Alpha-selenoconotoxins, a new class of potent alpha7 neuronal nicotinic receptor antagonists.
Disulfide bonds are important structural motifs that play an essential role in maintaining the conformational stability of many bioactive peptides. Of particular importance are the conotoxins, which selectively target a wide range of ion channels that are implicated in numerous disease states. Despite the enormous potential of conotoxins as therapeutics, their multiple disulfide bond frameworks are inherently unstable under reducing conditions. Reduction or scrambling by thiol-containing molecules such as glutathione or serum albumin in intracellular or extracellular environments such as blood plasma can decrease their effectiveness as drugs. To address this issue, we describe a new class of selenoconotoxins where cysteine residues are replaced by selenocysteine to form isosteric and nonreducible diselenide bonds. Three isoforms of alpha-conotoxin ImI were synthesized by t-butoxycarbonyl chemistry with systematic replacement of one ([Sec(2,8)]ImI or [Sec(3,12)]ImI), or both ([Sec(2,3,8,12)]ImI) disulfide bonds with a diselenide bond. Each analogue demonstrated remarkable stability to reduction or scrambling under a range of chemical and biological reducing conditions. Three-dimensional structural characterization by NMR and CD spectroscopy indicates conformational preferences that are very similar to those of native ImI, suggesting fully isomorphic structures. Additionally, full bioactivity was retained at the alpha7 nicotinic acetylcholine receptor, with each selenoanalogue exhibiting a dose-response curve that overlaps with wild-type ImI, thus further supporting an isomorphic structure. These results demonstrate that selenoconotoxins can be used as highly stable scaffolds for the design of new drugs.
Armishaw
Christopher J
CJ
Institute for Molecular Bioscience, University of Queensland, Brisbane, Queensland 4072, Australia.
Daly
Norelle L
NL
Nevin
Simon T
ST
Adams
David J
DJ
Craik
David J
DJ
Alewood
Paul F
PF
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D020916:Conotoxins / Q000737:chemistry / Q000378:metabolism*; D004220:Disulfides / Q000737:chemistry; D015195:Drug Design; D004594:Electrophysiology; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D008969:Molecular Sequence Data; D018733:Nicotinic Antagonists / Q000494:pharmacology*; D009865:Oocytes / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D011978:Receptors, Nicotinic / Q000378:metabolism*; D018036:Selenium Compounds / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000737:chemistry; D014981:Xenopus; D064569:alpha7 Nicotinic Acetylcholine Receptor
C579167:Chrna7 protein, human; D020916:Conotoxins; D004220:Disulfides; D018733:Nicotinic Antagonists; D010455:Peptides; D011978:Receptors, Nicotinic; D018036:Selenium Compounds; D013438:Sulfhydryl Compounds; D064569:alpha7 Nicotinic Acetylcholine Receptor; C037341:sodium selenide