Abstract

Functionalized drug delivery systems have been investigated to improve the targetability and intracellular translocation of therapeutic drugs. We developed high functionality and quality lipids that met unique requirements, focusing on the quality of functional lipids for the preparation of targeted nanoparticles using microfluidic devices. While searching for a lipid with high solubility and dispersibility in solvents, which is one of the requirements, we noted that KK-(EK)₄-lipid imparts nonspecific cellular association to polyethylene glycol (PEG)-modified (PEGylated) liposomes, such as cell-penetrating peptides (CPPs). We investigated whether KK-(EK)₄-lipid, which has a near-neutral charge, is a novel CPP-modified lipid that enhances the intracellular translocation of nanoparticles. However, the cellular association mechanism of KK-(EK)₄-lipid is unknown. Therefore, we synthesized (EK)_n-lipid derivatives based on the sequence of KK-(EK)₄-lipid and determined the sequence sites involved in cellular association. In addition, KK-(EK)₄-lipid was applied to extracellular vesicles (EVs) and mRNA encapsulated lipid nanoparticles (mRNA-LNPs). KK-(EK)₄-lipid-modified EVs and mRNA-LNPs showed higher cellular association and in vitro protein expression, respectively, compared to unmodified ones. We elucidated KK-(EK)₄-lipid to have potential for applicability in the intracellular delivery of liposomes, EVs, and mRNA-LNPs.

摘要

已经研究了功能化药物递送系统以改善治疗药物的靶向性和细胞内易位。我们开发了满足独特要求的高功能和高质量脂质，专注于使用微流体装置制备靶向纳米粒子的功能性脂质的质量。在寻找在溶剂中具有高溶解性和分散性的脂质时，这是要求之一，我们注意到 KK-(EK) ₄ -脂质赋予聚乙二醇 (PEG) 修饰（聚乙二醇化）脂质体非特异性细胞缔合，例如细胞穿透肽 (CPP)。我们调查了 KK-(EK) ₄- 脂质具有接近中性的电荷，是一种新型 CPP 修饰的脂质，可增强纳米粒子的细胞内易位。_{然而，KK-(EK) 4} -脂质的细胞结合机制是未知的。因此，我们根据KK-(EK) _{4 -脂质的序列合成了(EK) n} -脂质衍生物，并确定了参与细胞结合的序列位点。此外，将 KK-(EK) ₄ -脂质应用于细胞外囊泡 (EV) 和 mRNA 封装的脂质纳米颗粒 (mRNA-LNP)。与未修饰的相比，KK-(EK) _{4 -脂质修饰的 EV 和 mRNA-LNP 分别显示出更高的细胞结合和体外蛋白表达。}我们阐明了 KK-(EK) ₄-脂质有可能适用于脂质体、EV 和 mRNA-LNP 的细胞内递送。