Département de Chimie, Université de Montréal, C.P. 6128, Succ. Centre-ville, Montréal, QC H3C 3J7, Canada; Faculté de Pharmacie, Université de Montréal, C.P. 6128, Succ. Centre-ville, Montréal, QC H3C 3J7, Canada.
Faculté de Pharmacie, Université de Montréal, C.P. 6128, Succ. Centre-ville, Montréal, QC H3C 3J7, Canada; Gene Delivery Laboratory, Faculté de pharmacie, Université de Montréal, H3C 3J7 Montréal, QC, Canada.
Int J Pharm. 2020 Mar 30;578:119078. doi: 10.1016/j.ijpharm.2020.119078. Epub 2020 Jan 24.
Gene therapy is a promising tool for the treatment of various cancers but is hindered by the physico-chemical properties of siRNA and needs a suitable vector for the delivery of siRNA to the target tissue. Bile acid-based block copolymers offers certain advantages for the loading and delivery of siRNA since they can efficiently complex siRNA and bile acids are biocompatible endogenous molecules. In this study, we demonstrate the use of lipids as co-surfactants for the preparation of mixed micelles to improve the siRNA delivery of cholic acid-based block copolymers. Poly(allyl glycidyl ether) (PAGE) and poly(ethylene glycol) (PEG) were polymerized on the surface of cholic acid to afford a star-shaped block copolymer with four arms (CA-PAGE-b-PEG). The allyl groups of PAGE were functionalized to bear primary or tertiary amines and folic acid was grafted onto the PEG chain end to increase cell uptake. (CA-PAGE-b-PEG) functionalized with either primary or tertiary amines show high siRNA complexation with close to 100% complexation at N/P ratio of 8. Uniform aggregates with diameters between 181 and 188 nm were obtained. DOPE, DSPE-PEG, and DSPE-PEG lipids were added as co-surfactants to help stabilize the nanoparticles in the cell culture media. Mixed micelles had high siRNA loading with close to 100% functionalization at N/P ratio of 16 and diameters ranging from 153 to 221 nm. The presence of lipids in the mixed micelles improved cell uptake with a concomitant siRNA transfection in HeLa and HeLa-GFP model cells, respectively.
基因治疗是治疗各种癌症的一种很有前途的工具，但受到 siRNA 的物理化学性质的限制，需要合适的载体将 siRNA 递送到靶组织。基于胆汁酸的嵌段共聚物在负载和递送 siRNA 方面具有一定的优势，因为它们可以有效地将 siRNA 复合，并且胆汁酸是生物相容的内源性分子。在这项研究中，我们展示了使用脂质作为共乳化剂来制备混合胶束，以提高基于胆酸的嵌段共聚物的 siRNA 递呈效率。聚（烯丙基缩水甘油醚）（PAGE）和聚乙二醇（PEG）接枝在胆酸表面上，得到具有四个臂的星形嵌段共聚物（CA-PAGE-b-PEG）。PAGE 的烯丙基基团被功能化，以承载伯胺或叔胺，并且叶酸被接枝到 PEG 链末端，以增加细胞摄取。具有伯胺或叔胺的（CA-PAGE-b-PEG）表现出与 siRNA 的高复合率，在 N/P 比为 8 时接近 100%的复合率。在 181 到 188nm 之间的直径范围内得到均匀的聚集物。添加 DOPE、DSPE-PEG 和 DSPE-PEG 脂质作为共乳化剂，以帮助在细胞培养介质中稳定纳米颗粒。混合胶束具有高 siRNA 负载，在 N/P 比为 16 时接近 100%的功能化，直径范围从 153 到 221nm。混合胶束中脂质的存在提高了细胞摄取效率，同时在 HeLa 和 HeLa-GFP 模型细胞中分别进行了 siRNA 转染。
