粗毛纤孔菌与椭圆嗜蓝孢孔菌子实体的化学成分及其药理活性研究
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摘要
本文以天然产物化学和药理学理论为基础,对两种药用木腐真菌粗毛纤孔菌(Inonotus hispidus)和椭圆嗜蓝孢孔菌(Fomitiporia ellipsoidea)的化学成分进行提取分离、结构鉴定和药理活性研究。通过硅胶、ODS、Sephadex LH-20、pre-HPLC等各种色谱分离手段,借助1H-NMR、13C-NMR、1H-1H COSY、HMQC、HMBC等现代波谱学测试分析方法,并结合EI-MS、ESI-MS和HR-ESI-MS等质谱技术,共分离出28个化合物,鉴定出27个其中包括4个新化合物,分别命名为Inonotusin A和Inonotusin B、Fomitiporiaester A和Phelligridin K,新化合物类型为吡喃酮类多酚化合物及呋喃酯类物质。并对分离得到的化合物和提取物进行了体内外抗肿瘤活性、体外抗氧化和抑菌活性评价。
粗毛纤孔菌在新疆南部主要寄生在40年以上的古老桑树上,南疆维吾尔族人民有以其作为“桑黄”采集入药的习惯,主要用于治疗各种癌症、糖尿病、痛风和关节炎等疑难疾病。本文从该菌子实体中分离并鉴定出15个化合物,包括7(8),22(23)-二烯-3-酮-麦角甾烷、麦角甾醇、(22E,24R)-5α,8α-过氧麦角甾-6,22-二烯-3p-醇、4,6,8(14),22(23)-四烯-3-酮-麦角甾烷、齿孔酸5个麦角甾类物质；Hispidin、Hispolon、Inonotusin A、InonotusinB、4-(3,4-二羟苯基)-3-丁烯-2-酮、Inoscavin C、Inoscavin D及原儿茶酸和原儿茶醛9个多酚类物质；1个次黄嘌呤核苷物质。并运用SRB法对分离得到的化合物Inonotusin A、 Inonotusin B、4-(3,4-二羟苯基)-3-丁烯-2-酮、Hispidin及原儿茶醛的体外细胞毒活性进行测定,结果仅Inonotusin A对乳腺癌细胞(MCF-7)有适中的活性,IC50值为19.6μM,而其他4个化合物对乳腺癌细胞(MCF-7)和肝癌细胞(HepG-2)均没有明显活性。通过H22荷瘤小鼠移植模型对该菌不同成熟期子实体的石油醚提取物和水提取物的体内抗肿瘤活性进行评价,结果成熟期石油醚低剂量(50mg/Kg)和高剂量(100mg/Kg)抑瘤率分别为44.40%和55.24%,成熟期水提层低剂量(500mg/Kg)和高剂量(1000mg/Kg)抑瘤率分别为54.01%和52.19%,与阳性药CTX接近(61.36%),晚期(子实体变黑)石油醚提取物抑瘤率明显低于成熟期石油醚提取物,低剂量(50mg/Kg)抑瘤率为34.23%,而高剂量则没有活性；晚期甲醇低剂量组抑瘤率为29.77%,水提物低剂量为29.99%,高剂量没有明显活性,晚期水提物抑瘤活性远低于成熟期水提物抑瘤活性。水层提取物荷瘤小鼠白细胞数明显高于环磷酰胺组及石油醚提取物组,脾指数和胸腺指数均有一定程度的提高,联合用药组脾指数和胸腺指数及体重增加均高十阳性CTX组。石油醚组小鼠体重增加低于其他给药组。另外,采用福林酚法和NaNO2-Al(NO3)3比色法分别测定了粗毛纤孔菌子实体70%乙醇提取物的多酚含量和黄酮含量,含量分别为64.02±0.73μg/mg和5.74±0.13μg/mg；并对70%乙醇提取物及主要化合物Hispidin的抗氧化和抑菌活性进行了测定,结果70%乙醇提取物和Hispidin具有强烈的清除DPPH和·OH自由基的活性,在浓度为200mg/Kg时与BHA相当,清除率达到90%左右：采用ABTS试剂盒法测定了化合物Inonotusin A、Inonotusin B、4-(3,4-二羟基苯基)-3-丁烯-2-酮、Hispidin和原儿茶醛5种化合物的抗氧化活性,结果Inonotusin B表现出强烈的清除ABTS自由基的活性,达到59.53±9.70μmol Trolox。采用K-B纸片扩散法测定了70%乙醇提取物及Hispidin对大肠杆菌Escherichia coli ATCC9099、金黄色葡萄球菌Staphyloccocus aureus ATCC6538、枯草芽孢杆菌Bacillus subtilis的抑制活性,结果除70%乙醇提取物高剂量(40.96mg/mL)对大肠杆菌的抑制率达到40.35%,其余均没有明显活性。
椭圆嗜蓝孢孔菌是崔宝凯和戴玉成研究员于2008年首次在福建省建瓯市万亩林自然保护区发现,该地区民间俗称桑黄,2010年戴玉成研究员再次在中国海南岛发现该菌,并发现其是目前世界上最大的多孔菌。本文从该菌子实体中分离出13个化合物,鉴定出12个化合物,包括1个呋喃酯类Fomitiporiaester A和11个多酚类物质4-(3,4-二羟苯基)-3-丁烯-2-酮、Hispidin、Hispolon、Inoscavin A、Inoscavin C、Methylinoscavin D、Inoscavin E、 Phelligridin K、Inonoblin B及原儿茶酸和原儿茶醛。并运用SRB法对分离得到的化合物Fomitiporiaester A、Inoscavin C和Inonoblin B的细胞毒活性进行测定,结果Inoscavin C和Inonoblin B对HepG2和Skov3细胞增殖有一定的抑制作用,当浓度为200μM时,InoscavinC对HepG2和Skov3细胞的抑制率分别为77.16%和65.87%；Inonoblins B对两种细胞的最高抑制率分别为57.86%和45.23%,而Fomitiporiaester A对两种癌细胞均无明显细胞毒活性。通过H22荷瘤小鼠移植模型对该菌中分离得到化合物Fomitiporiaester A的体内抗肿瘤活性进行评价,结果该化合物对肝癌H22瘤细胞具有良好的抑制活性,当剂量为5mg/Kg时抑制率为42.94%,在给药量为10mg/Kg时抑制率(49.17%)与阳性药CTX接近(52.56%),当剂量为20mg/Kg时抑制率(58.15%)高于阳性药CTX,抑瘤效果具有-定的量效关系,并和阴性对照组具有显著性差异(p<0.05)。该化合物中高剂量给药组荷瘤小鼠的脾指数和胸腺指数均高于环磷酰胺组,说明该药物毒性低于环磷酰胺。给药组荷瘤小鼠白介素2与阳性组没有明显区别,通过HE、Bax和Bcl-2染色分析可以看出化合物Fomitiporiaester A对荷瘤小鼠肿瘤细胞凋亡有促进作用。另外,通过ABTS试剂盒法测定了椭圆嗜蓝孢孔菌子实体中部分化合物的抗氧化活性,结果显示新化合物FomitiporiaesterA不具备清除ABTS自由基的活性；而其他Hispidin衍生物均具有一定的清除ABTS自由基活性,特别是新化合物Phelligridin K的活性最高,为3.56mM Trolox。
本论文对粗毛纤孔菌和椭圆嗜蓝孢孔菌的化学成分和药理活性进行了研究,分离得到了抗肿瘤和抗氧化活性物质,包括4个新的化合物,为活性先导化合物的发现奠定了基础,同时为粗毛纤孔菌和椭圆嗜蓝孢孔菌两种“桑黄”真菌的开发利用提供了科学依据。
Based on the theories of natural product chemistry and pharmacology, the chemical constituents and bioactivities of Inonotus hispidus and Fomitiporia ellipsoidea were studied. Twenty eight compounds, including four new ones, have been isolated from the fruiting bodies of two medicinal fungi by variety of chromatographic methods (silica gel, reversed-phase (ODS), sephadex LH-20and pre-HPLC) and structures of twenty seven compounds were elucidated on the basis of extensive spectroscopic analysis including1D and2D-NMR(1H-NMR,13C-NMR,1H-1H COSY, HMQC and HMBC) as well as mass spectrometry (EI-MS, ESI-MS and HR-ESI-MS). The new compounds involved pyrone and furfuran ester, and named inonotusin A, inonotusin B, fomitiporiaester A and phelligridin K. Some of the isolates and extracts were evaluated for their vitro and vivo antitumor, antioxidant and antibacterial activities.
Ⅰ. hispidus is a parasitic fungus preferably living on mulberry of more than40years in the southern region of Xinjiang province, and has been used as traditional medicines for the treatment of cancer, diabetes, arthritides and other stomach problems. Fifteen compounds were isolated from the fruiting body of Ⅰ. hispidus, including7(8),22(23)-diene-3-one-ergostane, ergosterol,5α,8α-epidioxy-(22E,24R)-ergosta-6,22-dien-3β-ol,4,6,8(14),22(23)-tetraen-3-one-ergostane, eburicoic acid, hispidin, hispolon, inonotusin A, inonotusin B,(E)-4-(3,4-dihydroxyphenyl) but-3-en-2-one, inoscavin C, inoscavin D, protocatechuic acid, protocatechualdehyde and inosine. Compounds inonotusin A and B,(E)-4-(3,4-dihydroxyphenyl) but-3-en-2-one, hispidin and protocatechualdehyde were tested for cytotoxic activity against two cell lines, human hepatoblastoma(HepG-2) and human breast carcinoma(MCF-7) by SRB method. The result showed only inonotusin A had moderate cytotoxicity against MCF-7with IC50values of19.6μM, and other four compounds were noncytotoxic to the two cell lines. In vivo anti-tumor activity of petroleum ether and water extracts from fruiting bodies in different stage also carried out by H22tumor-bearing mice transplanting model. The result showed that petroleum ether and water extracts in maturation stage had significant antitumor activities, and the inhibitory value were44.40%(petroleum ether low dosage group,50mg/Kg),55.24%(petroleum ether high dosage group,100mg/Kg),54.01%(water low dosage group,500mg/Kg),52.19%(water high dosage group,1000mg/Kg), and closed to the inhibition of CTX group (61.36%). The water extracts could regulate the immune function of tumor-bearing mice, but petroleum ether extract had effection to immune organ negatively. In darkening fruiting body stage, the inhibitory effects of petroleum ether and water extracts were more lower than the maturation stage, and the value were34.73%(50mg/Kg) and29.99%(500mg/Kg), but the high dosage group had no activity to H22. The inhibition value of methanol extracts was29.77%(500mg/Kg). The extracts had the similar effects to immune organ of tumor-bearing mice, the water extracts could regulate the immune function, but petroleum ether extracts had effection to immune organ negatively. The leucocyte, spleen index and thymus index of water extracts increased higher than CTX group. Moreover, the concentrations of phenolic compounds in the70%ethanol extract of Inonotus hispidus were measured with Folin-Ciocalteu reagent, and total flavonoid concentration was determined by the NaNO2-Al(NO3)3method. The result showed the phenolics were64.02±0.73μg/mg(gallic acid) while total flavanoids were5.74±0.13μg/mg (rutin equivalent) in the70%extract. Antioxidant and antimicrobial activities of Ⅰ. hispidus extracts obtained with70%ethanol and compound hispidin were investigated in this study.·DPPH and·OH free radical-scavenging activity were found to90%inhibition at concentration of200μg/mL. Compound inonotusin B potently scavenged the ABTS radical cation and reached to the level of59.53±9.70μmol.70%ethanol extract and hispidin showed narrow antibacterial activities against Staphyloccocus aureus and Bacillus subtilis.70%ethanol extracts inhibited the growth of the Escherichia coli, and inhibition values was40.35%at concentration of40.96mg/mL.
Fomitiporia ellipsoidea was first collected in Wanmulin Nature Reserve in2008by Bao-Kai Cui and Yu-Cheng Dai, and local people called "sanghuang". In2010, Cui and Dai were performing field work in tropical woodland on Hainan Island, China, studying wood-rotting fungi. The pair uncovered a very large F. ellipsoidea fruit body on a fallen Quercus asymetrica log, which turned out to be the largest fungal fruit body ever documented. Thirteen compounds were isolated from the fruiting body of F. ellipsoidea, and twelve compounds were elucidated, including fomitiporiaester A,(E)-4-(3,4-dihydroxyphenyl) but-3-en-2-one, hispidin, hispolon, inoscavin A, inoscavin C, methylinoscavin D, inoscavin E, phelligridin K, inonoblins B, protocatechuic acid and protocatechualdehyde. Compounds fomitiporiaester A、inoscavin C and inonoblins B were tested for cytotoxic activity against two cell lines, human hepatoblastoma(HepG-2) and human oophoroma (SKOV3) by SRB method. The result showed that inoscavin C and inonoblins B had moderate cytotoxicity against the two cell lines and fomitiporiaester A was noncytotoxic to the two cell lines. In vivo anti-tumor activity of fomitiporiaester A was carried out by H22tumor-bearing mice transplanting model. The result indicated fomitiporiaester A had manifest antitumor activity, and the inhibition ration were 42.94%,49.17%and58.15%at concentration of5mg/Kg,10mg/Kg and20mg/Kg, respectively. Inhibition value nearly to the CTX group (52.56%) at dosage10mg/Kg and better than the CTX group at dosage10mg/Kg. Inhibition effects had dose-effect relationship, and it had significant difference to compare negative control (p<0.01), In addition,compound can increase the immune organ index. Some of isolated hispidin derivates from fruiting bady of F. ellipsoidea potently scavenged the ABTS radical cation in which new compound phelligridin K showed the highest activity at the level of3.56mM Trolox. But fomitiporiaester A had no activity to scavenge the ABTS radical cation.
As a conclusion, the chemical constituents and bioactivities of Ⅰ. hispidus and F. ellipsoidea were studied in this thesis. We isolated some compounds with anti-tumor and antioxidant activitives, including four new compounds.The present results established the basis for discovery of bioactive lead compounds and provided theoretical foundation for further utilization of the two species fungi.
粗毛纤孔菌在新疆南部主要寄生在40年以上的古老桑树上,南疆维吾尔族人民有以其作为“桑黄”采集入药的习惯,主要用于治疗各种癌症、糖尿病、痛风和关节炎等疑难疾病。本文从该菌子实体中分离并鉴定出15个化合物,包括7(8),22(23)-二烯-3-酮-麦角甾烷、麦角甾醇、(22E,24R)-5α,8α-过氧麦角甾-6,22-二烯-3p-醇、4,6,8(14),22(23)-四烯-3-酮-麦角甾烷、齿孔酸5个麦角甾类物质；Hispidin、Hispolon、Inonotusin A、InonotusinB、4-(3,4-二羟苯基)-3-丁烯-2-酮、Inoscavin C、Inoscavin D及原儿茶酸和原儿茶醛9个多酚类物质；1个次黄嘌呤核苷物质。并运用SRB法对分离得到的化合物Inonotusin A、 Inonotusin B、4-(3,4-二羟苯基)-3-丁烯-2-酮、Hispidin及原儿茶醛的体外细胞毒活性进行测定,结果仅Inonotusin A对乳腺癌细胞(MCF-7)有适中的活性,IC50值为19.6μM,而其他4个化合物对乳腺癌细胞(MCF-7)和肝癌细胞(HepG-2)均没有明显活性。通过H22荷瘤小鼠移植模型对该菌不同成熟期子实体的石油醚提取物和水提取物的体内抗肿瘤活性进行评价,结果成熟期石油醚低剂量(50mg/Kg)和高剂量(100mg/Kg)抑瘤率分别为44.40%和55.24%,成熟期水提层低剂量(500mg/Kg)和高剂量(1000mg/Kg)抑瘤率分别为54.01%和52.19%,与阳性药CTX接近(61.36%),晚期(子实体变黑)石油醚提取物抑瘤率明显低于成熟期石油醚提取物,低剂量(50mg/Kg)抑瘤率为34.23%,而高剂量则没有活性；晚期甲醇低剂量组抑瘤率为29.77%,水提物低剂量为29.99%,高剂量没有明显活性,晚期水提物抑瘤活性远低于成熟期水提物抑瘤活性。水层提取物荷瘤小鼠白细胞数明显高于环磷酰胺组及石油醚提取物组,脾指数和胸腺指数均有一定程度的提高,联合用药组脾指数和胸腺指数及体重增加均高十阳性CTX组。石油醚组小鼠体重增加低于其他给药组。另外,采用福林酚法和NaNO2-Al(NO3)3比色法分别测定了粗毛纤孔菌子实体70%乙醇提取物的多酚含量和黄酮含量,含量分别为64.02±0.73μg/mg和5.74±0.13μg/mg；并对70%乙醇提取物及主要化合物Hispidin的抗氧化和抑菌活性进行了测定,结果70%乙醇提取物和Hispidin具有强烈的清除DPPH和·OH自由基的活性,在浓度为200mg/Kg时与BHA相当,清除率达到90%左右：采用ABTS试剂盒法测定了化合物Inonotusin A、Inonotusin B、4-(3,4-二羟基苯基)-3-丁烯-2-酮、Hispidin和原儿茶醛5种化合物的抗氧化活性,结果Inonotusin B表现出强烈的清除ABTS自由基的活性,达到59.53±9.70μmol Trolox。采用K-B纸片扩散法测定了70%乙醇提取物及Hispidin对大肠杆菌Escherichia coli ATCC9099、金黄色葡萄球菌Staphyloccocus aureus ATCC6538、枯草芽孢杆菌Bacillus subtilis的抑制活性,结果除70%乙醇提取物高剂量(40.96mg/mL)对大肠杆菌的抑制率达到40.35%,其余均没有明显活性。
椭圆嗜蓝孢孔菌是崔宝凯和戴玉成研究员于2008年首次在福建省建瓯市万亩林自然保护区发现,该地区民间俗称桑黄,2010年戴玉成研究员再次在中国海南岛发现该菌,并发现其是目前世界上最大的多孔菌。本文从该菌子实体中分离出13个化合物,鉴定出12个化合物,包括1个呋喃酯类Fomitiporiaester A和11个多酚类物质4-(3,4-二羟苯基)-3-丁烯-2-酮、Hispidin、Hispolon、Inoscavin A、Inoscavin C、Methylinoscavin D、Inoscavin E、 Phelligridin K、Inonoblin B及原儿茶酸和原儿茶醛。并运用SRB法对分离得到的化合物Fomitiporiaester A、Inoscavin C和Inonoblin B的细胞毒活性进行测定,结果Inoscavin C和Inonoblin B对HepG2和Skov3细胞增殖有一定的抑制作用,当浓度为200μM时,InoscavinC对HepG2和Skov3细胞的抑制率分别为77.16%和65.87%；Inonoblins B对两种细胞的最高抑制率分别为57.86%和45.23%,而Fomitiporiaester A对两种癌细胞均无明显细胞毒活性。通过H22荷瘤小鼠移植模型对该菌中分离得到化合物Fomitiporiaester A的体内抗肿瘤活性进行评价,结果该化合物对肝癌H22瘤细胞具有良好的抑制活性,当剂量为5mg/Kg时抑制率为42.94%,在给药量为10mg/Kg时抑制率(49.17%)与阳性药CTX接近(52.56%),当剂量为20mg/Kg时抑制率(58.15%)高于阳性药CTX,抑瘤效果具有-定的量效关系,并和阴性对照组具有显著性差异(p<0.05)。该化合物中高剂量给药组荷瘤小鼠的脾指数和胸腺指数均高于环磷酰胺组,说明该药物毒性低于环磷酰胺。给药组荷瘤小鼠白介素2与阳性组没有明显区别,通过HE、Bax和Bcl-2染色分析可以看出化合物Fomitiporiaester A对荷瘤小鼠肿瘤细胞凋亡有促进作用。另外,通过ABTS试剂盒法测定了椭圆嗜蓝孢孔菌子实体中部分化合物的抗氧化活性,结果显示新化合物FomitiporiaesterA不具备清除ABTS自由基的活性；而其他Hispidin衍生物均具有一定的清除ABTS自由基活性,特别是新化合物Phelligridin K的活性最高,为3.56mM Trolox。
本论文对粗毛纤孔菌和椭圆嗜蓝孢孔菌的化学成分和药理活性进行了研究,分离得到了抗肿瘤和抗氧化活性物质,包括4个新的化合物,为活性先导化合物的发现奠定了基础,同时为粗毛纤孔菌和椭圆嗜蓝孢孔菌两种“桑黄”真菌的开发利用提供了科学依据。
Based on the theories of natural product chemistry and pharmacology, the chemical constituents and bioactivities of Inonotus hispidus and Fomitiporia ellipsoidea were studied. Twenty eight compounds, including four new ones, have been isolated from the fruiting bodies of two medicinal fungi by variety of chromatographic methods (silica gel, reversed-phase (ODS), sephadex LH-20and pre-HPLC) and structures of twenty seven compounds were elucidated on the basis of extensive spectroscopic analysis including1D and2D-NMR(1H-NMR,13C-NMR,1H-1H COSY, HMQC and HMBC) as well as mass spectrometry (EI-MS, ESI-MS and HR-ESI-MS). The new compounds involved pyrone and furfuran ester, and named inonotusin A, inonotusin B, fomitiporiaester A and phelligridin K. Some of the isolates and extracts were evaluated for their vitro and vivo antitumor, antioxidant and antibacterial activities.
Ⅰ. hispidus is a parasitic fungus preferably living on mulberry of more than40years in the southern region of Xinjiang province, and has been used as traditional medicines for the treatment of cancer, diabetes, arthritides and other stomach problems. Fifteen compounds were isolated from the fruiting body of Ⅰ. hispidus, including7(8),22(23)-diene-3-one-ergostane, ergosterol,5α,8α-epidioxy-(22E,24R)-ergosta-6,22-dien-3β-ol,4,6,8(14),22(23)-tetraen-3-one-ergostane, eburicoic acid, hispidin, hispolon, inonotusin A, inonotusin B,(E)-4-(3,4-dihydroxyphenyl) but-3-en-2-one, inoscavin C, inoscavin D, protocatechuic acid, protocatechualdehyde and inosine. Compounds inonotusin A and B,(E)-4-(3,4-dihydroxyphenyl) but-3-en-2-one, hispidin and protocatechualdehyde were tested for cytotoxic activity against two cell lines, human hepatoblastoma(HepG-2) and human breast carcinoma(MCF-7) by SRB method. The result showed only inonotusin A had moderate cytotoxicity against MCF-7with IC50values of19.6μM, and other four compounds were noncytotoxic to the two cell lines. In vivo anti-tumor activity of petroleum ether and water extracts from fruiting bodies in different stage also carried out by H22tumor-bearing mice transplanting model. The result showed that petroleum ether and water extracts in maturation stage had significant antitumor activities, and the inhibitory value were44.40%(petroleum ether low dosage group,50mg/Kg),55.24%(petroleum ether high dosage group,100mg/Kg),54.01%(water low dosage group,500mg/Kg),52.19%(water high dosage group,1000mg/Kg), and closed to the inhibition of CTX group (61.36%). The water extracts could regulate the immune function of tumor-bearing mice, but petroleum ether extract had effection to immune organ negatively. In darkening fruiting body stage, the inhibitory effects of petroleum ether and water extracts were more lower than the maturation stage, and the value were34.73%(50mg/Kg) and29.99%(500mg/Kg), but the high dosage group had no activity to H22. The inhibition value of methanol extracts was29.77%(500mg/Kg). The extracts had the similar effects to immune organ of tumor-bearing mice, the water extracts could regulate the immune function, but petroleum ether extracts had effection to immune organ negatively. The leucocyte, spleen index and thymus index of water extracts increased higher than CTX group. Moreover, the concentrations of phenolic compounds in the70%ethanol extract of Inonotus hispidus were measured with Folin-Ciocalteu reagent, and total flavonoid concentration was determined by the NaNO2-Al(NO3)3method. The result showed the phenolics were64.02±0.73μg/mg(gallic acid) while total flavanoids were5.74±0.13μg/mg (rutin equivalent) in the70%extract. Antioxidant and antimicrobial activities of Ⅰ. hispidus extracts obtained with70%ethanol and compound hispidin were investigated in this study.·DPPH and·OH free radical-scavenging activity were found to90%inhibition at concentration of200μg/mL. Compound inonotusin B potently scavenged the ABTS radical cation and reached to the level of59.53±9.70μmol.70%ethanol extract and hispidin showed narrow antibacterial activities against Staphyloccocus aureus and Bacillus subtilis.70%ethanol extracts inhibited the growth of the Escherichia coli, and inhibition values was40.35%at concentration of40.96mg/mL.
Fomitiporia ellipsoidea was first collected in Wanmulin Nature Reserve in2008by Bao-Kai Cui and Yu-Cheng Dai, and local people called "sanghuang". In2010, Cui and Dai were performing field work in tropical woodland on Hainan Island, China, studying wood-rotting fungi. The pair uncovered a very large F. ellipsoidea fruit body on a fallen Quercus asymetrica log, which turned out to be the largest fungal fruit body ever documented. Thirteen compounds were isolated from the fruiting body of F. ellipsoidea, and twelve compounds were elucidated, including fomitiporiaester A,(E)-4-(3,4-dihydroxyphenyl) but-3-en-2-one, hispidin, hispolon, inoscavin A, inoscavin C, methylinoscavin D, inoscavin E, phelligridin K, inonoblins B, protocatechuic acid and protocatechualdehyde. Compounds fomitiporiaester A、inoscavin C and inonoblins B were tested for cytotoxic activity against two cell lines, human hepatoblastoma(HepG-2) and human oophoroma (SKOV3) by SRB method. The result showed that inoscavin C and inonoblins B had moderate cytotoxicity against the two cell lines and fomitiporiaester A was noncytotoxic to the two cell lines. In vivo anti-tumor activity of fomitiporiaester A was carried out by H22tumor-bearing mice transplanting model. The result indicated fomitiporiaester A had manifest antitumor activity, and the inhibition ration were 42.94%,49.17%and58.15%at concentration of5mg/Kg,10mg/Kg and20mg/Kg, respectively. Inhibition value nearly to the CTX group (52.56%) at dosage10mg/Kg and better than the CTX group at dosage10mg/Kg. Inhibition effects had dose-effect relationship, and it had significant difference to compare negative control (p<0.01), In addition,compound can increase the immune organ index. Some of isolated hispidin derivates from fruiting bady of F. ellipsoidea potently scavenged the ABTS radical cation in which new compound phelligridin K showed the highest activity at the level of3.56mM Trolox. But fomitiporiaester A had no activity to scavenge the ABTS radical cation.
As a conclusion, the chemical constituents and bioactivities of Ⅰ. hispidus and F. ellipsoidea were studied in this thesis. We isolated some compounds with anti-tumor and antioxidant activitives, including four new compounds.The present results established the basis for discovery of bioactive lead compounds and provided theoretical foundation for further utilization of the two species fungi.
引文
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