长春碱的提取、分离纯化和半合成研究
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摘要
长春花(Catharanthus roseus (L.) G. Don)是夹竹桃科长春花属一种重要的药用植物。长春花中含有很多有价值的吲哚类生物碱,其中长春碱(Vinblastine, VLB)、长春新碱(Vincristine, VCR)是临床广泛应用的抗肿瘤药物,长春质碱(Catharanthine, CTR)和文多林(Vindoline, VDL)是长春碱类药物半合成的重要原料。本文以提高长春花资源利用率为目的,开展了长春碱类生物碱的提取,分离和半合成研究,建立了长春花综合利用的一个新模式。
1、建立了一个适合工业生产的长春花提取方法,在提取同时得到长春碱,长春质碱和文多林。较佳提取提取工艺为四倍80%乙醇提取,提取三次。
2、通过pH梯度萃取将长春总碱分为双吲哚碱和单吲哚碱两部分,并通过层析分离纯化得到长春碱,长春质碱和文多林。
3、通过重铬酸与长春碱反应一步生成长春新碱,并通过层析分离得到99%长春新碱。
4、通过长春质碱和文多林耦合生成脱水长春碱,并通过溴化、缩环反应得到长春瑞滨,再通过一系列层析分离得到99%的长春瑞滨。
Catharanthus roseus (L.) G. Don is one of the important medicinal plants of Catharanthus in Apocynaceae.Many important indole alkaloids were found in Catha-ranthus roseus,in which Vinblastine(VLB) and Vincristine(VCR) were widely used for anti-cancers in clinic,and Catharanthine(CTR) and Vindoline(VDL) were important materials for semi-synthesis of Catharanthus alkaloids.In this article,In order to im-provethe utilization of Catharanthus roseus's.the process of extraction,isolation and semi-synthesis of VLB were studied,and a mode for comprehensive utilization of Ca-tharanthus roseus was established.
1. Establish a extraction method for Catharanthus roseus in industry,to obtain VLB Catharanthine(CTR) and Vindoline(VDL).The extraction method use four times 80% ethanol-water(v/v) and extraction three times.
2. Vinblastine were separated into mono-indole alkaloids and dimeric-indole al-kaloids by pH gradient extraction method, and VLB Catharanthine(CTR) and Vindo-line(VDL) were obtained by chromatography method.
3. Synthesize Dichromic acid with VLB to obtain Vincristine(VCR) by one-step reaction, and the 99% purity of Vincristine(VCR) was obtained by chromatography method..
4. Couple Catharanthine(CTR) with Vindoline(VDL)'s to obtain Anhydrovinblas-tine(AVLB), then go through bromination and Ring-Closure Reaction-, further go through chromatography separation, then 99% purity of Vinorelbine(VNR) was ob-tained.
1、建立了一个适合工业生产的长春花提取方法,在提取同时得到长春碱,长春质碱和文多林。较佳提取提取工艺为四倍80%乙醇提取,提取三次。
2、通过pH梯度萃取将长春总碱分为双吲哚碱和单吲哚碱两部分,并通过层析分离纯化得到长春碱,长春质碱和文多林。
3、通过重铬酸与长春碱反应一步生成长春新碱,并通过层析分离得到99%长春新碱。
4、通过长春质碱和文多林耦合生成脱水长春碱,并通过溴化、缩环反应得到长春瑞滨,再通过一系列层析分离得到99%的长春瑞滨。
Catharanthus roseus (L.) G. Don is one of the important medicinal plants of Catharanthus in Apocynaceae.Many important indole alkaloids were found in Catha-ranthus roseus,in which Vinblastine(VLB) and Vincristine(VCR) were widely used for anti-cancers in clinic,and Catharanthine(CTR) and Vindoline(VDL) were important materials for semi-synthesis of Catharanthus alkaloids.In this article,In order to im-provethe utilization of Catharanthus roseus's.the process of extraction,isolation and semi-synthesis of VLB were studied,and a mode for comprehensive utilization of Ca-tharanthus roseus was established.
1. Establish a extraction method for Catharanthus roseus in industry,to obtain VLB Catharanthine(CTR) and Vindoline(VDL).The extraction method use four times 80% ethanol-water(v/v) and extraction three times.
2. Vinblastine were separated into mono-indole alkaloids and dimeric-indole al-kaloids by pH gradient extraction method, and VLB Catharanthine(CTR) and Vindo-line(VDL) were obtained by chromatography method.
3. Synthesize Dichromic acid with VLB to obtain Vincristine(VCR) by one-step reaction, and the 99% purity of Vincristine(VCR) was obtained by chromatography method..
4. Couple Catharanthine(CTR) with Vindoline(VDL)'s to obtain Anhydrovinblas-tine(AVLB), then go through bromination and Ring-Closure Reaction-, further go through chromatography separation, then 99% purity of Vinorelbine(VNR) was ob-tained.
引文
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[3]国家中医药管理局.中华本草[M].上海：上海科技出版社,1998,6:281.
[4]Noble RL,Beer CT,Cutts JH. Further biological activities of vincaleukoblastine: an alkaloid isolated from Vinca rosea(L.)[J].Biochemical Pharmacology,1959,1(4): 347-348.
[5]Svoboda G.H. Alkaloids of Vica rosea Linn.Ⅸ:Extraction and characterization of leurosidine and leucocristine[J]. Lloydia,1961,24:173-178.
[6]祖元刚,罗猛.长春花生物碱成分及其药理作用研究进展[J].天然产物研究与开发,2006,18(2):325-329,294.
[7]Jacquesy J.C.,Fahy J..Biomedical chemistry:applying chemical principles to und-erstanding and treatment of disease [M]. New York:Joun Wiley and Sons Inc,2000.
[8]Donehower R.C.,Rowinsky E.K..Anticancer drugs derived from plants[M].Phila-delphia:J.B.Lippincott Company,1993.
[9]Johnson S.I.,Amstrong J.G.,Gorman M.,et al..The Vinca alkaloids:a new class of oncolytic agents[J].Cancer Research,1996,23(81):1390-1427.
[10]王非,李雷鸿,孙家宝等.人为采收后长春花中长春碱含量的动态变化[J].东北林业大学学报,2005,33(4)：42-44.
[11]Morman R,Farnsworth P.The pharmacognosy of the periwinkles,Vinca and Catharanthus[J].Lloydia,1961,24:105-138.
[12]卢懿,侯世祥,陈彤.长春花抗癌成分长春新碱研究的进展[J].中国中药杂志,2003,28(11)：1006-1008.
[13]Endo T. Good body, Vikovic J., et al. Enzymes from catharanthus Rosens Cell cultures That Couplevindoline and Cathranthine to from 3',4'-anhydrovinblatine[J]. Phytochem,1998,49(27):2147.
[14]Kutney J.P.,Boulet G.A.,et al.Alkaloid Production in Catharanthus roseus (L.) G. don Cell Cultures.VI.Biotransformation of 3,4-anhydrovinblastine with Catharanthus roseus Cell cultures and Enzyme Systems[J]. Heterocycles,1998,47(27):629.
[15]Serap whitmer, Camilo Camel, Didier Hallard,et al..Influence of Precursor A vai-lability on Alkaloid Accumulation by Transgenic Cell Line of Cathranthus Rosus[J]. Plant Physiol,1998,116(2):853.
[16]Nadine Imbault, Martine Thiersault,Paulette Duperon,et al..Pravastatin:A Tool for Investigating the Availability of Mevelonate Metabolitie for Primary and Secon-dary Metabolism in Catharanthus roseus Cell Suspensions[J].Physiologia Plantarum, 1996,98(4):803.
[17]Rajiv Bhadra, John A. Morgan, Jacqueline V shanks.Transient Studies of Light-adapted Cultures of Hairy Roots of Catharanthus roseus:Growth and Indole Al-kaloid Accumulation[J]. Biotechnol and Bioeng,1998.60(6):670.
[18]Fahy J.,Duflos A.,Ribet J.P.,et al..Vica alkaloids in superacdic media:a method for creating a new family of antitumor derivatives[J] Journal of Amercian Chemical Society,1977,119(36):8576-8577.
[19]Lobert S.,Vulevic B.,Correia J.J..Interaction of Vica alkaloids with tubulin-a conparison of vinblastine,vincristine and vinorelbine[J].Biochemistry,1996,35(21): 6806-6814.
[20]Jordan M.A., Wilson L..Kinetic analysis of tubulin exchange at microtubule ends at low vinblastine concentrations[J].Biochemistry,1990,29(11):2730-2739.
[21]Na G.C.,Timasheff S.N..In vitro vinblastine-induced tubulin paracrystals[J].Journal of Molecular Biology.1982,187(1):61-73.
[22]Tellingen V.O.,Sips J.M.,Beijnen J.H.,et al..Pharmacology,bio-analysis and Phar-ma-cokinetics of the Vinca alkaloids and sem-synthetic deritatives[J].Anticancer Re-search,1992,12:1699-1716.
[23]Zhou X.J.,Rahmani R..Preclinical and clinical pharmacology of Vica alkaloids[J]. Drug,1992,44(1):1-16.
[24]贾彩云,纪人明.抗肿瘤新药—艾西克在临床应用中的毒副反应[J].实用肿瘤学杂志,1994,8(4):56-60.
[25]Zhang H.Z.,Dong Z.H.,She J.,et al..Modern study of traditional Chinese medicine Vol Ⅱ [M].Beijing:Academic Press,1991:1020-1035.
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