南蛇风化学成分的研究

陈庆淑 周小雷陈 路江 姗陈誉丹马国需刘华钢袁经权*

[1.广西中医药大学科学实验中心,广西 南宁 530200;2.海南省人民医院(海南医学院附属海南医院),海南 海口 570311;3.中国医学科学院药用植物研究所广西分所,广西 南宁 530023]

瑶药南蛇风亦称南蛇簕,为苏木科云实属植物喙荚云实Caesalpinia minax Hance 的藤茎,分布广西、广东、福建、台湾等省区,为我国瑶族地区104 种老班瑶药之一,主要用于哈轮(感冒)、泵烈竞(尿路感染)、月藏(尿血)、就港虾(急性肠胃炎)、碰累(痢疾) 等症,为瑶医常用打药之一[1]。国内外学者研究发现喙荚云实具有抗流感甲型病毒、抗肿瘤、抗带状疱疹、抗蛇毒、镇痛以及抗炎等药理作用[2-4]。本课题组前期从喙荚云实种子(苦石莲) 中分离到一系列新颖卡萨烷二萜成分,具有良好抗肿瘤活性等[5-8]。但国内外对瑶药南蛇风,即喙荚云实藤茎部位的化学成分鲜有报道。为进一步研究喙荚云实植物,开发瑶药,本实验初步对南蛇风进行化学成分研究,从其乙醇提取物中分离鉴定了11 个化合物,其中化合物2～3、5～6、9 为首次从该属植物中分离得到,化合物1、7 为首次从该植物中分离得到。

1 材料

Trace DSQ 四极杆型质谱仪(美国Finnigan 公司)；Dre-600 型核磁共振光谱仪(美国瓦里安公司)；薄层色谱和柱色谱硅胶(青岛海洋化工有限公司)。常规试剂均为分析纯。

南蛇风采自广西壮族自治区南宁市邕宁区三塘镇,经广西中医药大学药用植物教研室韦松基教授鉴定为苏木科云实属植物喙荚云实Caesalpinia minax Hance 的藤茎,标本保存于广西中医药大学科学实验中心。

2 提取与分离

将药材阴干后称取20 kg,粉碎,加入85%乙醇浸渍渗漉提取,收集合并渗漉液,减压回收乙醇至基本无乙醇味。总浸膏加入适量水进行分散均匀后,依次以石油醚、氯仿、乙酸乙酯、正丁醇进行萃取。取氯仿萃取物(90 g) 上硅胶色谱柱,分别在用石油醚-乙酸乙酯(100∶0～0∶100) 洗脱,在石油醚-乙酸乙酯(40∶1) 的洗脱部分得到2 个粗粉末,取其中一粉末经硅胶柱、结晶,得到化合物1 (30 mg)；另一粗粉末经石油醚反复洗除色素,重结晶,得到化合物2 (160 mg)。在石油醚-乙酸乙酯(10∶1) 的洗脱部分得到黄色胶状物和黄色粉末,黄色胶状物经硅胶柱、结晶、重结晶,得到化合物3 (250 mg)、10 (150 mg)；黄色粉末用氯仿-丙酮(20∶1～1∶1) 洗脱,在乙酸乙酯中析出化合物4 (40 mg)。在石油醚-乙酸乙酯(5∶1) 的洗脱部分得到白色针状结晶和白色粉末,白色针状结晶经硅胶柱、结晶,得到化合物5 (12 mg)；白色粉末通过石油醚-丙酮(10∶1～1∶1)洗脱,得到化合物6 (15 mg)。用石油醚-乙酸乙酯(1∶1) 洗脱,得到化合物7 (17 mg)。用乙酸乙酯洗脱,得到白色絮状物和白色粉末,白色絮状物经Sephadex LH-20 柱,得到化合物8 (7 mg)、9(8 mg),白色粉末用甲醇重复洗除色素,得到化合物11 (35 mg)。

3 结构鉴定

化合物1:白色针状结晶,ESI-MS m/z:427.5 [M+H]+。1H-NMR (CDCl3,600 MHz) δ:0.65,0.80,0.88,0.93,0.94,0.98,1.11 (each 3H,s,7×CH3),0.81 (3H,d,J=6.8 Hz,H-4),提示分子式含有8 个甲基,其中7 个为孤立甲基；13C-NMR (CDCl3,150 MHz) δ:21.3 (C-1),40.5 (C-2),212.1 (C-3),57.2 (C-4),41.1(C-5),40.3 (C-6),17.2 (C-7),52.1 (C-8),36.4 (C-9),58.5 (C-10),34.6 (C-11),29.5(C-12),38.7 (C-13),37.3 (C-14),31.4 (C-15),35.0 (C-16),29.0 (C-17),41.8 (C-18),34.3 (C-19),27.2 (C-20),31.8 (C-21),38.2(C-22),5.8 (C-23),13.6 (C-24),16.9 (C-25),19.2 (C-26),17.7 (C-27),31.1 (C-28),34.0 (C-29),30.8 (C-30)。以上数据与文献[9]基本一致,故鉴定为木栓酮。

化合物2:白色粉末,ESI-MS m/z:201.6[M+H]+。1H-NMR (CDCl3,600 MHz) δ:0.88(3H,m,-CH3)；13C-NMR (CDCl3,150 MHz) δ:63.5 (C-OH),32.8,31.9,29.6,29.6,29.6,29.6,29.4,29.3,25.7,24.7,22.6,14.0 (13个脂肪C)。以上数据与文献[10]基本一致,故鉴定为正十三醇。

化合物3:白色针状结晶,ESI-MS m/z:457.4 [M+H]+。1H-NMR (CD3OD,600 MHz) δ:4.69 (1H,brs,H-30α),4.57 (1H,brs,H-30β),3.33 (1H,H-3),0.74,0.84,0.93,0.95,0.99,1.68 (6×CH3)；13C-NMR (CD3OD,150 MHz) δ:38.5 (C-1),26.6 (C-2),77.8(C-3),38.6 (C-4),55.4 (C-5),18.0 (C-6),34.2 (C-7),40.5 (C-8),50.5 (C-9),36.9(C-10),20.7 (C-11),25.5 (C-12),38.2 (C-13),42.2 (C-14),30.3 (C-15),31.9 (C-16),56.0 (C-17),47.0 (C-18),48.4 (C-19),150.3 (C-20),29.4 (C-21),36.7 (C-22),27.1 (C-23),14.6 (C-24),15.3 (C-25),15.2(C-26),13.6 (C-27),178.5 (C-28),19.0 (C-29),108.6 (C-30)。以上数据与文献[11]基本一致,并且与白桦脂酸标准对照品通过薄层色谱进一步鉴定,在3 种不同的展开系统中比较色谱行为完全一致,故鉴定为白桦脂酸。

化合物4:黄色针状结晶,ESI-MS m/z:283.1 [M+H]+。1H-NMR (CD3OD,600 MHz) δ:5.34 (2H,d,J=1.9 Hz,H-2),7.79 (1H,d,J=8.5 Hz,H-5),6.52 (1H,dd,J=8.6,2.2 Hz,H-6),6.29 (1H,d,J=2.1 Hz,H-8),7.71 (1H,brs,H-9),7.33 (2H,d,J=8.1 Hz,H-2′,6′),7.02 (2H,d,J=8.5 Hz,H-3′,5′),3.84 (3H,s,-OCH3)；13C-NMR (CD3OD,150 MHz) δ:67.6 (C-2),128.9 (C-3),181.5(C-4),129.3 (C-5),110.8 (C-6),165.2 (C-7),102.2 (C-8),136.2 (C-9),163.4 (C-10),114.4 (C-11),126.8 (C-1′),131.8 (C-2′),113.9 (C-3′),160.9 (C-4′),113.9 (C-5′),131.8 (C-6′),54.5 (-OCH3)。以上数据与文献[12]基本一致,故鉴定为bonducellin。

化合物5:白色针状结晶,ESI-MS m/z:443.2 [M-H ]-,467.3 [M+Na ]+。1H-NMR(CDCl3,600 MHz) δ:7.70 (2H,d,J=7.4 Hz,H-16,20),7.51 (1H,t,J=7.1 Hz,H-18),7.41(2H,t,J=7.6 Hz,H-17,19),2.95 (3H,s,H-12)；13C-NMR (CDCl3,150 MHz) δ:170.4 (C-1,amide),49.6 (C-2),37.6 (C-3),136.8 (C-4),128.8 (C-5),129.0 (C-6),127.0 (C-7),129.0 (C-8),128.8 (C-9),64.8 (C-10),171.0(C-11,ester),21.0 (C-12),55.2 (C-13),167.3(C-14,amide),133.8 (C-15),127.2 (C-16),128.1 (C-17),132.1 (C-18),128.1 (C-19),127.2 (C-20),38.6 (C-21),136.9 (C-22),129.3 (C-23),129.5 (C-24),127.3 (C-25),129.5 (C-26),129.3 (C-27)。以上数据与文献[13]基本一致,故鉴定为金色酰胺醇酯。

化合物6:白色粉末,ESI-MS m/z:457.3[M-H]-。1H-NMR (CD3COCD3,600 MHz) δ:1.91(3H,s,H-29),0.88 (3H,s,H-23),0.83 (3H,s,H-27),0.81 (3H,s,H-26),0.72 (3H,s,H-25),0.62 (3H,s,H-24)；13C-NMR (CD3COCD3,150 MHz) δ:38.6 (C-1),27.4 (C-2),77.6 (C-3),38.7 (C-4),55.4 (C-5),18.2 (C-6),34.2(C-7),40.5 (C-8),50.5 (C-9),37.1 (C-10),20.8 (C-11),27.0 (C-12),37.2 (C-13),42.1(C-14),29.5 (C-15),31.4 (C-16),55.7 (C-17),48.8 (C-18),51.1 (C-19),210.3 (C-20),28.3 (C-21),36.4 (C-22),27.6 (C-23),15.1(C-24),15.7 (C-25),15.5 (C-26),14.1 (C-27),176.5 (C-28),29.6 (C-29)。以上数据与文献[14]基本一致,故鉴定化合物为platanic acid。

化合物7:橙红色针状结晶,ESI-MS m/z:229.1 [M+H]+。1H-NMR (CD3OD,600 MHz) δ:7.33 (2H,d,J=8.2 Hz,H-2′,6′),6.94(1H,d,J=16.2 Hz,β-H),6.78 (1H,d,J=16.2 Hz,α-H),6.75 (2H,d,J=8.3 Hz,H-3′,5′),6.44 (2H,d,J=2.0 Hz,H-2,6),6.15 (1H,s,H-4)；13C-NMR (CD3OD,150 MHz)δ:158.1 (C-3,5),156.9 (C-4′),139.9 (C-1),128.9 (C-1′),128.0 (C-β),127.2 (C-2′,6′),125.4 (C-α),114.5 (C-3′,5′),104.1(C-2,6),101.1 (C-4)。以上数据与文献[15]基本一致,故鉴定为反式白藜芦醇。

化合物8:白色粉末,ESI-MS m/z:525.3[M+H]+。1H-NMR (CDCl3,600 MHz) δ:4.87(1H,t,J=2.4 Hz,H-1),1.84～1.87 (1H,m,H-2a),1.70～1.72 (1H,m,H-2b),1.73～1.75(1H,m,H-3a),1.04～1.07 (1H,m,H-3b),5.38 (1H,d,J=9.0 Hz,H-6),5.58 (1H,t,J=9.0 Hz,H-7),2.23～2.25 (1H,m,H-8),2.60 (1H,t,J=12.0 Hz,H-9),1.65～1.67(1H,m,H-11a),2.02～2.04 (1H,m,H-11b),5.85 (1H,s,H-15),1.54 (1H,s,H-17),1.05(1H,s,H-18),1.06 (1H,s,H-19),1.10 (1H,s,H-20),1.99 (1-OAc,s),1.90 (6-OCOOH,s),2.08 (7-OCOOH,s)；13C-NMR (CDCl3,150 MHz) δ:71.5 (C-1),21.5 (C-2),31.4 (C-3),37.5 (C-4),78.3 (C-5),74.0 (C-6),74.4(C-7),49.5 (C-8),33.0 (C-9),43.6 (C-10),36.7 (C-11),103.6 (C-12),174.4 (C-13),73.9 (C-14),113.0 (C-15),169.0 (C-16),19.7 (C-17),29.5 (C-18),23.5 (C-19),15.8(C-20),20.3,168.7 (1-OCOOH),20.5,169.4(6-OCOOH),20.3,169.8 (7-OCOOH)。以上数据与文献 [16 ]基本一致,故鉴定为neocaesalpin L。

化合物9:白色粉末。1H-NMR (CD3OD,600 MHz) δ:5.37 (1H,brs,H-6),5.08 (1H,d,J=7.7 Hz,H-1′)；13C-NMR (CD3OD,150 MHz)δ:37.5 (C-1),30.2 (C-2),78.7 (C-3),40.0(C-4),140.9 (C-5),123.8 (C-6),34.2 (C-7),32.1 (C-8),50.3 (C-9),37.0 (C-10),21.3(C-11),40.3 (C-12),42.5 (C-13),56.8 (C-14),24.5 (C-15),28.6 (C-16),57.0 (C-17),12.2 (C-18),19.4 (C-19),36.4 (C-20),19.0(C-21),138.9 (C-22),129.5 (C-23),46.1 (C-24),29.5 (C-25),20.0 (C-26),19.2 (C-27),23.4 (C-28),12.0 (C-29),103.0 (C-1′),75.3(C-2′),78.5 (C-3′),71.7 (C-4′),78.1 (C-5′),62.9 (C-6′)。以上数据与文献[17]基本一致,故鉴定为Δ～ (5,22) 豆甾醇-3-O-β-D-吡喃葡萄糖苷。

化合物10:白色固体,TLC 喷洒10% 硫酸乙醇显紫红色。与豆甾醇标准对照品共薄层,在3 种不同的展开系统中比较Rf值完全一致,且显色也相同,故鉴定为豆甾醇。

化合物11:白色粉末。1H-NMR (CD3OD,600 MHz) δ:5.36 (H,m,H-6),5.07 (H,d,J=7.5 Hz,H-1′),4.59～3.94 (m,H-2′～6′),2.73～1.01 (m),0.98～0.82 (m,5 ×-CH3),0.67 (s,-CH3)；13C-NMR (CD3OD,150 MHz)δ:37.5 (C-1),30.3 (C-2),78.5 (C-3),39.4 (C-4),140.9 (C-5),122.0 (C-6),32.2 (C-7),32.1(C-8),50.4 (C-9),37.0 (C-10),21.3 (C-11),40.0 (C-12),42.5 (C-13),56.5 (C-14),24.5(C-15),28.6 (C-16),56.3 (C-17),12.0 (C-18),19.2 (C-19),36.4 (C-20),19.0 (C-21),30.8 (C-22),26.2 (C-23),46.1 (C-24),29.5(C-25),19.5 (C-26),20.0 (C-27),23.4 (C-28),12.2 (C-29),102.6 (C-1′),75.4 (C-2′),78.7 (C-3′),71.7 (C-4′),78.1 (C-5′),62.9 (C-6′)。以上数据与文献[18]基本一致,故鉴定为胡萝卜苷。

4 讨论

前期云实属植物化学成分研究的文献表明,一般是种子富含二萜成分,三萜成分较少；而茎枝正好相反,三萜成分较多,二萜成分较少。本研究分离鉴定的木栓酮、白桦脂酸、platanic acid 为三萜成分,仅neocaesalpin L 为二萜成分,与该属前期文献报道基本一致。从南蛇风茎中分离得到的白桦脂酸[19]、金色酰胺醇酯[13]、反式白藜芦醇[20]等成分具有抗炎、抗病毒活性,与其功效相一致,因此对其进行化学成分研究,以期为今后开发该植物的药用价值以及临床疗效提供理论基础。