雙磷酸鹽類之骨質疏鬆藥物於新型可注射式溫感性載體之藥物釋放研究

Study on Bisphosphonates Drug for Osteoporosis in Novel Injectable Thermosensitive Carrier of Drug Delivery

周明潔(Ming-Chieh Chou)

指導教授 ： 葉瑞銘

中原大學/理學院/奈米科技碩士學位學程/碩士(2013年)

https://doi.org/10.6840/cycu201300873

摘要

中文摘要
本研究欲製得新型可注射式溫感性藥物緩釋載體，首先利用開環聚合方式合成具有溫度敏感性之高分子共聚合物(mPEG-PLGA)， 接著以化學共沉澱法合成氫氧基磷灰石，再利用漩渦混合器將兩者混摻，製成可注射性之溫感型複合水膠載體。然後使用發散型方式連續合成樹枝狀高分子藥物載台，並將 PAMAM G2 外圍改質為羧酸 (-COOH)，之後修飾上親水性鏈段 mPEG 合成 mPEG-PAMAM G2 ，以降低材料對細胞之毒性，並利用親疏水特性形成組裝體，包埋改質為Alendronate-N.Q.S之福善美(Alendronate)藥物，作為新型長效藥物載體，應用於骨質疏鬆症狀。
鑑定部分，水膠材料利用核磁共振頻譜儀 (NMR) ，傅利葉轉換紅外線光譜儀 (FTIR) 及凝膠滲透分析儀 (GPC) 等儀器對有機高分子 mPEG-PLGA 材料進行鑑定。無機氫氧基磷灰石的鑑定則利用 X-ray 繞射儀 (XRD)及傅利葉轉換紅外線光譜儀 (FTIR) ，判斷其結構正確性， X-RD實驗結果顯示在 2θ 角為 25.8° 處出現的波峰是氫氧基磷灰石主要的特徵峰，而其鈣/磷比亦與天然骨頭之成分相近。樹枝狀高分子材料部份，則利用核磁共振頻譜儀 (NMR)，傅利葉轉換紅外線光譜儀 (FTIR) 及四極柱分離飛行時間質譜 (TOF- MASS) 等儀器對樹枝狀高分子材料進行鑑定。

特性分析方面，分別對 mPEG-PLGA 、 mPEG-PLGA / HA 及 {mPEG-PLGA / HA}/ mPEG-PAMAM 之材料在溫度敏感性及體外降解行為研究探討，則藉由流變儀及體外降解實驗量測。體外毒性測試部分，以小鼠胚胎添加{mPEG-PLGA/ [mPEG-PAMAM-AL]}材料，24小時後用TUNEL 檢測法做測試，確定持續保有降低材料毒性之特性。最後，體外載藥量及藥物釋放量觀察，利用UV-vis 測量，改質接上發光基團 N.Q.S 後的 Alandronate-N.Q.S， 包覆於材料中之吸收值，代入檢量線算出包藥率、載藥量及各時間點的藥物釋放量，成功證實材料中的藥物得到緩釋效果。故由實驗結果得到以下結論，mPEG-PLGA/[mPEG-PAMAM G2-AL-N.Q.S]可製得緩釋注射式溫感性水膠，達到原位注射及藥物緩釋效果。

關鍵字

水膠 ； 聚醯胺樹枝狀高分子 ； 生物降解性.可注射性 ； 溫度敏感性 ； 毒性測試 ； 福善美 ； 藥物釋放

並列摘要

Abstract
In this research, we used vortex mixer to blend thermosensitive polymer (mPEG-PLGA) which is synthesized by ring-opening polymerization and hydroxyapatite by chemical co-precipitation to get complex carrier of thermosensitive injectable hydrogel. Then, we used dendrimer drug carrier, which is synthesized by stretch method continuous synthesis, to change the peripheral of PAMAM G2 to carboxylic acid (-COOH). For reducing the toxicity from materials to cells, we modified it with hydrophilic chain (mPEG) to get mPEG-PAMAM G2. After that, we used the characteristics of hydrophilic and hydrophobic for embedding revised Fosamax (Alendronate), to produce the carrier of the new long-acting drug, which can be applied in
The section of identification, we used nuclear magnetic resonance spectroscopy (NMR), Fourier transform infrared spectroscopy (FTIR), gel permeation chromatography (GPC) and other instruments to identify the organic polymer mPEG-PLGA. we used X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR) to identify and determine the correctness of its structure to the inorganic hydroxyapatite.In X-RD results are displayed at the 2θ angle of 25.8 ° occurring hydroxyapatite peak is the main peak , and its calcium / phosphorus ratio of composition are also similar to natural bone.Otherwise, we used nuclear magnetic resonance spectroscopy (NMR), Fourier transform infrared spectroscopy (FTIR), time-of-flight mass spectrometry (TOF-MASS) and other equipments for identification of dendrimer materials .
Characteristic analysis, rheometer and in vitro degradation measurements were used to detect the sensitivity to temperature and in vitro degradation behaviors of mPEG-PLGA, mPEG-PLGA / HA and mPEG-PLGA / mPEG-PAMAM respectively.
The last step, we examined the in vitro toxicity by injecting {mPEG-PLGA / [mPEG-PAMAM-AL]} into the embryo of the mouse and then left to stand for 24 hours before being tested by TUNEL detection. Finally, TUNEL detection confirmed that it’s successful to reduce the toxicity. The drug loading and the drug release were observed by UV-vis which can measure the absorption of the revised Alandronate with luminescent functional groups. Then, we integrated absorption values into calibration curve, to calculate the rate of drug wrapping, drug loading and drug release at various times. The conclusion shows that the long-acting thermosensitive injectable hydrogel, which was produced from mPEG-PLGA / [mPEG-PAMAM-AL-NQS], can reach the pharmaceutical sustained-release effect.

並列關鍵字

injectable ； biodegradability ； polyamide dendrimers ； Toxicity testing ； thermosensitive ； Fosamax ； hydrogel ； drug delivery

參考文獻

粒做為骨填補材料對骨修復之能力，國立陽明大學/醫學工程研究所/碩士

[1]K. Geier, (2001).Osteoporosis in men. Orthopaedic Nursing,20(6),49-56

[4] 蔡育霖，PLGA/Phospholipid 微小球對包覆之蛋白質控制釋放之影響，中原

[8] Tissue Engineering, August 2001, pages 441-455

[10] 施威任，奈米級氫氧基磷灰石之合成及燒結，國立成功大學/材料科學及