臺灣博碩士論文加值系統

許多具功能的金屬蛋白都有金屬離子做為輔酶。因此，移除金屬離子即有潛力成為抑制金屬蛋白功能的方法。金屬螯合劑是一種化學製劑，不僅可以螯合金屬離子移出體外，也能螯合金屬蛋白內的金屬離子，改變金屬蛋白的功能。在過去十年間，很多新穎的螯合劑被發展出來，他們被用來抗病毒，抗腫瘤，抗纖維化，與做為光保護劑。二乙胺四乙酸是已知強效金屬離子螯合劑，可清除大多數金屬離子。因此，我們仿照類似二乙胺四乙酸羧基基團的金屬離子螯合劑，設計並合成雙丙二酸及雙-(二乙酸)鈉鹽衍生物。合成雙丙二酸鈉鹽衍生物是以4-羥基苯甲醛當起始物，先與丙二酸二乙酯反應生成雙丙二酸二乙酯衍生物，再進行水解得到雙丙二酸鈉鹽衍生物。合成雙-(二乙酸) 鈉鹽衍生物是由4-亞硝基苯酚鈉鹽當起始物，與雙鹵烷類反應成雙硝基衍生物，之後再將雙硝基還原成雙胺基衍生物。最後，再與溴乙醯乙酯反應形成胺基雙取代。產物再進行水解得雙-(二乙酸) 鈉鹽終產物。將合成的13個新穎產物進行亞鐵離子螯合試驗、抑菌、整合脢活性抑制試驗。實驗結果顯示4e, 5e, 6c, 8d, 9d, 12d, and 13d在亞鐵離子螯合試驗中具有相當好的螯合力(CE50 = 0.72 – 8.78 mM)。抗菌實驗中，全部化合物並無顯著的抑制。在愛滋病毒整合脢活性抑制試驗中，化合物1e, 5e, 7d, 8d, 9d, 11d, 12d, 13d對整合脢活性具有極性的抑制效果(IC50 = 0.8 ~19.1μM)。

Metal ions are essential cofactors for many functional metalloproteins. Thus, removel of metal ions may be a useful way to alter the function of metalloproteins. Metal ions chelators are a class of chemical agents that are able not only to bind and remove metal ions from the body, even from metalloproteins. In the past decade, many new viable utilities for metal ions chelators have been reported. This includes the use of iron chelators as antiviral, antiproliferative, antifibrotic, and photoprotective. EDTA has been shown to be a powerful chelators, which chelates a large range of metal ions. For these facts, we designed and synthesized bis-(di-acetic acid) and bis-malonic acid sodium salt derivatives with similar carboxylic acid residue to EDTA as new potential metal ions chelators. Synthesis to bis-malonic acid sodium salt derivatives started from 4-hydroxybenzaldehyde, which was first reacted with diethylmalonate to give diethylmalonate derivates. Hydrolysis of the ester provided bis-malonic acid sodium salts. Synthesis of bis-(di-acetic acid) sodium salt derivatives, started from sodium 4-nitrophenolate, which was reacted with dibromoalkane afford to the dinitro derivatives. The dinitro was reduced to the diamino derivatives, which was further converted to amino-disubstituents. Finally, the amino disubstituents were hydrolyzed to bis-(di-acetic acid) sodium salt. The prepared 13 novel products were assayed for their effects on ferrous-chelation test, anti-bacteria, collagen degradation assay, and HIV-1 integrase inhibition test. Preliminary results showed 4e, 5e, 6c, 8d, 9d, 12d, and 13d possessed good ferrous chelation (CE50 = 0.72 – 8.78 mM). In antibacterial test, no inhibitory activity was showen. In HIV-1 integrase inhibitory test, 1e, 5e, 7d, 8d, 9d, 11d, 12d, 13d possessed good HIV-1 integrase inhibitory activity(IC50 = 0.8 ~19.1μM).

目錄

頁
正文目錄……………………………………………………………………...… I
附表目錄……………………………...………………………………………...Π
圖目錄…………………………………….......………………………..………III
附圖目錄………………………………………………………………….Ⅳ、Ⅴ
中文摘要……………………………………………………………………….Ⅵ
英文摘要………………………………………………………………..….......Ⅶ

正文目錄

第一章 緒論…………………………………...……………………………..…1
第一節 研究背景………………………………………...………………….1
壹、人體必要元素……………………………………….………………..2
貳、金屬離子蓄積與金屬離子螯合劑…………..………..……….……..6
參、鐵離子過量蓄積及非鐵離子過量蓄積造成的疾病…………..……7
肆、自由基簡介…………………………………….………………...…13
伍、螯合理論…………………………………….………………...……16
陸、螯合劑簡介………………………………….………………...…...20
第二節 研究目的…………………………………………………...……...26
壹、 研究動機……………………………….………………...………26
貳、 合成反應概要……………………………………………………30
第二章 材料與方法………………………………………...…………………36 第一節 試劑、溶媒及原料……………………………………...………...36
第二節、重要儀器……………………………………………….…………38
第三節、合成實驗……………………………………….…………………40
第四節、生物活性部分……………………………….……………………83
第三章 結果……………………………………………………………...……87
第四章 討論…………………………………………………………………...93
第五章 結論………………………………………………..….…………..…..98
第六章 參考文獻………………………………………………………...……99








附表目錄

表1、人體必要金屬元素……………………………...………………………..5
表2、金屬離子與螯合物之酸鹼理論分類表………….…………………..…18
表3、常見金屬離子與螯合配位基團之幾何形式對應表…………………...18
表4、常見人工合成的螯合劑………………………………………………...23
表5、化合物螯合亞鐵離子能力.....................................88
表6、化合物螯合亞鐵離子的能力評估..................................90
表7、合成終產物對五種細菌的抑菌活性評估………………………….…...91
表8、合成終產物對愛滋病毒整合脢的抑制活性評估………………………92




圖目錄

圖1、HIV 整合脢三大功能區位………………….……………..…………….10
圖2、HIV 整合脢之胺基酸序列…………………………………………..…..10
圖3、具芳香羥基HIV整合脢抑制劑之抑制機轉.…………………………..11
圖4、含catechol結構之HIV-1整合酶抑制劑………………...……………12
圖5、環狀螯合圖示………………..………….………………………………16
圖6、螯合劑對金屬離子之氧化還原電位............................19
圖7、catechol-type……………………………………………………….…21
圖8、hydroxamic acid-type……………………………….………………...21
圖9、細菌螯合鐵的過程….…………………………………………………..22
圖10、局部光照保護螯合劑……….…………………………………………..24
圖11、創傷處理及抑制HIV病毒螯合劑….……….……………………...…25
圖12、螯合劑….………………………………………………...………..……27
圖13、具雙酮結構的化合物………………………......……………………...28
圖14、對稱化合物與HIV-1 Integrase電腦分子模擬………....….……….29
圖15、對稱螯合基團化合物………………………...……………..…………29
圖16、合成化合物1e、2e、3e、4e、5e、6c及對照組Vit. C及EDTA分別以0、0.2、1、2、4、5、10、20、30 mM進行作圖...................................89
圖17、合成化合物7d、8d、9d、10d、11d、12d、13d及對照組Vit. C及EDTA分別以0、0.2、1、2、4、5、10、20、30 mM進行作圖............................90




附圖目錄

附圖1、化合物1b及1c 1H NMR圖譜……………………………………104
附圖2、化合物1d及1e 1H NMR圖譜…………………………………….105
附圖3、化合物1e 13C NMR及2b 1H NMR圖譜…….…………………..106
附圖4、化合物2c及2d 1H NMR圖譜……………………………………107
附圖5、化合物2e 1H NMR及 13C NMR圖譜…………………..….…….108
附圖6、化合物3b及3c 1H NMR圖譜……………………….….….…….109
附圖7、化合物3d及3e 1H NMR圖譜……………………………..……..110
附圖8、化合物3e 13C NMR及4b 1H NMR圖譜………...………..……..111
附圖9、化合物4c及4d 1H NMR圖譜………...……………….……..…..112
附圖10、化合物4e 1H NMR及 13C NMR圖譜……………………….….113
附圖11、化合物5b及5c 1H NMR圖譜…………………………………..114
附圖12、化合物5d及5e 1H NMR圖譜………………………….….…….115
附圖13、化合物5e 13C NMR 及6b 1H NMR圖譜…………...….………..116
附圖14、化合物6c 1H NMR及 13C NMR圖譜…………………….….….117
附圖15、化合物7b及7c 1H NMR圖譜…………………………………..118
附圖16、化合物7d 1H NMR及 13C NMR圖譜…………………………..119
附圖17、化合物8b及8c 1H NMR圖譜…………………………….…….120
附圖18、化合物8d 1H NMR及 13C NMR圖譜………………………….121
附圖19、化合物9b及9c 1H NMR圖譜………………………………..…122
附圖20、化合物9d 1H NMR及 13C NMR圖譜……………………….....123
附圖21、化合物10b及10c 1H NMR圖譜………………………………..124
附圖22、化合物10d 1H NMR及 13C NMR圖譜…………………………125
附圖23、化合物11b及11c 1H NMR圖譜………………………………..126
附圖24、化合物11d 1H NMR及 13C NMR圖譜…………………………127
附圖25、化合物12b及12c 1H NMR圖譜………………………….....….128
附圖26、化合物12d 1H NMR及 13C NMR圖譜…………………..……..129
附圖27、化合物13b及13c 1H NMR圖譜……………………...…………130
附圖28、化合物13d 1H NMR及 13C NMR圖譜………………...……….131

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