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Assay Detail

CHEMBL5732491

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

Binding
Enzyme Inhibition Assay: The inhibitory activities of compounds of the invention against the GSK 3α enzyme isoform (Invitrogen), are evaluated by determining the level of activation/phosphorylation of the target peptide. The GSK3-α protein (500 ng/mL, 2.5 μL) is mixed with the test compound (2.5 μL at either 4 μg/mL, 0.4 μg/mL, 0.04 μg/mL, or 0.004 μg/mL) for 2 hr at RT. The FRET peptide (8 μM, 2.5 μL), which is a phosphorylation target for GSK3α, and ATP (40 μM, 2.5 μL) are then added to the enzyme/compound mixture and the resulting mixture incubated for 1 hr. Development reagent (protease, 5 μL) is added for 1 hr prior to detection in a fluorescence microplate reader (Varioskan® Flash, ThermoFisher Scientific).In all cases, the site-specific protease cleaves non-phosphorylated peptide only and eliminates the FRET signal. Phosphorylation levels of each reaction are calculated using the ratio of coumarin emission (donor) over fluorescein emission (acceptor), for which high ratios indicate high phosphorylation and low ratios indicate low phosphorylation levels.
216
Total Activities
72
Compounds Tested
3
Activity Types
0
Assay Parameters

Assay Information

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

Target

Glycogen synthase kinase-3 alpha (CHEMBL2850)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Kinase inhibitors
(2017)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 72 5.72 6.93
kon 72 - -
k_off 72 - -

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL5912287 3 6.93
CHEMBL5866923 3 6.90
CHEMBL5992634 3 6.76
CHEMBL5753503 3 6.59
CHEMBL6046794 3 6.51
CHEMBL6016170 3 6.38
CHEMBL5990752 3 6.37
CHEMBL5976851 3 6.30
CHEMBL6023399 3 6.28
CHEMBL5954666 3 6.22
CHEMBL5827694 3 6.09
CHEMBL5979374 3 6.08
CHEMBL5858089 3 6.01
CHEMBL5991930 3 5.99
CHEMBL5935125 3 5.97
CHEMBL5802500 3 5.91
CHEMBL5820101 3 5.88
CHEMBL5855478 3 5.85
CHEMBL6030562 3 5.83
CHEMBL6040369 3 5.73
CHEMBL5752694 3 5.71
CHEMBL5919556 3 5.71
CHEMBL5841040 3 5.69
CHEMBL5933911 3 5.60
CHEMBL5968094 3 5.57
CHEMBL5753333 3 5.55
CHEMBL5823917 3 5.55
CHEMBL5872801 3 5.54
CHEMBL5873953 3 5.52
CHEMBL5928253 3 5.47

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL5912287 IC50 = 117.0 nM 6.93
CHEMBL5866923 IC50 = 125.0 nM 6.90
CHEMBL5992634 IC50 = 172.0 nM 6.76
CHEMBL5753503 IC50 = 258.0 nM 6.59
CHEMBL6046794 IC50 = 306.0 nM 6.51
CHEMBL6016170 IC50 = 416.0 nM 6.38
CHEMBL5990752 IC50 = 426.0 nM 6.37
CHEMBL5976851 IC50 = 501.0 nM 6.30
CHEMBL6023399 IC50 = 531.0 nM 6.28
CHEMBL5954666 IC50 = 596.0 nM 6.22
CHEMBL5827694 IC50 = 804.0 nM 6.09
CHEMBL5979374 IC50 = 824.0 nM 6.08
CHEMBL5858089 IC50 = 973.0 nM 6.01
CHEMBL5991930 IC50 = 1026.0 nM 5.99
CHEMBL5935125 IC50 = 1071.0 nM 5.97
CHEMBL5802500 IC50 = 1230.0 nM 5.91
CHEMBL5820101 IC50 = 1327.0 nM 5.88
CHEMBL5855478 IC50 = 1399.0 nM 5.85
CHEMBL6030562 IC50 = 1489.0 nM 5.83
CHEMBL6040369 IC50 = 1850.0 nM 5.73
CHEMBL5752694 IC50 = 1931.0 nM 5.71
CHEMBL5919556 IC50 = 1935.0 nM 5.71
CHEMBL5841040 IC50 = 2053.0 nM 5.69
CHEMBL5933911 IC50 = 2536.0 nM 5.60
CHEMBL5968094 IC50 = 2666.0 nM 5.57
CHEMBL5823917 IC50 = 2806.0 nM 5.55
CHEMBL5753333 IC50 = 2824.0 nM 5.55
CHEMBL5872801 IC50 = 2903.0 nM 5.54
CHEMBL5873953 IC50 = 3027.0 nM 5.52
CHEMBL6011006 IC50 = 3352.0 nM 5.47
CHEMBL5928253 IC50 = 3387.0 nM 5.47
CHEMBL5888113 IC50 = 3724.0 nM 5.43
CHEMBL5769990 IC50 = 3796.0 nM 5.42
CHEMBL5943347 IC50 = 3842.0 nM 5.42
CHEMBL5864913 IC50 = 4611.0 nM 5.34
CHEMBL5822857 IC50 = 4688.0 nM 5.33
CHEMBL5810491 IC50 = 4826.0 nM 5.32
CHEMBL5750422 IC50 = 4817.0 nM 5.32
CHEMBL6010123 IC50 = 5257.0 nM 5.28
CHEMBL5990759 IC50 = 5445.0 nM 5.26
CHEMBL5840394 IC50 = 6360.0 nM 5.20
CHEMBL6009171 IC50 = 6829.0 nM 5.17
CHEMBL5963080 IC50 = 7232.0 nM 5.14
CHEMBL5981366 IC50 = 7258.0 nM 5.14
CHEMBL5821652 IC50 = 7365.0 nM 5.13
CHEMBL5835423 IC50 = 7740.0 nM 5.11
CHEMBL5967674 IC50 = 8173.0 nM 5.09
CHEMBL6040301 IC50 = 8651.0 nM 5.06
CHEMBL5846222 IC50 = 9544.0 nM 5.02
CHEMBL6041255 IC50 > 10000.0 nM -