Chinese Journal of Natural Medicines 2013, 11(5): 0534−0537

Chinese Journal of Natural Medicines

A new megastigmane glycoside from the aerial parts of Cirsium setosum JIANG Hai, MENG Yong-Hai, YANG Liu, WANG Qiu-Hong, YANG Bing-You, LIU Chang, KUANG Hai-Xue* Key Laboratory of Chinese Materia Medica (Heilongjiang University of Chinese Medicine), Ministry of Education, Harbin 150040, China Available online 20 Sept. 2013

[ABSTRACT] AIM: To study the chemical constituents of the aerial parts of Cirsium setosum (Willd.) MB.. METHODS:The chemical constituents were isolated and purified by various chromatographic techniques. Their structures were determined on the basis of physical properties and spectroscopic data. RESULTS: A new megastigmane glycoside and six known compounds were obtained and identified as (7E, 9R)-9-hydroxy-5, 7-megastigmadien-4-one-9-O-α-L-arabinopyranosyl-(1→6)-β-D-glucopyanoside (1), (6R, 7E, 9R)-9-hydroxy-4, 7-megastigmadien-3-one-9-O-α-L-arabinopyranosyl-(1→6)-β-D-glucopyranoside (2), urolignoside (3), 4, 9, 9'-trihydroxy-3, 3'-dimethoxy-8-O-4′-neolignan-7-O-β-D-glucopyranoside (4), citroside A (5), salidrosidin (6), and adenosine (7). CONCLUSION: Compound 1 is a new megastigmane glycoside, named as Xiaojiglycoside A.

[KEY WORDS] Compositae; Cirsium setosum; Megastigmane glycoside; Xiaojiglycoside A [CLC Number] R284.1 [Document code] A [Article ID] 1672-3651(2013)05-0534-04



Cirsium setosum (Willd.) MB.(Compositae) is a perennial herb. It is a famous traditional Chinese medicine (TCM) called “Xiao-Ji” and has been used for thousands of years to treat hematuria, spitting of blood, and uterine bleeding. Recent pharmacological studies of this plant have shown a variety of activities including anti-inflammatory [1], antimicrobial [2] , antihemorrhagic [3], anticancer [4], sedative [5]. Previous phytochemical investigations led to the isolation of flavonoids, sterols, organic acids, phenolic acids, alkaloids, triterpenes [6-10], and the flavonoids were found to be the active components for the antihemostatic, anti-inflammation, antitumor, sedative, analgesic, and antipyretic [8, 11-15]. During our systematic investigation on its chemical constituents, six known compounds were isolated. Compound 1 is a new megastigmane glycoside isolated from C. setosum. Its structure was elucidated on the basis of the spectroscopic [Received on] 11-Apr.-2012 [Research funding] This project was supported by the Administration of Traditional Chinese Medicine of Heilongjiang Province project (ZHY06-Z05). [*Corresponding author] KUANG Hai-Xue: Prof., Tel: 86-45182193001, Fax: 86-451-82110803, E-mail:[email protected] These authors have no any conflict of interest to declare. Published by Elsevier B.V. All rights reserved

analysis, including 2D-NMR techniques, HR-ESI-MS and the result of hydrolytic reaction. Herein, we report the isolation and its structural elucidation.


Results and Discussion Compound 1

mp 156−157 °C, [α] 20 −18.3 (c 0.18, D

MeOH), produced positive in the Molish reaction, HR-ESIMS ([M + H]+ 503.249 0, calcd. for C24H39O11+ 503.249 2). IR (KBr) vmax: 3 450, 2 890, 1 653, 1 450, 1 070, 1 042 cm−1. UV λmax (MeOH) nm (logε): 203 (3.16), 260 (3.50). 1H and 13 C NMR data, see Table 1. Compound 1 was obtained as white amorphous powder. The 1H NMR spectrum showed the presence of three singlet and one doublet methyls, two trans-olefinic signals at δ 6.32 (1H, d, J = 16.4 Hz, H-7) and 5.76 (1H, dd, J = 16.4, 6.4 Hz, H-8) and two anomeric proton signals of sugar moieties at δ 4.38 (1H, d, J = 7.6 Hz, H-1′) and 4.27 (1H, d, J = 6.4 Hz, H-1′′). The 13C NMR spectrum of 1 indicated the presence of two sugar units, and the remaining 13 signals were expected to present a megastigmane skeleton [16]. These signals were assigned as four methyls, two methylenes, three methines, four quaternary carbon (including, one carbonyl carbon and two olefinic carbons). The 1H-1H COSY spectrum revealed the correlations between H2-2 and H2-3, and the correlations between H-7, H-8, H-9 and H3-10. In the HMBC spectrum (Fig. 1), some key long-range correlations were observed

JIANG Hai, et al. /Chinese Journal of Natural Medicines 2013, 11(5): 534−537

between H2-2 and C-1, C-3, C-4 and C-6, as well as H2-3 and C-2, C-4 and C-5, and between H3-10 and C-7 and C-9, as well as H3-13 and C-4, C-5 and C-6, respectively. Table 1 1H NMR and 13C NMR data for 1 (400 and 100 MHz, CD3OD, J in Hz) No 1 2 3 4 5 6 7 8 9 10 11 12 13 1′ 2′ 3′ 4′ 5′ 6′ 1′′ 2′′ 3′′ 4′′ 5′′

δH 1.85 (2H, t, J = 6.4 Hz) 2.46 (2H, t, J = 6.4 Hz)

6.32 (1H, d, J = 16.4 Hz) 5.76 (1H, dd, J = 16.4, 6.4 Hz) 4.47 (1H, dq, J = 6.4, 6.2 Hz) 1.34 (3H, d, J = 6.2 Hz) 1.17 (3H, s) 1.18 (3H, s) 1.78 (3H, s) 4.38 (1H, d, J = 7.6 Hz) 3.20 (1H, t, J = 8.4 Hz) 3.34 (1H, t, J = 8.4 Hz) 3.34 (1H, m) 3.40 (1H, m) 3.68 (1H, dd, J = 12, 5.2 Hz) 3.75 (1H, dd, J = 10.8, 2.8 Hz) 4.27 (1H, d, J = 6.4 Hz) 3.59 (1H, m) 3.37 (1H, m) 3.69 (1H, br.s) 3.57 (1H, m) 3.80 (1H ,dd, J = 12.0, 3.2 Hz)

δC 36.6 38.2 35.1 201.8 127.5 163.7 130.7 140.3 77.2 20.9 27.7 27.7 13.7 102.7 75.2 77.9 71.4 77.2 69.2 105.0 72.3 74.1 69.4 66.6

Fig. 1 Structure and key HMBC and 1H-1H COSY correlations of compound 1

The above information indicated a 9-hydroxy-5, 7-megastigmadien-4-one skeleton [16]. The acid hydrolysis of 1 liberated D-glucose and L-arabinose, which were identified by HPLC analysis using an optical rotation detector [17]. Two anomeric proton signals were assigned to two anomeric carbon signals at δ 102.7 (C-1′), 105.0 (C-1′′) in the HMQC experiment, respectively. By comparing coupling constants and chemical shift of signals of sugar moieties with those reported [18-20], the glucose and the arabinose moieties were deduced to be β and α configuration respectively. Moreover, the connection positions of the sugar groups were established unambiguously by the HMBC correlations between H-1′ and

C-9, and H-1′′ and C-6′. Comparison of 13C NMR chemical shifts in 1 with those of platanionoside F [16] indicated the presence of 9 R configuration. On the basis of the above analysis, compound 1 was elucidated as (7E, 9R)-9-hydroxy-5, 7-megastigmadien-4-one-9O-α-L-arabinopyranosyl-(1→6)-β-D-glucopyranoside. It was a new compound and named as Xiaojiglycoside A. By comparison of 1H and 13C NMR data and physical data with those reported in the literatures compounds 2-7 were identified as (6R, 7E, 9R)-9-hydroxy-4, 7-megastigmadien-3-one-9-O-α-L-arabino-pyranosyl-(1→6)β-D-glucopyranoside (2) [21], urolignoside (3) [22] 4, 9, 9'-trihydroxy-3, 3'-dimethoxy-8-O-4′neolignan-7-O-β-D-glucopyranoside (4) [23], citroside A (5) [24], salidrosidin (6) [25], and adenosine (7) [26]. Compounds 2−7 were isolated from the plant species for the first time.


Materials and Methods

3.1 Equipments The melting point was measured on Kofler micromelting point apparatus (uncorrected). The optical rotation was recorded on a Perkin-Elmer 241. optical-rotation detector (Shodex OR-2, Showa Denko Co., Ltd., Japan). IR spectra were recorded on an IR-47 spectrometer. The UV and NMR spectra were recorded on SHIMADZU UV-1601 and Bruker DPX 400 (400 MHz for 1H NMR and 100 MHz for 13C NMR), respectively. Chemical shifts are given as δ values with reference to tetramethylsilane (TMS) as an internal standard, and coupling constants are given in Hz. The HR-ESI-MS analyses were conducted on IonSpec Ultima 7.0T FTICR. Preparative HPLC (Waters, Delta 600-2487) was performed on Hypersil-ODS Ⅱcolumn (10 μm, 20 mm × 300 mm, Yilite, Dalian, China). Silica gel (200−300 mesh, Yanghai, Qingdao, China) and ODS-A (120 A, 50 µm, YMC Co.) were employed for column chromatography (CC). 3.2 Plant material The aerial parts of C. setosum (Willd.) MB. were collected from Taoshan Forestry Bureau of Heilongjiang Province, China in 2006 and identified by Prof. WANG Zhen-Yue of Heilongjiang University of Chinese Medicine. The voucher specimen (No. 2006079) was deposited at the Herbarium of Heilongjiang University of Chinese Medicine, Harbin, China. 3.3 Extraction and isolation The air dried aerial parts of C. setosum (10 kg) were extracted with 95% EtOH under reflux (3 × 30 L) for each 2 h (each). The 95% EtOH extract (450 g) was suspended in H2O and partitioned successively with petroleum ether (3 × 2 L), CH2Cl2 (3 × 2.5 L), EtOAc (3 × 2.5 L) and n-BuOH (3 × 2.5 L). The n-BuOH fraction (80 g) was applied to a silica gel CC using a gradient of CH2Cl2−MeOH (15 : 1 to 1 : 1) to give fractions Fr1−Fr7. Fr2 (26 g) was separated by silica gel CC using a gradient of CH2Cl2−MeOH (10 : 1 to 5 : 1), to afford a number of sub-fractions A1−A5. Compound 5 (25

JIANG Hai, et al. /Chinese Journal of Natural Medicines 2013, 11(5): 534−537

mg) was obtained by ODS CC of the sub-fraction A1 (2 g) eluted with MeOH/H2O (1 : 4). Similarly, compounds 6 (21 mg) and 7 (17 mg) were obtained by ODS CC of the sub-fraction A2 (5 g) using a gradient of MeOH−H2O (1 : 4 to 1 : 0). A3 (2.8 g) was subjected to CC on ODS using a gradient of MeOH−H2O (1 : 4 to 1 : 0) to afford a number of sub-fractions B1−B4. B2 (1.6 g) was purified by preparative HPLC on a Hypersil-ODS Ⅱ column (10 μm, 20 mm × 300 mm, flow rate 8 mL·min−1) with MeOH−H2O (3 : 7) to afford compounds 1 (33 mg, tR = 19.8 min) and 2 (35 mg, tR = 21.5 min). The sub-fraction A4 (3.2 g) was also passed over ODS CC eluted with MeOH−H2O (3 : 7), and finally purified by preparative HPLC on a Hypersil-ODS Ⅱ column (10 μm,



[10] [11]



20 mm × 300 mm, flow rate 8 mL·min ) eluted with MeOH− H2O (2 : 5) to afford compounds 3 (36 mg, tR = 27 min) and 4 (20 mg, tR = 32.5 min). 3.4 Acid hydrolysis Acid hydrolysis of 1 One mL of HCl (1 mol·L−1) was added into a solution of 1 (15 mg) in MeOH (5 mL) and the mixture was refluxed for 3 h. The reaction mixture was then neutralized with saturated sodium carbonate and extracted with ethyl acetate (EtOAc, 2 × 10 mL) to give an aqueous fraction containing sugars and EtOAc fraction containing the aglycone part. The aqueous layer was evaporated and then subjected to HPLC analysis using Hypersil NH2 (5 μm, 4.0 mm × 200 mm, Yilite, Dalian, China). D-Glucose and L-arabinose were confirmed by comparison of the retention times with the authentic samples. Mobile phase: CH3CN−H2O (75 : 25), flow rate: 0.5 mL·min−1, +25.5), tR = 13.1 min. tR = 14.0 min (D-glucose, [α] 20 D






(L-arabinose, [α]20 +27.3). D

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小蓟中一个新的大柱香波龙烷苷 姜

海, 孟永海, 杨

柳, 王秋红, 杨炳友, 刘

畅, 匡海学*

黑龙江中医药大学省部共建重点实验室, 哈尔滨 150040 【摘



龙烷苷和六个已知化合物被分离得到,分别是(7E, 9R)-9-hydroxy -5, 7-megastigmadien-4-one-9-O-α-L-arabinopyranosyl-(1→6)-β-Dglucopyanoside (1), (6R, 7E, 9R)-9-hydroxy-4, 7-megastigmadien-3-one-9-O-α-L-arabinopyranosyl-(1→6)-β-D-glucopyranoside (2), urolignoside (3), 4, 9, 9'-trihydroxy-3, 3'-dimethoxy-8-O-4′-neolignan-7-O-β-D-glucopyranoside (4), citroside A (5), 红景天苷 (6), 和腺 苷 (7)。结论:化合物 1 为新化合物。 【关键词】


菊科;小蓟;大柱香波龙烷苷;小蓟苷 A 黑龙江省中医药管理局项目(No. ZHY06-Z05)资助

A new megastigmane glycoside from the aerial parts of Cirsium setosum.

To study the chemical constituents of the aerial parts of Cirsium setosum (Willd.) MB...
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