| Strecker合成法合成降冰片烯基甘氨酸及与四嗪的生物正交反应 |
| Alternative Title | Strecker synthesis of 2-amino-2-(bicyclo[2.2.1]hept-5-en-2-yl)acetic acid and bioorthogonal reaction with tetrazine
|
| 张紊玮 |
| Thesis Advisor | 董守良
|
| 2010-05-20
|
| Degree Grantor | 兰州大学
|
| Place of Conferral | 兰州
|
| Degree Name | 硕士
|
| Keyword | 降冰片烯
四嗪
Strecker合成法
生物正交反应
逆电子需求狄尔斯—阿尔德反应
|
| Abstract | 目的：降冰片烯与四嗪能够发生逆电子需求狄尔斯—阿尔德反应，放出氮气最终生成二氢哒嗪，符合典型的生物正交反应。与以往的生物正交反应相比，反应具有更高速率、无需任何催化剂、无毒性、底物合成方法简单等特点，但是，还未曾在体内/外标记多肽或蛋白中崭露头角。若将降冰片烯基整合到氨基酸中，进而整合到多肽或蛋白中，再与四嗪进行专一性结合，那么可以实现在生物体内/外对多肽或蛋白的标记。
方法：运用斯特雷克氨基酸合成法 (Strecker synthesis)合成5-降冰片烯基-2-甘氨酸；用肾酰基转移酶 I 分离L-5-降冰片烯基-2-甘氨酸；合成3, 6-二苯甲酸-1, 2, 4, 5-四嗪、3-苄胺-1, 2, 4, 5-四嗪，研究与5-降冰片烯基-2-甘氨酸的环化加成反应；将5-降冰片烯基-2-甘氨酸Fmoc保护，采用Fmoc固相合成法将5-降冰片烯基-2-甘氨酸整合到抹香鲸肌红蛋白一段五肽中，在PBS缓冲液中用四嗪进行标记；
结果与结论：通过Strecker氨基酸合成法，方便合成5-降冰片烯基-2-甘氨酸，总产率41%；用肾酰基转移酶 I分离L-5-降冰片烯基-2-甘氨酸，产率21%；合成3, 6-二苯甲酸-1, 2, 4, 5-四嗪、3-苄胺-1, 2, 4, 5-四嗪，产率分别为67%和23%；用Fmoc固相合成法将5-降冰片烯基-2-甘氨酸整合到抹香鲸肌红蛋白一段五肽中，在PBS缓冲液中用3-苄胺-1, 2, 4, 5四嗪进行了专一性标记。 |
| Other Abstract | Objective: Norbornene reacts with tetrazine forming dihydropyridazine products and dinitrogen based on inverse electron demand Diels-Alder reaction. The reaction meets bioorthogonal reaction criteria. Comparing with former bioorthogonal reactions, tetrazine-based cycloadditions has more features such as higher rates, without need of any catalyst, non-toxicity and readily available starting materials, but it still not emerging in labeling peptides or proteins in vivo/in vitro. If norbornene was incorporated into amino acid，and then integrated into peptides or proteins which then were labeled with tetrazine. So we could localize target peptides or protein in vivo/in vitro specifically.
Methods: 5-norbornene-2-glycine was synthesized via Strecker synthesis; Kidney acylase I was used to isolate L-5-norbornene-2-glycine from DL-5-norbornene-2-glycine; 3,6-dibenzoic acid-1,2,4,5-tetrazine and 3-benzylamino-1,2,4,5-tetrazine were synthesized separately, then study the cycloaddition reactions between 5-norbornene-2-glycine and tetrazine; integrate 5-norbornene-2-glycine (Fmoc protected) into pentapeptide of sperm whale myoglobin and label pentapeptide using tetrazine in PBS buffer.
Results and Conclusion: We synthesized 5-norbornene-2-glycine through Strecker reaction with total yield of 41%, separated L-5-norbornene-2-glycine using Kidney acylase with yield of 21%; Then we synthesized 3,6-dibenzoic acid-1,2,4,5-tetrazine and 3-benzylamino-1,2,4,5-tetrazine with yield of 67% and 23%, respectively; We used Fmoc solid phase peptide synthesis intergrated Fmoc-5-norbornene-2-glycine into pentapeptide of sperm whale myoglobin and labeled pentapeptide using 3-benzylamino-1,2,4,5-tetrazine in PBS buffer. |
| URL | 查看原文
|
| Language | 中文
|
| Document Type | 学位论文
|
| Identifier | https://ir.lzu.edu.cn/handle/262010/221848
|
| Collection | 生命科学学院
|
Recommended Citation
GB/T 7714 |
张紊玮. Strecker合成法合成降冰片烯基甘氨酸及与四嗪的生物正交反应[D]. 兰州. 兰州大学,2010.
|
Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.
