Introduction of a disulfide bond into a cationic lipid enhances transgene expression of plasmid DNA.
We have introduced a convenient method of synthesis for disulfide-containing cationic lipids. The lipid, 1,2-dioleoyl-sn-glycero-3-succinyl-2-hydroxyethyl disulfide ornithine conjugate (DOGSDSO), was synthesized and used to prepare liposomes in combination with DOPE. The rationale behind the selection of the disulfide bond was to produce a lipid which could be selectively destabilized within the cytosol of the cell. The disulfide bond of DOGSDSO was shown to be cleaved by reductive media leading to destabilization of the liposome/DNA complex, thus increasing the release of pDNA compared to a non-disulfide-containing analog. The introduction of a disulfide bond increases the transfection activity using model animal cell lines. The transfection activity and toxicity of DOTAP, DOGSDSO and its analog in three cell lines were compared. The amount of transgene (e.g. luciferase) produced with the use of DOGSDSO/DOPE was greater than that of DOTAP/DOPE and up to 50 times more than that of its non-disulfide analog. The results indicate disulfide-containing cationic liposomes may act as excellent vectors for gene transfection.
Tang
F
F
Department of Pharmaceutics, University of Florida, Gainesville 32610, USA.
Hughes
J A
JA
D002412:Cations; D004337:Drug Carriers; D005229:Fatty Acids, Monounsaturated; D008055:Lipids / Q000138:chemical synthesis; D008081:Liposomes; D009952:Ornithine / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D010714:Phosphatidylethanolamines; D010957:Plasmids / Q000235:genetics*; D000644:Quaternary Ammonium Compounds; D013329:Structure-Activity Relationship; D014162:Transfection / Q000379:methods*
C110217:1',2'-dioleoyl-sn-glycero-3'-succinyl-1,6-hexanediol ornithiine conjugate; C110216:1,2-dioleoyl-sn-glycero-3-succinyl-2-hydroxyethyl disulfide ornithine conjugate; D002412:Cations; D004337:Drug Carriers; D005229:Fatty Acids, Monounsaturated; D008055:Lipids; D008081:Liposomes; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; C094877:dioleoyl phosphatidylethanolamine; D009952:Ornithine; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane