New Cationic Carbohydrate-Modified Amphiphiles and Liposomes for Effective Delivery of Short Nucleic Acids into Eukaryotic Cells

Objective: The development of systems for targeted delivery of nucleic acids (NAs) is necessary to ensure their selective transport to the site of therapeutic action. The aim of this work was to synthesize carbohydrate-modified amphiphiles containing a spermine residue, required for compaction and binding to NAs, as well as a diglyceride residue for forming lipid aggregates and a carbohydrate residue (lactose or D-mannose) for improving the hydrophilic–lipophilic balance of the molecule. The lactose residue can serve as a targeting ligand for NA delivery into liver hepatocytes, and the D-mannose residue can perform specific NA transport into dendritic cells and macrophages. Methods: New carbohydrate-modified cationic amphiphiles were obtained by organic synthesis, and their aqueous dispersions or cationic liposomes were prepared. Cytotoxicity of the cationic amphiphiles and liposomes was performed using the MTT assay on HEK 293 and BHK cell lines in the absence of fetal bovine serum (FBS). Complexes of the cationic amphiphiles or liposomes with NAs (FITC-ODN, pDNA, and siRNA) were formed at various component ratios (N/P), and the efficiency of transfection in HEK 293 and BHK IR-780 cells was assessed by flow cytometry. Results and Discussion: New cationic amphiphiles containing lactose or D-mannose residues were synthesized. The cationic amphiphiles, whatever the structure of their carbohydrate residue, effectively deliver a short FITC-ODN into HEK293 cells in the presence of FBS, and are nontoxic. The cationic liposome formed by the lactose-containing amphiphile and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) successfully delivers short NAs (FITC-ODN and siRNA) both in the absence and in the presence of serum in the culture media. Conclusions: The obtained carbohydrate-modified cationic amphiphiles, both individually and as component of cationic liposomes, hold promise to be used as systems for the delivery of short nucleic acids in further development of drugs for gene therapy.