利用一鍋化策略製備硫酸乙醯肝素寡醣分子庫

Preparation of Heparan Sulfate Oligosaccharide Library Using One-pot Strategies

鍾永清(Yong-Qing Zhong)

指導教授 ： 廖文峯 ； 洪上程

國立清華大學/理學院/化學系/博士(2013年)

摘要

本論文主要論述細胞表面的硫酸乙醯肝素寡醣分子庫之合成研究，其一是以纖維細胞成長因子一型為作用目標，進而合成12種硫酸乙醯肝素的雙醣分子；其二為利用一鍋化醣鏈結反應建構4種可能的六醣結構，再進而合成出4個硫酸乙醯肝素。
第一章主要在介紹細胞表面的醣共軛分子，並說明硫酸乙醯肝素的結構及生物活性；第二章則是整理現行醣化學合成界中，主要的幾種醣鏈合成方法，包括傳統方式以及一鍋化製備的部分；第三章是在回顧探討關於與FGF結合、分子庫合成、及與單純疱疹病毒作用之硫酸乙醯肝素八醣體的合成相關文獻；第四章則敘述12種硫酸乙醯肝素雙醣分子的反合成分析。
第五章為合成12種硫酸乙醯肝素雙醣所需的單醣部份，特別是在L式艾杜醣衍生物的製備上，採取了不同的溶劑和反應溫度，使其產率大幅地上升；第六章則是將上述的兩個雙醣體進行偶合後，經數步的官能機轉換，得到共同中間體279，並以此化合物為基礎，經由變換去保護及官能基轉換反應的順序，即可獲得12種硫酸乙醯肝素雙醣分子；第七章主要介紹12種硫酸乙醯肝素雙醣分子的生化測試部分，在恆溫滴定測焓儀和蛋白質結晶的幫助下，更加了解硫酸乙醯肝素與FGF1的作用關係。
第八章是講述硫酸乙醯肝素寡醣於合成上的困難，並介紹為何引入一鍋化醣鏈結方式以及新式相對反應數值量測來製備醣鏈的想法；第九章則是敘述如何測量新式相對反應數值和所需的單醣體之製備，並試圖就所量測出的結果說明不同離去基的反應情況；第十章是敘述4種硫酸乙醯肝素六醣分子的反合成分析；第十一章主要介紹如何利用新式相對反應數值之結果，設計出一鍋化醣鏈結反應以合成出4種六醣骨架；第十二章則是選擇1種已合成出的六醣骨架，經幾步官能基轉換，製備出4種硫酸乙醯肝素六醣體。
在第十三章中，統整了第五章、第六章、第七章、第十一章及第十二章的合成工作和生化結果。第十四章則提供了合成工作的詳細實驗步驟及化合物的物理性質。

關鍵字

硫酸乙醯肝素 ； 一鍋化策略

並列摘要

This dissertation describes the synthesis of twelve heparan sulfate disaccharides for FGF-1 binding assay and the preparation of four hexasaccharides by one-pot strategy.
Chapter 1 introduces the cell surface glycoconjugates and explains the biological function of heparan sulfate. The literature reports for the preparation of oligosaccharide skeletons by stepwise methods or one-pot glycosylation and the synthesis of heparan sulfates that bind with FGF-1 and other proteins are summarized in Chapter 2 and Chapter 3, espectively. Chapter 4 describes the retro-synthetic plan for the twelve heparan sulfate disaccharides.
Chapter 5 describes the synthesis of two monosaccharides for generating twelve heparan sulfate disaccharides, including the optimization of 1,6-anhydro-L-idopyranose preparation. In Chapter 6, glycosylation of two prepared monosaccharides was carried out to yield the disaccharide skeleton followed by several functional group transformations to produce the common intermediate 279. The final twelve heparan sulfate disaccharides were synthesized by the subsequent deprotection and sulfonation strategies. Chapter 7 introduces two binding tests between heparan sulfate disaccharides and FGF-1 by ITC and protein crystallization. The information helps us understand the relationship between disaccharides and FGF-1 more than before.
Chapter 8 describes the difficulty in synthesizing heparan sulfate oligosaccharides and the reason of using one-pot glycosylation and new relative reactivity value to prepare oligosaccharide skeletons. In Chapter 9, various monosaccharides for measuring new relative reactivity values are synthesized. Based on these values, the relationship between reactivity of glycosyl thiol and leaving group is explained. Chapter 10 describes the retro-synthetic plan for four heparan sulfate hexasaccharides. Chapter 11 relates with the synthesis of four heparan sulfate hexasaccharide skeletons. The preparation of four final hexasaccharides is summarized in Chapter 12.
The conclusion of the present synthetic work is summarized in Chapter 13, a total synthesis of twelve heparan sulfate disaccharides and four hexasaccharides. Chapter 14 provides the detailed experimental procedure and physical data of the compounds synthesized herein.

並列關鍵字

Heparan Sulfate ； One-pot Strategies

參考文獻

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