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The non‐competitive inhibitory activities of morphinan and morphine derivatives at the α3β4 neuronal acetylcholine receptor
Author(s) -
Jozwiak Krzysztof,
Moaddel Ruin,
Yamaguchi Rika,
Maciuk Alexandre,
Wainer Irving W
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a245
Subject(s) - chemistry , naltrindole , nicotinic acetylcholine receptor , acetylcholine receptor , chromatography , morphine , receptor , opioid receptor , agonist , stereochemistry , pharmacology , biochemistry , medicine
Purpose 20 compounds (morphinan and morphine derivatives) were investigated as non‐competitive inhibitors of the α3β4 subtype of the neuronal nicotinic acetylcholine receptor (α3β4 nAChR) using newly developed high‐throughput method. Methods The identifications of inhibitors were performed using chromatographic experiments on an immobilizedα3β4 nAChR column combined with chemometric and molecular modeling techniques. The pharmacological effect of these agents was also determined using a functional assay (nicotine induced 86 Rb + efflux) from KXα3β4R2 cell line, the same cell line used to create the nAChR column. Results The chromatographic retention on the immobilized α3β4 column indicated that 11 of the 13 morphine derivatives had little interaction with the immobilized receptor, which was consistent with their observed IC 50 values of >200 μM. All of the morphinans and 2 of the morphine derivatives (buprenorphine and naltrindole) were significantly retained on the chromatographic column and had IC 50 values < 60 μM. Molecular modeling techniques were used to determine that the morphinans bound to the α3β4 nAChR at a hydrophobic pocket near the V/F ring of the inner lumen of the receptor, while buprenorphine and naltrindole bound at the quinacrine site of the receptor. Conclusions The data demonstrate that tandem affinity chromatography – molecular modeling methodology can be used for the rapid assessment of pharmacological activity on nAChRs and to determine the binding site(s) of non‐competitive inhibitors of this receptor.