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Assay Detail

CHEMBL5732901

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

Binding
LANCE ULight TR-FRET Kinase Assay: CDK9 enzyme activities were measured using LANCE ULight TR-FRET kinase assay reagents (PerkinElmer, Waltham, Mass.). Compounds were directly added in 100% DMSO to white low volume assay plates (Perkin Elmer Proxiplate 6008289) using a Labcyte Echo acoustic dispenser. Assay reagents in serine/threonine kinase assay buffer containing 20 mM HEPES, 10 mM MgCl2, 100 mM Na3VO4, and 0.0075% Triton X-100. were added for final reaction mixture concentrations of 1000 μM ATP, 100 nM U-light MBP peptide (Perkin Elmer TRF0109M) and reaction initiated with 4 nM CDK9/Cyclin T1 (Carna Biosciences 04-110). The kinase reaction was carried out for 30 minutes before addition of stopping buffer to a final of 20 mM EDTA and 0.5 nM of LANCE Ultra Europium anti-phospho-MBP antibody (PerkinElmer TRF0201M) in LANCE detection buffer (PerkinElmer CR97-100). The reaction was equilibrated for 1 hour and the signal read in the Perkin Elmer Envision in TR-FRET mode (excitation at 320 nm and emission at 615/665 nm).
4089
Total Activities
1352
Compounds Tested
3
Activity Types
0
Assay Parameters

Assay Information

Assay Type Binding
Organism Homo sapiens
Confidence 7 — Homologous single protein target
Curated By Autocuration

Target

CDK9/cyclin T1 (CHEMBL2111389)
Type PROTEIN COMPLEX
Organism Homo sapiens

Publication

Pyrrolo [2,3-B] pyridine CDK9 kinase inhibitors
(2017)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 1363 7.03 8.40
kon 1363 - -
k_off 1363 - -

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL6065695 3 8.40
CHEMBL3668457 3 8.40
CHEMBL5986595 3 8.22
CHEMBL5943573 3 8.15
CHEMBL3668461 3 8.10
CHEMBL3668473 3 8.10
CHEMBL5902372 3 8.05
CHEMBL5990315 3 8.05
CHEMBL3663778 3 8.00
CHEMBL3668361 6 8.00
CHEMBL5782477 3 8.00
CHEMBL3663759 3 7.96
CHEMBL3668476 3 7.96
CHEMBL3668418 3 7.92
CHEMBL5769589 3 7.92
CHEMBL3668375 3 7.92
CHEMBL3663764 3 7.92
CHEMBL3668378 3 7.89
CHEMBL3663792 3 7.85
CHEMBL3668370 3 7.85
CHEMBL3668385 3 7.85
CHEMBL3668401 3 7.85
CHEMBL5834238 3 7.85
CHEMBL5854326 3 7.85
CHEMBL5984950 3 7.85
CHEMBL3663746 3 7.82
CHEMBL3663774 3 7.82
CHEMBL3663796 3 7.82
CHEMBL3663797 3 7.82
CHEMBL3663802 3 7.82

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL6065695 IC50 = 4.0 nM 8.40
CHEMBL3668457 IC50 = 4.0 nM 8.40
CHEMBL5986595 IC50 = 6.0 nM 8.22
CHEMBL5943573 IC50 = 7.0 nM 8.15
CHEMBL3668461 IC50 = 8.0 nM 8.10
CHEMBL3668473 IC50 = 8.0 nM 8.10
CHEMBL5902372 IC50 = 9.0 nM 8.05
CHEMBL5990315 IC50 = 9.0 nM 8.05
CHEMBL3663778 IC50 = 10.0 nM 8.00
CHEMBL5782477 IC50 = 10.0 nM 8.00
CHEMBL3668361 IC50 = 10.0 nM 8.00
CHEMBL3668476 IC50 = 11.0 nM 7.96
CHEMBL3663759 IC50 = 11.0 nM 7.96
CHEMBL3668418 IC50 = 12.0 nM 7.92
CHEMBL3668375 IC50 = 12.0 nM 7.92
CHEMBL3663764 IC50 = 12.0 nM 7.92
CHEMBL5769589 IC50 = 12.0 nM 7.92
CHEMBL3668378 IC50 = 13.0 nM 7.89
CHEMBL3668385 IC50 = 14.0 nM 7.85
CHEMBL5854326 IC50 = 14.0 nM 7.85
CHEMBL3663792 IC50 = 14.0 nM 7.85
CHEMBL5984950 IC50 = 14.0 nM 7.85
CHEMBL5834238 IC50 = 14.0 nM 7.85
CHEMBL3668370 IC50 = 14.0 nM 7.85
CHEMBL3668401 IC50 = 14.0 nM 7.85
CHEMBL3663746 IC50 = 15.0 nM 7.82
CHEMBL3668427 IC50 = 15.0 nM 7.82
CHEMBL5947200 IC50 = 15.0 nM 7.82
CHEMBL5905192 IC50 = 15.0 nM 7.82
CHEMBL3663802 IC50 = 15.0 nM 7.82
CHEMBL3663774 IC50 = 15.0 nM 7.82
CHEMBL3663796 IC50 = 15.0 nM 7.82
CHEMBL3663797 IC50 = 15.0 nM 7.82
CHEMBL5905080 IC50 = 16.0 nM 7.80
CHEMBL5852792 IC50 = 16.0 nM 7.80
CHEMBL3663743 IC50 = 16.0 nM 7.80
CHEMBL6010037 IC50 = 16.0 nM 7.80
CHEMBL3668350 IC50 = 17.0 nM 7.77
CHEMBL3663777 IC50 = 17.0 nM 7.77
CHEMBL5949412 IC50 = 17.0 nM 7.77
CHEMBL3663769 IC50 = 17.0 nM 7.77
CHEMBL5940848 IC50 = 17.0 nM 7.77
CHEMBL6035846 IC50 = 17.0 nM 7.77
CHEMBL3663799 IC50 = 17.0 nM 7.77
CHEMBL3668421 IC50 = 17.0 nM 7.77
CHEMBL5875544 IC50 = 17.0 nM 7.77
CHEMBL6024010 IC50 = 17.0 nM 7.77
CHEMBL6065926 IC50 = 17.0 nM 7.77
CHEMBL3668480 IC50 = 17.0 nM 7.77
CHEMBL3663775 IC50 = 18.0 nM 7.75