南极海洋真菌Penicillium chrysogenum PR4-1-3的活性次级代谢产物研究

目的对一株具有明显细胞毒活性的南极海洋真菌Penicillium chrysogenum PR4-1-3的活性成分进行研究。方法采用溶剂萃取、柱色谱层析及制备HPLC等方法对菌株发酵产物进行活性追踪分离,通过理化性质及波谱学手段进行化学结构鉴定,以SRB法评价化合物的抗肿瘤活性,采用CPE方法对化合物1和2进行抗H1N1甲型流感病毒活性测试。结果从中分离得到5个芳香酚醌类化合物,其结构分别鉴定为secalonic acid D(1)s、ecalonic acid F(2)、chrysophanol(3)、emodin(4)和citreorosein(5)。这几个化合物均具有不同程度的细胞毒活性。化合物1在50μg.mL-1时,对H1N1病毒的抑制率为50%,具有一定的抗病毒活性。结论以上化合物均为首次从南极海洋微生物中分离得到,并初步发现secalonic acid D具有一定的抗H1N1病毒活性。     

红树林耐酸真菌OUCMDZ-4736次生代谢产物研究

目的 探究红树林耐酸真菌OUCMDZ-4736的次生代谢产物及其抗甲型流感病毒H1N1活性。方法 菌株在pH 2.5条件下规模发酵,通过现代色谱学方法（硅胶柱色谱、凝胶柱色谱、半制备以及制备型HPLC）对发酵产物进行分离,运用现代波谱学技术（紫外、质谱、核磁共振等）鉴定分离获得的化合物结构,采用MTT法评价化合物的抗流感病毒H1N1的活性。 结果 从红树林根部泥土样品来源的真菌OUCMDZ-4736中分离鉴定了10个化合物：asterric acid（1）、questinol（2）、parietinic acid（3）、endocrocin（4）、isorhodoptilometrin（5）、sulochrin（6）、monochlorsulochrin（7）、dihydrogeodin（8）、yicathin C（9）、2, 5-dimethyl-6, 8-dihydroxy-chromone（10）。活性评价结果首次报道化合物1、3、4和7显示有弱的抗甲型流感病毒H1N1活性。结论 红树林耐酸真菌OUCMDZ-4736在酸性调节下能代谢产生具有抗甲型流感H1N1病毒活性的化合物。     

深海来源的微生物抗肿瘤活性筛选及次级代谢产物的初步研究

目的 对来自深海的海水、海泥样品进行了微生物分离并通过抗肿瘤活性筛选获得活性菌株,并研究活性菌株c2b的次级代谢产物.方法 从样品中选择性分离得到真菌,并采用海虾生物致死法和人体慢性艇性白血病细胞(K562)为筛选模型对分离得到真菌的发酵产物进行抗肿瘤活性筛选;采用溶剂萃取、硅胶柱色谱及制备HPLC等分离手段对c2b菌株发酵产物的活性部位进行了活性追踪分离,通过理化性质及渡谱学手段进行化学结构鉴定,以SRB法评价了化合物的抗肿瘤活性.结果与结论 从深海来源的样品中共分离获得29株真菌,其中7株具有细胞毒活性;从c2b活性菌株的发酵产物中分离得到6个单体化合物(1～6),其化学结构分别鉴定为N-乙酰色氨(1),chrysogine(2),过氧化麦角甾醇(3),5,8-epidioxy-24-methylcholesta-6,22-dien-3β-ol(4),cerevisterol(5)和(4E,8E)-N-[(2'R,3'E)-2'-hydroxy-3'-hexadecenoyl]-1-O-β-D-glycopyranosyl-9-methyl-4,8-sphingadiene(6),其中化合物3,4对小鼠乳腺癌细胞(tsFT210)具有中等强度的细胞毒活性.     

海洋真菌烟曲霉H1-04生产的生物碱类化合物及其抗肿瘤活性

目的研究海洋真菌烟曲霉H1-04（Aspergillus fumigatus H1-04）生产的抗肿瘤活性产物。方法以生物活性为导向．利用液液萃取、硅胶柱色谱、制备HPLC等技术分离纯化活性化合物；根据理化常数及波谱数据鉴定化学结构：采用SRB法和MTT法测试评价抗肿瘤活性。结果与结论从烟曲霉H1—04发酵物中分得7个化合物，分别鉴定为fumiquinazoline J（1）、fumiquinazoline F（2）、fumiquinazoline G（3）、fumiquinazoline C（4）、fumiquinazoline A（S）、tryptoquivaline J（6）和pseumtin A（7）。化合物1～7对小鼠乳腺癌tsFT210细胞均呈一定的细胞增殖抑制活性，1、4和7对人肝癌BEL-7402、人肺癌A549、人白血病HL60和小鼠白血病P388细胞也有不同程度的抑制活性。化合物1为首次从自然界分离获得，其抗肿瘤活性也属首次报道，化合物6为首次从海洋来源微生物产物中分离得到。     

海洋真菌烟曲霉H1-04发酵液中的硫代二酮哌嗪类产物及其抗肿瘤活性

目的 研究海洋来源真菌烟曲霉H1-04(Aspergillus fumigatus H1-04)发酵液中的抗肿瘤活性产物.方法 采用液液萃取、硅胶柱色谱、制备HPLC等技术,从烟曲霉H1-04的发酵产物中分离纯化活性产物.活性产物的结构经IR、MS、1H-NMR确证.采用SRB法和MTT法测试评价抗肿瘤活性.结果与结论 从烟曲霉H1-04发酵物中分离鉴定了4个硫代二酮哌嗪类化合物,分别是胶霉毒素(gliotoxin,1)、bisdethiobis(methylthio)gliotoxin(2)、bis-N-norgliovictin(3)、didehydrobisdethiobis(methylthio)gliotoxin(4).化合物1在浓度为0.1μmol·L-1时几乎能完全抑制小鼠白血病P388细胞和人肺癌A549细胞的增殖,抑制率分别为100%和99.1%;对小鼠乳腺癌tsFT210细胞也呈现很强的细胞凋亡诱导、细胞周期抑制、坏死性细胞毒等活性.化合物2～4对tsFT210细胞显示出不同程度的抗肿瘤活性.化合物2～4为首次从烟曲霉发酵物中分离得到,并首次报道2～4的抗肿瘤活性.     

海洋来源微生物的分离培养与抗肿瘤活性筛选

目的从海洋环境样品中分离纯化微生物,经发酵培养与活性筛选,获取抗肿瘤活性菌株以供筛选药源活性产物,获无活性菌株以供核糖体工程转化研究。方法通过单菌落挑选与划线培养分离纯化微生物菌株,经摇床发酵和提取操作制备活性测试样品。采用MTT法结合显微镜下细胞形态学检测的方法,测试样品的抗肿瘤活性。结果从渤海湾驴驹河潮间带海泥样品中分离得到了微生物127株,其中真菌80株、放线菌47株。在127株中100mg·L-1样品浓度下对K562细胞的抑制率大于40%的活性菌株为8株,占菌株总数的6.3%(其中放线菌4株,占放线菌数的8.5%,真菌4株,占真菌数的5%),抑制率在20%～40%的放线菌6株,占放线菌数的12.7%。结论从渤海湾海泥样品中分离得到真菌80株、放线菌47株,从中获得抗肿瘤活性真菌4株、放线菌10株,从放线菌中获得抗肿瘤活性菌株的频率远远高于真菌。活性菌株为寻找药源活性产物提供了菌株,无活性菌株则为核糖体工程拓展药源菌株来源研究提供了资源。     

1株黄河三角洲来源杂色曲霉A.versicolor BHT-72的次级代谢产物研究

目的 研究1株黄河三角洲盐碱地曲霉属真菌A. versicolor BHT-72的次级代谢产物及其生物活性。方法 采用硅胶柱层析、Sephadex LH-20葡聚糖凝胶柱层析、ODS柱层析、高效液相（HPLC）等色谱方法对该菌株的次级代谢产物分离纯化,并通过核磁共振（NMR）、质谱（MS）等方法,结合相关文献对比鉴定化合物结构;分别采用CCK8法和改良的Ellman法对化合物进行抗肿瘤和抗乙酰胆碱酯酶抑制活性的测试。结果 从A. versicolor BHT-72的大米发酵产物中共分离得到10个单体化合物,分别为diorcinol(1)、12-O-acetyl-sydowinin A(2)、sydowinin A(3)、13-O-acetylsydowinin B(4)、sydoxanthone(5)、3-formylindole(6)、尿苷（7）、zarzissine(8)、6-氨基嘌呤核苷（9）、6-氨基嘌呤脱氧核苷（10）。细胞毒活性测定结果显示,化合物1在质量浓度为30μg/mL时,对鼻咽癌细胞CNE-2的抑制率为65.0%,对非小细胞肺癌H1299和H520的抑制率分别为81...     

抗甲型流感H1N1病毒红树林内生放线菌的筛选及菌株HA12210的鉴定

目的对红树林内生放线菌进行抗H1N1病毒活性检测,并对具有较高活性的菌株HA12210进行鉴定。方法采用嗜杀酵母系统和CPE+MTT联合法模型对菌株发酵液进行抗H1N1病毒活性的初筛和复筛;对菌株HA12210进行培养和形态特征、生理生化特征的鉴定,测定其16SrDNA序列并进行系统发育分析。结果菌株HA12210发酵液稀释20倍后对H1N1病毒的抑制率达到73.2%。HA12210与Micromonospora marina JSM1-1T形态和生理生化特征接近,并与其16SrDNA序列相似性最高(99.70%),且在发育树上聚为同一分支。结论鉴定活性菌株HA12210为Micromonospora marina,本文首次报道了该菌株的抗H1N1病毒活性。     

海洋放线菌分离及菌株Streptomyces sp. AH17-3的次级代谢产物研究

目的对海洋放线菌进行分离及抗肿瘤活性筛选,并对一株具有抗肿瘤活性的海洋放线菌AH17-3的次级代谢产物进行研究。方法采用溶剂萃取、柱色谱层析及制备HPLC等方法对菌株AH17-3的发酵产物进行化学分离,通过理化性质及波谱学方法并参阅文献进行化合物结构鉴定,以SRB法评价化合物的抗肿瘤活性。结果从海洋样品中分离放线菌174株,从菌株AH17-3中分离得到了4个聚酮类化合物,经鉴定其结构分别为germicidin A(1)、germicidin B(2)、daidzein(3)、genistein(4)。其中化合物1具有弱的细胞毒活性,其IC50为3.5×10-7 M。结论海洋放线菌是重要的药用微生物资源,化合物1,2均为首次从海洋放线菌中分离得到。     

海绵来源真菌黄灰青霉Sp-19中的抗肿瘤活性成分研究

目的对一株来源于羽毛山海绵Mycale plumose的真菌黄灰青霉Penicillium auratiogriseumSp-19发酵产物中的抗肿瘤活性成分进行分离和鉴定。方法采用溶剂萃取、硅胶柱色谱及制备HPLC等分离手段对该菌株发酵产物的活性部位进行了活性追踪分离,通过理化性质及波谱学手段进行化学结构鉴定,以SRB法评价了化合物的抗肿瘤活性。结果与讨论从发酵产物中分离得到5个生物碱类化合物,其结构分别鉴定为fructigenines A(1),去乙酰化fructigenines A(2),fructigeninesB(3),1,4-benzodiazepine-2,5-diones(4)和cyclopenin(5);化合物4和5在3μg.mL-1时对小鼠乳腺癌tsFT210细胞具有强的细胞毒活性。     

Antimicrobial activity and cytotoxicity of bis(indole) alkaloids from the sponge Spongosorites sp

Bis(indole) alkaloids, of the topsentin class (1-4) and hamacanthin class (5-9), isolated from the marine sponge Spongosorites sp. were investigated using several biological assays. In the evaluation of antimicrobial activity against various strains of bacteria and fungi, compounds of the hamacanthin class exhibited more potent antibacterial activity than those of the topsentin class. Deoxytopsentin (1) and hamacanthin A (5) also exhibited significant antibacterial activity against methicillin-resistant Staphylococcus aureus, with MIC values of less than 12.5 microg/ml. In the antifungal activity test, hamacanthins, especially hamacanthin A (5), showed potent inhibitory activity against medically important pathogenic fungi. In contrast, all of the topsentins (1-4) were inactive against fungal growth. These compounds (1-9) also exhibited moderate cytotoxicity against cancer cell lines at concentrations between 1.1 and >20 microg/ml.     

大狼毒内生真菌的分离、鉴定与抑菌活性研究

目的 分离和鉴定大狼毒根中的内生真菌,并检测其抑菌活性.方法 用组织块法分离,点植法对分离菌株进行分类鉴定;选择3种人类病原细菌作为指示菌进行体外抑菌试验.结果 分离获得内生真菌41株,鉴定为1纲、4目、6科、15属;其中9株有稳定的抑菌活性,尤其对大肠杆菌有显著抑制作用.结论 大狼毒根中存在丰富的内生真菌,其代谢产物具有一定的抑菌作用,可作为筛选抑菌活性物质的新资源.     

海洋来源青霉菌Penicillium polonicum H92次级代谢产物的研究

目的 对源自福建漳江口红树林底部海水沉积物的青霉菌Penicillium polonicum H92的次级代谢产物进行化学成分和生物活性研究.方法 综合利用有机溶剂萃取、硅胶柱色谱、反相硅胶(ODS)柱色谱、凝胶(Sephadex LH-20)柱色谱、薄层色谱、半制备高效液相色谱等多种手段对青霉菌Penicillium...     

中国东海及东太平洋海山区来源抗菌活性海洋微生物的筛选研究

摘要：目的 研究中国东海及东太平洋区域来源微生物的抗菌活性,意在筛选具有良好抗菌活性的海洋微生物,并初步研究活性菌株代谢产物的结构,为探求新型抗生素提供基础。方法 以金黄色葡萄球菌、大肠杆菌和白色念珠菌为指示菌株,采用牛津杯法进行抗菌活性微生物初筛和复筛。通过HPLC-DAD结合TLC对活性突出的4株真菌及1株细菌进行活性提取物指纹图谱分析。对广谱抗菌活性菌株WBX-38进行菌种鉴定及活性代谢产物分离纯化,利用核磁共振和质谱等手段对活性化合物进行结构鉴定。采用微量稀释法测定活性化合物的最小抑菌浓度（MIC）。结果与结论 从中国东海及东太平洋来源样品中共筛选90株海洋来源菌株,获得抗菌活性菌株16株,4株为海洋细菌,12株为海洋真菌。通过HPLC-DAD结合TLC分析,发现5株活性菌株均具有独特的色谱行为。海洋真菌WBX-38通过菌种鉴定,确认为海洋来源曲霉（Aspergillus sp.）。从其发酵液提取物中分离获得1个活性化合物5-hydroxymethylfuran-3-carboxylic acid,该化合物对3种指示菌均具有一定生长抑制活性。     

2株海洋真菌共培养产青霉酸及其抗菌活性研究

目的研究2株海洋真菌Penicillium janthinellum HK1-6与Trichoderma reesei HK1-12共培养体系中的次级代谢产物变化及其抗菌活性。方法采用代谢组学方法分析共培养中的差异化合物,采用高效液相法对共培养中的青霉酸进行定性及定量分析,采用带毒培养基法和滤纸片法评价青霉酸的抗真菌及抗人类致病细菌活性。结果有97个差异化合物仅在真菌HK1-6与HK1-12共培养体系产生;真菌HK1-6在共培养刺激下分泌青霉酸的产量增加,较HK1-6纯培养时提高约1.2倍;青霉酸对真菌HK1-12和5株人类致病细菌均具有较强抗菌作用。结论共培养策略可以激活真菌某些沉默的代谢途径或提高某些代谢产物的产量,是挖掘海洋真菌次级代谢产物的有效途径。     

Phenolic compounds from Baseonema acuminatum leaves: isolation and antimicrobial activity

Three new phenolic compounds, 1-galloyl-beta-D-glucopyranosyl-(1-->4)-beta-D-galactopyranoside (1), 2-methoxy-5-(1 '2 3'-trihydroxypropyl)-phenyl- 1-0-(6"-galloyl)-beta-D-glucopyranoside (2),and 2-methoxy-5-hydroxymethyl-phenyl-1-O-(6"-galloyl)-beta-D-glucopyranoside (3), together with the known compounds benzyl 6'-O-galloyl-beta-D-glucopyranoside (4), 1,6-di-O-galloyl-beta-D-glucopyranose (5), myrciaphenone B (6), kaempferol 3-0-(6"-galloyl)-beta-D-glucopyranoside (7), quercetin 3-0-(6"-galloyl)-beta-D-glucopyranoside (8), vomifoliol 9-O-beta-D-apiofuranosyl-(1-->6)-beta-D-glucopyranoside, 2,3-dihydrobenzofuran-2-(4'-hydroxy-3'-methoxyphenyl)-3-alpha-L-rhamnopyranosyloxymethyl-7-methoxy-5-propanol, and benzyl-O-alpha-L-rhamnopyranosyl-(1-->6)-Beta-D-glucopyranoside were isolated from the leaves of Baseonema acuminatum P. Choux (Asclepiadaceae). Their structures were determined by 1D- and 2D-NMR spectroscopy and by ESI-MS analysis. The antimicrobial activity of all compounds was evaluated in vitro against bacteria (Staphylococcus aureus two strains, Bacillus cereus, Bacillus subtilis, Escherichia coli, Salmonella thyphimurium) and three strains of Candida albicans. The new compounds 2 and 3, together with the known compound 4, showed antifungal activity against two clinically isolated Candida albicans strains and against C. albicans ATCC 2091; MIC values were in the range of 25-100 microg/mL. Compound 5 was active against the two clinically isolated strains of C. albicans with MICs of 12.5 microg/mL and 25 microg/mL. Compounds 1, 6, 7, and 8 inhibited only one strain of C albicans at the maximum concentration used. None of the phenolic compounds tested was active against the bacteria studied.     

Brosimacutins J-M, four new flavonoids from Brosimum acutifolium and their cytotoxic activity

Four new flavonoids, brosimacutins J-M (1 - 4), were isolated from the bark of Brosimum acutifolium Huber together with a known flavan, brosimine A (5). The structures of compounds 1-4 were elucidated by spectroscopic means. 27 constituents of this plant including compounds 1-5 were evaluated for their cytotoxic activity against murine leukemia P388 cells. Although no compounds tested had any reversal effect on vincristine resistance, brocimacutins J-M (1-4) were cytotoxic to vincristine-resistant P388 cells (IC50 4.4 - 19 microg/mL).     

Synthesis and antitumor and antiviral activities of a series of 1-beta-D-ribofuranosyl-5-halocytosine (5-halocytidine) cyclic 3',5'-monophosphates

A series of 1-beta-ribofuranosyl-5-halocytosine cyclic 3',5'-monophosphates (1-4) has been prepared. Direct halogenation of cytidine 3',5'-monophosphate (cCMP) yielded the Cl, Br, and I compounds while 5-F-cCMP (1) was obtained on cyclization of the 5'-monophosphate. On in vitro testing of 1-4 against L1210 and P388 leukemias, only 1 showed significant low-level activity (ID50 = 3.1 X 10(-4) mmol/L). Derivatives 2-4 were inactive at 10(-1) mmol/L and also proved to have low viral ratings against a series of RNA and DNA virus strains in vitro. By contrast the 5-F-cCMP showed moderate activity against VV, HSV-1, and HSV-2 strains (VR = 0.6-0.9). Both 5-fluorocytidine and 5-fluorocytidine 5'-monophosphate had marked antiviral activity (VR = 1.0-2.1) with the above viruses as well as with parainfluenza virus type 3. The nucleoside and nucleotide also were more active than 5-F-cCMP against L1210 and P388 cells. However, comparison of the cytotoxicities and antiviral ED50 values of 5-F-cCMP, 5-fluorocytidine 5'-monophosphate, and 5-fluorocytidine suggests a potential therapeutic advantage for 5-F-cCMP. Possible rationales for these activities are discussed in terms of 5-F-cCMP and the corresponding 5'-monophosphate as potential prodrugs and as sources, following enzymatic deamination, of cytotoxic 5-fluorouridine or its 5'-monophosphate.     

海洋真菌zp6次级代谢产物及其抗真菌活性研究

目的对海洋真菌zp6发酵液的乙酸乙酯萃取部分及其抑菌活性进行研究。方法采用硅胶柱色谱及高效液相色谱进行分离纯化,根据理化性质及各种光谱技术进行结构鉴定,采用滤纸片法对分离到的化合物进行抑菌活性研究。结果从该菌中分离纯化得到8个化合物:(22E)-3β,5α,9α-三羟基-麦角甾-7,22-二烯-6-酮(1)、5α,8α-环二氧-24-甲基麦角甾-6,22-二烯-3β-醇(2)、赤藓糖醇(3)、腺嘌呤核苷(4)、(24R)-麦角甾-7,22-二烯-3β,5α,6β-三醇(5)、阿拉伯醇(6)、(7)、4-乙酰氧基苯甲酸(8)。结论化合物1和5具有抗真菌活性。     

Metabolites of febrifugine and its synthetic analogue by mouse liver S9 and their antimalarial activity against Plasmodium malaria parasite

Quinazolinone type alkaloids, febrifugine (1) and isofebrifugine (2), isolated from Dichroa febrifuga roots, show powerful antimalarial activity against Plasmodium falciparum. Unfortunately, their emetic effect and other undesirable side effects have precluded their clinical use for malaria. Because of their antimalarial potency, analogues were searched for, with the goal of preserving the strong antimalarial activity, while dramatically reducing side effects. We expected that compounds useful in drug development would exist in metabolites derived from 1 and Df-1 (3), the condensation product of 1 with acetone, by mouse liver S9. Feb-A and -B (4 and 5) were isolated as the major metabolites of 1. In addition to 4 and 5, feb-C and -D (6 and 7) were also purified from the metabolic mixture of 3. Compounds 4 and 5 were compounds oxidized at C-6 and C-2 of the quinazolinone ring of 1, respectively. Compounds 6 and 7, derived from 3, also bear febrifugine type structures in which the 4' '- and 6' '-positions of the piperidine ring of 1 were oxidized. In vitro antimalarial and cytotoxic tests using synthetically obtained racemic 4-6 and enantiomerically pure 7 demonstrated that 4 and 6 had antimalarial activity against P. falciparum, of similar potency to that of 1, with high selectivity. The antimalarial activity of 5 and 7, however, was dramatically decreased in the test. The in vitro antimalarial activity of analogues 22 and 43, which are stereoisomers of 4 and 6, was also evaluated, showing that 22 is active. The results suggest that basicity of both the 1- and the 1' '-nitrogen atoms of 1 is crucial in conferring powerful antimalarial activity. Racemic 4 and 6 exhibited powerful in vivo antimalarial activity against mouse malaria P. berghei, and especially, no serious side effects were observed with 4. Thus, the metabolite 4 appears to be a promising lead compound for the development of new types of antimalarial drugs.