Synthesis of cis-(1 → 3)-glycosides of allyl 2-acetamido-4,6-O- benzylidene-2-deoxy-α-D-glucopyranoside

Article abstract of DOI:10.1016/j.carres.2004.01.014

Syntheses of allyl 2,3,4-tri-O-benzyl-α-D-gluco- and D-galactopyranosyluronate-(1→3)-2-acetamido-4,6-O-benzylidene-2-deoxy- α-D-glucopyranoside via oxidation of the hydroxymethyl group of allyl 2,3,4-tri-O-benzyl-α-D-gluco- and D-galactopyranosyl-(1→3)-2-

Full text of DOI:10.1016/j.carres.2004.01.014

Carbohydrate
RESEARCH
Carbohydrate Research 339 (2004) 1293–1300
Synthesis of cis-(1 fi 3)-glycosides of allyl 2-acetamido-4,6-
O-benzylidene-2-deoxy-a- -glucopyranoside
D
*
ꢀ
Janusz Madaj, Magdalena Jankowska and Andrzej Wisniewski
Department of Chemistry, Sugar Chemistry Group, University of Gdaꢀnsk, ul. Sobieskiego 18, PL-80-952 Gdaꢀnsk, Poland
Received 27 November2003; accepted 25 January 2004
Available online 24 March 2004
Abstract—Syntheses of allyl 2,3,4-tri-O-benzyl-a-
D
-gluco- and
D
-galactopyranosyluronate-(1 fi 3)-2-acetamido-4,6-O-benzylidene-
-gluco- and -galactopyr-
D-glucopyranoside under Jones conditions are described. Structures of the
2-deoxy-a- -glucopyranoside via oxidation of the hydroxymethyl group of allyl 2,3,4-tri-O-benzyl-a-
D
D
D
anosyl-(1 fi 3)-2-acetamido-4,6-O-benzylidene-2-deoxy-a-
1
title compounds were confirmed by H and 13C NMR spectroscopy.
ꢀ 2004 Elsevier Ltd. All rights reserved.
Keywords: Uronic acid; 2-Acetamido-2-deoxy-D
-glucopyranose; Jones oxidation; 1H and 13C NMR spectroscopy
1. Introduction
2. Results and discussion
Uronic acids are of important biological significance and
have attracted the attention of many chemists. Their
glycosides are found in many polysaccharides of mi-
crobes, plants, and animals.1;2 They play important roles
in biological recognition and cellular communication.3
Reaction of the O-allyl group with cysteamine10 forms a
commonly used linkerto BSA ( bovine serum albumin) or
HSA (human serum albumin). Allyl 2-acetamido-4,6-O-
benzylidene-2-deoxy-a-
from allyl 2-acetamido-2-deoxy-a-
(1), was used as a glycosyl acceptor. Direct reaction
between 2 and methyl 2,3,4-tri-O-acetyl-a- -glucopyr-
D
-glucopyranoside (2), prepared
D
-glucopyranoside11
Linked to uronic acids (of the D-gluco-, D-galacto-, or
D
-ido- configurations) they are lipopolysaccharide com-
D
ponents of bacteria cell walls. Glycoproteins and lipo-
polysaccharides with this type of sugar moiety are not
easily available from natural sources because of their
microheterogeneity. In this paper we describe syntheses
of disaccharide fragments of lipopolysaccharides (LPS)
anosyluronate bromide12 (3) led to the intermolecular O-
acetyl group migration.13 This type of side reaction is
well known in the literature14;15 and may be the result of
low reactivity of uronic acid derivatives.16;17 Because the
direct reaction of 2 and 3 was ineffective, we decided to
change the strategy of synthesis. In some cases an
alternative method of preparation of oligosaccharides
containing uronic acid units via oxidation of the hy-
droxymethyl group in appropriately prepared substrate
oligosaccharides, was used.18–20 Based on ourpervious
results,21 we chose the method of Kong and co-work-
ers22 to obtain a free terminal hydroxyl group. The
isolated from Gram-negative bacterial cell walls: a-
GlcpA-(1 fi 3)-
Hafnia alvei PCM 11855 and Proteus mirabilis 010;6 a-
D
-
D
-GlcpNAc from Shigella boydii 54 and
D
-
GalpA-(1 fi 3)-
D
-GlcpNAc from Escherichia coli O657
and Vibrio cholerae O228 and Proteus vulgaris O46.9
reaction of 2 and 2,3,4,6-tetra-O-benzyl-a-
anosyl trichloroacetimidate [5, obtained from 2,3,4,6-
tetra-O-benzyl-a- -glucopyranose (4)] carried out in
D-glucopyr-
* Corresponding author. Fax: +48-58-345-04-72; e-mail: januszm@
chemik.chem.univ.gda.pl
D
0008-6215/$ - see front matter ꢀ 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carres.2004.01.014

1294
J. Madaj et al. / Carbohydrate Research 339 (2004) 1293–1300
DMF gave the expected allyl 2,3,4,6-tetra-O-benzyl-a-
-glucopyranosyl-(1 fi 3)-2-acetamido-4,6-O-benzylidene-
2-deoxy-a- -glucopyranoside (6) in 71% yield. Selective
replacement of the 6-O-benzyl group with O-acetyl
in 6 led to allyl 6-O-acetyl-2,3,4-tri-O-benzyl-a-
glucopyranosyl-(1 fi 3)-2-acetamido-4,6-O-benzylidene-
2-deoxy-a- -glucopyranoside (7) in only 11% yield
(Scheme 1).
The low yield of the reaction was the result of the
cleavage of the benzylidene ring with zinc chloride. So
we decided to make a similarerplacement in 4 before
formation of the disaccharide and 1,6-di-O-acetyl-2,3,4-
benzylidene-2-deoxy-a-D-glucopyranoside] (15) was
D
obtained. Signals of the O-methyl group at d 3.68 in the
1H NMR spectrum and of the carboxyl carbon atom at
d 170.5 in the 13C NMR spectrum confirm the presence
of a carboxyl group in the studied compound.
The results of the reaction shown in Scheme 2 clearly
indicate that in the reaction studied the reactivity of
glycosyl donorhas an effect on yield. In all these reac-
tions, the 1,2-cis glycoside is formed. These results seem
to be interesting because the synthesis of 1,2-cis glyco-
sides (especially oligosaccharides) is more difficult than
1,2-trans ones. A similar1,2- cis glycoside, allyl 6-O-
D
D
-
D
tri-O-benzyl-a-
D
-glucopyranose (8) was obtained in
acetyl-2,3,4-tri-O-benzyl-a-
acetamido-4,6-O-benzylidene-2-deoxy-a-
side (17), was obtained when the glycosyl donorhad the
-galacto configuration as shown in Scheme 3. The
glycosyl donor, 6-O-acetyl-2,3,4-tri-O-benzyl-a- -ga-
lactopyranosyl bromide (16), was obtained from the
appropriate 1,6-di-O-acetyl derivative. It is worthwhile
to mention, that in the preparation of this compound,
betterresults than those of the method of Kong and co-
workers gave direct transformation (without hydrolysis)
D
-galactopyranosyl-(1fi 3)-2-
good yield. Compound 8 was used as a substrate for
synthesis of three new glycosyl donors 10–12. Selective
1-O-deacetylation of 8 and reaction with trichloro-
acetonitrile gave the trichloroacetimidate 10. Reaction
of 8 with titanium tetrabromide gave 6-O-acetyl-2,3,4-
D
-glucopyrano-
D
D
tri-O-benzyl–
O-acetyl-2,3,4-tri-O-benzyl-1-thio-a-
D
-glucopyranosyl bromide (11). Phenyl 6-
-glucopyranoside
D
(12) was obtained in the reaction of 8 with thiophenol in
the presence of BF3 ethyl etherate. Condensation of 2
with 10 or 11 or 12 led to 7 in yield of 45%, 35%, and
14%, respectively. The results of the reaction of 2 with
trichloroimidate 10 indicate that the replacement of the
O-benzyl group with O-acetyl causes loweryield but
does not change the configuration of formed glycosylic
of methyl per-O-benzyl-D-galactoside in a mixture
of acetic anhydride and acetic acid in the presence of
concentrated sulfuric acid.27 Further O-deacetylation of
17 gave the compound with a free hydroxyl group at C-6
(18), which was oxidized to the uronic acid derivative 19.
Purification of 19 was not effective, so the product was
transformed into the methyl ester 20 and purified by
column chromatography.
bond. Allyl 2,3,4-tri-O-benzyl-a-
(1 fi3)-2-acetamido-4,6-O-benzylidene-2-deoxy-a-
D
-glucopyranosyl-
-gluco-
D
pyranoside (13) was obtained as the result of O-de-
acetylation of 7. Oxidation of 13 underJones condi-
There are many elements that influence the configu-
ration of a formed glycosyl bond. The most important
is neighboring group participation. A similar disaccha-
ride to the title compound, but with the b configuration,
tions23–26 gave allyl 2,3,4-tri-O-benzyl-a-
D
-glucopyrano-
syluronate-(1 fi 3)-2-acetamido-4,6-O-benzylidene-2-de-
oxy-a- -glucopyranoside (14). Its structure was con-
D
firmed by NMR spectroscopy. Additionally a small
portion of 14 was transformed into the methyl ester
with iodomethane, and allyl [methyl 2,3,4-tri-O-benzyl-
b-
D
-Glcp-(1 fi 3)-
D
-GlcpNPhth, was obtained28 when 6-
-glucopyranosyl tri-
chl-oroacetimidate was used as the glycosyl donor and
O-levulinoyl-2,3,4-tri-O-p-toluoyl-
D
a-
D
-glucopyranosyluronate-(1 fi 3)-2-acetamido-4,6-O-
4-methoxyphenyl 2-deoxy-4,6-O-benzylidene-2-phtha-
CH2OBn
O
Ph
O
O
BnO
BnO
O
+
HO
AcHN
BnO
OAll
O
CCl3
C
2
5
NH
Ph
CH2OAc
Ph
CH2OBn
O
BnO
BnO
O
BnO
O
BnO
O
O
BnO
O
BnO
O
O
O
O
AcHN
AllO
AcHN
AllO
7
6
Scheme 1.

J. Madaj et al. / Carbohydrate Research 339 (2004) 1293–1300
1295
CH2OAc
O
BnO
BnO
OAc
BnO
8
-OTf
CH2OAc
CH2OAc
CH2OAc
O
O
O
BnO
BnO
BnO
BnO
BnO
BnO
BnO
BnO
Br
BnO
O
SPh
CCl3
C
NH
12
11
AgOTf
10
TMSOTf
NIS
-OTf
CH2OAc
O
BnO
BnO
CH2OAc
CH2OAc
O
O
BnO
BnO
BnO
BnO
BnO
OTf
OTf
SPh
I
TMSOTf
BnO
BnO
2
2
CH2OAc
O
BnO
BnO
OTf
BnO
2
45%
35%
14%
R
Ph
O
BnO
BnO
O
O
BnO
O
O
AcHN
AllO
7 R = CH2OAc
13 R = CH2OH
14 R = COOH
15 R = COOCH3
Scheme 2.
BnO
on the reaction because compound 2 is not soluble in
most common organic solvents used in the synthesis of
glycosides. Compound 2 is very soluble in DMF, and we
carried out our reactions in a mixture of DMF and di-
chloromethane when used in the reactions as a solvent
only, DMF did not change the configuration of the
formed products. In our opinion the presence of the O-
benzylidene group is responsible for poor solubility of 2.
This observation seems to confirm the solubility of allyl
BnO
CH2OAc
R
Ph
O
O
BnO
O
O
2
+
BnO
BnO
O
BnO
Br
16
O
AcHN
AllO
17 R = OAc, (27%)
18 R = CH2OH
19 R = COOH
20 R = COOCH3
2-acetamido-3,5-di-O-benzyl-2-deoxy-a-D-glucopyrano-
side [A compound obtained to prepare a (1 fi 4)-disac-
charide; results will be prepared for publication] in
dichloromethane.
Scheme 3.
limido-a-
D
-glucopyranoside was the glycosyl acceptor.
Besides poorsolubility, compound 2 is rather unre-
active as a glycosyl acceptor (formation of desired di-
saccharides requires relatively long time). In our opinion
this low reactivity of 2 is the main reason of the stereo-
selectivity of the reactions. In Scheme 2 we proposed
possible explanation for a glycoside formation. In all
Because the 2-O-toluoyl group of the donor can par-
ticipate in the reaction, the 1,2-trans glycoside was ob-
tained. Anotherelement that can influence the
configuration of a formed glycosyl bond is the solvent
effect. It was difficult to state the influence of the solvent

1296
J. Madaj et al. / Carbohydrate Research 339 (2004) 1293–1300
these cases substrate a donors in reactions with pro-
moters are transformed into more reactive b byproducts.
The role of an O-triflate anion in formation of glycosylic
bond is known in the literature.29 Nucleophilic substi-
tution of the anomeric OTf group with 2 underan S N2
mechanism gave possibility to formation of the 1,2-cis
glycosides. The influence of the 6-O-acetyl group on the
configuration of a formed glycoside is proposed in the
literature.30 In our opinion, in the reactions described in
this paper, this effect is not the most important. This
conclusion could confirm our results of condensation of
ylsilyl triflate (125 lL) was added. After10 min 2,3,4,
6-tetra-O-benzyl-a- -glucopyranosyl trichloroacetimi-
D
date31 5 (0.5 g, 0.73 mmol) in dry CH2Cl2 (17 mL) was
added dropwise, and the mixture was stirred for 1 h at
)60 ꢁC and then 16 h at rt. Next Pr2EtN (0.1 mL) was
added. The solution was concentrated and co-concen-
trated with toluene to dryness. The mixture was dis-
solved in CH2Cl2, washed with aq NaHCO3 and water,
dried with MgSO4, and concentrated. The residue was
eluted from a column of silica gel with 1:3 EtOAc–tol-
20
D
uene to give 6 (0.36 g, 71%) as an oil: ½aŠ +50 (c 1,
1
2 with 2,3,4,6-tetra-O-benzyl-a-
chloroacetimidate (5) or2,3,4-tri- O-benzyl-6-O-tert-but-
yldimethylsilyl-a- -glucopyranosyl bromide (results in
D
-glucopyranosyl tri-
CHCl3); H NMR (CDCl3): d 7.4–7.24 (m, 25H, Ph),
6.25 (d, 1H, NH), 5.77 (m, 1H, @CH), 5.44 (d, 1H, J1;2
4.0, H-1II ), 5.42 (s, 1H, CHPh), 5.21 (m, 2H, @CH2),
4.94 (d, 1H, OCH2Ph), 4.87 (d, 1H, J1;2 3.2, H-1I), 4.81
D
preparation for publication). The presence of a 6-O-
benzyl or6- O-tert-butyldimethylsilyl group in the mol-
ecule of the glycosyl donordoes not change the product
configuration.
(d, 1H, OCH2Ph), 4.70 (d, 1H, OCH2Ph), 4.5 (m, 3H,
0
OCH2Ph), 4.39 (m, 1H, H-2I), 4.24 (m, 3H, H-3I, H-6II ,
0
OCH2Ph), 4.20 (m, 2H, H-6I , OCH2), 3.85 (m, 4H, H-
In summary, allyl 2-acetamido-4,6-O-benzylidene-2-
-glucopyranoside (2), even if it is difficultly
3II, H-4I, H-5II, OCH2), 3.77 (m, 2H, H-6I, H-6II), 3.5
deoxy-a-
D
(m, 2H, H-2II, H-5I), 3.19 (dd, 1H, J3;4 10.0, J4;5 10.4, H-
13
soluble and unreactive, is interesting as the glycosyl
acceptor because it affords possibilities for preparation
of 1,2-cis glycosides found in nature.
4II), 1.86 (s, 3H, NCOCH3).
C NMR d 170.69
(NCOCH3), 138.89 (@CH), 138.4–126.64 (Ph), 118.07
(@CH2), 102.44 (CHPh), 97.24 (C-1I), 96.36 (C-1II),
83.10 (C-3II), 81.88 (C-4I), 78.73 (C-2II), 77.99 (C-4II),
75.97 (OCH2Ph), 74.75 (OCH2Ph), 74.17 (OCH2Ph),
72.45 (C-3I), 71.03 (OCH2Ph), 70.49 (C-6II), 70.31 (C-
5I), 69.26 (C-6I), 68.7 (OCH2), 62.92 (C-5II), 52.27 (C-
2I), 23.26 (NHCOCH3). Anal. Calcd forC 52H57NO11: C,
71.64; H, 6.54; N, 1.6. Found: C, 70.95; H, 6.87; N, 1.01.
3. Experimental
3.1. General methods
All reactions were carried out in commercially available
dry solvents (Fluka, water <0.005%). Thin-layer chro-
matography was performed with E. Merck pre-coated
Silica Gel 60 F-254 plates, and detection of compounds
was achieved by charring after spraying with 5% H2SO4
in EtOH. Column chromatography was carried out with
3.3. Phenyl 6-O-acetyl-2,3,4-tri-O-benzyl-1-thio-a-D-
glucopyranoside (11)
Boron trifluoride-diethyl ether (0.3 mL) was added to a
solution of 1,6-di-O-acetyl-2,3,4-tri-O-benzyl-a- -glu-
D
copyranose22 8 (0.87 g, 1.6 mmol) and PhSH (0.3 mL,
2.94 mmol) in dry CH2Cl2 (15 mL), and the mixture was
stirred for 20 h at rt. Next the mixture was diluted with
CH2Cl2, washed with aq NaHCO3 and water, dried with
MgSO4, and concentrated. The residue was crystallized
from MeOH to give compound 11 (0.31 g, 32%), mp
1
Kieselgel 60 Silica Gel (E. Merck, <200 mesh). H and
13C NMR spectra were recorded at 25 ꢁC with a Varian
Mercury spectrometer at 400 and 100 MHz, respectively,
with Me4Si as internal standard. Assignments were
based on homonucleardecoupling expeimr ents and
homo- and heteronuclear correlation. Mass spectra were
measured using MALDI-TOF with a-cyano-4-hy-
droxycinnamic acid (CCA) as the matrix. Optical rota-
tions were measured with a JASCO J-20 polarimeter.
Elemental analyses were carried out with a Carlo Erba
apparatus.
1
112.6–113.0 ꢁC; anomer b was not isolated. H NMR
(CDCl3): d 7.47–7.26 (m, 20H, Ph), 5.60 (d, 1H, J1;2 5.0,
H-1), 5.02 (d, 1H, OCH2Ph), 4.88 (d, 1H, OCH2Ph),
4.82 (d, 1H, OCH2Ph), 4.78 (d, 1H, OCH2Ph), 4.69 (d,
1H, OCH2Ph), 4.57 (d, 1H, OCH2Ph), 4.03 (m, 1H, H-
0
5), 4.28 (dd, 1H, J5:6 5.2, J6;6 11.6, H-6), 4.07 (dd, 1H,
J5:6 2.0, H-60), 3.93 (t, 1H, J3;4 10.0, H-3), 3.87 (dd, 1H,
0
3.2. Allyl 2,3,4,6-tetra-O-benzyl-a-
(1 fi 3)-2-acetamido-4,6-O-benzylidene-2-deoxy-a-
glucopyranoside (6)
D
-glucopyranosyl-
D-
J2;3 9.6, H-2), 3.51 (dd, 1H, J4;5 8.8, H-4) 1.96 (s, 3H,
COOCH3). 13C NMR d 170.86 (COOCH3), 138.68–
127.54 (Ph), 86.93 (C-1), 82.63 (C-3), 79.93 (C-2), 77.34
(C-4), 76.05 (OCH2Ph), 75.31 (OCH2Ph), 72.74
Allyl
2-acetamido-4,6-O-benzylidene-2-deoxy-a-D-
glucopyranoside 2 (0.2 g, 0.58 mmol, prepared per the
Warren and Jeanloz procedure11) in dry DMF (17 mL)
was stirred at )60 ꢁC under dry nitrogen, and trimeth-
Indices according to Nomenclature of Carbohydrates (Recommen-
dations 1996), Carbohydr. Res. 1997, 297, 1–92.

J. Madaj et al. / Carbohydrate Research 339 (2004) 1293–1300
1297
(OCH2Ph), 69.78 (C-5), 63.31 (C-6), 20.10 (COOCH3).
Anal. Calcd forC 35H36O6S: C, 71.92; H, 6.16; S, 5.48.
Found: C, 71.93; H, 6.36; S, 5.24.
of silica gel with 1:1 EtOAc–toluene to give 7 (49 mg,
1
20
D
14%) as an oil: ½aŠ +65.9 (c 1.4, CHCl3); H NMR
(CDCl3): d 7.4–7.1 (m, 20H, Ph), 6.0 (d, 1H, NH), 5.89
(m, 1H, @CH), 5.49 (d, 1H, J1;2 3.6 Hz, H-1II), 5.44 (s,
1H, CHPh), 5.32 (m, 2H, @CH2), 4.97 (d, 1H,
OCH2Ph), 4.89 (d, 1H, J1;2 4.0 Hz, H-1I), 4.87 (d, 1H,
OCH2Ph), 4.77 (d, 1H, OCH2Ph), 4.53 (m, 2H,
OCH2Ph), 4.48 (m, 1H, H-2I), 4.3 (d, 1H, OCH2Ph),
4.27–4.16 (m, 4H, H-3I, H-5II, H-6II, OCH2), 4.12 (m,
2H, H-4I, H-6II;), 4.04 (m, 1H, OCH2), 3.95 (m, 3H, H-
3II, H-6I, H-6I;), 3.77 (m, 1H, J4;5 9.6 Hz, J5;6 10.4 Hz, H-
5I), 3.38 (dd, 1H, J2;3 9.2, H-2II), 3.25 (t, 1H, J3;4 9.2 Hz,
J4;5 9.6 Hz, H-4II), 2.08 (s, 3H,COCH3), 2.02 (s, 3H,
NCOCH3). 13C NMR d 171.18 (COCH3), 170.35
(NCOCH3), 138.86 (@CH), 138.27–126.61 (Ph), 118.72
(@CH2), 102.41 (CHPh), 97.66 (C-1I), 96.09 (C-1II),
83.22 (C-3II), 81.48 (C-4I), 79.32 (C-2II), 78.64 (C-4II),
75.9 (OCH2Ph), 74.77 (OCH2Ph), 72.06 (C-5II), 71.15
(OCH2Ph), 69.2 (C-5I), 68.91 (OCH2), 68.31 (C-3I),
64.58 (C-6II), 62.99 (C-6I), 5.86 (C-2I), 23.31 (COCH3),
21.15 (NHCOCH3); MALDITOF-MS: calcd for
C47H53NO12: 823.92 [M]. Found: 846.3 [M+Na]þ.
3.4. 6-O-acetyl-2,3,4-tri-O-benzyl-a-D-glucopyranosyl
bromide (12)
To a solution of 8 (0.6 g, 1.1 mmol) in a mixture of EtOAc
(1.1 mL), and CH2Cl2 (11 mL), TiBr4 (1.2 g, 3.3 mmol)
was added, and the mixture was stirred for 2 h at rt. The
mixture was diluted with CH2Cl2, washed with iced-cold
water, dried with MgSO4, and concentrated. The syrupy
residue (0.48 g) was immediately used for next reaction.
3.5. Allyl 6-O-acetyl-2,3,4-tri-O-benzyl-a-
anosyl-(1 fi 3)-2-acetamido-4,6-O-benzylidene-2-deoxy-
a- -glucopyranoside (7)
D-glucopyr-
D
3.5.1. Procedure (a). A mixture of allyl 2-acetamido-4,6-
O-benzylidene-2-deoxy- -glucopyranoside (2) (0.2 g,
0.58 mmol), 2,4,6-collidine (0.2 mL, 1.6 mmol), and
D
ꢁ
molecularsieves 4 A (1 g) in dry DMF (15 mL) was
stirred at rt under dry nitrogen, and silver triflate (0.4 g,
1.6 mmol) was added. After10 min, 6- O-acetyl-2,3,4-tri-
-glucopyranosyl bromide 12 (0.48 g) in dry
3.6. Allyl 2,3,4-tri-O-benzyl-a- -glucopyranosyl-(1 fi 3)-
2-acetamido-4,6-O-benzylidene-2-deoxy-a-D-glucopyr-
anoside (13)
D
O-benzyl-a-
D
CH2Cl2 (17 mL) was added dropwise, and the mixture
was stirred for 20 h at rt. Next Pr2EtN (0.1 mL) was
added. The mixture was concentrated and co-concen-
trated with toluene to dryness. The residue was dis-
solved in CH2Cl2, filtrated through Celite, washed with
aq NaHCO3 and water, dried with MgSO4, and con-
centrated. The residue was eluted from a column of
silica gel with 1:1.5 EtOAc–toluene to give 7 (0.165 g,
35%) as an oil.
The solution of 7 (0.364 g, 0.44 mmol) in saturated
methanolic ammonia was stirred for 1 day at rt. Next the
solution was concentrated to dryness to give 13 (0.33 g,
20
1
95%) as an oil: ½aŠ +75.5 (c 1.0, CHCl3); H NMR
D
(CDCl3): d 7.4–7.1 (m, 20H, Ph), 6.23 (d, 1H, NH), 5.9
(m, 1H, @CH), 5.43 (m, 2H, H-1II, CHPh), 5.37 (m, 2H,
@CH2), 4.96 (d, 1H, OCH2Ph), 4.89 (d, 1H, J1;2 3.6 Hz,
H-1I), 4.68 (d, 2H, OCH2Ph), 4.51 (m, 2H, OCH2Ph),
4.45 (dd, 1H, J2;3 9.6 Hz, H-2I), 4.29 (d, 1H, OCH2Ph),
4.25–4.15 (m, 3H, H-3I, H-6II;, OCH2), 4.02–3.73 (m,
7H, H-3II, H-4I, H-5II, H-6I, H-6I;, H-6II, OCH2), 3.39
(dd, 1H, J1;2 4.0 Hz, J2;3 9.6 Hz, H-2II), 3.22 (dd, 1H, J3;4
8.8 Hz, H-4II), 2.02 (s, 3H, NCOCH3). 13C NMR d
170.65 (NCOCH3), 138.95 (@CH), 138.45–126.6 (Ph),
118.31 (@CH2), 102.41 (CHPh), 97.55 (C-1I), 96.39 (C-
1II), 83.11 (C-3II), 81.68 (C-4I), 79.03 (C-2II), 78.21 (C-
4II), 75.87 (OCH2Ph), 74.77 (OCH2Ph), 72.78 (C-3I),
71.28 (C-5II), 71.02 (OCH2Ph), 69.23 (C-6II), 68.87
(OCH2), 62.98 (C-5I), 62.94 (C-6I), 52.11 (C-2I), 23.26
(NHCOCH3). Anal. Calcd orC 45H51NO11: C, 69.14; H,
6.53; N, 1.79. Found: C, 69.0; H, 6.64; N, 2.07.
3.5.2. Procedure (b). A mixture of 2 (0.17 g, 0.49 mmol),
6-O-acetyl-2,3,4,-tri-O-benzyl-a-D-glucopyranosyl tri-
chloroacetimidate32 10 (0.39 g, 0.61 mmol), and trim-
ethylsilyl triflate (160 lL) was stirred in dry DMF
(20 mL), and CH2Cl2 (20 mL) at )60 ꢁC for1 h, and then
at rt for 16 h as described above for preparation from 6.
Subsequent chromatography (1:2 EtOAc–toluene)
afforded 7 (0.18 g, 45%) as an oil.
3.5.3. Procedure (c). The mixture of
0.42 mmol), phenyl 6-O-acetyl-2,3,4-tri-O-benzyl-1-thio-
a- -glucopyranoside 11 (0.3 g, 0.51 mmol), NIS (0.12 g,
2 (0.15 g,
D
ꢁ
0.51 mmol), and MS 4 A (1 g) in a mixture of DMF
(7 mL)–CH2Cl2 (5 mL)–EtOAc (5 mL) was stirred at
0 ꢁC under dry nitrogen. After 5 min trimethylsilyl tri-
flate (30 lL) was added, and the mixture was stirred for
0.5 h at 0 ꢁC and then for 16 h at rt. The mixture was
concentrated and co-concentrated with toluene to dry-
ness. The residue was dissolved in CH2Cl2, filtered,
washed with aq NaHCO3 and water, dried with MgSO4,
and concentrated. The residue was eluted from a column
3.7. Allyl 2,3,4-tri-O-benzyl-a-D-glucopyranosyluronate-
(1 fi 3)-2-acetamido-4,6-O-benzylidene-2-deoxy-a-
glucopyranoside (14)
D-
The mixture containing CrO3 (0.47 g, 4.7 mmol), concd
H2SO4 (0.39 mL) and water(1.9 mL) was added dorp-
wise to a solution of 13 (0.41 g, 0.53 mmol) in acetone

1298
J. Madaj et al. / Carbohydrate Research 339 (2004) 1293–1300
(15 mL). Then the reaction mixture was poured into ice-
waterand extarcted with EtOAc. The EtOAc solution
was washed with water, dried with MgSO4, and con-
centrated. The residue was eluted from a column of
3.9. 1,6-Di-O-acetyl-2,3,4-tri-O-benzyl-D-galactopyra-
nose33 (16)
Concd H2SO4 (1.53 mL) was added dropwise to a stirred
solution of methyl 2,3,4,6-tetra-O-benzyl-a- -galacto-
silica gel with 10:9:1 CH2Cl2–EtOAc–AcOH to give 14
D
20
D
(0.3 g, 72%) as an oil: ½aŠ +64.5 (c 1, CHCl3); 1H NMR
pyranoside (8.2 g, 14.8 mmol) in a mixture of AcOH
(39 mL) and Ac2O (39 mL) at 0 ꢁC. After1 h water
(100 mL) was added, and the mixture was extracted with
CH2Cl2. The organic layer was washed with aq
NaHCO3 water, dried with MgSO4, and concentrated to
(CDCl3): d 7.38–7.08 (m, 20H, Ph), 6.37 (d, 1H, NH),
5.89 (m, 1H, @CH), 5.56 (d, 1H, J1;2 3.2 Hz, H-1II), 5.43
(s, 1H, CHPh), 5.29 (m, 2H, @CH2), 4.92 (d, 1H,
OCH2Ph), 4.84 (d, 1H, J1;2 4.0 Hz, H-1I), 4.74 (m, 2H,
OCH2Ph), 4.55 (m, 3H, H-2I, OCH2Ph), 4.35 (d, 1H, J4;5
10 Hz, H-5II), 4.49 (d, 1H, OCH2Ph), 4.29 (d, 1H,
OCH2Ph), 4.24 (m, 1H, H-3I), 4.17 (m, 1H, OCH2), 3.99
(m, 1H, OCH2), 3.90 (m, 4H, H-3II, H-4I, H-6I, H-6I;),
3.74 (m, 1H, J4:5 9.6 Hz, J5;6 10.4 Hz, H-5I), 3.64 (dd,
1H, J3;4 9.2, J4;5 9.6, H-4II), 3.44 (dd, 1H, J2;3 10, H-2II),
2.09 (s, 3H, NCOCH3). 13C NMR d 173.57 (COOH),
171.49 (NCOCH3), 138.77 (@CH), 138.17–126.54 (Ph),
118.66 (@CH2), 102.22 (CHPh), 97.45 (C-1I), 96.88 (C-
1II), 82.78 (C-3II), 80.78 (C-4I), 78.99 (C-4II), 78.37 (C-
2II), 75.97 (OCH2Ph), 74.94 (OCH2Ph), 73.37 (C-3I),
71.55 (OCH2Ph), 70.54 (C-5II), 69.12 (C-5I), 68.75
(OCH2), 62.74 (C-6I), 51.86 (C-2I), 23.39 (NCOCH3);
MALDITOF-MS: Calcd forC 45H49O12N: 795.87 [M].
Found: 818.2 [M+Na]þ.
1
give 16 (6.5, 82%) as an oil: H NMR (CDCl3): d 7.4–
7.25 (m, 15H, Ph), 6.39 (d, 1H, J1;2 3.6 Hz, H-1), 4.98 (d,
1H, OCH2Ph), 4.88 (d, 1H, OCH2Ph), 4.76 (d, 1H,
OCH2Ph), 4.71 (m, 2H, OCH2Ph), 4.62 (d, 1H,
OCH2Ph), 4.18 (dd, 1H, J1;2 10.0 Hz, J2;3 10.0 Hz, H-2),
0
4.13 (dd, 1H, J5:6 6.4 Hz, J6;6 10.8 Hz, H-6), 4.07 (dd,
1H, J5:6 6.0 Hz, H-60), 4.02 (m, 1H, H-5) 3.92 (bd, 1H,
0
H-4), 3.88 (dd, 1H, J3;4 3.2 Hz, H-3), 2.12 (s, 3H, CO-
OCH3), 1.98 (s, 3H, COOCH3). 13C NMR d 170.78
(COOCH3), 169.63 (COOCH3), 138.68–127.63 (Ph),
90.84 (C-1), 78.78 (C-3), 75.55 (C-2), 74.89 (OCH2Ph),
74.37 (C-4), 73.62 (OCH2Ph), 73.61 (OCH2Ph), 71.01
(C-5), 63.27 (C-6), 21.35 (COOCH3), 21.02 (COOCH3).
3.10. 6-O-Acetyl-2,3,4-tri-O-benzyl-a-D-galactopyrano-
syl bromide33 (17)
3.8. Allyl (methyl 2,3,4-tri-O-benzyl-a-
luronate-(1 fi 3)-2-acetamido-4,6-O-benzylidene-2-
deoxy-a- -glucopyranoside) (15)
D-glucopyranosy-
Compound 17 was prepared from 16 (1.09 g, 2.03 mmol)
as described for the preparation of 12, yielding (0.93 g).
The syrupy product was immediately used for next
reaction.
D
To a solution of 14 (50 mg, 0.07 mmol) in dry DMF
(1.5 mL) was added KHCO3 (50 mg, 0.5 mmol) and
CH3I (40 lL, 0.64 mmol). The mixture was stirred 1 h at
0 ꢁC. After evaporation, the residue was dissolved in
3.11. Allyl 6-O-acetyl-2,3,4-tri-O-benzyl-a-
anosyl-(1 fi 3)-2-acetamido-4,6-O-benzylidene-2-deoxy-
a- -glucopyranoside (18)
D-galactopyr-
CH2Cl2, washed with water, dried with MgSO4, and
D
20
concentrated gave 15 (35 mg, 62%) as an oil: ½aŠ +63.2
D
(c 1, CHCl3); 1H NMR (CDCl3): d 7.4–7.1 (m, 20H, Ph),
5.93 (m,1H, @CH), 5.85 (d, 1H, NH), 5.54 (d, 1H, J1;2
3.6 Hz, H-1II), 5.39 (s, 1H, CHPh), 5.29 (m, 2H, @CH2),
4.92 (d, 1H, OCH2Ph), 4.88 (d, 1H, J1;2 3.6 Hz, H-1I),
4.76 (d, 1H, OCH2Ph), 4.62 (m, 2H, OCH2Ph), 4.55–
4.48 (m, 2H, H-2I, OCH2Ph), 4.38 (d,1H, J4;5 10.4 Hz,
H-5II), 4.27 (d, 1H, OCH2Ph), 4.18 (m, 2H, H-3I,
OCH2), 4.02 (m, 1H, OCH2), 3.88 (m, 4H, H-3II, H-4I,
H-6I, H-6I;), 3.74 (dd, 1H, J4;5 10.0 Hz, J5;6 10.4 Hz, H-
5I), 3.68 (s, 3H, COOCH3), 3.63 (t, 1H, J3;4 9.2 Hz, H-
4II), 3.43 (dd, 1H, J2;3 10 Hz, H-2II), 2.08 (s, 3H,
NCOCH3). 13C NMR d 170.5 (COOCH3), 170.08
(NCOCH3), 138.8 (@CH), 138.2–126.5 (Ph), 118.49
(@CH2), 102.29 (CHPh), 97.45 (C-1I), 96.99 (C-1II),
82.99 (C-3II), 80.9 (C-4I), 78.99 (C-4II), 78.37 (C-2II),
75.98 (OCH2Ph), 74.77 (OCH2Ph), 73.17 (C-3I), 71.42
(OCH2Ph), 70.92 (C-5II), 69.17 (C-5I), 68.83 (OCH2),
62.74 (C-6I), 52.65 (COOCH3), 51.79 (C-2I), 23.51
(NCOCH3); MALDITOF-MS: Calcd forC 46H51NO12:
809.9 [M]. Found: 832.4 [M+Na]þ.
Glycosylation of 2 (0.36 g, 1.03 mmol) with bromide 17
(0.9 g) in dry DMF (15 mL) and CH2Cl2 (15 mL) pro-
moted by silver triflate (0.75 g, 2.9 mmol) was carried
out as described above for preparation of 7 (procedure
b). Subsequent purification of the product by column
chromatography (1:1.5 EtOAc–toluene) afforded 18
20
D
(0.23 g, 27%) as an oil: ½aŠ +66.7 (c 1, CHCl3); 1H
NMR (CDCl3): d 7.4–7.1 (m, 20H, Ph), 5.98 (d, 1H,
NH), 5.9 (m, 1H, @CH), 5.43 (d, 1H, J1;2 4.0, H-1II),
5.44 (s, 1H, CHPh), 5.3 (m, 2H, @CH2), 4.97 (d, 1H,
OCH2Ph), 4.87 (d, 1H, J1;2 3.6 Hz, H-1I), 4.85 (d, 1H,
OCH2Ph), 4.74 (d, 1H, OCH2Ph), 4.53 (m, 2H,
OCH2Ph), 4.48 (m, 1H, H-2I), 4.29 (d, 1H, OCH2Ph),
4.26–4.22 (m, 2H, H-6I, H-6II), 4.19 (m, 1H, OCH2),
4.16–4.07 (m, 3H, H-3I, H-5II, H-6II;), 4.03 (m, 1H,
OCH2), 3.91 (m, 3H, H-3II, H-4II, H-5I), 3.76 (m, 1H,
0
J6;6 10.0 Hz, H-6II ), 3.38 (dd, 1H, J2;3 10.0 Hz, H-2II),
0
3.25 (t, 1H, J3;4 9.2, J4;5 9.6, H-4II), 2.08 (s, 3H,COCH3),
2.02 (s, 3H, NCOCH3). 13C NMR d 171.19 (COCH3),
170.33 (NCOCH3), 138.89 (@CH), 138.29–125.51 (Ph),

J. Madaj et al. / Carbohydrate Research 339 (2004) 1293–1300
1299
118.74 (@CH2), 102.44 (CHPh), 97.71 (C-1I), 96.08 (C-
1II), 83.28 (C-5I), 81.49 (C-3II), 78.68 (C-2II), 77.99 (C-
4II), 75.93 (OCH2Ph), 74.8 (OCH2Ph), 72.08 (C-3I),
71.19 (OCH2Ph), 69.23 (C-6I), 68.93 (OCH2), 68.33 (C-
5II), 64.61 (C-6II), 62.99 (C-4II), 5.88 (C-2I), 23.35
(COCH3), 21.16 (NHCOCH3); MALDITOF-MS: Calcd
forC 47H53 NO12: 823.92 [M]. Found: 846.3 [M+Na]þ.
77.98 (C-2II), 78.99 (C-4II), 76.54 (C-3I), 74.97 (C-6I),
74.91 (C-3II), 74.82 (OCH2Ph), 73.88 (OCH2Ph), 71.87
(OCH2Ph), 71.27 (C-5II), 69.06 (C-5I), 68.76 (OCH2),
62.51 (C-4II), 52.36 (COOCH3), 51.79 (C-2I), 23.38
(NCOCH3); MALDITOF-MS: Calcd forC 46H51NO12:
809.9 [M]. Found: 832.2 [M+Na]þ.
3.12. Allyl 2,3,4-tri-O-benzyl-a-D-galactopyranosyl-
Acknowledgements
(1 fi 3)-2-acetamido-4,6-O-benzylidene-2-deoxy-a-
glucopyranoside (19)
D-
This work was supported by a research grants BW/8000-
5-0237-3 and DS/8361-4-0134-3 from the Polish State
Committee forScientific Research (KBN).
Compound 19 was prepared from 18 (0.2 g, 0.19 mmol)
as described for the preparation of 13, yielding (0.16 g,
20
D
1
85%) as a syrup: ½aŠ +74.2 (c 1, CHCl3); H NMR
(CDCl3): d 7.38–7.1 (m, 20H, Ph), 6.26 (d, 1H, NH),
5.87 (m, 1H, @CH), 5.55 (d, 1H, J1;2 3.6 Hz, H-1II), 5.38
(s, 1H, CHPh), 5.22 (m, 2H, @CH2), 4.87 (m, 2H,
OCH2Ph, H-1I), 4.69 (d, 1H, OCH2Ph), 4.52 (m, 2H,
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ꢀ
ꢀ
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0
4.31 (d, 1H, OCH2Ph), 4.22–4.11 (m, 4H, H-3I, H-6I , H-
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Calcd
for
C45H51NO11: 781.89 [M]. Found: 804.2 [M+Na]þ.
3.13. Allyl (methyl 2,3,4-tri-O-benzyl-a-
anosyluronate-(1 fi 3)-2-acetamido-4,6-O-benzylidene-2-
deoxy-a- -glucopyranoside) (20)
D-galactopyr-
D
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D
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5.35 (s, 1H, CHPh), 5.23 (m, 2H, @CH2), 4.85 (m, 3H,
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