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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 王宗興(Tsung-Shing Wang) | |
dc.contributor.author | Po-Hsun Chen | en |
dc.contributor.author | 陳柏勳 | zh_TW |
dc.date.accessioned | 2021-06-08T00:47:00Z | - |
dc.date.copyright | 2020-08-21 | |
dc.date.issued | 2020 | |
dc.date.submitted | 2020-08-13 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17955 | - |
dc.description.abstract | 標靶型共價抑制劑可藉由與目標生物分子形成共價鍵而使其失活,而Alpha-鹵代酮是其中一類常用的反應基團。標靶型共價抑制劑在臨床和生醫研究上皆扮演著重要的角色,但由於其較高的化學反應性,脫靶效應和其導致的生物毒性是一大為人詬病之處。 可控釋放可能是這個問題的解答。藉由可控的侷限基團屏蔽其原先的反應性,使活性基團必須在特定條件下才能被釋出。同時,由於增強型螢光(Turn on fluorescence)在分子探針上的優勢,我們希望進一步衍生原本的侷限基團為一雙官能化的侷限基團,使其能同時屏蔽分子探針的螢光與反應性。 我們先利用三種簡單的模型分子驗證可控釋放和屏蔽反應性的可行性,並將光觸發的侷限基團應用於實際的模型標靶型共價抑制劑上。我們以全合成得到FMK及其類似物CMK,並將其接上侷限基團。我們以HPLC色譜圖證實了CMK可以藉由UV照射成功被釋放出來;同時,西方墨點法也證實侷限基團能屏蔽抑制劑對RSK2蛋白自磷酸化的抑制效果。雙官能化侷限基團的合成仍在進行且即將完成,我們希望能藉此探針即時顯影細胞中RSK2的分布情形。 | zh_TW |
dc.description.abstract | Targeted covalent inhibitors (TCIs) are small molecules designed to inactivate their biological targets by covalent binding, and alpha-haloketone is one of the electrophilic warheads commonly used. For years, TCIs have played important roles in both clinical trials and biological researches. However, due to intrinsic reactivity of the electrophilic warheads, off-target effect is one of the main concerns for TCIs, which results in cytotoxicity. Controlled releasing could be a strategy to overcome this problem. By masking reactivity with a triggerable protecting group, the warhead can be regenerated only under specific condition. Furthermore, for turn-on fluorescence is another powerful tool in molecular probes, we also aim to derive the original capping group into a dual-functional version, masking both fluorescence and reactivity. We first confirmed the viability of controlled releasing and reactivity attenuation by model alpha-haloketones. The light-responsive capping group was further applied to the real TCI model. FMK and its analog CMK was total synthesized and capped. HPLC chromatograph showed successful releasing of the original inhibitor CMK under UV irradiation, and we also confirmed the attenuation of inhibition reactivity toward autophosphorylation of RSK2 on Ser386 by Western blot analysis. Synthesis of the dual-functional trigger is on-going, and we hope to utilize this probe for real-time imaging of endogenous RSK2. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T00:47:00Z (GMT). No. of bitstreams: 1 U0001-1308202020575400.pdf: 11510368 bytes, checksum: d38d459a84609928b757ae694cadba17 (MD5) Previous issue date: 2020 | en |
dc.description.tableofcontents | 口試委員會審定書 # 誌謝 i 中文摘要 ii ABSTRACT iii CONTENTS iv LIST OF FIGURES vii Abbreviation x Chapter 1 Introduction 1 1.1 Targeted Covalent Inhibitors (TCIs) 1 1.1.1 The Advantages of Small Molecule Inhibitors 1 1.1.2 Introduction to TCIs 2 1.1.3 Common Warheads on TCIs 3 1.1.4 Alpha-haloketones 4 1.1.5 FMK and CMK, Inhibitors of RSK2 7 1.2 Controlled Releasing of Small Molecules 8 1.2.1 The Importance of Spatiotemporal Resolution 8 1.2.2 Inspiration from Prodrugs 9 1.2.3 Photocages 11 1.3 Small Molecules as Probes 12 1.3.1 Small Molecule Inhibitors as Probes 12 1.3.2 Turn-on Reactivity and Fluorescence 14 Chapter 2 Results and Discussions 17 2.1 Molecular Design 17 2.2 Synthesis of Models with Different Triggerable Capping Groups 19 2.2.1 Synthesis of 1,3-Dioxolane-capped Models 19 2.2.2 Synthesis of Oxazolidine-capped Models 21 2.3 Releasing Assay and Reactivity Assay 22 2.3.1 Releasing Assay of the ortho Nitro 1,3-Dioxolane-capped Models 22 2.3.2 Releasing Assay of the para Nitro 1,3-Dioxolane-capped Models 23 2.3.3 Releasing Assay of the Oxazolidine-capped Model 24 2.3.4 Reactivity Assay 25 2.4 Synthesis of Capped Model TCI 26 2.4.1 Total Synthesis of FMK and CMK 26 2.4.2 Synthesis of Capped FMK and CMK 28 2.4.3 Releasing of Dioxo-CMK 30 2.5 Inhibitor Activity Assay 30 2.5.1 Kinase Inhibition Assay 30 2.5.2 Cell Viability Assay 31 2.6 Synthesis of the Probe with Light-activatable Fluorescence and Reactivity 32 2.6.1 Synthesis of the Dual-functional Capping Group 32 Chapter 3 Conclusion and Perspective 35 Chapter 4 Material and Methods 36 4.1 General Synthetic Methods 36 4.2 Synthesis and Characterization of Compounds 37 4.2.1 Synthesis of Model Compounds 37 4.2.2 Synthesis of Model TCI FMK and CMK 47 4.3 Releasing Assay and Reactivity Attenuation Assay 63 4.3.1 General methods 63 4.3.2 Photo Releasing Assay 64 4.3.3 Enzymatic Releasing Assay 64 4.3.4 Reactivity Assay 64 4.4 Biological Assays 65 4.4.1 General Biological Methods 65 4.4.2 Cell Culture 66 4.4.3 Kinase Inhibition Assay 66 4.4.4 Western Blot 67 4.4.5 Cell Viability Assay 67 REFERENCES 69 | |
dc.language.iso | en | |
dc.title | 衍生Alpha-鹵代酮之可觸發侷限基團並應用於光調控螢光與反應性 | zh_TW |
dc.title | Deriving a Triggerable Capping Group for Alpha-haloketones and its Application in Light-activatable Fluorescence and Reactivity | en |
dc.type | Thesis | |
dc.date.schoolyear | 108-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 謝俊結(Jiun-Jie Shie),朱忠瀚(Chung-Han Chu) | |
dc.subject.keyword | 標靶型共價抑制劑,Alpha-鹵代酮,可控釋放,侷限基團,螢光探針, | zh_TW |
dc.subject.keyword | Targeted covalent inhibitors,Alpha-haloketones,Controlled releasing,Capping group,Fluorescent probes, | en |
dc.relation.page | 119 | |
dc.identifier.doi | 10.6342/NTU202003324 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2020-08-14 | |
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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