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標題: | 瘤克寧類似物的設計與合成應用於硝基還原酶活化螢光顯影治療之前體藥物 Design and Synthesis of Chlorambucil Analogues for Nitroreductase-Activated Fluorescent Theranostic Prodrugs |
作者: | Tzu-Yu Yu 俞姿宇 |
指導教授: | 陳昭岑(Chao-Tsen Chen) |
關鍵字: | 苯胺氮芥,DNA烷化,缺氧腫瘤顯影,顯影兼治療型前驅藥,硝基還原?, Chlorambucil,DNA alkylation,hypoxic tumor imaging,theranostic prodrug,nitroreductase, |
出版年 : | 2018 |
學位: | 碩士 |
摘要: | 本論文的主要目的是設計和構建一系列新的DNA烷化劑,以提高DNA烷基化效率和現有烷化劑的選擇性。苯胺氮芥的反應性可以通過改變苯胺氮上取代基的電子性質來調節。
論文第一部分設計並合成了DNA烷基化劑COOH-SW和COOH-SW-F,其中給電子基(甲基)和拉電子基(三氟甲基)分別連接在苯胺的氮原子上,調節苯胺氮芥對DNA烷基化的反應性。通過高效能液相層析儀和電噴灑游離法質譜分析驗證了苯胺氮芥衍生物的反應性。研究得出水解速率排列為Chlorambucil ≥ COOH-SW > COOH-SW-F,表明氮上的電子密度越大,水解速率越快。 論文第二部分則是設計並合成缺氧腫瘤的顯影兼治療型前驅藥物AO-NMB和BT-NMB。將DNA辨識基團吖啶和苯並噻唑修飾在分子上,導引AO-NMB和BT-NMB接近DNA,使烷基化更容易。通過氨基甲酸酯基團將對-硝苄基與吖啶(AO)或苯並噻唑(BT)結合,以抑制烷基化活性並淬滅AO和BT的螢光,在低氧細胞中過表達的硝基還原酶或化學還原劑的反應下切除對-硝苄基。在加入模擬過量表達的硝基還原酶的化學還原劑後,AO-NMB發生還原,接著發生1,6-消除反應,並釋放具烷基化能力的螢光分子AO-NM。通過變性聚丙烯醯胺膠體電泳實驗分析AO-NM在雙鏈DNA上的烷基化效率。結果表明,與Chlorambucil相比,AO-NM顯示出更好的DNA烷基化活性。在BT-NMB的情況下,化學還原劑不能有效地將BT-NMB有效地還原為BT-NM。為了驗證BT-NM是否具有烷基化能力,BT-NM是獨立合成的,電泳實驗表明BT-NM可以使雙鏈DNA單烷基化,烷基化產率高達76%,而無DNA辨識基團的COOH-SW僅顯示50%單烷基化產率。 The main objective of this thesis is to design and construct a new series of DNA alkylating agents to improve DNA alkylation effectiveness and the selectivity of current existing alkylating agents. The reactivity of aniline N-mustard warhead could be regulated by varying the electronic nature of the substituents on the aniline nitrogen. In the first part of the thesis, DNA alkylators COOH-SW and COOH-SW-F were designed and synthesized, where the electron-donating (methyl group) and -withdrawing (trifluoromethyl group) are attached on the nitrogen atom of aniline N-mustard, respectively, to regulate the reactivity of aniline N-mustard toward DNA alkylation. The reactivity of aniline N-mustard derivatives was validated by the hydrolysis experiment by HPLC and electrospray ionization mass spectrometry analysis. The results suggested that hydrolysis rate ranked in the decreasing order is Chlorambucil ≥ COOH-SW > COOH-SW-F indicating that the electron density on the nitrogen effectively influences the hydrolysis rate: the more electron density the faster hydrolysis rate. In the second part of the thesis, AO-NMB and BT-NMB were designed and synthesized. They can serve as theranostic prodrugs for hypoxic tumors. The DNA binders such as acridine and benzothiazole were incorporated into the molecule designs to direct AO-NMB and BT-NMB close to the DNA greatly rendering the alkylation more feasible. Moreover, p-nitrobenzyl group which can respond to the overexpressed nitrogen reductase in the hypoxic cells or chemical reductants was incorporated to either acridine (AO) or benzothiazole (BT) via carbamate group to mask the alkylation activity as well as quench the fluorescence of AO and BT. Upon the addition of the chemical reducing agents mimicking the overexpressed nitrogen reductase to AO-NMB, the reduction followed by 1,6-elimination reaction occurs and the fluorescent molecule AO-NM accompanied with the increased alkylation ability was released. The alkylation efficiency of AO-NM on double-stranded DNA was further evaluated and analyzed by denaturing PAGE electrophoresis. The results indicated that AO-NM showed better DNA alkylation activities compared to Chlorambucil. In the case of BT-NMB, chemical reducing agent somehow can’t effectively reduce BT-NMB to BT-NM. To validate BT-NM whether it exhibits alkylation ability, BT-NM was synthesized independently and the electrophoresis experiments indicated BT-NM can monoalkylate double-stranded DNA with the alkylation yield up to 76% whereas COOH-SW lack of DNA binder only showed 50% monoalkylation yield. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77782 |
DOI: | 10.6342/NTU201801638 |
全文授權: | 有償授權 |
電子全文公開日期: | 2028-07-17 |
顯示於系所單位: | 化學系 |
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