Hydrophobized dextran-spermine conjugate as potential vector for in vitro gene transfection.
Dextran polysaccharide was grafted by reductive-amination with mixtures of spermine and other natural/synthetic oligoamines of two to four amine groups. The transfection efficiencies of the polycations thus obtained were assessed in various cell lines, and found to depend on the spermine contents. Higher spermine ratios of grafted oligoamines resulted in high gene expression, whereas low to negligible expressions were obtained with lower spermine contents. The effect was explained by spermine residues which exhibit altered buffering capacity in comparison to other substituted oligoamines. Hydrophobization of dextran-spermine (D-SPM) was achieved by treating the polymer with N-hydroxysuccinimide derivatives of cholesterol and fatty acids in a mixture of water/THF. The degree of hydrophobization was in the range of 1-30% mol/mol (hydrophobic moieties/primary amine) and the coupling yields were >95% as determined by (1)H-NMR. The oleate-modified D-SPM remarkably enhanced the gene expression in serum rich media, in marked contrast to unmodified D-SPM which resulted with a drastic decrease in the transfection yields. Modified D-SPM derivatives of other fatty acids and cholesterol showed improved transfection yields in comparison to unmodified D-SPM, but to a lower extent when compared to oleate modification. The improvement in cell transfection was attributed to oleate residues which probably play a role in increasing stability and uptake of polycation-DNA complexes.
Azzam
Tony
T
Department of Medicinal Chemistry and Natural Products, Faculty of Medicine, School of Pharmacy, Hebrew University of Jerusalem 91120, Israel.
Eliyahu
Hagit
H
Makovitzki
Arik
A
Linial
Michal
M
Domb
Abraham J
AJ
D000588:Amines / Q000737:chemistry; D000818:Animals; D002240:Carbohydrate Sequence; D002784:Cholesterol / Q000737:chemistry; D004247:DNA / Q000008:administration & dosage; D003911:Dextrans / Q000737:chemistry*; D004337:Drug Carriers / Q000737:chemistry*; D005227:Fatty Acids / Q000737:chemistry; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D013096:Spermine / Q000737:chemistry*; D013388:Succinimides / Q000737:chemistry; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D000588:Amines; D003911:Dextrans; D004337:Drug Carriers; D005227:Fatty Acids; D013388:Succinimides; D049452:Green Fluorescent Proteins; D013096:Spermine; D004247:DNA; D002784:Cholesterol; D008156:Luciferases; D001616:beta-Galactosidase; C001426:N-hydroxysuccinimide