In vivo studies of dialkynoyl analogues of DOTAP demonstrate improved gene transfer efficiency of cationic liposomes in mouse lung.
A novel set of dialkynoyl analogues of the cationic, gene delivery lipid DOTAP (1) was synthesized. Structure-activity studies demonstrate that replacement of the cis-double bonds of DOTAP with triple bonds in varying positions alters both the physical properties of the resultant cationic liposome-DNA complexes and their biological functionalities, both in vitro and in vivo. Particularly, in vivo studies demonstrate that pDNA transfection of mouse lung endothelial cells with lead analogue DS(14-yne)TAP (4):cholesterol lipoplexes exhibits double the transfection level with less associated toxicity relative to the well-established DOTAP:cholesterol system. In fact, 4:cholesterol delivers up to 3 times the dose of pDNA in mice than can be tolerated by DOTAP, leading to nearly 3 times greater marker-gene expression. X-ray diffraction studies suggest that lipoplexes containing analogue 4 display increased stability at physiological temperatures. Our results thus suggest that analogue 4 is a potentially strong candidate for the gene therapy of lung tumors.
Fletcher
Steven
S
Imperial College Genetic Therapies Centre, London, UK.
Ahmad
Ayesha
A
Perouzel
Eric
E
Heron
Andrew
A
Miller
Andrew D
AD
Jorgensen
Michael R
MR
D000818:Animals; D019556:COS Cells; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002522:Chlorocebus aethiops; D002784:Cholesterol / Q000494:pharmacology; D004247:DNA / Q000187:drug effects / Q000235:genetics* / Q000494:pharmacology*; D042783:Endothelial Cells / Q000737:chemistry / Q000187:drug effects; D005229:Fatty Acids, Monounsaturated / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D066298:In Vitro Techniques; D008081:Liposomes; D008168:Lung / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D015394:Molecular Structure; D000644:Quaternary Ammonium Compounds / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D014961:X-Ray Diffraction
D005229:Fatty Acids, Monounsaturated; D008081:Liposomes; D000644:Quaternary Ammonium Compounds; D004247:DNA; D002784:Cholesterol; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane