Shahsavandi S, Nasr Isfahani H, Hariri A A, Sharifnia Z, Soleimani S, Moradi A
Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization, Karaj, Iran.
Biotechnology Department, Behyaar Sanaat Sepahan Company, Isfahan, Iran.
Arch Razi Inst. 2024 Dec 31;79(6):1217-1226. doi: 10.32592/ARI.2024.79.6.1217. eCollection 2024 Dec.
The COVID-19 pandemic has significantly highlighted the successful application of lipid nanoparticles (LNPs) as an advanced platform for mRNA vaccine delivery. Ionizable lipid is the main component for complexing the mRNA in LNP formulation and delivery. In the first step of this study, we used the native safflower oil seed to prepare dilinoleyl alcohol. Then the cationic lipid DLin-MC3-DMA (MC3) was synthesized by mixing the alcohol with dimethylamino butyric acid. Safflower-derived MC3 was applied to formulate an LNP vector with standard composition. The efficiency of the synthetic cationic lipid was evaluated for delivering an mRNA-based vaccine encoding the receptor-binding domain (RBD) of SARS-CoV-2. The produced mRNA-LNP vaccine candidate was evaluated in size, morphology, mRNA encapsulation efficiency, apparent pKa, and stability for nucleic acid delivery. Cellular uptake was determined by measuring the percentage of GFP expression, and cytotoxicity was assayed using MTT. The MC3 formation was confirmed by the NMR spectra and used as a cationic lipid in LNP formulation. The obtained LNPs had positively charged and appropriate particle sizes (~80 nm) to confer proper encapsulation efficiency for mRNA delivery and stability. The LNPs were shown to be effective in the transfection of mRNA transcripts into HEK293T cells. A high level (72.34%) of cellular uptake was determined by measuring the percentage of GFP expression. The cytotoxicity assay using MTT showed that both LNP and mRNA-LNP were non-toxic to cells. These data demonstrate the potential of the proposed safflower-derived cationic lipid in the formulation of LNP. The carrier provides a promising platform for the efficient delivery of mRNA . Further evaluations of its potential for delivery are needed.
新冠疫情显著凸显了脂质纳米颗粒（LNPs）作为一种先进的mRNA疫苗递送平台的成功应用。可电离脂质是LNP制剂中与mRNA复合并进行递送的主要成分。在本研究的第一步，我们使用天然红花籽制备二亚油醇。然后将该醇与二甲基氨基丁酸混合合成阳离子脂质DLin-MC3-DMA（MC3）。将源自红花的MC3用于配制具有标准组成的LNP载体。评估了这种合成阳离子脂质递送编码严重急性呼吸综合征冠状病毒2（SARS-CoV-2）受体结合域（RBD）的基于mRNA的疫苗的效率。对制备的mRNA-LNP候选疫苗进行了尺寸、形态、mRNA包封效率、表观pKa以及核酸递送稳定性方面的评估。通过测量绿色荧光蛋白（GFP）表达的百分比来确定细胞摄取情况，并使用噻唑蓝（MTT）法测定细胞毒性。通过核磁共振光谱确认了MC3的形成，并将其用作LNP制剂中的阳离子脂质。所获得的LNPs带正电荷且粒径合适（约80 nm），以实现mRNA递送的适当包封效率和稳定性。结果表明，LNPs在将mRNA转录物转染到HEK293T细胞中是有效的。通过测量GFP表达的百分比确定细胞摄取水平较高（72.34%）。使用MTT进行的细胞毒性测定表明，LNP和mRNA-LNP对细胞均无毒。这些数据证明了所提出的源自红花的阳离子脂质在LNP制剂中的潜力。该载体为高效递送mRNA提供了一个有前景的平台。需要进一步评估其递送潜力。
