臺灣博碩士論文加值系統

己醛醣酸鹽水解酵素為一種醣水解酵素，其主要的功能為水解葡萄糖胺聚醣中硫酸乙醯肝素或皮膚素硫酸鹽末端的艾杜醣醛酸。此酵素的缺陷將造成第一型黏多醣症。但對研究己醛醣酸鹽水解酵素之文獻仍不完備。如利用化學合成方式取得己醛醣酸鹽水解酵素螢光受質，步驟繁瑣與產率不佳。以及在酵素抑制劑設計上的不明確。本論文的目的為改善現有人工合成、合成方法之不足處與探討酵素抑制劑的設計。
在己醛醣酸鹽水解酵素受質的製備，我們參考現有文獻，以3,6-丁內酯-D-葡萄喃醣為起始物。經由甲氧基開環與乙醯基化，再經由溴化反應以自由基反應將五號位置的立體組態翻轉，取得上保護的L-艾杜醣關鍵中間體。透過一號位置選擇性水解與醣基化反應，來取得己醛醣酸鹽水解酵素受質。
另一方面在己醛醣酸鹽水解酵素抑制劑的設計，由酵素受質與抑制劑的結構差異，讓我們在抑制劑的設計上，由抑制劑的構型上的差異、一號位置的位向以及一號位置的衍生化設計，來比較其分子結構與活性的關係。
己醛醣酸鹽水解酵素受質的合成目前可合成百毫克的受質，且實際應實際應用於本文中之生物活性測試上。在抑制劑的探討，以1C4的構型的艾杜醣酸也可當作抑制劑設計的骨架。藉由[3 + 2] cycloaddition來將此骨架在一號位置衍生化。所得到最具抑制效果化合物在100µM下具50% 抑制活性。此初步成果可提供未來化學分子設計之基礎。

Currently, the fluorescent substrate and inhibitors study of α-L-iduronidase were rarely mentioned in literatures. The purpose of this work is to improve the methods of synthesis of α-L-iduronidase fluorescent substrate and to design α-L-iduronidase inhibitors. In this work, inspired by known inhibitors and α-L-iduronidase substrates, we also synthesized new inhibitors and elucidated the conformation, configuration and C1 diversity of inhibitors to investigate the structure and activity relationship.

We have successfully and efficiently developed a convenient method to prepare about α-L-iduronidase fluorescent substrate in hundred milligram scalp, with which to study the enzyme and cell based inhibition assay. In terms of inhibitors, the L-iduronic acid with 1C4 conformation can be a potential scaffolds. By employing [3+2] cycoaddition to modify C-1 substituent to give inhibitors which showed 50% enzyme inhibition at 100 µM. This preliminary study provides the basis on designing α-L-iduronidase inhibitors in the future.

摘要 I
Extended Abstract II
誌謝 VI
目錄 VII
表目錄 X
圖目錄 X
流程目錄 XII
中英文對照表 XIII
簡稱語對照表 XIV
第一章 緒論 1
1.1溶小體儲積症 (Lysosomal storage disease): 1
1.2黏多醣貯積症介紹 (Mucopolysaccharidosis): 2
1.3 第一型黏多醣儲積症治療方式: 4
1.4葡萄糖胺聚醣 (Glycosaminoglycan) 5
1.5己醛醣酸鹽水解酵素α-L-Iduronidase (IDUA): 6
1.6己醛醣酸鹽水解酵素(α-L-Iduronidase)受質 7
1.6.1 Baggett的合成: 9
1.6.2 Hung的合成: 10
1.6.3實驗室先前發展之合成: 11
1.7己醛醣酸鹽水解酵素 (α-L-iduronidase) 抑制劑 12
1.7.1抑制劑(5F-IdoA)與酵素的共結晶結構關係 13
1.7.2五元環亞胺醣抑制劑 14
1.8 研究動機: 15
1.8.1合成己醛醣酸鹽水解酵素受質 15
1.8.2合成己醛醣酸鹽水解酵素抑制劑 15
第二章 結果與討論 16
2.1己醛醣酸鹽水解酵素受質逆合成分析 16
2.1.1溴化物4的製備 17
2.1.2化合物3的製備 18
2.1.3化合物2的製備 21
2.1.4化合物1的製備 22
2.1.5 己醛醣酸鹽水解酵素受質的定量與生物測試 23
2.2己醛醣酸鹽水解酵素抑制劑目標產物設計概念 25
2.2.1己醛醣酸鹽水解酵素抑制劑之逆合成分析 26
2.3 己醛醣酸鹽水解酵素抑制劑合成討論 28
2.3.1 Azide 8的合成 28
2.3.2 路徑一化合物7的合成 29
2.3.3 路徑一化合物5F- IdoA-R的合成 31
2.3.4 路徑二化合物 5F- IdoA-R的合成 31
2.3.5 路徑四化合物 5F-IdoA-R的合成 33
2.3.6 1C4構型艾杜醣酸衍生物化合物3α製備 34
2.3.7 化合物9α製備 34
2.3.8 化合物IdoA-αR的製備 36
2.3.9 化合物IdoA-βR的製備 37
2.4 艾杜糖酸衍生物生物活性測試 38
2.4.1 化合物IdoA-αR的製備 40
2.4.2 不同取代基性質的苯環對抑制效果的影響 43
2.5 結論 44
第三章 實驗部份 45
3.1 實驗藥品及溶劑： 45
3.2 實驗儀器： 46
3.3 實驗步驟與光譜數據 46
第四章 參考文獻 71
附錄 75

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