苯硼酸修饰的聚合物载体的制备及在口服递送胰岛素中的应用研究

发布时间：2018-06-20 11:50

本文选题：口服 + 胰岛素　；参考：《浙江理工大学》2017年硕士论文

【摘要】：近年来,糖尿病成为了全球范围内的一种疾病,它会对人体健康系统造成巨大的损害和沉重的负担。据统计,2015年全球成年人中有4.15亿患有糖尿病,按照这种趋势,到2040年预计将达到6.42亿,这意味着每十个人里面就有一个患有糖尿病。聚合物基纳米药物载体用于药物负载及其可控释放的应用研究受到人们的密切关注。此外,通过目标聚合物的口服给药,不仅可以增强智能响应,还可以减少注射的痛苦,同时具有高效、持续缓释的优点。苯硼酸及其衍生物能与糖类相互作用,形成共价复合物。因此,引入苯硼酸修饰到聚合物药物载体,研究口服利用后的胰岛素释放,具有重要的研究意义。本论文采用三种不同的方法制备了含苯硼酸修饰葡萄糖敏感口服胰岛素载体,并对其药物释放性能进行了研究。具体来说,主要包括以下三个部分:1.结合RAFT聚合与酶法酯交换方法制备苯硼酸修饰葡萄糖敏感的荧光共聚物。这种两亲性的共聚物胶束可以自组装成球形胶束,通过核磁(1H NMR)、红外(FT-IR)、透射电镜(TEM)和动态光散射(DLS)对其进行表征。使用荧光光谱仪和激光共聚焦显微镜(CLSM)对其荧光特性进行了研究。由于其两亲性和葡萄糖敏感的特性,作为胰岛素载体用以包覆胰岛素。在体外模拟葡萄糖进行了胰岛素释放研究。此外,该药物载体具有较低的细胞毒性和较好的抗蛋白稳定性。释放前后的胰岛素经圆二色谱(CD)对比分析,表明释放的胰岛素的二级结构没有损伤。这种苯硼酸修饰葡萄糖敏感的荧光聚合物胶束在胰岛素传递治疗糖尿病方面有望成为有效的载体材料。2.近些年,口服递送胰岛素的研究已经引起人们极大的关注。本章采用低温乳化聚合的方法,首先设计制备了苯硼酸修饰葡萄糖敏感聚合物纳米颗粒,并研究了其对Caco-2细胞的细胞毒性及抗蛋白稳定性,胰岛素在体外模拟pH和葡萄糖刺激释放也进行了研究。为了增强口服胰岛素的利用率,采用透明质酸的交联构建3D水凝胶体系,将负载胰岛素的聚合物纳米颗粒包封其中,以克服吸收屏障和减少运送传递过程中的胰岛素的损失。通过糖尿病小鼠口服给药研究,发现对该种水凝胶体系具有长时间的口服降血糖效果和更高的药物利用率。3.为了克服口服递送胰岛素的吸收屏障,采用羧化壳聚糖接枝聚合的方法,将肠上皮细胞特异性吸收的L-缬氨酸加入体系,制备具有靶向作用的苯硼酸修饰葡萄糖敏感的聚合物。研究发现对HT-29细胞有较低的毒性和较好的抗蛋白稳定性。胰岛素在体外模拟pH和葡萄糖刺激释放也进行了研究。在包含胃蛋白酶的模拟胃液中和包含胰蛋白酶的模拟肠液中对胰岛素的化学稳定性进行了研究。通过激光共聚焦显微镜观察了HT-29细胞对聚合物纳米颗粒的转运行为。进一步通过糖尿病小鼠口服给药研究了作为胰岛素运送载体的优势,表明该L-缬氨酸修饰的基于壳聚糖的纳米颗粒在口服胰岛素的应用中有着巨大的潜力。
[Abstract]:In recent years, diabetes has become a global disease, which can cause huge damage and heavy burden on human health systems. According to statistics, 415 million of the world's adults in 2015 were suffering from diabetes, which is expected to reach 642 million by 2040, which means that one in every ten people is suffering from diabetes. The application of polymer based nanoscale drug carriers to drug loading and controlled release is closely concerned. In addition, oral administration of the target polymer can not only enhance the intelligent response, but also reduce the pain of the injection, at the same time, it has the advantage of high efficiency and continuous slow release. Therefore, the introduction of boric acid to polymer drug carriers and the study of the release of insulin after oral use is of great significance. In this paper, three different methods were used to prepare the oral glucose sensitive oral insulin carrier containing benzyl boric acid, and the drug release performance was studied. It mainly includes the following three parts: 1. the preparation of benzboric acid modified glucose sensitive fluorescent copolymers with RAFT polymerization and enzymatic transesterification. The two Pro copolymer micelles can be self assembled into spherical micelles and are characterized by NMR (1H NMR), infrared (FT-IR), TEM and DLS. Fluorescence spectrometers and laser confocal microscopy (CLSM) have studied their fluorescence characteristics. Due to their two affinity and glucose sensitive properties, they are used as insulin carriers to encapsulate insulin. The insulin release of glucose in vitro is studied in vitro. In addition, the drug carrier has lower cytotoxicity and better anti protein stability. A comparative analysis of the insulin before and after the release by circular two chromatography (CD) shows that the two grade structure of the released insulin is not damaged. This kind of benzyl boric acid modified glucose sensitive fluorescent polymer micelles is expected to become an effective carrier material for insulin delivery in the treatment of diabetes. In recent years, the study of oral delivery of insulin has been cited. In this chapter, we designed and prepared the phallic acid modified glucose sensitive polymer nanoparticles in this chapter, and studied the cytotoxicity and anti protein stability of Caco-2 cells. In order to enhance the oral islet, the insulin in vitro simulated pH and glucose stimulation release. The 3D hydrogel system was constructed by cross-linking of hyaluronic acid. The polymer nanoparticles loaded with insulin were encapsulated to overcome the absorption barrier and reduce the loss of insulin during delivery. Oral administration of diabetic mice showed that the hydrogel system had a long time of oral hypoglycemia. The effect and higher drug utilization rate.3., in order to overcome the absorption barrier of oral insulin delivery, uses carboxyl Chitosan Graft polymerization, adding L- valine, which is specifically absorbed by intestinal epithelial cells, and preparing the polymer sensitive to glucose with benzyl boric acid, which has a targeting effect. The study found that there is a lower toxicity to HT-29 cells. Sex and good anti protein stability. Insulin stimulated release of pH and glucose in vitro was also studied. The chemical stability of insulin was studied in simulated gastric juice containing pepsin neutralization and in simulated intestinal fluid containing trypsin. The effect of HT-29 cells on polymer nanoparticles was observed by laser confocal microscopy. The transport behavior of the particles was further studied by oral administration of the diabetic mice as an insulin carrier, indicating that the L- valine modified nanoparticles based on chitosan have great potential in the application of oral insulin.
【学位授予单位】：浙江理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TQ467.32

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