Effects of PEGylation on the preservation of cationic lipid/DNA complexes during freeze-thawing and lyophilization.

摘要

The incorporation of components with covalently attached polyethylene glycol (PEG) into nonviral vectors has been shown to prevent aggregation in serum and extend the circulating half-life of lipid/DNA complexes (lipoplexes) in vivo. The tendency of synthetic vectors to aggregate during processing and storage also represents a significant obstacle in the development of lipoplexes as marketable pharmaceutical products. The extreme instability of lipoplexes formulated as aqueous suspensions has generated interest in preserving nonviral vectors as frozen or lyophilized formulations. Previous work has demonstrated that stabilizing excipients are capable of protecting lipoplexes during freezing and lyophilization, but there is little known about the ability of PEGylation to protect vectors during these stresses. This study incorporates up to 10% by weight dioleoyl phosphatidylethanolamine conjugated to PEG-2000 and PEG-5000 into lipoplexes and assesses the maintenance of particle size and transfection after agitation, freeze-thawing, and lyophilization. Our results indicate that the incorporation of PEGylated components alone (up to 10% by weight) is insufficient to preserve particle size during these stresses. However, when sucrose was employed in combination with PEGylated components, a small protective effect of PEGylation was observed.