本論文為合成一系列可專一性標記醣蛋白的亞硼酸-苯基磺醯基化學探針分子，其組成包含與醣蛋白之醣體有親和力的配體官能基、具反應活性的苯基磺醯基反應中心以及做為報導官能基基團的末端炔。配體官能基是親和力導向基團，藉由與醣蛋白之醣基測鏈的雙醇結構形成酯基，將探針分子拉靠近醣蛋白表面，表面具親核性的胺基酸殘基能與反應中心進行親核性取代反應，將報導官能基成功藉由鄰近效應連結至醣蛋白表面上。本文中改變了配體官能基的結構以及苯基磺醯基反應中心的取代基，進而探討配體官能基的結構改變可否提升探針分子的標記效果以及改變苯基磺醯基取代基造成的電子誘導效應可否提升親核性取代反應的反應性，並且減少醣蛋白表面非專一性修飾的現象發生。

In this research, a series of boronic acid (BA)-tosyl chemical probes have been designed and synthesized. The synthetic probes composed of a boronic acid derivative as an affinity ligand, a tosyl group as a reactive group and a terminal alkyne reporting group were synthesizd and used in the labeling of glycoproteins. Boronic acid (BA) is an affinity-based funtional ligand, which can form a boronate diester ring with the diol groups on glycans. Following the initial step, the tosyl group act as a reactive center that allows a substituted reaction to take place with the nucleophilic residues of the glycoproteins, and conjugates the alkyne reporting group onto the glycoprotein surface via proximity effects. In this research, the labeling efficiency and specificity of glycoproteins was found to be considerably improved by modifying the structures and electronic properties of the tosyl reactive center.

論文審定書……………………………………………………….............……………i
誌謝……………………………………………………….............…………...........…ii
中英文關鍵字………………………………………………………………...………iii
中文摘要…………………………………………………………...................………iv
英文摘要……………………………………………………………………....………v
目次……………………………………………………………………....……….......vi
圖目錄………………………………………………………………………..….....…ix
流程目錄……………………………………………………………………...........… x
表目錄………………………………………………………………………...…..…..xi
光譜目錄……………………………………………………………………......……xii
縮寫表…………………………………………………………..…………...…...…xvii
第 一 章 緒論………………………………………………………………………..1
第一節 研究背景動機………………..…………………………………………..1
1.1 醣蛋白質及醣蛋白質體學………………..………………...........................1
1.2 醣蛋白質體學之研究方法………………..………………...........................3
1.2.1 分離及標記醣蛋白…………......................................................……3
1.2.2 亞硼酸及其衍生物於醣基化分子辨識之應用..................................6
第二節 研究設計…………....……………………………………..................…10
2.1 探針設計之研究…………....………………………………….…..............12
第 二 章 實驗結果與討論…………………………………………………………16
第一節 化學探針之合成..................................................................................…16
2.1.1 短鏈一級胺化合物分子 (3) 之合成...............................................18
2.1.2 親和性配位基團 (Boroxole) 化合物分子 (8) 之合成..................18
2.1.3 親和性配位基團 (Wulff-type) 化合物分子 (12) 之合成.............20
2.1.4 中鏈-苯基磺醯基化合物分子 (18) 之合成....................................21
2.1.5 BA-m-tosyl (19) 之合成.................................................................22
2.1.6 BX-m-tosyl (20) 之合成.................................................................23
2.1.7 中鏈-甲基-苯基磺醯基化合物分子 (26) 之合成...........................23
2.1.8 BA-m-tosyl-M (27) 之合成............................................................24
2.1.9 BX-m-tosyl-M (28) 之合成............................................................25
2.1.10 中鏈-甲基-氯-苯基磺醯基化合物分子 (34) 之合成...................25
2.1.11 BA-m-tosyl-MCl (35) 之合成......................................................27
2.1.12 BX-m-tosyl-MCl (36) 之合成......................................................27
2.1.13 中鏈-碘-苯基磺醯基化合物分子 (42) 之合成.............................28
2.1.14 BA-m-tosyl-I (43) 之合成.............................................................29
2.1.15 BX-m-tosyl-I (44) 之合成.............................................................30
第二節 化學探針分子引導之醣蛋白專一性標記實驗........................................31
2.2.1 亞硼酸-苯基磺醯基化學探針分子標定醣蛋白的標記測試..........31
2.2.2 亞硼酸-苯基磺醯基化學探針分子於複雜環境標定醣蛋白的效率比較………………………………………........................................33
2.2.3 Boroxole化學探針分子於單純系統及複雜環境標定醣蛋白測試………………………………………............................................36
第 三 章 結論…………………………………………………………………....…39
第 四 章 引用文獻……………………………………………………………....…40
第 五 章 實驗步驟與光譜數據………………………………………………....…44
第一節 儀器設備與藥品材料……………………………………………....…44
第二節 合成步驟與數據………………………………………………………46
第三節 醣蛋白標記、定量、一價銅催化環化反應與西方墨點法分析……80
5.3.1 化學探針標記醣蛋白之方法…………..………………............…..80
5.3.2 標定後醣蛋白之定量…………………………………............…....80
5.3.3 醣蛋白之一價銅催化環化反應……………………...................….81
5.3.4 西方墨點法於被標定醣蛋白之分析………………..........………..81
第 六 章 光譜資料…………………………………………………………………83

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