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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 方俊民(Jim-Min Fang) | |
dc.contributor.author | Ming-Tyng Yeh | en |
dc.contributor.author | 葉明婷 | zh_TW |
dc.date.accessioned | 2021-06-07T18:19:54Z | - |
dc.date.copyright | 2012-02-16 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2012-01-17 | |
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Two different thymidylate kinase gene homologues, including one that has catalytic activity, are encoded in the onion yellows phytoplasma genome. 36. Barltrop, J. A.; Owen, T. C.; Cory, A. H.; Cory, J. G. Bioorganic & Medicinal Chemistry Letters 1991, 1, 611-614. 5-(3-carboxymethoxyphenyl)-2-(4,5-dimethylthiazolyl)-3-(4-sulfophenyl)tetrazolium, inner salt (MTS) and related analogs of 3-(4,5-dimethylthiazolyl)-2,5-diphenyltetrazolium bromide (MTT) reducing to purple water-soluble formazans As cell-viability indicators. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/16542 | - |
dc.description.abstract | 在人體中,胸苷三磷酸(dTTP)的供給與生成對DNA的合成與修補來說佔有舉足輕重的地位。而在胸苷三磷酸的生成過程中,胸苷酸(dTMP)必須經由胸苷酸激酶(thymidylate kinase, TMPK)的磷酸化過程形成胸苷二磷酸(dTDP),再透過核苷酸激酶的磷酸化進而產生胸苷三磷酸。干擾胸苷三磷酸的生成使細胞修復機制受損,已經是許多癌症化學治療藥劑發展的重要策略。在張智芬教授與胡春美博士的研究中顯示,以小片段干擾RNA的方式抑制胸苷酸激酶的表現,可有效降低胸苷三磷酸的含量並促使癌細胞敏感於基因毒化物的毒殺能力。但由於小片段干擾RNA的應用仍然受限於其遞送的效率,因此利用小分子去抑制胸苷酸激酶的活性應是較可行的策略。
在本篇論文中,我根據胡春美博士高效率藥物篩選的結果,針對一有效的前驅人類胸苷酸激酶抑制劑(YMU1)建立合成路徑,並在張智芬教授實驗團隊的幫助下,對其抑制劑作細胞與活鼠實驗,證實其化合物有效抑制人類胸苷酸激酶,並增加低劑量癌症藥物小紅莓(Doxorubicin)所誘導的細胞死亡。為了瞭解YMU1對胸苷酸激酶的抑制活性,並找尋更好的抑制劑,我設計並合成一系列YMU1的衍伸物,測試其結構與活性之間的關係。根據我們的研究結果,目前已有13個YMU1衍伸物已被證實具有與YMU1相同程度的抑制能力,並依所有44個YMU1衍伸物的結構與抑制活性的關係,推論出針對人類胸苷酸激酶有效抑制的化學結構,以利未來抑制劑的開發。 | zh_TW |
dc.description.abstract | Human thymidylate kinase (hTMPK) plays an essential role in dTTP supplement by catalyzing the reversible phosphoryl transfer from a donor (preferably ATP) to the acceptor TMP. The process of dTTP biosynthesis is required for efficient DNA replication and repair in cells. Introduction of genotoxic insult has been a major chemotherapeutic dogma in cancer treatment; Hu and Chang have shown that blocking dTTP supply could amplify DNA damage signal in highly proliferating tumor cells, thereby sensitizing these cells to chemotherapeutic agent such as doxorubicin, a topoisomerase II inhibitor.
In my research, I aim to find cell permeable small molecules as to block hTMPK function. Based on the luciferase-coupled TMPK assay, a heterocyclic compound, YMU1, was found to be an hTMPK inhibitor. Many YMU1 derivatives were synthesized, and their effects on TMPK inhibition were evaluated. Moreover, inhibition of TMPK enhanced tumor lethality when treated with a low dose of chemotherapeutic agent. The combined treatment is a potential novel chemotherapy against cancer cells. The general structural features of YMU1 derivatives include (1) a heterocycle H1 that may comprise 1–4 heteroatoms of N and O elements, (2) 1–3 substituents, G, on H1, (3) another heterocycle H2 in the linker, and (4) a library of R groups. According to our study, we have synthesized and identified other 13 compounds having comparable level of inhibitory activity against hTMPK like YMU1. | en |
dc.description.provenance | Made available in DSpace on 2021-06-07T18:19:54Z (GMT). No. of bitstreams: 1 ntu-100-R98223168-1.pdf: 8624732 bytes, checksum: fd5c28180af9d8330762334a09677b6b (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | Abstract in Chinese I
Abstract in English II Table of Contents V Index of Figures VIII Index of Schemes XI Index of Tables XII Abbreviations XIII Chapter 1. Introduction 1 1.1 The importance of dTTP in cells 1 1.2 Biosynthetic mechanism of dTTP in human cell 2 1.3 Cancer treatments by dTTP deprivation 3 1.4 Cancer chemotherapeutic strategies of 5-FU 4 1.5 Scrutiny of thymidylate kinase (TMPK) 6 1.6 Physiology of TMPK 7 1.7 Chemotherapeutic value of TMPK 11 1.8 Inhibition of DNA topoisomerase II 14 1.9 Synthestic lethality 17 1.10 TMPK silence 19 1.11 High-throughput screening 21 Chapter 2. Results and Discussion 26 2.1 YMU1 synthesis 26 2.1.1 Retrosynthesis 26 2.1.2 Selectivity of N- or O-alkylations 28 a. Solvent effects 31 b. Effect of bases 32 c. Effect of leaving groups 34 2.2 Characterizing YMU1 as an hTMPK inhibitor 38 2.2.1 Inhibitory activity of YMU1 39 2.1.2 In vitro and in vivo chemosensitization by YMU1 treatment 42 2.3 Construction of new pharmacophore models 46 2.3.1 1st pharmacophore model 46 2.3.1.1 General synthetic scheme 47 2.3.1.2 Structure–activity relationship 50 2.3.2 2nd pharmacophore model 56 2.3.2.1 General synthetic scheme 58 2.3.2.2 Structure–activity relationship 60 2.4 Conclusion 65 Chapter 3. Experimental Section 67 3.1 General part 67 3.2 Determination of the purity of compounds 68 3.3 Expressin and purification of enzymes 68 3.4 Luciferase-coupled TMPK assay 68 3.5 NADH-coupled TMPK assay 69 3.6 Cell treatment and MTS cytoxicity assay 69 3.7 In vivo chemotherapy 70 3.8 Synthetic procedures and characterization of compounds 71 References 105 Appendix-1. 1H and 13C NMR spectra 112 Appendix-2. HPLC diagrams 185 | |
dc.language.iso | en | |
dc.title | 人類胸苷酸激酶抑制劑的合成與鑑定 | zh_TW |
dc.title | Synthesis and Identification of Novel Chemosensitizers against Human Thymidylate Kinase | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-1 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 張智芬(Zee-Fen Chang) | |
dc.contributor.oralexamcommittee | 陳平(Richard P. Cheng),許世宜(Sheh-Yi Sheu) | |
dc.subject.keyword | 胸苷,酸激酶,抑制劑, | zh_TW |
dc.subject.keyword | Chemosensitizers,Human Thymidylate Kinase, | en |
dc.relation.page | 189 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2012-01-18 | |
dc.contributor.author-college | 理學院 | zh_TW |
dc.contributor.author-dept | 化學研究所 | zh_TW |
顯示於系所單位: | 化學系 |
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