Efficacy increase of lipid nanoparticles in vivo by inclusion of bis(monoacylglycerol)phosphate - Supplementary data

Aim: To investigate the effect of incorporating bis(monoacylglycerol)phosphate (BMP) lipid into a
lipid nanoparticle and the functional transport of mRNA by the formulated nanoparticles in vivo.
Materials & methods: The nanoparticles were prepared from ionizable lipid, 1,2-distearoyl-sn-glycerol-3-
phosphocholine, cholesterol, 1,2-dimyristoyl-sn-glycerol PEG 2000, BMP and formulated mRNA encoding
human erythropoietin. We measured the effect of BMP on physicochemical properties and impact on
functional efficacy to transport mRNA to its target cells/tissue as measured by protein expression both in
vitro and in vivo. Results: Lipid nanoparticles composed of BMP displayed increased endosomalmembrane
fusion and improved mRNA delivery to the cytosol. Conclusion: The results establish the foundation for
future development of these nanoparticulated entities by designing new BMP derivatives and correlating
structures to enhanced pharmacokinetic profiles.

研究目的：探究将双单酰甘油磷酸酯（bis(monoacylglycerol)phosphate，BMP）脂质掺入脂质纳米粒中，以及所配制的纳米粒在体内介导mRNA功能性转运的效果。材料与方法：本研究以可电离脂质、1,2-二硬脂酰-sn-甘油-3-磷酸胆碱（1,2-distearoyl-sn-glycerol-3-phosphocholine）、胆固醇、1,2-二肉豆蔻酰-sn-甘油聚乙二醇2000（1,2-dimyristoyl-sn-glycerol PEG 2000）、BMP为原料制备纳米粒，并配制编码人促红细胞生成素（human erythropoietin）的mRNA。通过体外及体内的蛋白表达水平检测，评估BMP对纳米粒理化性质的影响，以及其将mRNA转运至靶细胞/靶组织的功能效力。研究结果：掺入BMP的脂质纳米粒可增强内体膜融合能力，并提升mRNA向细胞质的递送效率。结论：本研究结果为后续开发此类纳米粒制剂奠定了基础，未来可通过设计新型BMP衍生物，并建立其结构与优化药代动力学特性之间的关联，进一步推动相关技术发展。