Characterization of a new class of DNA delivery complexes formed by the local anesthetic bupivacaine

Abstract

Bupivacaine, a local anesthetic and cationic amphiphile, forms stable liposomal-like structures upon direct mixing with plasmid DNA in aqueous solutions. These structures are on the order of 50-70 nm as determined by scanning electron microscopy, and are homogeneous populations as analyzed by density gradient centrifugation. The DNA within these structures is protected from nuclease degradation and UV-induced damage in vitro. Bupivacaine:DNA complexes have a negative zeta potential (surface charge), homogeneous nature, and an ability to rapidly assemble in aqueous solutions. Bupivacaine:DNA complexes, as well as similar complexes of DNA with other local anesthetics, have the potential to be a novel class of DNA delivery agents for gene therapy and DNA vaccines.

Anti-caries DNA vaccine-induced secretory immunoglobulin A antibodies inhibit formation of Streptococcus mutans biofilms in vitro

“…The plasmids were prepared with a commercial EndoFree Plasmid Mega Kit (Qiagen, Hilden, Germany) and stored at -20 °C. A bupivacaine: DNA complex was prepared by adding bupivacaine hydrochloride to an aqueous DNA solution using a fast-mixing method [19] . The final bupivacaine and DNA concentrations were 0.25% and 1 μg/μL, respectively.…”

Effect of Plasmid DNA Vaccine Design and In Vivo Electroporation on the Resulting Vaccine-Specific Immune Responses in Rhesus Macaques

“…Plasmids were propagated in Escherichia coli, isolated from cells by alkaline lysis, purified by column chromatography, and formulated individually at a concentration of 2.5 mg/ml in isotonic citrate buffer (29.3 mM sodium citrate, 0.67 mM citric acid, 150 mM NaCl, 0.34 mM EDTA, pH 6.4 to 6.7) containing 0.25% bupivacaine to allow for the formation of DNA-bupivacaine complexes (45). Final pDNA preparations were shown to consist of Ͼ90% supercoiled pDNA, and residual endotoxin was shown to be Ͻ30 endotoxin units/mg DNA (data not shown).…”

DNA Vaccines: How Much Have We Accomplished In The Last 25 Years?

“…Muscle pretreatment with bupivacaine several days prior to injection of plasmid DNA results in increased DNA uptake, as evidenced by increased expression at the injection site [122,123]. The complex formed with bupivacaine protects the plasmid DNA from nuclease degradation, and intramuscular immunization with this formulation results in higher immune responses against the encoded antigen [124]. Intranasal or intramuscular administration of a DNA vaccine encoding Streptococcus mutants antigens plus bupivacaine induced antibodies, IFN-γ production and significant reduction in dental caries lesions [125].…”