珍珠荚蒾的化学成分研究

李薇薇，史海明，王梦月，李晓波

上海交通大学药学院，上海200240

Introduction

Viburnum foetidum Wall.var.ceanothoides(C.H. Wright)Hand.-Mazz.belongs to the genus Viburnum (Adoxaceae)，distributed in Yunnan，Sichuan and Guizhou provinces of China.The roots，stems，leaves and fruits have been used in Chinese folk medicines to treat stomatitis，eczema，fractures，bone-setters injury and trauma hemorrhage［1］.The phytochemical investigation on V.foetidum var.ceanothoides hasn＇t been reported so far.In our research，fourteen compounds were isolated.This is first report of the presence of compounds 1，7，9，10 and 13 in the genus Viburnum and 1-14 in this plant.

Experimental

General

The MS were measured on an Agilent 1100 series mass spectrometer or Waters UPLC Acquity/QTOFMS Premier.NMR spectra were measured on Bruker DRX-500 or Bruker AvanceⅢ 400 instruments.Shimadzu LC-2010AHT HPLC System and Waters C18column(7.8 ×300 mm，6 μm)were used for semipreparative HPLC.Silica gel for column chromatography and precoated silica GF254plates for TLC were produced by Qingdao Haiyang Chemical Co.Ltd.ODS and D101 macroporous resin were purchased from YMC Co.Ltd，Japan and Tianjin Haiguang Chemical Co.Ltd.respectively.

Plant Material

The stems and leaves of V.foetidum Wall.var. ceanothoides were collected from Chengjiang County，Yunnan Province of China，in May 2008，and were identified by Associate Prof.Wang Meng-Yue.The voucher specimen of V.foetidum var.ceanothoides(No. 080530)was deposited at School of Pharmacy，Shanghai Jiao Tong University.

Extraction and isolation

The dried stems and leaves of V.foetidum var.ceanothoides(23 kg)were milled and extracted three times (3×2 L)with 75%EtOH for 2h each time，with the solvent removed under reduced pressure.The 75%ethanolic extract was suspended in water，and then was partitioned with petroleum ether，CHCl3，EtOAc and n-BuOH successively.The petroleum ether-soluble fraction(90 g)was subjected to silica gel column eluting with a petroleum ether(PE)-EtOAc(100∶0 to 2∶1) gradient system to yield frs.1-8.Fr.4 was chromatographed on a silica gel column eluting with PE-Me2CO (8∶1)to provide frs.4.1-4.5.Fr.4.2 followed by semipreparative HPLC(MeOH-H2O;95∶5)to provide 1(18.2 mg)and 2(19.3 mg).The chloroform-soluble fraction(86 g)was subjected to silica gel column eluting with a PE-EtOAc(100∶0 to 1∶1)gradient system to yield frs.1-10.After recrystallization of fr.3 with CH3OH，3(1.8 g)was obtained.Fr.4 was chromatographed on a silica gel column eluting with PE-Me2CO (6∶1)to afford 4(200 mg).Fr.5 was chromatographed on a silica gel column eluting with PE-Me2CO (5∶1)to afford 5(1.0 g).The EtOAc-soluble fraction (90 g)was subjected to silica gel column eluting with a CH2Cl2-CH3OH(100∶0 to 2∶1)gradient system to yield frs.1-11.Fr.3 was chromatographed on a silica gel column eluting with CH2Cl2-CH3OH(10∶1)to provide frs.3.1-3.4.Fr.3.2 was further purified by semipreparative HPLC(MeCN∶H2O(containing 1‰TFA)=12∶88)to provide 6(6.2 mg)，7(10.6 mg)，8(23.5 mg)and 9(13.5 mg).Fr.4 was purified by preparative TLC with CHCl3-CH3OH-HCOOH (100∶25∶1)，10(24.2 mg)and 11(50.3 mg)were obtained.Fr.6 was applied to silica gel and eluted with CH2Cl2-CH3OH(10∶1)to afford 12(2.0 g).The n-BuOH-soluble fraction(320 g)was subjected to a macroporous resin column with a gradient elution (20%，40%，60%，80%，95%EtOH/H2O)to yield frs.1-5.Fr.2 and Fr.3 was chromatographed on a silica gel column eluting with CH2Cl2-CH3OH(50∶1 to 2∶1)to yield frs.2.1-2.8.Fr.2.3 was subjected to an ODS column eluting with 30%MeOH/H2O to afford Fr.2.3.1-2.3.5.Fr.2.3.3 was purified by preparative TLC with CHCl3-CH3OH-H2O-HCOOH(60∶40∶10∶1)to provide 13(58.2 mg).Fr.2.5 was subjected on silica gel column eluting with CH2Cl2-CH3OH(5∶1)to afford 14(1.2 g).

Identification

Betulin(1)C30H50O2，white powder，ESI-MS m/z: 465.2［M+Na］+;1H NMR(CDCl3，500 MHz)δ: 3.19(1H，dd，J=11.5，4.5 Hz，H-3α)，2.38(1H，m，H-19)，3.34(1H，d，J=11.0 Hz，H-28α)，3.80(1H，dd，J=11.0，1.5 Hz，H-28β)，4.58(1H，brs，H-29α)，4.68(1H，brs，H-29β)，0.97，0.76，0.83，1.02，0.98，1.68(each 3H，s，6×CH3，H-23，24，25，26，27，30);13C NMR(CDCl3，125 MHz)δ:38.6(C-1)，27.3(C-2)，79.0(C-3)，38.8(C-4)，55.2(C-5)，18.3(C-6)，34.2(C-7)，40.9(C-8)，50.3(C-9)，37.1(C-10)，20.8(C-11)，25.1(C-12)，37.2(C-13)，42.7(C-14)，27.0(C-15)，29.1(C-16)，47.7 (C-17)，48.7(C-18)，47.7(C-19)，150.5(C-20)，29.7(C-21)，33.9(C-22)，28.0(C-23)，15.4(C-24)，16.1(C-25)，15.9(C-26)，14.7(C-27)，60.5 (C-28)，109.7(C-29)，19.1(C-30).The NMR spectral data were in consistent with those reported［2］.

Uvaol(2) C30H50O2，white powder，ESI-MS m/z: 465.3［M+Na］+;1H NMR(CDCl3，500 MHz)δ: 5.14(1H，t，J=3.2 Hz，H-12)，3.53(1H，d，J=8.8 Hz，H-28)，3.21(2H，m)，1.11，1.00，0.99，0.95，0.80(each 3H，s，5×CH3)，0.94(3H，d，J=7.2 Hz，H-30)，0.81(3H，d，J=5.0 Hz，H-29);13C NMR (CDCl3，125 MHz)δ:38.7(C-1)，27.2(C-2)，79.0 (C-3)，38.8(C-4)，55.1(C-5)，18.3(C-6)，35.2(C-7)，39.4(C-8)，47.6(C-9)，38.0(C-10)，26.0(C-11)，125.0(C-12)，138.7(C-13)，42.0(C-14)，29.7 (C-15)，23.4(C-16)，36.9(C-17)，54.0(C-18)，39.3(C-19)，40.0(C-20)，32.8(C-21)，30.6(C-22)，28.7(C-23)，15.6(C-24)，15.7(C-25)，17.4 (C-26)，23.3(C-27)，70.0(C-28)，16.8(C-29)， 21.3(C-30).The NMR spectral data were in consistent with those reported［3］.

Betulinic acid(4)C30H48O3，white powder，ESI-MS m/z:455.3［M–H］–;1H NMR(C5D5N，400 MHz) δ:1.82(1H，m，H-2)，3.42(1H，t，J=7.2 Hz，H-3)，2.71(1H，m，H-13)，1.51(1H，m，H-16α)，2.62 (1H，d，J=11.7 Hz，H-16β)，3.51(1H，m，H-19)，4.92(1H，s，H-29α)，4.74(1H，s，H-29β)，0.78，0.98，1.02，1.03，1.19，1.76(each 3H，s，6× CH3);13C NMR(C5D5N，100 MHz)δ:40.5(C-1)，29.6(C-2)，79.3(C-3)，40.8(C-4)，57.2(C-5)，20.0(C-6)，36.1(C-7)，42.3(C-8)，52.2(C-9)，38.8(C-10)，22.4(C-11)，27.4(C-12)，39.8(C-13)，44.1(C-14)，31.5(C-15)，34.1(C-16)，57.9 (C-17)，51.0(C-18)，49.0(C-19)，152.6(C-20)，32.5(C-21)，38.9(C-22)，29.9(C-23)，17.6(C-24)，17.7(C-25)，17.7(C-26)，16.1(C-27)，180.1 (C-28)，111.2(C-29)，20.7(C-30).The NMR spectral data were in consistent with those reported［4］.

p-Hydroxybenzoic acid(6) C7H6O3，white powder，HR-TOF MS m/z:139.0395［M+H］+，137.0239［M–H］–;1H NMR(DMSO-d6，500 MHz)δ:6.80 (2H，d，J=8.0 Hz，H-3，5)，7.78(2H，d，J=8.0 Hz，H-2，6)，10.27(1H，s，OH)，12.40(1H，s，COOH). The NMR spectral data were in consistent with those reported［5］.

4，4'-Dihydroxy-a-truxillic acid(7) C18H16O6，white powder，HR-TOF-MS m/z:327.0847［M –H］–;1H NMR(DMSO-d6，500 MHz)δ:3.65(2H，m，H-β，β＇)，4.12(2H，m，H-α，α＇)，6.69(4H，d，J=8.5 Hz，H-3，3＇，5，5＇)，7.11(4H，d，J=8.5 Hz，H-2，2＇，6，6＇)，9.30(2H，s，2×OH)，11.98(2H，s，2× COOH);13C NMR(DMSO-d6，125 MHz)δ:40.3(C-α，α＇)，46.7(C-β，β＇)，114.9(C-3，3＇，5，5＇)，128.6 (C-2，2＇，6，6＇)，129.6(C-1，1＇)，156.1(C-4，4＇)，173.1(2×COOH).The1H NMR spectral data were in consistent with those reported［6］.

E-p-Coumaric acid(8) C9H8O3，white powder，HR-TOF-MS m/z:165.0552［M+H］+，163.0397［M–H］–。1H NMR(CD3OD，500 MHz)δ:7.60(1H，d，J=16.0 Hz，H-7)，7.44(2H，d，J=8.5 Hz，H-2，6)，6.80(2H，d，J=8.5 Hz，H-3，5)，6.28(1H，d，J= 16.0 Hz，H-8).The NMR spectral data were in consistent with those reported［7］.

Z-p-Coumaric acid(9) C9H8O3，white powder，HR-TOF MS m/z:165.0551［M+H］+，163.0397［M–H］–;1H NMR(CD3OD，500 MHz)δ:7.61(2H，d，J=8.5 Hz，H-2，6)，6.78(1H，d，J=12.5 Hz，H-7)，6.74(2H，d，J=8.5 Hz，H-3，5)，5.78(1H，d，J= 12.5 Hz，H-8).The NMR spectral data were in consistent with those reported［7］.

Phaseic acid(10) C15H20O5，white powder，APCIMS m/z:279.1［M–H］–;1H NMR(CD3OD，500 MHz)δ:1.01(3H，s，H-9＇)，1.21(3H，s，H-7＇)，2.07 (3H，d，J=1.0 Hz，H-6)，2.38(1H，dd，J=18.0，2.5 Hz，H-5＇pro-R)，2.47(1H，dd，J=17.5，2.5 Hz，H-3＇pro-S)，2.70(1H，dd，J=18.0，2.5 Hz，H-5＇pro-S)，2.80(1H，d，J=18.0 Hz，H-3＇pro-R)，3.66(1H，d，J =8.0 Hz，H-8＇pro-S)，3.94(1H，dd，J=8.0，2.5 Hz，H-8＇pro-R)，5.79(1H，brs，H-2)，6.45(1H，d，J =15.5 Hz，H-5)，8.10(1H，d，J=15.5 Hz，H-4);13C NMR(CD3OD，125 MHz)δ:170.0(C-1)，120.2 (C-2)，151.4(C-3)，133.2(C-4)，133.9(C-5)，21.5 (C-6)，83.2(C-1＇)，88.1(C-2＇)，54.3(C-3＇)，211.2 (C-4＇)，53.5(C-5＇)，49.8(C-6＇)，19.7(C-7＇)，78.9 (C-8＇)，16.1(C-9＇).The NMR spectral data were in consistent with those reported［8］.

Protocatechuic acid(11) C7H6O4，white powder，APCI-MS m/z:152.9［M–H］–;1H NMR(CD3OD，500 MHz)δ:6.73(1H，d，J=8.5 Hz，H-5)，7.35 (1H，dd，J=8.5，2.0 Hz，H-6)，7.42(1H，d，J=2.0 Hz，H-2).The NMR spectral data were in consistent with those reported［9］.

1-O-(6-O-α-L-rhamnopyranosyl-β-D-glucopyranosyl)-4-allylbenzene(13) C21H30O10，white powder，ESI-MS m/z:465.2［M+Na］+;1H NMR(CD3OD，500 MHz)δ:1.22(3H，d，J=6.0 Hz，H-6＇＇)，3.33 (3H，m，H-3＇，H-4，H-4＇＇)，3.38(2H，m，H-7)，3.47 (1H，m，H-2＇)，3.55(1H，m，H-5＇)，3.62(1H，dd，J =11.0，6.5 Hz，H-6＇α)，3.68(1H，dd，J=6.0，3.0 Hz，H-5＇＇)，3.72(1H，dd，J=9.5，3.5 Hz，H-3＇＇)，3.86(1H，m，H-2＇＇)，4.03(1H，dd，J=11.0，1.5 Hz，H-6＇β)，4.73(1H，d，J=1.0 Hz，H-1＇＇)，4.82(1H，d，H-1＇)，5.02(1H，m，H-9α)，5.06(1H，m，H-9β)，5.94(1H，m，H-8)，7.03(2H，d，J=8.5 Hz，H-3，5)，7.12(2H，d，J=8.5 Hz，H-2，6);13C NMR(CD3OD， 125 MHz)δ:157.4(C-1)，117.9(C-2)，130.5(C-3)，135.3(C-4)，130.5(C-5)，40.3(C-7)，139.1(C-8)，115.7(C-9)，102.5(C-1＇)，74.9(C-2＇)，78.0 (C-3＇)，71.5(C-4＇)，76.8(C-5＇)，67.8(C-6＇)，102.1(C-1＇＇)，72.4(C-2＇＇)，72.1(C-3＇＇)，74.0(C-4＇＇)，69.8(C-5＇＇)，17.9(C-6＇＇).The NMR spectral data were in consistent with those reported［10］.

Apigenin 7-O-α-L-rhamnopyranosyl(1'''→2'')-β-D-glucopyranoside(14)C27H30O14，yellow needles，ESI-MS m/z:579.2［M+H］+;1H NMR(DMSO-d6，400 MHz)δ:1.20(3H，d，J=6.0 Hz，H-6＇＇＇)，5.13 (brs，H-1＇＇＇)，5.23(1H，d，J=7.5 Hz，H-1＇＇)，6.37 (1H，d，J=2.0 Hz，H-6)，6.79(1H，d，J=2.0 Hz，H-8)，6.88(1H，s，H-3)，6.95(2H，d，J=9.0 Hz，H-3＇，5＇)，7.94(2H，d，J=8.5 Hz，H-2＇，6＇);13C NMR (DMSO-d6，100 MHz)δ:164.2(C-2)，103.2(C-3)，182.0(C-4)，161.4(C-5)，99.3(C-6)，162.5(C-7)，94.5(C-8)，157.0(C-9)，105.4(C-10)，121.0(C-1＇)，128.5(C-2＇，6＇)，116.0(C-3＇，5＇)，161.1(C-4＇)，97.8(C-1＇＇)，76.2(C-2＇＇)，77.2(C-3＇＇)，69.6 (C-4＇＇)，77.0(C-5＇＇)，60.4(C-6＇＇)，100.4(C-1＇＇＇)，70.5(C-2＇＇＇)，70.4(C-3＇＇＇)，71.8(C-4＇＇＇)，68.3(C-5＇＇＇)，18.0(C-6＇＇＇).The NMR spectral data were in consistent with those reported［11］.

β-Sitosterol(3)，ursolic acid(5)and daucosterol (12)were identified by comparison of Rfvalue with the authentic samples.

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