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Assay Detail

CHEMBL3889179

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

Functional
Aequorin Functional Assay: Changes in intracellular calcium levels are a recognized indicator of G protein-coupled receptor activity. The efficacy of compounds of the invention to inhibit NKA-mediated NK-3 receptor activation was assessed by an in vitro Aequorin functional assay. Chinese Hamster Ovary recombinant cells expressing the human NK3 receptor and a construct that encodes the photoprotein apoaequorin were used for this assay. In the presence of the cofactor coelenterazine, apoaequorin emits a measurable luminescence that is proportional to the amount of intracellular (cytoplasmic) free calcium. Antagonist Testing: the antagonist activity of compounds of the invention is measured following pre-incubation (3 minutes) of the compound with the cells, followed by addition of the reference agonist (NKA) at a final concentration equivalent to the EC80 (3 nM) and recording of emitted light (FDSS 6000 Hamamatsu) over the subsequent 90-second period. The intensity of the emitted light is integrated using the reader software. Compound antagonist activity is measured based on the inhibition of the luminescence response to the addition of Neurokinin A.  Inhibition curves are obtained for compounds of the invention and the concentrations of compounds which inhibit 50% of reference agonist response (IC50) were determined (see results in table 5 below). The IC50 values shown in table 5 indicate that compounds of the invention are potent NK-3 antagonist compounds.
31
Total Activities
31
Compounds Tested
1
Activity Types
0
Assay Parameters

Assay Information

Assay Type Functional
Organism Homo sapiens
Confidence 8 — Homologous single protein target
Curated By Autocuration

Target

Neuromedin-K receptor (CHEMBL4429)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Chiral N-acyl-5,6,7(8-substituted)-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazines as selective NK-3 receptor antagonists, pharmaceutical composition, methods for use in NK-3 receptor mediated disorders and chiral synthesis thereof
(2016)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 31 7.83 8.70

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL3422018 1 8.70
CHEMBL3422015 1 8.70
CHEMBL4112270 1 8.52
CHEMBL4111118 1 8.52
CHEMBL4111525 1 8.52
CHEMBL4109138 1 8.15
CHEMBL4115030 1 8.15
CHEMBL3422016 1 8.10
CHEMBL4112531 1 8.05
CHEMBL4112776 1 8.00
CHEMBL3939146 1 8.00
CHEMBL4109230 1 7.96
CHEMBL3422007 1 7.96
CHEMBL4106444 1 7.92
CHEMBL3422010 1 7.80
CHEMBL4107668 1 7.75
CHEMBL4114218 1 7.75
CHEMBL3964222 1 7.75
CHEMBL3940252 1 7.70
CHEMBL4112613 1 7.68
CHEMBL4107180 1 7.55
CHEMBL4110178 1 7.52
CHEMBL3952660 1 7.48
CHEMBL3422017 1 7.48
CHEMBL4108578 1 7.47
CHEMBL4110064 1 7.43
CHEMBL3917687 1 7.33
CHEMBL3908229 1 7.30
CHEMBL4112541 1 7.29
CHEMBL4113811 1 7.24

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL3422018 IC50 = 2.0 nM 8.70
CHEMBL3422015 IC50 = 2.0 nM 8.70
CHEMBL4112270 IC50 = 3.0 nM 8.52
CHEMBL4111118 IC50 = 3.0 nM 8.52
CHEMBL4111525 IC50 = 3.0 nM 8.52
CHEMBL4109138 IC50 = 7.0 nM 8.15
CHEMBL4115030 IC50 = 7.0 nM 8.15
CHEMBL3422016 IC50 = 8.0 nM 8.10
CHEMBL4112531 IC50 = 9.0 nM 8.05
CHEMBL4112776 IC50 = 10.0 nM 8.00
CHEMBL3939146 IC50 = 10.0 nM 8.00
CHEMBL4109230 IC50 = 11.0 nM 7.96
CHEMBL3422007 IC50 = 11.0 nM 7.96
CHEMBL4106444 IC50 = 12.0 nM 7.92
CHEMBL3422010 IC50 = 16.0 nM 7.80
CHEMBL4107668 IC50 = 18.0 nM 7.75
CHEMBL4114218 IC50 = 18.0 nM 7.75
CHEMBL3964222 IC50 = 18.0 nM 7.75
CHEMBL3940252 IC50 = 20.0 nM 7.70
CHEMBL4112613 IC50 = 21.0 nM 7.68
CHEMBL4107180 IC50 = 28.0 nM 7.55
CHEMBL4110178 IC50 = 30.0 nM 7.52
CHEMBL3952660 IC50 = 33.0 nM 7.48
CHEMBL3422017 IC50 = 33.0 nM 7.48
CHEMBL4108578 IC50 = 34.0 nM 7.47
CHEMBL4110064 IC50 = 37.0 nM 7.43
CHEMBL3917687 IC50 = 47.0 nM 7.33
CHEMBL3908229 IC50 = 50.0 nM 7.30
CHEMBL4112541 IC50 = 51.0 nM 7.29
CHEMBL4113811 IC50 = 58.0 nM 7.24
CHEMBL3422012 IC50 = 83.0 nM 7.08