本研究旨在設計與合成可藉由β-葡萄醣醛酸酶(β-glucuronidase, βG)調控(off/on) 之近紅外光探針Glu-NIRoff-Quencher。其設計策略係將葡萄醣醛酸基通過一可自分解(self-immolative) 的芐基苯環中間棒，連接近紅外光發光基團和淬熄劑基團，使兩者之間有足夠近的距離，透過如π-相互作用、凡得瓦接觸…等形成分子內複合物，因為靜態和動態交互作用實現螢光猝熄。此探針，可被βG專一性水解，然後經由1, 6-脫去反應(1, 6-elimination reaction)釋放出螢光團或猝熄團，二者距離變大而交互作用消失，螢光恢復。可調控螢光探針Glu-NIRoff-Quencher具有背景低，專一性激活和非侵入性深層組織成像的優點，可以即時觀察某些腫瘤和腸道中βG的活性，將是加速癌症預防與治療的重要工具。通過相同的策略，我們也設計了Glu-NIRoff-Drug，是可藉βG調控的治療型探針。我們將喜樹鹼類化學治療藥物做為猝熄分子將近紅外光淬熄，因為是前驅藥，毒性較原形藥弱，經βG活化後毒性和螢光才恢復，達到同時殺死腫瘤細胞和顯影。Glu-NIRoff-Drug結合診斷和治療的功能，可以選擇性地靶向癌細胞，提供細胞毒性藥物，又同時在腫瘤組織中產生易於監測的成像信號，這有助於減少副作用和實時監測藥效的潛力，從而促進癌症個性化醫療的發展。

This study aims to design and synthesize a βG-based (off/on) near-infrared (NIR) probe, Glu-NIRoff-Quencher. In which glucuronic acid is connected via a self-immolative aromatic linker to NIR fluorophore and quencher. The control of intramolecular complex formation between fluorophore and quencher like π-interactions, van der Waals contact, etc., results in fluorescence quenching via both static and dynamic quenching. Upon hydrolyzed by βG specificity to liberate fluorophore or quencher through a 1, 6-elimination reaction, then restore the fluorescence. Fluorescence switchable Glu-NIRoff-Quencher probe has the advantages of low background, specific activation, and noninvasive deep-tissue imaging. It is a valuable tool for instantly visualizing βG activity in some tumors and intestinal to accelerate the development of personalized cancer treatment and prevention. By use of the same strategy, we design Glu-NIRoff-Drug as a βG-specific fluorescent theragnostic prodrug containing glucuronic acid, a chemotherapeutic drug, and NIR fluorophore along with an aromatic self-immolative linker. Chemotherapeutic drug plays the role of quenching and be released to kill tumor cells after activation. Therefore, Glu-NIRoff-Drug can target cancer cells selectively, offer a cytotoxic drug and produce readily monitored imaging signals in tumor tissues. It is foreseeable that combining specialized diagnosis and therapy would be the potential to enhanced anticancer efficacy, correct diagnosis and real-time monitoring for advancing the fields of personalized medicine.

目錄
中文摘要 I
英文摘要 II
致謝 III
目錄 IV
圖目錄 V
第一章 緒論 1
第二章 結果與討論 19
第一節 合成連接金奈米粒子的Glu-NIRoff-SH 19
第二節 合成探針Glu-NIRoff-Dabcyl 26
第三節 合成探針Glu-NIRoff-BQC 30
第三章 結論 34
第四章 實驗部分 37
第一節 儀器 37
第二節 試藥來源 38
第三節 合成方法 40
第五章 參考文獻 49
附件一、化合物光譜 55

圖目錄
圖 一、分子影像探針，體內顯示藥物治療靶點的分子改變 6
圖 二、螢光探針成像具高靈敏度、無損快速分析和實時檢測功能 6
圖 三、螢光分子探針“可激活(activatable)”或“可調控(off/on) 的概念” 7
圖 四、常見淬熄機制。PET和靜態淬熄需要螢光團和淬熄團碰觸 7
圖 五、依照不同螢光團選擇光譜重疊的暗淬熄團，經FRET淬熄 7
圖 六、偵測不同分析物的探針及其應用的淬熄原理 8
圖 七、分子內異(同)二聚體可調控探針的例子及淬熄策略 8
圖 八、酶受質螢光探針專一性活化的設計：(a )將螢光團分子接上可被酶識別的受質基團；(b)自消融中間棒連接識別基團和螢光團 12
圖 九、腫瘤細胞周圍有大量βG表現，可作為腫瘤標記設計前藥 12
圖 十、發展腸道βG抑制劑，減少βG抗癌前藥的代謝副作用 12
圖 十一、市面上有的β-葡萄醣醛酸酶活性檢測呈色劑和探針 14
圖 十二、FITC-TrapG和FITC-TrapG捕捉型探針的設計策略和結構 15
圖 十三、以FITC-TrapG偵測活體內腫瘤中β-葡萄醣醛酸酶的活性 15
圖 十四、以NIR-TrapG偵測活體內腫瘤中β-葡萄醣醛酸酶的活性 15
圖 十五、可調控βG近紅外光探針Glu-NIRoff-Quencher的策略和目標 18
圖 十六、可調控近紅外光治療型探針Glu-NIRoff-Drug的策略和目標 18
圖 十七、IR775、Dabcyl和化合物19在甲醇和水溶液中的吸收光譜 28

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