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Improved synthesis and characterization of cholesteryl oleate-loaded cationic solid lipid nanoparticles with high transfection efficiency for gene therapy applications

AuthorsSuñé-Pou, Marc; Limeres, María J.; Nofrerias, Isaac; Nardi-Ricart, Anna; Prieto-Sánchez, Silvia; El-Yousfi, Younes; Peréz-Lozano, Pilar; García-Montoya, Encarna; Minarro-Carmona, Montserrat; Ramón Tico, Josep; Hernández-Munaín, Cristina; Suñé, Carlos; Suñé-Negre, Josep Mª
KeywordsCholesteryl oleate
Solid lipid nanoparticles
Storage conditions
Gene therapy
Transfection efficiency
Issue Date1-Aug-2019
CitationColloids and Surfaces - B - Biointerfaces
AbstractThe development of new nanoparticle formulations that are capable of high transfection efficiency without toxicity is essential to provide new tools for gene therapy. However, the issues of complex, poorly reproducible manufacturing methods, and low efficiencies during in vivo testing have prevented translation to the clinic. We have previously reported the use of cholesteryl oleate as a novel excipient for solid lipid nanoparticles (SLNs) for the development of highly efficient and nontoxic nucleic acid delivery carriers. Here, we performed an extensive characterization of this novel formulation to make the scale up under Good Manufacturing Practice (GMP) possible. We also describe the complete physicochemical and biological characterization of cholesteryl oleate-loaded SLNs to ensure the reproducibility of this formula and the preservation of its characteristics before and after the lyophilization process. We defined the best manufacturing method and studied the influence of some parameters on the obtained nanoparticles using the Quality by Design (ICH Q8) guideline to obtain cholesteryl oleate-loaded SLNs that remain stable during storage and guarantee in vitro nucleic acid delivery efficacy. Our results indicate that this improved formulation is suitable for gene therapy with the possibility of scale-up the manufacturing of nanoparticles under GMP conditions.
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