Synthesis of the putative minimal FGF binding motif heparan sulfate trisaccharides by an orthogonal protecting group strategy

Article abstract of DOI:10.1016/j.tet.2008.08.021

The synthesis of two trisaccharides, the putative minimal heparan sulfate sequences responsible for binding to acidic and basic fibroblast growth factors, respectively, is described from a common protected intermediate using an orthogonal protecting group strategy.

Full text of DOI:10.1016/j.tet.2008.08.021

Tetrahedron 64 (2008) 9865–9873
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Synthesis of the putative minimal FGF binding motif heparan sulfate
trisaccharides by an orthogonal protecting group strategyq,qq
*
´
´
¨
Janos Tatai, Peter Fugedi
Department of Carbohydrate Chemistry, Chemical Research Center, Hungarian Academy of Sciences, Pusztaszeri u´t 59-67, H-1025 Budapest, Hungary
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 16 May 2008
Received in revised form 18 July 2008
Accepted 7 August 2008
Available online 12 August 2008
The synthesis of two trisaccharides, the putative minimal heparan sulfate sequences responsible for
binding to acidic and basic fibroblast growth factors, respectively, is described from a common protected
intermediate using an orthogonal protecting group strategy.
Ó 2008 Elsevier Ltd. All rights reserved.
Keywords:
Oligosaccharide synthesis
Heparin
Heparan sulfate
Fibroblast growth factors
Binding epitope
Orthogonal protection
1. Introduction
oligosaccharide sequences with a unique sulfation pattern are re-
quired to mediate the activities of individual heparin-binding
Heparin (H) and heparan sulfate (HS) are sulfated glycosami-
noglycans (GAGs) built up of linear chains of alternating -glucos-
amine and hexuronic acid units.1–4 The carbohydrate backbone,
consisting of /4)- -IdopA-(1/4)- -GlcpN-(1/ and /4)-
-GlcpA-(1/4)- -GlcpN-(1/ disaccharide units, is substituted
at certain positions. The variations of the hexuronic acid units (
iduronic or -glucuronic acid), the substitutions of the amino
groups (with acetyl or sulfate groups), and the sulfations of hy-
droxyl groups (O-3 and O-6 of the -glucosamine, O-2 of the uronic
proteins.5,9
D
Fibroblast growth factors (FGFs) are a family of more than 20
structurally related signaling polypeptides involved in processes
such as cell proliferation, differentiation, and migration.10–12 The
biological activities of FGFs are tightly regulated by heparin and
heparan sulfate.13–16 Heparan sulfate interacts both with FGFs and
their cell surface receptors (FGFRs); FGF signaling requires the
formation of ternary complex between FGF, FGFR, and HS.17 The
structural requirements of heparin and heparan sulfate to modu-
late FGF signaling are intensely studied, and a great deal of in-
formation in relation to the best studied acidic (FGF-1) and basic
fibroblast growth factors (FGF-2) has emerged.13,14 It was de-
termined that FGF-1 and FGF-2 have distinct O-sulfation re-
quirements for activation:18 both 2-O-sulfation [IdopA(2-OSO3)]
and 6-O-sulfation [GlcpN(6-OSO3)] are needed for binding to FGF-
1,19 whereas FGF-2 requires 2-O-sulfation [IdopA(2-OSO3)] only, 6-
O-sulfation is not essential for binding.20 In addition to the gross
structural features of H/HS required for FGF-1 and FGF-2 binding,
a number of different oligosaccharide structures having high af-
finity for these FGFs have been proposed.20–30 Recently, the hep-
aran sulfate sequence 1 has been identified as the minimal binding
motif for FGF-1, and sequence 2 as a high affinity binding epitope
for FGF-2 (Fig. 1).31
a
-L
a
-D
b-
D
a-D
L-
D
D
acids) result in an enormous structural diversity. Heparin and
heparan sulfate interact with a large number of proteins including
enzymes, protease inhibitors, growth factors, chemokines, and
several others.5–8 These glycosaminoglycan/protein interactions
regulate
a series of important biological processes. Though
the binding of heparin and heparan sulfate to some proteins is
nonspecific, simply ionic by nature, many of the biologically rele-
vant interactions of H/HS show a great deal of specificity. It is
commonly assumed, though not rigorously proven, that specific
q
Synthesis of Glycosaminoglycan Oligosaccharides, Part 2. For Part 1 see, Ref. 47.
Presented in part at the 22nd International Carbohydrate Symposium,
qq
Glasgow, United Kingdom, July 23–27, 2004; Abstract P204; and at the 13th
European Carbohydrate Symposium, Bratislava, Slovakia, August 21–26, 2005;
Abstract P56.
* Corresponding author. Tel.: þ36 1 438 1100; fax: þ36 1 438 1145.
E-mail address: pfugedi@chemres.hu (P. Fu¨ gedi).
These structures were deduced from the study of a heparan
sulfate octasaccharide library, the actual trisaccharides were not
tested as they were not available. The trisaccharides 1 and 2, with
0040-4020/$ – see front matter Ó 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.tet.2008.08.021

9866
J. Tatai, P. Fu¨gedi / Tetrahedron 64 (2008) 9865–9873
NaO2C
2. Results and discussion
O
OH
O
O
NaO3SO
O
2.1. Retrosynthesis of the trisaccharides
NH
O
OH
HO
OSO3Na
SO3Na
NaO2C
O
The target trisaccharides (3 and 4) share the same carbohydrate
backbone and differ only in their sulfation pattern. Retrosynthesis
of 3 and 4 (Scheme 1) revealed that both compounds could be
accessed from a single common intermediate, if the O-2 positions
1
OSO3Na
O
NaO2C
O
O
OH
O
of the
are protected by orthogonal protecting groups. The benzoyl group
was selected for the O-2 protection of the -iduronic acids, due to its
advantageous properties in glycosylation reactions45 and the (4-
methoxy)phenyl (MPh) group46 for the protection of O-6 of the
L-iduronic acid units and O-6 of the D-glucosamine residue
HO
O
NH
L
O
HO
OSO3Na
SO3Na
OH
O
NaO2C
D
-
2
OSO3Na
glucosamine residue. The latter group, which can be removed with
ceric ammonium nitrate (CAN) similarly to the commonly used 4-
methoxybenzyl residue, was preferred due to its more robust, acid-
resistant character. We have shown previously43,44 that these
groups are not only orthogonal, they are also stable under O-sul-
fations and can be selectively removed without affecting the sen-
sitive O-sulfate groups. The orthogonally protected trisaccharide 5
should be available from the monosaccharide building blocks 6, 7,
and 8.
O
Figure 1. Minimal heparan sulfate binding motifs of FGF-1 and FGF-2.
L
-iduronic acid at the reducing end, are not obtainable by
chemical or enzymatic degradation of heparin, as the current
methods afford only oligosaccharides with the amino sugar (or its
degradation product) at the reducing end. The chemical synthesis
of the suggested trisaccharide ligands has not been accomplished
yet.
The chemical synthesis of H/HS oligosaccharides is intensely
studied32–36 and a number of oligosaccharides have been syn-
thesized in relation to FGFs by different groups.37–42 Syntheses
of heparin oligosaccharides are elaborate, multi-step processes,
and a common feature of the previous syntheses is that they
afford a single heparin oligosaccharide after a long synthetic
sequence.
2.2. Synthesis of the
L-iduronic acid building blocks
We have recently described47 an efficient synthesis for the non-
reducing end L-iduronic acid thioglycoside 6. For the synthesis of 8
(Scheme 2), the thioglycoside 947 was chosen as starting material.
Glycosylation of 9 with methanol in dichloromethane in the pres-
ence of dimethyl(methylthio)sulfonium triflate45 (DMTST) afforded
In a program on the synthesis of glycosaminoglycan oligosac-
charides, we have previously developed43,44 a synthesis strategy
based on orthogonal protection, which allows the synthesis of
multiple sulfated oligosaccharides from a single protected pre-
an anomeric mixture of the methyl glycosides, from which the
glycoside 10 was obtained in 49% yield. A significant amount (35%)
of the -glycoside was also isolated. Although various -iduronic
acid donors generally give high stereoselectivity in reactions with
complex glycosyl acceptors in oligosaccharide syntheses, formation
a-
b
L
a
cursor. We now report on the synthesis of the methyl a-glycosides
(3 and 4) of the putative FGF binding trisaccharides (1 and 2) using
this orthogonal protection-based methodology.
of anomeric mixtures with a high proportion of the b-glycoside are
not unprecedented in glycosylations of simple alcohols48–50 such as
NaO2C
OH
O
OMe
O
RO
O
NH
SO3Na
O
HO
OSO3Na
OH
O
NaO2C
3 R=SO3Na
4 R=H
OSO3Na
OH
t-BuO2C
OBn
OMe
O
O
MPhO
O
N3
O
BnO
OBz
OBn
O
t-BuO2C
5
OBz
OBn
OMe
SPh
OMPh
O
OBn
O
OBn
O
ClAcO
BnO
t-BuO2C
t-BuO2C
BnO
SPh
N3
OH OBz
8
OBz
6
7
Scheme 1. Retrosynthesis of trisaccharides 3 and 4.

J. Tatai, P. Fu¨gedi / Tetrahedron 64 (2008) 9865–9873
9867
methanol or isopropyl alcohol. Regioselective reductive opening of
2.4. Synthesis of the orthogonally protected trisaccharide
DMTST-promoted glycosylation45 of the
-iduronic acid acceptor
the (1-naphthyl)methylene acetal 10 with BH3$THF/TMSOTf51 gave
the corresponding 4-O-(1-naphthyl)methyl derivative 11. Oxidation
of 11 using pyridinium dichromate and acetic anhydride in the
L
8 with the -glucosamine thioglycoside 7 resulted stereoselectively
D
presence of tert-butanol52 afforded the tert-butyl uronate 12. The
L-
in the -linked disaccharide 19 (Scheme 4). Removal of the tem-
a
iduronic acid glycosyl acceptor 8 was obtained by oxidative re-
moval of the (1-naphthyl)methyl group with CAN to give 8 in 76%
yield.
porary chloroacetyl protecting group by hydrazinedithiocar-
bonate54 afforded the alcohol 20. This disaccharide acceptor (20)
was coupled with the L-iduronic acid thioglycoside 6 in the pres-
ence of DMTST to give the orthogonally protected trisaccharide 5 in
61% yield.
SPh
OMe
OBn
O
OBn
O
b
d
a
c
t-BuO2C
O
O
OBn
O
OMe
O
OBz
9
O
OBz
10
O
MPhO
RO
1Naph
1Naph
O
a
N3
7
+
8
BnO
OBz
19 R=AcCl
20 R=H
b
OMe
OMe
OBn
O
OBn
O
HO
1NAPO
t-BuO2C
t-BuO2C
SPh
OBn
O
OMe
OBn
O
O
1NAPO
OBz
12
c
OBz
11
MPhO
HO
+
t-BuO2C
O
N3
20
BnO
OBz
OBn OBz
6
OMe
OBn
t-BuO2C
O
OBn
t-BuO2C
OMe
O
O
MPhO
O
OH OBz
8
N3
O BnO
OBz
OBn
O
Scheme 2. Synthesis of L-iduronic acid glycosyl acceptor 8; (a) DMTST, MeOH, CH2Cl2,
49%; (b) BH3$THF, TMSOTf, CH2Cl2, 66%; (c) PDC, Ac2O, t-BuOH, CH2Cl2, 67%; (d) CAN,
MeCN, H2O, 76%.
t-BuO2C
5
OBz
OBn
Scheme 4. Synthesis of the orthogonally protected trisaccharide 5; (a) DMTST, Et2O,
CH2Cl2, 47%; (b) HDTC, DMF, 80%; (c) DMTST, CH2Cl2, 61%.
2.3. Synthesis of the D-glucosamine building block
2.5. Preparation of heparan sulfate trisaccharides
The known triol 1353 was reacted with 1-naphthaldehyde di-
methyl acetal in DMF solution in the presence of camphorsulfonic
acid to afford 14 (Scheme 3). After benzylation of the free hydroxyl
group, acid hydrolysis of the (1-naphthyl)methylene acetal of 15 led
to 16. Regioselective tosylation of the primary hydroxyl group of 16
and nucleophilic replacement of the resulting tosylate 17 with so-
dium (4-methoxy)phenoxide gave the 6-O-(4-methoxy)phenyl
derivative 18 in 83% yield. Chloroacetylation of 18 then afforded the
´
Debenzoylation of compound 5 (Scheme 5) using mild Zemplen
deacylation prevented possible -elimination of the base-sensitive
-iduronic residues. Though the reaction was sluggish due to the
b
L
isolated nature of the benzoyl groups55 it provided the diol 21 in
high yield. Cleavage of the (4-methoxy)phenyl group by treatment
with CAN afforded triol 22. After ester hydrolysis, the free acid 23
was sulfated with sulfur trioxide pyridine complex in DMF and the
product was converted into pentasodium salt (24) by Dowex 50
(Naþ form) resin. Catalytic hydrogenation with palladium on car-
bon removed the benzyl groups and reduced the azido group.
Chemoselective N-sulfation of 25 using sulfur trioxide trimethyl-
amine complex in water at pH 9.8 afforded the putative FGF-1
binding trisaccharide (3).
A different sequence of deprotection and sulfation steps starting
from the same orthogonally protected trisaccharide (5) also affor-
ded the putative FGF-2 binding trisaccharide (4). First the tert-butyl
esters were cleaved from the debenzoylated trisaccharide 21 to give
26. The dihydroxy uronic acid, still having the (4-methoxy)phenyl
group, was first sulfated, and the (4-methoxy)phenyl group was
removed from 27 by treatment with CAN to give 28 having the
sulfate esters intact (28). Hydrogenolysis followed by N-sulfation of
the unprotected trisaccharide 29 resulted in the putative FGF-2
binding trisaccharide (4).
D-glucosamine building block 7.
OH
1Naph
O
O
O
a
b
HO
HO
O
SPh
SPh
SPh
HO
N3
13
N3
14
OH
O
1Naph
O
O
HO
c
O
d
f
SPh
BnO
BnO
N3
16
N3
15
OMPh
O
OTs
O
HO
e
HO
BnO
SPh
SPh
BnO
N3
18
N3
17
3. Conclusion
OMPh
O
ClAcO
BnO
SPh
In summary, we have synthesized the heparan sulfate tri-
saccharides 3 and 4, the putative minimal binding epitopes of FGF-
1 and FGF-2, respectively. Both target compounds were readily
prepared from the same orthogonally protected trisaccharide 5.
The synthesis relies on a new pair of orthogonal protecting groups
[benzoyl and (4-methoxy)phenyl], which, in addition to their
N3
7
Scheme 3. Synthesis of the D-glucosamine building block 7; (a) 1NaphCH(OMe)2, CSA,
DMF, 75%; (b) BnBr, NaH, DMF, 86%; (c) 60% aq AcOH, 90%; (d) TsCl, DMAP, Et3N,
CH2Cl2; (e) 4-methoxyphenol, NaH, DMF, 83% for two steps; (f) (ClCH2CO)2O, pyridine,
CH2Cl2, 81%.

9868
J. Tatai, P. Fu¨gedi / Tetrahedron 64 (2008) 9865–9873
t-BuO2C
OBn
O
OMe
O
MPhO
O
N3
21
a
O
BnO
OH
5
OBn
O
t-BuO2C
OH
OBn
c
b
R3O2C
R3O2C
OBn
O
OBn
OMe
OMe
O
O
O
R2O
R2O
O
O
N3
OR1
N3
OR1
O
BnO
O
BnO
OBn
O
OBn
O
R3O2C
R3O2C
OR1
OR1
OBn
OBn
26 R1=H
R2=MPh
R3=H
22 R1=H
23 R1=H
R2=H
R2=H
R3=t-Bu
c
d
b
27 R1=SO3Na R2=MPh
28 R1=SO3Na R2=H
R3=Na
R3=Na
R3=H
24 R1=SO3Na R2=SO3Na R3=Na
d
e
e
NaO2C
NaO2C
OH
OH
O
OMe
OMe
O
O
O
NaO3SO
HO
O
O
NH
NH
O
HO
OSO3Na
O
HO
OSO3Na
R
R
OH
O
OH
O
NaO2C
NaO2C
OSO3Na
f
OH
OSO3Na
f
OH
25 R=H
29 R=H
3
R=SO3Na
4
R=SO3Na
Scheme 5. Synthesis of the putative FGF binding trisaccharides 3 and 4; (a) NaOMe, MeOH 93%; (b) CAN, MeCN, H2O, 84% for 22, 83% for 28; (c) CF3CO2H, CH2Cl2, 98% for 23, 92% for
26; (d) Py$SO3, DMF, 68% for 24, 99% for 27; (e) Pd/C, THF, H2O; (f) NMe3$SO3, NaOH, H2O, 45% for two steps for 3, 68% for two steps for 4.
orthogonality, can also be removed selectively in the presence of O-
sulfate groups.
Analytical Department of the Chemical Research Center, Hungarian
Academy of Sciences.
The synthesis also demonstrates that the preparation of multi-
ple heparin/heparan sulfate oligosaccharides is more efficient and
less time consuming by our orthogonal protection strategy, than
synthesizing these oligosaccharides individually.
4.2. Methyl 2-O-benzoyl-3-O-benzyl-4,6-O-(1-naphthyl)-
methylidene-a-L-idopyranoside (10)
A mixture of 947 (4.0 g, 6.6 mmol), MeOH (2.6 mL, 66.0 mmol),
and 4 Å molecular sieves (3 g) was stirred in dry CH2Cl2 (30 mL)
under argon for 30 min, then DMTST (5.1 g, 19.8 mmol) was added.
After 1 day, the reaction was quenched with triethylamine (5 mL).
The mixture was filtered through a pad of Celite, the filtrate was
diluted with CH2Cl2 (500 mL) and washed with 2 M aq HCl (100 mL),
saturated aq NaHCO3 (100 mL), and water (100 mL), dried, and
concentrated. The residue was purified by column chromatography
(95:5 toluene/EtOAc) to give first 10 (1.7 g, 49%); mp 140–141 ꢀC
4. Experimental
4.1. General methods
Organic solutions were dried over MgSO4 and concentrated
under reduced pressure at 40 ꢀC. Thin-layer chromatography (TLC)
was performed on Silica gel 60 F254 plates (E. Merck, Darmstadt),
the compounds were detected under UV light and by spraying the
plates with a 0.02 M solution of resorcinol in 20% methanolic H2SO4
solution followed by heating. For column chromatography, silica gel
60 (0.040–0.063 mm) (E. Merck) was employed. Melting points
were determined in capillary tubes on a Griffin melting point ap-
paratus and are uncorrected. Optical rotations were measured at
23 ꢀC with an Optical Activity AA-10R polarimeter. The NMR
spectra were recorded on a Varian Gemini 2000 (1H: 200 MHz; 13C:
50 MHz) and Varian Unity-Inova (1H: 400 MHz; 13C: 100 MHz)
spectrometers at ambient temperature. The chemical shifts were
referenced to TMS (0.00 ppm for 1H) and to the central line of CDCl3
(77.16 ppm for 13C) for solutions in CDCl3, to the central line of the
solvent (3.31 ppm for 1H, 49.00 ppm for 13C) for solutions in CD3OD,
and to the methyl signal of acetone (2.22 ppm for 1H, 30.89 ppm for
13C) for solutions in D2O, as internal standards. Elemental analyses
were performed with an Elementar Vario EL III instrument at the
(from EtOAc/hexanes); [
CDCl3):
a
]
D ꢁ102 (c 0.4, CHCl3); 1H NMR (400 MHz,
d
8.42 (m, 1H, aromatic), 7.87–7.80 (m, 4H, aromatic), 7.71
(dd, 1H, aromatic), 7.45–7.24 (m, 9H, aromatic), 7.11–7.06 (m, 2H,
aromatic), 6.06 (s,1H,1NaphCH), 5.29 (ddd,1H, J1,2 1.7 Hz, J2,3 2.9 Hz,
J2,4 0.8 Hz, H-2), 5.06 (d, 1H, J1,2 1.7 Hz, H-1), 4.88 (d, 1H, J 12.1 Hz,
PhCH2), 4.70 (d, 1H, J 12.1 Hz, PhCH2), 4.43 (dd, 1H, J5,6a 1.6 Hz, J6a,6b
12.5 Hz, H-6a), 4.25 (dd, 1H, J5,6b 1.8 Hz, J6a,6b 12.5 Hz, H-6b), 4.16
(ddd, 1H, J2,4 0.8 Hz, J3,4 2.5 Hz, J4,5 1.5 Hz, H-4), 4.09 (ddd, 1H, J4,5
1.5 Hz, J5,6a 1.6 Hz, J5,6b 1.8 Hz, H-5), 3.81 (dd, 1H, J2,3 2.9 Hz, J3,4
2.5 Hz, H-3), 3.52 (s, 3H, OCH3); 13C NMR (100 MHz, CDCl3):
d 165.8
(OC(O)Ph), 137.7, 133.9, 133.4, 133.0, 130.6, 130.1, 129.7, 129.5, 128.5,
128.3, 128.1, 127.92, 127.88, 126.3, 125.6, 125.0, 124.93, 124.86 (aro-
matic),101.2 (1NaphCH),100.0 (C-1, JC-1,H-1168.4 Hz), 74.6 (C-3), 74.1
(C-4), 72.1 (PhCH2), 70.0 (C-6), 66.8 (C-2), 59.8 (C-5), 55.8 (OCH3).
Anal. Calcd for C32H30O7: C, 72.99; H, 5.74. Found: C, 72.76; H, 5.79.

J. Tatai, P. Fu¨gedi / Tetrahedron 64 (2008) 9865–9873
9869
Eluted second was the
b
-anomer, methyl 2-O-benzoyl-3-O-
127.9, 126.2, 126.1, 125.7, 125.1, 123.7 (aromatic), 99.8 (C-1), 82.0
(C(CH3)3), 75.2 (C-4), 73.5 (C-3), 72.6, 70.9 (2ArCH2), 68.9 (2C, C-
2,5), 56.2 (OCH3), 28.1 (C(CH3)3). Anal. Calcd for C36H38O8: C, 72.22;
H, 6.40. Found: C, 72.03; H, 6.41.
benzyl-4,6-O-(1-naphthyl)methylidene-b-L-idopyranoside (1.2 g,
35%) as a syrup; [
8.20 (m, 1H, aromatic), 8.01–7.95 (m, 2H, aromatic), 7.85–7.70 (m,
a
]
D þ40 (c 0.4, CHCl3); 1H NMR (400 MHz, CDCl3):
d
3H, aromatic), 7.52–7.34 (m, 8H, aromatic), 7.25–7.09 (m, 3H, aro-
matic), 6.09 (s, 1H, 1NaphCH), 5.35 (ddd, 1H, J1,2 1.6 Hz, J2,3 2.8 Hz,
J2,4 0.6 Hz, H-2), 4.92 (d, 1H, J1,2 1.6 Hz, H-1), 4.86 (d, 1H, J 12.1 Hz,
PhCH2), 4.73 (d, 1H, J 12.1 Hz, PhCH2), 4.52 (dd, 1H, J5,6a 2.0 Hz, J6a,6b
12.4 Hz, H-6a), 4.26 (dd, 1H, J5,6b 1.5 Hz, J6a,6b 12.4 Hz, H-6b), 4.09
(ddd, 1H, J2,4 0.6 Hz, J3,4 2.4 Hz, J4,5 1.5 Hz, H-4), 4.01 (dd, 1H, J2,3
2.8 Hz, J3,4 2.4 Hz, H-3), 3.87 (ddd, 1H, J4,5 1.5 Hz, J5,6a 2.0 Hz, J5,6b
4.5. tert-Butyl (methyl 2-O-benzoyl-3-O-benzyl-a-L-
idopyranoside)uronate (8)
To a solution of 12 (1.5 g, 2.4 mmol) in MeCN (20 mL) and water
(5 mL), CAN (4.0 g, 7.2 mmol) was added. The mixture was stirred at
room temperature for 3 h, neutralized with saturated aq NaHCO3
(10 mL), diluted with CH2Cl2 (500 mL), washed with saturated aq
NaHCO3 (150 mL) and water (150 mL), dried, and concentrated. The
residue was purified by column chromatography (98:2 toluene/
1.5 Hz, H-5), 3.60 (s, 3H, OCH3); 13C NMR (100 MHz, CDCl3):
d 166.5
(OC(O)Ph), 137.4, 133.7, 133.1, 132.8, 130.5, 130.3, 129.9, 129.5, 128.6,
128.4, 128.2, 128.0, 127.8, 126.3, 125.5, 124.9, 124.7, 124.2 (aromatic),
100.0 (1NaphCH), 99.2 (C-1, JC-1,H-1 156.8 Hz), 75.9 (C-3), 73.3 (C-4),
72.8 (PhCH2), 69.9 (C-6), 67.0 (C-5), 66.7 (C-2), 57.1 (OCH3). Anal.
Calcd for C32H30O7: C, 72.99; H, 5.74. Found: C, 72.75; H, 5.72.
EtOAc) to give 8 as a syrup (0.85 g, 76%); [
NMR (200 MHz, CDCl3):
a
]
D ꢁ17.1 (c 0.3, CHCl3); 1H
d
8.02–7.97 (m, 2H, aromatic), 7.63–7.27
(m, 8H, aromatic), 5.20 (ddd, 1H, J1,2 2.9 Hz, J2,3 3.2 Hz, J2,4 0.8 Hz, H-
2), 4.99 (dd, 1H, J1,2 2.9 Hz, J1,3 1.0 Hz, H-1), 4.86 (d, 1H, J 12.1 Hz,
PhCH2), 4.74 (d, 1H, J4,5 1.5 Hz, H-5), 4.66 (d, 1H, J 12.1 Hz, PhCH2),
4.02 (dddd, 1H, J2,4 0.8 Hz, J3,4 3.9 Hz, J4,5 1.5 Hz, J4,OH 11.4 Hz, H-4),
3.84 (ddd, 1H, J1,3 1.0 Hz, J2,3 3.2 Hz, J3,4 3.9 Hz, H-3), 3.51 (s, 3H,
OCH3), 2.75 (d, 1H, J4,OH 11.4 Hz, OH), 1.51 (s, 9H, C(CH3)3); 13C NMR
4.3. Methyl 2-O-benzoyl-3-O-benzyl-4-O-(1-naphthyl)-
methyl-a-L-idopyranoside (11)
To a solution of 10 (2.6 g, 4.9 mmol) in dry CH2Cl2 (50 mL), 1 M
BH3$THF in THF (9.8 mL, 9.8 mmol) and TMSOTf (0.09 mL,
0.49 mmol) were added. The mixture was stirred for 2 h at room
temperature, cooled to 0 ꢀC, and quenched with triethylamine
(5 mL) and MeOH (25 mL). The solution was concentrated in vacuo
and co-evaporated three times with MeOH (25 mL). The residue
was purified by column chromatography (98:2/9:1 toluene/ace-
(50 MHz, CDCl3):
d 168.6 (C-6), 165.1 (OC(O)Ph), 137.7, 133.7, 129.9,
129.2, 128.7, 128.6, 128.1, 128.0 (aromatic), 100.0 (C-1), 82.5
(C(CH3)3), 75.0 (C-3), 72.3 (PhCH2), 68.3, 67.7, 67.6 (C-2,4,5), 56.4
(OCH3), 28.2 (C(CH3)3). Anal. Calcd for C25H30O8: C, 65.49; H, 6.60.
Found: C, 65.33; H, 6.57.
tone) to give 11 (1.7 g, 66%) as a syrup; [
NMR (200 MHz, CDCl3):
a
]
ꢁ23 (c 0.4, CHCl3); 1H
4.6. Phenyl 2-azido-2-deoxy-4,6-O-(1-naphthyl)methylidene-
1-thio-b-D-glucopyranoside (14)
D
d
8.04–7.72 (m, 5H, aromatic), 7.57–7.08
(m, 12H, aromatic), 5.22 (ddd, 1H, J1,2 1.6 Hz, J2,3 2.9 Hz, J2,4 0.7 Hz,
H-2), 4.94 (d, 1H, J 11.7 Hz, ArCH2), 4.88 (d, 1H, J1,2 1.6 Hz, H-1), 4.83
(d, 1H, J 11.7 Hz, ArCH2), 4.68 (d, 1H, J 12.1 Hz, ArCH2), 4.62 (d, 1H, J
12.1 Hz, ArCH2), 4.23 (ddd, 1H, J4,5 2.8 Hz, J5,6a 4.4 Hz, J5,6b 6.6 Hz, H-
5), 3.96 (dd, 1H, J2,3 2.9 Hz, J3,4 3.0 Hz, H-3), 3.88 (dd, 1H, J5,6a 4.4 Hz,
J6a,6b 8.8 Hz, H-6a), 3.61 (dd, J5,6b 6.6 Hz, J6a,6b 8.8 Hz,1H, H-6b), 3.56
(ddd, 1H, J2,4 0.7 Hz, J3,4 3.0 Hz, J4,5 2.8 Hz, H-4), 3.44 (s, 3H, OCH3),
To a solution of 1353 (3.7 g, 12.5 mmol) in dry DMF (30 mL), 1-
naphthaldehyde dimethyl acetal (4.0 mL, 18.8 mmol), and
(ꢂ)-camphor-10-sulfonic acid (0.3 g, 1.3 mmol) were added. The
mixture was stirred at 50 ꢀC under reduced pressure (40 kPa) for
5 h, neutralized with saturated aq NaHCO3 (10 mL), and concen-
trated. To the residue, hexanes (100 mL) and water (100 mL) were
added, and the mixture was stirred overnight. The precipitated
crystalline material was filtered off and crystallized from EtOH to
1.96 (m, 1H, OH); 13C NMR (50 MHz, CDCl3):
d 165.7 (OC(O)Ph),
137.8, 133.8, 133.3, 132.8, 131.6, 130.1, 129.7, 129.1, 128.7, 128.6,128.5,
128.2, 128.1, 127.2, 126.5, 126.0, 125.2, 123.8 (aromatic), 99.7 (C-1),
73.8 (C-4), 72.4, 71.9 (2ArCH2), 70.5 (C-3), 68.7, 67.7 (C-2, C-5), 62.7
(C-6), 55.7 (OCH3). Anal. Calcd for C32H32O7: C, 72.71; H, 6.10.
Found: C, 72.53; H, 6.07.
afford 14 as awhitesolid (4.1 g, 75%); mp 135–136 ꢀC; [
a
]D ꢁ42 (c 0.5,
CHCl3); 1H NMR (200 MHz, CDCl3):
d 8.22–7.32 (m, 12H, aromatic),
5.99 (s, 1H, 1NaphCH), 4.40 (dd, 1H, J5,6a 4.8 Hz, J6a,6b 10.6 Hz, H-6a),
4.34 (d, 1H, J1,2 9.9 Hz, H-1), 3.81 (dd, 1H, J5,6b 10.3 Hz, J6a,6b 10.6 Hz,
H-6b), 3.65–3.30 (m, 4H, H-3, H-4, H-5, OH), 3.24 (dd,1H, J1,2 9.9 Hz,
4.4. tert-Butyl [methyl 2-O-benzoyl-3-O-benzyl-4-O-(1-
J2,3 8.8 Hz, H-2); 13C NMR (50 MHz, CDCl3):
d 134.0,133.8,132.1,131.1,
naphthyl)methyl-
a
-
L
-idopyranoside]uronate (12)
130.4, 130.3, 129.2, 128.9, 128.8, 126.5, 125.9, 125.1, 125.0, 124.2 (ar-
omatic), 101.6 (1NaphCH), 86.9 (C-1), 80.8 (C-4), 74.1 (C-3), 70.4 (C-
5), 68.8 (C-6), 65.2 (C-2). Anal. Calcd for C23H21N3O4S: C, 63.43; H,
4.86; N, 9.65; S, 7.36. Found: C, 63.16; H, 4.81; N, 9.62; S, 7.40.
To a solution of 11 (1.7 g, 3.3 mmol) in CH2Cl2 (40 mL) pyr-
idinium dichromate (2.5 g, 6.6 mmol), Ac2O (3.1 mL, 33.0 mmol),
and tert-butyl alcohol (6.2 mL, 66.0 mmol) were added. The mix-
ture was stirred for 6 h at room temperature and it was then ap-
plied on the top of a silica gel column in EtOAc, with a 10 cm layer of
EtOAc on top of the gel. The chromium compounds were allowed to
precipitate in the presence of EtOAc and after 30 min the product
was eluted with EtOAc. After evaporating the solvent, the residue
was purified by column chromatography (98:2 toluene/acetone) to
4.7. Phenyl 2-azido-3-O-benzyl-2-deoxy-4,6-O-(1-
naphthyl)methylidene-1-thio-b-D-glucopyranoside (15)
To a stirred solution of 14 (4.1 g, 9.2 mmol) in dry DMF (40 mL),
60% NaH suspension (0.75 g, 18.4 mmol) was added at 0 ꢀC. After
30 min, benzyl bromide (1.6 mL, 13.8 mmol) was added dropwise.
Stirring was continued for 2 h, then MeOH (2 mL) was added. After
30 min, the mixture was diluted with CH2Cl2 (300 mL), washed
with water (100 mL), dried, and concentrated. The residue was
crystallized from EtOAc/hexanes to afford 15 as a white solid (4.2 g,
give 12 as a syrup (1.3 g, 67%); [
(200 MHz, CDCl3):
a
]
ꢁ42 (c 0.3, CHCl3); 1H NMR
D
d
7.91–7.72 (m, 5H, aromatic), 7.47–7.14 (m, 12H,
aromatic), 5.14 (ddd, 1H, J1,2 2.8 Hz, J2,3 3.3 Hz, J2,4 0.8 Hz, H-2), 5.06
(dd, 1H, J1,2 2.8 Hz, J1,3 0.7 Hz, H-1), 4.96 (d, 1H, J 12.0 Hz, ArCH2),
4.89 (d, 1H, J 12.0 Hz, ArCH2), 4.78 (d, 1H, J 11.9 Hz ArCH2), 4.71 (d,
1H, J4,5 3.1 Hz, H-5), 4.57 (d, 1H, J 11.9 Hz ArCH2), 4.02 (ddd, 1H, J2,4
0.8 Hz, J3,4 3.9 Hz, J4,5 3.1 Hz, H-4), 3.90 (ddd, 1H, J1,3 0.7 Hz, J2,3
86%); mp 159–160 ꢀC; [
CDCl3):
a
]
D
ꢁ79 (c 0.3, CHCl3); 1H NMR (200 MHz,
d
8.12–7.23 (m, 17H, aromatic), 6.10 (s, 1H, 1NaphCH), 4.78
(d, 1H, J 11.0 Hz, PhCH2), 4.66 (d, 1H, J 11.0 Hz, PhCH2), 4.52 (d, 1H,
J1,2 10.3 Hz, H-1), 4.50 (dd, 1H, J5,6a 4.8 Hz, J6a,6b 10.6 Hz, H-6a), 3.90,
3.78, 3.66 (3H, 3 t, H-3,4,6b), 3.55 (dd, 1H, J4,5 9.5 Hz, J5,6a 4.8 Hz,
J5,6b 9.5 Hz, H-5), 3.39 (dd, 1H, J1,2 10.3 Hz, J2,3 8.8 Hz, H-2); 13C NMR
3.3 Hz, J3,4 3.9 Hz, H-3), 3.49 (s, 3H, OCH3), 1.41 (s, 9H, C(CH3)3); 13
NMR (50 MHz, CDCl3): 168.5 (C-6), 165.6 (OC(O)Ph), 137.8, 133.5,
133.2, 133.0, 131.3, 129.9, 129.6, 128.47, 128.45, 128.41, 128.2, 128.1,
C
d

9870
J. Tatai, P. Fu¨gedi / Tetrahedron 64 (2008) 9865–9873
(50 MHz, CDCl3):
d
137.5, 134.2, 133.9, 132.3, 130.7, 130.6, 130.0,
NaHCO3 (75 mL), and water (75 mL), dried, and concentrated. The
residue was purified by column chromatography (95:5 toluene/ac-
etone) to give 7 (1.5 g, 81%); mp 75–76 ꢀC (from EtOAc/hexanes);
129.3, 128.9, 128.8, 128.6, 128.5, 128.1, 126.5, 125.9, 125.2, 124.1,
124.0 (aromatic), 100.4 (1NaphCH), 86.8 (C-1), 81.9, 81.1 (C-3,4),
75.4 (PhCH2), 70.7 (C-5), 68.9 (C-6), 64.9 (C-2). Anal. Calcd for
[a
]
D ꢁ35 (c 0.3, CHCl3); 1H NMR (200 MHz, CDCl3):
d 7.60–6.80 (m,
C
30H27N3O4S: C, 68.55; H, 5.18; N, 7.99; S, 6.10. Found: C, 68.09; H,
14H, aromatic), 5.09 (dd,1H, J3,4 9.1 Hz, J4,5 9.5 Hz, H-4), 4.87 (d,1H, J
11.7 Hz, PhCH2), 4.64 (d,1H, J 11.7 Hz, PhCH2), 4.49 (d,1H, J1,2 9.9 Hz,
H-1), 4.07–3.93 (m, 2H, H-6a, H-6b), 3.76 (m, 1H, H-5), 3.75 (s, 1H,
OCH3), 3.72 (d, 1H, J 7.3 Hz, C(O)CH2Cl), 3.57 (dd, 1H, J2,3 9.9 Hz, J3,4
9.1 Hz, H-3), 3.43 (t, 1H, J1,2 9.9 Hz, J2,3 9.9 Hz, H-2); 13C NMR
5.15; N, 8.04; S, 6.05.
4.8. Phenyl 2-azido-3-O-benzyl-2-deoxy-1-thio-b-
D-glucopyranoside (16)
(50 MHz, CDCl3):
d 166.2 (C(O)CH2Cl), 154.5, 152.6, 137.6, 133.9,
A solution of 15 (2.3 g, 4.4 mmol) in 60% aq AcOH (40 mL) was
130.8, 129.2, 128.8, 128.7, 128.2, 115.9, 114.9 (aromatic), 86.4 (C-1),
82.5, 76.5, 75.6, 72.3 (C-3,4,5, PhCH2), 68.2 (C-6), 65.1 (C-2), 55.8
(OCH3), 40.5 (C(O)CH2Cl). Anal. Calcd for C28H28ClN3O6S: C, 58.99; H,
4.95; N, 7.37; S, 5.62. Found: C, 58.64; H, 4.92; N, 7.35; S, 5.65.
heated under reflux for 6 h. The mixture was cooled to room
temperature and concentrated. The residue was dissolved in dry
MeOH (50 mL) and a catalytic amount of NaOMe was added. The
mixture was stirred overnight at room temperature, neutralized
with Amberlite IR120 (Hþ) resin, filtered, and concentrated. The
residue was purified by column chromatography (9:1/4:1 tolu-
4.11. Methyl [2-azido-3-O-benzyl-4-O-chloroacetyl-2-deoxy-
6-O-(4-methoxy)phenyl-
butyl 2-O-benzoyl-3-O-benzyl-a-L
a
-
D
-glucopyranosyl]-(1/4)-(tert-
-idopyranosideuronate) (19)
ene/EtOAc) to give 16 as a syrup (1.5 g, 90%); [
a
]
D ꢁ72 (c 0.3, CHCl3);
1H NMR (200 MHz, CDCl3):
d
7.56–7.15 (m, 10H, aromatic), 4.92 (d,
1H, J 11.4 Hz, PhCH2), 4.75 (d, 1H, J 11.4 Hz, PhCH2), 4.46 (m, 1H, H-
1), 3.85 (dd, 1H, J5,6a 3.3 Hz, J6a,6b 12.1 Hz, H-6a), 3.74 (dd, 1H, J5,6b
4.4 Hz, J6a,6b 12.1 Hz, H-6b), 3.54 (m, 1H, H-5), 3.38–3.24 (m, 3H, H-
2,3,4), 2.90 (s, 1H, OH), 2.44 (s, 1H, OH); 13C NMR (50 MHz, CDCl3):
A mixture of 8 (0.72 g, 1.56 mmol), 7 (1.07 g, 1.87 mmol), and 4 Å
molecular sieves (3 g) was stirred in dry CH2Cl2 (5 mL) and dry Et2O
(15 mL) at 0 ꢀC under argon for 30 min, then DMTST (2.00 g,
7.80 mmol) was added. After 1 day, the reaction was quenched with
triethylamine (3 mL). The mixture was filtered through a pad of
Celite, the filtrate was diluted with CH2Cl2 (500 mL), and washed
with 2 M aq HCl (150 mL), saturated aq NaHCO3 (150 mL), and
water (150 mL), dried, and concentrated. The residue was purified
by column chromatography (98:2/9:1 toluene/EtOAc) to give 19
d
137.9, 133.4, 131.4, 129.3, 129.2, 128.8, 128.6, 128.4, 128.3, 125.4
(aromatic), 86.6 (C-1), 84.8, 79.5, 75.6, 70.3 (C-3,4,5, PhCH2), 65.1
(C-2), 62.5 (C-6). Anal. Calcd for C19H21N3O4S: C, 58.90; H, 5.46; N,
10.85; S, 8.28. Found: C, 58.48; H, 5.45; N, 10.83; S, 8.27.
4.9. Phenyl 2-azido-3-O-benzyl-2-deoxy-6-O-(4-
as a syrup (0.67 g, 47%); [
a]
D þ7 (c 0.3, CHCl3); 1H NMR (400 MHz,
methoxy)phenyl-1-thio-
b-D
-glucopyranoside (18)
CDCl3): d 8.16–8.09 (m, 2H, aromatic), 7.60–7.12 (m, 13H, aromatic),
0
0
0
0
6.82–6.78 (m, 4H, aromatic), 5.24 (dd, 1H, J3 ,4 9.2 Hz, J4 ,5 10.2 Hz,
To a solution of 16 (1.5 g, 3.8 mmol) in dry CH2Cl2 (40 mL),
triethylamine (1.1 mL, 7.6 mmol), 4-dimethylaminopyridine
(0.02 g, 0.2 mmol), and tosyl chloride (0.9 g, 4.6 mmol) were added.
The mixture was stirred overnight at room temperature. Ice was
added, after 1 h the mixture was diluted with CH2Cl2 (300 mL),
washed with ice-cold 2 M aq HCl (100 mL), saturated aq NaHCO3
(100 mL), and water (100 mL), dried, and concentrated to give
crude 17. The solution of the residue in dry DMF (20 mL) was added
to a mixture of 4-methoxyphenol (1.2 g, 9.4 mmol) and 60% NaH
suspension (0.38 g, 9.4 mmol) in dry DMF (20 mL). The mixture was
stirred at 50 ꢀC for 3 h, cooled to room temperature, diluted with
CHCl3 (500 mL), then was washed with water (150 mL), saturated
aq NaHCO3 (150 mL), and water (150 mL), dried, and concentrated.
The residue was purified by column chromatography (98:2 toluene/
acetone) to give 18 (1.6 g, 83%); mp 103–104 ꢀC (from EtOAc/hex-
H-40), 5.15 (dd, 1H, J1,2 3.0 Hz, J2,3 3.2 Hz, H-2), 5.14 (dd, 1H, J1,2
3.0 Hz, J1,3 1.7 Hz, H-1), 4.98 (d, 1H, J1 ,2 3.6 Hz, H-10), 4.87 (d, 1H, J
11.5 Hz, PhCH2), 4.80 (d,1H, J 11.5 Hz, PhCH2), 4.68 (d,1H, J4,5 3.3 Hz,
0
0
0
0
0
0
H-5), 4.45 (d, 1H, J 11.3 Hz, PhCH2), 4.29 (ddd, 1H, J4 ,5 10.2 Hz, J5 ,6a
4.2 Hz, J5 ,6b 3.4 Hz, H-50), 4.24 (ddd, 1H, J1,3 1.7 Hz, J2,3 3.2 Hz, J3,4
0
0
4.8 Hz, H-3), 4.23 (d, 1H, J 11.3 Hz, PhCH2), 4.12 (dd, 1H, J3,4 4.8 Hz,
J4,5 3.3 Hz, H-4), 4.04 (dd, 1H, J5 ,6a 4.2 Hz, J6a ,6b 10.5 Hz, H-6a0),
0
0
0
0
3.90 (dd, 1H, J5 ,6b 3.4 Hz, J6a ,6b 10.5 Hz, H-6b0), 3.77 (dd, 1H, J2 ,3
0
0
0
0
0
0
10.0 Hz, J3 ,4 9.2 Hz, H-30), 3.77 (d, 1H, J 14.3 Hz, C(O)CH2Cl), 3.75 (s,
0
0
3H, Ar–OCH3), 3.72 (d, 1H, J 14.3 Hz, C(O)CH2Cl), 3.50 (s, 3H, OCH3),
3.35 (dd,1H, J1 ,2 3.6 Hz, J2 ,3 10.0 Hz, H-2 ),1.52 (s, 9H, C(CH3)3); 13C
0
0
0
0
0
NMR (100 MHz, CDCl3):
d 168.2 (C-6), 165.7 (OC(O)CH2Cl), 165.5
(OC(O)Ph), 154.3, 152.6, 137.6, 137.5, 133.4, 130.0, 129.8, 129.7, 128.6,
128.5, 128.4, 128.1, 127.9, 127.7, 115.9, 114.7 (aromatic), 100.3 (C-1),
98.3 (C-10), 82.8 (C(CH3)3), 77.9 (C-30), 74.7 (PhCH2), 74.4 (C-4), 74.1
(C-3), 73.3 (PhCH2), 72.5 (C-40), 70.1 (C-2), 69.1 (C-5), 68.8 (C-50),
67.4 (C-60), 63.3 (C-20), 56.2 (OCH3), 55.7 (ArOCH3), 40.4
(C(O)CH2Cl), 28.3 (C(CH3)3). Anal. Calcd for C47H52ClN3O14: C, 61.47;
H, 5.71; N, 4.58. Found: C, 61.03; H, 5.66; N, 4.49.
anes); [
a]
ꢁ40 (c 0.5, CHCl3); 1H NMR (200 MHz, CDCl3):
d 7.62–
D
6.76 (m, 14H, aromatic), 4.95 (d, 1H, J 11 Hz, PhCH2), 4.77 (d, 1H, J
11 Hz, PhCH2), 4.47 (m, 1H, H-1), 4.23 (dd, 1H, J5,6a 2.9 Hz, J6a,6b
10.3 Hz, H-6a), 4.14 (dd, 1H, J5,6b 4.8 Hz, J6a,6b 10.3 Hz, H-6b), 3.76 (s,
3H, OCH3), 3.70–3.53 (m, 2H, H-4,5), 3.42–3.29 (m, 2H, H-2,3), 2.50
(d, 1H, J2,OH 2.9 Hz, OH); 13C NMR (50 MHz, CDCl3):
d
154.3, 152.8,
4.12. Methyl [2-azido-3-O-benzyl-2-deoxy-6-O-(4-methoxy)-
137.9, 133.6, 131.4, 129.1, 128.9, 128.5, 128.4, 128.3, 115.9, 114.8 (ar-
omatic), 86.5 (C-1), 84.9, 78.0, 75.6, 70.7 (C-3,4,5, PhCH2), 68.4 (C-
6), 64.8 (C-2), 55.9 (OCH3). Anal. Calcd for C26H27N3O5S: C, 63.27; H,
5.51; N, 8.51; S, 6.50. Found: C, 63.03; H, 5.48; N, 8.54; S, 6.53.
phenyl-
3-O-benzyl-
a
-
D
-glucopyranosyl]-(1/4)-(tert-butyl 2-O-benzoyl-
-idopyranosideuronate) (20)
a-L
To a solution of 19 (0.67 g, 0.74 mmol) in dry DMF (7 mL), a so-
lution of hydrazinedithiocarbonate54 (2.21 mmol) in 5.3 mL of
ethanol/water/dioxane (4:2:1) was added. The mixture was stirred
at room temperature for 1 h, diluted with CH2Cl2 (250 mL), washed
with 2 M aq HCl (50 mL), saturated aq NaHCO3 (50 mL), and water
(50 mL), dried, and concentrated. The residue was purified by col-
umn chromatography (9:1 toluene/EtOAc) to give 20 as a syrup
4.10. Phenyl 2-azido-3-O-benzyl-4-O-chloroacetyl-2-deoxy-6-
O-(4-methoxy)phenyl-1-thio-b-D-glucopyranoside (7)
A solution of 90% chloroacetic anhydride (0.90 g, 0.75 mmol) in
CH2Cl2 (12 mL) was added dropwise to a stirred solution of 18
(1.6 g, 3.2 mmol) in dry CH2Cl2 (46 mL) and dry pyridine (12 mL) at
ꢁ20 ꢀC. After 20 min, the reaction was quenched with water
(10 mL) and stirred for 20 min. The mixture was diluted with
CH2Cl2 (200 mL), washed with 2 M aq HCl (75 mL), saturated aq
(0.49 g, 80%); [
a
]
D
þ2 (c 0.9, CHCl3); 1H NMR (200 MHz, CDCl3):
d
8.15–8.10 (m, 2H, aromatic), 7.58–7.10 (m, 13H, aromatic), 6.89–
6.77 (m, 4H, aromatic), 5.15 (dd, 1H, J1,2 2.7 Hz, J2,3 3.2 Hz, H-2), 5.14
(d, 1H, J1,2 2.7 Hz, H-1), 4.96 (d, 1H, J1 ,2 3.3 Hz, H-10), 4.89 (d, 1H, J
0
0

J. Tatai, P. Fu¨gedi / Tetrahedron 64 (2008) 9865–9873
9871
11.7 Hz, PhCH2), 4.78 (d,1H, J 11.7 Hz, PhCH2), 4.65 (d,1H, J4,5 3.3 Hz,
H-5), 4.46 (d, 1H, J 11.0 Hz, PhCH2), 4.34 (d, 1H, J 11.0 Hz, PhCH2),
(0.31 g, 93%); [
a
]
ꢁ32 (c 0.4, CHCl3); 1H NMR (200 MHz, CDCl3):
D
d
7.40–7.14 (m, 22H, aromatic), 6.89–6.75 (m, 2H, aromatic), 5.26 (d,
4.25–4.08 (m, 5H, H-3,4,30,50,6a0), 3.83 (dd, 1H, J5 ,6b 3.3 Hz, J6a ,6b
1H, J1 ,2 2.6 Hz, H-100), 5.06 (d, 1H, J1 ,2 3.7 Hz, H-10), 5.00 (d, 1H, J
10.3 Hz, PhCH2), 4.94 (s, 1H, H-1), 4.74 (d, 1H, J 12.1 Hz, PhCH2), 4.73
(d, 1H, J 12.0 Hz, PhCH2), 4.62 (d, 1H, J4,5 2.9 Hz, H-5), 4.60 (d, 1H, J
11.7 Hz, PhCH2), 4.59 (d, 1H, J 10.6 Hz, PhCH2), 4.57 (d, 1H, J 12.0 Hz,
0
0
0
0
00 00
0
0
8.8 Hz, H-6b0), 3.75 (s, 3H, ArOCH3), 3.62 (ddd, 1H, J3 ,4 9.9 Hz, J4 ,5
0
0
0
0
9.9 Hz, J4 ,OH 3.30Hz, H-40), 3.49 (s, 3H, OCH3), 3.20 (dd,1H, J1 ,2 3.3 Hz,
0
0
0
0
0
0
J2 ,3 9.9 Hz, H-2 ), 2.41 (d,1H, J4 ,OH 3.3 Hz, OH),1.49 (s, 9H, C(CH3)3);
13C NMR (50 MHz, CDCl3):
d
168.2 (C-6), 165.6 (OC(O)Ph), 154.2,
PhCH2), 4.57 (d, 1H, J4 ,5 1.8 Hz, H-5 ), 4.53 (d, 1H, J 12.4 Hz, PhCH2),
00
00 00
152.9, 138.1, 137.7, 133.4, 130.1, 129.8, 129.1, 128.7, 128.6, 128.5, 128.3,
128.1,128.0,127.9,125.4,115.8,114.7 (aromatic),100.3 (C-1), 98.9 (C-
10), 82.8 (C(CH3)3), 80.0 (C-30), 74.8, 74.52, 74.47 (C-3,4, PhCH2), 73.3
(PhCH2), 71.3, 70.6, 70.1, 69.2(C-2,5,40,50), 67.8(C-60), 63.2(C-20), 56.3
4.47 (d,1H, J 11.7 Hz, PhCH2), 4.25–4.16 (m, 2H, H-3,40), 4.15–4.06 (m,
3H,H-50,6a0,6b0), 4.01(t,1H,J3 ,4 1.8 Hz,J4 ,5 1.8 Hz,H-4 ), 3.91(t,1H,
00
00 00
00 00
J3,4 2.9 Hz, J4,5 2.9 Hz, H-4), 3.80 (m, 1H, H-300), 3.75 (s, 3H, ArOCH3),
3.73–3.68 (m, 2H, H-2,30), 3.61 (dd,1H, J1 ,2 3.7 Hz, J2 ,3 10.3 Hz, H-2 ),
0
0
0
0
0
(OCH3), 55.8 (ArOCH3), 28.3 (C(CH3)3). Anal. Calcd for C45H51N3O13
:
3.51 (m,1H, H-200), 3.48 (s, 3H, OCH3),1.53 (s, 9H, C(CH3)3),1.32 (s, 9H,
C, 64.20; H, 6.11; N, 4.99. Found: C, 63.81; H, 6.03; N, 4.86.
C(CH3)3); 13C NMR (50 MHz, CDCl3): 169.0, 168.6 (C-6, C-600), 154.2,
d
152.7, 138.0, 137.70, 137.66, 137.0, 129.1, 128.7, 128.62, 128.57, 128.3,
128.22, 128.18, 128.14, 128.09, 128.0, 127.6, 125.4, 116.1, 114.7 (aro-
matic), 102.9, 100.9, 95.3 (C-1,10,100), 82.6 (C(CH3)3), 82.0 (C(CH3)3),
80.0 (C-30), 75.8, 75.5, 74.71, 74.70, 73.4, 73.0, 72.8, 72.5, 71.4, 70.8,
69.3, 68.6, 68.0, 67.7, 66.6 (C-60), 64.0 (C-20), 56.2, 55.8 (OCH3,
4.13. Methyl (tert-butyl 2-O-benzoyl-3,4-di-O-benzyl-
idopyranosyluronate)-(1/4)-[2-azido-3-O-benzyl-2-deoxy-6-
O-(4-methoxy)phenyl- -glucopyranosyl]-(1/4)-(tert-butyl
2-O-benzoyl-3-O-benzyl- -idopyranosideuronate) (5)
a-L-
a-D
a-L
ArOCH3), 28.3 (C(CH3)3), 28.1 (C(CH3)3). Anal. Calcd for C62H75N3O18
:
A mixture of 6 (0.45 g, 0.72 mmol), 20 (0.40 g, 0.48 mmol), and
4 Å molecular sieves (1.0 g) was stirred in dry CH2Cl2 (10 mL) at 0 ꢀC
under argon for 30 min, then DMTST (0.62 g, 2.40 mmol) was
added. After 3 days, the reaction was quenched with triethylamine
(1 mL). The mixture was filtered through a pad of Celite, the filtrate
was diluted with CH2Cl2 (250 mL) and washed with 2 M aq HCl
(50 mL), saturated aq NaHCO3 (50 mL), and water (50 mL), dried,
and concentrated. The residue was purified by column chroma-
tography (95:5/9:1 toluene/EtOAc) to give 5 as a syrup (0.40 g,
C, 64.74; H, 6.57; N, 3.65. Found: C, 64.38; H, 6.52; N, 3.59.
4.15. Methyl (tert-butyl 3,4-di-O-benzyl-a-L-idopyrano-
syluronate)-(1/4)-(2-azido-3-O-benzyl-2-deoxy-
a
-
-
D
-glucopyranosyl)-(1/4)-(tert-butyl 3-O-benzyl-
a
L-idopyranoside-uronate) (22)
To a solution of 21 (0.27 g, 0.24 mmol) in MeCN (4.5 mL) and
water (0.5 mL), ceric ammonium nitrate (0.39 g, 0.72 mmol) was
added. The mixture was stirred at room temperature for 1 h, neu-
tralized with saturated aq NaHCO3 (2 mL), diluted with CHCl3
(300 mL), washed with saturated aq NaHCO3 (50 mL) and water
(50 mL), dried, and concentrated. The residue was purified by col-
umn chromatography (9:1/4:1 toluene/acetone) to give 22
61%); [
a]
D þ3 (c 0.3, CHCl3); 1H NMR (400 MHz, CDCl3):
d 8.14–8.05
(m, 2H, aromatic), 7.86–7.78 (m, 2H, aromatic), 7.59–7.42 (m, 4H,
aromatic), 7.37–7.12 (m, 22H, aromatic), 6.91–6.82 (m, 2H, aro-
matic), 6.77–6.68 (m, 2H, aromatic), 5.55 (d, 1H, J1 ,2 5.6 Hz, H-100),
00 00
00 00
00 00
00 00
00
5.14 (ddd, 1H, J1 ,2 5.6 Hz, J2 ,3 4.4 Hz, J2 ,4 0.7 Hz, H-2 ), 5.12 (ddd,
1H, J1,2 3.3 Hz, J2,3 4.2 Hz, J2,4 0.5 Hz, H-2), 5.09 (d, 1H, J1,2 3.3 Hz, H-
(0.21 g, 84%) as a syrup; [
a
]
D ꢁ40 (c 0.3, CHCl3); 1H NMR (400 MHz,
1), 4.90 (d, 1H, J1 ,2 3.7 Hz, H-10), 4.84 (d, 1H, J 11.5 Hz, PhCH2), 4.77
(d, 1H, J 11.5 Hz, PhCH2), 4.74 (d, 1H, J 11.1 Hz, PhCH2), 4.73 (d, 1H, J
11.7 Hz, PhCH2), 4.62 (d,1H, J 11.7 Hz, PhCH2), 4.57 (d,1H, J4,5 3.6 Hz,
CDCl3):
d
7.41–7.15 (m, 20H, aromatic), 5.30 (d, 1H, J1 ,2 3.5 Hz, H-
0
0
00 00
100), 5.00 (d, 1H, J1 ,2 3.7 Hz, H-1 ), 4.94 (d, 1H, J 11.6 Hz, PhCH2), 4.92
(d, 1H, J1,2 1.8 Hz, H-1), 4.75 (d, 1H, J 11.5 Hz, PhCH2), 4.67 (d, 1H, J4,5
2.4 Hz, H-5), 4.65 (s, 1H, PhCH2), 4.63 (s, 1H, PhCH2), 4.62 (d, 1H, J
0
0
0
H-5), 4.50 (d, 1H, J 11.5 Hz, PhCH2), 4.48 (d, 1H, J4 ,5 5.1 Hz, H-500),
00 00
00
0
0
0
0
00 00
4.42 (d,1H, J 11.5 Hz, PhCH2), 4.22 (dd,1H, J3 ,4 9.0 Hz, J4 ,5 9.8 Hz H-
40), 4.18 (dd, 1H, J2,3 4.2 Hz, J3,4 4.8 Hz, H-3), 4.12 (d, 1H, J 11.1 Hz,
PhCH2), 4.1–4.05 (m, 3H, H-50,6a0,6b0), 4.03 (ddd, 1H, J2,4 0.5 Hz, J3,4
11.5 Hz, PhCH2), 4.60 (d,1H, J4 ,5 2.5 Hz, H-5 ), 4.58 (d,1H, J 11.6 Hz,
PhCH2), 4.57 (d, 1H, J 11.6 Hz, PhCH2), 4.54 (d, 1H, J 11.6 Hz, PhCH2),
4.14 (dd, 1H, J3,4 3.8 Hz, J4,5 2.4 Hz, H-4), 3.90 (dd, 1H,0J2,3 3.2 Hz, J3,4
00 00
00 00
00 00
0 0 0 0
3.8 Hz, H-3), 3.89 (dd,1H, J3 ,4 8.8 Hz, J4 ,5 9.8 Hz, H-4 ), 3.84 (m, 2H,
4.8 Hz, J4,5 3.6 Hz, H-4), 3.88 (ddd, 1H, J2 ,4 0.7 Hz, J3 ,4 5.6 Hz, J4 ,5
5.1 Hz, H-400), 3.82 (dd, 1H, J2 ,3 4.4 Hz, J3 ,4 5.6 Hz, H-300), 3.73 (s,
H-6a0,6b0), 3.81 (dd, 1H, J3 ,4 5.0 Hz, J4 ,5 2.5 Hz, H-400), 3.80 (dd,
00 00
00 00
00 00 00 00
3H, Ar–OCH3), 3.64 (dd, 1H, J2 ,3 10.2 Hz, J3 ,4 9.0 Hz, H-30), 3.45 (s,
1H, J2 ,3 5.4 Hz, J3 ,4 5.0 Hz, H-300), 3.78 (m, 1H, H-50), 3.72 (dd, 1H,
00 00 00 00
0
0
0
0
3H, OCH3), 3.30 (dd, 1H, J1 ,2 3.7 Hz, J2 ,3 10.2 Hz, H-20), 1.41 (s, 9H,
J2 ,3 10.0 Hz, J3 ,4 8.8 Hz, H-30), 3.71 (ddd, 1H, J1,2 1.8 Hz, J2,3 3.2 Hz,
0
0
0
0
0
0
0
0
C(CH3)3), 1.39 (s, 9H, C(CH3)3); 13C NMR (100 MHz, CDCl3):
d
168.6
J2,OH 10.8 Hz, H-2), 3.58 (ddd, 1H, J1 ,2 3.5 Hz, J2 ,3 5.4 Hz, J2 ,OH 7.0
00 00 00 00 00
(C-600), 168.2 (C-6), 165.5 (OC(O)Ph), 165.3 (OC(O)Ph), 154.0, 152.7,
138.4, 137.7, 137.4, 133.4, 133.2, 129.99, 129.96, 129.6, 129.5, 128.7,
128.4,128.3,128.1,128.04,128.00,127.9,127.82,127.77,127.75,127.7,
127.3, 116.3, 114.4 (aromatic), 100.1 (C-1), 98.8 (C-10), 98.0 (C-100),
82.7 (C(CH3)3), 81.8 (C(CH3)3), 78.4 (C-30), 76.1 (C-400), 76.0 (C-40),
75.8 (C-300), 74.8 (C-4), 74.7 (PhCH2), 74.5 (C-3), 73.3 (PhCH2), 73.2
(PhCH2), 73.1 (PhCH2), 72.3 (C-200), 72.0 (C-500), 70.8 (C-50), 69.9 (C-
2), 69.2 (C-5), 66.4 (C-60), 63.4 (C-20), 56.2 (OCH3), 55.7 (ArOCH3),
28.2 (C(CH3)3), 28.1 (C(CH3)3). Anal. Calcd for C76H83N3O20: C, 67.19;
H, 6.16; N, 3.09. Found: C, 66.98; H, 6.19; N 3.02.
Hz, H-200), 3.51 (dd, 1H, J1 ,2 3.7 Hz, J2 ,3 10.0 Hz, H-20), 3.47 (s, 3H,
0
0
0
0
00
OCH3), 3.42 (d, 1H, J2,OH 10.8 Hz, C-2-OH), 3.08 (d, 1H, J2 ,OH 7.0 Hz,
C-200-OH), 2.20 (br s, 1H, C-60-OH), 1.52 (s, 9H, C(CH3)3), 1.34 (s, 9H,
C(CH3)3); 13C NMR (100 MHz, CDCl3):
d
169.1 (C-6), 168.8 (C-600),
138.2, 137.81, 137.77, 137.2, 128.83, 128.76, 128.7, 128.44, 128.40,
128.37, 128.33, 128.25, 128.2, 128.1, 127.7 (aromatic), 103.1 (C-1),
100.8 (C-100), 95.5 (C-10), 83.0 (C(CH3)3), 82.4 (C(CH3)3), 79.8 (C-30),
76.3 (C-400), 75.6 (PhCH2), 75.3 (C-300), 75.0 (C-40), 73.5 (PhCH2), 73.4
(PhCH2), 73.1 (C-3), 72.7 (PhCH2), 72.4 (C-50), 71.7 (C-4), 69.9 (C-500),
69.4 (C-200), 68.0 (C-5), 67.8 (C-2), 64.2 (C-20), 61.6 (C-60), 56.3
(OCH3), 28.4 (C(CH3)3), 28.2 (C(CH3)3). Anal. Calcd for C55H69N3O17
:
4.14. Methyl (tert-butyl 3,4-di-O-benzyl-
syluronate)-(1/4)-[2-azido-3-O-benzyl-2-deoxy-6-O-
(4-methoxy)phenyl- -glucopyranosyl]-(1/4)-(tert-butyl
3-O-benzyl- -idopyranosideuronate) (21)
a-L-idopyrano-
C, 63.27; H, 6.66; N, 4.02. Found: C, 62.91; H, 6.62; N, 3.96.
a
-D
4.16. Methyl (3,4-di-O-benzyl-
(1/4)-(2-azido-3-O-benzyl-2-deoxy-
(1/4)-(3-O-benzyl- -idopyranosideuronic acid) (23)
a
-L
-idopyranosyluronic acid)-
a
-L
a-D
-glucopyranosyl)-
a-L
To a solution of 5 (0.39 g, 0.29 mmol) in dry MeOH (5 mL),
a catalytic amount of NaOMe was added. The mixture was stirred
for one week at room temperature, neutralized with Amberlite
IR120 (Hþ) resin, filtered, and concentrated to give 21 as a syrup
A solution of compound 22 (0.19 g, 0.19 mmol) in a 20% solution
of CF3CO2H in CH2Cl2 (6 mL) was stirred for 1 h, the mixture was
concentrated, and the residue was co-evaporated with toluene

9872
J. Tatai, P. Fu¨gedi / Tetrahedron 64 (2008) 9865–9873
(50 mL) to give 23 (0.17 g, 98%) as a syrup; [
a
]
D ꢁ30 (c 0.3, CHCl3);
CHCl3); 1H NMR (200 MHz, CDCl3):
d 7.39–7.11 (m, 20H, aromatic),
1H NMR (400 MHz,00CDCl3): 7.40–7.10 (m, 20H, aromatic), 5.22 (d,
6.89–6.72 (m, 4H, aromatic), 5.16, 4.96 (2s, 2H, H-1,100), 4.91 (d, 1H,
1H, J1 ,2 2.0 Hz, H-1 ), 4.94 (s, 1H, H-1), 4.83 (br s, 1H, H-5), 4.77 (d,
J1 ,2 3.7 Hz, H-10), 4.87 (d, 1H, J4,5 1.1 Hz, H-5), 4.71 (d, 2H, J 11.7 Hz,
00 00
0
0
1H, J1 ,2 4.1 Hz, H-10), 4.71 (d, 1H, J 11.5 Hz, PhCH2), 4.70 (d, 1H, J
11.5 Hz, PhCH2), 4.65 (br s, 1H, H-500), 4.62 (d, 1H, J 11.7 Hz, PhCH2),
4.56 (d, 1H, J 11.6 Hz, PhCH2), 4.55 (d, 1H, J 11.6 Hz, PhCH2), 4.53 (d,
1H, J 11.5 Hz, PhCH2), 4.51 (s, 2H, PhCH2), 4.14 (s,1H, H-4), 3.89–3.67
PhCH2), 4.71 (d, 1H, J4 ,5 2.6 Hz, H-500), 4.65 (d, 1H, J 12.1 Hz,
0
0
00 00
PhCH2), 4.59 (d, 1H, J 11.0 Hz, PhCH2), 4.55 (d, 1H, J 12.1 Hz, PhCH2),
4.54 (d, 1H, J 11.4 Hz, PhCH2), 4.45 (d, 1H, J 11.7 Hz, PhCH2), 4.44 (d,
1H, J 11.4 Hz, PhCH2), 4.23–4.03 (m, 4H), 3.96–3.71 (m, 5H), 3.68 (s,
(m, 10H), 3.49 (dd, 1H, J1 ,2 4.1 Hz, J2 ,3 10.1 Hz, H-20); 13C NMR
3H, ArOCH3), 3.68–3.63 (m, 2H), 3.59 (dd, 1H, J1 ,2 3.7 Hz, J2 ,3
0
0
0
0
0
0
0
0
(100 MHz, CDCl3):
d
171.4, 171.1 (C-6,600), 137.44, 137.37, 137.2, 136.5,
9.9 Hz, H-20), 3.43 (s, 3H, OCH3); 13C NMR (CDCl3):
d
171.7, 170.9 (C-
128.6, 128.41, 128.37, 128.3, 128.19, 128.15, 127.9, 127.67, 127.65
(aromatic), 103.2 (C-1), 99.9 (C-100), 95.7 (C-10), 78.9, 75.41, 75.40
(PhCH2), 75.2, 74.6, 73.9 (PhCH2), 73.0 (PhCH2), 72.9, 72.2, 71.8, 71.9
(PhCH2), 68.9, 68.8, 66.2, 66.0, 64.0, 61.4 (C-60), 56.3 (OCH3).
6,600), 154.4, 152.5, 138.0, 137.5, 137.4, 137.1, 136.4, 129.1, 128.7, 128.5,
128.4, 128.3, 128.2, 127.9, 127.8, 127.5, 125.4, 116.1, 114.8 (aromatic),
103.4 (C-1), 100.6 (C-100), 94.9 (C-1), 79.1 (C-30), 75.3, 75.2, 74.9,
73.9, 73.7, 72.6, 72.1, 71.3, 71.1 (2C), 68.5, 67.8, 66.7, 66.5, 66.0 (C-60),
64.1 (C-20), 56.5, 55.7 (ArOCH3, OCH3).
4.17. Methyl (2-O-sulfonato-
(1/4)-(2-deoxy-2-sulfonatamido-6-O-sulfonato-
glucopyranosyl)-(1/4)-(2-O-sulfonato-
idopyranosideuronic acid) hexasodium salt (3)
a-L-idopyranosyluronic acid)-
a
-D-
4.19. Methyl (2-O-sulfonato-
(1/4)-(2-deoxy-2-sulfonatamido-a-D
a
-
L
-idopyranosyluronic acid)-
-glucopyranosyl)-
(1/4)-(2-O-sulfonato-a-L-idopyranosideuronic acid)
a
-L
-
pentasodium salt (4)
To a solution of 23 (0.15 g, 0.16 mmol) in dry DMF (3.0 mL),
sulfur trioxide pyridine complex (0.15 g, 0.96 mmol) was added.
The mixture was stirred at room temperature for 3 h, neutralized
with saturated aq NaHCO3 (1 mL), and concentrated. The residue
was purified by column chromatography (4:1/2:1 CH2Cl2/MeOH),
and the product was eluted from a column of Dovex-50W X8 resin
using MeOH to give 24 (0.14 g, 68%) as a foam; 1H NMR (200 MHz,
Compound 26 (0.18 g, 0.17 mmol) was O-sulfated as described
for compound 24. The product was purified by column chroma-
tography (12:2:1 EtOAc/MeOH/H2O), then it was eluted from a col-
umn of Dovex-50W X8 resin using MeOH to give 27 (0.22 g, 99%) as
a foam; 1H NMR (200 MHz, CD3OD):
d 7.42–7.07 (m, 20H, aromatic),
6.91–6.75 (m, 4H, aromatic), 5.90, 5.28, 5.07 (3s, 3H, H-1,10,100), 5.10
(d, 1H, J 11.0 Hz, PhCH2), 5.00 (d, 1H, J 11.0 Hz, PhCH2), 4.80 (s, 1H),
4.79 (d,1H, J 11.7 Hz, PhCH2), 4.77 (s,1H), 4.70–4.18 (m, 11H), 4.11 (s,
CD3OD):
5.10–3.70 (m, 23H), 3.53 (dd,1H, J1 ,2 3.7 Hz, J2 ,3 10.1 Hz, H-2 ), 3.39
(s, 3H, OCH3); 13C NMR (50 MHz, CD3OD): 177.0, 176.9 (C-6,600),
d
7.41–7.08 (m, 20H, aromatic), 5.84, 5.33 (2s, 2H, H-1,100),
0
0
0
0
0
0 0
d
1H), 4.03 (s, 2H), 3.88 (s, 2H), 3.70 (s, 3H, ArOCH3), 3.64 (dd, 1H, J1 ,2
2.9 Hz, J2 ,3 9.2 Hz, H-20), 3.39 (s, 3H, OCH3); 13C NMR (50 MHz,
CD3OD):
175.9, 175.2 (C-6,600), 153.8, 153.0, 138.1, 137.03, 136.96,
0
0
140.0, 139.0, 138.9, 138.5, 130.1, 129.4, 129.20, 129.16, 129.0, 128.9,
128.8, 128.6, 128.4 (aromatic), 101.6, 96.0, 94.2 (C-1,10,100), 77.0, 75.9,
75.0, 73.8, 72.8, 72.6, 71.8, 70.97, 70.95, 70.9, 70.2, 70.1, 69.3, 68.3,
66.1, 56.4 (OCH3); ESIMS (ꢁ): m/z 1235.6 [Mꢁ2NaþH]ꢁ.
d
136.1, 128.5, 128.1, 128.0, 127.9, 127.6, 127.4, 127.3, 127.1, 115.6, 114.5
(aromatic), 99.8, 94.0, 93.0 (C-1,10,100), 77.2, 75.8, 75.5, 73.8, 72.4, 71.7
(2C), 71.0 (2C), 70.3, 69.8, 69.4, 69.4, 68.9, 67.7, 66.9, 64.2, 55.2 (2C,
ArOCH3, OCH3); ESIMS (ꢁ): m/z 1253.9 [Mꢁ3Naþ2NH4]ꢁ.
A solution of 24 (0.14 g, 0.11 mmol) in THF (2.5 mL) and water
(2.5 mL) was hydrogenated with 10% Pd/C (0.05 g) under atmo-
spheric pressure at room temperature. After 2 days, the mixture
was filtered through a pad of Celite and concentrated to give 25. No
aromatic signal was visible in the NMR spectra.
To a solution of 27 (0.22 g, 0.17 mmol) in MeCN (4.5 mL) and
water (0.5 mL), ceric ammonium nitrate (0.27 g, 0.50 mmol) was
added. The mixture was stirred at room temperature for 3 h, neu-
tralized with saturated aq NaHCO3 (2 mL), and concentrated. The
residue was purified by column chromatography (12:2:1 EtOAc/
MeOH/H2O), and the product was eluted from a column of Dovex-
50W X8 resin using MeOH to give 28 (0.16 g, 83%) as a foam; 1H
The pH of a solution of 25 in water (3 mL) was adjusted to 9.8
with 1 M aq NaOH, then sulfur trioxide trimethylamine complex
(0.092 g, 0.66 mmol) was added in six portions hourly, the pH being
maintained at 9.8 by the addition of 0.5 M aq NaOH. After 6 h, the
mixture was neutralized with 1 M aq HCl and concentrated. The
residue was purified by column chromatography (2:2:1 EtOAc/
MeOH/H2O) to give trisaccharide fractions, which were eluted from
a column of Dovex-50W X8 resin using H2O, and desalted on
a column of Sephadex G-25 using water as eluent to afford 3
NMR (200 MHz, CD3OD):
d 7.40–7.11 (m, 20H, aromatic), 5.82, 5.19
(2s, 2H, H-1,100), 5.08 (d, 1H, J 11.0 Hz, PhCH2), 5.01–4.86 (m, 2H),
0
0
0
0
4.82–3.90 (m, 16H), 3.78–3.51 (m, 3H), 3.46 (dd, 1H, J1 ,2 3.3 Hz, J2 ,3
9.9 Hz, H-20), 3.38 (s, 3H, OCH3); 13C NMR (50 MHz, CD3OD):
d
177.6,
176.9 (C-6,600), 140.0, 139.1, 138.8, 138.3, 130.0, 129.5, 129.4, 129.3,
129.2, 128.92, 128.87, 128.7, 128.4 (aromatic), 101.7, 95.9, 94.7 (C-
1,10,100), 78.2, 77.6, 75.70, 75.69, 73.61, 73.60, 72.69 (2C), 72.68 (2C),
71.89, 71.87, 70.75, 70.73, 69.1, 65.9, 63.1, 56.3 (OCH3); ESIMS (ꢁ):
m/z 1112.7 [Mꢁ3Naþ2H]ꢁ.
(0.049 g, 45%) as a foam; 1H NMR (400 MHz, D2O):
d 5.23 (d,1H, J1 ,2
0 0
3.3 Hz, H-10), 5.19 (s, 1H, H-100), 5.00 (s, 1H, H-1), 4.88 (s, 1H, H-500),
4.50 (d,1H, J4,5 2.5 Hz, H-5), 4.25–4.14 (m, 5H, H-2,3,6a0,6b0,200), 4.04
(t, 1H, J2 ,3 3.2 Hz, J3 ,4 3.2 Hz, H-300), 3.98 (t, 1H, J3,4 2.5 Hz, J4,5
00 00
00 00
2.5 Hz, H-4), 3.96–3.89 (m, 2H, H-50,400), 3.74–3.64 (m, 2H, H-30,40),
Compound 28 (0.15 g, 0.13 mmol) was hydrogenated, and N-
sulfated as described for compound 3, to give 4 (0.084 g, 68%) as
3.34 (s, 3H, OCH3), 3.19 (dd, 1H, J1 ,2 3.3 Hz, J2 ,3 9.8 Hz, H-20); 13C
0
0
0
0
NMR (100 MHz, D2O):
d
175.8 (C-600), 174.4 (C-6), 99.9 (C-1), 99.4 (C-
a foam; [
a
]
D þ3 (c 0.3, H2O); 1H NMR (400 MHz, D2O):
d
5.23 (d, 1H,
100), 97.9 (C-10), 76.8 (C-40), 76.3 (C-4), 74.8 (C-2), 74.3 (C-200), 69.8
(C-30), 69.4 (C-50), 69.2 (C-400), 69.2 (C-500), 68.9 (C-300), 67.7 (C-3),
67.5 (C-5), 66.5 (C-60), 58.2 (C-20), 55.8 (OCH3); HRMS (ESI) calcd for
J1 ,2 3.5 Hz, H-10), 5.07 (dd, 1H, J1 ,2 1.4 Hz, J1 ,3 1.0 Hz, H-1 ), 4.96
00
0
0
00 00
00 00
(dd, 1H, J1,2 2.0 Hz, J1,3 3.0 Hz, H-1), 4.76 (d, 1H, J4 ,5 2.2 Hz, H-500),
00 00
00 00
00 00
4.36 (d, 1H, J4,5 2.5 Hz, H-5), 4.21 (ddd, 1H, J1 ,2 1.4 Hz, J2 ,3 3.0 Hz,
00 00
19H25NO29S4Na6 [Mꢁ6Naþ4H]2ꢁ 431.4860. Found: 431.4871.
J2 ,4 1.0 Hz, H-2 ), 4.15 (ddd, 1H, J1,3 3.0 Hz, J2,3 3.0 Hz, J3,4 2.5 Hz, H-
3), 4.14 (ddd,1H, J1,2 2.0 Hz, J2,3 3.0 Hz, J2,4 1.5 Hz, H-2), 3.99 (ddd,1H,
00
C
00
00 00 00 00 00 00
J1 ,3 1.0 Hz, J2 ,3 3.0 Hz, J3 ,4 3.4 Hz, H-3 ), 3.94 (ddd, 1H, J2,4 1.5 Hz,
4.18. Methyl (3,4-di-O-benzyl-
(1/4)-[2-azido-3-O-benzyl-2-deoxy-6-O-(4-methoxy)-
phenyl- -glucopyranosyl]-(1/4)-(3-O-benzyl-
a
-L-idopyranosyluronic acid)-
00 00
00 00
J3,4 2.5 Hz, J4,5 2.5 Hz, H-4), 3.90 (ddd, 1H, J2 ,4 1.0 Hz, J3 ,4 3.4 Hz,
a-
D
J4 ,5 2.2 Hz, H-400), 3.79 (m, 1H, H-50), 3.78 (dd, 1H, J5 ,6a 2.2 Hz,
00 00 0 0
J6a ,6b 12.4 Hz, H-6a0), 3.73 (dd, 1H, J5 ,6b 3.6 Hz, J6a ,6b 12.4 Hz, H-
0
0
0
0
0
0
a
-L-idopyranosideuronic acid) (26)
6b0), 3.63 (dd, 1H, J2 ,3 10.0 Hz, J3 ,4 8.8 Hz, H-30), 3.60 (dd, 1H, J3 ,4
0
0
0
0
0
0
0
0 0
8.8 Hz, J4 ,5 9.7 Hz, H-4 ), 3.33 (s, 3H, OCH3), 3.15 (dd, 1H, J1 ,2 3.5 Hz,
0 0
Compound 21 (0.22 g, 0.19 mmol) was hydrolyzed as described
for compound 23 to give 26 (0.18 g, 92%) as a syrup; [
a
]D ꢁ29 (c 0.2,
J2 ,3 10.0 Hz, H-2 ); 13C NMR (100 MHz, D2O): 176.7 (C-600),175.2 (C-6),
d
0
0 0

J. Tatai, P. Fu¨gedi / Tetrahedron 64 (2008) 9865–9873
9873
99.9 (C-1), 99.5 (C-100), 97.4 (C-10), 78.0 (C-40), 76.1 (C-4), 75.3 (C-2),
73.8 (C-200), 71.3 (C-50), 69.7 (C-30), 69.0 (2C, C-300,400), 68.6 (C-500),
68.2 (C-3), 68.1 (C-5), 60.0 (C-60), 58.6 (C-20), 55.6 (OCH3); HRMS
(ESI) calcd for C19H26NO26S3Na5 [Mꢁ5Naþ3H]2ꢁ 391.5075. Found:
391.5090.
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Acknowledgements
Financial support of the elemental analyzer for grant GVOP-
3.2.1-2004-04-0311/3.0 is gratefully acknowledged.
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