全 文 :ChemicalconstituentsofPeriplocaforrestiandtheircytotoxicityactivity
WANGJin1,2,WANGCuifang1,CHENJinxiong1,DUJiang2,QiuDewen2,QIUMinghua1
(1.TheStateKeyLaboratoryofPhytochemistryandPlantResourcesinWestChina,
KunmingInstituteofBotany,ChineseAcademyofSinences,Kunming650204,China;
2.GuiYangColegeofTraditionalChineseMedicine,Guiyang550002,China)
[Abstract] Objective:ToinvestigatethechemicalconstituentsoftherootsofPeriplocaforestiandevaluatetheircytotoxicity
activitiesMethod:SilicagelcolumnchromatographywasemployedfortheisolationandpurificationofchemicalconstituentsThe
structureswereidentifiedonthebasisofspectraldataandthecytotoxicitiesofcompounds24wereinvestigatedbyseveraltumorscel
linesincludingbloodtumor(HL60,CCRTCEM),prostatetumor(PC3,DU145)andMelanoma(UACC62)Result:Fourcom
poundswereisolatedandidentifiedasfolows,lupeol20(29)en3nonadecanoate(1),peroiforosideⅠ (2),3β,5β,14β3OH8β
Hcar20(22)enolide(3),perplocin(4)Conclusion:Compound1isanewlupanetriterpenefatyacidesterCompounds24
showednotablecytotoxicityagainstaltumorlines
[Keywords] Periplocaforesti;lupeol20(29)en3nonadecanoate;peroiforosideⅠ;perplocin;cytotoxicityactivity
[Acceptdate] 20090627
[Foundationitem] TheinnorationdirectionprojectsoftheChinese
academyofScience[KSCX2YWG038(CAS)];Technologysupport
projectforqianfromChineseacademyofScienc(01Qian200501)
[Correspondingauthor] QIUMinghua,Tel:(0871)5223255,Fax:
(0871)5216343,Email:mhchiu@mailkibaccn
PeriplocaforestiSchlechter,aclimbingshrub,is
distributedwidelyinthesouthwestofChinaItsrhizome
hasbeenusedbythepeopleofMiaonationalityforthe
treatmentofrheumatoidarthritis,injuriesfromfals,
gastralia,dyspepsia,amenorhveaandmalaria[1].Pre
viousphytochemicalinvestigationsofthisgenushasled
toisolationofcardenolides,triterpenoidsandsterols[23]
ThetotalglucosidesfromPforestiwerefoundtobe
cardiotonicWhenassayedaccordingtothemethodstat
edfordigitalisintheChinesePharmacopeia,thepigeon
unitofthedrugwas59,10(sd)mg·kg-1[4]The
authorhasinvestigatedthechemicalconstituentsofthis
specias,whichresultedintheisolationofonenewlu
panetriterpanefatyester(1)andthreeknowncarde
nolides(24)Theirstructureswereestablishedaslu
peol20(29)en3nonadecanoate(1),peroiforosideⅠ
(2)[5],3β,5β,14β3OH8βHcar20(22)enolide
(3)[6],andperplocin(4)[7]Thispaperdealswith
theisolationandstructureelucidationofcompounds14
(Fig.1)Thecytotoxicactivitesofcompounds23were
alsoevaluated
Figure1 Thestructuresofcompounds14
1 Generalexperimentalproceduresandplantmaterial
Mp,YANACOMP52apparatusanduncorected
Opticalrotations,HoribaSEAP300spectropolarime
terIRspectra,ShimadzuIR450spectrometerwith
KBrpeletsNMRspecta,BrukerAM400andDRX
500spectrometerswithTMSasaninternalstandard
MS,VGAUTOSPEC3000spectrometer
TherhizomesofPforestiwerepurchasedfrom
theherbalmarketofGuiyang,GuizhouProvince,Chi
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naItwasidentifiedbyProfJiangDuwhoisacoau
thor of this paper, and a voucher specimen
(20030820)wasdepositedattheLaboratoryofPhyto
chemistry,KunmingInstituteofBotany,CAS
2 Extractionandisolation
TheairdriedandmiledrhizomesofPforesti
(10kg)wereextractedwith75% EtOHthreetimes
underrefluxTheethanolicextractwasevaporatedin
vacuotogivetheextract(500g),whichwassuspen
dedinH2O(1500mL),andthenextractedsucces
sivelywithethylacetate(EtOAc)(3×1000mL)and
normalbutanol(nBuOH)(3×1000mL)The
EtOAcextract(250g)wassubjectedtocolumnchro
matography(CC;silicagel;200300mesh)eluting
withCHCl3Me2CO(10∶0,9∶1,8∶2,7∶3,6∶4,0∶
10)togivesixfractionsEachfractionwasfurtherre
peatedlysubjectedtoCC(silicagel;200300mesh)
using a gradientchloroform (CHCl3)methanol
(MeOH)aseluentandpurifiedbySephadexLH20
columnelutedbyMeOHCompounds1(70mg)and
3(76mg)wereafordedfromthefraction1(50g)
Compounds2(80mg)and4(62mg)wereyielded
fromthefraction2(42g)
Compound1(50mg)wasdissolvedinCHCl3(30
mL)andrefluxedwith12mol·L-1NaOH(20mL)
for3hAfterthat,themixturewasneutralizedwith
HCl(01mol·L-1),driedinvacuoandchromato
graphedbycolumnwithsilicagelThefatyacidwas
analyseddirectlybyMSandlupeol(5)wasobtained
byrecrystalizationfromMeOH
3 Resultsanddiscussion
31 Structuralidentification Compound1wasob
tainedaswhitepowder(CHCl3)andanalyzedfor
C39H66O2byHRESIMScombinedwithitsNMRand
DEPTdataItsIRspectrumexhibitedabsorptionbands
forcarbonyl(1730cm-1)andolefinicgroups(1641
and883cm-1)The1HNMRspectrumrevealedthe
presenceofeightmethylgroupsatδH089(3H,s),
091(3H,s),093(3H,s),101(3H,s),105
(3H,s),106(3H,s),110(3H,s)and177
(3H,s),oneprotonofmethineatδH476(1H,dd,
J=9,83Hz),andtwoolefinicprotonsatδH474
and490(1Heach,seach)ItsDEPTspectrumdis
playedeightmethylcarbons,elevenmethylenecarbons
andaseriesofsuccessivemethylenesignalsatca(δC
300,sixmethinesignalsandsixquaternarycarbons
Amongthem,twoolefiniccarbonsatδC1509(quater
narycarbon)and1098(methylene)suggestedthe
presenceofoneisopropenylgroup;onemethinesignal
at(δC806andonequaternarycarbonat(δC1732in
dicatedthepresenceof3βhydroxyesteredAcareful
comparisonofthespectraldataof1withthoseofsome
longchainalkanoicacidestersoflupeol[89] showed
theywereverysimilar,whichrevealedthatcompound
1possessedthesamestructurewithlupanetriterpene
fatyacidesterTheFAB+MSspectrumdisplayedan
[MH]- ionatm/z565consistentwithamolecular
Table1 13C(125MHz)and1HNMR(500MHz)
dataof1(pyridined5)
No δC δH HMBC(HtoC)
1 387(CH2)
2 295(CH2)
3 806(CH) 476(dd,J=9,83Hz) C23
4 382(C)
5 558(CH) 094(dlike)
6 186(CH2) 171(m),177(m)
7 342(CH2)
8 412(C)
9 508(CH) 136(m) C8/C10/C11
10 375(C)
11 213(CH2)
12 256(CH2)
13 385(CH) 175(m) C12/C14/C17/C18
14 432(C)
15 279(CH2) 106(m),100(m)
16 349(CH2) C14/C15/C17/C18/C28
17 433(C)
18 483(CH) 247(m) C13/C17/C28
19 488(CH) 150(m) C18/C20/C21/C29/C30
20 1509(C)
21 294(CH2)
22 403(CH2)
23 282(Me) 106(s) C3/C4/C5/C24
24 169(Me) 093(s) C3/C4/C5/C24
25 164(Me) 101(s) C1/C9/C10/C26
26 163(Me) 105(s) C8/C9/C14/C25
27 148(Me) 110(s) C13/C14/C15
28 182(Me) 091(s) C16/C17/C18/C22
29 1098(CH2) 490(brs),474(brs) C19/C20/C21/C30
30 193(Me) 177(s) C19/C20/C29
1′1732(C)
2′8′300(CH2)
9′ 142(Me) 089(s)
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formulaC39H66O2Onalkalinehydrolysisofthecom
pound1yieldedthecorespondingfatyacidandlupeol
5whichwasconfirmedbycomparisonofitsspectraand
TLCwithauthenticsampleThus,thestructureof1
waselucidatedaslupeol20(29)en3nonadecanoate
Thestructureproposalfor1wasalsoconfirmedbythe
HMBCspectrum(Table1)
32 Cytotoxicassay
Compounds24wereevaluatedfortheircytotoxic
activitiesbybloodtumorcellines(HL60,CCRT
CEM),prostatetumorcellines(PC3,DU145)and
Melanomacelline(UACC62)Compounds24
showedcytotoxicityagainsttoalassayedtumorlines
(Table2),especialycompound4exhibitedsignifi
cantcytotoxicityagainstHL60celswithIC50valuesof
009μmol·L-1;Compoud2showedevidentcytotox
icityagainstHL60andDU145celsTheIC50values
were065and025μmol·L-1,respectively
Table2 Invitrocytotoxicityofcompounds
24(IC50) mol·L
-1
Panel/cellines Histotype 2 3 4
HL60 bloodtumor 0.65 10.05 009
CCRTCEM bloodtumor 2.35 5.27 225
PC3 prostatetumor 1.15 15.28 205
DU145 prostatetumor 0.25 4.18 125
UACC62 melanoma 5.31 11.02 645
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滇杠柳的化学成分及其细胞毒活性筛选
王瑾1,2,王翠芳1,陈进雄1,杜江2,邱德文2,邱明华
(中科院昆明植物研究所 植物化学与西部植物资源持续利用国家重点实验室,云南 昆明 650204;
2.贵阳中医学院,贵州 贵阳 550002)
[摘要] 目的:研究滇杠柳Periplocaforesti的化学成分并筛选其细胞毒活性。方法:利用色谱技术进行分离和纯化,并
通过现代谱学方法鉴定其结构,对分离得到的化合物24用血液系统肿瘤(HL60,CCRTCEM),前列腺肿瘤 (PC3,DU145)
以及黑色素瘤 (UACC62)等细胞系细胞进行细胞毒活性筛选。结果:从滇杠柳中分离鉴定了4个化合物。分别为 upeol20
(29)en3nonadecanoate(1),peroiforoside(2),3β,5β,14β3OH8βHcar20(22)enolide(3),perplocin(4)。结论:其中化
合物1是新化合物,化合物2~4有一定的细胞毒活性。
[关键词] 滇杠柳;lupeol20(29)en3nonadecanoate;peroiforosideⅠ;杠柳苷;细胞毒活性
[责任编辑 王亚君]
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