臺灣大學: 化學研究所陳昭岑胡宗豪Hu, Tsung-HaoTsung-HaoHu2013-04-112018-07-102013-04-112018-07-102010http://ntur.lib.ntu.edu.tw//handle/246246/257356在細胞分裂期間，端粒長度的縮減可能會造成基因組的不穩定以及細胞衰老，但是在大部分的腫瘤細胞當中有著過度表現的端粒酶，藉著不斷地增加端粒的長度來達到無限制的增殖。而令我們有興趣的是當端粒摺疊成鳥嘌呤四股結構(G-quadruplex)時，是無法被端粒酶所延長的。近年來咔唑衍生物 BMVC 在文獻中被報導具有穩定鳥嘌呤四股結構的潛力，並如預期地可以抑制端粒酶的活性，並且與鳥嘌呤四股結構嵌合時有螢光增強的效果。根據文獻中的理論模擬計算結果，BMVC 與鳥嘌呤四方結構 (G-quartet) 堆疊的位置在鳥嘌呤四股結構的上方或下方。 在此我們引入了氮芥子氣 (N-mustard) 來當作 DNA 烷化劑並結合 BMVC 的端粒辨識能力，以期能與堆疊位置附近的 DNA 鹼基形成共價鍵結，成為更強的鳥嘌呤四股結構穩定劑。因此在本論文中設計了不同長度的連接單元，用以連接 BMVC 及氮芥子氣，以期能藉由 BMVC 與四股結構結合之時，進而與空間中靠近的 DNA 鹼基進行烷化反應，並探討不同長度的連接單元是否影響四股結構辨識能力及 DNA 烷化能力。 這些合成出來的 BMVC 衍生物不但具有螢光性質，也有強的化學接合能力。我們藉由吸收、螢光、圓二色光譜以及凝膠電泳實驗等方法，來分析 BMVC 衍生物與鳥嘌呤四股結構的作用能力，另一方面以質譜來分析與 DNA 的共價鍵結產物。The reduction of telomeres length during cell division can lead to genomic instability and senescence.1 Telomere repeats are added by telomerase, which is overexpressed in most tumor cells for the unlimited cell proliferation. Of particular interest is that telomeres can fold into G-quartet structure which cannot be elongated by telomerase. Recently, 3,6-Bis(1-methyl-4-vinylpyridinium)carbazole diiodide (BMVC), a G-quartet stabilizer, was reported, showing telomerase inhibition ability along with fluorescence enhancement while intercalating with G-quadruplex.2 Based on the simulation results, BMVC intercalated with G-quartet on the top or bottom of the G-tetrad more likely.3 Consequently, introducing nitrogen mustard as DNA-alkylating agent tethered to BMVC could form covalent linkage with neighboring DNA backbone, rendering it become a stronger G-quartet stabilizer. In this thesis, we designed linkers with three different length, which links BMVC and nitrogen mustard, hoping to covalently linked with neighboring bases while binding with G-quadruplex. These BMVC derivatives possess high fluorescence quantum yield as well as show strong chemical ligation ability. We analyzed the interactions between BMVC derivatives and G-quadruplex by UV/Vis spectroscopy, fluorescence spectroscopy, circular dichroism, as well as DNA-PAGE studies. On the other hand, we successfully detected and analyzed DNA adducts by mass spectrometry.14940020 bytesapplication/pdfen-US端粒鳥嘌呤四股結構烷化劑端粒?抑制劑telomereG-quadruplexalkylating agentstelomeraseinhibitorthesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/257356/1/ntu-99-R97223167-1.pdf