2019-01-012024-05-16https://scholars.lib.ntu.edu.tw/handle/123456789/667584摘要：中文摘要： 高科技除了帶來福祉，也造成嚴重的環境污染，人們所面臨的威脅是每ㄧ年癌症的死亡率居高不下，復由於藥物濫用導致病菌產生抗藥性越來越嚴重的情形下，臨床使用抗生素的藥效呈每況愈下的趨勢，且已演變為全球普遍存在的問題，解決之道除了減少環境的污染外，如何開發出強效低負作用的新藥，仍刻不容緩。台灣位處熱帶、亞熱帶，適合藻類的生長，特別是東北角海岸線長達67公里，屬於亞熱帶氣候，海域有兩股洋流在此交會，冬季以大陸沿岸流的冷水團為主，夏季則以黑潮的暖水團為主，富含營養鹽，因此孕育了高度多樣性的大型藻種，目前共發現200多種，已鑑定者有179種，多年來已有相當多本土學者投入大型藻的天然物研究，並陸續有新穎的化合物被報導出來，然而關於藻類的衍生真菌的天然物相關研究目前仍屈指可數。本計畫規劃在未來三年內，擬以東北角海岸常見的大型藻之衍生真菌株為研究對象，在單離、純化獲得真菌株後，初步以抗菌和抗癌為篩選平臺，經篩得具活性的菌株後，再進行後續的發酵實驗和天然物的研究，茲規劃如下： 第一年：單離藻源真菌株，預計至少1500株，以瓊脂培養基皿培 (agar plate) 後，進行抗菌初步篩選，挑選出活性較佳的菌株亦進行癌細胞毒性篩選，其中具顯著活性之菌株進行形態分類與18S rDNA鑑種，萃出物以HPLC進行micro-fractionation，再以UPLC/MS/MS分析，搭配資料庫進行dereplication，其中具研究潛力的菌株進行後續天然物研究。 第二年：經挑選會產出活性新穎化合物的菌株，以OSMAC的模式進行培養基測試與擴量培養，再以HPLC進行不同發酵萃出物成分差異分析，針對具活性的波峰，以活性導引 (bioassay-guided) 的方式進行分離、純化與構造解析，並進行純質之深入藥理評估。 第三年：除延續第二年的工作外，針對具活性的新穎化合物，利用改變培養方式獲得一系列類似物，或針對含量多的化合物進行結構修飾，搭配活性平台，進行構效 (structure-activity relationship) 分析與深入的藥理研究，包括：抗菌試驗、抗癌試驗、抗發炎試驗與抗血管增生試驗等，期能開發出具應用潛力的先導化合物。 <br> Abstract: The high-tech has improved the human welfare, and which also cause serious pollution simultaneously. The threats of the annual death caused by cancer are remaining high. The drug resistance of the pathogen is getting more serious due to the drug abuse, which results in the efficacy of the antibiotics being worse and worse. This problem has been developed as a global issue, and reducing the pollution and developing effective new drugs with no side effect seem to be of great urgency. Taiwan is located at tropical and subtropical region, where it is quite suitable for the growth of macroalgae. Especially, there are 67 kilometer along the north-east coast line of Taiwan, and the convergence of two ocean currents make the nutrients more abundant. The species diversity of local macroalgae is high, and of the over 200 species found, 179 species have been identified. The natural product research of the macroalgae has been performed for many years, and many novel compounds were reported successively. However, the chemical investigation of macroalgae-derived fungal strains remains rare. Thus, within three years this project is designed to focus on the natural product research of the macroalgae-derived fungal strains. After the fungal strains being isolated and purified, all the strains are subjected to screening using antimicrobial and anticancer platforms. Then, the active strains are mass cultured and studied on their active principles. The detail planning of the three years project is drawn up as below. In the first year, at least 1,500 fungal strains will be isolated from macroalgae. The crude extracts of the petri dish-cultured fungal strains will be subjected to antimicrobial and anticancer screening. Of all the strains screened, the bioactive strains will be identified by morphology and 18S rDNA. HPLC microfractionation coupled with UPLC/MS/MS will be performed for further dereplication. In the second year, the selected fungal strains that could produce novel compounds will be mass cultured. The media used for mass culture will be developed by an OSMAC method using different combination of the authentic nutrients. The bioactive compounds in the fermented products will be purified by a bioassay-guided method. All the obtained compounds will be identified by spectral analysis and subjected to pharmacological studies intensively. In the third year, in addition to follow the works of the 2nd year more chemical analogues of the bioactive compounds will be afforded by changing the culturing conditions of the fungal strains. Bioactive compounds with large amount will be modified for further structure-activity relationship studies. Moreover, all the obtained compounds will be subjected to pharmacological studies including antimicrobial, anti-inflammatory and antiangiogenesis. Hopefully, some leading drugs can be developed from the isolated fungal strains.海藻內生真菌抗癌抗菌新藥開發marine algaeendophyteanticancerantimicrobialnew drug development