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標題: | 鑑定與探討異柴胡內酯活性標靶及其引發3-磷酸甘油醛去氫酶級聯反應於治療去勢療法無效之前列腺癌 Elucidation of Isochaihulactone-targeted Protein and its Applications of Using GAPDH Cascade for Castration-resistant Prostate Cancer Treatment |
作者: | Jeffy Chern 陳剛健 |
指導教授: | 吳世雄(Shih-Hsiung Wu) |
關鍵字: | 異柴胡內酯,活性探針,去勢療法無效之前列腺癌,3-磷酸甘油醛去氫?級聯反應,雄激素受體,E3-泛蛋白連接?, isochaihulactone,activity-based probe,castration-resistant prostate cancer,GAPDH cascade,androgen receptor,Siah1, |
出版年 : | 2018 |
學位: | 博士 |
摘要: | 傳統天然藥物在做為尋找具生物活性的前導化合物上,扮演著舉足輕重的角色,但它們要發展成為現代藥物的過程中,特別是鑑定其活性標靶卻充滿困難。本論文欲探討Z-(+)-異柴胡內酯(一種萃取自南柴胡Bupleurum scorzonerifolium的木質素)的活性標靶蛋白及其分子機轉,用於後續藥物開發的潛力。首先,我們發現異柴胡內酯可以顯著地抑制一些具抗藥性之前列腺癌細胞株及其衍生之異體移植小鼠腫瘤的生長。藉由與硫醇鹽(thiolate)反應,並以核磁共振光譜分析得知,異柴胡內酯對於生物內生性的親核基團(nucleophile)不具有偏離目標性(off-targeting),表示此化合物為標靶共價結合抑制劑(targeted covalent inhibitor)。通過鑑定藥效基團發現,異柴胡內酯上的α,β-不飽和基團是活性中心。因此,利用此一特性,我們利用活性探針為基礎的蛋白質體分析,鑑定出3-磷酸甘油醛去氫酶為其活性標靶。之後,利用液相層析串聯式質譜及電腦輔助分子嵌合模擬闡明,3-磷酸甘油醛去氫酶的非催化半胱胺酸-247與柴胡內酯的共價產物扮演著細胞凋亡與配體結合區(LBD)非依賴性雄激素受體降解的「開/關」。這些研究成果將有助於改善現今在去勢療法無效之前列腺癌所面臨無藥可醫的窘境。 Traditional medicines provide a fertile ground to explore potent 'lead' compounds with desired effects; yet, their transformation into modern drugs is often fraught with challenges in deciphering the target that is mechanistically valid for its biological activity. This dissertation seeks to determine the mechanistic target of Z-(+)-isochaihulactone 1 (active lignin from a traditional Chinese medicine - Nan Chai Hu, Bupleurum scorzonerifolium) and whether compound 1 can serve as a promising lead compound for future drug development. Firstly, we revealed that compound 1 markedly inhibits the growth of some drug-resistant prostate cancer cell lines and mice xenografts. By NMR spectroscopy, compound 1 was shown to resist to an off-targeting thiolate, thus giving compound 1 a targeted covalent inhibitor-like (TCI) property. By identifying the pharmacophore of compound 1 (α,β-unsaturated moiety), a compound 1-derived probe was designed and synthesized for TCI-oriented activity-based proteome profiling (TCI-ABPP). With MS/MS and computer-guided molecular biology approach, we elucidated that the noncatalytic C247 of GAPDH was indispensable in controlling the “ON/OFF” switch of apoptosis and ligand binding domain (LBD)-independent androgen receptor (AR) degradation. Taken together, these findings shed new light on the obstacle existed in castration-resistant prostate cancer (CRPC) treatment. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/7595 |
DOI: | 10.6342/NTU201800828 |
全文授權: | 同意授權(全球公開) |
電子全文公開日期: | 2028-05-18 |
顯示於系所單位: | 化學系 |
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ntu-107-1.pdf 此日期後於網路公開 2028-05-18 | 16.21 MB | Adobe PDF |
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