奈米幾丁聚醣藥物載體之製備及應用研究=Preparation and application of Nano Chitosan drug carriers
曾詣文、
E-mail: [email protected]
摘 要
本研究目的在製備出奈米級的幾丁聚醣藥物載體，首先將市售幾丁聚醣溶於鹽酸中，並調整成適當之酸鹼值，乾燥後便得 可溶於水的幾丁聚醣鹽酸鹽。再將幾丁聚醣鹽酸鹽混和不同濃度之架橋劑-綠梔子素與消炎止痛藥-待克菲那配製成溶液，
進行噴霧乾燥得到奈米幾丁聚醣/待克菲那藥物載體，並對產物作粒徑及物性分析、細胞毒性、穿透實驗及藥物釋放之評 估。 由場發射電子顯微鏡觀察，各樣本平均粒徑約在300 nm。利用富立葉紅外線光譜儀偵測發現，樣本中含有藥物時，
在1456 cm-1處出現特徵吸收，且當架橋劑濃度增加時，在1540 cm-1之胺基吸收峰減小，顯示幾丁聚醣鹽酸鹽顆粒中有藥 物待克菲那的存在，與架橋劑有架橋反應生成。將藥物載體與腸道上皮細胞（Caco-2）進行細胞毒性及穿透評估，發現藥 物載體在濃度 100 ug/mL以下對Caco-2 細胞無明顯毒性。而由TEER實驗結果發現，樣本D有較佳的穿透能力達50 ％，若 樣本保存不善造成吸濕，顆粒變大則無法順利通過細胞間隙，細胞經過trypan blue染色後發現，樣本並未對細胞造成傷害
。由藥物釋放研究發現，在前1小時小於50 ％附著於表面的待克菲那會先被釋放出來，且隨時間增長藥物濃度也逐漸上升
，因此確認幾丁聚醣藥物載體具有緩釋藥物之能力。
關鍵詞 : 幾丁聚醣 ; 藥物載體 ; 噴霧乾燥 ; 藥物釋放
目錄
封面內頁 簽名頁 授權書... iii 中文摘要... iv 英文摘要... v 誌 謝... vi 目錄... vii 圖目錄... x 表目
錄... xii 1. 研究目的... 1 2. 文獻回顧... 2 2.1 幾丁質與幾丁聚醣簡 介... 2 2.2 幾丁聚醣鹽類... 2 2.3 架橋劑綠梔子素（genipin）簡介... 4 2.4 待克菲那
（diclofenac sodium）簡介... 7 2.5 幾丁聚醣於藥物控制釋放... 9 2.5.1 幾丁聚醣藥物載體的特性... 11 2.6 奈米化藥物載體... 13 2.6.1 噴霧乾燥法... 14 2.7 口服藥物傳輸模式... 16 2.7.1 Caco-2單層膜細胞... 16 2.7.2 幾丁聚醣物質於腸道穿透之研究... 17 2.8 細胞毒性評
估... 20 2.8.1 細胞毒性試驗方法... 20 2.9 藥物釋放評估... 22 2.9.1 分光光度計偵 測法... 22 2.9.2 HPLC定量法... 22 3. 材料與方法... 23 3.1 實驗架
構... 23 3.2 實驗材料... 24 3.2.1 藥品... 24 3.2.2 耗 材... 25 3.2.3 儀器設備... 26 3.2.4 細胞株... 27 3.3 研究方
法... 28 3.3.1 幾丁聚醣鹽酸鹽製備... 28 3.4 幾丁聚醣鹽酸鹽藥物載載體製備... 28 3.4.1 噴霧乾燥法製備奈米藥物載體... 30 3.5 產物分析... 30 3.5.1 場發射電子顯微鏡（FESEM
）... 30 3.5.2 富立葉紅外線光譜儀試驗（FTIR）... 31 3.6 細胞毒性之研究... 31 3.6.1 細胞復 甦... 31 3.6.2 繼代培養... 32 3.6.3 MTT毒性試驗... 34 3.7 細胞穿透之研 究... 34 3.7.1 TEER試驗... 35 3.7.2 Trypan blue細胞存活率試驗... 35 3.8 藥物釋放試 驗... 37 3.8.1 標準品配製... 37 3.8.2 藥物釋放分析... 37 4. 結果與討
論... 38 4.1 幾丁聚醣鹽酸鹽及載體之製備... 38 4.2 場發射電子顯微鏡觀察... 39 4.3 富 立葉紅外線光譜儀試驗（FTIR）... 52 4.4 細胞試驗... 57 4.4.1 Caco-2細胞培養觀察... 57 4.4.2 細胞毒性試驗（MTT分析）... 58 4.4.3 Tranepithelial electrical resistance （TEER）分析... 59 4.4.4 Trypan blue存活率試驗... 61 4.5 藥物釋放試驗... 68 5. 結論... 74 參考文 獻... 76
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