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Assay Detail

CHEMBL5732865

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Binding
CDK8/Cyclin C Binding Assay: The binding assay relies on the LanthaScreen™ Eu-Kinase Binding Assay (Invitrogen. This is a kinase assay platform based on measuring the binding and displacement of an Alexa Fluor® 647 conjugate of an ATP-competitive kinase inhibitor (Kinase Tracer 236, PV5592) at a kinase active site. Binding of the tracer to the kinase is detected by addition of a europium (Eu)-labelled anti-His antibody (Invitrogen PV 5596), which specifically labels the kinase of interest. This binding results in a high degree of fluorescence resonance energy transfer (FRET), whereas displacement of the tracer with a kinase inhibitor results in a loss of FRET.The enzyme has been purchase from Invitrogen (PV4402) as a dimer of full-length His-tagged recombinant human proteins.Assay conditions were as indicated by the kit manufacturers with the following adaptations:Assay buffer: 50 mM HEPES, pH 7.5, 1 mM EGTA, 0.01% Brij-35, 10 mM MgCl2Assay volume: 25 μlIncubation time and temperature: 60 min at 25° C.Cdk8-Cyclin C concentration: 5 nMTracer concentration: 10 nM(Eu)-labeled anti-His antibody concentration: 1.5 nMTested compound: Serial 1:3 dilutionsFinal DMSO concentration in the assay: 1%Assays were performed in 384-well plates. The final read out was generated using an EnVision plate reader (Perkin-Elmer). The emission ratio was calculated by dividing the acceptor/tracer emission (665 nm) by the antibody/donor emission (615 nm).
291
Total Activities
97
Compounds Tested
3
Activity Types
0
Assay Parameters

Assay Information

Assay Type Binding
Confidence 6 — Multiple protein complex / RNA
Curated By Autocuration

Target

CDK8/Cyclin C (CHEMBL3038474)
Type PROTEIN COMPLEX
Organism Homo sapiens

Publication

Condensed tricyclic compounds as protein kinase inhibitors
(2019)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 97 7.26 9.72
kon 97 - -
k_off 97 - -

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL4789125 3 9.72
CHEMBL4789603 3 9.58
CHEMBL4776812 3 9.56
CHEMBL4777356 3 9.50
CHEMBL5752178 3 9.39
CHEMBL4780792 3 9.33
CHEMBL4782696 3 8.96
CHEMBL5817322 3 8.87
CHEMBL4779550 3 8.85
CHEMBL4791276 3 8.85
CHEMBL4778840 3 8.85
CHEMBL4778755 3 8.85
CHEMBL4785713 3 8.71
CHEMBL4797707 3 8.70
CHEMBL4789863 3 8.68
CHEMBL4781993 3 8.66
CHEMBL4793447 3 8.62
CHEMBL4782944 3 8.59
CHEMBL4797007 3 8.58
CHEMBL4790936 3 8.49
CHEMBL4794862 3 8.42
CHEMBL5943185 3 8.38
CHEMBL4792305 3 8.36
CHEMBL5767742 3 8.35
CHEMBL4795220 3 8.33
CHEMBL4787064 3 8.13
CHEMBL4777501 3 8.07
CHEMBL5820678 3 8.05
CHEMBL6022963 3 8.03
CHEMBL6006771 3 7.91

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL4789125 IC50 = 0.191 nM 9.72
CHEMBL4789603 IC50 = 0.262 nM 9.58
CHEMBL4776812 IC50 = 0.278 nM 9.56
CHEMBL4777356 IC50 = 0.314 nM 9.50
CHEMBL5752178 IC50 = 0.407 nM 9.39
CHEMBL4780792 IC50 = 0.466 nM 9.33
CHEMBL4782696 IC50 = 1.11 nM 8.96
CHEMBL5817322 IC50 = 1.36 nM 8.87
CHEMBL4778755 IC50 = 1.41 nM 8.85
CHEMBL4778840 IC50 = 1.41 nM 8.85
CHEMBL4779550 IC50 = 1.41 nM 8.85
CHEMBL4791276 IC50 = 1.41 nM 8.85
CHEMBL4785713 IC50 = 1.96 nM 8.71
CHEMBL4797707 IC50 = 1.98 nM 8.70
CHEMBL4789863 IC50 = 2.1 nM 8.68
CHEMBL4781993 IC50 = 2.19 nM 8.66
CHEMBL4793447 IC50 = 2.41 nM 8.62
CHEMBL4782944 IC50 = 2.59 nM 8.59
CHEMBL4797007 IC50 = 2.65 nM 8.58
CHEMBL4790936 IC50 = 3.23 nM 8.49
CHEMBL4794862 IC50 = 3.83 nM 8.42
CHEMBL5943185 IC50 = 4.18 nM 8.38
CHEMBL4792305 IC50 = 4.33 nM 8.36
CHEMBL5767742 IC50 = 4.46 nM 8.35
CHEMBL4795220 IC50 = 4.64 nM 8.33
CHEMBL4787064 IC50 = 7.48 nM 8.13
CHEMBL4777501 IC50 = 8.5 nM 8.07
CHEMBL5820678 IC50 = 8.94 nM 8.05
CHEMBL6022963 IC50 = 9.32 nM 8.03
CHEMBL6006771 IC50 = 12.3 nM 7.91
CHEMBL6062128 IC50 = 13.3 nM 7.88
CHEMBL4789450 IC50 = 13.9 nM 7.86
CHEMBL4792156 IC50 = 14.2 nM 7.85
CHEMBL6002058 IC50 = 14.4 nM 7.84
CHEMBL4776992 IC50 = 14.4 nM 7.84
CHEMBL5831525 IC50 = 19.3 nM 7.71
CHEMBL4782056 IC50 = 20.2 nM 7.70
CHEMBL5967061 IC50 = 21.6 nM 7.67
CHEMBL6052876 IC50 = 23.9 nM 7.62
CHEMBL5967245 IC50 = 26.2 nM 7.58
CHEMBL5790753 IC50 = 29.7 nM 7.53
CHEMBL6035709 IC50 = 31.1 nM 7.51
CHEMBL6017565 IC50 = 31.9 nM 7.50
CHEMBL5755811 IC50 = 39.3 nM 7.41
CHEMBL5975092 IC50 = 52.5 nM 7.28
CHEMBL5770417 IC50 = 54.7 nM 7.26
CHEMBL5866937 IC50 = 55.5 nM 7.26
CHEMBL4789204 IC50 = 55.4 nM 7.26
CHEMBL5748809 IC50 = 61.1 nM 7.21
CHEMBL6060510 IC50 = 78.5 nM 7.11