Spanedda Maria Vittoria, De Giorgi Marcella, Hassane Fatouma Saïd, Schuber Francis, Bourel-Bonnet Line, Frisch Benoît
Laboratoire de Conception et Applications des Molécules Bioactives, Faculté de Pharmacie, UMR 7199 CNRS/Université de Strasbourg, 74 Route du Rhin, 67401, Illkirch, France.
Methods Mol Biol. 2017;1522:93-106. doi: 10.1007/978-1-4939-6591-5_8.
Click chemistry represents a new bioconjugation strategy that can be used to conveniently attach various ligands to the surface of preformed liposomes. This efficient and chemoselective reaction involves a Cu(I)-catalyzed azide-alkyne cycloaddition which can be performed under mild experimental conditions in aqueous media. Here we describe the application of a model click reaction to the conjugation, in a single step, of unprotected α-1-thiomannosyl ligands, functionalized with an azide group, to liposomes containing a terminal alkyne-functionalized lipid anchor. Excellent coupling yields have been obtained in the presence of bathophenanthroline disulfonate, a water soluble copper-ion chelator, acting as a catalyst. No vesicle leakage is triggered by this conjugation reaction and the coupled mannose ligands are exposed at the surface of the liposomes. The major limitation of Cu(I)-catalyzed click reactions is that this conjugation is restricted to liposomes made of saturated (phospho)lipids. To circumvent that constraint, an example of alternative copper-free azide-alkyne click reaction has been developed. Molecular tools and results are presented here.
点击化学代表了一种新的生物共轭策略，可用于将各种配体方便地连接到预制脂质体的表面。这种高效且具有化学选择性的反应涉及铜（I）催化的叠氮化物-炔烃环加成反应，该反应可在水性介质中的温和实验条件下进行。在此，我们描述了一种模型点击反应在一步中将用叠氮基团功能化的未保护α-1-硫代甘露糖基配体与含有末端炔烃功能化脂质锚定基团的脂质体进行共轭的应用。在水溶性铜离子螯合剂二磺酸邻菲啰啉作为催化剂的情况下，已获得了优异的偶联产率。这种共轭反应不会引发囊泡泄漏，并且偶联的甘露糖配体暴露在脂质体表面。铜（I）催化的点击反应的主要局限性在于这种共轭仅限于由饱和（磷酸）脂质制成的脂质体。为了规避这一限制，已开发出一种无铜叠氮化物-炔烃点击反应的替代示例。本文展示了分子工具和结果。
