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PhytochemistryLetters15(2016)1315

ContentslistsavailableatScienceDirect

PhytochemistryLetters
journalhomepage:www.elsevier.com/locate/phytol

Twonewanthraquinoneswithantiviralactivitiesfromthebarksof
Morindacitrifolia(Noni)
JunfengWanga,XiaochuQinb,ZhiyunChena,ZhiranJua,WeijunHea,c,YehuiTana,
XiaojiangZhouc,ZhengchaoTub,FangguoLuc,*,YonghongLiua,*
aCASKeyLaboratoryofTropicalMarineBioresourcesandEcology/GuangdongKeyLaboratoryofMarineMateriaMedica/RNAMCenterforMarine

Microbiology,SouthChinaSeaInstituteofOceanology,ChineseAcademyofSciences,Guangzhou510301,China
bLaboratoryofMolecularEngineeringandLaboratoryofNaturalProductSynthesis,GuangzhouInstitutesofBiomedicineandHealth,ChineseAcademyof
Sciences,Guangzhou510530,China
cSchoolofMedicine,HunanUniversityofChineseMedicine,Changsha410208,China

ARTICLEINFO

ABSTRACT

Articlehistory:
Received1September2015
Receivedinrevisedform31October2015
Accepted9November2015
Availableonline21November2015

Twonewanthraquinones,1,3dihydroxy5methoxy6methoxymethyl2methyl9,10anthraquinone
(1)and1,3dihydroxy5methoxy2,6bismethoxymethyl9,10anthraquinone(2),togetherwithten
knownanthraquinonederivatives(312),threecoumarinderivatives(1315),and6gingerol(16)were
isolatedfromthebarksofMorindacitrifolia(Noni)collectedintheYongxingislandofXisha.The
structuresofcompounds(116)weredeterminedonthebasisofextensivespectroscopicanalyses,as
wellasbycomparisonwithliteraturereports.Thenewcompounds1and2weretestedfortheirantiviral,
cytotoxic,andantibacterialactivities.Intheprimarybioassays,compounds1and2displayedweakanti
H1N1activitywithIC50valuesof66.1and10.5 mM,respectively.Inaddition,compound2showedweak
antiH3N2activitywithIC50valueof11.5 mM,andhadweakantimicrobialactivityagainstStaphylococcus
aureuswithMICvalueof24.5 mM.
2015PhytochemicalSocietyofEurope.PublishedbyElsevierB.V.Allrightsreserved.

Keywords:
Morindacitrifolia
Anthraquinone
Antiviralactivity
Antimicrobialactivity

1.Introduction
MorindacitrifoliaL.,commonlyknownasnoni,belongstothe
Rubiaceaefamily.ThisplantisnativefromSouthEastAsiato
Australia,andhasbeenusedbythePolynesiansformorethan
2000yearsasafood,dye,andherbalremediestocureseveral
diseases(Wangetal.,2002).InSouthEastAsia,thefruitsareused
totreatdiabetes,aswellasfortreatingswollenspleen,liver
diseasesandcough(Eeetal.,2009).Therootsandbarksofnoni
arewellknownfortheirabundanceofanthraquinones(Inoueetal.,
1981Siddiquietal.,2006Eeetal.,2009).Anthraquinonesare
knowntohaveavarietyofinterestingbiologicalprofiles,suchas
antiviral,antioxidant,antimicrobial,cytotoxic,antiinflammatory,
andantiosteoporoticactivities(Akihisaetal.,2007Wuetal.,
2009Locatelli,2011Kremeretal.,2012Locatellietal.,2012
Zhengetal.,
2012Zenginetal.,
2015).Inourcontinuing
investigationsonnovelstructuresfromnoni,twonewanthra
quinones,1,3dihydroxy5methoxy6methoxymethyl2methyl
9,10anthraquinone
(1)
and
1,3dihydroxy5methoxy2,6

*Correspondingauthors.Fax:+8602089023244.
Emailaddresses:lufangguo0731@163.com(F.Lu),yonghongliu@scsio.ac.cn
(Y.Liu).

bismethoxymethyl9,10anthraquinone(2)(Fig.1),togetherwith
tenknownanthraquinonederivatives,1,5,15trimethylmorindol
(3) (Takashimaetal.,2007),morindone6methylether(4)(Ee
etal.,2009),1hydroxy5methoxyanthraquinone(5)(Changand
Lee,1985),1,4dimethoxyl2hydroxyanthraquinone(6)(Lietal.,
2012), 2,3dihydroxyl1methoxylanthraquinone(7)(Lietal.,
2012),rubiadin(8)(Yooetal.,2010),lucidin3methylether(9)
(Fragaetal.,2009),lucidin1,3dimethylether(10)(Fragaetal.,
2009),damnacanthol(11)(Lietal.,2006),digiferruginol(12)
(Zhangetal.,2010),scopoletin(13)(Wuetal.,2009),fraxidin(14)
(Yasudaetal.,2006),isofraxidin(15)(Tianetal.,2008),6gingerol
(16) (Leeetal.,2001)wereisolated.Herein,wereporttheisolation,
structureelucidationandbioactivitiesofthenewcompounds(1
and2)fromthebarksofM.citrifolia.
2.Resultsanddiscussion
Compound(1)wasobtainedasyellowpowderwiththe
molecularformulaC18H16O6asdeterminedbyHRESIMSatm/z
+
329.1016 [M+H] (calcd 329.1020),indicating
11 degreesof
unsaturation.The 1HNMRspectrumcontainedsignalsattributable
tothreearomaticprotons,apairofdoublets(J=7.4Hz)duetotwo
orthopositionedaromaticprotons(H7andH8),and
asinglet

http://dx.doi.org/10.1016/j.phytol.2015.11.006
18743900/2015PhytochemicalSocietyofEurope.PublishedbyElsevierB.V.Allrightsreserved.

14

J.Wangetal./PhytochemistryLetters15(2016)1315

Fig.1.Thechemicalstructuresofthenewcompounds1and2.

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( dH7.16)duetooneisolatedaromaticproton(H4).Thisspectrum
alsoshowedsignalsforfoursubstituents,whichwereassignableto
amethyl,anOmethylgroup,amethoxymethylgroup,anda
13
CNMR
phenolichydroxylperitothecarbonylgroup.The
spectrumof1displayedsignalsforall18Catoms,includingthose
oftheanthraquinoneskeleton( dC185.5,180.9,163.5,162.0,157.9,
140.4,134.2,133.6,133.1,124.8,122.4,116.4,108.3,107.4),one
methylgroup( dC8.0),twoOmethylgroups( dC61.6,58.2),andone
methylenegroup( dC68.3).TheconnectivityoftheprotonsandC
atomswasestablishedbythe 1H,13C,HMQCspectrum(Table1).
TheHMBCcorrelationofCH3( dH2.04)withC1,C2,andC
3suggestedthattheCH3groupshouldbelocatedatC2.The
locationoftheOCH3( dH/C3.81/61.6)atC5wassupportedby
HMBCcorrelationofOCH3( dH3.81)withC5(Fig.2).Thelocation
ofthemethoxymethylgroupatC6wassupportedbyHMBC
correlationofCH2OCH3( dH4.57)withC6(Fig.2).OneOHgroup
locatedatC1wasconfirmedbyHMBCcorrelationsoftheOHH
atomsignal( dH
13.04)withC1,C1a,andC2,indicatinga
phenolicHatomthatishydrogenbondedtotheneighboringCO
group.AnotherphenolicOHgroup,ofwhichtheHatomsignalwas
undetected,shouldbelocatedatC3.Thus,compound1was
identifiedas
1,3dihydroxy5methoxy6methoxymethyl2
methyl9,10anthraquinone.
Compound(2)wasdeducedtohavethemolecularformula
C19H18O7with11degreesofunsaturation,havingoneCH2Ounit
morethan1,onthebasisofHRESIMSdata(m/z359.1114[M+H] + ).
Detailedanalysisofthe1Dand2DNMRspectradatarevealed
that2wasverysimilartothoseof1(Table1),indicatingthatthey
sharedthesameskeleton.However,themethylgroup(2CH3)in
the 1HNMRspectrumof1disappearedinthatof2.In
the 13CNMR

Table1
13
Cand1HNMRspectraldataof1and2(125and500MHz,inDMSOd6, dinppm).
Position

dC Mult
1
1a
2
3
4
4a
5
6
7
8
9
9a
10
10a
11
12
5OCH 3
11OCH 3
12OCH 3
1OH

162.0,s
108.3,s
116.4,s
163.5,s
107.4,d
133.1,s
157.9,s
140.4,s
133.6,d
122.4,d
185.5,s
134.2,s
180.9,s
124.8,s
8.0,q
68.0,t
61.6,q
58.2,q

dH(JinHz)

7.16,s

7.82,d,(7.4)
8.00,d,(7.4)

2.04,s
4.57,s
3.81,s
3.40,s
13.04,s

dC Mult
169.5,s
106.2,s
115.6,s
164.1,s
110.7,d
133.6,s
157.9,s
139.6,s
133.6,d
122.2,d
183.3,s
135.0,s
181.7,s
124.8,s
61.4,t
68.3,t
61.6,q
57.4,q
58.2,q

dH(JinHz)

7.03,s

7.81,d,(7.8)
8.00,d,(7.8)

4.40,s
4.56,s
3.79,m
3.17,s
3.39,s
13.44,s

(DEPT)spectrum,anothermethoxymethylgroupatC2
observedin2,insteadofthemethylgroup(2CH3).Thisdeduction
wasfurthersupportedbytheHMBCcorrelationofH211withC1,
C2,andC3,and11OCH3withC11(Fig.2).Therefore,compound
2wasidentifiedas
1,3dihydroxy5methoxy2,6bismethoxy
methyl9,10anthraquinone.
Thenewcompounds1and2wereevaluatedfortheirantiviral,
antibacterial,andcytotoxicactivities.Intheprimarybioassays,
compounds1and2displayedweakantiH1N1activitywithIC50
valuesof66.1and10.5mM,respectively.Inaddition,compound2
showedweakantiH3N2activitywithIC50valueof11.5mM,and
hadweakantimicrobialactivityagainstStaphylococcusaureuswith
MICvalueof24.5mM.Butnoneofthecompoundsexhibited
cytotoxiceffectsonthetencancercelllines(K562,U937,MOLT4,
HL60,Hela,DU145,MCF7,A549,SGC7901,andH1975).

was

3.Experimental
3.1.Generalexperimentalprocedures
OpticalrotationsweremeasuredwithaPerkineElmer341po
larimeter.UVspectrawererecordedonaShimadzuUV2401PC
spectrometer.1H,13CNMR,DEPT,and2DNMRspectrawere
recordedontheBrukerDRX500spectrometerusingTMSas
internalstandardandchemicalshiftswererecordedasdvalues.
HRESIMS(includingESIMS)spectrawererecordedonanApplied
BiosystemsMariner5140spectrometer.TLCandcolumnchroma
tography(CC)wereperformedonplatesprecoatedwithsilicagel
GF 254(1040mm)andoversilicagel(200300mesh)(Qingdao
MarineChemicalFactory,China),andSephadexLH20(Amersham
Biosciences,Sweden),respectively.Allsolventsusedwereof
analyticalgrade(TianjinFuyuChemicalandIndustryFactory).
SemipreparativeHPLCwasperformedusinganODScolumn(YMC
packODSA,10 250mm,5mm,4mL/min).
3.2.Plantmaterial
ThebarksofM.citrifolia(Noni)werecollectedfromtheXisha
IslandsofChinainNovember2013.Thespecieswasidentifiedby
Dr.ZhiyunChen(oneofourauthors).Andavoucherspecimen(No.
S201301)wasdepositedattheCASKeyLaboratoryofTropical
MarineBioresourcesandEcology,SouthChinaSeaInstituteof
Oceanology,ChineseAcademyofSciences.
3.3.Extractionandisolation
ThedriedandpowderedbarksofM.citrifolia(Noni)(8.5kg)
wereextractedwith80%EtOHunderrefluxfor3
3h.Removalof
thesolventinvacuoaffordedabrownresidue,whichwas
suspendedinH2OfollowedbysuccessivepartitionwithEtOAc
andnBuOH.TheEtOAcextract(63g)wassubjectedtocolumn
chromatography(CC)oversilicagel(200300mesh)elutingwith
gradientCHCl3/MeOH(1:00:1)toaffordsevenfractionsbasedon
TLCproperties.Fraction4waspassedthrough
SephadexLH20

J.Wangetal./PhytochemistryLetters15(2016)1315

15

Fig.2.ThekeyHMBCcorrelationsof1and2.

(MeOH)toobtainsixfractions(Frs.4146).Fr.41waspurifiedby
HPLC(38%CH3CN/H2O)toyield9(5.1mg,tR8.9min),10(6.4mg,tR
14.5min),3(5.3mg,tR17.1min),and16(14.2mg,tR27.7min),
respectively.Fr.45waspurifiedbyHPLC(72%MeOH/H2O)toyield
2 (18.9mg,tR16.9min).Fraction5wasdirectlypurifiedbyHPLC
(75%MeOH/H2O)toyieldthreefractions(Frs.5155),8(12.6mg,
tR16.8min),1(7.1mg,tR18.9min),and4(7.0mg,tR20.1min),
respectively.Fr.51waspurifiedbyHPLC(60%MeOH/H2O)toyield
Fr.511,Fr.512,11(6.8mg,tR8.9min),5(5.1mg,tR12.4min),
and12(9.5mg,tR17.2min),respectively.Fr.511waspurifiedby
HPLC(30%CH3CN/H2O)toyield14(7.4mg,tR
21.8min),13
(19.4mg,tR23.2min),and15(4.3mg,tR26.0min),respectively.
Similarly,Fr.512wasfurtherpurifiedbyHPLC(47%CH3CN/H2O)
toyield
7 (2.4mg,tR
19.2min)and
6 (4.8mg,tR
21.6min),
respectively.
1,3Dihydroxy5methoxy6methoxymethyl2methyl9,10

References
Akihisa,T.,Matsumoto,K.,Tokuda,H.,Yasukawa,K.,Seino,K.,Nakamoto,K.,
Kuninaga,H.,Suzuki,T.,Kimura,Y.,2007.Antiinflammatoryandpotential
cancerchemoprevebtiveconstituentsofthefruitsofMorindacitrifolia(Noni).J.
Nat.Prod.70,754757.
Chang,P.,Lee,K.,1985.Antitumoragents,synthesisofcytotoxicanthraquinones
digiferruginolandmorindaparvinB.J.Nat.Prod.48,948951.
Ee,G.C.L.,Wen,Y.P.,Sukari,M.A.,Go,R.,Lee,H.L.,2009.Anewanthraquinonefrom
Morindacitrifoliaroots.Nat.Prod.Res.23,13221329.
Fraga,B.M.,Quintana,N.,Diaz,C.E.,2009.Anthraquinonesfromnaturaland
transformedrootsofPlocamapendula.Chem.Biodivers.6,182192.
Inoue,K.,Nayeshiro,H.,Inouye,H.,Zenk,M.,1981.Anthraquinonesincell
suspensionculturesofMorindacitrifolia.Phytochemistry20,16931700.
Kremer,D.,Kosalec,I.,Locatelli,M.,Epifano,F.,Genovese,S.,Carlucci,G.,Koncic,M.
Z.,2012.Anthraquinoneprofiles,antioxidantandantimicrobialpropertiesof
Frangularupestris(Scop.)SchurandFrangulaalnusMill.bark.FoodChem.131,
11741180.
Lee,S.W.,Lim,J.,Kim,M.S.,Jeong,J.,Song,G.,Lee,W.S.,Rho,M.,2001.Phenolic
compoundsisolatedfromZingiberofficinalerootsinhibitcelladhesion.Food

http://www.pdf.investintech.com/preview/679baca8100011e694e3002590d31986/index.html

2/3

03/05/2016

www.pdf.investintech.com/preview/679baca8100011e694e3002590d31986/index.html
anthraquinone(1):yellowpowderUV(MeOH) lmax(loge):206
(4.05),248(3.78),280(3.86),410(3.30)nmIR(KBr) nmax3406,
1659,1628,1562,1450,1431,1369,1335,1285,1250,1200,1115,
1069,1011cm11HNMRand13CNMRdata,seeTable1HRESIMS
m/z329.1016[M+H] + (calcdforC18H17O6,329.1020).
1,3Dihydroxy5methoxy2,6bismethoxymethyl9,10an
thraquinone(2):yellowpowderUV(MeOH) lmax(loge):208
(4.11),248(3.98),281(3.94),417(3.44)nmIR(KBr) nmax3395,
1667,1624,1589,1450,1369,1331,1288,1254,1196,1099,1065,
1018cm11HNMRand13CNMRdata,seeTable1HRESIMSm/z
359.1114[M+H] + (calcdforC19H19O7,359.1125).
Acknowledgments
ThisworkwassupportedfinanciallybytheNationalKeyBasic
ResearchProgramofChina(973)sProject(2011CB915503),the
NationalNaturalScienceFoundationofChina(Nos.21502204,
31270402,
21172230,and
41476135),theStrategicPriority
ResearchProgramoftheChineseAcademyofSciences
(XDA11030403),GuangdongMarineEconomicDevelopmentand
InnovationofRegionalDemonstrationProject(GD2012D01001),
theAidprogramforScienceandTechnologyInnovativeResearch
TeaminHigherEducationalInstituionsofHunanProvince(No.15,
Grxjb4).
AppendixA.Supplementarydata
Supplementarydataassociatedwiththisarticlecanbefound,
intheonlineversion,athttp://dx.doi.org/10.1016/j.phytol.
2015.11.006.

Chem.128,778782.
Li,B.,Zhang,D.M.,Luo,Y.M.,Chen,X.G.,2006.Threenewandantitumor
anthraquinoneclycosidesfromLasianthusacuminatissimusMERR.Chem.
Pharm.Bull.54,297300.
Li,Y.,Zheng,C.J.,Qin,L.P.,2012.ChemicalconstituentsfromwholeherbofPaederia
scandensvar.tomentosa.Chin.Tradit.Herb.Drugs43,658660.
Locatelli,M.,2011.Anthraquinones:analyticaltechniquesasanoveltoolto
investigateonthetriggeringofbiologicaltargets.Curr.DrugTargets12,
366380.
Locatelli,M.,Genovese,S.,Carlucci,G.,Kremer,D.,Randic,M.,Epifano,F.,2012.
Developmentandapplicationofhighperformanceliquidchromatographyfor
thestudyoftwonewoxyprenylatedanthraquinonesproducedbyRhamnus
species.J.Chromatogr.A1225,113120.
Siddiqui,B.S.,Sattar,F.A.,Begum,S.,Gulzar,T.,Ahmad,F.,2006.Newanthraquinones
fromthestemofMorindacitrifoliaLinn.Nat.Prod.Res.20,11361144.
Takashima,J.,Ikeda,Y.,Komiyama,K.,Hayashi,M.,Kishida,A.,Ohsaki,A.,2007.New
constituentsfromtheleavesofMorindacitrifolia.Chem.Pharm.Bull.55,
343345.
Tian,F.R.,Zhang,L.,Tian,J.K.,Zhou,W.M.,2008.ChemicalconstituentsofArtemisia
anomalaS.Moore.Chin.J.Med.Chem.18,362365.
Wang,M.Y.,West,B.,Jensen,C.J.,Nowiciki,D.,Su,C.,Palu,A.K.,Anderson,G.,2002.
Morindacitrifolia(Noni):aliteraturereviewandrecentadvancesinnono
research.ActaPharmacol.Sin.23,11271141.
Wu,Y.B.,Zheng,C.J.,Qin,L.P.,Sun,L.N.,Han,T.,Jiao,L.,Zhang,Q.Y.,Wu,J.Z.,2009.
AntiosteoporoticactivityofanthraquinonesfromMorindaofficinalison
osteoblastsandosteoclasts.Molecules14,573583.
Yasuda,T.,Fukui,M.,Nakazawa,T.,Hoshikawa,A.,Ohsawa,K.,2006.Metabolicfate
offraxinadministeredorallytorats.J.Nat.Prod.69,755757.
Yoo,N.H.,Jang,D.S.,Lee,Y.M.,Jeong,H.,Cho,J.,Kim,H.,Kim,J.S.,2010.
AnthraquinonesfromtherootsofKnoxiavalerianoidesinhibittheformationof
advancedglycationendproductsandratlensaldosereductaseinvitro.Arch.
Pharm.Res.33,209214.
Zengin,G.,Locatelli,M.,Ceylan,R.,Aktumsek,A.,2015.Anthraquinoneprofile,
antioxidantandenzymeinhibitoryeffectofrootextractsofeightAsphodeline
taxafromTurkey:canAsphodelinerootsbeconsideredasanewsourceof
naturalcompounds?J.Enzym.Inhib.Med.Chem.doi:http://dx.doi.org/10.3109/
14756366.2015.1063623.
Zhang,H.L.,Zhang,Q.W.,Zhang,X.Q.,Ye,W.C.,Wang,Y.T.,2010.Chemical
constituentsfromtherootsofMorindaofficinalis.Chin.J.Nat.Med.8,192195.
Zheng,C.J.,Shao,C.L.,Guo,Z.Y.,Chen,J.F.,Deng,D.S.,Yang,K.L.,Chen,Y.Y.,Fu,X.M.,
She,Z.G.,Lin,Y.C.,Wang,C.Y.,2012.Bioactivehydroanthraquinonesand
anthraquinonedimmersfromasoftcoralderivedAlternariasp.FungusJ.Nat.
Prod.75,189197.

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