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Saponins from Lonicera bournei
Phytochemistry 54 (2000) 795±799
www.elsevier.com/locate/phytochem
Saponins from Lonicera bournei Ting Xiang a, Yasuhiro Tezuka b, Li-Jun Wu a,*, Arjun H. Banskota b, Shigetoshi Kadota b a
Department of Natural Medicines, Shenyang Pharmaceutical University, Shenyang 110015, People's Republic of China b Institute of Natural Medicine, Toyama Medical and Pharmaceutical University, Toyama, Japan Received 13 October 1999; received in revised form 28 February 2000
Abstract The lupane-triterpene glycosides, bourneioside A and bourneioside B, and two known saponins were isolated from Lonicera bournei Hemsl. The structures of bourneioside A and B were elucidated as 3-O-b-D-glucopyranosyl-23-hydroxy-lup-20(29)-en-28oic acid-28-O-b-D-glucopyranosyl ester and 3-O-b-D-glucopyranosyl-23-hydroxy-lup-20(29)-en-28-oic acid-28-O-[b-Dglucopyranosyl-(1 4 6)-b-D-glucopyranosyl] ester, respectively, on the basis of spectral data and chemical evidence. 7 2000 Published by Elsevier Science Ltd. Keywords: Lonicera bournei Hemsl; Caprifoliaceae; Bourneioside A; Bourneioside B
1. Introduction
2. Results and discussion
The ¯ower buds of Lonicera bournei Hemsl. and L. japonica Thunb. are widely used in northwest and southwest China. Both are used as antibacterial, antiin¯ammatory agents, as well as folk medicines to treat encephalitis, in¯uenza, fever, pneumonia, dysentery, enteritis and so on. Several tannins and triterpenes from L. japonica Thunb. showed inhibitory eect on HIV-1 RT and a cytoprotective eect against CCl4induced hepatic injury, respectively (Chang et al., 1995; Lou et al., 1995). This paper reports the isolation and structure elucidation of two new lupane-triterpene glycosides named bourneioside A (1) and bourneioside B (2) along with two known saponins (3 and 4) from L. bournei Hemsl. No correlation with these compounds to any medicinal property was made in this study.
The ¯ower buds of L. bournei Hemsl. were extracted with 90% ethanol. The extract was further isolated as described in Section 3 to provide two new lupane-triterpene glycosides 1 (15.7 mg) and 2 (12.6 mg) (see Fig. 1) together with two known triterpenoid glycosides, 3-O-b-D-glucopyranosyl-hederagenin-28-O-b-Dglucopyranosyl ester 3 (8.0 mg) and 3-O-[b-D-glucopyranosyl-(1 4 2)-b-D-glucopyranosyl]-hederagenin-28-O[b-D-glucopyranosyl(1 4 6)-b-D-glucopyranosyl]ester 4 (12.0 mg). Bourneioside A (1) was obtained as white crystals (MeOH) with mp 205±2078C, having aD21 +55.08 (MeOH, c 0:1), and it responded positively to the Liebermann-Burchard and Molisch tests. HR-FABMS gave a M Na]+ peak at m/z 819.4515 corresponding to the molecular formula of C42H68O14. The IR spectrum showed the presence of hydroxyl (3400 cmÿ1), carboxyl (1741 cmÿ1) and exo-methylene (1639, 890 cmÿ1) groups. The 13C NMR and DEPT spectra of the aglycone moiety exhibited 30 carbon signals (seven quaternary, six methine, 12 methylene and ®ve methyl carbons). The presence of a methyl group attached to a terminal double bond was con®rmed by the 1H
* Corresponding author. Department of Natural Medicines, Faculty of Traditional Chinese Medicine, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang, 110015, People's Republic of China. E-mail address: [email protected] (T. Xiang).
0031-9422/00/$ - see front matter 7 2000 Published by Elsevier Science Ltd. PII: S 0 0 3 1 - 9 4 2 2 ( 0 0 ) 0 0 1 9 4 - 1
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NMR signals at d 4.86 (1H, br.s ), 4.71 (1H, br.s ), 1.72 (3H, s ) and the 13C NMR signals at d 150.8, 110.1, 19.4. One primary hydroxyl group was assigned to the C-23 position based on the presence of hydroxy methylene signals at d 4.33 (1H, m ), 3.69 (1H, d, J 10:8 Hz) and d 64.6. The carbonyl carbon signal observed at d 174.9 was assigned to C-28. The above data suggested that 1 was a derivative of 23-hydroxylupanetriterpene (Nakatani et al., 1989; Ikuta and Itokawa, 1988; Ye et al., 1996). Acid hydrolysis conducted on TLC yielded only glucose. The anomeric carbon signals at d 105.8 and 95.4 indicated two glucosyl units within 1 that were con®rmed as b-D-glucosyl units by the two anomeric proton signals at d 5.12 (1H, d, J 7:8 Hz) and 6.42 (1H, d, J 8:0 Hz). The TOCSY spectrum demonstrated that the proton signals at d 5.12, 4.50 (dd, J 11:7, 2.2 Hz), 4.35 (m ), 4.22 (m ), 4.14 (m ), 4.02 (m ), 3.87 (m ) and d 6.42, 4.45 (dd, J 12:4, 2.4 Hz), 4.39 (m ), 4.35 (m ), 4.28 (m ), 4.17 (m ), 4.03 (m ) were due to two dierent glucoses, respectively. In the 1H±1H COSY spectrum, correlation peaks could be observed from the following pairs: d 5.12 (H-1')/4.02 (H-2 '), 4.02 (H2 ')/4.14 (H-3'), 4.14 (H-3 ')/4.22 (H-4'), 4.22 (H-4 ')/ 3.87 (H-5 '), 3.87 (H-5 ')/4.50, 4.35 (H-6 '); d 6.42 (H10)/4.17 (H-20), 4.17 (H-20)/4.28 (H-30), 4.28 (H-30)/ 4.35 (H-40), 4.35 (H-40)/4.03 (H-50), 4.03 (H-50)/4.45, 4.39 (H-60). Thus, the proton signals of the sugar moieties were assigned one by one. Furthermore, the carbon signals of the sugar moieties were identi®ed by means of HMQC-fg spectrum. As shown in Fig. 2, the long-range correlations between d 5.12 (H-1 ') and 82.0 (C-3), as well as d 6.42
Fig. 1. Structures of bourneioside A (1) and bourneioside B (2) from L. bournei Hemsl.
(H-10) and 174.9 (C-28) suggested that the two glucosyl units were attached to C-3 and C-28 of the aglycone, respectively. Therefore, compound 1 was established as 3-O-b-D-glucopyranosyl-23-hydroxy-lup20(29)-en-28-oic acid-28-O-b-D-glucopyranosyl ester, named bourneioside A. Bourneioside B (2) was obtained as white crystals (MeOH) with mp 214±2168C, having aD21 +40.08 (MeOH, c 0:1), and it also responded positively to the Liebermann-Burchard and Molisch tests. HRFABMS showed M Na]+ peak at m/z 981.5036 corresponding to molecular formula of C48H78O19. The 1 H NMR and 13C NMR spectra of the aglycone moiety of 2 were similar to those of 1, which indicated that 2 was also a 23-hydroxylupane-triterpene derivative. Acid hydrolysis carried out on TLC yielded only glucose. The anomeric carbon signals at d 105.7, 105.4 and 95.2 indicated three glucosyl units within 2 that were con®rmed as b-D-glucosyl units by the three corresponding anomeric proton signals at d 5.10 (1H, d, J 8:0 Hz), 6.33 (1H, d, J 8:0 Hz), 4.99 (1H, d, J 7:6 Hz). The TOCSY spectrum revealed that the proton signals at d 5.10, 4.48 (m ), 4.36 (m ), 4.18 (m ), 4.12 (m ), 4.01 (m ), 3.86 (m ); d 6.33, 4.69 (d, J 10:7 Hz), 4.32 (m ), 4.28 (m ), 4.16 (m ), 4.10 (m ), 4.07 (m ) and d 4.99, 4.46 (m ), 4.34 (m ), 4.20 (m ), 4.16 (m ), 3.97 (m ), 3.86 (m ) were due to three dierent glucoses, respectively. In the 1Hÿ1H COSY spectrum, the correlation peaks could be observed from the following pairs: d 5.10 (H-1 ')/4.01 (H-2 '), 4.01 (H-2 ')/4.12 (H3 '), 4.12 (H-3 ')/4.18 (H-4'), 4.18 (H-4 ')/3.86 (H-5 '), 3.86 (H-5 ')/4.48, 4.36 (H-6'); d 6.33 (H-10)/4.07 (H-20), 4.07 (H-20)/4.16 (H-30), 4.16 (H-30)/4.28 (H-40), 4.28 (H-40)/4.10 (H-50), 4.10 (H-50)/4.69, 4.32 (H-60); d 4.99 (H-11)/3.97 (H-21), 3.97 (H-21)/4.16 (H-31), 4.16 (H31)/4.20 (H-41), 4.20 (H-41)/3.86 (H-51), 3.86 (H-51)/ 4.46, 4.34 (H-61). These analyses clari®ed the assignments of the proton signals of sugar moieties. The carbon signals of the sugar moieties were identi®ed by means of HMQC-fg spectrum. The anomeric proton signals at d 6.33 (H-10), 5.10 (H-1'), 4.99 (H-11) showed cross-peaks with the carbon signals at d 174.9 (C-28), 82.0 (C-3), 69.5 (C-60) in the HMBC-fg spectrum, respectively. The carbon signal at d 69.5 showed correlation with proton signals at d 4.69 (1H, d, J 10:7 Hz) and 4.32 (1H, m ) which were determined to be H-60 of the glucosyl moiety attached to C-28 by TOCSY spectrum. Thus, compound 2 was established as a new 23-hydroxylupanetriterpene saponin, 3-O-b-D-glucopyranosyl-23hydroxy-lup-20(29)-en-28-oic acid-28-O-[b-D-glucopyranosyl-(1 4 6)-b-D-glucopyranosyl] ester, named bourneioside B. Compound 3 was obtained as white crystals (MeOH) with mp 211±2138C, and it responded
T. Xiang et al. / Phytochemistry 54 (2000) 795±799
positively to the Liebermann-Burchard and Molisch tests. Acid hydrolysis conducted on TLC yielded only glucose. The 1H NMR and the 13C NMR spectral data revealed that the aglycone moiety of 3 was hedergenin (Takemoto et al., 1984). The anomeric signals at d 105.8 and 95.7 indicated two glucosyl units within 3 that were con®rmed as b-D-glucosyl units by the two anomeric proton signals at d 5.11 (1H, d, J 7:8 Hz), 6.32 (1H, d, J 7:8 Hz). Thus, 3 was elucidated as 3-O-b-D-glucopyranosyl-hederagenin-28-O-bD-glucopyranosyl ester by comparison with literature and it was ®rst isolated from Lua cylindrica Roem. (Takemoto et al., 1984). Compound 4 was obtained as white crystals (MeOH) with mp 146±1488C, and it responded positively to the Liebermann-Burchard and Molisch tests. Acid hydrolysis conducted on TLC yielded only glucose. The 1H NMR and the 13C NMR spectral data of the aglycone moiety of 4 were similar to those of 3, which indicated that the aglycone moiety of 4 was also hedergenin. The sugar moieties of 4 were dierent from those of 3. The anomeric signals at d 105.9, 105.3, 103.8 and 95.6 indicated four sugar units within 4 that were con®rmed as b-D-glucosyl units by the four anomeric proton signals at d 5.41 (1H, d, J 7:1 Hz), 5.04 (1H, d, J 7:6 Hz), 5.08 (1H, d, J 7:0 Hz) and 6.27 (1H, d, J 7:9 Hz). Therefore, 4 was elucidated as 3-O-[b-Dglucopyranosyl-(1 4 2)-b-D-glucopyranosyl]-hederagenin-28-O-[b-D-gluco-pyranosyl(1 4 6)-b-D-glucopyranosyl]ester by comparison with literature and it was ®rst isolated from Hedera taurica Carr. (Grishkovets et al., 1990).
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3. Experimental 3.1. General Melting points were measured with Yanaco MP-S3 melting point apparatus; optical rotations were taken on a P-E 241 MC polarimeter. IR spectra were taken on a Jasco FT/IR-230 IR spectrometer; NMR spectra were recorded on JEOL GX-400 and JEOL JNMLA400 Wb NMR spectrometers in C5D5N with TMS as standard; HR-FABMS and FABMS were measured with a JEOL JMS-700 mass analytical spectrometer; TLC and preparative TLC were carried out on precoated Kieselgel F254 plates (0.25 or 0.5 mm); silica gel and Cosmosil 75C18-OPN were employed for column chromatography and gel ®ltration was carried out on Sephadex LH-20. 3.2. Plant material The ¯ower buds of L. bournei Hemsl. (Caprifoliaceae) were purchased in Gansu Province of China and identi®ed by Associate Professor Yong-Hui Ding. A voucher specimen was deposited at Gansu Province Drug Control Institute. 3.3. Extraction and isolation The ¯ower buds of L. bournei Hemsl. (2 kg) were extracted with 90% ethanol (15 l 3). The ethanol solution was evaporated in vacuo to give the crude extract (LB-0, 1033.5 g) which was then suspended in water (1500 ml) and extracted with petroleum ether
Fig. 2. Signi®cant long-range correlation of bourneioside A (1) in HMBC-fg spectrum.
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(1500 ml 3), chloroform (2000 ml 3) and ethyl acetate (2000 ml 3), in the order given and successively, to give the corresponding petroleum ether (LB-1, 66.5 g), petroleum ether emulsion portion (LB-2, 8.5 g), chloroform (LB-3, 3.0 g) and ethyl acetate (LB-4, 102.2 g) extracts. The water soluble extract was next applied to a macroporous resin D 101 column and eluted with water, 30 and 90% ethanol, successively, to aord water (LB-5, 200.0 g), 30% ethanol (LB-6, 25.0 g) and 90% ethanol (LB-7, 30.0 g) eluates, respectively. The chloroform and ethyl acetate soluble portions were obtained and subjected to silica gel column chromatography, eluted with CHCl3/MeOH (0±100%) to aord 7 fractions: fr. 1, fr. 2 and fr. 3 were from 100:2 eluate; fr. 4, fr. 5, fr. 6 and fr. 7 were from 100:3, 100:6, 100:15 and 100:20 eluates, respectively. Fr. 7 was next applied to a Sephadex LH-20 column eluted with H2O/MeOH (0±100%). The 0±10% eluate from the Sephadex column was then applied to a Cosmosil 75C18-OPN column eluted with H2O/MeOH (0± 100%). The 60% eluate from the Cosmosil column was subjected to PTLC with CHCl3/MeOH/H2O (65:35:10, lower phase) to aord 1 Rf 0:40, 15.7 mg) and 3 Rf 0:38, 8.0 mg). The 90% eluate from the macroporous D 101 column was loaded onto a Sephadex LH-20 column which was eluted with H2O/ MeOH (0±100%). The 0±20% eluate from the Sephadex column was further subjected to the Cosmosil 75C18-OPN column eluted with H2O/acetone (0± 100%). The 30% eluate from the column was fractionated by PTLC with CHCl3/MeOH/H2O (7:7:1) to give 2 Rf 0:48, 12.6 mg) and 4 Rf 0:68, 12.0 mg), respectively. 3.4. Bourneioside A (3-O-b-D-glucopyranosyl-23hydroxy-lup-20(29)-en-28-oic acid-28-O-b-Dglucopyranosyl ester (1) White crystals (MeOH); mp 205±2078C; aD21 +55.08 (MeOH, c 0:1); HR-FABMS: m=z 819:4515 M Na]+, calcd. C42 H68 O14 Na for 819.4503; FABMS m/z: 819 M Na]+, 635 M H ÿ C6 H10 O5 ]+, 616 M ÿ C6 H12 O6 +, 518, 500; ÿ1 IR nKBr max cm : 3400, 2871, 1741, 1639, 1452, 1377, 1 1076, 890; H NMR (C5D5N, 400 MHz): aglycone moiety d 4.86 (1H, br.s, H-29), 4.71 (1H, br.s, H-29), 4.33 (1H, m, H-23), 4.28 (1H, m, H-3), 3.69 ( 1H, d, J 10:8 Hz, H-23), 3.40 (1H, dt, J 10:7, 4.4 Hz, H19), 2.67 (1H, m, H-13), 2.63 (1H, m, H-16), 2.29 (1H, m, H-2), 2.17 (1H, m, H-22), 2.09 (1H, m, H-21), 2.02 (1H, m, H-15), 1.92 (1H, m, H-2), 1.89 (1H, m, H-12), 1.73 (1H, m, H-18), 1.67 (1H, m, H-6), 1.58 (1H, m, H-1), 1.57 (1H, m, H-5), 1.51 (1H, m, H-7), 1.50 (1H, m, H-22), 1.46 (1H, m, H-16), 1.41 (1H, m, H-9), 1.38 (2H, m, H-11/H-21), 1.35 (1H, m, H-6), 1.29 (1H, m,
H-7), 1.19 (1H, m, H-12), 1.18 (1H, m, H-15), 1.17 (1H, m, H-11), 0.91 (1H, m, H-1), 1.72 (3H, s, H-30), 1.14 (3H, s, H-26), 0.99 (3H, s, H-27), 0.95 (3H, s, H24), 0.84 (3H, s, H-25); glucose attached to C-3 d 5.12 (1H, d, J 7:8 Hz, H-1 '), 4.50 (1H, dd, J 11:7, 2.2 Hz, H-6 '), 4.35 (2H, m, H-6 '/H-40), 4.22 (1H, m, H4 '), 4.14 (1H, m, H-3 '), 4.02 (1H, m, H-2 '), 3.87 (1H, m, H-5 '); glucose attached to C-28 d 6.42 (1H, d, J 8:0 Hz, H-10), 4.45 (1H, dd, J 12:4, 2.4 Hz, H-60), 4.39 (1H, m, H-60), 4.35 (2H, m, H-40/H-6 '), 4.28 (1H, m, H-30), 4.17 (1H, m, H-20), 4.03 (1H, m, H-50); 13C NMR (C5D5N, 100 MHz): aglycone moiety d 174.9 (C-28), 150.8 (C-20), 110.1 (C-29), 82.0 (C-3), 64.6 (C23), 57.0 (C-17), 50.9 (C-9), 49.8 (C-18), 47.7 (C-5), 47.5 (C-19), 43.5 (C-4), 42.8 (C-8), 41.2 (C-14), 39.0 (C-1), 38.3 (C-13), 37.0 (C-22), 36.9 (C-10), 34.3 (C-7), 32.2 (C-16), 30.9 (C-21), 30.1 (C-15), 26.0 (C-2/C-12), 21.1 (C-11), 18.1 (C-6), 19.4 (C-30), 16.9 (C-25), 16.4 (C-26), 14.9 (C-27), 13.5 (C-24); glucose attached to C3 d 105.8 (C-1 '), 78.7 (C-3 '), 78.3 (C-5 '), 75.9 (C-2 '), 71.6 (C-4 '), 62.8 (C-6 '); glucose attached to C-28 d 95.4 (C-10), 79.4 (C-50), 78.9 (C-30), 74.3 (C-20), 71.0 (C-40), 62.1 (C-60). 3.5. Bourneioside B (3-O-b-D-glucopyranosyl-23hydroxy-lup-20 (29)-en-28-oic acid-28-O-[b-Dglucopyranosyl-(1 4 6)-b-D-glucopyranosyl] ester (2) White crystals (MeOH), mp 214±2168C; aD21 +40.08 (MeOH, c 0:1); HR-FABMS: m=z 981:5036 M Na]+, calcd. C48 H78 O19 Na for 981.5035; FABMS m/z: 981 M Na]+, 766 M ÿ C6 H10 O5 ÿ CH2 O]+, 635 M H ÿ 2C6 H10 O5 ]+, ÿ1 613, 517, 501; IR nKBr max cm : 3400, 2870, 1736, 1640, 1 1450, 1377, 1078, 890; H NMR (C5D5N, 400 MHz): aglycone moiety d 4.84 (1H, br.s, H-29), 4.68 (1H, br.s, H-29), 4.31 (1H, m, H-23), 4.25 (1H, m, H-3), 3.68 ( 1H, d, J 10:8 Hz, H-23), 3.38 (1H, dt, J 10:9, 4.8 Hz, H-19), 2.63 (1H, m, H-13), 2.61 (1H, m, H-16), 2.27 (1H, m, H-2), 2.20 (1H, m, H-21/H-22), 1.98 (1H, m, H-15), 1.92 (1H, m, H-2), 1.85 (1H, m, H-12), 1.70 (1H, m, H-18), 1.65 (1H, m, H-6), 1.57 (2H, m, H-1/H5), 1.50 (1H, m, H-7), 1.46 (1H, m, H-16), 1.45 (1H, m, H-22), 1.40 (1H, m, H-9), 1.38 (3H, m, H-6/H-11/ H-21), 1.28 (1H, m, H-7), 1.20 (2H, m, H-11/H-12), 1.16 (1H, m, H-15), 0.93 (1H, m, H-1), 1.69 (3H, s, H30), 1.13 (3H, s, H-26), 0.97 (3H, s, H-27), 0.94 (3H, s, H-24), 0.85 (3H, s, H-25); glucose attached to C-3 d 5.10 (1H, d, J 8:0 Hz, H-1 '), 4.48 (1H, m, H-6 '), 4.36 (1H, m, H-6 '), 4.18 (1H, m, H-4 '), 4.12 (1H, m, H-3 '), 4.01 (1H, m, H-2 '), 3.86 (2H, m, H-5 '/H-51); glucose attached to C-28, inner glucose d 6.33 (1H, d, J 8:0 Hz, H-10), 4.69 (1H, d, J 10:7 Hz, H-60), 4.32 (1H, m, H-60), 4.28 (1H, m, H-40), 4.16 (2H, m, H-30/H-31), 4.10 (1H, m, H-50), 4.07 (1H, m, H-20); terminal glucose d 4.99 (1H, d, J 7:6 Hz, H-11), 4.46
T. Xiang et al. / Phytochemistry 54 (2000) 795±799
(1H, m, H-61), 4.34 (1H, m, H-61), 4.20 (1H, m, H41), 4.16 (2H, m, H-31/H-30), 3.97 (1H, m, H-21), 3.86 (1H, m, H-51/H-5'); 13C NMR (C5D5N, 100 MHz): aglycone moiety d 174.9 (C-28), 150.8 (C-20), 110.0 (C-29), 82.0 (C-3), 64.6 (C-23), 57.0 (C-17), 50.9 (C-9), 49.8 (C-18), 47.8 (C-5), 47.4 (C-19), 43.5 (C-4), 42.8 (C-8), 41.2 (C-14), 39.0 (C-1), 38.3 (C-13), 37.1 (C-22), 36.9 (C-10), 34.3 (C-7), 32.3 (C-16), 30.8 (C-21), 30.2 (C-15), 26.0 (C-2/C-12), 21.1 (C-11), 18.2 (C-6), 19.4 (C-30), 17.0 (C-25), 16.4 (C-26), 14.9 (C-27), 13.5 (C24); glucose attached to C-3 d 105.7 (C-1 '), 78.4 (C-3 '), 78.3 (C-5 '), 75.9 (C-2 '), 71.6 (C-4 '), 62.8 (C-6 '); glucose attached to C-28, inner glucose d 95.2 (C-10), 78.7 (C-30/C-31), 78.0 (C-50), 74.0 (C-20), 70.9 (C-40), 69.5 (C-60); terminal glucose d 105.4 (C-11), 78.7 (C-31/C30), 78.3 (C-51), 75.1 (C-21), 71.5 (C-41), 62.6 (C-61). 3.6. 3-O-b-D-glucopyranosyl-hederagenin-28-O-b-Dglucopyranosyl ester (3) White crystals (MeOH); mp 211±2138C; and the NMR data were assigned by 1H NMR, 13C NMR and comparison with literature values (Takemoto et al., 1984). 3.7. 3-O-[b-D-glucopyranosyl-(1 4 2)-b-Dglucopyranosyl]-hederagenin-28-O-[b-D-glucopyranosyl (1 4 6)-b-D-glucopyranosyl]ester (4) White crystals (MeOH); mp 146±1488C; and the
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NMR data were assigned by 1H NMR, 13C NMR and comparison with literature values (Grishkovets et al., 1990).
References Chang, Ch. W., Lin, M.T., Lee, Sh.Sh., Liu, K.C., Hsu, F.L., Lin, J.Y., 1995. Dierential inhibition of reverse transcriptase and cellular DNA polymerase-a activities by lignans isolated from Chinese Herbs, Phyllanthus myrtifolius Moon and tannis from Lonicera japonica Thunb. and Castanopsis hystrix. Antiviral Res. 27 (4), 367±374. Grishkovets, V.I., Loloiko, A.A., Shashkov, A.S., Chirva, V.Ya., 1990. Triterpene glucosides of Hedera taurica VI. Structures of hederosides G, H1, H2 and I from the berries of Crimen Ivy. Khim Prir Soedin 6, 779±783. Ikuta, A., Itokawa, H., 1988. Triterpenes of Paeonia japonica Callus tissue. Phytochemistry 27 (9), 2813±2815. Lou, H.X., Tian, J.G., Yuan, H.Q., Ji, M., 1995. Triterpenoids saponins from ¯os Lonicera japonica. Chinese Journal of Medicinal Chemistry 5 (3), 208±209. Nakatani, M., Miyazaki, Y., Iwashita, T., Naoki, H., Hase, T., 1989. Triterpenes from Ilex rotunda fruits. Phytochemistry 28 (5), 1479±1482. Takemoto, T., Arihara, S., Yoshikawa, K., Kusumoto, K., Yano, I., Hayashi, T., 1984. Studies on the constituents of cucurbitaceae plants. VI. On the saponin constituents of Lua cylindrica Roem. Yakugaku Zasshi 104 (3), 246±255. Ye, W.C., Ji, N.N., Zhao, S.X., Liu, J.H., Ye, T., Mckervey, M.A., Stevenson, P., 1996. Triterpenoids from Pulsatilla chinensis. Phytochemisty 42 (3), 799±802.
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Saponins from Lonicera bournei Ting Xiang a, Yasuhiro Tezuka b, Li-Jun Wu a,*, Arjun H. Banskota b, Shigetoshi Kadota b a
Department of Natural Medicines, Shenyang Pharmaceutical University, Shenyang 110015, People's Republic of China b Institute of Natural Medicine, Toyama Medical and Pharmaceutical University, Toyama, Japan Received 13 October 1999; received in revised form 28 February 2000
Abstract The lupane-triterpene glycosides, bourneioside A and bourneioside B, and two known saponins were isolated from Lonicera bournei Hemsl. The structures of bourneioside A and B were elucidated as 3-O-b-D-glucopyranosyl-23-hydroxy-lup-20(29)-en-28oic acid-28-O-b-D-glucopyranosyl ester and 3-O-b-D-glucopyranosyl-23-hydroxy-lup-20(29)-en-28-oic acid-28-O-[b-Dglucopyranosyl-(1 4 6)-b-D-glucopyranosyl] ester, respectively, on the basis of spectral data and chemical evidence. 7 2000 Published by Elsevier Science Ltd. Keywords: Lonicera bournei Hemsl; Caprifoliaceae; Bourneioside A; Bourneioside B
1. Introduction
2. Results and discussion
The ¯ower buds of Lonicera bournei Hemsl. and L. japonica Thunb. are widely used in northwest and southwest China. Both are used as antibacterial, antiin¯ammatory agents, as well as folk medicines to treat encephalitis, in¯uenza, fever, pneumonia, dysentery, enteritis and so on. Several tannins and triterpenes from L. japonica Thunb. showed inhibitory eect on HIV-1 RT and a cytoprotective eect against CCl4induced hepatic injury, respectively (Chang et al., 1995; Lou et al., 1995). This paper reports the isolation and structure elucidation of two new lupane-triterpene glycosides named bourneioside A (1) and bourneioside B (2) along with two known saponins (3 and 4) from L. bournei Hemsl. No correlation with these compounds to any medicinal property was made in this study.
The ¯ower buds of L. bournei Hemsl. were extracted with 90% ethanol. The extract was further isolated as described in Section 3 to provide two new lupane-triterpene glycosides 1 (15.7 mg) and 2 (12.6 mg) (see Fig. 1) together with two known triterpenoid glycosides, 3-O-b-D-glucopyranosyl-hederagenin-28-O-b-Dglucopyranosyl ester 3 (8.0 mg) and 3-O-[b-D-glucopyranosyl-(1 4 2)-b-D-glucopyranosyl]-hederagenin-28-O[b-D-glucopyranosyl(1 4 6)-b-D-glucopyranosyl]ester 4 (12.0 mg). Bourneioside A (1) was obtained as white crystals (MeOH) with mp 205±2078C, having aD21 +55.08 (MeOH, c 0:1), and it responded positively to the Liebermann-Burchard and Molisch tests. HR-FABMS gave a M Na]+ peak at m/z 819.4515 corresponding to the molecular formula of C42H68O14. The IR spectrum showed the presence of hydroxyl (3400 cmÿ1), carboxyl (1741 cmÿ1) and exo-methylene (1639, 890 cmÿ1) groups. The 13C NMR and DEPT spectra of the aglycone moiety exhibited 30 carbon signals (seven quaternary, six methine, 12 methylene and ®ve methyl carbons). The presence of a methyl group attached to a terminal double bond was con®rmed by the 1H
* Corresponding author. Department of Natural Medicines, Faculty of Traditional Chinese Medicine, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang, 110015, People's Republic of China. E-mail address: [email protected] (T. Xiang).
0031-9422/00/$ - see front matter 7 2000 Published by Elsevier Science Ltd. PII: S 0 0 3 1 - 9 4 2 2 ( 0 0 ) 0 0 1 9 4 - 1
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NMR signals at d 4.86 (1H, br.s ), 4.71 (1H, br.s ), 1.72 (3H, s ) and the 13C NMR signals at d 150.8, 110.1, 19.4. One primary hydroxyl group was assigned to the C-23 position based on the presence of hydroxy methylene signals at d 4.33 (1H, m ), 3.69 (1H, d, J 10:8 Hz) and d 64.6. The carbonyl carbon signal observed at d 174.9 was assigned to C-28. The above data suggested that 1 was a derivative of 23-hydroxylupanetriterpene (Nakatani et al., 1989; Ikuta and Itokawa, 1988; Ye et al., 1996). Acid hydrolysis conducted on TLC yielded only glucose. The anomeric carbon signals at d 105.8 and 95.4 indicated two glucosyl units within 1 that were con®rmed as b-D-glucosyl units by the two anomeric proton signals at d 5.12 (1H, d, J 7:8 Hz) and 6.42 (1H, d, J 8:0 Hz). The TOCSY spectrum demonstrated that the proton signals at d 5.12, 4.50 (dd, J 11:7, 2.2 Hz), 4.35 (m ), 4.22 (m ), 4.14 (m ), 4.02 (m ), 3.87 (m ) and d 6.42, 4.45 (dd, J 12:4, 2.4 Hz), 4.39 (m ), 4.35 (m ), 4.28 (m ), 4.17 (m ), 4.03 (m ) were due to two dierent glucoses, respectively. In the 1H±1H COSY spectrum, correlation peaks could be observed from the following pairs: d 5.12 (H-1')/4.02 (H-2 '), 4.02 (H2 ')/4.14 (H-3'), 4.14 (H-3 ')/4.22 (H-4'), 4.22 (H-4 ')/ 3.87 (H-5 '), 3.87 (H-5 ')/4.50, 4.35 (H-6 '); d 6.42 (H10)/4.17 (H-20), 4.17 (H-20)/4.28 (H-30), 4.28 (H-30)/ 4.35 (H-40), 4.35 (H-40)/4.03 (H-50), 4.03 (H-50)/4.45, 4.39 (H-60). Thus, the proton signals of the sugar moieties were assigned one by one. Furthermore, the carbon signals of the sugar moieties were identi®ed by means of HMQC-fg spectrum. As shown in Fig. 2, the long-range correlations between d 5.12 (H-1 ') and 82.0 (C-3), as well as d 6.42
Fig. 1. Structures of bourneioside A (1) and bourneioside B (2) from L. bournei Hemsl.
(H-10) and 174.9 (C-28) suggested that the two glucosyl units were attached to C-3 and C-28 of the aglycone, respectively. Therefore, compound 1 was established as 3-O-b-D-glucopyranosyl-23-hydroxy-lup20(29)-en-28-oic acid-28-O-b-D-glucopyranosyl ester, named bourneioside A. Bourneioside B (2) was obtained as white crystals (MeOH) with mp 214±2168C, having aD21 +40.08 (MeOH, c 0:1), and it also responded positively to the Liebermann-Burchard and Molisch tests. HRFABMS showed M Na]+ peak at m/z 981.5036 corresponding to molecular formula of C48H78O19. The 1 H NMR and 13C NMR spectra of the aglycone moiety of 2 were similar to those of 1, which indicated that 2 was also a 23-hydroxylupane-triterpene derivative. Acid hydrolysis carried out on TLC yielded only glucose. The anomeric carbon signals at d 105.7, 105.4 and 95.2 indicated three glucosyl units within 2 that were con®rmed as b-D-glucosyl units by the three corresponding anomeric proton signals at d 5.10 (1H, d, J 8:0 Hz), 6.33 (1H, d, J 8:0 Hz), 4.99 (1H, d, J 7:6 Hz). The TOCSY spectrum revealed that the proton signals at d 5.10, 4.48 (m ), 4.36 (m ), 4.18 (m ), 4.12 (m ), 4.01 (m ), 3.86 (m ); d 6.33, 4.69 (d, J 10:7 Hz), 4.32 (m ), 4.28 (m ), 4.16 (m ), 4.10 (m ), 4.07 (m ) and d 4.99, 4.46 (m ), 4.34 (m ), 4.20 (m ), 4.16 (m ), 3.97 (m ), 3.86 (m ) were due to three dierent glucoses, respectively. In the 1Hÿ1H COSY spectrum, the correlation peaks could be observed from the following pairs: d 5.10 (H-1 ')/4.01 (H-2 '), 4.01 (H-2 ')/4.12 (H3 '), 4.12 (H-3 ')/4.18 (H-4'), 4.18 (H-4 ')/3.86 (H-5 '), 3.86 (H-5 ')/4.48, 4.36 (H-6'); d 6.33 (H-10)/4.07 (H-20), 4.07 (H-20)/4.16 (H-30), 4.16 (H-30)/4.28 (H-40), 4.28 (H-40)/4.10 (H-50), 4.10 (H-50)/4.69, 4.32 (H-60); d 4.99 (H-11)/3.97 (H-21), 3.97 (H-21)/4.16 (H-31), 4.16 (H31)/4.20 (H-41), 4.20 (H-41)/3.86 (H-51), 3.86 (H-51)/ 4.46, 4.34 (H-61). These analyses clari®ed the assignments of the proton signals of sugar moieties. The carbon signals of the sugar moieties were identi®ed by means of HMQC-fg spectrum. The anomeric proton signals at d 6.33 (H-10), 5.10 (H-1'), 4.99 (H-11) showed cross-peaks with the carbon signals at d 174.9 (C-28), 82.0 (C-3), 69.5 (C-60) in the HMBC-fg spectrum, respectively. The carbon signal at d 69.5 showed correlation with proton signals at d 4.69 (1H, d, J 10:7 Hz) and 4.32 (1H, m ) which were determined to be H-60 of the glucosyl moiety attached to C-28 by TOCSY spectrum. Thus, compound 2 was established as a new 23-hydroxylupanetriterpene saponin, 3-O-b-D-glucopyranosyl-23hydroxy-lup-20(29)-en-28-oic acid-28-O-[b-D-glucopyranosyl-(1 4 6)-b-D-glucopyranosyl] ester, named bourneioside B. Compound 3 was obtained as white crystals (MeOH) with mp 211±2138C, and it responded
T. Xiang et al. / Phytochemistry 54 (2000) 795±799
positively to the Liebermann-Burchard and Molisch tests. Acid hydrolysis conducted on TLC yielded only glucose. The 1H NMR and the 13C NMR spectral data revealed that the aglycone moiety of 3 was hedergenin (Takemoto et al., 1984). The anomeric signals at d 105.8 and 95.7 indicated two glucosyl units within 3 that were con®rmed as b-D-glucosyl units by the two anomeric proton signals at d 5.11 (1H, d, J 7:8 Hz), 6.32 (1H, d, J 7:8 Hz). Thus, 3 was elucidated as 3-O-b-D-glucopyranosyl-hederagenin-28-O-bD-glucopyranosyl ester by comparison with literature and it was ®rst isolated from Lua cylindrica Roem. (Takemoto et al., 1984). Compound 4 was obtained as white crystals (MeOH) with mp 146±1488C, and it responded positively to the Liebermann-Burchard and Molisch tests. Acid hydrolysis conducted on TLC yielded only glucose. The 1H NMR and the 13C NMR spectral data of the aglycone moiety of 4 were similar to those of 3, which indicated that the aglycone moiety of 4 was also hedergenin. The sugar moieties of 4 were dierent from those of 3. The anomeric signals at d 105.9, 105.3, 103.8 and 95.6 indicated four sugar units within 4 that were con®rmed as b-D-glucosyl units by the four anomeric proton signals at d 5.41 (1H, d, J 7:1 Hz), 5.04 (1H, d, J 7:6 Hz), 5.08 (1H, d, J 7:0 Hz) and 6.27 (1H, d, J 7:9 Hz). Therefore, 4 was elucidated as 3-O-[b-Dglucopyranosyl-(1 4 2)-b-D-glucopyranosyl]-hederagenin-28-O-[b-D-gluco-pyranosyl(1 4 6)-b-D-glucopyranosyl]ester by comparison with literature and it was ®rst isolated from Hedera taurica Carr. (Grishkovets et al., 1990).
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3. Experimental 3.1. General Melting points were measured with Yanaco MP-S3 melting point apparatus; optical rotations were taken on a P-E 241 MC polarimeter. IR spectra were taken on a Jasco FT/IR-230 IR spectrometer; NMR spectra were recorded on JEOL GX-400 and JEOL JNMLA400 Wb NMR spectrometers in C5D5N with TMS as standard; HR-FABMS and FABMS were measured with a JEOL JMS-700 mass analytical spectrometer; TLC and preparative TLC were carried out on precoated Kieselgel F254 plates (0.25 or 0.5 mm); silica gel and Cosmosil 75C18-OPN were employed for column chromatography and gel ®ltration was carried out on Sephadex LH-20. 3.2. Plant material The ¯ower buds of L. bournei Hemsl. (Caprifoliaceae) were purchased in Gansu Province of China and identi®ed by Associate Professor Yong-Hui Ding. A voucher specimen was deposited at Gansu Province Drug Control Institute. 3.3. Extraction and isolation The ¯ower buds of L. bournei Hemsl. (2 kg) were extracted with 90% ethanol (15 l 3). The ethanol solution was evaporated in vacuo to give the crude extract (LB-0, 1033.5 g) which was then suspended in water (1500 ml) and extracted with petroleum ether
Fig. 2. Signi®cant long-range correlation of bourneioside A (1) in HMBC-fg spectrum.
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(1500 ml 3), chloroform (2000 ml 3) and ethyl acetate (2000 ml 3), in the order given and successively, to give the corresponding petroleum ether (LB-1, 66.5 g), petroleum ether emulsion portion (LB-2, 8.5 g), chloroform (LB-3, 3.0 g) and ethyl acetate (LB-4, 102.2 g) extracts. The water soluble extract was next applied to a macroporous resin D 101 column and eluted with water, 30 and 90% ethanol, successively, to aord water (LB-5, 200.0 g), 30% ethanol (LB-6, 25.0 g) and 90% ethanol (LB-7, 30.0 g) eluates, respectively. The chloroform and ethyl acetate soluble portions were obtained and subjected to silica gel column chromatography, eluted with CHCl3/MeOH (0±100%) to aord 7 fractions: fr. 1, fr. 2 and fr. 3 were from 100:2 eluate; fr. 4, fr. 5, fr. 6 and fr. 7 were from 100:3, 100:6, 100:15 and 100:20 eluates, respectively. Fr. 7 was next applied to a Sephadex LH-20 column eluted with H2O/MeOH (0±100%). The 0±10% eluate from the Sephadex column was then applied to a Cosmosil 75C18-OPN column eluted with H2O/MeOH (0± 100%). The 60% eluate from the Cosmosil column was subjected to PTLC with CHCl3/MeOH/H2O (65:35:10, lower phase) to aord 1 Rf 0:40, 15.7 mg) and 3 Rf 0:38, 8.0 mg). The 90% eluate from the macroporous D 101 column was loaded onto a Sephadex LH-20 column which was eluted with H2O/ MeOH (0±100%). The 0±20% eluate from the Sephadex column was further subjected to the Cosmosil 75C18-OPN column eluted with H2O/acetone (0± 100%). The 30% eluate from the column was fractionated by PTLC with CHCl3/MeOH/H2O (7:7:1) to give 2 Rf 0:48, 12.6 mg) and 4 Rf 0:68, 12.0 mg), respectively. 3.4. Bourneioside A (3-O-b-D-glucopyranosyl-23hydroxy-lup-20(29)-en-28-oic acid-28-O-b-Dglucopyranosyl ester (1) White crystals (MeOH); mp 205±2078C; aD21 +55.08 (MeOH, c 0:1); HR-FABMS: m=z 819:4515 M Na]+, calcd. C42 H68 O14 Na for 819.4503; FABMS m/z: 819 M Na]+, 635 M H ÿ C6 H10 O5 ]+, 616 M ÿ C6 H12 O6 +, 518, 500; ÿ1 IR nKBr max cm : 3400, 2871, 1741, 1639, 1452, 1377, 1 1076, 890; H NMR (C5D5N, 400 MHz): aglycone moiety d 4.86 (1H, br.s, H-29), 4.71 (1H, br.s, H-29), 4.33 (1H, m, H-23), 4.28 (1H, m, H-3), 3.69 ( 1H, d, J 10:8 Hz, H-23), 3.40 (1H, dt, J 10:7, 4.4 Hz, H19), 2.67 (1H, m, H-13), 2.63 (1H, m, H-16), 2.29 (1H, m, H-2), 2.17 (1H, m, H-22), 2.09 (1H, m, H-21), 2.02 (1H, m, H-15), 1.92 (1H, m, H-2), 1.89 (1H, m, H-12), 1.73 (1H, m, H-18), 1.67 (1H, m, H-6), 1.58 (1H, m, H-1), 1.57 (1H, m, H-5), 1.51 (1H, m, H-7), 1.50 (1H, m, H-22), 1.46 (1H, m, H-16), 1.41 (1H, m, H-9), 1.38 (2H, m, H-11/H-21), 1.35 (1H, m, H-6), 1.29 (1H, m,
H-7), 1.19 (1H, m, H-12), 1.18 (1H, m, H-15), 1.17 (1H, m, H-11), 0.91 (1H, m, H-1), 1.72 (3H, s, H-30), 1.14 (3H, s, H-26), 0.99 (3H, s, H-27), 0.95 (3H, s, H24), 0.84 (3H, s, H-25); glucose attached to C-3 d 5.12 (1H, d, J 7:8 Hz, H-1 '), 4.50 (1H, dd, J 11:7, 2.2 Hz, H-6 '), 4.35 (2H, m, H-6 '/H-40), 4.22 (1H, m, H4 '), 4.14 (1H, m, H-3 '), 4.02 (1H, m, H-2 '), 3.87 (1H, m, H-5 '); glucose attached to C-28 d 6.42 (1H, d, J 8:0 Hz, H-10), 4.45 (1H, dd, J 12:4, 2.4 Hz, H-60), 4.39 (1H, m, H-60), 4.35 (2H, m, H-40/H-6 '), 4.28 (1H, m, H-30), 4.17 (1H, m, H-20), 4.03 (1H, m, H-50); 13C NMR (C5D5N, 100 MHz): aglycone moiety d 174.9 (C-28), 150.8 (C-20), 110.1 (C-29), 82.0 (C-3), 64.6 (C23), 57.0 (C-17), 50.9 (C-9), 49.8 (C-18), 47.7 (C-5), 47.5 (C-19), 43.5 (C-4), 42.8 (C-8), 41.2 (C-14), 39.0 (C-1), 38.3 (C-13), 37.0 (C-22), 36.9 (C-10), 34.3 (C-7), 32.2 (C-16), 30.9 (C-21), 30.1 (C-15), 26.0 (C-2/C-12), 21.1 (C-11), 18.1 (C-6), 19.4 (C-30), 16.9 (C-25), 16.4 (C-26), 14.9 (C-27), 13.5 (C-24); glucose attached to C3 d 105.8 (C-1 '), 78.7 (C-3 '), 78.3 (C-5 '), 75.9 (C-2 '), 71.6 (C-4 '), 62.8 (C-6 '); glucose attached to C-28 d 95.4 (C-10), 79.4 (C-50), 78.9 (C-30), 74.3 (C-20), 71.0 (C-40), 62.1 (C-60). 3.5. Bourneioside B (3-O-b-D-glucopyranosyl-23hydroxy-lup-20 (29)-en-28-oic acid-28-O-[b-Dglucopyranosyl-(1 4 6)-b-D-glucopyranosyl] ester (2) White crystals (MeOH), mp 214±2168C; aD21 +40.08 (MeOH, c 0:1); HR-FABMS: m=z 981:5036 M Na]+, calcd. C48 H78 O19 Na for 981.5035; FABMS m/z: 981 M Na]+, 766 M ÿ C6 H10 O5 ÿ CH2 O]+, 635 M H ÿ 2C6 H10 O5 ]+, ÿ1 613, 517, 501; IR nKBr max cm : 3400, 2870, 1736, 1640, 1 1450, 1377, 1078, 890; H NMR (C5D5N, 400 MHz): aglycone moiety d 4.84 (1H, br.s, H-29), 4.68 (1H, br.s, H-29), 4.31 (1H, m, H-23), 4.25 (1H, m, H-3), 3.68 ( 1H, d, J 10:8 Hz, H-23), 3.38 (1H, dt, J 10:9, 4.8 Hz, H-19), 2.63 (1H, m, H-13), 2.61 (1H, m, H-16), 2.27 (1H, m, H-2), 2.20 (1H, m, H-21/H-22), 1.98 (1H, m, H-15), 1.92 (1H, m, H-2), 1.85 (1H, m, H-12), 1.70 (1H, m, H-18), 1.65 (1H, m, H-6), 1.57 (2H, m, H-1/H5), 1.50 (1H, m, H-7), 1.46 (1H, m, H-16), 1.45 (1H, m, H-22), 1.40 (1H, m, H-9), 1.38 (3H, m, H-6/H-11/ H-21), 1.28 (1H, m, H-7), 1.20 (2H, m, H-11/H-12), 1.16 (1H, m, H-15), 0.93 (1H, m, H-1), 1.69 (3H, s, H30), 1.13 (3H, s, H-26), 0.97 (3H, s, H-27), 0.94 (3H, s, H-24), 0.85 (3H, s, H-25); glucose attached to C-3 d 5.10 (1H, d, J 8:0 Hz, H-1 '), 4.48 (1H, m, H-6 '), 4.36 (1H, m, H-6 '), 4.18 (1H, m, H-4 '), 4.12 (1H, m, H-3 '), 4.01 (1H, m, H-2 '), 3.86 (2H, m, H-5 '/H-51); glucose attached to C-28, inner glucose d 6.33 (1H, d, J 8:0 Hz, H-10), 4.69 (1H, d, J 10:7 Hz, H-60), 4.32 (1H, m, H-60), 4.28 (1H, m, H-40), 4.16 (2H, m, H-30/H-31), 4.10 (1H, m, H-50), 4.07 (1H, m, H-20); terminal glucose d 4.99 (1H, d, J 7:6 Hz, H-11), 4.46
T. Xiang et al. / Phytochemistry 54 (2000) 795±799
(1H, m, H-61), 4.34 (1H, m, H-61), 4.20 (1H, m, H41), 4.16 (2H, m, H-31/H-30), 3.97 (1H, m, H-21), 3.86 (1H, m, H-51/H-5'); 13C NMR (C5D5N, 100 MHz): aglycone moiety d 174.9 (C-28), 150.8 (C-20), 110.0 (C-29), 82.0 (C-3), 64.6 (C-23), 57.0 (C-17), 50.9 (C-9), 49.8 (C-18), 47.8 (C-5), 47.4 (C-19), 43.5 (C-4), 42.8 (C-8), 41.2 (C-14), 39.0 (C-1), 38.3 (C-13), 37.1 (C-22), 36.9 (C-10), 34.3 (C-7), 32.3 (C-16), 30.8 (C-21), 30.2 (C-15), 26.0 (C-2/C-12), 21.1 (C-11), 18.2 (C-6), 19.4 (C-30), 17.0 (C-25), 16.4 (C-26), 14.9 (C-27), 13.5 (C24); glucose attached to C-3 d 105.7 (C-1 '), 78.4 (C-3 '), 78.3 (C-5 '), 75.9 (C-2 '), 71.6 (C-4 '), 62.8 (C-6 '); glucose attached to C-28, inner glucose d 95.2 (C-10), 78.7 (C-30/C-31), 78.0 (C-50), 74.0 (C-20), 70.9 (C-40), 69.5 (C-60); terminal glucose d 105.4 (C-11), 78.7 (C-31/C30), 78.3 (C-51), 75.1 (C-21), 71.5 (C-41), 62.6 (C-61). 3.6. 3-O-b-D-glucopyranosyl-hederagenin-28-O-b-Dglucopyranosyl ester (3) White crystals (MeOH); mp 211±2138C; and the NMR data were assigned by 1H NMR, 13C NMR and comparison with literature values (Takemoto et al., 1984). 3.7. 3-O-[b-D-glucopyranosyl-(1 4 2)-b-Dglucopyranosyl]-hederagenin-28-O-[b-D-glucopyranosyl (1 4 6)-b-D-glucopyranosyl]ester (4) White crystals (MeOH); mp 146±1488C; and the
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NMR data were assigned by 1H NMR, 13C NMR and comparison with literature values (Grishkovets et al., 1990).
References Chang, Ch. W., Lin, M.T., Lee, Sh.Sh., Liu, K.C., Hsu, F.L., Lin, J.Y., 1995. Dierential inhibition of reverse transcriptase and cellular DNA polymerase-a activities by lignans isolated from Chinese Herbs, Phyllanthus myrtifolius Moon and tannis from Lonicera japonica Thunb. and Castanopsis hystrix. Antiviral Res. 27 (4), 367±374. Grishkovets, V.I., Loloiko, A.A., Shashkov, A.S., Chirva, V.Ya., 1990. Triterpene glucosides of Hedera taurica VI. Structures of hederosides G, H1, H2 and I from the berries of Crimen Ivy. Khim Prir Soedin 6, 779±783. Ikuta, A., Itokawa, H., 1988. Triterpenes of Paeonia japonica Callus tissue. Phytochemistry 27 (9), 2813±2815. Lou, H.X., Tian, J.G., Yuan, H.Q., Ji, M., 1995. Triterpenoids saponins from ¯os Lonicera japonica. Chinese Journal of Medicinal Chemistry 5 (3), 208±209. Nakatani, M., Miyazaki, Y., Iwashita, T., Naoki, H., Hase, T., 1989. Triterpenes from Ilex rotunda fruits. Phytochemistry 28 (5), 1479±1482. Takemoto, T., Arihara, S., Yoshikawa, K., Kusumoto, K., Yano, I., Hayashi, T., 1984. Studies on the constituents of cucurbitaceae plants. VI. On the saponin constituents of Lua cylindrica Roem. Yakugaku Zasshi 104 (3), 246±255. Ye, W.C., Ji, N.N., Zhao, S.X., Liu, J.H., Ye, T., Mckervey, M.A., Stevenson, P., 1996. Triterpenoids from Pulsatilla chinensis. Phytochemisty 42 (3), 799±802.