Accep ed Manusc ip
In es iga ion o glycosyla ing p ope ies o 1-deoxy-1-e hoxysul onyl-hep -2-
ulopy anosyl de i a i es. Syn hesis o a new sul onic acid mime ic o he sialyl
Lewis X e asaccha ide
Magdolna Csá ás, Gábo Máje , Mihály He czeg, Judi Remenyik, László
Lázá , A ila Mándi, Anikó Bo bás, Sándo An us
PII: S0008-6215(11)00207-2
DOI: 10.1016/j.ca es.2011.04.027
Re e ence: CAR 5759
To appea in: Ca bohyd a e Resea ch
Recei ed Da e: 3 Feb ua y 2011
Re ised Da e: 12 Ap il 2011
Accep ed Da e: 18 Ap il 2011
Please ci e his a icle as: Csá ás, M., Máje , G., He czeg, M., Remenyik, J., Lázá , L., Mándi, A., Bo bás, A., An us,
S., In es iga ion o glycosyla ing p ope ies o 1-deoxy-1-e hoxysul onyl-hep -2-ulopy anosyl de i a i es.
Syn hesis o a new sul onic acid mime ic o he sialyl Lewis X e asaccha ide, Ca bohyd a e Resea ch (2011), doi:
10.1016/j.ca es.2011.04.027
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1
In es iga ion o glycosyla ing p ope ies o 1-deoxy-1-e hoxysul onyl-hep -2-
ulopy anosyl de i a i es. Syn hesis o a new sul onic acid mime ic o he sialyl
Lewis X e asaccha ide
Magdolna Csá ás,a Gábo Máje ,a Mihály He czeg,a Judi Remenyik,b László Lázá ,a A ila
Mándi,c Anikó Bo bása* and Sándo An usa,c
aResea ch G oup o Ca bohyd a es o he Hunga ian Academy o Sciences, PO Box 94,
Deb ecen H-4010, Hunga y
bDepa men o Plan Bio echnology, Cen al o Ag icul u al and Applied Economic Sciences,
PO Box 36, Deb ecen H-4015, Hunga y
cDepa men s o O ganic Chemis y, PO Box 20, Facul y o Science, Uni e si y o Deb ecen,
Deb ecen H-4010, Hunga y
Dedica ed o P o esso And ás Lip ák on he occasion o his 75 h bi hday
Abs ac
Glycosyla ion eac ions o he e hyl hio, b omo, and chlo o de i a i es o 1-deoxy-1-
e hoxysul onyl-hep -2-ulopy anose we e s udied applying di e en accep o s unde di e en
condi ions. Elimina ion side- eac ions a o ding exo- and endoglycals occu ed in all cases,
howe e , wi h di e en p opo ions. Glycosyl chlo ide dono was applied o glycosyla e a
isaccha ide accep o ob aining a new sul onic acid mime ic o he sialyl Lewis X
e asaccha ide in high yield.
Keywo ds
2
Ca bohyd a e sul onic acid; Glycosyla ion; Ke opy anosyl glycosides; Elimina ion; Sialyl
Lewis X mime ic
Co esponding au ho . Tel.: +36 52512900/22462; ax: +36 52512900/22342.
E-mail add ess: [email p o ec ed].hu (A. Bo bás).
1. In oduc ion
Selec ins a e ca bohyd a e-binding ansmemb ane glycop o eins, and hei ole is o media e
he i s s eps o he ec ui men o leukocy es om he blood s eam in a se ies o no mal and
pa hologic si ua ions.1 Con ol o hese p ocesses by inhibi ing he adhesion be ween he
calcium-dependen lec in domain o selec ins and hei ca bohyd a e ligands has been
conside ed as a new an i-in lamma o y and an i-me as a ic s a egy.2 Sialyl Lewis X (sLex)
e asaccha ide, one o he majo na u al ligands o selec ins se es as a lead-s uc u e o he
design o selec in an agonis s (Figu e 1).3 The nega i ely cha ged ca boxyla e o sialic acid is
c ucial in binding o ansmemb ane p o eins, bu i can be eplaced by o he cha ged
moie ies. Analogues con aining e.g. sul a e es e ,4 phospha e es e 5 o cyclohexyl lac ic acid6
ins ead o sialic acid p o ed o be ac i e as selec in an agonis s.
Ea lie , we desc ibed he syn hesis o sul onic acid analogues o he sLe X e asaccha ide in
which he sialic acid is eplaced by an anome ic sul onome hyl- ype suga moie y.7,8 To
ins all he sul onome hyl de i a i e o oligosaccha ides e hyl 3,4,5,7- e a-O-benzyl-1-deoxy-
1-e hoxysul onyl-2- hio--D-gluco-hep -2-ulopy anoside was used as glycosyl dono .
Howe e , he yields o glycosyla ion eac ions we e mode a e since a conside able amoun o
exo-glycal was always o med ia elimina ion side- eac ion.
Recen ly, wo isos e ic sul ona e mime ics o mannose-6-O-phospha e p epa ed by
Wadswo h-Ho ne -Emmons ole ina ion ha e been published.9 Bo h conjuga ed and
3
unconjuga ed mannose-sul ona es displayed be e binding a ini y o ca ion-independen
mannose-6-phospha e ecep o and possessed g ea e s abili y in human se um han mannose-
6-phospha e.
We ha e ound ha sul ona e analogues o id apa inux, a syn he ic non-glycosaminoglycan
an icoagulan pen asaccha ide inhibi ed e icien ly he blood coagula ion ac o Xa in an in
i o assay.10
The expe imen al e idence o biological ac i i y o ca bohyd a e sul onic acids inspi ed us o
e isi he syn hesis o sul onic acid mime ic o sialic acid-con aining oligosaccha ides using
anome ic sul onome hyl- ype dono s.
He e, we epo glycosyla ion eac ions o e hyl hio, chlo o and b omo de i a i es o 3,4,5,7-
e a-O-benzyl-1-deoxy-1-e hoxysul onyl--D-gluco-hep -2-ulopy anose applying di e en
accep o s and condi ions. E icien syn hesis o a new sul onic acid e asaccha ide analogue
o he sialyl Lewis X is also discussed.
O
OH
OH
OH
O
AcHN
HO
COO
-
O
HO OH OH
O
O
O
NHAc
OH
O
H
O
H3COH
OH
O
H
Figu e 1. S uc u e o he sialyl Lewis X e asaccha ide
2. Resul s and discussion
2.1. Glycosyla ion s udy
Thioglycoside 2,7,8 and he app op ia e b omo (3) and chlo o (4) de i a i es we e planned o
be u ilized as glycosyl dono s. Compound 2 was ea ed wi h b omine o p oduce 3; he
eac ion wen o comple ion in en minu es. The chlo osuga 4 was p epa ed om he
hemike al 1 using hionyl chlo ide and py idine as eagen s (Scheme 1).
4
O
BnO
OBn
BnO
BnO
SO3E
OH
O
BnO
O
B
n
BnO
BnO
SO3E
R
14
O
BnO
OBn
BnO
BnO
SO3E
Cl
2 R= SE
3 R= B
iiii
ii
Scheme 1. Reagen s and condi ions: i) E SH, BF3E 2O, abs CH2Cl2, 0 °C, 93% ii) B 2, abs
DCM, 0 °C, 10 min, iii) SOCl2, py, abs CH2Cl2, , 10 min, 85%.
The ob ained 2-4 we e used as dono s o glycosyla e me hanol (Scheme 2), accep o 911
con aining ee hyd oxyl g oup a p ima y posi ion (Scheme 3), and accep o s 1211 and 1312
wi h seconda y alcoholic unc ions (Scheme 4), espec i ely. NIS-T OH p omo e sys em was
applied a di e en empe a u es o ac i a e he hioglycoside, and MeOT was also used as a
p omo e o glycosyla ion o 12 and 13.13 Halide dono s, p epa ed eshly be o e
glycosyla ions and used wi hou pu i ica ion, we e ac i a ed wi h ei he AgOT o Hg(CN)2,
and chlo o suga was p omo ed also wi h a mix u e o Hg(CN)2 and HgB 2. Rela i e
p opo ions o he p oduc s we e de e mined om he c ude eac ion mix u e by HPLC
measu emen s.
7
8
O
BnO
BnO
BnO
OCH3
O
B
n
SO3E O
BnO
BnO
BnO OCH3
OBn
SO3E
+
O
OBn
OBn
BnO
BnO SO3E O
OBn
OBn
BnO
BnO SO3E
+
5 6
i
2, 3 o 4
Scheme 2. Reagen s and condi ions: i) MeOH (15 equi ), 3  MS, o sol en s and p omo e s
see Table 1.

5
The glycosyla ion esul s using me hanol as an accep o (Scheme 2) a e summa ized in Table
1. Each eac ion a o ded an anome ic mix u e o he me hyl glycosides 5 and 6.13 Using he
hioglycoside dono 2 he s e eoselec i i y was poo in a ou o ei he he α- o he β-anome
depending on he eac ion empe a u e. Reac ions applying halide dono s 3 and 4 p o ided he
α-isome 5 as he majo p oduc wi h high s e eoselec i i y. F om he chlo o and b omo-
suga s he known elimina ion side-p oduc 77,8 was o med in all cases, and upon AgOT
p omo ion he endoglycal 8 as ano he side-p oduc was obse ed as well.
Table 1. Glycosyla ion o me hanol wi h 1-deoxy-1-e hloxysul onyl-hep -2-ulopy anosyl
dono s
aAmoun o p omo e s: NIS (1.2 equi ) T OH (0.4 equi ); Hg(CN)2 (1 equi ); AgOT (2
equi ); Hg(CN)2-HgB 2 (1.5 equi -0.5 equi ). Sol en s: CH2Cl2 o en ies 1-3, CH2Cl2-
oluene o en ies 4 and 6, CH2Cl2-CH3CN o en ies 5 and 7.
Glycosyla ion eac ions using accep o 9 possessing a eac i e p ima y hyd oxyl (Scheme 3)
a e summa ized in Table 2. In each case a mix u e o he s e eoisome ic 2,6-linked
disaccha ides 10 and 1111 oge he wi h elimina ion side-p oduc (s) was o med. Reac ions
using hioglycoside ga e he disaccha ides wi h he highes p opo ion and he highes α-
En y Dono P omo e
a
T (°C) Time 5 (%) 6 (%) 7 (%) 8 (%)
1 2 NIS-T OH -60 40 min 43 57 - -
2 2 NIS-T OH 0 40 min 62 38 - -
3 2 NIS-T OH 30 min 69 31 - -
4 3 AgOT -40 o -10 20 min 81.3 17.6 0.7 0.4
5 3 Hg(CN)2 0 20 min 65.2 33.1 1.6 -
6 4 AgOT -40 o -10 5 h 80.7 12.0 5.4 1.9
7 4 Hg(CN)2-HgB 2 0 o 5 h 84.9 9.5 5.6 -
6
selec i i y; by using chlo o- and b omo suga s upon AgOT p omo ion a –15 oC a sligh ly
lowe a e o disaccha ides and lowe s e eoselec i i y we e achie ed. Fo bo h halide dono s
he a e o glycosyla ion dec eased using Hg(II) sal ins ead o Ag(I) sal . Ca ying ou
glycosyla ion wi h dono 4 o low eac i i y using less powe ul ac i a ion he glycoside
o ma ion dec eased d ama ically and elimina ion eac ions became he dominan eac ions
(En y 7).
+ 7 +
8
O
O
BnO
BnO
OCH3
BnO
O
BnO
OH
BnO
BnO
OCH3
OSO3E
O
B
n
BnO
BnO
BnO O
O
OBn
BnO
BnO BnO
E O3S
O
BnO
BnO
OCH3
BnO
+
9
1
0
1
1
2, 3 o 4
Scheme 3. Reagen s and condi ions: see Table 2.
Table 2. Glycosyla ion o 9 wi h 1-deoxy-1-e hoxysul onyl-hep -2-ulopy anosyl dono s
En y Dono P omo e a T (°C) Time 10 (%) 11 (%) 7 (%) 8 (%)
1 2 NIS-T OH -50 1 h 81.6 6.7 11.7 -
2 2 NIS-T OH -5 30 min 74.4 7.7 17.9 0
3 3 AgOT -15 7.5 h 63.6 24.0 12.4b
4 3 Hg(CN)2 0 3 h 55.2 19.4 25.4b
5 4 AgOT -15 8 h 71.5 6.3 19.8 2.4
6 4 Hg(CN)2-HgB 2 0 18 h 55.8 19.6 24.6b
7 4 Hg(CN)2 18 h 11.3 8.5 57.1 23.1
aAmoun o p omo e s: NIS (1.2 equi ) T OH (0.4 equi ); Hg(CN)2 (1 equi ); AgOT (2
equi ); Hg(CN)2-HgB 2 (1.5 equi -0.5 equi ). bRa io o 7 and 8 was no de e mined. Sol en s:
7
CH2Cl2 o en ies 1-2, CH2Cl2- oluene o en ies 3 and 5, CH2Cl2-CH3CN o en ies 4, 6
and 7.
Glycosyla ion eac ions using accep o 12 and 13 (Scheme 4) a e summa ized in Tables 3 and
4. The main ea u es o hese eac ions we e ha elimina ion eac ions gene ally o e ook
glycoside o ma ion, and he α-linked disaccha ides o med exclusi ely. Con igu a ion o he
in e glycosidic linkages o 14 and 15 was de e mined on he basis o he NMR C1–H3 h ee-
bond coupling cons an which is dependen on he dihed al angle in a manne simila o
3JH,H.13
Thioglycoside 2 upon powe ul ac i a ion using NIS-T OH was ine icien o glycosyla ion
o he un eac i e accep o s 12 and 13; eac i i y o he seconda y hyd oxyl g oups was low a
he low empe a u e o ac i a ion (-40 °C), he e o e oxoca benium in e media e o med
apidly om he dono was s abilized ia elimina ion (Table 3: en ies 1, 2 and Table 4: en y
1). Slow ac i a ion p ocess o 2 a oom empe a u e using he mild MeOT as a p omo e
ga e sligh ly be e esul s, due o he inc easing eac i i y o he accep o a highe
empe a u e (Table 3: en y 3 and Table 4: en y 2). Beside exoglycal 7, o ma ion o
endoglycal 8 om hioglycoside was also obse ed.
Glycosyl halides p o ed o be mo e e icien dono s o bo h accep o s 12 and 13 han he
hioglycoside. In e es ingly, he ou come o he eac ions was g ea ly in luenced by he
aglycons o he accep o s (α-me hyl o 12 and β-p-me hoxyphenyl o 13). Fo accep o 12
he mo e eac i e halide dono 3 wi h he less powe ul p omo e Hg(CN)2 wo ked as well
han he less eac i e halide dono 4 wi h he mo e powe ul p omo e AgOT (Table 3:
en ies 5 and 6). Fo accep o 13 chlo osuga 4 upon AgOT p omo ion wo ked much be e
han b omosuga 3 ac i a ed by Hg(CN)2 (Table 4: en ies 5 and 4).
8
O
BnO
O
R2
BnO
O
BnO
BnO
HO
R2
OSO3E
O
B
n
BnO
BnO BnO
OBn
OBn
12 R1 = H, R2 = OCH3 14
13 R1 = OPMP, R2 = H 15
R1
R1+ 7 +
8
2, 3 o 4
Scheme 4. Reagen s and condi ions: see Table 3 and 4.
Table 3. Glycosyla ion o 12 wi h 1-deoxy-1-e hoxysul onyl-hep -2-ulopy anosyl dono s
En y Dono
P omo e a T (°C) Time 14 (%) 7 (%) 8 (%)
1 2 NIS-T OH -40 30 min 17.4 55.1 27.5
2 2 NIS-T OH 0 2 h 0.6 99.4b
3 2 MeOT 1 day 23.2 76.8b
4 3 AgOT 0 1 h 28.6 40.8 30.6
5 3 Hg(CN)2 0 1h 39.6 35.0 25.4
6 4 AgOT 0 1h 40.0 45.2 14.8
7 4 Hg(CN)2-HgB 2 0 4 days 11.2 62.0 26.8
aAmoun o p omo e s: NIS (1.2 equi ) T OH (0.4 equi ); MeOT (6 equi ); AgOT (2
equi ); Hg(CN)2 (1 equi ); Hg(CN)2-HgB 2 (1.5 equi -0.5 equi ). bRa io o 7 and 8 was no
de e mined. Sol en s: CH2Cl2 o en ies 1-3, CH2Cl2- oluene o en ies 4 and 6, CH2Cl2-
CH3CN o en ies 5 and 7.
Table 4. Glycosyla ion o 13 wi h 1-deoxy-1-e hoxysul onyl-hep -2-ulopy anosyl dono s
En y Dono
P omo e a T (°C) Time 15 (%) 7 (%) 8 (%)
1 2 NIS-T OH -40 65 min 12.0 86.1 1.9
15
E 3N, he insoluble ma e ials we e emo ed by il a ion, he il a e was dilu ed wi h CH2Cl2,
washed wi h wa e , il e ed, d ied and concen a ed. The a io o he p oduc s was de e mined
om he c ude p oduc by HPLC.
3.6. Gene al me hod E o p epa a ion o glycosides 5, 6, 10, 11, 14 and 15 wi h halide
dono s 3 and 4
I) To a solu ion o accep o (1.5 equi ) in oluene (2.5 mL) 4 Å molecula sie es and AgOT
(2 equi ) we e added and i was s i ed o 3 h. The eac ion mix u e was cooled o he
app op ia e empe a u e hen he solu ion o dono (1 equi ) in d y CH2Cl2 (2 mL/0.3 mmol)
was added. When TLC showed comple e con e sion o he dono , he insoluble ma e ials
we e emo ed by il a ion, he il a e was dilu ed wi h CH2Cl2, washed wi h 10% Na2S2O3-
solu ion and wa e , il e ed, d ied and concen a ed. The a io o he p oduc s was de e mined
om he c ude p oduc by HPLC.
II) A mix u e o accep o (1.5 equi ) in d y CH3CN (2.5 mL), powde ed 4 Å molecula sie es
and (HgCN)2 (1 equi ) o Hg(CN)2 (1.5 equi ) and HgB 2 (0.5 equi ) was s i ed o 3 h. The
eac ion mix u e was cooled o 0°C hen he solu ion o dono (1 equi ) in d y CH2Cl2 (2
mL/0.3 mmol) was added. When TLC showed comple e con e sion o he dono , he
insoluble ma e ials we e emo ed by il a ion, he il a e was dilu ed wi h CH2Cl2, washed
wi h 10% Na2S2O3-solu ion and wa e , il e ed, d ied and concen a ed. The a io o he
p oduc s was de e mined om he c ude p oduc by HPLC.
3.7. 3,4,5,7-Te a-O-benzyl-1-deoxy-1-e hoxysul onyl-α
αα
α-D-gluco-hep -2-ulopy anosyl-
b omide (3)
To a s i ed solu ion o 2 (212 mg, 0.30 mmol) in abs dichlo ome hane (2 mL), 15 L (0.30

16
mmol, 1.0 equi ) b omine was added and i was cooled o 0 oC. The eac ion mix u e was
concen a ed a e 10 min and he esidue was coe apo a ed 3 imes wi h oluene. The
ob ained sy upy c ude p oduc (218 mg) was used o u he eac ion wi hou pu i ica ion. R
0.75 (98:2 CH2Cl2-ace one).
3.8. 3,4,5,7-Te a-O-benzyl-1,2-dideoxy-1-e hoxysul onyl-α
αα
α-D-gluco-hep -2-ulopy anosyl-
chlo ide (4)
Compound 1 (700 mg, 0.99 mmol) was dissol ed in 5 mL abs dichlo ome hane and 55 L
(0.99 mmol, 1.0 equi ) py idine and 85 L (1.09 mmol, 1.1 equi ) SOCl2 we e added a oom
empe a u e. A e 10 min he eac ion mix u e was dilu ed wi h CH2Cl2, washed wice wi h
1M aq HCl, 1M aq NaHCO3 and wa e , hen i was d ied, il e ed, e apo a ed and pu i ied by
column ch oma og aphy, o gi e 4 (625 mg, 85%). Fo glycosyla ion, he sy upy c ude
p oduc was used wi hou pu i ica ion. R 0.81 (n-hexane-E OAc 7:3); 1H NMR (200 MHz,
CDCl3):
δ
7.38-7.12 (m, 20H, Ph), 5.04 (m, 7H, J 11.9 Hz, CH2Ph), 4.59 (d, 1H, J 9.2 Hz),
4.50 (d, 1H, J 12.1 Hz, CH2Ph), 4.28-4.13 (m, 3H, SO3CH2CH3), 4.13-4.06 (m, 1H), 3.80 (d,
1H, J 15.1 Hz, H-1a), 3.82-3.72 (m, 3H), 3.69 (d, 1H, J 15.1 Hz, H-1b), 1.22 ( , 3H, J 7.1 Hz,
SO3CH2CH3); 13C NMR (50 MHz, CDCl3)
δ
138.1, 137.9 (4C, Cq, Ph), 128.5-125.6 (20C,
Ph), 104.3 (C-2), 83.6, 79.8, 76.7, (C-3, C-4, C-5), 75.6, 75.2, 75.1, 73.3 (4C, CH2Ph), 68.0
(2C, C-7, SO3CH2CH3), 57.9 (C-1), 15.0 (SO3CH2CH3); MALDI-TOF m/z Calcd o
C37H41ClNaO8S+ [M+Na]+: 703.21, Found: 703.21.
3.9. 2,6-Anhyd o-3,4,5,7- e a-O-benzyl-1-e hoxysul onyl-D-gluco-hep -2-eni ol (8)
Isola ed as a colou less sy up (14%) om he eac ion o 4 and 9 acco ding o gene al me hod
E, II, using Hg(CN)2 as a p omo e . [α]D -4.3 (c 0.07, CHCl3); R 0.60 (99:1 CH2Cl2-ace one);
17
1H NMR (400 MHz, CDCl3):
δ
7.34-7.25 (m, 20H, Ph), 4.85-4.48 (m, 8H, CH2Ph), 4.39 (d,
1H, J 4.2 Hz, H-4), 4.27-4.21 (m, 3H, H-6, SO3CH2CH3), 4.06-3.97 (m, 3H, H-5, C-1-CH2),
3.83-3.75 (m, 2H, C-7a,b), 1.25 ( , 3H, J 7.1 Hz, SO3CH2CH3); 13C NMR (100 MHz, CDCl3):
δ
137.9, 137.7, 137.6, 136.9 (4C, Cq, Ph), 136.1, 135.7 (C-2 and C-3), 128.4-127.6 (20C, Ph),
76.5 (C-6), 73.5 (C-4), 72.6 (C-5), 73.6, 73.4, 72.5, 70.4 (4C, 4 x CH2Ph), 67.9
(SO3CH2CH3), 67.8 (C-7), 49.7 (C-1), 15.0 (SO3CH2CH3); Anal. Calcd o : C37H40O8S
(644.77): C, 68.92; H, 6.25; S, 4.97. Found: C, 68.79; H, 6.29; S, 5.00.
3.10. Me hyl 2,3,6- i-O-benzyl-4-O-(3,4,5,7- e a-O-benzyl-1-deoxy-1-e hoxysul onyl-α
αα
α-
D-gluco-hep -2-ulopy anosyl)-β
ββ
β-D-glucopy anoside (14)
Compound 14 was p epa ed om dono 2 and accep o 12 by me hod C and dono 3 o 4 and
accep o 12 by me hod E (see Table 3). [α]D +33.1 (c 0.19, CHCl3); R 0.69 (98:2 CH2Cl2-
ace one); 1H NMR (500 MHz, CDCl3)
δ
7.33-7.14 (m, 35H, Ph), 4.95-4.84 (m, 5H, CH2Ph),
4.77 (d, 1H, J 11.1 Hz, CH2Ph), 4.66 (d, 1H, J 11.2 Hz, CH2Ph), 4.60-4.57 (m, 5H, H-1,
CH2Ph), 4.51-4.42 (m, 3H, CH2Ph), 4.21-4.14 (m, 4H, H-1’a, H-5’, H-6’, H-7’a), 4.05 ( , 1H,
J 9.0 Hz, H-4), 4.00 (dd, 1H, J 10.0 Hz, J 5.0 Hz, H-3’), 3.91-3.89 (m, 1H, H-3), 3.86 (q, 2H,
J 7.0 Hz, SO3CH2CH3), 3.73-3.70 (m, 3H, H-1’b, H-5, H-6a), 3.67-3.62 (m, 2H, H-4’, H-6b),
3.55 (d, 1H, J 11.3 Hz, H-7’b), 3.45 (dd, 1H, J1,2 3.3 Hz, J2,3 9.5 Hz, H-2), 3.36 (s, 3H,
OCH3), 1.08 ( , 3H, J 7.05 Hz, SO3CH2CH3 ); 13C NMR (125 MHz, CDCl3)
δ
139.1-137.8
(7C, Cq, Ph), 128.6-126.8 (35C, Ph), 99.9 (C-2’), 97.3 (C-1), 82.4 (C-5’), 81.0 (C-3), 80.4 (C-
6’), 79.4 (C-2), 78.5 (C-4’), 73.2 (C-4), 73.0 (C-3’), 70.3 (C-5), 75.4, 75.3, 75.2, 73.3, 73.2,
73.1 (7C, CH2Ph), 69.5 (C-7’), 68.9 (C-6), 66.7 (SO3CH2CH3), 55.4 (OCH3), 51.5 (C-1’,
3JC1’,H3’ ≤ 1 Hz), 14.8 (SO3CH2CH3); Anal. Calcd o : C65H72O14S (1109.32): C, 70.38; H,
6.54; S, 2.89. Found: C, 69.96; H, 6.33; S, 2.76.
18
3.11. 4-Me hoxyphenyl 2,3,6- i-O-benzyl-4-O-(3,4,5,7- e a-O-benzyl-1-deoxy-1-
e hoxysul onyl-α
αα
α-D-gluco-hep -2-ulopy anosyl)-β
ββ
β-D-glucopy anoside (15)
Compound 15 was p epa ed om dono 2 and accep o 13 by me hod C and om dono 3 o
4 and accep o 13 by me hod E (see Table 4). [α]D +9.8 (c 0.17, CHCl3); R 0.85 (97:3
CH2Cl2-ace one); 1H NMR (500 MHz, CDCl3)
δ
7.34-7.16 (m, 35H, Ph), 6.98, 6.79 (2 x d,
4H, PMP), 5. 07 (d, 1H, J 6.3 Hz), 4.98-4.41 (m, 14H, CH2Ph), 4.23 (d, 1H, J 9.6 Hz), 4.17-
4.11 (m, 3H), 3.96-3.89 (m, 5H), 3.75 (s, 3H, OCH3), 3.71-3.59 (m, 7H), 1.10 ( , 3H, J 7.1
Hz, SO3CH2CH3); 13C NMR (125 MHz, CDCl3)
δ
155.1, 151.2 (2C, Cq, PMP), 138.4-138.0
(7C, Cq, Ph), 128.3-127.1 (35C, Ph), 118.1, 114.5 (4C, PMP), 102.0 (C-1), 100.1 (C-2’), 82.5,
80.5, 80.1, 78.3, 76.6, 73.3, 73.2 (skele on ca bons) 75.3, 75.2, 75.0, 74.6, 74.1, 73.4, 73.3
(6C, CH2Ph), 70.1, 68.9 (C-6 and C-7’), 67.0 (SO3CH2CH3), 55.5 (OCH3), 51.8 (C-1’, 3JC1’,H3’
≤ 1 Hz), 14.9 (SO3CH2CH3); Anal. Calcd o C71H76O15S (1200.49): C, 70.98; H, 6.38; S,
2.67. Found: C, 71.04; H, 6.43; S, 2.59.
3.12. 4-Me hoxyphenyl 3,4-O-isop opylidene-2,6-di-O-benzoyl-β
ββ
β-D-galac opy anosyl-
(14)-2,6-di-O-benzoyl-β
ββ
β-D-glucopy anoside (17)
A s i ed solu ion o 16 (970 mg, 2 mmol) in 15 mL o d y py idin and 20 mL o d y oluene
was cooled o 0 °C and BzCl (1.85 ml, 16 mmol, 8 equi ) was added d opwise. A e 90 min
MeOH was added and he eac ion mix u e was concen a ed. I was hen dilu ed wi h E OAc,
washed wi h sa d aq NaHCO3 and wa e , d ied, il e ed and e apo a ed. The c ude p oduc
was ec ys allized om E OAc-n-hexane o yield 17 (1.25 g, 69%). Mp 225-227°C, []D +7.6
(c 0.54, CHCl3); R 0.73 (95:5 CH2Cl2-ace one); 1H NMR (200 MHz, DMSO-d6)
δ
8.09-7.29
19
(m, 20H, Ph), 6.82, 6.59 (2 x d, 4H, PMP), 5.46 ( , 1H, J 8.5 Hz),5.38 ( , 1H, J 7.6 Hz), 4.7 (d,
1H, J1,2 8.0 Hz,), 4.87 (d, 1H, J 10.8 Hz), 4.69-4.66 (m, 2H), 4.47-4.28 (m, 6H), 4.06 ( , 1H, J
9.2 Hz), 3.84-3.75 (m, 2H), 3.65 (s, 3H, OCH3), 1.64, 1.35 (2 x s, 6H, C(CH3)2); 13C NMR
(50 MHz, DMSO)
δ
166.5, 165.4, 165.2 (4C, CO), 155.4, 151.0 (2C, Cq, PMP), 133.3-128.4
(24C, Ph), 118.7, 114.2 (4C, PMP), 111.2 (C(CH3)2), 101.4 (C-1), 100.5 (C-1’), 82.4, 76.9,
73.5, 73.4, 72.9, 72.7, 72.1, 72.0 (skele on ca bons), 63.6, 62.7 (C-6 and C-6’), 55.4 (OCH3),
27.5, 26.2 (C(CH3)2); Anal. Calcd o C50H48O16 (904.29): C, 66.36; H, 5.35. Found: C, 66.37;
H, 5.34.
3.13. 4-Me hoxyphenyl 2,6-di-O-benzoyl-3,4-O-isop opylidene-β
ββ
β-D-galac opy anosyl-
(14)-2,6-di-O-benzoyl-3-O-(2,3,4- i-O-benzyl-α
αα
α-L- ucopy anosyl)-β
ββ
β-D-
glucopy anoside (19)
To a s i ed solu ion o accep o 17 (1.1 g, 1.22 mmol) and dono 18 (0.875 g, 1.83 mmol) in
d y CH2Cl2 (20 mL) 4 Å molecula sie es was added. A e s i ing o 1 h he mix u e was
cooled o -45 °C and NIS (36 mg, 1.3 equi ) and TMSOT (66 µL, 0.3 equi ) dissol ed in
CH2Cl2:THF (1:1, 5 mL) we e added. A e 1 h he eac ion was quenched wi h py idine (0.5
mL), dilu ed wi h CH2Cl2 and il e ed h ough Celi e. The il a e was washed wi h 10% aq
Na2S2O3 and wa e , d ied and concen a ed. The c ude p oduc was pu i ied by column
ch oma og aphy (99:1 CH2Cl2-ace one) o yield 19 (1.51 g, 94%) as a colou less sy up. []D
+13.56 (c 0.649, CHCl3); R 0.42 (n-hexane-E OAc 7:3); 1H NMR (500 MHz, CDCl3)
δ
8.12-
6.90 (m, 35H, Ph), 6.65, 6.47 (2 x d, 4H, PMP), 5.58 ( , 1H, J 8.8 Hz), 5.38 (d, 1H, J1”,2” 3.5
Hz, H-1”), 5.19 ( , 1H, J 8.0 Hz), 4.90-4.69 (m, 6H), 4.61-4.44 (m, 4H), 4.33-4.18 (m, 6H),
4.11-4.02 (m, 2H), 3.90-3.89 (m, 2H), 3.73 (s, 1H), 3.58-3.54 (m, 1H), 3.53 (s, 3H, OCH3),
1.43, 1.24 (2 x s, 6H, C(CH3)2), 1.20 (d, 3H, H-6”); 13C NMR (125 MHz, CDCl3)
δ
166.3,
20
165.8, 164.6, 164.4 (4C, CO), 155.4, 151.0 (2C, Cq, PMP), 139.0. 138.5, 138.0 (3C, Cq, Ph),
133.3-126.7 (39C, Ph), 118.6, 114.2 (4C, PMP), 110.7 (C(CH3)2), 100.8 (C-1), 100.3 (C-1’),
97.3 (C-1”), 79.0, 78.1, 75.5, 73.9, 73.4, 73.2, 73.1, 71.3, 66.4 (skele on ca bons), 74.5, 72.5
(3C, CH2Ph), 62.5 (C-6 and C-6’), 55.3 (OCH3), 27.6, 26.2 (C(CH3)2), 16.8 (C-6”). Anal.
Calcd o C77H76O20 (1320.49): C, 69.99; H, 5.80. Found: C, 66.97; H, 5.78.
3.14. 4-Me hoxyphenyl 2,6-di-O-benzoyl-β
ββ
β-D-galac opy anosyl-(14)-2,6-di-O-benzoyl-
3-O-(2,3,4- i-O-benzyl-α
αα
α-L- ucopy anosyl)-β
ββ
β-D-glucopy anoside (20)
A solu ion o 19 (1.3 g, 0.19 mmol) in MeOH (25 mL) was ea ed wi h 2.5 mL 1M aq HCl a
50 °C o 3 h. The mix u e was concen a ed, dilu ed wi h CH2Cl2, washed wi h sa d aq
NaHCO3 and wa e , d ied, il e ed and e apo a ed. The c ude p oduc was pu i ied by column
ch oma og aphy (9:1 CH2Cl2-E OAc) o yield 20 (1.1 g, 89%) as a colou less sy up. []D -
11.6 (c 0.96, CHCl3); R 0.52 (9:1 CH2Cl2-E OAc); 1H NMR (200 MHz, CDCl3)
δ
8.12-6.98
(m, 35H, Ph), 6.81-6.52 (m, 4H, PMP), 5.69 ( , 1H, J 9.0 Hz, H-2), 5.40 (d, 1H, J1”,2” 3.5 Hz,
H-1”), 5.25 ( , 1H, J 9.0 Hz, H-2’), 4.89-4.53 (m, 11H), 4.39-4.07 (m, 6H), 3.99-3.53 (m, 7H),
3.63 (s, 3H, OCH3), 1.32 (d, 3H, H-6”); 13C NMR (50 MHz, CDCl3)
δ
166.5, 166.2, 166.0,
164.5 (4C, CO), 155.3, 151.1 (2C, Cq, PMP), 139.1-126.7 (35C, Ph), 118.6, 114.2 (4C, PMP),
100.9 (C-1), 100.1 (C-1’), 97.6 (C-1”), 78.9, 78.3, 75.4, 74.4, 73.5, 71.8, 67.7, 66.5 (skele on
ca bons), 75.1, 72.6 (3C, CH2Ph), 62.9, 61.7 (C-6 and C-6’), 55.4 (OCH3), 16.5 (C-6”). Anal.
Calcd o C74H72O20 (1280.46): C, 69.36; H, 5.66. Found: C, 66.34; H, 5.65.
3.15. 4-Me hoxyphenyl 3,4,5,7- e a-O-benzyl-1-deoxy-1-e hoxysul onyl-α
αα
α-D-gluco-hep -
2-ulopy anosyl-(13)-2,6-di-O-benzoyl-β
ββ
β-D-galac opy anosyl-(14)-2,6-di-O-benzoyl-3-
O-(2,3,4- i-O-benzyl-α
αα
α-L- ucopy anosyl)-β
ββ
β-D-glucopy anoside (21)

21
To a s i ed solu ion o accep o 20 (750 mg, 0.57 mmol) and dono 4 (267 mg, 0.38 mmol) in
d y CH2Cl2 (10 mL) 4 Å molecula sie es was added. A e s i ing o 1 h he mix u e was
cooled o 0 °C and AgOT (74 mg, 2 equi ) dissol ed in oluene (2 mL) was added. The
mix u e was kep a 0 °C o e nigh . Insoluble ma e ials we e emo ed by il a ion, he il a e
was dilu ed wi h CH2Cl2, washed wi h 10% aq Na2S2O3 and wa e , d ied, il e ed and
concen a ed. The c ude p oduc was pu i ied by column ch oma og aphy (6:4 n-hexane-
E OAc) o yield 21 (710 mg, 59%) as a colou less sy up.
Compound 21 was also p epa ed om accep o 20 (250 mg, 0.19 mmol, 1.5 equi ) and dono
2 (92 mg, 0.13 mmol, 1 equi ) in d y CH2Cl2 (10 mL) by me hod D using MeOT (86 µL,
0.78 mmol, 6 equi ). The c ude p oduc was pu i ied by column ch oma og aphy (9:1
oluene-ace one) o yield 21 (82 mg, 33%) as a colou less sy up.
Compound 21 was also p epa ed om accep o 20 (250 mg, 0.19 mmol, 1.5 equi ) and dono
2 (92 mg, 0.13 mmol, 1 equi ) in d y CH2Cl2 (10 mL) by me hod C using NIS (35 mg, 1.2
equi ) and T OH (4.7 µL, 0.05 mmol, 0.4 equi ). The c ude p oduc was pu i ied by column
ch oma og aphy (9:1 oluene-ace one) o yield 21 (25 mg, 10%) as a colou less sy up. []D
+24.4 (c 0.38 CHCl3); R 0.37 (9:1 oluene-ace one); 1H NMR (400 MHz, CDCl3)
δ
8.15-7.00
(m, 55H, Ph), 6.74, 6.55 (2 x d, 4H, PMP), 5.61 ( , 1H, J2,3 8.2 Hz, H-1), 5.47 ( , 1H, J2’,3’ 8.9
Hz, H-1’), 5.41 (d, 1H, J1”,2” 3.5 Hz, H-1”), 4.98-4.17 (m, 28H, 7 x CH2Ph, H-5”, H-1, H-1’,
H-6a,b, H-6’a,b, H-4”, H-7’’’a,b, H-6’’’, H-3, H-4’’’, H-3”), 4.11-3.98 (m, 4H, H-4, H-3’,
SO3CH2CH3), 3.95-3.84 (m, 2H, H-2”, H-5’’’), 3.78-3.57 (m, 4H, H-4’, H-3’’’, H-5, H-5’),
3.64 (s, 3H, OCH3), 3.28 (s, 1H, H-4’-OH), 3.15-3.04 (m, 2H, H-1’’’a,b), 1.40 (d, 3H, J5”,6”
6.5 Hz, H-6”), 1.10 ( , 3H, J 7.0 Hz, SO3CH2CH3); 13C NMR (100 MHz, CDCl3)
δ
166.3,
22
165.9, 164.4 (4C, CO), 155.4, 151.1 (2C, Cq, PMP), 139.3-137.3 (7C, Cq, Ph), 129.8-126.7
(55C, Ph), 118.7, 114.2 (4C, PMP), 100.1, 100.4, 97.7 (C-1, C-1’, C-1”), 99.8 (C-2’”), 82.8,
82.4, 80.9, 78.9, 78.8, 77.9, 75.6, 74.5, 74.4, 73.5, 72.8, 71.9, 71.5, 70.6, 67.8, 66.6, (skele on
ca bons), 75.2, 75.1, 74.9, 73.4, 73.1, 72.6, 72.3 (7C, CH2Ph), 68.4, 68.1 (C-6’’’,
SO3CH2CH3), 62.6, 62.2 (C-6, C-6’), 55.4 (OCH3), 53.3 (C-1’’’, 3JC1’’’,H3’’’ ≤ 1 Hz), 16.5 (C-
6”), 15.0 (SO3CH2CH3). MALDI-TOF m/z Calcd o C111H112NaO28S [M+Na]+: 1947.70,
Found: 1948.25. Anal. Calcd o C111H112O28S (1924.71): C, 69.22; H, 5.86; S, 1.66. Found:
C, 69.21; H, 5.84; S, 1.64.
Acknowledgemen s:
This wo k was suppo ed by he TÁMOP 4.2.1/B-09/1/KONV-2010-0007 p ojec . The p ojec
was co- inanced by he Eu opean Union and he Eu opean Social Fund. Financial suppo o
he Hunga ian Resea ch Fund (K 62802) is also acknowledged.
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31
Table 1. Glycosyla ion o me hanol wi h 1-deoxy-1-e hloxysul onyl-hep -2-ulopy anosyl
dono s
aAmoun o p omo e s: NIS (1.2 equi ) T OH (0.4 equi ); Hg(CN)2 (1 equi ); AgOT (2
equi ); Hg(CN)2 (1.5 equi ) HgB 2 (0.5 equi ).
Dono P omo e
a
T (°C) Time 5 (%) 6 (%) 7 (%) 8 (%)
2 NIS-T OH -60 40 min 43 57 - -
2 NIS-T OH 0 40 min 62 38 - -
2 NIS-T OH 30 min 69 31 - -
3 AgOT -40 o -10 20 min 81.3 17.6 0.7 0.4
3 Hg(CN)2 0 20 min 65.2 33.1 1.6 -
4 AgOT -40 o -10 5 h 80.7 12.0 5.4 1.9
4 Hg(CN)2-HgB 2 0 o 5 h 84.9 9.5 5.6 -

32
Table 2. Glycosyla ion o 9 wi h 1-deoxy-1-e hoxysul onyl-hep -2-ulopy anosyl dono s
Dono P omo e a T (°C) Time 10 (%) 11 (%) 7 (%) 8 (%)
2 NIS-T OH -50 1 h 81.6 6.7 11.7 -
2 NIS-T OH -5 30 min 74.4 7.7
3 AgOT -15 7.5 h 63.6 24.0 12.4b
3 Hg(CN)2 0 3 h 55.2 19.4 25.4b
4 AgOT -15 8 h 71.5 6.3 19.8 2.4
4 Hg(CN)2-HgB 2 0 18 h 55.8 19.6 24.6b
4 Hg(CN)2 18 h 11.3 8.5 57.1 23.1
aAmoun o p omo e s: NIS (1.2 equi ) T OH (0.4 equi ); Hg(CN)2 (1 equi ); AgOT (2
equi ); Hg(CN)2 (1.5 equi ) HgB 2 (0.5 equi ); bRa io o 7 and 8 was no de e mined.
33
Table 3. Glycosyla ion o 12 wi h 1-deoxy-1-e hoxysul onyl-hep -2-ulopy anosyl dono s
Dono P omo e a T (°C) Time 14 (%) 7 (%) 8 (%)
2 NIS-T OH -40 30 min 17.4 55.1 27.5
2 NIS-T OH 0 2 h 0.6 99.4b
2 MeOT 1 day 23.2 76.8b
3 AgOT 0 1 h 28.6 40.8 30.6
3 Hg(CN)2 0 1h 39.6 35.0 25.4
4 AgOT 0 1h 40.0 45.2 14.8
4 Hg(CN)2-HgB 2 0 4 days 11.2 62.0 26.8
aAmoun o p omo e s: NIS (1.2 equi ) T OH (0.4 equi ); MeOT (6 equi ); AgOT (2
equi ); Hg(CN)2 (1 equi ); Hg(CN)2 (1.5 equi ) HgB 2 (0.5 equi ). bRa io o 7 and 8 was no
de e mined.
34
Table 4. Glycosyla ion o 13 wi h 1-deoxy-1-e hoxysul onyl-hep -2-ulopy anosyl dono s
Dono P omo e a T (°C) Time 15 (%) 7 (%) 8 (%)
2 NIS-T OH -40 65 min 12.0 86.1 1.9
2 MeOT RT 1 day 18.5 81.5b
3 AgOT 0 2 h 13.5 30.5 56.0
3 Hg(CN)2 0 2.5 h 16.2 29.3 54.5
4 AgOT 0 2 days 63.4 29.1 7.5
4 Hg(CN)2-HgB 2 0 5 days 16.3 50.7 33.0
aAmoun o p omo e s: NIS (1.2 equi ) T OH (0.4 equi ); MeOT (6 equi ); AgOT (2
equi ); Hg(CN)2 (1 equi ); Hg(CN)2 (1.5 equi ) HgB 2 (0.5 equi ). bRa io o 7 and 8 was no
de e mined.
35
In es iga ion o glycosyla ing p ope ies o 1-deoxy-1-e hoxysul onyl-hep -2-ulopy anosyl de i a i es.
Syn hesis o a new sul onic acid mime ic o he sialyl Lewis X e asaccha ide
Magdolna Csá ás, Gábo Máje , Mihály He czeg, Judi Remenyik, László Lázá , A ila Mándi, Anikó Bo bás* and Sándo An us
Resea ch G oup o Ca bohyd a es o he Hunga ian Academy o Sciences, H-4010 Deb ecen, P.O. Box 94, Hunga y