【作者】 陈娟；

【导师】 顾谦群； 朱伟明；

【作者基本信息】 中国海洋大学 ， 药物化学， 2006， 硕士

【摘要】 海绵共附生微生物因其次级代谢产物结构新颖、活性独特,目前已成为抗肿瘤药物研究的重要资源。本文对两株海绵来源的抗肿瘤活性真菌黄灰青霉Penicillium auratiogriseum(Sp-19)和菌株CTFS-15的抗肿瘤活性次级代谢产物进行了较系统地研究。此外,本文还对利用宏基因技术从未培养海绵共附生微生物中寻找生物活性代谢产物的方法进行了初步探索。对黄灰青霉P. auratiogriseum(Sp-19)和菌株CTFS-15分别进行大量发酵,发酵液经乙酸乙酯萃取得粗提物;采用人慢性髓性白血病K562细胞,以细胞增殖抑制活性结果为分离向导,采用减压硅胶柱层析、凝胶柱层析(Sephadex LH-20)、反相硅胶柱层析(RP-18)和反相制备高效液相(HPLC)等分离手段,从两株真菌的乙酸乙酯提取物中分离得到27个化合物;利用理化性质和光谱学方法(UV、IR、MS、1D-NMR和2D-NMR)鉴定了其中23个化合物。其中分离自黄灰青霉P. auratiogriseum的18个化合物分别为:Auratiomide A(1),Auratiomide C(2),Anacine(3),Fructigenines A(4),Fructigenines B(5),Pseurotin A(6),青霉菌素(7),青霉菌醇(8),Viridicatol(9),3-甲氧基-4-苯基喹啉酮(10),3-甲基喹唑啉酮(11),(2S,4S)-2,4,8-trihydroxy-3,4-dihydronaphthalen-1(2 H)-one(12),(2S,4R)-2,4,8-trihydroxy-3,4-dihydronaphthalen-1(2H)-one(13),(3R,4R)-3,4,8-trihydroxy-3,4-dihydronaphthalen-1(2H)-one(14),麦角甾醇(15),N-(2-羟苯基)乙酰胺(16),N-(2-(甲氨基)苯基)乙酰胺(17)和大黄素(18);分离自菌株CTFS-15的5个化合物分别为:过氧化麦角甾醇(19),对羟基苯乙酰胺(20),7-羟基-4-甲氧基-5-甲基香豆素(21),环-(丙-亮)(22),靛红(23)。利用人(K562、A549、BEL-7402、HL-60)及小鼠(tsFT210、P388)肿瘤细胞,以细胞增殖抑制率、细胞凋亡诱导和细胞周期抑制活性为指标,应用流式细胞术、SRB等方法对分离得到的化合物进行抗肿瘤活性的初步评价。结果表更多还原

【Abstract】 Sponge-derived microorganisms have become an important source for researches on antitumor drugs, because they can produce structurally novel and biologically active secondary metabolites. This work describes the study on the secondary metabolites of two sponge-derived antitumor active fungi, identified as Penicillium auratiogriseum (Sp-19) and unknown strain CTFS-15. The isolated compounds were evaluated for their antitumor activities. Besides, the work also describes the study on the approach to explore bioactive metabolites of sponge-derived uncultured symbionts through the genetic engineering technology.Two active fungi strains were fermented, respectively. The whole broth was extracted with ethyl acetate to give ferment extracts. Monitoring by the bioassay-guided isolation, the extracts were separated by means of modern chromatographic methods including silica gel column chromatography, Sephadex LH-20, RP-18 and reverse HPLC. Twenty seven compounds were isolated and twenty three of them were elucidated for structures by spectroscopic methods (UV、IR、MS、1D-NMR and 2D-NMR) and physicochemical properties. Compounds isolated from P. auratiogriseum are Auratiomide A(1), Auratiomide C(2), Anacine(3), Fructigenines A(4), Fructigenines B(5), Pseurotin A(6), Cyclopenin(7), Cyclopenol(8), 3-hydroxy-4 -(3-hydroxyphenyl)quinolin-2(1H)-one(9), O-Methylviridicatin(10), 3-methylquinazo lin-4(3H)-one(11), (2S,4S)-2,4,8-trihydroxy-3,4-dihydronaphthalen-1(2H)-one(12), (2S,4R)-2,4,8-trihydroxy-3,4-dihydronaphthalen-1(2H)-one(13), (3R,4R)-3,4,8-trihyd roxy-3,4-dihydronaphthalen-1(2H)-one(14), Ergosterol(15), N-(2-hydroxyphenyl)acet amide(16), N-(2-(methylamino)phenyl)acetamide(17) and Emodin(18). Compounds更多还原