全 文 ：ChemicalconstituentsfromtheleavesofIlexpernyi
XIEGuang-bo1 , NIUFeng3 , WANGXiao-jing2 , LEILian-di1 , TUPeng-fei1, 2＊
(1.SchoolofPharmaceuticalScience, PekingUniversityHealthScienceCenter, Beijing100083 , China;
2.ModernResearchCenterofTraditionalChineseMedicine, PekingUniversity, Beijing100083 , China;
3.ShijiazhuangPharmaGroupNBPPharmaceuticalCo.Ltd, Shijiazhuang052160 , China)
Abstract:Anewcompoundandfiveknowncompoundswereisolatedfromtheethanolicextractofthe
leavesofIlexpernyiFranch.Theirstructureswereestablishedonthebasisofspectralanalysisand
identifiedastrans-isoeugenyl-α-L-arabinopynosyl(1※ 6)-β-D-glucopyranoside(1), kaempferol-3-O-
sambubioside(2), quercetin-3-O-sambubioside(3), isoquercitrin(4), (+)-syringaresinol-O-β-D-
glucopyranoside(5), amarantholidosideIV(6).Amongthem, compound1isanewphenolicglycoside,
namedasilexperphenosideA, andcompounds2-6 wereisolatedfromthisplantforthefirsttime.
Keywords:Ilexpernyi;Ilex;chemicalconstituent;ilexperphenosideA
CLCnumber:R284.1;R284.2　　　Documentcode:A　　　ArticleID:0513-4870(2008)01-0060-03
Received2007-07-19.
ProjectsupportedbyNationalNaturalScienceFoundationofChina
(30672608);ProgramforChangjiangScholarandInnovativeTeam
inUniversity(985-2-063-112).
＊Corespondingauthor　Tel/ Fax:86-10-82802750,
E-mail:pengfeitu@vip.163.com
猫儿刺的化学成分
谢光波 1 , 牛　锋 3 , 王晓静 2 , 雷连娣 1 , 屠鹏飞 1, 2＊
(1.北京大学医学部 药学院 , 北京 100083;2.北京大学 中医药现代研究中心 , 北京 100083;
3.石家庄制药集团恩必普药业有限公司 , 河北 石家庄 052160)
摘要:猫儿刺(Ilexpernyi)为冬青科冬青属植物 ,民间常用于清热解毒 , 润肺止咳。本文对猫儿刺叶的化学成分
进行研究 ,应用硅胶 、SephadexLH-20、反相硅胶和半制备 RP-HPLC等分离纯化方法从其乙醇提取物中分离并鉴定了
6个化合物:(E)-异丁香酚-α-L-吡喃阿拉伯糖基(1※6)-β-D-吡喃葡糖苷(1),山柰酚-3-O-桑布双糖苷(2), 槲皮素-
3-O-桑布双糖苷(3),异槲皮苷(4),丁香树脂酚葡糖苷(5), 反枝苋苷 IV(6)。其中 ,化合物 1为一新酚苷类化合物 ,
命名为猫儿刺酚苷 A。化合物 2 ～ 6为首次从该植物中分离得到。
关键词:猫儿刺;冬青属;化学成分;猫儿刺酚苷 A
　　IlexpernyiFranch.(Aquifoliaceae)isdistributed
mainlyintheYangtzeRivervaleyandsouthregionof
QinlingMountainsinChina.Ithasbeenusedinfolk
medicinetotreattussisandfebris[ 1] .Fewchemical
dataofthisplantareavailablebysurveyingliterature.
Thereforeweinvestigateditschemicalconstituentsand
trans-isoeugenyl-α-L-arabinopynosyl (1※6 )-β-D-
glucopyranoside(1), kaempferol-3-O-sambubioside
(2 )[ 2] , quercetin-3-O-sambubioside (3 )[ 2] ,
isoquercitrin(4)[ 3] , (+)-syringaresinol-O-β-D-glu-
copyranoside(5)[ 4] , amarantholidosideIV (6)[ 5]
wereisolatedfromthisplant.Amongthem, compound
1wasanewphenolicglycoside, namedasilexperphe-
nosideA, andcompounds2-6wereisolatedfromthis
plantforthefirsttime.
Resultsanddiscussion
Compound1(Figure1)wasobtainedascolorless
gum, [ α] 25D -47.1°(c0.7 , CH3OH).TheIR
spectrumof1showedthepresenceofhydroxyl(3 421
cm-1), aromaticring(2 927, 1 511 cm-1 )and
doublebond(1 601, 1 073 cm-1 ) groups.Its
·60· 药学学报 ActaPharmaceuticaSinica2008, 43(1):60-62
DOI :10.16438/j.0513-4870.2008.01.004
molecularformulawasdeterminedtobeC21H30O1 1
byHRESIMS(m/z:481.168 4 [ M+Na] + , calcd
C21H30O11Na, 481.168 0), indicating10 degreesof
unsaturation.The1HNMR spectrum indicatedthe
presenceofABXaromaticprotons[ δ:6.89(1H, dd,
J=2.0, 8.5 Hz), 6.97(1H, d, J=2.0 Hz), 7.09
(1H, d, J=8.5 Hz)] , onemethylgroup[ δ1.83
(3H, dd, J=1.5, 6.5 Hz)] , twoolefinicprotons
[ δ:6.16(1H, dd, J=6.5 , 16.0 Hz), 6.33(1H,
dd, J=2.0, 16.0 Hz)] intrans-configuration, and
onemethoxylgroup[ δ3.85(3H, s)] , suggestingthe
structureofaphenylpropanoidskeletonwithamethoxyl
group, andtwosignalsatδ4.86(1H, d, J=7.0 Hz)
andδ4.27(1H, d, J=7.0 Hz)wereasignedtotwo
anomericprotonsofasugarmoiety.The13CNMR
spectrumconfirmedthepresenceofaphenylpropanoid
moiety. The 13CNMR signalsand the coupling
constant(J=7.0 Hz)oftheanomericprotonsofthe
sugarsuggestedthat1 containedoneβ-D-glucose
moleculeandoneα-L-arabinosemolecule[ 6] .
Figure1　StructureandkeyHMBCcorelationsof
compound1
Thelocationofthetrans-propenylgroupandthe
sugarlinkageincompound1 weredeterminedby2D
NMR spectroscopy.InHMBCspectrum, thelong
rangecorrelationbetweentheolefiniccarbonsignalat
δ131.7 andthearomaticprotonsignalsatδ:6.97
(1H, d, J=2.0Hz), 6.89(1H, dd, J=2.0, 8.5 Hz)
suggestedthatthetrans-propenylgroupwasatachedto
C-4 ofthearomaticring.Meanwhile, thelongrange
couplingbetweentheglucoseanomericprotonδ4.86
(1H, d, J=7.0 Hz)andthearomaticcarbon(δ
146.8, C-1ofphenylpropanoidmoiety), togetherwith
thecorrelationbetweenthearomaticprotonsδ:6.97
(1H, d, J=2.0 Hz), 6.89(1H, dd, J=2.0, 8.5
Hz)andthearomaticcarbon(δ146.8, C-1 of
phenylpropanoidmoiety)inHMBCspectrumsuggested
thatthe sugarmoiety wasatached to C-1 of
phenylpropanoidmoiety.Ontheotherhand, thelong
rangecorrelation between thearabinose anomeric
protonsignalatδ4.27(1H, d, J=7.0 Hz)andthe
carbonsignalatδ69.2(C-6 ofglucose)inHMBC
spectrumindicatedthatthearabinosewasatachedto
C-6 ofglucose.TheD-glucoseandL-arabinosewere
verifiedbyGC analysisfrom theacidhydrolysis
experiment(seeExperimentalpart).Onthebasisof
theabovedata, 1 waselucidated tobe trans-
isoeugenol-1-O-α-L-arabinopynosyl(1※ 6)-β-D-glu-
copyranoside, namedasilexperphenosideA.
Experimental
Generalexperimentalprocedures　NMRdata
weremeasuredonVarianINOVA-500 andVarian
UNITY-500 spectrometerswithtetramethylsilaneasan
internalstandard;A NEXUS-470 FTIR (Nicolet)
spectrophotometerwasusedtorecordtheIRspectrum
(υincm-1 );Opticalrotationwasobtainedona
Perkin-Elmer243Bpolarimeter;HR-ESI-MSspectrum
wasobtainedbyusinganAPEXIIFT-ICRMS(Bruker
Daltonics) spectrometer; ESI-MS spectra were
obtainedbyusingQSTAR(ABI, USA)spectrometer;
GCanalysiswascariedoutonanAgilent6890Ngas
chromatographusingaHP-5 capilarycolumn(28
m ×0.32 mm i.d.):detection, FID;detector
temperature, 260 ℃;columntemperature, 180°;
cariergas, N2;flowrate, 40 mL· min-1;Column
chromatography(CC)wascariedoutbyusingsilica
gel(QingdaoMarineChemicalIndustry, 200 ～ 300
mesh), SephadexLH-20 (Pharmacia), octadecyl
silicagel(ODS, Merck, 25 ～ 40 μm)andD101
porouspolymerresin(TianjinChemicalIndustry);
semi-preparativeHPLC:ODScolumn(300 mm×7.8
mm, 5μm, Waters);ELSDdetector(Altech);flow
rate:2.5 mL· min-1.
Plantmaterial　TheleavesofIlexpernyiwere
colectedbyDr.Si-xiangZhouonApril2005 at
Shennongjia, HubeiProvince, China, andidentified
by Prof. Peng-fei Tu. A voucher specimen
(MEC0504)hasbeendepositedattheherbariumof
Modern Research CenterforTraditionalChinese
Medicine, PekingUniversity.
Extractionandisolation　 Theair-driedand
powderedleaves(15kg)ofIlexpernyiwereextracted
with70% EtOH(90 L)for3 timesat60 ℃.The
EtOHextractwasconcentratedundervacuumandthe
residuewassuspendedinwater, andextractedsucces-
sivelywithpetroleumether, EtOAcandn-BuOH.The
EtOAcextract(230 g)wassubjectedtoCC(SiO2 ,
CHCl3 -MeOH, gradient:from 80∶1 to1∶1)togive
fractions1 ～ 10.Fr.2 wassubjectedtoCC(SiO2 ,
CHCl3-MeOH-H2O 7∶1 ∶0.1 , SephadexLH-20 ,
·61·XIEGuang-bo, etal:ChemicalconstituentsfromtheleavesofIlexpernyi
Table1　1HNMR(500 MHz)and13CNMR(125 MHz)dataofcompound1 inCD3OD
No. δC δH(JinHz) 　　No. δC δH(JinHz)
1 146.8 　- Glc1′ 102.5 4.86(d, 7.0)
2 150.8 　- 2′ 74.8 3.47(m)
3 110.9 6.97(d, 2.0) 3′ 77.6 3.44(m)
4 134.7 　- 4′ 71.5 3.36(m)
5 120.2 6.89(dd, 2.0, 8.5) 5′ 77.4 3.59(m)
6 118.2 7.09(d, 8.5) 6′ 69.2 3.73(m)
4.07(dd, 2.0, 11.5)
7 131.7 6.33(dd, 2.0, 16.0) Ara1″ 104.7 4.27(d, 7.0)
8 125.2 6.16(dd, 6.5, 16.0) 2″ 72.4 3.55(m)
9 18.5 1.83(dd, 1.5, 6.5) 3″ 74.0 3.46(m)
2-OCH3 56.6 3.85(s) 4″ 69.4 3.75(m)
5″ 66.6 3.40(m)
3.80(m)
MeOH)toaford2 (40 mg)and3 (110 mg)
respectively.Fr.7 wassubjectedtoCC(Sephadex
LH-20, CHCl3-MeOH2∶1;SiO2 , CHCl3-MeOH-H2O
5∶1∶0.1)togive4 (30 mg).Then-BuOHextract
(400 g)wasdissolvedinwaterandsubjectedtoCC
(D101 porouspolymerresin), elutingfirstwithH2O,
thenwith10%, 30%, 50%, 70%, 95% EtOHaq.,
separately.Thefractionselutedwith50% and70%
EtOH(97 g)weresubjectedtoCC(SiO2 , CHCl3-
MeOH-H2O, gradient:from10∶1∶0 to1∶1∶0.1)to
givethefractionsa～ k.Fr.bwassubjectedtoCC
(SephadexLH-20, MeOH;ODS, MeOH-H2O2∶3)to
aford5 (20 mg).Fr.dwaspurifiedbyCC
(SephadexLH-20 , MeOH;ODS, MeOH-H2O41∶9,
separately) and then bysemi-preparative HPLC
(MeOH-H2O27∶23)tofurnish6(25mg).Fr.hwas
subjectedtoCC(SiO2 , EtOAc-MeOH-H2O, gradient:
from 60∶1∶0.1 to40∶1∶0.1)andthentosemi-
preparativeHPLC(MeOH-H2O19 ∶31)toaford1
(8mg)。
Identification
IlexperphenosideA(1)　Colorlessgum.[ α] 25D
-47.1°(c0.7, CH3OH);IR(KBr)νmax cm-1:
3 421, 2 927, 1 601, 1 511, 1 263 , 1 073;1HNMR
and13CNMRseeTable1;ESI-MSm/z:476 [ M+
NH4 ] +, 481[ M+Na] +;HR-ESI-MSm/z:481.168 4
[ M+Na] +(calcdC21H30O11Na, 481.168 0).SugaridentificationbyhydrolysisandGC
analysis　Compound1(3mg)washeatedin2 mlof
10% HCl-dioxane(1∶1)at80°for4 h.Afterthe
dioxanewasremoved, thesolutionwasextractedwith
EtOAc(2 mL×3).Theaqueousfractionswere
evaporateandtheresidueconvertedtotheirderivatives
forGCanalysisaccordingtothemethodsdescribedin
theliterature[ 7] .TheretentiontimesofD-glucose(tR,
12.48 min)andL-arabinose(tR, 5.30 min)were
confirmed bycomparisonwith thoseofauthentic
standards[ 8] .
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