Synthesis of New Hydrated Geranylphenols and in Vitro Antifungal Activity against Botrytis cinerea

18 Pages·2016·2.71 MB·English

International Journal o f
Molecular Sciences
Article
Synthesis of New Hydrated Geranylphenols and
in Vitro Antifungal Activity against Botrytis cinerea
MauricioSoto1,LuisEspinoza1,MaríaI.Chávez1,KatyDíaz1,AndrésF.Olea2
andLautaroTaborga1,*
1 DepartamentodeQuímica,UniversidadTécnicaFedericoSantaMaría,Valparaíso2340000,Chile;
[email protected](M.S.);[email protected](L.E.);[email protected](M.I.C.);
[email protected](K.D.)
2 InstitutodeCienciasQuímicasAplicadas,FacultaddeIngeniería,UniversidadAutónomadeChile,
Santiago8910339,Chile;[email protected]
* Correspondence:[email protected];Tel.:[phone]
AcademicEditor:ZdenekWimmer
Received:18April2016;Accepted:25May2016;Published:3June2016
Abstract: GeranylatedhydroquinonesandothergeranylatedcompoundsisolatedfromAplydium
species have shown interesting biological activities. This fact has prompted a number of studies
wheregeranylatedphenolderivativeshavebeensynthesizedinordertoassaytheirbioactivities. In
thiswork,wereportthesynthesisofaseriesofnewhydratedgeranylphenolsusingtwodifferent
syntheticapproachesandtheirinhibitoryeffectsonthemycelialgrowthofBotrytiscinerea. Fivenew
hydratedgeranylphenolswereobtainedbydirectcouplingreactionbetweengeraniolandphenol
in dioxane/water and using BF ¨Et O as the catalyst or by the reaction of a geranylated phenol
3 2
withBF ¨Et O.Twonewgeranylatedquinoneswerealsoobtained. Thesynthesisandstructural
3 2
elucidationofallnewcompoundsispresented. Allhydratedgeranylphenolsefficientlyinhibitthe
mycelialgrowthofB.cinerea.Theiractivityishigherthanthatobservedfornon-hydratedcompounds.
Theseresultsindicatethatstructuralmodificationonthegeranylchainbringsaboutanenhancement
oftheinhibitioneffectofgeranylatedphenolderivatives.
Keywords: geranylated phenol derivatives; hydrated geranyl; synthesis; structural elucidation;
growthinhibitioneffect;Botrytiscinerea;fungicide
1. Introduction
Thesubgroupsoflineargeranylatedquinones,geranylatedhydroquinonesandmeroterpenes
are represented by an important number of metabolites isolated from ascidians belonging to the
genusAplydium[1,2]. Thefirstbiologicallyactivetunicatemetaboliteswere2-geranylhydroquinone
(1) and 2-geranyl hydroquinone diacetate (2) (Figure 1), isolated from Aplydium sp. and
Phaceliacrenulata[3,4]andPyrolajaponica[5],respectively,andlaterfoundinmanyothersAplydium
species. Ithasbeenshownthatthesecompoundsexhibitantitumoralactivity[6]. Additionally,several
linearquinones/hydroquinonescarryingageranyltypesidechain(Compounds1,3–11; Figure1)
havebeenobtainedfromdiverseAplydiumspecies[7–12]. Compound1and2-geranylbenzoquinone
(3)exhibitinterestingandvariousbiologicalactivities[3,8,13–17],whereasCompounds5and6show
antioxidantactivities[8],andCompound4showscytotoxicityactivityagainstP-388mouselymphoma
suspensionculture[18]. Additionally,lineargeranylmethoxyphenol/acetatesderivativesisolatedfrom
Phaceliaixodes[19]arecytotoxic,allergenicandinsecticidal.
Ontheotherhand,theanticancerproperties,bothinvitroandinvivo,ofagroupofprenylated
quinones,i.e.,3-demethylubiquinoneQ2(9)anditssyntheticanalogs,werestudiedasafunctionof
theirmolecularstructure[20,21]. Theresultsindicatethat9anditsderivativesareabletoinhibitthe
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growth of human cancer cell lines and of the solid Ehrlich carcinoma in mice by inducing apoptosis
ginr ocwanthceorf cheullms. aTnhec aanncteicracnecllelri naectsivanitdy ooff tthheiss sotlriudcEtuhrralilc-rheclaatrecdin foammailiyn omf cicoembpyoiunnddusc idnegpaepnodpst oosni sthine
cleanngctehr ocef ltlhse. Tphoelyapnrteincyanl ccheraianc tainvdit yono fththei psostsriuticotnu oraf lt-hreel amteedthfoaxmyli lgyroofucposm inp tohuen qdusindoenpee npdasrto onf tthhee
lmenogletchuolef t[h20e]p. Tolhyep mreonsytl ecfhfeacitnivaen cdoomnptohuenpdoss witieorne othfothsee hmaevtihnogx ay sligdreo uchpasinin otfh teheq ugienroanneylp tayrpteo (fttwhoe
misooplerceunlee u[2n0it]s.)T ahnedm oonset meffeetchtoivxeylc ogmropuopu antd tshew pearerat-hpoosseithioavni tnog tahes igdeercahnayiln cohfaitnh.e geranyltype(two
isoprCenyecluondiitpsr)eannydl ohnyedmroeqtuhionxoynlegsr,o usupcaht tahse mpaertah-opxoyscitoinoindtioolt h(e12g)e raanndy lccohnaiitnri.ol (13), have been
isolatCeydc florodmip rAenplyidlihuymd raoffq. udiennosnuems ,[s2u2c] hanasdm Aeptlhidoixuymc ocnonidicioulm(1 [29)],a rnedspcoecntiitvrieolly(. 1M3)e,thhaovxeybceoennidisiooll aatnedd
fitrso mmeAtphloidxiyu mdearffi.vdaetnivseu mw[e2r2e] saynndthAepsliizdieudm, acnondi ctuhmei[r9 b],iorelospgeiccatilv aeclyti.vMitieetsh ooxny chounmidaino lcaanndceitrs cmelelt hlionxeys
danerdi vsaetaiv uerwcheirne esymnbtrhyeossiz ewde,raen adssthesesiredb i[o2l3o]g. icAa ldaecttaivilietdie sdoenschriupmtioann coafn tcheer icseolllalitnioens aanndd sbeiaoluorgcihcianl
eamctibvriytioess wofe rteheassese asnsedd o[t2h3e]r. Astrduecttauilreeds dofe sncartiputriaoln porfenthyelqisuoilnaotinoens,a nhdydbriooqlougiincoanlaecst aivnidti emseorfothteerspeeanneds
ocathne bres tfrouucntudr iens [o2f4n,2a5tu].r alprenylquinones,hydroquinonesandmeroterpenescanbefoundin[24,25].

FFiigguurree 11.. SSttrruucctuturreeo offs osmomene antuartualrlayl-loyc-coucrcruinrrginlign elianregaerr agneyralantyeldatheydd rhoyqdurionqounienso/nqeusi/nqounienso1n–e1s1 1,–a1n1d,
cayncdl ocdyicplorednipyrlemneyrlo mteerrpoetnerepse1n2e–s1 31.2–13.
Thus, the interesting biological activity shown by 1 and other prenyl derivatives [17] (see
Thus,theinterestingbiologicalactivityshownby1andotherprenylderivatives[17](seeFigure1)
Figure 1) has prompted us to undertake the synthesis of a significant number of linear geranylated
haspromptedustoundertakethesynthesisofasignificantnumberoflineargeranylatedphenols,
phenols, including Compounds 1–3 and some geranylated methoxyphenyl/acetate analogs [26–32],
includingCompounds1–3andsomegeranylatedmethoxyphenyl/acetateanalogs[26–32],inorderto
in order to evaluate the in vitro cytotoxic activity on some cancer lines and the inhibitory effects on
evaluatetheinvitrocytotoxicactivityonsomecancerlinesandtheinhibitoryeffectsonthemycelial
the mycelial growth of plant pathogen Botrytis cinerea [29–31]. The latter is a facultative
growthofplantpathogenBotrytiscinerea[29–31]. Thelatterisafacultativephytopathogenicfungus
phytopathogenic fungus that attacks the flowers, fruits, leaves and stems of more than 200 plant
thatattackstheflowers,fruits,leavesandstemsofmorethan200plantspecies[33]. InChile,there
species [33]. In Chile, there is a high incidence of this fungus, and its control by commercial fungicides
is a high incidence of this fungus, and its control by commercial fungicides (dicarboximides and
(dicarboximides and benzimidazoles) is becoming more ineffective due to the appearance of highly
benzimidazoles)isbecomingmoreineffectiveduetotheappearanceofhighlyresistantstrains[34,35].
resistant strains [34,35]. Thus, an increasing number of metabolites isolated from plants,
Thus,anincreasingnumberofmetabolitesisolatedfromplants,hemisyntheticandsyntheticproducts
hemisynthetic and synthetic products have been studied as an alternative to chemical
havebeenstudiedasanalternativetochemicalfungicide[29,36–38]. Previousworkhasshownthat
fungicide [29,36–38]. Previous work has shown that the anti-fungal activity of geranylated phenols is
theanti-fungalactivityofgeranylatedphenolsismainlydeterminedbythepresenceofthegeranyl
mainly determined by the presence of the geranyl chain and substitution on the aromatic
chainandsubstitutiononthearomaticring[29,31]. However,therearenodataregardingthefungicide
ring [29,31]. However, there are no data regarding the fungicide activity of compounds carrying a
activityofcompoundscarryingamodifiedgeranylchain,e.g.,hydratedgeranylphenols.
modified geranyl chain, e.g., hydrated geranylphenols.
Therefore, in this research, a study of the inhibitory effects on the mycelial growth of plant
Therefore, in this research, a study of the inhibitory effects on the mycelial growth of plant
pathogenB.cinereaofgeranylatedphenols(Compounds14–18),geranylatedquinones(Compounds
pathogen B. cinerea of geranylated phenols (Compounds 14–18), geranylated quinones (Compounds
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Int. J. Mol. Sci. 2016, 17, 840 3 of 18
1199––2211)) aanndd hhyyddrraatteedd ggeerraannyyllpphheennoollss ddeerriivvaattiivveess ((CCoommppoouunnddss 2222––2266)) ((sseeee FFiigguurree 22)) iiss rreeppoorrtteedd.. TThhee
ssyynntthheessiiss aanndd ssttrruuccttuurraall eelluucciiddaattiioonn ooff tthhee nneeww ccoommppoouunnddss ((1144––1188,, 2200,, 2222––2266)) iiss aallssoo pprreesseenntteedd..

FFiigguurree 22.. CChheemmiiccaall ssttrruuccttuurreess ooff ggeerraannyyllaatteedd pphheennoollss ((1144––1188)) ggeerraannyyllaatteedd qquuiinnoonneess ((1199––2211)) aanndd
hhyyddrraatteedd ggeerraannyyllpphheennoollss ddeerriivvaattiivveess ((2222––2266)) tthhaatt hhaavvee bbeeeenn ssttuuddiieedd iinn tthhiiss wwoorrkk..
2. Results and Discussion
2. ResultsandDiscussion
22..11.. SSyynntthheessiiss
LLiinneeaarr ggeerraannyyllaatteedd pphheennoollss//mmeetthhooxxyypphheennoolsls hhaavvee bbeeeenn ssyynntthheessiizzeedd bbyy ddiirreecctt ccoouupplliinngg ooff
ggeerraanniiooll wwiitthh ththee rersepspecetcitvive ephpehneonlo olro mr emtheothxyopxyhpenhoelnso. lTs.his Trheiasctrioenac htiaosn bheaens sbteuedniesdt ufdorie mdafnoyr
mauatnhyorsa,u tbheocrasu, sbee ciatu sies itdiirsecdtliyre ctrleylatreedla tetdo totheth esysnytnhtehseiss isoof f bbiioollooggiiccaallllyy--aaccttiivvee pphheennoolliicc
tteerrppeennooiiddss [[1111,,1144,,2200,,2266––3300]].. TThhee ccoouupplliinngg iiss ccoommmmoonnllyy ccaarrrriieedd oouutt iinn ssttrroonngg mmiinneerraall aacciiddss oorr aapprroottiicc
ssoollvveennttss wwiitthh LLeewwiiss aacciiddss,, ee..gg..,, BBFF33¨·EEtt22OO,, iinn ddiiooxxaannee ffoorr tthhee ssyynntthheessiiss ooff ttooccoopphheerroollss aanndd ggeerraannyyll aanndd
ffaarrnneessyyll ananalaolgosgso fothf etuhbei quuibnioqnueins,opn-etos,l uepn-teosluulefnonesicualfcoindiicn CacHid2 CiIn2 foCrHth2CeIs2y nftohre stihseo fscyannnthaebsiigse roofl
acanndnraeblaigteedroml aanridh urealnaatecdo nmsatirtiuheunatnsa[ 3c9o]n.sAtilttuerennatsti v[3e9ly]., ABFlt3e¨rEnta2tOiv/eAlyg, NBFO33·Eht2aOs/bAegenNuOs3e hdaass baeecnat ualsyesdt
aans da caacetatolynsitt rainleda ascaetsoonlviternilte[ a2s9 ,a3 0so].lvInentht i[s29w,3o0r]k. ,ICn othmisp wouonrdk,s C1,o3m,p14o,u1n5d,s1 71,a 3n, d141,9 1w5, e1r7e asnyndt 1h9e swizeerde
tshyrnotuhgeshiztehdis trheraocutigohn ,thuissi nregadctiiooxna,n uesainsgt hdeiosxoalvneen at,s BthFe3¨ sEotl2vOenats, tBhFe3·cEatt2aOly asst tahned ciantatlhyestp arensde nince thoer
apbresesennceceo ofra anbistreongceen oaf tam noistrpohgeerne .atmosphere.
DDiirreecctt ccoouupplliinngg bbeettwweeeenno o--ccrreessoolla annddp p-c-rcerseoslowl witihthg egrearnainoiloul nudnedrear na intriotrgoegneant matomsposhpehreerleea ldesadtos
Ctoo Cmopmoupnoudnsd14s 1a4n dan1d5 1w5i twhi3th.1 3%.1a%n dan1d2 %12y%ie yldiesl,drse,s rpeescptievcetilvye(lSyc (hSecmheem1)e. 1).
Int.J.Mol.Sci.2016,17, 840 4of18
IInntt.. JJ.. MMooll.. SSccii.. 22001166,, 1177,, 884400 44 ooff 1188

SSSccchhheeemmmeee 111... SSSyyynnnttthhheeesssiiisss ooofff CCCooommmpppooouuunnndddsss 111444,,, 111555 aaannnddd 111666...
FFoolllloowwiinngg tthhee ssaammee ssyynntthheettiicc pprroocceedduurree,, CCoommppoouunndd 1177 iiss oobbttaaiinneedd aass aa uunniiqquuee pprroodduucctt bbyy
Following the same synthetic procedure, Compound 17 is obtained as a unique product by
ccoouupplliinngg bbeettwweeeenn 22--mmeettooxxyyhhyyddrrooqquuiinnoonnee aanndd ggeerraanniiooll wwiitthh 55..99%% yyiieelldd ((SScchheemmee 22))..
couplingbetween2-metoxyhydroquinoneandgeraniolwith5.9%yield(Scheme2).

SSSccchhheeemmmeee 222... SSSyyynnnttthhheeesssiiisss ooofff CCCooommmpppooouuunnndddsss 111777 aaannnddd 111888...
SSStttaaannndddaaarrrddd aaaccceeetttyyylllaaatttiiiooonnn ( ((AAAccc222OOO///CCCHHH222CCClll222//D/DDMMMAAAPPP)) )oooff f115155 aaannnddd 111777 gggiivviveeesss tththheee aaaccceeetttyyylllaaattteeeddd dddeeerrriiivvvaaatttiiivvveeesss 111666 aaannnddd
111888 wwwiiittthhh 999444...888%%% aaannnddd 999888%%% yyyiiieeellldddsss,,, rrreeessspppeeeccctttiiivvveeelllyyy...
IIInnn ttthhheee ssseeeaaarrrccchhh fffooorrr aaa sssyyynnnttthhheeetttiiiccc pppaaattthhhwwwaaayyy tttooo ooobbbtttaaaiiinnn hhhyyydddrrraaattteeeddd gggeeerrraaannnyyylllppphhheeennnooolllsss,,, wwweee hhhaaavvveee aaatttttteeemmmpppttteeeddd
tttwwwooo dddiiiffffffeeerrreeennnttt aaapppppprrroooaaaccchhheeesss... IIInnn ttthhheee ffifiirrrsssttt ooonnneee,,, wwweee eeexxxppplllooorrreee ttthhheee pppooossssssiiibbbiiillliiitttyyy ooofff ooobbbtttaaaiiinnniiinnnggg bbbooottthhh gggeeerrraaannnyyylllaaattteeeddd
qqquuuiiinnnooonnneeesss aaannndddh yhhdyyrddarrtaaettdeeddg e grgaeenrrayanlnpyyhllppenhhoeelnnsooillnss aiinno n aae -pooonntee-s-ppyonottt h sseyysninstt.hhIeetsshiisas..s IbItte ehhnaasrse pbbeoeeretnne drreethppaootrrtsteeoddm etthhhaaytt d ssrooamtmedee
hghyeyrddarrnaaytteeloddr cggienerroaalnsnyyhllaoovrrecciibnneooellsns hhoaabvvtaeei nbbeeedeenna soombbttiaaniinonreeddp raaossd mmuciitnnsooirrn ppthrrooeddcuuocucttpss l iiinnn g tthhreeea cccootiuuoppnllioinnfggo rrrceeiaanccottiliooannn dooffg ooerrrcaciinnniooolll
aiannnddth ggeeeprraarnensiiooelnl iicnne ttohhfee 1 pp%rreeasseqennucceeeo ouoffs 11o%%xa aalqiqcuuaeecooiuudss a ootxx8aa0lliicc˝ Caacc[iid4d0 aa]t.t 88T00h °°eCCre [[f44o00r]e].., TTinhheetrrheeiffsoorareep,, p iinrno ttahhciihss ,aatpphpperrocooaaucchhp,,l ittnhhgee
crceooauucpptlilioinnngg i rsreecaaaccrttiriooiennd iisso uccaatrrurriineeddde ooruuatti ruunundsdieenrrg aaidirri ouuxssaiinnngge ddaiisootxxhaaennees oaalssv tethhneet , ssBooFllvv3ee¨nEnttt,,2 BOBFFa33··sEEttt2h2OOe aacsas tttahhleye scctaaattaanlldyyssstt m aannadldl
sasmmmaaollull naamtmsooouufnnatdtssd ooeffd aawddddaeeteddr .wwInaattteeerrr.e. sIIntnittneegrreleyss,ttitinnhggellyoyb,, ttthahieen oeobdbttpaaiirnnoeedddu pcptrrsoodddeuupccettssn dddeeoppneennthdde oocnnh etthhmeei cccahhleenmmaiitccuaarlle nnoaafttuuthrreee
oroeff a cttthhinee g rpreehaaeccnttiionnlgg. Tphphuhese,nncoooll.u. pTTlihhnuugss,b, ectcwoouueppenlliinnggge rabbneeittowwleaeeennnd 1gg,4ee-rrhaaynndiioorllo qaaunninddo n11e,,44l--ehhayydddsrrtoooqqmuuiionnnooonngee e rllaeenaayddlssa tettdoo
mhmyoodnnroooggqeeurriaannnoyynllaeatt1eedda nhhdyyddqrruooiqnquuoininneoo2nn,eea 11s waanneddll qqauusiidnnioognneeer a 22n,, yaalssa twwedeellqll uaasisn ddoiinggeeerr1aa9nn(yySllcaahtteeeddm qqeuu3ii)nn;oownnhee e 11r99e a ((sSS,ccchhoeeummpeel i n33)g);;
wbwehhtweerreeeeaanss,, 2 cc-oomuuepptlloiinxngyg h bbyeedttwwroeeqeeunni n22o--mmneeetatoonxxdyyhghyeyrddarrnooiqqouuliginniovonensee taahnneddd ggiseeurrabansntiiiootlul gtgeiivdveeqssu ttihhneeo dndieissu2u0bbssattsiitttuuhtteeeddo n qqluuyiinpnoroonndeeu 22c00t
a(aSssc ththheeem ooenn4ll)yy. Fppirrnooaddlluuy,cctht y((SSdccrhhaeteemmdeeg e44r))a.. nFFyiinnlpaahlllleyyn,, ohhlysydd22rraaattneeddd 2gg3eerwraaennryyellppohbhteeannionolelssd 22i2n2 taahnneddc o22u33 p wwlineergreer eooabbctttaaiioinnneebdde itinwn etthehnee
coco-ocuurpepsllioinnlgga nrreedaacgctteiirooannn biboeelttww(Seecehenne m oo--eccrr5ee)ss.ooll aanndd ggeerraanniiooll ((SScchheemmee 55))..
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Scheme 3. Synthesis of Compounds 1, 3 and 19.
In this reaction, Compounds 1, 3 and 19 were obtained with 28.0%, 7.6% and 1.9% yields,
respectively. When this coupling is carried out in the presence of a nitrogen atmosphere and with no
IInntt.. JJ.. MMooll.. SSccii.. 22001166,, 1177,, 884400 55 ooff 1188
added water, Compound 1 is obtained as the exclusive product [26]. Recently, this reaction has been
performed at higher temperatures using aluminum phenoxide as the catalyst, and a completely
Scheme 3. Synthesis of Compounds 1, 3 and 19.
different pattern of products has been reported. Compound 1 and a mixture of digeranylated
quinones (19 and di-ortho-geranylated quinone) were obtained with 40% and 27% yields,
In this reaction, Compounds 1, 3 and 19 were obtained with 28.0%, 7.6% and 1.9% yields,
respectively, but Compound 3 was not identified [41].
respectively. When this coupling is carried out in the presence of a nitrogen atmosphere and with no
On the other hand, a methoxy substitution in the hydroquinone induces a complete change in
added water, Compound 1 is obtained as the exclusive product [26]. Recently, this reaction has been
the product distribution, i.e., Compound 20 is obtained with 4.1% yield.
performed at higher temperatures using aluminum phenoxide as the catalyst, and a completely
different pattern of products has been reported. Compound 1 and a mixture of digeranylated
quinones (19 and di-ortho-geranylated quinone) were obtained with 40% and 27% yields,
respectively, but Compound 3 was not identified [41].

On the other hand, a methoxy substitution in the hydroquinone induces a complete change in
the product distribution, i.e.,S Cchoemmpeo3u.nSdyn 2th0e issi soobftaCionmedp owunitdhs 41.,13%a nydie1l9d..
Scheme 3. Synthesis of Compounds 1, 3 and 19.

In this reaction, CompoundSsc h1e,m 3e a4.n Sdy n1t9h ewsise roef Coobmtapionuendd w20i.t h 28.0%, 7.6% and 1.9% yields,
respectively. When this coupling is carried out in the presence of a nitrogen atmosphere and with no
addeIdt iws awteorr,t hC momenptoiounnidn g1 tihs aotb gtearinaendy laaste tdh eq uexincolunseisv e(3 p, 1ro9d, 2u0c)t a[r2e6 ]f.o Rrmeceedn otlnyl,y t hbiys croeaucptliionng h gaesr abneieonl
wpeirthfo 1r,m4-eddi haytd hroigxhyebre ntezmenpee sryastutermess .u Psrionbga ablluym, thineu omxi dpahteionno xtoid 1e,4 a-qs utihneo ncea tiasl eynsth, aanncded a b yco tmhep rleedteolyx
pdrifofpereerntite sp oaft theyrnd roofq upirnoodnuec ctso mhpaso ubnedesn. reported. Compound 1 and a mixture of digeranylated
quinoFnineasl ly(1, 9in tahned c oduip-olirnthgo o-fg gerearnanyiloalt ewdi thq uo-icnroenseo)l , hwyedrrea teodb tCaionmedp ouwnitdhs 2420 a%n da 2n3d w e2r7e% o bytaieinldesd,
Scheme 4. Synthesis of Compound 20.
rweistphe 9c.t0iv%e layn, db u1t0 C.5o%m ypieoludnsd, r 3e swpSeaccsht nievometl eyid4. .eSnytinftiheeds i[s4o1f].C ompound20.
OIt nis twheo rotthh mere hnatinodn,i nag m theatht ogxeyra snuyblastteitdu tqiuonin ionn tehse ( 3h, y1d9,r 2o0q)u ainreo fnoer mineddu ocensl ya b cyo cmopulpeltien gch gaenrgane iionl
twhiet hp r1o,4d-udciht yddisrtorixbyubteionnz,e ni.ee. ,s Cysotmempso.u Pnrdo b2a0b ilsy o, tbhtea ionxeidd awtiiothn 4to.1 1%,4 y-qieulidn.o ne is enhanced by the redox
properties of hydroquinone compounds.
Finally, in the coupling of geraniol with o-cresol, hydrated Compounds 22 and 23 were obtained
with 9.0% and 10.5% yields, respectively.

Scheme 5. Synthesis of Compounds 22 and 23 by coupling of o-cresol and geraniol, in the presence of
Scheme5.SynthesisofCompouSncdhsem22ea 4n. dSy2n3tbhyescios uopf lCinogmopfoou-cnrdes 2o0l.a ndgeraniol,inthepresenceof
water and the absence of a nitrogen atmosphere.
waterandtheabsenceofanitrogenatmosphere.
It is worth mentioning that geranylated quinones (3, 19, 20) are formed only by coupling geraniol
The formation of Compounds 22 and 23 may be explained by the proposed mechanism depicted
withI 1n,4t-hdiishyredarcotxioynb,enCzoemnep soyusntedmss1. ,P3roabnadbly1,9 thwee orxeidoabttiaoinn etod 1w,4i-tqhui2n8o.0n%e i,s 7e.n6h%anacnedd b1.y9 %they rieedldosx,
in Scheme 6.
rpersoppeecrttiiveesl yo.f Whyhdernoqtuhiisnocnoeu pcolimngpoisucnadrsr.i edoutinthepresenceofanitrogenatmosphereandwith
noadFSdicnehdaelmlwye,a 5itn.e S rty,hnCet ohcmeosuipsp ooluifn nCgdo om1fp gioseuronabdntsia o2iln2 w eadnitdha s2 o3t- hcbryee cseooxulc,pl huliysnidgvr oeaftp oer-dco rdCesuoocmlt ap[n2od6u ]gn.edrRasen 2cio2eln ,a tinnlyd t, h2teh3 pi wsreresereaenc coteib ootnaf inheads
bweietnh wp9.ea0rt%efor raamnndde dt1h0ae.t 5ah%bisg eyhniceeerld otsef ,ma r nepsietprroeagctuetinrv eeasltymu.s oisnpghearleu. minumphenoxideasthecatalyst,andacompletely
differentpatternofproductshasbeenreported. Compound1andamixtureofdigeranylatedquinones
(19 anTdhed fio-ormrthaoti-ogne roaf nCyolamtepdouqnudins o2n2 ea)nwd e2r3e moabyta bine eedxpwlaiitnhe4d0 b%y athned p2r7o%poyseiedl dmse,crheaspneiscmtiv deelpy,icbtuedt
Cino mScphoeumned 63. wasnotidentified[41].
Ontheotherhand,amethoxysubstitutioninthehydroquinoneinducesacompletechangeinthe

productdistribution,i.e.,Compound20isobtainedwith4.1%yield.
Itisworthmentioningthatgeranylatedquinones(3,19,20)areformedonlybycouplinggeraniol
with1,4-dihydroxybenzenesystems. Probably,theoxidationto1,4-quinoneisenhancedbytheredox

propertiesofhydroquinonecompounds.
FSicnhaelmlye, i5n. Sthynetchoesuips loifn Cgoomfpgoeuranndiso 2l2w anitdh 2o3- cbrye scooul,phliyndgr oaft eod-crCesooml apnodu gnedrasn2i2ola, innd th2e3 pwreesreenoceb otaf ined

withw9.0at%er aanndd t1h0e. 5a%bseyniecled osf, ar ensiptreocgteinv ealtym. osphere.
TheformationofCompounds22and23maybeexplainedbytheproposedmechanismdepicted
inSchTehme efo6r.mation of Compounds 22 and 23 may be explained by the proposed mechanism depicted
in ScIhnemthee 6fi.r ststep,anallyliccarbocationisformedbythereactionofBF ¨Et Owithgeraniol,which
3 2
isthencoupledwithphenolviaElectrophilicAromaticSubstitution(EArS)(Step2). Inpresenceof
water,theadductBF ¨H Oispresumablyformedbynucleophilicdisplacementofanethermoleculeby
3 2
H O(Step3). Subsequently,thisadductreactswiththegeranylchainbyaMarkovnikov-typeaddition,
2
forming a stable tertiary carbocation, which is then hydrated by reaction with a water molecule
Int.J.Mol.Sci.2016,17, 840 6of18
(Steps 3 and 4). Finally, the remaining olefinic bond is hydrated by BF ¨H O, and a completely
3 2
hydratedgeranylchainisobtained(Step5). Itisworthmentioningthatwaterisadded24hafterthe
couplingreactionhasbeenstarted. ThismeansthatStep3beginswhenmostofthegeranylphenolhas
aInltr.e Ja. Mdyol.b Secie. n20f1o6r, m17,e 8d4.0 6 of 18

SScchheemmee 66.. PPrrooppoosseedd mmeecchhaanniissmm ffoorr tthhee ffoorrmmaattiioonn ooff CCoommppoouunnddss 2222 aanndd 2233..
BIna sthede foinrstth sitsepre, saunlt a,lolyulrics eccaornbdocaaptiporno aisc hfocromnesdis tbsyo tfhteh eredaicrteicotnh oyfd BraFt3i·oEnt2Oof wthieths igdeeracnhiaoinl, wbyhtihche
rise atchteionn coofuaplgeedr awniytlha tpedhepnhoeln voilaw EiltehctaroLpehwiilsica Acirdo(mBFati¨cE StuOb)stiintudtiiooxna (nEeAarnSd) i(nSttehpe 2p)r.e Isne npcreesoefnwcea toerf.
3 2
Cwoamterp,o tuhne dad1d7uwcta sBFsu3·bHm2Oit tiesd prtoestuhmisarbelayc tfioornm,eadn dbyc onmucploeuopnhdisli2c1 d,i2s4p–l2a6cewmeerneto obft aanin eetdhewr imtho1le1c.1u%le,
1b0y. 7H%2,O2 4(%Steapn d3)1. 8S.9u%bsyeiqeuldens,tlrye,s pthecist ivadeldyu(cStc hreeamctes 7w).ith the geranyl chain by a Markovnikov-type
additCioonm, pfoorumndinsg2 4a asntdab2l6e mtearytiabreyf ocramrbeodcathtiroonu,g whhStiecphs i3s –t5hoefnt hheymdreactheadn bisym rpearocptioosne dwiinthS cah ewmaete6r.
Hmoowleecvueler, i(nSttehpiss r3e aactniodn ,4C). oFminpaolulyn,d t2h5e isrefmoramineidngb yoclyecfilniziac tiboonnodf tihse hteyrdtiraartyedc abrbyo cBaFti3o·Hn2fOor, maendd ina
Scotempp3l,ettheelyf ohrymdartaitoendo gfetrearntiyarl ychcaarinb oisc aotbiotanininedth (eSCte-p71 5p).o Isti tiiso wnoorftghe mraennytlicohnaining, 6th-eant dwoa-ttreirg isc yacdlidzeadti o2n4
fhr oamftetrh ethCe2 c1-oCu3p1ldinogu breleacbtoionnd haansd bheyednr asttiaorntebdy. tThheiss umbeseaqnus etnhtant uSctelepo p3h bileigciantst awckhoenf wmaotesrt oonf tthhee
tgeerrtaianryylpcahrebnoocla htiaosn ailnretahdeyC b-e3e1np ofosirtmioend(.S cheme8).
TBhaseecda robno cthatiiso rnesinutlet,r mouerd isaetceosnadp paepaprrionagcihn cSocnhseimstess o6f athned d8ihreacvte hbyedernatpiroonp oofs etdhef osridceo ucphlaiinng boyf
pthhee nreoalcwtiiotnh ogfe raa ngieorlaannydlavteadr ipouhesnreoal cwtiiotnhs ao Lfegweriasn aicoildi n(BaFc3i·dEitc2Oaq) uineo duisoxsoanluet iaonnd[ 3in9 ,4th0e]. presence of
wateCr. oCmopmopuonudnsd1 147– 1w8,a2s 0s,u2b2m–2it6teadr eton ethwi,s arneadctthioenir, asntrdu cctoumraplocuhnadrasc 2te1r, i2z4a–ti2o6n wiserdee oscbrtiabinededi nwtihthe
n11e.x1t%se, c1t0i.o7n%., 24% and 18.9% yields, respectively (Scheme 7).
Int. J. Mol. Sci. 2016, 17, 840 7 of 18
Int.J.Mol.Sci.2016,17, 840 7of18
Int. J. Mol. Sci. 2016, 17, 840 7 of 18

Scheme 7. Synthesis of Compounds 21 and 24–26.
Compounds 24 and 26 may be formed through Steps 3–5 of the mechanism proposed in
Scheme 6. However, in this reaction, Compound 25 is formed by cyclization of the tertiary

carbocation formed in Step 3, the formation of tertiary carbocation in the C-7′ position of geranyl
Scheme 7. Synthesis of Compounds 21 and 24–26.
chain, 6-endo-trig cyclizatiSocnh efmroem7. Sthyne thCe2si′s-Co3f′C odmoupboulen dbso2n1da nadn2d4 –h26y.dration by the subsequent
nucleophilic attack of water on the tertiary carbocation in the C-3′ position (Scheme 8)
Compounds 24 and 26 may be formed through Steps 3–5 of the mechanism proposed in
Scheme 6. However, in this reaction, Compound 25 is formed by cyclization of the tertiary
carbocation formed in Step 3, the formation of tertiary carbocation in the C-7′ position of geranyl
chain, 6-endo-trig cyclization from the C2′-C3′ double bond and hydration by the subsequent
nucleophilic attack of water on the tertiary carbocation in the C-3′ position (Scheme 8)

SScchheemmee 88.. PPrrooppoosseedd mmeecchhaanniissmm ffoorr ffoorrmmaattiioonn ooff CCoommppoouunndd 2255.. RReeaaccttiioonn ooffL Leewwisisa accidid( B(BFF¨3·EEtt2OO))
3 2
wwiitthh wwaatteerr,, aaddddiittiioonn MMaarrkkoovvnniikkoovv ooff HH bbyy BBFF3HH2OO aadddduucctt oonn CC--661′ ppoossiittiioonn ooff ggeerraannyyll cchhaaiinn aanndd
3 2
ccaarrbbooccaattiioonn ffoorrmmaattiioonn iinn CC--771′,, 66--eennddoo--ttrriigg ccyycclliizzaattiioonn aanndd llaatteerr ggeerraannyyll cchhaaiinn hhyyddrraattiioonn..

The carbocation intermediates appearing in Schemes 6 and 8 have been proposed for coupling
2.2. StructureDetermination
of phSenchoel mwei t8h. gPerorpanosioedl amnedc hvaanriisomu sf orre afocrtmioantsio onf ogfe Craonmioplo iunn dac 2id5.i cR aeaqcutieoonu osf s Loeluwtiiso anc i[d3 9(,B4F03]·.E t2O)
TCwhoietmhc phwoeamutenircd, asal d1sdt4ri–uti1oc8tnu, 2rMe0sa, ro2kf2oa–vl2nl6ick oaomrve p onof euHwn d,b ysa nsBydFn 3tHthh2eeOisr i azsdetrdduuicncttu tohrnais lC wc-h6oa′r rpkaowcstieetirroeinzm aotafi oignnel yriasen sdytlea sbcchlriasiibhnee dadn bidny t1hDe
and2cDarNbouccalteioanr fMoramganteiotnic inR eCs-o7n′, a6n-ecned(oN-tMrigR c)yscplizeacttiroons caonpd ylatteecr hgneriaqnuyels c.hAailnl NhyMdrRatsiopne.c traaregivenin
next section.
FigureS1. Inthissection,theNMRdatausedtodeterminethechemicalstructureofnewcompounds,
g2e.2r.a SnTtyrhluaect tecudarrepb hDoeecntaeotrilmodnien riainvttiaoetnrim veesd(i1a4t–e1s 8a)p,gpeeraarninygla itned Sqchueinmoense 6(2 a0n)dan 8d hhayvder abteeedng perroanpyolspehde fnoorl sc(o2u2p–l2i6n)g,
aorfe pdhisecnuosls ewditihn gdeertaainl.ioCl oamndp ovuanridou19s hreaascbtieoennsa olrfe gaedryanreioplo irnte adc,ibduict aitqhuaesobuese snoilnuctliuodne [d39in,4t0h]i. ssection
becauTCshoeemt hcpheoeNumnMicdaRsl 1as4strs–ui1gc8nt,u a2rt0eio,s n 2o2fg– ia2vl6le ncaorimen nplieotweurn,a datuns rdsey tinhstenhioer stsirtzriegudhc ttiun[r4 ta1hl] i.csCh waoromarckpt eowruieznradet simo1na4 i–ins1l 8yd: eeTsschtareibb1leiHsdh- NeindM tbhRye
s1npDeex cattn rsudem c2tDioo fnNC. uocmlepaor uMnadg1n4etsihco Rwessoanpaantcteer (nNcMhaRra) cstpeercisttriocsocfoapryo mtecahtincitqrui-esus.b Astlilt uNtMionR, si.pe.e,cstirnag alerte sgigivneanl
in Figure S1. In this section, the NMR data used to determine the chemical structure of new
at6.93ppm(1H,H-3, meta-couplingofH-3withH-5wasnotdetected); doubletat6.88ppm(1H,
c2o.2m. pStoruunctdusr,e Dgeertaernmyilnaatetido n phenol derivatives (14–18), geranylated quinone (20) and hydrated
J = 8.0, H-5); and doublet at 6.69 ppm (1H, J = 8.0, H-6). The position of the geranyl chain on the
geranylphenols (22–26), are discussed in detail. Compound 19 has been already reported, but it has
aromaTthice rcinhgemhaicsabl esetrnuecsttuarbelsi sohfe adllb cyotmwpoo-duinmdesn ssyionnthales(2izDe)dH ine ttehriosn wucolreka rwMeruel tmipaleinBlyo nedstCabolrirsehleadti obny
(b1HeDeM naB niCdn )c2lcDuod rNeredul cailtneioa trnh sMi.saI nsgentchettiiisocns Rp beesecoctranuuamnsec, eat h(2NeJ HMN-CRMc) oRsup aepsclsitnrigognscoaoftipHoyn- 1t eg1cwihvnietnihq uCine- s4l.i (AtδeClrla =NtuM1r3eR3 .i 9ssp pnepocmttr ar)i aagrnhedt g[Ci4v-1e2]n1.
(CiδnoC mF=ipg1ou2ur3ne.6 dSsp1 p.1 m4I–n)1 a8tnh: idTs hasee3 cJ1HtHi-oC-nNc, oMtuhRpe lisNnpgMecbRtre utdwmaet eaon fu tCsheeodms itpgoon uadlnesdteo rf1m4C i-ns1eh, oCtwh-3es, cCah -ep3m1ataitcneadrln C sct-rh5uaacrttaucδrtCee r=iosf1t i3cn0 e.o9wf,
aromatic tri-substitution, i.e., singlet signal at 6.93 ppm (1H, H-3, meta-coupling of H-3 with H-5 was
1c3o5m.7paonudnd12s6, .7geprpamny,lraetsepde cptihveenlyo,lw deererivoabtsievrevse d(.14F–u1r8th),e rgceorarrneylalatitoedns qatu3inJHo-nCeb e(2tw0)e eanntdh ehCyHdr3a-tAedr
not detected); doublet at 6.88 ppm (1H, J = 8.0, H-5); and doublet at 6.69 ppm (1H, J = 8.0, H-6). The
ggreoruapny(lδpHhe=n2o.l2s3 (2p2p–m26))w, airteh dCi-s1cuasnsdedC i-n3 daettδaCil.= C1o5m1.p8oaunndd1 1390 .h9aps pbmee,nr easlpreeacdtiyv erelyp,owrteerde, obbuste irtv heads
position of the geranyl chain on the aromatic ring has been established by two-dimensional (2D)
(bFeigeun rein3calu).deOdn itnh ethoitsh eserchtiaonnd ,btehceau1sHe -NthMe RNMspRec tarsusmignoaftiCoonm gpivoeunn din1 5litsehraotwusrea issi nngolte trisgighnt a[l41a]t.
6HC.9eo3tmeprpoponmuunc(dl1esHa r1, M4H–u-138l,t:i mpTleeht eBa -o1cHnodu-N pClMoinrRgre losapftieHocnt-r3 u(HwmMi thoBfC HC)- 5ocomwrrpaeoslauntnioodtn ds1.e4 It nesc htthoeiwds) ss apane dcptrtauwttmeor, nda o 2cuJhHba-Clre actocstuiegprnilsiantligcs aootff

6a.r9o2mpaptmic t(r1i-Hsu,bJs=tit8u.7tioHnz, ,i.He.,- 5si)nagnledt 6si.7g3na(l1 Hat ,6J.9=3 8p.p7mH (z1,HH, -H6)-.3,A mdedtiat-icoonuaplllyi,nag osifg Hna-3l awpipthe aHr-in5 gwaats
5n.0o7t pdpetmec(tse,d1)H; d)wouabslaests aigt n6e.8d8t oppthme O(1HH,g Jr o=u 8p..0,T Hhe-5a)r;o amnadt idcosuubblsetitt uatt i6o.n69p aptptemrn (s1hHo,w J s=u 8n.0e,q Hui-v6o).c aTlhlye
position of the geranyl chain on the aromatic ring has been established by two-dimensional (2D)
Heteronuclear Multiple Bond Correlation (HMBC) correlations. In this spectrum, a 2JH-C coupling of
IInntt.. JJ.. MMooll.. SSccii.. 22001166,, 1177,, 884400 88 ooff 1188
HH--11′′ wwiitthh CC--44 ((δδCC == 113333..99 ppppmm)) aanndd CC--22′′ ((δδCC == 112233..66 ppppmm)) aanndd aa 33JJHH--CC ccoouupplliinngg bbeettwweeeenn tthhee ssiiggnnaallss ooff
CC--11,, CC--33,, CC--33′′ aanndd CC--55 aatt δδCC == 113300..99,, 113355..77 aanndd 112266..77 ppppmm,, rreessppeeccttiivveellyy,, wweerree oobbsseerrvveedd.. FFuurrtthheerr
ccoorrrreellaattiioonnss aatt 33JJHH--CC bbeettwweeeenn tthhee CCHH33--AArr ggrroouupp ((δδHH == 22..2233 ppppmm)) wwiitthh CC--11 aanndd CC--33 aatt δδCC == 115511..88 aanndd
113300..99 ppppmm,, rreessppeeccttiivveellyy,, wweerree oobbsseerrvveedd ((FFiigguurree 33aa)).. OOnn tthhee ootthheerr hhaanndd,, tthhee 11HH--NNMMRR ssppeeccttrruumm ooff
CCoommppoouunndd 1155 sshhoowwss aa ssiinngglleett ssiiggnnaall aatt 66..9933 ppppmm ((11HH,, HH--33,, mmeettaa--ccoouupplliinngg ooff HH--33 wwiitthh HH--55 wwaass nnoott
ddeetteecctteedd)) aanndd ttwwoo ddoouubblleett ssiiggnnaallss aatt 66..9922 ppppmm ((11HH,, JJ == 88..77 HHzz,, HH--55)) aanndd 66..7733 ((11HH,, JJ == 88..77 HHzz,, HH--66))..
Int.J.Mol.Sci.2016,17, 840 8of18
AAddddiittiioonnaallllyy,, aa ssiiggnnaall aappppeeaarriinngg aatt 55..0077 ppppmm ((ss,, 11HH)) wwaass aassssiiggnneedd ttoo tthhee OOHH ggrroouupp.. TThhee aarroommaattiicc
ssuubbssttiittuuttiioonn ppaatttteerrnn sshhoowwss uunneeqquuiivvooccaallllyy tthhaatt tthhee ggeerraannyyll cchhaaiinn iiss aattttaacchheedd ttoo tthhee oorrtthhoo ppoossiittiioonn aatt
thhhyyaddtrrtoohxxeyygll e ggrrraoonuuypplscs..h TTaihhnee i sppooatsstiiattiicoohnne doofft otthhtehe eggoeerrratahnnoyyllp ccohhsaiatiiinno noonant tthhhyeed aarrroooxmmylaagttiircco ruriipnnsgg. hThaahsse bbpeeoeesnnit iccooonnnoffiifrrmtmheeeddg ebbryya n22yDDl
cHHhMMaiBnBCCo nccoothrrrreeeallaartotiimoonnasst..i cIInnr i tnthhgiissh ssappseebccetterruunmmco,, ntthhfieer m ssiieggdnnaablly aa2tt D δδHHH ==M 33B..33C55 cppopprmmre laaasstissoiiggnnns.eedIdn tttooh HiHs--s11p′′ e((c22tHHru ddm,, ,JJ t ==h e77..s22i g HHnzaz)l)
asshhtooδwHwss= 33J3JHH.3--CC5 ccpoopuupmplliiannsggs iwwgniittehhd CCt--o11 H((δδ-CC1 ==1 (11255H22..11d)),,, JCC=--337 ((.δ2δCCH ==z 11)22s77h..88o))w aasnnd3dJ HCC---C33′′c ((oδδuCC p ==l i11n33g88..w11 ppitpphmmC))- 1aann(δddC 22JJ=HH––1CC5 cc2oo.u1u)pp,llCiinn-g3g
(wwδCiitthh= CC12--227 . a8an)naddn CCd--C22′′- 3((δδ1CC( δ ==C 11=22661..3668 aa.1nnpddp 11m2211)..a88n ppdpp2mmJH,, –rrCeesscppoeeuccpttilivivneegllyyw;; FiFtiihgguCurr-ee2 3a3bnb)d) AACdd-2dd1ii(ttδiiooCnn=aall1llyy2,6, t.t6hheea nssdiigg1nn2aa1ll . 8aattp δδpHHm ==,
r33e..33s5p5 eppcpptimmve ((lHHy;--11F′′i))g ssuhhrooeww3ebedd) Assppdaadttiiiataillo ccnooarrlrrleeyll,aattthiiooennsssig wwniiattlhha ttthhδeeH ssii=ggnn3aa.3lls5s aapttp δδmHH ==( H 66..-991331),, 55s.h.00o77w aanenddd s11p..77a99t ippaplpcmmo,,r araesslssaiigtginnoeendds
wttooi tHHh--t33h,,e OOsHHig n aaannldsd aCCtHHδH33--CC=336′′.,,9 rr3ee,ss5pp.e0ec7cttaiivvneedllyy1;;. 7ww9hhpiiplleem tthh,aees sssiiigggnnnaaeldl aattto δδHHH -==3 ,22O..22H33 ppanppdmmC ((Hss,, 3CC-CHH3331--,AArerr)s) pssehhcootwwiveeeddly s;sppwaahttiiialaell
tcchooerrrrseeillgaanttiiaoolnnasst wwδHiitthh= HH2.-2-333 aapnnpddm HH(--s55, C((FFHiigg3uu-Arreer )33scch))..o wedspatialcorrelationswithH-3andH-5(Figure3c).

FFFiiiggguuurrreee 333... MMMaaaiiinnn ooobbbssseeerrrvvveeeddd cccooorrrrrreeelllaaatttiiiooonnnsss::: 222DDD HHHeeettteeerrrooonnnuuuccclleleeaaarrr MMMuuullltttiiipppllelee BBBooonnnddd CCCooorrrrrreeelllaaatttiiiooonnn (((HHHMMMBBBCCC))),,,
CCCooommmpppooouuunnnddd 111444 (((aaa))) aaannnddd CCCooommmpppooouuunnnddd 111555 (((bbb)));;; 111DDD NNNuuucccllleeeaaarrr OOOvvveeerrrhhhaaauuussseeerrr EEEffffffeeecccttt SSSpppeeeccctttrrrooossscccooopppyyy (((NNNOOOEEESSSYYY)))
CCCooommmpppooouuunnnddd 111555 (((ccc)))...
IIInnn ttthhheee 111HHH---NNNMMMRRR ssspppeeeccctttrrruuummm ooofff ttthhheee aaaccceeetttyyylllaaattteeeddd dddeeerrriiivvvaaatttiiivvveee 111666,,, aaa sssiiinnngggllleeettt a aattt δ δδHH === 222...222999 ppppppmmm (((333HHH,,, CCCHHH33CCCOOO)))
H 3
wwwaaasss ooobbbssseeerrrvvveeeddd... AAAddddddiiitttiiiooonnnaaallllllyyy,,, i iinnnt tthhheee1 11333CCC NNNMMMRRR ssspppeeeccctttrrruuummm,,, ttthhheee sssiiigggnnnaaalllsss a aappppppeeeaaarrriininnggga aatttδ δδCC === 111666999...666 (((CCC===OOO))) aaannnddd
C
222000...888 (((CCCHHH33))) ppppppmmm cccooonnnfffiiirrrmmmeeeddd ttthhheee ppprrreeessseeennnccceee ooofff mmmooonnnoooaaaccceeetttyyylllaaattteeeddd dddeeerrriiivvvaaatttiiivvveee 111666...
3
CCCooommmpppooouuunnnddd 111777::: IIInnn ttthhheee 111HHH---NNNMMMRRR ssspppeeeccctttrrruuummm,,, aaa pppaaatttttteeerrrnnn ccchhhaaarrraaacccttteeerrriiissstttiiiccc ooofff aaarrrooommmaaatttiiiccc ttteeetttrrraaa---sssuuubbbssstttiiitttuuutttiiiooonnn,,,
ii..ee..,, ttwwoo ssiinngglleett ssiiggnnaallss aatt 66..6677 ((11HH,, HH--33)) aanndd 66..4433 ((11HH,, HH--66)),, wwaass oobbsseerrvveedd.. TThhee ppoossiittiioonn ooff tthhee ggeerraannyyll
i.e.,twosingletsignalsat6.67(1H,H-3)and6.43(1H,H-6),wasobserved. Thepositionofthegeranyl
cchhaaiinn oonn tthhee aarroommaattiicc rriinngg wwaass eessttaabblliisshheedd bbyy ttwwoo--ddiimmeennssiioonnaall ((22DD)) HHMMBBCC ccoorrrreellaattiioonnss.. IInn tthhiiss
chain on the aromatic ring was established by two-dimensional (2D) HMBC correlations. In this
ssspppeeeccctttrrruuummm,,, aaa 222JJJHH––CC cccooouuupppllliiinnnggg ooofff HHH---111′′1 wwwiiittthhh CCC---222 (((δδδCC === 111111888...555 ppppppmmm))) aaannnddd CCC---2221′′ (((δδδCC === 111222111...888 ppppppmmm)))a aannnddda aa3 J33JJHH––CC
H–C C C H–C
cccooouuupppllliiinnnggg bbbeeetttwwweeeeeennn ttthhheee sssiiigggnnnaaalllsss ooofff CCC---111,,, CCC---333 aaannnddd CCC---3331′′ aaattt δδδCC === 111444777...666,,, 111111555...333 aaannnddd 111333999...222 ppppppmmm,,, rrreeessspppeeeccctttiiivvveeelllyyy,,,
C
wwweeerrreee ooobbbssseeerrrvvveeeddd... IIInnn aaaddddddiiitttiiiooonnn,,, aaa c ccooorrrrrreeellalaattitioioonnna aattt3 33JJJHH––CC bbbeeetttwwweeeeeennn ttthhheee CCCHHH33OOO gggrrrooouuuppp (((δδδHH === 333...888333 ppppppmmm))) aaannnddd CCC---555
H–C 3 H
(((δδδCC === 111444555...444 ppppppmmm))) aaannnddd cccooorrrrrreeelllaaatttiiiooonnnsss bbbeeetttwwweeeeeennn 222JJJHH––CC aannadndd 33JJ3HHJ––CC ooff otthhfeet h OOeHHO--HCC4-4C gg4rrooguuroppu ((pδδHH( δ == 55=..11555 .pp15ppmmpp)) mww)iittwhh iCCth--
C H–C H–C H
C33 -((3δδCC( δ== 11=11551..313 5 pp.3ppmmpp)) m ww)eewrreee aarellssoaol sooobbssoeebrrsvveeerddv e((dFFiig(gFuuirgreeu 44re)).. 4).
C

FFFiiiggguuurrreee 444... MMMaaajjjooorrr 222DDD HHHMMMBBBCCC ooobbbssseeerrrvvveeeddd cccooorrrrrreeelllaaatttiiiooonnnsss fffooorrr CCCooommmpppooouuunnnddd 111777...
IIInnn ttthhheee 111HHH---NNNMMMRRR ssspppeeeccctttrrruuummm ooofff ttthhheee aaaccceeetttyyylllaaattteeeddd dddeeerrriiivvvaaatttiiivvveee 111888,,, tttwwwooo sssiiinnngggllleeettt sssiiigggnnnaaalllsss aaattt δδδHHH === 222...222999 aaannnddd
222...222888 ppppppmmm (((eeeaaaccchhh 333HHH,,, CCCHHH333CCCOOO))) wwweeerrreee ooobbbssseeerrrvvveeeddd... AAAddddddiiitttiiiooonnnaaallllllyyy,,, iiinnn ttthhheee 111333CCC NNNMMMRRR ssspppeeeccctttrrruuummm,,, ttthhheee sssiiigggnnnaaalllsss
aaappppppeeeaaarrriiinnnggg aaattt δδδ === 111666999...222 (((CCCOOOCCCHHH333---CCC444))),,, 111666888...999 (((CCCOOOCCCHHH333---CCC111))) ppppppmmm aaannnddd δδδ === 222000...888 (((CCCHHH333CCCOOOOOO---CCC111))) aaannnddd
222000...666 (((CCCHHH333CCCOOOOOO---CCC444))) ppppppmmm,,, cccooonnnfffiiirrrmmmeeeddd ttthhheee ppprrreeessseeennnccceee ooofff dddiiiaaaccceeetttyyylllaaattteeeddd dddeeerrriiivvvaaatttiiivvveee 111888...
CCCooommmpppooouuunnnddd 111999::: TTThhheee sssyyymmmmmmeeetttrrriiciccaaalllm mmooolelleecccuuulallaarrrs tssrtturruuctccuttuurerreew wwasaassc occnoofinnrffmiirrmmedeeddb y bbtyyh ttehhseeu ssbuusbbtisstuttiitttiuuottniioopnna ppttaaettrttneerrinnn
tiinhn e tthhoeel e oofilleenffiiznno zznooennaeen aadnnbddy bbtyyh ttehhieen iitnnettneesnnistsyiittyyo fooiffn iintnettgeerggarrtaaettdeedds issgiiggnnnaalasllsso oofffh hhyyydddrroroogggeeennna aattotoommmsssi iinnn q qquuuiiinnnooonnneee aaannnddd ooollleeefiffiinnniiiccc
pppooorrrtttiiiooonnn... FFFooorrr iinninsssttaatannnccceee,, ,tthhthee e ssiisggingnanalla aaltta δδtHHδ H== 66=..77600. 7pp0pppmmp ((mss,, 2(2sHH, ,2, HHH-,-33H aa-nn3dda HHnd--66H)) ii-nn6dd)iiiccnaadtteeicss a ttthheees ppthrreeesspeennreccseee oonffc ettwwooof
two identical H. In a previous report, two different signals were found and assigned to these H

(δ =6.48ppm,1H,s,H-13andδ =6.52ppm,1H,s,H-16).Adetailedassignmentof1H-NMRsignals
H H
isgivenintheexperimentalpart,andthecorrespondingspectrumisshownintheSupplementary
Material. Additionally,spatialcorrelations(NOE)wereobservedforthesignalsatδ =6.70ppmand
H
atδ =3.21ppm(4H,d,J=6.8,H-11)andforthelatterandthesignalatδ =1.73ppm,assignedto
H H
Int. J. Mol. Sci. 2016, 17, 840 9 of 18
identical H. In a previous report, two different signals were found and assigned to these H (δH =
6.48 ppm, 1H, s, H-13 and δH = 6.52 ppm, 1H, s, H-16). A detailed assignment of 1H-NMR signals is
given in the experimental part, and the corresponding spectrum is shown in the Supplementary
Int.J.Mol.Sci.2016,17, 840 9of18
Material. Additionally, spatial correlations (NOE) were observed for the signals at δH = 6.70 ppm and
at δH = 3.21 ppm (4H, d, J = 6.8, H-1′) and for the latter and the signal at δH = 1.73 ppm, assigned to
CCHH33--CC33′1 ((FFiigguurree 55aa)). .FFininaalllyly, ,inin ththee 131C3C NNMMRR spsepcetcrturmum, o,nolnyl yonoen seigsinganl aalt aδtCδ =C 1=871.867 p.6ppmp (mC-(1C a-1nda nCd-
C4)- 4o)f othfeth cearcbaorbnoynl yglroguropu wpaws aosbosbersverevde, dco,cnofinrfimrimngin tghteh seysmymmemtreitcraicl asltrsutrcutuctruer oefo CfoCmompopuonudn d191. 9.

FFiigguurree 55.. MMaaiinn oobbsseerrvveedd ccoorrrreellaattiioonnss:: 11DD NNOOEE CCoommppoouunndd 1199 ((aa)),, 22DD HHMMBBCC CCoommppoouunndd 2200 ((bb))..
Compound 20: The presence of double geranyl chain substitution on the quinone nucleus was
Compound20: Thepresenceofdoublegeranylchainsubstitutiononthequinonenucleuswas
ccoonnffiirrmmeedd bbyy tthhee oobbsseerrvvaatitoionn oof ftwtwo ododuobulbelte stigsinganlsa lisn itnheth 1eH1-NHM-NRM sRpescptreucmtru amt δaHt =δ 3.1=5 p3.p1m5 p(2pHm,
H
(J2 =H 7,.J4 =H7z.)4 aHndz) 3a.1n2d p3p.1m2 p(2pHm, J( 2=H 7,.0J =Hz7).0, wHhzi)c,hw wheicrhe wasesrigenaesds igton ethdet ohytdhreohgyendsr oHge-1n′s aHnd-1 H1 a-n2′d′,
Hre-s2p1e1,crtievsepleyc. tiAvedldy.itiAondadliltyio, nthalel y,ptrheesepnrcees eonfc eonolfyo nolnye ohnyedhryodgreong eant aδtH δ= =6.362.3 2pppmpm (1(1HH, ,ss, ,HH--66))
H
demonstrates the degree of tetra-substitution on the quinone moiety. Differentiation between geranyl
demonstratesthedegreeoftetra-substitutiononthequinonemoiety. Differentiationbetweengeranyl
cchhaaiinnss wwasase staebstlaisbhleisdhebdy thbeyH MthBe CHcoMrrBeCla tiocnorsroeblasteirovnesd foobrsHer-v1e1dat 3fJor Hw-i1th′ Ca-t4 (3δJH–C= 1w87it.h9 ppCm-4;
H–C C
C(δ=C O= )1a8n7.d9 Hpp-1m1;a Ct3=JO) anwdi tHh-C1′- 2at( δ3JH–=C w15i5th.1 Cp-p2 m(δ;CC =- O15C5H.1 p)pamnd; Ca-tO2JCH3)w anitdh aCt- 23JH(δ–C w=it1h3 1C.9-3p (pδCm =)
H–C C 3 H–C C
(1F3i1g.u9 rpep5mb)). (SFiimguilraer l5yb,t)h. eSismigilnaarllyo,f tHhe-1 s1i1gsnhaolw oef dH3-J1′′ shcoowrreedla 3tJiHo–nCs cworirthelaCt-i4on(δs w=it1h8 C7.-94 p(pδCm =; C18=7O.9)
H–C C
apnpdmC; C-6=(Oδ) a=n1d3 C0.-56 p(δpCm =) 1a3n0d.52 pJpm)w ainthd C2J-H5–C( δwit=h 1C4-85. 3(δpCp =m 1)4(8F.3ig puprem5)b ()F.igure 5b).
C H–C C
CCoommppoouunndd 222:2:T hTeh1eH -1NHM-NRMspRe cstpruemctrsuhmow sshaopwast tear npchatatrearcnt ercihstaircaocftearriosmtica tiocft ria-rsoumbsattiitcu titorni-,
is.ue.b,sdtiotuubtiloent,s ii.ge.n, adlosuabtlδet s=ign6.a9l5s aptp δmH =(J 6=.975. 3ppHmz ,(J1 H= 7,.H3 -H41z), a1nHd, δH-4=′) 6a.n9d0 δ(JH= = 76..490H (zJ ,=1 7H.4, HHz-6, 11),Ha,
H H
dHo-u6′b),l ea ddoouubblleet dsiogunballeatt sδign=al6 a.7t 0δ(HJ == 67..730a (nJ d= 77..43 Hanzd,1 7H.4, HH-z5,1 )1.HT,h He-p5o′)s.i tTiohne opfotshiteiogne roafn tyhlec ghearinanoynl
H
chain on the aromatic ring was established by two-dimensional (2D) HMBC correlations. In this
thearomaticringwasestablishedbytwo-dimensional(2D)HMBCcorrelations. Inthisspectrum,a
2sJpectrcuomup, lai n2JgH-oCf cHou-8pl(iδng =of2 H.78-8– 2(δ.7H4 =, m2.,728H–2).7w4i,t mhC, 2-1H1)( δwit=h 1C2-01.′4 (δpCp =m 1)2a0n.4d pap3mJ ) ancdo uap 3JliHn-Cg cboeutwpleineng
H-C H C H-C
tbInhete.t wJ.s Miegeonnl.a Sltcshi. eo2 0f1sC6ig, -1n271a,a l8ns4 0do fC -C6-12a′atnδdC C=-165′ 1a.t9 δaCn d= 112561.9.9 papnmd, 1re2s6p.9e cptipvmel,y ,rewsepreectoivbesleyr,v ewde.reIn oabdsde1r0itv oieof d1n8.,
cIno rardeldaittiioonns, acotr3rJelatiboentsw aete 3nJHt-hC ebeCtHwe-eAnr tghreo CuHp3(-δAr= gr2o.1u6pp (pδmH =) w2.1it6h pCp-m21)a wndithC -C4-12a′ taδnd =C1-45′1 a.9t δanC d=
H-C 3 H C
11p25r81e..s49e pnacpnemd o,1fr2 e8tsw.p4o ep chptiymvde,lr yro,exasypnled cgatriov2uJelpys, aiwnn idtthh aeC 2g-J3He1-rC(a δwnyit=lh c 1Ch2a6-3i.n2′ ) (wδwCae =sr e1co2ob6n.sf2ei)rr mvweeeddre (bF oyigb usthererev 6oeadb)s. (eTFrhivgeautprioreen 6s eaon)f. c Ttewhooef
H-C C
tptwerreotsiaehrnyycd erc oaoxrfby itlnwgorolo icuh pysidsgrinnoaxtlyhs le aggt reδorCau n=py s7l 2icn.h9 a tahinned wg 7ae1sra.c3no ypnlpfi mcrmh aienidn tbhwyea ts1h3 Ceco oNnbfMsiermrRv easdtpi eobcnytro utfhmtew. ooTbhtseeersrteiva asritygiocnnaa rlobs fiwn towelrioce
staeisgrstniigaanrlsye adct atδrob ci=naor7bl5ioc.8n ssai gnCnd-a67l s1a .an0td pδ pCC m-=2 ,7i nr5e.t8sh paeen1cd3tiC v7e1Nl.y0M , pRbpyms ptw eincot -rtduhiemm 1.3eCTnh sNieosMneaRwl es(2preDeca)t srHsuiMgmnB.e CTdh tceoosrcera ewrlbaeotrineo snaCss.s- i6Tghanunesdd,
C
Cttoh- e2c ,aCrreHbso3pn-Cesc 6Ct igv-6re olayun,pdb ysCht-ow2w,o re-eddsip mceocertnrievsileoalntyiao, lbn(y 2a tDtw 2)JoHH--CdM iwmBiCtehnc scoiaorrrnbeailnal to(i2loiDnc )sc .HaTrMbhouBnsC, a Hcto -Cr8r-6seh l(aoδtwCio =en d7s2.c T.o9hr pruepsl,am Hti)o,- 8nw sahhtiol3ewJ Hteh-dCe
wcmoiertrthhelcyaalt rigborinon uaoptl is3cJ aHct-aC δr wbC o=itn 3h1a c.t2aC rab-n6idn( oδ2lC9ic.9= c pa7pr5bm.8o np(C paHtm C3)-,1-6w a (nhδdeCr C=e aH7s53t-.8hC e2p,mp rmeest)ph, eywclthgievrroeeluyaps) ssthhaoet wδmCeed=t hc2yo9lr .r2gerlpoaputimposn (saC taH tδ 23CJ- H=1-
a2C n9w.d2i tCphpH Cm3--2 C( C(2δH)Cs 3=h- 1o7 w1a.n3ed dp CpcoHmr3r)-e C(lFa2it)gi ousnhrseo wa6tbe2)d.J Hc-oCrrweliathtioCn-s2 a(δt C2JH=-C7 1w.0ithp pCm-2) ((δFCig =u 7r1e.60a p).pm) (Figure 6a).
Compound 23: A similar analysis was conducted to elucidate the structure of this compound.
The 1H-NMR spectrum shows a pattern characteristic of aromatic tri-substitution, i.e., a singlet signal
at δH = 6.95 ppm (1H, H-2′, meta-coupling of H-2′ with H-6′ was not detected), a doublet signal at
δH = 6.90 (J = 8.1 Hz, 1H, H-6′) and a doublet signal at δH = 6.68 (J = 8.1 and 1H, H-5′). The position of
the geranyl chain on the aromatic ring was established by two-dimensional (2D) HMBC correlations.

In this spectrum, a 2JH-C coupling of H-8 (δH = 2.65–2.52, m, 2H) with C-1′ (δC = 135.5 ppm) and a 3JH-C
coupling bFFeiigtgwuurreeee 66n.. tMMheaa iisnni g22DnDa HHlsMM oBBf CCC -oo2bb′asseenrrdvv eeCdd- 6ccoo′ rarrrtee δllaaCtt i=ioo 1nn3ss 0ffoo.9rr ::a ((naa)d) CC 1oo2mm6.pp7oo puupnnmdd 22, 22r;e; (s(bbp))e CCctooimmvepplooyuu, nnwdde 22r3e3.. o bserved.
In addition, correlations at 3JH-C between the CH3-Ar group (δH = 2.22 ppm) with C-2′ and C-4′ at δC =
130.9CB aeoncmdau p1so5eu1 .nC6d opm2p3pm:oA,u rnesdsimp e2icl4at irwvaeanlsya ,lo yabsntidasi nwae a2dJsH -cfCro ownmdit uh1c 7Cte -bd3y′t (otδheCe l=u hc1iy2dd3ar.t4ae)t itwohnee rsrete roaubcctstieournrve eoodff (itFth,i igstuhcreoe m a6rbpo)om. uTanhtdiec.
Tsuhbes1tHitu-NtioMnR psaptteecrtnru wmass hmoawinstaaipnaetdte fronr cChoamrapcoteurnisdtisc 2o4f aanrodm 2a5t. iTchtrui-ss, uinb sCtiotumtpioonu,ni.de. ,2a4,s tihneg lpertessiegnncael

aotf δtwo= 6h.y9d5rpopxmyl (g1rHou,Hps- 2i1n, mtheet ag-ceoraunpyliln cghoafinH w-2a1sw citohnfHir-m61ewd absyn othted eotbescetrevda)t,iaond oouf btlwetos itgenrtailarayt
H
δcarb=in6.o9l0ic( Jsi=gn8a.1lsH azt ,δ1CH =, 7H5-.86 1a)nadn d70a.9d opupbmle itns itghnea 1l3Cat NδM=R 6s.p6e8c(tJru=m8..1 Tahnedse1 sHig,nHa-l5s1 )w. Terhee apsosisgitnioend otof
H H
carbons C-3′ and C-7′, respectively, by two-dimensional (2D) HMBC correlations. Thus, the CH3-C7′
and CH3-8′ groups showed correlation at 2JH-C with carbinolic carbon at C-7′ (δC = 70.9 ppm), and
therefore, the signal at δC = 75.8 ppm was unequivocally assigned to C-3′ (Figure 7a); while for
Compound 25, the methylene group (at δC = 22.7 ppm, assigned as C-7′) showed a correlation at 2JH-C
with C-2 (δC = 114.2 ppm) and C-1′ (δC = 48.3 ppm) and 3JH-C with a tertiary carbinolic carbon at
δC = 76.7 ppm assigned to C-2′. Additionally, the CH3-C2′ group at δH = 1.20 ppm (3H, s) showed
coupling at 2JH-C with C-2′ and 3JH-C with tertiary C-1′ (δC = 48.3 ppm) (Figure 7b) Thus, the cyclohexane
structure is confirmed for geranyl chain. Finally, mono-hydroxylation in the geranyl chain for
Compound 26 was mainly established by 13C NMR data and 2D HMBC correlations. Only one signal
of carbinolic carbon at δC = 70.9 ppm in the 13C NMR spectrum was observed, and the methyl groups
at δH = 1.22 (6H, s, CH3-C7′ and H-8′) showed 2JH-C correlations with this carbon (C-7′, δC = 70.9 ppm)
(Figure 7c). In addition, these methyl groups showed 3JH-C correlations with C-6′ (δC = 43.3 ppm)
(Figure 7c).

Figure 7. Major 2D HMBC observed correlations for Compounds 24 (a), 25 (b) and 26 (c).
Int. J. Mol. Sci. 2016, 17, 840 10 of 18
presence of two hydroxyl groups in the geranyl chain was confirmed by the observation of two
tertiary carbinolic signals at δC = 72.9 and 71.3 ppm in the 13C NMR spectrum. These signals were
aInsts.iJg.Mneodl. Stcoi. 2c0a1r6b,o17n,s8 C40-6 and C-2, respectively, by two-dimensional (2D) HMBC correlations. 1T0houf1s8,
the CH3-C6 group showed correlation at 2JH-C with carbinolic carbon at C-6 (δC = 72.9 ppm), while the
mtheethgeyrl agnryolucphsa aint δoCn =t 3h1e.2a raonmda 2t9ic.9r ipnpgmw (aCsHes3-t1a balnisdh CedHb3-yCt2w, roe-sdpimecetinvseiloyn)a slh(o2wDe)dH cMorBrCelactoirornesla atti o2JnHs-.
CIn wtihtihs Csp-e2c (tδruC m= 7,1a.23J ppmc)o u(Fpilginugreo 6fbH).- 8(δ =2.65–2.52,m,2H)withC-11(δ =135.5ppm)anda3J
H-C H C H-C
couplingbetweenthesignalsofC-21andC-61atδ =130.9and126.7ppm,respectively,wereobserved.
C
Inaddition,correlationsat3J betweentheCH -Argroup(δ =2.22ppm)withC-21 andC-41 at
H-C 3 H
δ =130.9and151.6ppm,respectively,anda2J withC-31(δ =123.4)wereobserved(Figure6b).
C H-C C
Thepresenceoftwohydroxylgroupsinthegeranylchainwasconfirmedbytheobservationoftwo
tertiarycarbinolicsignalsatδ =72.9and71.3ppminthe13CNMRspectrum. Thesesignalswere
C
assignedtocarbonsC-6andC-2,respectively,bytwo-dimensional(2D)HMBCcorrelations. Thus,the
CH -C6gFrioguupres 6h. oMwaeind 2cDo rHreMlaBtCio onbasetr2vJed cowrrietlhatcioanrsb finoro: l(iac) cCaormbopnouantdC 2-26; ((δb) C=om72p.9oupnpdm 23),. whilethe
3 H-C C
methylgroupsatδ =31.2and29.9ppm(CH -1andCH -C2,respectively)showedcorrelationsat
C 3 3
Because Compound 24 was obtained from 17 by the hydration reaction of it, the aromatic
2J withC-2(δ =71.3ppm)(Figure6b).
H-C C
substitution pattern was maintained for Compounds 24 and 25. Thus, in Compound 24, the presence
Because Compound 24 was obtained from 17 by the hydration reaction of it, the aromatic
of two hydroxyl groups in the geranyl chain was confirmed by the observation of two tertiary
substitutionpatternwasmaintainedforCompounds24and25.Thus,inCompound24,thepresenceof
ctwarobihnyodlirco sxiyglngarlos uapt sδCin = t7h5e.8g earnadn y70l.c9h paipnmw ians tchoen 1fi3Crm NeMdRby sptheectorbusmer. vTahtieosne osifgtnwaolst weretiraer yascsaigrbniendo ltioc
csaigrbnoalnssa Ct-δ3′ a=n7d5 .C8-a7n′, dre7s0p.e9cptipvmelyin, btyh etw13oC-dNimMeRnssipoencatlr u(2mD.) THhMesBeCsi cgonrarleslawtieornesa. sTshigunse, dthteo CcaHr3b-oCn7s′
C
aCn-d31 CanHd3-8C′ -g71r,oruepssp eschtoivweelyd, bcoyrrtwelaot-idoinm aetn 2sJiHo-nC awli(t2hD c)aHrbMinBoClicc coarrrbeloanti oant sC.-7T′h (uδsC, =th 7e0.C9 Hpp-mC7),1 aanndd
3
tChHere-f8o1rger,o tuhpes ssihgonwale datc oδrCr e=la 7ti5o.n8 aptp2mJ wwasi thuncaerqbuiinvoolcicacllayr baosnsiagtnCed-7 1t(oδ C=-37′ 0(.F9igpuprme ),7aan);d wthheirleef oforer,
3 H-C C
Cthoemspigonuanlda t25δ, th=e 7m5.e8thpyplmenew garsouupn e(qaut δivCo =c a2l2l.y7 apspsmig,n aesdsitgoneCd-3 a1s( CFi-g7u′)r seh7oaw);ewd ha icleorfroerlaCtioomn pato 2uJnH-dC
C
w25i,thth Ce-m2 e(tδhCy =le n11e4g.2ro puppm(a) taδnd= C2-21.′7 (pδpC m= ,4a8s.s3i gpnpemd)a asnCd- 731J)Hs-Ch owwitehd aa tceorrtiraerlyat icoanrbaitn2oJlic cwaribtohnC a-2t
C H-C
δ(δC == 7161.74 .p2ppmpm a)sasingdneCd-1 t1o( δC-2=′.4 A8.d3dpiptimon)aalnlyd, 3tJhe CwHi3t-hCa2′t egrrtoiaurpy caat rδbHin =o l1ic.2c0a rpbpomn a(t3δH, =s)7 s6h.7opwpemd
C C H-C C
caosusipglninedg atot 2CJH--2C1 w. Aitdhd Cit-i2o′ naanldly 3,JtHh-Ce wCiHth -tCer2t1iagrryo Cup-1a′ t(δδC = =481..32 0pppmpm) (F(3igHu,rse) 7sbh)o Twheuds,c tohuep clyincgloahte2xJane
3 H H-C
swtriuthctCu-r2e1 aisn dco3nJfirmwedit hfoterr tgiaerryanCy-l1 1ch(δain=. 4F8in.3apllpy,m m)(oFnigou-hryed7rbo)xTyhlautsio,nth einc ytchleo hgeexraanneyls tcruhcatiun refoirs
H-C C
Compound 26 was mainly established by 13C NMR data and 2D HMBC correlations. Only one signal
confirmedforgeranylchain. Finally,mono-hydroxylationinthegeranylchainforCompound26was
omf acianrlbyinesotlaicb lciashrbeodnb ayt 1δ3CC =N 7M0.9R pdpamta iann tdh2eD 13CH MNMBCR csoprerectlarutimon ws.aOs nolbysoenrveesdig, nanaldo tfhcea mrbeinthoylilc gcraorubopns
aatt δδH == 17.02.29 (p6pHm, si,n CtHhe3-C137C′ aNnMd RH-s8p′e) csthrouwmewd a2JsHo-Cb csoerrrveeldat,iaonnds wthiethm tehtihs yclagrbroounp (sCa-7t′δ, δC= = 17.02.29 (p6pHm,s),
C H
(CFHigu-rCe7 71ca)n. dInH a-d81d)istihoonw, ethde2sJe mectohryrle lgartoiounpss wshitohwthedis 3cJaHr-Cb ocnor(rCel-a7t1i,oδns =w7i0th.9 Cp-p6m′ ()δ(CF =ig 4u3r.e3 7pc)p.mIn)
3 H-C C
(Figure 7c).
addition,thesemethylgroupsshowed3J correlationswithC-61(δ =43.3ppm)(Figure7c).
H-C C

Figure7.Major2DHMBCobservedcorrelationsforCompounds24(a),25(b)and26(c).
Figure 7. Major 2D HMBC observed correlations for Compounds 24 (a), 25 (b) and 26 (c).

2.3. InVitroAntifungalActivityagainstB.cinerea.
Allstudiedcompounds(14–26)weretestedforinvitroantifungalactivityonthemycelialgrowth
ofB.cinereastrainGM7usingtheagarradialassaywithPotatoDextroseAgar(PDA).Figure8showsan
assaywheretheB.cinereamyceliumgrowsinmediumcontainingonlyPDAand1%ethanol(Figure8a,
negativecontrol), Captanat250ppm(Figure8b, usedinthisstudyasapositivecontrol)andtwo
differentconcentrationsofCompound26(Figure8c,150ppm;Figure8d,250ppm).