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冬凌草和柔茎香茶菜的化学成分及生物活性研究
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摘要
本论文由三章组成。前两章分别详细论述了冬凌草(Isodon rubescens)和柔茎香茶菜(I. flexicaulis)的化学成分及生物活性研究。论文的第三章为2009年1月至2012年10月这段时间内二萜生物碱研究进展的综述。
     综合利用各种分离技术、现代波谱学方法和晶体X-射线衍射等手段,从上述三种植物中共鉴定了125个/次化合物,53个为新化合物。已鉴定的化合物类型涉及单萜、倍半萜、二萜、三萜、生物碱、木脂素、甾醇、黄酮和其它酚性成分。新化合物的类型属于二萜及其含氮衍生物,首次从香茶菜属植物中发现了一种新类型的二萜生物碱——Kaurine,这种生物碱结构上具有对映-贝壳杉烷二萜的骨架,又不同于以往发现的典型二萜生物碱,活性上的表现也不同于对映-贝壳杉烷二萜。此外,从冬凌草中我们还发现了4个少见的11,20-环氧-对映-贝壳杉烷二萜化合物,其中的1个具有细胞毒活性的化合物提示了可能存在不同于α-亚甲基环戊酮的药效团。从柔茎香茶菜中,我们还首次发现了天然存在的13,14-开环-松香烷二萜flexicaulin E。这些具有新颖结构和生物活性的二萜及其类似物的发现,加深了对香茶菜属植物化学成分的认识,同时为进一步合理开发该属植物资源提供了新的化学物质基础。
     第一章冬凌草的化学成分和生物活性研究
     冬凌草(Isodon rubescens)系唇形科香茶菜属草本植物,在河南省一带主要用于治疗各种炎症和消化道癌等疾病,以之为原料的抗癌辅助药物冬凌草片、治疗咽喉肿痛的冬凌草含片等制剂早已上市,其主要活性物质为以冬凌草甲素和乙素为代表的一系列对映-贝壳杉烷二萜。中科院昆明植物研究所孙汉董研究组系统地研究了产自河南、河北以及贵州的多个县市地区的冬凌草,发现其化学成分常随生境的不同而有较大差异,这种差异性甚至超过了同属不同种间的差异。基于化学成分多样性和发现生物活性化合物的目的,我们对产自湖北省建始县的冬凌草地上部分的化学成分进行了详细的研究,从其95%的乙醇提取物的乙酸乙酯萃取物中分离鉴定了98个化合物,其中二萜类91个,包括47个新二萜,涉及14种不同的结构类型,以7,20-环氧-对映-贝壳杉类型二萜为主。
     第二章柔茎香茶菜的化学成分研究
     柔茎香茶菜(Isodon flexicaulis)系唇形科香茶菜属植物,灌木,主产于四川西南部。鉴于前人对该植物的化学成分及生物活性报道较少,且不同产地的香茶菜属植物次生代谢产物也有所不同,同时为了寻找潜在的抗癌、抗炎活性成分,我们对产自四川省甘孜州道孚县的柔茎香茶菜地上部分的次生代谢成分进行了研究,从70%丙酮提取物的乙酸乙酯部分共分离鉴定了27个化合物(其中新的二萜6个),主要涉及结构类型包括迷迭香酸类、二萜和三萜。
     第三章综述:二萜生物碱研究进展
     本章对2009年1月至2012年10月发表的172个二萜生物碱的结构、名称、来源和参考文献进行了综述,其中包括114个狭义的生物碱,涉及C20-、C19-、C18-和二聚体四大类中的14个小类型,还包括了多种骨架的58个非经典的二萜生物碱。我们对这些化合物的结构类型、名称和来源进行了详尽的总结,区分了狭义的和非典型的二萜生物碱。
This dissertation is composed of three chapters. The chemical and bioactive constituents of Isodon rubescens and I. flexicaulis have been phytochemically studied in chapters1and2, respectively. Chapter3is a review about diterpenoid alkaloids (DAs) reported from2009.1to2012.10.
     On the basis of modern chromatography methods, extensive spectroscopic evidences and single-crystal X-ray diffraction analysis,125compounds, including53new ones, were isolated and identified from the above-mentioned three plants. Types of these compounds include monoterpenoids, sesquiterpenoids, diterpenoids, triterpenoids, alkaloids, lignans, steroids, flavanones, and other phenolic compounds, and new compounds all belong to diterpenoids or their nitrogen-containing derivatives. A new type of DAs, named Kaurine, possess ent-kaurane skeleton and was quite different from all typical DAs in their structures. Unexpectedly, noncytotoxicity of the new diterpenoid alkaloids was quite different from their cytotoxic diterpenoid analogues. Two cytotoxic candidates of four rare11,20-epxoy-ent-kauranoids from I. rubescens suggested the presence of a new cytotoxic center besides a-methylenecyclopentanone. Moreover, flexicaulin E from I. flexicaulis was the first example of natural13,14-seco-abietane.
     Chapter1. Chemical and Bioactive Constituents of Isodon rubescens
     Isodon rubescens (Labiatae) is a perennial herb well known as a folk medicine used in the treatment of inflammation and digestive tract cancer in Henan Province. The standard extract of this plant was successfully developed into a drug product for the treatment of sore throats and inflammation in China. Many investigations on this plant showed that ent-kaurane diterpenoids, exemplified by oridonin and ponicidin, were the major bioactive constituents. Professor Han-Dong Sun and his research group from Kunming Institute of Botany, Chinese Academy of Science, investigated on the constituents of I, rubescens growing in many different regions of Henan, Hebei, and Guizhou Provinces. They found that the structural types of the secondary metabolites changed with different ecological environments for this plant so much that the variability of this species in different places was as much as, or even more significant than that of two species of a same genus. In our present study,97compounds were isolated from the leaves and stems of I. rubescens collected from Jianshi County, Hubei Province. Among them,91variable diterpenoids, including47new ones, could be divided to14subtypes.
     1. Kaurine A (1)*
     2. Kaurine B (2)*
     3. Kaurine C (3)*
     4. Jianshirubesin A (4)*
     5. Jianshirubesin B (5)*
     6. Jianshirubesin C (6)*
     7. Epinodosin (7)
     8. Rabdosin A (8)
     9. Enmein (9)
     10. Rabdosichuanin C (10)
     11. Taibaijaponicain A (11)
     12. Maoyecrystal K (12)
     13. Isodocarpin (13)
     14.6β,15α-Dihydroxy-6,7-seco-6,20-epoxy-1a,7-olide-ent-kaur-16-ene (14)
     15. Epinodosinol (15)
     16.6α,15α-Dihydroxy-20-aldehyde-6,7-seco-6,11a-epoxy-ent-kaur-16-en-1a,7-olide (16)
     17. laxiflorin C (17)
     18. laxiflorin D (18)
     19. Laxiflorin E (19)
     20. Jianshirubesin D (20)*
     21. Jianshirubesin E (21)*
     22. Jianshirubesin F (22)*
     23. Rubescensin W (23)
     24. Hubeirubesin A (24)*
     25. Hubeirubesin B (25)*
     26. Jianshirubesin G (26)*
     27. Jianshirubesin H (27)*
     28. Jianshirubesin I (28)*
     29. Ponicidin(29)
     30. Hubeirubesin C (30)*
     31. Hubeirubesin D (31)*
     32. Hubeirubesin E (32)*
     33. Hubeirubesin F (33)*
     34. Hubeirubesin G (34)*
     35. Hubeirubesin H (35)*
     36. Hubeirubesin I (36)*
     37. Hubeirubesin J (37)*
     38. Hubeirubesin K (38)*
     39. Hubeirubesin L (39)*
     40. Hubeirubesin M (40)*
     41. Hubeirubesin N (41)*
     42. Hubeirubesin O (42)*
     43. Hubeirubesin P (43)*
     44. Hubeirubesin Q (44)*
     45. Hubeirubesin R (45)*
     46. Hubeirubesin S (46)*
     47. Hubeirubesin T (47)*
     48. Hubeirubesin U (48)*
     49. Hubeirubesin V (49)*
     50. Hubeirubesin W (50)*
     51. Hubeirubesin X (51)*
     52. Hubeirubesin Y (52)*
     53. Hubeirubesin Z (53)*
     54. Jianshirubesin J (54)*
     55. Jianshirubesin K (55)*
     56. Jianshirubesin L (56)*
     57. Jianshirubesin M (57)*
     58. Oridonin (58)
     59. Rubescensin C (59)
     60. Rubescensin E (60)
     61. Rubescensin G (61)
     62. Rubescensin Q (62)
     63.6β,7β,14β,15β-Tetrahydroxy-7a,20-epoxy-ent-kaur-16-ene (63)
     64. Maoecrystal X (64)
     65. Maoyecrystal F (65)
     66. Acetonide of maoyecrystal F (66)
     67. Hebeirubescensin K (67)
     68. Wikstroemioidin B (68)
     69. Rabdoternin A (69)
     70. Rabdoternin B (70)
     71. Rabdoternin D (71)
     72. Rabdoternin F (72)
     73. Shikokianin (73)
     74. Lasiodin (74)
     75. Lasiokaurinol (75)
     76. Enmenin (76)
     77. Enmenin monoacetate (77)
     78. Rabdolongin A (78)
     79. Parvifoline F (79)
     80. Odonicin (80)
     81. Parvifoline AA (81)
     82. ent-Abierubesin A (82)*
     83. ent-Abierubesin B (83)*
     84. ent-Abierubesin C (84)*
     85. ent-Abierubesin D (85)*
     86. ent-Abierubesin E (86)*
     87. Hebeiabinin B (87)
     88. ent-Abienervonin C (88)
     89. Rabdoepigibberellolide (89)
     90. Lushanrubescensin J (90)
     91. Phytol(91)
     92. Tormentic acid (92)
     93. Heterobetulinic acid (93)
     94. Pinoresinol (94)
     95. Acetoxypinoresinol (95)
     96.5,6-Dihydroxy-7,3',4'-trimethoxyflavone (96)
     97. β-Sitosterol (97)
     98. Blumenol(98) NOTE:*new compound
     Chapter2. Chemical Constituents of Isodon flexicaulis
     Isodon flexicaulis belongs to the genus Isodon of the family Labiatae. It is a shrub mainly distributed in the southwest of Sichuan Province and has not been well-studied in fields of phytochemistry and pharmacology. In order to search new bioactive constituents, the arieal parts of I. flexicaulis collected in Daofu County, Sichuan Province, were systematically investigated, and27compounds, including six new diterpenoids, were isolated and identified. The types of these compounds are mainly the rosmaric acid, diterpenoid, and triterpenoid.
     1. Flexicaulin B (1)*
     2. Flexicaulin C (2)*
     3. Flexicaulin D (3)*
     4. Forrestin B (4)
     5. Tenuifolin F4(5)
     6. Inflexarabdonin D (6)
     7. Flexicaulin E (7)*
     8. Flexicaulin F (8)*
     9. Flexicaulin G (9)*
     10.7α-Hydroxydehydroabietic acid (10)
     11.15-Hydroxydehydroabietic acid (11)
     12. Pomiferin F(12)
     13. Rotundic acid (13)
     14. Tormentic acid (14)
     15. Corsolic acid (15)
     16.23-Hydroxyursolic acid (16)
     17. Maslic acid (17)
     18. Teuclatriol (18)
     19. menth-l-ene-3,6-diol (19)
     20.1-Hydroxypinoresinol (20)
     21. Epipinoresinol (21)
     22. Lariciresinol (22)
     23. Caffeic acid (23)
     24. Rosmarinic acid (24)
     25.3'-O-Methyl-rosmarinic acid (25)
     26. Methyl isoferuloyl-7-(3,4-dihydroxyphenyl)-lactate (26)
     27. Methyl-6-dehydroxyl-rosmarinate (27)
     NOTE:*new compound
     Chapter3. Diterpene Alkaloids (2009.01-2012.10)
     172diterpene alkaloids (DAs) reported from2009.01to2012.10were reviewed. These compounds involved114typical DAs belonging to C20-, C19-, C18-, and dimeric four types and more than10subtypes, as well as another58untypical DAs. In this review, structural types, compound names, and plant sources are summarized in detail so as to distinguish typical and untypical DAs.
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