臺灣博碩士論文加值系統

在本研究中我們以ㄧ壺合成法 (one-pot synthesis) 合成了八個異黃酮化合物，並且測試這些異黃酮化合物對蕈類酪胺酸酶的抑制效果及 B16F10細胞存活率試驗。
這些異黃酮類化合物為以A環合併B環具有不同取代基作為基礎的7,8-二羥基異黃酮類化合物和5,7-二羥基異黃酮類化合物。7,8-二羥基異黃酮類化合物有：7,8-Dihydroxyisoflavone (1)、7,8- Dihydroxy-
4''-methoxyisoflavone (2)、7,8-Dihydroxy- 3''- methoxyisoflavone (3) 及3'',7,8-Trihydroxyisoflavone (4)。5,7-二羥基異黃酮類化合物有：5,7-Dihydroxyisoflavone (5)、5,7-Dihydroxy-4''-methoxyisoflavone (6)
、5,7-Dihydroxy-3''-methoxyisoflavone (7)及3'',5,7-Trihydroxyisoflavone (8)。所有的化合物結構皆經由光譜數據及文獻資料鑑定其結構。
對蕈類酪胺酸酶的抑制活性試驗發現這些異黃酮化合物皆具有抑制酪胺酸酶的效果，尤其是以7,8-二羥基異黃酮類化合物(1-4)對酪胺酸酶的抑制作用最佳，其中又以7,8-Dihydroxy-4''-methoxy- isoflavone (2) 和7,8-Dihydroxy- 3''- methoxyisoflavone (3) 抑制作用最佳，在30鐘及60分鐘的IC50值分別為IC50= 50.88 ± 1.80 μM及IC50= 51.83 ± 4.61 μM和IC50= 53.87 ± 11.91 μM及IC50= 54.20 ± 9.21 μM，具有顯著的酪胺酸酶抑制活性為熊果素的3倍 (IC50= 154.02 ± 6.32 μM 及IC50= 146.20 ± 8.32 μM)。除此之外，這些異黃酮類化合物於小鼠黑色素瘤細胞株（B16F10）的細胞存活率試驗，經24小時培養後顯示為具有較低的細胞毒性，最小的細胞存活率為60.38 ± 1.23%，出現在化合物3於濃度80μM下，其餘皆有75%以上的細胞存活率。
本研究結果發現結構中合併有A環具鄰位二羥基與親脂性B環的異黃酮化合物，具有不錯的的酪胺酸酶抑制活性。因此7,8-二羥基異黃酮類化合物是有潛力可作為酪胺酸酶的抑制劑。

Eight isoflavones were synthesized by one-pot approach and tested to find out the inhibiting effect to mushroom tyrosinase and B16F10 cell viability .
These isoflavones were 7,8-dihydroxylated and 5,7-dihydroxylated series based on the ring A system with various ring B substitution. 7,8- dihydroxylated series includes 7,8-Dihydroxyisoflavone (1), 7,8-Dihydroxy- 4''-methoxyisoflavone (2), 7,8-Dihydroxy-3''-methoxyisoflavone (3) and 3'',7,8-Trihydroxyisoflavone (4). 5,7-dihydroxylated series includes 5,7-Dihydroxyisoflavone (5), 5,7-Dihydroxy-4''-methoxyisoflavone (6), 5,7-Dihydroxy-3''-methoxyisoflavone (7), and 3'',5,7-Trihydroxyisoflavone (8). All the structures of compounds were identified by spectroscopic and confirmed by comparing with literature data.
The results showed that isoflavones are effective mushroom tyrosinase inhibitors, especially the 7,8-dihydroxylated series (1-4) has the best inhibitions. Among the isoflavones, the potency of IC50 in 7,8-Dihydroxy-4''-methoxyisoflavone (2) at 30min and 60min are
50.88±1.80 μM & 51.83±4.61 μM and IC50 in 7,8-Dihydroxy-3''-methoxyisoflavone (3) for 53.87 ± 11.91 μM and 54.20± 9.21 μM exhibited most notable inhibiting activities, three times in potency of arbutin (IC50=154.02 ± 6.32 μM & IC50=146.20 ± 8.32 μM). In addition, these synthesized isoflavones had lower cytotoxic after they had been cultivated for 24 hours and then tested for the viability in the murine melanoma celllines (B16F10). The minimum cell viability of these isoflavones was found to be 60.38 ± 1.23% treated with 80μM of 3. The others were found to be up to 75%.
The results showed that the presented isoflavones combined with ortho dihydroxylated ring A combined with lipophilic ring B have good tyrosinase inhibition. Therefore, 7,8-dihydroxylated isoflavones could be potential candidates for new ingredients of tyrosinase inhibitors.

中文摘要 I
Abstract II
謝誌 IV
目錄 VI
圖目錄 VIII
表目錄 XI

第一章 序論 1
第一節 前言 1
第二節 文獻回顧 3
第三節 異黃酮化合物的合成方法 19
第二章 研究動機與目的 21
第三章 研究方法 23
第一節 合成步驟 23
第二節 儀器與材料 29
第三節 生物活性試驗篩選方法 31
第四章 結果與討論 35
第一節 化合物製備及結構證明 35
第二節 異黃酮化合物的合成結果與討論 72
第三節 化合物抑制酪胺酸酶活性試驗 74
第五章 結論 81
第六章 化合物之物理及光譜數據整理 82
第七章 參考文獻 88


圖目錄
圖1-2-1 黃酮類的基本結構 3
圖1-2-2 類黃酮化合物的基本結構 4
圖1-2-3 1,2-Diphenylpropane Isoflavonoids 5
圖1-2-4 1,3-Diphenylpropane Flavonoids 5
圖1-2-5 異黃酮類的生合成 6
圖1-2-6 皮膚結構圖 10
圖1-2-7 黑色素生合成圖 12
圖1-2-8 皮膚光老化的作用機轉 14
圖1-3-1 去氧苯因類合成異黃酮類化合物的合成路徑 20
圖3-1-1 7,8-二羥基異黃酮類的合成步驟 24
圖3-1-2 異黃酮類的環化機轉 25
圖3-1-3 5,7-二羥基異黃酮類的合成步驟 27
圖4-1-1-1 7,8-二羥基異黃酮 (1) 之氫光譜圖 37
圖4-1-1-2 7,8-二羥基異黃酮 (1) 之碳光譜圖 38
圖4-1-2-1 7,8-二羥基-4''-甲氧基異黃酮 (2) 之氫光譜圖 41
圖4-1-2-2 7,8-二羥基-4''-甲氧基異黃酮 (2) 之碳光譜圖 42
圖4-1-2-3 7,8-二羥基-4''-甲氧基異黃酮 (2) 之電子游離質譜圖 43
圖4-1-3-1 7,8-二羥基-3''-甲氧基異黃酮 (3) 之氫光譜圖 46
圖4-1-3-2 7,8-二羥基-3''-甲氧基異黃酮 (3) 之碳光譜圖 47
圖4-1-3-3 7,8-二羥基-3''-甲氧基異黃酮 (3) 之電子游離質譜圖 48
圖4-1-4-1 3'',7,8-三羥基異黃酮 (4) 之氫光譜圖 51
圖4-1-4-2 3'',7,8-三羥基異黃酮 (4) 之碳光譜圖 52
圖4-1-4-3 3'',7,8-三羥基異黃酮 (4) 之電子游離質譜圖 53
圖4-1-5-1 5,7-二羥基異黃酮 (5) 之氫光譜圖 56
圖4-1-5-2 5,7-二羥基異黃酮 (5) 之碳光譜圖 57
圖4-1-6-1 5,7-二羥基-4''-甲氧基異黃酮 (6) 之氫光譜圖 60
圖4-1-6-2 5,7-二羥基-4''-甲氧基異黃酮 (6) 之碳光譜圖 61
圖4-1-6-3 5,7-二羥基-4''-甲氧基異黃酮 (6) 之電子游離質譜圖 62
圖4-1-7-1 5,7-二羥基-3''-甲氧基異黃酮 (7) 之氫光譜圖 65
圖4-1-7-2 5,7-二羥基-3''-甲氧基異黃酮 (7) 之碳光譜圖 66
圖4-1-8-1 3'',5,7-三羥基異黃酮 (8) 之氫光譜圖 69
圖4-1-8-2 3'',5,7-三羥基異黃酮 (8) 之碳光譜圖 70
圖4-1-8-3 3'',5,7-三羥基異黃酮 (8) 之電子游離質譜圖 71
圖4-2-1 合成的異黃酮化合物的產物結構 73
圖4-3-1 化合物1-8 對酪胺酸酶IC50的抑制濃度 75
圖4-3-3 異黃酮化合物1-8對酪胺酸酶30分鐘後的抑制活性試驗結果 78
圖4-3-4 異黃酮化合物1-8對酪胺酸酶60分鐘後的抑制活性試驗結果 78
圖4-3-2 化合物1-8對B16F10的細胞存活率試驗結果 80



表目錄
表1-1-1 衛生署公告化粧品之美白成分 2
表1-2-1 市售美白添加劑的作用機轉 18
表4-3-1 異黃酮化合物1-8對酪胺酸酶IC50的抑制濃度 75
表4-3-2 異黃酮化合物1-8對酪胺酸酶抑制活性試驗結果 76
表4-3-3 化合物1-8對B16F10的細胞存活率試驗結果 80
表6-1-1 化合物1-8的碳譜整彙 87

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