臺灣博碩士論文加值系統

從採集自小琉球海域的巨大鞘絲藻Lyngbya majuscula 中，分離純化出八個化合物，包含四個malyngamides 類的化合物(1 ~ 4)，其中malyngamide Z (1)、malyngamide X (2) 及malyngamide 1 (4) 為新化合物，isomalyngamide A (3) 為已知化合物；四個aplysiatoxins 類的化合物(5 ~ 7)，其中3-methoxy debromoaplysiatoxin (7)為新化合物，anhydrodebromoaplysiatoxin (5)、debromoaplysiatoxin (6) 及oscillatoxin B (8)為已知化合物，上述化合物的結構皆以核磁共振圖譜以及文獻 資料比對後加以確認。
在癌細胞毒殺活性測試中，化合物1、3 及化合物5、6、7、8對於人類肝癌細胞 (Hep2) 與人類大腸直腸癌細胞 (WiDr) 均具有顯著的抑制活性，ED50 介於11.6~26.0 µg/ml，在豐年蝦毒殺試驗中，malyngamides 類的化合物(1、3、4)沒有顯著毒殺活性，而aplysiatoxins類的化合物(5 ~ 8)則皆具有顯著的豐年蝦毒殺活性，其LD50 介於0.5 ~ 7.7 µg/ml。

In continuing search the bioactive secondary metabolites from marine microorganism, we found that the extracts of marine cyanobacteria Lyngbya majuscula, collected from Ryukyu Island, showed significant anticancer and anti-dialetes activities. Through bioassay-guided separation, we isolated four malyngamides, including three new compounds, malyngamide Z (1), malyngamide X (2), malyngamide 1 (4) and a known compound, isomalyngamide A (3). We also isolated four aplysiatoxins, including one new compound, 3-methoxy debromoaplysiatoxin (7) and three known compounds, ahhydrodebromoaplysiatoxin (6), debromoaplysiatoxin and oscillatoxin B (8) from L. majuscula. The structure of these compounds were determined by 1D and 2D NMR spectroscopic analysis.
In the biological test, compound 1、3 and 5-8 exhibited activity in cell lines of human hepatoma 2 (Hep2) and human colorectal cancer (WiDr). The ED50 values were between 11.6 ~ 26.0µg/ml. In brine shrimp toxicity test, malyngamides (1、3、4) showed no activity and aplysiatoxins 5 ~ 8 showed strong activity. The LD50 values were between 0.5 ~ 7.7 µg/ml.

壹、緒論..................................................1

1.1 前言..................................................1

1.2 研究動機.............................................20

貳、實驗結果與討論.......................................21

2.1 萃取與分離純化步驟...................................21

2.2 Malyngamide X (1)之結構解析..........................23

2.3 Malyngamide Z (2)之結構解析..........................32

2.4 Isomalyngamide A (3) 之結構解析......................40

2.5 Malyngamide 1 (4) 之結構解析.........................49

2.6 Anhydrodebromoaplysiatoxin (5) 之結構
解析.................................................58

2.7 Debromoaplysiatoxin (6) 之結構解析...................66

2.8 3-methoxy aplysiatoxin (7) 之結構解析................74

2.9 Oscillatoxin B (8) 之結構解析........................82

2.10 生物活性............................................90

2.10.1 癌細胞毒殺分析實驗................................90

2.10.2 豐年蝦毒殺活性實驗................................91

2.11 結論................................................92

參、實驗部分.............................................93

3.1 儀器與高效能液相層析系統.............................93

3.2 巨大鞘絲藻Lyngbya majuscula 之粗萃與分離流程.........94

3.3 正己烷層的純化.......................................95

3.4 二氯甲烷層的純化.....................................96

3.5 癌細胞毒殺活性.......................................98

3.6 豐年蝦毒殺活性實驗...................................99

3.7 實驗數據整理........................................100

肆、參考文獻............................................103

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