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研究生： 徐子凡
Hsu, Zih-Fan
論文名稱： 探討雙高石膽酸衍生物對於唾液酸轉移酶及癌細胞轉移的影響
Bishomolithocholic Acid: A Unique Template for Human Sialyltransferase Inhibitor, Optimized to Suppress Cancer Metastasis
指導教授： 李文山
Li, Wen-Shan
林文偉
Lin, Wen-Wei
學位類別： 碩士
Master
系所名稱： 化學系
Department of Chemistry
論文出版年： 2019
畢業學年度： 107
語文別： 英文
論文頁數： 96
中文關鍵詞： 唾液酸 、唾液酸轉移酶 、癌症轉移 、雙高石膽酸
英文關鍵詞： sialic acid, sialyltransferase, metastasis, bishomolithocholic acid
DOI URL： http://doi.org/10.6345/NTNU201900320
論文種類： 學術論文
相關次數： 點閱：222  下載：0 
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    • 在台灣，癌症為十大死因之首，其中癌細胞轉移為九成癌症病人之死因。癌細胞表面具有過度表現的唾液酸是細胞癌化的重要指標，此現象不僅會促進癌細胞的生長，亦可增加其侵犯周邊組織與轉移之能力。已有實驗證實，藉由調控唾液酸轉移酶能影響細胞表面之唾液酸表現。因此，抑制唾液酸轉移酶的活性會是一個有潛力的癌症轉移治療方式。
    我們合成雙高石膽酸衍生物HZF01-04，並進行初步生物試驗的研究與探討。HZF01-04具有極高的選擇性唾液酸轉移酶抑制能力，針對ST6Gal I有良好的抑制效果，其IC50數值介於7.8-12.2 μM之間。此外，HZF01-04對於人類三陰性乳癌細胞株MDA-MB-231的轉移能力具有抑制效果，其IC50數值之範圍介於7.5-10 μM之間。進一步的生物活性測試正在進行中，包括物化性質與動物實驗。
    此研究結果有助於選擇性唾液酸轉移酶抑制劑的開發，希望能應用於N-醣鏈過度唾液酸化的癌症病人之治療。

    Cancer is the major cause of death and metastasis is the greatest contributor to cancer deaths. Tumor cells display elevated expression of sialic acid on membranes, which promote cancer cell metastasis and progression. Aberrant sialylation is correlated with poor patient prognosis. Therefore, interfering hypersialylation through inhibition of sialytransferase activity may be a therapeutic target to metastatic cancer.
    In this investigation, we synthesized a series of bishomolithocholic acid derivatives HZF01-04 with various length linkers. HZF01-04 demonstrated significant inhibitory selectivity toward N-glycan sialyltransferase, β-galactoside α-2,6-sialyltransferase (ST6Gal I). The selectivity ratio of N-glycan vs O-glycan is over 80-fold. The findings show that HZF01-04 suppress serum-induced cancer cell migration in a dose dependent manner. Further studies for their mechanism of action and animal model are in progress.
    These efforts pave the way for the development of selective sialyltransferase inhibitors as effective anti-metastatic agents for the treatment of patients with aberrant overexpression of N-glycan sialylation.

    中文摘要 I Abstract II 謝誌 III Contents IV Scheme Contents VII Table Contents VIII Figure Contents IX List of Abbreviation X Awards & Honors XII I. Introduction 1 1.1. Background 1 1.2. Sialic Acid 2 1.3. Sialyltransferase 4 1.4. Sialylated Antigen 7 1.4.1 Sialyl Lewis antigens (SLea and SLex) 7 1.4.2 Truncated O-glycans 8 1.4.3 Highly branched N-glycan 8 1.4.4 Polysialic acid (polySia) 8 1.4.5 Glycosphingolipid (GSL) 9 1.5. Sialylation in Cancer Metastasis 9 1.5.1 Metastatic process 9 1.5.2 Role of glycoconjugates in metastasis 10 1.6. Sialyltransferase Inhibitors 12 1.6.1 Donor analogous inhibitors 12 1.6.2 Transition-state analogous inhibitors 13 1.6.3 Acceptor analogous inhibitors 13 1.6.4 Other sialyltransferase inhibitors 14 1.7. Previous Study 15 1.8. Research Motivation 19 II. Results and Discussion 21 2.1. Synthesis of Bishomolithocholic Acid Derivatives HZF01-04 21 2.1.1. Retrosynthesis 21 2.1.2. Synthesis of PEG linker 1a-1d 22 2.1.3. Synthesis of bishomolithocholic diacid 10 23 2.1.4. Synthesis of bishomolithocholic acid derivatives 25 2.2. In Vitro Studies of HZF01-04 27 2.2.1. Cytotoxicity of HZF01-04 in MDA-MB-231 27 2.2.2. Inhibition of ST3Gal I and ST6Gal I 28 2.2.3. Antimigration activity of HZF01-04 29 2.2.4. Cytotoxicity of HZF01-04 in MDA-MB-231 (SFM) 32 2.3. Preliminary Structure-Activity Relationship Studies 33 III. Conclusions 37 IV. Material and Methods 39 4.1. General Experimental 39 4.2. Synthetic Procedures 40 4.2.1. Preparation of PEG linker 40 4.2.2. Preparation of bishomolithocholic acid 43 4.2.3. Preparation of HZF01 46 4.2.4. Preparation of HZF02 49 4.2.5. Preparation of HZF03 52 4.2.6. Preparation of HZF04 55 4.3. Cell Line Used and Cell Culture 58 4.4. MTT Cytotoxicity Analysis 58 4.5. Sialyltransferase Assay 58 4.6. Transwell Migration Assay 59 V. References 61 Spectra 69

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