青霉HLKG-44次级代谢产物研究及膦二肽的合成与质谱研究
|
摘要
天然产物具有广泛的生物活性,是人们获得药物的主要途径之一。微生物广泛地分布于整个生物圈,具有丰富多样的代谢产物,是天然药物的一个重要的来源。本论文以沙溪河污染地分离得到的一株具有抗肿瘤活性的青霉菌作为研究菌株,希望通过对其发酵、分离得到具有抗肿瘤活性的次级代谢产物。
我们对青霉菌HLKG-44进行发酵培养并采用不同的分离方法对其脂溶性次级代谢产物进行初步分离分析,分离出6个化合物,进一步通过核磁(~1H NMR、~(13)CNMR、DEPT、HSQC、HMBC等)、质谱、单晶衍射、红外和元素分析等方法确定化合物的结构。对化合物进行了抗肺肿瘤细胞A549的活性测试,结果显示其中的两个化合物具有比较好的抗肿瘤活性,半数抑制浓度(IC_(50))分别是0.02μg/mL和18.9μg/mL。
磷不仅在生命代谢和遗传过程中起着重要的作用,而且在生命起源、发育以及所有生命化学过程中都扮演重要的角色。磷化学与生命活动有着非常密切的关系。研究表明,膦肽作为一类含磷化合物具有抑制植物生长、抑制癌细胞生长、抗烟草花叶病毒的活性,同时也是丝氨酸蛋白酶类(如胰蛋白酶、胰凝乳蛋白酶、凝血酶、粒酶、二肽基肽酶等)的一类重要的抑制剂。这些酶在体内代谢中具有重要作用,通过这类化合物的研究,可为药物开发建立基础。
本文探索出产率好、成本低、简单有效的无溶剂合成膦二肽前体氨基亚膦酸酯(APP)的方法。合成了5个新型膦二肽化合物,并对其进行结构鉴定。通过磷谱、HPLC、晶体衍射及圆二色分析方法对膦二肽合成过程中的氨基酸选择性与APP反应进行研究,表明Trp和Pro对消旋的(R/S)-APP具有完全的选择性,Ile、Val及Phe只具有部分的选择性。对膦二肽ESI-MS的裂解规律进行研究,发现和验证其两个特殊的重排。以胃蛋白酶为模型研究其抑制丝氨酸蛋白酶的活性,表明该类化合物并没有抑制活性,对其他类丝氨酸蛋白酶的抑制作用有待于进一步研究。
Natural medicines are the primary access to drugs because of the wide range of biological activity.Microbes,being widely distributed throughout the biosphere,are important natural medicine sources for their rich and varied metabolic products.In this paper,a kind of penicillin with anti-tumor activity,which is isolated from the polluted area of Shaxi River,is studied.It is expected to obtain the secondary metabolites with anti-tumor activity through fermentation and separation from the penicillin HLKG-44.
After fermentation the penicillin HLKG-44,we isolate and analyze its lipophilic constituents.Six conpounds were isolated by different isolation methods,then identification their chemical structure by NMR(~1H NMR,~(13)C NMR,DEPT,HSQC, HMBC,etc.),MS,X-ray diffraction,infra-red,elemental analysis and other methods. The result of the anti-lung tumor cells A549 test showed that two of the compounds have better anti-tumor activity,50%inhibitory concentration(IC_(50)) were 0.02μg/mL and 18.9μg/mL respectively.
Phosphorus play an important role not only in the metabolic and genetic process, but also in the origin,growth and all other chemical process of life.Previous study shows that phosphono-peptides,as one type of phospho compounds,can inhibit plant growth,the growth of cancer cells,anti-tobacco mosaic virus activity,and also a major inhibitor of serine protease(such as trypsin,chynotrupsin,thrombin,granzyme,uPA, and so on).These enzymes play an important role in the metabolism.Based on the study of such compounds,it is expected to provide an important basis for the drug development.
A high production,low cost,simple and effective solvent-free synthesis method to obtain the phosphonic dipeptide precursor amino-phosphonate(APP) is proposed in this paper.Five kinds of new phospho-dipeptides are synthesized,their structure are identified.Through the spectrum of phosphorus(~(31)P NMR),HPLC,crystal diffraction and circular dichroism analysis,selective of amino acids and APP reaction ability in the synthesis process is studied.The results showed that trptophane and proline show selectivity for one of the optical isomer of APP,while isoleucine,valine and phenylalanine show only partial selectivity.The fragment rule of ESI-MS for phospho-dipeptides was studied and two special rearrangements were werified.Research of the serine proteinase activity based on the model of pepsin showed that these compounds do not have significant inhibition activity.Inhibition of other types of serine protease will be studied further.
我们对青霉菌HLKG-44进行发酵培养并采用不同的分离方法对其脂溶性次级代谢产物进行初步分离分析,分离出6个化合物,进一步通过核磁(~1H NMR、~(13)CNMR、DEPT、HSQC、HMBC等)、质谱、单晶衍射、红外和元素分析等方法确定化合物的结构。对化合物进行了抗肺肿瘤细胞A549的活性测试,结果显示其中的两个化合物具有比较好的抗肿瘤活性,半数抑制浓度(IC_(50))分别是0.02μg/mL和18.9μg/mL。
磷不仅在生命代谢和遗传过程中起着重要的作用,而且在生命起源、发育以及所有生命化学过程中都扮演重要的角色。磷化学与生命活动有着非常密切的关系。研究表明,膦肽作为一类含磷化合物具有抑制植物生长、抑制癌细胞生长、抗烟草花叶病毒的活性,同时也是丝氨酸蛋白酶类(如胰蛋白酶、胰凝乳蛋白酶、凝血酶、粒酶、二肽基肽酶等)的一类重要的抑制剂。这些酶在体内代谢中具有重要作用,通过这类化合物的研究,可为药物开发建立基础。
本文探索出产率好、成本低、简单有效的无溶剂合成膦二肽前体氨基亚膦酸酯(APP)的方法。合成了5个新型膦二肽化合物,并对其进行结构鉴定。通过磷谱、HPLC、晶体衍射及圆二色分析方法对膦二肽合成过程中的氨基酸选择性与APP反应进行研究,表明Trp和Pro对消旋的(R/S)-APP具有完全的选择性,Ile、Val及Phe只具有部分的选择性。对膦二肽ESI-MS的裂解规律进行研究,发现和验证其两个特殊的重排。以胃蛋白酶为模型研究其抑制丝氨酸蛋白酶的活性,表明该类化合物并没有抑制活性,对其他类丝氨酸蛋白酶的抑制作用有待于进一步研究。
Natural medicines are the primary access to drugs because of the wide range of biological activity.Microbes,being widely distributed throughout the biosphere,are important natural medicine sources for their rich and varied metabolic products.In this paper,a kind of penicillin with anti-tumor activity,which is isolated from the polluted area of Shaxi River,is studied.It is expected to obtain the secondary metabolites with anti-tumor activity through fermentation and separation from the penicillin HLKG-44.
After fermentation the penicillin HLKG-44,we isolate and analyze its lipophilic constituents.Six conpounds were isolated by different isolation methods,then identification their chemical structure by NMR(~1H NMR,~(13)C NMR,DEPT,HSQC, HMBC,etc.),MS,X-ray diffraction,infra-red,elemental analysis and other methods. The result of the anti-lung tumor cells A549 test showed that two of the compounds have better anti-tumor activity,50%inhibitory concentration(IC_(50)) were 0.02μg/mL and 18.9μg/mL respectively.
Phosphorus play an important role not only in the metabolic and genetic process, but also in the origin,growth and all other chemical process of life.Previous study shows that phosphono-peptides,as one type of phospho compounds,can inhibit plant growth,the growth of cancer cells,anti-tobacco mosaic virus activity,and also a major inhibitor of serine protease(such as trypsin,chynotrupsin,thrombin,granzyme,uPA, and so on).These enzymes play an important role in the metabolism.Based on the study of such compounds,it is expected to provide an important basis for the drug development.
A high production,low cost,simple and effective solvent-free synthesis method to obtain the phosphonic dipeptide precursor amino-phosphonate(APP) is proposed in this paper.Five kinds of new phospho-dipeptides are synthesized,their structure are identified.Through the spectrum of phosphorus(~(31)P NMR),HPLC,crystal diffraction and circular dichroism analysis,selective of amino acids and APP reaction ability in the synthesis process is studied.The results showed that trptophane and proline show selectivity for one of the optical isomer of APP,while isoleucine,valine and phenylalanine show only partial selectivity.The fragment rule of ESI-MS for phospho-dipeptides was studied and two special rearrangements were werified.Research of the serine proteinase activity based on the model of pepsin showed that these compounds do not have significant inhibition activity.Inhibition of other types of serine protease will be studied further.
引文
[1]Oberlies N H,Kroll D J.J.Nat.Prod,2004,67:129.
[2]Fitzpatrick F A,Wheeler R.Int.Immunopharmacol,2003,3:1699.
[3]Newraan D J,Cragg G M,Snader K M.J.Nar.Prod,2003,55:1022.
[4]刘成梅,游海.天然产物有效成分的分离与应用.化学工业出版社.2003.
[5]沈萍.微生物学.北京:高等教育出版社.2000.
[6]黄秀梨.微生物学.北京:高等教育出版社.2003.
[7]周德庆.微生物学教程.北京:高等教育出版社.2002.
[8]Account to:Editors,Antifungal Antibiotics[J].J Antibiot,1996-2005,49-58:1.
[9]张致平.中国处方药,2006,48(3):28.
[10]Sanflard D.Curr Opin Microbiol,2002,5(4):379.
[11]李端等.药理学(第四版).人民卫生出版社.2000.
[12]杨宝峰.药理学(五年制教材).北京医科大学出版社.2003.
[13]李全,谢毓敏.药学进展,1996,20(3):129.
[14]Tremont.Lukats.W,Gilbert MR.Cancer Control,2003,10(2):125.
[15]方美娟.厦门大学傅士毕业论文.2006.
[16]Atalay G,Candoso F,Awada A,Et al.Ann Oncol,2003,14(9):1346.
[17]Singh S B,Lingham R B.Pharmacol Ther,1994,62(1-2):57.
[18]Osterhage C.Dissertation of Technischen Universitat Carolo-Wilhelmina.2001.
[19]Fleming A Br.J.Exp.Pathol,1929,10:226.
[20]Dreyfuss M,Harri E,Hotmann H,Kobel H,Pache W,Tscherter H.Cyclosporin A and C.Eur.J.Appl.Microbiol.Biotechnol,1976,3:125.
[21]Goodman G A,Goodman L S,Rall TW,Murad F.(eds.) Macmillan.7th ed.New York.1985,1298.
[22]Rehm H J,etc.Industrielle Mikrobiologie.2nd ed.Berlin:Springer-Verlag,1980.
[23]Alberts A W,Chen J,Kuron G,etc.Proc.Natl.Acad.Sci.U.S.A,1980,77(7):3957.
[24]于能江,郭顺星.中国药学杂志.1999,34(9):577.
[25]Romila D C,Gerhard S,Jeffrey J,Valerie B,Feng X D,Guy T C.Diazepinomicin.J Nat Prod,2004,67:1431.
[26]Kerstin Liberra and Ulrike Lindequist.Pharmazie,1995,50(9):583.
[27]Ring-Chung cheng,Mustafa varoglu,Leif abrell,etc,J.Org.chem,1994,59:6344.
[28]Atsushi Numata,Chika Takahashi,Yoshinod Ito,etc.Tetrahedron left,1992,33(12):1621.
[29]Wei H,Itoh T,Kinoshita M,Nakai Y,etc.Tetrahedron,2004,60:6015.
[30]Christine K,Stefan K,Anthony D W,Gabriele M K.J Nat Prod,2004,67:1058.
[31]Cueto M,MacMillan JB,Jensen PR,Fenical W.Phytochemistry,2006,67(16):1826.
[32]Jeffrey T G,Taro A,Akiko A,etc.J Nat Prod,2004,67:362.
[33]Varoglu M,Crews P,etc.J.Nat.Prod,2000,63(1):41.
[34]Lee S M,Li X F,Jiang H L,Cheng JG;etc.Tetrahedron Lett,2003,44(42):7707.
[35]Emsley J,Hall D.The chemistry of Phosphorus.London:Harper & Row,1976.
[36]赵玉芬,赵国辉,麻远.磷与生命科学.清华大学出版社,2005.
[37]Chen R-Y,Mao L-J.Chem.J.Chin.Univ.1995,16:892(in Chinese).
[38]陈何如,徐杰诚.药学学报.1996,31(9):671.
[39]李在国,黄润秋,杨昭,邵瑞链.应用化学,1999,02:94.
[40]Wang Q M,Li Z G,Huang R Q,Cheng J R.Heteroat.Chem.2001,12:68.
[41]Liu X J,Chen R Y,Yang Y Y.Chem.J.Chin.Univ.2002,23:1299(in Chinese).
[42]Kristel Senten,Liesbeth Daniels,Pieter Van der Veken,etc.J.Comb.Chem.2003,5:336.
[43]J Joossens,P Vander Veken,A M,etc.J.Med.Chem.2004,47:2411.
[44]Yuan C Y,Wang G H,etc.Synthesis,1990,3:256.
[45]Shi D Q,Chen R.Y.Chin.J.Appl.Chem.2002,19:780(in Chinese).
[46]Seyferth D,Marmor R S,Hilbert P.J.Org.Chem.1971,36(10):1379.
[47]Shono T,Matsumura Y,Tsubata K.Tetra.Letts.1981,22(34):3249.
[48]Natchev I A,etc.Tetrahedron 1988;44(20):6465.
[49]Kudzin Z H,Kotynsi A.Synthesis,1980,1028.
[50]Asano S,Kitahara T,Ogawa T,et al.Agr Biol.Chem,1973,37:193.
[51]Darrell K,Berlin N K,Roy R T,et al.Synthesis,1968,4494.
[52]Darrell K,Berlin N K,Claunch R T,et al.J.Org.Chem,1968,33:3090.
[53]Green D,Elgendy S,Baban J.A,et al.Tetrahedron Letts,1993,34:6917.
[54]Lejczak B,Kafarski P,Mokokowiecka E.Biochem.J,1987,242:81.
[55]Rachon J,Schollkopf U,Wintel T H.Lieb.Ann.Chem,1981,709.
[56]Chalmers M.E,Kosolapoff G.M.J.Am.Chem.Soc.1953,75:5278.
[57]Tyka R,etc.Tetra.Letts.1970,9:677.
[58]Oleksyszyn J,Gruszecka E,etc.Tetra Letts.1981,22(36):3537.
[59]Oleksyszyn J,Tyka R,etc.Tetra.Letts.1977,32:2823.
[60]Kudzin Z H,Stec W J,etc.Synthesis,1978,6:469.
[61]Gajda T,Matusiak M,etc.Syn.Commun,1992,22:2193.
[62]Lukszo J,Tyka R,etc.Synthesis,1977,239.
[63]Green D,Patel G,Elgendy S,et al.Tetrahedron,1994,50:5099.
[64]Gilmore W F,Mcbride HA.J.Am.Chem.Soc.1972,94:4361.
[65]Amos B.Smith,Kraig M.Yager,and Carol M.Taylor.J.Am.Chem.Soc.1995,117:10879.
[66]宁永成.有机化合物结构鉴定与有机波谱学(第二版),北京:科学出版社,2000.
[67]唐恢同.有机化合物光谱鉴定,北京大学出版社,1994.
[68]Burlingame A L,Boyd R K,Gaskell S.T,Anal,Chem,1996,68:599R.
[69]Matsuo T,Seyame Y,J.Mass.Spectrom,2000,35:114.
[70]Wigger Mafia,Eyler John R,etc.J.Am.Soc.Mass Spectrom,2002,13:1162.
[71]Davis G D,etc.Jr.ACC.Chem.Res,1979,12:359.
[72]Munson B,Field F H,etc.J.Am.Chem.Soc,1966,88:2621.
[73]Barber M,Bordoli R S,Scdgwick R D.et al,Nature,1981,293:270.
[74]Fenn J B,Mann M,Meng C K,et al,Science,1989,246:64.
[75]Smith R D,Loo J A,et al,Anal Chem,1990,62:882.
[76]ReidN W,etc.Adv.Mass Spectrom,1980,8:1480.
[77]Karas M,Hillenkamp F,etc.Anal,Chem,1988,60:2299-2301.
[78]Beavis R C,Chait B T,etc.Rapid Commun.Mass Spectrom,1989,3:233.
[79]Hillenkamp F,Karas M,Beavis R C,Anal Chem,1991,63:1193A.
[80]Fenselau C,etc.Anal Chem,1997,69:661A.
[81]Biemann K,etc.Annu.Rev.Biochem,1992,61:977.
[82]Beavis R.C.et al,Anal,Chem,1990,62:1836.
[83]McLafferty F W,etc.J.Am.Soc.Mass Spectrom,1997,8:380.
[84]Sheldrick G M.(1997).SHELXS-97 and SHELXL97.University of Gottingen,Germany.
[85]Accelrys(2001).ViewerPro(V4.2).Accelrys Inc,Burlington,Massachusetts,USA.
[86]Bruker(2001).SAINT(V6.22),SMART(V5.625) and SADABS(V2.03).Bruker AXS Inc,Madison,Wisconsin,USA.
[87]Farrugia L J,etc.J.Appl.Cryst.1997,30:565.
[88]AN Starratt.Phytochem,EN,1976,15:2002.
[89]C.Delseth,Helv.Chim.Acta,EN,1979,62:2037.
[90]Lan D Entwistlf,Colin C Howard,etc.Phytochemstry 1974,13:173.
[91]Singleon V L,Bohonos N,Ullstrup A J,Decumbin.Nature,1958,181:1072.
[92]Betina V,Nemec P,Dobias J,Barath A,Cyanein.Folia Microbiol.1962,7:353.
[93]Suzuki Y,Tanaka H,Aoki H,Tamura T.Agric.Biol.Chem.1979,34:395.
[94]Zhu J W,Hori H,Tsukuda T,etc.Bioorg.Med.Chem.Lett.1997,7:139.
[95]Ahmaol,et al.[J].PYTCAS;Phytochemistry 1976,15:2000.
[96]Delseth,C.et al.HCACAV,Helv.Chim.Acta;EN.1979,62:2037.
[97]Schmidt Bernd;Nave Atefan.Advanced Synthesis & Catalysis.2007,349(1+2):215.
[98]Takahashi H,Yoshioka M,Imai N.Synthesis.1994,6:763.
[99]Takahashi H,Yoshioka M,Imai N.Synthesis 1994,6:763.
[100]Takahiko Akiyama,MachiKo Sanada,Kohei Fuchibe.Synlett 2003,10:1463.
[101]Babak Kaboudin,Rahman Nazari,etc.Tetrahedron Letters.2001,42:8211.
[102]Babak Kaboudin and Khavar Moradia.Tetrahedron Letters.2005,46(17):2989.
[103]方华.厦门大学博士毕业论文.2007.
[104]Hamilton R,Walker B,etc.Tetrahedron Lett.1993,34(17):2847.
[105]Wang C J,Taylor T L,Mical A J,etc.Tetrahedron Lett.1992,33(50):7667.
[106]Oleksyszyn J,Powers J C,etc.Biochemistry 1991,30:485.
[107]Oleksyszyn J,Boduszek B,Kam C,Powers J C.J.Med.Chem.1994,37:226.
[108]Abuelyaman A S,Jackson D S,Hudig D,etc.Arch.Biochem.Biophys.1997,344(2):271.
[109]Hamilton R,Walker B,Walker B J.Syn Bioorg.Med.Chem.Lett.1998,8:1655.
[2]Fitzpatrick F A,Wheeler R.Int.Immunopharmacol,2003,3:1699.
[3]Newraan D J,Cragg G M,Snader K M.J.Nar.Prod,2003,55:1022.
[4]刘成梅,游海.天然产物有效成分的分离与应用.化学工业出版社.2003.
[5]沈萍.微生物学.北京:高等教育出版社.2000.
[6]黄秀梨.微生物学.北京:高等教育出版社.2003.
[7]周德庆.微生物学教程.北京:高等教育出版社.2002.
[8]Account to:Editors,Antifungal Antibiotics[J].J Antibiot,1996-2005,49-58:1.
[9]张致平.中国处方药,2006,48(3):28.
[10]Sanflard D.Curr Opin Microbiol,2002,5(4):379.
[11]李端等.药理学(第四版).人民卫生出版社.2000.
[12]杨宝峰.药理学(五年制教材).北京医科大学出版社.2003.
[13]李全,谢毓敏.药学进展,1996,20(3):129.
[14]Tremont.Lukats.W,Gilbert MR.Cancer Control,2003,10(2):125.
[15]方美娟.厦门大学傅士毕业论文.2006.
[16]Atalay G,Candoso F,Awada A,Et al.Ann Oncol,2003,14(9):1346.
[17]Singh S B,Lingham R B.Pharmacol Ther,1994,62(1-2):57.
[18]Osterhage C.Dissertation of Technischen Universitat Carolo-Wilhelmina.2001.
[19]Fleming A Br.J.Exp.Pathol,1929,10:226.
[20]Dreyfuss M,Harri E,Hotmann H,Kobel H,Pache W,Tscherter H.Cyclosporin A and C.Eur.J.Appl.Microbiol.Biotechnol,1976,3:125.
[21]Goodman G A,Goodman L S,Rall TW,Murad F.(eds.) Macmillan.7th ed.New York.1985,1298.
[22]Rehm H J,etc.Industrielle Mikrobiologie.2nd ed.Berlin:Springer-Verlag,1980.
[23]Alberts A W,Chen J,Kuron G,etc.Proc.Natl.Acad.Sci.U.S.A,1980,77(7):3957.
[24]于能江,郭顺星.中国药学杂志.1999,34(9):577.
[25]Romila D C,Gerhard S,Jeffrey J,Valerie B,Feng X D,Guy T C.Diazepinomicin.J Nat Prod,2004,67:1431.
[26]Kerstin Liberra and Ulrike Lindequist.Pharmazie,1995,50(9):583.
[27]Ring-Chung cheng,Mustafa varoglu,Leif abrell,etc,J.Org.chem,1994,59:6344.
[28]Atsushi Numata,Chika Takahashi,Yoshinod Ito,etc.Tetrahedron left,1992,33(12):1621.
[29]Wei H,Itoh T,Kinoshita M,Nakai Y,etc.Tetrahedron,2004,60:6015.
[30]Christine K,Stefan K,Anthony D W,Gabriele M K.J Nat Prod,2004,67:1058.
[31]Cueto M,MacMillan JB,Jensen PR,Fenical W.Phytochemistry,2006,67(16):1826.
[32]Jeffrey T G,Taro A,Akiko A,etc.J Nat Prod,2004,67:362.
[33]Varoglu M,Crews P,etc.J.Nat.Prod,2000,63(1):41.
[34]Lee S M,Li X F,Jiang H L,Cheng JG;etc.Tetrahedron Lett,2003,44(42):7707.
[35]Emsley J,Hall D.The chemistry of Phosphorus.London:Harper & Row,1976.
[36]赵玉芬,赵国辉,麻远.磷与生命科学.清华大学出版社,2005.
[37]Chen R-Y,Mao L-J.Chem.J.Chin.Univ.1995,16:892(in Chinese).
[38]陈何如,徐杰诚.药学学报.1996,31(9):671.
[39]李在国,黄润秋,杨昭,邵瑞链.应用化学,1999,02:94.
[40]Wang Q M,Li Z G,Huang R Q,Cheng J R.Heteroat.Chem.2001,12:68.
[41]Liu X J,Chen R Y,Yang Y Y.Chem.J.Chin.Univ.2002,23:1299(in Chinese).
[42]Kristel Senten,Liesbeth Daniels,Pieter Van der Veken,etc.J.Comb.Chem.2003,5:336.
[43]J Joossens,P Vander Veken,A M,etc.J.Med.Chem.2004,47:2411.
[44]Yuan C Y,Wang G H,etc.Synthesis,1990,3:256.
[45]Shi D Q,Chen R.Y.Chin.J.Appl.Chem.2002,19:780(in Chinese).
[46]Seyferth D,Marmor R S,Hilbert P.J.Org.Chem.1971,36(10):1379.
[47]Shono T,Matsumura Y,Tsubata K.Tetra.Letts.1981,22(34):3249.
[48]Natchev I A,etc.Tetrahedron 1988;44(20):6465.
[49]Kudzin Z H,Kotynsi A.Synthesis,1980,1028.
[50]Asano S,Kitahara T,Ogawa T,et al.Agr Biol.Chem,1973,37:193.
[51]Darrell K,Berlin N K,Roy R T,et al.Synthesis,1968,4494.
[52]Darrell K,Berlin N K,Claunch R T,et al.J.Org.Chem,1968,33:3090.
[53]Green D,Elgendy S,Baban J.A,et al.Tetrahedron Letts,1993,34:6917.
[54]Lejczak B,Kafarski P,Mokokowiecka E.Biochem.J,1987,242:81.
[55]Rachon J,Schollkopf U,Wintel T H.Lieb.Ann.Chem,1981,709.
[56]Chalmers M.E,Kosolapoff G.M.J.Am.Chem.Soc.1953,75:5278.
[57]Tyka R,etc.Tetra.Letts.1970,9:677.
[58]Oleksyszyn J,Gruszecka E,etc.Tetra Letts.1981,22(36):3537.
[59]Oleksyszyn J,Tyka R,etc.Tetra.Letts.1977,32:2823.
[60]Kudzin Z H,Stec W J,etc.Synthesis,1978,6:469.
[61]Gajda T,Matusiak M,etc.Syn.Commun,1992,22:2193.
[62]Lukszo J,Tyka R,etc.Synthesis,1977,239.
[63]Green D,Patel G,Elgendy S,et al.Tetrahedron,1994,50:5099.
[64]Gilmore W F,Mcbride HA.J.Am.Chem.Soc.1972,94:4361.
[65]Amos B.Smith,Kraig M.Yager,and Carol M.Taylor.J.Am.Chem.Soc.1995,117:10879.
[66]宁永成.有机化合物结构鉴定与有机波谱学(第二版),北京:科学出版社,2000.
[67]唐恢同.有机化合物光谱鉴定,北京大学出版社,1994.
[68]Burlingame A L,Boyd R K,Gaskell S.T,Anal,Chem,1996,68:599R.
[69]Matsuo T,Seyame Y,J.Mass.Spectrom,2000,35:114.
[70]Wigger Mafia,Eyler John R,etc.J.Am.Soc.Mass Spectrom,2002,13:1162.
[71]Davis G D,etc.Jr.ACC.Chem.Res,1979,12:359.
[72]Munson B,Field F H,etc.J.Am.Chem.Soc,1966,88:2621.
[73]Barber M,Bordoli R S,Scdgwick R D.et al,Nature,1981,293:270.
[74]Fenn J B,Mann M,Meng C K,et al,Science,1989,246:64.
[75]Smith R D,Loo J A,et al,Anal Chem,1990,62:882.
[76]ReidN W,etc.Adv.Mass Spectrom,1980,8:1480.
[77]Karas M,Hillenkamp F,etc.Anal,Chem,1988,60:2299-2301.
[78]Beavis R C,Chait B T,etc.Rapid Commun.Mass Spectrom,1989,3:233.
[79]Hillenkamp F,Karas M,Beavis R C,Anal Chem,1991,63:1193A.
[80]Fenselau C,etc.Anal Chem,1997,69:661A.
[81]Biemann K,etc.Annu.Rev.Biochem,1992,61:977.
[82]Beavis R.C.et al,Anal,Chem,1990,62:1836.
[83]McLafferty F W,etc.J.Am.Soc.Mass Spectrom,1997,8:380.
[84]Sheldrick G M.(1997).SHELXS-97 and SHELXL97.University of Gottingen,Germany.
[85]Accelrys(2001).ViewerPro(V4.2).Accelrys Inc,Burlington,Massachusetts,USA.
[86]Bruker(2001).SAINT(V6.22),SMART(V5.625) and SADABS(V2.03).Bruker AXS Inc,Madison,Wisconsin,USA.
[87]Farrugia L J,etc.J.Appl.Cryst.1997,30:565.
[88]AN Starratt.Phytochem,EN,1976,15:2002.
[89]C.Delseth,Helv.Chim.Acta,EN,1979,62:2037.
[90]Lan D Entwistlf,Colin C Howard,etc.Phytochemstry 1974,13:173.
[91]Singleon V L,Bohonos N,Ullstrup A J,Decumbin.Nature,1958,181:1072.
[92]Betina V,Nemec P,Dobias J,Barath A,Cyanein.Folia Microbiol.1962,7:353.
[93]Suzuki Y,Tanaka H,Aoki H,Tamura T.Agric.Biol.Chem.1979,34:395.
[94]Zhu J W,Hori H,Tsukuda T,etc.Bioorg.Med.Chem.Lett.1997,7:139.
[95]Ahmaol,et al.[J].PYTCAS;Phytochemistry 1976,15:2000.
[96]Delseth,C.et al.HCACAV,Helv.Chim.Acta;EN.1979,62:2037.
[97]Schmidt Bernd;Nave Atefan.Advanced Synthesis & Catalysis.2007,349(1+2):215.
[98]Takahashi H,Yoshioka M,Imai N.Synthesis.1994,6:763.
[99]Takahashi H,Yoshioka M,Imai N.Synthesis 1994,6:763.
[100]Takahiko Akiyama,MachiKo Sanada,Kohei Fuchibe.Synlett 2003,10:1463.
[101]Babak Kaboudin,Rahman Nazari,etc.Tetrahedron Letters.2001,42:8211.
[102]Babak Kaboudin and Khavar Moradia.Tetrahedron Letters.2005,46(17):2989.
[103]方华.厦门大学博士毕业论文.2007.
[104]Hamilton R,Walker B,etc.Tetrahedron Lett.1993,34(17):2847.
[105]Wang C J,Taylor T L,Mical A J,etc.Tetrahedron Lett.1992,33(50):7667.
[106]Oleksyszyn J,Powers J C,etc.Biochemistry 1991,30:485.
[107]Oleksyszyn J,Boduszek B,Kam C,Powers J C.J.Med.Chem.1994,37:226.
[108]Abuelyaman A S,Jackson D S,Hudig D,etc.Arch.Biochem.Biophys.1997,344(2):271.
[109]Hamilton R,Walker B,Walker B J.Syn Bioorg.Med.Chem.Lett.1998,8:1655.