Synthesis of poly-O-sulfated glycosides of 2,5-anhydro-D-mannitol

Article abstract of DOI:10.1016/j.carres.2005.05.006

2,5-Anhydro-3-O-β-d-glucopyranosyl-; -3-O-α-l-idopyranosyl-; -3-O-α-d-arabinopyranosyl-; -3-O-α-l-arabinopyranosyl-; -3-O-β-d-maltopyranosyl-; -3-O-β-d-gentiobiopyranosyl-; -1,6-di-O-β-d-glucopyranosyl-; -1,6-di-O-α-l-idopyranosyl-; -1-O-β-d-maltopyranosy

Full text of DOI:10.1016/j.carres.2005.05.006

Carbohydrate
RESEARCH
Carbohydrate Research 340 (2005) 1739–1749
Note
Synthesis of poly-O-sulfated glycosides of 2,5-anhydro-D-mannitolq
*
Janos Kuszmann, Gabor Medgyes and Sandor Boros
´
´
´
IVAX Drug Research Institute, H-1325 Budapest, PO Box 82, Hungary
Received 18 April 2005; accepted 18 May 2005
´ ´
Dedicated to Andras Liptak on the occasion of his 70th birthday
Abstract—2,5-Anhydro-3-O-b-D-glucopyranosyl-; -3-O-a-L-idopyranosyl-; -3-O-a-D-arabinopyranosyl-; -3-O-a-L-arabinopyranos-
yl-; -3-O-b-D-maltopyranosyl-; -3-O-b-D-gentiobiopyranosyl-; -1,6-di-O-b-D-glucopyranosyl-; -1,6-di-O-a-L-idopyranosyl-; -1-O-b-
D-maltopyranosyl-; -1,3,6-tri-O-b-D-glucopyranosyl-; -1,6-di-O-b-maltopyranosyl- and -1,6-di-O-b-D-gentiobiopyranosyl-2,5-anhy-
dro-D-mannitol as well as their poly-O-sulfated derivatives were synthesized. The IP3–IC50 values of their sodium and/or potassium
salts were determined for structure–activity studies aiming at the synthesis of new, orally active antiasthmatic compounds.
ꢀ 2005 Elsevier Ltd. All rights reserved.
Keywords: 2,5-Anhydro-D-mannitol derivatives; Mono-, di-, tri-, tetra- and pentasaccharides; Poly-O-sulfated saccharides; Antiasthmatic compounds
Heparin, well known as an anticoagulant,1 possesses
other very important and significant biological activities
too, among others antiinflammatory, antiallergic and
even antiasthmatic properties.2–5 These activities are
however independent from the anticoagulant activity
of native heparin, and are displayed by low, and even
ultralow molecular weight heparin fragments, obtained
by degradation of the original GAGs.6,7 Recently, it
was claimed by Ahmed and Smith8 that a mixture of
poly-O-sulfated disaccharides, obtained by degradation
of heparin with HNO2 and subsequent reduction with
NaBH4 followed by O-sulfation, possesses pronounced
antiasthmatic activity. As the separation of the two
main components of this mixture, that is, the hepta
sodium salt of 2,5-anhydro-3-O-(b-D-glucopyranosylur-
onate)- and (a-L-idopyranosyluronate)-D-mannitol
hexa-O-sulfate 1 and 2 (Fig. 1), represented a very diffi-
cult problem, both were synthesized9 and differed signif-
icantly in their biological activity. For establishing the
structure–activity relationship of this type of polysulf-
ated glycosides, further analogues were synthesized
and tested for their biological activity. These com-
pounds inhibit the binding of inositol-1,4,5-triphosphate
(IP3) to its receptor in microsomal membrane pre-
parations. As IP3 is a messenger molecule playing
distinguished role in the activation of different cells,
interfering with this function can explain the antiasth-
matic effect of these glycosides. The IP3 antagonist effect
was determined using rat cerebellum membrane prepa-
rations,10 and the obtained IC50 data are given in paren-
thesis for each polysulfated new compound.
First, we decided to investigate the role of the carbox-
ylate group of the uronic acid glycosides 1 and 2 on the
biological activity, therefore their D-glucopyranose 7
and L-idopyranose analogues 10—containing an O-sul-
fate instead of the carboxylate group at the terminal po-
sition—were synthesized using the pathway in Scheme 1.
Glycosidation of 2,5-anhydro-1,6-di-O-benzyl-D-manni-
tol 49,11,12 with acetobromoglucose 3 using mercury
cyanide as promoter afforded after column chromato-
graphy glycoside 5 in modest yield (30%). This mixed
´
ester was submitted to Zemplen deacylation and the
obtained crystalline disaccharide 6 was converted with
DMFÆSO3 into its hepta-O-sulfate, which after treat-
ment with sodium acetate afforded the corresponding
hepta sodium salt 7. The corresponding L-idopyranoside
q Poly-O-sulfated saccharides as IP3 receptor antagonists. Part I.
*
Corresponding author. Tel.: +36 1 399 3300; fax: +36 1 399 3356;
e-mail: janos.kuszmann@idri.hu
0008-6215/$ - see front matter ꢀ 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carres.2005.05.006

1740
J. Kuszmann et al. / Carbohydrate Research 340 (2005) 1739–1749
RO
1
RO
O
NaOOC
RO
NaOOC
O
O
O
O
1'
O
RO
RO
OR
OR
OR
OR
OR
RO
OR
2 R = SO3Na
1 R = SO3Na
Figure 1.
R1O
AcO
AcO
1
R3O
R3O
O
O
OBz
O
BzO
O
Br
1'
O
+
HO
OH
OAc
AcO
OR2
OR1
OR3
R3O
4
3
5 R1 = Bz, R2 = H, R3 = Ac
6 R1 = R2 = R3 = H
7 R1 = R2= R3 = SO3Na
RO
1
BzO
O
RO
RO
HO
O
O
O
1'
O
O
HO
OR
OR
OR
RO
OBz
HO
OAc
HO
8
9 R = H
10 R = SO3Na
11 R = SO3K
Scheme 1.
isomer 10 was synthesized starting from the mixed ester
89 by deacylation and subsequent sulfation of the
obtained amorphous disaccharide 9 (Scheme 1). It is
worth mentioning that while the IP3 antagonist activity
decreased in the case of the D-gluco derivatives by
changing the carboxylate for an O-sulfonate group
(IC50: 1.28 lM (1) ! 4.81 lM (7)), a reversed tendency
could be observed in the L-ido isomers (IC50: 2.35 lM
(2) ! 1.49 lM (10)). As the potassium salts of polysulf-
ated saccharides are usually easier to handle than the
sodium salts,13 the polysulfated idopyranoside was con-
verted into the hepta potassium salt 11 too. As this
change did not influence the biological activity (IC50:
1.39 lM), all further derivatives were isolated as their
potassium salts.
chromatography, after further O-benzoylation. The so
obtained tri-O-benzoates 13 and 17 were converted after
deacylation (14 and 18) into the potassium salts of their
hexa-O-sulfates 15 and 19, respectively (Scheme 2).
These isomers showed very weak biological activity
(IC50: 20.33 lM (15) and 26.53 lM (19)), which led to
the conclusion that at least seven negative charges are
needed for a molecule to act as a potent IP3 antagonist.
For proving this hypothesis, the number of the O-sulfate
groups was increased by attaching a disaccharide to the
aglycon, using acetobromomaltose 20 as glycosylating
agent. The obtained glycoside 21 was converted via
deacylation (22) and subsequent sulfation into the deca
potassium salt 23, which showed a substantially in-
creased biological activity (IC50: 0.22 lM). A trisaccha-
ride derivative 27 with similar activity (IC50: 0.24 lM)
was obtained when the same reaction sequence
(24 ! 25 ! 26 ! 27) was repeated using gentiobiopyr-
anosyl bromide 24 as glycosylating agent. (Scheme 3).
That means that the stereochemistry as well as the place
Next, we used pentopyranoside derivatives, that is,
acetobromo L- (12) and D-arabinopyranose (16), as gly-
cosylating agents. In both cases, multicomponent mix-
tures were obtained from which the needed glycosides
could only be separated in modest yields, by column

J. Kuszmann et al. / Carbohydrate Research 340 (2005) 1739–1749
1741
R1O
1
O
O
O
1'
R2O
O
Br
AcO
4
+
+
OR1
OR1
OR2
R2O
OAc
AcO
13 R1 = Bz, R2 = Ac
14 R1 = R2 = H
15 R1 = R2= SO3K
12
R1O
1
O
O
O
1'
R2O
O
Br
AcO
4
OR1
OR1
OR2
R2O
OAc
AcO
17 R1 = Bz, R2 = Ac
18 R1 = R2 = H
16
19 R1 = R2 = SO3K
Scheme 2.
R1O
1
AcO
R3O
R3O
R3O
AcO
AcO
O
O
O
O
O
1'
1''
4
Br
O
+
O
O
OR2
OR1
R3O
OR3
R3O
OAc
OR3
AcO
OAc
20
AcO
21 R1 = Bz, R2 = H, R3 = Ac
22 R1 = R2 = R3 = H
23 R1 = R2= R3 = SO3K
R3O
R3O
AcO
O
1''
R1O
O
O
1
O
AcO
4
OR3
R3O
O
R3O
O
1'
+
O
O
AcO
AcO
Br
AcO
OR2
OR1
OR3
R3O
OAc
AcO
25 R1 = Bz, R2 = H, R3 = Ac
26 R1 = R2 = R3 = H
24
27 R1 = R2= R3 = SO3K
Scheme 3.
of the glycosidic bond of the disaccharide unit attached
to 2,5-anhydro-D-mannnitol (a-(1!4) in 23 vs b-(1!6)
in 27) has practically no influence on the activity. This
rose the question of whether the number of the charges,
or the place of the attachment of the glycosylating unit
to the 2,5-anhydro-D-mannitol residue is more impor-
tant for the biological activity. Accordingly, the synthe-
sis of a trisaccharide, containing two glucopyranosyl
units attached to the two primary OH groups of the cen-
tral 2,5-anhydro-mannitol moiety, was carried out in the
following way. 2,5-Anhydro-D-mannitol was converted
via its 1,6-di-O-trityl derivative 28 into the correspond-
ing dibenzoate 29, the trityl groups of which were split
off yielding 30 and the latter was glycosylated using aceto-
bromoglucose 3 as donor. The obtained trisaccharide 31
gave after deacylation (32) and subsequent sulfation the

1742
J. Kuszmann et al. / Carbohydrate Research 340 (2005) 1739–1749
TrO
OTr
RO
OR
O
O
3
+
OH
HO
OBz
BzO
28
29 R = Tr
30 R = H
R1O
OR1
OR1
1
6
O
O
O
1'
1'
R1O
O
O
OR2
R2O
R1O
OR1
OR1
R1O
31 R1 = Ac, R2 = Bz
32 R1 = R2 = H
33 R1 = R2 = SO3K
R1O
R1O
AcO
AcO
OR1
OR1
O
1
6
O
O
O
SPh
1'
1'
30
+
O
O
OR3
R3O
OAc
R2O
BnO
OR1
OR1
R1O
34
35 R1 = Ac, R2 = Bn, R3 = Bz
36 R1 = R3 = H, R2 = Bn
37 R1 = R2 = R3 = H
38 R1 = R2 = R3 = SO3K
Scheme 4.
deca potassium salt 33, which was only half as active
(IC50: 0.54 lM) as the isomeric trisaccharides 23 and
27. The activity decreased further (IC50: 1.40 lM), when
instead of D-glucopyranose L-idopyranose was attached
to the primary OH groups of 2,5-anhydro-D-mannitol
(38) (Scheme 4). This led to the conclusion that, besides
the number of the negative charges, the shape of the
molecule is also strongly influencing the biological activ-
ity. For proving this assumption a molecule, with iden-
tical number of charges but differing in the sequence of
the sugar units, was synthesized attaching maltose to
one of the primary OH groups of the anhydro-mannitol
moiety. This derivative was synthesized using thiophen-
ylglycoside 39 as donor and the 3,4-di-O-benzoate 30 as
acceptor. During the glycosidation reaction, a partial O-
acetyl migration from the donor molecule to the accep-
tor took place, therefore the disaccharide 40, which
could be isolated after column chromatography in a
very modest yield only (33%), contained a 6-O-acetyl
group on the 2,5-anhydro moiety. The disaccharide 40
was converted the usual way into the deca potassium
salt 42 of the polysulfated compound (Scheme 5). This
polysulfated trisaccharide possessed a remarkable bio-
logical activity (IC50: 0.11 lM).
For increasing further the number of the O-sulfate
groups, a ꢀbranched-chainꢁ tetrasaccharide 46 was syn-
thesized by glycosylating the unprotected 2,5-anhydro-
D-mannitol 43 with acetobromoglucose 3 followed by
the usual processing (Scheme 6). As expected, this com-
pound showed a very strong biological activity (IC50:
0.14 lM). Finally, two linear polysulfated pentasaccha-
rides were synthesized, using the 3,4-dibenzoate 30 as
acceptor and maltosyl bromide 20 as well as gentiobio-
syl bromide 24 as donors. The formed two isomeric pen-
ta- saccharide derivatives 47 and 50 were converted after
deprotection (48 and 51) and subsequent sulfation into
the corresponding hexadeca potassium salts 49 and 52
(Scheme 7). Both proved to be strong IP3 antagonists
(IC50: 0.11 and 0.18 lM, respectively), but they pos-
sessed some antithrombotic activities too, which is an
unfavourable side effect in the case of antiasthmatic
compounds. From these biological results, the conclu-
sion can be drawn that at least seven negative charges/
molecule are needed for the IP3 antagonist activity,
which becomes stronger when the number of the charges
is increased. However, there seems to be threshold to
this process, as when a certain molecule-size is reached,
the antithrombotic activity becomes dominant.

J. Kuszmann et al. / Carbohydrate Research 340 (2005) 1739–1749
1743
AcO
O
AcO
AcO
O
O
SPh
30
+
OAc
AcO
OAc
AcO
39
R1O
R1O
R1O
1
O
O
O
1'
1''
OR1
O
O
R2O
OR2
OR1
R1O
OR13
R1O
40 R1 = Ac, R2 = Bz
41 R1 = R2 = H
42 R1 = R2 = SO3K
Scheme 5.
R1O
R1O
OR1
OR1
O
6
1
O
O
O
OH
HO
1'
1'
O
O
3
3
+
HO
OR2
OH
R1O
OR1
OR1
R1O
O
43
O
1'
OR1
R1O
R1O
44 R1 = Ac, R2 = H
45 R1 = R2 = H
OR1
46 R1 = R2 = SO3K
Scheme 6.
1. Experimental
selective 1D TOCSY and NOESY spectra were
recorded.
1.1. Generalmethods
1.2. Generalprocedure A for the gyl cosidation reaction
Organic solns were dried over MgSO4 and concentrated
under diminished pressure at or below 40 ꢁC. TLC: E.
Merck precoated Silica Gel 60 F254 plates, with EtOAc
(A), EtOAc–hexane mixtures (B, 1:1; C, 1:2; D, 1:5; E,
2:1; F, 5:1) and EtOAc–MeOH 1:5 (G); detection by
spraying the plates with a 0.02 M soln of I2 and a
0.3 M soln of KI in 10% aq H2SO4 soln followed by
heating at ca. 200 ꢁC. For column chromatography,
Kieselgel 60 was used. The mp are uncorrected. Optical
rotations were determined on 1.0% solns in CHCl3 at
20 ꢁC unless stated otherwise. The NMR spectra were
recorded on a Bruker Avance 500 spectrometer at 500
(1H) and 125 (13C) MHz, respectively, at ambient tem-
perature. The chemical shifts were referenced to
A soln of the acceptor molecule (10 mmol) in dry MeCN
(100 mL) was stirred in the presence of molecular sieves
˚
(4 A) (7 g) for 30 min. Thereafter, the acetobromo donor
(12 mmol) and Hg(CN)2 (13 mmol) were added and stir-
ring was continued at rt for 20 h. The filtered mixture
was diluted with 3-fold CHCl3, washed with 5% Na-
HCO3 soln and a 10% aq soln of KBr, dried and
concentrated.
1.3. Generalprocedure B for the deacyal tion
To a soln of the glycoside (10 mmol) obtained according
to procedure A in dry MeOH (100 mL), 2 M methanolic
MeONa (1 mL) was added. When according to TLC the
deacylation was complete, the sodium ions were
removed with an ion-exchange resin and the residue
d
TMS = 0 ppm. The solvent is indicated at the
1H NMR spectral data. For structure determination,
1H,1H COSY, TOCSY, HMQC, HMBC as well as

1744
J. Kuszmann et al. / Carbohydrate Research 340 (2005) 1739–1749
20
30
+
R1O
R1O
OR1
OR1
R1O
O
OR1
O
O
O
1
6
O
O
O
1''
1''
1'
1'
O
O
OR2
R2O
R1O
OR1
OR1
R1O
R1O
OR1
OR1
R1O
47 R1 = Ac, R2 = Bz
48 R1 = R2 = H
49 R1 = R2 = SO3K
R1O
R1O
OR1
OR1
24
30
+
O
O
1''
1''
O
O
R1O
OR1
O
OR1
6
R1O
1
O
O
1'
1'
R1O
O
OR1
O
OR2
R2O
R1O
OR1
OR1
R1O
50 R1 = Ac, R2 = Bz
51 R1 = R2 = H
52 R1 = R2 = SO3K
Scheme 7.
obtained on concentration was dissolved in water
(50 mL) and freeze-dried.
ion-exchange resin (Na+). The obtained soln was con-
centrated and the residue filtered with MeOH. The solid
material was stirred with MeOH (20 mL) overnight, fil-
tered, washed with MeOH and dried over P2O5.
1.4. Generalprocedure C for the O-suflation
To a stirred and cooled (À20 ꢁC) suspension of
SO3ÆDMF (15 mmol) in DMF (5 mL), a soln of the sac-
charide to be sulfated (containing 10 mmol free OH
groups) in DMF (4 mL) was added at such a rate to
keep the temperature below À15 ꢁC. Thereafter, the
temperature was raised to 0 ꢁC, after 1 h the mixture
was cooled again to À15 ꢁC and EtOH (1 mL) was
added while keeping the temperature below À10 ꢁC.
The obtained mixture was processed according to the
following general procedures.
1.6. Generalprocedure E for the preparation of the
potassium salts
The mixture obtained according to procedure C was
poured into a vigorously stirred and cooled (0 ꢁC) soln
of KOAc (4 g) in MeOH (40 mL). The formed precipi-
tate was filtered off, washed with MeOH (3 · 40 mL)
and dissolved in water (60 mL). The pH of the soln
was adjusted to 8 with M KOH, and M Sr(OAc)2 was
added. The formed precipitate was filtered off and the
filtrate was submitted to a column of CHELX 100
ion-exchange resin (K+). The obtained soln was concen-
trated to a volume of 10 mL and kept overnight at
+4 ꢁC. The precipitated material was filtered, washed
with cold (0 ꢁC) water and dried over P2O5.
1.5. Generalprocedure D for the preparation of the
sodium salts
The mixture obtained according to procedure C was
poured into a vigorously stirred and cooled (0 ꢁC) soln
of NaOAc (4 g) in MeOH (40 mL). The formed precip-
itate was filtered off, washed with MeOH (3 · 40 mL)
and dissolved in water (60 mL). The pH of the soln
was adjusted to 8 with M NaOH, and M Sr(OAc)2
was added. The formed precipitate was filtered off and
the filtrate was submitted to a column of CHELX 100
1.7. 2,5-Anhydro-3-O-(2,3,4,6-tetra-O-acetyl-b-D-gluco-
pyranosyl)-1,6-di-O-benzoyl-D-mannitol(5)
Prepared according to the general procedure A from 4
(9.0 g, 24 mmol) and 3 (10.5 g, 25.5 mmol) to yield after
column chromatography (B) 5 (5.4 g, 30%) as syrup:

J. Kuszmann et al. / Carbohydrate Research 340 (2005) 1739–1749
1745
1
[a]D +46 (c 1, CHCl3); Rf 0.4. Anal. Calcd for
C34H38O16: C, 58.12; H, 5.45. Found: C, 58.01; H, 5.56.
À4 (c 1, H2O); H NMR and 13C NMR identical with
10; Anal. Calcd for C12H15O31S7K7: C, 12.50; H, 1.31;
S, 16.46; K 23.73. Found: C, 12.38; H, 1.82; S, 18.50;
K, 22.90.
1.8. 2,5-Anhydro-3-O-(-b-D-glucopyranosyl)-D-mannitol
(6)
1.13. 2,5-Anhydro-3-O-(2,3,4-tri-O-acetyl-a-L-arabino-
pyranosyl)-1,4,6-tri-O-benzoyl-D-mannitol(13)
Prepared according to the general procedure B from 5
(5.2 g, 7.4 mmol) to yield 6 (1.7 g, 71%); mp 181–
183 ꢁC (MeOH), [a]D +20 (c 1, water). Anal. Calcd for
C12H22O10: C, 44.17; H, 6.80. Found: C, 43.89; H, 6.92.
Prepared according to the general procedure A from 12
(12 g, 35.4 mmol) and 4 (13.0 g, 35 mmol) as starting
materials. The residue of the concentrated CHCl3 soln
was dissolved in pyridine (100 mL) and benzoyl chloride
(15 mL) was added. The mixture was stirred at rt for 2 h,
then poured into ice-water. The precipitated oil was dis-
solved in CH2Cl2 to give after usual processing a syrup,
which was purified by column chromatography (C).
Concentration of the fractions having Rf 0.4 gave 13
(11 g, 39%) as syrup; [a]D +12 (c 1, CHCl3). Anal. Calcd
for C38H38O15: C, 62.12; H, 5.21. Found: C, 62.34; H,
5.32.
1.9. 2,5-Anhydro-1,4,6-tri-O-sulfato-3-O-(2,3,4,6-tetra-
O-sulfato-b-D-glucopyranosyl)-D-mannitolhepta sodium
salt (7)
Prepared according to the general procedure C and D
from 6 (0.65 g, 2 mmol) to give 7 (1.93 g, 93%); [a]D
+15 (c 1, water); 1H NMR: d 4.94–4.89 (m, 2H, H-
4,10), 4.69 (m, 1H, H-30), 4.55–4.33 (m, 6H, H-
2,3,5,20,40,6a0), 4.27–4.12 (m, 5H, H-1a,1b,6a,6b,6b0),
13
4.07 (m, 1H, H-50); C NMR: d 102.9 (C-10), 86.7,
84.5, 84.4, 83.6 (C-2,3,4,5), 79.5, 79.2 (C-20,30), 76.4,
75.4 (C-40,50), 70.1, 69.7, 69.7 (C-1,6,60). Anal. Calcd
for C12H15O31S7Na7: C, 13.85; H, 1.45; Na, 15.47; S,
21.57. Found: C, 13.99; H, 1.65; Na, 15.39; S, 21.40.
1.14. 2,5-Anhydro-3-O-(-a-L-arabinopyranosyl)-D-
mannitol(14)
Prepared according to the general procedure B from 13
(10.9 g, 14.8 mmol) to give 14 as hygroscopic amor-
phous powder (4.1 g, 94%), [a]D +67 (c 1, water). Anal.
Calcd for C11H20O9: C, 44.59; H, 6.80. Found: C, 44.62;
H, 6.95.
1.10. 2,5-Anhydro-3-O-(a-L-idopyranosyl)-D-mannitol(9)
Prepared according to the general procedure B from 89
(1.65 g, 3.97 mmol) to yield 9 (1.06 g, 82%); mp 162–
164 ꢁC (c 1, MeOH), [a]D À40 (c 1, water). Anal. Calcd
for C12H22O10: C, 44.17; H, 6.80. Found: C, 44.00; H,
6.85.
1.15. 2,5-Anhydro-1,4,6-tri-O-sulfato-3-O-(2,3,4-tri-O-
sulfato-a-L-arabinopyranosyl)-D-mannitolhexa
potassium salt (15)
1.11. 2,5-Anhydro-1,4,6-tri-O-sulfato-3-O-(2,3,4,6-tetra-
O-sulfato-a-L-idopyranosyl)-D-mannitolhepta sodium salt
(10)
Prepared according to the general procedure C and D
from 14 (0.59 g, 2 mmol) to give 15 (1.6 g, 80%); [a]D
+33 (c 1, water); 1H NMR: d 4.88 (t, 1H, J3,4
=
Prepared according to the general procedure C and D
from 9 (0.65 g, 2 mmol) to give 10 (1.8 g, 87%); [a]D
J4,5 = 3 Hz, H-4), 4.87–4.81 (m, 2H, H-10,40), 4.68 (m,
1H, H-20), 4.58–4.43 (m, 3H, H-3,5,30), 4.41 (m, 1H,
H-2), 4.30–4.15 (m, 5H, H-1a,1b,6a,6b,5a0), 3.76 (dd,
1
À3 (c 1, water); H NMR: d 5.23 (s, 1H, H-10), 5.03
(s, 1H, H-30), 4.86 (t, 1H, J2,3 = J3,4 = 2.5 Hz, H-4),
1H, J5a ,5b = 12.7, J5b ,4 = 2.5 Hz, H-5b0); 13C NMR: d
102.6 (C-10), 86.2 (C-3), 84.7 (C-4), 84.0 (C-5), 82.9
(C-2), 77.0 (C-30), 76.6 (C-20), 74.4 (C-40), 70.0 (C-1),
69.4 (C-6), 63.2 (C-50). Anal. Calcd for C11H14O27S6K6:
C, 13.14; H, 1.40; S, 19.14; K 23.34. Found: C, 13.05; H,
1.78; S, 18.55; K, 23.30.
0
0
0
0
4.60–4.46 (m, 6H, H-2,3,5,20,40,50), 4.31 (dd, 1H,
J6a ,6b = 11.2, J6a ,5 = 3.4 Hz, H-6a0), 4.25–4.14 (m,
0
0
0
0
13
5H, H-1a,1b,6a,6b,6b0); C NMR: d 100.1 (C-10),
84.5, 84.4, 84.4, 83.2 (C-2,3,4,5), 73.4, 72.9, 72.8, 66.7
(C-20,30,40,50), 70.0, 69.8, 69.3 (C-1,6,60). Anal. Calcd
for C12H15O31S7Na7: C, 13.85; H, 1.45; Na, 15.47; S,
21.57. Found: C, 14.03; H, 1.69; Na, 15.32; S, 21.32.
1.16. 2,5-Anhydro-3-O-(2,3,4-tri-O-acetyl-a-D-arabino-
pyranosyl)-1,4,6-tri-O-benzoyl-D-mannitol(17)
1.12. 2,5-Anhydro-1,4,6-tri-O-sulfato-3-O-(2,3,4,6-tetra-
O-sulfato-a-L-idopyranosyl)-D-mannitolhepta potassium
salt (11)
Acetobromo D-arabinose (16) was coupled with 4 as de-
scribed for the L-isomer 13 to give 17 (10.6 g, 38%); [a]D
0 (c 1, CHCl3). Anal. Calcd for C38H38O15: C, 62.12; H,
5.21. Found: C, 62.28; H, 5.41.
Prepared according to the general procedure C and E
from 9 (0.65 g, 2 mmol) to give 11 (2.05 g, 89%); [a]D

1746
J. Kuszmann et al. / Carbohydrate Research 340 (2005) 1739–1749
1.17. 2,5-Anhydro-3-O-(-a-D-arabinopyranosyl)-D-
mannitol(18)
1,6,60,600). Anal. Calcd for C18H22O35S10K10: C, 12.95;
H, 1.33; S, 19.20; K, 23.41. Found: C, 12.48; H, 1.65;
S, 18.95; K, 23.08.
Deacylation of 17 (8.4 g, 11.5 mmol) was carried out as
described for the L-isomer 14 to give 18 (3.1 g, 91%) as
an amorphous powder; [a]D +7 (c 1, water). Anal. Calcd
for C11H20O9: C, 44.59; H, 6.80. Found: C, 44.42; H,
6.95.
1.22. 2,5-Anhydro-1,6-di-O-benzoyl-3-O-(heptaacetyl-b-
gentiobiosyl)-D-mannitol(25)
Prepared according to the general procedure A from 4
(2.6 g, 7 mmol) and 24 (4.7 g, 6.5 mmol) to give 25
(2.2 g, 32%); mp 166–168 ꢁC (EtOH), [a]D +9 (c 1,
CHCl3). Anal. Calcd for C46H54O240: C, 55.76; H,
5.49. Found: C, 55.79; H, 5.51.
1.18. 2,5-Anhydro-1,4,6-tri-O-sulfato-3-O-(2,3,4-tri-
O-sulfato-a-D-arabinopyranosyl)-D-mannitolhexa
potassium salt (19)
Sulfation of 18 (0.59 g, 0.3 mmol) was carried out as de-
scribed for the L-isomer 15 to give 19 (1.8 g, 90%); [a]D
1.23. 2,5-Anhydro-3-O-(b-gentiobiosyl)-D-mannitol(26)
1
+21 (c 1, water); H NMR: d 5.02 (m, 1H, H-10), 4.90–
Prepared according to the general procedure B from 25
(2.0 g, 1.87 mmol) to give 26 (0.95 g, ꢀ100%) as
amorphous powder, [a]D +7 (c 1, water). Anal. Calcd
for C18H32O15: C, 44.26; H, 6.60. Found: C, 44.14; H,
6.85.
4.78 (m, 2H, H-40,40), 4.75 (m, 1H, H-20), 4.60–4.55 (m,
2H, H-5,30), 4.52 (m, 1H, H-3), 4.42 (m, 1H H-20), 4.32–
4.15 (m, 4H, H-1a,1b,6a,6b), 4.12 (m, 1H, H-5a0), 3.78
13
(m, 1H, H-5b0); C NMR: d 100.8 (C-10), 84.8 (C-4),
84.2 (C-5), 84.1 (C-3), 82.8 (C-2), 76.6 (C-30), 76.3 (C-
20), 74.2 (C-40), 69.7 (C-6), 69.3 (C-1), 62.5 (C-50). Anal.
Calcd for C11H14O27S6K6: C, 13.14; H, 1.40; S, 19.14; K,
23.34. Found: C, 13.00; H, 1.69; S, 18.63; K, 23.21.
1.24. 2,5-Anhydro-1,4,6-tri-O-sulfato-3-O-[6-O-(2,3,4,6-
tetra-O-sulfato-a-D-glucopyranosyl)-2,3,4-tri-O-sulfato-
b-D-glucopyranosyl]-D-mannitoldeca potassium satl (27)
1.19. 2,5-Anhydro-1,6-di-O-benzoyl-3-O-(hepta-O-acetyl-
b-D-maltosyl)-D-mannitol(21)
Prepared according to the general procedure C and D
from 26 (0.54 g, 1.1 mmol) to give 27 (1.75 g, 95%),
[a]D +7 (c 1, water); 1H NMR: d 4.97 (d, 1H,
Prepared according to the general procedure A from 4
(6.5 g, 17.5 mmol) and 20 (12.5 g, 18.4 mmol) to yield
after column chromatography (B) 21 as a syrup (5.6 g,
30%), Rf 0.4; [a]D +65 (c 1, CHCl3). Anal. Calcd for
C46H54O240: C, 55.76; H, 5.49. Found: C, 55.89; H, 5.62.
J1 ,2 = 7.4 Hz, H-100), 4.91 (t, 1H, J3,4 = J4,5 = 4 Hz,
00 00
H-4), 4.83 (d, 1H J1 ,2 = 6.3, H-10), 4.76 (t, 1H,
0
0
= 8.7 Hz, H-40) 4.69 (t, 1H, J2 ,3
=
0
J3 ,4
0
0
0
00 00
=
J4 ,5
J3 ,4 = 8.3 Hz, H-300), 4.61 (t, 1H, J2,3 = 4 Hz, H-3),
4.59 (t, 1H, J2,3 = 4 Hz, H-30) 4.53 (m, 2H, H-5,600)
4.47–4.00 (m, 11H), 3.90 (m, 1H, H-50); 13C NMR: d
104.1 (C-100), 103.1 (C-10), 86.4 (C-3), 84.6 (C-4), 83.2
(C-5), 82.6 (C-2), 80.7 (C-30), 79.9 (C-300), 79.6 (C-200),
79.3 (C-20), 76.7 (C-50), 76.4 (C-400), 76.0 (C-40) 75.5
(C-500), 71.1 (C-60), 70.4 (C-6), 70.0 (C-1), 70.1 (C-600).
Anal. Calcd for C18H22O45S10K10: C, 12.90; H, 1.32;
S, 19.18; K, 23.38. Found: C, 12.59; H, 1.89; S, 18.33;
K, 21.08.
00 00
1.20. 2,5-Anhydro-3-O-(b-D-maltosyl)-D-mannitol(22)
Prepared according to the general procedure B from 21
(1.6 g, 1.6 mmol) to give 22 (0.75 g, 95%) as an
amorphous powder, [a]D +96 (c 1, water). Anal. Calcd
for C18H32O15: C, 44.26; H, 6.60. Found: C, 44.08; H,
6.82.
1.21. 2,5-Anhydro-1,4,6-tri-O-sulfato-3-O-[4-O-(2,3,4,6-
tetra-O-sulfato-a-D-glucopyranosyl)-2,3,6-tri-O-sulfato-
b-D-glucopyranosyl]-D-mannitoldeca potassium satl (23)
1.25. 2,5-Anhydro-1,6-di-O-trityl-D-mannitol(28)
To a stirred soln of crude 2,5-anhydro-D-mannitol11,14
(4.6 g, 28 mmol) in pyridine (50 mL), TrCl (23 g,
70 mmol) was added and the mixture was stirred for
2 days at 20 ꢁC. Thereafter, it was poured into water, ex-
tracted with CH2Cl2, washed with M H2SO4, water, 5%
NaHCO3 and water, dried and concentrated. The semi-
crystalline residue (26 g) was purified by column chroma-
tography using first 1:5 EtOAc–hexane for removing the
by-products (Rf 0.9; 0.7 and 0.5) and then EtOAc for
eluting the product. Concentration of this latter fraction
gave 28 (13.2 g, 74%), Rf 0.5 (B); [a]D +14 (c 1, pyridine)
as solid foam, which was used without further
Prepared according to the general procedure C and D
from 22 (0.6 g, 1.2 mmol) to give 23 (1.95 g, 95%),
[a]D +33 (c 1, water); 1H NMR: d 5.59 (d, 1H,
J1 ,2 = 3.1 Hz, H-100), 5.04 (d, 1H, J1 ,2 = 5.3 Hz, H-
00 00
0
0
10), 4.95 (s, 1H, H-4), 4.86 (t, 1H, J2 ,3 = J3 ,4 = 8 Hz,
00 00
00 00
H-300) 4.75 (t, J2 ,3 = 5 Hz, H-20), 4.58 (s, 1H, H-3),
0
0
13
4.56–4.03 (m, 16H); C NMR: d 103.7 (C-10), 97.3
(C-100), 86.8 (C-3), 84.8 (C-4), 84.5, 83.5 (C-2,5), 79.9
(C-20), 79.4 (C-200), 77.7, 76.5, 76.2, 76.1, 74.9 (C-40,
C-50, C-300, C-400, C-500), 70.2, 70.0, 69.8, 68.9 (C-

J. Kuszmann et al. / Carbohydrate Research 340 (2005) 1739–1749
1747
purification in the next step. Lit.15 mp 78–85 ꢁC, [a]D
+18 (c 1, pyridine), yield 15.5%.
1.32; S, 19.18; K, 23.38. Found: C, 11.66; H, 1.67; S,
18.68; K, 22.92.
1.26. 2,5-Anhydro-3,4-di-O-benzoyl-D-mannitol(30)
1.30. 2,5-Anhydro-1,6-bis-O-(3-O-benzyl-a-L-
ido-pyranosyl)-D-mannitol(36)
To a stirred soln of 28 (13.0 g, 20 mmol) in pyridine
(60 mL), benzoyl chloride (6.0 mL, 50 mmol) was
added gradually with cooling below 10 ꢁC. The mixture
was kept at rt for 3 h, then poured into water and ex-
tracted with CH2Cl2. The organic soln was processed
the usual way to give on concentration crude 29
(17.0 g, ꢀ100%). This was dissolved in AcOH
(100 mL) at 80 ꢁC and water (25 mL) was added grad-
ually. The soln was kept at this temperature for 1 h,
and TrOH, which precipitated on cooling, was filtered
off. The filtrate was diluted with CH2Cl2, washed with
water, 5% NaHCO3 soln and water, dried and
concentrated. The semisolid residue was filtered with
toluene, to give after recrystallization from 3-fold
toluene 30 (2.6 g, 35%), mp 120–122 ꢁC. Anal. Calcd
for C20H20O7: C, 64.51; H, 5.41. Found: C, 54.50; H,
5.44.
To a stirred soln of 2,5-anhydro-3,4-di-O-benzoyl-D-
mannitol 30 (4.46 g, 12 mmol) in CH2Cl2 (150 mL),
˚
molecular sieve 4 A (18 g) and the thiophenyl glycoside
349 (13 g, 27 mmol) were added. After 30 min the mix-
ture was cooled to À40 ꢁC, NIS (9 g, 40 mmol) and
TfOH (0.5 mL) were added and stirring was continued
at À40 ꢁC for 15 min. Thereafter, Et3N (9 mL) was
added and the temperature was raised to 20 ꢁC. The fil-
tered mixture was washed with aq Na2S2O3 and subse-
quently with NaHCO3 soln and water. The dried
organic soln was concentrated and the residue purified
by column chromatography (B). Concentration of the
fractions having Rf 0.35 gave crude 2,5-anhydro-3,4-
di-O-benzoyl-1,6-bis-O-(2,4,6-tri-O-acetyl-3-O-benzyl-
a-L-idopyranosyl)-D-mannitol 35 (8 g, 59%) as syrup,
[a]D À75 (c 1, CHCl3) the purity of which was ꢀ60%
according to NMR. This was dissolved in MeOH
(80 mL), and 2 M NaOMe in MeOH (0.3 mL) was
added. After 2 h, when according to TLC the deacy-
lation was complete the sodium ions were removed with
an ion-exchange resin and the filtered soln was concen-
trated. The residue was partitioned between water and
CH2Cl2 for removing the methyl benzoate. The aq soln
was freeze-dried to give after column chromatography
(ethyl acetate–ethanol 5:1) 36 (1.7 g, 36%), [a]D À41
(c 1, water).
1.27. 2,5-Anhydro-3,4-di-O-benzoyl-1,6-bis-O-(2,3,4,6-
tetra-O-acetyl-b-D-glucopyranosyl)-D-mannitol(31)
Prepared according to the general procedure A from 30
(2.6 g, 7 mmol) and 3 (10.5 g, 25.5 mmol) to yield after
column chromatography (E) 31 (2.0 g, 28%); mp 132–
134 ꢁC (EtOH), [a]D À28 (c 1, CHCl3). Anal. Calcd
for C46H54O24: C, 55.76; H, 5.49. Found: C, 55.81; H,
5.52.
1.28. 2,5-Anhydro-1,6-bis-O-(b-D-glucopyranosyl)-
D-mannitol(32)
1.31. 2,5-Anhydro-1,6-bis-O-a-L-idopyranosyl-D-
mannitol(37)
Prepared according to the general procedure B from 31
(1.6 g, 1.55 mmol) to give 32 (0.75 g, ꢀ100%) as
amorphous powder, [a]D À3 (c 1, water). Anal. Calcd
for C18H32O15: C, 44.26; H, 6.60. Found: C, 44.15; H,
6.72.
A soln of 36 (1.7 g) in MeOH (50 mL) and water (5 mL)
was hydrogenated in the presence of 10% Pd/C (1 g) at
normal pressure for 6 h when according to TLC both
benzyl groups were removed. The filtered soln was con-
centrated, the residue dissolved in water (10 mL) and
freeze-dried to yield 37 (1.2 g, 96%) as amorphous pow-
der; [a]D 0 (c 1, water). Anal. Calcd for C18H32O15: C,
44.26; H, 6.60. Found: C, 44.11; H, 6.80.
1.29. 2,5-Anhydro-3,4-di-O-sulfato-1,6-bis-O-(2,3,4,6-
tetra-O-sulfato-a-D-glucopyranosyl)-D-mannitoldeca
potassium salt (33)
1.32. 2,5-Anhydro-3,4-di-O-sulfato-1,6-bis-O-(2,3,4,6-
tetra-O-sulfato-a-L-idopyranosyl)-D-mannitoldeca
potassium salt (38)
Prepared according to the general procedure C and E
from 32 (0.54 g, 1.1 mmol) to give 33 (1.5 g, 83%), [a]D
+33 (c 1, water); 1H NMR: d 5.03 (m, 2H, H-3,4),
4.93 (d, 2H, J1 ,2 = 5.5 Hz, H-10), 4.73 (t, J2 ,3
=
0
Prepared according to the general procedure C and E
from 37 (0.97 g, 2 mmol) to give 38 (2.5 g, 75%), [a]D
0
0
0
J3 ,4 = 6.4 Hz, 2H, H-30), 4.52 (m, 2H, H-40) 4.50–4.38
(m, 6H, H-2,5,20,6a0), 4.26–4.15 (m, 4H, H-1a,6a,6b0),
4.10 (m, 2H, H-50), 3.96–3.87 (m, 2H, H-1b,6b); 13C
NMR: d 103.7 (C-10), 83.2 (C-3,4), 83.0 (C-2,5), 78.9
(C-20,30), 76.2 (C-50), 75.6 (C-40), 71.6 (C-1,6), 70.3 (C-
60). Anal. Calcd for C18H22O45S10K10: C, 12.90; H,
0
0
1
À13 (c 1, water); H NMR: d 5.18 (s, 2H, H-10), 5.07
(s, 2H, H-30), 4.99 (s, 2H, H-3,4), 4.63 (s, 2H, H-40)
4.59 (s, 2H, H-20), 4.54 (m, 4H, H-2,5,50), 4.31–4.21
(m, 4H, H-60), 4.05 (d, 2H, J1a,1b,6a,6b = 11.3 Hz, H-
1a,6a), 3.82 (dd, 2H, J1b,2,5,6b = 6.5 Hz, H-1b,6b); 13C

1748
J. Kuszmann et al. / Carbohydrate Research 340 (2005) 1739–1749
NMR: d 101.5 (C-10), 84.0 (C-3,4), 83.8 (C-2,5), 73.4 (C-
20,30), 73.0 (C-40), 70.2 (C-60), 70.1 (C1,6), 66.7 (C-50).
Anal. Calcd for C18H22O45S10K10: C, 12.95; H, 1.33;
S, 19.20; K, 23.41. Found: C, 12.48; H, 1.62; S, 18.89;
K, 22.71.
1.37. 2,5-Anhydro-1,3,6-tri-O-(2,3,4,6-tetra-O-acetyl-b-
D-glucopyranosyl)-D-mannitol(44)
Prepared according to the general procedure A from 43
(1.64 g, 10 mmol) and 3 (14.8 g, 36 mmol) to give after
column chromatography (F) 44 (3.9 g, 34%), Rf 0.4;
[a]D +4 (c 1, CHCl3). Anal. Calcd for C48H66O32: C,
49.91; H, 5.76. Found: C, 49.65; H, 5.83.
1.33. Phenyl1-thio- b-maltopyranosyl-peracetate (39)
Acetobromomaltose, obtained from octaacetyl-maltose
(11.5 g, 17 mmol) according to the lit.,16 was dissolved
in 1,2-dichloroethane (60 mL) and thiophenol (2 mL)
was added followed by Et3N (5 mL). The mixture was
stirred at rt for 2 h, washed with water, concentrated
and the residue submitted to column chromatography
(B). Concentration of the fractions having Rf 0.5 affor-
ded 39 (7 g, 56%) as syrup; [a]D +57 (c 1, CHCl3).
Lit.17 [a]D +47 (c 1, CHCl3).
1.38. 2,5-Anhydro-1,3,6-tri-O-b-D-glucopyranosyl-D-
mannitol(45)
Prepared according to the general procedure B from 44
(3.70 g, 3.2 mmol) to give 45 (1.80 g, 97%), [a]D +8 (c 1,
water). Anal. Calcd for C24H42O20: C, 44.31; H, 6.51.
Found: C, 44.44; H, 6.65.
1.39. 2,5-Anhydro-4-O-sulfato-1,3,6-tri-O-(2,3,4,6-
tetra-O-sulfato-a-D-glucopyranosyl)-D-mannitoltrideca
potassium salt (46)
1.34. 6-O-Acetyl-2,5-anhydro-3,4-di-O-benzoyl-1-O-
[20,30,40,60,2,3,6-hepta-O-acetyl-b-maltopyranosyl]-D-
mannitol(40)
Prepared according to the general procedure C and E
from 45 to yield 46 (1.5 g, 69%), [a]D +4 (c 1, water);
1H NMR: d 5.05–4.88 (m, 4H, H-4,10,100,1000), 4.84–3.87
Prepared as described for 36, starting from 30 (1.6 g,
4.3 mmol) and 39 (7 g, 9.6 mmol), to give after separation
by column chromatography (C) 40 (1.5 g, 33%) as a
syrup, Rf 0.7; [a]D +10 (c 1, CHCl3). Anal. Calcd for
C46H54O240: C, 55.76; H, 5.49. Found: C, 55.859; H, 5.66.
13
(m, 25H); C NMR: d 103.8 (C-10), 85.9, 84.2, 84.2,
82.0 (C-2,3,4,5) 79.4, 79.4, 79.3, 79.1, 78.9, 78.9 (D-glu-
co-2,3), 76.3, 76.2, 76.0, 75.9, 75.8, 75.5 (D-gluco-C-
4,5), 72.5, 70.7, 70.2, 70.2, 69.8 (C-1,6,60,600,6000). Anal.
Calcd for C24H29O59S13K13: C, 13.18; H, 1.34; S,
19.06; K, 23.25. Found: C, 12.38; H, 1.77; S, 18.68; K,
21.85.
1.35. 2,5-Anhydro-1-O-b-maltopyranosyl-D-mannitol(41)
Prepared according to the general procedure B from 40
(1.3 g, 1.2 mmol). The residue obtained on concentra-
tion of the methanolic soln was partitioned between
water and CH2Cl2 for removing the methyl benzoate.
The aq soln was freeze-dried to give 41 (0.6 g, 60%) as
amorphous powder, [a]D +86 (c 1, water). Anal. Calcd
for C18H32O15: C, 44.26; H, 6.60. Found: C, 44.18; H,
6.79.
1.40. 2,5-Anhydro-3,4-di-O-benzoyl-1,6-bis-O-(hepta-O-
acetyl-b-maltosyl)-D-mannitol(47)
Prepared according to the general procedure A from 20
(14 g, 20 mmol) and 30 (2.75 g, 7.4 mmol) to yield after
column chromatography (E) 47 as a syrup (6.3 g, 53%),
Rf 0.35; [a]D +38 (c 1, CHCl3). Anal. Calcd for
C72H88O41: C, 53.73; H, 5.51. Found: C, 53.99; H, 5.82.
1.36. 2,5-Anhydro-3,4,6-tri-O-sulfato-1-O-[4-O-(2,3,4,6-
tetra-O-sulfato-a-D-glucopyranosyl)-2,3,6-tri-O-sulfato-
b-D-glucopyranosyl]-D-mannitoldeca potassium satl (42)
1.41. 2,5-Anhydro-1,6-bis-O-b-maltosyl-D-mannitol(48)
Prepared according to the general procedure C and E
from 41 (0.44 g, 0.9 mmol) to give 42 (1.35 g, 90%),
Prepared according to the general procedure B from 47
(2.1 g) to give after column chromatography (G) 48
(0.7 g, 66%) as a solid foam; Rf 0.4; [a]D +92 (c 1, water).
Anal. Calcd for C30H52O25: C, 44.34; H, 6.45. Found: C,
44.55; H, 6.56.
1
[a]D +31 (c 1, water); H NMR: d 5.56 (s, 2H, H-100),
5.02 (s, 1H, H-10), 4.99 (s, 1H, H-3), 4.96 (s, 1H, H-4),
4.80 (m, 2H, H-30,300) 4.55 (s, 1H, H-20), 4.50 (m, 2H,
H-5,200), 4.48–4.05 (m, 13H), 3.93–3.85 (m, 1H, H-1a);
13C NMR: d 104.0 (C-10), 97.4 (C-100), 84.1 (C-2), 84.0
(C-3), 83.8 (C-4), 83.0 (C-5), 79.1 (C-30), 78.7 (C-20),
77.8 (C-300), 76.6 (C-200), 76.2 (C-400), 75.9 (C-50), 74.9
(C-40), 72.7 (C-500), 71.3 (C-1), 70.4 (C-60), 70.0 (C-6),
69.0 (C-600). Anal. Calcd for C18H22O45S10K10: C,
12.95; H, 1.33; S, 19.20; K, 23.41. Found: C, 12.63; H,
1.68; S, 18.30; K, 22.96.
1.42. 2,5-Anhydro-3,4-di-O-sulfato-1,6-bis-O-[4-O-
(2,3,4,6-tetra-O-sulfato-a-D-glucopyranosyl)-2,3,6-tri-
O-sulfato-b-D-glucopyranosyl]-D-mannitolhexadeca
potassium salt (49)
Prepared according to the general procedure C and E
from 48 (0.63 g, 0.78 mmol) to give 49 (1.5 g, 71%), [a]D

J. Kuszmann et al. / Carbohydrate Research 340 (2005) 1739–1749
1749
+35 (c 1, water); 1H NMR: d 5.56 (s, 2H, H-100), 5.01 (m,
20,30,200,300), 76.2, 76.2, 76.0, 75.6 (C-40,50,400,500), 72.2
(C-1,6), 71.6, 70.4 (C-60,600). Anal. Calcd for
C30H36O73S16K16: C, 13.33; H, 1.33; S, 18.96; K,
23.10. Found: C, 12.62; H, 1.82; S, 17.48; K, 21.41.
4H, H-10,3,4), 4.84 (t, 2H, J2 ,3 = J3 ,4 = 8.3 Hz, H-300),
4.80 (m, 2H, H-30), 4.58–3.86 (m, 26H); 13C NMR: d
104.1 (C-10), 97.5 (C-100), 83.2 (C-3,4), 83.0 (C-2,5), 79.4
(C-30), 78.9 (C-20), 77.7 (C-300), 76.5 (C-200), 76.2, 76.1
(C-50,400), 75.1 (C-40), 72.7 (C-500), 71.4 (C-1,6), 70.3 (C-
60), 69.0 (C-600). Anal. Calcd for C30H36O73S16K16: C,
13.33; H, 1.33; S, 18.96; K, 23.10. Found: C, 12.85; H,
1.72; S, 18.56; K, 22.88.
00 00
00 00
Acknowledgements
´
The authors are very much indebted to Dr. Istvan
Kurucz and Dr. Cecilia Salamon for the biological
results.
1.43. 2,5-Anhydro-3,4-di-O-benzoyl-1,6-bis-O-[6-O-
(2,3,4,6-tetra-O-acetyl-b-D-glucopyranosyl)-2,3,4-tri-O-
acetyl-b-D-glucopyranosyl]-D-mannitol(50)
References
Prepared according to the general procedure A from 24
(4.16 g, 6 mmol) and 30 (1.0 g, 2.7 mmol) to give after
column chromatography (E) 50 (2.1 g, 48%) as syrup,
Rf 0.4; [a]D À17 (c 1, CHCl3). Anal. Calcd for
C72H88O41: C, 53.73; H, 5.51. Found: C, 54.06; H, 5.80.
1. Rabenstein, D. L. Nat. Prod. Rep. 2002, 19, 312–331.
2. Lever, R.; Page, C. Pulm. Pharmacol. Ther. 2001, 14, 249–
254.
3. Diamant, Z.; Page, C. P. Pulm. Pharmacol. Ther. 2000, 13,
1–4.
4. Martineau, P.; Vaugham, L. M. Ann. Pharmacother. 1995,
29, 71–73.
5. Jerzynska, J.; Stelmach, I.; Kuna, P. Pol. Merkuriusz Lek.
2000, 8, 341–346.
1.44. 2,5-Anhydro-1,6-bis-O-[6-O-(b-D-glucopyranosyl)-
b-D-glucopyranosyl]-D-mannitol(51)
6. Ahmed, T.; Broder, S.; Whisnant, J. K. US Pat. No:
5,690,910, 1997; Chem. Abstr. 1977, 126, 233706v.
7. Ahmed, T. US Pat. No: 5,980,865, 1999; Chem. Abstr.
1999, 131, 317774g.
8. Ahmed, T.; Smith, G. A. WO Pat 02/083700, 2002; Chem.
Abstr. 2002, 137, 311148h.
9. Kuszmann, J.; Medgyes, G.; Boros, S. Carbohydr. Res.
2004, 339, 1569–1579.
10. Worley, P. F.; Baraban, J. M.; Supattapone, S.; Wilson, V.
S.; Snyder, S. H. J. Biol. Chem. 1987, 262, 12132–12136.
11. Otero, D. A.; Simpson, R. Carbohydr. Res. 1984, 128, 79–
86.
Prepared according to the general procedure B from 50
(2.0 g, 1.24 mmol). The residue obtained on concentra-
tion of the methanolic soln was partitioned between
water and CH2Cl2 for removing the methyl benzoate.
The aq soln was freeze-dried to give after column chro-
matography (G) 51 (0.80 g, 80%) as solid foam, [a]D À8
(c 1, water). Anal. Calcd for C30H52O25: C, 44.34; H,
6.45. Found: C, 44.62; H, 6.59.
1.45. 2,5-Anhydro-1,6-bis-O-[6-O-(2,3,4,6-tetra-O-
sulfato-a-D-glucopyranosyl)-2,3,4-tri-O-sulfato-b-D-
glucopyranosyl]-D-mannitolhexadeca- O-sulfate
hexadeca potassium salt (52)
12. Lei, Z.; Min, J. M.; Zhang, L. H. Tetrahedron: Asymmetry
2000, 11, 2899–2906.
13. Ochi, K.; Watanabe, Y.; Okui, K.; Shindo, M. Chem.
Pharm. Bull. 1980, 28, 638–641.
14. Horton, D.; Philips, K. D. Methods Carbohydr. Chem. VII
1976, 68–70.
15. Aspinall, G. O.; Przybylski, E.; Ritchie, R. G.; Wong, Ch.
O. Carbohydr. Res. 1978, 66, 225–243.
16. Tropper, F. D.; Andersson, F. O.; Grand-Maitre, C.; Roy,
R. Synthesis 1991, 9, 734–736.
17. Motawia, M. S.; Larsen, K.; Olsen, C. E.; Moeller, B. L.
Synthesis 2000, 11, 1547–1665.
Prepared according to the general procedure C and E
from 51 (0.7 g, 0.86 mmol) to give 52 (1.9 g, 82%), [a]D
1
0 (c 1, water); H NMR: d 5.00 (m, 2H, H-3,4), 4.90–
4.81 (m, 4H, H-10,100), 4.70–4.13 (m, 22H), 4.03–3.89
13
(m, 8H); C NMR: d 104.0, 103.9 (C-1,10), 83.0
(C-3,4), 82.3 (C-2,5), 80.3, 79.9, 79.1, 79.1 (C-