大黄素壳聚糖硬脂酸纳米胶束的制备及评价

发布时间：2018-01-21 01:03

本文关键词： 壳聚糖硬脂酸大黄素纳米胶束抗肿瘤　出处：《华东理工大学》2017年硕士论文　论文类型：学位论文

【摘要】：在新型药物制剂中,聚合物胶束是研究比较热门的一种剂型。本文利用自主研发的壳聚糖硬脂酸聚合物,制备载大黄素(emodin, EMO)纳米胶束。达到改善EMO水溶性和提高EMO抗肿瘤活性的目的。采用壳聚糖酶将大分子壳聚糖(450 KDa)降解为小分子壳聚糖(18KDa)。以碳二亚胺为交联耦合剂,用疏水性硬脂酸修饰小分子壳聚糖,得到壳寡糖硬脂酸嫁接物(CSO-SA),进而制备载大黄素的纳米胶束。通过不同实验方法评价CSO-SA的氨基取代度(SD%)、临界胶束浓度(CMC)、粒径、Zeta电位。得知CSO-SA聚合物是一种分散性好、粒径小的纳米级胶束。采用FITC对CSO-SA胶束荧光标记,评价胃癌细胞对纳米胶束的经时摄取情况。选用裸鼠为动物模型以DiR为示踪剂,小动物活体成像仪中观察CSO-SA胶束在体内的经时分布。CSO-SA在12h内能被胃癌细胞充分摄取,在体内能有效富集在肿瘤部位。透析法制备CSO-SA/EMO载药胶束,得到电位略有下降,粒径略有上升的载药胶束。CSO-SA能够有效包载EMO,包封率高达96.0%,载药量可达21.2%。高效液相色谱法(HPLC)检测不同时间点CSO-SA/EMO的累积释放量可知,72小时内CSO-SA/EMO胶束累积释放量达78.4%,释放速度和EMO比更为温和、平缓。MTT法和流式细胞术(FCM)证明CSO-SA胶束是一种安全性高的药物载体,CSO-SA/EMO对胃癌细胞凋亡作用明显高于游离EMO。综上所述,自主研发的聚合物胶束生物相容性好、毒性低。能高效包载EMO,显著提高EMO的抗肿瘤活性,是一种具有广阔前景的新型制剂。
[Abstract]:Polymer micelles are one of the most popular formulations in the new drug preparation. In this paper, emodin loaded with emodin was prepared by using the self-developed chitosan stearic acid polymer. In order to improve the water solubility of EMO and enhance the anti-tumor activity of EMO, the macromolecular chitosan (450KDa) was decomposed into small molecular chitosan by using chitosanase. Carbon diimide was used as crosslinking coupling agent. The small molecular chitosan was modified with hydrophobic stearic acid, and the chitosan oligosaccharide stearic acid graft CSO-SAwas obtained. Then the emodin-loaded nano-micelles were prepared. The amino substitution degree of CSO-SA was evaluated by different experimental methods, and the critical micelle concentration was determined as well as its particle size. Zeta potential. It was found that CSO-SA polymer was a kind of nano-micelle with good dispersion and small particle size. FITC was used to label CSO-SA micelles. To evaluate the uptake of nano-micelles by gastric cancer cells, nude mice were used as animal models and DiR was used as tracer. The time distribution of CSO-SA micelles in vivo. CSO-SA was fully ingested by gastric cancer cells within 12 hours. CSO-SA/EMO micelles were prepared by dialysis, and the potential decreased slightly. The drug loaded micelle. CSO-SA with slightly increasing particle size can effectively encapsulate EMO.The encapsulation efficiency is as high as 96.0%. High performance liquid chromatography (HPLC) was used to detect the cumulative release of CSO-SA/EMO at different time points. The cumulative release of CSO-SA/EMO micelles reached 78.4% within 72 hours, and the release rate and EMO release rate were milder than those of EMO. CSO-SA micelles were proved to be a high safety drug carrier by flattening. MTT assay and flow cytometry (FCM). The effect of CSO-SA/EMO on apoptosis of gastric cancer cells was significantly higher than that of free EMO.In conclusion, the self-developed polymer micelles had good biocompatibility, low toxicity and high efficiency in encapsulating EMO. The anti-tumor activity of EMO is improved significantly, and it is a new preparation with broad prospect.
【学位授予单位】：华东理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R943

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