臺灣博碩士論文加值系統

香豆素(Coumarin)及其衍生物存在於許多天然物中，除了可作為香料使用外，由於本身具有多樣的生物活性，例如抗菌、抗癌或抗凝血劑等等醫藥用途，是個相當重要的分子。而Coumarin結構本身具有高度的螢光特性，因此長期以來廣泛的被運用在多種染料上。文獻上有許多報導，當藉著修飾或置換上不同取代基，即可微調其螢光波長及強度。我們利用Sonogashira耦合反應將3-acetyl-2-oxo-2H-chromen-7-yl trifluoromethanesulfonate上 7號碳修飾成苯炔基取代，嘗試延伸其共軛性質，使其吸收及放射波長紅位移。實驗結果發現，藉由改變不同推拉電子取代基，其吸收及放射波長也會有所差異。並且我們將此系列Coumarin衍生物運用到人類子宮頸癌細胞株，測試其活性，當藉由照光後(365 nm，5 min)發現相較於不照光毒性差異達150倍左右，具有成為光動力治療潛力之藥物，並進一步探討其死亡途徑，更利用其螢光特性應用於生物影像系統。

Coumarin derivatives are a calss of nature compounds presented in many plants and most of then possess antifungal, antibacterial, anti-tumour, and antibiotic activites etc. The coumarin moiety as chromophore of fluorescent indicator is a useful physical character in imaging technique in biological systems. Our recent attempts in the modification of 7-OTf-coumarine with alkynes by the method of Sonogashira reaction led to the formation of several coumarin derivatives with expected red-shift of wavelength in fluorescence. Applications of these compounds in the study of cytotoxicity towards HeLa cells have been carried out. Without irradiation of light, all these compounds exhibited negligible cytotoxicity. However, upon irradiation with selected UV light, some of these compounds exhibit fair to good photocytotoxicity toward HeLa cells. Base on their unique fluorescence properties, application of coumarin derivatives in bio-imaging technique to locate the distribution of compounds in cell were pursued.

第一章 序論 1
第一節 耦合反應(Coupling Reactions) 簡介 2
第二節 香豆素(Coumarin) 簡介及應用 9
第三節 螢光原理 17
第四節 細胞週期(Cell Cycle) 20
第五節 細胞凋亡(Apoptosis)與細胞壞死(Necrosis) 23
第六節 活性氧化物種 (Reactive oxygen species，ROS) 26
第七節 粒腺體膜電位 (Mitochondrial membrane potential) 27
第八節 光動力療法 (Photodynamic therapy，PDT) 28
第九節 研究方向 30
第二章 結果與討論 31
第一節 化合物3-Acetyl-7-hydroxy-2H-chromen-2-one 的製備 32
第二節 化合物3-Acetyl-2-oxo-2H-chromen-7-yl trifluoromethanesulfonate 的製備 33
第三節 利用 Sonogashira 耦合反應合成 Coumarin 衍生物 34
第四節 Coumarin 衍生物之吸收及螢光光譜 39
第五節 Coumarin 衍生物對人類子宮頸癌細胞存活率的影響43
第六節 Coumarin 衍生物產生活性氧化物種(Reactive oxgen species， ROS)之探討 49
第七節 Coumarin 衍生物對粒腺體膜電位之影響 52
第八節 Coumarin 衍生物誘導細胞凋亡之探討 54
第九節 Coumarin 衍生物在細胞內螢光影像拍攝 57
第三章 結論 59
第四章 實驗部分 60
第一節 儀器、試劑和操作技巧 61
第二節 實驗製備與方法 68
第三節 產物鑑定 82
參考文獻 94
附錄 104
化合物光譜數據 105
化合物 P2 晶體數據 142
化合物 8 晶體數據 150

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