Synthesis of a neoglycoprotein containing the Lewis X analogous trisaccharide β-D-GalpNAc-(1 →4)[α-L-Fucp-(1→3)]-β-D-GlcpNAc

Article abstract of DOI:10.1016/S0008-6215(98)00082-2

The trisaccharide allyl glycoside 36 and related disaccharide part structures have been prepared using the 2-trichloroacetamido-2-deoxy-α-D-galactopyranosyl trichloroacetimidate derivative 9 as glycosyl donor under promotion with TMSOTf or Sn(OTf)₂

Full text of DOI:10.1016/S0008-6215(98)00082-2

Carbohydrate Research 308 (1998) 259±273
Synthesis of a neoglycoprotein containing the
Lewis X analogous trisaccharide ꢀ-d-GalpNAc-
(1!4)[ꢁ-l-Fucp-(1!3)]-ꢀ-d-GlcpNAc
Johannes Bartek, Renate Muller, Paul Kosma*
Institute of Chemistry, University of Agricultural Sciences Vienna, Muthgasse 18, A-1190 Vienna,
Austria
Received 10 December 1997; accepted 10 March 1998
Abstract
The trisaccharide allyl glycoside 36 and related disaccharide part structures have been prepared
using the 2-trichloroacetamido-2-deoxy-ꢁ-d-galactopyranosyl trichloroacetimidate derivative 9 as
glycosyl donor under promotion with TMSOTf or Sn(OTf)2, respectively, to produce the ꢀ-(1!4)
linkage to suitably protected glucosamine derivatives in fair yields. Fucosylation was e€ected
employing the ethyl 1-thio glycosyl donor 20 in the presence of IDCP. Deprotection of the inter-
mediates a€orded the disaccharide allyl glycosides ꢀ-d-GalpNAc-(1!4)-ꢀ-d-GlcpNAc 13, ꢀ-d-
GalpNClAc-(1!4)-ꢀ-d-GlcpNAc 14, ꢁ-l-Fucp-(1!3)-ꢀ-d-GlcpNAc 24, ꢁ-l-Fucp-(1!4)-ꢀ-d-
GlcpNAc 31 and the branched trisaccharide allyl glycoside ꢀ-d-GalpNAc-(1!4)[ꢁ-l-Fucp-(1!3)]-
ꢀ-d-GlcpNAc 36. The trisaccharide which corresponds to a structural motif occurring in N-gly-
coprotein glycans from human urokinase, human recombinant protein C, phospholipase A2 as
well as O-glycans, was converted into a neoglycoprotein following introduction of a cysteamine-
derived spacer group and subsequent activation with thiophosgene. # 1998 Elsevier Science Ltd.
All rights reserved
Keywords: Lewis X; N-Acetyl-d-galactosamine; Oligosaccharide synthesis
1. Introduction
Bothrops moojeni toxins [4]. The presence of this
trisaccharide has furthermore been demonstrated
for egg jelly coats from Axolotl maculatum [5] and
N-glycans of Schistosomas mansoni [6]. Moreover,
it constitutes an important determinant in the N-
glycans of bovine pro-opiomelanocortin [7],
human urokinase [8] and recombinant human pro-
tein C expressed in kidney 293 cells [9]. It has been
postulated that the trisaccharide unit e€ectively
inhibits selectin-mediated cell adhesion and thus
seems to be a more potent ligand for E-selectin
than sialyl Lewis X [10].
The terminal trisaccharide determinant ꢀ-d-
GalpNAc-(1!4)[ꢁ-l-Fucp-(1!3)]-ꢀ-d-GlcpNAc
has been found in N- and O-glycans from various
organisms. Thus, it has been isolated from aller-
genic H-antigens of sea squirt [1] and described as
a structural element occurring in phospholipase A2
and hyaluronidase of the honey bee venom [2,3]
and in a serine protease from the snake venom of
* Corresponding author.
0008-6215/98/$19.00 # 1998 Elsevier Science Ltd. All rights reserved
PII S0008-6215(98)00082-2

260
J. Bartek et al./Carbohydrate Research 308 (1998) 259±273
Previously, the trisaccharide has been prepared
(!4) and subsequent oxidative removal of the 4-O-
p-methoxybenzyl group, which was accomplished
in 72% yield by treatment with ceric ammonium
nitrate and sonication. As the glycosyl donor for
the galactosamine unit, the 2-deoxy-2-trichloro-
acetamido trichloroacetimidate derivative 9 was
elaborated, since related glucosyl donors have been
used for ecient synthesis of chitobiosyl units [14].
Starting from previously described [15] N-p-meth-
oxybenzylidene derivative 6, amine 7 was obtained
by acidic hydrolysis with 1 M HCl in 86% yield,
which was then acylated with trichloroacetyl
chloride±triethylamine in dichloromethane to give
compound 8 in 99% yield. The anomeric acetate
was selectively removed with hydrazine acetate [16]
and the resulting reducing galactosamine derivative
was converted into the ꢁ-trichloroacetimidate 9 by
reaction with trichloroacetonitrile and 1,8-diazabi-
cyclo[5.4.0]undec-7-ene (DBU) [17] in dichloro-
methane in 78% yield. Trimethylsilyl tri¯uoro-
methanesulfonate-promoted coupling of the tri-
chloroacetimidate donor 9 with acceptor 5 a€orded
the ꢀ-(1!4)-linked crystalline disaccharide deriva-
tive 10 in 37% yield. The structural assignment was
evident from the value of the coupling constant
as the methyl glycoside by a chemo-enzymatic
approach employing human milk fucosyltransfer-
ase [11]. For further biological studies, we have set
out to prepare neoglycoproteins containing this
trisaccharide determinant. Here, we report on the
synthesis of the corresponding allyl glycoside
which may be used for the introduction of a
spacer function and the design of glycopolymers.
A 2-deoxy-2-trichloro-acetamido-galactopyranosyl
donor was elaborated to provide access for addi-
tional chemical transformations at the amino
group of the terminal galactosamine unit.
2. Results and discussion
For the synthesis of the 20-amino-20-deoxy-lacto-
samine derivatives 13 and 14, known [12] allyl
2-acetamido-2-deoxy-4,6-O-p-methoxybenzylidene-
ꢀ-d-glucopyranoside 1 was subjected to reductive
ring-opening [13] in N,N-dimethylformamide in
the presence of sodium cyanoborohydride±tri-
¯uoroacetic acid which proceeded with low selec-
tivity to give the 4-O- and 6-O-p-methoxybenzyl
ether derivatives 2 and 3 in a 1:1.1 ratio and 77%
yield (Scheme 1). The isomers were separated by
chromatography and elaborated into suitable gly-
cosyl acceptors. Thus, compound 2 was converted
into the 3,6-di-O-acetyl derivative 5 via acetylation
0
0
J1 ,2 (8.3 Hz) of the galactosamine unit and the
chemical shift of H-4 (ꢂ 3.78) of the glucosamine
residue. Promotion of the glycosidation reaction by
either stannous tri¯uoromethanesulfonate or zinc
tri¯uoromethanesulfonate [18] resulted in even
Scheme 1.

J. Bartek et al./Carbohydrate Research 308 (1998) 259±273
261
lower yields (<20%) of the disaccharide product.
Reduction [19] of the N-trichloroacetamido group
of 10 could be achieved in the presence of sodium
borohydride in ethanol to give after N-acetylation
the disaccharide derivative 11 in 68% yield. More-
over, partial reduction of the trichloroacetamide
was carried out in 78% yield using zinc dust in
acetic acid at 110 ꢀC to furnish the N-chloro-
acetamido derivative 12.
Deprotection of both protected disaccharide
derivatives 11 and 12 with sodium methoxide in
methanol proceeded in near quantitative yield to
give the N-acetyl and N-chloroacetyl compounds
13 and 14, respectively. Due to the down®eld-shift
of the chloromethylene protons, the remaining
methyl group of the reducing glucosamine unit
and the chloroacetamido group in compound 14
are amenable for selective NOE experiments in
conformational studies.
Since the anomeric allyl group was chosen for
the introduction of the spacer moiety, the use of
benzyl-protected fucopyranosyl donors was not
feasible. Instead, the previously described [20] tri-
O-benzoyl-l-fucopyranosyl bromide 15 was
employed (Scheme 2). Glycosylation of either N-
acetyl or N-phthaloyl protected 4,6-O-p-methoxy-
benzylidene derivative 1 or 17 [21] was promoted
by mercuric cyanide±mercuric bromide to give the
ꢀ-(1!3)-linked disaccharide 16 in 75% yield and a
1:1 anomeric mixture of the N-phthalimido deri-
vatives 18 and 19 in 38% yield, respectively, in
accordance with the results of Nifant'ev et al. [20].
By using the ethyl 1-thio glycoside 20 [22] under
promotion of iodonium di-sym-collidinium per-
chlorate (IDCP) [23], the ꢁ-(1!3)-linked disa-
ccharide derivative 21 was formed with greatly
enhanced selectivity (ꢁ=ꢀ ratio 12:1) and in fair
yield (49%). Oxidative removal of the 4,6-O-p-
methoxybenzylidene group with ceric ammonium
nitrate furnished the 4,6-diol derivative 22, whereas
treatment of 21 with 2,3-dichloro-5,6-dicyano-1,4-
benzoquinone (DDQ) lead to simultaneous clea-
vage of the 20-O-p-methoxybenzyl ether and the
30,40-O-isopropylidene group, respectively, to give,
after O-acetylation, the acetylated disaccharide
derivative 23 in 92% overall yield. Deprotection in
the presence of sodium methoxide a€orded the
disaccharide ꢁ-l-Fucp-(1!3)-ꢀ-d-GlcpNAc allyl
glycoside 24. (Scheme 2).
Since the reductive ring opening of the 4,6-O-p-
methoxybenzylidene group of 21 in the presence of
sodium cyanoborohydride±tri¯uoroacetic acid met
with diculties due to the acid lability of the
fucosidic bond, the disaccharide acceptor 25 was
prepared via regioselective glycosylation of the
3,4-diol compound 3 (Scheme 3). Iodonium
Scheme 2.

262
J. Bartek et al./Carbohydrate Research 308 (1998) 259±273
Scheme 3.
di-sym-collidinium perchlorate-promoted coupling
of 3 with 1.1 equivalents of the ethyl 1-thioglyco-
side donor 20 in 1:5 N,N-dimethylformamide±
dichloromethane a€orded a mixture of three di-
saccharides which was separated by column chro-
matography to give 32% of the ꢁ-(1!3)-linked
disaccharide 25, a minor portion (4%) of the corre-
sponding ꢀ-isomer 26 and the ꢁ-(1!4)-linked
disaccharide derivative 28 in 11% yield. Whereas
the anomeric con®gurations were deduced from the
(anomeric bromide or trichloroacetimidate) in the
presence of silver tri¯uoromethanesulfonate or
borontri¯uoride etherate, respectively, under stan-
dard and ``inverse conditions'' [25] did not lead to
signi®cant product formation. Employing the N-
trichloroacetamido donor 9 under promotion of
trimethylsilyl tri¯uoromethanesulfonate, silver tri-
¯uoromethanesulfonate, zinc tri¯uoromethane-
sulfonate, stannic chloride, or stannous chloride
resulted in the formation of by-products or hydro-
lysis of the protecting groups of the acceptor disa-
ccharide. Finally, stannous tri¯uoromethane-
sulfonate-mediated glycosylation [26] under care-
fully controlled conditions proved to be e€ective.
Thus, using 1.5 equivalents of 9, 1.3 equivalents of
promoter and 1.1 equivalents of N,N,N0,N0-tetra-
methyl urea in dichloromethane±toluene for 1.5 h
at room temperature furnished the trichloro-
methyloxazoline derivative 32 (36%), the ꢀ-(1!4)-
linked trisaccharide derivative 33 (41%) and
unreacted starting material 25 (26%), which were
separated by column chromatography. The ꢀ-
anomeric con®guration of 33 was evident from the
0
values of the proton coupling constants J1 ,2
0
(3.5 Hz for 25 and 28, 8.3 Hz for 26), the attach-
ment site was assigned after subsequent O-acetyla-
tion to give 27 and 29, respectively. Upon
acetylation, H-4 of compound 27 experienced a
down®eld shift (to 4.88 ppm), whereas H-3 of 29 was
shifted to 5.08 ppm. Compound 28, corresponding
to a synthetic part structure of the Lewis A blood-
group antigenic determinant [24] was deprotected
via hydrolysis of the blocking groups in aqueous
90% tri¯uoroacetic acid followed by reacetylation
to give 30 in 83% yield. Zemplen de-O-acetylation
of 30 gave the disaccharide ꢁ-l-Fucp-(1!4)-ꢀ-d-
GlcpNAc allyl glycoside 31.
00 00
value J1 ,2 (9.0 Hz) indicating a trans-diaxial
Various attempts to introduce the ꢀ-(1!4)-
linked galactosamine residues by coupling of the
disaccharide acceptor 25 with 2-acetamido-3,4,6-
tri-O-acetyl-2-deoxy-ꢁ-d-galactopyranosyl donors
arrangement of the respective protons. Deblocking
was achieved by ®rst removing the p-methoxy-
benzyl ether groups of 33 by treatment with 2,3-
dichloro-5,6-dicyano-1,4-benzoquinone followed

J. Bartek et al./Carbohydrate Research 308 (1998) 259±273
263
by O-acetylationÐto stabilize the fucosidic link-
ageÐwhich a€orded 34 in 96% yield. Hydrolysis
of the isopropylidene acetal group was accom-
plished in the presence of aqueous 90% tri-
protein ratio of the BSA conjugate 38 based on
protein [29] and aminosugar analysis [30] was 6:1
mol/mol.
ꢀ
¯uoroacetic acid at 20 C, followed by reduction
of the trichloroacetamido group with sodium cya-
3. Experimental
ꢀ
noborohydride at 60 C. Subsequent O-acetylation
furnished the per-O-acetyl trisaccharide compound
35 in 60% yield (3 steps). Removal of the O-acetyl
groups by treatment with sodium methoxide a€or-
ded the target trisaccharide derivative 36 in near
quantitative yield. Due to the branched substitu-
tion pattern, the 13C NMR signals of C-4 and C-3
of the glucosamine unit of 36 displayed no sig-
ni®cant glycosylation shifts in contrast to the
respective signals observed for the disaccharide
part structures (Table 1).
Finally, the allyl glycoside was transformed in
96% yield into the 3-(2-aminoethylthio)propyl
spacer [27] derivative 37 by radical addition of
cysteamine hydrochloride under UV irradiation
[28]. The spacer compound 37 was isolated as the
acetate, then activated with thiophosgene and
coupled to the "-amino groups of lysine residues
of bovine serum albumin (BSA). The ligand/
General Methods.ÐMelting points were deter-
mined with a Ko¯er hot stage and are uncorrected.
Optical rotations were measured with a Perkin±
20
Elmer 243 B polarimeter; [ꢁ]d values are given in
1
1
units of 10 1 degcm2 g . H NMR spectra were
recorded at 297 K with a Bruker AC 300F instru-
ment operating at 300 MHz for 1H using CDCl3 as
solvent and tetramethylsilane as internal standard,
1
unless stated otherwise. 500 MHz H spectra were
recorded on a Bruker AMX instrument. Coupling
constants are given in Hz (®rst order values). 13C
NMR spectra were measured at 75.47 MHz and
referenced to 1,4-dioxane (ꢂ 67.40). Homo- and
heteronuclear 2D NMR spectroscopy was per-
formed with Bruker standard software. TLC was
performed on Merck precoated plates (5Â10 cm,
layer thickness 0.25 mm, Silica Gel 60F254); spots
were detected by spraying with anisaldehyde-H2SO4.
Table 1
13C NMR dataa for disaccharides 13, 14, 24, 31 and trisaccharide 36
ꢂ(ppm)
Residue
Carbon
13
14
24
31
36
ꢁ-l-Fucp
1
2
3
4
5
6
1
2
3
4
5
6
1
2
3
4
5
6
1
2
3
100.83
68.86
70.43
72.73
67.79
16.06
100.44
68.88
70.22
72.71
67.82
16.02
99.27
68.59
70.05
72.86
67.76
16.21
101.57
53.21
71.60
68.19
75.67
62.26
100.71
56.39
75.54
74.26
76.31
60.89
71.40
134.18
119.07
23.02
23.02
175.68
175.14
ꢀ-d-GalpNR
ꢀ-d-GlcpNAc
Aglycon
102.58
53.39
71.52
68.45
76.18
61.80
100.82
55.66
73.44
79.77
75.39
61.00
71.34
134.14
119.04
23.03
22.98
175.63
175.45
102.01
53.98
71.26
68.51
76.22
61.82
100.85
55.74
73.41
79.59
75.38
61.18
71.34
134.18
119.03
23.00
100.75
56.13
81.31
69.50
76.77
61.67
71.37
134.23
119.06
23.07
100.78
56.82
73.47
78.25
76.08
60.82
71.24
134.18
119.00
22.94
NHAc
CO
174.45
171.31
43.14
175.55
175.20
CH2Cl
a
Spectra (75.47 MHz) were recorded at 297 K and referenced to 1,4-dioxane (ꢂ 67.40).

264
J. Bartek et al./Carbohydrate Research 308 (1998) 259±273
For column chromatography, silica gel (0.040±
0.063 mm) was used. Concentration of solutions
was performed at reduced pressure at temperatures
OCH3), 3.67 (dd, 1 H, J2,3 10.0 Hz, H-2), 3.64 (dd,
1 H, J6b,5 5.8 Hz, H-6b), 3.43 (dd, 1 H, J3,4 8.5 Hz,
H-3), 3.39 (ddd, 1 H, J5,4 9.5 Hz, H-5), 3.34±3.28
(unresolved signal, 1 H, H-4), 1.96 (s, 3 H, Ac).
Anal. Calcd for C19H27NO7: C, 59.83; H, 7.13; N,
3.67. Found: C, 59.85; H, 7.27. N, 3.61.
ꢀ
<40 C. Elemental analyses were provided by Dr.
J. Theiner, Mikroanalytisches Laboratorium,
Institut fur Physikalische Chemie, University of
Vienna. Electrospray-ionisation mass spectra (ES-
MS) were obtained on a Finnigan TSQ7000
instrument in the positive ion mode using 1:1
MeOH±water in 0.1 mM NH4OAc containing 1%
HOAc as the matrix.
Allyl 2-acetamido-3,6-di-O-acetyl-2-deoxy-4-O-p-
methoxybenzyl-b-d-glucopyranoside (4).ÐA solu-
tion of 2 (200 mg, 0.52 mmol) and Ac2O (0.25 mL,
2.65 mmol) in dry pyridine (5 mL) was stirred for
24 h at room temperature. The reaction was quen-
ꢀ
Allyl 2-acetamido-2-deoxy-4-O-p-methoxybenzyl-
b-d-glucopyranoside (2) and allyl 2-acetamido-2-
deoxy-6-O-p-methoxybenzyl-b-d-glucopyranoside
(3).ÐA suspension of 1 (1.58 g, 4.15 mmol) and 4
A molecular sieves (0.5 g) in DMF (20 mL) was
stirred for 30 min under N2 at room temperature.
NaCNBH3 (5.2 g, 83 mmol) was added followed by
dropwise addition of CF3CO2H (3.23 mL,
42 mmol) in DMF (5 mL) for 30 min. The suspen-
sion was stirred for 24 h, then solid NaHCO3 (8 g)
was slowly added. The suspension was diluted with
CHCl3 (50 mL), ®ltered, and the ®ltrate was
washed with ice-cold water. The organic layer was
dried (Na2SO4), concentrated, and the residue was
puri®ed on a column of silica gel (8:1!4:1 EtOAc±
MeOH). Pooling of the fractions containing the
faster moving component and evaporation a€or-
ded 2. Yield: 568 mg (36%); colourless crystals, mp
ched by adding MeOH (2.5 mL) at 0 C. The solu-
tion was coevaporated with toluene (10 mL) and
taken to dryness. Crystallization of the residue
from EtOAc±hexane gave 4 as colourless crystals
ꢀ
(230 mg, 95%), mp 151±153 C (EtOAc±hexane);
[ꢁ]20
21ꢀ (c 0.6, CHCl3). H NMR (CDCl3): ꢂ
1
d
7.19±7.14 (m, 2 H, arom. H), 6.89±6.84 (m, 2 H,
arom. H), 5.84 (m, 1 H, CH=), 5.55 (d, 1 H, JNH,2
9.5 Hz, NH), 5.25 (dq, 1 H, =CH2trans), 5.17 (dq, 1
H, =CH2cis), 5.09 (dd, 1 H, J3,2 10.5, J3,4 8.5 Hz,
H-3), 4.56 and 4.47 (AB, 2 H, JA,B 10.5 Hz,
OCH2Ar), 4.42 (d, 1 H, J1,2 8.0 Hz, H-1), 4.35 (dd,
1 H, J6a,5 2.5, J6a,6b 12.0 Hz, H-6a), 4.31 (ddt, 1 H,
OCH2), 4.18 (dd, 1 H, J6b,5 4.5 Hz, H-6b), 4.08
(ddd, 1 H, H-2), 4.03 (ddt, 1 H, OCH2), 3.80 (s, 3
H, OCH3), 3.65 (t, 1 H, H-4), 3.53 (ddd, 1 H, J5,4
9.5 Hz, H-5), 2.06, 2.05, 1.95 (3 s, each 3 H, 3 Ac).
Anal. Calcd for C23H31NO9: C, 59.35; H, 6.71; N,
3.01. Found: C, 59.39; H, 6.86; N, 3.15.
ꢀ
227±229 C (CH2Cl2±hexane); [ꢁ]20
31ꢀ (c 1.2,
d
1
MeOH). H NMR (CD3OD): ꢂ 7.30±7.27 (m, 2 H,
arom. H), 6.90±6.86 (m, 2 H, arom. H), 5.89 (m, 1
H, CH=), 5.26 (dq, 1 H,=CH2trans), 5.13 (dq, 1
H,=CH2cis), 4.85 and 4.57 (AB, 2 H, JA,B 10.5 Hz,
OCH2Ar), 4.40 (d, 1 H, J1,2 8.5 Hz, H-1), 4.33 (ddt,
1H, OCH2), 4.05 (ddt, 1 H, OCH2), 3.81 (dd, 1 H,
J6a,5 2.0, J6a,6b 12.0 Hz, H-6a), 3.78 (s, 3 H, OCH3),
3.72 (dd, 1 H, J4,3 8.0, J4,5 10.0 Hz, H-4), 3.64 (dd,
1 H, J6b,5 5.0 Hz, H-6b), 3.61 (dd, 1 H, J2,1 8.5, J2,3
10.0 Hz, H-2), 3.38 (dd, 1 H, H-3), 3.26 (ddd, 1 H,
H-5), 1.98 (s, 3 H, Ac). Anal. Calcd for
C19H27NO7: C, 59.83; H, 7.13; N, 3.67. Found: C,
59.47; H, 7.09; N, 3.60.
Allyl 2-acetamido-3,6-di-O-acetyl-2-deoxy-b-d-
glucopyranoside (5).ÐA solution of 4 (300 mg,
0.64 mmol) in 18:1 CH2Cl2±H2O (7 mL) was trea-
ted with ceric ammonium nitrate (710 mg,
ꢀ
1.3 mmol) at 4 C with ultrasonication for 70 min.
NaHCO3 was added until the solution had pH 5.
The solution was dried (Na2SO4) and ®ltered.
Solids were washed thoroughly with EtOAc, the
organic phases were combined and concentrated.
Puri®cation of the residue on silica gel (5:5:1
CH2Cl2±hexane±EtOH) furnished 5 (159 mg, 72%)
as colourless crystals, mp 146±148 ꢀC (EtOAc±
hexane); [ꢁ]20
69ꢀ (c 0.5, CHCl3). 1H NMR
d
Further elution furnished 3 (652 mg, 41%) as
ꢀ
(CDCl3): ꢂ 5.87 (m, 1 H, CH=), 5.70 (d, 1 H, JNH,2
9.0 Hz, NH), 5.27 (dq, 1 H, =CH2trans), 5.19 (dq, 1
H, =CH2cis), 5.10 (dd, 1 H, J3,2 10.5, J3,4 8.5 Hz,
H-3), 4.57 (d, 1 H, J1,2 8.5 Hz, H-1), 4.51 (dd, 1 H,
J6a,5 4.0, J6a,6b 12.0 Hz, H-6a), 4.38 (ddt, 1 H,
OCH2), 4.31 (dd, 1 H, J6b,5 2.5 Hz, H-6b), 4.09
(ddt, 1 H, OCH2), 3.92 (ddd, 1 H, H-2), 3.62±3.54
(m, 1 H, H-4), 3.52 (ddd, 1 H, J5,4 10.0 Hz, H-5),
3.14 (d, 1 H, JOH,4 5.0 Hz, OH), 2.13, 2.11, and 1.96
colourless crystals, mp 130±132 C (CH2Cl2±hex-
ane); [ꢁ]20 +2ꢀ (c 1.1, MeOH). 1H NMR
d
(CD3OD): ꢂ 7.31±7.26 (m, 2 H, arom. H), 6.91±6.86
(m, 2 H, arom. H), 5.89 (m, 1 H, CH=), 5.29 (dq,
1 H, =CH2trans), 5.13 (dq, 1 H, =CH2cis), 4.52 (s,
2 H, OCH2Ar), 4.38 (d, 1 H, J1,2 8.5 Hz, H-1), 4.30
(ddt, 1 H, OCH2), 4.05 (ddt, 1 H, OCH2), 3.82 (dd,
1 H, J6a,5 2.0, J6a,6b 11.0 Hz, H-6a), 3.78 (s, 3 H,

J. Bartek et al./Carbohydrate Research 308 (1998) 259±273
265
ꢀ
(3 s, each 3 H, 3 Ac). Anal. Calcd for C15H23NO8:
C, 52.17; H, 6.71; N, 4.06. Found: C, 52.15; H,
6.61; N, 3.98.
(78%), colourless crystals, mp 91±92 C (hexane±
EtOAc); [ꢁ]20 +81ꢀ (c 0.7, CHCl3). H NMR
(CDCl3): ꢂ 8.83 (s, 1 H, =NH), 6.83 (d, 1 H, JNH,2
1
d
1,3,4,6-Tetra-O-acetyl-2-amino-2-deoxy-b-d-gal-
actopyranose hydrochloride (7).ÐA solution of 6
(3.5 g, 7.5 mmol) in acetone (30 mL) was heated to
re¯ux temperature. After addition of 1 M HCl
(11.3 mmol), the solution was cooled to 0 C. The
crystalline mass formed was ®ltered, washed with
9.0 Hz, NH), 6.52 (d, 1 H, J1,2 3.5 Hz, H-1), 5.54
(dd, 1 H, J4,3 3.0, J4,5 1.0 Hz, H-4), 5.42 (dd, 1 H,
J3,2 11.0 Hz, H-3), 4.72 (ddd, 1 H, H-2), 4.41 (dt, 1
H, J5,6a=J5,6b=6.5 Hz, H-5), 4.20 (dd, 1 H, J6a,6b
11.5 Hz, H-6a), 4.09 (dd, 1 H, H-6b), 2.22, 2.05,
and 2.03 (3 s, each 3 H, 3 Ac). Anal. Calcd for
C16H18Cl6N2O9: C, 32.30; H, 3.05; N, 4.70. Found:
C, 32.60; H, 3.06; N, 4.47.
Allyl (3,4,6-tri-O-acetyl-2-deoxy-2-trichloroaceta-
mido-b-d-galactopyranosyl)-(1!4)-2-acetamido-
3,6-di-O-acetyl-2-deoxy-b-d-glucopyranoside (10).Ð
A solution of TMSOTf (0.311 mmol) in toluene
(1 mL) was added at 0 ꢀC to a solution of 5 (72 mg,
0.208 mmol) and 9 (185 mg, 0.311 mmol) in dry
CH2Cl2 (5 mL) under N2. The solution was stirred
for 30 min at 0 ꢀC, then diluted with CH2Cl2
(50 mL) and washed with satd aq NaHCO3. The
organic layer was dried (Na2SO4), concentrated,
and the residue was applied on silica gel. Elution
with EtOAc gave 10 (60 mg, 37%) as the major
reaction product and 32 (not isolated)ꢀas side-pro-
duct. Colourless crystals, mp 211±213 C (hexane±
ꢀ
acetone, and dried to give 7 (2.9 g, 86%) as col-
ꢀ
ourless needles, mp 210 C (dec.); [ꢁ]20 +30ꢀ (c
d
1
0.3, H2O). H NMR (D2O): ꢂ 5.97 (d, 1 H, J1,2
9.0 Hz, H-1), 5.54 (dd, 1 H, J4,3 3.5, J4,5 1.0 Hz, H-
4), 5.34 (dd, 1 H, J3,2 11.0 Hz, H-3), 4.42 (t, 1 H,
J5,6a=J5,6b=5.5 Hz, H-5), 4.26±4.24 (m, 2 H, H-
6a,6b), 3.87 (dd, 1 H, H-2), 2.25, 2.22, 2.11, and
2.09 (4 s, each 3 H, 4 Ac). Anal. Calcd for
C14H22C1NO9: C, 43.81; H, 5.77; N, 3.65. Found:
C, 43.65; H, 5.51; N, 3.51.
1,3,4,6-Tetra-O-acetyl-2-deoxy-2-trichloroaceta-
mido-b-d-galactopyranose (8).ÐA suspension of
7 (200 mg, 0.52 mmol) in CH2Cl2 (5 mL) was
ꢀ
cooled to 0 C. Et3N (0.2 mL, 1.5 mmol) and tri-
chloroacetyl chloride (0.09 mL, 0.81 mmol) was
added. After 50 min the reaction mixture was dilu-
ted with CH2Cl2 (50 mL) and washed with satd aq
NaHCO3. The organic layer was dried (Na2SO4),
concentrated, and the residue was puri®ed on a
column of silica gel (2:1 toluene±EtOAc) to give 8
EtOAc); [ꢁ]20
38ꢀ (c 0.2, CHCl3). 1H NMR
d
(CDCl3): ꢂ 7.20 (d, 1 H, JNH ,2 9.5 Hz, NH0), 5.85
(m, 1 H, CH=), 5.72 (d, 1 H, JNH,2 9.5 Hz, NH),
0
0
5.37 (dd, 1 H, J4 ,3 3.4, J4 ,5 0.9 Hz, H-40), 5.26 (dq,
1 H, =CH2trans), 5.22 (dq, 1 H, =CH2cis), 5.20
0
0
0
0
(255 mg, 99%) as colourless syrup; [ꢁ]20 +4ꢀ (c
d
0.8, CHCl3). H NMR (CDCl3): ꢂ 6.84 (d, 1 H,
(dd, 1 H, J3 ,2 11.3 Hz, H-30), 5.12 (dd, 1 H, J3,2
1
0 0
0
0
JNH,2 9.5 Hz, NH), 5.84 (d, 1 H, J1,2 9.0 Hz, H-1),
5.42 (dd, 1 H, J4,3 3.3, J4,5 1.1 Hz, H-4), 5.25 (dd, 1
H, J3,2 11.3 Hz, H-3), 4.45 (ddd, 1 H, H-2), 4.20
(dd, 1 H, J6a,5 6.5, J6a,6b 11.5 Hz, H-6a), 4.14 (dd, 1
H, J6b,5 6.5 Hz, H-6b), 4.07 (dt, 1 H, H-5), 2.20,
2.14, 2.07, and 2.03 (4 s, each 3 H, 4 Ac). Anal.
Calcd for C16H20Cl3NO10: C, 39.00; H, 4.11; N,
2.85. Found: C, 39.67; H, 3.95; N, 2.72.
3,4,6-Tri-O-acetyl-2-deoxy-2-trichloroacetamido-
a-d-galactopyranosyl trichloroacetimidate (9).ÐA
solution of 8 (1.7 g, 3.45 mmol) in DMF (15 mL)
was stirred with hydrazine acetate (479 mg,
5.2 mmol) for 1 h at room temperature. The solu-
tion was diluted with CH2Cl2 (50 mL), washed with
aq NaHCO3, dried (Na2SO4), and evaporated to
dryness. A solution of the residue in CH2Cl2
(25 mL) was stirred with DBU (0.13 mL,
34.5 mmol) and trichloroacetonitrile (3.5 mL,
34.5 mmol) for 75 min. After concentration, the
residue was puri®ed on silica gel (2:1 hexane±
EtOAc containing 0.1% Et3N). Yield for 9: 1.6 g
9.5, J3,4 8.0 Hz, H-3), 4.64 (d, 1 H, J1 ,2 8.3 Hz, H-
10), 4.51 (d, 1 H, J1,2 7.4 Hz, H-1), 4.45 (dd, 1 H,
J6a,5 4.7, J6a,6b 12.1 Hz, H-6a), 4.35±4.28 (m, 2 H,
H-6b, OCH2), 4.18±4.00 (m, 5 H, H-2,20,6a0,6b0,
0
0
0
0
and OCH2), 3.91 (dt, 1 H, J5 ,6a =J5 ,6b =6.6 Hz,
H-50), 3.78 (t, 1 H, J4,5 8.0 Hz, H-4), 3.67±3.61 (m,
1 H, H-5), 2.17, 2.13, 2.08, 2.07, 1.99, and 1.987 (6
s, each 3 H, 6 Ac). Anal. Calcd for C29H39Cl3
N2O16: C, 44.77; H, 5.05; N, 3.60. Found: C, 45.17;
H, 4.92; N, 3.33.
Allyl (2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-
b-d-galactopyranosyl)-(1!4)-2-acetamido-3,6-di-
O-acetyl-2-deoxy-b-d-glucopyranoside (11).ÐA
solution of 10 (38 mg, 0.05 mmol) and NaBH4
(67 mg, 1.77 mmol) in EtOH (8 mL) was stirred for
ꢀ
48 h at 60 C. The solution was diluted with EtOH
ꢀ
(5 mL), cooled to 0 C and neutralized with HOAc
(0.1 mL). The solution was coevaporated ®ve times
with MeOH (10 mL portions) and taken to dry-
ness. The residue was dissolved in pyridine
(8 mL) and stirred with Ac2O (0.2 mL) for 60 h.

266
J. Bartek et al./Carbohydrate Research 308 (1998) 259±273
The solution was concentrated and the residue was
puri®ed on silica gel (10:1 EtOAc±MeOH). The
crude product obtained (28 mg) was further sub-
jected to LC on silica gel (10 mm, 10:1 EtOAc±
MeOH) which gave 11 as colourless crystals. Yield:
0.1 M methanolic NaOMe (1 mL) for 24 h at room
temperature. The solution was made neutral by
addition of Dowex 50 (H+) resin and ®ltered. The
®ltrate was evaporated and the residue was dis-
solved in water (2 mL) and lyophilized giving 13 as
22.5ꢀmg (68%), mp 223±225 ꢀC (EtOAc); [ꢁ]20
an ꢀamorphous solid. Yield: 11 mg (quant.); [ꢁ]20
d
d
23 (c 0.5, MeOH). 1H NMR (CDCl3): ꢂ 5.86 (m,
3 (c 0.6, H2O). H NMR (500 MHz, D2O): ꢂ
5.84 (m, 1 H, CH=), 5.24 (dq, 1 H, =CH2trans),
1
1 H, CH=), 5.78 (d, 1 H, JNH ,2 8.8 Hz, NH0), 5.77
0
0
0
(d, 1 H, JNH,2 9.8 Hz, NH), 5.32 (dd, 1 H, J4 ,3 3.5,
0
5.20 (dq, 1 H, =CH2cis), 4.50 (d, 1 H, J1,2 8.2 Hz,
0
0
H-1), 4.46 (d, 1 H, J1 ,2 8.4 Hz, H-10), 4.27 (ddt, 1
H, OCH2), 4.09 (ddt, 1 H, OCH2), 3.88 (dd, 1 H,
0
J4 ,5 <1.0 Hz, H-4 ), 5.26 (dq, 1 H, =CH2trans), 5.18
0
0
0
0
(dq, 1 H, =CH2cis), 5.15 (dd, 1 H, J3 ,2 11.5 Hz, H-
30), 5.12 (t, 1 H, J3,4=J3,2=8.5 Hz, H-3), 4.60 (d, 1
J4 ,3 3.3, J4 ,5 <1.0 Hz, H-40), 3.87 (dd, 1 H, J2 ,3
0
0
0
0
0
0
H, J1 ,2 8.3 Hz, H-10), 4.51 (d, 1 H, J1,2 7.3 Hz, H-
1), 4.41 (dd, 1 H, J6a,5 3.0, J6a,6b 12.0 Hz, H-6a),
4.35±4.28 (m, 1 H, OCH2), 4.31 (dd, 1 H, J6b,5
5.0 Hz, H-6b), 4.14±3.94 (m, 4 H, H-2,20,6b0, and
10.9 Hz, H-20), 3.79 (dd, 1 H, J6a,5 2.1, J6a,6b
12.1 Hz, H-6a), 3.74 (dd, 1 H, J2,3 11.8 Hz, H-2),
3.72±3.65 (m, 3 H, H-50,6a0,6b0), 3.70 (dd, 1 H, H-
30), 3.64 (dd, 1 H, J3,4 8.2 Hz, H-3), 3.60 (dd, 1 H,
J6b,5 5.6 Hz, H-6b), 3.56 (dd, 1 H, J4,5 9.6 Hz, H-4),
3.45 (ddd, 1 H, H-5), 2.01 and 1.97 (2 s, each 3 H, 2
Ac). ES-MS (C19H32N2O11): m/z 465.1 (M+H)+.
Allyl (2-chloroacetamido-2-deoxy-b-d-galacto-
pyranosyl)-(1!4)-2-acetamido-2-deoxy-b-d-gluco-
pyranoside (14).ÐA solution of 12 (3.5 mg,
4.9 mmol) in MeOH (2 mL) was stirred with 0.1 M
methanolic NaOMe (0.3 mL) for 24 h at room
temperature. Work-up as described for 13 gave 14
as amorphous powder. Yield 2.5 mg (ꢁ quant.);
0
0
0
0
0
0
OCH2), 4.09 (dd, 1 H, J6a ,5 6.5, J6a ,6b 11.0 Hz, H-
6a0), 3.87 (td, 1 H, J5 ,6b 6.5 Hz, H-50), 3.76 (t, 1 H,
J4,5 8.5 Hz, H-4), 3.69±3.64 (m, 1 H, H-5), 2.14,
2.13, 2.08, 2.05, 1.99, 1.96, and 1.958 (7 s, each 3
H, 7 Ac). Anal. Calcd for C29H42N2O16: C, 51.62;
H, 6.27; N, 4.15. Found: C, 51.51; H, 5.96; N, 4.00.
Allyl (3,4,6-tri-O-acetyl-2-chloroacetamido-2-
deoxy-b-d-galactopyranosyl)-(1!4)-2-acetamido-
3,6-di-O-acetyl-2-deoxy-b-d-glucopyranoside (12).Ð
A solution of 10 (7 mg, 9 mmol) in HOAc (2 mL)
was treated with zinc-dust (35 mg, 0.535 mmol) for
4 h at 110 ꢀC. The solution was diluted with
CH2Cl2 (50 mL), ®ltered, and the ®ltrate was con-
centrated. Puri®cation of the residue on silica gel
(EtOAc) gave 12 as an amorphous solid. Yield:
0
0
[ꢁ]20
4ꢀ (c 0.2, H2O). 1H NMR (D2O): ꢂ 5.90 (m,
d
1 H, CH=), 5.33±5.22 (m, 2 H, =CH2), 4.59 (d, 1
H, J1 ,2 8.3 Hz, H-10), 4.55 (d, 1 H, J1,2 8.1 Hz, H-
1), 4.32 (ddt, 1 H, OCH2), 4.21 and 4.18 (AB, 2 H,
JAB 14.3 Hz, ClCH2), 4.14 (ddt, 1 H, OCH2), 3.98
0
0
ꢀ
5 mg (78%). [ꢁ]20
23 (c 0.4, MeOH). H NMR
0
(dd, 1 H, J2 ,3 10.9 Hz, H-20), 3.95 (dd, 1 H, J4 ,3
1
0
0
0
0
d
(CDCl3): ꢂ 6.70 (d, 1 H, JNH ,2 8.5 Hz, NH0), 5.86
(m, 1 H, CH=), 5.72 (d, 1 H, JNH,2 9.0 Hz, NH),
2.8, J4 ,5 <1.0 Hz, H-40), 3.86 (dd, 1 H, J6a,5 2.6,
J6a,6b 11.9 Hz, H-6a), 3.82 (dd, 1 H, H-30), 3.79 (dd,
0
0
0
5.35 (dd, 1 H, J4 ,3 3.5, J4 ,5 1.0 Hz, H-40), 5.28 (dd,
1 H, J6a ,5 3.3, J6a ,6b 10.9 Hz, H-6a0), 3.77-3.61 (m,
6 H, H-2,3,4,6b,50,6b0), 3.50 (ddd, 1 H, H-5), 2.02
(s, 3 H, Ac). ES-MS (C19H31ClN2O11): m/z 499.1
(M+H)+.
0
0
0
0
0
0
0
0
1 H, J3 ,2 11.5 Hz, H-30), 5.26 (dq, 1 H,
=CH2trans), 5.19 (dq, 1 H, =CH2cis), 5.12 (dd, 1
0
0
0
0
H, J3,4 8.0, J3,2 9.5 Hz, H-3), 4.75 (d, 1 H, J1 ,2
7.2 Hz, H-10), 4.51 (d, 1 H, J1,2 7.2 Hz, H-1), 4.42
(dd, 1 H, J6a,5 3.0, J6a,6b 12.0 Hz, H-6a), 4.31 (ddt,
1 H, OCH2), 4.18 (dd, 1 H, J6b,5 5.6 Hz, H-6b),
4.14±4.00 (m, 4 H, H-2,6a0,6b0, and OCH2), 4.03 (s,
Allyl (2,3,4-tri-O-benzoyl-b-l-fucopyranosyl)-
(1!3)-2-acetamido-2-deoxy-4,6-O-p-methoxybenzyl-
idene-b-d-glucopyranoside (16).ÐCompound 15
(170 mg, 0.317 mmol) and 1 (105 mg, 0.264 mmol)
were dissolved in dry nitromethane (10 mL) under
N2. After addition of 4 A molecular sieves and
stirring for 0.5 h, Hg(CN)2 (72 mg, 0.285 mmol)
and HgBr2 (34 mg, 0.095 mmol) were added, and
stirring was continued for 14 h at ambient tem-
perature. The mixture was diluted with CH2Cl2
(50 mL) and ®ltered through Celite1. The ®ltrate
was washed with aq 10% KI and satd aq
NaHCO3 (2Â), dried over MgSO4, and con-
centrated. Puri®cation of the residue by column
2 H, ClCH2), 3.96 (dd, 1 H, H-20), 3.90 (dt, 1 H,
0
0
0
0
0
J5 ,6a =J5 ,6b =7.0 Hz, H-5 ), 3.78 (t, 1 H, J4,5
8.0 Hz, H-4), 3.63 (m, 1 H, H-5), 2.14, 2.12, 2.08,
2.06, 1.99, and 1.98 (6 s, each 3 H, 6 Ac). Anal.
Calcd for C29H41ClN2O16: C, 49.12; H, 5.83; N,
3.95. Found: C, 48.84; H, 5.77; N, 3.89.
Allyl (2-acetamido-2-deoxy-b-d-galactopyran-
osyl)-(1!4)-2-acetamido-2-deoxy-b-d-glucopyr-
anoside (13).ÐA solution of 11 (16 mg,
0.024 mmol) in MeOH (4 mL) was stirred with

J. Bartek et al./Carbohydrate Research 308 (1998) 259±273
267
chromatography (2:1 toluene±EtOAc) yielded 16
(175 mg, 75%) as crysꢀtals, mp 228±234 ꢀC (EtOAc±
Further elution gave 19 (1.49 g, 19%) as crystals,
158ꢀ (c
0.75, CHCl3). H NMR (CDCl3): ꢂ 7.89±6.88 (m,
ꢀ
mp 111±115 C (CH2Cl2±MeOH); [ꢁ]20
d
1
1
MeOH); [ꢁ]20
110 (c 0.8, CHCl3). H NMR
d
0
0
0
0
(CDCl3): ꢂ 8.09±6.96 (m, 19 H, arom. H), 5.92 (d, 1
23 H, arom. H), 5.85 (dd, 1 H, J3 ,2 11.0, J3 ,4
3.3 Hz, H-30), 5.58 (s, 1 H, OCHO), 5.53 (m, 1 H,
H, JNH,2 7.0 Hz, NH), 5.88 (m, 1 0H, CH=), 5.70
CH=), 5.49 (dd, 1 H, J4 ,5 1.0 Hz, H-40), 5.38 (dd,
0
0
0
0
0 0
(dd, 1 H, J3 ,4 3.5, J4 ,5 1.0 Hz, H-4 ), 5.67 (dd, 1 H,
J2 ,3 10.5, J2 ,1 8.0 Hz, H-20), 5.47 (dd, 1 H, H-30),
5.28 (dq, 1 H, =CH2trans), 5.21 (d, 1 H, J1,2 8.0 Hz,
H-1), 5.19 (dq, 1 H, =CH2cis), 5.07 (s, 1 H,
OCHO), 5.01 (d, 1 H, H-10), 4.64 (dd, 1 H, J3,2
10.0, J3,4 9.0 Hz, H-3), 4.32 (ddt, 1 H, OCH2), 4.24
(dd, 1 H, J6a,6b 10.0, J6b,5 4.5 Hz, H-6b), 4.10 (ddt,
1 H, J2 ,1 3.8 Hz, H-20), 5.16 (d, 1 H, H-10), 5.09 (d,
1 H, J1,2 8.5 Hz, H-1), 5.01 (dq, 1 H, =CH2trans),
4.96 (dd, 1 H, J3,2 10.5, J3,4 9.0 Hz, H-3), 4.92 (dq,
0
0
0
0
0
0
1 H, =CH2cis), 4.56 (dq, 1 H, J5 ,6 6.5 Hz, H-50),
4.40 (dd, 1 H, J6a,6b 10.0, J6b,5 4.5 Hz, H-6b), 4.33
(dd, 1 H, H-2), 4.20 (ddt, 1 H, OCH2), 3.91 (ddt, 1
H, OCH2), 3.87 (t, 1 H, J6a,5 ꢁ10 Hz, H-6a), 3.82
(m, 1 H, H-4), 3.78 (s, 3 H, OMe), 3.71 (dt, 1 H,
J5,4 ꢁ9.5 Hz, H-5), 0.55 (d, 3 H, H-60). Anal. Calcd
for C52H47NO15: C, 67.45; H, 5.12; N, 1.51.
Found: C, 67.44; H, 5.33; N, 1.52.
Allyl (3,4-O-isopropylidene-2-O-p-methoxy-
benzyl-a-l-fucopyranosyl)-(1!3)-2-acetamido-2-
deoxy-4,6-O-p-methoxybenzylidene-b-d-glucopyra-
noside (21).ÐCompound 1 (1.423 g, 3.75 mmol)
and 20 (1.656 g, 4.5 mmol) were dissolved in 5:1
CH2Cl2±DMF (96 mL) under N2. After addition
of 4 A molecular sieves, and stirring for 0.5 h,
IDCP (1.757 g) was added in 5 portions over
15 h, and stirring at room temperature was con-
tinued for further 25 h. Then the mixture was
diluted with CH2Cl2, and ®ltered through Celite1.
The ®ltrate was washed with aq 5% Na2S2O3,
dried over MgSO4, and concentrated. Puri®cation
of the residue by column chromatography (1:1
toluene±EtOAc) and crystallization from EtOAc±
0
0
1 H, OCH2), 4.07 (dq, 1 H, J5 ,6 6.5 Hz, H-50), 3.87
(s, 3 H, OMe), 3.60 (t, 1 H, J6a,5 10.0 Hz, H-6a),
3.49 (ddd, 1 H, J5,4 9.0 Hz, H-5), 3.37 (t, 1 H, H-4),
3.15 (ddd, 1 H, H-2), 1.99 (s, 3 H, Ac), 1.32 (d, 3
H, H-60). Anal. Calcd for C46H47NO14: C, 65.94;
H, 5.65; N, 1.67. Found: C, 65.61; H, 5.72; N,
1.67.
Allyl (2,3,4-tri-O-benzoyl-b-l-fucopyranosyl)-
(1!3)-2-deoxy-4,6-O-p-methoxybenzylidene-2-phthal-
imido-b-d-glucopyranoside (18) and allyl(2,3,4-tri-
O-benzoyl-a-l-fucopyranosyl)-(1!3)-2-deoxy-4,6-
O-p-methoxybenzylidene-2-phthalimido-b-d-gluco-
pyranoside (19).ÐDonor 15 (5.06 g, 9.38 mmol)
and compound 17 (3.95 g, 8.44 mmol) were dis-
solved in dry nitromethane (60 mL) under N2.
After addition of 4 A molecular sieves and stirring
for 0.5 h, Hg(CN)2 (2.13 g, 8.44 mmol) and HgBr2
(1.01 g, 2.81 mmol) were added, and stirring at
ambient temperature was continued for 24 h. The
mixture was diluted with CH2Cl2 and ®ltered
through Celite1. The ®ltrate was washed with aq
10% KI and satd aq NaHCO3 (2Â), dried over
MgSO4, and concentrated. Puri®cation of the resi-
due by column chromatography (12:1 toluene±
EtOAc) yielded 18 (1.46 g, 19%) as crystals, mp
0
0
ꢀ
hexane yieꢀlded 21 (1.262 g, 49%), mp 184±186 C;
1
[ꢁ]20
78 (c 1.1, CHCl3). H NMR (CDCl3): ꢂ
d
7.43±7.38 (m, 2 H, arom. H), 7.30±7.25 (m, 2 H,
arom. H), 6.91±6.85 (m, 4 H, arom. H), 5.84 (m,
1 H, CH=), 5.75 (d, 1 H, JNH,2 7.0 Hz, NH),
5.46 (s, 1 H, OCHO), 5.25 (dq, 1 H, =CH2trans),
5.17 (dq, 1 H, =CH2cis), 5.07 (d, 1 H, J1,2
8.5 Hz, H-1), 4.90 (d, 1 H, J1 ,2 3.5 Hz, H-10), 4.84
(d, 1 H, Jgem 11.5 Hz, OCH2Ar), 4.35 (dq, 1 H,
J5 ,6 6.5, J5 ,4 2.5 Hz, H-50), 4.35 (d, 1 H,
OCH2Ar), 4.23 (dd, 1 H, J3 ,2 8.0, J3 ,4 5.5 Hz,
H-30), 4.40±4.22 (m, 3 H, H-3,6a, OCH2), 4.06
(ddt, 1 H, OCH2), 3.93 (dd, 1 H, H-40), 3.80 (2 s,
each 3 H, 2 OMe), 3.74 (br t, 1 H, J6a,6b=J6b,5
ꢁ10 Hz, H-6b), 3.53 (dd, 1 H, H-20), 3.45±3.56 (m,
2 H, H-4, 5), 3.23 (ddd, 1 H, J2,3 10.0 Hz, H-2),
1.73 (s, 3 H, Ac), 1.43, and 1.31 (2 s, each 3 H,
Me2C), 0.69 (d, 3 H, H-60). Anal. Calcd for
C36H47NO12: C, 63.05; H, 6.91; N, 2.04. Found: C,
63.07; H, 6.92; N, 2.02.
ꢀ
1
131±135 C (CH2Cl2±MeOH); [ꢁ]20
CHCl3). H NMR (CDCl3): ꢂ 7.82±6.93 (m, 23 H,
0
arom. H), 5.72 (m, 1 H, CH=), 5.49 (dd, 1 H, J2 ,3
10.3, J2 ,1 7.8 Hz, H-20), 5.42 (dd, 1 H, J4 ,3 3.5,
38ꢀ (c 0.57,
d
0
0
0
0
0
0
0
J4 ,5 <1.0 Hz, H-40), 5.42 (d, 1 H, J1,2 8.5 Hz, H-1),
5.32 (dd, 1 H, H-30), 5.20 (s, 1 H, OCHO), 5.15 (dq,
1 H, =CH2trans), 5.05 (dq, 1 H, =CH2cis), 4.89 (d,
1 H, H-10), 4.71 (dd, 1 H, J3,2 10.0, J3,4 9.0 Hz, H-
3), 4.33 (dd, 1 H, H-2), 4.31 (dd, 1 H, J6a,6b 10.0,
J6b,5 4.5 Hz, H-6b), 4.28 (ddt, 1 H, OCH2), 4.04
(ddt, 1 H, OCH2), 3.88 (s, 3 H, OMe), 3.82 (dq, 1
0
0
0
0
0
0
0
0
0
0
H, J5 ,6 6.5 Hz, H-50), 3.72 (t, 1 H, J6a,5 ꢁ10 Hz, H-
6a), 3.65±3.55 (m, 2 H, H-4,5), 0.55 (d, 3 H, H-60).
Anal. Calcd for C52H47NO15: C, 67.45; H, 5.12; N,
1.51. Found: C, 67.03; H, 5.28; N, 1.48.
0
0

268
J. Bartek et al./Carbohydrate Research 308 (1998) 259±273
Allyl (3,4-O-isopropylidene-2-O-p-methoxybenzyl-
OCH2), 3.61 (ddd, 1 H, H-5), 3.25 (m, 1 H, H-2),
2.15, 2.11, 2.09, 2.00, 1.99, and 1.98 (6 s, each 3 H,
6 Ac), 1.07 (d, 3 H, H-60). Anal. Calcd for
C27H39NO15: C, 52.51; H, 6.36; N, 2.27. Found: C,
52.44; H, 6.38; N, 2.10.
a-l-fucopyranosyl)-(1!3)-2-acetamido-2-deoxy-b-
d-glucopyranoside (22).ÐTo a solution of 21
(590 mg, 0.86 mmol) in 9:1 CH3CN±water (150 mL)
was added ceric ammonium nitrate (456 mg,
0.833 mmol), and the mixture was stirred for 1 h at
room temperature. Then, NaHCO3 (0.5 g) was
added, and the mixture was concentrated to dry-
ness. The residue was puri®ed by column chroma-
tography (9:1 EtOAc±MeOH) to give 22 (267 mg,
55%), which crystallized from Et2O±CH2Cl2, mp
Allyl (a-l-fucopyranosyl)-(1!3)-2-acetamido-2-
deoxy-b-d-glucopyranoside (24).ÐA solution of 23
(7.2 mg) in dry MeOH (5 mL) was stirred with
0.1 M methanolic NaOMe (0.5 mL) for 2 h at room
temperature. The pH of the solution was adjusted
to 7.0 by addition of Dowex 50 (H+) resin, the
resin was ®ltered o€ and the ®ltrate was taken to
dryness. Puri®cation of the residue on a Bio-Gel P-
2 column (1.6Â80 cm, aq 5% EtOH) gave 24 as
ꢀ
ꢀ
20
1
175±177 C; [ꢁ]d
45 (c 0.6, CHCl3). H NMR
(CDCl3): ꢂ 7.31±7.25 (m, 2 H, arom. H), 6.92±6.86
(m, 2 H, arom. H), 5.87 (m, 1 H, CH=), 5.56 (d, 1
H, JNH,2 7.5 Hz, NH), 5.27 (dq, 1 H, =CH2trans),
5.19 (dq, 1 H, =CH2cis), 4.94 (d, 1 H, J1,2 8.5 Hz,
amorphous solid. Yield: 4.1 mg (87%); [ꢁ]20
105 (c 0.4, H2O). H NMR (D2O): ꢂ 5.92 (m, 1
d
ꢀ
1
H-1), 4.85 (d, 1 H, J1 ,2 3.5 Hz, H-10), 4.72 (d, 1 H,
J 11.7 Hz, OCH2Ar), 4.60 (d, 1 H, OCH2Ar), 4.35
H, CH=), 5.32 (dq, 1 H, =CH2trans), 5.28 (dq, 1
0
0
H, =CH2cis), 5.00 (d, 1 H, J1 ,2 4.1 Hz, H-10), 4.59
(d, 1 H, J1,2 8.6 Hz, H-1), 4.37 (m, 1 H, OCH2),
4.35 (m, 1 H, H-50), 4.18 (m, 1 H, OCH2), 3.96 (dd,
1 H, J6a,5 2.2, J6a,6b 12.4 Hz, H-6a), 3.87 (dd, 1 H,
0
0
(dq, 1 H, J5 ,6 6.5, J5 ,4 2.0 Hz, H-50), 4.32 (m, 1 H,
0
0
0
0
H-30), 4.31 (ddt, 1 H, OCH2), 4.10 (ddt, 1 H,
OCH2), 4.06 (dd, 1 H, J4 ,3 6.5 Hz, H-40), 3.96 (dd,
1 H, J3,2 10.5, J3,4 8.0 Hz, H-3), 3.90 (dd, 1 H, J6a,6b
ꢁ 12.0 Hz, H-6b), 3.81 (s, 3 H, OMe), 3.80 (dd, 1
0
0
H-2), 3.85 (dd, 1 H, J3 ,2 10.1, J3 ,4 3.2 Hz, H-30),
0
0
0
0
3.8 (dd, 1 H, J4 ,5 1.0 Hz, H-40), 3.77 (dd, 1 H, J6b,5
5.4 Hz, H-6b), 3.71 (dd, 1 H, H-20), 3.66 (dd, 1 H,
J3,2 10.1, J3,4 9.8 Hz, H-3), 3.54 (t, 1 H, J4,5 9.8 Hz,
H-4), 3.50 (m, 1 H, H-5), 2.04 (s, 3 H, Ac), 1.15 (d,
0
0
H, H-6a), 3.58 (dd, 1 H, J2 ,3 3.5 Hz, H-20), 3.48
(m, 1 H, J4,5 9.0 Hz, H-4), 3.40 (ddd, 1 H, J5,6a 5.0,
J5,6b 3.0 Hz, H-5), 3.26 (ddd, 1 H, H-2), 2.20 (br m,
2 H, OH), 1.77 (s, 3 H, Ac), 1.43 and 1.34 (2 s, each
3 H, Me2C), 1.31 (d, 3 H, H-60). Anal. Calcd for
C28H41NO11: C, 59.25; H, 7.28; N, 2.47. Found: C,
59.26; H, 7.12; N, 2.40.
0
0
3 H, J6 ,5 6.6 Hz, H-60).
0
0
Allyl (3,4-O-isopropylidene-2-O-p-methoxybenzyl-
a-l-fucopyranosyl)-(1!3)-2-acetamido-2-deoxy-6-
O-p-methoxybenzyl-b-d-glucopyranoside (25), allyl
(3,4-O-isopropylidene-2-O-p-methoxybenzyl-b-l-
fucopyranosyl)-(1!3)-2-acetamido-2-deoxy-6-O-p-
methoxybenzyl-b-d-glucopyranoside (26) and allyl
(3,4-O-isopropylidene -2-O-p-methoxybenzyl-a-l-
fucopyranosyl)-(1!4)-2-acetamido-2-deoxy-6-O-p-
methoxybenzyl-b-d-glucopyranoside (28).ÐA sus-
pension of 3 (600 mg, 1.57 mmol), 20 (636 mg,
1.73 mmol) and 4 A molecular sieves (1 g) in 1:5
dry DMF±CH2Cl2 (14 mL) was stirred at room
temperature for 75 min under N2. Iodonium di-
sym-collidinium perchlorate (IDCP, 785 mg,
1.57 mmol) was added in 7 portions over a period
of 7 h. Stirring was continued for one additional
hour, the suspension was diluted with CH2Cl2
(50 mL) and ®ltered through Celite1. The ®ltrate
was washed twice with aq 5% Na2S2O3, dried
(Na2SO4), and concentrated. Puri®cation of the
residue on silica gel (1:3 toluene±EtOAc) gave 25 as
colourless crystals. Yield: 260 mg (32%), mp 162±
Allyl (2,3,4-tri-O-acetyl-a-l-fucopyranosyl)-(1!3)-
2-acetamido-3,6-di-O-acetyl-2-deoxy-b-d-glucopy-
ranoside (23).ÐA solution of 21 (15 mg,
0.021 mmol) in 18:1 CH2Cl2±water (9 mL) was vig-
orously stirred with DDQ (21 mg, 0.093 mmol) at
room temperature for 24 h. The solution was taken
to dryness, the residue was dissolved in pyridine
(6 mL) and stirred with Ac2O (1.8 mL) for 20 h at
room temperature. MeOH (1 mL) was added and
the solution was concentrated. Puri®cation of the
residue on silica gel (EtOAc) gave 23 as colourless
57ꢀ (c 0.7,
20
syrup. Yield: 12 mg (92%); [ꢁ]d
CHCl3). 1H NMR (CDCl3): ꢂ 5.87 (m, 1 H, CH=),
0
5.70 (d, 1 H, JNH,2 7.1 Hz, NH), 5.31 (dd, 1 H, J3 ,4
0
3.2, J3 ,2 11.0 Hz, H-30), 5.27 (dq, 1 H, =CH2trans),
0
0
5.26 (d, 1 H, H-10), 5.24 (dd, 1 H, J5 ,4 1.5 Hz, H-
0
0
40), 5.21 (dq, 1 H, =CH2cis), 5.09 (dd, 1 H, J1 ,2
0
0
3.4 Hz, H-20), 5.03 (d, 1 H, J1,2 8.0 Hz, H-1), 4.96
(t, 1 H, J3,4 ꢁ J4,5=9.3 Hz, H-4), 4.45 (t, 1 H, J3,2
9.3 Hz, H-3), 4.32 (m, 1 H, OCH2), 4.19 (dd, 1 H,
J6a,6b 12.5, J6a,5 5.5 Hz, H-6a), 4.16 (m, 1 H, H-50),
4.08 (dd, 1 H, J6b,5 2.9 Hz, H-6b), 4.08 (m, 1 H,
164 ꢀC (EtOAc±hexane); [ꢁ]20
CHCl3). H NMR (CDCl3): ꢂ 7.29±7.25 (m, 4 H,
arom. H), 6.89±6.85 (m, 4 H, arom. H), 5.88 (m, 1
59ꢀ (c 0.5,
d
1

J. Bartek et al./Carbohydrate Research 308 (1998) 259±273
269
H, CH=), 5.51 (d, 1 H, JNH,2 7.7 Hz, NH), 5.25
(dq, 1 H, =CH2trans), 5.18±5.14 (m, 1 H,
=CH2cis), 4.89 (d, 1 H, J1,2 8.2 Hz, H-1), 4.86 (d, 1
each 3 H, 2 OMe), 3.74 (dd, 1 H, J6a,5 1.9, J6a,6b
10.5 Hz, H-6a), 3.69 (dd, 1 H, J6b,5 3.9 Hz, H-6b),
3.57±3.47 (m, 4 H, H-4,5,20, and OH), 3.31 (ddd, 1
H, J2,3 10.0 Hz, H-2), 2.01 (s, 3 H, Ac), 1.42 and
1.34 (2 s, each 3 H, Me2C), 1.29 (d, 3 H, H-60).
Anal. Calcd for C36H49NO12: C, 62.87; H, 7.18; N,
2.04. Found: C, 63.08; H, 7.34; N, 1.99. Further
elution a€orded unreacted 3 (150 mg, 25%).
Acetylation of 25.ÐA solution of 25 (5 mg) in
pyridine (1 mL) was treated with Ac2O (0.1 mL) for
64 h at room temperature. MeOH (0.2 mL) was
added and the solution was taken to dryness. Pur-
i®cation of the residue on silica gel (1:2 toluene±
EtOAc) furnished 27 as syrup. Yield: 5 mg (95%).
1H NMR (CDCl3): ꢂ 7.27±7.23 (m, 4 H, arom. H),
6.91±6.85 (m, 4 H, arom. H), 5.90±5.78 (m, 1 H,
CH=), 5.81 (d, 1 H, JNH,2 7.5 Hz, NH), 5.24 (dq, 1
H, =CH2trans), 5.15 (dq, 1 H, =CH2cis), 4.88 (t, 1
H, J4,3=J4,5=9.0 Hz, H-4), 4.78 and 4.58 (AB, 2
H, J1 ,2 3.5 Hz, H-10), 4.72 and 4.60 (AB, 2 H, JAB
0
0
12.0 Hz, OCH2Ar), 4.59±4.51 (m, 2 H,0 OCH2Ar),
0
0
0
0
4.36 (dq, 1 H, J5 ,4 2.5, J5 ,6 6.7 Hz, H-5 ), 4.31 (dd,
0
0
0
0
0
1 H, J3 ,2 6.7, J3 ,4 6.0 Hz, H-3 ), 4.39±4.29 (m, 1 H,
OCH2), 4.11±4.04 (m, 1 H, OCH2), 4.05 (dd, 1 H,
H-40), 3.96 (br dd, 1 H, H-3), 3.83±3.80 (m, 8 H, 2
OMe, H-6a,6b), 3.68 (ddd, 1 H J5,6a 5.0, J5,4
10.4 Hz, H-5), 3.75 (dd, 1 H, H-20), 3.48±3.43 (m, 2
H, H-4, OH), 3.26 (m, 1 H, H-2), 1.76 (s, 3 H, Ac),
1.42 and 1.33 (2 s, each 3 H, Me2C), 1.31 (d, 3 H,
H-60). Anal. Calcd for C36H49NO12: C, 62.87; H,
7.18; N, 2.04. Found: C, 62.94; H, 7.12; N, 1.99.
The column was then eluted with 5:1 EtOAc±
MeOH and the eluate was concentrated. The resi-
due was puri®ed on silica gel (10:10:1 CH2Cl2±
hexane±EtOH), which a€orded 26 as colourless
crystals. Yield: 33 mg (4%); mp 171±173 ꢀC
(CH2Cl2±hexane); [ꢁ]20
0
H, JAB 11.5 Hz, OCH2Ar), 4.76 (d, 1 H, J1 ,2
0
8ꢀ (c 0.6, CHCl3). H
3.0 Hz, H-1 ), 4.64 (d, 1 H, J1,2 8.3 Hz, H-1), 4.44
and 4.47 (AB, 2 H, JAB 11.0 Hz, OCH2Ar), 4.32
1
d
NMR (CDCl3): ꢂ 7.26 (m, 4 H, arom. H), 6.84 (m,
4 H, arom. H), 5.91 (m, 1 H, CH=), 5.78 (d, 1 H,
JNH,2 6.8 Hz, NH), 5.27 (dq, 1 H, =CH2trans), 5.17
(dq, 1 H, =CH2cis), 5.04 (d, 1 H, J1,2 8.1 Hz, H-1),
4.85 and 4.66 (AB, 2 H, JAB 11.5 Hz, OCH2Ar),
0
0
0
0
(ddt, 1 H, OCH2), 4.20 (dd, 1 H, J3 ,2 8.0, J3 ,4
5.5 Hz, H-30), 4.15 (dq, 1 H, J5 ,6 6.7 Hz, H-50),
4.07 (ddt, 1 H, OCH2), 3.97 (dd, 1 H, H-40), 3.81
and 3.80 (2 s, each 3 H, 2 OMe), 3.82±3.76 (m, 1 H,
H-3), 3.56±3.51 (m, 4 H, H-2,5,6a,6b), 3.47 (dd, 1
H, H-20), 1.98 (s, 6 H, 2 Ac), 1.44 and 1.33 (2 s,
each 3 H, Me2C), 1.23 (d, 3 H, H-60).
0
0
0
0
4.52 (s, 2 H, OCH2Ar), 4.41 (d, 1 H, J1 ,2 8.3 Hz,
H-10), 4.34 (ddt, 1 H, OCH2), 4.23 (br dd, 1 H, H-
3), 4.18 (dd, 1 H, J3 ,2 7.0, J3 ,4 5.5 Hz, H-30), 4.09
0
0
0
0
0
(ddt, 1 H, OCH2), 3.97 (dd, 1 H, J4 ,5 2.1 Hz, H-
0
Acetylation of 28.ÐCompound 28 (5 mg) was
treated as described for 27 to give 5 mg (95%) of 29
as a syrup. 1H NMR (CDCl3): ꢂ 7.25±7.18 (m, 4 H,
arom. H), 6.87±6.82 (m, 4 H arom. H), 5.86 (m, 1
H, CH=), 5.45 (d, 1 H, JNH,2 9.0 Hz, NH), 5.26
(dq, 1 H, =CH2trans), 5.17 (dq, 1 H, =CH2cis),
5.03 (dd, 1 H, J3,2 10.5, J3,4 8.5 Hz, H-3), 4.81 (d, 1
40), 3.86±3.78 (m, 1 H, H-6a), 3.82 (dq, 1 H, J5 ,6
0
0
6.6 Hz, H-50), 3.79 and 3.76 (2 s, each 3 H, 2 OMe),
3.73 (dd, 1 H, J6b,5 2.0, J6b,6a 11.0 Hz, H-6b), 3.62±
3.56 (m, 1 H, H-5), 3.52±3.47 (m, 2 H, H-4, OH),
3.43 (dd, 1 H, H-20), 3.04 (br td, 1 H, H-2), 1.97 (s,
3 H, Ac), 1.44 and 1.34 (2 s, each 3 H, Me2C), 1.36
(d, 3 H, H-60). Anal. Calcd for C36H49NO12: C,
62.87; H, 7.18; N, 2.04. Found: C, 62.89; H, 7.19;
N, 2.00.
H, J1 ,2 3.5 Hz, H-10), 4.67 and 4.53 (AB, 2 H, JAB
11.5 Hz, OCH2Ar), 4.47 (d, 1 H, J1,2 8.0 Hz, H-1),
4.41 and 4.35 (AB, 2 H, JAB 11.7 Hz, OCH2Ar),
0
0
0
0
0
0
Further elution gave 28 as crystals. Yield: 90 mg
72ꢀ (c
0.6, CHCl3). H NMR (CDCl3): ꢂ 7.22±7.19 (m, 4
H, arom. H), 6.86±6.81 (m, 4 H, arom. H), 5.90 (m,
1 H, CH=), 5.73 (d, 1 H, JNH,2 6.7 Hz, NH), 5.28
(dq, 1 H, =CH2trans), 5.19 (dq, 1 H, =CH2cis),
4.32 (ddt, 1 H, OCH2), 4.15 (dd, 1 H, J3 ,2 8.0, J3 ,4
ꢀ
(11%); mp 160 C (EtOAc±hexane); [ꢁ]20
5.5 Hz, H-30), 4.11±4.03 (m, 2 H, H-50, OCH2),
4.01±3.90 (m, 3 H, H-2,6a,40), 3.79 and 3.78 (2 s,
each 3 H, 2 OMe), 3.67 (dd, 1 H, J6b,5 5.5, J6b,6a
9.5 Hz, H-6b), 3.63 (dd, 1 H, J4,5 9.5 Hz, H-4), 3.52
(ddd, 1 H, J5,6a 2.0 Hz, H-5), 3.42 (dd, 1 H, H-20),
2.06 and 1.92 (2 s, each 3 H, 2 Ac), 1.42 and 1.33 (2
s, each 3 H, Me2C), 1.23 (d, 3 H, H-60).
Allyl (2,3,4-tri-O-acetyl-a-l-fucopyranosyl)-(1!4)-
2-acetamido-3,6-di-O-acetyl-2-deoxy-b-d-glucopyr-
anoside (30).ÐA solution of 28 (28 mg,
0.041 mmol) in CH2Cl2 (7 mL) was cooled to
20 ꢀC and treated with aq 90% CF3CO2H
d
1
4.88 (d, 1 H, J1 ,2 3.5 Hz, H-10), 4.87 (d, 1 H, J1,2
7.6 Hz, H-1), 4.61 and 4.55 (AB, 2 H, JAB 11.0 Hz,
OCH2Ar), 4.43 and 4.38 (AB, 2 H, JAB 11.0 Hz,
0
0
0
0
0
0
OCH2Ar), 4.39 (dq, 1 H, J5 ,4 2.5, J5 ,6 6.7 Hz, H-
50), 4.40±4.31 (m, 1 H, OCH2), 4.26 (t, 1 H,
J3 ,4 =J3 ,2 =5.5 Hz, H-3 ), 4.11±4.01 (m, 2 H, H-3,
OCH2), 4.02 (dd, 1 H, H-40), 3.79 and 3.77 (2 s,
0
0
0
0
0

270
J. Bartek et al./Carbohydrate Research 308 (1998) 259±273
(0.175 mL) for 2.5 h. After concentration, the resi-
due was dissolved in pyridine (6 mL) and treated
with Ac2O ꢀ(3 mL) for 96 h. MeOH (5 mL) was
added at 0 C and the solution was concentrated.
Puri®cation of the residue on silica gel (EtOAc)
by ®ltration, and the ®ltrate was washed with satd
aq NaHCO3, dried (Na2SO4), and concentrated.
Puri®cation of the residue on silica gel (1:2 toluene±
EtOAc) gave 2-trichloromethyl-4,5-dihydro-(3,4,
6-tri-O-acetyl-1,2-dideoxy-ꢁ-d-galactopyranoso)-
[2,1-d]-1,3-oxazole (32) as a syrup. Yield: 65 mg
(36%); [ꢁ]20d+108ꢀ (c 0.6, CHCl3). 1H NMR
(CDCl3): ꢂ 6.33 (d, 1 H, J1,2 7.0 Hz, H-1), 5.48 (dd, 1
H, J4,3 3.7, J4,5 2.5 Hz, H-4), 4.99 (dd, 1 H, J3,2
7.5 Hz, H-3), 4.31 (td, 1 H, J5,6a=J5,6b=6.5 Hz, H-
5), 4.29 (dd, 1 H, H-2), 4.22 (dd, 1 H, J6a,6b 11.5 Hz,
H-6a), 4.15 (dd, 1 H, H-6b), 2.15, 2.09, and 2.08 (3 s,
each 3 H, 3 Ac).
furnꢀished 30 as a syrup. Yield: 21 mg (83%); [ꢁ]20
d
93 (c 0.8, CHCl3). 1H NMR (CDCl3): ꢂ 5.86 (m,
1 H, CH=), 5.46 (d, 1 H, JNH,2 9.0 Hz, NH), 5.31±
5.10 (m, 6 H, H-10,20,30,40, and =CH2), 5.08 (dd, 1
H, J3,2 10.5, J3,4 8.0 Hz, H-3), 4.66 (dd, 1 H, J6a,5
2.0, J6a,6b 11.5 Hz, H-6a), 4.55 (d, 1 H, J1,2 8.5 Hz,
H-1), 4.32 (ddt, 1 H, OCH2), 4.13 (dd, 1 H, J6b,5
0
0
0
0
5.5 Hz, H-6b), 4.10 (dd, 1 H, J5 ,4 1.5, J5 ,6 6.5 Hz,
H-50), 4.08 (ddt, 1 H, OCH2), 3.92 (ddd, 1 H, H-2),
3.71±3.63 (m, 1 H, H-5), 3.60 (dd, 1 H, J4,5 9.5 Hz,
H-4), 2.15, 2.11, 2.09, 2.07, 1.98, and 1.94 (6 s, each
3 H, 6 Ac), 1.06 (d, 3 H, H-60). Anal. Calcd for
C27H39NO15: C, 52.51; H, 6.36; N, 2.27. Found: C,
52.64; H, 6.28; N, 2.18.
Further elution a€orded 33 as a syrup. Yield:
67ꢀ (c 0.6, CHCl3). 1H NMR
90 mg (41%); [ꢁ]20
d
(CDCl3): ꢂ 7.31±7.25 (m, 4 H, arom. H), 6.94±6.91
(m, 2 H, arom. H), 6.88±6.85 (m, 2 H, arom. H),
6.46 (d, 1 H, JNH ,2 9.0 Hz, NH00), 6.08 (d, 1 H,
JNH,2 8.0 Hz, NH), 5.83 (m, 1 H, CH=), 5.22 (dd,
00 00
1 H, J4 ,3 3.5, J4 ,5 1.0 Hz, H-400), 5.23 (dq, 1 H,
00 00
00 00
Allyl (a-l-fucopyranosyl)-(1!4)-2-acetamido-2-
deoxy-b-d-glucopyranoside (31).ÐA solution of 30
(9.8 mg) in dry MeOH (6 mL) was stirred with
0.1 M methanolic NaOMe (0.8 mL) for 24 h at
room temperature. The pH of the solution was
adjusted to 7.0 by addition of Dowex 50 (H+)
resin, the resin was ®ltered o€, and the ®ltrate was
taken to dryness to give 31 as an amorphous solid.
=CH2trans), 5.13 (dq, 1 H, =CH2cis), 5.06 (d, 1 H,
J1 ,2 3.4 Hz, H-10), 4.98 (dd, 1 H, J3 ,2 11.3 Hz, H-
300), 4.78±4.73 (m, 2 H, H-1, OCH2Ar), 4.63 (d, 1
H, JAB 12.0 Hz, OCH2Ar), 4.63 and 4.36 (AB, 2 H,
0
0
00 00
00 00
JAB 12.0 Hz, OCH2Ar), 4.59 (d, 1 H, J1 ,2 9.0 Hz,
H-100), 4.53 (dq, 1 H, J5 ,4 2.5, J5 ,6 6.7 Hz, H-50),
0
0
0
0
4.25 (dd, 1 H, J3 ,2 7.5, J3 ,4 6.0 Hz, H-30), 4.20 (dd,
0
0
0
0
Yield: 6.5 mg (ꢁ quant.); [ꢁ]20
99ꢀ (c 0.7, H2O).
1 H, J6a ,5 8.0, J6a ,6b 11.0 Hz, H-6a00), 4.28±4.17
(m, 1 H, OCH2), 4.10±3.96 (m, 6 H, H-
3,4,40,200,6b00, and OCH2), 3.81 and 3.80 (2 s, each 3
H, 2 OMe), 3.74±3.60 (m, 4 H, H-2,6a,6b,500), 3.57
(dd, 1 H, H-20), 3.54±3.49 (m, 1 H, H-5), 2.15, 2.06,
1.99, and 1.90 (4 s, each 3 H, 4 Ac), 1.43 and 1.35
(2 s, each 3 H, Me2C), 1.35 (d, 3 H, H-60). Anal.
Calcd for C50H65Cl3N2O20: C, 53.59; H, 5.85; N,
2.50. Found: C, 53.57; H, 6.02; N, 2.33. Further
elution of the column gave unreacted starting
material 25 (47 mg, 26%).
00 00
00
00
d
1H NMR (D2O): ꢂ 5.91 (m, 1 H, CH=), 5.34±5.23
(m, 2 H, =CH2), 4.95 (d, 1 H, J1 ,2 3.5 Hz, H-10),
4.56 (d, 1 H, J1,2 8.0 Hz, H-1), 4.33 (ddt, 1 H,
0
0
OCH2), 4.34 (dq, 1 H, J5 ,4 1.5, J5 ,6 6.5 Hz, H-50),
4.15 (ddt, 1 H, OCH2), 3.98 (dd, 1 H, J6a,5 1.8,
J6a,6b 12.5 Hz, H-6a), 3.87±3.80 (m, 2 H, H-6b,20),
0
0
0
0
3.80 (dd, 1 H, J4 ,3 3.5 Hz, H-40), 3.72 (dd, 1 H, J2,3
10.0 Hz, H-2), 3.77±3.50 (m, 4 H, H-3,4,5,30),
2.03 (s, 3 H, Ac), 1.15 (d, 3 H, H-60). ES-MS
(C17H29NO10): m/z 407.9 (M+H)+.
0
0
Allyl (3,4-O-isopropylidene-2-O-p-methoxy-
benzyl-a-l-fucopyranosyl)-(1!3)-[(3,4,6-tri-O-acetyl-
2-deoxy-2-trichloroacetamido-b-d-galactopyrano-
syl-(1!4)]-2-acetamido-2-deoxy-6-O-p-methoxy-
benzyl-b-d-glucopyranoside (33).ÐA suspension of
25 (183 mg, 0.266 mmol), 9 (240 mg, 0.403 mmol),
N,N,N0,N0-tetramethyl urea (0.035 ml, 0.292 mmol)
and 4 A molecular sieves (500 mg) in CH2Cl2
(8 mL) containing toluene (0.4 mL) was stirred for
45 min at room temperature. Then Sn(OTf)2
(146 mg, 0.348 mmol) was added in two portions
during 30 min, and the suspension was stirred
for one additional hour. Et3N (0.075 mL) and
CH2Cl2 (50 mL) were added, solids were removed
Allyl (2-O-acetyl-3,4-O-isopropylidene-a-l-fuco-
pyranosyl)-(1!3)-[(3,4,6-tri-O-acetyl-2-deoxy-2-
trichloroacetamido-b-d-galactopyranosyl)-(1!4)]-
2-acetamido-6-O-acetyl-2-deoxy-b-d-glucopyrano-
side (34).ÐA solution of 33 (40 mg, 0.036 mmol)
and DDQ (41 mg, 0.18 mmol) in 18:1 CH2Cl2±H2O
(10 mL) was vigorously stirred for 7 h at room
temperature. The reaction was quenched by addi-
tion of aq 20% Na2S2O3 (2.5 mL), the solution was
extracted with EtOAc, the organic layer was dried
(Na2SO4), and concentrated. The residue was pur-
i®ed by ¯ash-chromatogrphy on silica gel (24:1
EtOAc±MeOH). The product obtained (28.5 mg)
was dissolved in pyridine (6 mL) and treated with

J. Bartek et al./Carbohydrate Research 308 (1998) 259±273
271
ꢀ
00 00
Ac2O (0.7 mL) for 24 h at 0 C. MeOH (2 mL) was
added and the solution was taken to dryness.
Chromatography of the residue on silica gel (1:5
toluene±EtOAc) gave 34 as a syrup. Yield: 30 mg
(dd, 1 H, J6b,5 5.5 Hz, H-6b), 4.31 (d, 1 H, J1 ,2
00 00
8.5 Hz, H-100), 4.28 (dd, 1 H, J6b ,5 6.5 Hz, H-6b00),
4.20 (ddd, 1 H, H-200), 4.31±4.23 (m, 1 H, OCH2),
4.06 (t, 1 H, J3,2=J3,4=7.0 Hz, H-3), 4.05±3.99 (m,
1 H, OCH2), 3.91 (m, 1 H, H-2), 3.84 (m, 2 H, J4,5
7.0 Hz, H-4,5), 3.67 (m, 1 H, H-5), 2.19, 2.15,
2.149, 2.094, 2.087, 2.08, 2.014, 2.01, and 1.97 (9 s,
(96%); [ꢁ]20
91ꢀ (c 0.2, CHCl3). 1H NMR
d
(CDCl3): ꢂ 7.08 (d, 1 H, JNH ,2 9.0 Hz, NH00), 5.90
(d, 1 H, JNH,2 9.0 Hz, NH), 5.83 (m, 1 H, CH=),
00 00
5.38 (dd, J4 ,3 3.5, J4 ,5 0.8 Hz, H-400), 5.29 (d, 1
each 3 H, 9 Ac), 1.24 (d, 3 H, J6 ,5 6.6 Hz, H-60).
Anal. Calcd for C39H56N2O22: C, 51.76; H, 6.24;
N, 3.09. Found: C, 52.25; H, 6.08; N, 3.14.
00 00
00 00
0
0
H, J1 ,2 3.5 Hz, H-10), 5.29±5.22 (m, 1 H,
=CH2trans), 5.19±5.12 (m, 1 H, =CH2cis), 5.14 (dd,
0
0
1 H, J3 ,2 11.5 Hz, H-300), 4.84 (dd, 1 H, J2 ,3
Allyl (a-l-fucopyranosyl)-(1!3)-[(2-acetamido-
2-deoxy-b-d-galactopyranosyl)-(1!4)]-2-acetamido-
2-deoxy-b-d-glucopyranoside (36).ÐA solution of
35 (9.0 mg, 0.010 mmol) in MeOH (2 mL) was stir-
red with 0.1 M methanolic NaOMe (0.8 mL) for
24 h at ambient temperature. Dowex (H+) resin
was added to adjust the pH of the solution to 7.0.
The resin was ®ltered o€, and the ®ltrate was con-
centrated to give 6.0 mg (99%) of 36 as amorphous
69ꢀ (c 0.6, H2O). 1H NMR
(500 MHz, D2O): ꢂ 5.84 (m, 1 H, CH=), 5.24 (dq,
1 H, =CH2trans), 5.20 (dq, 1 H, =CH2cis), 5.05 (d,
1 H, J1 ,2 4.0 Hz, H-10), 4.79 (m, 1 H, H-50), 4.49
00 00
0
0
8.0 Hz, H-20), 4.58±4.56 (m, 1 H, H-50), 4.56 (d, 1 H,
00
00 00
H-1), 4.47 (d, 1 H, J1 ,2 8.0 Hz, H-1 ), 4.43 (dd, 1 H,
J6a,5 5.5, J6a,6b 12.0 Hz, H-6a), 4.33 (dd, 1 H, J6b,5
4.0 Hz, H-6b), 4.30 (dd, 1 H, J3 ,4 5.0 Hz, H-30),
0
0
4.29±4.21 (m, 3 H, H-200,6a00, and OCH2), 4.15 (dd,
1 H, J6b ,6a 11.0 Hz, H-6b00), 4.19±4.14 (m, 1 H, H-
00
00
40), 4.04±3.96 (m, 3 H, H-2,3, and OCH2), 3.90 (td,
1 H, J5 ,6a 6.5 Hz, H-500), 3.83±3.79 (m, 1 H, H-4),
3.72±3.68 (m, 1 H, H-5), 2.19, 2.13, 2.128, 2.08,
2.02, and 2.01 (6 s, each 3 H, 6 Ac), 1.52 and 1.36
00
00
solid; [ꢁ]20
d
0
0
(2 s, each 3 H, Me2C), 1.42 (d, 3 H, J6 ,5 6.7 Hz, H-
60). Anal. Calcd for C38H53Cl3N2O20: C, 47.33; H,
5.54; N, 2.90. Found: C, 46.86; H, 5.21; N, 2.75.
Allyl (2,3,4-tri-O-acetyl-a-l-fucopyranosyl)-
(1!3)-[(2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-b-
d-galactopyranosyl)-(1!4)]-2-acetamido-6-O-acetyl-
2-deoxy-b-d-glucopyranoside (35).ÐA solution of
34 (25 mg, 0.026 mmol) in CH2Cl2 (10 mL) was
treated with aq 90% CF3CO2H (0.2 mL) at 20 ꢀC
for 2 h. The solution was evaporated to dryness.
The residue was dissolved in EtOH (8 mL) and
stirred with NaBH4 (51 mg, 1.35 mmol) for 24 h at
60 ꢀC. After addition of EtOH (10 mL) the solution
0
0
00 00
(d, 1 H, J1,2 8.4 Hz, H-1), 4.40 (d, 1 H, J1 ,2 8.4 Hz,
H-100), 4.26 (ddt, 1 H, OCH2), 4.08 (ddt, 1 H,
OCH2), 3.91 (dd, 1 H, J2 ,3 10.8 Hz, H-200), 3.88
00 00
(dd, 1 H, J3 ,4 3.3 Hz, H-30), 3.87 (dd, 1 H, J2,3
12.2 Hz, H-2), 3.88±3.85 (m, 2 H, H-6a,400), 3.82 (t,
1 H, J4,3=J4,5=9.0 Hz, H-4), 3.78 (t, 1 H, H-3),
0
0
3.78 (m, 1 H, H-40), 3.71 (dd, 1 H, J6a ,5 7.8,
J6a ,6b 11.6 Hz, H-6a00), 3.68 (dd, 1 H, J6b,5 5.2,
00 00
00
00
00 00
J6a,6b 12.3 Hz, H-6b), 3.68 (dd, 1 H, J6b ,5 4.4 Hz,
00 00
H-6b00), 3.66 (dd, 1 H, J3 ,4 3.3 Hz, H-3 ), 3.64 (dd,
00
1 H, J2 ,3 10.1 Hz, H-20), 3.52 (dd, 1 H, J5 ,4
<1.0 Hz, H-500), 3.45 (ddd, 1 H, J5,6a 2.3 Hz, H-5),
1.99 and 1.96 (2 s, each 3 H, 2 Ac), 1.20 (d, 3 H,
0
0
00 00
ꢀ
was cooled to 0 C and HOAc (0.076 mL) was
added. The solution was coevaporated 5 times with
MeOH (10 mL). The residue was dissolved in pyr-
idine (8 mL) and stirred with Ac2O (4 mL) for 96 h
at 0 ꢀC. MeOH (5 mL) was added and the solution
was concentrated. Puri®cation of the residue on
silica gel (24:1 EtOAc±MeOH) furnished 14 mg
J6 ,5 6.6 Hz, H-60). ES-MS (C25H42N2O15): m/z
611.3 (M+H)+.
0
0
3-(2-Aminoethylthio)propyl (a-l-fucopyranosyl)-
(1!3)-[(2-acetamido-2-deoxy-b-d-galactopyano-
syl)-(1!4)]-2-acetamido-2-deoxy-b-d-glucopyrano-
side (37).ÐA solution of 36 (5.0 mg, 8.2 mmol) and
cysteamine hydrochloride (7.0 mg, 0.62 mmol) in
water (0.2 mL) was irradiated at 254 nm for 6 h at
room temperature. The solution was passed
through a column of Dowex AGWX8 resin
(60%) of 35 as syrup; [ꢁ]20
76ꢀ (c 0.3, MeOH).
d
1H NMR (CDCl3): ꢂ 5.95 (d, 1 H, JNH,2 9.0 Hz,
NH), 5.88 (d, 1 H, JNH ,2 9.0 Hz, NH00), 5.85 (m, 1
00 00
H, CH=), 5.41 (d, 1 H, J1 ,2 4.0 Hz, H-10), 5.37±
5.36 (m, 2 H, H-40,400), 5.30±5.22 (m, 1 H,
=CH2trans), 5.20±5.36 (m, 1 H, 0=CH2cis), 5.20 (dd,
0
0
+
(NH4 form) using 0.1 M aq NH3 as eluent. Car-
0
0
0
0
1 H, J3 ,2 11.0, J3 ,4 3.5 Hz, H-3 ), 5.06 (dd, 1 H, H-
bohydrate-containing fractions were pooled, lyo-
philized and further puri®ed on Sephadex G-10
(0.05 M aq NH4Ac) to give, after lyophilization, 37
20), 4.98 (dd, 1 H, J3 ,2 11.0, J3 ,4 3.5 Hz, H-300),
4.69 (m, 1 H, H-50), 4.60 (d, 1 H, J1,2 6.0 Hz, H-1),
4.45 (dd, 1 H, J6a,5 3.5, J6a,6b 11.5 Hz, H-6a), 4.41
00 00
00 00
as amorphous powder. Yield: 5.9 mg (96%); [ꢁ]20
d
(dd, 1 H, J6a ,5 6.5, J6a ,6b 11.5 Hz, H-6a00), 4.32
46ꢀ (c 0.5, H2O). 1H NMR (D2O): ꢂ 5.10 (d, 1 H,
00 00
00
00

272
J. Bartek et al./Carbohydrate Research 308 (1998) 259±273
J1 ,2 3.9 Hz, H-10), 4.83 (br d, 1 H, H-50), 4.48 (d, 1
References
0
0
00 00
H, J1,2 8.0 Hz, H-1), 4.45 (d, 1 H, J1 ,2 8.3 Hz, H-
100), 3.99±3.82 (m, 8 H, H-2,6a,30,40,200,300,400, and
OCH2), 3.80±3.62 (m, 7 H, H-3,4,6b,20,6a00,6b00,
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00
00
00
00
and OCH2), 3.56 (dd, 1 H, J5 ,6a 4.5, J5 ,6b 7.6 Hz,
H-500), 3.53±3.48 (m, 1 H, H-5), 3.19 (t, 2 H,
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2.04 and 2.03 (2 s, each 3 H, 2 Ac), 1.84 (br t, 2 H,
CH2), 1.25 (d, 3 H, J6 ,5 6.6 Hz, H-60). 13C NMR
(D2O): ꢂ 175.73 (C=O), 175.01 (C=O), 101.94 (C-
100), 101.63 (C-1), 99.32 (C-10), 76.34 (C-5), 75.73
(C-500), 75.55 (C-3), 74.26 (C-4), 72.89 (C-40), 71.62
(C-300), 70.08 (OCH2), 69.57 (C-30), 68.61 (C-20),
68.25 (C-400), 67.82 (C-50), 62.34 (C-600), 60.92 (C-
6), 56.55 (C-2), 53.27 (C-200), 39.55 (CH2N), 30.40
and 29.41 (2 CH2S), 27.97 (CH2), 23.18 and 23.08
(2 CH3), 16.25 (C-60). ES-MS (C27H49N3O15S): m/z
688.3 (M+H)+.
0
0
Synthesis of BSA-conjugate 38.ÐA solution of
thiophosgene (1 mL, 13 mmol) in CHCl3 (1 mL) was
added to a solution of compound 37 (3.7 mg,
5 mmol) in 0.1 M aq NaHCO3 (1.5 mL) and the
mixture was vigorously stirred for 3 h at room
temperature. The organic phase was separated and
the aqueous phase was washed three times with
CHCl3 (2 mL portions) and ®nally purged with N2
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a
Sephadex G-25 column
(1.6Â50 cm, 0.01 M aq. NaHCO3). Ninhydrin-
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The authors gratefully acknowledge ®nancial
support by the Austrian Science Foundation
(FWF-grant P 9038 MOB) and the Jubilaumsfonds
der Osterreichischen Nationalbank (Project
N5427). Furthermore, the authors thank Frank
Unger for helpful discussions, Andreas Ho®nger
for recording NMR spectra and Thomas Dalik for
technical assistance. Finally, accessibility and use
of NMR and mass spectrometry facilities at the
Department of Organic Chemistry, Eidgenossische
Technische Hochschule Zurich (Group of Pro-
fessor Diederich) is gratefully acknowledged.

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