- 學位論文
葡聚醣接枝月桂酸高分子微胞搭載IKK複合體抑制劑應用於黑色素瘤細胞凋亡探討
The Study of Dextran graft Lauric Acid Polymeric Micelles Loaded IKK Complex Inhibitor Induce Cancer Cell apoptosis
摘要
摘要 文獻記載IKK複合體抑制劑(I kappa B kinase inhibitor，又名IMD-0354)具有治療黑色素瘤之潛力。本研究以天然葡聚醣與月桂酸完成之微胞可做為該藥物之載體。在方法上，是以酯化反應製備Dex-LA (Dextran-Lauric Acid)微胞載體，此微胞之化學結構與接枝率使用傅立葉轉換紅外線光譜儀和液態核磁共振儀鑑定，以動態雷射散射儀與穿透式電子顯微鏡測定微胞粒徑大小及形態，結果顯示微胞的形態為球形。搭載含有藥物IMD-0354之微胞載體IMD-Dex6-LA25、IMD-Dex6-LA50、IMD-Dex40-LA25和IMD-Dex40-LA50，其粒徑大小分別為63.94、90.55、103.36和67.83奈米，IMD-0354藥物包覆率分別為38.1、93.75、15.4和39.46 %，載藥率分別為1.9、4.67、0.76和1.96 %。IMD-Dex6-LA50微胞於磷酸鹽緩衝溶液(pH 7.4)中72小時之藥物釋放量為86.15 %。纖維母細胞L929進行試驗結果顯示Dex6-LA50微胞具有生物相容性，由共軛焦顯微鏡證實癌細胞B16F10胞飲螢光微胞FITC-Dex6-LA50。細胞存活率探討結果顯示癌細胞B16F10分別與藥物IMD-Dex6-LA50和IMD-0354共培養72小時的半致死劑量(即Lethal Dose, 50%)分別為0.43及0.57 μg/mL。由免疫螢光染色細胞核探討細胞凋亡過程細胞核轉變的實驗結果，顯示培養3天後之癌細胞B16F10顯著凋亡，證實IMD-0354能夠促使癌細胞B16F10走向凋亡。總此，本研究成功製備天然高分子微胞載體可用以搭載疏水性藥物，在未來有潛力應用於腫瘤之治療。
並列摘要
Abstract Some literatures reported that IKK inhibitor (I kappa B kinase inhibitor, known as IMD-0354) has potential to treat melanoma. This study synthesized Laurate-g-Dextran (Dex-LA) micelles to be used as a drug carrier for the IKK inhibitor. The synthesis is by esterifying the natural polymers Dextran and Lauric acid. By varying the weight of Dextran and Lauric acid, the micelles Dex6-LA25, Dex6-LA50, Dex40-LA25 and Dex40-LA50 were formed. The structure and the graft ratio of these Dex-LA micelles were characterized by Fourier transform infrared spectroscopy and 1H nuclear magnetic resonance, respectively. The particle size and the morphology of the micelles were confirmed by dynamic light scattering and transmission electron microscope (TEM), respectively. The TEM images showed that the micelles morphology is spherical. The particle size of the IMD-0354 encapsulated Dex-LA micelles (IMD-Dex-LA) is in the range of 63.94~103.36 nm. In these IMD-Dex-LA micelles, the encapsulation efficiency of IMD-0354 is from 15.4 to 93.75%, and the drug loading content is in the range between 0.76 and 4.76%. In addition, the cumulative drug release percentage of micelles is 86.15% in the phosphate buffer solution (pH 7.4) after 72 hours. The study of in vitro cytotoxicity showed that the Dex-LA micelles is biocompatible with the cell line of L929. Melanoma cells (B16F10) that endocytosed the FITC-labeling fluorescent micelles were confirmed by confocal laser microscopy. Moreover, the results of B16F10 cell viability study indicated that the LD50 (Lethal Dose, 50%) of IMD-0354 and IMD-Dex-LA is 0.43 and 0.57 μg/mL, respectively, for 72-hours co-culture of the B16F10 with the drugs. In the cellular apoptosis assay study, the results demonstrated that the death of the B16F10 is IMD-0354-induced apoptosis. In summary, this study successfully synthesized natural polymer-based micelles which can be used as a carrier for hydrophobic drug; and that it is believed the micelles has potential in the cancer treatment applications.