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Assay Detail

CHEMBL3707623

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

Binding
HTRF Assay: The ability of compounds to inhibit the activity of JAK1, JAK2, JAK3, and Tyk2 was measured using a recombinant purified GST-tagged catalytic domain for each enzyme (Invitrogen JAK1 #M4290, JAK2 #M4290, JAK3 #M4290, Tyk2 #M4290) in an HTRF format biochemical assay. The reactions employed a common peptide substrate, LCB-EQEDEPEGDYFEWLW-NH2 (in-house). The basic assay protocol is as follows: First, 250 mL of diluted compounds in DMSO were dispensed into the wells of a dry 384-well Black plate (Greiner #781076) using a Labcyte Echo 555 acoustic dispenser. Subsequent reagent additions employed an Agilent Bravo. Next, 18 uL of 1.11x enzyme and 1.11x substrate in 1x assay buffer (Invitrogen kinase buffer #PV3189, 2 mM DTT, 0.05% BSA) were added to the wells and shaken and then preincubated for 30 minutes at ambient temperature to allow compound binding to equilibrate. After equilibration, 2 uL of 10xATP in 1x assay buffer was added to initiate the kinase reaction.
596
Total Activities
561
Compounds Tested
1
Activity Types
0
Assay Parameters

Assay Information

Assay Type Binding
Organism Homo sapiens
Confidence 9 — Direct single protein target
Curated By Autocuration

Target

Tyrosine-protein kinase JAK2 (CHEMBL2971)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Cycloalkylnitrile pyrazole carboxamides as janus kinase inhibitors
(2015)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 596 7.59 9.18

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL3699558 1 9.18
CHEMBL3699521 1 9.10
CHEMBL3702904 1 9.05
CHEMBL3647859 1 9.02
CHEMBL3702907 1 9.02
CHEMBL3702903 1 9.00
CHEMBL3702910 1 9.00
CHEMBL3647845 1 9.00
CHEMBL3702889 1 9.00
CHEMBL3702920 1 9.00
CHEMBL3702919 1 9.00
CHEMBL3702887 1 9.00
CHEMBL3647728 1 9.00
CHEMBL3702914 1 9.00
CHEMBL3647736 1 9.00
CHEMBL3699531 1 9.00
CHEMBL3703042 1 9.00
CHEMBL3647923 2 9.00
CHEMBL3702905 1 9.00
CHEMBL3647858 1 8.70
CHEMBL3699528 1 8.70
CHEMBL3699499 2 8.70
CHEMBL3647835 2 8.70
CHEMBL3647922 2 8.70
CHEMBL3647918 1 8.70
CHEMBL3647751 1 8.70
CHEMBL3702886 1 8.70
CHEMBL3647856 1 8.70
CHEMBL3702883 1 8.70
CHEMBL3702947 1 8.70

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL3699558 IC50 = 0.66 nM 9.18
CHEMBL3699521 IC50 = 0.79 nM 9.10
CHEMBL3702904 IC50 = 0.9 nM 9.05
CHEMBL3702907 IC50 = 0.95 nM 9.02
CHEMBL3647859 IC50 = 0.95 nM 9.02
CHEMBL3702910 IC50 = 1.0 nM 9.00
CHEMBL3702903 IC50 = 1.0 nM 9.00
CHEMBL3702889 IC50 = 1.0 nM 9.00
CHEMBL3699531 IC50 = 1.0 nM 9.00
CHEMBL3647728 IC50 = 1.0 nM 9.00
CHEMBL3702920 IC50 = 1.0 nM 9.00
CHEMBL3647923 IC50 = 1.0 nM 9.00
CHEMBL3647845 IC50 = 1.0 nM 9.00
CHEMBL3702905 IC50 = 1.0 nM 9.00
CHEMBL3702914 IC50 = 1.0 nM 9.00
CHEMBL3647736 IC50 = 1.0 nM 9.00
CHEMBL3703042 IC50 = 1.0 nM 9.00
CHEMBL3702919 IC50 = 1.0 nM 9.00
CHEMBL3702887 IC50 = 1.0 nM 9.00
CHEMBL3702953 IC50 = 2.0 nM 8.70
CHEMBL3702947 IC50 = 2.0 nM 8.70
CHEMBL3702883 IC50 = 2.0 nM 8.70
CHEMBL3702886 IC50 = 2.0 nM 8.70
CHEMBL3699551 IC50 = 2.0 nM 8.70
CHEMBL3699537 IC50 = 2.0 nM 8.70
CHEMBL3699528 IC50 = 2.0 nM 8.70
CHEMBL3647919 IC50 = 2.0 nM 8.70
CHEMBL3647751 IC50 = 2.0 nM 8.70
CHEMBL3647858 IC50 = 2.0 nM 8.70
CHEMBL3647856 IC50 = 2.0 nM 8.70
CHEMBL3699499 IC50 = 2.0 nM 8.70
CHEMBL3647861 IC50 = 2.0 nM 8.70
CHEMBL3647918 IC50 = 2.0 nM 8.70
CHEMBL3647922 IC50 = 2.0 nM 8.70
CHEMBL3702902 IC50 = 2.0 nM 8.70
CHEMBL3702908 IC50 = 2.0 nM 8.70
CHEMBL3702912 IC50 = 2.0 nM 8.70
CHEMBL3647835 IC50 = 2.0 nM 8.70
CHEMBL3703048 IC50 = 2.0 nM 8.70
CHEMBL3647727 IC50 = 2.0 nM 8.70
CHEMBL3699538 IC50 = 3.0 nM 8.52
CHEMBL3647835 IC50 = 3.0 nM 8.52
CHEMBL3699500 IC50 = 3.0 nM 8.52
CHEMBL3699530 IC50 = 3.0 nM 8.52
CHEMBL3702957 IC50 = 3.0 nM 8.52
CHEMBL3647771 IC50 = 3.0 nM 8.52
CHEMBL3647852 IC50 = 3.0 nM 8.52
CHEMBL3702951 IC50 = 3.0 nM 8.52
CHEMBL3647924 IC50 = 3.0 nM 8.52
CHEMBL3703051 IC50 = 3.0 nM 8.52