title: High-throughput synthesis of conopeptides: a safety-catch linker approach enabling disulfide formation in 96-well format.
abstract: Conotoxins exhibit a high degree of selectivity and potency for a range of pharmacologically relevant targets. The rapid access to libraries of conotoxin analogues, containing multiple intramolecular disulfide bridges for use in drug development, can be a very labor intensive, multi-step task. This work describes a high-throughput method for the synthesis of cystine-bridged conopeptides. Peptides were assembled on a peptide synthesizer employing the Fmoc solid-phase strategy using a safety-catch amide linker (SCAL). Side-chain protecting groups were removed on solid phase before SCAL activation with ammonium iodide in TFA, finally releasing the peptide into the TFA solution. Disulfide bond formation was performed in the cleavage mixture employing DMSO. This improved method allows mixtures of oxidized peptides to be obtained in parallel directly from a peptide synthesizer. A single HPLC purification of the resulting crude oxidized material produced peptides of > 95% purity.
authors.affiliation: Xenome Ltd, 120 Meiers Road, Indooroopilly 4068, Australia. andreas.brust@xenome.com
authors.forename: Andreas
authors.initials: A
authors.lastname: Brust
authors.forename: Alice E
authors.initials: AE
authors.lastname: Tickle
mesh_terms: D000180:Acrylic Resins; D000595:Amino Acid Sequence; D000700:Analgesics / Q000627:therapeutic use / Q000633:toxicity; D000818:Animals; D020916:Conotoxins / Q000138:chemical synthesis* / Q000737:chemistry* / Q000494:pharmacology; D003553:Cystine / Q000737:chemistry; D004220:Disulfides / Q000032:analysis*; D006801:Humans; D008969:Molecular Sequence Data; D050484:Norepinephrine Plasma Membrane Transport Proteins / Q000187:drug effects
chemical_list: D000180:Acrylic Resins; D000700:Analgesics; D020916:Conotoxins; D004220:Disulfides; D050484:Norepinephrine Plasma Membrane Transport Proteins; D003553:Cystine