Lab of Biocompatible Polymers, Department of Biological, Chemical and Pharmaceutical Sciences and Technologies (STEBICEF), University of Palermo, Via Archirafi 32, 90123 Palermo, Italy.
Department of Pharmacy, University of Salerno, Via Giovanni Paolo II, 132, 84084 Fisciano, Italy.
Molecules. 2021 Mar 19;26(6):1713. doi: 10.3390/molecules26061713.
Here, a novel strategy of formulating efficient polymeric carriers based on the already described INU-IMI-DETA for gene material whose structural, functional, and biological properties can be modulated and improved was successfully investigated. In particular, two novel derivatives of INU-IMI-DETA graft copolymer were synthesized by chemical functionalisation with epidermal growth factor (EGF) or polyethylenglycol (PEG), named INU-IMI-DETA-EGF and INU-IMI-DETA-PEG, respectively, in order to improve the performance of already described "inulin complex nanoaggregates" (ICONs). The latter were thus prepared by appropriately mixing the two copolymers, by varying each component from 0 to 100 wt% on the total mixture, named EP-ICONs. It was seen that the ability of the INU-IMI-DETA-EGF/INU-IMI-DETA-PEG polymeric mixture to complex siGL3 increases with the increase in the EGF-based component in the EP-ICONs and, for each sample, with the increase in the copolymer:siRNA weight ratio (R). On the other hand, the susceptibility of loaded siRNA towards RNase decreases with the increase in the pegylated component in the polymeric mixture. At all R values, the average size and the zeta potential values are suitable for escaping from the RES system and suitable for prolonged intravenous circulation. By means of biological characterisation, it was shown that MCF-7 cells are able to internalize mainly the siRNA-loaded into EGF-decorated complexes, with a significant difference from ICONs, confirming its targeting function. The targeting effect of EGF on EP-ICONs was further demonstrated by a competitive cell uptake study, i.e., after cell pre-treatment with EGF. Finally, it was shown that the complexes containing both EGF and PEG are capable of promoting the internalisation and therefore the transfection of siSUR, a siRNA acting against surviving mRNA, and to increase the sensitivity to an anticancer agent, such as doxorubicin.
在这里，成功研究了一种基于已描述的 INU-IMI-DETA 的高效聚合物载体的新型策略，用于基因材料，其结构、功能和生物学性质可以被调节和改善。特别是，通过化学官能化用表皮生长因子（EGF）或聚乙二醇（PEG）合成了两种新型 INU-IMI-DETA 接枝共聚物的衍生物，分别命名为 INU-IMI-DETA-EGF 和 INU-IMI-DETA-PEG，以提高已描述的“菊粉复合纳米聚集体”（ICONs）的性能。后者通过适当混合两种共聚物来制备，通过在总混合物中从 0 到 100wt%变化每个成分，命名为 EP-ICONs。结果表明，INU-IMI-DETA-EGF/INU-IMI-DETA-PEG 聚合物混合物与 siGL3 复合的能力随 EP-ICONs 中基于 EGF 的成分的增加而增加，并且对于每个样品，随共聚物：siRNA 重量比（R）的增加而增加。另一方面，负载的 siRNA 对 RNase 的敏感性随聚合物混合物中 PEG 化成分的增加而降低。在所有 R 值下，平均粒径和 zeta 电位值适合逃避 RES 系统并适合延长静脉内循环。通过生物学特性研究表明，MCF-7 细胞能够主要内化负载到 EGF 修饰的复合物中的 siRNA，与 ICONs 有显著差异，证实了其靶向功能。通过竞争细胞摄取研究进一步证明了 EGF 对 EP-ICONs 的靶向作用，即细胞用 EGF 预处理后。最后，结果表明，同时包含 EGF 和 PEG 的复合物能够促进 siSUR 的内化和转染，siSUR 是一种针对存活 mRNA 的 siRNA，并增加对阿霉素等抗癌剂的敏感性。
