改質後幾丁聚醣/硫酸化軟骨素聚電解質錯合物作為蛋白質藥物載體之研究

Modified Chitosan and Chondroitin Sulfate Complexes for Protein Drug Delivery

王柏勝(Bo-Sheng Wang)

高雄醫學大學/生命科學院/醫藥暨應用化學研究所/碩士(2005年)

摘要

近年來利用聚電解質錯合物來作為藥物載體或是控制釋放系統已經被廣泛的研究。本研究利用化學改質製備幾丁聚醣（Chitosan, Chi）溶於醋酸水溶液中，形成完全水溶性後再與硫酸化軟骨素（Chondroitin sulfate, ChS）形成聚電解質錯合物，作為蛋白質藥物包覆載體，並觀察其釋放行為。我們將分子量330,000 g/mole的Chi利用NaBO3水解濃度及反應時間得到分子量90,000與40,000 g/mole的幾丁聚醣，並以EX-810（Ethylene glycol diglycidyl ether）作為間連劑 (spacer) 接枝不同莫耳數含量之（Polyethyleneimine, PEI）簡稱為PEI-g-Chi。由1H-NMR來分析改質後幾丁聚醣之去乙醯化程度D.D.（Degree of deacteylation）值和接枝率。PEI-g-Chi與硫酸化軟骨素形成錯合物，殘留之-NH2基團的量由Ninhydrin assay來檢測。在磷酸鹽緩衝溶液（PBS buffer, pH=7.4）中， Chi錯合物對BSA的釋放量偏低，可能因為錯合物對BSA的作用力很強；另外，PEI-g-Chi的錯合物釋放BSA的量更加的少，顯示出改質後的幾丁聚醣形成之錯合物提高了對BSA的結合力。

關鍵字

去乙醯化程度 ； 寧海分析

並列摘要

Polyelectrolyte complexes have been widely used for drug-controlled release in recent years. The polyelectrolyte complexes between Chitosan (Chi) and Chondroitin Sulfate (ChS) were used as a protein drug carrier and their sustain-released behavior depending on the molecular weight of Chitosan was studied. The Mv=330,000 g/mole of parent chitosan was cut into Mv=90,000g/mole and Mv=40,000g/mole with NaBO3 by controlling reacting time. Furthermore, the various molar ratios of PEI (polyethyleneimine) were grafted with chitosan using EX-810 (Ethylene glycol diglycidyl ether) as a spacer as well as a conjugation reagent. The DD (Degree of deacetylation) and DS（Degree of substitution）of modified chitosan were checked by 1H-NMR. The free amine groups of Chitosan and PEI-g-Chi were quantitatively determined by ninhydrin assay. The sustained release of BSA from the complexes was tested in PBS buffer at pH 7.4. The results show that the BSA released form complexes are not conspicuous, indicating that the interactions between BSA with complexes are strong. In addition, the BSA released from the complexes prepared from PEI-g-Chi is less than its respective parent chitosan, suggesting that the modified Chi indeed enhance the binding ability to BSA.

並列關鍵字

drgree of deacetylation ； ninhydrin assay

參考文獻

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