本文關鍵詞： 納米醫(yī)學 磷酸膽堿兩性離子 聚合物膠束 pH響應 診療一體化　出處：《浙江大學》2016年博士論文　論文類型：學位論文

【摘要】：聚合物納米膠束,由于可以改善藥物溶解度、增加在腫瘤部位靶向富集并提高生物利用度,已成為提高癌癥診斷與治療效果并減少毒副作用的重要納米載體之一。本文依據(jù)磷酸膽堿兩性離子的細胞膜仿生特性,致力于設計合成磷酸膽堿基載體材料,構建以兩性離子磷酸膽堿為親水殼層的聚合物納米膠束,并通過對膠束內核的不同設計實現(xiàn)其在癌癥診斷與治療上的多功能化作用,主體包含以下四個方面:1.研究采用RAFT聚合制備了磷酸膽堿基嵌段共聚物PMPC-b-PDDMMA。該共聚物能自組裝形成單分散的納米膠束,并能在蛋白質和培養(yǎng)基溶液中保持粒徑穩(wěn)定。該共聚物疏水嵌段中的環(huán)狀縮酮基團,可在酸性條件下實現(xiàn)斷裂。體外藥物釋放結果顯示,相比于正常生理環(huán)境,酸性環(huán)境下阿霉素的釋放速度顯著增強。細胞實驗也表明該載藥膠束能有效實現(xiàn)細胞內的阿霉素釋放并抑制HepG2肝癌細胞的增殖。同時,包載IR-780近紅外染料膠束的活體成像結果顯示,磷酸膽堿化膠束能有效富集于腫瘤組織。2.研究進一步基于非共價鍵的主客體作用,設計合成了酸性敏感的磷酸膽堿化聚合物前藥膠束。通過RAFT聚合制備了含有金剛烷的無規(guī)共聚物poly(MPC-co-Ada),并用2DNOESYNMR證實了金剛烷與阿霉素修飾的β-環(huán)糊精(DOX-hyd-CD)之間的主客體絡合作用。體外藥物釋放結果表明,所制備的前藥膠束可在模擬溶酶體/內涵體的酸性條件下通過腙鍵的斷裂實現(xiàn)阿霉素的快速釋放。細胞實驗結果證實了前藥膠束的有效細胞內吞以及對HepG2肝癌細胞增殖的抑制作用。得益于“即插即用”式的主客體作用,該前藥膠束還有望進一步實現(xiàn)多功能化的腫瘤治療效果。3.研究進一步通過RAFT聚合制備了含苯甲醛基團的磷酸膽堿基共聚物,并通過腙鍵接枝了 AIE分子四苯基乙烯(TPE)。所得到的共聚物PMPC-hyd-TPE能自組裝形成以磷酸膽堿為殼、以TPE為核的納米膠束。疏水的TPE分子聚集成核,不但可以激發(fā)AIE效應還能通過疏水相互作用包載阿霉素。體外藥物釋放實驗表明,在模擬細胞內酸性環(huán)境下,腙鍵鍵接TPE的斷裂使得膠束溶脹并快速釋放出阿霉素。細胞實驗顯示,該聚合物膠束具備良好的AIE成像效果并可有效抑制HepG2肝癌細胞的增殖。同時,DLS結果表明磷酸膽堿化膠束在含F(xiàn)BS的DMEM培養(yǎng)基里能保持長效穩(wěn)定,且與血漿蛋白BSA和FBG共孵育24h后,膠束吸附蛋白質的含量維持在較低水平。進一步動物實驗顯示,磷酸膽堿化載藥膠束能靶向富集并傳遞阿霉素至腫瘤組織。4.研究進一步探索了通過簡便的一步聚合以及單一近紅外染料負載制備診療一體化的磷酸膽堿均聚物膠束的可行性。通過增長磷酸膽堿單體MPC的烷基鏈長度得到了兩親性單體MDPC,經一步RAFT聚合便可得到兩親性均聚物PMDPC。該兩親性均聚物PMDPC同樣能自組裝形成穩(wěn)定的膠束,并實現(xiàn)對近紅外染料IR-780的有效負載。經均聚物膠束負載后,近紅外染料IR-780的光穩(wěn)定性顯著增強,并具備高效的光熱轉換效果,能有效殺傷BxPC-3胰腺癌細胞。進一步的動物實驗表明,IR-780包載的磷酸膽堿化均聚物膠束能有效富集于腫瘤區(qū)域,具有高分辨且長效的近紅外成像效果。
[Abstract]:The polymer micelles, which can improve drug solubility and bioavailability to increase and enriched in tumor target, improve the diagnosis and treatment of cancer has become one of the important carrier of nano effect and reduce toxic side effects. Based on the characteristics of cell membrane biomimetic phosphorylcholine zwitterionic, committed to the synthesis of choline phosphate based carrier material design, to construct zwitterionic phosphorylcholine polymer micelles as hydrophilic shell, and through the different design of the micelle core realization of multiple functions in cancer diagnosis and treatment of the main effect, including the following four aspects: 1. study by RAFT polymerization preparation of phosphorylcholine based block copolymers PMPC-b-PDDMMA. the copolymer can self assemble to form micelles dispersed, and can keep the culture medium solution in stable particle size. The copolymer protein and hydrophobic block in cyclic ketal Groups can realize fracture under acidic conditions. The in vitro drug release results showed that, compared to the normal physiological environment, the release rate of adriamycin in acidic environment was significantly enhanced. Cell experiments showed that the micelle can effectively achieve the intracellular release of doxorubicin and inhibit HepG2 liver cancer cell proliferation. At the same time, the results of in vivo imaging encapsulation IR-780 near infrared dye micelles showed that the micelles could effectively enrich bile alkaline phosphate on.2. tumor tissue further based on object function of non covalent bonds, bile acid sensitive polymer alkaline phosphate prodrug micelles were designed and synthesized. The preparation of Copolymer Poly containing adamantane via RAFT polymerization (MPC-co-Ada), and Adamantane and adriamycin modified beta cyclodextrin confirmed by 2DNOESYNMR (DOX-hyd-CD) complex interaction between subject and object. The in vitro drug release results showed that the micelles prepared before Quick release can be realized by adriamycin in acidic conditions of hydrazone bond fracture simulation under the lysosomal / endosomal cells. Experimental results confirm the effective cell prodrug micelles endocytosis and inhibit the proliferation of HepG2 cells. Thanks to the "plug and play" object type for.3. of tumor therapy the prodrug micelles is also expected to further realize multiple functions of the preparation of phosphorylcholine based copolymers containing benzaldehyde groups further RAFT polymerization, and the hydrazone bond grafted onto the AIE molecule four phenyl ethylene (TPE) copolymer. The PMPC-hyd-TPE can self assemble to form with phosphorylcholine as shell with TPE micelles nuclear. TPE hydrophobic aggregation nucleation, but also can not only stimulate the AIE effect of doxorubicin loaded by hydrophobic interaction. The in vitro drug release experiments showed that in the acidic environment simulation cell, hydrazone The bond cleavage of TPE micelle swelling and rapid release of the adriamycin. The cell experiment showed that, the polymer micelles have AIE good imaging effect and can effectively inhibit the proliferation of HepG2 cells. At the same time, DLS results showed that phosphate containing FBS in micellar Bile Alkaline DMEM culture medium can maintain long-term stability, and plasma protein BSA and FBG were co incubated with 24h, the content of micelle adsorption protein remained at a low level. Further animal experiments of phosphocholine micelles can transfer.4. to adriamycin target tumor tissue to enrichment and to further explore the feasibility through simple polymerization step and a single near infrared dye load choline phosphate preparation treatment the integration of the homopolymer micelles. By alkyl chain length growth of phosphorylcholine monomer MPC obtained two amphiphilic monomer MDPC by one-step polymerization of RAFT can get two amphiphilic homopolymer PMDPC. the two amphiphilic homopolymer PMDPC can also self assemble to form stable micelle, and the effective load of near infrared dye IR-780. The homopolymer micelle load, light stability of near infrared dye IR-780 was significantly enhanced, and have high photothermal conversion effect, can effectively kill BxPC-3 pancreatic cancer cells. Animal experiment further, IR-780 loaded the phosphorylcholine homopolymer of micelles could effectively accumulate in the tumor region, with near infrared imaging of high resolution and long-term.

【學位授予單位】：浙江大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：R73-3;O631.3

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