PH響應型聚丙烯酰嗎啉基納米藥物載體的合成與表征
發(fā)布時間：2019-01-30 20:07
【摘要】：近年來,聚合物納米藥物載體在癌癥治療領域巨大的應用潛力使其受到廣泛關注,腫瘤微環(huán)境和正常器官、組織間的差異使環(huán)境響應性納米藥物載體備受研究者的青睞,利用其刺激響應特性,可以實現(xiàn)抗腫瘤納米藥物的可控釋放和靶向給藥,顯著提高治癌效率。本文基于生物相容性良好的丙烯酰嗎啉(ACMO)和pH敏感的2-(二異丙基氨基)乙基甲基丙烯酸酯(DPA)兩種主要單體采用RAFT聚合設計合成了兩種新型的pH敏感聚合物納米藥物載體,并對其結構和性能進行了研究,期望能為納米藥物載體的設計和合成提供有利參考。研究內(nèi)容如下:(1)通過兩步RAFT聚合方法,設計合成了一系列結構可控具有不同PDPA鏈段長度的兩親性嵌段聚合物PACMO-b-PDPA,FT-IR、1H-NMR和GPC測試驗證了單體和聚合物的結構;納米沉淀技術制備的具有不同結構組成的PACMO-b-PDPA NPs的DLS和TEM測試得知,其形態(tài)近似球形,并且NPs粒徑隨著PDPA鏈段長度增大而變大;以DOX為模型藥物,利用UV-Vis對聚合物NPs的載藥情況研究發(fā)現(xiàn),實際載藥量均在4%左右,載藥效率較低;DLS和UV-Vis對PACMO-b-PDPA NPs體外穩(wěn)定性和蛋白吸附研究發(fā)現(xiàn),其與BSA緩沖液混合44h內(nèi)的粒徑無明顯變化,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)為共聚單體,通過兩步RAFT聚合制備兩親性PACMO-b-P(DPA-co-EGAA)(PADE)聚合物。水合肼處理聚合物,使其與DOX上的羰基反應生成pH敏感的腙鍵,得到雙pH敏感 PACMO-b-P(DPA-co-EGAAhyDOX)(PADEhyDox)前藥。FT-IR、1H-NMR與GPC測試驗證了單體和聚合物的結構;利用UV-Vis測得聚合物前藥中DOX的含量分別為10.41%、13.37%和18.44%,與前一章相比這種前藥載體提高了DOX藥物負載量;DLS和TEM測試顯示,聚合物前藥在水溶液中自組裝成核殼結構的球形NPs,并且粒徑小于200 nm,滿足納米藥物載體在體內(nèi)循環(huán)的粒徑要求;體外模擬藥物釋放顯示,在酸性緩沖液中DOX釋放速度、釋放量均很大,而在中性緩沖液中只有少量DOX被釋放出來,表現(xiàn)出可控釋放的性能;通過CLSM和流式細胞儀觀察到前藥NPs可以快速有效地通過內(nèi)吞作用運載DOX到細胞核,并在細胞內(nèi)快速釋放DOX使其發(fā)揮療效;MTT法檢測表明聚合物無毒,表現(xiàn)出良好的生物相容性,聚合物前藥具有很強的抑制細胞增殖的能力。
[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.
【學位授予單位】：天津工業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：TB383.1;TQ460.4
本文編號：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.
【學位授予單位】：天津工業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：TB383.1;TQ460.4
【參考文獻】
相關期刊論文 前1條
1 平其能;納米藥物和納米載體系統(tǒng)[J];中國新藥雜志;2002年01期
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