title: Virtual screening against acetylcholine binding protein.
abstract: The nicotinic acetylcholine receptors (nAChRs) are a member of the ligand-gated ion channel family and play a key role in the transfer of information across neurological networks. The X-ray crystal structure of agonist-bound α(7) acetylcholine binding protein (AChBP) has been recognized as the most appropriate template to model the ligand-binding domain of nAChR for studying the molecular mechanism of the receptor-ligand interactions. Virtual screening of the National Cancer Institute diversity set, a library of 1990 compounds with nonredundant pharmacophore profiles, using AutoDock against AChBPs revealed 51 potential candidates. In vitro radioligand competition assays using [(3)H] epibatidine against the AChBPs from the freshwater snails, Lymnaea stagnalis, and from the marine species, Aplysia californica and the mutant (AcY55W), revealed seven compounds from the list of candidates that had micromolar to nanomolar affinities for the AChBPs. Further investigation on α(7)nAChR expressing in Xenopus oocytes and on the recombinant receptors with fluorescence resonance energy transfer (FRET)-based calcium sensor expressing in HEK cells showed that seven compounds were antagonists of α(7)nAChR, only one compound (NSC34352) demonstrated partial agonistic effect at low dose (10 µM), and two compounds (NSC36369 and NSC34352) were selective antagonists on α(7)nAchR with moderate potency. These hits serve as novel templates/scaffolds for development of more potent and specific in the AChR systems.
authors.affiliation: School of Pharmaceutical Sciences, University of Phayao, Phayao, Thailand.
authors.forename: Maleeruk
authors.initials: M
authors.lastname: Utsintong
authors.forename: Piyanuch
authors.initials: P
authors.lastname: Rojsanga
authors.forename: Kwok-Yiu
authors.initials: KY
authors.lastname: Ho
authors.forename: Todd T
authors.initials: TT
authors.lastname: Talley
authors.forename: Arthur J
authors.initials: AJ
authors.lastname: Olson
authors.forename: Kinzo
authors.initials: K
authors.lastname: Matsumoto
authors.forename: Opa
authors.initials: O
authors.lastname: Vajragupta
mesh_terms: D000465:Algorithms; D000818:Animals; D001048:Aplysia; D001665:Binding Sites; D002352:Carrier Proteins / Q000819:agonists* / Q000037:antagonists & inhibitors*; D018360:Crystallography, X-Ray; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D031541:Fluorescence Resonance Energy Transfer; D057809:HEK293 Cells; D006801:Humans; D058446:Ligand-Gated Ion Channels; D008195:Lymnaea; D009865:Oocytes; D011487:Protein Conformation; D011978:Receptors, Nicotinic / Q000737:chemistry / Q000378:metabolism*; D012984:Software; D014981:Xenopus
chemical_list: C428944:AChBP protein, Lymnaea; D002352:Carrier Proteins; D058446:Ligand-Gated Ion Channels; D011978:Receptors, Nicotinic