东北红豆杉枝叶的化学成分研究
【网学提醒】：本文主要为网上学习者提供东北红豆杉枝叶的化学成分研究，希望对需要东北红豆杉枝叶的化学成分研究网友有所帮助，学习一下吧!
资料包括： 论文(28页9670字)
说明：
摘要：抗癌药物紫杉醇的发现是近20年来天然药物化学研究领域中的划时代事件，研究涉及的学科、学者之多都是空前的。紫杉醇具有独特的抗癌机理，临床应用广泛，疗效好，而红豆杉中紫杉醇含量很低，不能满足紫杉醇等活性成分的临床需要量。
为了寻找其它可供利用的资源和有效成分，多年来全世界学者对红豆杉属植物的化学成分进行了深入的研究，获得了一系列结构新颖的化合物。我国东北红豆杉资源丰富，为了寻找紫杉醇和其它有效成分，同时又不破坏资源，我们对东北红豆杉枝叶的化学成分进行了研究。
本论文利用开放硅胶柱色谱和凝胶柱色谱及重结晶等技术对东北红豆杉枝叶75%乙醇冷浸提取物进行分离，分离得到17个化合物，根据化合物的理化性质和光谱数据确定了15个化合物的结构，分别为紫杉双黄酮 A (1)、穗花杉双黄酮 (2)、槲皮素 (3)、山柰酚 (4)、紫杉叶素 (5)、二氢山柰酚 (6)、紫杉宁 (7)、紫杉宁A (8)、紫杉宁B (9)、taxacin (10)、坡那甾酮A (11)、胡萝卜苷 (12)、红豆杉苷 (13)、对羟基苯甲醛 (14)、对羟基苯甲酸 (15)。其中化合物紫杉双黄酮 A (1)为一未见文献报道的新化合物，化合物紫杉叶素 (5)、二氢山柰酚 (6)、对羟基苯甲醛 (14)、对羟基苯甲酸 (15)为首次从该植物中分离得到，化合物taxacin (10)利用二维核磁共振技术首次对其氢碳数据进行了全归属。
关键词：红豆杉，东北红豆杉枝叶，紫杉双黄酮 A，化学成分
ABSTRACT：The discovery of paclitaxel with its remarkable anti-cancer activity has been a distinguished progress in natural product chemistry during the past 20 years. Paclitaxel is a widely used medicine due to its excellent effects; however, it couldn’t supply clinical application as a result of its low yield in Taxus plants. Twigs and leaves are considered to be the promising sources of taxane-type diterpenoids because they can be collected without the destruction of the trees. We have further investigated the chemical constituents of the twigs and leaves from Taxus cuspidata which grows in Liaoning province.
This paper described the isolation and structure elucidation of the constituents from the twi-
gs and leaves of Taxus cuspidata Sieb.et Zucc. The compounds were isolated by chromatography
on silica gel and Sephadex LH-20 columns and NMR spectral analysis were employed for their structure identification. 17 compounds were obtained and 15 of them were elucidated as: taxusfl-
avone A (1), amentoflavone (2), quercetin (3), kaempferol (4), taxifolin (5), dihydrokaempferol (6), taxinine (7), taxinine A (8), taxinine B (9), taxacin (10), ponasterone A (11), daucosterol (12), taxicatin (13), p-hydroxy benzaldehyde (14), p-hydroxybenzcic acid (15), respectively. Among them, compound 1 was a novel compound, and compound 5, 6, 14, 15 were isolated from this plant for the first time, furthermore, the proton and carbon signals of taxacin were assigned by means of 2D NMR techniques including HMBC and HMQC for the first time, too.
Keywords: Taxus; Taxus cuspidata Sieb.et Zucc; taxusflavone A; chemical constituents;
第一章 东北红豆杉的化学成分与药理活性研究进展
第一节 红豆杉的地理分布及中医药研究
抗癌药物紫杉醇的发现是近20年来天然药物化学研究领域中的划时代事件，研究涉及的学科、学者之多都是空前的。紫杉醇主要是从红豆杉的树皮中来提取。具有独特的抗癌机理，临床应用广泛，疗效好。现对红豆杉属植物的地理分布及中医药情况作如下综述。
1地理分布情况
全世界的红豆杉属植物根据其生长地域和生物学特性可分为11个种类，除澳洲的Austrotaxus spicata产于南半球外，其余分别分布于北半球的温带至亚热带地区。我国Taxus属植物资源丰富，有西藏红豆杉(T. wallichiann Zucc )、云南红豆杉(T. yunnanensis Cheng et L. K. Fu )、南方红豆杉(T. chinensis var. mairei cheng et L. K. Fu，又称美丽红豆杉)、东北红豆杉(T. cuspidata Sieb et Zucc，又称日本紫杉Japanese yew）和红豆杉(T. chinensis (Pilger) Rh.d )，共4种和1个变种。东北红豆杉（Taxus）又名紫杉，赤柏松，为紫杉科紫杉属长绿针叶乔木或灌木，是第四纪冰川遗留下来的古老树种，在地球上已有250万年的历史。其主要分布于我国的辽宁省宽甸、凤城、本溪地区；吉林省安图、抚松、白山、长白、集安地区；黑龙江省仅见于铁力县伊吉密河及穆棱一带东部山区。此外，在亚洲的日本、朝鲜等也有分布。
目录：
中文摘要 …………………………………………………………………………1
英文摘要 …………………………………………………………………………2
第一章 东北红豆杉的化学成分与药理活性研究进展 … ……………………………3
第一节 红豆杉的地理分布及中医药研究……………………………………………3
第二节 东北红豆杉的化学成分研究…………………………………………………3
第三节 药理活性研究进展 ………………………………………………………16
第二章 东北红豆杉枝叶的化学成分研究 …………………………………………18
第一节 概述 ……………………………………………………………………18
第二节 化合物的结构解析 ………………………………………………………20
第三章 实验部分 ………………………………………………………………38
第四章 结果与讨论 ……………………………………………………………45
参考文献 … ………………………………………………………………………46
致谢 ……………………………………………………………………………51
附图 ……………………………………………………………………………52
参考文献：
江苏新医学院。《中药大辞典》。下册，上海人民出版社，1997: 2342.
黄泰康，丁志遵，赵守训等。《现代本草纲目》。上卷，中国中医药科技出版社，2000: 1176-1177.
中国药科大学。《中药辞海》。第一卷，中国医药科技出版社，1993: 2395-2396.
《全国中草药汇编》编写组。全国中草药汇编。上册，人民卫生出版社，1999: 473-474.
肖培根，李大鹏，杨世林。《新编中药志》.第三卷，化学工业出版社，2002: 719-734.
Wani M.C, Taylor H L, Wall M. E, Coggon P, McPhail, A T. The isolation and structure elucidation of taxol, a novel anti-leukemic and anti-tumor agent from Taxus brevifolia. J. Am. Chem. Soc. 1971, 93: 2325 - 2327.
Shi QW. Isolation and structural elucidation of taxoids from the Japanese and the Chinese yews. Doctoral thesis, Tohoku University, 2000, Sendai, Japan.
Chiang H.C, Woods M.C, Nakadaira Y, et al. The structures of four new taxinine congeners and a photo-
chemical transannular-reaction. Chem Commun, 1967；1201-1202.
佟晓杰，方唯硕，周金云等。东北红豆杉枝叶化学成分研究。药学学报, 1994, 29: 55- 60.
Baloglu E, Kingston DGI. The taxane diterpenoids. Nat Prod, 1999, 62: 1448-1472.
Kosugi K, Sakai J, Zhang S, et al . Neutral taxoids from Taxus cuspidata as modulators of multidrug-
resistant tumor cells. Phytochemistry, 2000, 54: 839-845.
Kingston DGI, Molinero AA, Rimoldi J M. In: Herz W, Moore RE, Kirby GW, et al. Progress in the Chemistry of Organic Natural Products. Springer: Wien, 1993, 61: 1-206.
Miyazaki M,Shimizu K,Mishima H, et al. The constituents of the seeds of Japan Yew (Taxus cuspidata)
Chem Pharm Bull, 1968, 16: 546-548.
Kobayashi J, Inubushi A , Hosoyama H, et al. Taxuspine E-H and J new taxoids from Japanese yew (Taxus cuspidata) Tetrahedron, 1995, 51: 5971-5978.
[15] Shigemori H, Wang XX, Yoshida N, et al. Taxuspine X-Z new taxoids from Japanese yew (Taxus cuspidata). Chem Pharm Bull, 1997, 45: 1205-1208.
[16] Morita H, Gonda A, Wei L, et al . Four new taxoids from Taxus cuspidata var. nana. Planta Med, 1998, 64: 183-186.
[17] Morita H, Gonda A, Wei L, et al . Taxoids from Taxus cuspidata var. nana. Phytochemistry 1998, 48:
857-862.
[18] Wang XX, Shigemori H, Kobayashi J . Taxezopidines B-H, newtaxoids from Japanese yew (Taxus cuspidata). J Nat Prod, 1998, 61: 474-479.
[19] Tong XJ , Fang WS , Zhou JY, et al. Three new taxane diterpenoids from needles and stems of Taxus cuspidata. J Nat Prod, 1995, 58: 233-238.
[20] Yeh MK,Wang JS, Liu LP, et al . A new taxane derivative from the heartwood of Taxus mairei. Phyto-
chemistry, 1988, 27: 1534-1536.
[21] Della Casa de Marcano DP, Halsall TG. The isolation of seven new taxane derivatives from the heartwood of yew (Taxus baccata L) Chem Commun, 1969: 1282-1283.
[22] Liang JY, Min ZD, Niwa MStudies on the diterpenes of Taxus mairei Ⅱ. Structure of2-deacetoxytaxi-
nine J Acta Chim Sin,1988, 46: 1053-1054.
[23] Kobayashi J, Shigemori H. Bioactive taxoids from the Japanese yew (Taxus cuspidata). Med Res Rev, 2002, 22: 305-328.
[24] Barboni L, Gariboldi P, Appendino G, et al . New taxoids fromTaxus baccata. Liebigs Ann, 1995, (3):
345-349.
[25] Sugiyama T, Oritani T,Oritani T. A novel taxane diterpenoid from the needles of Japan yew, (Taxus cuspidata). Biosci Biotech Biochem, 1994, 58: 1923-1924.
[26] Parmar VS, J ha A. Chemical Constituents of Taxus Species. In: AttaurRahman (Ed), Studies in Natural Product Chemistry, Vol 20. Elsevier Science B. V, 1998: 79-133.
[27] Shi QW,Petake TL, Sauriol F, et al . Taxanes in rooted cuttings vs. mature Japanese yew. Can J Chem, 2003, 90: 64-74.
[28] Wang XX, Shigemori H, Kobayashi J. Taxuspines Q, R, S and T, new taxoids from Japanese yew (Taxus cuspidata). Tetrahedron, 1996, 52: 12159-12164.
[29] Shigemori H, Sakurai CA, Hosoyama H, et al . Taxezopidines J, K and L, new taxoids from Taxus cuspidata inhibiting Ca2+-induced depolymerization of microtubules. Tetrahedron, 1999, 55: 2553-2558.
[30] Yoshizaki F, Fukuda M, Hisamichi S, et al . Stucture of taxane diterpenoids from the seeds of Japan yew (Taxus cuspidata). Chem Pharm Bull, 1988, 36: 2098-2102.
[31] Nizam UK,Nazneen P. The constituents of genus Taxus. Journal of Scientific & Industrial Research
1987, 46: 512-516.
[32] Beutler JA, Chmurny GM,Look SA, et al. Taxinine M, a new tetracyclic taxane from Taxus brevifolia. J Nat Prod, 1991, 54: 893-897.
[33] Barboni L, Gariboldi P, Torregiani E, et al. Minor taxoids from Taxus wallichiana. J Nat Prod, 1995, 58: 934-939.
[34] Hosoyama H, Inubushi A, Katsui T. Taxuspines U, V and W, new taxanes and related diterpenoids from Japanese yew (Taxus cuspidata). Tetrahedron, 1996, 52: 13145-13150.
[35] Shi QW,Oritani T,Horiguchi T, et al . Four novel taxane diterpenoids from the needles of Japanese yew (Taxus cuspidata).Biosci Biotechnol Biochem, 1999, 63: 924-929.
[36] Wang XX,Shigemori H , Kobayashi J . Taxezopidine A, a novel taxoid from Japanese yew, (Taxus cuspidata). Tetrahedron Lett, 1997, 38: 7587-7588.
[37] Morita Gonda A. Wei LYamamura Y. Takeya Itok Awa. Journal of Natural Products 1997, 46, 583.
[38] Shi QW, Oritani T, Kiyota H, et al. Taxane diterpenoids fromTaxus yunnanensis and Taxus cuspidata. Phytochemistry, 2000, 54: 829-834.
[39] Shi QW, Cao CM, Gu JS , et al. Four new taxane metabolites from the needles of Taxus cuspidata. Nat Prod Res, 2006, 20: 173-179.
[40] Kosugi K, Sakai J, Zhang S, et al. Neutral taxoids from Taxuscuspidata as modulators of multidrug
-resistant tumor cells. Phytochemistry, 2000, 54: 839-845.
[41] Kobayashi J, Hosoyama H, Shigemori H, et al. Taxuspine D, a new taxane diterpene from Taxus cuspidata with potent in hibitory activity against Ca2 + induced depolymerization of microtubule.
Experientia, 1995, 51: 592-595.
[42] Kobayashi J, Ogiwara A, Hosoyama H, et al . Taxuspine A-C, new taxoids from Japan yew Taxus cuspidata inhibiting drug transport activicity of P-glycorprotein in multidrug-resistant cells. Tetrahedron, 1994, 50: 7401-7416.
[43] Wang XX, Shigemori H, Kobayashi J. Taxuspine K, L and M,new taxoids from Japanese yew (Taxus cuspidata). Tetrahedron, 1996, 52: 2337-2342.
[44] Kobayashi J, Hosoyama H, Katsui T, et al. Taxuspine N, O and P new taxoids from Japanese yew Taxus cuspidata. Tetrahedron. 1996, 52: 5391-5396.
[45] Appendino G, Borboni L, Gariboldi P, et al . Revised structure of brevifoliol and some Baccatin Ⅵ derivatives. J Chem Soc, Chem Commun, 1993: 1587-1589.
[46] Barboni L, Gariboldi P, Torregiani EGA, et al. Taxanes from the needles of Taxus wallichiana. Phytochemistry, 1993, 33: 145-150.
[47] Murakami R, Shi QW,Horiguchi , et al. A novel rearranged taxoid from needles of Japanese yew (Taxus cuspidata). BiosciBiotechnol Biochem1999, 63: 1660-1663.
[48] Morita H, Gonda A, Wei L, et al. Taxuspinananes A and B, new taxoids from Taxus cuspidata var nana. J Nat Prod, 1997, 60: 390-392.
[49] Fuji K, Tanaka K, Li B, et al. Novel diterpenoids from Taxus chinensis. J Nat Prod, 1993, 56: 1520-1531.
[50] Rao K V, Johnson J H.occurrence of 2, 6-dimethoxy cinnamaldehyde in Taxus floridana and structural revision of taxiflorine to taxchinin M. Phytochemistry, 1998, 49: 1361-1364.
[51] Morita H, Gonda A, Wei L, et al . Four new taxoids from Taxus cuspidata var. nana. Planta Med, 1998, 64: 183-186.
[52] Barboni L, Gariboldi P, Appendino G, et al. New taxoids fromTaxus baccata. Liebigs Ann, 1995, (3):
345-349.
[53] Shi QW, Oritani T,Hiromasa K, et al . Three new taxoids fromthe leaves of the Japanese yew, Taxus
cuspidata. Nat Prod Lett, 2001, 15: 55-62.
[54] Morita H, Gonda A, Wei L, et al . Taxoids from Taxus cuspidate var. nana. Phytochemistry 1998,
48: 857-862.
[55] Shi QW, Li ZP, Zhao D, et al. New 2 (3 -20) abeotaxanes and 3, 11-cyclotaxane from needles of Taxus cuspidata. BiosciBiotechnol Biochem, 2004, 68: 1584-1587.
[56] Sakai J, Sasaki H, Kosugi K, et al. Two taxoids Taxus cuspidata as modulartors of multidrug resistant tumor cells. Heterocycles, 2001, 54: 999-1009.
[57] Shi QW,Oritani T,Sugiyama T, et al. Three new bicyclic taxane diterpenoids from the needles of Japanese yew ,(Taxus cuspidata). J Asian Nat Prod Res, 1999, 2: 63-70.
[58] Murakami R, Shi QW,Oritani T. A taxoid from the needles of the Japanese yew, Taxus cuspidata. Phytochemistry, 1999, 52: 1577-1580.
[59] Nguyen TD, Banskota AH, Tezuka Y, et al. Diterpenes and sesquiterpenes from the bark of Taxus yunnanensis. Phytochemistry, 2003, 64: 1141-1147.
[60] Ripa PV, Matin EA,Cocciolone SM, et al. Fluctuation of phytoecdysteroids in developing shoots of Tasxus cuspidata. Phytochemistry, 1990, 29 :425-427.
[61] Parmer VS, Jha A ,Bisht KS, et al . Constituents of the yew trees. Phytochemistry 1999, 50: 1267-
1304.
[62] Yoshizaki F, Fufuda M, Hisamichi S. Principal constituents of the seeds of Japan yew (Taxus cuspidata). Shoyakugaku Zashi, 1986, 40: 429.
[63] Erdtman H, Tsuno K. Taxus heartwood constituents. Phytochemistry, 1969, 8: 931-932.
[64] Shi QW taxanes in rooted cutting vs mature Japanese yew Canadian Journal of chemistry 2003. 81(4)
338
[65] 张宏桂，张宏，阎吉昌等。东北红豆杉枝叶化学成分研究 人参研究, 1995, 4.
[66] Kobayashi J, Hosoyama H,Wang XX,et al. Modulation of multi-drug resistance by taxuspine C and other taxoids from Japanese yew. Bioorg Med Chem Lett, 1997, 7: 393-398.
[67] Masahiko Oshige, a, Mika Takenouchi,d Yasutaro Kato,Bioorganic & Medicinal Chemistry 12 (2004) 2597–2601.
[68] 刘海青, 林瑞超, 马双成等。旱生卷柏化学成分的研究 (Ⅰ)。 中草药，2003：34（4）298-299。
[69] 许婧, 李铣, 张鹏等。金刚藤的黄酮类化学成分。沈阳药科大学学报。2004：21（6）424-425。
[70] 穆丽华, 张东明。紫荆化学成分的研究。中国中药杂志2006，3 (21)：1795-1797。
[71] 丁林生，梁侨丽，腾艳芬。枳棋子黄酮类成分研究。药学学报1997，32 (8)：600-602。
[72] 周金云，方起程。天然紫杉烷类二萜类化合物的核磁共振氢谱规律。植物学报1997，39 (5)：467-76。
[73] Yaeko konda .tomomistu Sasaki Absolute structure of 2′ desacetoxyaustrospicatine established by x-ray analysis and circular dichroism spectroscopy . Chem Pharm Bull, 1994: 2621-2623
[74] 霍长虹，李作平，史清文等。天然紫杉烷类二萜类化合物的核磁共振氢谱特征。波谱学杂志 2007，24 (1)：101-118。
[75] 梅文莉, 曾艳波, 丁中涛等。红树植物卤蕨化学成分的分离与鉴定。中国药物化学杂志。2006： 46-48。
[76] 张君增，方起程，梁哓天。中国特有植物白豆杉的化学成分研究。植物学报1996，38 (5)：399-340。
[77] 肖海涛, 李铣。半枝莲的化学成分。沈阳药科大学学报 2006，23 (10)：637-639。
[78] 汤海峰 ，易杨华 ，姚新生等。褐藻果叶马尾藻化学成分的研究《中国海洋药物》2002，6：11-15。
作者点评：
1. 近年来，科学家研究发现一些紫杉烷类化合物虽然其本身没有抗肿瘤活性，但可以作为多药耐药逆转剂改变耐药细胞的耐药性。如紫杉宁及其衍生物，此类化合物具有较高的逆转多药耐药活性，并且具有很低的细胞毒性，这对于将其发展成为新型多药耐药逆转剂是很有利的，因紫杉宁在东北红豆杉枝叶中含量较高，可利用其做为先导化合物进行结构修饰，以便寻找具有较高的逆转多药耐药活性的目标化合物。
2 用开放硅胶柱色谱和凝胶柱色谱及重结晶等技术对东北红豆杉枝叶75%乙醇冷浸提取物进行分离，分离得到17个化合物。根据化合物的理化性质和光谱数据确定了15个化合物的结构，分别为紫杉双黄酮 A (1)、穗花杉双黄酮 (2)、槲皮素 (3)、山柰酚 (4)、紫杉叶素 (5)、二氢山柰酚 (6)、紫杉宁 (7)、紫杉宁A (8)、紫杉宁B (9)、taxacin (10)、坡那甾酮A (11)、胡萝卜苷 (12)、红豆杉苷 (13)、对羟基苯甲醛 (14)、对羟基苯甲酸 (15)。其中化合物紫杉双黄酮 A为一未见文献报道的新化合物，化合物紫杉叶素、二氢山柰酚、对羟基苯甲酸、对羟基苯甲醛为首次从该植物中分离得到，化合物taxacin利用二维核磁共振技术首次对其氢碳数据进行了全归属。
3 传统中医药中以红豆杉属植物东北红豆杉T. cuspidata的枝叶入药，中药处方名紫杉、赤柏松、紫柏松。《中药大辞典》记载：红豆杉中含有的生物碱有利尿、通经、降压、降糖、杀虫、止痒作用，对治疗肾脏病、糖尿病、风湿病和白血病等均有良好疗效。也可从中医药的角度对其药效作用的物质基础进行研究。
资料包括： 论文(28页9670字)
说明：
摘要：抗癌药物紫杉醇的发现是近20年来天然药物化学研究领域中的划时代事件，研究涉及的学科、学者之多都是空前的。紫杉醇具有独特的抗癌机理，临床应用广泛，疗效好，而红豆杉中紫杉醇含量很低，不能满足紫杉醇等活性成分的临床需要量。
为了寻找其它可供利用的资源和有效成分，多年来全世界学者对红豆杉属植物的化学成分进行了深入的研究，获得了一系列结构新颖的化合物。我国东北红豆杉资源丰富，为了寻找紫杉醇和其它有效成分，同时又不破坏资源，我们对东北红豆杉枝叶的化学成分进行了研究。
本论文利用开放硅胶柱色谱和凝胶柱色谱及重结晶等技术对东北红豆杉枝叶75%乙醇冷浸提取物进行分离，分离得到17个化合物，根据化合物的理化性质和光谱数据确定了15个化合物的结构，分别为紫杉双黄酮 A (1)、穗花杉双黄酮 (2)、槲皮素 (3)、山柰酚 (4)、紫杉叶素 (5)、二氢山柰酚 (6)、紫杉宁 (7)、紫杉宁A (8)、紫杉宁B (9)、taxacin (10)、坡那甾酮A (11)、胡萝卜苷 (12)、红豆杉苷 (13)、对羟基苯甲醛 (14)、对羟基苯甲酸 (15)。其中化合物紫杉双黄酮 A (1)为一未见文献报道的新化合物，化合物紫杉叶素 (5)、二氢山柰酚 (6)、对羟基苯甲醛 (14)、对羟基苯甲酸 (15)为首次从该植物中分离得到，化合物taxacin (10)利用二维核磁共振技术首次对其氢碳数据进行了全归属。
关键词：红豆杉，东北红豆杉枝叶，紫杉双黄酮 A，化学成分
ABSTRACT：The discovery of paclitaxel with its remarkable anti-cancer activity has been a distinguished progress in natural product chemistry during the past 20 years. Paclitaxel is a widely used medicine due to its excellent effects; however, it couldn’t supply clinical application as a result of its low yield in Taxus plants. Twigs and leaves are considered to be the promising sources of taxane-type diterpenoids because they can be collected without the destruction of the trees. We have further investigated the chemical constituents of the twigs and leaves from Taxus cuspidata which grows in Liaoning province.
This paper described the isolation and structure elucidation of the constituents from the twi-
gs and leaves of Taxus cuspidata Sieb.et Zucc. The compounds were isolated by chromatography
on silica gel and Sephadex LH-20 columns and NMR spectral analysis were employed for their structure identification. 17 compounds were obtained and 15 of them were elucidated as: taxusfl-
avone A (1), amentoflavone (2), quercetin (3), kaempferol (4), taxifolin (5), dihydrokaempferol (6), taxinine (7), taxinine A (8), taxinine B (9), taxacin (10), ponasterone A (11), daucosterol (12), taxicatin (13), p-hydroxy benzaldehyde (14), p-hydroxybenzcic acid (15), respectively. Among them, compound 1 was a novel compound, and compound 5, 6, 14, 15 were isolated from this plant for the first time, furthermore, the proton and carbon signals of taxacin were assigned by means of 2D NMR techniques including HMBC and HMQC for the first time, too.
Keywords: Taxus; Taxus cuspidata Sieb.et Zucc; taxusflavone A; chemical constituents;
第一章 东北红豆杉的化学成分与药理活性研究进展
第一节 红豆杉的地理分布及中医药研究
抗癌药物紫杉醇的发现是近20年来天然药物化学研究领域中的划时代事件，研究涉及的学科、学者之多都是空前的。紫杉醇主要是从红豆杉的树皮中来提取。具有独特的抗癌机理，临床应用广泛，疗效好。现对红豆杉属植物的地理分布及中医药情况作如下综述。
1地理分布情况
全世界的红豆杉属植物根据其生长地域和生物学特性可分为11个种类，除澳洲的Austrotaxus spicata产于南半球外，其余分别分布于北半球的温带至亚热带地区。我国Taxus属植物资源丰富，有西藏红豆杉(T. wallichiann Zucc )、云南红豆杉(T. yunnanensis Cheng et L. K. Fu )、南方红豆杉(T. chinensis var. mairei cheng et L. K. Fu，又称美丽红豆杉)、东北红豆杉(T. cuspidata Sieb et Zucc，又称日本紫杉Japanese yew）和红豆杉(T. chinensis (Pilger) Rh.d )，共4种和1个变种。东北红豆杉（Taxus）又名紫杉，赤柏松，为紫杉科紫杉属长绿针叶乔木或灌木，是第四纪冰川遗留下来的古老树种，在地球上已有250万年的历史。其主要分布于我国的辽宁省宽甸、凤城、本溪地区；吉林省安图、抚松、白山、长白、集安地区；黑龙江省仅见于铁力县伊吉密河及穆棱一带东部山区。此外，在亚洲的日本、朝鲜等也有分布。
目录：
中文摘要 …………………………………………………………………………1
英文摘要 …………………………………………………………………………2
第一章 东北红豆杉的化学成分与药理活性研究进展 … ……………………………3
第一节 红豆杉的地理分布及中医药研究……………………………………………3
第二节 东北红豆杉的化学成分研究…………………………………………………3
第三节 药理活性研究进展 ………………………………………………………16
第二章 东北红豆杉枝叶的化学成分研究 …………………………………………18
第一节 概述 ……………………………………………………………………18
第二节 化合物的结构解析 ………………………………………………………20
第三章 实验部分 ………………………………………………………………38
第四章 结果与讨论 ……………………………………………………………45
参考文献 … ………………………………………………………………………46
致谢 ……………………………………………………………………………51
附图 ……………………………………………………………………………52
参考文献：
江苏新医学院。《中药大辞典》。下册，上海人民出版社，1997: 2342.
黄泰康，丁志遵，赵守训等。《现代本草纲目》。上卷，中国中医药科技出版社，2000: 1176-1177.
中国药科大学。《中药辞海》。第一卷，中国医药科技出版社，1993: 2395-2396.
《全国中草药汇编》编写组。全国中草药汇编。上册，人民卫生出版社，1999: 473-474.
肖培根，李大鹏，杨世林。《新编中药志》.第三卷，化学工业出版社，2002: 719-734.
Wani M.C, Taylor H L, Wall M. E, Coggon P, McPhail, A T. The isolation and structure elucidation of taxol, a novel anti-leukemic and anti-tumor agent from Taxus brevifolia. J. Am. Chem. Soc. 1971, 93: 2325 - 2327.
Shi QW. Isolation and structural elucidation of taxoids from the Japanese and the Chinese yews. Doctoral thesis, Tohoku University, 2000, Sendai, Japan.
Chiang H.C, Woods M.C, Nakadaira Y, et al. The structures of four new taxinine congeners and a photo-
chemical transannular-reaction. Chem Commun, 1967；1201-1202.
佟晓杰，方唯硕，周金云等。东北红豆杉枝叶化学成分研究。药学学报, 1994, 29: 55- 60.
Baloglu E, Kingston DGI. The taxane diterpenoids. Nat Prod, 1999, 62: 1448-1472.
Kosugi K, Sakai J, Zhang S, et al . Neutral taxoids from Taxus cuspidata as modulators of multidrug-
resistant tumor cells. Phytochemistry, 2000, 54: 839-845.
Kingston DGI, Molinero AA, Rimoldi J M. In: Herz W, Moore RE, Kirby GW, et al. Progress in the Chemistry of Organic Natural Products. Springer: Wien, 1993, 61: 1-206.
Miyazaki M,Shimizu K,Mishima H, et al. The constituents of the seeds of Japan Yew (Taxus cuspidata)
Chem Pharm Bull, 1968, 16: 546-548.
Kobayashi J, Inubushi A , Hosoyama H, et al. Taxuspine E-H and J new taxoids from Japanese yew (Taxus cuspidata) Tetrahedron, 1995, 51: 5971-5978.
[15] Shigemori H, Wang XX, Yoshida N, et al. Taxuspine X-Z new taxoids from Japanese yew (Taxus cuspidata). Chem Pharm Bull, 1997, 45: 1205-1208.
[16] Morita H, Gonda A, Wei L, et al . Four new taxoids from Taxus cuspidata var. nana. Planta Med, 1998, 64: 183-186.
[17] Morita H, Gonda A, Wei L, et al . Taxoids from Taxus cuspidata var. nana. Phytochemistry 1998, 48:
857-862.
[18] Wang XX, Shigemori H, Kobayashi J . Taxezopidines B-H, newtaxoids from Japanese yew (Taxus cuspidata). J Nat Prod, 1998, 61: 474-479.
[19] Tong XJ , Fang WS , Zhou JY, et al. Three new taxane diterpenoids from needles and stems of Taxus cuspidata. J Nat Prod, 1995, 58: 233-238.
[20] Yeh MK,Wang JS, Liu LP, et al . A new taxane derivative from the heartwood of Taxus mairei. Phyto-
chemistry, 1988, 27: 1534-1536.
[21] Della Casa de Marcano DP, Halsall TG. The isolation of seven new taxane derivatives from the heartwood of yew (Taxus baccata L) Chem Commun, 1969: 1282-1283.
[22] Liang JY, Min ZD, Niwa MStudies on the diterpenes of Taxus mairei Ⅱ. Structure of2-deacetoxytaxi-
nine J Acta Chim Sin,1988, 46: 1053-1054.
[23] Kobayashi J, Shigemori H. Bioactive taxoids from the Japanese yew (Taxus cuspidata). Med Res Rev, 2002, 22: 305-328.
[24] Barboni L, Gariboldi P, Appendino G, et al . New taxoids fromTaxus baccata. Liebigs Ann, 1995, (3):
345-349.
[25] Sugiyama T, Oritani T,Oritani T. A novel taxane diterpenoid from the needles of Japan yew, (Taxus cuspidata). Biosci Biotech Biochem, 1994, 58: 1923-1924.
[26] Parmar VS, J ha A. Chemical Constituents of Taxus Species. In: AttaurRahman (Ed), Studies in Natural Product Chemistry, Vol 20. Elsevier Science B. V, 1998: 79-133.
[27] Shi QW,Petake TL, Sauriol F, et al . Taxanes in rooted cuttings vs. mature Japanese yew. Can J Chem, 2003, 90: 64-74.
[28] Wang XX, Shigemori H, Kobayashi J. Taxuspines Q, R, S and T, new taxoids from Japanese yew (Taxus cuspidata). Tetrahedron, 1996, 52: 12159-12164.
[29] Shigemori H, Sakurai CA, Hosoyama H, et al . Taxezopidines J, K and L, new taxoids from Taxus cuspidata inhibiting Ca2+-induced depolymerization of microtubules. Tetrahedron, 1999, 55: 2553-2558.
[30] Yoshizaki F, Fukuda M, Hisamichi S, et al . Stucture of taxane diterpenoids from the seeds of Japan yew (Taxus cuspidata). Chem Pharm Bull, 1988, 36: 2098-2102.
[31] Nizam UK,Nazneen P. The constituents of genus Taxus. Journal of Scientific & Industrial Research
1987, 46: 512-516.
[32] Beutler JA, Chmurny GM,Look SA, et al. Taxinine M, a new tetracyclic taxane from Taxus brevifolia. J Nat Prod, 1991, 54: 893-897.
[33] Barboni L, Gariboldi P, Torregiani E, et al. Minor taxoids from Taxus wallichiana. J Nat Prod, 1995, 58: 934-939.
[34] Hosoyama H, Inubushi A, Katsui T. Taxuspines U, V and W, new taxanes and related diterpenoids from Japanese yew (Taxus cuspidata). Tetrahedron, 1996, 52: 13145-13150.
[35] Shi QW,Oritani T,Horiguchi T, et al . Four novel taxane diterpenoids from the needles of Japanese yew (Taxus cuspidata).Biosci Biotechnol Biochem, 1999, 63: 924-929.
[36] Wang XX,Shigemori H , Kobayashi J . Taxezopidine A, a novel taxoid from Japanese yew, (Taxus cuspidata). Tetrahedron Lett, 1997, 38: 7587-7588.
[37] Morita Gonda A. Wei LYamamura Y. Takeya Itok Awa. Journal of Natural Products 1997, 46, 583.
[38] Shi QW, Oritani T, Kiyota H, et al. Taxane diterpenoids fromTaxus yunnanensis and Taxus cuspidata. Phytochemistry, 2000, 54: 829-834.
[39] Shi QW, Cao CM, Gu JS , et al. Four new taxane metabolites from the needles of Taxus cuspidata. Nat Prod Res, 2006, 20: 173-179.
[40] Kosugi K, Sakai J, Zhang S, et al. Neutral taxoids from Taxuscuspidata as modulators of multidrug
-resistant tumor cells. Phytochemistry, 2000, 54: 839-845.
[41] Kobayashi J, Hosoyama H, Shigemori H, et al. Taxuspine D, a new taxane diterpene from Taxus cuspidata with potent in hibitory activity against Ca2 + induced depolymerization of microtubule.
Experientia, 1995, 51: 592-595.
[42] Kobayashi J, Ogiwara A, Hosoyama H, et al . Taxuspine A-C, new taxoids from Japan yew Taxus cuspidata inhibiting drug transport activicity of P-glycorprotein in multidrug-resistant cells. Tetrahedron, 1994, 50: 7401-7416.
[43] Wang XX, Shigemori H, Kobayashi J. Taxuspine K, L and M,new taxoids from Japanese yew (Taxus cuspidata). Tetrahedron, 1996, 52: 2337-2342.
[44] Kobayashi J, Hosoyama H, Katsui T, et al. Taxuspine N, O and P new taxoids from Japanese yew Taxus cuspidata. Tetrahedron. 1996, 52: 5391-5396.
[45] Appendino G, Borboni L, Gariboldi P, et al . Revised structure of brevifoliol and some Baccatin Ⅵ derivatives. J Chem Soc, Chem Commun, 1993: 1587-1589.
[46] Barboni L, Gariboldi P, Torregiani EGA, et al. Taxanes from the needles of Taxus wallichiana. Phytochemistry, 1993, 33: 145-150.
[47] Murakami R, Shi QW,Horiguchi , et al. A novel rearranged taxoid from needles of Japanese yew (Taxus cuspidata). BiosciBiotechnol Biochem1999, 63: 1660-1663.
[48] Morita H, Gonda A, Wei L, et al. Taxuspinananes A and B, new taxoids from Taxus cuspidata var nana. J Nat Prod, 1997, 60: 390-392.
[49] Fuji K, Tanaka K, Li B, et al. Novel diterpenoids from Taxus chinensis. J Nat Prod, 1993, 56: 1520-1531.
[50] Rao K V, Johnson J H.occurrence of 2, 6-dimethoxy cinnamaldehyde in Taxus floridana and structural revision of taxiflorine to taxchinin M. Phytochemistry, 1998, 49: 1361-1364.
[51] Morita H, Gonda A, Wei L, et al . Four new taxoids from Taxus cuspidata var. nana. Planta Med, 1998, 64: 183-186.
[52] Barboni L, Gariboldi P, Appendino G, et al. New taxoids fromTaxus baccata. Liebigs Ann, 1995, (3):
345-349.
[53] Shi QW, Oritani T,Hiromasa K, et al . Three new taxoids fromthe leaves of the Japanese yew, Taxus
cuspidata. Nat Prod Lett, 2001, 15: 55-62.
[54] Morita H, Gonda A, Wei L, et al . Taxoids from Taxus cuspidate var. nana. Phytochemistry 1998,
48: 857-862.
[55] Shi QW, Li ZP, Zhao D, et al. New 2 (3 -20) abeotaxanes and 3, 11-cyclotaxane from needles of Taxus cuspidata. BiosciBiotechnol Biochem, 2004, 68: 1584-1587.
[56] Sakai J, Sasaki H, Kosugi K, et al. Two taxoids Taxus cuspidata as modulartors of multidrug resistant tumor cells. Heterocycles, 2001, 54: 999-1009.
[57] Shi QW,Oritani T,Sugiyama T, et al. Three new bicyclic taxane diterpenoids from the needles of Japanese yew ,(Taxus cuspidata). J Asian Nat Prod Res, 1999, 2: 63-70.
[58] Murakami R, Shi QW,Oritani T. A taxoid from the needles of the Japanese yew, Taxus cuspidata. Phytochemistry, 1999, 52: 1577-1580.
[59] Nguyen TD, Banskota AH, Tezuka Y, et al. Diterpenes and sesquiterpenes from the bark of Taxus yunnanensis. Phytochemistry, 2003, 64: 1141-1147.
[60] Ripa PV, Matin EA,Cocciolone SM, et al. Fluctuation of phytoecdysteroids in developing shoots of Tasxus cuspidata. Phytochemistry, 1990, 29 :425-427.
[61] Parmer VS, Jha A ,Bisht KS, et al . Constituents of the yew trees. Phytochemistry 1999, 50: 1267-
1304.
[62] Yoshizaki F, Fufuda M, Hisamichi S. Principal constituents of the seeds of Japan yew (Taxus cuspidata). Shoyakugaku Zashi, 1986, 40: 429.
[63] Erdtman H, Tsuno K. Taxus heartwood constituents. Phytochemistry, 1969, 8: 931-932.
[64] Shi QW taxanes in rooted cutting vs mature Japanese yew Canadian Journal of chemistry 2003. 81(4)
338
[65] 张宏桂，张宏，阎吉昌等。东北红豆杉枝叶化学成分研究 人参研究, 1995, 4.
[66] Kobayashi J, Hosoyama H,Wang XX,et al. Modulation of multi-drug resistance by taxuspine C and other taxoids from Japanese yew. Bioorg Med Chem Lett, 1997, 7: 393-398.
[67] Masahiko Oshige, a, Mika Takenouchi,d Yasutaro Kato,Bioorganic & Medicinal Chemistry 12 (2004) 2597–2601.
[68] 刘海青, 林瑞超, 马双成等。旱生卷柏化学成分的研究 (Ⅰ)。 中草药，2003：34（4）298-299。
[69] 许婧, 李铣, 张鹏等。金刚藤的黄酮类化学成分。沈阳药科大学学报。2004：21（6）424-425。
[70] 穆丽华, 张东明。紫荆化学成分的研究。中国中药杂志2006，3 (21)：1795-1797。
[71] 丁林生，梁侨丽，腾艳芬。枳棋子黄酮类成分研究。药学学报1997，32 (8)：600-602。
[72] 周金云，方起程。天然紫杉烷类二萜类化合物的核磁共振氢谱规律。植物学报1997，39 (5)：467-76。
[73] Yaeko konda .tomomistu Sasaki Absolute structure of 2′ desacetoxyaustrospicatine established by x-ray analysis and circular dichroism spectroscopy . Chem Pharm Bull, 1994: 2621-2623
[74] 霍长虹，李作平，史清文等。天然紫杉烷类二萜类化合物的核磁共振氢谱特征。波谱学杂志 2007，24 (1)：101-118。
[75] 梅文莉, 曾艳波, 丁中涛等。红树植物卤蕨化学成分的分离与鉴定。中国药物化学杂志。2006： 46-48。
[76] 张君增，方起程，梁哓天。中国特有植物白豆杉的化学成分研究。植物学报1996，38 (5)：399-340。
[77] 肖海涛, 李铣。半枝莲的化学成分。沈阳药科大学学报 2006，23 (10)：637-639。
[78] 汤海峰 ，易杨华 ，姚新生等。褐藻果叶马尾藻化学成分的研究《中国海洋药物》2002，6：11-15。
作者点评：
1. 近年来，科学家研究发现一些紫杉烷类化合物虽然其本身没有抗肿瘤活性，但可以作为多药耐药逆转剂改变耐药细胞的耐药性。如紫杉宁及其衍生物，此类化合物具有较高的逆转多药耐药活性，并且具有很低的细胞毒性，这对于将其发展成为新型多药耐药逆转剂是很有利的，因紫杉宁在东北红豆杉枝叶中含量较高，可利用其做为先导化合物进行结构修饰，以便寻找具有较高的逆转多药耐药活性的目标化合物。
2 用开放硅胶柱色谱和凝胶柱色谱及重结晶等技术对东北红豆杉枝叶75%乙醇冷浸提取物进行分离，分离得到17个化合物。根据化合物的理化性质和光谱数据确定了15个化合物的结构，分别为紫杉双黄酮 A (1)、穗花杉双黄酮 (2)、槲皮素 (3)、山柰酚 (4)、紫杉叶素 (5)、二氢山柰酚 (6)、紫杉宁 (7)、紫杉宁A (8)、紫杉宁B (9)、taxacin (10)、坡那甾酮A (11)、胡萝卜苷 (12)、红豆杉苷 (13)、对羟基苯甲醛 (14)、对羟基苯甲酸 (15)。其中化合物紫杉双黄酮 A为一未见文献报道的新化合物，化合物紫杉叶素、二氢山柰酚、对羟基苯甲酸、对羟基苯甲醛为首次从该植物中分离得到，化合物taxacin利用二维核磁共振技术首次对其氢碳数据进行了全归属。
3 传统中医药中以红豆杉属植物东北红豆杉T. cuspidata的枝叶入药，中药处方名紫杉、赤柏松、紫柏松。《中药大辞典》记载：红豆杉中含有的生物碱有利尿、通经、降压、降糖、杀虫、止痒作用，对治疗肾脏病、糖尿病、风湿病和白血病等均有良好疗效。也可从中医药的角度对其药效作用的物质基础进行研究。