臺灣博碩士論文加值系統

中文摘要
利用鈀金屬催化交叉耦合反應合成腔腸螢光素的類似物可用在研究鳶烏賊體內螢光蛋白的發光機制上。薗頭交叉耦合反應可在2-氨基吡嗪的五號位置引入乙炔基。在縮合反應中，使用4-甲基苯磺酸吡啶鹽可讓對酸敏感的乙炔基不受破壞而得到類似物153和160。相較於天然腔腸螢光素149和15，此二類似物的發光波譜可觀察到有助於螢光蛋白發光機制研究的紅位移。而乙炔基的引入使取代基長度增加亦可探討螢光蛋白對類似物的接受度。
腔腸螢光素氧化後的醯胺化合物168a是生物螢光發中機制中的發光器。藉由合成168b-d並量測其發光波譜，可發現類似物的放光相較於天然腔腸螢光素大約有六十至八十奈米的紅位移。




Abstract
Many coelenterazine analogs have been synthesized via palladium-mediated cross coupling reactions for bioluminescent studies toward photoprotein Symplectin. Instead of hydrochloric acid, pyridinium p-toluenesulfonate (PPTS) provides a method to obtain coelenterazine analogs containing acid-sensitive moieties. After this method was developed, the analogs having acetylene moieties at 6-position of coelenterazine was synthesized via Sonogashira cross coupling. Analogs 153 and 160 both emit light at longer wavelength compared to natural coelenterazine 149 and 15 which are suitable for protein studies. Besides, compound 153 and 160, which having longer distance at 6-position, provide a way to exam the aceptability of Symplectin.
As light emitter in bioluminescent mechanism, coelenteramide analogs (168a-d) were synthesized. It is confirmed that our synthetic analogs have potential to emit luminescence 60~80 nm higher wavelength compared with the natural coelenterazine.

Index
中文摘要 i
Abstract ii
Acknowledgement iii
Index iv
Figure index vii
Table index ix
Equation index ix
Scheme index x
Abbreviations xii

Chapter 1 Introduction 1
1.1 General introduction of bioluminescence 1
1.1.1 Firefly 2
1.1.2 Cypridina luciferin 3
1.1.3 Photo protein, Aequorin 4
1.1.4 Luminous squid, Symplectoteuthis oualaniensis 5
1.2 Literature review on the synthesis of Coelenterazine and Dehydrocoelenterazine 7
1.3 Literature review on the biological studies toward Symplectoteuthis oualaniensis 18
1.4 Research of our group 26
1.5 Proposal of this study 29

Chapter 2 Synthesis of coelenterazine analogs 32
2.1 Bromobenzylaminopyrazine 32
2.2 Terminal acetylene preparation 33
2.3 3,5-unidentical disubstitued 2-aminopyrazine 36
2.3.1 Synthesis of natural form coelenteramine 36
2.3.2 Synthesis of acetylene-inserted coelenteramine analogs 38
2.4 Condensation reaction of ketoacetal and coelenteramine 39
2.5 Synthesis of coelenteramide 43


Chapter 3 Spectroscopic studies of coelenterazine analogs 48
3.1 Coelenteramine 49
3.2 Coelenterazine 56
3.2.1 Extinction coefficient 56
3.2.2 Effect of pH to the absorption spectra 61
3.2.3 Fluorescence - emission spectra of coelenterazines 65
3.3 Coelenteramide 69
3.4 Chemiluminescence 74

Chapter 4 Conclusion 78
4.1 Conclusion 78
4.2 Future prospect 81

Chapter 5 Experimental Section 82
5.1 General Methods 82
5.2 Experimental procedures and analytical data 84
5.2.1 Dibromination (122) 27 84
5.2.2 Negishi coupling (119) 13 85
5.2.3 Sonogashira coupling (131) 86
5.2.4 Terminal acetylene preparation (132) 87
5.2.5 O-acetyl protection (133) 88
5.2.6 Sonogashira coupling (134) 89
5.2.7 Desilylation (135) 90
5.2.8 Suzuki coupling (139) 91
5.2.9 Demethylation (140) 92
5.2.10 Sonogashira coupling (141) 93
5.2.11 Sonogashira coupling (142) 94
5.2.12 Silyl ether protection (143) 95
5.2.13 Condensation for natural form coelenterazine (149) 96
5.2.14 Condensation for coelenterazine analog (153) 97
5.2.15 Demethylation of coelenterazine (154) 98
5.2.16 Condensation for coelenterazine analog (160) 99
5.2.17 Condensation for natural form coelenterazine (154) 100
5.2.18 Model study for amide bond formation (166) 101
5.2.19 Natural form coelenteramide (168a) 102
5.2.20 Demethylation of coelenteramide (168e) 103
5.2.21 Coelenteramide analog (168d) 104
5.2.22 Coelenteramide analog (168b) 105
5.2.23 Coelenteramide analog (168c) 106
5.2.24 Oxidation to afford dehydrocoelenterazine (187) 107
5.2.25 Preparation of ketoacetal (159) 16a 108

Chapter 6 References 109
Supporting information 112

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31. http://web.uvic.ca