Syn hesis o gian globula mul i alen glyco ulle enes as po en
inhibi o s in a model o Ebola i us in ec ion
An onio Muñoz,§ Da id Sigwal ,¥,£ Bea iz M. Illescas,§ Joanna Luczkowiak,‡ Lau a
Rod íguez,§ Iwona Nie enga en,¥ Michel Holle ,¥ Jean-Se ge Remy,£ Ke in Bu e ,¶
S éphane P. Vincen ,¶ Ja ie Rojo,†,* Ra ael Delgado,‡,* Jean-F ançois Nie enga en,¥,*
Naza io Ma ín§,,*
§Depa amen o de Química O gánica, Facul ad de Química, Uni e sidad Complu ense, 28040
Mad id, Spain, e-mail: [email p o ec ed]s; † Glycosys ems Labo a o y, Ins i u o de
In es igaciones Químicas (IIQ), CSIC – Uni e sidad de Se illa, A . Amé ico Vespucio 49, Se ille
41092 Spain. Tel: + 34 954489568; FAX +34 954460165; e-mail: ja ie[email p o ec ed]sic.es;
£ Labo a o y V-SAT (CAMB UMR 7199, CNRS), Labex Medalis, Uni e si é de S asbou g, 74
Rou e du Rhin, 67401 Illki ch-G a ens aden, F ance; ¶ Uni e si y o Namu (FUNDP),
Dépa emen de Chimie, Labo a oi e de Chimie Bio-O ganique, ue de B uxelles 61, B-5000
Namu , Belgium; ‡Labo a o io de Mic obiología Molecula , Ins i u o de In es igación Hospi al
12 de Oc ub e (imas12) 28041 Mad id, Spain, e-mail: a [email protected] id.o g;
¥ Labo a oi e de Chimie des Ma é iaux Moléculai es, Uni e si é de S asbou g e CNRS (UMR
7509), Ecole Eu opéenne de Chimie, Polymè es e Ma é iaux, 25 ue Becque el, 67087
S asbou g, F ance, e-mail: nie enga en@unis a. ;

IMDEA-Nanoscience, Campus
Can oblanco, 28049 Mad id, Spain.
Abs ac
The use o mul i alen ca bohyd a e compounds o block cell-su ace lec in ecep o s is a
p omising s a egy o inhibi ing he en y o pa hogens in o cells and could lead o he
disco e y o no el an i i al agen s. One o he main p oblems wi h his app oach, howe e , is
he di icul y in making compounds wi h adequa e size and mul i alency o mimic na u al
sys ems such as i uses. Hexakis-adduc s o [60] ulle ene a e a use ul building block in his
ega d because hey main ain a globula shape while allowing con ol o e size and
mul i alency. He e we epo wa e -soluble ideca ulle enes deco a ed wi h 120 pe iphe al
ca bohyd a e subuni s, so-called 'supe balls', ha can be e icien ly syn hesized om hexakis-
adduc s o [60] ulle ene in one s ep by using coppe -ca alyzed azide–alkyne cycloaddi ion
(CuAAC) click-chemis y. In ec ion assays show ha hese 'supe balls' a e po en inhibi o s o
cell in ec ion by an a i icial Ebola i us wi h IC50s in he sub-nanomola ange.
Mul i alency is a gene al and e icien ool used by na u e o achie ing s ong in e ac ions in a
e e sible manne . A he molecula le el, mul i alen in e ac ions ha e he ad an age o
enhancing d as ically he binding be ween molecules when compa ed wi h mono alen
binding.1
A ema kable example om na u e whe e mul i alency plays a signi ican ole is he
in e ac ion be ween i uses and bac e ia wi h hei espec i e hos cells. In pa icula , DC-SIGN
(dend i ic cell-specific in e cellula adhesion molecule-3-g abbing non-in eg in) ecep o is one
o he mos impo an pa hogen ecogni ion ecep o s. This lec in e icien ly ecognizes in a
mul i alen manne saccha ides con aining mannoses and ucoses om glycop o eins.2 In his
ega d, i is well es ablished ha p o ein−ca bohyd a e in e ac ions a e a key issue in a a ie y
o biological p ocesses, bu since he affini y o simple glycans o hei espec i e ecep o s is
o en weak, mul i alen in e ac ions ypically occu . An impo an open challenge o esea ch
nowadays is he be e unde s anding and p ac ical use o mul i alency. Ac ually, in a b oade
sense, glycobiology is cu en ly a ield o esea ch whe e chemically inspi ed app oaches and
s a egies a e p oducing signi ican ad ances.3
I is well-known, howe e , ha some i uses a e able o escape om p ocessing by he
immune de ense by using DC-SIGN as an en y poin o in ec he cell. The e o e, inhibi ing he
en y o pa hogens by blocking his ecep o a he ea ly s ages o in ec ion ep esen s a
aluable s a egy o he design o new an i i al agen s. In o de o add ess his challenge, a
a ie y o di e en mul i alen sca old a chi ec u es ha e been syn hesized, all o hem
endowed wi h mul iple ca bohyd a es.4 Design o hese glyco-conjuga es ypically equi es a
mul i alen cen al sca old o co e co alen ly connec ed o he ca bohyd a e epi opes
deco a ing he pe iphe y. Thus, glyco-clus e s in which he ca bohyd a e uni s a e di ec ly
connec ed o he co e, glyco-dend ime s connec ed h ough a dend i ic s uc u e, and glyco-
polyme s in ol ing a polyme ic backbone ha e been in ensi ely in es iga ed in ecen yea s.5-9
Fulle enes ha e also been employed as a biocompa ible sca old o he mul i alen
p esen a ion o ligands, gi en he possibili y o mul iple unc ionaliza ion on hei con ex
su ace. In pa icula , hexakis-adduc s o [60] ulle ene wi h a Th-symme ical oc ahed al
addi ion pa e n ha e a unique h ee-dimensional s uc u e which allows he in oduc ion o
six one- ype o mixed- ype addends.10,11 Such de i a i es can be ob ained in one syn he ic s ep
by he addi ion o malona es o C60, bu his app oach has been limi ed by he low yields ha
esul when la ge malona es a e used because he eac ion is e y sensi i e o s e ic e ec s.12
We ha e ecen ly de eloped a p ocedu e based on a click-chemis y app oach which p o ides
hexakis-adduc s wi h wel e alkyne o azide e minal g oups in high yields om simple
malona es.13,14 These hexakis-adduc s can be easily and e icien ly unc ionalized by using he
coppe -ca alyzed azide–alkyne cycloaddi ion (CuAAC) eac ion allowing he in oduc ion o
wel e unc ional g oups simul aneously in a egioselec i e and e icien way. Since hese
hexakis-adduc s show an oc ahed al a angemen o equally sepa a ed addends loca ed on he
[60] ulle ene pe iphe y in a globula opology, hey cons i u e a e y a ac i e pla o m o
he s udy o mul i alen in e ac ions wi h lec ins, wi h impo an ad an ages such as he
be e biocompa ibili y o he ca bon cen al co e, he globula geome y o he unc ional
g oups and he ease and e sa ile chemical unc ionaliza ion.
We ha e ecen ly epo ed he syn heses o [60] ulle ene hexakis-adduc s endowed wi h
di e en ca bohyd a e uni s in he pe iphe y. Depending upon he s a ing suga de i a i es
employed (monome , dime o ime ) 12, 24 and up o 36 monosaccha ides ha e been
in oduced on he pe iphe y o he ulle ene cen al co e in a s aigh o wa d manne .14-17
Some o he a o emen ioned glyco ulle enes ha e shown in e es ing bio-medical
applica ions.18 A signi ican mul i alen e ec has been obse ed in he inhibi ion p o ile o a
[60] ulle ene hexakis-adduc endowed wi h 12 iminosuga uni s owa ds di e en
glycosidases.16 On he o he hand, ulle ene hexakis-adduc s bea ing 12 mannoses on he
pe iphe y beha e as inhibi o s o FimH, a bac e ial adhesin.19 Fu he mo e, s udies ca ied ou
on hexakis-adduc s o [60] ulle ene endowed wi h 12, 24 and 36 mannoses ha e shown a
mul i alen e ec in he in e ac ion wi h concana alin A,15 and ac as e icien inhibi o s o cell
in ec ion by Ebola pseudo yped i al pa icles.20 These p elimina y s udies e eal ha
ulle enes a e adequa e pla o ms o he mul i alen p esen a ion o ca bohyd a es, hus
pa ing he way o he co alen linkage o a wide a ie y o di e en bio-ac i e molecules in a
mul i alen manne and a singula and less-explo ed globula opology.
In his wo k, we ha e syn hesized molecules wi h a globula opology o med by an hexakis-
adduc o hexakis adduc s o [60] ulle ene, ca ying ou he as es dend ime ic g ow h
epo ed up o now, wi h he in oduc ion o 120 suga uni s in one syn he ic s ep by using he
highly e icien CuAAC click-chemis y me hodology. The syn hesis o hexakis-adduc s o
hexakis-adduc s o [60] ulle ene wi h an oc ahed al addi ion pa e n has p e iously been
epo ed. This p e ious example used mac ocyclic bis-malona es and enabled he syn hesis o
hep a ulle enes wi h unable p ope ies.21-23 In con as , ou click-chemis y app oach yields
ideca ulle enes in which he cen al [60] ulle ene is co alen ly connec ed o wel e
[60] ulle enes, each o hem endowed, in u n, wi h en monosaccha ides. To con i m he
p oposed s uc u es, we ha e ca ied ou a ho ough s uc u al s udy in ol ing a a ie y o
echniques (In a ed, 1H NMR and 13C NMR spec oscopy; dynamic ligh sca e ing (DLS),
ansmission elec on mic oscopy (TEM) and X- ay pho oelec on spec oscopy (XPS)).
Biological s udies e eal ha he new gian glyco ulle enes exhibi a e y s ong inhibi ion o
cell in ec ion by Ebola pseudo yped i al pa icles wi h IC50 alues in he sub-nanomola ange.
Syn hesis
The syn hesis o he glyco ulle ene supe balls has been ca ied ou as depic ed in Figu es 1-3.
Al hough hese molecules (17a-c) may appea di icul o ob ain a a i s glance, mainly due o
hei size and molecula complexi y, hei syn hesis is s aigh o wa d and, mos impo an ly,
easily ep oducible a o ding he inal compounds in good o e all yields. Fu he mo e, as
discussed below, pu i ica ion o he samples — including he equi ed emo al o coppe o
u he biological s udies — a e ca ied ou easily because a e column il a ion hese
compounds (17a,b) a e p ecipi a ed in he eac ion medium and pu e samples showing highly
ep oducible biological assays a e ob ained.
Wi h he aim o s udying he e ec o he size and/o s e ic conges ion o hese supe balls on
he biological p ope ies, wo di e en and complemen a y syn he ic s a egies ha e been
simul aneously de eloped o ob ain inal p oduc s wi h di e en space s inbe ween he
cen al ulle ene co e and he pe iphe al ca bohyd a e-subs i u ed ulle ene appendages. In
bo h cases, he syn hesis elies on he g a ing o clickable A10B mac omonome s on o a
compac C60 hexa-adduc sca old bea ing wel e e minal alkyne uni s. The p epa a ion o
mac omonome 9 is depic ed in Figu e 1. Es e i ica ion o alcohol 1 wi h e hylmalonyl chlo ide
ollowed by eac ion o he esul ing malona e (2) wi h C60 unde Bingel condi ions ga e
me hano ulle ene 3. Subsequen ea men o 3 wi h an excess o malona e 4, CB 4, and DBU
in o-dicho obenzene (ODCB) ga e building block 5 o which mannose de i a i e 6 was clicked.
Impo an ly, he TMS-p o ec ed alkyne uni is no eac i e unde hese condi ions and
in e media e 7 was hus ob ained in a good yield. Finally, compound 7 was desilyla ed in si u
wi h e abu ylammonium luo ide (TBAF) o gene a e he co esponding e minal alkyne and
eac ion wi h a la ge excess o diazide 8 p o ided he desi ed mac omonome 9 in 87% yield.
The syn hesis o mac omonome s 15a-b s a s om monoadduc 10 esul ing om he Bingel
nucleophilic cyclop opana ion o C60 wi h 6-b omohexyl e hyl malona e (Figu e 2).24 To ob ain
he [5:1]-hexaadduc 12, a en old excess amoun o di(pen -4-yn-1-yl) malona e and a 50-
old excess o ca bon e ab omide we e added in he p esence o DBU as he base. Flash
ch oma og aphy pu i ica ion p o ided hexaadduc 12 as a ed solid. Di e en ca bohyd a e

azides (mannose, galac ose) we e hen linked o C60 by he CuAAC eac ion, employing
CuB ·S(CH3)2 as he ca alys and sodium asco ba e as he educing agen in he p esence o a
piece o me allic Cu, o yield de i a i es 14a-b. The nucleophilic subs i u ion o b omine by
azide was ca ied ou wi h an excess o sodium azide unde mic owa e hea ing, hus gi ing
e sa ile building blocks 15a-b.
The azide-con aining mac omonome s 9 and 14a-b we e hen clicked o symme ic alkyne
de i a i e 1614 unde CuAAC condi ions. Supe balls 17a-c subs i u ed wi h up o 120
monosaccha ide uni s we e hus ob ained wi h yields o e 70% (Figu e 3). De i a i es 17a-b
we e ob ained in only h ee syn he ic s eps om easily accessible eac i es and a oiding he
use o p o ec ing g oups.
Al hough he h ee ideca ulle enes 17a-c con ain he same numbe o monosaccha ides,
compound 17c has a la ge space be ween he cen al ulle ene moie y and he pe iphe al
ca bohyd a e unc ionalized ulle enes. This longe space a ec s he lexibili y and he size o
he compound and can a ou he accessibili y and a ailabili y o he ca bohyd a e ligands o
in e ac wi h he ecep o , which could ha e an impo an in luence on he biological ac i i y.20
I is impo an o no e, howe e , ha he a o emen ioned syn he ic app oaches enables
ulle ene de i a i es o be syn hesized om malona es endowed wi h alkyl chains o a iable
leng h h ough a Bingel cyclop opana ion ollowed by a subsequen CuAAC eac ion. This
syn he ic s a egy a o ds hexakis-adduc s wi h long chains connec ing he ulle enes and
ca bohyd a es since hei di ec p epa a ion om sui ably unc ionalized malona es bea ing
long chains ypically occu wi h low yields.10
Cha ac e iza ion o hese supe balls was ca ied ou by s anda d spec oscopic echniques.
Thus, FTIR spec a do no show he p esence o ei he azide g oups ( ypical signal obse ed a
2097 cm-1) o alkynes ( 2117 cm-1) (See he Supplemen a y In o ma ion). The molecula ion
peak o hese compounds could no be de ec ed bu i should be no ed ha he ans e o
such high molecula weigh glycoclus e s in o he gas phase du ing MALDI-TOF MS analysis is
e y di icul . Mo eo e , bo h he suga s and he ulle ene hexaadduc moie ies gi e ise o a
high le el o agmen a ion.25 The unambiguous s uc u al cha ac e iza ion o 17a-c was,
howe e , g ea ly acili a ed by hei high symme y. Indeed, 13C NMR spec oscopy was
pa icula ly help ul o he cha ac e iza ion o hexakis-adduc s o [60] ulle ene, as only wo
signals a e usually obse ed o he sp2 ca bons o C60, p o iding e idence o he oc ahed al
symme y o he ulle ene co e. As a ypical example, he 13C NMR spec um o compound 17a
is depic ed in Figu e 4. In his case, only wo sp2 ca bons a e obse ed in he spec a (δ  145.4
and 141.6 ppm), whe e we can also dis inguish he wo di e en kinds o iazole ings p esen
in he compound (a δ  146.9 and 123.3 ppm o he ca bons o he ou e iazole ings and a
δ  146.4 and 122.9 ppm o he ca bons o he inne iazole ings). In addi ion, only one
signal is de ec ed o all he ca bonyl g oups (δ  163.8 ppm) and he sp3 ca bons o he C60 (δ
 69.3 ppm), while he malona e b idgehead ca bons p esen in he s uc u e a e obse ed a
δ  45.2 ppm.
Addi ional cha ac e iza ion was achie ed by DLS analysis (H2O, 0.01 mg/mL and 0.1 mg/mL),
whe e, ega dless he concen a ion used, we ound wo o h ee main size dis ibu ions o
17a-c (Supplemen a y Figu e 1). This is compa ible wi h a weak agg ega ion o 17a-c in wa e .
The i s , a ound 5-6 nm, mus co espond o only one molecule, while he second, a 120-
150 nm and hi d, 200 nm, show he agg ega ion o se e al molecules. In DMSO (0.1 mg/mL)
(Supplemen a y Figu e 2), al hough mos o he molecules show no agg ega ion, he p esence
o agg ega es o di e en sizes and, especially, e y la ge agg ega es, is also de ec ed. The
endency o o m agg ega es was also con i med by he b oadening o he 13C NMR spec um
eco ded in D2O when compa ed o he one eco ded in DMSO-d6 (see Supplemen a y
In o ma ion).
The TEM images o eshly p epa ed samples e eal he p esence o small sphe ical pa icles,
co esponding o a ew o e en jus one molecule (4 nm), independen ly o whe he he
concen a ion is 0.01 mg/mL o 0.1 mg/mL, in good ag eemen wi h he expe imen al indings
in DLS analyses (Figu e 5 and Supplemen a y Figu e 3).
X- ay pho oelec on spec oscopy (XPS) was addi ionally used o con i m he composi ion o
he suga balls. This su ace echnique enables he iden i ica ion o he a oms p esen on he
molecule oge he wi h hei chemical s a e and hei ela i e abundance. The su ey spec a
o supe ball 17a (Supplemen a y Figu e 4) displays he C 1s, O 1s and N 1s ea u es as
expec ed, wi h no addi ional spec oscopy signa u es o possible impu i ies. Mo eo e , he
high esolu ion N 1s co e le el spec um (Supplemen a y Figu e 4, inse igh ) was composed
o wo di e en componen s wi h a 1:2 a io o he in eg a ed a eas, he one loca ed a 400.3
eV is ela ed o one ni ogen a om o he iazole ing (N–N–N) and he o he cen e ed a
398.8 eV is a ibu ed o he o he wo ni ogen a oms a ached o ca bon a oms (C–N).26 The
absence o a well- esol ed peak a ound 405.0 eV demons a es he lack o he elec on-
de icien ni ogen o he azide g oup in he inal compound.27,28 The composi ion o
compounds 17b and 17c has also been asce ained by hei espec i e XPS analyses which
showed he p esence o he expec ed elemen s, acco ding o hei ela i e abundance (see
Supplemen a y Figu e 4 and Supplemen a y Table 1).
Biological s udies
A numbe o molecules, including DC-SIGN, ha e been p oposed as ecep o s o he Ebola
i us 29-31. Al hough DC-SIGN is no he main ecep o in case o Ebola i us, i is hough o play
a signi ican ole in he cell en ance o his in ec ious agen in signi ican cell popula ions such
as dend i ic cells,29,32 hus acili a ing ea ly i al dissemina ion. The e o e, DC-SIGN can
unc ion as a good model o (i) s udying he i s s eps o pa hogenesis o Ebola i us and (ii)
sc eening he an i i al s a egies based on DC-SIGN- a ge ing compounds o p e en ion and
ea men pu poses. DC-SIGN ecognizes mannosyla ed and ucosyla ed oligosaccha ides
p esen ed in a mul i alen manne on he su ace o se e al pa hogen en elope glycop o eins.
Thus he p epa a ion o mul i alen ca bohyd a e sys ems is necessa y o he e icien
in e ac ion wi h his ecep o as well as o he e ec i e compe i ion wi h he na u al ligands.
In his s udy we ha e e alua ed he inhibi o y e ec o gian globula mul i alen
glyco ulle enes in he expe imen o di ec in ec ion o Ju ka cells exp essing he su ace
ecep o DC-SIGN (Ju ka -DC-SIGN wi h pseudo yped i al pa icles p esen ing Ebola i us
glycop o ein GP1. These globula mul i alen sys ems a e wa e soluble and show no
cy o oxici y in cell lines allowing he s udy o hei po en ial biological unc ion in p e en ing
i al in ec ion. All mul i alen compounds we e checked o he possibili y o blocking DC-SIGN
ecep o in 6 independen expe imen s. The esul s o blocking DC-SIGN ecep o by di e en
compounds we e shown as a unc ion o concen a ion. The 50% o inhibi ion o he in ec ion
was calcula ed wi h he 95% con idence in e al (CI). As a con ol, in ec ion wi h DC-SIGN-
independen VSV glycop o ein-pseudo yped len i i al pa icles29 was pe o med in he same
condi ions.
The esul s ob ained in he in ec ion expe imen e ealed he dependence o he inhibi ion
e ec on mannoses. Compound 17b displaying 120 galac oses, as expec ed, was no able o
inhibi he in ec ion p ocess media ed by DC-SIGN. Compounds wi h 120 mannose-based
esidues (17a and 17c) showed e y s ong an i i al ac i i y a picomola o nanomola
concen a ions. Compound 17a could e ec i ely block Ebola i us in ec ion a low nanomola
concen a ions wi h an IC50 o 20.37 nM (95%CI = 14.63 – 28.37 nM). Compound 17c was
almos one o de o magni ude mo e po en a inhibi ing he in ec ion p ocess wi h an IC50 o
667 pM (95%CI = 411 pM – 1.08 nM) (Figu e 6).
P e ious inhibi ion s udies using he same in ec ion model and ulle enes displaying up o 36
mannoses show ela i e inhibi o po ency (RIP) alues a leas wo o de s o magni ude
smalle .19 Mo eo e , huge i us-like pa icles (VLP) wi h a adius o 16 nm and up o 1640
mannoses32 we e 18- old less po en han compound 17c desc ibed in his wo k (see Table 1).
These esul s ha e con i med he e iciency o hese sys ems o in e ac wi h DC-SIGN and o
compe e wi h Ebola i us glycop o ein-pseudo yped pa icles du ing hei en y in o a ge
cells.
The cy o oxic e ec o mul i alen glyco ulle enes was e i ied by a cell p oli e a ion assay
using he Cell Ti e 96 AQueous Non-Radioac i e Cell P oli e a ion Assay (P omega). No ably,
he e was no any app eciable cy o oxic e ec o compounds 17a-c a he concen a ion used
in he in ec ion expe imen s (see supplemen a y Figu e 6).
Conclusions
In summa y, we ha e syn hesized gian globula mul i alen glyco ulle enes in which he
cen al C60 co e is co alen ly connec ed o wel e hexakis-adduc s o C60, hus o ming he i s
ideca ulle enes epo ed so a . Each pe iphe al ulle ene is endowed wi h en
monosaccha ides and so a o al o 120 ca bohyd a es deco a e he pe iphe y o each
molecule. This ep esen s he as es dend ime g ow h e e epo ed, a o ding molecula
weigh s as high as 56 KDa.
The syn hesis o non-symme ic hexakis-adduc s (7, 9, 14a,b, 15a,b) as well as he
ideca ulle enes (17a-c) ha e been accomplished e icien ly by using CuAAC click-chemis y
eac ions. In his way, e y sophis ica ed molecula ensembles ha e been p oduced in a
minimum o syn he ic s eps and hei appa en s uc u al complexi y is no a limi a ion o
hei applica ions. Despi e he high molecula weigh s, he new molecules ha e been
cha ac e ized by s anda d spec oscopic echniques (FTIR, 1H NMR, 13C NMR) as well as by DLS,
XPS and TEM. In e es ingly, NMR spec oscopy unambiguously e eals he high deg ee o
symme y (Th) in hexakis-adduc s.
Figu e 5. TEM images o ideca ulle ene 17a. These images show small sphe ical pa icles
wi h a diame e o a ound 4 nm co esponding o a single molecule. a) TEM images o
compound 17a upon deposi ion o a 0.01 mg/mL solu ion in H2O. b) De ail o a pa icle
co esponding appa en ly o one molecule. c) Wid h p o ile o he pa icle shown in b) which
has a diame e o  4 nm in acco dance wi h he DLS da a.
Figu e 6. Biological s udy o ideca ulle enes (17a-c). The g aphic shows he inhibi ion o
in ec ion wi h EBOV o VSV GP-pseudo yped len i i al pa icles o Ju ka DC-SIGN+ cells using
17a (blue), 17b (g een) and 17c ( ed). In he cis-in ec ion expe imen s 2.5x105Ju ka DC-SIGN+
we e challenged wi h 5000 Tissue Cul u e In ec i e Dose (TCID) o ecombinan len i i al
pa icles. Resul s ep esen he mean o 6 independen expe imen s +/- SEM. Compounds 17a
and 17c show a s ong inhibi o y ac i i y when EBOV GP pseudo yped len i i al pa icles a e
used. No inhibi o y ac i i y is de ec ed when VSV GP pseudo yped pa icles (con ol) a e used
as in ec i e agen . Compound 17b endowed wi h galac oses as ca bohyd a e uni s does no
show inhibi o y ac i i y because is no able o block DC-SIGN ecep o .
Table 1. Compa a i e o IC50 alues and ela i e inhibi o y po ency o di e en mannosyla ed
mul i alen compounds in inhibi ion s udies using pseudo yped Ebola i us pa icles. The able
shows he da a o he new compounds 17a and 17c desc ibed in compa ison wi h hose
ca bohyd a e mul i alen sys ems p e iously epo ed by us.

Figu e 1.
Figu e 2.
Figu e 3.
Figu e 4.
Figu e 5.
Figu e 6.
10-1 100101102103104105106107108
-20
0
20
40
60
80
100
120
17c-EBOV
17c-VSV-G
17a-EBOV
17a-VSV
17b-EBOV
17b-VSV
concen a ion (pM)
inhibi ion (%)