臺灣博碩士論文加值系統

在生化結構研究上發現由產毒性大腸桿菌(ETEC)所分泌的忌熱型腸毒素(LT)與霍亂弧菌所分泌的霍亂毒素(CT)結構有非常大的相似性，對於毒素的作用機轉之闡明已經可以達到分子甚至原子如此細微的等級。毒素五元的次單元B會辨認人類小腸上皮細胞GM1接受器，並引發細胞的內吞作用(endocytosis)而將毒素帶入細胞內，毒素的A1蛋白片段可使α刺激型Ｇ蛋白(Gsα)與鳥嘌呤核苷三磷酸(GTP)持續維持在結合狀態，此複合物會活化腺核甘環狀酶(adenyl cyclase) 造成環狀腺核苷單磷酸(cAMP)不斷的被製造，使cAMP濃度上升，造成細胞內離子及水份大量流出至腸腔，引起水瀉症狀產生。
對於抑制毒素次單元B接受體的化學結構設計上，延續先前的研究成果，以2-[(4-methoxybenzyl)oxy]benzoic acid (31)為先導化合物，合成了一系列2 (or 3 or 4)-substituted benzyloxy benzoic acids、4 (or 5 or 6)-methoxy-2-substituted benzyloxy benzoic acids、4-substituted benzyloxy isophthalic acids、1 or 3-substituted benzyloxy -2-naphthoic acids、 2-substituted benzyloxy-1-naphthoic acids以及針對benzoic acid羧基延長的化合物。
在生物活性篩選上我們藉由GM1-酵素結合免疫吸附法(GM1-ELISA)測定活性，並以老鼠做毒素下痢分析的動物實驗，其結果顯示4 or 6-methoxy-2- substituted benzyloxy benzoic acids及3-substituted benzyloxy benzoic acids 具有明
顯的抗下痢活性，值得更進一步研究。

Structural biology studies on heat-labile enterotoxin (LT) from enterotoxigenic Escherichia coli. and the closely related cholera toxin (CT), shed light on the action mechanism of toxin at molecular and atomic levels. The B pentamer protein of the toxins recognized the receptor ganglioside GM1 presented on the gastrointestinal tract of human host and triggers endocytosis . The enzymatic A1 fragment of the toxin enters the cytosol it modifies the alpha subunit of stimulatory G protein (Gsα) and locks the G protein in its GTP-bound form, which continually stimulates adenylate cyclase to produce cAMP. The resulting elevate levels of cAMP cause dramatic efflux of ions and water from the host, leading to watery diarrhea.
Structure-based design has led to various classes of compounds targeting toxin B subunit bonding site in our Laboratory. Following the previous results, 2-[(4-methoxybenzyl)oxy]benzoic acid (31) was used as a lead compound, a series of 2 (or 3 or 4)-substituted benzyloxy benzoic acids, 4 (or 5 or 6)-methoxy-2-substituted benzyloxy benzoic acids, 4-substituted benzyloxy isophthalic acids, 1 or 3-substituted benzyloxy-2-naphthoic acids, 2-Substituted benzyloxy-1-naphthoic acids and carboxyl group extend of benzoic acids were synthesized.
The biological activities of these syntheted compound were examined by GM1 Enzyme-linked Immunosorbent Assay (GM1-ELISA) and Patent Mouse Gut Assay, the result showed 4 or 6-methoxy-2-substituted benzyloxy benzoic acids and 3-substituted benzyloxy benzoic acids have significant anti-diarrhea activities and worthy for further investigation.

中文摘要
英文摘要
第一章 緒論
第一節 背景資料 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 1
第二節 小腸上皮細胞氯離子分泌之調控機轉 ‥‥‥‥‥‥‥‥ 3
第三節 目前治療嚴重細菌性腹瀉的方法 ‥‥‥‥‥‥‥‥‥‥ 5
第四節 忌熱型腸毒素(heat-labile enterotoxin; LT)結構解析 ‥‥‥ 7
第五節 忌熱型腸毒素致病機轉 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 10
第六節 相關研究現今概況 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 17
第七節 如何看出立體圖形？ ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 21
第八節 研究動機與目的 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 23
第二章 標的化合物之設計與化學合成
第一節 化合物之設計 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 26
第二節 產物構造決定
一、2-Substituted Benzoic acids 類化合物構造決定 ‥‥‥‥‥‥ 35
二、3-Substituted Benzoic acids 類化合物構造決定 ‥‥‥‥‥‥ 41
三、4-Substituted Benzoic acids 類化合物構造決定 ‥‥‥‥‥‥ 46
四、4-Methoxy-2-Substituted Benzoic acids 類化合物構造決定 ‥ 49
五、5-Methoxy-2-Substituted Benzoic acids 類化合物構造決定 ‥ 54
六、6-Methoxy-2-Substituted Benzoic acids 類化合物構造決定 ‥ 59
七、4-Substituted isophthalic acids 類化合物構造決定 ‥‥‥‥‥ 64
八、1-substituted-2-naphthoic acids 類化合物構造決定 ‥‥‥‥ 67
九、3- substituted-2-naphthoic acids 類化合物構造決定 ‥‥‥‥ 73
十、2-Substituted-1-naphthoic acids 類化合物構造決定 ‥‥‥‥ 79
十一、(3E)-4-{2-[(substituted)oxy]-phenyl}but-3-en-2-ones 類化合物構造決定 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 85
十二、3-{2-[(substituted)oxy]-phenyl}propanoic acids 類化合物構造決定 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 91
第三節 試藥、溶媒及材料 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 97
第四節 重要儀器 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 101
第五節 化合物之製備
一、實驗方法改良及注意事項 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 103
二、Methyl substituted benzoate 類化合物之合成 ‥‥‥‥‥‥ 113
三、Substituted benzoic acid 類化合物之合成 ‥‥‥‥‥‥‥ 134
四、Methyl methoxy substituted benzoate 類化合物之合成 ‥‥ 155
五、Dimethyl 4-substituted isophthalate 類化合物之合成 ‥‥‥ 176
六、Methyl substituted naphthoat 類化合物之合成 ‥‥‥‥‥‥ 183
七、Methoxy substituted benzoic acid 類化合物之合成 ‥‥‥‥ 204
八、4-Substituted isophthalic acid 類化合物之合成 ‥‥‥‥‥‥ 222
九、Naphthoic acid substituted 類化合物之合成 ‥‥‥‥‥‥‥ 228
十、(3E)-4-{2-[(substituted)oxy]-phenyl}but-3-en-2-ones類化合物之合成 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 253
十一、(2E)-3-{2-[(substituted)oxy]-phenyl}acrylic acids類化合物之合成 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 259
十二、3-{2-[(substituted)oxy]-phenyl}propanoic acid類化合物之合成 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 271
十三、Nicotinic acid類化合物之合成 ‥‥‥‥‥‥‥‥‥‥‥ 284
第三章 藥理活性試驗
第一節 抗下痢活性試驗方法 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 287
第二節 藥理活性試驗結果
一、GM1 – LTB ELISA ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 289
二、劑量與活性間關係（Dose Responsive）‥‥‥‥‥‥‥‥‥ 292
三、Patent Mouse Gut Assay ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 295
第四章 結論
結構與活性關聯性之探討 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 296
參考文獻 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 300
圖譜解析 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 306
索引表 ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 658

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