High-throughput screening of novel antagonists on melanin-concentrating hormone receptor-1.
OBJECTIVE
To find new antagonists on human melanin-concentrating hormone receptor-1 (MCHR-1) through high-throughput screening (HTS) of a diverse compound library.


METHODS
MCHR-1, [3H]SNAP7941, and FlashBlue G-protein-coupled receptor beads were used to measure the receptor-binding activities of various compounds based on scintillation proximity assay (SPA) technology. The guanosine 5'(gamma-[35S]thio) triphosphate ([35S]GTPgammaS) binding assay was subsequently applied to functionally characterize the "hits" identified by the HTS campaign.


RESULTS
Of the 48,240 compounds screened with the SPA method, 12 hits were confirmed to possess MCHR-1 binding activities, 8 were functionally studied subsequently with the [35S]GTPgammaS binding assay, and only 1 compound (NC127816) displayed moderate human MCHR-1 binding affinity (Ki=115.7 nmol/L) and relatively potent antagonism (KB=23.8 nmol/L). This compound shares a novel scaffold (1-ethoxy-2H-2-aza-1-phospha-naphthalene 1-oxide) with 3 other analogs in the group.


CONCLUSIONS
Considering the marked difference in molecular shape and electrostatic status between NC127816 and the structures reported elsewhere, we anticipate that its derivatives may represent a new class of potent MCHR-1 modulators.
Yan
Jian-hua
JH
The National Center for Drug Screening and the State Key Laboratory of New Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
Li
Qun-yi
QY
Boutin
Jean A
JA
Renard
M Pierre
MP
Ding
Yi-xiang
YX
Hao
Xiao-jiang
XJ
Zhao
Wei-min
WM
Wang
Ming-wei
MW
D001667:Binding, Competitive; D002460:Cell Line; D004353:Drug Evaluation, Preclinical; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006801:Humans; D007202:Indicators and Reagents; D008024:Ligands; D010880:Piperidines / Q000378:metabolism; D011743:Pyrimidines / Q000378:metabolism; D017481:Receptors, Somatostatin / Q000037:antagonists & inhibitors*
D007202:Indicators and Reagents; D008024:Ligands; C103263:MCHR1 protein, human; D010880:Piperidines; D011743:Pyrimidines; D017481:Receptors, Somatostatin; C464854:SNAP7941; D016244:Guanosine 5'-O-(3-Thiotriphosphate)