Varying the chain length in N4,N9-diacyl spermines: non-viral lipopolyamine vectors for efficient plasmid DNA formulation.
The aims of this work are to study the effect of varying the chain length in synthesized N4,N9-diacyl spermines on DNA condensation and then to compare their transfection efficiencies in cell lines. The five novel N4,N9-diacyl lipopolyamines: N4,N9-[didecanoyl, dilauroyl, dimyristoyl, dimyristoleoyl, and dipalmitoyl]-1,12-diamino-4,9-diazadodecane were synthesized from the naturally occurring polyamine spermine. The abilities of these novel compounds to condense DNA and to form nanoparticles were studied using ethidium bromide fluorescence quenching and nanoparticle characterization techniques. Transfection efficiency was studied in FEK4 primary skin cells and in an immortalized cancer cell line (HtTA), and compared with a saturated (distearoyl) analogue and also with the non-liposomal transfection formulation Lipogen, N4,N9-dioleoyl-1,12-diamino-4,9-diazadodecane. By incorporating two aliphatic chains and changing their length in a stepwise manner, we show efficient circular plasmid DNA (pEGFP) formulation and transfection of primary skin and cancer cell lines. Two C14 chains (both saturated or both cis-monounsaturated) were efficient transfecting agents, even in the presence of serum, but they were too toxic. N4,N9-Dioleoyl spermine efficiently condenses pDNA and achieves the highest transfection levels with the highest cell viability among the studied lipopolyamines in cultured cells even in the presence of serum.
Department of Pharmacy and Pharmacology, University of Bath, Bath BA2 7AY, UK.
Hassan M
HM
Ghonaim
Osama A A
OA
Ahmed
Charareh
C
Pourzand
Ian S
IS
Blagbrough
D002460:Cell Line; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002626:Chemistry, Pharmaceutical; D004270:DNA, Circular / Q000235:genetics*; D004304:Dosage Forms; D004996:Ethidium / Q000737:chemistry; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005784:Gene Amplification; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D008081:Liposomes; D053758:Nanoparticles; D010316:Particle Size; D010957:Plasmids; D011073:Polyamines / Q000737:chemistry; D012867:Skin / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D013096:Spermine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism* / Q000633:toxicity; D014162:Transfection
D004270:DNA, Circular; D004304:Dosage Forms; D008081:Liposomes; D011073:Polyamines; D049452:Green Fluorescent Proteins; D013096:Spermine; D004996:Ethidium