臺灣株真菌Alternaria sp. NTOU3914、Microsphaeropsis arundinis NTOU816和Xylaria intraflava YMJ725發酵液成分之研究

Chemical constituents from the fermented broths of Alternaria sp. NTOU3914, Microsphaeropsis arundinis NTOU 816, and Xylaria intraflava YMJ725 isolated in Taiwan

彭子宴(Tzu-Yen Peng)

指導教授 ： 李宗徽

臺北醫學大學/藥學院/生藥學研究所/碩士(2014年)

https://doi.org/10.6831/TMU.2014.00080

摘要

利用抑制肺癌細胞 (A549) 生長及抑制一氧化氮合成酶的活性分析平台來篩選真菌培養株，已篩選出具細胞毒性或抗發炎潛力的真菌株，而後選定Alternaria sp. NTOU3914、 Microsphaeropsis arundinis NTOU816和Xylaria intraflava YMJ725三株進行其成分研究。以麥芽萃取物 (malt extract)、葡萄糖 (dextrose)、蛋白腖 (peptone)、酵母菌抽出物 (yeast extract) 為培養基加以擴大培養，針對發酵液所含代謝產物進行分析、分離與純化，計獲得13個化合物，分別為cyclo 4-hydroxy-L-Pro-L-Leu (1)、cyclo L-Pro-L-Leu (2)、3-methylorsellinic acid (3)、alternariol 9-monomethyl ether(4)、modiolin (5)、ethyl 2Z,4E-dien-6,9-dihydroxyldecanoate (6)、curvulalic acid (7)、2Z,4E-dien-6,9-dihydroxyldecanoic acid (8)、7-hydroxy-2-methyl-4-chromanon (9)、modiolide B (10)、epoxydon (11)、epiepoformin (12) 及4-oxo-4H-pyran-3-acetic acid (13)。在化合物的骨架分類上，化合物1和2為二酮環縮二氨酸類 (diketopiperazine)，化合物3和4為酚酸類 (phenolics)，化合物5-13為聚酮類 (polyketides)，其中6和8為新化合物。在生物活性試驗上，化合物1-3和5-13對於肺癌細胞 (A549) 抑制生長的GI50皆大於30 μM，化合物4的GI50為21.4 μM。化合物11和12對於RAW264.7細胞產生一氧化氮具有較強的抑制效果，其IC50分別為6.07和8.10 ?嵱。

關鍵字

Alternaria sp. ； Microsphaeropsis arundinis ； Xylaria intraflava ； 次級代謝物 ； 抑制生長活性 ； A549 ； RAW264.7

並列摘要

In this study, three fungal strains classified as Alternaria sp. NTOU3914, Microsphaeropsis arundinis NTOU816, and Xylaria intraflava YMJ725, were chosen for chemical investigations of their secondary metabolites on the basis of the preliminary screening. The crude extracts of the fermented broths of Alternaria sp. NTOU3914 and X. intraflava YMJ725 exhibited significant growth inhibitory activities against A549 lung cancer cells, and the crude extract of the fermented broth of M.arundinis NTOU816 exhibited significant anti-iNOS activity in LPS-induced RAW264.7 cells. Thus, a series of separation and purification on the fermented broths of these fungal strains was carried out, which has resulted in the isolation and identification of thirteen compounds 1-13 including together with cyclo 4-hydroxy-L-Pro-L-Leu (1), cyclo L-Pro-L-Leu (2), 3-methylorsellinic acid (3), alternariol 9-monmethyl ether (4), modiolin (5), ethyl 2Z,4E-dien-6,9-dihydroxyldecanoate (6), curvulalic acid (7), 2Z,4E-dien-6,9-dihydroxyldecanoic acid (8), 7-hydroxy-2-methyl-4-chromanon (9), modiolide B (10), epoxydon (11), epiepoformin (12) and 4-oxo-4H-pyran-3-acetic acid (13). Compounds 1-3 and 5-13 exhibited slight growth inhibitory activities against A549 cell line with GI50 values more than 30.0 μM, while 4 exhibited moderate growth inhibitory activity against A549 cell line with a GI50 value of 21.4 μM compared to Fluorouracil (GI50 = 3.6 μM). In addition, compounds 11 and 12 exhibted potent inhibitory activity on NO production of RAW264.7 cells induced by lipopolysaccharide with IC50 values of 6.07, and 8.10 ?嵱, respectively, compared to aminoguanidine and N??-nitro-L-arginine (IC50 = 21.22, 142.37 ?嵱).

並列關鍵字

Alternaria sp. ； Microsphaeropsis arundinis ； Xylaria intraflava, secondary metabolite ； growth inhibitory activity ； A549 ； RAW264.7

參考文獻

66. 李麗雯, 台灣兩種海洋細菌之生物活性成分研究. 台北醫學大學生藥所碩士論文 2007.

1. Berdy, J., Bioactive Microbial Metabolites. J. Antibiot 2005, 58, 1-26.

2. Walsh, J. C. C., Lessons from natural molecules. Nature 2004, 432, 829-837.

3. Newman, D. J.; Cragg, G. M., Natural products as sources of new drugs over the 30 years from 1981 to 2010. J. Nat Prod 2012, 75, 311-35.

4. Evidente, A.; Kornienko, A.; Cimmino, A.; Andolfi, A.; Lefranc, F.; Mathieu, V.; Kiss, R., Fungal metabolites with anticancer activity. Nat Prod Rep 2014, 31, 617-27.