旋覆花的化学成分研究

覃江江，朱佳娴，朱 燕，严 岚，金慧子*，张卫东，2*

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

Introduction

Inula japonica Thunb.，which belongs to Asteraceae，is widespread in Europe，Africa，Asia，and Mediterranean［1］.As one of the most important members in this genus，I.japonica has been reported to possess diverse biological activities，such as anti-tumor［2-4］，anti-diabetic［5］，and hypolipidemic［5］activities.During our search for bioactive compositions from this plant，anthranilic acid derivatives［6］and sesquiterpenes［2］have been reported by us before.In this paper，we described the isolation and structure elucidation of coumarins and other constituents which were isolated from this genus for the first time.

Experimental

General

1H and13C NMR spectra were recorded on Bruker DRX-500 spectrometers at 500 and 125 MHz，respectively.ESIMS spectra were recorded on Varian MAT-212 mass spectrometer.A preparative column(Shimadzu PRC-ODS EV0233)was used for preparative HPLC (Shimadzu LC-6AD).All solvents used were of analytical grade(Shanghai Chemical Company，Ltd.).TLC analysis was run on HSGF254silica gel plates(10-40 μm，Yantai，China).Column chromatography was performed on silica gel(100-200，200-300 mesh，Yantai，China)，silica gel H(10-40 μm，Qingdao，China)and Sephadex LH-20(Pharmacia Co.Ltd.).

Plant material

The aerial parts of I.japonica were collected in Anhui province，PR China，in October，2006，and were authenticated by Professor Huang Baokang，Department of Pharmacognosy，School of Pharmacy，Second Military MedicalUniversity.A voucher specimen (No.2007XFH1)was deposited at School of Pharmacy，Shanghai Jiao Tong University.

Extraction and isolation

The dried aerial parts of I.japonica(20.0 kg)were powdered and extracted with 95% ethanol for three times at room temperature.The ethanolic extract was successively partitioned with petroleum ether(PE)，CH2Cl2，EtOAc，and n-BuOH，respectively.The CH2Cl2fraction(84.5 g)was chromatographed on a silica gel column eluting with a step gradient of CH2Cl2· MeOH(100∶0，50∶1，20∶1，10∶1，5∶1，2∶1，1∶1)to give subfractions.Then subfractions were isolated and purified in a combination of sicila gel，Sephadex LH-20 and preparative HPLC to afford compounds 3(6.0 mg)，4(10.8 mg)，5(6.6 mg)，6(3.0 mg)，7(3.0 mg)，8(16.0 mg)，9(4.0 mg)，and 10(26.0 mg).And from PE fraction(100.8 g)，1(4.0 mg)and 2 (6.5 mg)were obtained.

Results and Discussion

Ayapin(1)White amorphous powder，ESI-MS m/z 213［M+Na］+，189［M-H］-.1H NMR(CDCl3，500 MHz)δ:7.58(1H，d，J=10.0 Hz，H-4)，6.83(2H，s，H-5，8)，6.28(1H，d，J=10.0 Hz，H-3)，6.07 (2H，s，H-11);13C NMR(CDCl3，125 MHz)δ:161.2 (C-2)，113.5(C-3)，143.4(C-4)，105.0(C-5)，144.9(C-6)，151.3(C-7)，98.4(C-8)，143.4(C-9)，112.7(C-10)，102.4(11-OCH2O-).The MS and NMR data were consistent with those of ayapin［7］.

7-Hydroxycoumarin(2)White amorphous powder，ESI-MS m/z 163［M+H］+，161［M-H］-.1H NMR (DMSO-d6，500 MHz)δ:10.53(1H，brs，7-OH)，7.92(1H，d，J=9.0 Hz，H-4)，7.52(1H，d，J=8.0 Hz，H-5)，6.79(1H，dd，J=8.0，2.0 Hz，H-6)，6.71 (1H，d，J=2.0 Hz，H-8)，6.20(1H，d，J=9.0 Hz，H-3);13C NMR(DMSO-d6，125 MHz)δ:161.2(C-2)，113.0(C-3)，144.4(C-4)，129.6(C-5)，111.3(C-6)，160.3(C-7)，102.1(C-8)，155.4(C-9)，111.2 (C-10).The MS and NMR data were consistent with those of 7-hydroxycoumarin［8］.

Daphnetin(3) White amorphous powder，ESI-MS m/z 201［M+Na］+，177［M-H］-.1H NMR(CD3OD， 500 MHz)δ:7.85(1H，d，J=10.0 Hz，H-4)，7.01 (1H，d，J=8.0 Hz，H-5)，6.84(1H，d，J=8.0 Hz，H-6)，6.20(1H，d，J=10.0 Hz，H-3);13C NMR (CD3OD，125 MHz)δ:163.5(C-2)，112.1(C-3)，146.7(C-4)，120.2(C-5)，113.9(C-6)，151.1(C-7)，133.5(C-8)，145.0(C-9)，113.8(C-10).The MS and NMR data were consistent with those of daphnetin［9］.

Scopoletin(4) White amorphous powder，ESI-MS m/z 193［M+H］+，215［M+Na］+.1H NMR (CDCl3，500 MHz)δ:7.60(1H，d，J=10.0 Hz，H-4)，6.92(1H，s，H-5)，6.85(1H，s，H-8)，6.27(1H，d，J=10.0 Hz，H-3)，3.95(3H，s，6-OCH3);13C NMR(CDCl3，125 MHz)δ:161.4(C-2)，113.4(C-3)，143.3(C-4)，107.5(C-5)，150.3(C-6)，149.7 (C-7)，103.2(C-8)，144.0(C-9)，111.5(C-10)，56.4(6-OCH3).The MS and NMR data were consistent with those of scopoletin［10］.

Wedelolactone(5) White amorphous powder，ESIMS m/z 651［2M+Na］+，313［M-H］-，627［2MH］-.1H NMR(DMSO-d6，500 MHz)δ:10.80(1H，brs，5-OH)，9.38(2H，brs，11，12-OH)，7.25(1H，s，H-10)，7.17(1H，s，H-13)，6.63(1H，s，H-6)，6.46 (1H，s，H-8)，3.82(3H，s，7-OCH3);13C NMR(DMSO-d6，125 MHz)δ:158.8(C-1)，101.6(C-2)，155.2 (C-3)，96.7(C-4)，154.8(C-5)，93.2(C-6)，162.2 (C-7)，98.1(C-8)，157.7(C-9)，104.5(C-10)，145.4(C-11)，144.3(C-12)，98.8(C-13)，148.8(C-14)，113.7(C-15)，55.7(3H，s，7-OCH3).The MS and NMR data were consistent with those of wedelolactone［11］.

Citrusin C(6) White amorphous powder，ESI-MS m/z 349［M+Na］+，325［M-H］-.1H NMR(CD3OD，500 MHz)δ:7.08(1H，d，J=8.2 Hz，H-6)，6.82 (1H，d，J=1.8 Hz，H-3)，6.72(1H，dd，J=8.2，1.8 Hz，H-5)，5.94(1H，m，H-8)，5.03(2H，m，H-9)，4.84(1H，d，J=7.5 Hz，H-1')，3.86(1H，dd，J= 12.2，1.5 Hz，H-6'a)，3.83(3H，s，2-OCH3)，3.68 (1H，dd，J=12.0，5.0 Hz，H-6'b)，3.48(1H，m，H-2')，3.45(1H，m，H-5')，3.38(2H，m，H-3'，4')，3.33(1H，m，H-7);13C NMR(CD3OD，125 MHz)δ: 146.7(C-1)，151.1(C-2)，114.5(C-3)，136.8(C-4)，122.4(C-5)，118.6(C-6)，41.1(C-7)，139.3(C-8)，116.1(C-9)，103.4(C-1')，75.3(C-2')，78.5 (C-3')，71.7(C-4')，78.2(C-5')，62.8(C-6')，57.0 (2-OCH3).The MS and NMR data were consistent with those of citrusin C［12］.

Medioresinol(7)White amorphous powder，ESI-MS m/z 389［M+H］+，387［M-H］-.1H NMR(CDCl3，500 MHz)δ:6.90(1H，d，J=8.0 Hz，H-5')，6.88 (1H，d，J=2.0 Hz，H-2')，6.82(1H，dd，J=8.0，2.0 Hz，H-6')，6.59(2H，s，H-2，6)，5.59(1H，brs，4'-OH)，5.48(1H，brs，4-OH)，4.75(1H，d，J=4.5 Hz，H-7')，4.72(1H，d，J=4.5 Hz，H-7)，4.26(2H，m，H-9)，3.91(9H，s，3，5，3'-OCH3)，3.90(2H，m，H-9')，3.10(2H，m，H-8，8');13C NMR(CDCl3，125 MHz)δ:134.3(C-1)，102.8(C-2，6)，147.2(C-3，5)，132.9(C-4)，86.2(C-7)，54.4(C-8)，71.9(C-9)，132.2(C-1')，108.6(C-2')，145.3(C-3')，146.7 (C-4')，114.3(C-5')，119.0(C-6')，85.8(C-7')，54.1(C-8')，71.6(C-9')，56.4(3，5-OCH3)，56.0 (3'-OCH3).The MS and NMR data were consistent with those of medioresinol［13］.

Syringic acid(8) Yellow amorphous powder，ESIMS m/z 221［M+Na］+，197［M-H］-.1H NMR(DMSO-d6，500 MHz)δ:12.58(1H，brs，7-COOH)，9.18 (1H，brs，4-OH)，7.21(2H，s，H-2，6)，3.80(6H，s，3，5-OCH3);13C NMR(DMSO-d6，125 MHz)δ:120.4 (C-1)，106.9(C-2，6)，147.4(C-3，5)，140.2(C-4)，167.2(C-7)，56.0(3，5-OCH3).The MS and NMR data were consistent with those of syringic acid［14］.

Vanillic acid(9)White amorphous powder，ESI-MS m/z 167［M-H］-.1H NMR(CD3OD，500 MHz)δ: 7.57(1H，brs，H-2)，7.52(1H，brd，J=8.0 Hz，H-6)，6.80(1H，d，J=8.0 Hz，H-5)，3.89(3H，s，3-OCH3);13C NMR(CD3OD，125 MHz)δ:123.0(C-1)，114.0(C-2)，151.5(C-3)，148.4(C-4)，115.5 (C-5)，124.9(C-6)，170.0(C-7)，56.4(3-OCH3) .The MS and NMR data were consistent with those of vanillic acid［15］.

Isovanillic acid(10)White amorphous powder，ESIMS m/z 167［M-H］-.1H NMR(CD3OD，500 MHz)δ: 7.56(1H，dd，J=8.0，2.0 Hz，H-6)，7.55(1H，d，J= 2.0 Hz，H-2)，6.84(1H，d，J=8.0 Hz，H-5)，3.89 (3H，s，4-OCH3);13C NMR(CD3OD，125 MHz)δ: 123.1(C-1)，115.8(C-2)，148.6(C-3)，152.6(C-4)，113.8(C-5)，125.3(C-6)，170.1(C-7)，56.4(4-OCH3).The MS and NMR data were consistent with those of isovanillic acid［16］.

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