McMillan Caitlin, Druschitz Amy, Rumbelow Stephen, Borah Ankita, Macfarlane Rebecca, Rattray Zahra, Perrie Yvonne
Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, UK.
Croda International Plc and Avanti Polar Lipids, Alabaster, AL, USA.
J Control Release. 2025 Jul 20;386:114056. doi: 10.1016/j.jconrel.2025.114056.
Lipid nanoparticles (LNPs) have emerged as a vital delivery system for nucleic acids, particularly in the context of mRNA vaccines and therapeutics. This study evaluates the impact of various proprietary ionisable lipids on the physicochemical characteristics, encapsulation efficiency, and expression of mRNA within LNP formulations. We compared 11 novel ionisable lipids against the clinically used ALC-0315 in both in vitro and in vivo models. Alongside comprehensive physicochemical characterisation (including size, zeta potential, and polydispersity index), we uncovered significant variations in vitro in formulation performance influenced by lipid structure, where LNPs formulated with cone-shaped ionisable lipids exhibited markedly higher mRNA expression in HeLa cells compared to the control. In vivo assessments revealed distinct biodistribution patterns, with ALC-0315 formulations demonstrating preferential delivery to the liver, while alternative ionisable lipids shifted distribution toward the spleen, emphasising the role of lipid composition in therapeutic efficacy. Notably, some proprietary LNPs performed well in vitro but showed poor in vivo expression, especially via IV administration, underscoring the importance of delivery context and offering novel insight into route-independent formulation performance trends. However, the overall performance ranking was consistent across both in vivo administration routes, with the best- and worst-performing formulations maintaining their relative expression profiles. Our findings also indicate that while differences in sterol choice modulate LNP properties, the choice of ionisable lipids is crucial for optimising mRNA delivery. Furthermore, the study highlights the discrepancies between in vitro and in vivo results, emphasising the need for further research into the biological complexities of LNP behaviour. Collectively, this work offers new insight into structure-function relationships within LNP systems and provides valuable formulation strategies for next-generation RNA therapies.
脂质纳米颗粒（LNPs）已成为核酸的重要递送系统，特别是在mRNA疫苗和治疗药物领域。本研究评估了各种专利可电离脂质对LNP制剂的物理化学特性、包封效率和mRNA表达的影响。我们在体外和体内模型中，将11种新型可电离脂质与临床使用的ALC-0315进行了比较。除了全面的物理化学表征（包括粒径、zeta电位和多分散指数）外，我们还发现脂质结构会影响体外制剂性能，其中与对照组相比，用锥形可电离脂质配制的LNPs在HeLa细胞中表现出明显更高的mRNA表达。体内评估揭示了不同的生物分布模式，ALC-0315制剂表现出优先向肝脏递送，而其他可电离脂质则使分布转向脾脏，这突出了脂质组成对治疗效果的作用。值得注意的是，一些专利LNP在体外表现良好，但在体内表达不佳，尤其是通过静脉注射给药时，这凸显了递送环境的重要性，并为与给药途径无关的制剂性能趋势提供了新的见解。然而，两种体内给药途径的总体性能排名是一致的，表现最佳和最差的制剂保持其相对表达谱。我们的研究结果还表明，虽然甾醇选择的差异会调节LNP的性质，但可电离脂质的选择对于优化mRNA递送至关重要。此外，该研究突出了体外和体内结果之间的差异，强调需要进一步研究LNP行为的生物学复杂性。总的来说，这项工作为LNP系统内的结构-功能关系提供了新的见解，并为下一代RNA疗法提供了有价值的制剂策略。
