中草药  2016, Vol. 47 Issue (8): 1278-1281
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引用本文doi: 10.7501/j.issn.0253-2670.2016.08.005
李宛霏, 邹娟, 李继新, 张敬杰, 潘炉台. 岩凤尾蕨化学成分的研究[J]. 中草药, 2016, 47(8): 1278-1281.
LI Wan-fei, ZOU Juan, LI Ji-xin, ZHANG Jing-jie, PAN Lu-tai. Chemical constituents from Pteris deltodon[J]. Chinese Traditional and Herbal Drugs, 2016, 47(8): 1278-1281.
岩凤尾蕨化学成分的研究
李宛霏, 邹娟, 李继新, 张敬杰 , 潘炉台    
贵阳中医学院, 贵州 贵阳 550025
收稿日期: 2015-12-31;
基金项目: 国家自然科学基金资助项目(81160496);贵州省科学技术基金项目(黔科合J字 [2013] 2070号)
作者简介: 李宛霏(1989—),女,硕士研究生,从事天然药物化学研究。E-mail:wanfeier@live.cn
通讯作者: 张敬杰(1971—),女,教授,硕士生导师,从事民族药资源及化学的研究。Tel:(0851)88233016 E-mail:icezjj@tom.com
摘要: 目的 研究岩凤尾蕨Pteris deltodon全草的化学成分。方法 使用硅胶柱色谱、Sephadex LH-20及半制备高效液相色谱法等手段进行分离纯化,通过光谱数据结合参考文献鉴定化合物结构。结果 从岩凤尾蕨全草95%乙醇提取部位分离到9个化合物,分别鉴定为β-谷甾醇(1)、过氧麦角甾醇(2)、2β,15α-二羟基-对映-贝壳杉-16-烯(3)、3β,25-二羟基-环阿尔廷-23-烯(4)、3β,24-二羟基-环阿尔廷-25-烯(5)、大黄素(6)、1,7-二羟基 酮(7)、熊果酸(8)、麦角甾醇(9)。结论 所有化合物均首次从该植物中分离到,化合物4和5为首次从凤尾蕨属植物分离到的环阿尔廷烷型三萜。
关键词: 岩凤尾蕨     环阿尔廷烷三萜     3β,24-二羟基-环阿尔廷-25-烯     大黄素     熊果酸    
Chemical constituents from Pteris deltodon
LI Wan-fei , ZOU Juan, LI Ji-xin, ZHANG Jing-jie, PAN Lu-tai    
Guiyang College of Traditional Chinese Medicine, Guiyang 550025, China
Abstract: Objective To study the chemical constituents in the whole plant of Pteris deltodon. Methods Nine compounds were isolated from 95% ethanol extract in the whole plant of P. deltodon and purified by silica gel chromatography, Sephadex LH-20, and pre-HPLC, and their structures were identified on the basis of spectroscopic data and literatures. Results Nine compounds were isolated and elucidated as β-sitosteol (1), (22E)-5α,8α-epidioxyergosta-6,22-dien-3β-ol (2), ent-kaur-16-ene-2β,15α-diol (3), cycloart-23-en-3β,25-diol (4), cycloart-25-en-3β,24-diol (5), emodin (6), 1,7-dihydroxyxanthone (7), ursolic acid (8), and ergosterol (9). Conclusion All the compounds are for the first time isolated from P. deltodon; Compound 4 and 5 are cycloartanes first isolated from genus Pteris L.
Key words: Pteris deltodon Bak.     cycloartane triterpene alkyl     cycloart-25-en-3β,24-diol     emodin     ursolic acid    

岩凤尾蕨Pteris deltodon Bak. 系凤尾蕨科（Pteridaceae）凤尾蕨属Pteris L. 植物，主要分布于云南、四川、广西、贵州、广东等地[1]。岩凤尾蕨全草入药；味甘、苦，性凉；清热利湿、敛肺止咳、定惊、解毒；主治泄泻、痢疾、久咳不止、小儿惊风等[2]。近年来，本课题组从蕨类植物资源中寻找活性成分，先后研究了长柄石杉、雷山石杉等石杉科植物的化学成分[3]。凤尾蕨属植物中因含有5F（11β-羟基-对映-贝壳杉-16-烯-15-酮-19-酸）和6F（7β,9α-二羟基-对映-贝壳杉-16-烯-15-酮-19,6α-内酯）等抗肿瘤活性的二萜类化合物[4, 5]，引起人们关注，为寻找结构新颖又具活性的先导化合物，本实验对岩凤尾蕨进行研究，从该植物全草中分离得到了9个化合物，分别鉴定为β-谷甾醇（β-sitosteol，1）、过氧麦角甾醇 [(22E)-5α,8α-epidioxyergosta- 6,22-dien-3β-ol，2]、2β,15α-二羟基-对映-贝壳杉-16-烯（ent-kaur-16-ene-2β,15α-diol，3）、3β,25-二羟基-环阿尔廷-23-烯（cycloart-23-en-3β,25-diol，4）、3β,24-二羟基-环阿尔廷-25-烯（cycloart-25-en-3β,24-diol，5）、大黄素（emodin，6）、1,7-二羟基𠮿酮（1,7-dihydroxyxanthone，7）、熊果酸（ursolic acid，8）、麦角甾醇（ergosterol，9）。化合物1～9均为首次从该植物中分离到；化合物4和5为首次从凤尾蕨属植物中分离得到的环阿尔廷烷型三萜化合物。

1 仪器和材料

JEOL 5973 MSD质谱仪（安捷伦公司）；ECX-1MHz、JEOL-400 MHz超导核磁共振谱仪（日本电子株式会社）；分析和半制备型高效液相色谱仪为Agilent 1100；R-201型旋转蒸发仪（上海申胜生物技术有限公司）HP8453；WFH-308B三色紫外分析仪（上海精科实业有限公司）；Metter-Toledo电子天平（瑞士Metter-Toledo公司）；Sephadex LH-20（瑞士Pharmacia＆Upjohn公司）；MCI（75～150 m）为日本Mitsubishi化学公司生产；制备薄层色谱（GF254）及柱色谱（200～300目）用硅胶（青岛海洋化工有限公司）；其余试剂均为重蒸工业纯。

实验药材于2013年7月采自贵州省大方县，原植物经贵阳中医学院赵俊华教授鉴定为凤尾蕨科凤尾蕨属植物岩凤尾蕨Pteris deltodon Bak.，凭证标本（GY20130706）存于贵阳中医学院。

2 提取与分离

取干燥的岩凤尾蕨全草粉末15 kg，用95%的乙醇回流提取3次，每次3 h，合并提取液并减压浓缩得浸膏300 g。经硅胶柱梯度洗脱（石油醚-醋酸乙酯100∶1→6∶4）得流分Fr. A～E。

Fr. A经硅柱石油醚-醋酸乙酯（100∶1→1∶1）系统梯度洗脱，在石油醚-醋酸乙酯9∶1部位减压浓缩后得到化合物1（100 mg）。Fr. C和D经MCI脱色后的流分通过硅胶色谱柱石油醚-醋酸乙酯（9∶1→7∶3）梯度洗脱，石油醚-醋酸乙酯8∶2部分继续进行硅胶柱色谱得到化合物2（55 mg）。石油醚-醋酸乙酯7∶3部分减压浓缩后得到粉末4（10 mg），母液经Sephadex LH-20（氯仿-甲醇，1∶1）洗脱得化合物5（10 mg）。石油醚-醋酸乙酯9∶1部分通过硅胶色谱柱石油醚-醋酸乙酯（10∶1→9∶1）梯度洗脱，在石油醚-醋酸乙酯10∶1部分得到化合物 6（8 mg）。Fr. E经MCI脱色得到化合物3（70 mg）。脱色后通过硅胶柱色谱，石油醚-醋酸乙酯（10∶1→7∶3）梯度洗脱，得到化合物7（20 mg）；母液经硅胶柱色谱分离，得到化合物8（10 mg）和9（169 mg）。

3 结构鉴定

化合物1：白色针晶（甲醇），mp 134～136 ℃，分子式为C29H50O，ESI-MS m/z: 415 [M＋H]+。1H-NMR (400 MHz,CDCl3) δ: 0.68 (3H,s,H-18),0.82 (3H,d,J = 7.7 Hz,H-27),0.85 (3H,brd,J = 7.7 Hz,H-26),0.90 (3H,brt,J = 9.3 Hz,H-29),0.94 (3H,brd,J = 8.3 Hz,H-21),1.00 (3H,s,H-19),3.50 (1H,m,H-3),5.35 (1H,brd,J = 5.2 Hz,H-6)；13C-NMR (100 MHz,CDCl3) δ: 37.7 (C-1),32.1 (C-2),72.3 (C-3),42.8 (C-4),141.2 (C-5),122.2 (C-6),32.4 (C-7),32.1 (C-8),50.6 (C-9),37.0 (C-10),21.5 (C-11),40.2 (C-12),42.9 (C-13),57.2 (C-14),24.8 (C-15),28.7 (C-16),56.5 (C-17),12.3 (C-18),20.3 (C-19),37.0 (C-20),19.3 (C-21),34.4 (C-22),26.5 (C-23),46.3 (C-24),29.6 (C-25),20.3 (C-26),19.9 (C-27),23.5 (C-28),12.5 (C-29)。以上数据与文献报道基本一致[6]，故鉴定化合物1为β-谷甾醇。

化合物2：淡黄色粉末，mp 177～178 ℃，分子式为C28H44O3，ESI-MS m/z: 429 [M＋H]+。1H-NMR (400 MHz,CDCl3) δ: 0.80 (3H,d,J = 6.8 Hz,H-26),0.82 (3H,s,H-18),0.83 (3H,d,J = 6.8 Hz,H-27),0.88 (3H,s,H-19),0.91 (3H,s,H-28),1.00 (3H,d,J = 6.8 Hz,H-21),3.98 (1H,m,H-3),5.15 (1H,dd,J = 15.3,8.1 Hz,H-22),5.21 (1H,dd,J = 15.3,7.4 Hz,H-23),6.24 (1H,d,J = 8.5 Hz,H-6),6.51 (1H,d,J = 8.5 Hz,H-7)；13C-NMR (100 MHz,CDCl3) δ: 34.8 (C-1),29.9 (C-2),66.6 (C-3),37.1 (C-4),82.3 (C-5),135.5 (C-6),130.9 (C-7),79.6 (C-8),51.2 (C-9),37.0 (C-10),23.5 (C-11),39.5 (C-12),44.7 (C-13),51.8 (C-14),20.8 (C-15),28.8 (C-16),56.3 (C-17),13.0 (C-18),18.3 (C-19),39.9 (C-20),21.0 (C-21),135.3 (C-22),132.4 (C-23),42.9 (C-24),33.2 (C-25),19.8 (C-26),20.1 (C-27),17.7 (C-28)。以上数据与文献报道基本一致[7]，故 鉴定化合物2为过氧麦角甾醇。

化合物3：白色粉末，mp 218～219 ℃，分子式为C20H32O2，ESI-MS m/z: 305 [M＋H]+。1H-NMR (400 MHz,CDCl3) δ: 0.87 (1H,dd,J = 11.0,0.9 Hz,H-5),0.93 (3H,s,H-19),0.97 (3H,s,H-18),1.11 (1H,m,H-9),1.09 (3H,s,H-20),1.54 (1H,m,H-13),3.84 (1H,s,H-15),3.95 (1H,m,H-2),5.09 (1H,s,H-17),5.22 (1H,s,H-17)；13C-NMR (100 MHz,CDCl3) δ: 50.0 (C-1),65.6 (C-2),51.6 (C-3),42.7 (C-4),56.0 (C-5),19.6 (C-6),19.2 (C-7),48.0 (C-8),54.5 (C-9),35.3 (C-10),36.9 (C-11),41.7 (C-12),33.1 (C-13),35.5 (C-14),83.4 (C-15),160.5 (C-16),108.9 (C-17),34.2 (C-18),22.9 (C-19),18.6 (C-20)。以上数据与文献报道基本一致[8]，故鉴定化合物3为2β,15α-二羟基-对映-贝壳杉-16-烯。

化合物4：白色粉末，mp 161～163 ℃，分子式为C30H50O2，ESI-MS m/z: 443 [M＋H]+。1H-NMR (400 MHz,C5D5N) δ: 0.80 (3H,s,H-29),0.84 (3H,d,J = 6.8 Hz,H-21),0.86 (3H,s,H-28),0.90 (6H,s,H-18,30),1.31 (6H,s,H-26,27),3.28 (1H,dd,J = 11.1,4.3 Hz,H-3),5.59 (2H,m,H-23,24)；13C-NMR (100 MHz,C5D5N) δ: 32.0 (C-1),30.4 (C-2),78.9 (C-3),40.5 (C-4),47.1 (C-5),21.2 (C-6),26.5 (C-7),48.1 (C-8),20.0 (C-9),26.1 (C-10),26.1 (C-11),32.8 (C-12),45.3 (C-13),48.9 (C-14),35.6 (C-15),28.1 (C-16),52.0 (C-17),18.2 (C-18),30.0 (C-19),36.4 (C-20),18.3 (C-21),39.1 (C-22),125.5 (C-23),139.4 (C-24),70.8 (C-25),29.9 (C-26),29.9 (C-27),19.4 (C-28),14.1 (C-29),25.5 (C-30)。以上数据与文献报道基本一致[9]，故鉴定化合物4为3β,25-二羟基-环阿尔廷-23-烯。

化合物5：白色粉末，mp 149～151 ℃，分子式为C30H50O2，ESI-MS m/z: 443 [M＋H]+。1H-NMR (400 MHz,CDCl3) δ: 0.80 (3H,s,H-29),0.85 (3H,d,J = 5.3 Hz,H-21),0.90 (3H,s,H-28),0.96 (6H,s,H-18,30),1.71 (3H,s,H-27),3.28 (1H,dd,J = 11.1,4.3 Hz,H-3),4.80 (1H,brs,H-26a),4.90 (1H,brs,H-26b)；13C-NMR (100 MHz,CDCl3) δ: 32.0 (C-1),30.1 (C-2),78.9 (C-3),40.5 (C-4),47.1 (C-5),21.1 (C-6),28.1 (C-7),48.0 (C-8),20.0 (C-9),26.0 (C-10),26.1 (C-11),32.9 (C-12),45.3 (C-13),48.8 (C-14),35.6 (C-15),26.5 (C-16),52.2 (C-17),18.0 (C-18),29.9 (C-19),36.0 (C-20),18.3 (C-21),32.0 (C-22),31.6 (C-23),76.4 (C-24),147.9 (C-25),110.9 (C-26),17.6 (C-27),19.3 (C-28),25.4 (C-29),14.0 (C-30)。以上数据与文献报道基本一致[10]，故鉴定化合物5为3β,24-二羟基-环阿尔廷-25-烯。

化合物6：橙色针晶（甲醇），mp 254～256 ℃，分子式为C15H10O5，ESI-MS m/z: 271 [M＋H]+。1H-NMR (400 MHz,CD3COCD3) δ: 12.15 (1H,s,1-OH),12.03 (1H,s,8-OH),10.38 (1H,s,3-OH),7.52 (1H,s,H-5),7.20 (1H,m,H-4),7.09 (1H,m,H-7),6.63 (1H,m,H-2),2.45 (3H,s,CH3)；13C-NMR (100 MHz,CD3COCD3) δ: 166.2 (C-1),108.8 (C-2),166.4 (C-3),109.7 (C-4),121.4 (C-5),149.5 (C-6),124.9 (C-7),163.2 (C-8),191.6 (C-9),182.1 (C-10),134.2 (C-11),114.4 (C-12),110.3 (C-13),136.5 (C-14),22.0 (CH3)。以上数据与文献报道基本一致[11]，故鉴定化合物6为大黄素。

化合物7：黄色针晶（甲醇），mp 241～242 ℃，分子式为C13H8O4，ESI-MS m/z: 229 [M＋H]+。1H-NMR (400 MHz,CD3OD) δ: 6.75 (1H,d,J = 8.2 Hz,H-2),6.94 (1H,d,J = 8.2 Hz,H-4),7.32 (1H,dd,J = 8.8,2.8 Hz,H-6),7.43 (1H,d,J = 8.8 Hz,H-5),7.50 (1H,d,J = 2.8 Hz,H-8),7.64 (1H,t,J = 8.2 Hz,H-3)；13C-NMR (100 MHz,CD3OD) δ: 162.9 (C-1),110.7 (C-2),137.9 (C-3),108.1 (C-4),120.2 (C-5),126.3 (C-6),155.5 (C-7),109.5 (C-8),183.3 (C-9),157.5 (C-4a),122.2 (C-8a),109.1 (C-9a),151.5 (C-10a)。以上数据与文献报道基本一致[12]，故鉴定化合物7为1,7-二羟基𠮿酮。

化合物8：白色粉末，mp 257～259 ℃，分子式为C30H48O3，ESI-MS m/z: 457 [M＋H]+。1H-NMR (400 MHz,C5D5N) δ: 0.90 (3H,s,H-24),0.97 (3H,d,J = 6.0 Hz,H-29),1.08 (6H,s,H-25,27),1.05 (3H,d,J = 6.6 Hz,H-30),1.25 (3H,s,H-23),1.27 (3H,s,H-26),2.66 (1H,d,J = 11.4 Hz,H-18),3.48 (1H,dd,J = 10.8,6.0 Hz,H-3),5.52 (1H,brs,H-12)；13C-NMR (100 MHz,C5D5N) δ: 39.2 (C-1),28.8 (C-2),78.3 (C-3),40.1 (C-4),55.9 (C-5),18.9 (C-6),33.7 (C-7),39.2 (C-8),48.2 (C-9),37.4 (C-10),17.7 (C-11),125.8 (C-12),139.4 (C-13),42.3 (C-14),29.0 (C-15),24.1 (C-16),48.2 (C-17),53.7 (C-18),39.2 (C-19),39.6 (C-20),31.2 (C-21),37.4 (C-22),29.0 (C-23),16.8 (C-24),15.9 (C-25),17.6 (C-26),24.1 (C-27),180.1 (C-28),24.1 (C-29),21.6 (C-30)。以上数据与文献报道基本一致[11]，故鉴定化合物8为熊果酸。

化合物9：白色针晶（甲醇）mp 158～160 ℃，分子式为C28H44O，ESI-MS m/z: 397 [M＋H]+。1H-NMR (400 MHz,CDCl3) δ: 0.68 (3H,s,H-18),0.84 (3H,d,J = 6.0 Hz,H-27),0.86 (3H,d,J = 7.2 Hz,H-26),0.90 (3H,d,J = 6.9 Hz,H-28),0.94 (3H,s,H-19),1.00 (3H,d,J = 6.5 Hz,H-21),3.50 (1H,m,H-3),5.17 (1H,dd,J = 15.3,8.3 Hz,H-22),5.20 (1H,dd,J = 15.3,7.4 Hz,H-23),5.35 (1H,brd,J = 5.2 Hz,H-6)；13C-NMR (100 MHz,CDCl3) δ: 38.3 (C-1),31.9 (C-2),70.4 (C-3),40.4 (C-4),141.3 (C-5),119.5 (C-6),116.2 (C-7),139.7 (C-8),46.2 (C-9),37.0 (C-10),21.1 (C-11),38.8 (C-12),42.8 (C-13),54.5 (C-14),22.9 (C-15),28.2 (C-16),55.7 (C-17),11.9 (C-18),16.2 (C-19),39.0 (C-20),19.0 (C-21),135.0 (C-22),131.9 (C-23),40.7 (C-24),33.0 (C-25),20.2 (C-26),20.0 (C-27),17.6 (C-28)。以上数据与文献报道一致[13]，故鉴定化合物9为麦角甾醇。

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