An access to various sulfation patterns in dermatan sulfate: Chemical syntheses of sulfoforms of trisaccharide methyl glycosides

Article abstract of DOI:10.1016/S0008-6215(02)00060-5

The syntheses are reported for the first time of α-L-IdopA2SO₃-(1→3)-β-D-GalpNAc4SO₃- (1→4)-α-L-IdopA2SO₃-(1→OMe), its disulfated analogue α-L-IdopA2SO₃-(1→3)-β-D-GalpNAc-(1→4)- α-L-IdopA2SO₃-(1→OMe), and of β-D-GalpNAc4SO₃-(1→4)-α-L-IdopA2SO₃- (1→3)-β-D-GalpNAc4SO₃-(1→OMe), which represent structural fragments of dermatan sulfate, unavailable directly by chemical or enzymatic degradation of the glycosaminoglycan polymer. These molecules were readily obtained from a pair of key disaccharide intermediates, in which the relative difference of stability of the D-GalNAc 4-hydroxy protecting groups (acetate or pivalate) toward saponification conditions allowed access to various sulfoforms from a common precursor. For the preparation of these blocks, the 4-O-pivaloyl-D-galacto moiety was readily obtained through a one-pot stereospecific intramolecular nucleophilic displacement on an easily available 3-O-pivaloyl-D-gluco precursor, and the L-IdoA moiety through selective radical oxidation at C-6 of a L-ido 4,6-diol derivative with oxoammonium salts.

Full text of DOI:10.1016/S0008-6215(02)00060-5

Carbohydrate Research 337 (2002) 673–689
www.elsevier.com/locate/carres
An access to various sulfation patterns in dermatan sulfate:
chemical syntheses of sulfoforms of trisaccharide methyl glycosides
Nadine Barroca, Jean-Claude Jacquinet*
Institut de Chimie Organique et Analytique, UFR Faculte´ des Sciences, UMR CNRS 6005, Uni6ersite´ d’Orle´ans, BP 6759,
F-45067 Orle´ans, France
Received 10 January 2002; accepted 15 February 2002
Abstract
The syntheses are reported for the first time of a-
its disulfated analogue a- -IdopA2SO3-(1“3)-b- -GalpNAc-(1“4)-a-
4)-a- -IdopA2SO3-(1“3)-b- -GalpNAc4SO3-(1“OMe), which represent structural fragments of dermatan sulfate, unavailable
directly by chemical or enzymatic degradation of the glycosaminoglycan polymer. These molecules were readily obtained from a
pair of key disaccharide intermediates, in which the relative difference of stability of the -GalNAc 4-hydroxy protecting groups
(acetate or pivalate) toward saponification conditions allowed access to various sulfoforms from a common precursor. For the
preparation of these blocks, the 4-O-pivaloyl- -galacto moiety was readily obtained through a one-pot stereospecific intramolec-
ular nucleophilic displacement on an easily available 3-O-pivaloyl- -gluco precursor, and the -IdoA moiety through selective
radical oxidation at C-6 of a -ido 4,6-diol derivative with oxoammonium salts. © 2002 Elsevier Science Ltd. All rights reserved.
L
-IdopA2SO3-(1“3)-b-
D
-GalpNAc4SO3-(1“4)-a-
L
-IdopA2SO3-(1“OMe),
L
D
L
-IdopA2SO3-(1“OMe), and of b-
D
-GalpNAc4SO3-(1“
L
D
D
D
D
L
L
Keywords: Dermatan sulfate trisaccharides; Sulfoforms; Glycosylation reactions; 2-Deoxy-2-trichloroacetamido-D-galacto derivatives
1. Introduction
responsiveness,4 its possible anti-oncogenic role,5 or its
interaction of high specificity to hepatocyte growth
factor.6
Dermatan sulfate (DS) belongs to the proteoglycan
family, and is found in a wide variety of animal tissues.
It is a microheterogeneous, polydisperse, linear copoly-
mer composed of disaccharide repeating units of 2-acet-
Homogeneous, structurally characterized DS frag-
ments might be useful for a better understanding of
these biological properties and of the specificity of
enzymes acting on DS. Such compounds are not easily
accessible by enzymatic or chemical degradation of the
polymer, and have generally to be chemically synthe-
sized. To date, several syntheses of DS fragments hav-
ing an even number of saccharide residues have been
reported, such as those of disaccharide derivatives hav-
amido-2-deoxy-
-iduronic acid (
GlcpA), with sulfate groups most commonly present at
C-4 of -GalNAc and occasionally at C-2 of -IdoA
D
-galactose
(
D
-GalpNAc) and of
L
L
-IdopA) or
D
-glucuronic acid (
D-
D
L
residues, but other sulfation patterns depending on its
origin are known. The most studied biological activities
of DS are its anticoagulant1 and antithrombotic2 prop-
erties, where it displays less hemorragic effects than
heparin.3 But many other implications of DS in impor-
tant biological processes, even if still poorly understood
at the molecular level, are known, such as its potent
mediator effect of fibroblast growth factor-2 (FGF-2)
ing
D L
-GalNAc7–9 or -IdoA10 residues at the reducing
end, or of an hexasaccharide,11,12 the postulated mini-
mal sequence1 binding to heparin co-factor II (HCII).
Recently, tri- and pentasaccharide fragments exclusively
sulfated at C-4 of the D-GalNAc residues were obtained
by controlled depolymerization of porcine intestinal
mucosal DS using chondroitinase ABC, and removal of
the D4,5-unsaturated nonreducing L-IdoA residue with
* Corresponding author. Tel.: +33-238-417072; fax: +33-
238-417281.
E-mail address: jean-claude.jacquinet@univ-orleans.fr (J.-
C. Jacquinet).
mercuric acetate.13
The current study describes, for the first time, a
versatile approach to various sulfoforms of DS trisac-
0008-6215/02/$ - see front matter © 2002 Elsevier Science Ltd. All rights reserved.
PII: S0008-6215(02)00060-5

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N. Barroca, J.-C. Jacquinet / Carbohydrate Research 337 (2002) 673–689
charide derivatives having either
L
-IdoA (1, 2) or
D
-
other ester groups (i.e., acetate or benzoate). It is also
an excellent candidate for intramolecular nucleophilic
displacement reactions leading to inversion of configu-
GalNAc (3) residues at the reducing end (Scheme 1).
All compounds were prepared as their methyl gly-
cosides, with the same anomeric configuration as in the
natural polymer, in which the methyl group was intro-
duced as a marker for NMR studies. Such molecules
are not only useful for the studies of interactions with
biologically important proteins, but also for a better
understanding of DS biosynthesis, and particularly for
the characterization of the specific GalNAc 4-O-
sulfotransferase.14
ration on sugar carbons such as in
transformations.17 Thus, its use as a temporary protec-
tion at O-4 of -GalNAc residues might allow to
D
-gluco“D-galacto
D
prepare the corresponding 4-sulfate or 4-OH species
from a common precursor. But care has to be taken in
order to ensure complete removal of this group at the
final deprotection step.16 Consequently, for the synthe-
ses of the target trisaccharide derivatives 1–3, a couple
of key disaccharide intermediates (20, 21) were de-
signed, which may, in turn, be bond-disconnected into
a glycosyl donor (13), and a pair of
acceptors (7, 8) readily obtained from a common pre-
cursor (6) having the -gluco configuration (Scheme 1).
Preparation of acceptors 7 and 8 was achieved in a
straightforward manner as follows (Scheme 2). Pivaloy-
lation of easily available 4-methoxyphenyl 4,6-O-ben-
D-galacto glycosyl
2. Results and discussion
D
As far as DS fragments contain sulfate esters, the
usual strategy8–12 for their chemical synthesis is to
temporarily protect the hydroxy groups which would
carry these substituents with ester groups, and those
which should remain free with permanent benzyl ethers.
Generally, all ester groups, regardless of their nature,
are removed at the same time, without seeking for a
possible selective removal. Pivaloyl ester is known to be
much more resistant to basic conditions15,16 than are
zylidene-2-deoxy-2-trichloroacetamido-b-D-glucopyrano
side (4)10 gave the crystalline ester 5 in 82% yield.
Regioselective reductive cleavage of the benzylidene
acetal in 5 with triethylsilane and trifluoroacetic acid18
afforded the alcohol 6 in 91% yield. It is worth noting
Scheme 1.

N. Barroca, J.-C. Jacquinet / Carbohydrate Research 337 (2002) 673–689
675
Scheme 2.
and 15 in 78 and 79% yields, respectively. The 1H NMR
spectra for 14 and 15 (Table 3) are in agreement with
that direct reductive cleavage on alcohol 4, followed by
regioselective pivaloylation at O-3 of the intermediate
proceeded with lower stereospecificity and overall yield.
One-pot inversion of configuration with concomitant
protection at C-4 was readily achieved by treatment of
the expected structures, and show, for the L-ido moi-
1
eties, a significant staggering from the C4 conforma-
tion observed for monomers 9–13. Treatment of 14 and
15 with aqueous acetic acid at 100 °C afforded the
corresponding 4II,6II-diols, on which regioselective oxi-
dation at C-6 without protection at C-4 was attempted.
Selective oxidation of primary alcohol in sugar 4,6-diol
by using the oxyammonium ion-catalyzed procedure
under phase-transfer conditions has been reported,20,21
and was successfully applied in our group for the
6
with triflic anhydride and pyridine in 1,2-
dichloroethane at −15 °C, followed by in situ addition
of water,17 and heating at 80 °C to give, via the corre-
sponding 3,4-hemiorthoester intermediate, the 4-O-pi-
valoyl- -galacto acceptor 7 in 82% overall yield, the
D
structure of which was easily deduced from its 1H
NMR spectrum (Table 1). With the aim of avoiding
deprotection problems with the pivaloyl groups at the
end of the syntheses, the 4-O-acetyl analogue 8 was
prepared, at least as a spare derivative. Thus, Zemple`n
transesterification of 7 gave the corresponding diol,
which was directly treated with trimethyl orthoacetate
preparation of
thesis of chondroitin sulfate
ever, frustrating results were obtained starting from
-ido derivatives,20 as was the case with the diols
D
-glucuronic acid derivatives in the syn-
D
oligosaccharides.22 How-
L
derived from 14 and 15. Nevertheless, when these diols
were treated at 0 °C with a catalytic amount of 2,2,6,6-
tetramethyl-1-piperidinyloxy, free radical (TEMPO), in
the presence of stable calcium hypochlorite21 as a co-
and 10-D,L-camphorsulfonic acid in toluene to give the
3,4-orthoester intermediate, which was, in turn, regiose-
lectively opened19 in acidic medium to give the crys-
talline acceptor 8 in 86% yield.
The
(Scheme 3). Treatment of known10 1,2,4,6-tetra-O-ben-
zoyl-3-O-benzyl-b- -idopyranose (9) with 4-methoxy-
L
-ido donor 13 was then prepared as follows
Table 1
1H NMR data: carbohydrate ring protons for monosaccha-
ride derivatives 5–8 a
L
phenol in dichloromethane, and in the presence of
trimethylsilyl triflate (1 equiv), afforded glycoside 10 in
nearly quantitative yield, the anomeric configuration of
Proton
5
6
7
8
1
H-1
J1,2
H-2
J2,3
H-3
J3,4
H-4
J4,5
H-5
J5,6a
J5,6b
H-6a
J6a,6b
H-6b
NH
J2,NH
4.90
8.4
4.50
10.3
5.61
5.00
8.2
4.28
10.7
5.43
8.8
3.82
9.7
3.79
5.20
8.5
3.91
11.2
4.36
3.5
5.40
0.9
3.97
6.3
6.3
3.64
5.17
8.4
3.98
10.9
4.36
which was deduced from its H NMR spectrum (Table
2). Transesterification of 10, followed by treatment with
benzaldehyde and trifluoroacetic acid gave the crys-
talline 4,6-O-benzylidene derivative 11 in 84% yield,
which was readily benzoylated to give the crystalline
ester 12 in 94% yield. Oxidative removal of the 4-
methoxyphenyl anomeric group in 12 with ceric ammo-
nium nitrate, followed by trichloroacetimidoylation
with trichloroacetonitrile and 1,8-diazabicyclo[5,4,0]-
undec-7-ene (DBU) gave the crystalline a-imidate 13 in
54% overall yield, the structure of which was evident
9.8
3.63
9.8
3.5
5.42
0.9
3.93
6.3
3.50
5.0
9.3
6.3
3.77
3.79
3.62
−10.2
3.59
1
from its H NMR spectrum (Table 2).
3.77
7.05
9.2
3.79
7.25
9.4
3.62
7.00
7.4
Preparation of the key intermediates 20 and 21 was
next studied (Scheme 4). Condensation of 7 and 8 with
a moderate excess (1.1 equiv) of 13, in dichloromethane
at 0 °C, in the presence of trimethylsilyl triflate, af-
forded readily the a-linked disaccharide derivatives 14
6.94
7.3
a Chemical shifts (l, ppm) and coupling constants (J, in
Hz) for solutions in CDCl3.

676
N. Barroca, J.-C. Jacquinet / Carbohydrate Research 337 (2002) 673–689
Scheme 3.
oxidant
and
sodium
hydrogencarbonate,
in
were then achieved as follows (Scheme 5). Condensa-
homogenous23 medium (acetonitrile–water), the corre-
sponding hydroxy-acid, sodium salts, thus obtained
were directly esterified with methyl iodide in N,N-
dimethylformamide at 50 °C to afford the methyl esters
16 and 18 in 63 and 66% overall yields, respectively.
tion of imidate 20 with the known10 methyl (methyl
2-O-benzoyl-3-O-benzyl-a- -idopyranosid)uronate (24)
L
(1.5 equiv), in dichloromethane at −15 °C, and in the
presence of trimethylsilyl triflate, gave the b-linked
1
trisaccharide derivative 25 in 61% yield. The H NMR
1
The H NMR spectra for 16 and 18 (Table 3) are in
spectrum for 25 (Table 4) shows a signal at l 4.86 ppm
with a large coupling constant (J1,2 8.4 Hz), in agree-
ment with a newly established 1,2-trans interglycosidic
linkage. According to the initial strategy, protection at
agreement with the expected structures, and show, for
the L-IdoA residues, long-range coupling constants (J1,3
1
and J2,4ꢀ1.0 Hz), indicating a high proportion of C4
conformation in solution. It is to be noted that such a
reaction works well on disaccharide units, and should
be applied for the synthesis of glycosaminoglycan disac-
charide derivatives. Conventional chloroacetylation at
O-4II on 16 and 18 gave the esters 17 and 19 in 85 and
91% yields, respectively, which were submitted to oxi-
dative cleavage of the 4-methoxyphenyl glycoside fol-
lowed by trichloroacetimidoylation, as reported above
for the preparation of 13, to give the key a-
trichloroacetimidates 20 and 21 in 55 and 58% overall
yields, respectively, the anomeries of which were de-
O-4 of the nonreducing L-IdoA residue by an orthogo-
nal (hydrogenolizable) protecting group was required.
To this end, ester 25 was O-dechloroacetylated to give
the corresponding alcohol, which is available for fur-
ther chain-extension at its nonreducing end. This inter-
mediate
was
treated
with
4-methoxyphenyl
trichloroacetimidate25 in dichloromethane, in the pres-
ence of trimethylsilyl triflate, to give the fully protected
Table 2
1H NMR data: carbohydrate ring protons for monosaccha-
1
ride derivatives 10–13 a
duced from their H NMR spectra (Table 3).
These versatile intermediates, ready to be used for
chain-extension at their reducing end, could be easily
transformed into a pair of glycosyl acceptors, ready for
further coupling at their nonreducing end. Methyl gly-
cosylation of 20 and 21 with methanol, in
dichloromethane at room temperature, and in the pres-
ence of trimethylsilyl triflate, provided the correspond-
Proton
10
11
12
13
H-1
J1,2
J1,3
H-2
J2,3
J2,4
5.71
1.1
1.1
5.47
2.2
1.1
5.64
1.0
0.9
4.10
2.6
1.0
5.76
1.1
1.0
5.49
2.3
0.9
6.54
1.0
1.0
5.43
2.6
0.9
ing methyl b-D-glycoside derivatives, which were
submitted to selective O-dechloroacetylation by treat-
ment with thiourea in pyridine and ethanol to give the
alcohols 22 and 23 in 77 and 79% overall yields,
respectively. In these glycosylation reactions, no methyl
H-3
J3,4
H-4
J4,5
H-5
J5,6a
J5,6b
H-6a
J6a,6b
H-6b
4.18
3.5
5.31
1.6
5.01
3.7
8.6
3.91
3.4
4.10
3.94
3.4
4.27
3.94
3.8
4.18
2.0
4.10
4.27
4.27
a-D-glycoside species were isolated, thus showing the
excellent stereocontrolling ability of the 2-deoxy-2-
trichloroacetamido group24 which is able to lead
stereoselectively to 1,2-trans glycosides with highly re-
active alcohol (i.e., methanol) as well as with low-reac-
tive hydroxy groups in sugar derivatives.17
4.64
4.27
−12.6
4.10
4.27
4.27
4.27
4.27
−11.7
4.44
a Chemical shifts (l, ppm) and coupling constants (J, in
Syntheses of the sulfated trisaccharide derivatives 1
and 2, having an
L
-IdoA residue at the reducing end,
Hz) for solutions in CDCl3.

N. Barroca, J.-C. Jacquinet / Carbohydrate Research 337 (2002) 673–689
677
methyl ester groups in 27 were removed by treatment
with lithium hydroperoxide26 in aqueous tetra-
hydrofuran to give the corresponding diacid derivative
without b-elimination side-reaction. Further in situ sa-
ponification with methanolic sodium hydroxide for 5 h,
followed by deionization with resin [H+ form] afforded
the pivaloylated hydroxy-acid 28 in 76% yield, whereas
the same treatment for 96 h gave the fully saponified
1
derivative 29 in 66% yield. The H NMR spectra for 28
and 29 (Table 4) show, apart from the presence in 28
and the absence in 29 of the pivalate ester (l 1.10 ppm,
s, 9 H), signals for H-4II at l 5.42 and 3.98 ppm,
respectively, in full agreement with the postulated struc-
tures. The intermediates 28 and 29 were then O-sul-
fonated by treatment with the sulfur trioxide–
trimethylamine complex in N,N-dimethylformamide at
50 °C, followed by ion-exchange chromatography [Na+
resin] to give the sodium salts 30 and 31 in 63 and 70%
1
yields, respectively. The H NMR spectra for 30 and 31
(Table 4) were compared with those of their non-sul-
fated precursors 28 and 29, respectively, all being
recorded in deuterated methanol under the same condi-
tions. Particularly relevant are the downfield shifts
(Dlꢀ0.5–0.6 ppm) of the signals for both IdoA H-2 in
30 and 31, and those (Dlꢀ0.8 ppm) of the signal for
GalNAc H-4 in 31. These chemical shift differences
indicate that sulfation occurred at O-2I and O-2III in
both derivatives, and at O-4II in 31, and are in complete
agreement with those reported for synthetic 2-O-sul-
fonated
4-O-sulfonated
L
-IdoA derivatives,9,10 as well as those for
D
-GalNAc residues in synthetic
oligosaccharides.17,27 Removal of the pivaloyl group in
sulfated 30 was achieved by treatment with methanolic
sodium hydroxide for 96 h to afford the disulfated
1
derivative 32 in 72% yield. The H NMR spectrum for
32 (Table 4), compared with those of its precursor 30,
shows a significant upfield shift (Dlꢀ1.6 ppm) for the
signal for GalNAc H-4 in 32 and no notable changes
for the other protons, in good agreement with the
expected structure. Final hydrogenation of 31 and 32
with Pd–C in aqueous methanol gave the target
molecule 1 and 2 in 93 and 96% yields, respectively.
1
The H (Table 4) and 13C NMR spectra for 1 and 2 are
in full agreement with the postulated structures, and in
accord with those reported28 for DS polymer isolated
from ascidians. The coupling constants observed for the
L
-IdoA residues indicate once more a predominance of
Scheme 4.
1C4 conformation in solution for both reducing and
nonreducing units.
trisaccharide derivative 26 in 68% overall yield. The
N-trichloroacetyl group in 26 was readily transformed
into N-acetyl by radical reduction24 with tributyltin
hydride and azobisisobutyronitrile (AIBN) to give the
crystalline acetamide 27 in 92% yield.
Synthesis of the sulfated trisaccharide derivative 3
having a
D-GalNAc residue at the reducing end was
then achieved as follows (Scheme 6). Condensation of
acceptors 22 and 23 with the known10 4-O-acetyl-
3,6-di-O-benzyl-2-deoxy-2-trichloroacetamido-1-O-tri-
Selective removal of the ester groups in 27 by con-
trolled saponification was next examined. Firstly, the
chloroacetimidoyl-a-
D
-galactopyranose (33) (1.5 equiv),
in dichloromethane at −15 °C, and in the presence of

678
N. Barroca, J.-C. Jacquinet / Carbohydrate Research 337 (2002) 673–689
1
trimethylsilyl triflate, afforded the trisaccharide deriva-
tives 34 and 35 in 80 and 75% yields, respectively, the
structures of which could be once more determined on
the basis of their 1H NMR data (Table 5). Radical
reduction on 34 and 35, as described above for the
preparation of 27, gave the acetamides 36 and 37 in 84
and 88% yields, respectively. Both compounds 36 and
37 were treated with lithium hydroperoxide in aqueous
tetrahydrofuran followed by methanolic sodium hy-
droxide for 5 h to give the pivaloylated derivative 38
and the fully saponified derivative 39, both in 95%
yield. Comparison of the H NMR spectra of 40 and
39, both recorded in deuterated methanol (Table 5),
showed the expected downfield shifts (Dlꢀ0.9 and 0.5
ppm) of the signals for GalNAc H-4 and IdoA H-2,
respectively. Final hydrogenation of 40 with Pd–C in
aqueous methanol gave the target molecule 3 in 88%
yield. The H (Table 5) and 13C NMR data for 3 are in
1
complete agreement with the expected structure, and fit
well with those reported for 1 and 2, as well as with
those reported28 for DS of natural origin.
To further test the efficiency of our synthetic route29
(not described in Section 3), the pivaloylated donor 20
(1.1 equiv) was condensed with the acetylated acceptor
23 (1 equiv) in dichloromethane at −15 °C, and in the
presence of trimethylsilyl triflate, to give the corre-
sponding tetrasaccharide derivative in 63% yield (l 4.85
ppm, d, 1 H, J1,2 8.4 Hz, H-1III). In this derivative, the
1
yields. The H NMR spectra for 38 and 39 (Table 5)
accord with the expected structures and with previous
observations reported above for 28 and 29. The mo-
noester 38 might be used for the preparation of the
corresponding disulfated species, showing that it should
be possible to differentiate two
D-GalNAc residues in
such a structure. It is to be noted that treatment of ester
38 with methanolic sodium hydroxide for 96 h gave the
hydroxy-acid 39 in 61% yield. O-Sulfonation of fully
saponified 39, as described above for the preparation of
sulfates 30 or 31, afforded the sodium salt 40 in 65%
two D-GalNAc residues might be once more differenti-
ated, starting from a common precursor.
In conclusion, an expeditious preparation of a pair of
key activated disaccharide intermediates bearing a vari-
ety of ester groups is reported. Stereocontrolled and
Table 3
1H NMR data: carbohydrate ring protons for disaccharide derivatives 14–23 a
Proton
14
5.38
8.4
3.95
11.0
4.58
3.2
5.58
0.5
15
5.41
8.5
3.90
11.0
4.61
3.4
5.57
0.5
16
17
18
5.29
8.5
4.13
10.8
4.56
3.1
5.55
0.5
19
5.37
8.4
3.90
10.9
4.65
3.2
5.52
0.5
20
21
22
4.78
8.5
3.74
11.1
4.49
3.0
5.53
0.5
23
H-1I
J1,2
5.27
8.4
4.05
11.5
4.51
2.9
5.29
8.4
4.03
10.9
4.51
3.1
6.46
3.7
4.83
6.42
3.8
4.80
10.8
4.22
3.0
4.80
8.4
3.76
10.9
4.56
3.2
H-2I
J2,3
10.8
H-3I
J3,4
4.22
3.3
5.65
0.7
H-4I
J4,5
5.54
0.5
5.53
0.5
5.60
0.9
5.55
0.5
3.92
5.6
6.6
3.54
H-5I
J5,6a
J5,6b
H-6aI
J6a,6b
H-6bI
NH
J2,NH
H-1II
J1,2
3.95
6.2
6.2
3.96
6.1
6.1
4.05
4.7
6.9
3.93
4.7
6.2
4.01
6.0
6.0
3.96
6.2
6.2
4.31
6.2
6.2
4.28
5.8
5.8
3.88
6.0
6.0
3.54
3.58
3.54
3.52
3.54
3.56
3.51
3.52
3.47
−10.1
3.48
−10.4
3.46
−10.2
3.43
−10.0
3.43
−10.4
3.49
3.54
6.90
7.6
5.19
2.8
3.58
6.88
7.3
5.19
2.3
3.54
6.95
7.8
5.23
1.1
3.56
6.95
7.8
5.22
1.1
3.47
6.03
7.6
5.18
1.2
7.07
7.8
5.21
1.0
7.02
7.8
5.24
1.0
6.82
9.8
5.31
1.1
6.72
9.6
5.35
1.1
6.96
7.8
5.18
1.2
J1,3
1.0
5.15
2.7
1.0
5.12
2.6
0.9
5.16
2.5
0.9
5.15
2.4
1.0
5.12
2.3
0.9
5.14
2.3
1.0
5.12
2.9
1.0
5.17
2.7
H-2II
J2,3
5.27
5.0
5.30
4.3
J2,4
1.0
3.75
2.6
4.05
1.8
4.81
1.0
3.82
2.6
5.27
2.1
4.93
0.9
3.81
2.5
4.13
1.9
5.07
1.0
3.88
2.6
4.34
2.0
5.12
1.0
3.84
2.7
5.28
1.8
4.98
1.1
3.91
2.1
5.32
1.5
5.19
1.2
3.82
2.5
4.04
1.8
4.81
1.2
3.79
2.6
4.11
1.8
5.02
H-3II
J3,4
3.80
2.8
4.05
3.82
2.6
4.18
H-4II
J4,5
H%%5II
H-6aII
H-6bII
4.05
4.45
4.15
4.18
4.45
4.18
a Chemical shifts (l, ppm) and coupling constants (J, in Hz) for solutions in CDCl3.

N. Barroca, J.-C. Jacquinet / Carbohydrate Research 337 (2002) 673–689
679
building block strategy used herein might be extended
for the construction of larger molecules. The sulfated
molecules 1–3 are currently being evaluated in biologi-
cal assays, and the results of these studies will be
reported elsewhere in due course.
3. Experimental
General methods.—Melting points were determined
in capillary tubes with a Bu¨chi apparatus and are
uncorrected. Optical rotations were measured at 20–
1
25 °C with a Perkin–Elmer 241 polarimeter. H NMR
spectra were recorded at 25 °C with Bruker DPX 250
and Varian Unity 500 spectrometers operating at 250
and 500 MHz, respectively, with Me4Si as internal
standard, unless otherwise stated. 13C NMR spectra
were recorded with a Bruker Advance DPX 250 operat-
ing at 62.8 MHz. Assignments were based on homo-
and heteronuclear correlations using the supplier’s soft-
ware. Low-resolution mass spectra were obtained on a
Perkin–Elmer SCIEX API 300 spectrometer operating
in the ion-spray (IS) mode. Flash-column chromatogra-
phy was performed on Silica Gel (E. Merck, 40–63
mm). Elemental analyses were performed by the Service
Central de Microanalyse du CNRS (Vernaison,
France).
4-Methoxyphenyl 4,6-O-benzylidene-2-deoxy-3-O-pi-
6aloyl-2-trichloroacetamido-i- -glucopyranoside (5).—
D
Pivaloyl chloride (7 mL, 57 mmol) was added dropwise
at 0 °C to a solution of 4-methoxyphenyl 4,6-O-benzyl-
idene-2-deoxy-2-trichloroacetamido-b-D-glucopyran-
oside (4)10 (9.9 g, 19 mmol) and DMAP (244 mg, 1.9
mmol) in pyridine (30 mL) and CH2Cl2 (70 mL), and
the mixture was stirred overnight at rt. Methanol (20
mL) was added, and the mixture was diluted with
CH2Cl2 (100 mL), washed with water, satd aq NaHCO3
and water, dried (MgSO4) and concentrated. The
residue was crystallized from EtOAc–petroleum ether
to give 5 (9.4 g, 82%); mp 205–206 °C; [h]D −65° (c 1,
CHCl3); 1H NMR (250 MHz, CDCl3): carbohydrate
ring protons (see Table 1); l 7.50–6.70 (m, 9 H,
aromatic H), 5.52 (s, 1 H, PhCH), 3.77 (s, 3 H, OCH3);
ISMS: m/z 621, [M+NH4]+ for 35Cl. Anal. Calcd for
C27H30Cl3NO8: C, 53.79; H, 5.01; N, 2.33. Found: C,
53.64; H, 5.00; N, 2.28.
Scheme 5.
high-yielding coupling reactions with moderate excess
of donors were achieved by using activated 2-deoxy-2-
trichloroacetamido-1-O-trichloroacetimidoyl-D-galacto
derivatives. Controlled saponification of ester groups
open the way for the preparation of various sulfoforms
starting from a common precursor. This strategy can be
applied not only for the synthesis of DS fragments, as
depicted herein, but also for the preparation of sulfo-
forms in chondroitin sulfates. In that case, which is
currently being investigated, no influence of the nature
4-Methoxyphenyl 6-O-benzyl-2-deoxy-3-O-pi6aloyl-
2-trichloroacetamido-i- -glucopyranoside (6).—Trifl-
D
uoroacetic acid (6 mL, 80 mmol) was added at 0 °C
under dry Ar to a solution of 5 (9.4 g, 15.5 mmol) and
triethylsilane (12.5 mL, 78 mmol) in anhyd CH2Cl2 (50
mL), and the mixture was stirred for 1 h at 0 °C and 2
h at rt. Triethylamine (30 mL) was added, and the
mixture was diluted with CH2Cl2 (100 mL), washed
with water, satd aq NaHCO3 and water, dried (MgSO4)
of the adjacent sugar residue (
IdoA) could be observed. Finally, the disaccharide
D-GlcA instead of L-

680
N. Barroca, J.-C. Jacquinet / Carbohydrate Research 337 (2002) 673–689
dropwise at −15 °C under dry Ar to a solution of 6
(8.6 g, 14.2 mmol) in anhyd pyridine (5.8 mL) and
anhyd 1,2-dichloroethane (50 mL), and the mixture was
stirred for 90 min at this temperature. Water (6.4 mL)
was added, and the mixture was stirred for 30 min at
80 °C, then was cooled, diluted with CH2Cl2 (50 mL),
washed with water, satd aq NaHCO3 and water, dried
(MgSO4) and concentrated. The residue was eluted
from a column (100 g) of silica gel with 3:2 petroleum
ether–EtOAc to give 7 as a white foam (7.0 g, 82%);
and concentrated. The residue was eluted from a
column (100 g) of silica gel with 20:1 CH2Cl2–EtOAc
to afford 6 as a white foam (8.6 g, 91%); [h]D −32° (c
1
1, CHCl3); H NMR (250 MHz, CDCl3): carbohydrate
ring protons (see Table 1); l 7.36–6.80 (m, 9 H,
aromatic H), 4.57 (ABq, 2 H, CH2Ph), 3.75 (s, 3 H,
OCH3), 3.00 (d, 1 H, J 4.7 Hz, HO-4), 1.20 (s, 9 H,
((CH3)3C); ISMS: m/z 628, [M+Na]+ for 35Cl. Anal.
Calcd for C27H32Cl3NO8: C, 53.61; H, 5.33; N, 2.33.
Found: C, 53.69; H, 5.13; N, 2.41.
1
[h]D −34° (c 1, CHCl3); H NMR (250 MHz, CDCl3):
4-Methoxyphenyl 6-O-benzyl-2-deoxy-4-O-pi6aloyl-
carbohydrate ring protons (see Table 1); l 7.40–6.90
2-trichloroacetamido-i-
D
-galactopyranoside
(7).—
(m, 9 H, aromatic H), 4.51 (ABq, 2 H, CH2Ph), 3.76 (s,
Triflic anhydride (2.8 mL, 17 mmol) was added
Table 4
1H NMR data: carbohydrate ring protons for trisaccharide derivatives 25–32, 1 and 2 a
b
Proton
25
4.91
1.2
0.9
26
4.90
1.1
1.0
27
28
29 b
30 b
31 b
32 b
1 c
4.83
2 c
4.94
1.0
1.0
4.07
2.2
H-1I
J1,2
4.95
1.0
4.85
4.78
5.15
1.0
5.03
1.0
5.12
1.0
1.0
1.0
4.09
2.0
1.0
1.0
4.05
2.1
1.0
J1,3
0.9
H-2I
J2,3
5.07
2.4
5.13
2.3
5.10
2.0
4.66
2.0
4.62
4.62
4.06 d
2.5
J2,4
1.2
4.09
2.5
4.14
2.0
4.83
4.86
8.4
3.76
10.8
4.19
2.8
5.38
0.5
1.0
4.10
2.6
4.15
2.6
4.68
4.88
8.4
3.80
11.0
4.20
3.0
5.37
0.5
H-3I
J3,4
4.20
2.2
4.15
1.5
3.95
4.05
4.13
4.12
4.10
4.10
4.31
2.2
4.07
1.5
4.19
2.0
3.98
1.5
H-4I
J4,5
3.96
2.0
4.05
1.5
4.24
1.6
4.15
1.5
H-5I
H-1II
J1,2
4.63
4.88
8.2
4.56
4.62
8.4
4.48
4.56
8.4
4.53
4.68
8.2
4.44
4.62
8.4
4.42
4.64
8.4
4.68
4.68
8.4
4.54
4.64
8.4
H-2II
J2,3
3.20
9.9
3.90
10.0
3.80
10.0
3.90
10.0
3.95
10.0
3.80
3.70
10.0
3.80
10.0
H-3II
J3,4
4.38
2.9
3.68
3.1
3.65
3.0
3.58
3.0
3.82
3.0
3.80
3.0
3.94
3.0
3.85
3.0
H-4II
J4,5
5.33
0.5
5.42
0.5
3.98
0.5
5.58
0.5
4.84
0.5
3.98
0.5
4.74
0.5
3.96
0.5
H-5II
H-6aII
H-6bII
NH
3.80
3.23
3.23
6.79
7.9
3.70
3.28
3.24
6.84
7.9
3.70
3.21
3.21
5.62
7.0
3.65
3.55
3.55
3.93
3.65
3.65
3.90
3.55
3.55
3.82
3.75
3.75
3.80
3.70
3.70
3.88
3.70
3.70
3.82
3.68
3.68
J2,NH
H-1III
J1,2
5.20
1.0
5.22
1.9
5.03
1.2
5.01
3.0
5.05
3.2
5.34
1.2
5.27
1.2
5.42
1.2
5.11
1.0
5.16
1.0
J1,3
0.8
5.14
2.0
1.0
4.64
2.0
1.0
4.58
1.0
4.18
2.5
1.0
4.18
2.3
H-2III
J2,3
5.05
3.4
5.15
2.9
4.08
2.5
4.05
4.62
J2,4
1.0
3.76
2.2
5.23
1.2
4.80
1.2
3.76
2.9
3.80
1.8
4.81
H-3III
J3,4
3.75
3.95
4.05
3.75
4.84
3.98
4.12
4.10
3.70
4.38
4.32
2.6
3.96
1.5
4.35
2.5
3.94
1.5
H-4III
J4,5
3.80
2.3
4.80
3.65
2.5
4.66
3.60
1.7
4.57
3.72
1.8
4.34
H-5III
4.74
4.44
a For solutions in CDCl3, unless otherwise stated.
b CD3OD.
c D2O.
d Values in bold type reflect the locations of the sulfate groups.

N. Barroca, J.-C. Jacquinet / Carbohydrate Research 337 (2002) 673–689
681
mL), then was deionized with Amberlite IR–120 [H+]
resin, filtered and concentrated to afford the corre-
sponding diol (6.2 g, quantitative), mp 172–173 °C
(from EtOAc–petroleum ether).
A mixture of the above isolated diol (6.2 g, 11.8
mmol), trimethyl orthoacetate (17.5 mL, 140 mmol)
and 10-D,L-camphorsulfonic acid (0.46 g) in anhyd
toluene (30 mL) was stirred for 1 h at rt. Triethylamine
(0.83 mL) was added, and the mixture was diluted with
toluene (30 mL), washed with water, dried (MgSO4)
and concentrated. A solution of the residue in 80%
AcOH (100 mL) was stirred for 10 min at rt, then
concentrated, evaporated with water and toluene. The
residue was crystallized from EtOAc–petroleum ether
to give 8 (5.7 g, 86% from 7); mp 146–147 °C; [h]D
−34° (c 1, CHCl3); 1H NMR (250 MHz, CDCl3):
carbohydrate ring protons (see Table 1); l 7.40–6.80
(m, 9 H, aromatic H), 4.48 (ABq, 2 H, CH2Ph), 3.76 (s,
3 H, OCH3), 2.89 (d, 1 H, J 5.2 Hz, HO-3), 2.12 (s, 3
H, Ac); 13C NMR (CDCl3): selected data; l 99.92
(C-1), 92.46 (CCl3), 73.79 (CH2Ph), 72.95 (C-5), 69.82
(C-4), 69.32 (C-3), 68.21 (C-6), 56.65 (C-2), 55.74
(OCH3), 20.95 (COCH3); ISMS: m/z 581, [M+NH4]+
for 35Cl. Anal. Calcd for C24H26Cl3NO8: C, 51.21; H,
4.66; N, 2.50. Found: C, 51.19; H, 4.67; N, 2.50.
4-Methoxyphenyl 2,4,6-tri-O-benzoyl-3-O-benzyl-h-
L
-idopyranoside (10).—A mixture of 1,2,4,6-tetra-O-
benzoyl-3-O-benzyl-b-
L
-idopyranose (9)10 (7.70 g, 11.2
,
mmol), 4-methoxyphenol (2.80 g, 22.6 mmol), and 4 A
powdered molecular sieves (5 g) in anhyd CH2Cl2 (40
mL) was stirred for 1 h at rt under dry Ar. Trimethylsi-
lyl triflate (2.0 mL, 11.2 mmol) was added, and the
mixture was stirred for 30 min at rt. Triethylamine (4.6
mL) was then added, and the mixture was diluted with
CH2Cl2 (30 mL), filtered through a pad of Celite,
washed with water, 1 M NaOH and water, dried
(MgSO4) and concentrated. The residue was eluted
from a column (80 g) of silica gel with 4:1 petroleum
ether–EtOAc to give 10 as a light yellow oil (7.70 g,
1
98%); [h]D −42° (c 1, CHCl3); H NMR (250 MHz,
CDCl3): carbohydrate ring protons (see Table 2); l
8.20–6.80 (m, 24 H, aromatic H), 4.94 (ABq, 2 H,
CH2Ph), 3.70 (s, 3 H, OCH3); ISMS: m/z 706, [M+
NH4]+, 565, [M−OC6H4OCH3]+. Anal. Calcd for
C41H36O10: C, 71.50; H, 5.27. Found: C, 71.62; H, 5.13.
Scheme 6.
3 H, OCH3), 2.76 (d, 1 H, J 4.6 Hz, HO-3), 1.20 (s, 9
H, (CH3)3C); 13C NMR (CDCl3): selected data; l 99.67
(C-1), 92.48 (CCl3), 73.82 (CH2Ph), 73.12 (C-5), 69.68
(C-3), 69.43 (C-4), 68.34 (C-6), 56.67 (C-2), 55.75
(OCH3), 39.45 ((CH3)3C), 27.35 ((CH3)3C); ISMS: m/z
628, [M+Na]+ for 35Cl. Anal. Calcd for
C27H32Cl3NO8: C, 53.61; H, 5.33; N, 2.33. Found: C,
53.71; H, 5.37; N, 2.33.
4-Methoxyphenyl 3-O-benzyl-4,6-O-benzylidene-h-L-
idopyranoside (11).—A solution of 10 (7.70 g, 11.2
mmol) in MeOH (100 mL) was transesterified as de-
scribed for the preparation of 8. A mixture of the
residue, benzaldehyde (20 mL) and TFA (1 mL) was
stirred for 3 h at rt. Triethylamine (6.5 mL) and CH2Cl2
(100 mL) were then added, and the mixture was washed
with brine and water, dried (MgSO4) and concentrated.
The residue was eluted from a column (100 g) of silica
gel with 10:1“4:1 petroleum ether–EtOAc, and crys-
tallized from EtOAc–petroleum ether to give 11 (4.37
4-Methoxyphenyl 4-O-acetyl-6-O-benzyl-2-deoxy-2-
trichloroacetamido-i- -galactopyranoside (8).—A solu-
D
tion of 7 (7.1 g, 11.8 mmol) in MeOH (100 mL) was
treated for 3 h at rt with methanolic MeONa (1 M, 3

682
N. Barroca, J.-C. Jacquinet / Carbohydrate Research 337 (2002) 673–689
(100 mL), and the mixture was stirred for 1 h at this
temperature. Methanol (10 mL) was added, and the
mixture was diluted with CH2Cl2 (100 mL), washed
with water, satd aq NaHCO3 and water, dried (MgSO4)
and concentrated. The residue was crystallized from
EtOAc–petroleum ether to give 12 (12.30 g, 94%); mp
g, 84% from 10); mp 121–122 °C; [h]D −70° (c 1,
CHCl3); 1H NMR (250 MHz, CDCl3): carbohydrate
ring protons (see Table 2); l 7.50–7.0 (m, 14 H,
aromatic H), 5.55 (s, 1 H, PhCH), 4.76 (ABq, 2 H,
CH2Ph), 3.80 (s, 3 H, OCH3), 3.46 (d, 1 H, J 11.5 Hz,
HO-2); ISMS: m/z 591, [M+Na]+. Anal. Calcd for
C34H32O8: C, 71.82; H, 5.67. Found: C, 71.78; H, 5.53.
1
127–128 °C; [h]D −54° (c 1, CHCl3); H NMR (250
MHz, CDCl3): carbohydrate ring protons (see Table 2);
l 8.0–6.90 (m, 19 H, aromatic H), 5.60 (s, 1 H, PhCH),
4.87 (ABq, 2 H, CH2Ph), 3.70 (s, 3 H, OCH3); ISMS:
m/z 591, [M+Na]+, 445, [M−OC6H4OCH3]+. Anal.
4-Methoxyphenyl
2-O-benzoyl-3-O-benzyl-4,6-O-
benzylidene-h- -idopyranoside (12).—Benzoyl chloride
L
(5.4 mL, 46.5 mmol) was added at 0 °C to a solution of
11 (10.70 g, 23 mmol) in pyridine (19 mL) and CH2Cl2
Table 5
1H NMR data: carbohydrate ring protons for trisaccharide derivatives 34–40, and 3 a
b
Proton
34
35
36
37
38
39 b
4.44
8.4
3.92
11.0
4.24
3.0
4.05
0.5
3.65
3.65
3.65
40 b
4.55
8.2
3.90
11.0
4.12
3.0
4.96
0.5
3.65
3.65
3.65
3 c
4.55
8.0
3.92
H-1I
J1,2
4.77
8.2
3.74
10.8
4.46
2.9
5.51
0.5
3.86
3.50
3.50
6.88
7.6
4.81
8.4
3.76
11.3
4.30
3.1
5.50
0.5
3.85
3.52
3.52
6.98
7.6
4.81
8.5
3.55
11.2
4.41
2.9
5.50
0.5
3.87
3.45
3.45
5.81
7.9
4.94
8.4
3.54
11.0
4.40
2.9
5.52
0.5
3.84
3.46
3.46
5.72
7.9
4.52
8.2
3.93
11.0
4.17
2.9
5.43
0.5
3.65
3.50
3.50
H-2I
J2,3
10.5
H-3I
J3,4
3.75
3.0
H-4I
J4,5
4.73 d
0.5
H-5I
H-6aI
H-6bI
NH
3.70
3.70
3.70
J2,NH
H-1II
J1,2
5.12
1.0
5.14
1.0
5.09
1.0
5.10
1.0
5.03
1.0
5.28
1.0
5.36
1.8
5.07
1.0
J1,3
1.0
4.99
2.1
1.0
5.07
2.2
1.0
4.97
2.3
1.0
5.03
2.2
1.0
4.10
0.9
4.12
0.9
4.58
2.5
1.0
4.06
2.5
H-2II
J2,3
J2,4
0.9
4.01
2.5
4.13
1.8
4.76
4.80
8.4
3.50
11.1
3.97
3.3
5.46
0.5
0.9
4.08
2.3
4.20
1.5
4.94
4.85
8.4
3.52
10.7
3.93
3.1
5.47
0.5
0.9
4.01
2.5
4.05
1.6
4.74
4.91
8.2
3.16
11.3
4.36
2.8
5.42
0.5
0.8
4.09
2.4
4.09
2.0
4.92
4.98
8.4
3.46
11.0
4.33
3.0
5.40
0.5
1.0
3.97
2.2
4.12
1.5
4.66
4.53
8.2
3.92
10.5
3.75
3.0
4.56
0.5
H-3II
J3,4
3.92
4.05
4.28
2.5
4.38
1.6
4.95
4.52
8.3
3.90
11.0
3.65
3.0
4.94
0.5
H-4II
J4,5
4.06
2.4
4.87
4.48
8.4
3.95
11.0
3.65
3.5
4.02
0.5
4.18
1.6
4.92
4.42
8.2
3.92
11.0
3.65
3.5
4.05
0.5
H-5II
H-1III
J1,2
H-2III
J2,3
H-3III
J3,4
H-4III
J4,5
H-5III
H-6aIII
H-6bIII
NH
3.55
3.00
3.00
6.74
7.6
3.62
3.14
3.14
6.75
7.8
3.65
3.15
3.15
5.65
7.2
3.73
3.27
3.27
5.58
6.6
3.65
3.45
3.45
3.65
3.50
3.50
3.65
3.50
3.50
3.70
3.66
3.66
J2,NH
a For solutions in CDCl3, unless otherwise stated.
b CD3OD.
c D2O.
d Values in bold type reflect the locations of the sulfate groups.

N. Barroca, J.-C. Jacquinet / Carbohydrate Research 337 (2002) 673–689
683
Calcd for C34H32O8: C, 71.82; H, 5.67. Found: C, 71.78;
H, 5.53.
C54H56Cl3NO14: C, 61.80; H, 5.38; N, 1.33. Found: C,
61.76; H, 5.28; N, 1.51.
4-Methoxyphenyl O-(2-O-benzoyl-3-O-benzyl-4,6-O-
2-O-Benzoyl-3-O-benzyl-4,6-O-benzylidene-1-O-tri-
benzylidene-h-
benzyl-2-deoxy-2-trichloroacetamido-i-
oside (15).—A mixture of alcohol 8 (5.80 g, 10.4 mmol)
and imidate 13 (6.90 g, 11.4 mmol) was treated as
described for the preparation of 14 to give 15 as a white
L
-idopyranosyl)-(1“3)-4-O-acetyl-6-O-
-galactopyran-
chloroacetimidoyl-h- -idopyranose (13).—A mixture of
L
D
12 (6.10 g, 10.8 mmol) and ceric ammonium nitrate
(29.7 g, 54.1 mmol) in 1:1.5:1 toluene–MeCN–water
(210 mL) was stirred vigorously for 5 min at rt, then
was diluted with EtOAc (300 mL), washed with water,
satd aq NaHCO3 and water, dried (MgSO4) and con-
centrated. The residue was eluted from a column (200
g) of silica gel with 5:2 petroleum ether–EtOAc to give
the corresponding hemiacetal (4.30 g).
1
foam (8.20 g, 79%); [h]D −0.5° (c 1, CHCl3); H NMR
(250 MHz, CDCl3): carbohydrate ring protons (see
Table 3); l 8.0–6.90 (m, 24 H, aromatic H), 5.58 (s, 1
H, PhCH), 4.73 (ABq, 2 H, CH2Ph), 4.51 (s, 2 H,
CH2Ph), 3.74 (s, 3 H, OCH3), 1.90 (s, 3 H, Ac); 13C
NMR (CDCl3): selected data; l 101.25 (C-1II), 100.75
(PhCH), 99.24 (C-1I), 92.19 (CCl3), 75.05 (C-3I), 74.87
(C-3II), 74.33 (C-5II), 73.69, 72.16 (CH2Ph), 73.09 (C-
5I), 69.61 (C-6II), 69.55 (C-4I), 68.48 (C-6I), 67.27 (C-
2II), 60.78 (C-4II), 55.86 (C-2I), 55.64 (OCH3), 20.60
(COCH3); ISMS: m/z 1024, [M+NH4]+ for 35Cl.
Anal. Calcd for C51H50Cl3NO14: C, 60.81; H, 5.00; N,
1.40. Found: C, 61.01; H, 5.15; N, 1.53.
A mixture of the above isolated hemiacetal, CCl3CN
(7.4 mL, 93 mmol), and DBU (0.6 mL, 4.7 mmol) in
anhyd CH2Cl2 (60 mL) was stirred for 1 h from 0 °C to
rt, then was concentrated. The residue was eluted from
a column (200 g) of silica gel with 4:1 petroleum
ether–EtOAc containing 0.2% of Et3N, and crystallized
from Et2O–petroleum ether to give 13 (3.50 g, 54%
1
from 12); mp 151–152 °C; [h]D −59° (c 1, CHCl3); H
NMR (250 MHz, CDCl3): carbohydrate ring protons
(see Table 2); l 8.70 (s, 1 H, CꢀNH), 8.0–7.20 (m, 15
H, Ph), 5.58 (s, 1 H, PhCH), 4.85 (ABq, 2 H, CH2Ph);
13C NMR (CDCl3): selected data; l 165.62 (CꢀO),
160.40 (CꢀNH), 137.91–125.51 (aromatic C), 101.17
(PhCH), 95.82 (C-1), 91.26 (CCl3), 73.23 (C-3), 72.77
(C-5), 72.14 (CH2Ph), 69.50 (C-6), 64.30 (C-2), 61.86
(C-4); ISMS: m/z 445, [M−OC(NH)CCl3]+ for 35Cl.
Anal. Calcd for C29H26Cl3NO7: C, 57.39; H, 4.32; N,
2.32. Found: C, 57.43; H, 4.26; N, 2.35.
4-Methoxyphenyl O-(methyl 2-O-benzoyl-3-O-benzyl-
h-
L
-idopyranosyluronate)-(1“3)-6-O-benzyl-2-deoxy-
-galactopyranoside
4-O-pi6aloyl-2-trichloroacetamido-i-
D
(16).—A mixture of 14 (3.57 g, 3.4 mmol) and AcOH
(25 mL) was stirred at 100 °C. Water (15 mL) was
added dropwise, and the mixture was stirred for 30 min
at 100 °C, then was cooled, concentrated, evaporated
with water and toluene to give the corresponding diol
(3.20 g).
A solution of calcium hypochlorite (1.50 g, 10.4
mmol) and NaHCO3 (0.87 g, 10.4 mmol) in water (45
mL) was added dropwise at 0 °C to a solution of the
above isolated diol, 1.6×10−2 M TEMPO in CH2Cl2
(10.8 mL, 0.17 mmol) and 0.5 M aq KBr (0.69 mL) in
MeCN (45 mL), and the mixture was stirred for 45 min
at 0 °C, then was concentrated and evaporated with
toluene.
Methyl iodide (0.96 mL, 15.5 mmol) was added to a
solution of the residue in DMF (50 mL), and the
mixture was stirred for 1 h at 50 °C, then was cooled,
poured into ice-cold 5% aq HCl and extracted with
EtOAc (3×100 mL). The organic extracts were washed
with 5% aq Na2S2O3, brine and water, dried (MgSO4)
and concentrated. The residue was eluted from a
column (150 g) of silica gel with 5:1 CH2Cl2–EtOAc to
give 16 as a white foam (2.20 g, 63% from 14); [h]D
−8° (c 1, CHCl3); 1H NMR (250 MHz, CDCl3): carbo-
hydrate ring protons (see Table 3); l 8.0–6.86 (m, 19
H, aromatic H), 4.69 (ABq, 2 H, CH2Ph), 4.47 (s, 2 H,
CH2Ph), 3.80 (s, 3 H, COOCH3), 3.69 (s, 3 H, OCH3),
2.89 (d, 1 H, J 10.8 Hz, HO-4II), 1.19 (s, 9 H, (CH3)3C);
ISMS: m/z 1007, [M+NH4]+ for 35Cl. Anal. Calcd for
C48H52Cl3NO15: C, 58.27; H, 5.30; N, 1.41. Found: C,
58.07; H, 5.38; N, 1.55.
4-Methoxyphenyl O-(2-O-benzoyl-3-O-benzyl-4,6-O-
benzylidene-h-
deoxy-4-O-pi6aloyl-2-trichloroacetamido-i-
pyranoside (14).—A mixture of alcohol 7 (3.20 g, 7.3
L
-idopyranosyl)-(1“3)-6-O-benzyl-2-
D
-galacto-
,
mmol), imidate 13 (3.40 g, 5.7 mmol), and 4 A pow-
dered molecular sieves (5 g) in anhyd CH2Cl2 (50 mL)
was stirred for 1 h at rt under dry Ar, then cooled to
0 °C. A solution of Me3SiOTf in toluene (1 M, 0.86
mL) was added, and the mixture was stirred for 30 min
at 0 °C. Triethylamine (0.36 mL) was added, and the
mixture was filtered and concentrated. The residue was
eluted from a column (200 g) of silica gel with 4:1
petroleum ether–EtOAc containing 0.2% of Et3N to
give 14 as a white foam (4.30 g, 78%); [h]D −10° (c 1,
CHCl3); 1H NMR (250 MHz, CDCl3): carbohydrate
ring protons (see Table 3); l 8.0–6.80 (m, 24 H,
aromatic H), 5.56 (s, 1 H, PhCH), 4.70 (ABq, 2 H,
CH2Ph), 4.50 (s, 2 H, CH2Ph), 3.74 (s, 3 H, OCH3),
1.20 (s, 9 H, (CH3)3C); 13C NMR (CDCl3): selected
data; l 101.83 (C-1II), 100.80 (PhCH), 99.06 (C-1I),
92.17 (CCl3), 75.97 (C-3I), 75.28 (C-5II), 75.21 (C-3II),
73.73, 72.11 (CH2Ph), 73.41 (C-5I), 69.66 (C-6II), 69.33
(C-4I), 68.52 (C-6I), 67.95 (C-2II), 61.16 (C-4II), 55.89
(C-2I), 55.71 (OCH3), 39.29 ((CH3)3C), 27.34
((CH3)3C); ISMS: m/z 1071, [M+Na]+, 925, [M−
4-Methoxyphenyl O-(methyl 2-O-benzoyl-3-O-benzyl-
4-O-chloroacetyl-h-L-idopyranosyluronate)-(1“3)-6-O-
OC6H4OCH3]+
for
35Cl.
Anal.
Calcd
for

684
N. Barroca, J.-C. Jacquinet / Carbohydrate Research 337 (2002) 673–689
benzyl-2-deoxy-4-O-pi6aloyl-2-trichloroacetamido-i-
D
-
The residue was eluted from a column (100 g) of silica
gel with 6:1 petroleum ether–EtOAc to give the corre-
sponding free hemiacetal (1.41 g).
galactopyranoside (17).—Chloroacetic anhydride (0.75
g, 4.4 mmol) was added at 0 °C to a solution of 16 (2.20
g, 2.2 mmol) in anhyd pyridine (1.2 mL) and anhyd
CH2Cl2 (30 mL), and the mixture was stirred for 1 h at
0 °C. Crushed ice was added, and the mixture was
diluted with CH2Cl2 (50 mL), washed with water, satd
aq NaHCO3 and water, dried (MgSO4) and concen-
trated. The residue was eluted from a column (80 g) of
silica gel with 10:1 CH2Cl2–EtOAc to afford 17 as a
A mixture of the above isolated hemiacetal, CCl3CN
(1.3 mL, 13 mmol) and DBU (50 mL, 0.33 mmol) in
anhyd CH2Cl2 (30 mL) was stirred for 30 min at 0 °C,
then was concentrated. The residue was eluted from a
column (100 g) of silica gel with 5:2 petroleum ether–
EtOAc containing 0.2% of Et3N to give 20 as a white
foam (1.22 g, 55% from 17); [h]D +47° (c 1, CHCl3);
1H NMR (500 MHz, CDCl3): carbohydrate ring pro-
tons (see Table 3); l 8.80 (s, 1 H, CꢀNH), 8.0–7.20 (m,
15 H, Ph), 4.74, 4.45 (2 ABq, 4 H, CH2Ph), 3.84 (ABq,
2 H, CH2Cl), 3.80 (s, 3 H, COOCH3), 1.15 (s, 9 H,
(CH3)3C); 13C NMR (CDCl3): selected data; l 101.85
(C-1II), 95.38 (C-1I), 92.31, 90.79 (CCl3), 77.59 (C-3I),
73.54, 72.48 (CH2Ph), 71.76 (C-5I), 71.37 (C-3II), 69.54
(C-4II), 68.61 (C-4I), 67.99 (C-6I), 66.82 (C-2II), 66.46
(C-5II), 52.75 (COOCH3), 50.63 (C-2I), 40.38 (CH2Cl),
39.18 ((CH3)3C), 27.06 ((CH3)3C); ISMS: m/z 942,
[M−OC(NH)CCl3]+ for 35Cl. Anal. Calcd for
C45H47Cl7N2O15: C, 48.95; H, 4.29; N, 2.53. Found: C,
48.75; H, 4.12; N, 2.39.
1
white foam (2.0 g, 85%); [h]D +6° (c 1, CHCl3); H
NMR (250 MHz, CDCl3): carbohydrate ring protons
(see Table 3); l 8.0–6.80 (m, 19 H, aromatic H), 4.74,
4.47 (2 ABq, 4 H, CH2Ph), 3.83 (ABq, 2 H, CH2Cl),
3.77 (s, 3 H, COOCH3), 3.72 (s, 3 H, OCH3), 1.15 (s, 9
H, (CH3)3C); ISMS: m/z 1083, [M+NH4]+ for 35Cl.
Anal. Calcd for C50H53Cl4NO16: C, 56.35; H, 5.01; N,
1.32. Found: C, 56.49; H, 5.09; N, 1.42.
4-Methoxyphenyl O-(methyl 2-O-benzoyl-3-O-benzyl-
h -
L
- idopyranosyluronate) - (1“3) - 4 - O - acetyl - 6 - O-
-galactopyran-
benzyl-2-deoxy-2-trichloroacetamido-i-
D
oside (18).—Compound 15 (6.12 g, 6.1 mmol) was
submitted to acid hydrolysis, TEMPO oxidation, and
methyl esterification as described for the preparation of
16 to give 18 as a white foam (3.80 g, 66% from 15);
(Methyl 2-O-benzoyl-3-O-benzyl-4-O-chloroacetyl-h-
L
-idopyranosyluronate)-(1“3)-4-O-acetyl-6-O-benzyl-
2-deoxy-2-trichloroacetamido-1-O-trichloroacetimidoyl-
h- -galactopyranose (21).—Compound 19 (3.80 g, 3.7
1
[h]D −7° (c 1, CHCl3); H NMR (250 MHz, CDCl3):
carbohydrate ring protons (see Table 3); l 8.0–6.80 (m,
19 H aromatic H), 4.71, 4.46 (2 ABq, 4 H, CH2Ph),
3.79 (s, 3 H, COOCH3), 3.68 (s, 3 H, OCH3), 2.89 (d, 1
H, J 10.0 Hz, HO-4II), 1.82 (s, 3 H, Ac); ISMS: m/z
969, [M+Na]+ for 35Cl. Anal. Calcd for
C45H46Cl3NO15: C, 57.06; H, 4.89; N, 1.48. Found: C,
56.94; H, 4.94; N, 1.56.
D
mmol) was treated as described for the preparation of
20 to give 21 as a white foam (2.29 g, 58% from 19);
1
[h]D +51° (c 1, CHCl3); H NMR (500 MHz, CDCl3):
carbohydrate ring protons (see Table 3); l 8.83 (s, 1 H,
CꢀNH), 8.0–7.20 (m, 15 H, Ph), 4.76, 4.44 (2 ABq, 4
H, CH2Ph), 3.80 (ABq, 2 H, CH2Cl), 3.78 (s, 3 H,
COOCH3), 1.65 (s, 3 H, Ac); 13C NMR (CDCl3):
selected data; l 101.08 (C-1II), 95.20 (C-1I), 92.24, 90.69
(CCl3), 75.57 (C-3I), 73.57, 72.64 (CH2Ph), 71.41 (C-
3II), 71.19 (C-5I), 69.39 (C-4II), 68.54 (C-4I), 67.75 (C-
6I), 66.40 (C-5II), 66.28 (C-2II), 52.79 (COOCH3), 50.87
(C-2I), 40.20 (CH2Cl), 19.99 (COCH3); ISMS: m/z 900,
[M−OC(NH)CCl3]+ for 35Cl. Anal. Calcd for
C42H41Cl7NO15: C, 47.50; H, 3.89; N, 2.64. Found: C,
47.25; H, 3.93; N, 2.82.
4-Methoxyphenyl O-(methyl 2-O-benzoyl-3-O-benzyl-
4-O-chloroacetyl-h-
L
-idopyranosyluronate)-(1“3)-4-
O-acetyl-6-O-benzyl-2-deoxy-2-trichloroacetamido-i-
D
-
galactopyranoside (19).—Compound 18 (3.80 g, 4
mmol) was treated as described for the preparation of
17 to give 19 as a white foam (3.80 g, 91%); [h]D +6°
1
(c 1, CHCl3); H NMR (250 MHz, CDCl3): carbohy-
drate ring protons (see Table 3); l 8.0–6.80 (m, 19 H
aromatic H), 4.77, 4.49 (2 ABq, 4 H, CH2Ph), 3.87
(ABq, 2 H, CH2Cl), 3.80 (s, 3 H, COOCH3), 3.74 (s, 3
H, OCH3), 1.82 (s, 3 H, Ac); ISMS: m/z 1039, [M+
NH4]+ for 35Cl. Anal. Calcd for C47H47Cl4NO16: C,
55.14; H, 4.63; N, 1.37. Found: C, 55.01; H, 4.74; N,
1.42.
Methyl
idopyranosyluronate)-(1“3)-6-O-benzyl-2-deoxy-4-O-
pi6aloyl - 2 - trichloroacetamido - i - - galactopyranoside
(22).—A mixture of imidate 20 (1.10 g, 0.98 mmol),
O-(methyl
2-O-benzoyl-3-O-benzyl-h-L-
D
,
anhyd MeOH (0.2 mL, 4.9 mmol) and 3 A powdered
(Methyl 2-O-benzoyl-3-O-benzyl-4-O-chloroacetyl-h-
molecular sieves (1 g) in anhyd CH2Cl2 (20 mL) was
stirred for 1 h at rt under dry Ar. A solution of
Me3SiOTf in toluene (1 M, 0.15 mL) was added, and
the mixture was stirred for 30 min. Triethylamine (0.1
mL) was added, and the mixture was filtered and
concentrated. The residue was eluted from a column
(75 g) of silica gel with 2:1 petroleum ether–EtOAc
containing 0.2% of Et3N to give the corresponding
methyl glycoside (772 mg, 81%).
L
-idopyranosyluronate)-(1“3)-6-O-benzyl-2-deoxy-4-
O-pi6aloyl-2 -trichloroacetamido-1 -O-trichloroacetimi-
doyl-h- -galactopyranose (20).—A mixture of 17 (2.10
D
g, 2 mmol) and ceric ammonium nitrate (8.30 g, 15
mmol) in 1:1.5:1 toluene–MeCN–water (52.5 mL) was
stirred vigorously for 15 min at rt, then was diluted
with EtOAc (100 mL), washed with water, satd aq
NaHCO3 and water, dried (MgSO4) and concentrated.

N. Barroca, J.-C. Jacquinet / Carbohydrate Research 337 (2002) 673–689
685
A mixture of the methyl glycoside and thiourea (0.25
g, 3.2 mmol) in pyridine (3.5 mL) and abs EtOH (7 mL)
was stirred for 30 min at 80 °C, then was cooled and
concentrated. A solution of the residue in CH2Cl2 (30
mL) was washed with water, satd aq NaHCO3 and
water, dried (MgSO4) and concentrated. The residue
was eluted from a column (75 g) of silica gel with 1:1
EtOAc–petroleum ether to give 22 as a white foam
foam (818 mg, 61%); [h]D −9.5° (c 1, CHCl3); 1H
NMR (500 MHz, CDCl3): carbohydrate ring protons
(see Table 4); l 8.10–7.20 (m, 25 H, Ph), 4.74, 4.70,
4.37 (3 ABq, 6 H, CH2Ph), 3.82, 3.75 (2 s, 6 H,
COOCH3), 3.78 (ABq, 2 H, CH2Cl), 3.46 (s, 3 H,
OCH3), 0.82 (s, 9 H, (CH3)3C); 13C NMR (CDCl3):
selected data; l 101.60 (C-1III), 101.33 (C-1II), 100.13
(C-1I), 92.47 (CCl3), 78.36 (C-3II), 77.36 (C-3III), 74.70
(C-5II), 73.69, 72.48, 72.34 (CH2Ph), 73.50 (C-3I), 73.21
(C-4I), 71.89 (C-4II), 69.66, 68.79 (C-2I, C-2III), 68.58
(C-6II), 67.12, 66.34 (C-5I, C-5III), 67.02 (C-4II), 56.23
(OCH3), 55.38 (C-2II), 52.76, 52.58 (COOCH3), 40.41
(CH2Cl), 38.81 ((CH3)3C), 26.88 ((CH3)3C); ISMS: m/z
1376, [M+NH4]+ for 35Cl. Anal. Calcd for
C65H69Cl4NO22: C, 57.48; H, 5.12; N, 1.04. Found: C,
57.35; H, 5.21; N, 1.08.
1
(677 mg, 77% from 20); [h]D +11° (c 1, CHCl3); H
NMR (250 MHz, CDCl3): carbohydrate ring protons
(see Table 3); l 8.0–7.20 (m, 15 H, Ph), 4.68, 4.48 (2
ABq, 4 H, CH2Ph), 3.83 (s, 3 H, COOCH3), 3.51 (s, 3
H, OCH3), 2.74 (d, 1 H, J 10.5 Hz, HO-4II), 1.15 (s, 9
H, (CH3)3C); 13C NMR (CDCl3): selected data; l
101.61 (C-1II), 100.39 (C-1I), 92.34 (CCl3), 77.45 (C-3I),
74.38 (C-3II), 73.68, 72.15 (CH2Ph), 73.26 (C-5I), 69.10
(C-4I), 68.74 (C-6I), 68.60 (C-4II), 68.43 (C-5II), 68.06
(C-2II), 57.25 (OCH3), 55.68 (C-2II), 52.52 (COOCH3),
39.13 ((CH3)3C), 27.14 ((CH3)3C); ISMS: m/z 915,
[M+NH4]+ for 35Cl. Anal. Calcd for C42H48Cl3NO14:
C, 56.22; H, 5.39; N, 1.57. Found: C, 56.07; H, 5.53; N,
1.60.
Methyl O-[methyl 2-O-benzoyl-3-O-benzyl-4-O-(4-
methoxybenzyl)-h-
benzyl-2-deoxy-4-O-pi6aloyl-2-trichloroacetamido-i-
galactopyranosyl)-(1“4)-(methyl 2-O-benzoyl-3-O-ben-
zyl-h- -idopyranosid)uronate (26).—Compound 25 (950
L
-idopyranosyluronate]-(1“3)-(6-O-
D
-
L
mg, 0.7 mmol) was O-dechloroacetylated as described
Methyl O-(methyl 2-O-benzoyl-3-O-benzyl-h-
pyranosyluronate)-(1“3)-4-O-acetyl-6-O-benzyl-2-
deoxy - 2 - trichloroacetamido - i - - galactopyranoside
L-ido-
for the preparation of 22. A solution of the resulting
,
alcohol and 4 A powdered molecular sieves (0.5 g) in
D
anhyd CH2Cl2 (7 mL) was stirred for 1 h at rt under
dry Ar. Solutions of Me3SiOTf (1 M in toluene, 115
mL) and 4-methoxybenzyl trichloroacetimidate (1 M in
toluene, 2 mL) were successively added dropwise, and
the mixture was stirred for 1 h at rt, then was neutral-
ized with Et3N (33 mL), filtered and concentrated. The
residue was eluted from a column (40 g) of silica gel
with 1:1 EtOAc–petroleum ether containing 0.2% of
Et3N to provide 26 as a white foam (667 mg, 68% from
25); [h]D −25° (c 1, CHCl3); 1H NMR (500 MHz,
CDCl3): carbohydrate ring protons (see Table 4); l
8.10–7.20 (m, 29 H, aromatic H), 4.67, 4.64, 4.38, 4.24
(4 ABq, 8 H, CH2Ph), 3.81 (s, 3 H, OCH3), 3.75, 3.71
(2 s, 6 H, COOCH3), 3.45 (s, 3 H, OCH3), 0.84 (s, 9 H,
(CH3)3C); ISMS: m/z 1419, [M+NH4]+, 1370, [M−
OCH3]+ for 35Cl. Anal. Calcd for C71H76Cl3NO22: C,
60.83; H, 5.29; N, 1.00. Found: C, 60.63; H, 5.39; N,
1.08.
(23).—Imidate 21 (1.90 g, 1.76 mmol) was treated as
described for the preparation of 22 to give 23 as a white
foam (1.11 g, 79% from 21); [h]D +12° (c 1, CHCl3);
1H NMR (CDCl3): carbohydrate ring protons (see
Table 3); l 8.0–7.20 (m, 15 H, Ph), 4.71, 4.48 (2 ABq,
4 H, CH2Ph), 3.82 (s, 3 H, COOCH3), 3.46 (s, 3 H,
OCH3), 2.75 (d, 1 H, J 11.3 Hz, HO-4II), 1.78 (s, 3 H,
Ac); 13C NMR (CDCl3): selected data; l 100.93 (C-1II),
100.20 (C-1I), 92.28 (CCl3), 75.60 (C-3I), 74.64 (C-3II),
73.70, 72.41 (CH2Ph), 72.96 (C-5I), 69.44 (C-4I), 68.69
(C-6I), 68.37 (C-5II), 68.11 (C-4II), 67.69 (C-2II), 57.42
(OCH3), 56.23 (C-2I), 52.54 (COOCH3), 20.39
(COCH3); ISMS: m/z 942, [M+NH4]+ for 35Cl. Anal.
Calcd for C39H42Cl3NO14: C, 54.78; H, 4.95; N, 1.64.
Found: C, 54.84; H, 4.98; N, 1.70.
Methyl
O-(methyl
2-O-benzoyl-3-O-benzyl-4-O-
chloroacetyl - h -
L
- idopyranosyluronate) - (1“3) - (6 - O-
benzyl-2-deoxy-4-O-pi6aloyl-2-trichloroacetamido-i-
galactopyranosyl)- (1“4)-(methyl 2-O-benzoyl-3-O-
benzyl-h- -idopyranosid)uronate (25).—A mixture of
imidate 20 (1.10 g, 1 mmol), methyl (methyl 2-O-ben-
zoyl-3-O-benzyl-a-
-idopyranosid)uronate 2410 (609
mg, 1.5 mmol) and 4 A powdered molecular sieves (1 g)
in anhyd CH2Cl2 (15 mL) was stirred for 1 h at rt under
dry Ar, then cooled to −15 °C. A solution of
Me3SiOTf in toluene (1 M, 0.15 mL) was added, and
the mixture was stirred for 30 min at −15 °C. Triethy-
lamine (65 mL) was added, and the mixture was filtered
and concentrated. The residue was eluted from a
column (60 g) of silica gel with 3:2 petroleum ether–
EtOAc containing 0.1% of Et3N to give 25 as a white
D
-
Methyl O-[methyl 2-O-benzoyl-3-O-benzyl-4-O-(4-
methoxybenzyl) - h -
acetamido-6-O-benzyl-2-deoxy-4-O-pi6aloyl-i-
actopyranosyl)-(1“4)-(methyl 2-O-benzoyl-3-O-benzyl-
h- -idopyranosid)uronate (27).—A mixture of 26 (0.79
L
- idopyranosyluronate] - (1“3) - (2-
L
D
-gal-
L
L
,
g, 0.6 mmol), Bu3SnH (0.91 mL, 3.4 mmol), and AIBN
(100 mg) in dry benzene (12 mL) was stirred for 30 min
at rt under a stream of dry Ar, then was heated for 2 h
at 80 °C, cooled and concentrated. The residue was
stirred with petroleum ether (3×20 mL), and the crys-
talline residue was filtered off and washed with
petroleum ether to give 27 (676 mg, 92%); mp 172–
173 °C (from EtOAc–petroleum ether); [h]D −19° (c 1,
CHCl3); 1H NMR (500 MHz, CDCl3): carbohydrate

686
N. Barroca, J.-C. Jacquinet / Carbohydrate Research 337 (2002) 673–689
ring protons (see Table 4); l 8.10–7.20 (m, 29 H,
aromatic H), 4.78, 4.72, 4.64, 4.12 (4 ABq, 8 H,
CH2Ph), 3.79 (s, 3 H, OCH3), 3.78, 3.73 (2 s, 6 H,
COOCH3), 3.45 (s, 3 H, OCH3), 1.82 (s, 3 H, NAc),
0.84 (s, 9 H, (CH3)3C); ISMS: m/z 1316, [M+NH4]+,
1299, [M+H]+. Anal. Calcd for C71H79NO22: C, 65.68;
H, 6.13; N, 1.08. Found: C, 65.38; H, 6.10; N, 1.22.
CH2Cl2–MeOH, then from a column (20 g) of silica gel
with 6:2:1 EtOAc–MeOH–water, and finally from a
column (1×20 cm) of Sephadex SP C25 [Na+] with
5:5:1 CH2Cl2–MeOH–water to afford 30 as a white
1
powder (144 mg, 63%); [h]D −38° (c 1, MeOH); H
NMR (500 MHz, CD3OD): carbohydrate ring protons
(see Table 4); l 7.20–6.80 (m, 19 H, aromatic H),
4.85–4.45 (m, 8 H, CH2Ph), 3.83, 3.56 (2 s, 6 H,
OCH3), 2.16 (s, 3 H, NAc), 1.23 (s, 9 H, (CH3)3C).
Anal. Calcd for C55H63NNa4O26S2: C, 50.42; H, 4.85;
N, 1.07. Found: C, 50.17; H, 5.05; N, 1.11.
[3-O-Benzyl-4-O-(4-methoxybenzyl)-h-
osyluronic acid]-(1“3)-(2-acetamido-6-O-benzyl-2-de-
oxy-4-O-pi6aloyl-i- -galactopyranosyl)-(1“4)-methyl
3-O-benzyl-h- -idopyranosiduronic acid (28).—A solu-
L-idopyran-
D
L
tion of 27 (658 mg, 0.5 mmol) in THF (15 mL) was
treated at 0 °C with 30% H2O2 (2.6 mL) and LiOH (1
M, 5.2 mL), and the mixture was stirred for 1 h at 0 °C
and for 15 h at rt, then was cooled to 0 °C. Methanol
(5 mL) and NaOH (4 M, 1.2 mL) were added, and the
mixture was stirred for 5 h at rt, then was diluted with
MeOH (15 mL) and treated with Amberlite IR–120
[H+] resin to pH 3.5 (pH meter control), filtered and
concentrated. The residue was eluted from a column
(40 g) of silica gel with 12:1“9:1“4:1 CH2Cl2–MeOH
to give 28 as a white foam (412 mg, 76%); [h]D −35° (c
1, CHCl3); 1H NMR (500 MHz, CD3OD): carbohy-
drate ring protons (see Table 4); l 7.40–6.90 (m, 19 H,
aromatic H), 4.63–4.45 (m, 8 H, CH2Ph), 3.80, 3.45 (2
s 6 H, OCH3), 2.08 (s, 3 H, NAc), 1.10 (s, 9 H,
(CH3)3C); ISMS: m/z 1080, [M+NH4]+. Anal. Calcd
for C55H67NO20: C, 62.19; H, 6.36; N, 1.32. Found: C,
61.91; H, 6.52; N, 1.18.
Sodium O-[sodium 3-O-benzyl-4-O-(4-methoxyben-
zyl)-2-O-sodium
(1“3)-(2-acetamido-6-O-benzyl-2-deoxy-4-O-sodium
sulfonato-i- -galactopyranosyl)-(1“4)-(methyl 3-O-
benzyl-2-O-sodium sulfonato-h- -idopyranosid)uronate
sulfonato-h-L-idopyranosyluronate]-
D
L
(31).—A mixture of 29 and sulfur trioxide–trimethy-
lamine complex (0.68 g, 4.9 mmol) in anhyd DMF (6
mL) was treated as described for the preparation of 30
to give 31 as a white foam (150 mg, 70%); [h]D −28° (c
1
1, MeOH); H NMR (500 MHz, CD3OD): carbohy-
drate ring protons (see Table 4); l 7.50–6.90 (m, 19 H,
aromatic H), 4.78–4.30 (m, 8 H, CH2Ph), 3.75, 3.37 (2
s, 6 H, OCH3), 2.07 (s, 3 H, NAc). Anal. Calcd for
C50H54NNa5O28S3: C, 45.22; H, 4.10; N, 1.05. Found:
C, 44.96; H, 4.25; N, 1.01.
Sodium O-[sodium 3-O-benzyl-4-O-(4-methoxyben-
zyl)-2-O-sodium
(1“3)-(2-acetamido-6-O-benzyl-2-deoxy-i-
pyranosyl)-(1“4)-( methyl 3-O-benzyl-2-O-sodium sul-
fonato-h- -idopyranosid)uronate (32).—A mixture of 30
sulfonato-h-
L
-idopyranosyluronate]-
D
-galacto-
[3-O-Benzyl-4-O-(4-methoxybenzyl)-h-
osyluronic acid]-(1“3)-(2-acetamido-6-O-benzyl-2-de-
oxy-i- -galactopyranosyl)-(1“4)-methyl 3-O-benzyl-
h- -idopyranosiduronic acid (29).—Compound 27 (389
L-idopyran-
L
D
(144 mg, 0.11 mmol) and NaOH (4 M, 5 mL) in MeOH
(8 mL) was stirred for 96 h at rt, then was treated with
Amberlite IR–120 [H+] resin to pH 7.0 (pH meter
control), filtered and concentrated. The residue was
eluted from a column (10 g) of silica gel with 6:2:1
EtOAc–MeOH–water to give 32 as a white foam (92
mg, 72%); [h]D −45° (c 1, MeOH); 1H NMR (500
MHz, CD3OD): carbohydrate ring protons (see Table
4); l 7.50–6.80 (m, 19 H, aromatic H), 4.85–4.30 (m, 8
H, CH2Ph), 3.76, 3.34 (2 s, 6 H, OCH3), 2.15 (s, 3 H,
NAc). Anal. Calcd for C50H55NNa4O25S2: C, 48.98; H,
4.52; N, 1.14. Found: C, 48.61; H, 4.75; N, 1.18.
L
mg, 0.3 mmol) was treated as described for the prepara-
tion of 28, but the mixture was stirred for 96 h at rt
with 4 M NaOH. After deionization, the residue was
eluted from a column (20 g) of silica gel with 10:2:1“
7:2:1 EtOAc–MeOH–water to give 29 as a white foam
1
(192 mg, 66%); [h]D −62° (c 1, MeOH); H NMR (500
MHz, CD3OD): carbohydrate ring protons (see Table
4); l 7.40–6.80 (m, 19 H, aromatic H), 4.75–4.41 (m, 8
H, CH2Ph), 3.75, 3.38 (2 s, 6 H, OCH3), 2.02 (s, 3 H,
NAc); ISMS: m/z 996, [M+NH4]+, 979, [M+H]+.
Anal. Calcd for C50H59NO19: C, 61.40; H, 6.08; N, 1.43.
Found: C, 61.12; H, 6.32; N, 1.24.
Sodium O-[sodium 2-O-sodium sulfonato-h-
ranosyluronate] - (1“3) - (2 - acetamido - 2 - deoxy - 4 - O-
sodium sulfonato-i- -galactopyranosyl)-(1“4)-(methyl
2-O-sodium sulfonato-h- -idopyranosid)uronate (1).—A
L-idopy-
Sodium O-[sodium 3-O-benzyl-4-O-(4-methoxyben-
D
zyl)-2-O-sodium
(1“3)-(2-acetamido-6-O-benzyl-2-deoxy-4-O-pi6al-
oyl-i- -galactopyranosyl)-(1“4)-( methyl 3-O-benzyl-
2-O-sodium sulfonato-h- -idopyranosid)uronate (30).—
sulfonato-h-
L
-idopyranosyluronate]-
L
solution of 31 (150 mg, 0.11 mmol) in 2:1 MeOH–wa-
ter (6 mL) was hydrogenated in the presence of 10%
Pd–C (100 mg) for 96 h at rt. The mixture was filtered
and concentrated, then was eluted from a column
(1.5×30 cm) of Sephadex LH–20 with water and
freeze-dried to afford amorphous hygroscopic 1 (98 mg,
D
L
A mixture of 28 (185 mg, 0.17 mmol) and sulfur
trioxide–trimethylamine complex (526 mg, 3.8 mmol)
in anhyd DMF (5 mL) was stirred for 72 h at 50 °C,
then was cooled. Methanol (4 mL) was added, and the
mixture was concentrated. The residue was eluted from
a column (2×50 cm) of Sephadex LH–20 with 1:1
1
93%); [h]D −21° (c 1, water); H NMR (500 MHz,
D2O, internal water, lH 4.754): carbohydrate ring pro-
tons (see Table 4); l 3.36 (s, 3 H, OCH3), 2.02 (s, 3 H,

N. Barroca, J.-C. Jacquinet / Carbohydrate Research 337 (2002) 673–689
687
NAc); 13C NMR (D2O, internal acetone, lC 30.45): l
176.46, 175.80, 175.15 (CꢀO), 103.13 (C-1II), 100.99
(C-1III), 100.32 (C-1I), 79.03 (C-4I), 76.97 (C-3II), 76.54
(C-4II), 76.15 (C-5II), 75.21, 74.30 (C-2I, C-2III), 73.24
(C-4I), 69.18, 69.08, 68.55, 67.59 (C-5I, C-5III, C-3I,
C-3III), 61.72 (C-6II), 55.92 (OCH3), 52.55 (C-2II), 23.52
(COCH3). Anal. Calcd for C21H28NNa5O27S3·H2O: C,
26.39; H, 3.16; N, 1.46. Found: C, 26.15; H, 3.27; N,
1.32.
(1“3)-4-O-acetyl-6-O-benzyl-2-deoxy-2-trichloroac-
etamido-i- -galactopyranoside (35).—A mixture of al-
D
cohol 23 (747 mg, 0.87 mmol) and imidate 33 (877 mg,
1.3 mmol) was treated as described for the preparation
of 24. The residue was eluted from a column (80 g) of
silica gel with 3:2 petroleum ether–EtOAc containing
0.2% of Et3N to give 35 as a white foam (910 mg, 75%);
1
[h]D +5° (c 1, CHCl3); H NMR (500 MHz, CDCl3):
carbohydrate ring protons (see Table 5); l 8.05–7.20
(m, 25 H, Ph), 4.66, 4.55, 4.48, 4.35 (4 ABq, 8 H,
CH2Ph), 3.73 (s, 3 H, COOCH3), 3.50 (s, 3 H, OCH3),
1.72, 1.65 (2 s, 6 H, Ac); ISMS: m/z 1402, [M+NH4]+
for 35Cl. Anal. Calcd for C63H66Cl6N2O20: C, 54.67; H,
4.81; N, 2.02. Found: C, 54.51; H, 4.87; N, 2.04.
Methyl O-(2-acetamido-4-O-acetyl-3,6-di-O-benzyl-
Sodium O-[sodium 2-O-sodium sulfonato-h-
ranosyluronate]-(1“3)-(2-acetamido-2-deoxy-i-
actopyranosyl)-(1“4)-(methyl 2-O-sodium sulfonato-h-
-idopyranosid)uronate (2).—Compound 32 (87 mg, 70
L
-idopy-
D
-gal-
L
mmol) was treated as described for the preparation of 1
to give amorphous hygroscopic 2 (57 mg, 96%); [h]D
1
−9° (c 1, water); H NMR (500 MHz, D2O, internal
2-deoxy-i-
D
-galactopyranosyl)-(1“4)-(methyl
2-O-
water, lH 4.754): carbohydrate ring protons (see Table
4); l 3.30 (s, 3 H, OCH3), 2.0 (s, 3 H, NAc); 13C NMR
(D2O, internal acetone, lC 30.45): l 176.69, 175.92,
175.25 (CꢀO), 103.64 (C-1II), 101.75 (C-1III), 100.41
(C-1I), 80.59 (C-4I), 79.01 (C-3II), 75.77 (C-5II), 74.36,
74.22 (C-2I, C-2III), 69.31, 69.22, 68.73, 68.57, 68.46,
67.65 (C-4II, C-4I, C-5I, C-5III, C-3I, C-3III), 61.88 (C-
6II), 55.96 (OCH3), 51.99 (C-2II), 23.43 (COCH3). Anal.
Calcd for C21H29NNa4O24S2·H2O: C, 29.55; H, 3.66; N,
1.64. Found: C, 29.27; H, 3.75; N, 1.51.
benzoyl-3-O-benzyl-h-
acetamido-6-O-benzyl-2-deoxy-4-O-pi6aloyl-i-D -gal-
L
-idopyranosyluronate)-(1“3)-2-
actopyranoside (36).—A mixture of 34 (885 mg, 0.62
mmol) and Bu3SnH (2 mL, 7.4 mmol) was treated as
described for the preparation of 27. The residue was
eluted from a column (25 g) of silica gel with 7:2
CH2Cl2–EtOAc to give 36 as a white foam (672 mg,
1
84%); [h]D +14° (c 1, CHCl3); H NMR (500 MHz,
CDCl3): carbohydrate ring protons (see Table 5); l
8.0–7.20 (m, 25 H, Ph), 4.62, 4.59, 4.47, 4.37 (4 ABq, 8
H, CH2Ph), 3.85 (s, 3 H, COOCH3), 3.49 (s, 3 H,
OCH3), 1.95, 1.92, 1.75 (3 s, 9 H, Ac), 1.14 (s, 9 H,
(CH3)3C); ISMS: m/z 1237, [M+NH4]+, 1188, [M−
OCH3]+. Anal. Calcd for C66H78N2O20: C, 65.00; H,
6.44; N, 2.30. Found: C, 64.81; H, 6.41; N, 2.29.
Methyl O-(2-acetamido-4-O-acetyl-3,6-di-O-benzyl-
Methyl O-(4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-tri-
chloroacetamido-i-
2 - O - benzoyl - 3 - O - benzyl - h -
(1“3)-6-O-benzyl-2-deoxy-4-O-pi6aloyl-2-trichloroac-
etamido-i- -galactopyranoside (34).—A mixture of al-
D
-galactopyranosyl)-(1“4)-(methyl
L
- idopyranosyluronate)-
D
cohol 22 (677 mg 0.75 mmol) and 4-O-acetyl-3,6-di-O-
benzyl-2-deoxy-2-trichloroacetamido-1-O-trichloroace-
2-deoxy-i-
D
-galactopyranosyl)-(1“4)-(methyl
2-O-
timidoyl-a-
D
-galactopyranose 3310 (756 mg, 1.12 mmol)
benzoyl-3-O-benzyl-h-
L
-idopyranosyluronate)-(1“3)-2-
was treated as described for the preparation of 24. The
residue was eluted from a column (60 g) of silica gel
with 2:1 petroleum ether–EtOAc containing 0.2% of
Et3N to give 34 as a white foam (855 mg, 80%); [h]D
+2° (c 1, CHCl3); 1H NMR (500 MHz, CDCl3): carbo-
hydrate ring protons (see Table 5); l 8.0–7.20 (m, 25
H, Ph), 4.66, 4.63, 4.47, 4.35 (4 ABq, 8 H, CH2Ph), 3.76
(s, 3 H, COOCH3), 3.52 (s, 3 H, OCH3), 1.82 (s, 3 H,
Ac), 1.15 (s, 9 H, (CH3)3C); 13C NMR (CDCl3): se-
lected data; l 101.74 (C-1II), 101.30 (C-1III), 100.34
(C-1I), 92.53, 92.30 (CCl3), 76.73 (C-3I), 75.17 (C-3II),
74.34 (C-3III), 73.69, 73.60, 72.25, 71.65 (CH2Ph), 73.26
(C-5I), 73.08 (C-4I) 71.90, 79.17, 68.66, 67.84, 67.19,
67.15, 65.12 (C-4I, C-4III, C-5III, C-6I, C-6III, C-2II,
C-5II), 57.32 (OCH3), 56.01, 55.44 (C-2I, C-2III), 52.64
(COOCH3), 39.11 ((CH3)3C), 27.17 ((CH3)3C), 20.44
(COCH3); ISMS: m/z 1444, [M+NH4]+ for 35Cl.
Anal. Calcd for C66H72Cl6N2O20: C, 55.59; H, 5.09; N,
1.97. Found: C, 55.48; H, 5.10; N, 1.92.
acetamido-4-O-acetyl-6-O-benzyl- 2-deoxy-i-D-gal-
actopyranoside (37).—Compound 35 was treated as
described for the preparation of 36. The residue was
eluted from a column (40 g) of silica gel with 10:1
CH2Cl2–MeOH to give 37 as a white foam (483 mg,
1
88%); [h]D +18° (c 1, CHCl3); H NMR (500 MHz,
CDCl3): carbohydrate ring protons (see Table 5); l
8.05–7.20 (m, 25 H, Ph), 4.70–4.35 (m, 8 H, CH2Ph),
3.74 (s, 3 H, COOCH3), 3.52 (s, 3 H, OCH3), 2.05, 1.98,
1.82, 1.71 (4 s, 12 H, Ac); ISMS: m/z 1200, [M+Na]+,
1146, [M−OCH3]+. Anal. Calcd for C63H72N2O20: C,
64.27; H, 6.16; N, 2.38. Found: C, 64.18; H, 6.14; N,
2.41.
Methyl O-(2-acetamido-3,6-di-O-benzyl-2-deoxy-i-
D
-galactopyranosyl)-(1“4)-(3-O-benzyl-h-
osyluronic acid)-(1“3)-2-acetamido-6-O-benzyl-2-de-
oxy-4-O-pi6aloyl-i- -galactopyranoside (38).—Com-
L-idopyran-
D
pound 36 (632 mg, 0.52 mmol) was treated as described
for the preparation of 28. The residue was eluted from
a column (40 g) of silica gel with 10:1“6:1 CH2Cl2–
MeOH to give 38 as a white foam (513 mg, 95%); [h]D
−5° (c 1, MeOH); 1H NMR (500 MHz, CD3OD):
Methyl O-(4-O-acetyl-3,6-di-O-benzyl-2-deoxy-2-tri-
chloroacetamido-i-
2 - O - benzoyl - 3 - O - benzyl - h -
D
-galactopyranosyl)-(1“4)-(methyl
- idopyranosyluronate)-
L

688
N. Barroca, J.-C. Jacquinet / Carbohydrate Research 337 (2002) 673–689
carbohydrate ring protons (see Table 5); l 7.60–7.20
(m, 20 H, Ph), 4.85–4.40 (m, 8 H, CH2Ph), 3.49 (s, 3 H,
OCH3), 2.05, 1.97 (2 s, 6 H, NAc), 1.14 (s, 9 H,
(CH3)3C); ISMS: m/z 1082, [M+Na]+, 1077, [M+
NH4]+, 1060, [M+H]+. Anal. Calcd for C56H70N2O18:
C, 63.50; H, 6.66; N, 2.64. Found: C, 63.32; H, 6.71; N,
2.51.
Methyl O-(2-acetamido-2-deoxy-4-O-sodium sulfo-
nato-i- -galactopyranosyl)-(1“4)-(sodium 2-O-sodium
sulfonato-h- -idopyranosyluronate)-(1“3)-2-acetami-
do-2-deoxy-4-O-sodium sulfonato-i- -galactopyran-
D
L
D
oside (3).—Compound 40 (340 mg, 0.26 mmol) was
treated as described for the preparation of 1 and freeze-
dried to give amorphous hygroscopic 3 (215 mg, 88%);
[h]D −20° (c 1, water); 1H NMR (500 MHz, D2O,
internal water, lH 4.754): carbohydrate ring protons
(see Table 5); l 3.42 (s, 3 H, OCH3), 2.15, 2.13 (2 s, 6
H, NAc); 13C NMR (D2O, internal acetone, lC 30.45):
l 175.81, 175.78, 174.85 (CꢀO), 103.95 (C-1III), 102.51
(C-1I), 101.18 (C-1II), 77.87 (C-3I), 77.45 (C-3III), 76.54
(C-4II), 76.41 (C-4III), 75.16 (C-4I), 75.05 (C-2II), 73.39
(C-5III), 70.95 (C-5I), 68.76 (C-5II), 67.78 (C-3II), 61.71,
61.57 (C-2I, C-2III), 57.80 (OCH3), 22.29, 22.05
(COCH3). Anal. Calcd for C23H34N2Na4O26S3·H2O: C,
28.75; H, 3.57; N, 2.91. Found: C, 28.47; H, 3.71; N,
2.52.
Methyl O-(2-acetamido-3,6-di-O-benzyl-2-deoxy-i-
D
-galactopyranosyl)-(1“4)-(3-O-benzyl-h-
osyluronic acid)-(1“3)-2-acetamido-6-O-benzyl-2-de-
oxy-i- -galactopyranoside (39).—(a) Compound 37
L-idopyran-
D
(470 mg, 0.4 mmol) was treated as described for the
preparation of 28. The residue was eluted from a
column (40 g) of silica gel with 10:1“6:1 CH2Cl2–
MeOH to give 39 as a white foam (373 mg, 95%); [h]D
1
−27° (c 1, MeOH); H NMR (500 MHz, CD3OD):
carbohydrate ring protons (see Table 5); l 7.50–7.10
(m, 20 H, Ph), 4.76–4.50 (m, 8 H, CH2Ph), 3.44 (s, 3 H,
OCH3), 2.12, 1.96 (2 s, 6 H, NAc); 13C NMR
(CD3OD): selected data; l 176.95, 176.55, 174.47
(CꢀO), 106.29 (C-1III), 104.90 (C-1I), 104.09 (C-1II),
81.98 (C-3I), 80.30 (C-3III), 79.47 (C-4II), 77.33 (C-3II),
75.66, 75.37 (C-5I, C-5III), 75.10, 73.84, 72.89, 71.26
(CH2Ph), 71.03, 70.27, 68.73, 66.48 (C-2II, C-5II, C-4I,
C-4III), 62.99, 62.97 (C-6I, C-6III), 57.71 (OCH3), 53.44,
53.40 (C-2I, C-2III), 24.69, 24.02 (COCH3); ISMS: m/z
993, [M+NH4]+, 944, [M−OCH3]+. Anal. Calcd for
C51H62N2O17: C, 62.81; H, 6.41; N, 2.87. Found: C,
62.66; H, 6.61; N, 2.77.
References
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(b) A mixture of 38 (467 mg, 0.45 mmol) and NaOH
(4 M, 10 mL) in MeOH (10 mL) was stirred for 96 h at
rt, then was worked-up as described for the preparation
of 28 to give 39 (268 mg, 61%); [h]D −27°(c 1, MeOH);
ISMS: m/z 993, [M+NH4]+.
Methyl O-(2-acetamido-3,6-di-O-benzyl-2-deoxy-4-
O-sodium
(sodium 3-O-benzyl-2-O-sodium sulfonato-h-
osyluronate)-(1“3)-2-acetamido-6-O-benzyl-2-deoxy-
4-O-sodium sulfonato-i- -galactopyranoside (40).—A
sulfonato-i-
D
-galactopyranosyl)-(1“4)-
L
-idopyran-
7. Jacquinet, J.-C.; Sinay¨, P. Carbohydr. Res. 1987, 159,
229–253.
D
8. Marra, A.; Dong, X.; Petitou, M.; Sinay¨, P. Carbohydr.
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mixture of 39 (389 mg, 0.4 mmol) and sulfur trioxide–
trimethylamine complex (2.20 g, 15.8 mmol) in anhyd
DMF (8 mL) was treated and purified as described for
the preparation of 30 to give 40 as a white powder (340
mg, 65%); [h]D +2° (c 1, MeOH); 1H NMR (500 MHz,
CD3OD): carbohydrate ring protons (see Table 5); l
7.65–7.30 (m, 20 H, Ph), 4.80–4.56 (m, 8 H, CH2Ph),
3.46 (s, 3 H, OCH3), 2.18, 2.04 (2 s, 6 H, NAc); 13C
NMR (CD3OD): selected data; l 175.97, 175.64, 175.02
(CꢀO), 105.76 (C-1III), 104.10 (C-1I), 102.35 (C-1II),
80.63 (C-3I), 79.56 (C-3III), 79.37 (C-4II), 77.88 (C-3II),
77.24, 75.48 (C-5I, C-5III), 75.09, 74.98, 74.02, 73.85
(CH2Ph), 73.59 (C-4I), 72.97 (C-2II), 72.58 (C-4III),
71.14 (C-5II), 63.05, 62.85 (C-6I, C-6III), 57.94 (OCH3),
53.40, 53.35 (C-2I, C-2III), 24.61, 24.26 (COCH3). Anal.
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2.15. Found: C, 46.70; H, 4.59; N, 2.07.
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