title: Affinities of bispyridinium non-oxime compounds to [(3)H]epibatidine binding sites of Torpedo californica nicotinic acetylcholine receptors depend on linker length.
abstract: The toxicity of organophosphorus nerve agents or pesticides arises from accumulation of acetylcholine and overstimulation of both muscarinic and nicotinic acetylcholine receptors (mAChRs and nAChRs) due to inhibition of acetylcholinesterase (AChE). Standard treatment by administration of atropine and oximes, e.g., obidoxime or pralidoxime, focuses on antagonism of mAChRs and reactivation of AChE, whereas nicotinic malfunction is not directly treated. An alternative approach would be to use nAChR active substances to counteract the effects of accumulated acetylcholine. Promising in vitro and in vivo results were obtained with the bispyridinium compounds SAD-128 (1,1'-oxydimethylene bis(4-tert-butylpyridinium) dichloride) and MB327 (1,1'-(propane-1,3-diyl)bis(4-tert-butylpyridinium) di(iodide)), which were partly attributed to their interaction with nAChRs. In this study, a homologous series of unsubstituted and 4-tert-butyl-substituted bispyridinium compounds with different alkane linker lengths was investigated in competition binding experiments using [(3)H]epibatidine as a reporter ligand. Additionally, the effect of the well-characterised MB327 on the [(3)H]epibatidine equilibrium dissociation (KD) constant in different buffers was determined. This study demonstrated that divalent cations increased the affinity of [(3)H]epibatidine. Since quaternary ammonium molecules are known to inhibit AChE, the obtained affinity constants of the tested bispyridinium compounds were compared with the inhibition of human AChE. In competition experiments, bispyridinium derivatives of longer linker length displaced [(3)H]epibatidine and inhibited AChE strongly. Bispyridinium compounds with short linkers, at most, have an allosteric interaction with the [(3)H]epibatidine binding sites and barely inhibited AChE. In dependence on alkane linker length, the bispyridinium compounds seemed to interact at different binding sites. However, the exact binding sites of the bispyridinium compounds responsible for the positive pharmacological effects have still not been identified, making predictive drug design difficult.
authors.affiliation: Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstrasse 11, 80937 Munich, Germany. Electronic address: KarinNiessen@bundeswehr.org.
authors.forename: K V
authors.initials: KV
authors.lastname: Niessen
authors.forename: T
authors.initials: T
authors.lastname: Seeger
authors.forename: J E H
authors.initials: JE
authors.lastname: Tattersall
authors.forename: C M
authors.initials: CM
authors.lastname: Timperley
authors.forename: M
authors.initials: M
authors.lastname: Bird
authors.forename: C
authors.initials: C
authors.lastname: Green
authors.forename: H
authors.initials: H
authors.lastname: Thiermann
authors.forename: F
authors.initials: F
authors.lastname: Worek
mesh_terms: D000110:Acetylcholinesterase / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000378:metabolism*; D002619:Chemical Warfare Agents / Q000633:toxicity; D002800:Cholinesterase Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D029941:Fish Proteins / Q000378:metabolism; D006801:Humans; D008024:Ligands; D062025:Organophosphate Poisoning / Q000188:drug therapy / Q000378:metabolism; D010575:Pesticides / Q000633:toxicity; D011725:Pyridines / Q000378:metabolism*; D011726:Pyridinium Compounds / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D011978:Receptors, Nicotinic / Q000187:drug effects / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014101:Torpedo / Q000378:metabolism
chemical_list: D019086:Bridged Bicyclo Compounds, Heterocyclic; D002619:Chemical Warfare Agents; D002800:Cholinesterase Inhibitors; D029941:Fish Proteins; D008024:Ligands; C571852:MB327; D010575:Pesticides; D011725:Pyridines; D011726:Pyridinium Compounds; D011978:Receptors, Nicotinic; C016845:SAD-128; D000110:Acetylcholinesterase; C082748:epibatidine
keywords: Acetylcholinesterase; Bispyridinium compounds; Linker length; Nicotinic acetylcholine receptor; Torpedo californica; [(3)H]epibatidine