臺灣博碩士論文加值系統

此論文針對一系列新穎雙親性嵌段共聚高分子--- poly(ethylene glycol)-b-[polystyrene-co-poly(2-(1,2,3,4,5-pentaphenyl-1H-silol-1-yloxy)ethyl methacrylate)] [PEG-b-(PS-co-PAIE), P3]與poly(ethylene glycol)-b -[poly(2-nitrobenzyl methacrylate)-co-poly(2-(1,2,3,4,5-pentaphenyl-1H-silol-1 -yloxy)ethyl methacrylate)] [PEG-b-(PNBMA-co-PAIE), P4] 所自組裝形成的高分子微胞之奈米結構和光物理性質進行研究。我們以PEG做為親水鏈段，將一種具有Aggregation-Induced Emission (AIE) 性質的螢光側鏈基團引進高分子的疏水鏈段。在高分子側鏈的AIE基團可以克服傳統螢光染料在高分子聚集形成微胞時螢光被淬滅的問題。因此，具AIE特性之高分子所自組裝形成之微胞可作為螢光探針，用於追蹤微胞的位置，顯示其在生物標記具應用潛力。此外，P4具光裂解單體2- nitrobenzyl methacrylate (NBMA)，利用NBMA照光會裂解並從疏水性變為親水性的特性可將其應用於藥物控制釋放。
AIE基團在微胞核心的螢光強度高於溶解於有機溶劑中的螢光強度。證明微胞核心的狹小空間可以造成AIE基團的聚集，使AIE基團產生聚集誘發螢光。其中P3微胞的螢光比P4微胞的螢光強，因為NBMA的硝基會淬滅AIE螢光。藉由DLS測出P3與P4系列之高分子微胞粒徑約33.0 ~ 43.9 nm。另外，利用AIE特性可算出這兩系列高分子的臨界微胞濃度(Critical Micelle Concentration, CMC)約4.34×10-6至4.19×10-7 M。
P3與P4系列之高分子微胞進一步被用來包覆藥物 Doxorubicin (Dox)。由於AIE基團的螢光放射波長與Dox的吸收光譜重疊，當兩者之間距離小於10 nm時會產生Föster Resonance Energy Transfer (FRET)。此現象可確認Dox是否成功進入高分子微胞的核心。
經由在37℃ PBS水溶液中的藥物釋放實驗發現照光可促進P4/Dox微胞的Dox釋放。而in vitro實驗的部份則是以HT-29細胞株為對象，初步證實P3高分子微胞的生物相容性良好。未來期望能將P3與P4兩系列之高分子微胞都應用至in vitro的實驗，測試其在細胞內的生物標記與藥物釋放作用情形。

The nanostructures and photophysical properties of the fluorescent polymeric micelles self-assembled from a series of new amphiphilic block copolymers, poly(ethylene glycol)-b-[polystyrene-co-poly(2-(1,2,3,4,5-pentaphenyl-1H-silol-1 -yloxy)ethyl methacrylate)] [PEG-b-(PS-co-PAIE), P3] and poly(ethylene glycol)-b -[poly(2-nitrobenzyl methacrylate)-co-poly(2-(1,2,3,4,5-pentaphenyl-1H-silol-1 -yloxy)ethyl methacrylate)] [PEG-b-(PNBMA-co-PAIE), P4], were investigated in this work. We chose PEG as the hydrophilic block, and a fluorescent pendent group with the special characteristic of aggregation-induced emission (AIE) was introduced in the hydrophobic block of the copolymers. This AIE fluorescent moiety could overcome the problem of aggregation-induced quenching for most conventional dyes when encapsulated in the core of polymeric micelles. The chemical attachment of AIE moiety in the side chain of copolymers enables the emission of AIE moieties as the fluorescent probe to trace the location of polymeric micelles, suggesting its potential application in the bioimaging. In addition, the photocleavable monomers (2-nitrobenzyl methacrylate, NBMA) of P4 would show the transition from hydrophobic to hydrophilic after irradiated by UV light which could be used in the controlled drug release.
The photophysical properties of these fluorescent polymeric micelles were studied by the absorption and photoluminescence spectra. The emission from the AIE moieties in the core of micelles was observed with higher fluorescent intensity than the AIE moieties dissolved in the organic solvent. The nano confinement of AIE moieties in the core of micelles provides suitable environment for aggregation-induced emission. Unfortunately, the fluorescence of P4 was quenched by the nitro groups on NBMA, so the fluorescence of P4 was weaker than that of P3. The sizes of P3 and P4 series polymeric micelles were from 33.08 to 43.86 nm. The CMCs could be calculated about 4.34×10-6 to 4.19×10-7 M using the AIE effect.
The P3 and P4 series polymer were utilized as nanocarriers for encapsulation of cancer drug, doxorubicin (Dox). Due to the spectral overlap between the emission of AIE moieties and the absorption of Dox, the Föster Resonance Energy Transfer (FRET) from AIE moieties to Dox encapsulated in the micelles indicates the successful encapsulation of Dox in the core of polymeric micelles. According to the release profile of P4/Dox micelles in 37℃ PBS solution, the UV-irradiation could accelerate the release rate of Dox released from P4 micelles. The cytotoxicity test revealed the high biocompatibility of P3 micelles. In the future, we expect both P3 and P4 series polymeric micelles can be applied to in vitro experiments and investigate their application in bioimaging as well as intracellular drug release.

摘要 I
Abstract III
誌謝 V
目錄 VI
流程目錄 (LIST OF SCHEMES) IX
表目錄 (LIST OF TABLES) X
圖目錄 (LIST OF FIGURES) XI

第一章、緒論 1
1.1 研究背景與文獻回顧 1
1.1.1雙親性嵌段共聚高分子 1
1.1.2功能性高分子材料 3
1.1.2.1 螢光高分子 3
1.1.2.2 光敏性高分子 (photo-responsive polymers) 4
1.1.2.3 其他功能性高分子 6
1.1.3 螢光材料 6
1.1.3.1螢光 6
1.1.3.2 螢光材料 9
1.1.3.3 Aggregation-induced Emission (AIE) 螢光材料 12
1.1.3.4 Föster Resonance Energy Transfer（FRET） 15
1.1.4藥物傳輸與癌症治療 16
1.1.4.1 Enhanced Permeation and Retention (EPR) 效應 17
1.1.4.2 藥物載體 18
1.1.4.3 光動力治療 19
1.2 研究動機與目的 20

第二章、實驗 21
2.1 實驗藥品 21
2.2 實驗方法 23
2.2.1 單體合成 23
2.2.1.1 AIE單體 2-(1,2,3,4,5-pentaphenyl-1H-silol-1-yloxy)ethyl methacrylate (PPS-HEMA) 23
2.2.1.2光裂解單體2-nitrobenzyl methacrylate (NBMA) 24
2.2.2 高分子聚合 24
2.2.2.1 [α-(2-bromo-2-methylpropoyloxy)]-poly(ethylene glycol) (PEG5000 macroinitiator) 24
2.2.2.2 poly(ethylene glycol)-b-polystyrene (PEG-b-PS) 25
2.2.2.3 poly(ethylene glycol)-b-poly(2-nitrobenzyl methacrylate) (PEG-b-PNBMA) 26
2.2.2.4 poly(ethylene glycol)-b-[polystyrene-co-poly(2-(1,2,3,4,5-pentaphenyl-1H-silol-1-yloxy)ethyl methacrylate)] (PEG-b-[PS-co-P(PPS-HEMA)]) 27
2.2.2.5 poly(ethylene glycol)-b-[poly(2-nitrobenzyl methacrylate)-co-poly(2-(1,2,3,4,5-pentaphenyl-1H-silol-1-yloxy)ethyl methacrylate)] (PEG-b-[PNBMA-co-P(PPS-HEMA)]) 28
2.2.3 微胞製備與CMC測量 29
2.2.4 光裂解測試 29
2.2.5 藥物包覆 30
2.2.6 藥物釋放 31
2.3 儀器鑑定 31
2.3.1 Gel permeation chromatography (GPC) 31
2.3.2 Nuclear Magnetic Resonance (NMR) 32
2.3.3 Dynamic Light Scattering (DLS) 32
2.3.4 Transmission Electron Microscopy (TEM) 33
2.3.5 Ultraviolet-visible spectra (UV-vis. spectra) 33
2.3.6 Fluorescence spectra and Quantum Yield (Q.Y.) 34

第三章、結果與討論 35
3.1 聚合和鑑定 35
3.2 奈米結構 43
3.3 光物理性質 44
3.4 光裂解測試 50
3.5 藥物包覆 54
3.5.1 測試P3系列高分子包覆藥物之能力與特性 54
3.5.2 測試P4系列高分子包覆藥物之能力與特性 56
3.6 藥物釋放 59
3.7 細胞毒性 62

第四章、結論與未來工作 63

第五章、參考文獻 65

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