指導教授：羅禮強臺灣大學：化學研究所奚偉恩Hsi, WayneWayneHsi2014-11-252018-07-102014-11-252018-07-102013http://ntur.lib.ntu.edu.tw//handle/246246/261260己醛醣酸鹽水解酵素為內切型醣水解酵素，它的功能在於切除葡萄糖胺多醣體內L-艾杜醣,D-葡萄糖胺之間的1,4-醣苷鍵。已知文獻中鮮少提及己醛醣酸鹽水解酵素的抑制研究，因為L-艾杜醣，L-艾杜醣酸相關的酵素受質難以從天然物取得，需要人工合成，價格昂貴。因此有必要開發新方法取得己醛醣酸鹽水解酵素受質，以降低成本、自給自足。 己醛醣酸鹽水解酵素是罕見遺傳疾病「黏多醣儲積症第一型」的致病酵素。 我的合成路徑從1,2:5,6-雙異亞丙基-α-D-五環葡萄糖，經五號碳立體組態翻轉取得3-OBn-L-艾杜醣的關鍵中間體，透過三氯乙酰亞胺酯與4-甲基-7-羥基香豆素作醣化反應，TEMPO選擇性氧化一級醇，取得了己醛醣酸鹽水解酵素的螢光受質。 另一方面，為模仿此酵素進行醣水解反應時的過渡態，設計了兩個二號碳帶亞甲基胺的吡咯啶胺醣骨架，從五圓環硝中間體，經氮原子保護和TEMPO/NaOCl/NaClO2選擇性氧化，合成五圓環硝酸鹽。最後在其中一個吡咯啶胺醣骨架透過醯胺鍵接上一個酸，以用於後續酵素抑制性研究。α-L-iduronidase (IDUA), an endoglycosidase, cleaves the 1,4-glycosidic bond between L-iduronic acids (L-IdoA) and D-glucosamine (or D-galactosamine) in heparin and heparan sulfate. The inhibition study of IDUA is important but rarely mentioned in the literature because materials such as L-Idopyranose and L-IdoA are not commercially available. And substrates are expensive. Thus, an economical source of IDUA substrate is needed. Currently, IDUA has been found to involve with a genetically inherited lysosomal storage disease, mucopolysaccharidosis I (MPS1). A synthetic route started from 1,2:5,6-di-O-isopropylidene-α-D-glucofuranose, through C-5 inversion of configuration gave a key intermediate 3-OBn-L-idopyranose, followed by trichloroacetimidate-mediated glycosylation with 4-methylumbelliferone and selective primary alcohol oxidation by TEMPO catalyst to give the 4-methylumbelliferyl α-L-iduronide (4MU-α-IdoA) as a fluorogenic substrate for IDUA. To mimic the oxocarbenium ion generated during IDUA hydrolysis, 2-(aminomethyl) pyrrolidine iminosugar scaffolds were prepared using a five-membered chiral cyclic nitrone as a key intermediate, followed by N-protection and selective oxidation by TEMPO/NaOCl/ NaClO2 to give the aza-L-iduronic acid analogs. An acid was coupled with a scaffold to. diversify at primary amine as a model for further inhibition study.目 錄 口試委員會審定書………………………………………………………………………i 序言與致謝………………………………………………………………………………ii 中文摘要…………………………………………………………………………………iii Abstract………………………………………………………………………………iv Table of Contents…………………………………………………………………………v Index of Figures…………………………………………………………………………vii Index of Tables………………………………………………………………………viii Index of Schemes…………………………………………………………………………ix Abbreviations………………………………………………………………………………x Chapter 1. Introduction……………………………………………………………………1 1.1 Introduction of α-L-iduronidase (IDUA)…………………………………………1 1.2 Introduction of Mucopolysaccharidosis type I…………………………2 1.3 Natural substrate of IDUA…………………………………………………………5 1.4 Artificial substrates of IDUA…………………………………………………………6 1.5 Review of current IDUA inhibitors………………………………………………9 1.6 Motivations……………………………………………………………………10 Chapter 2. Results and discussion…………………………………………………12 2.1 Preparation of 4MU-α-IdoA by standard protection and deprotection route………12 2.1.1 Preparation of compound 8………………………………………………12 2.1.2 Preparation of compound 11………………………………………………14 2.1.3 Preparation of compound 1…………………………………………………15 2.2 Modified preparation route of 4MU-α-IdoA by late stage oxidation……………17 2.3 Azafuranose based transition state analogues……………………………………21 2.3.1 Preparation of azafuranose scaffold……………………………………21 2.3.2 Preparation of 2 and 3……………………………………………………24 2.3.3 1H NMR NOESY spectrums of 2,3-trans-24 and 2,3-cis-24……………26 2.3.4 Amide bond formation of scaffolds 2 and 3……………………………27 2.3.5 Preliminary assay for α-L-Iduronidase inhibitory activity…………………28 2.4 Conclusion…………………………………………………………………………29 Chapter 3. Experimental section………………………………………………………30 3.1 General experimental procedure…………………………………………………30 3.2 Procedure and experimental data of substrate 1…………………30 3.2.1 Standard route…………………………………………………………30 3.2.2 Modified route……………………………………………………………39 3.2.3 Azafuranose…………………………………………………………………43 3.3 Procedure of bioassay……………………………………………………………56 References…………………………………………………………………………………57 Appendix……………………………………………………………………………………614890436 bytesapplication/pdf論文使用權限：不同意授權亞胺醣環硝艾杜糖己醛醣酸鹽水解酵素抑制劑thesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/261260/1/ntu-102-R99223175-1.pdf