Athaydes Seabra Ferreira Heloísa, Ricardo Aluotto Scalzo Júnior Sérgio, Kelton Santos de Faria Kevin, Henrique Costa Silva Gabriel, Túllio Rodrigues Alves Marco, Oliveira Lobo Anderson, Pires Goulart Guimarães Pedro
Department of Physiology and Biophysics, Institute of Biological Sciences, Federal University of Minas Gerais, Belo Horizonte, 31270-901 Minas Gerais, Brazil.
Department of Materials Engineering, Federal University of Piauí, Teresina, 64049-550, Piauí, Brazil.
Int J Pharm. 2024 Nov 15;665:124696. doi: 10.1016/j.ijpharm.2024.124696. Epub 2024 Sep 10.
Advances in gene therapy, exemplified by mRNA vaccines against COVID-19, highlight the importance of lipid nanoparticles (LNPs) for nucleic acid delivery despite challenging storage conditions. Substituting mRNA with pDNA in LNPs may enhance stability and efficacy, yet maintaining LNP stability poses challenges, particularly during freeze-drying. Cryoprotectants offer potential to mitigate destabilization, improving LNP properties and in vivo performance. Here, we evaluated the effects of different concentrations of various cryoprotectants on the freeze-drying process of pDNA-loaded LNPs, assessing their physicochemical characteristics and transfection efficiency. Stability was examined under various storage conditions, confirming biological efficacy post-storage. Our results highlight the role of cryoprotectants in optimizing freeze-drying for the extended shelf life of nucleic acid-loaded LNPs. Trehalose emerged as an efficient cryoprotectant, maintaining LNP stability after the freeze-drying process for up to 2 years, with diameters and transfection efficiency comparable to fresh formulations. These findings demonstrate the optimized concentration of cryoprotectants to sustain LNP stability despite freeze-drying and prolonged storage, providing valuable insights for nucleic acid-based therapies.
以针对新冠病毒的信使核糖核酸（mRNA）疫苗为代表的基因治疗进展，凸显了脂质纳米颗粒（LNP）在核酸递送方面的重要性，尽管其储存条件具有挑战性。在LNP中用质粒DNA（pDNA）替代mRNA可能会提高稳定性和疗效，但维持LNP的稳定性仍面临挑战，尤其是在冷冻干燥过程中。冷冻保护剂有可能减轻去稳定化作用，改善LNP的性质和体内性能。在此，我们评估了不同浓度的各种冷冻保护剂对负载pDNA的LNP冷冻干燥过程的影响，评估了它们的物理化学特性和转染效率。在各种储存条件下检查稳定性，确认储存后的生物学疗效。我们的结果凸显了冷冻保护剂在优化冷冻干燥以延长负载核酸的LNP保质期方面的作用。海藻糖成为一种有效的冷冻保护剂，在冷冻干燥过程后长达2年的时间里保持LNP的稳定性，其直径和转染效率与新鲜制剂相当。这些发现证明了冷冻保护剂的优化浓度，以在冷冻干燥和长期储存后维持LNP的稳定性，为基于核酸的疗法提供了有价值的见解。
