Herein, we report the efficient synthesis of high molecular weight polymers (up to 331 kDa) that contain β-cyclodextrin within the polymer backbone and the examination of these structures for pDNA delivery within cultured mammalian cells. Two series of polymers were synthesized, one with variation in oligoethyleneamine stoichiometry, Cd146, Cd244, Cd349, and Cd447 (1-4 oligoethyleneamines in the repeat unit, respectively and similar degree of polymerization, nw = 44-49) and another with variation in polymer length (four ethyleneamines in the repeat unit), Cd427, Cd447, Cd493, and Cd4200 [nw = 27, 47, 93, 200] via the "click reaction". The two series of polymers revealed efficient pDNA binding and compaction through gel electrophoresis, dynamic light scattering, and transmission electron microscopy experiments. The DNase protection assay showed a decrease in pDNA degradation with an increase in the polymer amine stoichiometry, where polymer Cd349 and all of the Cd4 analogs completely protected pDNA for up to 8 h in serum. The cellular uptake and gene expression profiles were examined in HeLa cells, which similarly demonstrated that both the series of polymers had high pDNA delivery where, Cd349 and Cd493 had the most effective luciferase gene expression. In addition, the cell viability profiles were quite high with all of the structures.
|Original language||English (US)|
|Number of pages||11|
|State||Published - Feb 2009|
Bibliographical noteFunding Information:
The authors gratefully acknowledge Dr. Weisen Wong (Viscotek) for his help in the development of new cation exchange column for the analysis of the cyclodextrin polymers using GPC. The funding of this work was provided in part by the NSF CAREER Award program (CHE-0449774) and the Alfred P. Sloan Foundation.