一种靶向—穿膜肽的设计、合成及活性研究

发布时间：2018-04-28 02:23

本文选题：药物递送系统 + 细胞穿膜肽　；参考：《兰州大学》2017年硕士论文

【摘要】：药物递送系统是指在空间、时间及剂量上全面调控药物在体内分布的技术体系。现在市场上的许多药物,特别是需要跨过细胞膜进入细胞核内发挥作用的抗肿瘤药物,普遍存在的问题是跨膜效率低和药物靶向性差,由此引起的一系列副反应,影响了药物的应用和发展。细胞穿膜肽,是由几个到四十几个氨基酸组成的多肽分子,可以跨过细胞膜进入细胞,自发现以来,以其低毒、高效的跨膜特性受到业内的广泛关注。目前大量研究已经证实,环形细胞穿膜肽具有更好的跨膜特征。细胞靶向肽,利用靶向细胞和非靶向细胞之间的差异,如受体表达类型、pH等,选择性的定位到特定病灶部位,协助药物更好的发挥生物学作用。本课题中我们利用环形细胞穿膜肽与细胞靶向肽相连,以提高其对肿瘤细胞的靶向性和携带药物穿过细胞膜的能力,从而降低对正常细胞的毒副作用。流式细胞实验,药物浓度为5μmol/L,作用时间2h,结果表面多肽载体可以携带更多的荧光(FITC)标记进入细胞,而且相同条件下的激光共聚焦实验显示,酰胺成环cTAT和cR9类似物在DU-145、U251和PC-3肿瘤细胞系中不仅可以进入细胞并在细胞核中有明显分布,这与FITC对照组只是黏附在细胞膜表面形成鲜明的对比,并且可以观察到细胞形态发生明显变化,这是细胞死亡的开始。刃天青实验,药物载体浓度为50μmol/L,25μmol/L和12.5μmol/L,实验结果表明单独的多肽药物载体对非肿瘤细胞HEK-293A细胞具有较低的杀伤作用,细胞的存活率在80%以上,肿瘤细胞系U251细胞存活60%以上。综上所述,多肽载体可以携带更多的荧光进入细胞并且在细胞核中有大量分布,可以促进肿瘤细胞的死亡,这种由内而外的肿瘤细胞的内源性凋亡,促进多肽药物载体在药物递送系统中的应用。此外,多肽与抗肿瘤药物阿霉素腙键连接,进一步为多肽载体在成药方面的研究提供参考价值。
[Abstract]:Drug delivery system (DDS) is a technical system that regulates the distribution of drugs in vivo in space, time and dose. Many drugs on the market today, especially anti-tumor drugs that need to cross the cell membrane to work in the nucleus, have the common problem of low transmembrane efficiency and poor drug targeting, resulting in a series of side effects. It affects the application and development of drugs. Cellular transmembrane peptides, composed of several to forty amino acids, can cross cell membranes and enter cells. Since their discovery, they have attracted wide attention for their low toxic and efficient transmembrane properties. At present, a large number of studies have confirmed that annular cell transmembrane peptide has better transmembrane characteristics. Cell-targeted peptides, using the difference between target cells and non-target cells, such as receptor expression type, pH and so on, selectively locate the specific focus to assist the drug to play a better biological role. In order to improve the ability of targeting tumor cells and carry drugs through the cell membrane, we use circular cell membrane peptide to connect with cell targeted peptide, so as to reduce the toxic and side effects on normal cells. Flow cytometry showed that the drug concentration was 5 渭 mol / L and the time was 2 h. The results showed that the surface polypeptide carrier could carry more fluorescent FITC- labeled cells into the cells, and the confocal laser experiments under the same conditions showed that, The cyclic cTAT and cR9 analogues were not only found in DU-145U251 and PC-3 tumor cell lines, but also distributed in the nucleus, which was significantly different from that of the FITC control group, which only adhered to the surface of the cell membrane. A marked change in cell morphology was observed, which was the beginning of cell death. The concentration of drug carrier was 50 渭 mol / L ~ 25 渭 mol/L and 12.5 渭 mol / L respectively. The results showed that the single polypeptide drug carrier had a low killing effect on non-tumor HEK-293A cells, the survival rate of the cells was over 80%, and the survival rate of tumor cell line U251 was more than 60%. To sum up, polypeptide vectors can carry more fluorescence into cells and have a large distribution in the nucleus, which can promote the death of tumor cells, the endogenous apoptosis of tumor cells from inside to outside. To promote the application of polypeptide drug carriers in drug delivery systems. In addition, polypeptides are linked to adriamycin Hydrazone, which provides a reference for the study of polypeptide carriers in proprietary medicine.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R91;R96

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