D型IC-8多肽修饰的还原敏感型RHB自组装双靶向核酸递送载体的研究

针对非病毒基因递送载体的靶向性、稳定性、安全性和高效性等关键问题，设计并构建高效、稳定、低毒的双靶向三元复合物非病毒载体。本课题拟采用还原敏感型超树枝化聚胺基-胺 (RHB) 作为载体材料与质粒DNA或siRNA自组装压缩成纳米级复合物，以靶向肿瘤血管上皮细胞Annexins受体的D型IC-8肽修饰的透明质酸(HA)对该复合物进行静电包衣，形成靶向肿瘤血管和肿瘤细胞CD44的双靶向还原敏感性IC-8-PEG-HA/RHB/DNA或siRNA三元复合物系统。采用现代药剂学技术对该系统的制备方法进行筛选和优化；利用小角度X衍射对三元复合物的二维结构进行表征，阐明该系统进入细胞的途径和机制；采用激光共聚焦技术和活体成像技术考察该系统在细胞内的转运情况和肿瘤靶向性。从细胞水平和整体动物水平评价该载体系统的转染效率和靶向治疗肿瘤的应用效果。

To increase the targeting, stability, security and efficiency of non-viral gene delivery vector, we design an efficient, stable, low toxicity and dual-targeting ternary complex nanoparticles as non-viral gene delivery system. Our interest has been focused on a redox-sensitive dendritic poly amino-amine (RHB) as carrier material which condense DNA or siRNA self-assembly into nano-complexes. We hypothesized D-IC-8 peptide which target to the Annexins receptor of tumor vascular endothelial cells modified hyaluronic acid (HA) coated RHB/DNA or siRNA complexes to form CD44 targeting and tumor vasculature targeting dual targeting core-shell redox-sensitive IC-8-PEG-HA/RHB/DNA or siRNA ternary complexes system based on electrostatic self-assembly adsorbed coatings. The screening and optimization of the system formulation were performed with modern pharmacy technology; the complexes were characterized by small-angle X-ray diffraction on two-dimensional structure; the ways and mechanisms of the cellular uptake would be clarified; we will use confocal microscopy to examine the intracellular fate and targeting. The transfection efficiency and targeting of the system will be evaluated on the cellular and whole animal level.