請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/97121完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王宗興 | zh_TW |
| dc.contributor.advisor | Tsung-Shing Wang | en |
| dc.contributor.author | 黃建嘉 | zh_TW |
| dc.contributor.author | Jian-Jia Huang | en |
| dc.date.accessioned | 2025-02-27T16:17:27Z | - |
| dc.date.available | 2025-02-28 | - |
| dc.date.copyright | 2025-02-27 | - |
| dc.date.issued | 2025 | - |
| dc.date.submitted | 2025-02-11 | - |
| dc.identifier.citation | (1) Muttenthaler, M.; King, G. F.; Adams, D. J.; Alewood, P. F. Trends in peptide drug discovery. Nat. Rev. Drug Discov. 2021, 20 (4), 309-325.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/97121 | - |
| dc.description.abstract | 肽醛類化合物因其能與目標蛋白形成可逆共價鍵，被廣泛用作高效的抑制劑。然而，其高反應性容易導致脫靶效應，而且代謝不穩定性導致快速降解，限制了其更廣泛的應用。為了解決這些問題，我們開發了以噁唑烷保護醛基的策略，該策略結合了多種刺激響應性觸發基團來降低醛基的反應性，並能透過特定條件來釋放出醛基。
我們以MG132和Ac-LVK-CHO兩個肽醛為基礎合成的前驅藥物在微酸性和生理條件下皆有很好的水解穩定性。酵素抑制實驗也證實，此保護策略顯著降低了肽醛的活性；且在特定條件下觸發去保護反應恢復了與原藥相當的酵素抑制活性。在正交性實驗顯示，MG132-2NB和Ac-LVK-DiCN表現出部分正交性而MG132-pinBB和Ac-LVK-NIm分別對透過H2O2和nitroreductase去保護，表現出幾乎完全的正交性，在western blot實驗也證明可調控特定藥物被去保護影響細胞內蛋白表達。 因此本研究證明了以噁唑烷保護醛基的策略可以調控肽醛的活性，並且可選擇性和可控性地讓特定肽醛恢復活性，有助於增加肽醛的專一性。 | zh_TW |
| dc.description.abstract | Peptide aldehydes are widely utilized as potent inhibitors due to their ability to form reversible covalent bonds with target proteins. However, their high reactivity leads to off-target effects, and their metabolic instability results in rapid degradation, posing significant challenges for their broader application. To address these issues, we developed an oxazolidine-based caging system that incorporates diverse stimuli-responsive trigger groups, enabling precise spatiotemporal control of inhibitor activation. The oxazolidine cage allowed the integration of trigger groups such as photolabile (2-nitrobenzyl and coumarin derivatives), ROS-sensitive (aryl pinacol boronate), and hypoxia-responsive (nitroimidazole) groups, offering flexibility in prodrug design. The synthesized prodrugs demonstrated excellent hydrolytic stability under mildly acidic (pH 5.0) and physiological (pH 7.4) conditions.
Enzyme inhibition assays confirmed that caging significantly reduced inhibitor activity, with IC50 values increasing up to 153-fold for MG132 derivatives and 92-fold for Ac-LVK-CHO derivatives. Controlled decaging using specific stimuli, such as UV light (365 nm), visible light (520 nm), reactive oxygen species (H2O2), or nitroreductase, restored enzymatic inhibition activity to level comparable to those of the uncaged parent drugs. Orthogonality experiments revealed that MG132-2NB and Ac-LVK-DiCN exhibit partial orthogonality. Photodecaging at 365 nm activated both MG132-2NB and Ac-LVK-DiCN, while 520 nm selectively triggered Ac-LVK-DiCN decaging. Similarly, MG132-pinBB and Ac-LVK-NIm responded selectively to H₂O₂ and nitroreductase, respectively, showing nearly complete orthogonality. This study establishes the oxazolidine-based caging system as a robust platform for improving the stability, selectivity, and controllability of peptide aldehyde inhibitors, providing a versatile solution to challenges in reactivity and specificity. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2025-02-27T16:17:27Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2025-02-27T16:17:27Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 致謝 i
摘要 ii Abstract iii Table of Contents v List of Figures vii Abbreviations x Chapter 1. Introduction 1 1.1. Peptide drug 1 1.2. The Characteristic of Aldehyde Warhead 2 1.3. The Strategy of Protected Aldehyde Group 5 Chapter 2 Results and Discussions 11 2.1 Molecular Design 11 2.2 Synthesis 14 2.3. Determination of Hydrolytic Stability 22 2.4. Enzymatic Inhibition Assays of MG132, Ac-LVK-CHO and Their Prodrugs 23 2.5. Validation of Decaging Efficiency for MG132 Prodrugs to Release Peptide Aldehyde Under Specific Conditions 26 2.6. Validation of Decaging and Peptide Aldehyde Release for Ac-LVK-CHO Prodrugs by LC-MS and HPLC 28 2.7. Enzymatic Activity Assays Demonstrating the Inhibitory Effects of Prodrugs Before and After Decaging 31 2.8 Cell Viability of MG132 and Their Derivatives 34 2.9. Cell Viability of Ac-LVK-CHO and Their Derivatives 36 2.10. Verification of Cellular Response to Prodrug Before and After Decaging via Western Blot 38 2.11. Orthogonality of Photo-responsive Trigger Demonstrated by HPLC Analysis 46 2.12. Orthogonality of ROS- and Hypoxia-Responsive Triggers Demonstrated by HPLC Analysis 49 2.13. Western Blot Validation of Selective Decaging: MG132-pinBB by H₂O₂ and Ac-LVK-NIm by Hypoxia 53 2.14. Orthogonality of ROS- and Hypoxia-Responsive Triggers Demonstrated by Western Blot 55 Chapter 3. Conclusion and Perspectives 58 Chapter 4. Materials and Methods 60 4.1. General Methods and Instruments 60 4.2. Cell Culture 61 4.3. Determination of Hydrolytic Sability 61 4.4. 20S Proteasome Enzyme Inhibition Assay 62 4.5. Cathepsin B Enzyme Inhibition Assay 63 4.6. Cell Viability Assay 65 4.7. Validation of Decaging Efficiency by HPLC 68 4.8. Western Blot Analysis 71 4.9. Enzymetic Assay of Prodrug Decaging 75 4.10. Synthesis and Characterization of Compounds 77 Referances 111 Appendix I 118 Appendix II: Supplementary 153 | - |
| dc.language.iso | en | - |
| dc.subject | 活性調控 | zh_TW |
| dc.subject | 肽醛 | zh_TW |
| dc.subject | 噁唑烷保護 | zh_TW |
| dc.subject | 正交性 | zh_TW |
| dc.subject | Peptidyl aldehyde | en |
| dc.subject | Orthogonality | en |
| dc.subject | Activity regulation | en |
| dc.subject | Oxazolidine cage | en |
| dc.title | 帶有多功能性與正交性觸發基團的噁唑烷籠型結構用於控制肽醛活性 | zh_TW |
| dc.title | Oxazolidine-Based Cages with Versatile and Orthogonal Triggers for Controlling Peptide Aldehyde Activity | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 113-1 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 朱忠瀚;謝俊結 | zh_TW |
| dc.contributor.oralexamcommittee | Chung-Han Chu;Jiun-Jie Shie | en |
| dc.subject.keyword | 肽醛,噁唑烷保護,活性調控,正交性, | zh_TW |
| dc.subject.keyword | Peptidyl aldehyde,Oxazolidine cage,Activity regulation,Orthogonality, | en |
| dc.relation.page | 153 | - |
| dc.identifier.doi | 10.6342/NTU202500593 | - |
| dc.rights.note | 同意授權(全球公開) | - |
| dc.date.accepted | 2025-02-11 | - |
| dc.contributor.author-college | 理學院 | - |
| dc.contributor.author-dept | 化學系 | - |
| dc.date.embargo-lift | 2030-02-10 | - |
| 顯示於系所單位： | 化學系 | |
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