preincubated for 30 mins followed by substrate addition measured after 15 mins by HTFR assay | B | 9 | pIC50 | <1 | nM | IC50 | J Med Chem (2017) 60: 5613-5637 [PMID:28603991] |
| ChEMBL | Inhibition of EGFR L858R mutant (unknown origin) expressed in Sf9 cells pre-incubated for 60 mins before substrate and ATP addition by homogeneous time-resolved FRET assay | B | 9 | pIC50 | <1 | nM | IC50 | J Med Chem (2015) 58: 6844-6863 [PMID:26275028] |
| ChEMBL | Inhibition of recombinant human N-terminal GST tagged EGFR L858R mutant (669 to 1210 residues) expressed in insect expression system using peptide as substrate incubated for 2 hrs followed by substrate addition and measured after 30 mins by TR-FRET assay | B | 9.1 | pIC50 | 0.8 | nM | IC50 | Bioorg Med Chem (2018) 26: 1740-1750 [PMID:29523467] |
| ChEMBL | Inhibition of EGFR L858R/T790M mutant (unknown origin) using Tyr 4 peptide as substrate after 1.5 hrs by FRET based Z'-lyte assay | B | 9.1 | pIC50 | 0.8 | nM | IC50 | Eur J Med Chem (2013) 66: 82-90 [PMID:23792318] |
| ChEMBL | Inhibition of EGFR L858R/T790M double mutant (unknown origin) after 1.5 hrs by FRET-based Z'Lyte assay | B | 9.1 | pIC50 | 0.8 | nM | IC50 | J Med Chem (2013) 56: 4738-4748 [PMID:23668441] |
| ChEMBL | Inhibition of EGFR L858R/T790M mutant (unknown origin) after 1.5 hr by FRET-based Z-lyte assay | B | 9.15 | pIC50 | 0.7 | nM | IC50 | Eur J Med Chem (2014) 77: 75-83 [PMID:24607591] |
| ChEMBL | Inhibition of EGFR T790M/L858R double mutant (unknown origin) using Tyr 4 peptide as substrate after 1.5 hrs by FRET-based Z'-LYTE assay | B | 9.18 | pIC50 | 0.66 | nM | IC50 | J Med Chem (2013) 56: 7821-7837 [PMID:24053674] |
| ChEMBL | Inhibition of EGFR L858R/T790M double mutant (unknown origin) | B | 9.3 | pIC50 | <0.5 | nM | IC50 | J Med Chem (2017) 60: 4636-4656 [PMID:28482151] |
| ChEMBL | Inhibition of GST-tagged human recombinant EGFR catalytic domain (668 to 1210 amino acids) L858R mutant expressed in Sf9 cells pre-incubated for 30 mins before ATP and substrate addition by homogeneous time-resolved FRET assay | B | 9.38 | pIC50 | 0.42 | nM | IC50 | Bioorg Med Chem (2015) 23: 2767-2780 [PMID:25975640] |
| ChEMBL | Inhibition of recombinant human N-terminal GST tagged EGFR d746-750 mutant (669 to 1210 residues) expressed in insect expression system using peptide as substrate incubated for 2 hrs followed by substrate addition and measured after 30 mins by TR-FRET assay | B | 9.52 | pIC50 | 0.3 | nM | IC50 | Bioorg Med Chem (2018) 26: 1740-1750 [PMID:29523467] |
| ChEMBL | Inhibition of GST-tagged human recombinant EGFR catalytic domain (668 to 1210 amino acids) T790M/L858R mutant expressed in Sf9 cells pre-incubated for 30 mins before ATP and substrate addition by homogeneous time-resolved FRET assay | B | 9.72 | pIC50 | 0.19 | nM | IC50 | Bioorg Med Chem (2015) 23: 2767-2780 [PMID:25975640] |
| ChEMBL | Inhibition of recombinant human N-terminal GST tagged EGFR L858R/T790M double mutant (669 to 1210 residues) expressed in insect expression system using peptide as substrate incubated for 2 hrs followed by substrate addition and measured after 30 mins by TR-FRET assay | B | 10.4 | pIC50 | 0.04 | nM | IC50 | Bioorg Med Chem (2018) 26: 1740-1750 [PMID:29523467] |
| ChEMBL | Inhibition of wild type EGFR (unknown origin) transfected in mouse BAF3 cells assessed as reduction in cell viability after 72 hrs by MTS assay | B | 6 | pEC50 | 989 | nM | EC50 | Eur J Med Chem (2017) 136: 497-510 [PMID:28528303] |
| ChEMBL | Inhibition of EGFR exon19 deletion mutant (unknown origin) transfected in mouse BAF3 cells assessed as reduction in cell viability after 72 hrs by MTS assay | B | 7.17 | pEC50 | 68 | nM | EC50 | Eur J Med Chem (2017) 136: 497-510 [PMID:28528303] |
| ChEMBL | Inhibition of EGFR T790M/L858R double mutant (unknown origin) transfected in mouse BAF3 cells assessed as reduction in cell viability after 72 hrs by MTS assay | B | 7.51 | pEC50 | 31 | nM | EC50 | Eur J Med Chem (2017) 136: 497-510 [PMID:28528303] |
| ChEMBL | Inhibition of EGFR T790M exon19 deletion double mutant (unknown origin) transfected in mouse BAF3 cells assessed as reduction in cell viability after 72 hrs by MTS assay | B | 7.57 | pEC50 | 27 | nM | EC50 | Eur J Med Chem (2017) 136: 497-510 [PMID:28528303] |
| ChEMBL | Inhibition of EGFR L858R mutant (unknown origin) transfected in mouse BAF3 cells assessed as reduction in cell viability after 72 hrs by MTS assay | B | 7.59 | pEC50 | 26 | nM | EC50 | Eur J Med Chem (2017) 136: 497-510 [PMID:28528303] |
| protein tyrosine kinase 2/Focal adhesion kinase 1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2695] [GtoPdb: 2180] [UniProtKB: Q05397] | ||||||||
| ChEMBL | Binding affinity to FAK (unknown origin) assessed as dissociation constant | B | 7.11 | pKd | 78 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| NUAK family kinase 1/NUAK family SNF1-like kinase 1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL5784] [GtoPdb: 2129] [UniProtKB: O60285] | ||||||||
| GtoPdb | - | - | 7.04 | pKd | 91 | nM | Kd | Nature (2009) 462: 1070-4 [PMID:20033049] |
| ChEMBL | Binding affinity to ARK5 (unknown origin) assessed as dissociation constant | B | 7.04 | pKd | 91 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| ChEMBL | Inhibition of N-terminal GST-tagged wild type human NUAK1 expressed in Escherichia coli DH5alpha incubated for 30 mins in presence of gamma-[32P]ATP by cerenkov counting analysis | B | 7.6 | pIC50 | 25 | nM | IC50 | J Med Chem (2021) 64: 2-25 [PMID:33356242] |
| NUAK family, SNF1-like kinase, 2/NUAK family SNF1-like kinase 2 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL5698] [GtoPdb: 2130] [UniProtKB: Q9H093] | ||||||||
| ChEMBL | Binding affinity to SNARK (unknown origin) assessed as dissociation constant | B | 8 | pKd | 10 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| fms related receptor tyrosine kinase 3/Receptor-type tyrosine-protein kinase FLT3 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL1974] [GtoPdb: 1807] [UniProtKB: P36888] | ||||||||
| ChEMBL | Binding affinity to FLT3 D835Y mutant (unknown origin) assessed as dissociation constant | B | 7.48 | pKd | 33 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| erb-b2 receptor tyrosine kinase 4/Receptor tyrosine-protein kinase erbB-4 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL3009] [GtoPdb: 1799] [UniProtKB: Q15303] | ||||||||
| GtoPdb | - | - | 7.55 | pKd | 28 | nM | Kd | Nature (2009) 462: 1070-4 [PMID:20033049] |
| ChEMBL | Binding affinity to ERBB4 (unknown origin) assessed as dissociation constant | B | 7.55 | pKd | 28 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| doublecortin like kinase 3/Serine/threonine-protein kinase DCLK3 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL6123] [GtoPdb: 2007] [UniProtKB: Q9C098] | ||||||||
| ChEMBL | Binding affinity to DCAMKL3 (unknown origin) assessed as dissociation constant | B | 6.04 | pKd | 910 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| polo like kinase 4/Serine/threonine-protein kinase PLK4 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL3788] [GtoPdb: 2171] [UniProtKB: O00444] | ||||||||
| ChEMBL | Binding affinity to PLK4 (unknown origin) assessed as dissociation constant | B | 7 | pKd | 100 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| ABL proto-oncogene 1, non-receptor tyrosine kinase/Tyrosine-protein kinase ABL1 in Mouse (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL3099] [GtoPdb: 1923] [UniProtKB: P00520] | ||||||||
| ChEMBL | Inhibition of ABL T315I mutant in mouse BAF3 cells assessed as reduction in cell viability after 72 hrs by MTT assay | B | 4.79 | pIC50 | 16280 | nM | IC50 | Bioorg Med Chem Lett (2017) 27: 4832-4837 [PMID:28974338] |
| ChEMBL | Inhibition of ABL in mouse BAF3 cells assessed as reduction in cell viability after 72 hrs by MTT assay | B | 5.01 | pIC50 | 9690 | nM | IC50 | Bioorg Med Chem Lett (2017) 27: 4832-4837 [PMID:28974338] |
| BLK proto-oncogene, Src family tyrosine kinase/Tyrosine-protein kinase Blk in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2250] [GtoPdb: 1940] [UniProtKB: P51451] | ||||||||
| ChEMBL | Binding affinity to BLK (unknown origin) assessed as dissociation constant | B | 6.66 | pKd | 220 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| GtoPdb | - | - | 6.7 | pKd | 200 | nM | Kd | Nature (2009) 462: 1070-4 [PMID:20033049] |
| ChEMBL | Inhibition of TEL fused BLK (unknown origin) transformed in mouse BaF3 cells | B | 6.12 | pIC50 | 756 | nM | IC50 | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| Bruton tyrosine kinase/Tyrosine-protein kinase BTK in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL5251] [GtoPdb: 1948] [UniProtKB: Q06187] | ||||||||
| GtoPdb | - | - | 7.37 | pKd | 43 | nM | Kd | Nature (2009) 462: 1070-4 [PMID:20033049] |
| ChEMBL | Binding affinity to BTK (unknown origin) assessed as dissociation constant | B | 7.44 | pKd | 36 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| IL2 inducible T cell kinase/Tyrosine-protein kinase ITK/TSK in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2959] [GtoPdb: 2046] [UniProtKB: Q08881] | ||||||||
| ChEMBL | Binding affinity to ITK (unknown origin) assessed as dissociation constant | B | 7.37 | pKd | 43 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| GtoPdb | - | - | 7.37 | pKd | 43 | nM | Kd | Nature (2009) 462: 1070-4 [PMID:20033049] |
| Janus kinase 2/Tyrosine-protein kinase JAK2 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2971] [GtoPdb: 2048] [UniProtKB: O60674] | ||||||||
| ChEMBL | Inhibition of TEL fused JAK2 (unknown origin) transformed in mouse BaF3 cells | B | 5.42 | pIC50 | 3760 | nM | IC50 | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| Janus kinase 3/Tyrosine-protein kinase JAK3 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2148] [GtoPdb: 2049] [UniProtKB: P52333] | ||||||||
| ChEMBL | Binding affinity to JAK3 (unknown origin) | B | 6.82 | pKd | 150 | nM | Kd | J Med Chem (2015) 58: 6589-6606 [PMID:26258521] |
| ChEMBL | Binding affinity to JAK3 (unknown origin) assessed as dissociation constant | B | 6.82 | pKd | 150 | nM | Kd | Nature (2009) 462: 1070-1074 [PMID:20033049] |
| ChEMBL | Inhibition of TEL fused JAK3 (unknown origin) transformed in mouse BaF3 cells | B | 5.55 | pIC50 | 2820 | nM | IC50 | Nature (2009) 462: 1070-1074 [PMID:20033049] |
ChEMBL data shown on this page come from version 36:
Zdrazil B, Felix E, Hunter F, Manners EJ, Blackshaw J, Corbett S, de Veij M, Ioannidis H, Lopez DM, Mosquera JF, Magarinos MP, Bosc N, Arcila R, Kizilören T, Gaulton A, Bento AP, Adasme MF, Monecke P, Landrum GA, Leach AR. (2024). The ChEMBL Database in 2023: a drug discovery platform spanning multiple bioactivity data types and time periods. Nucleic Acids Res., 52(D1). DOI: 10.1093/nar/gkad1004. [EPMCID:10767899] [PMID:37933841]
Davies M, Nowotka M, Papadatos G, Dedman N, Gaulton A, Atkinson F, Bellis L, Overington JP. (2015) 'ChEMBL web services: streamlining access to drug discovery data and utilities.' Nucleic Acids Res., 43(W1). DOI: 10.1093/nar/gkv352. [EPMCID:25883136]
