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Chemical constituents from Mylabris phalerata and their cytotoxic activity in vitro

斑蝥化学成分及体外抗肿瘤活性研究



全 文 :2016
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Vol41,No.5 March,2016
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] 20151028

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(cstc2014yykfC10004)

yzA{
] 
!?¸
,Tex/Fax:(023)89029081,Email:yangdaji@hotmailcom;
ea
,Tex/Fax:(023)89029070,Email:cqzhangyi@hot
mailcom

A{„…
] 
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Οqré
,Email:zengyaobo@126com
ChemicalconstituentsfromMylabrisphalerataand
theircytotoxicactivityinvitro
ZENGYaobo1,LIUXiaoling1,LIChuangjun2,ZHOUXing1,ZHANGXiaomei1,YANGYong1,
DUHongfei1,TANChunbin1,ZHANGYuyu1,ZHANGYi1,YANGDajian1
(1ChongqingAcademyofChineseMaterialMedical,Chongqing400065,China;
2StateKeyLaboratoryofBioactiveSubstanceandFunctionofNaturalMedicine,InstituteofMateriaMedica,
ChineseAcademyofMedicalSciences,PekingUnionMedicalColege,Beijing100050,China)
[Abstact] TencompoundswereisolatedfromMylabrisphaleratabyusingpreparativeHPLCandcolumnchromatographyoverMCI
gelOnthebasisofphysicalchemicalproperties,NMRandMSdataanalysis,thecompoundswereidentifiedas5′[(1R,2R,3S,
6R)1hydroxymethyl2methyl3,6epoxycyclohexane1,2dicarboximide]ethyl2′methyl2′butenoate(1),cantharidin(2),cyclo
(LProLAla)(3),cyclo(RProRLeu)(4),cyclo(SProRLeu)(5),cyclo(DProLTyr)(6),indole3aldehyde(7),3
indoleaceticacid(8),valerolactam(9),and4hydroxyphthalid(10)Compound1wasanewcompound,andcompounds210were
obtainedfromthisgenusforthefirsttimeCompounds19weresubjectedtocytotoxicactivityonHCT116,HepG2,BGC823,NCI
H1650,A2780cellines,andonlycompound2showedinhibitoryefectonalcancercellines
[Keywords] Mylabris;chemicalconstituents;cytotoxicefect
doi:10.4268/cjcmm20160516
·958·
2016
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Vol41,No.5 March,2016
  
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Table1 1HNMRand13CNMRspectraldataforcompound1(inCD3OD)
No δH δC No δH δC
1 597 10 420(1H,d,J=116Hz) 611
2 553 447(1H,d,J=116Hz)
3 453(1H,d,J=4Hz) 859 1′ 1673
4 170(1H,m),177(1H,m) 248 2′ 1604
5 171(1H,m),197(1H,m) 256 3′ 562(1H,m) 1161
6 460(1H,d,J=4Hz) 833 4′ 189(3H,d,J=18Hz) 277
7 1806 5′ 360(2H,d,J=16Hz) 425
9 1832 6′ 360(2H,d,J=16Hz) 596
2Me 125(3H,s) 126 2′Me 213(3H,d,J=12Hz) 207
  
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1HNMR(CDCl3,400MHz)δ:124(6H,s,1Me,2
Me),176(2H,m,H4,H5),181(2H,m,H4,H
5),472(2H,brs,H3,H6);13CNMR(CDCl3,125
MHz)δ:127(1Me,2Me),234(C4,C5),552
(C1,C2),847(C3,C6),1759(C7,C9)。
2
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3 
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Hz,9Me),193(1H,m,H4a),200(1H,m,H
4b),202(1H,m,H5a),231(1H,m,H5b),352
(2H,m,H3),425(1H,t,J=84Hz,H6),418
(1H,q,J=64Hz,H9);13CNMR(CD3OD,125
MHz)δ:157(9Me),236(C4),291(C5),464
(C3),521(C9),605(C6),1690(C7),1726
(C1)。
2¼=‰ª^•
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4 
·>%¥

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1HNMR(CD3OD,400MHz)δ:101(6H,dd,J=
68,52Hz,2Me,13Me),195(1H,m,H4a),
199(1H,m,H4b),204(1H,m,H5a),237(1
H,m,H5b),352(2H,m,H3),386(1H,ddd,
J=92,52,40Hz,H9),428(1H,t,J=80Hz,
H6),159(1H,m,H10a),168(1H,m,H11),
180(1H,m,H10b);13CNMR(CD3OD,125MHz)
δ:205(12Me),216(13Me),219(C4),241(C
11),285(C5),422(C10),455(C3),557(C
9),579(C6),1676(C7),1702(C1)。

[10]
kBŽ=‰Ë§P+±¹ª2¼=‰£4[
Q

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1H,13CNMR
!’ñŸç=‰
·168·
2016
y

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41
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Vol41,No.5 March,2016
Ò×µ_ñ.¯
cyclo(RProRLeu)。
_ñ.
5 
·>%¥

)z
),[α]25D =-89
(c008,MeOH);ESIMSm/z2332[M+Na]+;
1HNMR(CDCl3,400MHz)δ:095(3H,d,J=64
Hz,13Me),099(3H,d,J=64Hz,12Me),190
(1H,m,H4a),208(1H,m,H4b),214(1H,m,H
5a),234(1H,m,H5b),353(1H,dt,J=84,36
Hz,H3b),360(1H,dt,J=84,36Hz,H3a),
399(1H,d,J=92,H9),410(1H,t,J=80Hz,
H6),612(1H,s,NH),152(1H,m,H10a),172
(1H,m,H11),193(1H,m,H10b);13CNMR
(CDCl3,125MHz)δ:212(12Me),228(13Me),
233(C4),247(C11),282(C5),386(C10),
455(C3),535(C9),590(C6),1662(C7),
1703(C1)。
2¼=‰ª^•
[10]
kBŽuç=‰
˧P+±¹[Q

ÝÒ×µ_ñ.¯
cyclo(S
ProRLeu)。
_ñ.
6 
„Cv‹.

ëÐ
),[α]25D =-51
(c03,MeOH);ESIMSm/z2832[M+Na]+;
1HNMR(CD3OD,400MHz)δ:123(1H,m,H4a),
181(2H,m,H4b,H5a),210(1H,m,H5b),308
(2H,m,H10),359(2H,m,H3),405(1H,m,
H9),437(1H,brs,H6),671(2H,d,J=64Hz,
H3′,H5′),703(2H,d,J=68Hz,H2′,H6′);
13CNMR(125MHz,CD3OD)δ:227(C4),294
(C5),377(C10),459(C3),579(C9),600
(C6),1162(C3′,C5′),1321(C1′),1275
(C2′,C6′),1578(C4′),1670(C7),1708
(C1)。
2¼=‰ª^•
[11]
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ÝÒ×µ_ñ.¯
cyclo(DProLTyr)。
_ñ.
7 
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ëÐ
),ESIMSm/z
1681[M+Na]+;1HNMR(CD3OD,400MHz)δ:
727(2H,m,H6,H7),748(1H,d,J=80Hz,H
8),811(1H,s,H2),818(1H,d,J=76Hz,H
5),990(1H,s,H10);13CNMR(CD3OD,125MHz)
δ:1134(C8),1204(C3),1227(C7),1239
(C6),1253(C5),1260(C4),1392(C9),
1400(C2),1874(C10)。
2¼uç=‰ª^•
[12]
kBŽ=‰[Q

ÝÒ×µ_ñ.¯
indole3
aldehyde。
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8 
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1971[M+Na]+;1HNMR(CD3OD,400MHz)δ:
754(1H,d,J=80Hz,H4),735(1H,d,J=80
Hz,H7),718(1H,s,H2),711(1H,t,J=72Hz,
H6),702(1H,t,J=72Hz,H5),365(2H,s,
H8);13CNMR(CD3OD,125MHz)δ:1193(C2),
1285(C3),1095(C3a),1200(C4),1226
(C5),1249(C6),1123(C7),1381(C7a),
335(C8),1780(C9)。
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kBŽ=‰[Q

ÝÒ×µ_ñ.¯
3indolea
ceticacid。
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1222[M+Na]+;1HNMR(CD3OD,400MHz)δ:
179(2H,m,H5),181(2H,m,H4),233(2H,t,
H3),332(2H,m,H6),591(1H,s,NH);
13CNMR(CD3OD,125MHz)δ:1722(C2),424
(C6),315(C3),223(C5),209(C4)。
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ÝÒ×µ_ñ.¯
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J=76Hz,H6),708(1H,d,J=80Hz,H4),
530(2H,s,H3);13CNMR(CD3OD,400MHz)δ:
700(C7),1171(C4),1214(C6),1285(C9),
1320(C5),1351(C8),1542(C3),1741(C
2)。
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[1
[S]2015
[2] HuhJE,KangKS,ChaeC,etalRolesofp38andJNKmito
genactivatedproteinkinasepathwaysduringcantharidininduced
apoptosisinU937cels[J].BiochemPharmacol,2004,67:
1811
[3] PengF,WeiYQ,TianL,etalInductionofapoptosisbynor
cantharidininhumancolorectalcarcinomacellines:involvement
oftheCD95receptor/ligand[J].JCancerResClinOncol,
2002,128:223
[4] ChenYN,ChenJC,YinSC,etalEfectormechanismsof
norcantharidininducedmitoticarestandapoptosisinhuman
hepatomacels[J].IntJCancer,2002,100:158
[5] HuanSK,LeeHH,LiuDZ,etalCantharidininducedcyto
toxicityandcyclooxygenase2expressioninhumanbladdercarci
nomacelline[J].Toxicology,2006,223:136
[6] WiliamsLA,MolerW,MerisorE,etalInvitroantiprolifer
ation/cytotoxicactivityofcantharidin(SpanishFly)andrelated
derivatives[J].WestIndianMedJ,2003,52:10
[7] NakataniT,JinpoK,NodaNCantharimideDimersfromthe
ChineseBlisterBeetle,MylabrisphaleratePALLAS[J].Chem
PharmBul,2007,55(1):92
[8] VermaAK,PrasadSBChangesinglutathione,oxidativestress
andmitochondrialmembranepotentialinapoptosisinvolvingthe
anticanceractivityofcantharidinisolatedfromredheadedblister
beetles,epicautahirticornis[J].AntiCancerAgentMedChem,
2013,13:1096
[9] FrancisJS,DavidJV,KeithHH,etalMetabolitesfromthe
marinespongeTedaniaignisAnewatisanediolandseveral
knowndiketopiperazines[J].JOrgChem,1983,48:3941
[10] AdamczeskiM,ReedAR,CrewsPNewandknowndike
topiperazinesfrom thecaribbeansponge,Calyxcf.podatypa
[J].JNatProd,1995,58(2):201
[11] 
±º‡

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âŽÕÌ8©XÀÖ¬–•
„Ð{-1X_‡˜™
[J].
$Dwk

ãä&
æ
,2014,53(2):78
[12] NakajimaE,NakanoH,YamadaK,etalIsolationandidentifi
cationoflateralbudgrowthinhibitor,indole3aldehyde,in
volvedinapicaldominanceofpeaseedlings[J]Phytochemis
try,2002,61:863
[13] SunDD,DongW W,LiX,etalIndolealkaloidsfromthe
rootsofIsatisingigoticaandtheirantiherpessimplexvirustype2
(HSV2)activityinvitro[J]ChemNatCompd,2010,46
(5):763
[14] LiL,YanYM,LiXN,etalNitrogenouscompoundsfromHo
lotrichiadiomphaliaBates[J].NatProdResDev,2013,25:
329
[15] 
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HH[<_‡ËÌöÖ
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[J].
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