用于基因转染的自组装聚乙二醇两亲性多肽

P. Klemm, Mira Behnke, Jana I. Solomun, Colin Bonduelle, S. Lecommandoux, Anja Traeger, S. Schubert
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引用次数: 2

摘要

在本研究中,设计了三种由聚乙二醇嵌段和共肽嵌段组成的可生物降解嵌段共聚物,这些嵌段共聚物含有不同组成的阳离子l-赖氨酸(L-Lys)和疏水性苄-l -谷氨酸(Bzl-L-Glu),用于基因传递。采用开环聚合(ROP)法制备多肽,并对Boc-L-Lys侧链进行正交脱保护后,聚合物具有两亲性。为了结合或封装质粒DNA (pDNA),研究了不同的配方:纳米沉淀和乳液技术,使用各种有机溶剂以及水ph控制配方方法。通过动态光散射(DLS)监测复合物和纳米颗粒(NP)的形成,通过凝胶电泳和随后与肝素的控释显示pDNA的相互作用。进一步测试了多肽的细胞毒性和生物降解性。这些复合物和NPs在HEK293T细胞中的转染效率随后被评估为最有希望的大小分布和pDNA结合能力。L-Lys含量最高、疏水螺旋结构(P1*)比例最低的多肽水溶液转染效率最高,因此,该多肽是生物可降解基因传递载体高效基因传递的有希望的候选物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Self-Assembled PEGylated Amphiphilic Polypeptides for Gene Transfection
In the present study, three biodegradable block copolymers composed of a poly(ethylene glycol) block and a copolypeptide block with varying compositions of cationic L-lysine (L-Lys) and hydrophobic benzyl-L-glutamate (Bzl-L-Glu) were designed for gene delivery applications. The polypeptides were synthesized by ring opening polymerization (ROP) and after orthogonal deprotection of Boc-L-Lys side chains, the polymer exhibited an amphiphilic character. To bind or encapsulate plasmid DNA (pDNA), different formulations were investigated: a nanoprecipitation and an emulsion technique using various organic solvents as well as an aqueous pH-controlled formulation method. The complex and nanoparticle (NP) formations were monitored by dynamic light scattering (DLS), and pDNA interaction was shown by gel electrophoresis and subsequent controlled release with heparin. The polypeptides were further tested for their cytotoxicity as well as biodegradability. The complexes and NPs presenting the most promising size distributions and pDNA binding ability were subsequently evaluated for their transfection efficiency in HEK293T cells. The highest transfection efficiencies were obtained with an aqueous formulation of the polypeptide containing the highest L-Lys content and lowest proportion of hydrophobic, helical structures (P1*), which is therefore a promising candidate for efficient gene delivery by biodegradable gene delivery vectors.
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