Synthesis and characterization of N-(N',N'-Dimethyl) propyl succinic mono-cholesteryl mono-amide as gene carrier / 中国药学杂志

ABSTRACT

OBJECTIVE: To synthesize a novel cationic cholesterol derivative, N-(N',N'-dimethyl) propyl succinic mono-cholesteryl mono-amide (DMAPA-CHEMS), and evaluate its feasibility as a none-viral gene vehicle. METHODS: DMAPA-CHEMS and phospholipid were employed to produce liposomes by thin membrane dispersion method, and the morphology of liposomes was observed under transmission electron microscope. The average diameter and Zeta potential were determined by laser particle size analyzer. The combination to and protection for DNA were investigated via agarose gel electrophoresis. The uptake of FITC-labelled oligonucleotides into HepG2 cells was determined by flow cytometry, while the transfection promotion of liposomes for GFP-plasmid DNA was observed by inverted fluorescence microscope. RESULTS: The obtained liposomes showed regular shape and uniform size, with a mean diameter of 94.0 nm and Zeta potential of 24 mV. The liposomes could combine DNA completely and protect DNA from the degradation of DNase I when the charge ratio of cationic cholesterol to DNA was more than 4. The cationic liposomes had low cytotoxicity. Although the uptake efficiency of liposomes made of DMAPA-CHEMS was lower than that of DC-Chol, the gene expression efficiency was higher. CONCLUSION: DMAPA-CHEMS is a novel gene carrier with high transfection efficiency and low cytotoxicity, which has a great potential for application.