Low Molecular Weight Dextrans Stabilize Nonviral Vectors During Lyophilization at Low Osmolalities: Concentrating Suspensions by Rehydration to Reduced Volumes

Stabilization of nonviral vectors during freezing and drying requires formulation with protective excipients such that transfection rates and physical characteristics are maintained upon reconstitution. While many studies have demonstrated the ability of disaccharides (e.g., sucrose) to effectively protect nonviral vectors during lyophilization, the sucrose/DNA weight ratios required to achieve stability result in formulations that are not osmotically compatible with the subcutaneous (SC) or intramuscular (IM) injection of a typical dose of plasmid DNA. In an effort to reduce the formulation osmolality, dextrans possessing a range of molecular weights were investigated for their ability to serve as protectants. Dextran 3000 proved to be the most effective of the dextrans tested, and offered similar protection to sucrose on a weight basis. However, the advantage of employing this excipient is that the resulting osmolality is reduced by approximately 40% as compared to an equivalent weight of sucrose. Moreover, the use of dextran allows lyophilized vector preparations to be rehydrated to reduced volumes, essentially concentrating vectors prior to administration. Utilizing a combination of dextran 3000 and sucrose, we demonstrate that complexes of polyethylenimine (PEI) and DNA lyophilized at 0.1 mg/mL can be concentrated tenfold upon rehydration, resulting in an isotonic formulation containing 1 mg/mL DNA that can provide more realistic injection volumes for animal studies, and is compatible with clinical trials involving SC and IM injection. © 2005 Wiley-Liss, Inc. and the American Pharmacists Association