University of Pennsylvania, Perelman School of Medicine, Department of Medicine, Philadelphia, PA, USA.
The University of Texas at Austin, College of Pharmacy, Division of Molecular Pharmaceutics and Drug Delivery, Austin, TX, USA.
J Control Release. 2024 Nov;375:829-838. doi: 10.1016/j.jconrel.2024.09.030. Epub 2024 Oct 10.
Messenger RNA (mRNA) vaccines have revolutionized the fight against infectious diseases and are poised to transform other therapeutic areas. Lipid nanoparticles (LNP) represent the most successful delivery system for mRNA. While the mRNA-LNP products currently in clinics are stored as frozen suspensions, there is evidence that freeze-drying mRNA-LNP into dry powders can potentially enable their storage and handling at non-freezing temperatures. Previously, we successfully applied thin-film freeze-drying (TFFD) to transform a polyadenylic acid [poly(A)]-LNP formulation from a liquid suspension to dry powders. The poly(A)-LNP were structurally multilamellar spheres without blebs, but the mRNA vaccines in clinics are comprised of mRNA-LNP that are structurally spheres surrounded by a unilamellar lipid bilayer, with some containing blebs, and it was reported that the presence of blebs increases the sensitivity of mRNA-LNP to freeze-drying-induced stress. In the present study, using an influenza A virus hemagglutinin (HA) mRNA in LNP that were structurally similar to that in the COVID-19 mRNA vaccines currently in clinic, we studied the effect of TFFD on the physical properties, internal structure, as well as immunogenicity of the HA mRNA-LNP vaccine. We concluded that TFFD can be utilized to prepare dry powders of the HA mRNA-LNP, but a sufficient amount of excipients were needed to minimize changes in the physical properties, structure, and immunogenicity of the HA mRNA-LNP vaccine.
信使 RNA(mRNA)疫苗改变了传染病的防治方式,并有望改变其他治疗领域。脂质纳米粒(LNP)是 mRNA 的最成功的递送系统。虽然目前临床应用的 mRNA-LNP 产品是以冷冻悬浮液的形式储存的,但有证据表明,将 mRNA-LNP 冻干成干粉可以潜在地实现它们在非冷冻温度下的储存和处理。以前,我们成功地应用了薄膜冷冻干燥(TFFD)将 poly(A)-LNP 制剂从液体悬浮液转化为干粉。poly(A)-LNP 是结构上的多层球体,没有气泡,但临床应用的 mRNA 疫苗由结构上是球体的 mRNA-LNP 组成,球体周围有一层单分子层脂质双层,其中一些含有气泡,据报道气泡的存在增加了 mRNA-LNP 对冻干诱导的应激的敏感性。在本研究中,使用流感 A 病毒血凝素(HA)mRNA 包封在 LNP 中,其结构与目前临床应用的 COVID-19 mRNA 疫苗中的结构相似,我们研究了 TFFD 对 HA mRNA-LNP 疫苗的物理性质、内部结构以及免疫原性的影响。我们得出结论,TFFD 可用于制备 HA mRNA-LNP 的干粉,但需要足够量的赋形剂来最小化 HA mRNA-LNP 疫苗的物理性质、结构和免疫原性的变化。
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