title: Differential reactivity of maleimide and bromoacetyl functions with thiols: application to the preparation of liposomal diepitope constructs.
abstract: The comparative reactivity of maleimide and bromoacetyl groups with thiols (2-mercaptoethanol, free cysteine, and cysteine residues present at the N-terminus of peptides) was investigated in aqueous media. These studies were performed (i) with water-soluble functionalized model molecules, i.e., polyoxyethylene-based spacer arms that could also be coupled to lipophilic anchors destined to be incorporated into liposomes, and (ii) with small unilamellar liposomes carrying at their surface these thiol-reactive functions. Our results indicate that an important kinetic discrimination (2-3 orders of magnitude in terms of rate constants) can be achieved between the maleimide and bromoacetyl functions when the reactions with thiols are performed at pH 6.5. The bromoacetyl function which reacts at higher pH values (e.g., pH 9.0) retained a high chemoselectivity; i.e., under conditions where it reacted appreciably with the thiols of, e.g., HS-peptides, it did react with other nucleophilic functions such as alpha- and epsilon-amino groups or imidazole, which could also be present in peptides. This differential reactivity was applied to design chemically defined and highly immunogenic liposomal diepitope constructs as synthetic vaccines, i.e., vesicles carrying at their surface two different peptides conjugated each to a specific amphiphilic anchor. This was realized by coupling sequentially at pH 6.5 and 9.0 two HS-peptides to preformed vesicles containing lipophilic anchors functionalized with maleimide and bromoacetyl groups [Boeckler, C., et al. (1999) Eur. J. Immunol. 29, 2297-2308].
authors.affiliation: Laboratoire de Chimie Bioorganique, UMR 7514 CNRS-Université Louis Pasteur, Faculté de Pharmacie, 74 route du Rhin, 67400 Strasbourg-Illkirch, France.
authors.forename: P
authors.initials: P
authors.lastname: Schelté
authors.forename: C
authors.initials: C
authors.lastname: Boeckler
authors.forename: B
authors.initials: B
authors.lastname: Frisch
authors.forename: F
authors.initials: F
authors.lastname: Schuber
mesh_terms: D000085:Acetates / Q000737:chemistry*; D000595:Amino Acid Sequence; D003545:Cysteine / Q000737:chemistry; D004337:Drug Carriers; D000939:Epitopes / Q000737:chemistry* / Q000276:immunology; D006863:Hydrogen-Ion Concentration; D018796:Immunoconjugates / Q000737:chemistry* / Q000276:immunology; D007700:Kinetics; D008081:Liposomes / Q000138:chemical synthesis* / Q000276:immunology; D008301:Maleimides / Q000737:chemistry*; D008623:Mercaptoethanol / Q000737:chemistry; D009842:Oligopeptides / Q000737:chemistry* / Q000276:immunology; D010714:Phosphatidylethanolamines / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000737:chemistry*
chemical_list: D000085:Acetates; D004337:Drug Carriers; D000939:Epitopes; D018796:Immunoconjugates; D008081:Liposomes; D008301:Maleimides; D009842:Oligopeptides; D010714:Phosphatidylethanolamines; D013438:Sulfhydryl Compounds; C043592:maleimide; D008623:Mercaptoethanol; C016274:bromoacetate; D003545:Cysteine