本論文以人類嗜中性白血球細胞(human neutrophil)進行超氧陰離子生成(superoxide anion generation)及彈性蛋白酶釋放(elastase release)的抗發炎活性測試去解釋印尼血竭(Daemonorops draco)的傳統藥效，並以其分離之劃分層的活性測試結果做為分離指標 。印尼血竭分離得到總計59個化合物，由各種光譜鑑定出5種不同類型之新骨架化合物[dragonbloodin系列共16個(dragonbloodin A1, B1, C1R, C1S, D1R, E1, F1, A2, B2, C2R, C2S, D2R, E2, F2, G2; dragonin A)、2個flavan-chalcone typeⅠ(dragonin B, C)、2個flavan-chalcone typeⅡ(dragonin D, E)、3個flavan-quinone (dragonin F1, F2, G)、1個flavan-lignol (dragonlignin B)]、16個新化合物[nordracorhodinol; 2,4,6-trihydroxy-3-methyldihydrochalcone; 4,4′-dihydroxy-2,6-methoxy-3-methyldihydrochalcone; draconisin (A, B, D, E, F); sanguisin (A, B); dracoflavan (E1, E2, F1, F2, G); dragonlignin A]和3個天然界首次分離得到之化合物[(2S)-7-methoxyflavan-5-ol; draconisin C; (−)-draconol]。另外，16個已知化合物經由文獻比對鑑定構造，有6個為首次從黃藤屬植物分離得到之[(−)-alpinetin; (+)-pinoresinol; (+)-syringaresinol; 3′,4′,5-trimethyl-(+)-catechin; 4,6-dihydroxy-2-methoxydihydrochalcone; 1-(2-phenylcarbonyloxyacetyl) benzene]。在活性測試的40個化合物中，兩個flavan-chalcone typeⅠ的新骨架化合物、一個新二聚物以及一對含量較多的立體異構物－dracoflavan B1與B2，對超氧陰離子生成與彈性蛋白酶釋放均有不錯的活性(IC50 = 1.2-3.1 M and 1.0-5.1 M)，另外還有10個化合物也呈現出抗發炎活性(IC50 = 4.0-7.4 M and 3.8-6.5 M)。本研究找出幾個具備抗發炎活性的代表性化合物，期望能開發成抗發炎等新藥。

關鍵字：
印尼血竭；抗發炎活性；超氧陰離子；彈性蛋白酶；新骨架化合物

This study focused on the anti-inflammatory activities of Daemonorops draco by evaluating superoxide anion generation and elastase release of human neutrophil. By the guide of anti-inflammatory activity, fifty-nine compounds were isolated from Daemonorops draco, including five different types of compounds with unprecedented skeletons [sixteen dragonbloodin series (dragonbloodin A1, B1, C1R, C1S, D1R, E1, F1, A2, B2, C2R, C2S, D2R, E2, F2, G2; dragonin A), two flavan-chalcone typeⅠ(dragonin B, C), two flavan-chalcone typeⅡ(dragonin D, E), three flavan-quinone (dragonin F1, F2, G), and one flavan-lignol (dragonlignin B)], sixteen new compounds [nordracorhodinol; 2,4,6-trihydroxy-3-methyldihydrochalcone; 4,4′-dihydroxy-2,6-methoxy-3-methyldihydrochalcone; draconisin (A, B, D, E, F); sanguisin (A, B); dracoflavan (E1, E2, F1, F2, G); dragonlignin A], and three compounds separated from natural sources first time [(2S)-7-methoxyflavan-5-ol; draconisin C; (−)-draconol]. The chemical structure of these compounds were confirmed via 1D-NMR, 2D-NMR, X-ray/SCD, IR, UV, Polarization, as well as ECD spectrum. In addition, sixteen known compounds were identified by literature comparison, and six of them were separated from Daemonorops species first time [(−)-alpinetin; (+)-pinoresinol; (+)-syringaresinol; 3′,4′,5-trimethyl-(+)-catechin; 1-(2-phenylcarbonyloxyacetyl) benzene; 4,6-dihydroxy-2-methoxydihydrochalcone]. Among all forty tested compounds, two flavan-chalcone typeⅠcompounds, one new dimer and one pair of abundant stereoisomers, dracoflavan B1 and B2, have revealed the good anti-inflammatory activity about superoxide anion generation and elastase release (IC50 = 1.2-3.1 M and 1.0-5.1 M). Moreover, the other ten compounds have shown anti-inflammatory activity (IC50 = 4.0-7.4 M and 3.8-6.5 M). Taken together, we report some active components from Daemonorops draco for the development of new anti-inflammatory drugs.

Key words:
Daemonorops draco, anti-inflammatory activity, superoxide anion, elastase, unprecedented skeleton compounds

1. 李時珍, 本草綱目 木部 第三十四卷.
2. 陈可冀; 张之南; 梁子钧, 血瘀证与活血化瘀研究. 上海科学技术出版社: 上海, 1990; p 650.
3. Gupta, D.; Bleakley, B.; Gupta, R. K., Dragon's blood: Botany, chemistry and therapeutic uses. J. Ethnopharmacol. 2008, 115 (3), 361-380.
4. 黃智生, 發炎反應－常見文明病的始作俑者. 科學發展 2008, 422.
5. 國家中醫藥管理局中華本草編委會, 中華本草 第八冊 第二十三卷. 上海科学技术出版社: 上海, 1999; p 8‧455.
6. Gera-Utermhaus; Köhler, F. E., Köhler's Medizinal-Pflanzen in naturgetreuen Abbildungen mit kurz erläuterndem Texte : Atlas zur Pharmacopoea germanica. 1 ed.; 1887; Vol. 2, p plate 167.
7. Gibbs, A.; Green, C.; Doctor, V., Isolation and anticoagulant properties of polysaccharides of Typha angustata and Daemonorops species. Thrombosis research 1983, 32 (2), 97-108.
8. Tsai, W. J.; Hsieh, H. T.; Chen, C. C.; Chen, C. F., Studies on the vasoactive-antithrombotic effect of Draconis Resina and its components. J. Chin. Med. 1995, 6 (1), 59–73.
9. Tsai, W. J.; Hsieh, H. T.; Chen, C. C.; Kuo, Y. C.; Chen, C. F., Characterization of the antiplatelet effects of (2S)-5-methoxy-6-methylflavan-7-ol from Draconis Resina. Eur. J. Pharmacol. 1998, 346 (1), 103-110.
10. 李文靜; 杨光, 血竭在眼科的臨床應用. 遼寧中醫藥大學學報 2007, 9 (5).
11. Yi, T.; Chen, H. B.; Zhao, Z. Z.; Yu, Z. L.; Jiang, Z. H., Comparison of the chemical profiles and anti-platelet aggregation effects of two "Dragon's Blood" drugs used in traditional Chinese medicine. J. Ethnopharmacol. 2011, 133 (2), 796-802.
12. Arnone, A.; Nasini, G.; Vajna de Pava, O.; Merlini, L., Constituents of Dragon's Blood. 5. Dracoflavans B1, B2, C1, C2, D1, and D2, New A-Type Deoxyproanthocyanidins. J. Nat. Prod. 1997, 60 (10), 971-975.
13. Choy, C. S.; Hu, C. M.; Chiu, W. T.; Lam, C. S. K.; Ting, Y.; Tsai, S. H.; Wang, T. C., Suppression of lipopolysaccharide-induced of inducible nitric oxide synthase and cyclooxygenase-2 by Sanguis Draconis, a dragon's blood resin, in RAW 264.7 cells. J. Ethnopharmacol. 2008, 115 (3), 455-462.
14. Chang, Y.; Chang, T. C.; Lee, J. J.; Chang, N. C.; Huang, Y. K.; Choy, C. S.; Jayakumar, T., Sanguis draconis, a dragon's blood resin, attenuates high glucose-induced oxidative stress and endothelial dysfunction in human umbilical vein endothelial cells. ScientificWorldJournal 2014, 2014, 423259 (10 pages).
15. Rao, G. S. R.; Gerhart, M. A.; Lee, R. T., III; Mitscher, L. A.; Drake, S., Antimicrobial agents from higher plants. Dragon's blood resin. J. Nat. Prod. 1982, 45 (5), 646-648.
16. 张培华; 羅少军; 汤少明; 肖佐环; 梁杰; 左強; 李瑾, 血竭素對瘢痕成纖維細胞增殖的影響. 廣東醫學院學報 2005, 23 (4), 361-363.
17. Ji, S.; Zhang, G.; Hua, Y.; Jin, X., Sanguis draconis (Daemonorops draco): a case report of treating a chronic pressure ulcer with tunneling. Holist Nurs Pract 2015, 29 (1), 48-52.
18. H., B.; H., J., Ber. Dtsch Chem. Ges. 1943, 76, 751.
19. Xia, M.; Wang, D.; Wang, M.; Tashiro, S.-i.; Onodera, S.; Minami, M.; Ikejima, T., Dracorhodin perchlorate induces apoptosis via activation of caspases and generation of reactive oxygen species. J Pharmacol Sci 2004, 95, 273-283.
20. 夏明鈺; 王敏偉; 王浩然; 田代真一; 池島喬, 血竭素高氯酸盐诱导人宫颈癌HeLa细胞凋亡的机制. 藥學學報 2004, 39 (12), 966-970.
21. Xia, M.; Wang, M.; Tashiro, S. i.; Onodera, S.; Minami, M.; Ikejima, T., Dracorhodin Perchlorate Induces A375-S2 Cell Apoptosis via Accumulation of p53 and Activation of Caspases. Biol. Pharm. Bull. 2005, 28 (2), 226-232.
22. Xia, M. Y.; Wang, M. W.; Cui, Z.; Tashiro, S. I.; Onodera, S.; Minami, M.; Ikejima, T., Dracorhodin perchlorate induces apoptosis in HL-60 cells. J Asian Nat Prod Res. 2006, 8 (4), 335-343.
23. Shen, C. C.; Tsai, S. Y.; Wei, S. L.; Wang, S. T.; Shieh, B. J.; Chen, C. C., Flavonoids isolated from Draconis Resina. Nat. Prod. Res. 2007, 21 (4), 377-380.
24. 王全胜; 张丽晓; 刘建囯; 汪永辉, 血竭素高氯酸鹽對早期糖尿病腎病腎臟損傷防治作用的研究. 中國藥理學通報 2008, 24 (10), 1331-1334.
25. 汪永辉; 王全胜; 刘建囯; 张丽晓; 张彬, 血竭素高氯酸盐抑制高糖诱导的人肾小球系膜细胞结缔组织生长因子的表达. 中國中藥雜誌 2009, 34 (7), 896-899.
26. 韩晓丽; 朱梅紅; 胡建章; 徐囯兴, 血竭素高氯酸鹽促進離體兔角膜基質細胞凋亡的作用. 眼科研究 2009, 27 (9), 755-759.
27. He, Y.; Ju, W.; Hao, H.; Liu, Q.; Lv, L.; Zeng, F., Dracorhodin perchlorate suppresses proliferation and induces apoptosis in human prostate cancer cell line PC-3. J Huazhong Univ Sci Technolog Med Sci. 2011, 31 (2), 215-219.
28. Wang, W.; Olson, D.; Cheng, B.; Guo, X.; Wang, K., Sanguis Draconis resin stimulates osteoblast alkaline phosphatase activity and mineralization in MC3T3-E1 cells. J Ethnopharmacol. 2012, 142 (1), 168-174.
29. Rasul, A.; Ding, C.; Li, X.; Khan, M.; Yi, F.; Ali, M.; Ma, T., Dracorhodin perchlorate inhibits PI3K/Akt and NF-kappaB activation, up-regulates the expression of p53, and enhances apoptosis. Apoptosis. 2012, 17 (10), 1104-1119.
30. Yu, J. H.; Zheng, G. B.; Liu, C. Y.; Zhang, L. Y.; Gao, H. M.; Zhang, Y. H.; Dai, C. Y.; Huang, L.; Meng, X. Y.; Zhang, W. Y.; Yu, X. F., Dracorhodin perchlorate induced human breast cancer MCF-7 apoptosis through mitochondrial pathways. Int. J. Med. Sci. 2013, 10 (9), 1149-1156.
31. Slade, D.; Ferreira, D.; Marais, J. P., Circular dichroism, a powerful tool for the assessment of absolute configuration of flavonoids. Phytochemistry 2005, 66 (18), 2177-2215.
32. Antus, S.; Kurtan, T.; Juhasz, L.; Kiss, L.; Si, M. H.; Majer, Z., Chiroptical Properties of 2,3-Dihydrobenzo[b]furan and Chromane Chromophores in Naturally Occurring O-Heterocycles. CHIRALITY 2001, 13, 493-506.
33. Robertson, A.; Whalley, W. B., The pigments of “dragon's blood” Resin. Part II. A synthesis of dracorhodin. J. Chem. Soc. 1950, 1882-1884.
34. Robertson, A.; Whalley, W. B.; Yates, J., The Pigments of “Dragon’s Blood ” Resins. Part III. The Constitution of Dracorubin. J. Chem. Soc. 1950, 3117-3123.
35. Olaniyi, A. A.; Powell, J. W.; Whalley, W. B., Pigments of dragon's blood resin. VII. Synthesis of (±)-draconol, (±)-O-methyldraconal, (±)-O-methylisodraconol, and derivatives. Structure of dracorubin. J.C.S. Perkin I 1973, 179-184.
36. Hao, Q.; Saito, Y.; Matsuo, Y.; Li, H. Z.; Takashi, T., Three new flavans in dragon's blood from Daemonorops draco. Nat. Prod. Res. 2015, 1-7.
37. Liu, J.; Dai, H. F.; Wu, J.; Zeng, Y. B.; Mei, W. L., Flavanes from Dracaena cambodiana. Z. Naturforsch 2008, 63b, 1407-1410.
38. Liu, J.; Mei, W. L.; Wu, J.; Zhao, Y. X.; Peng, M.; Dai, H. F., A new cytotoxic homoisoflavonoid from Dracaena cambodiana. J Asian Nat Prod Res. 2009, 11 (2), 192-195.
39. Luo, Y.; Wang, H.; Xu, X.; Mei, W.; Dai, H., Antioxidant phenolic compounds of Dracaena cambodiana. Molecules 2010, 15 (12), 8904-8914.
40. Luo Y.; Dai H. F.; Wang H; Mei W. L., Chemical Constituents from Dragonʹs Blood of Dracaena cambodiana. Chin J Nat Med 2011, 9 (2), 112-114.
41. Collins, D. A.; Haworth, F.; Isarasena, K.; Robertson, A., The Pigments of “ Dragon’s Blood ” Resin. Part I. Dracorubin. J. Chem. Soc. 1950, 1876-1881.
42. Cardillo, G.; Merlini, L.; Nasini, G.; Salvadori, P., Constituents of Dragon's blood. Part I. Structure and absolute configuration of new optically active flavans. J. Chem. Soc. C: Organic 1971, 3967-3970.
43. LucioMerlini; Nasini, G., Constituents of Dragon's Blood. Part II. Structure and Oxidative Conversion of a Novel Secobiflavonoid. J.C.S. Perkin I 1976, 1570-1576.
44. Arnone, A.; Nasini, G.; Merlini, L., Constituents of Dragon's blood. Part III. Dracooxepine, a novel type of biflavanoid. Heterocycles 1989, 29 (6), 1119-1125.
45. Arnone, A.; Nasini, G.; Merlini, L., Constituents of dragon's blood. Part 4. Dracoflavan A, a novel secotriflavanoid. J. Chem. Soc., Perkin Trans. 1 1990, (10), 2637-2640.
46. Nakashima, K. i.; Abe, N.; Kamiya, F.; Ito, T.; Oyama, M.; Iinuma, M., Novel Flavonoids in Dragon's Blood of Daemonorops draco. Helv. Chim. Acta 2009, 92 (10), 1999-2008.
47. Yi, T.; Tang, Y.; Zhang, J.; Zhao, Z.; Yang, Z.; Chen, H., Characterization and determination of six flavonoids in the ethnomedicine "Dragon's Blood" by UPLC-PAD-MS. Chem. Cent. J. 2012, 6, 116 (7 pages).
48. Sodi; Pallares, Arch Biochem 1946, 10, 235-241.
49. Piozzi, F.; Passannanti, S.; Paternostro, M. P.; Nasini, G., Diterpenoid resin acids of Daemonorops draco. Phytochemistry 1974, 13 (10), 2231-2233.
50. Nasini, G.; Piozzi, F., Pterocarpol and triterpenes from Daemonorops draco. Phytochemistry 1981, 20 (3), 514-516.
51. Okamoto, A.; Ozawa, T.; Imagawa, H.; Arai, Y., Flavans from the Pith of Sago Palm. Agric. Biol. Chem. 1986, 50 (6), 1655-1656.
52. Jiao, W. H.; Xu, T. T.; Yu, H. B.; Chen, G. D.; Huang, X. J.; Yang, F.; Li, Y. S.; Han, B. N.; Liu, X. Y.; Lin, H. W., Dysideanones A−C, Unusual Sesquiterpene Quinones from the South China Sea Sponge Dysidea avara. J. Nat. Prod. 2014, 77, 346-350.
53. Didierjean, C.; Marin, J.; Wenger, E.; Briand, J. P.; Aubrya, A.; Guichard, G., (6S)-6-Isobutylpiperidine-2,4-dione and (4R,6S)/(4S,6S)-4-hydroxy-6-iso-butylpiperidin-2-one. Acta Cryst. 2004, C60, o204-o207.
54. Srisanga, S.; ter Horst, J. H., Racemic Compound, Conglomerate, or Solid Solution: Phase Diagram Screening of Chiral Compounds. Crystal Growth & Design 2010, 10 (4), 1808-1812.
55. Kurkin, V. A.; Sentsov, M. F.; Zapesochnaya, G. G.; Braslavskii, V. B.; Tolkaehev, N. O., Flavonoids of the buds of Populus laurifolia. Chemistry of Natural Compounds 1994, 30 (6), 778-779.
56. Loua, H.; Yamazaki, Y.; Sasaki, T.; Uchida, M.; Tanaka, H.; Oka, S., A-type proanthocyanidins from peanut skins. Phytochemistry 1999, 51, 297-308.
57. Silva, S. M.; Abe, S. Y.; Bueno, F. G.; Lopes, N. P.; Mello, J. C. P. d.; Nakashima, T., Direct proof by 13C-nuclear magnetic resonance of semi-purified extract and isolation ofent-Catechin from leaves of Eucalyptus cinerea. Pharmacogn Mag. 2014, 10 (39), 191-194.
58. Korver, O.; Wilkins, C. K., Circular dichroism spectra of Flavanols. Tetrahedron 1971, 27, 5459-5465.
59. Xiao, X.; Si, X.; Tong, X.; Li, G., Preparation of flavonoids and diarylheptanoid from Alpinia katsumadai hayata by microwave-assisted extraction and high-speed counter-current chromatography. Separation and Purification Technology 2011, 81 (3), 265-269.
60. Li, C. W.; Cui, C. B., One new and nine known flavonoids from Choerospondias axillaries and their in vitro antitumor, anti-hypoxia and antibacterial activities. Molecules 2014, 19 (12), 21363-21377.
61. Yi, B.; Hu, L.; Mei, W.; Zhou, K.; Wang, H.; Luo, Y.; Wei, X.; Dai, H., Antioxidant phenolic compounds of cassava (Manihot esculenta) from Hainan. Molecules 2011, 16 (12), 10157-10167.
62. Monthong, W.; Pitchuanchom, S.; Nuntasaen, N.; Pompimon, W., (+)-Syringaresinol Lignan from New Species Magnolia Thailandica. Am. J. Applied Sci. 2011, 8 (12), 1268-1271.
63. Zhu, Z.; Ma, L.; Zhu, H. Y.; Yang, X. S.; Hao, X. J., Studies on the chemical constituents of Laportea bulbifera. J. Chin. Mater Med. 2011, 34 (2), 223-225.