Chemical synthesis of sulfated oligosaccharides with a β-D-Gal-(1→3)-[β-D-Gal-(1→4)-(α-L-Fuc-(1→3)-β-D-GlcNAc-(1→6)]-α-D -GalNAc sequence

Article abstract of DOI:10.1016/S0008-6215(00)00211-1

The syntheses of two sulfated pentasaccharides: β-D-Gal6SO₃Na-(1→3)-[β-D-Gal-(1→4)-α-L-Fuc-(1→3)-β-D-GlcNAc-(1→6) ]-α-D-GalNAc→OMe (1) and β-D-Gal6SO₃Na-(1→3)-[β-D-Gal-(1→4)-α-L-Fuc-(1→3)-β-D-GlcNAc6SO₃Na -(1→6)]-α-D-GalNA

Full text of DOI:10.1016/S0008-6215(00)00211-1

www.elsevier.nl/locate/carres
Carbohydrate Research 329 (2000) 561–577
Chemical synthesis of sulfated oligosaccharides with a
b-
D
-Gal-(1“3)-[b-
D
-Gal-(1“4)-(a-
L
-Fuc-
(1“3)-b-
D
-GlcNAc-(1“6)]-a-
D
-GalNAc sequence
Jie Xia, T. Srikrishnan, James L. Alderfer, R.K. Jain, Conrad F. Piskorz,
Khushi L. Matta *
Molecular and Cellular Biophysics, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo,
NY 14263, USA
Received 22 May 2000; accepted 7 July 2000
Abstract
The syntheses of two sulfated pentasaccharides: b-
NAc-(1“6)]-a- -GalNAc“OMe (1) and b-
D
D
-Gal6SO3Na-(1“3)-[b-
D
D
-Gal-(1“4)-a-
L
L
-Fuc-(1“3)-b-
D
D
-Glc-
-Glc-
D
-Gal6SO3Na-(1“3)-[b-
-Gal-(1“4)-a-
-Fuc-(1“3)-b-
NAc6SO3Na-(1“6)]-a-
D
-GalNAc“OMe (2) by using LewisX trisaccharides 12 and 16 as glycosyl donors are
described. Sulfated oligosaccharides 1–2 and intermediate compounds are fully characterized by 2D 1H–1H
DQF-COSY and 2D ROESY experiments. © 2000 Elsevier Science Ltd. All rights reserved.
Keywords: Chemical synthesis; 2D DQF-COSY; 2D ROESY; Sulfated oligosaccharides; Crystallography
natural selectin ligands (GlyCAM-1 and
PSGL-1) carry sialyl LeX or NeuAca-(2“3)-
1. Introduction
Galb-(1“3)-GalNAc oligosaccharide unit as
Mammalian lectins mediate the early steps
part of core 2 O-linked structures, much effort
of recruitment of leukocytes from the blood-
has been made for the synthesis of sialylated
and sulfated LewisX, and mimics of SLeX [5,6].
Recently, Kiessling and co-workers [7] have
stream in a variety of normal and pathological
situations [1–4]. In the last decade there has
been a tremendous interest in the synthesis of
high affinity molecules that target the selectin
family of cell-surface proteins (L, P and E)
because these molecules can become future
drugs for treatment of various diseases such as
chronic and acute inflammation. Since the
examined the inhibition of -selectin with var-
L
ious disulfated LeX derivatives. We became
interested in the synthesis of sialyl and sul-
fated core 2 analogs for the binding studies of
these selectins because the role of core 2
analogs as inhibitors for these selectins has
become evident [8]. Our interest in core 2
branched structures which contain sulfates has
also stemmed from our study of sulfotrans-
ferase using the core 2 branched structures as
acceptors [9]. We hereby report the synthesis
of branched structures 1 and 2.
* Corresponding author. Tel.: +1-716-8452300; fax: +1-
716-8453458.
E-mail address: klmatta@yahoo.com (K.L. Matta).
0008-6215/00/$ - see front matter © 2000 Elsevier Science Ltd. All rights reserved.
PII: S0008-6215(00)00211-1

562
J. Xia et al. / Carbohydrate Research 329 (2000) 561–577
2. Results and discussion
glycosylated with donor 6 in the presence of
silver triflate–tin chloride as catalyst [10] to
give disaccharide 7 in good yield (73%). The
stereochemical outcome of both the glycosylic
and position connectivity of disaccharide 7
was well established through homonuclear
correlation experiments (2D 1H–1H DQF-
COSY: Fig. 1 and 2D ROESY: Fig. 2). The
(1“4)-linkage of disaccharide 7 was indicated
by the observation of a strong NOE cross
peak between H-1II and H-4I in the 2D
ROESY spectrum. A 1,2-trans glycoside of
disaccharide 7 was confirmed by a larger cou-
Synthesis of trisaccharide building blocks 12
and 16.—In order to synthesize two sulfated
oligosaccharides 1 and 2,
key building blocks 12, 16, 21 and 27 were
prepared starting from monosaccharide build-
ing blocks
3
pling constant of J1,2 (J 8.0 Hz). The struc-
ture of disaccharide
7
was further
characterized by X-ray diffraction analysis
(Fig. 3).
Disaccharide 7 was fucosylated with methyl
2,3,4-tri-O-benzyl-1-thio-b- -fucoside (3) in
L
the presence of tetrabutylammonium bro-
mide–copper bromide as catalyst [11], afford-
ing trisaccharide 8 in good yield (76%). The
removal of benzyl groups from compound 8
was achieved in the presence of 10% palla-
dium-on-carbon catalyst under a hydrogen at-
mosphere. Compound 9 was then treated with
1:1 acetic anhydride–pyridine in the presence
according to Schemes 1–3.
The HO-4 of diol 5 was regioselectively
,
Scheme 1. Reagents and conditions. (a) 5:1 AgOTf–SnCl2/CH2Cl2–toluene, 4 A 10% MS, −15 to 0 °C, 12 h, 73%; (b) 3,
,
CuBr2–n-Bu4NBr, 5:1 ClCH2CH2Cl–DMF, 4 A MS, rt, 16–24 h, 91%; (c) 10% H2/Pd–C, 1:1 CH2Cl2–CH3OH, rt, 12 h, 96%;
(d) 1:1 Ac2O–pyridine, DMAP, rt, 12 h, 96%; (e) H2SO4–Ac2O, rt, 12 h, 76%; (f) PhSH/CH2Cl2, BF3·Et2O, rt, N2, 16 h, 67%.
,
Scheme 2. Reagents and conditions: (a) AgOTf–SnCl2/5:1 CH2Cl2–toluene, 4 A MS, −15 to 0 °C, 12 h, 75%; (b) 3,
,
CuBr2–n-Bu4NBr, 5:1 ClCH2CH2Cl–DMF, 4 A MS, rt, 16–24 h, 76%.

J. Xia et al. / Carbohydrate Research 329 (2000) 561–577
563
1
Fig. 1. 400 MHz 2D H–1H DQF-COSY spectrum of disaccharide 7 recorded in CDCl3 at 303.0 K.
Fig. 2. 400 MHz 2D ROESY spectrum of disaccharide 7 recorded in CDCl3 at 303.0 K.

564
J. Xia et al. / Carbohydrate Research 329 (2000) 561–577
Scheme 3. Reagents and conditions: (a) BnBr/KOH, 18-crown-6, THF, rt, 12 h, 86%; (b) 1 N HCl–1,4-dioxane, 4 h, 75%; (c) 1:1
,
pyridine–Ac2O, 65–70 °C, 3 h, 76%; (d) Me3SiBr/BiBr3, CH2Cl2, rt, 12 h, 59%; (e) Hg(CN)2–HgBr2/ClCH2CH2Cl, 4 A MS, rt,
12 h, 66%; (f) 60% HOAc, 60–65 °C, 1.5 h, 76%.
of a catalytic amount of DMAP, providing
compound 10. The conversion of compound
10 into compound 11 was successfully per-
formed by treatment of 10 with acetic anhy-
periments. For preparation of trisaccharide
donor 16, a strategy of sequential glycosyla-
tion is employed. Use of three different leav-
ing groups (fluorine, SMe and SPh) which
exhibit different reactivities in different cou-
pling reactions allows us to carry out the
direct glycosylation of monosaccharide accep-
tor 13. The construction of trisaccharide
donor 16 was outlined in Scheme 2. Regiose-
lective glycosylation of diol 13 with donor 14
was performed in the presence of silver tri-
flate–tin chloride as catalyst to afford disac-
dride [12] and
a
catalytic amount of
concentrated sulfuric acid. Trisaccharide
donor 12 was obtained by treatment of com-
pound 11 with thiophenol in the presence of
BF3Et2O [13] under a nitrogen atmosphere
(Scheme 1). The structure of trisaccharide
donor 12 was fully confirmed by a combina-
tion of 2D DQF-COSY and 2D ROESY ex-
Fig. 3. An ORTEP [22] diagram showing the molecular structure of disaccharide 7.

J. Xia et al. / Carbohydrate Research 329 (2000) 561–577
565
The structure of trisaccharide 16 was estab-
lished by 2D NMR experiments (2D DQF-
COSY and 2D ROESY). The a- -fucopy-
L
ranoside of trisaccharide 16 was indicated by a
3
small coupling constant of J1,2 (J 3.3 Hz),
which is a characteristic feature of a 1,2-cis-a-
L
-fucopyranoside.
Synthesis of disaccharide building block
21.—Disaccharide building block 27 was pre-
pared according to documented protocol [14].
The construction of disaccharide acceptor 21
is illustrated in Scheme 3. Compound 17 was
prepared according to literature procedures
[15]. Monosaccharide 17 was treated with
potassium hydroxide in the presence of a
phase-transfer catalyst [16] (18-crown-6) in
dry THF, followed by benzyl bromide, to
afford compound 18 in excellent yield (86%).
Compound 18 was treated with 1 N HCl in
1,4-dioxane at reflux, resulting in removal of
the O-isopropylidene group from 18. The re-
Scheme 4. Reagents and conditions: (a) NIS–TfOH (cat)/
CH2Cl2, 4 A MS, −65 to −60 °C, 3–4 h, 44%; (b) 1:1
,
Ac2O–pyridine, DMAP, rt, 12 h, 86%; (c) 1:5
NH2NH2·xH2O–CH3OH, 85–90 °C, 4 h, then, 1:1 Ac2O–
pyridine, DMAP, rt, 12 h, 76%; (d) H2, 10% Pd–C, CH2Cl2–
CH3OH, rt, 6–8 h, quantitative; (e) SO3·pyridine–pyridine,
0–25 °C, 6 h, 79%; (f) 1 M, 1:1 CH3ONa–CH3OH (cat)/
CH3OH–water, rt, 24 h, 36%.
sulting 6-O-benzyl- -galactose was further
D
acetylated with 1:1 acetic anhydride–pyridine
at 65–70 °C to give 6-O-benzyl-1,2,3,4-tetra-
O-acetyl- -galactopyranose, which was then
D
treated with bromotrimethylsilane in the pres-
ence of bismuth(III) bromide [17] to give
donor 19. Disaccharide 21a was obtained by
treatment of compound 20 with glycosyl
donor 19 in the presence of mercury cyanide
and mercury bromide as catalysts [18]. Com-
pound 21a was treated with 60% acetic acid at
60–65 °C to give the desired disaccharide ac-
ceptor 21 in good yield (76%).
The construction of target sulfated oligosac-
charides 1 and 2.—The target sulfated
oligosaccharides 1 and 2 were constructed as
shown in Schemes 4 and 5. On account of the
much higher reactivity of the primary hy-
droxyl group in acceptors 21 and 27, totally
regioselective glycosylation of the primary
HO-6 of disaccharide acceptor 21 with trisac-
charide donor 12 was achieved under con-
trolled glycosylation conditions to give 22 as
the only glycosylation product. The b-(1“6)-
linkage of 22 was indicated by observation of
strong NOE cross peaks between H-1III of
sugar residue III and H-6aI, H-6bI of sugar
residue I by 2D ROESY spectroscopy. Com-
pound 22 was acetylated with 1:1 acetic anhy-
dride–pyridine in the presence of a catalytic
Scheme 5. Reagents and conditions: (a) NIS–TfOH (cat)/
CH2Cl2,
,
4 A MS, −75 to −70 °C, 1.5 h, 46%;
(b)HN2CSNH2/2,6-lutidine, 60–65 °C, 18 h, 87%; (c)
SO3·pyridine–pyridine, 0–5 °C, 12 h, 79%; (d) DDQ/4:1
CH2Cl2–CH3OH, rt, 20 h, 79%; (e) SO3·pyridine–pyridine,
0–5 °C, 12 h, 78%; (f) H2/10% Pd–C, 4:1 CH2Cl2–MeOH, rt,
10 h, 87%; (g) 1:5 NH2NH2·xH2O–CH3OH, 85–90 °C, 4–5
h, then, 1:1 Ac2O–pyridine, DMAP, rt, 12 h, 76%; (h) 1 M,
CH3ONa–CH3OH (cat)/1:1 CH3OH–water (1:1), rt, 12 h,
26%.
charide 15 in good yield (75%). Disaccharide
15 was further fucosylated with donor 3 to
give trisaccharide 16 in excellent yield (76%).

566
J. Xia et al. / Carbohydrate Research 329 (2000) 561–577
Compound 28 was obtained in reasonable
yield (46%). The b-(1“6)-linkage of com-
pound 28 was indicated by observation of
strong NOE cross peaks between H-1III of
sugar residue III and H-6aI, H-6bI of sugar
residue I by 2D ROESY spectroscopy. The
a-bromoacetyl group was removed by treat-
ment of compound 28 with thiourea and 2,6-
lutidine at reflux in dichloromethane to give
compound 29. Compound 29 was then treated
with the sulfur trioxide–pyridine complex in
dry pyridine to give compound 30. The next
reaction was the selective removal of the 2-
naphthymethyl (NAP) protecting group from
compound 30. The NAP protecting group was
recently introduced as a hydroxyl protecting
group [19] and can be removed by DDQ in
the presence of benzyl groups [20]. Thus, com-
pound 30 was carefully treated with DDQ,
resulting in the removal of NAP protecting
group. Compound 31 was then treated with
the sulfur trioxide–pyridine complex in dry
pyridine to give 32. Compound 32 was depro-
tected by the following procedure to give
target oligosaccharide 2: (a) removal of benzyl
amount of DMAP to give compound 23.
Compound 23 was then treated with 1:5
NH2NH2·H2O–methanol at 85–90 °C, which
resulted in the removal of the phthalimido
group, followed with 1:1 acetic anhydride–
pyridine in the presence of a catalytic amount
of DMAP at room temperature to afford
compound 24. Compound 24 was treated with
10% palladium-on-carbon in a hydrogen at-
mosphere to give compound 25. Compound
26 was obtained by treatment of compound 25
with sulfur trioxide–pyridine complex in dry
pyridine. O-Deacetylation of 26 was per-
formed by treatment with 1 M sodium
methoxide to give compound 1. The structure
of compound 1 was well characterized by 2D
1H–1H DQF-COSY, 2D ROESY (Fig. 4), and
13C NMR spectroscopy and by FABMS. Sim-
ilarly, the synthesis of target oligosaccharide 2
was carried out as outlined in Scheme 5. Re-
gioselective glycosylation of HO-6 of diol 27
was achieved by treatment of diol 27 with
trisaccharide donor 16 under controlled glyco-
sylation conditions.
Fig. 4. 600 MHz 2D ROESY spectrum of pentasaccharide 1 recorded in D2O at 303.0 K.

J. Xia et al. / Carbohydrate Research 329 (2000) 561–577
567
protecting groups; (b) removal of the phthal-
imido group, followed with acetic anhydride–
pyridine and DMAP; (c) removal of acetyl
and trimethylacetyl groups, which is shown in
Scheme 5. Target oligosaccharide 2 was well
For ROESY experiments, the mixing time was
set at 400 ms. All samples submitted for ele-
mental analyses were dried under vacuum
over P2O5 at rt. Elemental analyses were car-
ried out by Robertson Laboratory, Madison,
NJ. p-Toluenesulfonic acid monohydrate (p-
TsOH·H2O) was dried by co-evaporating with
dry MeCN three times at 80 °C before use.
Dichloromethane, MeCN, MeOH, benzene,
1
characterized with 2D H–1H DQF-COSY,
13
2D ROESY, and C NMR spectroscopy and
by FABMS.
In summary, we describe the convergent
syntheses of two sulfated pentasaccharides 1
and 2 through a potentially general strategy of
regio- and stereoselective glycosylation by the
use of unprotected or partially protected
acceptors.
,
and DMF were kept dry over 4 A MS, pyri-
dine was redistilled over KOH, and ni-
tromethane was freshly distilled over P2O5.
Methyl (2,3,4,6-tetra-O-acetyl-i-
pyranosyl) - (1“4) - 6 - O - benzyl - 2 - deoxy - 2-
phthalimido-i- -glucopyranoside (7).—A soln
D
-galacto-
D
of compound 5 (1.18 g, 3.38 mmol), com-
pound 6 (2.37 g, 5.75 mmol), and SnCl2 (1.13
g, 4.39 mmol) in dry 5:1 CH2Cl2–toluene (43
3. Experimental
,
mL) containing 4 A MS (8 g) was stirred at
General procedures.—TLC was conducted
on glass plates precoated with a 0.25-mm layer
of Silica Gel 60 F-254 (Analtech, GHLF uni-
plates). The components were located either
by exposure to UV light or by spraying with
10% H2SO4, and 0.2% p-anisaldehyde in
EtOH soln. Solutions were concd under re-
duced pressure. The silica gel used for column
chromatography was Baker Analyzed (60–200
mesh ASTM). Optical rotations were mea-
sured at 25 °C with a Perkin–Elmer 241 po-
−15 °C for 2 h under a N2 atmosphere. Silver
triflate (832 mg, 4.39 mmol) was added, and
the mixture was stirred overnight at −15 to
0 °C under a N2 atmosphere, then neutralized
with Et3N. The solids were filtered off, and the
organic layer was washed with satd NaHCO3
soln, water, and dried (Na2SO4). The organic
solvent was evaporated under reduced pres-
sure to give a crude mixture, which was then
applied to a column of silica gel and eluted
with 1:1 hexane–EtOAc to give pure com-
pound 7 (1.84 g, 73%) as a white solid. Rf 0.34
(1:1 hexane–EtOAc); [h]D +8.8° (c 0.5,
CHCl3); 1H NMR (CDCl3, 400 MHz): l
7.84–7.82 (m, 2 H, ArH), 7.72–7.70 (m, 2 H,
ArH), 7.41–7.32 (m, 5 H, ArH), 5.33–5.32 (d,
1 H, J 3.2 Hz, H-4II), 5.21–5.16 (dd, 1 H, J
1
larimeter. H NMR spectra were recorded at
303.0 K with either a Bruker AM 400 (400
MHz) or AMX 600 (600 MHz) spectrometer.
The values of l (ppm) are given relative to the
signal (l 0) for internal Me4Si for solns in
CDCl3, CD2Cl2, or CD3OD. 13C NMR spectra
were recorded at 303.0 K with a Bruker AM
400 (100.6 MHz) spectrometer using CDCl3
(77.0 ppm), CD2Cl2 (54.15 ppm), CD3OD
(49.15 ppm), or acetone-d6 (206.0 or 29.8
ppm) as reference. First-order chemical shifts
and coupling constants (J/Hz) were obtained
from one-dimensional spectra, and assign-
ments of protons were based on 2D DQF
1H–1H COSY and 2D ROESY experiments.
Two-dimensional double-quantum filtered
phase sensitive 1H–1H correlated spectra
8.0, 10.4 Hz, H-2II), 5.13–5.11 (d, 1 H, J
1, 2
8.8 Hz, H-1I), 4.95–4.92 (dd, 1 H, J 3.6, 10.2
Hz, H-3II), 4.77–4.74 (d, 1 H, Jgem 12.4 Hz,
OCHAPh, ABq), 4.55–5.52 (d, 1 H, Jgem 12.0
Hz, OCHBPh, ABq), 4.50–4.48 (d, 1 H, J
1,2
8.0 Hz, H-1II), 4.42–4.37 (t, 1 H, H-3I), 4.18–
4.13 (dd, 1 H, J 8.8 Hz, H-2I), 4.06–4.04 (m,
2 H, H-6bII, H-6aII), 3.95 (d, 1 H, OH-3I),
3.91–3.88 (t, 1 H, J 7.2, 6.4 Hz, H-5II), 3.77–
3.69 (m, 3 H, H-5I, H-4I, H-6bI), 3.65–
3.62 (m, 1 H, H-6aI), 3.44 (s, 3 H, OCH3), 2.11
(s, 3 H, Ac), 2.00 (s, 3 H, Ac), 1.95, (s, 3 H,
Ac), 1.91 (s, 3 H, Ac); 13C NMR (CDCl3,
100.6 MHz): l 170.46 (CꢀO), 170.06 (CꢀO),
169.92 (CꢀO), 169.15 (CꢀO), 138.18, 133.99,
1
(DQF H–1H COSY), rotating-frame nuclear
Overhauser
enhancement
spectroscopy
(ROESY) were recorded at 303.0 K using a
Bruker AM 400 (400 MHz) spectrometer and
a Bruker AMX 600 (600 MHz) spectrometer.

568
J. Xia et al. / Carbohydrate Research 329 (2000) 561–577
Table 1
(Ac), 20.56 (Ac), 20.50 (Ac), 20.33 (Ac); Anal
Calcd for C36H41NO16: C, 58.12; H, 5.56; N,
1.88. Found: C, 58.00; H, 5.59; N, 1.81.
X-ray crystallography.—Crystals of 7 were
obtained by a slow evaporation of the com-
pound dissolved in hexanes–EtOAc. The crys-
tals were rectangular needles. A crystal of
dimensions 0.15×0.45×0.25 mm was used
for the X-ray diffraction. The crystals be-
longed to a monoclinic system, space group
P21. Table 1 gives the crystal data and struc-
ture refinement parameters.
X-ray crystal data and structure refinement for 7
Compound
Methyl (2,3,4,6-tetra-
-galactopy-
ranosyl)-(1“4)-6-O-
benzyl-2-deoxy-2-phthalimido-
b-D-glucopyranoside (7)
white/rectangular needle
C36H41NO16
743.7
2293
monoclinic
P21
O-acetyl-b-
D
Color/shape
Empirical formula
Formula weight
Temperature (°C)
Crystal system
Space group
Unit cell dimensions
Complete three-dimensional data were col-
lected on an Enraf–Nonius CAD-4 computer-
controlled diffractometer. A total of 4099
reflections (2qmax=150°) were collected by the
ꢀ/2q method, out of which 3820 were consid-
ered significant (I]3|). The crystal structure
was solved by a routine and straightforward
application of the SHELX-96 program on a
Silicon Graphics R10,000 computer. The
structure was refined using the SHELX-96
package [21] of programs. All hydrogen atoms
were located on a difference Fourier map.
Refinements were carried out with anisotropic
thermal parameters for the nonhydrogen
atoms and isotropic thermal parameters for
the hydrogen atoms. The final reliability index
(R-factor) was 0.0879 for all the observed
3892 reflections and 0.064 for 3186 reflections
(I]4|) with a goodness of fit parameter S=
1.120. Table 2 gives the final positional
parameters for the nonhydrogen atoms in the
structure. The crystal structural investigation
confirms the structure of compound 7. The
disaccharide has the sugar residues, the galac-
topyranosyl and the glucopyranoside in the
4C1 chair conformation. The ring-to-ring con-
formation can be described as (ƒ, „)=(159.7,
−126.3). The conformation across the C-5ꢁC-
6 bond is trans–gauche for the galactopyra-
nosyl ring and gauche–gauche for the
glucopyranosyl ring. The crystal structure is
stabilized by a network of OꢁH···O hydrogen
bonds.
,
a (A)
12.529 (3)
14.765 (5)
10.728 (3)
90.0
113.98 (2)
90.0
1813.3(9)
2 (two molecules per unit
,
b (A)
,
c (A)
h (°)
i (°)
k (°)
3
,
Volume (A )
Z
cell)
1.362
0.915
Dcalcd (g cm−3
Absorption coefficient
(mm−1
Difftractometer
Radiation (u=1.5418 A)
F(000)
Crystal size (mm)
2q Range for data
collection (°)
)
)
CAD-4
Cu Ka
784
0.15×0.45×0.25
0–150
,
Index ranges (°)
−105h510, 05k517,
−185l518
4099
Reflections collected
Independent/observed
reflections
Absorption correction
Range of relative
transmission factors
Refinement method
3892 (3820 with I53|)
semiempirical from psi scans
0.99 and 0.90
full-matrix least-squares on
F2
SHELXS-96
Computing
Data/restraints/parameters
Goodness-of-fit on F2
Function minimized
Final R indices
3820/0/643
1.120
S[ꢀF2oꢀ−(1/k)ꢀF2cꢀ]
0.0879 for all 3892
reflections
0.064 for 3186 [I]4|(I)]
Final extinction coefficient 5.59×10−7
Large difference peak and
hole (e A
0.15 and −0.15
−3
,
)
Methyl (2,3,4,6-tetra-O-acetyl-i-
pyranosyl)-(1“4)-[(2,3,4-tri-O-benzyl-h-
fucopyranosyl)-(1“3)]-6-O-benzyl-2-deoxy-2-
phthalimido-i- -glucopyranoside (8).—A soln
of compound 7 (320 mg, 0.36 mmol), methyl
2,3,4-tri-O-benzyl-1-thio-b- -fucopyranoside
D
-galacto-
L
-
131.92, 128.55, 127.91, 127.86, 123.34, 101.54,
99.26, 82.06, 74.27, 73.74, 71.22, 70.81, 69.69,
68.80, 67.97, 66.90, 61.47, 56.75, 55.94, 20.72
D
L

J. Xia et al. / Carbohydrate Research 329 (2000) 561–577
569
Table 2
(3, 338 mg, 0.72 mmol), and n-Bu4NBr (232
Final fractional positional parameters for nonhydrogen atoms
with estimated standard deviations given in parentheses for 7
mg, 0.72 mmol) in dry 5:1 1,2-
,
dichloroethane–DMF (15 mL) containing 4 A
a
MS (2 g) was stirred for 2 h at rt under a N2
atmosphere. Copper bromide (161 mg, 0.72
mmol) was added, and the mixture was stirred
overnight at rt. An additional portion of com-
pound 3 (790 mg), CuBr2 (101 mg) and n-
Bu4NBr (145 mg) were added, and the stirring
was continued for a total of 24 h. The solids
were filtered off, the organic layer was washed
with satd NaHCO3 soln, and water, and then
it was dried (Na2SO4) and concd. The crude
product was applied to a short column of
silica gel and eluted with 2:1 hexane–EtOAc
to give compound 8 (380 mg, 0.32 mmol,
91%) as a white solid. Rf 0.17 (2:1 hexane–
Atom
x
y
z
Ueq
O-1
O-2
O-3
O-4
O-5
O-6
O-7
O-8
0.0784(3) 0.5082(3)
0.1769(3) 0.3972(2)
−0.0120(3) 0.7273(3)
0.0887(4) 0.8505(3)
−0.0927(4) 0.3807(3)
−0.0191(4) 0.2016(3)
0.2190(4) 0.2272(3)
0.3041(5) 0.7413(5)
0.2848(4) 0.5702(3)
−0.1547(4) 0.5666(4)
−0.0860(5) 0.1083(3)
0.1886(5) 0.0781(3)
0.3417(4) 0.8014(4)
−0.0515(7) 0.3525(7)
0.3475(8) 0.3787(6)
0.7758(4)
0.9223(4)
0.8543(4)
0.9695(4)
0.6207(4)
0.6622(4)
0.8293(5)
1.2883(5)
1.0130(5)
0.8231(5)
0.7786(7)
0.8058(7)
0.8940(5)
0.4401(6)
1.1611(9)
0.0480(8)
0.0487(9)
0.0508(9)
0.0523(9)
0.0587(10)
0.0553(10)
0.0595(11)
0.0765(15)
0.0598(10)
0.0711(13)
0.0686(13)
0.0840(20)
0.0714(13)
0.1130(20)
0.1220(30)
O-9
O-10
O-11
O-12
O-13
O-14
O-15
O-16
N-1
C-1
C-2
C-3
C-4
1
0.2713(11) 0.3584(11) 1.3027(11) 0.1760(50)
EtOAc); [h]D +5.4° (c 0.7, CHCl3); H NMR
0.2987(4) 0.7553(3)
0.0170(5) 0.2641(4)
0.0793(5) 0.5834(3)
0.1733(6) 0.3142(4)
0.1874(5) 0.7107(4)
0.0977(5) 0.7733(4)
0.0652(5) 0.4242(4)
0.1981(5) 0.6268(4)
0.1329(5) 0.2379(4)
−0.0132(5) 0.6506(4)
0.0228(5) 0.3563(4)
0.3474(6) 0.7676(4)
−0.0712(8) 0.1259(6)
0.0335(8) 0.9240(4)
0.5617(6) 0.8844(5)
−0.3636(6) 0.5634(4)
0.3661(5) 0.7984(4)
0.2945(7) 0.3002(5)
0.4672(5) 0.8372(4)
0.2375(6) 0.1436(5)
0.4576(5) 0.8172(4)
0.6384(7) 0.8890(7)
1.0723(5)
0.7725(6)
0.8617(6)
0.9879(6)
1.0002(6)
0.9023(6)
0.8260(6)
0.9215(6)
0.8859(6)
0.7745(6)
0.7127(6)
1.2134(6)
0.0466(10)
0.0499(12)
0.0441(11)
0.0520(13)
0.0442(11)
0.0458(11)
0.0475(12)
0.0486(12)
0.0507(12)
0.0503(12)
0.0491(12)
0.0547(14)
(CDCl3, 600 MHz): l 7.67–7.66 (m, 2 H,
ArH), 7.41–7.39 (m, 2 H, ArH), 7.26–7.01
(m, 20 H, ArH), 5.23–5.22 (d, J1,2 3.2 Hz,
H-4II), 5.01–4.99 (m, 2 H, H-2II, H-1I, J1,2 8.4
Hz), 4.85–4.73 (m, 4 H, OCHPh, H-1III,
OCHPh, H-3II), 4.71–4.62 (m, 2 H, H-1II,
H-3I), 4.61–4.59 (dd, 2 H, OCH2Ph), 4.58–
4.55 (d, 1 H, Jgem 12.0 Hz, OCHPh, ABq),
4.51–4.48 (d, 1 H, Jgem 12.0 Hz, OCHPh,
ABq), 4.42–4.33 (m, 2 H, OCHPh, H-2I),
4.26–4.23 (d, 1 H, Jgem 12.0 Hz, ABq), 4.15–
4.09 (m, 2 H, H-4I, H-6bII), 4.00–3.94 (dd, 1
H, H-6aII), 3.92–3.80 (m, 4 H, H-6aI, H-3III,
H-6bI, H-2III), 3.61 (d, 1 H, J 3.2 Hz, H-4III),
3.55–3.45 (m, 2 H, H-5I, H-5II), 3.28 (s, 3 H,
OCH3), 2.01 (s, 3 H, Ac), 2.00 (s, 3 H, Ac),
1.94 (s, 3 H, Ac), 1.84 (s, 3 H, Ac), 1.21–1.19
(d, 3 H, J 6.4 Hz, CHI3II); 13C NMR (CDCl3,
100.6 MHz): l 170.56 (CꢀO), 170.36 (CꢀO),
170.09 (CꢀO), 169.84 (CꢀO), 168.79 (CꢀO),
139.93, 139.85, 138.89, 138.62, 134.93, 134.82,
129.28, 128.78, 128.66, 128.59, 128.51, 128.07,
127.83, 127.79, 124.19, 100.47, 99.95, 98.32,
80.42, 78.90, 75.97, 75.81, 75.34, 75.26, 74.18,
73.56, 73.16, 72.94, 71.69, 71.30, 69.58, 68.53,
67.63, 67.18, 61.21, 57.25, 56.97, 21.90 (Ac),
21.87 (Ac), 21.67 (Ac), 20.83 (Ac), 16.89
(CH3); Anal Calcd for C63H68NO20: C, 65.27;
H, 5.91; N, 1.21. Found: C, 64.57; H, 5.94; N,
1.11.
C-5
C-6
C-7
C-8
C-9
C-10
C-11
C-12
C-13
C-14
C-15
C-16
C-17
C-18
C-19
C-20
C-21
C-22
C-23
C-24
C-25
C-26
C-27
C-28
C-29
C-30
C-31
C-32
C-33
C-34
C-35
C-36
0.6711(11) 0.0640(20)
0.8793(8)
1.1259(9)
0.7549(8)
1.0152(6)
1.0978(9)
1.1276(7)
0.7931(8)
1.2462(7)
0.0620(20)
0.0680(20)
0.0610(20)
0.0516(13)
0.0720(20)
0.0553(14)
0.0670(20)
0.0561(14)
1.3696(10) 0.0830(20)
−0.2430(14) 0.3994(12) 0.4055(12) 0.1130(50)
−0.1078(10) 0.0681(7)
−0.1212(8) 0.3753(6)
−0.5262(8) 0.6569(7)
0.5539(10) 0.0870(30)
0.4867(7) 0.0770(20)
0.6200(10) 0.0850(20)
0.3151(11) 0.3873(11) 1.2405(14) 0.1330(60)
−0.5626(8) 0.6626(7)
−0.4034(8) 0.5695(6)
−0.4275(7) 0.6083(6)
−0.2567(6) 0.5123(6)
0.6463(7) 0.9103(6)
0.5421(6) 0.8417(6)
0.7244(11) 0.0880(20)
0.8603(9)
0.6365(8)
0.0760(20)
0.0740(20)
0.7767(10) 0.0750(20)
1.2502(10) 0.0760(20)
1.3724(8)
0.0740(20)
0.3241(14) 0.1464(10) 0.7335(20) 0.1250(70)
0.3829(11) 0.4979(7)
−0.5004(9) 0.6193(7)
−0.1354(6) 0.6116(5)
1.3227(10) 0.0890(20)
0.8430(11) 0.0890(20)
0.7199(7)
0.0604(15)
a Ueq is defined as the trace of the orthogonalized Uij
tensor.
Phenyl (2,3,4,6-tetra-O-acetyl-i-
pyranosyl)-(1“4)-[(2,3,4-tri-O-acetyl-h-
D
-galacto-
L
-

570
J. Xia et al. / Carbohydrate Research 329 (2000) 561–577
170.20 (CꢀO), 169.90 (CꢀO), 169.15 (CꢀO),
134.75, 133.0, 132, 129.00, 128.25, 124.00,
101.00, 95.50, 84.30, 77.30, 75.50, 72.55, 71.89,
71.45, 71.40, 69.00, 68.30, 67.88, 66.89, 64.50,
62.20, 61.00, 56.50, 21.60 (Ac), 21.20 (Ac),
20.98 (Ac), 20.88 (Ac), 20.84 (Ac), 20.81(Ac),
20.70 (2 Ac), 16.45 (CH3); Anal Calcd for
C48H55NO23S: C, 55.10; H, 5.30; N, 1.34, S,
3.07. Found: C, 55.38; H, 5.29; N, 1.26, S,
2.94.
fucopyranosyl)-(1“3)]-6-O-acetyl-2-deoxy-2-
phthalimido-1-thio-i- -glucopyranoside (12).
D
—A soln of compound 8 (310 mg, 0.32
mmol), and 10% Pd–C (320 mg) in a mixture
of MeOH (5 mL) and CH2Cl2 (5 mL) was
stirred overnight at rt under a H2 atmosphere.
The catalyst was filtered off, and the filtrate
was concd under reduced pressure to give
crude 9, which was then treated with 1:1
Ac2O–pyridine (6 mL) in the presence of a
catalytic amount of DMAP (3 mg) at rt
overnight. The reaction mixture was then
concd. The crude product was applied to a
column of silica gel and eluted with 1:2 hex-
ane–EtOAc to give pure compound 10 (250
mg, 0.21 mmol, 96%) in two steps. To a cold
(0 °C) soln of compound 10 (320 mg, 0.33
mmol) in Ac2O (3.6 mL), was added Ac2O
(3.6 mL) and concd H2SO4 (80 mL). The reac-
tion mixture was stirred for 16–20 h at the
same temperature, then neutralized with satd
NaHCO3 soln and concd to give a crude
residue, which was extracted with CH2Cl2 (30
mL). The organic extract was washed with
satd NaCl soln (3×100 mL), dried over an-
hyd Na2SO4 and concd to give a crude
product. BF3Et2O (120 mL) was added to a
soln of the crude product and thiophenol (0.3
mL) in dry CH2Cl2 (6 mL), and the mixture
was stirred for 16 h at rt under a N2 atmo-
sphere, then neutralized with Et3N and concd
to a mixture, which was applied to a column
of silica gel and eluted with 1:1 hexane–
EtOAc to give pure compound 12 (260 mg,
67%) as an amorphous solid. [h]D −38.8° (c
Phenyl
(2,3,4-tri-O-acetyl-6-O-trimethyl-
-galactopyranosyl)-(1“4)-2-deoxy-
acetyl-i-
D
6-O-naphthymethyl-2-phthalimido-1-thio-i- -
D
glucopyranoside (15).—A soln of compound
13 (4.74 g, 8.73 mmol), compound 14 (6.83 g,
17.5 mmol), and SnCl2 (4.97 g, 26.2 mmol) in
dry 5:1 CH2Cl2–toluene (100 mL) containing
,
4 A MS (25 g) was stirred at −15 °C for
1.5–2 h under a N2 atmosphere. Silver triflate
(6.73 g, 26.2 mmol) was added, and the reac-
tion mixture was stirred overnight at −15 to
0 °C under a N2 atmosphere, then neutralized
with Et3N. The solids were filtered off, the
organic layer was washed with satd NaHCO3
soln, water, and then dried (Na2SO4). The
solvent was evaporated under reduced pres-
sure to give a crude mixture that was applied
to a column of silica gel and eluted with 1:1
hexane–EtOAc to give pure 15 (5.97 g, 75%)
as an amorphous solid. Rf 0.54 (1:1 hexane–
EtOAc); [h]D +35.0° (c 0.5, CHCl3); 1H
NMR (CDCl3, 400 MHz): l 7.86–7.80 (m, 6
H, ArH), 7.68–7.64 (m, 2 H, ArH), 7.52–7.40
(m, 5 H, ArH), 7.28–7.16 (m, 3 H, ArH),
5.65–5.63 (d, 1 H, J1,2 10.4 Hz, H-1I), 5.32–
5.31 (d, 1 H, J 2.0 Hz, H-4II), 5.20–5.16 (t, 1
H, J 10.0, 8.8 Hz, H-2II), 4.94–4.90 (dd, 1 H,
H-3II), 4.89–4.86 (d, 1 H, J 12.0 Hz,
OCHAC10H7, ABq), 4.72–4.69 (d, 1 H, J 12.0
Hz, OCHBC10H7, ABq), 4.53–4.51 (d, 1 H, J
8.0 Hz), 4.47–4.42 (t, 1 H), 4.05–4.03 (m, 4
H), 3.87–3.71 (m, 4 H), 2.14–2.10 (s, 3 H,
Ac), 1.96 (s, 3 H, Ac), 1.94 (s, 3 H, 2 Ac), 1.12
(s, 9 H, t-Bu); 13C NMR (CDCl3, 100.6 MHz):
l 177.90 (CꢀO), 169.93 (CꢀO), 169.83 (CꢀO),
169.20 (CꢀO), 168.30 (CꢀO), 16.84 (CꢀO),
135.63, 134.14, 133.09, 132.64, 128.91, 128.39,
127.94, 127.81, 126.71, 126.34, 126.09, 125.95,
123.60, 123.28, 100.42, 83.57, 81.51, 78.30,
73.82, 71.43, 70.79, 70.78, 68.80, 67.99, 66.69,
61.82, 55.17, 38.67, 27.09 (3 CH3), 20.63 (Ac),
1
0.5, CHCl3); H NMR (CDCl3, 600 MHz): l
7.75–7.00 (m 9 H, ArH), 5.55–5.40 (m, 2 H,
H-1I, J1,2 7.8 Hz, H-4II, J 2.8 Hz), 5.40–5.30
(d, 1 H, J 3.2 Hz, H-4III), 5.29–5.20 (m, 2 H,
H-3III, H-2II), 5.19–5.15 (dd, 1 H, H-3II),
5.15–5.10 (dd, 1 H, H-5III), 4.95–4.90 (d, 1 H,
J 3.1 Hz, H-1III), 4.85–4.80 (dd, 1 H, H-2III),
4.75–4.65 (m, 2 H, H-3I, H-6bI), 4.60–4.50
(m, 2 H, H-1II, H-6bII), 4.40–4.30 (m, 2 H,
H-2I, H-6bI), 4.20–4.19 (dd, 1 H, H-6aI),
4.00–3.90 (m, 2 H, H-4I, H-5II), 3.80–3.70
(dd, 1 H, H-5I), 2.30 (s, 3 H, Ac), 2.29 (s, 3 H,
Ac), 2.27 (s, 3 H, Ac), 1.99 (s, 3 H, Ac), 1.20
(d, 3 H, J 6.4 Hz, CHI3II); 13C NMR (CDCl3,
100.6 MHz): l 170.90 (CꢀO), 170.86 (CꢀO),
170.80 (CꢀO), 170.69 (CꢀO), 170.35 (CꢀO),

J. Xia et al. / Carbohydrate Research 329 (2000) 561–577
571
73.19, 72.52, 71.20, 70.60, 69.20, 68.05, 66.90,
66.74, 60.30, 55.76, 20.80 (3 Ac), 20.67 (Ac),
16.89 (CH3); Anal Calcd for C75H79NO19S·
H2O: C, 66.80; H, 5.90; N, 1.04, S, 2.38.
Found: C, 66.33; H, 6.02; N, 0.70, S, 2.24.
20.52 (Ac), 20.33 (Ac); Anal Calcd for
C48H51NO15S: C, 63.08; H, 5.62; N, 1.53.
Found: C, 62.90; H, 6.06; N, 1.98.
Phenyl
(2,3,4-tri-O-acetyl-6-O-trimethyl-
-galactopyranosyl)-(1“4)-[(2,3,4-
acetyl-i-
D
tri-O-benzyl-h- -fucopyranosyl)-(1“3)]-2-
L
2,3,4-Tri-O-acetyl-6-O-benzyl-h- -galacto-
D
deoxy-6-O-naphthymethyl-2-phthalimido-1-
thio-i- -glucopyranoside (16).—A soln of
compound 15 (5.2 g, 5.7 mmol), methyl 2,3,4-
-fucopyranoside (3, 10.63 g,
pyranosyl bromide (19).—Compound 18 was
obtained in quantitative yield by treatment of
compound 17 with BnBr in the presence of
powdered KOH and 18-crown-6 at rt for 4–5
h in dry THF. The removal of the O-iso-
propylidene group was achieved by treatment
of compound 18 with 1 N HCl in refluxing
1,4-dioxane, and the product was acetylated
with 1:1 pyridine–Ac2O and followed by
treatment of the acetylated compound with
BiBr3 as described in the literature [17]. After
purification by silica gel chromatography (2:1
hexane–EtOAc), compound 19 was obtained
D
tri-O-benzyl-b-
L
22.8 mmol), and n-Bu4NBr (7.36 g, 22.8
mmol) in dry 5:1 1,2-dichloroethane–DMF
,
(30 mL) containing 4 A MS (25 g) was stirred
for 2 h at rt under a N2 atmosphere. Copper
bromide (1.25 g, 5.6 mmol) was then added,
and the mixture was stirred for 16–24 h at the
same temperature. The solids were filtered off,
the organic layer washed with satd NaHCO3
soln, and water, dried (Na2SO4) and concd.
The crude product was applied to a column of
silica gel and eluted with 2:1 hexane–EtOAc
to give compound 16 (87%) as a white solid.
Rf 0.17 (2:1 hexane–EtOAc); [h]D +10.2° (c
1
in 59% yield. Rf 0.45 (2:1 hexane–EtOAc); H
NMR (CDCl3, 400 MHz): l 7.42–7.26 (m, 5
H, ArH), 6.76–6.72 (d, 1 H, H-1), 5.58–5.52
(d, 1 H, J 2.8 Hz, H-4), 5.44–5.36 (dd, 1 H,
H-2), 5.10–5.00 (dd, 1 H, H-3), 4.56–4.50 (d,
1 H, J 12.6 Hz, OCHPh, ABq), 4.50–4.40 (d,
1 H, OCHPh), 3.50–3.40 (m, 3 H, H-6a,
H-6b), 2.05 (s, 3 H, Ac), 2.03 (s, 3 H, Ac),
1.99 (s, 3 H, Ac); 13C NMR (CDCl3, 100.6
MHz): l 170.30 (CꢀO), 170.10 (CꢀO), 169.59
(CꢀO), 137.44, 128.73, 128.21, 108.51, 73.86,
72.65, 70.46, 69.31, 67.29, 67.05, 20.90 (Ac),
20.81 (Ac), 20.77 (Ac).
1
0.54, CHCl3); H NMR (CDCl3, 600 MHz): l
7.93–7.87 (m, 4 H, ArH), 7.70–7.60 (m, 3 H,
ArH), 7.56–7.44 (m, 4 H, ArH), 7.40–7.36
(m, 3 H, ArH), 7.28–7.16 (m, 14 H, ArH),
6.98–6.92 (m, 3 H, ArH), 5.50–5.45 (d, 1 H, J
7.8 Hz, H-1I), 5.24–5.18 (d, 1 H, J 2.8 Hz,
H-4III), 5.10–5.00 (dd, 1 H, H-2II), 4.97–4.92
(d, 1 H, Jgem 11.6 Hz, OCHAr, ABq), 4.88–
4.86 (dd, 1 H, J 3.6, 10.4 Hz, H-3II), 4.82–4.80
(m, 2 H, OCHAr, H-1III, J1,2 3.3 Hz), 4.80–
4.78 (d, 1 H, J1,2 7.6 Hz, H-1II), 4.74–4.70 (m,
2 H, H-3I, OCHAr, ABq), 4.68–4.60 (m, 2 H,
H-5III, Jgem 11.8 Hz, OCHAr, ABq), 4.40–4.32
(t, 1 H, J 10.4 Hz, H-2I), 4.32–4.20 (dd, 4 H,
2 OCH2Ar), 4.19–4.16 (t, 1 H, J 9.2 Hz,
H-4I), 4.14–4.00 (dd, 1 H, H-6bII), 4.00–3.84
(m, 4 H, H-6aII, H-6bI, H-6aI, H-3III), 3.80
(dd, 1 H, H-2III), 3.76–3.60 (m, 3 H, H-4III,
H-5I, H-5II), 1.94 (s, 3 H, Ac), 1.92 (s, 6 H, 2
Ac), 1.20 (d, 3 H, J 6.4 Hz, CHI3II), 1.16 (s, 9
H, t-Bu); 13C NMR (CDCl3, 100.6 MHz): l
170.09 (CꢀO), 169.94 (CꢀO), 168.89 (CꢀO),
138.84, 138.35, 135.49, 134.42, 133.43, 133.25,
132.65, 131.89, 129.04, 128.70, 128.38, 128.34,
128.27, 128.19, 128.15, 128.09, 128.03, 127.99,
127.61, 127.42, 127.34, 127.17, 126.80, 126.49,
126.21, 126.01, 123.84, 99.77, 97.85, 84.53,
79.95, 79.77, 75.29, 74.75, 74.40, 73.88, 73.83,
Methyl 2 - acetamido - 2 - deoxy - 4,6 - O - (p-
methoxybenzylidene) - h -
D
- galactopyranoside
(20).—p-Toluenesulfonic acid monohydrate
(769 mg) was added to a soln of methyl
2-acetamido-2-deoxy-a- -galactopyranoside
D
(3.35 g, 14.3 mol) and a,a-dimethoxy(p-
methyloxy)toluene (3.90 g, 21.39 mmol) in dry
MeCN (105 mL). The mixture was stirred
overnight at rt and quenched with Et3N, and
concd under reduced pressure to give a crude
product, which was then applied to a column
of silica gel and eluted with 30:1 CH2Cl2–
MeOH to give pure compound 20 (4.40 g,
87%) as a white solid. Rf 0.39 (1:1 hexane–
EtOAc); 1H NMR (CDCl3, 400 MHz): l
7.46–7.43 (d, 2 H, J 8.4 Hz, ArH), 6.91–6.89
(d, 2 H, J 8.0 Hz, ArH), 5.74–5.72 (d, 1 H, J
8.0 Hz, NHAc), 5.53 (s, 1 H, PhCHO2), 4.84–

572
J. Xia et al. / Carbohydrate Research 329 (2000) 561–577
5 H, ArH), 5.60–5.52 (m, 1 H, J 8.6 Hz,
NHAc), 5.44–5.40 (d, 1 H, J 3.0 Hz), 5.20–
5.12 (dd, 1 H), 5.04–4.96 (dd, 1 H), 4.80–4.72
(d, 1 H, J 3.2 Hz, H-1I), 4.60–4.56 (d, 1 H, J
7.8 Hz, H-1II), 4.54–4.48 (d, 1 H, Jgem 12.6
Hz, OCHPh, ABq), 4.48–4.40 (d, 1 H, Jgem
12.4 Hz, OCHPh, ABq), 4.10 (s, 1 H), 4.00–
3.80 (m, 3 H), 3.80–3.70 (m, 4 H), 3.60–3.45
(m, 3 H), 3.44 (s, 3 H, OCH3), 2.08 (s, 3 H,
Ac), 2.06 (s, 3 H, Ac), 1.99 (s, 3 H, Ac), 1.98
(s, 3 H, Ac); 13C NMR (CDCl3, 100.6 MHz):
l 170.50 (CꢀO), 170.30 (CꢀO), 169.70 (CꢀO),
128.75, 128.25, 128.00, 101.75, 99.30, 74.10,
72.85, 71.00, 69.83 (2 C), 69.21, 68.20, 67.85,
63.00, 55.21, 48.00, 23.60 (NAc), 20.90 (Ac),
20.83 (Ac), 20.75 (Ac); Anal Calcd for
C28H39NO14: C, 54.81; H, 6.41; N, 2.28.
Found: C, 54.28; H, 6.19; N, 1.98.
4.83 (d, 1 H, J 2.8 Hz, H-1), 4.48–4.43 (m, 1
H), 4.28–4.19 (m, 3 H), 4.07–4.04 (d, 1 H),
3.81 (s, 3 H, OCH3), 3.64 (s, 1 H), 3.40 (s, 3
H, OCH3), 1.98 (s, 3 H, Ac);); 13C NMR
(CDCl3, 100.6 MHz): l 160.91 (CꢀO), 130.34,
127.93, 113.87, 101.49, 99.67, 76.92, 69.54,
69.35, 63.05, 55.69, 55.56, 50.66, 23.67 (Ac).
Methyl (2,3,4-tri-O-acetyl-6-O-benzyl-i-
D
- galactopyanosyl) - (1“3) - 2 - acetamido - 2-
deoxy-h-
D
-galactopyranoside (21).—To a mix-
ture of compound 20 (2.20 g, 6.23 mmol),
powered Hg(CN)2 (7.6 g) and HgBr2 (760 mg)
,
in dry CH2Cl2 (60 mL) containing 4 A MS (20
g) was added 2,3,4-tri-O-acetyl-6-O-benzyl-a-
D
-galactopyranosyl bromide 19 (4.27 g, 9.30
mmol), and the mixture was stirred overnight
at rt. The reaction mixture was diluted with
benzene and washed with satd aq NaHCO3,
10% KI soln, and water. The organic layer
was dried with Na2SO4 and concd. The crude
residue was applied to a column of silica gel
and eluted with 20:1 CH2Cl2 –MeOH to give
compound 21a (2.3 g, 51%) as an amorphous
Methyl [(2,3,4,6-tetra-O-acetyl-i-
topyranosyl)-(1“4)-[(2,3,4-tri-O-acetyl-h-
fucopyranosyl)-(1“3)]-6-O-acetyl-2-deoxy-2-
phthalimido - i - - glucopyranosyl) - (1“6)-
[(2,3,4-tri-O-acetyl-6-O-benzyl-i- -galacto-
pyranosyl)]-(1“3)-2-acetamido-4-O-acetyl-2-
deoxy-h- -galactopyranoside (23).—A soln of
D
-galac-
L
-
D
D
1
solid. H NMR (CDCl3, 400 MHz): l 7.48–
7.42 (d, 2 H, J 8.7 Hz, ArH), 7.40–7.28 (d, 5
H, ArH), 6.96–6.88 (d, 2 H, J 8.6 Hz, ArH),
5.70–5.60 (d, 1 H, J 7.8 Hz, NHAc), 5.47 (s,
1 H, PhCHO2), 5.42–5.40 (d, 1 H, J 4.4 Hz,
H-4II), 5.20–5.10 (dd, 1 H, H-2II), 5.05–4.95
(dd, 1 H, H-3II), 4.80–4.70 (d, 1 H, J 4.0 Hz,
D
trisaccharide donor 12 (264 mg, 0.21 mmol),
disaccharide acceptor 21 (233 mg, 0.19 mmol),
and N-iodosuccinimide (NIS, 126 mg, 0.62
,
mmol) in dry CH2Cl2 (12 mL) containing 4 A
MS (8 g) was stirred for 2 h at −65 to
−60 °C under a N2 atmosphere. Trifluoro-
methanesulfonic acid (TfOH) (20 mL) in dry
CH2Cl2 (2 mL) was then added dropwise, and
the mixture was stirred for 2 h at the same
temperature. The mixture was then neutral-
ized with satd aq NaHCO3 soln. The solids
were filtered off, the organic layer was washed
with satd NaHCO3 soln, 10% Na2S2O3, and
water, and then it was dried (Na2SO4) and
concd. The crude mixture was applied to a
column of silica gel and eluted with 30:1
CH2Cl2–MeOH to give pure compound 22
(200 mg, 41%) as a glassy white solid. Com-
pound 22 was further acetylated with 1:1 pyri-
dine–Ac2O (10 mL) in the presence of a
catalytic amount of DMAP (5 mg). The soln
was concd and the crude product was purified
by silica gel column chromatography (30:1
CH2Cl2–MeOH) to give compound 23
in quantitative yield as an amorphous solid.
Rf 0.46 (30:1 CH2Cl2–MeOH); 1H NMR
H-1I), 4.70–4.60 (d, 1 H, J
8.0 Hz, H-1II),
1,2
4.56–4.44 (dd, 2 H, OCH2Ph, AB), 4.30–4.20
(d, 1 H), 4.20–4.10 (dd, 1 H), 4.00–3.85 (m, 3
H), 3.85–3.75 (m, 4 H, OCH3), 3.65–3.55 (dd,
1 H), 3.55–3.50 (m, 2 H), 3.42 (s, 3 H, OCH3),
2.08 (s, 3 H, Ac), 2.02 (s, 3 H, Ac), 1.95 (s, 3
H, Ac), 1.94 (s, 3 H, Ac); 13C NMR (CDCl3,
100.6 MHz): l 171.12 (CꢀO), 171.10 (CꢀO),
170.40 (CꢀO), 161.50 (CꢀO), 131.80, 129.50,
128.83, 128.80, 128.40, 114.50, 103.18, 101.72,
101.51, 76.71, 76.46, 74.44, 73.27, 72.22, 70.06,
69.82, 69.02, 68.60, 63.85, 56.22, 48.99, 24.17
(NAc), 21.39 (Ac), 21.33 (Ac), 21.13 (Ac);
Anal Calcd for C36H45NO15: C, 59.07; H, 6.20;
N, 1.92. Found: C, 58.97; H, 6.19; N, 1.61.
Compound 21a was treated with 60%
HOAc at 60–65 °C for 1.5 h and purified by
silica gel column to give pure compound 21
(76%) as an amorphous solid. Rf 0.47 (20:1
CH2Cl2–MeOH); [h]D +5.7° (c, 0.1, CHCl3);
1H NMR (CDCl3, 400 MHz): l 7.60–7.20 (m,

J. Xia et al. / Carbohydrate Research 329 (2000) 561–577
573
lated mixture was concd and passed through a
short column of silica gel and eluted with 30:1
CH2Cl2–MeOH to give compound 24 (65 mg,
86%) as an amorphous solid. [h]D −16.6° (c
(CDCl3, 600 MHz): l 7.88–7.68 (m, 4 H,
ArH), 7.40–7.24 (m, 5 H, ArH), 5.53–5.50 (d,
1 H, J 9.2 Hz, NHAc), 5.47–5.46 (d, 1 H, J
3.2 Hz, H-4II), 5.38–5.37 (d, 1 H, J 2.8 Hz,
H-4IV), 5.34–5.33 (d, 1 H, J 2.0 Hz, H-4V),
5.17–5.08 (m, 3 H, H-4I, H-3V, H-2II), 5.05–
4.88 (m, 6 H, H-2IV, H-1III, H-3II, H-5V, H-1V,
H-3IV), 4.81–4.75 (dd, 1 H, J 4.0, 11.0 Hz,
H-2V), 4.72–4.60 (m, 2 H, H-3III, H-6bIII),
4.57–4.40 (m, 5 H, OCHPh, ABq, H-1II, H-
1IV, H-6bII, OCHPh, ABq), 4.32–4.16 (m, 5
H, H-1I, H-6aII, H-2I, H-2III, H-6aIII), 3.97–
3.86 (m, 2 H, H-4III, H-5II), 3.84–4.71 (m, 4
H, H-6bI, H-3I, H-5I, H-5IV), 3.63–3.60 (dd, 1
H, H-5III), 3.56–3.48 (dd, 1 H, H-6bIV), 3.46–
3.40 (dd, 1 H, H-6aIV), 3.28–3.18 (dd, 1 H,
H-6aI), 2.89 (s, 3 H, OCH3), 2.18 (s, 3 H, Ac),
2.17 (s, 9 H, 3 Ac), 2.10 (s, 3 H, Ac), 2.09 (s,
3 H, Ac), 2.05 (s, 6 H, 2 Ac), 2.03 (s, 3 H, Ac),
2.02 (s, 3 H, Ac), 1.97 (s, 3 H, Ac), 1.90 (s, 3
H, Ac), 1.89 (s, 3 H, Ac), 1.21 (d, 3 H, J 6.4
Hz, CHV3 ); 13C NMR (CDCl3, 100.6 MHz): l
170.82 (CꢀO), 170.73 (CꢀO), 170.68 (CꢀO),
170.59 (CꢀO), 170.37 (CꢀO), 170.33 (CꢀO),
170.30 (CꢀO), 170.23 (CꢀO), 170.11 (CꢀO),
169.84 (CꢀO), 169.68 (CꢀO), 169.64 (CꢀO),
169.07 (CꢀO), 134.47, 128.66, 128.09, 127.99,
100.84, 100.80, 98.77, 98.29, 95.28, 75.36,
73.81, 73.76, 73.36, 72.85, 72.52, 71.54, 71.24,
71.06, 70.17, 69.92, 69.09, 68.51, 68.23, 67.99,
67.75, 67.50, 66.81, 64.41, 61.72, 60.87, 56.33,
54.70, 48.96, 31.02 (Ac), 23.46 (Ac), 20.96
(Ac), 20.92 (Ac), 20.87 (Ac), 20.80 (Ac), 20.76
(Ac), 20.64 (Ac), 15.99 (CH3); Anal Calcd for
C72H90N2O38: C, 54.34; H, 5.70; N, 1.76.
Found: C, 54.49; H, 5.81; N, 1.64.
1
0.2, CHCl3); H NMR (CDCl3, 600 MHz): l
7.40–7.25 (m, 5 H, ArH), 6.00–5.90 (d, 1 H, J
8.3 Hz, NHAc), 5.80–5.78 (d, 1 H, J 8.4 Hz,
NHAc), 5.43–5.37 (m, 4 H, H-1III, H-1V, H-
4II, H-4IV), 5.26–5.24 (d, 1 H, J 2.8 Hz, H-4I),
5.22–5.18 (dd, 1 H, J 3.2, 11.2 Hz), 5.12–4.95
(m, 5 H, H-2IV, H-2II, H-2V, H-3IV, H-3II),
4.95–4.88 (dd, 1 H, H-5V), 4.74–4.73 (d, 1 H,
J1,2 3.6 Hz, H-1I), 4.59–4.42 (m, 7 H, H-1II,
H-6bIV, OCHAPh, H-6bIII, H-1IV, H-2I,
OCHBPh), 4.32–4.29 (dd, 1 H, H-6aIII), 4.14–
4.10 (dd, 1 H, H-6aI), 4.00–3.99 (dd, 1 H,
H-3I), 3.97–3.93 (m, 1 H, H-5I), 3.87 (t, 1 H,
H-5III), 3.82–3.79 (m, 4 H, H-5II, H-6bI),
3.60–3.50 (m, 2 H, H-6bII, H-5IV), 3.50–3.40
(m, 2 H, H-6aII, H-6aI), 3.39 (s, 3 H, OCH3),
2.19 (s, 3 H, Ac), 2.18 (s, 3 H, Ac), 2.15 (s, 3
H, Ac), 2.09 (s, 3 H, Ac), 2.08 (s, 3 H, Ac),
2.07 (s, 3 H, Ac), 2.06 (s, 3 H, Ac), 1.98 (s, 3
H, Ac), 1.97 (s, 3 H, Ac), 1.95 (s, 3 H, Ac),
1.94 (s, 3 H, Ac), 1.69 (s, 6 H, 2 Ac), 1.24–
1.20 (d, 3 H, J 6.4 Hz, CHV3 ); 13C NMR
(CDCl3, 100.6 MHz): l 171.85 (CꢀO), 171.20
(CꢀO), 171.15 (CꢀO), 170.90 (CꢀO), 170.80
(CꢀO), 170.60 (CꢀO), 170.49 (CꢀO), 170.43
(CꢀO), 170.40 (CꢀO), 170.35 (CꢀO), 170.20
(CꢀO), 170.15 (CꢀO), 170.10 (CꢀO), 169.40
(CꢀO), 137.52, 128.83, 128.33, 128.25, 100.75
(2 C), 100.50, 98.90, 95.50, 74.85, 73.89, 73.25,
72.60, 72.20, 71.60, 71.40, 71.30, 71.20, 69.90,
69.25, 69.00, 68.60, 68.45, 68.25, 67.90, 67.60,
66.98, 64.50, 62.25, 61.00, 55.80, 49.85, 23.57
(NAc), 23.40 (NAc), 21.16 (Ac), 21.06 (Ac),
21.02 (2 Ac), 20.98 (Ac), 20.94 (Ac), 20.86 (2
Ac), 20.82 (2 Ac), 20.76 (Ac), 20.72 (Ac),
16.20 (CH3); Anal Calcd for C66H90N2O37: C,
52.73; H, 6.03; N, 1.86. Found: C, 53.16; H,
5.30; N, 1.56.
Methyl (2,3,4,6-tetra-O-acetyl-i-
pyranosyl)-(1“4)-[(2,3,4-tri-O-acetyl-h-
fucopyranosyl) - (1“3)] - 2 - acetamido - 6 - O-
acetyl-2-deoxy-i- -glucopyranosyl)-(1“6)-
[(2,3,4-tri-O-acetyl-6-O-benzyl-i- -galacto-
pyranosyl)-(1“3)]-2-acetamido-4-O-acetyl-2-
deoxy-h- -galactopyranoside (24).—Com-
D
-galacto-
L
-
D
D
D
Methyl (2,3,4,6-tetra-O-acetyl-i-
pyranosyl)-(1“4)-[(2,3,4-tri-O-acetyl-h-
fucopyranosyl) - (1“3)] - 2 - acetamido - 6 - O-
acetyl-2-deoxy-i- -glucopyranosyl)-(1“6)-
[(2,3,4-tri-O-acetyl-6-O-sulfo-i- -galactopyra-
nosyl)-(1“3)]-2-acetamido-4-O-acetyl-2-de-
oxy-h- -galactopyranoside sodium salt (26).—
D
-galacto-
pound 23 (80 mg, 0.05 mmol) in a mixture of
MeOH (5 mL) and NH2NH2·H2O (1 mL) was
stirred for 4–5 h at 80–85 °C. The reaction
mixture was concd under reduced pressure to
give a crude mixture, which was co-evapo-
rated with toluene and acetylated with 1:1
Ac2O–pyridine (2 mL) with a catalytic
amount of DMAP at rt overnight. The acety-
L
-
D
D
D
A soln of compound 24 (25 mg, 0.017 mmol)
and 10% Pd–C (25 mg) in 1:4 MeOH–

574
J. Xia et al. / Carbohydrate Research 329 (2000) 561–577
J1,2 3.0 Hz, H-1V), 4.88–4.78 (dd, 1 H, H-5V),
4.78–4.60 (d, 1 H, J1,2 3.0 Hz, H-1I), 4.60–
4.52 (d, 1 H, J1,2 8.8 Hz, H-1II), 4.52–4.46 (d,
1 H, J1,2 7.8 Hz, H-1IV), 4.46–4.44 (d, 1 H, J1,2
7.6 Hz, H-1II), 4.36–4.28 (dd, 1 H, H-2I),
4.28–4.24 (d, J 2.8 Hz, H-4I), 4.22–4.18 (d, 2
H), 4.14–3.84 (m, 11 H, H-5I, H-4III, H-4II,
H-2III, H-4IV, H-3III), 3.84–3.46 (m, 12 H,
H-4V, H-6aI, H-3IV, H-3II, H-5III, H-2IV, H-
2II), 3.36 (s, 3 H, OCH3), 2.13 (s, 3 H, Ac),
2.02 (s, 3 H, Ac), 1.20 (d, 3 H, J 6.4 Hz, CHV3 );
13C NMR (D2O, 100.6 MHz): l 173.50 (CꢀO),
173.20 (CꢀO), 103.40, 100.45, 100.22, 97.45,
97.20, 76.48, 74.45, 73.90, 73.83, 72.80, 71.55,
71.53, 71.50, 70.98, 70.00, 69.53 (2 C), 68.54,
68.52, 67.90, 67.40, 67.30, 66.75, 66.25, 65.63,
60.50, 58.90, 54.98, 54.00, 47.50, 21.25 (Ac),
21.00 (Ac), 14.25 (CHV3 ); FABMS (m/z) (posi-
tive ion mode): Calcd for C35H59N2NaO28S:
1012; found: 1034.0 [M+ +Na], 1012 [M+].
CH2Cl2 (3 mL) was stirred for 2.5 h at rt
under a H2 atmosphere. The solids were
filtered off, and the organic layer was concd.
The crude residue was treated for 6–9 h at
0–25 °C with the SO3·pyridine complex (10
mg) in pyridine (2 mL), then quenched with
MeOH and treated with Amberlite IR 120
(Na+) cation-exchange resin in MeOH at rt
for 1.5 h. The filtrate was concd and purified
by a short column of silica gel eluted with 10:1
CH2Cl2–MeOH to give pure compound 26
(25 mg, 97% in two steps) as an amorphous
1
solid. H NMR (CD3OD, 600 MHz): l 5.48–
5.40 (m, 3 H), 5.40–5.30 (m, 2 H), 5.28–5.20
(dd, 1 H), 5.16–4.4.92 (m, 5 H), 4.75–4.70 (d,
1 H), 4.70–4.60 (m, 3 H), 4.48–4.32 (m, 2 H),
4.32–4.28 (m, 2 H), 4.24–4.04 (m, 4 H), 4.04–
3.92 (m, 3 H), 3.92–3.60 (m, 6 H), 3.52–3.44
(m, 1 H), 3.38 (s, 3 H, OCH3), 2.17 (s, 3 H,
Ac), 2.16 (s, 3 H, Ac), 2.15 (s, 3 H, Ac), 2.14
(s, 3 H, Ac), 2.09 (s, 6 H, 2 Ac), 2.08 (2s, 6 H,
2 Ac), 2.04 (s, 3 H, Ac), 2.00 (s, 3 H, Ac), 1.96
(2s, 6 H, 2 Ac), 1.93 (s, 3 H, Ac), 1.90 (s, 3 H,
Ac), 1.20 (d, 3 H, J 6.4 Hz, CH3); 13C NMR
(CD3OD, 100.6 MHz): l 173.38 (CꢀO), 173.21
(CꢀO), 172.57 (CꢀO), 172.55 (CꢀO), 172.33
(CꢀO), 172.23 (CꢀO), 172.11 (CꢀO), 171.60
(CꢀO), 171.16 (CꢀO), 103.03, 102.36, 101.79,
100.14, 96.99, 76.66, 75.48, 74.43, 74.15, 73.01,
72.60, 72.42, 72.32, 72.26, 72.02, 70.60, 70.52,
70.34, 69.82, 69.63, 69.38, 68.81, 68.70, 66.07,
65.45, 63.56, 62.52, 56.87, 56.14, 55.69, 23.65
(NAc), 23.22 (NAc), 21.47 (Ac), 21.35 (Ac),
21.30 (Ac), 21.21 (Ac), 21.17 (Ac), 21.07 (Ac),
20.99 (Ac), 20.96 (Ac), 20.93 (Ac), 16.70
(CH3).
Methyl
(2,3,4-tri-O-acetyl-6-O-trimethyl-
-galactopyranosyl)-(1“4)-[(2,3,4-
acetyl-i-
D
tri-O-benzyl-h- -fucopyranosyl)-(1“3)]-2-
L
deoxy-6-O-naphthylmethyl-2-phthalimido-i-
glucopyranosyl)-(1“6)-[(2,3,4-tri-O-benzoyl-
i- -galactopyranosyl)-(1“3)]-2-acetamido-
2-deoxy-h- -galactopyranoside (29).—A soln
D
-
D
D
of trisaccharide donor 16 (722 mg, 0.54
mmol), disaccharide acceptor 27 (460 mg, 0.50
mmol), and N-iodosuccinimide (NIS, 365 mg,
0.62 mmol) in dry CH2Cl2 (12 mL) containing
,
4 A MS (10 g) was stirred for 2 h at −65 to
−70 °C under a N2 atmosphere. Trifluoro-
methanesulfonic acid (TfOH) (60 mL) in
dry CH2Cl2 (2 mL) was then added dropwise,
and the mixture was stirred for 2 h at the
same temperature. The mixture was neutral-
ized with satd aq NaHCO3. The solids were
filtered off, and the organic layer was washed
with satd NaHCO3 soln, 10% Na2S2O3, and
water. The organic extract was dried (Na2SO4)
and concd under reduced pressure. The crude
mixture was applied to a column of silica gel
and eluted with 1:1 hexane–EtOAc to give
pure compound 28 (490 mg, 46%) as an amor-
phous solid. Compound 28 (490 mg, 0.23
mmol), 2,6-lutidine (10 mL) and thiourea (52
mg) in 1:1 EtOH–CH2Cl2 (30 mL) were
stirred overnight at 60–65 °C. The reaction
mixture was then concd to give a crude
product, which was applied to a column of
Methyl (i -
[(h- -fucopyranosyl)-(1“3)]-(2-acetamido-2-
deoxy-i- -glucopyranosyl)-(1“6)-[(6-O-sulfo-
i- -galactopyranosyl)-(1“3)]-2-acetamido-2-
deoxy-h- -galactopyranoside sodium salt (1).
D
- galactopyranosyl) - (1“4)-
L
D
D
D
—Sodium methoxide (80 mL of a 1 M soln)
was added to a soln of compound 26 (25 mg,
0.017 mmol) in 1:1 MeOH–water (3 mL). The
reaction mixture was stirred for 24 h at rt and
concd. The crude product was applied to a
short column of silica gel and eluted with 3:1:1
n-C3H7OH–HOAc–water to give pure com-
pound 1 (8 mg, 48%) as an amorphous solid.
1
Rf 0.26 (3:1:1 n-C3H7OH–HOAc–water); H
NMR (D2O, 600 MHz): l 5.16–5.08 (d, 1 H,

J. Xia et al. / Carbohydrate Research 329 (2000) 561–577
575
(35 mg, 0.23 mmol) was added to a cold (ice
bath) soln of compound 29 (300 mg, 0.15
mmol) in dry pyridine (2 mL). The mixture
was stirred for 18–24 h at the same tempera-
ture then quenched with MeOH and treated in
MeOH with Amberlite IR 120 (Na+) cation-
exchange resin. The filtrate was concd to give
a crude residue, which was applied to a short
column of silica gel and eluted with 30:1
CH2Cl2–MeOH to give pure compound 30
(260 mg, 82%) as an amorphous solid. [h]D
silica gel and eluted with 1:3 hexane–EtOAc
to give pure compound 29 (390 mg, 84%) as
an amorphous solid. Rf 0.47 (1:3 hexane —
EtOAc); [h]D +70.8° (c 0.6, CHCl3); 1H
NMR (CDCl3, 600 MHz): l 8.10–7.82 (m, 10
H, ArH), 7.80–7.77 (m, 3 H, ArH), 7.60–7.50
(m, 2 H, ArH), 7.50–7.30 (m, 10 H, ArH),
7.30–7.10 (m, 15 H, ArH), 7.00–6.90 (m, 1 H,
ArH), 5.75–5.15 (m, 2 H, H-4II, H-2II), 5.60–
5.50 (dd, 1 H), 5.30–5.20 (d, 1 H), 5.15–5.05
(m, 2 H, H-4V, H-1III, J1,2 8.6 Hz), 5.01–4.98
(dd, 1 H, H-2IV), 4.96–4.92 (d, 2 H, Jgem 12.6
Hz, 2 OCHAr), 4.90–4.84 (d, 1 H, J1,2 7.8 Hz,
H-1II), 4.84–4.75 (m, 3 H, OCHAr, H-1I, H-
3IV), 4.75–4.64 (m, 3 H, H-1IV, H-3III,
OCHAr), 4.64–4.60 (m, 2 H, H-5V), 4.60–
4.52 (d, 2 H, Jgem 12.4 Hz, 2 OCHPh), 4.40–
4.32 (m, 2 H, H-2III), 4.32–4.24 (dd, 2 H,
OCH2Ph), 4.20–4.12 (t, 1 H, H-4III), 4.12–
4.04 (m, 2 H), 4.04–3.96 (t, 1 H, H-5II),
3.92–3.84 (m, 2 H, H-3I), 3.84–3.76 (m, 1 H),
3.76–3.69 (dd, 1 H, H-2I), 3.69–3.48 (m, 7 H,
H-5III), 3.12–3.04 (t, 1 H), 2.90–2.84 (t, 1 H),
2.84 (s, 3 H, OCH3), 1.92 (s, 3 H, Ac), 1.84 (s,
3 H, Ac), 1.80 (s, 3 H, Ac), 1.79 (s, 3 H, Ac),
1.20 (d, 3 H, J 6.4 Hz, CHV3 ), 1.12 (s, 9 H,
t-Bu); 13C NMR (CDCl3, 100.6 MHz): l
177.85 (CꢀO), 170.20 (CꢀO), 170.00 (CꢀO),
169.90 (CꢀO), 168.90 (CꢀO), 166.85 (CꢀO),
165.78 (CꢀO), 165.00 (CꢀO), 138.85, 138.45,
135.10, 134.23, 134.00, 133.60, 133.48, 133.41,
133.25, 130.27, 130.00, 129.86, 128.89, 128.76,
128.47, 128.40, 128.30, 128.24, 128.18, 128.16,
120.05, 127.82, 127.60, 127.36, 127.25, 126.60,
126.30, 126.04, 123.60, 102.30, 99.86, 99.00,
98.50, 97.80, 80.00, 79.86, 75.66, 75.45, 75.20,
75.00, 74.50, 74.20, 73.21, 72.55, 71.89, 71.40,
70.50, 70.34, 69.30, 69.20, 68.80, 68.50, 68.00,
67.78, 67.00, 66.70, 61.00, 60.30, 56.50, 54.80,
48.00, 38.85, 27.50, 22.50 (Ac), 20.58 (3 Ac),
16.85 (CH3); Anal Calcd for C105H110N2O33:
C, 65.60; H, 5.65; N, 1.43. Found: C, 65.35;
H, 5.81; N, 1.64.
1
+38° (c 0.2, CHCl3); H NMR (CD3OD, 600
MHz): l 8.15–7.86 (m, 10 H, ArH), 7.90–7.78
(m, 2 H, ArH), 7.60–7.50 (m, 3 H, ArH),
7.50–7.40 (m, 10 H, ArH), 7.30–7.15 (m, 14
H, ArH), 7.00–6.90 (m, 2 H, ArH), 5.33–5.26
(m, 2 H, H-4II, H-2II), 5.70–5.55 (dd, 1 H),
5.25–5.20 (d, 1 H), 5.15–5.08 (m, 2 H, H-4IV,
H-1III, J1,2 8.6 Hz), 5.05–4.98 (dd, 1 H, H-2IV),
4.96–4.94 (d, 1 H, Jgem 12.6 Hz, OCHAr),
4.90–4.84 (d, 1 H, J1,2 7.8 Hz, H-1II), 4.84–
4.77 (m, 4 H, 2 OCHAr, H-1I, H-3IV), 4.76–
4.68 (m, 3 H, H-1IV, H-3III, OCHAr),
4.64–4.60 (m, 2 H, H-5V), 4.60–4.52 (d, 2 H,
Jgem 12.4 Hz, 2 OCHPh), 4.48–4.32 (m, 2 H,
H-2III), 4.32–4.24 (dd, 2 H, OCH2Ph), 4.20–
4.12 (t, 1 H, H-4V), 4.12–4.04 (m, 2 H),
4.04–3.96 (t, 1 H), 3.92–3.84 (m, 2 H, H-3I),
3.84–3.76 (m, 1 H), 3.76–3.69 (dd, 1 H, H-2I),
3.69–3.48 (m, 7 H, H-5III), 3.12–3.04 (m, 1
H), 2.90–2.84 (m, 1 H), 2.84 (s, 3 H, OCH3),
1.92 (s, 3 H, Ac), 1.88 (s, 3 H, Ac), 1.87 (s, 3
H, Ac), 1.80 (s, 3 H, Ac), 1.21 (d, 3 H, J 6.4
Hz, CHV3 ), 1.12 (s, 9 H, t-Bu); 13C NMR
(CDCl3, 100.6 MHz): l 177.86 (CꢀO), 170.26
(CꢀO), 170.05 (CꢀO), 169.98 (CꢀO), 168.93
(CꢀO), 166.88 (CꢀO), 165.79 (CꢀO), 165.00
(CꢀO), 138.95, 138.45, 135.10, 134.23, 134.08,
133.60, 133.48, 133.41, 133.26, 130.27, 130.05,
129.85, 128.86, 128.76, 128.37, 128.30, 128.28,
128.24, 128.15, 128.11, 120.05, 127.88, 127.65,
127.36, 127.35, 126.80, 126.37, 126.24, 123.60,
102.30, 99.86, 99.00, 98.50, 97.80, 80.00, 79.86,
75.66, 75.45, 75.20, 75.08, 74.50, 74.20, 73.21,
72.55, 71.89, 71.46, 70.55, 70.34, 69.34, 69.20,
68.85, 68.50, 68.00, 67.86, 67.80, 66.70, 61.06,
60.30, 56.50, 54.20, 48.25, 38.85, 27.50, 22.60
(Ac), 20.59 (3 Ac), 16.45 (CH3).
Methyl
(2,3,4-tri-O-acetyl-6-O-trimethyl-
-galactopyranosyl)-(1“4)-[(2,3,4-
acetyl-i-
D
tri-O-benzyl-h- -fucopyranosyl)-(1“3)]-2-
L
deoxy-6-O-naphthylmethyl-2-phthalimido-i-
glucopyranosyl)-(1“6)-[(2,3,4-tri-O-benzoyl-
6-O-sulfo-i- -galactopyranosyl)-(1“3)]-2-
acetamido - 2 - deoxy - h - - galactopyranoside
sodium salt (30).—Sulfur trioxide–pyridine
D
-
D
Methyl
acetyl-i-
(2,3,4-tri-O-acetyl-6-O-trimethyl-
-galactopyranosyl)-(1“4)-[(2,3,4-
D
D
tri-O-benzyl-h- -fucopyranosyl)-(1“3)]-2-
L

576
J. Xia et al. / Carbohydrate Research 329 (2000) 561–577
deoxy-2-phthalimido-6-O-sulfo-i-
ranosyl) - (1“6) - [(2,3,4 - tri - O - benzoyl - 6 - O-
sulfo-i- -galactopyranosyl)-(1“3)]-2-aceta-
mido-2-deoxy-h- -galactopyranoside disodium
D
-glucopy-
133.61, 133.48, 133.41, 133.25, 130.27, 130.01,
129.86, 128.89, 128.76, 128.47, 128.40, 128.32,
128.24, 128.18, 128.16, 128.05, 127.82, 127.62,
127.36, 127.25, 126.63, 126.31, 126.04, 123.61,
102.35, 99.86, 99.08, 98.52, 97.81, 80.08, 79.86,
75.66, 75.45, 75.21, 75.05, 74.52, 74.23, 73.21,
72.55, 71.89, 71.40, 70.50, 70.34, 69.35, 69.20,
68.83, 68.57, 68.08, 67.86, 67.81, 66.75, 61.08,
60.31, 56.56, 54.87, 48.50, 38.85, 27.51, 22.53
(Ac), 20.58 (3 Ac), 16.35 (CH3).
D
D
salt (32).—To a soln of compound 30 (235
mg, 0.12 mmol) in 4:1:trace CH2Cl2 –MeOH–
water (10 mL) was added DDQ (40 mg). After
stirring for 3.5 h at rt, an additional portion
of DDQ (20 mg) was added, and the mixture
was stirred at the same temperature for a total
of 16 h. The mixture was concd under reduced
pressure to give a crude residue, which was
dissolved in CH2Cl2 (100 mL) and washed
with satd NaHCO3 soln (3×100 mL). The
organic extract was dried over Na2SO4 and
concd. The crude product was applied to a
column of silica gel and eluted with 20:1
CH2Cl2–MeOH to give pure compound 31
(120 mg, yield: 51%) as an amorphous solid.
Compound 31 was treated with SO3·pyridine
for 12 h in dry pyridine at 0–5 °C, then
purified by a short column of silica gel that
was eluted with 15:1 CH2Cl2 –MeOH to give
pure compound 32 (79%) as an amorphous
Methyl (i -
[(h- -fucopyranosyl)-(1“3)]-2-acetamido-2-
deoxy-6-O-sulfo-i- -glucopyranosyl)-(1“6)-
[(6-O-sulfo-i- -galactopyranosyl)-(1“3)]-2-
acetamido-2-deoxy-h- -galactopyranoside di-
D
- galactopyranosyl) - (1“4)-
L
D
D
D
sodium salt (2).—A soln of compound 32 (90
mg) and 10% Pd–C (200 mg) in 3:1 CH2Cl2–
MeOH (6 mL) was stirred overnight at rt
under a H2 atmosphere. The filtrate was concd
under reduced pressure to give a crude residue
that was treated for 4–6
h
in 1:5
NH2NH2·H2O–MeOH (12 mL) at 80–85 °C.
The reaction mixture was then concd and
treated overnight with 1:1 Ac2O–pyridine (6
mL) in the presence of a catalytic amount of
DMAP at rt. The mixture was concd and
purified by a short column of silica gel and
eluted with 6:1 CH2Cl2–MeOH to give pure
compound 33. Compound 33 was dissolved in
1:1 MeOH–water (3 mL), treated for 24 h
with 1 M CH3ONa (80 mL) at rt, and then
concd. The crude residue was applied to a
short column of silica gel and eluted with 3:1:1
n-C3H7OH–HOAc–water to give compound
2 (6 mg, 26%) as an amorphous solid. Rf 0.16
1
solid. [h]D +59° (c 0.5, CHCl3); H NMR
(D2O, 600 MHz): l 8.10–7.82 (m, 10 H,
ArH), 7.80–7.77 (m, 3 H, ArH), 7.70–7.59
(m, 2 H, ArH), 7.56–7.30 (m, 6 H, ArH),
7.30–7.15 (m, 12 H, ArH), 7.00–6.90 (m, 1 H,
ArH), 5.70–5.65 (m, 2 H, H-4II, H-2II), 5.60–
5.50 (dd, 1 H, H-3II), 5.30–5.20 (d, 1 H),
5.15–5.05 (m, 2 H, H-4IV, H-1III, J1,2 8.6 Hz),
5.01–4.98 (dd, 1 H, H-2IV), 4.96–4.92 (d, 1 H,
Jgem 12.6 Hz, OCHAr), 4.90–4.84 (d, 1 H, J1,2
7.8 Hz, H-1II), 4.84–4.75 (m, 3 H, OCHAr,
H-1I, H-3IV), 4.75–4.64 (m, 3 H, H-1IV, H-3III,
OCHAr), 4.64–4.60 (m, 2 H, H-5V), 4.60–
4.52 (d, 1 H, Jgem 12.4 Hz, OCHPh), 4.04–
4.32 (m, 2 H, H-2III), 4.32–4.24 (dd, 2 H,
OCH2Ph), 4.20–4.12 (t, 1 H, H-4III), 4.12–
4.04 (m, 2 H), 4.04–3.96 (t, H-5II), 3.92–3.84
(m, 2 H, H-3I), 3.84–3.76 (m, 1 H), 3.76–3.69
(dd, 1 H, H-2I), 3.69–3.48 (m, 7 H, H-5III),
3.12–3.04 (t, 1 H), 2.90–2.84 (t, 1 H), 2.84 (s,
3 H, OCH3), 1.92 (s, 3 H, Ac), 1.84 (s, 3 H,
Ac), 1.80 (s, 3 H, Ac), 1.79 (s, 3 H, Ac), 1.20
(d, 3 H, J 6.4 Hz, CH3V), 1.12 (s, 9 H, t-Bu);
13C NMR (CDCl3, 100.6 MHz): l 176.85
(CꢀO), 170.25 (CꢀO), 170.00 (CꢀO), 169.93
(CꢀO), 168.91 (CꢀO), 166.85 (CꢀO), 165.78
(CꢀO), 165.01 (CꢀO), 135.13, 134.23, 134.00,
1
(3:1:1 n-C3H7OH–HOAc–water); H NMR
(D2O, 600 MHz): l 5.16–5.10 (d, 1 H, J1,2 3.5
Hz, H-1V), 4.90–4.79 (dd, 1 H, H-5V), 4.78–
4.60 (d, 1 H, J1,2 3.0 Hz, H-1I), 4.64–4.60 (d,
1 H, J1,2 8.8 Hz, H-1III), 4.60–4.58 (d, 1 H, J1,2
7.8 Hz, H-1IV), 4.46–4.44 (d, 1 H, J1,2 7.6 Hz,
H-1II), 4.44–4.40 (d 1 H, J 2.8 Hz, H-4IV),
4.40–4.30 (dd, 1 H, H-2I), 4.30–4.28 (dd, 1 H,
J 2.8 Hz, H-4I), 4.22–4.18 (m, 2 H, H-6bII,
H-5I), 4.14–3.80 (m, 11 H, H-3I, H-6bI, H-4II,
H-2III, H-3V, H-6aI, H-5IV, H-4V), 3.80–3.46
(m, 12 H, H-5III, H-2V, H-5II, H-3IV, H-3II,
H-4III, H-2IV, H-2II), 3.36 (s, 3 H, OCH3), 2.13
(s, 3 H, Ac), 2.02 (s, 3 H, Ac), 1.20 (d, 3 H, J
6.4 Hz, CHV3 ); 13C NMR (D2O, 100.6 MHz): l
173.50 (CꢀO), 173.20 (CꢀO), 104.50, 102.30,

J. Xia et al. / Carbohydrate Research 329 (2000) 561–577
577
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101.60, 98.60, 98.40, 77.52, 75.20, 75.00, 73.30,
72.90, 72.75, 72.65 (2 C), 72.25, 71.50, 71.25,
70.60, 69.50 (2 C), 69.25, 68.60, 68.50, 68.00,
67.50, 67.00, 66.30, 62.65, 56.00, 55.28, 48.90,
23.00 (Ac), 22.50 (Ac), 15.50 (CHV3 ); FABMS
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Acknowledgements
The authors thank Dr S. Dutta for mass
spectrometry analyses. This work was sup-
ported by Grant No. CA 35329 and in part by
Grant No. P30CA16056, both awarded by
National Cancer Institute.
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