臺灣博碩士論文加值系統

近年來，奈米載體作為藥物傳遞系統已成為熱門的研究課題，而眾多的材料當中又以智慧型奈米載體最受矚目，當載體受到周圍環境(溫度，pH值，光或酵素)改變或作用時，會造成載體親疏水性改變或經化學催化反應來達到藥物釋放的效果。另一方面，光動力治療(photodynamic therapy， PDT) 是一種相當新穎有效且具選擇性的癌症治療方法，其主要是利用光動力治療所產生的單重態氧，造成腫瘤組織的凋亡。
本論文利用原子轉移自由基聚合與開環聚合法，成功製備出雙親性嵌段共聚高分子Poly(ε-Caprolactone)-b-poly[N-isopropylacrylamide- co-(N-methacryloyl-β-alanine)][PCL-b-P(NIPAAm-co-βA)]進行研究，探討自組裝形成高分子微胞之刺激應答性質、奈米結構及作為藥物載體之應用。我們選用具生物可分解性的PCL作為疏水鏈段，以此作為包覆疏水性抗癌藥物及光敏藥物之微胞內層，再配合具有溫度應答的PNIPAAm作為微胞外殼，及具官能化集團的N-hydroxysuccinimide methacrylate (NSMA)，經由化學修飾NSMA鏈段，形成具有酸鹼應答性質的β-alanine基團。藉由調整不同比例的NIPAAm以及β-alanine可有效控制其應答溫度及pH變化範圍。
PCL-b-P(NIPAAm-co-βA)系列高分子，在尚未修飾成酸鹼應答鏈段時，其LCST於中性環境中為28~29℃，酸性環境(pH 6.01)下則為26℃左右。經修飾後，在不同酸鹼環境下，由於β-alanine上羧酸去質子化程度不同，改變原本的親疏水平衡，在pH 6.01其LCST為25~32℃之間，在pH 7.05其LCST可高於37℃。在藥物釋放實驗中，PCL-b-P(NIPAAm-co-βA)在酸性環境下釋放速率較中性環境快，說明此載體具有雙重應答之特性。以子宮頸癌細胞(HeLa calls)作細胞毒性測試，其結果顯示此共聚高分子所形成之微胞具有良好的生物相容性。此外，本研究中製備的雙親性共聚高分子成功的作為藥物載體，除了包覆抗癌藥物Dox還結合了光動力治療。當包覆了光敏藥物之微胞，在特定光源之照射下，光敏藥物可有效產生具細胞毒性的單重態氧。將此微胞運用於藥物載體上，除了一般的抗癌藥物治療效果，還配合光動力治療，使其可達到雙重治療的效果。綜合此研究結果，以PCL-b-P(NIPAAm-co-βA)在做為控制藥物釋放之藥物載體上有良好之效果。

The utilization of drugs delivery system to treat tumors have been widely studided in recent years. Intelligent materials that respond to specific stimuli, such as temperature, pH, or enzymatic activity have been attracted great attention. In addition, the photodynamic therapy (PDT) is a novel and noninvasive technique for certain kinds of cancer tumor treatment, which utilized photosensitizer, oxygen and light together to generate singlet oxygen to induce apoptosis of tumor tissue.
Herein, we use atom transfer radical polymerization (ATRP) and ring opening polymerization (ROP) to synthesize a series of amphiphilic block copolymers, Poly(ε-Caprolactone)-b-poly[N-isopropylacrylamide- co-(N- metha-cryloyl-β-alanine)] [PCL-b-P(NIPAAm-co-βA)], and these copolymers can self-assemble to form micellar structures. We chose biodegradable polycaprolactone (PCL) as hydrophobic site, and thermo-sensitive polymer N-isopropylacrylamide (NIPAAm) and a reactive functional N-hydroxysuccinimide methacrylate (NSMA) as hydrophilic site. By post-funcionalization of β-alanine with NSMA moiety, and these copolymers PCL-b-P(NIPAAm-co-βA) will have pH-sensitive property. These dual stimuli-responsive copolymers have phase transition behavior in different pH aqueous solution, which can be manipulated by adjusting the mole ratio between NIPAAm and β-alanine.
In this work, PCL-b-P(NIPAAm-co-NSMA) phase transition behavior is independent with pH-responsive, and with introduction of β-alanine, PCL-b-P(NIPAAm-co-βA) have showed pH dependent lower critical solution temperature(LCST). Furthermore, we investigate release profile of the polymer micelles loaded with anti-cancer drug, doxorubicin (Dox) in both natural and acidic condition at 37℃. At natural condition, the micelles showed a slower release rate than that of acid, indicating that the micelles are more stable in natural condition, and at acidic condition the drug will release fast, so that they can be used in controlled drug release system. Beside, we also encapsulated the photosensitizer for PDT applications. Under light irradiation, photosensitizer transfers its energy to surrounding oxygen to generate cytotoxic singlet oxygen. This study indicates that our micelles can simutaneously incorporate two kinds of drug for chemotherapy and PDT and have the potential to applied as drug delivery system.

中文摘要 I
Abstract III
目錄 V
表目錄 VIII
圖目錄 X
第一章、 緒論 1
1-1研究背景與文獻回顧 1
1-1-1光動力治療之簡介 1
1-1-2光動力治療之機制 2
1-1-3光動力治療的三大要素 3
1-1-3.1光源 4
1-1-3.2 氧氣、光敏藥物 5
1-1-4光動力治療引發細胞死亡機制 11
1-2藥物傳輸系統 14
1-2-1 藥物載體(drug carriers) 15
1-2-2藥物輸送 28
1-2-2-1腫瘤組織與奈米藥物載體之關係 28
1-2-2-2奈米藥物載體在癌症治療之傳遞模式 31
1-2-3奈米藥物載體釋放 35
1-3環境應答型高分子 35
1-3-1溫度應答型高分子(thermosensitive polymer) 36
1-3-2酸鹼應答型高分子(pH-sensitive polymer) 41
1-4研究動機與目的 47
第二章、實驗藥品及方法 48
2-1實驗藥品 48
2-2實驗方法 50
2-2-1單體合成 50
2-2-1-1 N-hydroxysuccinimide methacrylate(NSMA) 50
2-2-1-2 Hydroxyethyl 2-bromoisobutyrate (HEBiB initiator) 50
2-2-2高分子合成 51
2-2-2-1 Poly(ε-Caprolactone)(PCL)聚合 51
2-2-2-2Poly(ε-Caprolactone)-b-poly[N-isopropylacrylamide-co- N-hydroxysuccinimide methacrylate][PCL-b-P(NIPAAm-co-NSMA)] 52
2-2-2-3Poly(ε-Caprolactone)-b-poly[N-isopropylacrylamide-co-(N-methacryloyl-β-alanine)][PCL-b-P(NIPAAm-co-βA)] 54
2-2-33 Britton-Robinson (B-R) 緩衝溶液 55
2-2-4高分子微胞製備 55
2-2-5 Critical micelle concentration (CMC)檢測 56
2-2-6 Lower critical solution temperature (LCST)測試 56
2-2-7藥物包覆 57
2-2-8 Chlorin產生單重態氧檢測 59
2-2-9藥物釋放 60
2-2-10細胞毒性 60
2-3 儀器鑑定 61
2-3-1 Nuclear magnetic resonance (NMR) 61
2-3-2 Gel permeation chromatography (GPC) 61
2-3-3 Dynamic light scattering (DLS) 62
2-3-5 Transmission electron microscopy (TEM) 62
2-3-6紫外-可見光吸收光譜(UV-vis.) 62
第三章、結果與討論 64
3-1聚合與鑑定 64
3-1-1NSMA單體合成 65
3-1-2 Hydroxyethyl 2-bromoisobutyrate(HEBiB initiator)起始劑合成 66
3-1-3 Poly(ε-Caprolactone)(PCL)聚合 67
3-1-4雙親性嵌段共聚高分子Poly(ε-Caprolactone)-b-poly[N-isopropylacrylamide-co-N-hydroxysuccinimide methacrylate][PCL-b-P(NIPAAm-co-NSMA)，P1~P3]之合成 69
3-1-5雙親性共聚高分子Poly(ε-Caprolactone)-b-poly[N-isopropylacrylamide-co-(N-methacryloyl-β-alanine)][PCL-b-P(NIPAAm-co-βA)]之合成 71
3-2微胞製備與性質鑑定 72
3-2-1 臨界微胞濃度(CMC)量測 73
3-2-2Lower critical solution temperature量測 75
3-2-3 DLS粒性分析 81
3-3藥物包覆與釋放測試 83
3-3-1藥物包覆 83
3-3-2光動力治療之單重態氧產生檢測 89
3-3-3藥物釋放 91
3-3-4細胞毒性 95
第四章、結論與未來工作 97
第五章、參考文獻 98

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