严 岚,金慧子＊,聂利月,覃江江,付建军,张卫东,2

1上海交通大学药学院,上海 200240;2上海第二军医大学药学院,上海 200433

显脉旋覆花化学成分的研究

严 岚1,金慧子1＊,聂利月1,覃江江1,付建军1,张卫东1,2

1上海交通大学药学院,上海 200240;2上海第二军医大学药学院,上海 200433

从显脉旋覆花 (Inula nervosaWall.)地上部分的 75%乙醇提取物中分离到 13个化合物,经波谱鉴定为紫菀酮 (1),β-谷甾醇 (2),α-菠菜甾醇 (3),熊果酸 (4),胡萝卜苷 (5),bigelovin(6),loliolide(7),24S-乙基 -5α-胆甾 -7,22E-二烯 -3α-醇 -β-半乳糖苷 (8),菠叶素 (9),山萘酚 (10),α-菠甾醇 -3-O-β-D-葡萄糖苷 (11),苄醇 -β-D-葡萄糖苷 (12)和 2-苯乙醇-β-D-葡萄糖苷 (13)。其中,化合物 1～3和 6～13为首次从该植物中分离得到。

显脉旋覆花;菊科;化学成分;分离

Abstract:Thirteen compoundswere isolated from the 75%ethanol extract of the aerial parts ofInula nervosaWall..On the basis of spectral data,theywere identified as shionone(1),β-sitosterol(2),α-spinasterol(3),ursolic acid(4),daucosterol(5),bigelovin(6),loliolide(7),24S-ethyl-5α-cholesta-7,22E-dien-3α-ol-β-galactopyranoside(8),spinacetin(9),kaempferol(10),α-spinasterol-3-O-β-D-glucoside(11),benzyl-β-D-glucopyranoside(12),and 2-phenylethyl-β-D-glucopyranoside(13).Compounds 1-3 and 6-13 were isolated from this plant for the first time.

Key words:Inula nervosaWall.;Asteraceae;chemical constituents;isolate

Introduction

Inula nervosaWall.(Asteraceae)is a perennial tree,mainly distributed in southwest China.The roots of this plant have been used asChinese folk medicine for treating stomachache and relieving rheumatis m[1].However,up to now,only a few chemical constituents,such as daucosterol[2],oleanolic acid[3],stigmasterol[2,3],thymol[2],have been isolated from this plant.In this study on the aerial parts ofI.nervosa,thirteen compounds were isolated and identified as shionone(1),β-sitosterol(2),α-spinasterol(3),ursolic acid(4),daucosterol(5),bigelovin(6),loliolide(7),24S-ethyl-5α-cholesta-7, 22E-dien-3α-ol-β-galactopyranoside(8),spinacetin(9),kaempferol(10),α-spinasterol-3-O-β-D-glucoside(11),benzyl-β-D-glucopyranoside(12),and 2-phenylethyl-β-D-glucopyranoside(13).Compounds 1-3 and 6-13 were isolated from this plant for the first time.

Exper imental

General procedures

All melting points were determined on an RY-1 micromelting point apparatus(uncorrected).The ES IMS were measured on an Agilent 1100 series mass spectrometer,whereas E IMS were measured on an Autospec-Ult imaE TOF apparatus.NMR spectra were measured on a Bruker DRX-500 spectrometer,operating at 500 MHz for1H and 125 MHz for13C NMR.Chemical shift(δ)were given in ppm relative to T MS as internal standard and coupling constants in Hz.Silica gel(200-300 mesh,Yantai,China)and sephadex LH-20(GE Healthcare Bio-Sciences AB,Sweden)were used for column chromatography,and precoated silica GF254plateswere used for TLC(Qingdao Haiyang Chemical Co.,Ltd.,China).

Plantmaterial

The aerial parts ofI.nervosawere collected from Yunnan Province of China in July,2006 and identified by Prof.Huang Bao-kang,Department of Phar macognosy,School of Phar macy,Second MilitaryMedical University.A voucher specimen was deposited at School of Pharmacy,Shanghai Jiao TongUniversity.

Extraction and isolation

The air-dried and powdered aerial parts ofI.nervosa(15.0 kg)were extracted with 75%EtOH for three times at room temperature.The solvents were concentrated under vacuum to yield a residue(1361.0 g).The residue was dissolved in water(5.0 L)and extracted successively with petroleum ether(5.0 L×10),ethyl acetate(5.0 L×10)and n-butanol(5.0 L×10)to give corresponding fractionsA(200.2 g),B(277.6 g),and C(291.7 g).Fraction A was subjected to silica gel column chromatographywith petroleum ether:ethyl acetate(100∶1-0∶100)to give 16 fractions A1-A16.FractionsA5,A8,A9,A12 and A16 were purified by recrystallization from CH2Cl2to yield compounds 1(63.3 mg),2(50.5 mg),3(120.0 mg),4(8.6 mg),and 5(141.5 mg),respectively.Fraction B was subjected to silica gel column chromatographywith CH2Cl2∶MeOH(100∶1-0∶100)to give 9 fractionsB1-B9.Fraction B3 was separated on silica gel column eluted by petroleum ether-ethyl acetate(10∶1)to afford 6(8.2 mg)and 7(9.4 mg).Fraction B5 was purified by recrystallization from Me2CO to yield 8(5.0 mg).Compounds 9(11.9 mg)and 10(3.0 mg)were obtained from the residue of B5 by column chromatography on sephadex LH-20 eluted with MeOH.Eleven subfractionsB6a-B6k and compound 11(551.0 mg)were obtained from fraction B6 separated over silica gel column eluted with CH2Cl2∶MeOH(10∶1-1∶1).Subfraction B6c was separated over silica gel column using CH2Cl2-MeOH(10∶1)to give compounds 12(18.6 mg)and 13(7.0 mg).

Structure Identification

Shionone(1) C30H50O,whiteneedlecrystal(CH2Cl2),mp.159-160℃;EI-MSm/z(%):426[M]+(80),411[M-CH3]+(20),341(50),109(70),95(70),69(100);1H NMR(CDCl3)δ:5.10(1H,m,H-21),1.69(3H,s,H-29),1.61(3H,s,H-30),1.14(3H,s,H-27),0.93(3H,s,H-25),0.91(3H,s,H-28),0.90(3H,s,H-26),0.88(3H,d,J=6.6 Hz,H-23),0.72(3H,s,H-24);13C NMR(CDCl3)δ:213.3(C-3),130.8(C-22),125.2(C-21),59.6(C-10),58.2(C-4),49.9(C-8),44.5(C-18),43.6(C-19),42.2(C-5),41.4(C-2),41.1(C-6),38.6(C-14),38.5(C-9),36.9(C-13),35.3(C-11),34.6(C-16),32.9(C-28),32.3(C-12),31.7(C-17),29.2(C-15),25.7(C-29),23.2(C-20),22.3(C-1),20.6(C-27),19.6(C-25),17.9(C-7),17.6(C-30),15.2(C-26),14.6(C-24),6.8(C-23).The NMR andMS data were in accordance with those reported in the literature[4],and identified 1 as shionone.

β-Sitosterol(2) C29H50O,white needle crystal(CH2Cl2),mp.141-143℃.TLC behavior was identicalwith that of authenticβ-sitosterol.

α-Spinasterol(3) C29H48O,white needle crystal(CH2Cl2),mp.160-162℃;ESI-MSm/z:413[M+H]+;1H NMR(CDCl3)δ:5.18(1H,m,H-22),5.15(1H,m,H-7),5.03(1H,m,H-23),3.60(1H,m,H-3),1.03(3H,d,J=7.0 Hz,H-21),0.86(3H,d,J=7.0 Hz,H-26),0.82(3H,m,H-29),0.81(3H,s,H-19),0.80(3H,d,J=7.0 Hz,H-27),0.57(3H,s,H-18);13C NMR(CDCl3)δ:139.6(C-8),138.2(C-22),129.5(C-23),117.5(C-7),71.1(C-3),55.9(C-17),55.1(C-14),51.3(C-24),49.5(C-9),43.3(C-13),40.8(C-20),40.3(C-5),39.5(C-12),38.0(C-4),37.2(C-1),34.2(C-10),31.9(C-25),31.5(C-2),29.6(C-6),28.5(C-16),25.4(C-28),23.0(C-15),21.6(C-11),21.4(C-21),21.1(C-26),19.0(C-27),13.0(C-19),12.2(C-18),12.0(C-29).The NMR and MS data were in accordance with those reported in the literature[5],and identified 3 as α-spinasterol.

Ursolic acid(4) C30H48O3,white amorphous powder,mp.258-259℃;TLC behavior was identical with that of authentic ursolic acid.

Daucosterol(5) C35H60O6,white amorphous powder,mp.285-287℃;TLC behavior was identical with that of authentic daucosterol.

Bigelovin(6) C17H20O5,white needle crystal(CH2Cl2),mp.180-182℃;ESI-MSm/z:327[M+Na]+;1H NMR(CDCl3)δ:7.10(1H,dd,J=6.0,1.5 Hz,H-2),6.22(1H,d,J=3.0 Hz,H-13a),6.11(1H,dd,J=6.0,3.0 Hz,H-3),5.92(1H,d,J=3.0 Hz,H-13b),5.61(1H,d,J=7.5 Hz,H-6),4.61(1H,dt,J=12.0,3.0 Hz,H-8),3.07(1H,m,H-7),3.03(1H,m,H-1),2.57(1H,m,H-9a),2.06(1H,m,H-10),1.97(3H,s,H-17),1.54(1H,m,H-9b),1.29(3H,d,J=6.5 Hz,H-14),1.21(3H,s,H-15);13C NMR(CDCl3)δ:209.1(C-4),169.6(C-16),168.9(C-12),162.5(C-2),137.1(C-11),130.8(C-3),122.1(C-13),76.1(C-8),73.1(C-6),56.2(C-5),53.9(C-7),52.2(C-1),44.4(C-9),27.2(C-10),22.6(C-15),21.0(C-17),19.7(C-14).The above data were identical with the literature’s[6].

Loliolide(7) C11H16O3,whiteneedlecrystal(CH2Cl2),mp.156-158℃;ESI-MSm/z:219[M+Na]+;1H NMR(CDCl3)δ:5.69(1H,s,H-7),4.32(1H,t,J=3.5 Hz,H-3),2.46(1H,dt,J=14.0,2.5 Hz,H-4a),1.97(1H,dt,J=14.0,2.5 Hz,H-2a),1.77(1H,m,H-4b),1.78(3H,s,H-11),1.52(1H,dd,J=14.5,3.5 Hz,H-2b),1.47(3H,s,H-9),1.29(3H,s,H-10);13C NMR(CDCl3)δ:182.6(C-6),172.0(C-8),112.8(C-7),86.8(C-5),66.7(C-3),47.3(C-2),45.6(C-4),35.9(C-1),30.6(C-9),27.0(C-11),26.4(C-10).The above data were identicalwith the literature’s[7].

24S-Ethyl-5α-cholesta-7,22E-dien-3α-ol-β-galactopyranoside(8) C35H58O6,white powder,mp.290-291℃;ESI-MSm/z:575[M+H]+;1H NMR(C5D5N)δ:5.17(1H,m,H-7),5.14(1H,m,H-22),5.03(1H,dd,J=15.0,4.0 Hz,H-23),3.57(1H,m,H-3),1.03(3H,d,J=6.5 Hz,H-21),0.86(3H,d,J=6.0 Hz,H-26),0.85(3H,m,H-29),0.79(3H,s,H-19),0.78(3H,d,J=6.5 Hz,H-27),0.55(3H,s,H-18);Galactose:4.49(1H,dd,J=12.0,4.5 Hz,H-6′a),4.39(1H,d,J=8.0 Hz,H-1′),4.27(1H,dd,J=12.0,2.5 Hz,H-6′b),3.60(2H,m,H-3′,H-4′),3.45(1H,m,H-5′),3.36(1H,m,H-2′);13C NMR(C5D5N)δ:139.6(C-8),138.1(C-22),129.5(C-23),117.3(C-7),78.9(C-3),55.9(C-17),55.1(C-14),51.3(C-24),49.4(C-9),43.2(C-13),40.8(C-20),40.2(C-5),39.4(C-12),37.1(C-1),34.4(C-10),34.2(C-4),31.9(C-25),29.5(C-2),29.2(C-6),28.5(C-16),25.4(C-28),23.0(C-15),21.5(C-11),21.4(C-21),21.1(C-26),19.0(C-27),13.0(C-19),12.2(C-29),12.0(C-18);Galactose:101.1(C-1′),76.0(C-3′),74.0(C-5′),73.6(C-2′),70.1(C-4′),62.7(C-6′).The NMR and MS data were in accordance with those reported in the literature[8],and identified 8 as 24S-ethyl-5α-cholesta-7,22E-dien-3αol-β-galactopyranoside.

Spinacetin(9) C17H14O8,yellow powder,mp.122-124℃;ESI-MSm/z:347[M +H]+;1H NMR(CD3OD)δ:7.61(1H,d,J=2.0 Hz,H-2′),7.51(1H,dd,J=8.0,2.0 Hz,H-6′),6.89(1H,d,J=8.0 Hz,H-5′),6.48(1H,s,H-8),3.88(3H,s,3′-OCH3),3.78(3H,s,6-OCH3);13C NMR(CD3OD)δ:180.3(C-4),158.7(C-7),158.1(C-9),153.8(C-5),153.7(C-2),150(C-4′),146.5(C-3′),139.2(C-3),132.6(C-6),123(C-1′),122.4(C-6′),116.5(C-5′),116.4(C-2′),106.3(C-10),94.9(C-8),61.0(6-OCH3),60.5(3′-OCH3).The NMR and MS data were in accordance with those reported in the literature[9],and identified 9 as spinacetin.

Kaempferol(10) C15H10O6,yellow powder,mp.276-278℃;ESI-MSm/z:285[M-H]-;1H NMR(CD3OD)δ:8.09(2H,d,J=8.5 Hz,H-2′,6′),6.91(2H,d,J=8.5 Hz,H-3′,5′),6.40(1H,d,J=2.0 Hz,H-8),6.19(1H,d,J=2.0 Hz,H-6);13C NMR(CD3OD)δ:177.4(C-4),165.6(C-7),162.5(C-9),160.6(C-4′),158.3(C-5),148.0(C-2),137.2(C-3),130.7(C-2′,6′),121.6(C-1′),116.3(C-3′,5′),104.6(C-10),99.3(C-6),94.5(C-8).The NMR and MS data were in accordance with those reported in the literature[10],and identified 10 as kaempferol.

α-Spinasterol-3-O-β-D-glucoside(11) C35H58O6,white powder,mp.272-280℃;ESI-MSm/z:575[M+H]+;1H NMR(C5D5N)δ:5.28(1H,m,H-7),5.20(1H,m,H-22),5.15(1H,dd,J=15.0,3.5 Hz,H-23),3.60(1H,m,H-3),1.25(3H,d,J=6.5 Hz,H-21),0.95(3H,d,J=6.0 Hz,H-26),0.92(3H,d,J=6.5 Hz,H-27),0.91(3H,m,H-29),0.78(3H,s,H-19),0.63(3H,s,H-18);Glucose:5.00(1H,d,J=8.0 Hz,H-1′),4.52(1H,dd,J=12.5,2.0 Hz,H-6′b),4.37(1H,dd,J=12.5,5.0 Hz,H-6′a),4.22(2H,m,H-3′,H-4′),3.98(2H,m,H-2′,H-5′);13C NMR(C5D5N)δ:139.6(C-8),138.2(C-22),129.5(C-23),117.5(C-7),77.0(C-3),55.9(C-17),55.0(C-14),51.1(C-24),49.4(C-9),43.2(C-13),40.6(C-20),40.0(C-5),39.4(C-12),37.1(C-1),34.5(C-4),34.3(C-10),31.8(C-25),29.7(C-2),29.6(C-6),28.4(C-16),25.2(C-28),23.0(C-15),21.4(C-11),21.2(C-21),20.8(C-26),18.9(C-27),12.6(C-19),12.1(C-29),11.9(C-18);Glucose:102.0(C-1′),78.3(C-5′),77.9(C-3′),75.0(C-2′),71.6(C-4′),62.7(C-6′).The NMR andMS data were in accordance with those reported in the literature[11],and identified 11 asα-spinasterol-3-O-β-D-glucoside.

Benzyl-β-D-glucopyranoside(12) C13H18O6,colorless gum;ESI-MSm/z:293[M+Na]+;1H NMR(CD3OD)δ:7.42(2H,d,J=7.5 Hz,H-2,6),7.33(2H,m,H-3,5),7.27(1H,d,J=7.5 Hz,H-4),4.93(1H,d,J=12.0 Hz,H-7a),4.67(1H,d,J=12.0 Hz,H-7b);13C NMR(CD3OD)δ:139.1(C-1),129.3(C-3,5),129.2(C-2,6),128.7(C-4),71.8(C-7);Glucose:103.3(C-1′),78.1(C-3′),78.0(C-5′),75.1(C-2′),71.7(C-4′),62.8(C-6′).The NMR and MS data were in accordance with those reported in the literature[12],and identified 12 as benzyl-β-D-glucopyranoside.

2-Phenylethyl-β-D-glucopyranoside(13) C14H20O6,colorless gum;ESI-MSm/z:307[M+Na]+;1H NMR(CD3OD)δ:7.22(5H,m,H-2,3,4,5,6),4.10(1H,m,H-8a),3.74(1H,m,H-8b),2.94(2H,m,H-7);13C NMR(CD3OD)δ:140.1(C-1),130.0(C-2,6),129.3(C-3,5),127.2(C-4),71.7(C-8),37.2(C-7);Glucose:104.4(C-1′),78.1(C-3′),78.0(C-5′),75.1(C-2′),71.6(C-4′),62.7(C-6′).The above data were identicalwith the literature’s[13].

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Chemical Constituents from Inulanervosa Wall.

YAN Lan1,JIN Hui-zi1＊,NIE Li-yue1,QIN Jiang-jiang1,FU Jian-jun1,ZHANG Wei-dong1,2

1School of Phar macy,Shanghai Jiao Tong University,Shanghai 200240,China;2School of Phar macy,Second Military University,Shanghai 200433,China

R284.2;Q946.91

A

1001-6880(2011)02-0258-04

Received August 25,2009;Accepted October 21,2009

Foundation Item:This research project was supported by the Scientific Foundation of Shanghai China(08DZ1971302).

＊Corresponding author Tel:86-21-34205989;E-mail:wdzhangy@hotmail.com