Trehalose click polymers inhibit nanoparticle aggregation and promote pDNA delivery in serum
Trehalose click polymers inhibit nanoparticle aggregation and promote pDNA delivery in serum
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DOI：
10.1021/ja0585580
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发表时间：
2006-06-28
影响因子：
15
通讯作者：
Reineke, Theresa M.
中科院分区：
文献类型：
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作者：
Srinivasachari, Sathya;Liu, Yemin;Reineke, Theresa M.
在此，通过“点击聚合”合成了三种新的糖聚合物，以在含有血清的生物介质存在下促进核酸递送。这些结构被设计为含有海藻糖部分以促进生物相容性、水溶性和抗聚集稳定性，酰胺-三唑基团以增强DNA结合亲和力，以及寡胺单元以促进DNA封装、磷酸盐中和以及与细胞表面的相互作用。 2,3,4,2', 3', 4'-六-O-乙酰基-6,6'-二叠氮基-6,6'-二脱氧-(D)-海藻糖 (4) 单体通过铜 (I) 催化的叠氮化物-炔环加成反应与一系列含有一个、两个或三个 Boc 保护的仲胺的二炔酰胺共聚单体聚合 (7a,分别为 7b 或 7c)。脱保护后，获得了具有相似聚合度 (n = 56-61) 的三种水溶性聚阳离子（9a、9b 或 9c），以阐明胺数对核酸结合、复合物形成、稳定性和细胞递送的作用。凝胶电泳和溴化乙锭实验表明，9a-9c 与质粒 DNA (pDNA) 结合，并以取决于胺数的 N/P 比率形成复合物（聚合复合物）。 TEM 实验表明，9a-9c 复合物很小（50-120 nm），并且具有与聚合物链刚度相关的形态（球形和棒状）。在含有血清的介质存在下的动态光散射研究表明，9c 复合物具有低程度的絮凝，而 9a 和 9b 复合物快速聚集。进一步的生物学研究表明，这些结构具有生物相容性，可以将 pDNA 输送到 HeLa 细胞中。特别是，9c 复合物在血清存在下促进了高递送效率和基因表达谱。
Herein, three new glycopolymers have been synthesized via "click polymerization" to promote nucleic acid delivery in the presence of biological media containing serum. These structures were designed to contain a trehalose moiety to promote biocompatibility, water solubility, and stability against aggregation, amide-triazole groups to enhance DNA binding affinity, and an oligoamine unit to facilitate DNA encapsulation, phosphate neutralization, and interactions with cell surfaces. A 2,3,4,2', 3', 4'-hexa-O-acetyl-6,6'-diazido-6,6'-dideoxy-(D)-trehalose (4) monomer was polymerized via copper(I)-catalyzed azide-alkyne cycloaddition with a series of dialkyne-amide comonomers that contain either one, two, or three Boc-protected secondary amines (7a, 7b, or 7c, respectively). After deprotection, three water-soluble polycations (9a, 9b, or 9c) were obtained with similar degrees of polymerization (n = 56-61) to elucidate the role of amine number on nucleic acid binding, complex formation, stability, and cellular delivery. Gel electrophoresis and ethidium bromide experiments showed that 9a-9c associated with plasmid DNA (pDNA) and formed complexes (polyplexes) at N/P ratios dependent on the amine number. TEM experiments revealed that 9a-9c polyplexes were small (50-120 nm) and had morphologies (spherical and rodlike) associated with the polymer chain stiffness. Dynamic light scattering studies in the presence of media containing serum demonstrated that 9c polyplexes had a low degree of flocculation, whereas 9a and 9b polyplexesd aggregate rapidly. Further biological studies revealed that these structures were biocompatible and deliver pDNA into HeLa cells. Particularly, 9c polyplexes promoted high delivery efficacy and gene expression profiles in the presence of serum.