臺灣博碩士論文加值系統

環狀胜肽因其良好的親和性和選擇性，可應用於一些疾病的治療，例如皮膚癌，然而許多環狀胜肽在自然環境中存量不足，難以取得，因此目前研究致力於發展以新的合成方式獲得環狀胜肽。Growth-factor-bound protein 2（Grb2）– Src homology 2（SH2）在Ras信號傳導路徑佔有極為重要的地位，這個訊號通路異常恐造成腫瘤的發生，近期環狀胜肽被報導出對於可作為Grb2-SH2的抑制藥劑，為一種從噬菌體中萃取出的具有雙硫鍵的環十一肽disulfide-bridged cyclic peptide（G1），然而因其結構中的雙硫鍵不穩定，為了解決這個問題，有了許多後續的研究，許多研究將雙硫鍵置換為較穩定的結構。此外並有研究顯示其環狀胜肽的雙環相似物有更佳的活性，本研究設計了雙環胜肽的類似物並進行其胜肽分子的合成研究，其中先利用固相合成獲得目標的胜肽序列，並以高效液相色譜法進行純化，接著在水相進行烯烴複分解反應獲得環狀胜肽。

Due to the good affinity and selectivity of cyclic peptides towards their targets, these have been reported to be therapeutic agents that can be used in treatment for many types of diseases, such as skin cancer. However, cyclic peptides are difficult to obtain from the nature. Currently, more and more synthetic methodology are discovered to find new synthetic methods to synthesize cyclic peptides. Growth-factor-bound protein 2 (Grb2) – Src homology 2 (SH2) plays an important role in the Ras signaling pathway. Any abnormal signals in the pathway might indicate tumorigenesis. Recently, cyclic peptides have been reported to be a new type of therapeutic agent for inhibition of Grb2-SH2 domain. G1, a disulfide-linked 11-residue cyclic peptide, is an inhibitor of Grb2-SH2 at micromolar range. However, it is very unstable under physiological conditions because of instability of the disulfide bond. In order to solve this problem, there are many research to replace the disulfide bond for stable structure. In addition, there are also some studies that demonstrated bicyclic peptides analogues have better biological activity. Therefore, in this thesis, an analogue of bicyclic peptides has been designed and synthesized. The targeted of peptide sequence is obtained by solid phase peptide synthesis, and purified by high performance liquid chromatography followed by peptide cyclization using ring-closing metathesis under aqueous conditions.

論文審定書 i
論文公開授權書 ii
致謝 iii
中文摘要 iv
Abstract v
目錄 vi
圖目錄 viii
表目錄 ix
縮寫表 x
第一章 序論 1
1.1 環狀胜肽應用 1
1.1-1 Ras信號傳導路徑 2
1.1-2 Grb2-SH2抑制劑 4
1.2 胜肽合成 8
1.2-1 樹脂固相合成載體 9
1.2-2 固相多肽合成（SPPS） 11
1.2-3 微波輔助多肽合成 12
1.2-4 紫外-可見分光光度法定量分析 13
1.2-5 串聯式質譜儀（MS/MS） 14
1.3 環狀胜肽的合成 16
1.3-1 閉環複分解反應 16
1.3-2 水相烯烴複分解反應 17
第二章 研究動機 19
第三章 結果與討論 22
3.1 胜肽的合成 22
3.1-1 FmocSacOH（8）的合成 22
3.1-2 FmocOasOH（14）的合成 23
3.1-3 多肽固相合成（solid phase peptide synthesis） 24
3.2 環狀胜肽的合成 41
3.2-1 水相烯烴複分解反應 41
第四章 結論 44
第五章 引用文獻 45
第六章 儀器設備與藥品材料與實驗步驟 48
6.1 儀器設備與藥品材料 48
6.2 合成步驟 52
6.2-1 BocCysOMe（5） 52
6.2-2 BocSacOMe（6） 54
6.2-3 FmocSacOH（8） 55
6.2-4 BocSerOH（9） 57
6.2-5 Allyl tert-butyl carbonate（10） 58
6.2-6 Allyl O-allyl-N-（tert-butoxycarbonyl）-L-serinate（11） 59
6.2-7 BocOasOH（12） 61
6.2-8 FmocOasOH（14） 62
6.2-9 固相胜肽合成 64
6.2-10 高效液相層析儀純化胜肽16 75
6.2-11 水相烯烴複分解製備環狀胜肽 77
第七章 光譜與圖譜資料 79

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