PH响应型聚丙烯酰吗啉基纳米药物载体的合成与表征
发布时间：2019-01-30 20:07
【摘要】：近年来,聚合物纳米药物载体在癌症治疗领域巨大的应用潜力使其受到广泛关注,肿瘤微环境和正常器官、组织间的差异使环境响应性纳米药物载体备受研究者的青睐,利用其刺激响应特性,可以实现抗肿瘤纳米药物的可控释放和靶向给药,显著提高治癌效率。本文基于生物相容性良好的丙烯酰吗啉(ACMO)和pH敏感的2-(二异丙基氨基)乙基甲基丙烯酸酯(DPA)两种主要单体采用RAFT聚合设计合成了两种新型的pH敏感聚合物纳米药物载体,并对其结构和性能进行了研究,期望能为纳米药物载体的设计和合成提供有利参考。研究内容如下:(1)通过两步RAFT聚合方法,设计合成了一系列结构可控具有不同PDPA链段长度的两亲性嵌段聚合物PACMO-b-PDPA,FT-IR、1H-NMR和GPC测试验证了单体和聚合物的结构;纳米沉淀技术制备的具有不同结构组成的PACMO-b-PDPA NPs的DLS和TEM测试得知,其形态近似球形,并且NPs粒径随着PDPA链段长度增大而变大;以DOX为模型药物,利用UV-Vis对聚合物NPs的载药情况研究发现,实际载药量均在4%左右,载药效率较低;DLS和UV-Vis对PACMO-b-PDPA NPs体外稳定性和蛋白吸附研究发现,其与BSA缓冲液混合44h内的粒径无明显变化,具有良好的抗蛋白吸附能力。(2)以ACMO、DPA、甘氨酸乙酯丙烯酰胺(EGAA)为共聚单体,通过两步RAFT聚合制备两亲性PACMO-b-P(DPA-co-EGAA)(PADE)聚合物。水合肼处理聚合物,使其与DOX上的羰基反应生成pH敏感的腙键,得到双pH敏感 PACMO-b-P(DPA-co-EGAAhyDOX)(PADEhyDox)前药。FT-IR、1H-NMR与GPC测试验证了单体和聚合物的结构;利用UV-Vis测得聚合物前药中DOX的含量分别为10.41%、13.37%和18.44%,与前一章相比这种前药载体提高了DOX药物负载量;DLS和TEM测试显示,聚合物前药在水溶液中自组装成核壳结构的球形NPs,并且粒径小于200 nm,满足纳米药物载体在体内循环的粒径要求;体外模拟药物释放显示,在酸性缓冲液中DOX释放速度、释放量均很大,而在中性缓冲液中只有少量DOX被释放出来,表现出可控释放的性能;通过CLSM和流式细胞仪观察到前药NPs可以快速有效地通过内吞作用运载DOX到细胞核,并在细胞内快速释放DOX使其发挥疗效;MTT法检测表明聚合物无毒,表现出良好的生物相容性,聚合物前药具有很强的抑制细胞增殖的能力。
[Abstract]:In recent years, polymer nano-drug carriers have attracted wide attention due to their great application potential in the field of cancer treatment. The differences between tumor microenvironment and normal organs and tissues make environmental responsive nano-drug carriers attractive to researchers. The controlled release and targeted administration of anti-tumor nanopharmaceuticals can be realized by using their stimulative response, and the efficiency of cancer treatment can be improved significantly. In this paper, based on two main monomers, acrylmorpholine (ACMO) with good biocompatibility and pH sensitive 2- (diisopropylamino) ethyl methacrylate (DPA), two novel pH sensitive monomers were synthesized by RAFT polymerization. Polymer nano-drug carrier, The structure and properties of Nano-drug carriers are studied, which is expected to provide a useful reference for the design and synthesis of nano-drug carriers. The main contents are as follows: (1) A series of amphiphilic block polymers PACMO-b-PDPA,FT-IR, with controllable structure and different length of PDPA segments were designed and synthesized by two-step RAFT polymerization. The structure of monomer and polymer was verified by 1H-NMR and GPC. The DLS and TEM measurements of PACMO-b-PDPA NPs with different structure prepared by nano-precipitation technique showed that the morphology of NPs was approximately spherical, and the diameter of NPs increased with the length of PDPA segment. Using DOX as the model drug and using UV-Vis to study the drug loading of polymer NPs, it was found that the actual loading amount was about 4%, and the drug loading efficiency was low. The in vitro stability and protein adsorption of PACMO-b-PDPA NPs by DLS and UV-Vis showed that the particle size of PACMO-b-PDPA NPs mixed with BSA buffer did not change significantly within 44 h and had good anti-protein adsorption ability. (2) ACMO,DPA,. Amphiphilic PACMO-b-P (DPA-co-EGAA) (PADE) polymer was prepared by two-step RAFT polymerization of glycine ethyl acrylamide (EGAA) as a copolymerization monomer. Hydrazine hydrate was used to react with carbonyl group on DOX to form pH sensitive Hydrazone bond, and then double pH sensitive PACMO-b-P (DPA-co-EGAAhyDOX) (PADEhyDox) prodrug) was obtained. The structure of monomer and polymer was verified by FT-IR,1H-NMR and GPC tests. The contents of DOX in polymer prodrug were 10.41% and 18.44%, respectively, by UV-Vis. Compared with the previous chapter, this prodrug carrier increased the drug load of DOX. The results of DLS and TEM showed that the spherical NPs, with self-assembled core-shell structure of polymer prodrug in aqueous solution and the particle size less than 200 nm, could meet the particle size requirement of nano-drug carrier circulating in vivo. In vitro simulated drug release showed that the release rate and amount of DOX in acidic buffer were very large, but only a small amount of DOX was released in neutral buffer, showing controllable release performance. CLSM and flow cytometry showed that prodrug NPs could transport DOX to the nucleus quickly and effectively through endocytosis and release DOX into cell to make it effective. MTT assay showed that the polymer was nontoxic and showed good biocompatibility. The polymer prodrug had a strong ability to inhibit cell proliferation.
【学位授予单位】：天津工业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TB383.1;TQ460.4
本文编号：2418457
[Abstract]:In recent years, polymer nano-drug carriers have attracted wide attention due to their great application potential in the field of cancer treatment. The differences between tumor microenvironment and normal organs and tissues make environmental responsive nano-drug carriers attractive to researchers. The controlled release and targeted administration of anti-tumor nanopharmaceuticals can be realized by using their stimulative response, and the efficiency of cancer treatment can be improved significantly. In this paper, based on two main monomers, acrylmorpholine (ACMO) with good biocompatibility and pH sensitive 2- (diisopropylamino) ethyl methacrylate (DPA), two novel pH sensitive monomers were synthesized by RAFT polymerization. Polymer nano-drug carrier, The structure and properties of Nano-drug carriers are studied, which is expected to provide a useful reference for the design and synthesis of nano-drug carriers. The main contents are as follows: (1) A series of amphiphilic block polymers PACMO-b-PDPA,FT-IR, with controllable structure and different length of PDPA segments were designed and synthesized by two-step RAFT polymerization. The structure of monomer and polymer was verified by 1H-NMR and GPC. The DLS and TEM measurements of PACMO-b-PDPA NPs with different structure prepared by nano-precipitation technique showed that the morphology of NPs was approximately spherical, and the diameter of NPs increased with the length of PDPA segment. Using DOX as the model drug and using UV-Vis to study the drug loading of polymer NPs, it was found that the actual loading amount was about 4%, and the drug loading efficiency was low. The in vitro stability and protein adsorption of PACMO-b-PDPA NPs by DLS and UV-Vis showed that the particle size of PACMO-b-PDPA NPs mixed with BSA buffer did not change significantly within 44 h and had good anti-protein adsorption ability. (2) ACMO,DPA,. Amphiphilic PACMO-b-P (DPA-co-EGAA) (PADE) polymer was prepared by two-step RAFT polymerization of glycine ethyl acrylamide (EGAA) as a copolymerization monomer. Hydrazine hydrate was used to react with carbonyl group on DOX to form pH sensitive Hydrazone bond, and then double pH sensitive PACMO-b-P (DPA-co-EGAAhyDOX) (PADEhyDox) prodrug) was obtained. The structure of monomer and polymer was verified by FT-IR,1H-NMR and GPC tests. The contents of DOX in polymer prodrug were 10.41% and 18.44%, respectively, by UV-Vis. Compared with the previous chapter, this prodrug carrier increased the drug load of DOX. The results of DLS and TEM showed that the spherical NPs, with self-assembled core-shell structure of polymer prodrug in aqueous solution and the particle size less than 200 nm, could meet the particle size requirement of nano-drug carrier circulating in vivo. In vitro simulated drug release showed that the release rate and amount of DOX in acidic buffer were very large, but only a small amount of DOX was released in neutral buffer, showing controllable release performance. CLSM and flow cytometry showed that prodrug NPs could transport DOX to the nucleus quickly and effectively through endocytosis and release DOX into cell to make it effective. MTT assay showed that the polymer was nontoxic and showed good biocompatibility. The polymer prodrug had a strong ability to inhibit cell proliferation.
【学位授予单位】：天津工业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TB383.1;TQ460.4
【参考文献】
相关期刊论文 前1条
1 平其能;纳米药物和纳米载体系统[J];中国新药杂志;2002年01期
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