New poly(d-glucaramidoamine)s induce DNA nanoparticle formation and efficient gene delivery into mammalian cells.
In this report, four new poly(d-glucaramidoamine)s (1-4) have been designed to lower the toxicity of conventional polymeric nucleic acid delivery vehicles by incorporating a carbohydrate comonomer within a polyethylenimine (PEI)-like backbone. Polymers 1-4 were synthesized via polycondensation of esterified d-glucaric acid and four different amine-containing comonomers [diethylenetriamine (1), triethylenetetramine (2), tetraethylenepentamine (3), and pentaethylenehexamine (4)] in methanol. Viscometry and NMR studies suggest that the polymers are mostly linear (for 1-4, the alpha value in the Mark-Houwink-Sakurada equation = 0.6-0.7), thus indicating that polymerization occurs predominantly through the primary amines with a low degree of branching off the secondary amines. Results of gel electrophoresis shift assays show that polymers 1-4 bind pDNA at N/P ratios of 5, 3, 2, and 2, respectively. Also, dynamic light scattering and TEM experiments indicate that 1-4 compact DNA into nanoparticles (polyplexes) between 140 and 440 nm at an N/P ratio of 30. Furthermore, polyplexes formed with 1-4 deliver pDNA (plasmid DNA) containing the firefly luciferase reporter gene to BHK-21 cells in a nontoxic and highly efficient manner (as determined by luciferase gene expression). In particular, polymer 4 reveals very high delivery efficiency (equivalent to linear PEI). This result may be due in part to the "proton sponge" hypothesis proposed by Behr et al. Polymers containing amines that are protonated in the endosomal pH range (between about 7.4-5.0) reveal enhanced gene delivery profiles.
Liu
Yemin
Y
Department of Chemistry, University of Cincinnati, Cincinnati, Ohio 45221-0172, USA.
Wenning
Laura
L
Lynch
Matthew
M
Reineke
Theresa M
TM
D000818:Animals; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D006224:Cricetinae; D004247:DNA / Q000737:chemistry* / Q000235:genetics*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics / Q000494:pharmacology*; D005937:Glucaric Acid / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000494:pharmacology*; D015394:Molecular Structure; D036103:Nanotechnology; D009757:Nylons / Q000138:chemical synthesis / Q000494:pharmacology*; D010316:Particle Size; D010957:Plasmids / Q000235:genetics
D009757:Nylons; C491990:poly(D-glucaramidoamine); D004247:DNA; D005937:Glucaric Acid