臺灣博碩士論文加值系統

本研究係針對三組具特殊功能之雙親性嵌段共聚高分子Poly(ɛ-caprolactone)-b-poly[triethylene glycol methacrylate-co- 6-(methacrylamido)hexanoic acid] [PCL-b-P(TEGMA-co-AHA)]、Poly(ɛ-caprolactone)-b-poly[triethylene glycol methacrylate-co- N-(2-(methacrylamido)ethyl) folatic amide] [PCL-b-P(TEGMA-co-FA)]與Poly(ɛ-caprolactone)-b-poly[triethylene glycol methacrylate-co- (2-(1,2,3,4,5-pentaphenyl-1H-silol-1-yloxy)ethyl methacrylate)] [PCL-b-P(TEGMA-co-(PPS-HEMA))] 一起於水溶液中自組裝形成之高分子混合微胞進行奈米結構、環境應答、螢光性質、標靶功能及此藥物載體系統對細胞毒性之探討與檢測。
PCL作為本系統中疏水鏈段構成微胞內核，用以包覆疏水性藥物(Doxorubicin, DOX)，PTEGMA作為親水鏈段微胞外殼，用以增加微胞於水溶液中的穩定性並使高分子具有溫度響應性質。我們分別在三種組成混合微胞的高分子親水鏈段修飾上酸鹼敏感性單體AHA、具主動標靶功能的FA以及具有螢光特性單體PPS-HEMA，使各組雙親性嵌段共聚高分子具有特定的功能。
透過調整具酸鹼值響應單體AHA於親水鏈段中的比例及各組高分子於混合微胞中組成的比例可以調控高分子及混合微胞的最低臨界溶解溫度(lower critical solution temperature, LCST)，使其達到於中性環境時LCST高於人體體溫37℃，於酸性環境時LCST低於37℃，因此在中性環境的血液循環中乘載藥物的混合微胞可以穩定存在，而當微胞進入酸性環境之癌細胞溶酶體時，微胞結構會因親疏水性的改變而被破壞使所乘載的藥物釋放。高分子上所修飾上的葉酸(FA)可與癌細胞表面的葉酸受體結合，使藥物能更有效地在目標細胞累積，我們修飾上不同比例的FA來探討FA對癌細胞標靶傳遞及細胞毒性的影響；而PPS-HEMA是具AIE性質的螢光物質，其放射波長與DOX的吸收波長重疊，因此當兩者距離夠近時會產生Föster Resonance Energy Transfer (FRET)效應，我們利用FRET作為判斷藥物是否成功包覆於微胞中及包覆量多寡的依據。

In this study, we prepare a mixed micelle system as drug carriers. The mixed micelles were co-assembled of three amphiphilic copolymers, [PCL-b-P(TEGMA-co-FA)], [PCL-b-P(TEGMA-co-AHA)] and [PCL-b-P(TEGMA-co-(PPS-HEMA))]. PCL block is hydrophobic and used as the core of micelle to encapsulate hydrophobic drug, DOX. PTEGMA block is thermo-sensitive and acts as the hydrophilic shell of micelle. In addition, we introduced the pH-sensitive, active targeting and fluorescent monomer (AHA, FA and PPS-HEMA) to the hydrophilic block by copolymerization the corresponding comonomers with TEGMA, respectively. The nanostructure, stimuli-responsive properties, active targeting properties, florescence behaviors and cytotoxicity for tumor cells were investigated. With delicate control of the composition of AHA in the hydrophilic block, the LCST of mixed micelles exhibited pH-dependent thermal transition behaviors suitable for drug delivery systems that LCST is higher than body temperature (37℃) at neutral environment but lower than body temperature at acidic environment. Therefore, the drug can be encapsulated in the core of micelle stably in blood circulation and released in tumor which is acidic because the micellar structure is destroyed. FA can conjugate with the targeted tumor with FA receptors overexpressed on the surface to make drug accumulate more efficiently. PPS-HEMA is a florescence monomer with AIE effect, and FRET will occur when the spectral overlap and close proximity between PPS-HEMA and DOX. Based on FRET effect, we are able to analyze if DOX was successfully encapsulated in the mixed micelle.

摘要 I
目錄 XIII
流程圖目錄 XV
表目錄 XVI
圖目錄 XVII
第1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(Amphiphilic block copolymers,ABCs) 1
1.1.2雙親性嵌段共聚高分子於藥物傳遞之應用 5
1.1.3 微胞製備 14
1.1.4功能性高分子 15
1.1.5 混合微胞(Mixed micelles) 30
1.2研究動機與目的 31
第2章、實驗 33
2.1實驗藥品 33
2.2實驗方法 36
2.2.1 單體合成 36
2.2.2 高分子聚合 39
2.2.3 微胞製備 46
2.2.4 Lower critical solution temperature (LCST)測試 46
2.2.5藥物包覆與釋放 46
2.3儀器鑑定 50
2.3.1 Gel permeation chromatography (GPC) 50
2.3.2 Nuclear Magnetic Resonance (NMR) 51
2.3.3 Dynamic Light Scattering (DLS) 51
2.3.4 Ultraviolet-Visilbe Spectroscopy (UV-vis.) 52
2.3.5 Photoluminescence Spectroscopy (PL) 52
第3章、結果與討論 53
3.1聚合與鑑定 53
3.1.1 Hydroxyethyl 2-bromoisobutyrate (HEBiB)合成 55
3.1.2 TEGMA單體合成 55
3.1.3 NSMA單體合成 56
3.1.4 PPS-HEMA單體合成 57
3.1.5 Poly(ɛ-Caprolactone)(PCL，P0)聚合 58
3.1.6雙親性嵌段共聚高分子poly(ɛ-caprolactone)-b-poly[triethylene glycol methacrylate-co- N-hydroxysuccinimide methacrylate] [PCL-b-P(TEGMA-co-NSMA)，P1]之合成 61
3.1.7雙親性嵌段共聚高分子 [PCL-b-P(TEGMA-co-AHA)](P1-AHA)、 [PCL-b-P(TEGMA-co-FA)] (P2-FA、P3-FA)之合成 64
3.1.8雙親性嵌段共聚高分子[PCL-b-P(TEGMA-co-(PPS-HEMA))] (P4-AIE)之合成 68
3.2混合微胞的製備與性質鑑定 71
3.2.1溫度敏感性質 72
3.4藥物包覆與釋放測試 84
3.4.1藥物包覆 84
3.4.2藥物釋放 88
3.5細胞毒性測試 91
第4章、結論與未來工作 95
第5章、文獻回顧 97

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