Terpenes from the Aerial Parts of Chrysanthemum coronarium L.#

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(1)Note. Agric. Chem. Biotechnol. 46(3), 118-121 (2003). Terpenes from the Aerial Parts of Chrysanthemum coronarium L.. #. Myoung-Chong Song, Dong-Hyun Kim, Yoon-Hee Hong, Dae-Keun Kim1, In-Sik Chung, Sung-Hoon Kim2, Mi-Hyun Park3, Byoung-Mog Kwon4, Youn-Hyung Lee* and Nam-In Baek Graduate School of Biotechnology & Plant Metabolism Research Center, KyungHee University, Suwon 449-701, Korea 1 Department of Pharmacy, Woosuk University, Jeunbuk 565-701, Korea 2 Graduate School of East-West Medical Science, KyungHee University, Suwon 449-701, Korea 3 Erom Life Co. Ltd., Seoul 135-825, Korea 4 Korea Research Institute of Bioscience and Biotechnology, KIST, Taejon 305-333, Korea Received August 28, 2003; Accepted September 17, 2003. The aerial parts of Chrysanthemum coronarium L. were extracted with MeOH, and the extract was partitioned using EtOAc, n-BuOH, and H2O. The repeated column chromatography of the EtOAc and n-BuOH fractions gave two terpenes, phytol and 8-hydroxylinalol 8-O-β-D-glucopyranoside, identified by several spectral analyses, including NMR and MS. This paper is the first report on the isolation of these terpenes from Chrysanthemum coronarium L. Key words : Chrysanthemum coronarium, phytol, 8-hydroxylinalol 8-O-β-D-glucopyranoside.. During the course of evaluating edible plants for biological activity, the authors reported the isolation of secondary metabolites with biological functionality from several plants such as sweet potatoes,1) pumpkin leaves,2) a wild garlic,3,4) and lettuce.5) Chrysanthemum coronarium L. (Compositae) is an annual herb, which has been favorably ingested in the Korean diet because of its fragrant flavor as well as abundance in nutritional value. Moreover, the aerial parts of C. coronarium have been used for the protection or remedy of several diseases in oriental medicinal systems.6) Several constituents have been isolated from C. coronarium. These include essential oils,7) quercetin, quercetagetin, luteolin,8) polyacetylenes,9-11) chrycorin and chrycolide,11) sesquiterpenes lactones, cumambrin A and dihydrocumambrin A,12) douglanine and reynosin,13) dihydrochrysanolide, 1α-epidihydrochrysanolide and 1-hydroxy-1-desoxotamirin,14) 8αangeloyloxy-10α-hydroxyslov-3-en-6,12-olide and tigloylcumambrin B,15) dihydrotulipinolide,16) pyrethrosin, 1,10-epi-pyrethrosin, and tulirinol17). However, the relationship between biological activity and the constituents still has not been reported. This paper examines the isolation of two terpenes, which were first isolated from this herb, and structure determination using spectral data.. #. This paper is Part VII of Search for Biologically-Active Materials from Edible Plant Resources. *Corresponding author Phone: 82-31-201-2670; Fax: 82-31-201-2157 E-mail: [email protected]. Materials and Methods Instrumentation. Optical rotations were measured on a JASCO DIP-370 digital polarimeter. EI mass spectra were taken on a JEOL JMS-AX505WA spectrometer. IR spectra were run on a Perkin Elmer Spectrum One FT-IR spectrometer. The 1H- and 13C-NMR spectra were taken on a Varian Unity Inova AS 400 FT-NMR spectrometer. Plant materials. C. coronarium L. was purchased from a farm located in Yangju-Si, Korea in December, 2002. A voucher specimen (KHU020809) was reserved at the Laboratory of Natural Products Chemistry, KyungHee University, Suwon, Korea. Isolation of terpenes. The fresh aerial parts of C. coronarium (80 kg) were extracted at room temperature with MeOH (40 L × 3) for 24 hr. The filtrate was concentrated in vacuo at 40oC to render the MeOH extracts. The extracts were successively partitioned with water (2 L), EtOAc (2 L × 3), and n-BuOH (2 L × 3) to yield the EtOAc (168 g), n-BuOH (126 g), and water (512 g) extracts. The EtOAc extract (168 g) was applied to silica gel (1500 g) column chromatography (nhexane-EtOAc = 7 : 1→ 6 : 1→ 5 : 1→ 4 : 1→ 3 : 1→ 2 : 1 →1 : 1) monitoring by thin layer chromatography (TLC) to produce twelve fractions (CSE1~CSE12). The fifth fraction (CSE5, 13.3 g) was applied to the silica gel (260 g) column chromatography (n-hexane-EtOAc = 8 : 1) to yield thirteen fractions (CSE51~CSE513). The fourth fraction (CSE54, 4.3 g) was subjected to the silica gel (200 g) column chromatography (n-hexane-EtOAc-MeOH = 10 : 1 : 1) and afforded eleven fractions (CSE541~CSE5411). The sixth fraction (CSE546, 1.2 g) was applied to the silica gel (100 g) column chromatography (n-hexane-EtOAc = 8 : 1) to yield.

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(31) . Results and Discussion Fig. 1. Chemical structure of the terpene compounds from the aerial parts of Chrysanthemum coronarium L. (Garland).. eleven fractions (CSE5461~CSE54611). The fifth fraction (CSE5465, 256 mg) compound 1 (200 mg) was isolated using silica gel (100 g) column chromatography (n-hexane-EtOAc = 8 : 1). &RPSRXQG SK\WRO

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(64) The n-BuOH extract (92 g) was applied to silica gel (800 g) column chromatography (CHCl3-MeOH = 10 : 1→9 : 1→8 : 1→7 : 1→6 : 1) to produce twelve fractions (CSB1~CSB12). The sixth fraction (CSB6, 9.0 g) was applied to the silica gel (200 g) column chromatography (CHCl3-MeOH-H2O = 7 : 3 : 1) to yield eight fractions (CSB61~CSE68). The fifth one (CSE65, 572 mg) was subjected to the silica gel (50 g) column chromatography (CHCl3-MeOH-H2O = 9 : 3 : 1) to afford six fractions (CSB651~CSB656). Then the fourth fraction (CSB654, 300 mg) was chromatographed on silica gel (90 g) column (CHCl3-MeOH-H2O = 9 : 3 : 1) to yield six fractions (CSB6541~CSB6546). The final isolation of compound 2 (CSB65449, 11 mg) was carried out using ODS (25 g) column chromatography (MeOH-H2O = 4 : 1) of the fourth fraction (CSB6544, 101 mg). &RPSRXQG K\GUR[\OLQDORO2β'JOXFRS\UDQRVLGH

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(77) + G. To search biologically-active materials from C. coronarium, the aerial parts of the plant were extracted with MeOH. The extracts were sequentially partitioned with EtOAc, n-BuOH, and water. Repeated silical gel or ODS column chromatography of the EtOAc and n-BuOH fractions led to the isolation of two terpenes-compounds 1 (yield; 2.0 × 10−6 %) and 2 (yield; 1.5 × 10−7 %), respectively. Compound 1, colorless oil (n-hexane-CHCl3), [α]D +0.2o, had hydroxyl and double bond from the bands of 3334 and 1699 cm−1, respectively, in the IR spectrum (KBr). Also, the comparison of NMR and MS data with those in literature18,19) led to the identification of compound 1 as an acyclic diterpene alcohol, phytol, possessing a R configuration at two asymmetric carbons (C7 and C11) and an E configuration of the double bond.20,21) Phytol occurs abundantly in plants as an alcohol part of the ester sidechain of chlorophyll. It is an useful material for the syntheses of α-tocopherol (vitamin E)22) and phylloquinone (vitamin K1). In the IR spectrum compound 2, colorless oil (MeOH), [α]D +11o, contained hydroxyl, double bond, and exomethylene from the bands of 3580, 1605, and 925 cm−1, respectively. In the 1HNMR spectrum (400 MHz), two olefine proton signals of two olefine methine [δH 6.10 (dd, J = 17.6, 10.8 Hz), 5.64 (1H, t, J = 6.8 Hz)] and two olefine ones of exomethylene [δH 5.51 (dd, J = 17.6, 2.0 Hz), δH 5.11 (dd, J = 10.8, 2.0 Hz)] were observed. A hemiacetal signal at δH 4.85 (d, J = 8.0 Hz) and several oxygenated methine and methylene signals that were observed at δH 3.91-4.52 indicated the presence of one sugar in the molecule. At the high magnet field, two methylene proton signals [δH 2.30 (2H, m), δH 1.70 (2H, m)] and two methyl ones [δH 1.72 (3H, s), δH 1.41 (3H, br. s)] were observed. In the 13CNMR spectrum (100 MHz), sixteen carbon signals {including those of one D-glucopyranose that was identified from the chemical shifts of one hemiacetal (δC 103.41), four oxygenated methines (δC 78.47, 78.47, 75.18, 71.67), and one oxygenated methylene (δC 62.28) signals} were observed indicating that compound 2 was a monoterpene-glucoside. Also, one olefine quaternary (δC 132.12), two olefine methine (δC 146.78, 128.76), and one exomethylene (δC 111.30) signal (derived from two double bonds, along with one oxygenated quaternary (δC 72.29) and one oxygenated methylene (δC 75.06) signals) were observed. At the high magnet field, two methylene (δC.

(78) 120. Myoung-Chong Song et al.. 42.82, 28.48) and two methyl (δC 23.06, 14.38) signals were also observed. Finally, the aglycone of compound 1 was determined as 8-hydroxylinalol23) from these results and the 1H1 H COSY experiment. The glycosidation shift of C-8 oxygenated methylene indicated that the glucose was linked at the hydroxy of C-8. This was confirmed by the gHMBC spectrum, in which the oxygenated methylene at δC 75.06 (C-8) showed a correlation with the anomeric proton at δH 4.85, the methyl proton at H 1.41 (H-10), and the olefine proton at δH 5.64 (H-6). The configuration of the glucopyranose was determined as β from the coupling constant (8.0 Hz) of the anomeric proton signal in the 1H-NMR spectrum of compound 2. Therefore, compound 2 was identified as 3,7-dimethyl-3,8-dihydroxy-oct1,6-diene 8-O-β-D-glucopyranoside (8-hydroxylinalol 8-O-βD-glucopyranoside).23) Presently, the isolated two terpene compounds have not been isolated from this plant, C. coronarium. Acknowledgements. This study was supported by the BioGreen 21 Program from the Rural Development Administration, Republic of Korea, and by a grant from the Korea Science & Technology Foundation through the Plant Metabolism Research Center, KyungHee University.. References 1. Baek, N.-I., Ahn, E. M., Bang, M. H. and Kim, H. Y. (1997) Development of biologically active compounds from edible plant sources-I. Isolation of major components from the tuber of Ipomoea batatas Lam. J. Korean Soc. Agric. Chem. Biotechnol. 40, 583-587. 2. Han, J. T., Ahn, E. M. and Baek, N.-I. (1999) Development of biologically active compounds from edible plant sources-II. Isolation of fatty acids and sterol glycosides from the leaves of Cucurbita moschata DUCH. Ibid. 42, 267-270. 3. Ahn, E. M., Jang, T. H. and Baek, N.-I. (2000) Development of biologically active compounds from edible plant sources-III. Isolation of flavonoid-glycosides from the Allium monanthum Max. Ibid. 43, 314-316. 4. Baek, N.-I., Ahn, E. 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(79) &KHPLVWU\ RI YLWDPLQ ( ,,, 7RWDO V\QWKHVLV RI 5 5 5

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(81) αWRFRSKHURO,ELG 22. Burrell, J. W. K., Garwood, R. F., Jackman, L. M., Osaky, E. and Weedon B. C. L. (1966) Carotenoids and related compounds. XIV. Stereochemistry and synthesis of geran-.

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