Wang Yanfang, Luo Jie, Truebenbach Ines, Reinhard Sören, Klein Philipp Michael, Höhn Miriam, Kern Sarah, Morys Stephan, Loy Dominik M, Wagner Ernst, Zhang Wei
Department of Pharmacy, Ludwig-Maximilians-Universität (LMU) München, Butenandtstrasse 5-13, 81377 Munich, Germany.
Nanosystems Initiative Munich (NIM), Schellingstrasse 4, 80799 Munich, Germany.
ACS Biomater Sci Eng. 2020 Feb 10;6(2):1074-1089. doi: 10.1021/acsbiomaterials.9b01904. Epub 2020 Jan 9.
Sequence-defined lipo-oligomers generated via solid-phase assisted synthesis have been developed as siRNA delivery systems for RNA-interference (RNAi) based gene silencing. Here, novel siRNA lipo-polyplexes were established, which were postmodified with monovalent or bivalent DBCO-PEG agents terminated with peptide GE11 (YHWYGYTPQNVI) for epidermal growth factor receptor (EGFR)-targeted siRNA delivery into EGFR-positive tumor cells. Lipo-oligomers containing eight cationizable succinoyltetraethylene-pentamine (Stp) units mediated higher siRNA nanoparticle core stability than those containing four Stp units, and the incorporation of histidines for enhanced endosomal buffer capacity resulted in an improved gene silencing efficiency. Lipo-polyplexes modified with monovalent or bivalent PEG-GE11 via the copper-free click reaction possessed significantly enhanced cellular internalization and transfection efficiency in EGF receptor-positive human cervical KB and hepatoma Huh7 cells in comparison with the corresponding lipo-polyplexes shielded with PEG without targeting. Furthermore, modification with the bivalent DBCO-PEG-GE11 ligand resulted in higher gene silencing efficiency than modification with the same equivalents of the monovalent DBCO-PEG-GE11 ligand.
通过固相辅助合成产生的序列定义的脂寡聚物已被开发为用于基于RNA干扰（RNAi）的基因沉默的siRNA递送系统。在此，建立了新型的siRNA脂多聚体，其用单价或二价的、以肽GE11（YHWYGYTPQNVI）终止的DBCO-PEG试剂进行后修饰，用于将靶向表皮生长因子受体（EGFR）的siRNA递送至EGFR阳性肿瘤细胞。含有八个可阳离子化的琥珀酰四亚乙基五胺（Stp）单元的脂寡聚物介导的siRNA纳米颗粒核心稳定性高于含有四个Stp单元的脂寡聚物，并且引入组氨酸以增强内体缓冲能力导致基因沉默效率提高。与用无靶向作用的PEG屏蔽的相应脂多聚体相比，通过无铜点击反应用单价或二价PEG-GE11修饰的脂多聚体在EGF受体阳性的人宫颈KB细胞和肝癌Huh7细胞中具有显著增强的细胞内化和转染效率。此外，用二价DBCO-PEG-GE11配体修饰比用相同当量的单价DBCO-PEG-GE11配体修饰导致更高的基因沉默效率。
