Department of Chemical and Biological Engineering, Monash University, Clayton, VIC 3800, Australia; Monash Suzhou Research Institute, Suzhou 215000, China.
Suzhou CureMed Biopharma Technology Co., Ltd., Suzhou 215000, China.
Int J Pharm. 2023 Jun 10;640:123050. doi: 10.1016/j.ijpharm.2023.123050. Epub 2023 May 16.
Lipid nanoparticles (LNPs) have demonstrated efficacy and safety for mRNA vaccine administration by intramuscular injection; however, the pulmonary delivery of mRNA encapsulated LNPs remains challenging. The atomization process of LNPs will cause shear stress due to dispersed air, air jets, ultrasonication, vibrating mesh etc., leading to the agglomeration or leakage of LNPs, which can be detrimental to transcellular transport and endosomal escape. In this study, the LNP formulation, atomization methods and buffer system were optimized to maintain the LNP stability and mRNA efficiency during the atomization process. Firstly, a suitable LNP formulation for atomization was optimized based on the in vitro results, and the optimized LNP formulation was AX4, DSPC, cholesterol and DMG-PEG at a 35/16/46.5/2.5 (%) molar ratio. Subsequently, different atomization methods were compared to find the most suitable method to deliver mRNA-LNP solution. Soft mist inhaler (SMI) was found to be the best for pulmonary delivery of mRNA encapsulated LNPs. The physico-chemical properties such as size and entrapment efficiency (EE) of the LNPs were further improved by adjusting the buffer system with trehalose. Lastly, the in vivo fluorescence imaging of mice demonstrated that SMI with proper LNPs design and buffer system hold promise for inhaled mRNA-LNP therapies.
脂质纳米颗粒(LNPs)已被证明可通过肌肉注射用于 mRNA 疫苗的给药,具有疗效和安全性;然而,mRNA 包封的 LNPs 的肺部递送至目前仍然具有挑战性。由于分散的空气、空气射流、超声、振动网等的存在,LNPs 的雾化过程会产生剪切应力,导致 LNPs 的聚集或泄漏,这可能不利于细胞间转运和内体逃逸。在本研究中,对 LNP 配方、雾化方法和缓冲系统进行了优化,以在雾化过程中保持 LNP 的稳定性和 mRNA 的效率。首先,基于体外结果优化了适合雾化的 LNP 配方,优化后的 LNP 配方为 AX4、DSPC、胆固醇和 DMG-PEG,摩尔比为 35/16/46.5/2.5(%)。随后,比较了不同的雾化方法,以找到最适合输送 mRNA-LNP 溶液的方法。发现软雾吸入器(SMI)最适合用于肺部递送包裹在 LNPs 中的 mRNA。通过用海藻糖调整缓冲系统,进一步改善了 LNPs 的物理化学性质,如粒径和包封效率(EE)。最后,体内荧光成像实验表明,适当的 LNPs 设计和缓冲系统的 SMI 为吸入型 mRNA-LNP 疗法提供了希望。
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