ﰑﰎﰙﰚﰛﰜﰝﰞﰟﰠﰡﰢﰣﰤﰑﰥﰦﰧﰨﰑﰎﰙﰚﰛﰜﰝﰞﰟﰠﰡﰢﰣﰤﰑﰥﰦﰧﰨﰩ化學探針的設計與合成及其在生化方面之應用

Design, synthesis and evaluation of chemical probes for biochemical applications

許玉羚(Yu-Ling Hsu)

指導教授 ： 羅禮強

國立臺灣大學/理學院/化學研究所/博士(2015年)

https://doi.org/10.6342/NTU.2015.10172

摘要

化學探針能選擇性地標示目標蛋白質，為近代蛋白質體學上其中一種強力不可或缺的工具。本論文合成和設計幾種重要目標蛋白的化學探針，並介紹其發展及在生化方面的應用。第一部分針對蛋白激酶的結構特性設計的二個探針分子庫，皆以平行合成的組合式化學建構：第一個探針分子庫具有5'對苯磺醯氟腺苷和生物素標籤；第二個分子庫以3'疊氮去氧胸苷和腺苷為架構，接上三種親電子基。主架構的疊氮結構使其為一天然標籤基團。二個探針分子庫針對蛋白激酶的標示效能也在本論文進行驗証和比較。
穿心蓮內酯為一多功能的抗發炎和抗癌藥物，但相關的抑制機制卻還未完全明暸，因此建構具螢光團的穿心蓮內酯探針有助於進行生理中抑制和抗癌機制的追蹤和研究。第二部分合成和設計天然物穿心蓮內酯的可穿透細胞螢光探針，第一個可穿透細胞穿心蓮內酯螢光探針的合成經驗也有助於未來此類探針或抑制劑的開發和改良。本論文也合成α-L-岩藻糖苷酶醌甲基生成機制活性探針，以鹼催化置換β變旋異構物成α結構的關鍵中間體，並進而合成帶有螢光團的可穿透細胞

關鍵字

分子探針 ； 蛋白質體學 ； 對苯磺醯氟腺苷 ； 穿心蓮內酯 ； α-L-岩藻糖苷酶 ； 醌甲基 ； 同位素探針

並列摘要

Development of small-molecular tools which selectively react with designated protein families has been found powerful in modern functional proteomics. In this dissertation, we evaluated two kinase probe libraries according to the structure and functions of protein kinases. The first library adopted a 5'-p-fluorosulfonylbenzoyl adenosine (5'-FSBA) skeleton and a biotin reporter. The second library explored a novel molecular framework by attaching three kind of electrophilic warheads on 3'-azido 2', 3'-dideoxy base (AZT and AZA) recognition unit. The azido group at 3' position is a native clickable tag without further modification. The labeling performances of these two probe libraries toward kinases were compared.
Despite the therapeutic importance of this multifunctional herbal compound Andrographolide, the inhibition and regulation mechanism are still ambiguious. Herein, we designed a novel andrographolide-based, cell permeable fluorescent probe for in vivo identification of target proteins participating in related cancer or diseases. This first-time synthesized Andrographolide-based, cell permeable fluorescent probe provide valuable information for the synthesis method in the future. In addition, cell-permeable activity-based probes for α-L-fucosidase were evaluated by the key base-promoted epimerization step. These two probes carrying mono or difluoromethylphenyl group would generate reactive quinone methide as a trapping device after hydrolysis. In the last part, quantitative isotope-coded azido tags were synthesized and applied to detect the level of S-nitrosylation proteome with drug treatment.

並列關鍵字

probe ； protein profiling ； andrographolide ； α-L-fucosidase ； quinone methide ； isotope-coded tag

參考文獻

Chapter 1

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