药用植物细圆藤和三株放线菌抗生素抗性突变株的代谢产物及其抗肿瘤活性
|
摘要
从天然产物中寻找抗肿瘤活性先导化合物对于开发新型抗癌药物具有积极的意义。本论文对药用植物细圆藤与三株放线菌抗生素抗性突变株的代谢产物及其抗肿瘤活性进行了研究。研究内容主要包括:细圆藤化学成分的提取分离、从无活性放线菌筛选获取活性突变株与三株放线菌活性突变株代谢产物的分离纯化、单体化合物的结构鉴定以及其抗肿瘤活性初步测试等。
论文前半部分是关于细圆藤的化学成分及其抗肿瘤活性研究。细圆藤在《中华药海》中记录有治疗无名肿毒、疥疮肿毒、毒蛇咬伤、小儿惊风、破伤风等功效。鉴于前期研究中发现细圆藤枝叶提取物具有体外抗肿瘤活性而活性成分未见报道,本论文前半部分对细圆藤的化学成分进行了分离纯化、结构鉴定以及抗肿瘤活性的初步测试评价。结果从细圆藤枝叶中分离鉴定了五个化合物hopenone-B(1)、hopenol-B(2)、22-Hydroxyhopan-3-one(3)、erythrodiol 3-palmitate(4)和5β,24-cyclofriedelan-3-one(5)。其中,化合物4具有较强的抗肿瘤活性,在100μg/mL浓度下对K562细胞的抑制率为46.7%。本论文上述化合物1～5均为首次从该属中分离得到,其中化合物4为该植物抗肿瘤活性成分的首例报道,其抗肿瘤活性也属首次筛选发现。
论文后半部分是有关从无活性放线菌野生菌株筛选获取活性突变株以及有关三株抗肿瘤活性突变株代谢产物及其抗肿瘤活性的研究内容。论文首先以一株没有抗肿瘤活性的海洋放线菌M15为出发菌株,采用抗生素抗性筛选技术即核糖体工程技术,开展了链霉素、新霉素、庆大霉素、利福平等抗生素抗性菌株筛选获取工作,从而获得了对不同抗生素产生抗性的突变株63株,再用突变株发酵产物采用MTT法筛选了抗肿瘤活性,结果得到有明显活性的突变株6株。继而,选择其中3株活性突变株,采用活性跟踪和薄层对照的方法,对与原始菌比较有差异的产物进行了跟踪分离,从中分离得到9个化合物,并分别鉴定为尿嘧啶(6)、染料木素(7)、对-甲酰胺苯甲酸(8)、2-吡咯酸(9)、邻二羟基苯甲酸(10)、过氧化麦角甾醇(11)、N-(2-羟基苯基)-乙酰胺(12)、大豆黄素(13)和1,9-二甲酯-吩嗪(14)。活性测试结果表明,化合物6、7、11、13和14对K562细胞有抑制作用,在100μg/mL浓度下的抑制率分别为41.4%、51.6%、28.1%、21.4%和25.2%。其中14为新天然产物,其抗肿瘤活性也属首次筛选发现。本论文上述研究结果表明,核糖体工程技术是一种有效的开拓药源微生物菌株资源的新方法。
This dissertation describes the secondary metabolites and their anti-tumor activity from Pericampylus galucus and three actinomycetic antibiotic-resistent atrains.Studies include the isolation,identification and anti-tumor activities of the constituents from Pericampylus galucus,as well as the induction of a non-active actinomycetic strain,fermentation,screening of antibiotic-resistent Strains by MTT method using K562 cells and the isolation, identification and anti-tumor activities of the constituents from three actinomycetic antibiotic-resistent Strains.
The front segment of dissertation is about isolation and identification of the constituents from Pericampylus galucus and their anti-tumor activities.The bronches and leaves of Pericampylus galucus has been used as a folk medicine in Yunnan province of China to cure tumor,itch,tetanus,eclampsia et al.The alcohol extract of its bronches and leaves with anticancer activity screened by MTT method showed anti-tumor activities.In order to elucidate the anti-tumor constituents,the chemical and biological investigation had been carried out.The results are fellow as:five triterpenoids 1-5 were isolated from Pericampylus glaucus and they were identified as Hopenone-B(1),hopenol-B(2),22-Hydroxyhopan-3-one (3),erythrodiol 3-palmitate(4)and 5β,24-cyclofriedelan-3-one(5);Among them,4 inhibited the proliferation of K562 cells with the inhibition rate of 47%at 100μg/mL; Compounds 1-5 were isolated from the genus Pericampylus for the first time,4 provides the first example of anti-tumor constituents of Pericampylus glaucus and its anti-tumor activity was also assayed for the first time.
The rear segment of dissertation is about the method of a non-active Actinomycetic strain chosen as a original strain and induced the antibiotic resistant,isolation and identification of the constituents from three actinomycetic antibiotic-resistent Strains,preliminary evaluation for anti-tumor activities of pure compounds.A strain M15 shows no anti-tumor activities and was chosen as a original strain and then was induced the antibiotic(streptomycin,neomycin, gentamicin,Rifampicin)resistant i.e.ribosome engineering technology.63 strains of mutation were obtained,and their fermentation with anti-tumor activity screened by MTT method were examined using K562 cells.6 strains of 63 shows anti-tumor activities.3 of 6 strains having anti-tumor activities was to elucidate the anti-tumor constituents with the method of Guiding by bioassay of anti-tumor and comparing with original strain.The results are fellow as:9 compounds 6-14 were isolated and identified as uracil(6),Genistein(7),4 -(methyamino)-benzoic acid(8),1H-pyrrole-2-canboxylic acid(9),2,3-dihydrobenzoic acid (10),ergosterol peroxide(11),N-(2-hydroxyphenyl)-acetamide(12),Daidzein(13), 1,9-Dicarbomethoxy-phenazine(14).6,7,11,13 and 14 inhibited the proliferation of K562 cells with the inhibition rate of 41.4%,51.6%,28.1%,21.4%and 25.2 at 100μg/mL; Compounds 14 were isolated from the nature for the first time and its anti-tumor activity was also assayed for the first time.Above studies indicated that it is feasible to exploit new microbial resources for medicinal usage by applying the ribosome-engineering technology to non-active actinomycete in nature.
论文前半部分是关于细圆藤的化学成分及其抗肿瘤活性研究。细圆藤在《中华药海》中记录有治疗无名肿毒、疥疮肿毒、毒蛇咬伤、小儿惊风、破伤风等功效。鉴于前期研究中发现细圆藤枝叶提取物具有体外抗肿瘤活性而活性成分未见报道,本论文前半部分对细圆藤的化学成分进行了分离纯化、结构鉴定以及抗肿瘤活性的初步测试评价。结果从细圆藤枝叶中分离鉴定了五个化合物hopenone-B(1)、hopenol-B(2)、22-Hydroxyhopan-3-one(3)、erythrodiol 3-palmitate(4)和5β,24-cyclofriedelan-3-one(5)。其中,化合物4具有较强的抗肿瘤活性,在100μg/mL浓度下对K562细胞的抑制率为46.7%。本论文上述化合物1～5均为首次从该属中分离得到,其中化合物4为该植物抗肿瘤活性成分的首例报道,其抗肿瘤活性也属首次筛选发现。
论文后半部分是有关从无活性放线菌野生菌株筛选获取活性突变株以及有关三株抗肿瘤活性突变株代谢产物及其抗肿瘤活性的研究内容。论文首先以一株没有抗肿瘤活性的海洋放线菌M15为出发菌株,采用抗生素抗性筛选技术即核糖体工程技术,开展了链霉素、新霉素、庆大霉素、利福平等抗生素抗性菌株筛选获取工作,从而获得了对不同抗生素产生抗性的突变株63株,再用突变株发酵产物采用MTT法筛选了抗肿瘤活性,结果得到有明显活性的突变株6株。继而,选择其中3株活性突变株,采用活性跟踪和薄层对照的方法,对与原始菌比较有差异的产物进行了跟踪分离,从中分离得到9个化合物,并分别鉴定为尿嘧啶(6)、染料木素(7)、对-甲酰胺苯甲酸(8)、2-吡咯酸(9)、邻二羟基苯甲酸(10)、过氧化麦角甾醇(11)、N-(2-羟基苯基)-乙酰胺(12)、大豆黄素(13)和1,9-二甲酯-吩嗪(14)。活性测试结果表明,化合物6、7、11、13和14对K562细胞有抑制作用,在100μg/mL浓度下的抑制率分别为41.4%、51.6%、28.1%、21.4%和25.2%。其中14为新天然产物,其抗肿瘤活性也属首次筛选发现。本论文上述研究结果表明,核糖体工程技术是一种有效的开拓药源微生物菌株资源的新方法。
This dissertation describes the secondary metabolites and their anti-tumor activity from Pericampylus galucus and three actinomycetic antibiotic-resistent atrains.Studies include the isolation,identification and anti-tumor activities of the constituents from Pericampylus galucus,as well as the induction of a non-active actinomycetic strain,fermentation,screening of antibiotic-resistent Strains by MTT method using K562 cells and the isolation, identification and anti-tumor activities of the constituents from three actinomycetic antibiotic-resistent Strains.
The front segment of dissertation is about isolation and identification of the constituents from Pericampylus galucus and their anti-tumor activities.The bronches and leaves of Pericampylus galucus has been used as a folk medicine in Yunnan province of China to cure tumor,itch,tetanus,eclampsia et al.The alcohol extract of its bronches and leaves with anticancer activity screened by MTT method showed anti-tumor activities.In order to elucidate the anti-tumor constituents,the chemical and biological investigation had been carried out.The results are fellow as:five triterpenoids 1-5 were isolated from Pericampylus glaucus and they were identified as Hopenone-B(1),hopenol-B(2),22-Hydroxyhopan-3-one (3),erythrodiol 3-palmitate(4)and 5β,24-cyclofriedelan-3-one(5);Among them,4 inhibited the proliferation of K562 cells with the inhibition rate of 47%at 100μg/mL; Compounds 1-5 were isolated from the genus Pericampylus for the first time,4 provides the first example of anti-tumor constituents of Pericampylus glaucus and its anti-tumor activity was also assayed for the first time.
The rear segment of dissertation is about the method of a non-active Actinomycetic strain chosen as a original strain and induced the antibiotic resistant,isolation and identification of the constituents from three actinomycetic antibiotic-resistent Strains,preliminary evaluation for anti-tumor activities of pure compounds.A strain M15 shows no anti-tumor activities and was chosen as a original strain and then was induced the antibiotic(streptomycin,neomycin, gentamicin,Rifampicin)resistant i.e.ribosome engineering technology.63 strains of mutation were obtained,and their fermentation with anti-tumor activity screened by MTT method were examined using K562 cells.6 strains of 63 shows anti-tumor activities.3 of 6 strains having anti-tumor activities was to elucidate the anti-tumor constituents with the method of Guiding by bioassay of anti-tumor and comparing with original strain.The results are fellow as:9 compounds 6-14 were isolated and identified as uracil(6),Genistein(7),4 -(methyamino)-benzoic acid(8),1H-pyrrole-2-canboxylic acid(9),2,3-dihydrobenzoic acid (10),ergosterol peroxide(11),N-(2-hydroxyphenyl)-acetamide(12),Daidzein(13), 1,9-Dicarbomethoxy-phenazine(14).6,7,11,13 and 14 inhibited the proliferation of K562 cells with the inhibition rate of 41.4%,51.6%,28.1%,21.4%and 25.2 at 100μg/mL; Compounds 14 were isolated from the nature for the first time and its anti-tumor activity was also assayed for the first time.Above studies indicated that it is feasible to exploit new microbial resources for medicinal usage by applying the ribosome-engineering technology to non-active actinomycete in nature.
引文
[1]董志伟,乔友林,李连弟.中国癌症控制策略研究报告[J].中国肿瘤,2002,11(5):250-260
[2]潘启超,胥彬,肿瘤药理学与化学治疗学[M].第二版,郑州:河南医科大学出版社,3-10
[3]Proksch P.,Edrada R.A.,Eber.1..Drugs from the seas-Current status and microbiological implications[J].Appl Microbiol Biotechnol,2002,59(223):125-134
[4]刘景生.细胞信息与调控.现代生物医学丛书.北京:北京医科大学、北京协和医科大学联合出版社.1998:318-333
[5]Keer JF,Wyllie AH,Currie AR,Apoptosis,et al,a basic biological phenomenon with wide-ranging implications in tissue kinetics,Br J Cancer.1972,26:239-251
[6]王海燕,蔡兵,崔承彬,等.蔓荆子活性成分vitexicarpin诱导K562细胞凋亡的机制[J].药学学报,2005,40(1):27-31.
[7]Nikaido H.,Prevention of drug acessto bacterial tagets:permeability barriers and active effiux[J].Science,1994,264:382
[8]Spratt B G.,Resistance to antibiotics mediated by target alerration[J].Science,1994,264:388
[9]沈同,王镜岩.生物化学(下册)北京:高等教育出版社2000,398
[10]Finken,M.,P.Kirschner,A.Meier,A.Wrede,and E.C.Bo"ttger.Molecular basis of streptomycin resistance in Mycobacterium tuberculosis:alterations of the ribosomal protein S12 gene and point mutations within a functional 16S ribosomal RNA pseudoknot.Mol.Microbiol.1993.9:1239-1246.
[11]Cundlife E.Recognition sites for antibiotics with in rRNA.In:Hill W E,Dahlberg A,Carret RA,et al.The ribosome:structure,function and evolution.[M].Washington DC:American Society for Microbiology,1990:479
[12]Hima J,Hesketh A,Okamato S,et al.Induction of actinorhodin production by rpsl(encoding ribosomal protein S12)mutation that confer streptomycin resistance in Streptomyces lividans and Streptomyces coelicolor A3(2)[J].Bacteriol,1996,178(24):7276-7284
[13]Hu.H.F,Ochi K.Novel approach for improving the productivity of antibiotic-producing strains by inducing combined resistant mutations.Appl Environ Microbiol,2001,67(4):1885-1892
[14]Hosoya Y,Okamoto S,Muramatsu H et al.Acquisition of certain streptomycin-resistant(str)mutations enhances antibiotic production in bacteria.Antimicrob.Agents.Chemotherapy,1998,42(8):2941-2047
[15]涂国全等.通过获得链霉素抗性基因(str)突变株筛选梅岭霉素高产菌株[J].中国抗生素杂志,2002,27(6):321-325
[16]陶纯长等.链霉素抗性突变理性筛选avermetin高产菌株.中国抗生素杂志[J].2002,27(9):521-523
[17]杨东靖,陈冠群等.纳他霉素高产菌株的链霉素抗性选育及其发酵工艺的优化[J].药物生物技术.2003,10(2):84-87
[18]Yoshiko O H,Susumu O,and Kozo Ochi,Development of Antibiotic-Overproducing Strains by Site-Directed Mutagenesis of the rpsL Gene in Streptomyces lividans,Appl and Environment Microbiology,July 2003,4256-4259
[19]Hopwood D A,Chater K F.Fresh approaches to antibiotic production[M].Philos Trans R Soc Lond B Biol Sci,1980,290(1040):313
[20]顾觉奋,获取抗生素的新途径—沉默基因的激活,国外医药抗生素分册[J].2002,24(5):193-197
[21]Hopwood D A.Towards on understanding of gene switching in Streptomyces,the basis of sporulation and antibiotics production[M]Proc R Soc Lond Biol,1988,235:121
[22]崔承彬,顾谦群等.开拓微生物药用资源暂新领域的新技术方法学研究.第八届中国海洋药物学术研讨会.青岛.2002.45-47
[23]王海燕,蔡兵,崔承彬,等.蔓荆子活性成分vitexicarpin诱导K562细胞凋亡的机制[J].药学学报,2005,40(1):27-31.
[24]连文琰.中国防己科药用植物的初步调查[J].植物分类学报,1975,13(1):32-53.
[25]冉先德.中华药海[M].第一版,哈尔滨:哈尔滨出版社,1993:1925.
[26]梁培瑜,周琦,周法兴.细圆藤的化学成分研究[J].中国中药杂志,1998,23(1):39-40.
[27]李长伟.南酸枣抗肿瘤活性成分的研究[M].沈阳药科大学硕士学位论文,2005:13-17.
[28]CHATURVEDULA V S P,SCHILLING J K,MILLER J S,et al.Two new triterpene esters from the twigs of Brachylaena ramifiora from the Madagascar rainforest[J].J Nat Prod,2002,65(8):1222-1224.
[29]MATUSUNAGA S,MORIITA R.Hopenol-B,A triterpene alcohol from Euphorbia supina[J].Phytochemistry,1983,22(2):605-606.
[30]MCLEAN S,PERPICN-DUMONT M,REYNOLDS W F,et al.Unambiguous structural and nuclear magnetic resonance spectral characterization of two triterpenoids of Maprounea guiqnensis by two-dimensional nuclear magnetic resonance spectroscopy[J].Can J Chem,1987,65(11): 2519-2525.
[3l]TSTUDA Y,HATORI M.Total synthesis of hydroxyhopanone[J].Chem Pharm Bull,1967,15(7):1073-1074.
[32]ROWAN D D,RUSSELL G B.3β-merthoxyhop-22(29)-ene from Chionochloa cheesemanii[J].Phytochemistry,1992,31(2):702-703.
[33]WILKINS A L,RONALDSON K J,JAGER P M,et al.A ~(13)C NMR Study of some oxygenated hopane triterpenes[J].Aust J Chem,1987,40(10):1713-1721.
[34]NAKANO T,HASEGAWA M.Isolation of erythrodiol 3-palmitate from Bumelia obtusifolia[J].Planta Med,1975,27(1):89-91.
[35]WANDJI J,TILLEQUIN F,MULHOLLAND D A,et al.Fatty acid esters of triterpenoids and steroid glycosides from Gambeya africana[J].Planta Med,2002,68(9):822-826.
[36]CONNOLLY J D,FREER A A.Structure of 5β,24-cyclofriedelan-3-one,a novel hexacyclic triterpenoid in space group P1[J].Acta Cryst,1986,C42:1352-1354.
[37]朱峰,林永成,周世宁等,南海红树林内源真菌2534号代谢产物的研究[J].中山大学学报(自然科学版)2003,42(1):52-54
[38]江红,程元荣,郑卫,海洋碳样小单孢菌产生的大豆黄素和染料木素[J].中国海洋药物杂志2007,26(1):8-12
[39]张海龙,田黎,傅红伟等,海洋真菌Alternalia sp.发酵液中化学成分的研究[J].中国中药杂志2005,30(5):351-353
[40]Romer A.~1H NMR spectra of substituted phenazines[J].Org Mag Res,1982,19(2):66-68.
[41]Breitmaier E,Hollstein U.Carbon-13 Nuclear magnetic resonance chemical shifts of substituted phenazines[J].J Org Chem.1976,41(12):2104-2108.
[1]P.Proksch,R.A.Edrada,r.Ebel.Drugs from the seas-Current status and microbiological implications[J].Appl Microbiol Biotechnol,2002,59(223):125-134.
[2]陈光英,朱峰,林永成等,南海红树内生真菌1947号次级代谢产物的研究[J].化学研究与应用,2007,19(1):98-99.
[3]朱峰,陈光英,林永成等,南海红树内生真菌编号2526和编号1850代谢产物的研究[J].辽宁师范大学学报(自然科学版),2005,28(3):314-316.
[4]张海龙,田黎,傅红伟等,海洋真菌Alternalia sp.发酵液中化学成分的研究[J].中国中药杂志,2005,30(5):351-353.
[5]李厚金,林永成,刘晓红等,南海海洋真菌Fusarium sp.(#2489)的代谢产物[J].海洋科学,2002,26,(3):57-59.
[6]黄永富,盂娜,田黎等,海洋真菌Aspergillus sp.的二肽类代谢产物的研究[J].中国药物化学杂志,2006,16(1):37-39.
[7]赵文英,朱天骄,古静燕等,海洋来源放线菌3275化学成分及抗肿瘤活性研究[J].天然产物研究与开发,2006,18:405-407.417.
[8]张海龙,傅红伟,田黎等,海洋细菌Bacillus sp.发酵液中化学成分的研究[J].中国海洋药物杂志,2005,24(2):9-12.
[9]赵文英,朱庆书,顾谦群,海洋来源真菌烟曲霉(Sspergillus fuigatus)次级代谢产物研究(Ⅱ)[J].青岛科技大学学报(自然科学版),2007,28,(5):390-393.
[10]陈光英,朱峰,林永成,海洋放线菌Streptomycessp.V5产生的一个新的八元环内酯[J].有机化学,2007,27,(9):1159-1161.
[11]孙奕,吕阿丽,田黎等,海洋真菌Trichoderma reeset的次级代谢产物的分离与鉴定[J].沈阳药科大学学报,2007,24(9):546-548.
[12]朱峰,林永成,周世宁等,南海红树林内源真菌2534号代谢产物的研究[J].中山大学学报(自然科学版),2003,42(1):52-54.
[13]张海龙,田黎,华会明等,海洋真菌Aspergillus versicolor菌丝体中化学成分的研究[J].中国海洋药物杂志,2004,23(4):14-17.
[14]尹文清,张青松,陈柳生等,海洋真菌K26号代谢产物的研究[J].广西师范大学学报(自然科学版),2004,22(3):54-57.
[15]李德海,顾谦群,朱伟明,海洋放线菌11014中抗肿瘤活性成分的研究(Ⅰ)-环二肽[J].中国抗生素杂志,2005,30(8):449-452.
[16]尹宏权,田黎,付红伟等,海洋细菌Bacillus Marinus化学成分分离和结构鉴定[J].北京理工大学学报,2007,27(5):460-462
[17]刘海滨,高昊,王乃利等,红树林真菌草酸青霉(092007)的环二肽类成分[J].沈阳药科大学学报,2007,24(8):474-477.
[18]黄忠京,郭志勇,杨瑞云等,南海红树林内生真菌ZZF42中生物碱类代谢产物的研究[J].中药材,2007,30(8):939-941
[19]Wen Qing YIN,Jie Ming ZOU,Zhi Gang SHE et al,Two Cyclic Peptides Produced by the Endophytic Fungus # 2221from Castaniopsisfissa on the South China Sea Coast[J].Chinese Chemical Letters,2005,16(2):219-222.
[20]尹文清,林永成,周世宁等,南海海洋真菌2516号中的环肽成分[J].中山大学学报(自然科学版),2002,41(4):56-58.
[21]任虹,顾谦群,崔承彬等,海洋来源常现青霉菌SHK1-27生产的蒽醌衍生物及其抗肿瘤活性[J].中国药物化学杂志,2007,17(3):148-153.
[22]朱峰,林永成,周世宁,南海海洋真菌#2526中蒽醌类代谢产物的研究[J].有机化学,2004,24(9):1114-1117.
[23]Wen Han LIN,Hong Zheng FU,Jun LI et al,Novel Chromone Derivatives from Marine Fungus Aspergillus versicolor Isolated from the Sponge Xestospongia exigua[J].Chinese Chemical Letters,2001,12(13):235-238.
[24]Wen Han LIN,Jun LI,Hong Zheng FU et al,Four Novel Hydropyranoindeno Derivatives from Marine Fungus Aspergillus versicolor[J].Chinese Chemical Letters,2001,12(5):435-438.
[25]黄忠京,邵长伦,陈意光等,南海红树林内生真菌ZZF79中吡喃酮类代谢产物的研究[J].中山大学学报(自然科学版),2007,46(4):113-115.
[26]杨瑞云,李春远,林永成等,南海红树林内生真菌Cephalosporium sp.代谢产物[J].应用化学,2007,24(2):220-222.
[27]许志勇,张惠平,海洋真菌09-1-1-1次级代谢产物的研究[J].中国海洋药物杂志,2005,24,(5):43-46.
[28]韩小贤,许晓妍,崔承彬等,海洋真菌烟曲霉H1-04发酵液中的硫代-N哌嗪类产物及其抗肿瘤活性[J].中国药物化学杂志,2007,179(3):155-159.
[29]王军,林永成,吴雄宇等,从南海红树内生真菌No.2533分离出新的异香豆素[J].中山大学学报,2001,4(1):127-128.
[30]张海龙,田黎,华会明等,海洋真菌dspergillus versicolor菌丝体中化学成分的研究[J].中国海洋药物杂志,2004,23(4):14-17.
[31]赵文英,张亚鹏,朱天骄,海洋来源真菌A-f-11中吲哚二酮哌嗪类抗肿瘤活性成分的研究[J].中国抗生素杂志,2006,31(12):749-751.
[32]江红,程元荣,郑卫,海洋碳样小单孢菌产生的大豆黄素和染料木素[J].中国海洋药物杂志,2007,26(1):8-12
[33]陈光英,刘晓红,温露等,南海红树林内生真菌1893代谢产物研究[J].中山大学学报(自然科学版),2003,42(1):49-51.
[34]李厚金,林永成,王立等,南海海洋真菌Hypoxylon oceanicum的代谢产物[J].中山大学学报(自然科学版),2001,40(4):70-72
[35]温露,林永成,赵丽冰等,红树林共生真菌Paecilomyces sp.Tree 1-7代谢产物的研究[J].中国药材,2006,29(8): 782-785
[36]Yongcheng Lin,Zhiyu shao,guangce Jiang,et al.Penicillazine,a unique derivative with 4H-5,6-Dihydro-1,2-oxazine ring system from the marine fungus Penicillium sp.(strain #386)from the south china sea[J].Tetrahedron 2000,56:9607-9609.
[37]Hou Jin LI,Yong Cheng LIN,L.L.P.VRIJMOED,et al,A New Cytotoxic Sterol Produced by an Endophytic Fungus from Castaniopsis fissa at the South China Sea Coast[J].Chinese Chemical Letters,2004,15(4):419-422.
[38]邵志宇,冯永红,邓云霞等,互花米草内生真菌Fusarium sp.F4中的活性代谢物[J].中国天然药物,2007,5(2):109-111.
[39]温露,郭志勇,佘志刚等,一株南海海洋真菌Sporothrix sp.#4335的甾体类代谢产物研究[J].广东药学院学报,2006.22:613-614.
[40]朱峰,彭毓敏,陈光英等,南海海洋真菌2492号中的甾体成分[J].佛山科学技术学院学报(自然科学版),2003,21(6):60-62.
[41]石瑛,田黎,裴月湖,海洋真菌Rhizopus sp.菌丝体中化学成分的研究[J].中国药物化学杂志,2005,15(4):221-223.
[42]宋珊珊,王乃利,高吴等,海洋真菌96F197抗癌活性成分研究[J].中国药物化学杂志,2006,16(2):93-97.
[43]邵志宇,冯永红,邓云霞等,互花米草内生真菌Fusarium sp.F4中的活性代谢物[J].中国天然药物,2007,5(2):109-111.
[44]辛志宏,方玉春,朱天骄等,海绵来源真菌黄灰青霉Sp-19中的抗肿瘤活性成分研究[J].中国海洋药物杂志,2006,25(6):1-15.
[45]Rui Liu,Cheng-Bin Cui,Lin Duan et al,Potent in Vitro Anticancer Activity of Metacycloprodigiosin and Undecylprodigiosin from a Sponge-Derived Actinomycete Saccharopolysporasp[J[.Archives of Pharmacal Research 2005,28(12):1341-1344.
[46]崔洪霞,李富超,阎斌伦等,一株产抑制真菌活性全霉素的胶州湾海洋链霉菌[J].中国海洋药物杂志,2006,25,(1):11-15.
[47]赵文英,朱庆书,顾谦群,海洋来源真菌烟曲霉(Aspergillus fumigatus)次级代谢产物研究(Ⅰ):生物碱[J].青岛科技大学学报(自然科学版),2007,28(3):199-201.
[48]韩小贤,许晓妍,崔承彬等,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[J].中国药物化学杂志,2007,17(4):232-236.
[49]蒋亭,田黎,劳彦斌等,黄海葵附生真菌Penicillium sp.化学成分的研究(Ⅰ)[J].中国海洋药物杂志,2001,20(3):40-42.
[50]李厚金,蔡创华,周毅频等,大亚湾海洋细菌Pseudomonas sp.中的红色素[J].中山大学学报(自然科学版),2003,42(3):102-104.
[51]吴少杰,李富超,秦松等,海洋链霉菌M518产的新星形孢菌素衍生物的生物活性[J].高技术通讯,2007,17(8):874-879.
[52]郑立,韩笑天,陈海敏等,海洋细菌细胞毒代谢产物的分离及其结构解析[J].现代中药研究与实践,2004年,18(增刊):80-83.
[53]温露,袁保红,李厚金等,南海海洋真菌Pseudomonassp.产生的一种抗肿瘤瘤蓝色素[J].中山大学学报(自然科学版),2005,44,(4):63-65.
[54]Xie chao Xiu,Wen Li Mei,You Xing Chao et al,A New Degraded Sesquiterpene from Marine actinomycete streptomyces sp.0616208[J].Chinese Chemical Letters,2006,17(11):1463-1465.
[55]曲新颜,顾谦群,崔承彬等,海洋来源的放线菌3295代谢产物的结构鉴定及抗肿瘤活性(Ⅰ)[J].中国海洋药物杂志,2004,23(6):1-4.
[56]De-Hai Li,Tian-Jiao Zhu,Hong-Bing Liu et al,Four Butenolides are Novel Cytotoxic Compounds Isolated from the Marine-Derived Bacterium,Streptoverticillium luteoverticillatum 11014[J].Archives of Pharmacal Research,2006,29 (8):624-626.
[57]孙奕,吕阿丽,田黎等,海洋真菌Trichoderma reesei的次级代谢产物的分离与鉴定沈阳药科大学学报,2007,24(9):546-548.
[58]乐长高,林永成,南海海洋真菌Julella avicenniae代谢产物的研究(Ⅱ)[J].中草药,2006,37(1):16-18
[59]李春远,杨瑞云,林永成,海洋真菌K38号代谢产物的研究[J].中山大学学报(自然科学版),2007,46(1):67-70.
[60]Yongcheng Lin,Xiongyu Wu,Shuang Feng,et al.Five unique compounds:xyloketal from mangrove fungus Xylaria sp.from the South China Sea Coast[J].J org chem,2001,66,6252-6256.
[2]潘启超,胥彬,肿瘤药理学与化学治疗学[M].第二版,郑州:河南医科大学出版社,3-10
[3]Proksch P.,Edrada R.A.,Eber.1..Drugs from the seas-Current status and microbiological implications[J].Appl Microbiol Biotechnol,2002,59(223):125-134
[4]刘景生.细胞信息与调控.现代生物医学丛书.北京:北京医科大学、北京协和医科大学联合出版社.1998:318-333
[5]Keer JF,Wyllie AH,Currie AR,Apoptosis,et al,a basic biological phenomenon with wide-ranging implications in tissue kinetics,Br J Cancer.1972,26:239-251
[6]王海燕,蔡兵,崔承彬,等.蔓荆子活性成分vitexicarpin诱导K562细胞凋亡的机制[J].药学学报,2005,40(1):27-31.
[7]Nikaido H.,Prevention of drug acessto bacterial tagets:permeability barriers and active effiux[J].Science,1994,264:382
[8]Spratt B G.,Resistance to antibiotics mediated by target alerration[J].Science,1994,264:388
[9]沈同,王镜岩.生物化学(下册)北京:高等教育出版社2000,398
[10]Finken,M.,P.Kirschner,A.Meier,A.Wrede,and E.C.Bo"ttger.Molecular basis of streptomycin resistance in Mycobacterium tuberculosis:alterations of the ribosomal protein S12 gene and point mutations within a functional 16S ribosomal RNA pseudoknot.Mol.Microbiol.1993.9:1239-1246.
[11]Cundlife E.Recognition sites for antibiotics with in rRNA.In:Hill W E,Dahlberg A,Carret RA,et al.The ribosome:structure,function and evolution.[M].Washington DC:American Society for Microbiology,1990:479
[12]Hima J,Hesketh A,Okamato S,et al.Induction of actinorhodin production by rpsl(encoding ribosomal protein S12)mutation that confer streptomycin resistance in Streptomyces lividans and Streptomyces coelicolor A3(2)[J].Bacteriol,1996,178(24):7276-7284
[13]Hu.H.F,Ochi K.Novel approach for improving the productivity of antibiotic-producing strains by inducing combined resistant mutations.Appl Environ Microbiol,2001,67(4):1885-1892
[14]Hosoya Y,Okamoto S,Muramatsu H et al.Acquisition of certain streptomycin-resistant(str)mutations enhances antibiotic production in bacteria.Antimicrob.Agents.Chemotherapy,1998,42(8):2941-2047
[15]涂国全等.通过获得链霉素抗性基因(str)突变株筛选梅岭霉素高产菌株[J].中国抗生素杂志,2002,27(6):321-325
[16]陶纯长等.链霉素抗性突变理性筛选avermetin高产菌株.中国抗生素杂志[J].2002,27(9):521-523
[17]杨东靖,陈冠群等.纳他霉素高产菌株的链霉素抗性选育及其发酵工艺的优化[J].药物生物技术.2003,10(2):84-87
[18]Yoshiko O H,Susumu O,and Kozo Ochi,Development of Antibiotic-Overproducing Strains by Site-Directed Mutagenesis of the rpsL Gene in Streptomyces lividans,Appl and Environment Microbiology,July 2003,4256-4259
[19]Hopwood D A,Chater K F.Fresh approaches to antibiotic production[M].Philos Trans R Soc Lond B Biol Sci,1980,290(1040):313
[20]顾觉奋,获取抗生素的新途径—沉默基因的激活,国外医药抗生素分册[J].2002,24(5):193-197
[21]Hopwood D A.Towards on understanding of gene switching in Streptomyces,the basis of sporulation and antibiotics production[M]Proc R Soc Lond Biol,1988,235:121
[22]崔承彬,顾谦群等.开拓微生物药用资源暂新领域的新技术方法学研究.第八届中国海洋药物学术研讨会.青岛.2002.45-47
[23]王海燕,蔡兵,崔承彬,等.蔓荆子活性成分vitexicarpin诱导K562细胞凋亡的机制[J].药学学报,2005,40(1):27-31.
[24]连文琰.中国防己科药用植物的初步调查[J].植物分类学报,1975,13(1):32-53.
[25]冉先德.中华药海[M].第一版,哈尔滨:哈尔滨出版社,1993:1925.
[26]梁培瑜,周琦,周法兴.细圆藤的化学成分研究[J].中国中药杂志,1998,23(1):39-40.
[27]李长伟.南酸枣抗肿瘤活性成分的研究[M].沈阳药科大学硕士学位论文,2005:13-17.
[28]CHATURVEDULA V S P,SCHILLING J K,MILLER J S,et al.Two new triterpene esters from the twigs of Brachylaena ramifiora from the Madagascar rainforest[J].J Nat Prod,2002,65(8):1222-1224.
[29]MATUSUNAGA S,MORIITA R.Hopenol-B,A triterpene alcohol from Euphorbia supina[J].Phytochemistry,1983,22(2):605-606.
[30]MCLEAN S,PERPICN-DUMONT M,REYNOLDS W F,et al.Unambiguous structural and nuclear magnetic resonance spectral characterization of two triterpenoids of Maprounea guiqnensis by two-dimensional nuclear magnetic resonance spectroscopy[J].Can J Chem,1987,65(11): 2519-2525.
[3l]TSTUDA Y,HATORI M.Total synthesis of hydroxyhopanone[J].Chem Pharm Bull,1967,15(7):1073-1074.
[32]ROWAN D D,RUSSELL G B.3β-merthoxyhop-22(29)-ene from Chionochloa cheesemanii[J].Phytochemistry,1992,31(2):702-703.
[33]WILKINS A L,RONALDSON K J,JAGER P M,et al.A ~(13)C NMR Study of some oxygenated hopane triterpenes[J].Aust J Chem,1987,40(10):1713-1721.
[34]NAKANO T,HASEGAWA M.Isolation of erythrodiol 3-palmitate from Bumelia obtusifolia[J].Planta Med,1975,27(1):89-91.
[35]WANDJI J,TILLEQUIN F,MULHOLLAND D A,et al.Fatty acid esters of triterpenoids and steroid glycosides from Gambeya africana[J].Planta Med,2002,68(9):822-826.
[36]CONNOLLY J D,FREER A A.Structure of 5β,24-cyclofriedelan-3-one,a novel hexacyclic triterpenoid in space group P1[J].Acta Cryst,1986,C42:1352-1354.
[37]朱峰,林永成,周世宁等,南海红树林内源真菌2534号代谢产物的研究[J].中山大学学报(自然科学版)2003,42(1):52-54
[38]江红,程元荣,郑卫,海洋碳样小单孢菌产生的大豆黄素和染料木素[J].中国海洋药物杂志2007,26(1):8-12
[39]张海龙,田黎,傅红伟等,海洋真菌Alternalia sp.发酵液中化学成分的研究[J].中国中药杂志2005,30(5):351-353
[40]Romer A.~1H NMR spectra of substituted phenazines[J].Org Mag Res,1982,19(2):66-68.
[41]Breitmaier E,Hollstein U.Carbon-13 Nuclear magnetic resonance chemical shifts of substituted phenazines[J].J Org Chem.1976,41(12):2104-2108.
[1]P.Proksch,R.A.Edrada,r.Ebel.Drugs from the seas-Current status and microbiological implications[J].Appl Microbiol Biotechnol,2002,59(223):125-134.
[2]陈光英,朱峰,林永成等,南海红树内生真菌1947号次级代谢产物的研究[J].化学研究与应用,2007,19(1):98-99.
[3]朱峰,陈光英,林永成等,南海红树内生真菌编号2526和编号1850代谢产物的研究[J].辽宁师范大学学报(自然科学版),2005,28(3):314-316.
[4]张海龙,田黎,傅红伟等,海洋真菌Alternalia sp.发酵液中化学成分的研究[J].中国中药杂志,2005,30(5):351-353.
[5]李厚金,林永成,刘晓红等,南海海洋真菌Fusarium sp.(#2489)的代谢产物[J].海洋科学,2002,26,(3):57-59.
[6]黄永富,盂娜,田黎等,海洋真菌Aspergillus sp.的二肽类代谢产物的研究[J].中国药物化学杂志,2006,16(1):37-39.
[7]赵文英,朱天骄,古静燕等,海洋来源放线菌3275化学成分及抗肿瘤活性研究[J].天然产物研究与开发,2006,18:405-407.417.
[8]张海龙,傅红伟,田黎等,海洋细菌Bacillus sp.发酵液中化学成分的研究[J].中国海洋药物杂志,2005,24(2):9-12.
[9]赵文英,朱庆书,顾谦群,海洋来源真菌烟曲霉(Sspergillus fuigatus)次级代谢产物研究(Ⅱ)[J].青岛科技大学学报(自然科学版),2007,28,(5):390-393.
[10]陈光英,朱峰,林永成,海洋放线菌Streptomycessp.V5产生的一个新的八元环内酯[J].有机化学,2007,27,(9):1159-1161.
[11]孙奕,吕阿丽,田黎等,海洋真菌Trichoderma reeset的次级代谢产物的分离与鉴定[J].沈阳药科大学学报,2007,24(9):546-548.
[12]朱峰,林永成,周世宁等,南海红树林内源真菌2534号代谢产物的研究[J].中山大学学报(自然科学版),2003,42(1):52-54.
[13]张海龙,田黎,华会明等,海洋真菌Aspergillus versicolor菌丝体中化学成分的研究[J].中国海洋药物杂志,2004,23(4):14-17.
[14]尹文清,张青松,陈柳生等,海洋真菌K26号代谢产物的研究[J].广西师范大学学报(自然科学版),2004,22(3):54-57.
[15]李德海,顾谦群,朱伟明,海洋放线菌11014中抗肿瘤活性成分的研究(Ⅰ)-环二肽[J].中国抗生素杂志,2005,30(8):449-452.
[16]尹宏权,田黎,付红伟等,海洋细菌Bacillus Marinus化学成分分离和结构鉴定[J].北京理工大学学报,2007,27(5):460-462
[17]刘海滨,高昊,王乃利等,红树林真菌草酸青霉(092007)的环二肽类成分[J].沈阳药科大学学报,2007,24(8):474-477.
[18]黄忠京,郭志勇,杨瑞云等,南海红树林内生真菌ZZF42中生物碱类代谢产物的研究[J].中药材,2007,30(8):939-941
[19]Wen Qing YIN,Jie Ming ZOU,Zhi Gang SHE et al,Two Cyclic Peptides Produced by the Endophytic Fungus # 2221from Castaniopsisfissa on the South China Sea Coast[J].Chinese Chemical Letters,2005,16(2):219-222.
[20]尹文清,林永成,周世宁等,南海海洋真菌2516号中的环肽成分[J].中山大学学报(自然科学版),2002,41(4):56-58.
[21]任虹,顾谦群,崔承彬等,海洋来源常现青霉菌SHK1-27生产的蒽醌衍生物及其抗肿瘤活性[J].中国药物化学杂志,2007,17(3):148-153.
[22]朱峰,林永成,周世宁,南海海洋真菌#2526中蒽醌类代谢产物的研究[J].有机化学,2004,24(9):1114-1117.
[23]Wen Han LIN,Hong Zheng FU,Jun LI et al,Novel Chromone Derivatives from Marine Fungus Aspergillus versicolor Isolated from the Sponge Xestospongia exigua[J].Chinese Chemical Letters,2001,12(13):235-238.
[24]Wen Han LIN,Jun LI,Hong Zheng FU et al,Four Novel Hydropyranoindeno Derivatives from Marine Fungus Aspergillus versicolor[J].Chinese Chemical Letters,2001,12(5):435-438.
[25]黄忠京,邵长伦,陈意光等,南海红树林内生真菌ZZF79中吡喃酮类代谢产物的研究[J].中山大学学报(自然科学版),2007,46(4):113-115.
[26]杨瑞云,李春远,林永成等,南海红树林内生真菌Cephalosporium sp.代谢产物[J].应用化学,2007,24(2):220-222.
[27]许志勇,张惠平,海洋真菌09-1-1-1次级代谢产物的研究[J].中国海洋药物杂志,2005,24,(5):43-46.
[28]韩小贤,许晓妍,崔承彬等,海洋真菌烟曲霉H1-04发酵液中的硫代-N哌嗪类产物及其抗肿瘤活性[J].中国药物化学杂志,2007,179(3):155-159.
[29]王军,林永成,吴雄宇等,从南海红树内生真菌No.2533分离出新的异香豆素[J].中山大学学报,2001,4(1):127-128.
[30]张海龙,田黎,华会明等,海洋真菌dspergillus versicolor菌丝体中化学成分的研究[J].中国海洋药物杂志,2004,23(4):14-17.
[31]赵文英,张亚鹏,朱天骄,海洋来源真菌A-f-11中吲哚二酮哌嗪类抗肿瘤活性成分的研究[J].中国抗生素杂志,2006,31(12):749-751.
[32]江红,程元荣,郑卫,海洋碳样小单孢菌产生的大豆黄素和染料木素[J].中国海洋药物杂志,2007,26(1):8-12
[33]陈光英,刘晓红,温露等,南海红树林内生真菌1893代谢产物研究[J].中山大学学报(自然科学版),2003,42(1):49-51.
[34]李厚金,林永成,王立等,南海海洋真菌Hypoxylon oceanicum的代谢产物[J].中山大学学报(自然科学版),2001,40(4):70-72
[35]温露,林永成,赵丽冰等,红树林共生真菌Paecilomyces sp.Tree 1-7代谢产物的研究[J].中国药材,2006,29(8): 782-785
[36]Yongcheng Lin,Zhiyu shao,guangce Jiang,et al.Penicillazine,a unique derivative with 4H-5,6-Dihydro-1,2-oxazine ring system from the marine fungus Penicillium sp.(strain #386)from the south china sea[J].Tetrahedron 2000,56:9607-9609.
[37]Hou Jin LI,Yong Cheng LIN,L.L.P.VRIJMOED,et al,A New Cytotoxic Sterol Produced by an Endophytic Fungus from Castaniopsis fissa at the South China Sea Coast[J].Chinese Chemical Letters,2004,15(4):419-422.
[38]邵志宇,冯永红,邓云霞等,互花米草内生真菌Fusarium sp.F4中的活性代谢物[J].中国天然药物,2007,5(2):109-111.
[39]温露,郭志勇,佘志刚等,一株南海海洋真菌Sporothrix sp.#4335的甾体类代谢产物研究[J].广东药学院学报,2006.22:613-614.
[40]朱峰,彭毓敏,陈光英等,南海海洋真菌2492号中的甾体成分[J].佛山科学技术学院学报(自然科学版),2003,21(6):60-62.
[41]石瑛,田黎,裴月湖,海洋真菌Rhizopus sp.菌丝体中化学成分的研究[J].中国药物化学杂志,2005,15(4):221-223.
[42]宋珊珊,王乃利,高吴等,海洋真菌96F197抗癌活性成分研究[J].中国药物化学杂志,2006,16(2):93-97.
[43]邵志宇,冯永红,邓云霞等,互花米草内生真菌Fusarium sp.F4中的活性代谢物[J].中国天然药物,2007,5(2):109-111.
[44]辛志宏,方玉春,朱天骄等,海绵来源真菌黄灰青霉Sp-19中的抗肿瘤活性成分研究[J].中国海洋药物杂志,2006,25(6):1-15.
[45]Rui Liu,Cheng-Bin Cui,Lin Duan et al,Potent in Vitro Anticancer Activity of Metacycloprodigiosin and Undecylprodigiosin from a Sponge-Derived Actinomycete Saccharopolysporasp[J[.Archives of Pharmacal Research 2005,28(12):1341-1344.
[46]崔洪霞,李富超,阎斌伦等,一株产抑制真菌活性全霉素的胶州湾海洋链霉菌[J].中国海洋药物杂志,2006,25,(1):11-15.
[47]赵文英,朱庆书,顾谦群,海洋来源真菌烟曲霉(Aspergillus fumigatus)次级代谢产物研究(Ⅰ):生物碱[J].青岛科技大学学报(自然科学版),2007,28(3):199-201.
[48]韩小贤,许晓妍,崔承彬等,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[J].中国药物化学杂志,2007,17(4):232-236.
[49]蒋亭,田黎,劳彦斌等,黄海葵附生真菌Penicillium sp.化学成分的研究(Ⅰ)[J].中国海洋药物杂志,2001,20(3):40-42.
[50]李厚金,蔡创华,周毅频等,大亚湾海洋细菌Pseudomonas sp.中的红色素[J].中山大学学报(自然科学版),2003,42(3):102-104.
[51]吴少杰,李富超,秦松等,海洋链霉菌M518产的新星形孢菌素衍生物的生物活性[J].高技术通讯,2007,17(8):874-879.
[52]郑立,韩笑天,陈海敏等,海洋细菌细胞毒代谢产物的分离及其结构解析[J].现代中药研究与实践,2004年,18(增刊):80-83.
[53]温露,袁保红,李厚金等,南海海洋真菌Pseudomonassp.产生的一种抗肿瘤瘤蓝色素[J].中山大学学报(自然科学版),2005,44,(4):63-65.
[54]Xie chao Xiu,Wen Li Mei,You Xing Chao et al,A New Degraded Sesquiterpene from Marine actinomycete streptomyces sp.0616208[J].Chinese Chemical Letters,2006,17(11):1463-1465.
[55]曲新颜,顾谦群,崔承彬等,海洋来源的放线菌3295代谢产物的结构鉴定及抗肿瘤活性(Ⅰ)[J].中国海洋药物杂志,2004,23(6):1-4.
[56]De-Hai Li,Tian-Jiao Zhu,Hong-Bing Liu et al,Four Butenolides are Novel Cytotoxic Compounds Isolated from the Marine-Derived Bacterium,Streptoverticillium luteoverticillatum 11014[J].Archives of Pharmacal Research,2006,29 (8):624-626.
[57]孙奕,吕阿丽,田黎等,海洋真菌Trichoderma reesei的次级代谢产物的分离与鉴定沈阳药科大学学报,2007,24(9):546-548.
[58]乐长高,林永成,南海海洋真菌Julella avicenniae代谢产物的研究(Ⅱ)[J].中草药,2006,37(1):16-18
[59]李春远,杨瑞云,林永成,海洋真菌K38号代谢产物的研究[J].中山大学学报(自然科学版),2007,46(1):67-70.
[60]Yongcheng Lin,Xiongyu Wu,Shuang Feng,et al.Five unique compounds:xyloketal from mangrove fungus Xylaria sp.from the South China Sea Coast[J].J org chem,2001,66,6252-6256.