本論文為利用本實驗室近期開發之新型螢光分子二氫喹啉-4-亞胺 (dihydroquinolin-4-imine)，與先前開發之蛋白質標記探針結合，將二氫喹啉-4-亞胺之兩起始物，N-丙炔苯胺 (N-propargylaniline) 與磺酰基疊氮化物 (sulfonyl azide) 分別裝上兩蛋白質探針並標記上目標蛋白，當兩目標蛋白接近並發生交互作用時，由於此兩探針上之起始物也同時接近並發生反應，生成二氫喹啉-4-亞胺產生螢光訊號，可建立發展出一套能觀察蛋白質間交互作用之光學觀測平台。而為了使二氫喹啉-4-亞胺的生成發生在蛋白質交互作用時，需先讓此反應能在水相中進行，本文中針對N-丙炔苯胺 (N-propargylaniline) 與磺酰基疊氮化物 (sulfonyl azide) 此兩起始物進行一系列水溶性改良，並對其結構修飾上帶電荷之四級胺官能基，使兩起始物分子擁有良好的水溶性，進而測試二氫喹啉-4-亞胺在水相生成之反應性。

In this research we have developed Cu (Ⅰ) catalysis synthesis of dihydroquinolin-4-imines via ketenimine intermediates. These dihydroquinolin-4-imine molecules have fluorescence properties and show very good quantum yields. Their environment-effect of the fluorescent properties can be used for the application of protein labeling. Furthermore, the strategy of this protein fluorescent labeling has the potential for developing the imaging technique of protein-protein interaction. In this research, we have attempted to synthesize dihydroquinolin-4-imine in aquesous phase which is the most important part and challenge for protein labeling. A series of water-solubility modification for these reactants of N-propargylaniline and sulfonyl azide has been designed and synthesized. Finally, these reactants modified with quaternary amine group to have good water-solubility, and then tested the reactivity for preparing the dihydroquinolin-4-imines in aqueous phase.

中文摘要…………………………………………………………...................……….v
英文摘要……………………………………………………………………....……...vi
目次……………………………………………………………………....………......vii
圖目錄………………………………………………………………………………....x
流程目錄……………………………………………………………………………...xi
表目錄……………………………………………………………………………….xiii
光譜目錄………………………………………………………………………….…xiv
縮寫表………………………………………………………………………………..xx
第一章 緒論…………………………………………………………………………..1
第一節 研究背景………………………………………………………………..1
1.1. 1. 三唑 (triazole) 雜環化學………………………………………........…..1
1.1.1.1. 雜環化學介紹………………………………………….............……..1
1.1.1.2. 1,2,3-三唑 (1,2,3-triazole) 的發展…………………………...............2
1.1.1.3. 一價銅催化疊氮與炔類環化加成 (CuAAC) 方法學探討...............4
1.1.1.4. 一價銅催化疊氮與炔類環化加成 (CuAAC) 方法學應用衍
生……………………………………………………………..6
1.1.2. 小分子螢光化學…………………………………………………….9
1.1.2.1. 螢光雜環分子之介紹………………………………………..9
1.1.2.2. 分子冷光之發光機制………………………………………10
1.1.2.3. 常見的螢光雜環分子………………………………………13
1.1.3. 螢光分子之蛋白質標記應用……………………………………...16
1.1.3.1. 螢光雜環分子與生物標記之發展…………………………16
1.1.3.2. 開關型螢光雜環分子的Bio-orthogonal labeling標記之介紹
………………………………………………………………21
1.1.3.3. 開關式雜環螢光分子蛋白標記技術的長期發展…………29
第二節 研究動機與實驗設計…………………………………………………30
1.2.1. 本實驗室開發一價銅催化疊氮與炔類環化加成之方法學……...30
1.2.1.1. 利用乙烯亞胺之中間體合成β胺基酸衍生物……………30
1.2.1.2. 利用銠金屬催化合成3-甲酰吲哚衍生物與苯并呋喃之合成
方法學………………………………………………………31
1.2.1.3. 加熱生成乙烯亞胺建構二氫喹啉-4-亞胺與chroman-4-
imine之方法學……………………………………………..35
1.2.1.4. 利用一價銅催化合成出具有螢光特性之二氫喹啉-4-亞胺
分子…………………………………………………………40
1.2.2. 研究動機及實驗設計……………………………………………...44
1.2.2.1. 研究動機與實驗目的………………………………………44
1.2.2.2. 實驗與模型之設計…………………………………………47
第二章 實驗結果與討論……………………………………………………………50
第一節N-propargylaniline基團片段水溶性修飾與水相反應測試………….50
2.1.1. 小分子N-propargylaniline一號模型實驗合成與結果討論……..50
2.1.1.1. N-propargylaniline一號模型設計與合成………………….50
2.1.1.2. N-propargylaniline一號模型水相反應測試……………….54
2.1.2. 小分子N-propargylaniline二號模型實驗合成與結果討論……..59
2.1.2.1. N-propargylaniline二號模型設計與合成………………….59
2.1.2.2. N-propargylaniline二號模型水相反應測試……………….62
2.1.3. 小分子N-propargylaniline三號模型實驗合成與結果討論……..63
2.1.3.1. N-propargylaniline三號模型設計與合成………………….63
2.1.3.2. N-propargylaniline三號模型水相反應測試……………….68
第二節 改善小分子模型水溶性與在緩衝溶液中之反應性測試……………69
2.2.1. 小分子N-propargylaniline三號模型之水溶性改善……………..69
2.2.1.1. 四級胺鹽類之N-propargylaniline分子設計與合成……...69
2.2.1.2. 四級胺鹽類之N-propargylaniline分子水相反應測試…...74
2.2.2. 小分子模型在緩衝溶液環境其反應性之測試…………………...75
2.2.2.1. 四號N-propargylaniline小分子模型在緩衝溶液之反應性
測試…………………………………………………………75
2.2.2.2. 磺酰基疊氮化物之分子模型設計合成與反應性測試……77
第三章 結論…………………………………………………………………………81
第四章 參考資料……………………………………………………………………82
第五章 實驗步驟與光譜數據………………………………………………………85
第一節 儀器設備與藥品材料…………………………………………………85
第二節 合成步驟與光譜數據…………………………………………………87
第六章 光譜資料…………………………………………………………………..129

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