環狀胜肽逐漸成為一種新類型的治療藥劑，在醫學上已有利用於抗菌、皮膚癌與移植器官的抑制劑。本篇論文是研究對於Grb2-SH2蛋白有抑制效果的環狀胜肽，因為Grb2-SH2對於Ras蛋白的訊號傳遞有相當大的影響。

Grb2-SH2的天然受質為磷酸化酪氨酸，而2003年Roller教授究團隊發現一種含有10個胺基酸的環狀胜肽，可以抑制Grb2-SH2的活性，其序列的穀胺酸2、酪氨酸4、天門冬醯胺6能夠以仿磷酸化酪氨酸的形式與Grb2-SH2鍵結。

相較於Roller團隊的環狀胜肽，本論文在胜肽分子的設計上保留了3個關鍵胺基酸的序列，使目標環狀胜肽由七個胺基酸組成。在合成環狀胜肽所採用的方法，是相較液相合成快速方便且容易純化的固相合成，並利用Kaiser 試劑檢測每一段胺基酸都成功的鍵結上17，所得的鏈狀胜肽，經由閉環複分解的方式，進行合環的反應。未來我們實驗室將經由活性檢測，進而探討縮短後的胜肽分子是否也能有抑制效果。

Cyclic peptides have gradually became a new type of therapeutic agent in medicine. They have been useful in treatment of bacCyclic peptides have gradually became a new type of therapeutic agent in medicine. They have been useful in treatment of bacterial infection, skin cancer and as organ transplant inhibitors. This thesis focus on design, synthesis and biological activity of a cyclic peptide for the inhibition of Grb2-SH2. Grb2-SH2 plays an important role in Ras signaling pathway.
The natural substrate for Grb2-SH2 is phosphotyrosine. In 2003, Roller and his team had reported a 10-residue cyclic peptide that could inhibit Grb2-SH2. It was found that Glu2, Tyr4 and Asn6 act as transition state analogue for phosphotyrosine during binding with Grb2-SH2.
In this thesis, an analogue of this cyclic peptide was designed and synthesized. Compared with the peptide found by Roller’s team, the sequence of the target peptide in this thesis retained the 3 key residues while the rest was truncated leading to a 7-residue cyclic peptide. The linear peptide was synthesized by solid-phase peptide synthesis and purified by HPLC. Finally, the desired cyclic peptide could then be obtained with ring closing metathesis in water. In the future, our laboratory will examine biological activity of the truncated peptide.

論文審定書 i
論文公開授權書 ii
誌謝 iii
中文摘要 iv
英文摘要 v
目錄 vi
圖目錄 viii
表目錄 x
縮寫表 xi
第一章緒論 1
第一節 環狀胜肽的應用 1
1-1.Tyrocidine和短桿菌肽S (gramicidin S) 的抗菌活性 1
1-2.環孢素A的免疫抑制活性 2
1-3.RGD胜肽的抗血管生成活性 3
第二節 胜肽合成 3
2-1.活化基團 4
2-2.液相合成 5
2-3.固相合成載體 5
2-4.固相合成 6
2-5.微波加熱方式合成 7
第三節 環狀胜肽的合成 7
3-1.合成方式 7
3-2.利用閉環複分解 (Ring Closing Metathesis ) 合環 8
3-3.MSMS原理 9
第四節 目標胜肽的介紹 11
4-1.生物活性原理 11
4-2.胜肽設計的概念 11
4-3.逆合成分析 12
第五節 研究動機 12
第二章 結果與討論 14
第一節 起始物的合成 14
2-1.以Fmoc為保護基的合成N-Fmoc-S-Allyl-Cysteine tert-butyl ester (4) 14
2-2.以Boc為保護基的合成N-Boc-S-Allyl-cysteine methyl ester (BocSacOMe) (8) 15
2-3.N-Fmoc-S-Allyl-Cysteine (FmocSacOH) (11) 的合成 16
3.固相合成SPPS 17
4.MS-MS圖譜分析 19
5.閉環複分解RCM 21
第三章 結論 23
第四章 引用文獻 24
第五章 儀器設備與藥品材料與實驗步驟 26
第一節 儀器設備與藥品材料 26
第二節 合成步驟 29
第三節 Solid Phase Peptide Synthesis 38
第四節 利用閉環複分解合環合成化合物 13 40
第六章 光譜與圖譜資料 41

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