Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
2507141 | International Journal of Pharmaceutics | 2006 | 10 Pages |
Abstract
The synthesis and transfection potential of a novel cationic cholesterol cytofectin with a dimethylamino head group and a long 12 atom, 15Â Ã
spacer incorporating relatively polar amido and dicarbonyl hydrazine linkages are reported. Thus N,N-dimethylaminopropylamidosuccinylcholesterylformylhydrazide (MS09) in equimolar admixture with dioleoylphosphatidylethanolamine (DOPE) forms stable unilamellar liposomes (80-150Â nm) which cluster into very effective transfecting, serum nuclease-resistant, lipoplexes with DNA (180-200Â nm) at a liposome+/DNAâ molar charge ratio of 2.8:1 (12:1, w/w). Gel retardation and ethidium displacement assays confirmed that DNA was fully liposome-associated and maximally compacted at this ratio. Transfection levels in three human transformed epithelial cell lines, as established by luciferase transgene activity, was found to be optimal at this charge ratio and in the following order: cervical carcinoma (HeLa)Â >Â oesophageal carcinoma (SNO)Â >Â hepatoblastoma (HepG2). Activity in the murine fibroblast line NIH-3T3 was comparable to that in HepG2 cells. MS09 lipoplexes achieved approximately three-times and two-times greater activity than Lipofectin® complexes in HeLa and SNO cells, respectively, whilst comparable levels were recorded in HepG2 and NIH-3T3 cells. MS09 lipoplexes were well tolerated by HepG2, HeLa and SNO cells with cell numbers found to be 80, 85 and 75% of untreated cultures, respectively, at the optimal transfection concentration. These lipoplexes also exhibited high activity in the presence of 10% foetal bovine serum (FBS) in HeLa (17% inhibition) and HepG2 (33% inhibition) cells.
Keywords
DC-CholNHSDCCPEIHBSBCARLUHEPESN-hydroxysuccinimideDOTAPdicyclohexylcarbodiimidePBSDMFDMSOhepes buffered salineTembicinchoninic acidTransfectionminimum essential mediumdimethylformamidedioleoylphosphatidylethanolamineDimethylsulfoxidePhosphate buffered salineCationic liposomeMEMTransmission electron microscopyrelative light unitspolyethyleneimineDOPE
Related Topics
Health Sciences
Pharmacology, Toxicology and Pharmaceutical Science
Pharmaceutical Science
Authors
Moganavelli Singh, Mario Ariatti,