臺灣博碩士論文加值系統

本研究乃是將豆科植物，野百合屬太陽麻 (Crotalaria juncea L.)進行成分分離純化並進行生物活性探討。台灣產太陽麻2004年6月採於屏東縣，取莖部350 g，室溫下以甲醇冷浸約一星期後，將其抽出液經減壓濃縮後之抽出物50 g。以矽膠柱層層析法分離，共分離得到十四個化合物，其中包含十二個已知化合物分別為β-Sitosterol (1)， β-Sitosterol-3-O-β-D-glucoside (2)，Betulinic acid (3)，β-Amyrin (4)， 22-Episoyasapogenol B (5)，Genistein (6)，Daidzein (7)，7,4?S-Dihydroxyflavone (8)，3?S,4?S-Dihydroxy-7-methoxyflavone (9)，3?S,4?S-Dihydroxy-5,7-methoxyflavone (10)，Crotafuran A (13)及 Crotafuran B (14)。另有二個新化合物為 Crotafuran F (11) 及Crotafuran G (12)。
將化合物4進行抗發炎 (anti-inflammatory effect) 生物活性之篩選，結果顯示以PMA (3 nM)所誘發老鼠嗜中性白血球形成超氧陰離子 (superoxide anion generation)的作用中，呈現抑制的活性且抑制能力與濃度相關，其IC50值為15.9 ?b 2.3 µM。

With the scope of our search for biologically active compounds from plants, twelve known compounds, β-Sitosterol (1), β-Sitosterol-3-O-β-D-
glucoside (2), Betulinic acid (3), β-Amyrin (4), 22-Episoyasapogenol B (5), Genistein (6), Daidzein (7), 7,4?S-Dihydroxyflavone (8), 3?S,4?S- Dihydroxy-7-methoxyflavone (9), 3?S,4?S-Dihydroxy-5,7-methoxyflavone (10), Crotafuran A (13), and Crotafuran B (14), and two new compounds, Crotafuran F (11) and Crotafuran G (12), were isolated from the stem of Crotalaria juncea L.. Their structures were determined by spectroscopic methods and comparison with those data reported in literatures.
Compound 4 showed significant inhibitory effects on the superoxide anion generation in rat neutrophils stimulated with PMA (3 nM) in a concentration-dependent manner with an IC50 value of 15.9 ?b 2.3 µM.

目錄
中文摘要……………………………………………………………………………...1
英文摘要…………………………………………………………………………….2
第一章 研究背景與目的……………………………………………………………3
第二章 臺產野百合屬植物—太陽麻之化學成份研究…………………………..7
第一節 植物型態……………………………………………………………..11
第二節 抽取與分離…………………………………………………………….12
第三節 化合物結構之鑑定
第一項 β-Sitosterol (1) 的結構式證明………………………………………….16
第二項 β-Sitosterol-3-O-β-D-glucoside (2)的結構式證明…………………..19
第三項 Betulinic acid (3) 的結構式證明…………………………………….23
第四項β-Amyrin (4) 的結構式證明………………………………………26
第五項 22-Episoyasapogenol B (5) 的結構式證明…………………………29
第六項 Genistein (6) 的結構式證明………………………………………33
第七項 Daidzein (7) 的結構式證明…………………………………………….36
第八項 7,4?S-Dihydroxyflavone (8) 的結構式證明…………………………39
第九項 3?S,4?S-Dihydroxy-7-methoxyflavone (9) 的結構式證明…………….42
第十項 3?S,4?S-Dihydroxy-5,7-methoxyflavone(10)的結構式證明…………..46
第十一項 Crotafuran F (11) 的結構式證明…………………………………50
第十二項 Crotafuran G (12) 的結構式證明…………………………………57
第十三項Crotafuran A (13) 的結構式證明………………………………63
第十四項 Crotafuran B (14)的結構式證明………………………………….65
第三章 抗發炎活性試驗結果……………………………………………………..67
第四章 結果與討論……………………………………………………………….70
第五章 實驗部份
第一節 儀器與材料……………………………………………………………73
第二節 實驗方法……………………………………………………………….76
第三節 生物活性試驗篩選方法………………………………………………..77
第四節 化合物之物理化學性質………………………………………………..81
參考文獻……………………………………………………………………………95
圖表目錄
圖1-1 β-Sitosterol (1) 紅外線光譜………………………………………………..17
圖1-2 β-Sitosterol (1) 氫光譜………………………………………………………17
圖1-3 β-Sitosterol (1) 碳光譜……………………………………………………….????
圖1-4 β-Sitosterol (1) 質譜………………………………………………………….18
圖 2-1 β-Sitosterol-3-O-β-D-glucoside (2) 紅外線光譜………………………….20
圖 2-2 β-Sitosterol-3-O-β-D-glucoside (2) 氫光譜……………………………….21
圖 2-3 β-Sitosterol-3-O-β-D-glucoside (2) 碳光譜...................................................21
圖 2-4 β-Sitosterol-3-O-β-D-glucoside (2) 質譜…………………………………..22
圖3-1 Betulinic acid (3) 紅外線光譜………………………………………………24
圖3-2 Betulinic acid (3) 氫光譜…………………………………………………..24
圖 3-3 Betulinic acid (3) 碳光譜……………………………………………………25
圖 3-4 Betulinic acid (3) 質譜………………………………………………………25
圖4-1 β-Amyrin (4) 紅外線光譜……………………………………………………27
圖4-2 β-Amyrin (4) 氫光譜…………………………………………………………27
圖4-3 β-Amyrin (4) 質譜……………………………………………………………28
圖5-1 22-Episoyasapogenol B (5) 紅外線光譜…………………………………….30
圖5-2 22-Episoyasapogenol B (5) 氫光譜…………………………………………31
圖5-3 22-Episoyasapogenol B (5) 碳光譜…………………………………………31
圖5-4 22-Episoyasapogenol B (5) 質譜…………………………………………..32
圖6-1 Genistein (6) 紅外線光譜…………………………………………………..34
圖6-2 Genistein (6) 氫光譜………………………………………………………..34
圖6-3 Genistein (6) 質譜…………………………………………………………35
圖7-1 Daidzein (7) 紅外線光譜……………………………………………………….37
圖7-2 Daidzein (7) 氫光譜……………………………………………………………37
圖7-3 Daidzein (7) 質譜………………………………………………………………..38
圖8-1 7,4?S-Dihydroxyflavone (8) 紅外線光譜…………………………………….40
圖8-2 7,4?S-Dihydroxyflavone (8) 氫光譜…………………………………………..41
圖8-3 7,4?S-Dihydroxyflavone (8) 質譜……………………………………………..41
圖9-1 3?S,4?S-Dihydroxy-7-methoxyflavone (9) 紅外線光譜………………………..43
圖9-2 3?S,4?S-Dihydroxy-7-methoxyflavone (9) 氫光譜……………………………..44
圖9-3 3?S,4?S-Dihydroxy-7-methoxyflavone (9) DEPT光譜…………………………44
圖9-4 3?S,4?S-Dihydroxy-7-methoxyflavone (9) 質譜………………………………45
圖10-1 3?S,4?S-Dihydroxy-5,7-methoxyflavone (10) 紅外線光譜………………….47
圖10-1 3?S,4?S-Dihydroxy-5,7-methoxyflavone (10) 氫光譜…………………………48
圖10-1 3?S,4?S-Dihydroxy-5,7-methoxyflavone (10) 碳光譜…………………………48
圖10-1 3?S,4?S-Dihydroxy-5,7-methoxyflavone (10) 質譜……………………..……..49
圖11-1 Crotafuran F (11) 紅外線光譜……………………………………………..53
圖11-2 Crotafuran F (11) 氫光譜…………………………………………………..????
圖11-3 Crotafuran F (11) 碳及DEPT光譜…………………………………………54
圖11-4 Crotafuran F (11) HMQC光譜…………………………………………..54
圖11-5 Crotafuran F (11) HMBC光譜………….……………………………….55
圖11-6 Crotafuran F (11) COSY光譜……………………………………………….55
圖11-7 Crotafuran F (11) NOESY光譜……………………………………………56
圖11-8 Crotafuran F (11) 質譜…………………………………………………….56
圖12-1 Crotafuran G (12) 紅外線光譜……………………………………………59
圖12-2 Crotafuran G (12) 氫光譜…………………………………………………60
圖12-3 Crotafuran G (12) DEPT光譜………………………………………………60
圖12-4 Crotafuran G (12) HMQC光譜…………………………………………….61
圖12-5 Crotafuran G (12) HMBC光譜.......................................................................61
圖12-6 Crotafuran G (12) COSY光譜…………………………………………….62
圖12-7 Crotafuran G (12) 質譜…………………………………………………….62
圖13-1 Crotafuran A (13)氫光譜……………………………………………………64
圖14-1 Crotafuran B (14)氫光譜………………………………………………….66
Table 1. Constituents Isolated from Crotalaria………………………………………..7
Table 2. The inhibitory effects of 4 on the release of β-glucuronidase and histamine from rat peritoneal mast cells stimulated with compound 48/80…………………..68
Table 3. The inhibitory effects of 4 on the release of β-glucuronidase and lysozyme from rat neutrophils stimulated with fMLP/CB……………………………………68
Table 4. The inhibitory effects of 4 on superoxide anion generation in rat neutrophils stimulated with fMLP (0.3 µM)/CB (5 µg mL ) or PMA (3 nM)…………………69
Table 5. The inhibitory effects of 4 on the NO production in RAW 264.7 Cells stimulated with LPS (1 µg/ml)……………………………………………………..69

1. O. Hiroyoshi. Leguminosae in Flora of Taiwan. 1. ed. Editorial Committee of Flora of Taiwan: Taipei, 1993; Vol III: 217.
2. O. Hiroyoshi. Leguminosae in Flora of Taiwan. 1. ed. Editorial Committee of Flora of Taiwan: Taipei, 1993; Vol III: 228.
3. B. Abegaz, G. Atnafu, H. Duddeck, and G. Snatzke, Tetrahedron, 1987, 43, 3263.
4. D. L.Cheng, S. B. Tu, A. A. Enti, and E. Roeder, Sci. Pharm., 1986, 54, 351. [Chem. Abst., 1987, 106, 17293c].
5. Group Crotalaria Plants Res., Hupeh Inst. Of Botany, Chih Wu Hsueh Pao, 1974, 16, 380. [Chem. Abst., 1975, 83, 25052v].
6. A. R. Mattocks, and N. Nwude, Phytochemistry, 1988, 27, 3289.
7. J. A. Edgar, H. J. Lin, C. R. Kumana, and M. M. T. Ng, Am. J. Chin. Med., 1992, 100, 281.
8. R. Adams, and B. L. Van Duuren, J. Am. Chem. Soc., 1953, 75, 4631.
9. C. C. J. Culvenor, and L. W. Smith, Aust. J. Chem., 1959, 12, 255.
10. C. C. J. Culvenor, and L. W. Smith, Aust. J. Chem., 1962, 15, 328.
11. M. L. Sethi, and C. K. Atal, Planta Med., 1964, 12, 173.
12. C. K. Atal, K. K. Kapur, C. C. J. Culvenor, and L. W. Smith, Tetrahedron Lett., 1966, 537.
13. D. S. Bhakuni, R. Chaturvedi, J. Nat. Prod., 1984, 47, 585.
14. C. C. W. Wanjala, and R. R. T. Majinda, Phytochemistry, 1999, 51, 705.
15. J. R.Weng, M. H. Yen, and C. N. Lin, Helv. Chim. Acta, 2002, 85, 847.
16. L. W. Smith, and C. C. J. Culvenor, J. Nat. Prod., 1981, 44, 129.
17. R. N. Yadava, and Abha Singh, Fitoterapia, 1993, 276.
18. K. Brown, J. A. Devlin, and D. J. Bobins, Phytochemistry, 1984, 23, 457.
19. H. Wiedenfeld, E. Roeder, and E. Anders, Phytochemistry, 1985, 24, 376.
20. G. Toppel, L. Witte, and T. Hartmann, Phytochemistry, 1988, 27, 3757.
21. A. K. Gupta, J. N. Bemiller, Phytochemistry, 1990, 29, 853.
22. R. N. Gandhi, T. R. Rajagopalan, and T. R. Seshardi, Curr. Sci. 1966, 35, 514. [Chem. Abst., 1967, 66, 10080].
23. M. A. Ali, and Z. Kapadia, Pakistan J. Sci. Ind. Res., 1967, 10, 80. [Chem. Abst., 1968, 68, 57385].
24. S. S. Subramanian, and S. Nagarajan, Planta Med., 1968, 16, 432.
25. R. S. Swahney, and C. K. Atal, J. Indian Chem. Soc., 1968, 45, 1052.
26. K. Suri, O. P. Suri, K. L. Dhar, and C. K. Atal, J. Indian Chem., Sect. B, 1978, 16 (B), 78.
27. P. Hoet, M. Astigueta, and A. M. Frisque, Rev. Latinoam. Quim., 1981, 12, 34. [Chem. Abst., 1981, 94, 188670].
28. R. Manavalan, and B. M. Mithal, Indian Drugs, 1982, 19, 385.
29. D. Prakash, S. P. Wahi, and S. Ghosal, Indian Drugs, 1985, 22, 495.
30. M. Manzoori-i-Khuda, A. K. Chowdhury, T. Reza, S. A. Chowdhury, Bangladesh J. Sci. Ind. Res., 1986, 21, 40. [Chem. Abst., 1989, 111, 4206].
31. Z. Z. Ibraheim, Bull. Fac. Sci., Assiut Univ., B, 1994, 23, 49. [Chem. Abst., 1996, 124, 50617].
32. S. W. Yang, G. A. Cordell, H. Lotter, H. Wagner, B. C. Mouly, A. V. N. A. Rao, and P. S. Rao, J. Nat. Prod., 1998, 61, 1274.
33. K. C. Naidu, V. Aruna, and T. Satyanarayana, Indian J. Nat. Prod., 1998, 14, 17.
34. M. A. Ramadan, Z. Z. Ibraheim, A. M. Abdel-Baky, D. W. Bishay, and H. Itokawa, Bull. Pharm. Sci. Assiut Univ., 1999, 22, 131.
35. Y. Ding, J. kinjo, C. Yang, and T. Nohara, Chem. Pharm. Bull., 1991, 39, 496.
36. D. J. Pilbeam, A. J. Lyon-Joyce, and E. A. Bell, J. Nat. Prod., 1983, 46, 601.
37. M. L. Sharma, G. B. Singh, and B. J. Ray Ghatak, Indian J. Exp. Biol., 1967, 5, 149.
38. G. B. Singh, M. L. Sharma, and B. J. Ray Ghatak, Indian J. Exp. Biol., 1969, 7, 144.
39. S. Snehalata, and M. N. Ghosh, Indian J. Med. Res., 1968, 59, 1386.
40. M. L. Sharma, and B. J. Ray Ghatak, Indian J. Exp. Biol., 1971, 9, 177.
41. J. R.Weng, L. T. Tsao, M. H. Yen, J. P. Wang, and C. N. Lin, J. Nat.Prod., 2003, 66, 404.
42. H. H. Ko, J. R. Weng, L.T. Tsao, M. H. Yen, J. P. Wang, and C. N. Lin, Bioorg. Med. Chem. Lett., 2004, 14, 1011.
43. M. D. Khalilulah, V. M. Sharma, P. S. Rao, and K. Raghavaraju, J.Nat.Prod.,1992, 55, 229.
44. M. Abdul, I. Hiroko, S. Mariko, N. Osamu, O. Tetsuo, Cytotechnology, 2003, 43, 127.
45. S. Moncada, R. M. Palmer, and E. A. Higgs, Pharmacol. Rev., 1991, 43, 109.
46. R. Adams and M. Gianturco, J. Am. Chem. Soc., 1956, 78, 1919.
47. E. H. M. Mendonça, P. Mazzafera, and M. A. Schiavinato, Phytochemistry, 1999, 50, 313.
48. D. Nobre, M. L. Dagli, and M. Haraguchi, Vet. Hum. Toxicol., 1994, 36, 445.
49. A. O. Prakash, S. Dehadrai, and S. Jonathan, J. Ethnopharmacol., 1995, 45, 167.
50. Y. Fukuyama, A. Komiyama, Y. Mima, and M. Kodama, Phytochemistry, 1991, 30, 3433.
51. H. Minami, E. Takahashi, H. Fukuyama, M. Kodama, T. Yoshizawa, and K. Nakagawa, Chem. Pharm. Bull., 1995, 43, 347.
52. M. M. Gutpa, and Y. N. Shukda, Phytochemistry, 1986, 25, 1423.
53. A. M. Iribarren, and A. B. Pomilio, Phytochemistry, 1982, 21, 2107.
54. H. J. Reich, M. Jautelat, and M. T. Messe, J. Am. Chem. Soc., 1990, 91, 7445.
55. H. Budzikiewicz, J. M. Wilson, and C. Djerassi, J. Am. Chem. Soc., 1963, 85, 3688.
56. S. Seo, and Y. Tomita, Tetrahedron Lett., 1975, 1, 7.
57. J. B. Harborne, “The Flavonoids” chap. 2 eds. By J. B. Harborne, T. J. Mabry, and H. Mabry, Champman & Hall, London, 1975, 47.
58. J. B. Harborne, “The Flavonoids” chap. 3 eds. By J. B. Harborne, T. J. Mabry, and H. Mabry, Champman & Hall, London, 1975. 97.
59. A. Pelter, R. S. Ward, and R. J. Bass, J. Chem. Soc. Perkin Trans 1, 1978, 666.
60. P. K. Agrawal, “ Carbon-13 NMR of Flavonoids” chap. 4 eds. By P. K. Agrawal, New York, USA, 1989. 29.
61. H. L. Jha, F. Zilliken, and E. Breitmaier, Can. J. Chem. 1980, 58, 1211.
62. L. A. Mitscher, S. R. Gollapudi, D. C. Gerlach, S. D. Drake, E. A. Véliz, and J. A. Ward, Phytochemistry, 1988, 27, 381.
63. K. Biemann, “Spectral Data for Structure Determination of Organic Compounds”, Springer-Verlag, Cambridge, MA 02139/USA, 1989, H265.
64. 翁靖如，私立高雄醫學大學藥學研究所，博士論文，2002。
65. F. A. A. V. Acker, J. A. Hageman, G. R. M. M. Haenen, W. J. F. Vijgh, A. Bast and W. M. P. B. Menge, J. Med. Chem, 2000, 43, 3752.
66. J. Wen, H. M. Shi and P. F. Tu, Biochem. syst. ecol. , 2006, 34, 177.
67. T. HIJWEGEN, Phytochcmistry, 1973, 12, 375.