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Assay Detail

CHEMBL3706338

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Binding
Inhibition Assay: A recombinant retrovirus was created from expression plasmid FLAG-EML4-ALKv1/pMX-iresCD8 in which cDNA for EMLA-ALK fusion protein v1 was integrated, and injected into mouse lymphoid cell line BA/F3 cells. Using a magnetic bead reagent for cell separation and a purification column (anti-CD8 monoclonal antibody immobilized on magnetic beads and a MiniMACS purification column; both are products of MilteDyi Biotec Inc.), cell surface CD8-expressing cells were purified to establish EML4-ALK fusion protein v1-expressing BA/F3 cells. From the cells, EML4-ALK fusion protein v1 was purified and subjected to kinase activity evaluation. EML4-ALK fusion protein v1 was investigated for its phosphorylation activity toward a peptide substrate by using a kinase activity detection kit (HTRF KinEASE-TK; Cisbio Inc.). Test compounds were each added to a reaction solution containing the enzyme protein to give 8 final concentrations from 1000 nM to 0.3 nM, followed by addition of ATP.
41
Total Activities
39
Compounds Tested
1
Activity Types
0
Assay Parameters

Assay Information

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

Target

ALK tyrosine kinase receptor (CHEMBL4247)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Diamino heterocyclic carboxamide compound
(2015)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 41 8.82 9.77

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL3687205 1 9.77
CHEMBL3687207 1 9.68
CHEMBL3687206 1 9.66
CHEMBL3889838 1 9.64
CHEMBL3687201 1 9.62
CHEMBL3687204 1 9.59
CHEMBL3687211 1 9.59
CHEMBL3687202 1 9.59
CHEMBL3687198 1 9.48
CHEMBL3687209 1 9.47
CHEMBL3687193 1 9.43
CHEMBL3687210 1 9.31
CHEMBL3687200 1 9.19
CHEMBL3687197 1 9.11
CHEMBL3687212 1 9.03
CHEMBL3687208 1 9.03
CHEMBL3687189 1 9.00
CHEMBL3687216 1 9.00
CHEMBL3687223 1 8.85
CHEMBL3301622 GILTERITINIB 4.0 1 8.82
CHEMBL3687215 1 8.82
CHEMBL3975354 1 8.77
CHEMBL3687218 1 8.72
CHEMBL3687195 1 8.68
CHEMBL3687221 1 8.68
CHEMBL3687225 1 8.66
CHEMBL3687217 1 8.64
CHEMBL3687214 2 8.59
CHEMBL3687213 2 8.55
CHEMBL3687194 1 8.55

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL3687205 IC50 = 0.17 nM 9.77
CHEMBL3687207 IC50 = 0.21 nM 9.68
CHEMBL3687206 IC50 = 0.22 nM 9.66
CHEMBL3889838 IC50 = 0.23 nM 9.64
CHEMBL3687201 IC50 = 0.24 nM 9.62
CHEMBL3687204 IC50 = 0.26 nM 9.59
CHEMBL3687211 IC50 = 0.26 nM 9.59
CHEMBL3687202 IC50 = 0.26 nM 9.59
CHEMBL3687198 IC50 = 0.33 nM 9.48
CHEMBL3687209 IC50 = 0.34 nM 9.47
CHEMBL3687193 IC50 = 0.37 nM 9.43
CHEMBL3687210 IC50 = 0.49 nM 9.31
CHEMBL3687200 IC50 = 0.65 nM 9.19
CHEMBL3687197 IC50 = 0.77 nM 9.11
CHEMBL3687208 IC50 = 0.94 nM 9.03
CHEMBL3687212 IC50 = 0.93 nM 9.03
CHEMBL3687189 IC50 = 0.99 nM 9.00
CHEMBL3687216 IC50 = 1.0 nM 9.00
CHEMBL3687223 IC50 = 1.4 nM 8.85
CHEMBL3687215 IC50 = 1.5 nM 8.82
CHEMBL3301622 GILTERITINIB IC50 = 1.5 nM 8.82
CHEMBL3975354 IC50 = 1.7 nM 8.77
CHEMBL3687218 IC50 = 1.9 nM 8.72
CHEMBL3687221 IC50 = 2.1 nM 8.68
CHEMBL3687195 IC50 = 2.1 nM 8.68
CHEMBL3687225 IC50 = 2.2 nM 8.66
CHEMBL3687217 IC50 = 2.3 nM 8.64
CHEMBL3687214 IC50 = 2.6 nM 8.59
CHEMBL3687194 IC50 = 2.8 nM 8.55
CHEMBL3687213 IC50 = 2.8 nM 8.55
CHEMBL3687213 IC50 = 3.1 nM 8.51
CHEMBL3687214 IC50 = 4.1 nM 8.39
CHEMBL3687224 IC50 = 4.5 nM 8.35
CHEMBL3687191 IC50 = 5.3 nM 8.28
CHEMBL3687220 IC50 = 7.8 nM 8.11
CHEMBL3687190 IC50 = 8.9 nM 8.05
CHEMBL3687219 IC50 = 11.0 nM 7.96
CHEMBL3687222 IC50 = 11.0 nM 7.96
CHEMBL3687188 IC50 = 17.0 nM 7.77
CHEMBL3687199 IC50 = 17.0 nM 7.77
CHEMBL3687192 IC50 = 76.0 nM 7.12