Symmetrical and asymmetric dimeric cationic amphiphiles based on lipopeptides of irregular structure as potential components of cationic liposomes

Objectives. Gene therapy techniques based on the introduction of therapeutic nucleic acids into body cells are currently being developed for the treatment of diseases with a genetic etiology. Among modern drug delivery systems, nonviral agents based on the use of a variety of lipids to produce liposomes and micelles occupy a special place. This work sets out to synthesize and study the properties of dimeric cationic amphiphiles of irregular structure with symmetric and asymmetric hydrophobic blocks in order to determine the influence of structure on physicochemical properties and evaluate the prospects of their application as transfection agents.

Methods. The formation of hydrophobic and hydrophilic blocks involves reactions of L-cystine derivatives and L-glutamic acid and diethanolamine diesters using the condensing agents: dicyclohexylcarbodiimide (DCC) + 4-(dimethylamino)pyridine (DMAP) or hexafluorophosphate benzotriazole tetramethyl uranium (HBTU) + diisopropylethylamine (DIPEA). In order to isolate the reaction products from the reaction mixture, column chromatography and/or preparative thin-layer chromatography on silica gel were used. The structure of the obtained compounds was confirmed by 1H nuclear magnetic resonance spectroscopy and mass spectrometry. Synthesized lipopeptides in aqueous medium formed liposomal dispersions whose particle size was determined by photon correlation spectroscopy.

Results. Schemes for the preparation of novel dimeric cationic amphiphiles based on L-cystine derivatives were devised. The hydrophobic blocks of the obtained compounds include diesters of diethanolamine and L-glutamic acid (C10, C14, and C16). Targeted lipopeptides were used to obtain liposomal dispersed systems mixed with natural lipids. The hydrodynamic size of the particles formed in all dispersions was determined to be within the range of 50 to 200 nm.

Conclusions. The physicochemical properties of aqueous dispersions based on the synthesized compounds were investigated. Dimeric amphiphiles mixed with phosphatidylcholine and cholesterol form liposomal particles. The impact of amphiphile structure on aggregate size was demonstrated. The number of L-ornithine residues (0, 1, 2) in the target products was found to be the most significant parameter affecting the particle size.

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