臺灣博碩士論文加值系統

水禾(Hygroryza aristata)為禾本科(Poaceae，Gramineae)水禾屬(Hygroryza)之植物。水禾為單種屬,一年或多年生之水生草本植物,主要分部於台灣、中國南部、斯里蘭卡、印度、馬來西亞。關於本屬植物之成分和藥理活性目前尚無文獻報導,而水禾甲醇粗抽物經藥效篩選後,在抗癌、抗氣喘、心血管作用有顯著的效果,故針對其化學成分作詳細的研究。
將乾燥的水禾(948 g)以甲醇浸泡冷浸萃取,所得之濾液經減壓濃縮後,分別以水、氯仿和正丁醇進行多次分配萃取,各分配層以管柱層析分離纯化,總共分離得到35個化合物,其中29個化合物經光譜分析、文獻比對,確認其化學結構為: hygrorine-A、hygarine、N-p-coumarylanthranilic acid 、dihydroxymethyl-bis(3,5- dimethoxy-4-
hydroxyphenyl)tetrahydrofuran-9-O--xylopyranoside、rutin、quercetin、kaempferol、(-)-epigallocatechin-3-O-gallate、(-)-epicatechin-3-O-gallate、adenosine、inosine、adenine、uracil、uridine、p-coumaric acid 、methyl coumarate、 ferulic acid、n-hexacosyl-ferulate、vanillic acid、 p-hydroxybenzoic acid 、L-tyrosine、physcion、2,6-dimethoxyquinone、cycloeucalenol、-sitosterol & stigmasterol mixtures、eicosanoic acid 2,3-dihydroxypropyl ester、butcosanoic acid 2,3-dihydroxypropyl ester、sucrose、 squalane。
上述化合物中，hygrorine-A 、hygarine、dihydroxymethyl-bis(3,5-
dimethoxy-4-hydroxylphenyl)tetrahydrofuran-9-O-β-xylopyranoside為天然界首次分離得到之新化合物。其中最特別的是，hygrorine-A是從水禾植物分離得到cyclopeptide類之化合物，這是禾本科植物首次分離得到之類型。其他另有6個peptide類型之化合物尚未解出。
超氧自由基與elastase釋放測定上，dihydroxymethyl-bis(3,5-di
methoxy-4-hydroxy-phenyl)tetrahydrofuran-9-O-β-xylopyranoside具有50%以上的抑制fMLP/CB刺激嗜中性白血球所引發的超氧自由基與elastase的釋放活性，其IC50 分別為 19.33 ± 0.86 μM與24.14 ± 1.59 μM。

Hygroryza aristata is a one year or perennial aquatic herbaceous plant and widely distributed in Taiwan, southern China, Sri Lanka, India and Malaysia. The chemical and pharmacological studies of the Hygroryza aristata have not been reported previously. The methanolic crude extract of Hygroryza aristata showed significant effects for anti-tumor, anti-asthma and cardiovascular system. Therefore, we are interested in the reseach of the bioactive components of Hygroryza aristata.
The dry Hygroryza aristata (948 g) were extracted with MeOH, and then concentrated under reducing pressure to get crude extract. The methanolic extract was divided into chloroform-, n-butanol- and water- soluble layers by liquid-liquid partitioning process. These layers were chromatographed on silica gel, Diaion HP-20, Sephadex LH-20 columns, and PTLC, followed by recrystallization to give 35 compounds. The structures of 29 compounds were determined by spectral analysis and compared with literatures. They included hygrorine-A, hygarine, N-p-coumarylanthranilic acid, dihydroxymethyl-bis(3,5- dimethoxy-4-
hydroxyphenyl)tetrahydrofuran-9-O--xylopyranoside, rutin, quercetin, kaempferol, (-)-epigallocatechin-3-O-gallate, (-)-epicatechin-3-O-gallate,
adenosine, inosine, adenine, uracil, uridine, p-coumaric acid, methyl coumarate, ferulic acid, n-hexacosyl-ferulate, vanillic acid, p-hydroxybenzoic acid, L-tyrosine, physcion, 2,6-dimethoxyquinone, cycloeucalenol, β-sitosterol & stigmasterol mixtures, eicosanoic acid 2,3-dihydroxypropyl ester, butcosanoic acid 2,3-dihydroxypropyl ester, sucrose and squalane.
Among them, hygrorine-A, hygarine and dihydroxymethyl-bis(3,5-
dimethoxy-4-hydroxylphenyl)tetrahydrofuran-9-O--xylopyranoside were new compounds from natural source. Specially, hygrorine-A is a cyclopeptide and it is isolated for the first time from Gramineae family. Furthermore, dihydroxymethyl-bis(3,5-dimethoxy-4-hydroxyphenyl)-
tetrahydrofuran-9-O--xylopyranoside showed significant inhibition of superoxide anion generation and elastase release in fMLP/CB-
induced human neutrophils with IC50 values 19.33 ± 0.86 and 24.14 ± 1.59 μM, respectively.

目 錄
指導教授推薦書
口試委員審定書
國科會授權書
誌謝
目錄………………………………………………………………………i
中文摘要…………………………………………………………………ii
英文摘要………………………………………………………………...iii

第一章 緒論……………………………………………………………1
第一節 前言………………………………………………………1
第二節 水禾之植物型態與分布…………………………………1
第三節 水禾之研究目的與動機…………………………………2

第二章 水禾的萃取與分離……………………………………………5

第三章 化合物結構解析……………………………………………..17
第一節 Hygrorine-A之結構解析……..………………………...17
第二節 Hygarine之結構解析……………………………...……26
第三節 N-p-Coumarylanthranilic acid之結構解析……………..31
第四節 Dihydroxymethyl-bis(3,5-dimethoxy-4-hydroxyphenyl)
tetrahydrofuran-9-O-β-xylopyranoside之結構解析.…...34
第五節 Cycloeucalenol之結構解析.............................................40
第六節 其他化合物......................................................................45

第四章 生物活性試驗...........................................................................46
第一節 分配萃取層之生物活性..................................................46
第二節 DPPH自由基的清除作用...............................................48
第三節 超氧自由基 (O2.-)與Elastase釋放之測定…………….53
第五章 結論...........................................................................................58

第六章 實驗部分……………………………………………………..60
第一節 本實驗所用之儀器與藥品……………………………..60
第二節 抽取與分離……………………………………………..62

※光譜數據整理………………………………………………………..66
※References…..……………………..…………………………………83

1.C. C. Hsu, C. S. Kuoh, W. C. Lin, H. Y. Liu, “Flora of Taiwan” 2ed edition, Vol 5, p320-321, Epoch. Publishing Co.,Ltd.,Taiwan, 2000

2.昆明植物研究所編著,雲南植物誌第九卷種子植物, p212-214, 科學出版社, 雲南, 2003

3.鄭武燦, 台灣植物圖鑑, 下冊, p1753, 國立編譯館, 台北, 2000

4.林春吉, 台灣水生植物第一卷, p143, 田野影像出版社, 台北, 2002

5.N. H. Tan, J. Zhou, Plant cyclopeptides, Chem. Rev., 106, 840-895, 2006

6.徐任生, 天然產物化學, p201, 科學出版社, 北京, 1997

7.M. Ponchet, F. B. Jean, M. Hauteville, P. Riccl, Dianthramides(N-benzoyl and N-paracoumarylanthranilic acid derivativies) from elicited tissues of dianthus caryophyllus, Phytochemistry, 27, 2725-730, 1988

8.于德泉, 楊俊山, 分析化學手冊第七分冊核磁共振波譜分析, 第二版, p901-903, 化學工業出版社, 北京, 1999

9.T. Kanchanapoom, M. S. Kamel, R. Kasai, K. Yamasaki, C. Picheansoonthon, Y. Hiraga, Lignan glucosides from Acanthus ilicifolius, Phytochemistry, 56, 369-372, 2001

10.江一民, 正榕氣根之成份研究, 國立台灣大學博士論文, p33, 2001

11.T. M. Sarg, A. M. Ateya, N. M. Farrag, Constituents and biological activity of Biden pilosa L. grown in Egypt, Acta Pharmaceutica Hungarica, 61, 317-323, 1991

12.C. K. Lee, M. H. Chang, The chemical constituents from the heartwood of Eucalyptus citriodora, J. Chin. Chem. Soc., 47, 555-560, 2000

13.K. Kazuma, N. Noda, M. Suzuki, Malonylated flavonol, glycoside from the petals of Clitoria ternatea, Phytochemistry, 62, 229-237,2003

14.S. S. Lee, F. Y. Tsai, I. S. Chen, Chemical constituents from Berchemia formosana, J. Chin. Chem. Soc., 42, 101-105, 1995

15.R. Slimestad, M. Andersen, G. W. Francis, A. Marston, K. Hostettmann, Syringetin 3-O-(6"-acetyl)-β-glucopyranoside and other flavonols from needles of norway spruce, Picea abies, Phytochemistry, 40, 1537-1542, 1995

16.T. Kohri, F. Nanju, M. Suzuki, R. Seto, N. Matsumoto, M. Yamakawa, H. Hojo, Y. Hara, D. Desai, S. Amin, C. C. Conaway, F. L. Chung, Synthesis of (-) –[4-3H] epigallocatechin gallate and its metabolic fate in rats after intravenous administration, J. Agric. Food Chem., 49, 1042-1048, 2001

17.G. Goetz, A. Fkyerat, N. Metais, Resistance factors to grey mould in grape berries : identification of some phenolics inhibitors of Botrytis cinerea stilbene oxidase, Phytochemistry, 52, 759-767, 1999

18.W. Fan, Y. Tezuka, S. Kadota, Proly endopeptidase inhibitory activity of fourteen kampo formulas and inhibitory constituents of Tokaku-joki-to, Chem. Pharm. Bull., 48, 1055-1061, 2000

19.P. Acharya, J. Chattopadhyaya, The hydrogen bonding and hydration of 2´-OH in adenosine and adenosine 3´-ethyl phosphate, J. Org. Chem., 67, 1852-1865, 2002

20.李欣南, (1)紅花之化學成分研究 (2)核苷和核鹼基之抑制人嗜中性白血球產生超氧陰離子自由基研究, 國立國防醫學院碩士論文, p195, 1995

21.A. Takahashi, T. Nunozawa, T. Endo, S. Nozoe, Isolation of 1-β-Darabinofuranosylcytosine from the Mushroom Xerocomus nigromaculatus Hongo., Chem. Pharm. Bull., 40, 1313-1314, 1992

22.J. Kitajima, T. Tanaka, Y. Ida, Water-soluble constituents of fennel.Ⅸ. glucosides and nucleosides, Chem. Pharm. Bull., 47, 988-992, 1999

23.M. Sako, H. Kawada, A new and efficient synthetic method for 15N3-labeled cytosine nucleoside : dimroth rearrangement of cytidine N3-oxides, J. Org. Chem., 23, 8148-8150, 2004

24.Y. L. Lin, W. Y. Wang, Y. H. Kuo, C. F. Chen, Nonsteroidal constituents from Solanum incanum L., J. Chin. Chem. Soc., 47, 247-251, 2000

25.H. Koyama, T. Yoneyama, H. Fukawa, T. Shimizu, Nippon Nogei Kaishi, 43, 826, 1969

26.K. Kosuge, K. Mitsunaga, K. Koike, T. Ohmoto, Studies on the constituents of Ailanthus integrifolia, Chem. Pharm. Bull., 29, 1669-1671, 1994

27.C. C. Shen, W. J. Syu, S. Y. Li, C. H. Lin, G. H. Lee, C. M. Sun, Antimicrobial activities of naphthazarins from Arnebia euchroma, J. Nat. Prod., 65, 1857-1862, 2002

28.C. Y. Chen, F. R. Chang, C. M. Teng, Y. C. Wu, “Cheritamine, a new N-fatty acyl tryptamine and other constituents from the stems of Annona cherimola,” J. Chin. Chem. Soc., 46, 77-86, 1999

29.T. S. Wu, J. H. Yeh, P. L. Wu, The heartwood constituents of Tetradium glabrifolium, phytochemistry, 40, 121-124, 1995

30.D. R. Houck, J. L. Hanners, C. J. Unkefer, Biosynthesis of pyrroloquinoline quinine.1. Identification of biosynthetic precursors using 13C labeling and NMR spectroscopy, J. Am. Chem. Soc., 110, 6920-6921, 1988

31.E. C. Wang, M. H. Shih, M. C. Liu, M. T. Chen, G. H. Lee, Studies on constituents of Saururus chinensis, Heterocycles, 43, 969-975, 1996

32.W. Y. Lin, Y. H. Kuo, Y. L. Chang, C. M. Teng, E. C. Wang, I. Tsutomu, I. S. Chen, Anti-platelet aggregation and chemical constituents from the Rhizome of Gynura japonica, Planta Med., 69, 757-764, 2003

33.Y. C. Chang, F. Chang, Y. C. Wu, The constituents of Lindera glauca, J. Chin. Chem. Soc., 47, 373-378, 2000

34.K. Susumu, I. Toshiro, Y. Ken, Y. Sakae, T. Michio, (+)-α-Viniferin an antiflammatory compound from Caragana chamlagu root, Chem. Pharm. Bull., 38, 432-435, 1990

35.A. M. Wolters, D. A. Jayawickrama, C. K. Larive, J. V. Sweedler, Capillary isotachophoresis/NMR : extension to trace impurity analysis and improved instrumental coupling, Anal. Chem. 74, 2306-2313, 2002

36.T. S. Wu, F. C. Chang, P. L. Wu, C. S. Kaoh, I. S. Chen, Constituents of Tetradium glabrifolium, J. Chin. Chem. Soc., 42, 929-934, 1995

37.A. Mellors, A. L. Tappel, The inhibition of mitochondrial peroxidation by ubiquinone and ubiquinol, J. Biol. Chem., 241, 4353-4356, 1966

38.C. Dahlgren, A. Karlsson, Respiratory burst in human neutrophils, J. Immunol. Methods, 232, 3-14, 1997