Synthetic glycopeptide-based delivery systems for systemic gene targeting to hepatocytes.

摘要: Purpose. To design, synthesize, and test synthetic glycopeptide-baseddelivery systems for gene targeting to hepatocytes by systemicadministration.Methods. All peptides were synthesized the solid phase methoddeveloped using Fmoc chemistry on a peptide synthesizer. The bindingof galactosylated HepG2 cells accessibility of thegalactose residues particle surface was demonstrated acompetition assay 125I-labeleld asialoorosomucoid RCA lectinagglutination assay, respectively. DNA plasmid encoding chloramphenicolacetyl transferase (CAT) complexed with tri-galactosylatedpeptide (GM245.3) or tri-galactosylated lipopeptide (GM246.3) in thepresence an endosomolytic (GM225.1) endosomolyticlipopeptide (GM227.3) obtain particles 100–150 nm insize. plasmid/peptide complexes added cell culturesor intravenously administered tail vein injection into normal miceor rats. Plasmid uptake expression quantified qPCR andELISA, respectively.Results. Multiple antennary glycopeptides that have ability tocondense deliver andcomplexed colloidally stable DNA/peptidecomplexes. Addition DNA/GM245.3/GM225.1 complexes(1:3:1 (−/+/−)) cultures yielded CAT intransfected cells. transfection efficiency significantly reducedin absence galactose ligand removal peptide.Intravenous administration DNA/GM245.3 (1:0.5(−/+)) rats theliver. Substitution GM245.3 GM246.3resulted more particles, 10-fold enhancement inliver uptake. detectable liver followingintravenous DNA/GM246.3 complexes. ofendosomolytic GM227.3 complexes(DNA/GM246.3/GM227.3 (1:0.5:1 (−/+/−))) 5-fold increase inCAT expression. Liver 8-fold 40-fold higher thanlung spleen, respectively, localized only.The enhanced increasing DNAdose volume. liverusing DNA/GM246.3/GM227.3 100-200-fold higherthan formulated glucose. Tissue examination serumbiochemistry did not show any adverse effect after intravenous delivery.Conclusions. Gene achieved systemicadministration well-tolerated system. this glycopeptide deliverysystem dependent structure, activity,colloidal stability,