Rev. bras. farmacogn. vol.27 número6

RevistaBrasileiradeFarmacognosia27(2017)785–787

w ww . e l s e v i e r . c o m / l o c a t e / b j p

Short

communication

Chemical

constituents

from

Casearia

spp.

(Flacourtiaceae/Salicaceae

sensu

lato

)

Gerardo

Magela

Vieira

Júnior

,

Luiz

A.

Dutra

,

Roseli

B.

Torres

,

Nivaldo

Boralle

,

Vanderlan

da

S.

Bolzani

_,

_Dulce

_H.

_Siqueira

_Silva

_,

_Mariana

_H.

_Chaves

_,

_Alberto

_J.

_Cavalheiro

a_{DepartamentodeQuímica,CentrodeCiênciasdaNatureza,UniversidadeFederaldoPiauí,Teresina,PI,Brazil}

b_{NúcleodeBioensaio,BiosínteseeEcofisiologiadeProdutosNaturais,InstitutodeQuímica,UniversidadeEstadualPaulista,Araraquara,SP,Brazil} c_{NúcleodoJardimBotânico,InstitutoAgronômicodeCampinas,Campinas,SP,Brazil}

a

r

t

i

c

l

e

i

n

f

o

Articlehistory:

Received26September2017 Accepted5October2017 Availableonline9November2017

Keywords:

Phenoliccompounds Chemicalconstituents Salicaceaesensulato

a

b

s

t

r

a

c

t

ChemicalinvestigationoftheleavesofCaseariagossypiospermaBriq.,Salicaceae,ledtotheisolationof twoknownflavonoids,(+)-taxifolinandquercetin,theleavesofCaseariadecandraJacq.haveafforded hydroquinone,theleavesofCaseariarupestrisEichlerandCasearialasiophyllaEichlerhaveaffordeda diterpene,(E)-phytol,andtheleavesofC.rupestrisandCaseariaobliquaSpreng.haveafforded sitos-terol.ThetwigsofCasearialasiophyllaEichlerledtotheisolationoftwocompounds(+)-pinoresinol,and

N-trans-feruloyltyramine,andthetwigsofC.obliquahaveaffordedN-trans-feruloyltyramine,N-trans -cumaroyltyramine,andcinamicacid.Thisisthefirstreportofthecompounds(+)-taxifolin,quercetin, hydroquinone,(+)-pinoresinolandN-trans-cumaroyltyraminefromtheCaseariagenus.

©2017SociedadeBrasileiradeFarmacognosia.PublishedbyElsevierEditoraLtda.Thisisanopen accessarticleundertheCCBY-NC-NDlicense(http://creativecommons.org/licenses/by-nc-nd/4.0/).

Introduction

Recentphylogeneticandchemical studies,besidesecological

andmorphologicalobservationsfromtheAngiospermPhylogeny

Group (APG) showed that the Flacourtiaceae family was

sepa-ratedintotwoclades(AchariaceaeandSalicaceae),withthegenus

CaseariabelongingtotheSalicaceae(APG,2003).Caseariagenus isfoundinBrazilandcontainsca.of180species,70belongingto theAmericancontinentand37presentinBrazil(Shenetal.,2004).

NochemicalstudieshavebeenpreviouslyreportedforC.

gossy-piospermaBriq.andC.decandraJacq.However,previousstudies ofourgroupwithC.sylvestrisSw.(Santosetal.,2010),C.obliqua

Spreng.(Vieira-Júnioretal.,2009),andC.rupestrisEichler( Vieira-Júnioretal.,2011)related thepresenceofclerodanediterpenes

aswellasinmanyotherCaseariaspecies(Kanokmedhakuletal.,

2007).PreviousstudiesoftheC.lasiopyllaEichlerspeciesdescribed thechemicalcompositionoftheessentialoilsfromleaves(Salvador

etal.,2011).Inthepresentshortcommunication,theoccurrence

ofnineknowncompoundsfromCaseariaspp.isdescribed.

∗ Correspondingauthor.

E-mail:magela@ufpi.edu.br(G.M.VieiraJúnior).

Materialsandmethods

Theaerialparts,twigsandleavesoftheCaseariaspecieswere

collectedinMay2007atCampinasmunicipality,SãoPauloState,

Brazil.Theplantmaterial wasidentified byDr.RoseliB.Torres,

InstitutoAgronômicodeCampinas,Brazil.Avoucherspecimens

[IAC38438(C.gossypiospermaBriq.),IAC42645(C.decandraJacq.), IAC41542(C.rupestris Eichler),IAC42645(C.lasiophyllaEichler),

and IAC46529(C. obliquaSpreng.)] have been deposited in the

herbariumoftheInstitutoAgronômicode Campinas(SãoPaulo

State,Brazil). Opticalrotationwasmeasuredona Perkin-Elmer

341LCpolarimeter.IRspectrawasmeasuredonaNicoletImpact

400spectrometer,usingKBrdisks.The1Dand2DNMR

experi-mentswererecordedonaVarianINOVA500spectrometer(11.7T)

at500MHz(1_H)and125MHz(13_{C)withpulsefieldgradientanda}

VarianINOVA300spectrometer(7.4T)at300MHz(1_H)and75MHz

(13_{C),usingCD3OD,DMSO-d6orCDCl3.Positive-ionHRMSspectra}

wererecordedonanUltrOTOFq(BrukerDaltonics)ESI-qTOFmass

spectrometer,usingTFANaastheinternalstandard.Opencolumn

chromatographywasperformedoversilicagel(40–63␮m,Merck),

silica C18(40␮m, J.T. Baker),or onSephadexLH-20

(Pharma-ciaBiotech).TLCwasperformedusingMercksilicagel60(>230

mesh)andprecoatedsilicagel60PF254plates.SpotsonTLCplates

wereobserved underUV light and byspraying theplates with

anisaldehyde-H2SO4reagent,followedbyheatingat120◦C.All

sol-ventswerepurchasedfromSigma–Aldrich(St.Louis,MO,USA).The

https://doi.org/10.1016/j.bjp.2017.10.003

786 G.M.VieiraJúnioretal./RevistaBrasileiradeFarmacognosia27(2017)785–787

air-driedandpowderedaerialpartsofC.gossypiosperma(leaves

1.4kg),C. decandra(leaves 0.3kg),C. rupestris(leaves0.4kg),C. lasiophylla(leaves0.6kg,andtwigs1.1kg),andC.obliqua(twigs

0.5kg) were exhaustivelyextracted by maceration successively

withhexane,ethanolandwater(3l×3)atroomtemperature.The

aqueousextract(20g) oftheleavesfromC.gossypiospermawas

subjectedtochromatographiccolumnonAmberliteXAD-4using

water, water/MeOH (1:1) and MeOH, to afford (+)-taxifolin (1,

581.0mg, 0.04%– eluted withMeOH).Thehexane extract (4g)

oftheleavesfromC. lasiophyllawaschromatographedover

sil-icagelusinghexanecontainingincreasingamountsofEtOAc.The

subfraction3 (223mg)was subjected to thin-layer preparative

chromatographywithhexane/EtOAc(7:3),resultinginthe

isola-tionof (E)-phytol (13mg,0.002%). Thehexane extract (14g) of

theleavesfromC.rupestriswaschromatographedoversilicagel

usinghexanecontainingincreasingamountsofEtOAc.The

subfrac-tion3(1g)wasrechromatographedinaC-18columnusingwater

containingincreasingamountsofMeOHaffordedamixtureof(E

)-phytol+sitosterol(362mg,0.09%,elutedwithMeOH).Thehexane

extract(1.5g)oftheleavesfromC.obliquawaschromatographed

oversilicagelusinghexanecontainingincreasingamountsofEtOAc

andMeOH. Thesubfraction3(350mg)wasrechromatographed

oversilicagelusinghexanecontainingincreasingamountsofEtOAc

affordedsitosterol(27.2mg, 0.005%), elutedwithhexane/EtOAc

(9:1).Allethanolicextractswere dilutedwithMeOH/H2O(3:1)

andpartitionedwithhexane,Et2O,andEtOAc,successively.The

MeOH–H2Ophase(7.7g)oftheleavesfromC.gossypiospermawas

subjectedtochromatographiccolumnonsilicagelusinghexane

containingincreasingamountsofEtOActoafford(+)-taxifolin(1, 1g,0.07%),elutedwithhexane/EtOAc(1:1).TheEt2Ophase(9.6g)

oftheleavesfromC.gossypiospermawassubjectedtogel

filtra-tion(SephadexLH-20withMeOH)toafford(+)-taxifolin(1,2.8g,

0.2%)andquercetin(37.4mg,0.002%).TheEtOAcphase(6.1g)of

theleavesfromC.decandrawassubjectedtochromatographic

col-umnonsilicagelusinghexanecontainingincreasingamountsof

EtOAc,thesubfraction21(146.1mg)wassubjectedtogelfiltration

(SephadexLH-20withMeOH).Thesubfraction21–4(115mg)was

rechromatographed in EFS-C18 (reverse phase) column using

MeOHaffordedhydroquinone(110.5mg,0.03%).TheEtOAcphase

(6g)ofthetwigsfromC.lasiophyllawassubjectedtogel

filtra-tion(SephadexLH-20withMeOH).Thesubfraction20(332mg)

was chromatographed over silica gel using hexane containing

increasingamountsofEtOActoprovideN-trans-feruloyltyramine

(2,19.5mg,0.001%),elutedwithhexane/EtOAc(3:7).The

subfrac-tion20–10(41mg)wasrechromatographedoversilicagelusing

hexane containingincreasingamounts ofAcOEt toprovide

(+)-pinoresinol(319mg,0.001%),elutedwithhexane/EtOAc(5:5).The EtO2phase(1g)oftheleavesfromC.obliquawassubjectedtogel

filtration(SephadexLH-20withMeOH).Thesubfraction60(32mg)

wassubmitedtopreparativeRP-HPLConaC18columnelutedwith

H2O/MeOH20:80(v/v)ataflowrateof15ml/minandUVdetection

at254nm,affordingthecinnamicacid(9mg,0.001%).The

sub-fraction73 (11mg)wassubmittedtopreparativeRP-HPLCona

C18columneluted withH2O/MeOH/HOAc 33:67:0.01(v/v/v)at

aflow rateof11ml/minand UVdetectionat254nm,affording

theN-trans-cumaroyltyramine(4,2.2mg,0.0004%),andN-trans -feruloyltyramine(2,4.5mg,0.0009%).

Resultsanddiscussion

Thestructuresoftheninecompoundswereidentifiedby

com-parisonoftheiropticalrotation,spectroscopicandspectrometric

data(UV,IR,HRESIMSorESIMSand1_Hand13_{CNMR)withthose}

reportedintheliterature(seeSupportingInformation).

Thisworkreportedtheisolationoftwoflavonoids((+)-taxifolin

(1)andquercetin),onesimplephenoliccompound(hydroquinone),

one diterpene ((E)-phytol), one steroid (sitosterol), one lignan

((+)-pinoresinol (3)), two amides (N-trans-feruloyltyramine (2) andN-trans-cumaroyltyramine(4)),andcinnamicacidfromfive

Caseariaspecies.Allcompoundswereisolatedfromthesespecies

for the first time. Except for (E)-phytol, sitosterol, N-trans

G.M.VieiraJúnioretal./RevistaBrasileiradeFarmacognosia27(2017)785–787 787

beenisolatedpreviouslyfromC.grewiifolia(Rayaniletal.,2012),C. sylvestris(Wangetal.,2009),C.membranacea(Changetal.,2003), andC.graveolens(Talapatraetal.,1983),N-trans-feruloyltyramine (2)fromC. membranacea(Chang etal., 2003)and C. grewiifolia

(Rayanil etal., 2012), (E)-phytol from C.balansae (Wanget al., 2013),and cinnamic acidfromC. sylvestris(Wanget al.,2009).

Caseariaspeciescontainsflavonoids(Raslanetal.,2002),lignans (Wangetal.,2010),steroids(Wangetal.,2009;Changetal.,2003), coumarins(Talapatraetal.,1983),amides(Changetal.,2003), ter-penoids(Bouetal.,2014),phenoliccompounds(Chaietal.,2010), andmainlyclerodanediterpenes(Vieira-Júnioretal.,2009; Vieira-Júnioretal.,2011;Bouetal.,2014).

The compound (+)-taxifolin (1) wasisolated in a significant

amount(3.8g)fromleavesofC.gossypiosperma,butitcanbefound inlargerquantitiesinconifers,e.g.,theSiberianlarch,Larixsibirica,

PinusroxburghiiandCedrusdeodara(Willföretal.,2009).The

clero-danediterpenesareconsideredchemosystematicmarkersofthis

genus,buttheliteraturedoesnotdescribethepresenceofthese

substancesinC.lasiophylla,C.gossypiospermaandC.decandra. Thebestofourknowledge,thisisthefirstreportofthechemical

constituentsofC.gossypiospermaandC.decandra.Thecompounds

(+)-taxifolin(1),quercetin,hydroquinone,(+)-pinoresinol(3)and

N-trans-cumaroyltyramine(4)arereportedforthefirsttimeinthe

Caseariagenus.From1.4kgofdriedleavesfromC.gossypiosperma, anamountof3.8gof(+)-taxifolin(1)wasobtained.Thisextract

isa rich sourceof(+)-taxifolin(1), which mayofferinteresting

potentialapplicationsinthefood,cosmeticand/orpharmaceutical

industries.

Authorcontributions

LADandGMVJcarriedoutlaboratoryworkaspartofherfinal

yearresearchproject. NBandGMVJ obtainedtheNMRand MS

dataandcontributedtocompound identification.RBTidentified

and deposited thespecimens in the herbariumof theInstituto

AgronômicodeCampinas(SãoPauloState,Brazil).GMVJ,MHC,VSB,

DHSS,andAJCsupervisedthisproject,providedintellectualinput

andpreparedthemanuscript.Alltheauthorshavereadthefinal

manuscriptandapprovedthesubmission.

Conflictsofinterest

Theauthorsdeclarenoconflictsofinterest.

Acknowledgments

TheauthorsaregratefultoFundac¸ãodeAmparoàPesquisado

EstadodeSãoPauloandBiotaInstitutoVirtualdaBiodiversidade

forfinancialsupportoftheProjectno.03/02176-7.Wearegrateful

theProf.Dr.NorbertoPeporineLopesforuseoftheLC–MS.

AppendixA. Supplementarydata

Supplementarydataassociatedwiththisarticlecanbefound,in theonlineversion,atdoi:10.1016/j.bjp.2017.10.003.

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