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紫花前胡中化学成分的研究

2017-08-30廖志超姜鑫田文静林挺陈海峰

中国中药杂志 2017年15期
关键词:香豆素化学成分

廖志超+姜鑫+田文静+林挺 陈海峰

[摘要]采用HP20大孔吸附树脂、ODS、硅胶柱色谱、Sephadex LH20柱色谱和半制备HPLC等色谱分离手段对紫花前胡的化学成分进行研究。结合理化性质及MS, NMR等谱学数据鉴定化合物的结构,从紫花前胡80%乙醇提取物中分离并鉴定了12个化合物,分别是(9R,10R)9acetoxy8,8dimethyl9,10dihydro2H,8Hbenzo[1,2b:3,4b′]dipyran2one10yl ester(1)、 补骨脂呋喃香豆精(2)、顺式3′,4′二千里光酰基3′,4′二氢邪蒿内酯(3)、(3′R,4′R)3′angeloyloxy4′senecioyloxy3′,4′dihydroseselicalipteryxin(4)、(+)8,9dihydro8(2hydroxypropan2yl)2oxo2Hfuro[2,3h]chromen9yl3methylbut2enoate(5)、libanoridin(6)、丝立尼亭(7)、花椒素(8)、crocatone(9)、peujaponisinol B(10)、 peujaponisinol A(11)、ostenol(12)。其中,化合物1~5为首次从当归属植物中分离得到,化合物 7~12为首次从紫花前胡中分离得到。

[关键词]当归属; 紫花前胡; 香豆素; 化学成分

Chemical constituents from root of Angelica decursiva

LIAO Zhichao, JIANG Xin, TIAN Wenjing, LIN Ting*, CHEN Haifeng*

(School of Pharmaceutical Sciences, Xiamen University, Xiamen 361102, China)

[Abstract]The compounds were isolated and purified by HP20 macroporous adsorption resin, ODS, silica gel, and Sephadex LH20 column chromatography, as well as semipreparative HPLC chromatography from the 80% ethanol extract of the root of Angelica decursiva, and their structures were identified based on their physiochemical properties and spectroscopic data Twelve compounds were structures were identified as (9R,10R)9acetoxy8,8dimethyl9,10dihydro2H,8Hbenzo[1,2b:3,4b′]dipyran2one10yl ester (1), bakuchicin (2), (3′, S,4′S)disenecioyloxy3′,4′dihydroseselin (3), (3′R,4′R)3′angeloyloxy4′senecioyloxy3′,4′dihydroseselincalipteryxin (4), (+)8,9dihydro8(2hydroxypropan2yl)2oxo2Hfuro[2,3h]chromen9yl3methylbut2enoate (5), libanoridin (6), selinidin (7), suberosin (8), crocatone (9), peujaponisinol B (10), peujaponisinol A (11), and ostenol (12), respectively Compounds 15 were isolated from the plants of Angelica genus for the first time Compounds 712 were isolated from A decursiva for the first time

[Key words]Angelica; Angelica decursiva; coumarins; chemical constituents

中藥紫花前胡是伞形科Umbelliferae当归属Angelica植物紫花前胡A decursiva的干燥根。紫花前胡为多年生草本植物,根圆锥状,常数支根,表面黄褐色至棕色;具有疏散风热,降气化痰的功效[1]。紫花前胡中的主要化学成分为香豆素类化合物。香豆素是广泛分布于植物界中的次生代谢产物,最早报道于1820年,迄今已有1 300多种[2]。现代药理学研究表明,香豆素类化合物具有抗神经衰弱、抗凝血、抗氧化、抗菌、抗癌、抗结核、降血糖、抗抑郁、抗炎等多方面的药理活性[3],在药物的研究开发中发挥了重要的作用。本文以富含香豆素类成分的紫花前胡为研究对象,从其80%乙醇提取物中分离鉴定了12个化合物。

1材料

Thermo Scientific Q Exactive质谱仪(赛默飞公司);Bruker Avance 600Ⅲ 型核磁共振仪(瑞士Bruker公司);LC20A 分析型高效液相色谱仪(日本岛津公司);LC8A制备型高效液相色谱仪(日本岛津公司);半制备HPLC色谱柱(北京绿百草科技发展有限公司 Prep C18,10 mm×150 mm,5 μm) ; Sephadex LH20 填料(Pharmacia) ; ODS(40~63 μm,Pharmacia); 大孔树脂型号为 HP20(北京绿百草科技发展有限公司) ; 柱色谱用硅胶(200~300目) 及薄层色谱用GF254硅胶预制板(青岛海洋化工厂)。有机溶剂均为分析纯或色谱纯。

紫花前胡饮片经永春县林业局邹秀红工程师鉴定为伞形科当归属植物紫花前胡A decursiva的干燥根。

2 提取分离

紫花前胡根饮片100 kg,以80%乙醇回流提取2次,每次3 h,合并乙醇提取液,浓缩得到浸膏20 kg。用蒸馏水混悬浸膏后,利用大孔树脂进行不同极性部分的分离。用乙醇水进行洗脱,得到水洗脱部分、20%乙醇部分、60%乙醇部分和95%乙醇部分。将95%乙醇洗脱部分(4141 g)进行硅胶柱色谱分离,用氯仿甲醇系统(100∶0~0∶1)进行梯度洗脱,得到10个流分(Fr D1~D10)。Fr D1用水混悬,依次用石油醚和氯仿萃取,得到氯仿萃取物Fr D1B(903 g)。Fr D1B经硅胶柱色谱,用石油醚丙酮(30∶1~20∶1)进行梯度洗脱,得到7个流分(Fr D1B1~D1B7)。Fr D1B3 经Sephadex LH20柱色谱、反相柱色谱以及半制备HPLC纯化得到化合物2(85 mg) 和其他6个流分(Fr D1B3A1~D1B3A7)。Fr D1B3A3经半制备HPLC纯化得到化合物9(29 mg);Fr D1B3A5经半制备HPLC纯化得到化合物7(79 mg)和 8(22 mg);Fr D1B6经反相柱色谱以及半制备HPLC纯化得到化合物10(20 mg),11(28 mg),12(45 mg);Fr D1B5经反相柱色谱,依次用甲醇水(60∶40~80∶20)梯度洗脱,得到10个流分(Fr D1B5A~D1B5J);Fr D1B5D经半制备HPLC纯化得到化合物6 (58 mg)和 1(71 mg);Fr D1B5E经半制备HPLC纯化得到化合物5(60 mg);Fr D1B5H经反相柱色谱得到5个流分(Fr D1B5H1~D1B5H5);Fr D1B5H2经半制备HPLC纯化得到化合物3(428 mg);Fr D1B5H5经半制备HPLC纯化得到化合物4(185 mg)。

3结构鉴定

化合物1无色油状,ESIMS m/z 383 [M+Na]+。1HNMR(CDCl3,600 MHz) δ: 624(1H,d,J=95 Hz,H3),763(1H,d,J=95 Hz,H4),735(1H,d,J=83 Hz,H5),681(1H,d,J=83 Hz,H6),530(1H,d,J=47 Hz,H3′),655(1H,d,J=47 Hz,H4′),209(3H,s,H2″),245(1H,dq,J=160,76 Hz H2),242(1H,dq,J=160,76 Hz H2),120(3H,dd,J=76 Hz,H3),145(3H,s,H5′),141(3H,s,H6′)。13CNMR(CDCl3,150 MHz) δ: 1599(C2),1132(C3),1433(C4),1126(C10),1292(C5),1144(C6),1566(C7),1070(C8),1540(C9),775(C2′),702(C3′),608(C4′),1700(C1″),207(C2″),1734(C1),275(C2),91(C3),227(C5′),248(C6′)。以上數据与文献[4]对比,确定该化合物为(9R,10R)9acetoxy8,8dimethyl9,10dihydro2H,8Hbenzo[1,2b:3,4b′]dipyran2one10yl ester。

化合物2无色针状,ESIMS m/z 209 [M+Na]+。1HNMR(CDCl3,600 MHz) δ: 640(1H,d,J=95 Hz,H3),781(1H,d,J=95 Hz,H4),744(1H,d,J=83 Hz,H7),738(1H,d,J=83 Hz,H8),770(1H,d,J=19 Hz,H2′),714(1H,d,J=19 Hz,H3′)。13CNMR(CDCl3,150 MHz) δ: 1609(C2),1445(C3),1141(C4),1169(C10),1574(C5),1135(C6),1238(C7),1088(C8),1485(C9),1041(C2′),1459(C3′)。以上数据与文献[5]对比,确定该化合物为补骨脂呋喃香豆精。

化合物3白色粉末,ESIMS m/z 449 [M+Na]+。1HNMR(CDCl3,600 MHz) δ: 620(1H,d,J=95 Hz,H3),759(1H,d,J=95 Hz,H4),734(1H,d,J=84 Hz,H5),680(1H,d,J=84 Hz,H6),540(1H,d,J=49 Hz,H3′),662(1H,d,J=49 Hz,H4′),144(3H,s,H5′),142(3H,s,H6′),567(1H,q,J=12 Hz,H2″),188(3H,d,J=09 Hz,H4″),187(3H,d,J=09 Hz,H5″),562(1H,q,J=12 Hz,H2),219(3H,d,J=08 Hz,H4),215(3H,d,J=08 Hz,H5)。13CNMR(CDCl3,150 MHz) δ: 1599(C2),1131(C3),1432(C4),1125(C10),1290(C5),1144(C6),1568(C7),1076(C8),1540(C9),777(C2′),694(C3′),598(C4′),251(C5′),226(C6′),1651(C1″),1152(C2″),1575(C3″),274(C4″),203(C5″),1652(C1),1153(C2),1582(C3),274(C4),203(C5)。以上数据与文献[6]对比,确定该化合物为顺式3′,4′二千里光酰基3′,4′二氢邪蒿内酯。

化合物4白色粉末,ESIMS m/z 449 [M+Na]+。1HNMR(CDCl3,600 MHz) δ: 621(1H,d,J=95 Hz,H3),758(1H,d,J=95 Hz,H4),735(1H,d,J=86 Hz,H5),680(1H,d,J=86 Hz,H6),540(1H,d,J=49 Hz,H3′),665(1H,d,J=49 Hz,H4′),144(3H,s,H5′),149(3H,s,H6′),611(1H,qq,J=74,13 Hz,H2″),196(3H,dq,J=74,13 Hz,H4″),185(3H,q,J=12 Hz,H5″),561(1H,m,H2),219(3H,d,J=12 Hz,H4),187(3H,d,J=13 Hz,H5)。13CNMR(CDCl3,150 MHz) δ: 1599(C2),1133(C3),1432(C4),1125(C10),1291(C5),1144(C6),1567(C7),1076(C8),1540(C9),775(C2′),703(C3′),595(C4′),255(C5′),225(C6′),1663(C1″),1272(C2″),1394(C3″),158(C4″),204(C5″),1659(C1),1151(C2),1579(C3),274(C4),204(C5)。以上数据与文献[7] 对比,确定该化合物为(3′S,4′S)disenecioyloxy3′,4′dihydroseselin。

化合物5无色油状,ESIMS m/z 367 [M+Na]+。1HNMR(CDCl3,600 MHz) δ: 621(1H,d,J=95 Hz,H3),760(1H,d,J=95 Hz,H4),740(1H,d,J=84 Hz,H5),690(1H,d,J=84 Hz,H6),450(1H,d,J=65 Hz,H2′),700(1H,d,J=65 Hz,H3′),142(3H,s,H2″),140(3H,s,H3″),560(1H,brs,H2),190(3H,s,H4),220(3H,s,H5)。13CNMR(CDCl3,150 MHz) δ: 1599(C2),1131(C3),1436(C4),1133(C10),1314(C5),1078(C6),1639(C7),1129(C8),1517(C9),917(C2′),683(C3′),711(C1″),269(C2″),261(C3″),1648(C1),1145(C2),1615(C3),277(C4),207(C5)。以上数据与文献[8] 对比,確定该化合物为(+)8,9dihydro8(2hydroxypropan2yl)2oxo2Hfuro[2,3 h]chromen9yl3methylbut2enoate。

化合物6白色粉末,ESIMS m/z 289 [M+H]+。1HNMR(CDCl3,600 MHz) δ: 621(1H,d,J=95 Hz,H3),763(1H,d,J=95 Hz,H4),726(1H,d,J=83 Hz,H5),675(1H,d,J=83 Hz,H6),516(1H,dd,J=100,80 Hz,H2′),338(1H,dd,J=160,100 Hz,H3′),328(1H,dd,J=160,80 Hz,H3′),199(3H,s,H4″),157(3H,s,H1″CH3),151(3H,s,H1″CH3)。13CNMR(CDCl3, 150 MHz) δ: 1610(C2),1123(C3),1439(C4),1131(C10),1289(C5),1067(C6),1639(C7),1134(C8),1513(C9),887(C2′),276(C3′),821(C1″),1703(C3″),219(C4″),223(C1″CH3),209(C1″CH3)。以上数据与文献[9] 对比,确定该化合物为libanoridin。

化合物7白色粉末,ESIMS m/z 351 [M+Na]+。1HNMR(CDCl3,600 MHz) δ: 623(1H,d,J=95 Hz,H3),763(1H,d,J=95 Hz,H4),724(1H,d,J=82 Hz,H5),679(1H,d,J=82 Hz,H6),521(1H,t,J=53 Hz,H3′),325(1H,dd,J=178,52 Hz,H4′),310(1H,dd,J=178,53 Hz,H4′),610(1H,qq,J=73,14 Hz,H3″),191(3H,dq,J=73,14 Hz,H4″),185(3H,q,J=14 Hz,H5),139(3H,s,H5′),138(3H,s,H6′)。13CNMR(CDCl3,150 MHz) δ: 1612(C2),1125(C3),1438(C4),1121(C10),1267(C5),1142(C6),1564(C7),1073(C8),1534(C9),766(C2′),693(C3′),232(C4′),248(C5′),228(C6′),1668(C1″),1274(C2″),1392(C3″),158(C4″),206(C5″)。以上数据与文献[10] 对比,确定该化合物为丝立尼亭。

化合物8无色针状,ESIMS m/z 245 [M+H]+。1HNMR(CDCl3,600 MHz) δ: 623(1H,d,J=95 Hz,H3),762(1H,d,J=95 Hz,H4),718(1H,s,H5),678(1H,s,H8),331(2H,d,J=74 Hz,H2′),528(1H,tq,J=73,13 Hz,H3′),177(3H,s,4′CH3),170(3H,s,4′CH3),390(3H,s,7OCH3)。13CNMR(CDCl3,150 MHz) δ: 1616(C2),1128(C3),1439(C4),1119(C10),1274(C5),1275(C6),1607(C7),985(C8),1545(C9),278(C2′),1213(C3′),1337(C4′),258(C4′),178(C5′),559(7OCH3)。以上数据与文献[11] 对比,确定该化合物为花椒素。

化合物9无色针状。ESIMS m/z 209 [M+H]+。1HNMR(CDCl3,600 MHz) δ: 714(1H,d,J=12 Hz,H2),727(1H,d,J=12 Hz,H4),606(2H,s,H7),292(2H,q,J=72 Hz,H2′),121(3H,t,J=72 Hz,H3′),395(3H,s,H1OCH3)。13CNMR(CDCl3,150 MHz) δ: 1393(C1),1026(C2),1319(C3),1086(C4),1435(C5),1023(C7),1489(C9),1989(C1′),316(C2′),849(C3′),566(C1OCH3)。以上数据与文献[12] 对比,确定该化合物为crocatone。

化合物10无色油状,ESIMS m/z 367 [M+Na]+。1HNMR(CDCl3,600 MHz) δ: 622(1H,d,J=95 Hz,H3),760(1H,d,J=95 Hz,H4),733(1H,d,J=86 Hz,H5),679(1H,d,J=86 Hz,H6),414(1H,d,J=47 Hz,H3′),646(1H,d,J=47 Hz,H4′),147(3H,s,H5′),144(3H,s,H6′),572(1H,m,H2″),224(3H,s,H5″),192(3H,s,H6″)。13CNMR(CDCl3,150 MHz) δ: 1606(C2),1125(C3),1439(C4),1125(C10),1287(C5),1146(C6),1561(C7),1107(C8),1542(C9),778(C2′),714(C3′),602(C4′),255(C5′),226(C6′),1656(C1″),1152(C2″),1592(C3″),276(C5″),205(C6″)。以上数据与文献[13] 对比,确定该化合物为peujaponisinol B。

化合物11无色针状,ESIMS m/z 367 [M+Na]+。1HNMR(600 MHz,CDCl3) δ: 624(1H,d,J=95 Hz,H3),763(1H,d,J=95 Hz,H4),733(1H,d,J=86 Hz,H5),679(1H,d,J=86 Hz,H6),521(1H,d,J=47 Hz,H3′),543(1H,d,J=47 Hz,H4′),149(3H,s,H5′),142(3H,s,H6′),580(1H,m,H2″),220(3H,s,H5″),192(3H,s,H6″)。13CNMR(CDCl3,150 MHz) δ: 1601(C2),1130(C3),1433(C4),1123(C10),1292(C5),1145(C6),1570(C7),1072(C8),1542(C9),787(C2′),717(C3′),629(C4′),255(C5′),212(C6′),1675(C1″),1149(C2″),1598(C3″),277(C5″),206(C6″)。以上数据与文献[14] 对比,确定该化合物为peujaponisinol A。

化合物12白色粉末,ESIMS m/z 231 [M+H]+。1HNMR(CDCl3,600 MHz) δ: 625(1H,d,J=95 Hz,H3),763(1H,d,J=95 Hz,H4),723(1H,d,J=86 Hz,H5),680(1H,d,J=86 Hz,H6),362(2H,d,J=72 Hz,H2′),528(1H,m,H3′),176(3H,br s,H5′),183(3H,br s,H6′)。13CNMR(CDCl3,150 MHz) δ: 1616(C2),1132(C3),1441(C4),1127(C10),1266(C5),1125(C6),1362(C7),1147(C8),1583(C9),221(C2′),1203(C3′),1531(C4′),258(C5′),180(C6′)。以上数据与文献[15] 对比,确定该化合物为ostenol。

4小结

本研究从紫花前胡中分离得到12 个化合物,其中化合物1~5为首次从当归属植物中分离得到,化合物 7~12为首次从紫花前胡中分离得到,丰富了紫花前胡香豆素类成分的结构多样性。前人研究发现化合物1可以发挥抗多重耐药作用,且与多重耐药抑制剂verapamil 和cyclosporine A活性相当[16];化合物2具有血管舒张活性[16],抗氧化活性[17]和对HepG2肿瘤细胞抑制作用[18];化合物3具有抗炎[19],抑制黑色素瘤的作用[20];化合物6可以调节炎症过敏反应[21],治疗由于紫外线照射造成的皮肤损伤[22],另外还可以发挥抗炎作用[23];化合物7可以调节平滑肌收缩从而调节血管和肌肉的舒张[24],抗真菌[25]和抗炎作用[26];化合物8对杜什曼虫前鞭毛体有作用[27],还可以发挥抗炎活性[28]。因此,本研究可以为这些化合物的制备提供新的植物来源。

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