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A new aristolactam alkaloid from the stems of Dasymaschalon trichophorum
Chinese Journal of Natural Medicines 2013, 11(1): 0081−0083
Chinese Journal of Natural Medicines
A new aristolactam alkaloid from the stems of Dasymaschalon trichophorum ZHOU Xiao-Lei1, 2, WU Jiu-Hong1*, BAI Jiao2, HU Xiao-Lan2, LI En-Zhen1, 3, SHI Ning1, PEI Yue-Hu2 1
Department of Pharmacy, The 306th hospital of PLA, Beijing 100101, China; School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang 110016, China; 3 Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Beijing 100850, China 2
Available online 20 Jan. 2013 [ABSTRACT] AIM: To study the chemical constituents of the stem of Dasymaschaon trichophorum Merr.. METHODS: The compounds were isolated and purified by the methods of various chromatography. Their structures were elucidated on the basis of spectroscopic methods. RESULTS: Five aristolactam alkaloids were obtained and their structures were identified as 10-amino-3,6-dihydroxy-2,4-dimethoxyphenanthrene-1-carboxylic acid lactam (1), enterocarpam-II (2), oldhamactam (3), goniopedaline (4), and stigmalactam (5), respectively. CONCLUSION: Compound 1 is a new aristolactam alkaloid, while compounds 2−5 are reported from this genus for the first time. [KEY WORDS] Annonaceae; Dasymaschaon trichophorum; Aristolactam alkaloids
[CLC Number] R284.1
1
[Document code] A
[Article ID] 1672-3651(2013)01-0081-03
Introduction
The genus Dasymaschalon Dalla Torre & Harms comprises about 16 species, three of which are distributed in the south of China[1]. To date, only a few chemical constituents of this genus have been reported, and most of them were flavonoids and alkaloids[2-5]. In a phytochemical study on the specie Dasymaschalon trichophorum Merr., five aristolactam alkaloids were isolated and their structures were elucidated on the basis of spectroscopic analysis as 10-amino-3, 6-dihydroxy-2, 4-dimethoxyphenanthrene-1-carboxylic acid lactam (1), a new alkaloid, and four known aristolactam alkaloids, enterocarpam-II (2), oldhamactam (3), goniopedaline (4), and stigmalactam (5).
2
Results and Discussion
Alkaloid 1 was isolated as brownish yellow needles. The molecular formula C17H13O5N was determined by
[Received on] 11-Apr.-2012 [Research funding] This project was supported by the National Natural Science Foundation of China (No.81072546) and the Chinese PLA National Science Fund for Distinguished Young Scholars Grant during the 11th “Five-Year” Plan period (No.06J001). [*Corresponding author] WU Jiu-Hong: Prof., Tel: 86-1066354564; Fax: 86-10-64879825, Email: [email protected] These authors have no conflict of interest to declare. Copyright © 2013, China Pharmaceutical University. Published by Elsevier B.V. All rights reserved
HR-ESI-MS at m/z 312.0867 [M + H]+ (Calcd. for C17H14O5 N1 312.086 6 ). The 13C NMR spectrum of alkaloid 1 indicated the presence of 15 sp2 carbons (including a carbonyl carbon at δ 166.9), which are typical for an aristolactam alkaloid[5-7]. Therefore, it was determined that 1 possesses an aristolactam skeleton. Comparing its 1H and 13C NMR data (see Table 1) with those of the known alkaloid goniopedaline (4), 1 had an additional hydroxyl group. In the 1H-NMR spectrum of 1, an ABX system for aromatic protons was observed at δ 8.52 (1H, d, J = 2.5 Hz), 7.75 (1H, d, J = 8.6 Hz), and 7.04 (1H, dd, J = 8.6, 2.5 Hz), attributable to H-5, H-8, and H-7, respectively. A singlet at δ 7.05 (1H) was assigned to H-9. These data resembled those of 5, 7, 8, 9-unsubstituted aristolactams. The HMBC correlations (see Table 1) between H-5 to C-6 and H-8 to C-6 confirmed the assignment of one hydroxyl OH (δ 9.65) at C-6. The remaining two methoxyl signals at δ 4.01 (3H, s) and 4.33 (3H, s) were assigned to C-4 and C-2. The phenolic hydroxyl (δ 9.36) could be assigned to C-3. These assignments were verified by HMBC correlations of the methoxyl protons at δ 4.01 with δ 150.1 (C-2), and δ 4.33 with δ 148.8 (C-4). The singlet signal at δ 10.73 was attributed to the amide NH, and this was confirmed by HMBC correlations between δ 10.73 to δ 109.9 (C-1), δ 132.4 (C-10) and δ 122.2 (C-10a). On the basis of these results, 1 was established structurally as 10-amino-3, 6dihydroxy-2, 4-dimethoxyphenanthrene-1-carb-oxylic acid lactam The structures of the known alkaloids (2−5) were identified by comparing their spectroscopic data (1H NMR and 13C
ZHOU Xiao-Lei, et al. /Chinese Journal of Natural Medicines 2013, 11(1): 81−83
NMR) with those reported in the literature.
4
3
10-Amino-3, 6-dihydroxy-2, 4- dimethoxyphenanthrene-1-carboxylic acid lactam (1) Brownish yellow needles (MeOH), HR-ESI-MS m/z 312.086 7 [M + H]+ (Calcd. for C17H14O5 N1, 312.086 6 ); 1H and 13C NMR data are shown in Table 1.
Experimental
3.1 General experimental procedures HR-ESI-MS was recorded on a Varian QFT-ESI mass spectrometer. 1H NMR and 13C NMR spectra were recorded on Bruker AV-600 NMR spectrometers using TMS as an internal standard. The chromatographic silica gel (54−74 μm) was purchased from Qingdao Marine Chemical Factory (Qingdao, China). Sephadex LH-20 was purchased from GE Healthcare. RP-HPLC analysis and semi-preparation were conducted using Hitachi L-6000 pumping system equipped with a Hitachi L-7400 UV detector and performed with a C18 column (10 mm × 250 mm, GL science Co., Ltd., Japan). Fractions were monitored by thin layer chromatography (TLC), and spots were detected with a UV 254 nm lamp and by spraying with 10% H2SO4 in EtOH followed by heating at 105 ºC for 5 min. 3.2 Plant material The stems of Dasymaschaon trichophorum Merr. were collected in Dec. 2010 in Beihai, Guangxi Zhuang Autonomous Region, China, and identified by Prof. FANG Ding (Guangxi Institute of Chinese Medicine & Pharmaceutical Sciences). A voucher specimen (No. 76773) was deposited in The 306th Hospital of PLA. 3.3 Extraction and isolation The dried and powered plant material (8.1 kg) was extracted with 95% EtOH three times under reflux. After concentration, the extract (750 g) was suspended in water and partitioned with CHCl3 followed by EtOAc. The CHCl3 residue (15 g) was subjected to silica gel column chromatography (600 g) and eluted with petroleum−EtOAc mixtures in a gradient (1 : 0 →0 : 1) to yield 15 fractions (1−15) . Fraction 10 was then subjected to column chromatography over silica gel (90 g) eluted with CHCl3−MeOH (100 : 1 →100 : 5) to give subfractions (Fr. A1−Fr. A5). Fr. A2 was purified by Sephadex LH-20 (MeOH), followed by RP-HPLC [MeOH/H2O (58 : 42), 4.0 mL·min−1] to obtain 4 (2.3 mg). Fr. A3 was further separated by silica gel chromatography, followed by Sephadex LH-20 column chromatography to obtain 3 (12.5 mg). Fr. A4 was purified by Sephadex LH-20 (MeOH), followed by RP-HPLC [MeOH/H2O (54 : 46), 4.0 mL·min−1] to obtain 5 (3.7 mg). Fraction 11 was subjected to column chromatography over silica gel (55 g) eluted with CHCl3−MeOH (9 : 2) to give subfractions (Fr. B1-Fr. B4). Fr. B4 was purified by Sephadex LH-20 (MeOH), followed by RP-HPLC [MeOH/H2O (52: 48), 4.0 mL·min-1] to obtain 2 (4.6 mg). The EtOAc residue (12 g) was subjected to silica gel column chromatography (220 g) and eluted with petroleumMe2CO mixtures in a gradient (1 : 0 → 0 : 1) to yield 11 fractions (1−11). Fraction 6 was purified by Sephadex LH-20 (MeOH), followed by RP-HPLC [MeOH/H2O (50 : 50), 4.0 mL·min−1] to obtain 1 (3.2 mg).
Structural Identification
Table 1 1H NMR and 13C NMR spectra for alkaloid 1 ( 600 MHz and 150 MHz, DMSO-d6) Position 1 2 3 4 4a 4b 5 6 7 8 8a 9 10 10a C=O NH 2-OCH3 4-OCH3 3-OH 6-OH
δH mult. (J Hz)
8.52, d (2.5) 7.04,dd (8.6, 2.5) 7.75,d (8.6) 7.05, s
10.73, s 4.33, s 4.01, s 9.36, s 9.65, s
δC
COSY
HMBC
H-7
C-4a, 4b, 6, 7, 8a
116.8
H-8
C-5, 8a
130.2 127.5 104.6 132.4 122.2 166.9
H-7
C-6, 8a, 9
109.9 148.8 143.8 150.1 127.1 115.8 111.0 155.8
62.7 60.0
C-8a, 10a
C-1, 10, 10a C-2 C-4
Enterocarpam-II (2) Brownish yellow needles (MeOH). 1H NMR (600 MHz, DMSO-d6) δ: 10.77 (1H, s, NH), 10.10 (1H, s, 3-OH), 8.62 (1H, d, J = 8.02 Hz, H-5), 7.85 (1H, s, H-9), 7.42 (1H, s, H-2) ,7.36 (1H, t, J = 8.02 Hz, H-6), 7.08 (1H, d, J = 8.02, H-7), 4.05 (3H, s, 4-OMe), 4.01 (3H, s, 8-OMe); 13C NMR (150 MHz, DMSO-d6) δ:168.3 (C=O), 154.2 (C-3), 153.7 (C-8), 150.5 (C-4), 133.9 (C-10), 127.0 (C-8a), 125.7 (C-1), 123.9 (C-6), 123.3 (C-6), 121.6 (C-5), 120.2 (C-10a), 117.9 (C-4b), 112.1 (C-7), 109.9 (C-2), 98.7 (C-9), 59.8 (4-OMe), 56.9 (8-OMe). Alkaloid 2 was identified as 10-amino-3-hydroxy-4, 8-dimethoxyphenanthrene-1-carboxylic acid lactam by comparison of the spectral data with the literature[8]. Oldhamactam (3) Brownish yellow needles (MeOH), 1 H NMR (600 MHz, DMSO-d6) δ: 10.91 (1H, s, NH), 10.15 (1H, s, 8-OH), 8.58 (1H, d, J = 8.4 Hz, H-5), 7.53 (1H, s, H-9), 7.35 (1H, t, H-6), 7.03 (1H, d, J = 8.4 Hz, H-7), 4.39 (3H, s, H-2), 4.08 (3H, s, 4-OMe), 3.91 (3H, s, 3-OMe); 13C NMR (150 MHz, DMSO-d6) δ: 166.2 (C=O), 156.3 (C-8), 153.5 (C-2), 153.3 (C-4), 146.0 (C-3), 133.2 (C-10), 126.8 (C-6), 125.7 (C-8a), 125.0 (C-4a), 122.9 (C-10a), 116.9 (C-5), 115.7 (C-1), 110.7 (C-8), 109.6 (C-4b), 99.5 (C-9), 62.4 (2OMe), 61.3 (4-OMe), 60.7 (3-OMe). Alkaloid 3 was identified as 10-amino-8-hydroxy-2, 3, 4-trimethoxyphenanthrene -1-carboxylic acid lactam by comparison of the spectral data with the literature[9].
ZHOU Xiao-Lei, et al. /Chinese Journal of Natural Medicines 2013, 11(1): 81−83
Hz, H-7), 4.37 (3H, s, H-2), 4.10 (3H, s, 4-OMe ), 3.91 (3H, s, 3-OMe); 13C NMR (150 MHz, DMSO-d6) δ: 166.5 (C=O), 155.8 (C-2), 146.1 (C-3), 156.6(C-4), 153.7(C-6), 132.1 (C-10), 130.3 (C-8), 127.6 (C-8a), 126.9 (C-10a), 125.8 (C-4a), 117.0 (C-7), 115.6 (C-4b), 111.1 (C-1), 110.3 (C-5), 106.1 (C-9), 63.2 (2-OMe), 61.9 (3-OMe), 61.2 (4-OMe). Alkaloid 5 was identified as 10-amino-6-hydroxy-2,3,4trimethoxyphenanthrene-1-carbo-xylic acid lactam by comparison of the spectral data with the literature [11].
References Fig. 1 Structures of alkaloids 1-5
[1]
Fig. 2 Key HMBC correlations of alkaloiud 2
Goniopedaline (4) Brownish yellow needles (MeOH), H NMR (600 MHz, DMSO-d6) δ: 10.90 (1H, s, NH), 9.52 (1H, s, 3-OH), 9.10 (1H, d, J = 7.38 Hz, H-5), 7.95 (1H, d, J = 7.38 Hz, H-8), 7.54 (2H, m, H-6, 7), 7.17 (1H, s, H-9), 4.36 (3H, s, H-2), 4.04 (3H, s, 4-OMe); 13C NMR (150 MHz, DMSO-d6) δ:166.6 (C=O), 149.6 (C-2), 148.5 (C-3), 143.9(C-4), 134.7 (C-10), 133.6 (C-4a), 128.7 (C-8), 126.3 (C-5), 125.9 (C-7), 125.6 (C-6), 122.0 (C-8a), 121.3 (C-10a), 115.7 (C-4b), 109.2 (C-1), 103.7 (C-9), 62.3 (2-OMe), 59.5 (4-OMe). Alkaloid 4 was identified as 10-amino-3-hydroxy-2, 4-dimethoxyphenanthrene-1-carboxylic acid lactam by comparison of the spectral data with the literature [10]. Stigmalactam (5) Brownish yellow needles (MeOH), 1 H NMR (600 MHz, DMSO-d6 ) δ:10.83 (1H, s, NH), 9.72 (1H, s, 6-OH), 8.52 (1H, d, J = 2.46 Hz, H-5), 7.79 (1H, d, J = 8.7 Hz, H-8), 7.15(1H, s, H-9), 7.06 (1H, dd, J = 8.7, 2.46 1
Jiang Y, Li PT. Flora of China [M]. Beijing: Science Press, 1979, 30(2): 164. [2] Liu YL, Ho DK, Cassady JM, et al. Dasytrichone, a novel flavone from Dasymaschalon trichophorum with cancer chemopreventive potential [J]. Nat Prod Res, 1992, 1(3): 161-165. [3] Andre S, Rudolf M, Thawatchai S, et al. Flavonol glycosides from Dasymaschalon sootepense [J]. Phytochemistry, 1998, 47(7): 1393-1396. [4] Andre S, Rudolf M, Ludger W, et al. Alkaloids from Dasymaschalon sootepense [J]. Biochem Syst Ecol, 1998, 26: 933-934. [5] Waraporn C, Patoomratana T, Natthinee A, et al. Cytotoxic alkaloids from stems, leaves and twigs of Dasymaschalon blumei [J]. Fitoterapia, 2011, 82: 964-968. [6] Chen YC, Chen JJ, Chang YL, et al. A new aristolactam alkaloids and anti-platelet aggregation constituents from Piper taiwanense [J]. Planta Med, 2004, 70: 174-177. [7] Wu TS, Leu YL, Chan YY. Constituents of the leaves of Aristolochia kaempferi [J]. Chem Pharm Bull, 1998, 46(10): 1624-1626. [8] Kamaliah M, Kai CC, Myung HP, et al. Aristolactams of Orophea enterocarpa [J]. Phytochemistry, 1986, 25(4): 965-967. [9] Zhang YN, Zhong XG, Zheng ZP, et al. Discovery and synthesis of new immunosuppressive alkaloids from the stem of Fissistigma oldhamii (Hemsl.) Merr. [J]. Bioorg Med Chem, 2007, 15: 988-996. [10] Yang XN, Jin YS, Zhu P, et al. Amides from Uvaria microcarpa [J]. Chem Nat Compd, 2010, 46(2): 324-326. [11] Chia YC, Chang FR, Teng CM, et al. Aristolactams and dioxoaporphines from Fissistigma balansae and Fissistigma oldhamii [J]. J Nat Prod, 2000, 63: 1160-1163.
皂帽花中的一个新生物碱 周晓磊 1, 2, 吴久鸿 1*, 白
皎 2, 胡晓兰 2，厉恩振 1, 3，史
宁 1，裴月湖 2
1
解放军第 306 医院药学部, 北京 100101; 沈阳药科大学中药学院, 沈阳 110016; 3 军事医学科学院毒物药物研究所, 北京 100850 2
【摘 要】 目的：对皂帽花化学成分的研究。方法：运用柱层析等方法分离纯化化合物, 通过波谱解析和理化鉴别进行结 构鉴定。结果：从皂帽花中分离得到 5 个生物碱，分别是：10-amino-3, 6-dihydroxy-2, 4-dimethoxyphenanthrene-1-carboxylic acid lactam (1), enterocarpam-II(2), oldhamactam (3), goniopedaline (4) 和 stigmalactam (5) 。结论：化合物 1 为新化合物, 化合物 2~5 均为首次从该属植物中分离得到。 【关键词】 番荔枝科; 皂帽花; 生物碱 【基金项目】
国家自然科学基金(No. 81072546)和全军“十一五”杰出人才基金（No. 06J001）资助项目
Chinese Journal of Natural Medicines
A new aristolactam alkaloid from the stems of Dasymaschalon trichophorum ZHOU Xiao-Lei1, 2, WU Jiu-Hong1*, BAI Jiao2, HU Xiao-Lan2, LI En-Zhen1, 3, SHI Ning1, PEI Yue-Hu2 1
Department of Pharmacy, The 306th hospital of PLA, Beijing 100101, China; School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang 110016, China; 3 Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Beijing 100850, China 2
Available online 20 Jan. 2013 [ABSTRACT] AIM: To study the chemical constituents of the stem of Dasymaschaon trichophorum Merr.. METHODS: The compounds were isolated and purified by the methods of various chromatography. Their structures were elucidated on the basis of spectroscopic methods. RESULTS: Five aristolactam alkaloids were obtained and their structures were identified as 10-amino-3,6-dihydroxy-2,4-dimethoxyphenanthrene-1-carboxylic acid lactam (1), enterocarpam-II (2), oldhamactam (3), goniopedaline (4), and stigmalactam (5), respectively. CONCLUSION: Compound 1 is a new aristolactam alkaloid, while compounds 2−5 are reported from this genus for the first time. [KEY WORDS] Annonaceae; Dasymaschaon trichophorum; Aristolactam alkaloids
[CLC Number] R284.1
1
[Document code] A
[Article ID] 1672-3651(2013)01-0081-03
Introduction
The genus Dasymaschalon Dalla Torre & Harms comprises about 16 species, three of which are distributed in the south of China[1]. To date, only a few chemical constituents of this genus have been reported, and most of them were flavonoids and alkaloids[2-5]. In a phytochemical study on the specie Dasymaschalon trichophorum Merr., five aristolactam alkaloids were isolated and their structures were elucidated on the basis of spectroscopic analysis as 10-amino-3, 6-dihydroxy-2, 4-dimethoxyphenanthrene-1-carboxylic acid lactam (1), a new alkaloid, and four known aristolactam alkaloids, enterocarpam-II (2), oldhamactam (3), goniopedaline (4), and stigmalactam (5).
2
Results and Discussion
Alkaloid 1 was isolated as brownish yellow needles. The molecular formula C17H13O5N was determined by
[Received on] 11-Apr.-2012 [Research funding] This project was supported by the National Natural Science Foundation of China (No.81072546) and the Chinese PLA National Science Fund for Distinguished Young Scholars Grant during the 11th “Five-Year” Plan period (No.06J001). [*Corresponding author] WU Jiu-Hong: Prof., Tel: 86-1066354564; Fax: 86-10-64879825, Email: [email protected] These authors have no conflict of interest to declare. Copyright © 2013, China Pharmaceutical University. Published by Elsevier B.V. All rights reserved
HR-ESI-MS at m/z 312.0867 [M + H]+ (Calcd. for C17H14O5 N1 312.086 6 ). The 13C NMR spectrum of alkaloid 1 indicated the presence of 15 sp2 carbons (including a carbonyl carbon at δ 166.9), which are typical for an aristolactam alkaloid[5-7]. Therefore, it was determined that 1 possesses an aristolactam skeleton. Comparing its 1H and 13C NMR data (see Table 1) with those of the known alkaloid goniopedaline (4), 1 had an additional hydroxyl group. In the 1H-NMR spectrum of 1, an ABX system for aromatic protons was observed at δ 8.52 (1H, d, J = 2.5 Hz), 7.75 (1H, d, J = 8.6 Hz), and 7.04 (1H, dd, J = 8.6, 2.5 Hz), attributable to H-5, H-8, and H-7, respectively. A singlet at δ 7.05 (1H) was assigned to H-9. These data resembled those of 5, 7, 8, 9-unsubstituted aristolactams. The HMBC correlations (see Table 1) between H-5 to C-6 and H-8 to C-6 confirmed the assignment of one hydroxyl OH (δ 9.65) at C-6. The remaining two methoxyl signals at δ 4.01 (3H, s) and 4.33 (3H, s) were assigned to C-4 and C-2. The phenolic hydroxyl (δ 9.36) could be assigned to C-3. These assignments were verified by HMBC correlations of the methoxyl protons at δ 4.01 with δ 150.1 (C-2), and δ 4.33 with δ 148.8 (C-4). The singlet signal at δ 10.73 was attributed to the amide NH, and this was confirmed by HMBC correlations between δ 10.73 to δ 109.9 (C-1), δ 132.4 (C-10) and δ 122.2 (C-10a). On the basis of these results, 1 was established structurally as 10-amino-3, 6dihydroxy-2, 4-dimethoxyphenanthrene-1-carb-oxylic acid lactam The structures of the known alkaloids (2−5) were identified by comparing their spectroscopic data (1H NMR and 13C
ZHOU Xiao-Lei, et al. /Chinese Journal of Natural Medicines 2013, 11(1): 81−83
NMR) with those reported in the literature.
4
3
10-Amino-3, 6-dihydroxy-2, 4- dimethoxyphenanthrene-1-carboxylic acid lactam (1) Brownish yellow needles (MeOH), HR-ESI-MS m/z 312.086 7 [M + H]+ (Calcd. for C17H14O5 N1, 312.086 6 ); 1H and 13C NMR data are shown in Table 1.
Experimental
3.1 General experimental procedures HR-ESI-MS was recorded on a Varian QFT-ESI mass spectrometer. 1H NMR and 13C NMR spectra were recorded on Bruker AV-600 NMR spectrometers using TMS as an internal standard. The chromatographic silica gel (54−74 μm) was purchased from Qingdao Marine Chemical Factory (Qingdao, China). Sephadex LH-20 was purchased from GE Healthcare. RP-HPLC analysis and semi-preparation were conducted using Hitachi L-6000 pumping system equipped with a Hitachi L-7400 UV detector and performed with a C18 column (10 mm × 250 mm, GL science Co., Ltd., Japan). Fractions were monitored by thin layer chromatography (TLC), and spots were detected with a UV 254 nm lamp and by spraying with 10% H2SO4 in EtOH followed by heating at 105 ºC for 5 min. 3.2 Plant material The stems of Dasymaschaon trichophorum Merr. were collected in Dec. 2010 in Beihai, Guangxi Zhuang Autonomous Region, China, and identified by Prof. FANG Ding (Guangxi Institute of Chinese Medicine & Pharmaceutical Sciences). A voucher specimen (No. 76773) was deposited in The 306th Hospital of PLA. 3.3 Extraction and isolation The dried and powered plant material (8.1 kg) was extracted with 95% EtOH three times under reflux. After concentration, the extract (750 g) was suspended in water and partitioned with CHCl3 followed by EtOAc. The CHCl3 residue (15 g) was subjected to silica gel column chromatography (600 g) and eluted with petroleum−EtOAc mixtures in a gradient (1 : 0 →0 : 1) to yield 15 fractions (1−15) . Fraction 10 was then subjected to column chromatography over silica gel (90 g) eluted with CHCl3−MeOH (100 : 1 →100 : 5) to give subfractions (Fr. A1−Fr. A5). Fr. A2 was purified by Sephadex LH-20 (MeOH), followed by RP-HPLC [MeOH/H2O (58 : 42), 4.0 mL·min−1] to obtain 4 (2.3 mg). Fr. A3 was further separated by silica gel chromatography, followed by Sephadex LH-20 column chromatography to obtain 3 (12.5 mg). Fr. A4 was purified by Sephadex LH-20 (MeOH), followed by RP-HPLC [MeOH/H2O (54 : 46), 4.0 mL·min−1] to obtain 5 (3.7 mg). Fraction 11 was subjected to column chromatography over silica gel (55 g) eluted with CHCl3−MeOH (9 : 2) to give subfractions (Fr. B1-Fr. B4). Fr. B4 was purified by Sephadex LH-20 (MeOH), followed by RP-HPLC [MeOH/H2O (52: 48), 4.0 mL·min-1] to obtain 2 (4.6 mg). The EtOAc residue (12 g) was subjected to silica gel column chromatography (220 g) and eluted with petroleumMe2CO mixtures in a gradient (1 : 0 → 0 : 1) to yield 11 fractions (1−11). Fraction 6 was purified by Sephadex LH-20 (MeOH), followed by RP-HPLC [MeOH/H2O (50 : 50), 4.0 mL·min−1] to obtain 1 (3.2 mg).
Structural Identification
Table 1 1H NMR and 13C NMR spectra for alkaloid 1 ( 600 MHz and 150 MHz, DMSO-d6) Position 1 2 3 4 4a 4b 5 6 7 8 8a 9 10 10a C=O NH 2-OCH3 4-OCH3 3-OH 6-OH
δH mult. (J Hz)
8.52, d (2.5) 7.04,dd (8.6, 2.5) 7.75,d (8.6) 7.05, s
10.73, s 4.33, s 4.01, s 9.36, s 9.65, s
δC
COSY
HMBC
H-7
C-4a, 4b, 6, 7, 8a
116.8
H-8
C-5, 8a
130.2 127.5 104.6 132.4 122.2 166.9
H-7
C-6, 8a, 9
109.9 148.8 143.8 150.1 127.1 115.8 111.0 155.8
62.7 60.0
C-8a, 10a
C-1, 10, 10a C-2 C-4
Enterocarpam-II (2) Brownish yellow needles (MeOH). 1H NMR (600 MHz, DMSO-d6) δ: 10.77 (1H, s, NH), 10.10 (1H, s, 3-OH), 8.62 (1H, d, J = 8.02 Hz, H-5), 7.85 (1H, s, H-9), 7.42 (1H, s, H-2) ,7.36 (1H, t, J = 8.02 Hz, H-6), 7.08 (1H, d, J = 8.02, H-7), 4.05 (3H, s, 4-OMe), 4.01 (3H, s, 8-OMe); 13C NMR (150 MHz, DMSO-d6) δ:168.3 (C=O), 154.2 (C-3), 153.7 (C-8), 150.5 (C-4), 133.9 (C-10), 127.0 (C-8a), 125.7 (C-1), 123.9 (C-6), 123.3 (C-6), 121.6 (C-5), 120.2 (C-10a), 117.9 (C-4b), 112.1 (C-7), 109.9 (C-2), 98.7 (C-9), 59.8 (4-OMe), 56.9 (8-OMe). Alkaloid 2 was identified as 10-amino-3-hydroxy-4, 8-dimethoxyphenanthrene-1-carboxylic acid lactam by comparison of the spectral data with the literature[8]. Oldhamactam (3) Brownish yellow needles (MeOH), 1 H NMR (600 MHz, DMSO-d6) δ: 10.91 (1H, s, NH), 10.15 (1H, s, 8-OH), 8.58 (1H, d, J = 8.4 Hz, H-5), 7.53 (1H, s, H-9), 7.35 (1H, t, H-6), 7.03 (1H, d, J = 8.4 Hz, H-7), 4.39 (3H, s, H-2), 4.08 (3H, s, 4-OMe), 3.91 (3H, s, 3-OMe); 13C NMR (150 MHz, DMSO-d6) δ: 166.2 (C=O), 156.3 (C-8), 153.5 (C-2), 153.3 (C-4), 146.0 (C-3), 133.2 (C-10), 126.8 (C-6), 125.7 (C-8a), 125.0 (C-4a), 122.9 (C-10a), 116.9 (C-5), 115.7 (C-1), 110.7 (C-8), 109.6 (C-4b), 99.5 (C-9), 62.4 (2OMe), 61.3 (4-OMe), 60.7 (3-OMe). Alkaloid 3 was identified as 10-amino-8-hydroxy-2, 3, 4-trimethoxyphenanthrene -1-carboxylic acid lactam by comparison of the spectral data with the literature[9].
ZHOU Xiao-Lei, et al. /Chinese Journal of Natural Medicines 2013, 11(1): 81−83
Hz, H-7), 4.37 (3H, s, H-2), 4.10 (3H, s, 4-OMe ), 3.91 (3H, s, 3-OMe); 13C NMR (150 MHz, DMSO-d6) δ: 166.5 (C=O), 155.8 (C-2), 146.1 (C-3), 156.6(C-4), 153.7(C-6), 132.1 (C-10), 130.3 (C-8), 127.6 (C-8a), 126.9 (C-10a), 125.8 (C-4a), 117.0 (C-7), 115.6 (C-4b), 111.1 (C-1), 110.3 (C-5), 106.1 (C-9), 63.2 (2-OMe), 61.9 (3-OMe), 61.2 (4-OMe). Alkaloid 5 was identified as 10-amino-6-hydroxy-2,3,4trimethoxyphenanthrene-1-carbo-xylic acid lactam by comparison of the spectral data with the literature [11].
References Fig. 1 Structures of alkaloids 1-5
[1]
Fig. 2 Key HMBC correlations of alkaloiud 2
Goniopedaline (4) Brownish yellow needles (MeOH), H NMR (600 MHz, DMSO-d6) δ: 10.90 (1H, s, NH), 9.52 (1H, s, 3-OH), 9.10 (1H, d, J = 7.38 Hz, H-5), 7.95 (1H, d, J = 7.38 Hz, H-8), 7.54 (2H, m, H-6, 7), 7.17 (1H, s, H-9), 4.36 (3H, s, H-2), 4.04 (3H, s, 4-OMe); 13C NMR (150 MHz, DMSO-d6) δ:166.6 (C=O), 149.6 (C-2), 148.5 (C-3), 143.9(C-4), 134.7 (C-10), 133.6 (C-4a), 128.7 (C-8), 126.3 (C-5), 125.9 (C-7), 125.6 (C-6), 122.0 (C-8a), 121.3 (C-10a), 115.7 (C-4b), 109.2 (C-1), 103.7 (C-9), 62.3 (2-OMe), 59.5 (4-OMe). Alkaloid 4 was identified as 10-amino-3-hydroxy-2, 4-dimethoxyphenanthrene-1-carboxylic acid lactam by comparison of the spectral data with the literature [10]. Stigmalactam (5) Brownish yellow needles (MeOH), 1 H NMR (600 MHz, DMSO-d6 ) δ:10.83 (1H, s, NH), 9.72 (1H, s, 6-OH), 8.52 (1H, d, J = 2.46 Hz, H-5), 7.79 (1H, d, J = 8.7 Hz, H-8), 7.15(1H, s, H-9), 7.06 (1H, dd, J = 8.7, 2.46 1
Jiang Y, Li PT. Flora of China [M]. Beijing: Science Press, 1979, 30(2): 164. [2] Liu YL, Ho DK, Cassady JM, et al. Dasytrichone, a novel flavone from Dasymaschalon trichophorum with cancer chemopreventive potential [J]. Nat Prod Res, 1992, 1(3): 161-165. [3] Andre S, Rudolf M, Thawatchai S, et al. Flavonol glycosides from Dasymaschalon sootepense [J]. Phytochemistry, 1998, 47(7): 1393-1396. [4] Andre S, Rudolf M, Ludger W, et al. Alkaloids from Dasymaschalon sootepense [J]. Biochem Syst Ecol, 1998, 26: 933-934. [5] Waraporn C, Patoomratana T, Natthinee A, et al. Cytotoxic alkaloids from stems, leaves and twigs of Dasymaschalon blumei [J]. Fitoterapia, 2011, 82: 964-968. [6] Chen YC, Chen JJ, Chang YL, et al. A new aristolactam alkaloids and anti-platelet aggregation constituents from Piper taiwanense [J]. Planta Med, 2004, 70: 174-177. [7] Wu TS, Leu YL, Chan YY. Constituents of the leaves of Aristolochia kaempferi [J]. Chem Pharm Bull, 1998, 46(10): 1624-1626. [8] Kamaliah M, Kai CC, Myung HP, et al. Aristolactams of Orophea enterocarpa [J]. Phytochemistry, 1986, 25(4): 965-967. [9] Zhang YN, Zhong XG, Zheng ZP, et al. Discovery and synthesis of new immunosuppressive alkaloids from the stem of Fissistigma oldhamii (Hemsl.) Merr. [J]. Bioorg Med Chem, 2007, 15: 988-996. [10] Yang XN, Jin YS, Zhu P, et al. Amides from Uvaria microcarpa [J]. Chem Nat Compd, 2010, 46(2): 324-326. [11] Chia YC, Chang FR, Teng CM, et al. Aristolactams and dioxoaporphines from Fissistigma balansae and Fissistigma oldhamii [J]. J Nat Prod, 2000, 63: 1160-1163.
皂帽花中的一个新生物碱 周晓磊 1, 2, 吴久鸿 1*, 白
皎 2, 胡晓兰 2，厉恩振 1, 3，史
宁 1，裴月湖 2
1
解放军第 306 医院药学部, 北京 100101; 沈阳药科大学中药学院, 沈阳 110016; 3 军事医学科学院毒物药物研究所, 北京 100850 2
【摘 要】 目的：对皂帽花化学成分的研究。方法：运用柱层析等方法分离纯化化合物, 通过波谱解析和理化鉴别进行结 构鉴定。结果：从皂帽花中分离得到 5 个生物碱，分别是：10-amino-3, 6-dihydroxy-2, 4-dimethoxyphenanthrene-1-carboxylic acid lactam (1), enterocarpam-II(2), oldhamactam (3), goniopedaline (4) 和 stigmalactam (5) 。结论：化合物 1 为新化合物, 化合物 2~5 均为首次从该属植物中分离得到。 【关键词】 番荔枝科; 皂帽花; 生物碱 【基金项目】
国家自然科学基金(No. 81072546)和全军“十一五”杰出人才基金（No. 06J001）资助项目