Shire Pharmaceuticals, Lexington, Massachusetts 02421, United States.
Nano Lett. 2015 Nov 11;15(11):7300-6. doi: 10.1021/acs.nanolett.5b02497. Epub 2015 Oct 20.
Intracellular delivery of messenger RNA (mRNA) has the potential to induce protein production for many therapeutic applications. Although lipid nanoparticles have shown considerable promise for the delivery of small interfering RNAs (siRNA), their utility as agents for mRNA delivery has only recently been investigated. The most common siRNA formulations contain four components: an amine-containing lipid or lipid-like material, phospholipid, cholesterol, and lipid-anchored polyethylene glycol, the relative ratios of which can have profound effects on the formulation potency. Here, we develop a generalized strategy to optimize lipid nanoparticle formulations for mRNA delivery to the liver in vivo using Design of Experiment (DOE) methodologies including Definitive Screening and Fractional Factorial Designs. By simultaneously varying lipid ratios and structures, we developed an optimized formulation which increased the potency of erythropoietin-mRNA-loaded C12-200 lipid nanoparticles 7-fold relative to formulations previously used for siRNA delivery. Key features of this optimized formulation were the incorporation of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and increased ionizable lipid:mRNA weight ratios. Interestingly, the optimized lipid nanoparticle formulation did not improve siRNA delivery, indicating differences in optimized formulation parameter design spaces for siRNA and mRNA. We believe the general method described here can accelerate in vivo screening and optimization of nanoparticle formulations with large multidimensional design spaces.
细胞内信使 RNA（mRNA）的递送有可能为许多治疗应用诱导蛋白质的产生。尽管脂质纳米粒已显示出在递送小干扰 RNA（siRNA）方面具有相当大的潜力，但它们作为 mRNA 递送剂的用途最近才得到研究。最常见的 siRNA 制剂包含四种成分：含胺的脂质或类脂质材料、磷脂、胆固醇和脂质锚定的聚乙二醇，其相对比例对制剂效力有深远影响。在这里，我们使用实验设计（DoE）方法，包括明确筛选和部分因子设计，开发了一种通用策略，用于优化用于体内肝脏 mRNA 递送的脂质纳米粒制剂。通过同时改变脂质比例和结构，我们开发了一种优化的制剂，与以前用于 siRNA 递送的制剂相比，载有促红细胞生成素-mRNA 的 C12-200 脂质纳米粒的效力提高了 7 倍。该优化制剂的关键特征是 1,2-二油酰基-sn-甘油-3-磷酸乙醇胺（DOPE）的掺入和增加的可离子化脂质：mRNA 重量比。有趣的是，优化的脂质纳米粒制剂并未改善 siRNA 的递送，表明 siRNA 和 mRNA 的优化制剂参数设计空间存在差异。我们相信，这里描述的通用方法可以加速具有大型多维设计空间的纳米粒制剂的体内筛选和优化。
