基于β-环糊精接枝超支化聚醚的纳米促渗剂对胰岛素局部递送研究

发布时间：2018-07-07 23:31

本文选题：超支化聚缩水甘油醚 + β-环糊精　；参考：《天津医科大学》2017年硕士论文

【摘要】：目的胰岛素频繁皮下注射易引起局部炎症、皮下脂肪萎缩以及脂肪性营养不良等,给患者带来很大痛苦。因此,迫切需要开发胰岛素的非注射给药技术。透皮给药途径因其具有易于吸收、避免肝脏首过效应、实现药物控制释放以及提高患者依从性等优点,已经成为该领域中研究的热点。本文旨在合成一种新型的纳米促渗剂,增加胰岛素的透皮给药,实现血糖的良好控制。方法采取通过阴离子开环聚合的方法合成超支化聚缩水甘油醚(HPG),进一步引入β-环糊精(β-CD),得到HPG-g-CD共聚物。通过纳米技术和自组装方法,构筑出负载胰岛素的纳米粒子,并采用动态光散射对其粒径和电势进行分析;并评价其载药量和包封率,对胰岛素体外释放行为进行研究;通过实验大鼠行活体及离体透皮实验,运用激光共聚焦显微镜来定性观察胰岛素的透皮效果,并通过高效液相色谱仪来定量分析胰岛素透过皮肤的渗透量;采用苏木精-伊红染色法(HE)染色法从组织病理学角度方面测定实验材料对大鼠皮肤的刺激性,从而全面评价HPG-g-CD纳米粒子作为药物透皮递送胰岛素的可行性。结果通过调节HPG-NH2和6-OTs-β-CD质量比,筛选出载药性能较好的HPG-g-CD3聚合物,通过核磁和红外光谱并证实。该聚合物可自组装成纳米粒子,粒径约为200 nm,zeta电势+28.3~+45.5 mV,而且纳米粒子粒径随β-CD比例的增高而增大;该纳米粒可有效地负载胰岛素,包封率可高达85%,且胰岛素包封率随着β-CD的增加逐渐增大,而载药量的差异不显著;胰岛素的体外释放实验结果显示,在pH 7.4条件,随着β-CD的增加,胰岛素释放速率和累积释放百分率均增加;动物透皮实验结果显示,随着时间的延长,FITC标记的胰岛素载药组对活体大鼠皮肤渗透可达真皮层,但游离胰岛素悬液组仅停留在皮肤表层,说明HPG-g-CD促进胰岛素的吸收;体外大鼠透皮实验结果亦显示,负载胰岛素的HPG-g-CD纳米粒子组在相同时间内,胰岛素渗透量显著高于胰岛素溶液组,进一步证实HPG-g-CD纳米粒能够增加胰岛素的透皮吸收。而且该纳米粒不会诱导组织炎症,呈现出良好的生物相容性。结论本文中制备出生物相容性良好的HPG-g-CD共聚物,通过自组装技术构筑出分布均一纳米粒,可有效地负载胰岛素,促进胰岛素透皮吸收,且不诱导组织炎症。该纳米吸收促进剂为胰岛素给药提供新的思路和新的策略,为临床胰岛素安全、有效地治疗糖尿病提新的理论依据。
[Abstract]:Objective the frequent subcutaneous injection of insulin may cause local inflammation, subcutaneous fat atrophy and fatty malnutrition, which bring great pain to the patients. Therefore, there is an urgent need to develop non-injection insulin delivery technology. Because of its advantages of easy absorption, avoiding liver first-pass effect, controlling drug release and improving patient compliance, transdermal drug delivery has become a hot topic in this field. This paper aims to synthesize a new nano osmotic enhancer, to increase the transdermal delivery of insulin, and to control blood sugar well. Methods Hyperbranched polyglycidyl ether (HPG) was synthesized by anion ring-opening polymerization, and 尾 -cyclodextrin (尾 -CD) was further introduced to obtain HPG-g-CD copolymer. The nanoparticles loaded with insulin were constructed by nanotechnology and self-assembly, and their particle size and potential were analyzed by dynamic light scattering (DLS), and the drug loading and encapsulation efficiency were evaluated to study the release behavior of insulin in vitro. The transdermal effect of insulin was qualitatively observed by laser confocal microscope in vivo and in vitro, and the permeation of insulin through the skin was quantitatively analyzed by high performance liquid chromatography (HPLC). In order to evaluate the feasibility of HPG-g-CD nanoparticles as a drug for transdermal delivery of insulin, the irritation of experimental materials to rat skin was determined by hematoxylin-eosin staining (HE) from histopathological point of view. Results by adjusting the mass ratio of HPG-NH2 and 6-OTs- 尾 -CD, HPG-g-CD3 polymers with better drug loading properties were selected. The results were confirmed by NMR and IR spectra. The polymer can self-assemble into nanocrystalline particles, the particle size is about 28.3 ~ 45.5 MV, and the particle size increases with the increase of 尾 -CD ratio. The encapsulation efficiency of insulin increased gradually with the increase of 尾 -CD, but there was no significant difference in drug loading. The results of insulin release in vitro showed that, at pH 7.4, the entrapment rate of insulin increased with 尾 -CD. Both the rate of insulin release and the percentage of cumulative release were increased, and the results of transdermal experiments showed that the FITC labeled insulin loaded group could penetrate into the dermis of living rat skin with the prolongation of time. However, the free insulin suspension group only stayed on the skin surface, indicating that HPG-g-CD promoted the absorption of insulin. The results of rat transdermal experiments in vitro also showed that the HPG-g-CD nanoparticles loaded with insulin at the same time, The amount of insulin permeation was significantly higher than that in insulin solution group, which further confirmed that HPG-g-CD nanoparticles could increase the transdermal absorption of insulin. Moreover, the nanoparticles do not induce tissue inflammation and present good biocompatibility. Conclusion the HPG-g-CD copolymers with good biocompatibility were prepared in this paper. The homogenous nanoparticles were constructed by self-assembly technique, which can effectively load insulin, promote the transdermal absorption of insulin, and do not induce tissue inflammation. The nano-absorption accelerator provides new ideas and strategies for insulin administration and provides a new theoretical basis for clinical insulin safety and effective treatment of diabetes.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.1

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