Broad-spectrumnon-toxicantiviralnanoparticleswithavirucidalinhibitionmechanismValeriaCagno,PatriziaAndreozzi,MarcoD’Alicarnasso,PauloJacobSilva,MarieMueller,MarieGalloux,RonanLeGoffic,SamuelT.Jones,MartaVallino,JanHodek,JanWeber,SoumyoSen,Emma-RoseJaneček,AhmetBekdemir,BarbaraSanavio,ChiaraMartinelli,ManuelaDonalisio,Marie-AnneRameixWelti,Jean-FrancoisEleouet,YanxiaoHan,LaurentKaiser,LelaVukovic,CarolineTapparel,PetrKrál,SilkeKrol,DavidLembo&FrancescoStellacci

NatureMaterialsvolume17,pages195–203(2018)

DipartimentodiScienzeClinicheeBiologiche,UniverisitàdegliStudidiTorino,Orbassano,Italy- ValeriaCagno,DavidLembo

InstituteofMaterials,EcolePolytechniqueFédéraledeLausanne(EPFL),Lausanne,Switzerland- ValeriaCagno,MarieMueller

SITorino

SSTorino

SSSestoFiorentino

SSPortici

SSBari

Broad-spectrumnon-toxicantiviralnanoparticleswithavirucidalinhibitionmechanismValeriaCagno,PatriziaAndreozzi,MarcoD’Alicarnasso,PauloJacobSilva,MarieMueller,MarieGalloux,RonanLeGoffic,SamuelT.Jones,MartaVallino,JanHodek,JanWeber,SoumyoSen,Emma-RoseJaneček,AhmetBekdemir,BarbaraSanavio,ChiaraMartinelli,ManuelaDonalisio,Marie-AnneRameixWelti,Jean-FrancoisEleouet,YanxiaoHan,LaurentKaiser,LelaVukovic,CarolineTapparel,PetrKrál,SilkeKrol,DavidLembo&FrancescoStellacci

NatureMaterialsvolume17,pages195–203(2018)

DipartimentodiScienzeClinicheeBiologiche,UniverisitàdegliStudidiTorino,Orbassano,Italy- ValeriaCagno,DavidLembo

InstituteofMaterials,EcolePolytechniqueFédéraledeLausanne(EPFL),Lausanne,Switzerland- ValeriaCagno,MarieMueller

Sviluppodinanoparticelleconazionevirucidaadampiospettro

Infezionivirali

Prevenzione:vaccini

Dopol’infezione:farmaciantivirali numerolimitatovirus-specificiazioneintracellulare(replicazionevirale)tossicitàintrinseca

Sostanzevirustatiche azioneextracellulareampiospettronontossiche

Legametraviruserecettorecellulare(comuneapiùvirus)

VAL(viralattachmentligand)<===>HSPG(HeparansulfateProteoglycans)Multiplebasicaminoacid caricanegativadelgrupposolfato

legamereversibile=>inutilicomefarmaci

Virustatico

Shortlinker

MercaptoetilsulfonatoMES

Virucida???

Longlinker

MercaptoundecanosulfonatoMUS

GliattualivirustaticimimanolastrutturadiHSPGShortlinkers=>legameapocheunitàripetutedelVAL=>legamedeboleereversibile

Farmacoideale: virucida=>inattivazioneirreversibileampiospettroNontossico

10

b,Heparin,MES-NPsandMUS:OT-NPsviralinfectivitycurvesandvirucidalassays.Thepercentagesofinfectionwerecalculatedbycomparingthenumberofplaquesintreatedanduntreatedwells.c,VirucidalactivityofMUS-coatedNPsagainstHPV-16,RSV,LV-VSV-G(indicatedasLV)andDENV-2viruses.d,MUS:OT-NPsinhibitionofviralinfectivityagainstHSV-2versustime(minutes).

InfectivityupondilutionCelltoxicity

Infectivity

a–c,SamplesimagedusingdrynegativelystainedTEM(a)orunstainedcryo-TEM(b,c).Thescalebarsare100 nm.

HSV-2herpessimplexvirus2

d,PercentageanddistributionofNPs(MESorMUS:OT)associatedwithHSV-2immediatelyandafter90 min

SupplementaryFigure11:VirucidalAssaysandInteractionwithHSV-2virusesforEG2-OH-NPs.A)Cryo-TEM(up)andnegativestainingTEM(down)imagesofHSV-2virus(105pfu)treatedwithEG2-OH-NPs.B)VirucidalassayofHSV-2virustreatedwithEG2-OH-NPs.n=3C)%distributionofnotsulfonatedEG2-OHNPsassociatedwithHSV-2after1.5hofincubation.ThemajorityofviruseswerenotassociatedwithanyNP.Thedistributionwascalculatedanalysingbetween50and100images.Thescalebarsindicate100nm

SupplementaryFigure22:Cryo-TEMandTEMofHSV-2withenvelope.A)Cryo-TEMandB)negativestainingTEMimagesofHSV-2virus(105pfu)treatedwithNPs.(Scalebar100nm)

SupplementaryFigure12:MUS:OT-NPsassociatedwithHPV-16PsVs.Intheupperpaneltheimageswereacquiredbycryo-TEM.Inthepanelbelowthesampleswerestainedwithuranylacetate0.5%for5minandacquiredbyTEM.ImagesaanddrepresenttheHPV-16,whileb,c,eandfrepresentdifferenttypesofNPsassociationwithHPV-16PsVs.Thescalebarsindicate100nm.

HPVhumanpapillomavirus

a,CartoonofthevirucidalactivityofMUS:OT-NPscomparedtoMES-NPs.

RSVrespiratorysyncytialvirus

humancervicovaginalhistocultures EpiVaginaltissues

PhilipsCM10