Potent E-selectin antagonists

Article abstract of DOI:10.1002/1522-2675(20001108)83:11<2893::AID-HLCA2893>3.0.CO;2-G

In the search for drugs that could control excessive leukocyte extravasation, we now report on modifications of the already known potent E-selectin antagonist 3 containing a cyclohexyllactic acid residue and a glucal-derived building block. Thus, we describe the synthesis and biological evaluation of a series of derivatives 6 with modified glucal-derived moieties (CH₂NR¹R² instead of CH₂OH in 3) to explore a hypothetical potential complementary interaction with E-selectin. However, similar activity profiles of most derivatives 6 and compound 3 do not support such an interaction, but rather indicate topological-structure changes of 6 (and 3) in the orientation of the neighboring fucose and galactose due to intramolecular steric interactions. The most potent E-selectin antagonist 6v showed >50-fold improved E-selectin inhibition compared to the weak selectin ligand sialyl Lewis' (sLe^x, 1: IC₅₀ = 1000-1500_μM), but only a 2-fold improvement compared to 3. Compound 6x was tested in vivo in a murine model of acute inflammation and found to be as potent as 3 (ED₅₀ = 15 mg/kg).

Full text of DOI:10.1002/1522-2675(20001108)83:11<2893::AID-HLCA2893>3.0.CO;2-G

Helvetica Chimica Acta ± Vol. 83 02000)
2893
Potent E-Selectin Antagonists
by Rolf Bäntelia), Peter Herolda), Christian Brunsa), John T. Pattonb), John L. Magnanib),
and Gebhard Thoma*a)1)
a) Novartis Pharma AG, P.O. Box, CH-4002 Basel
b) GlycoTech Corporation, 14915 Broschart Road, Rockville, Maryland 10850, USA
In the search for drugs that could control excessive leukocyte extravasation, we now report on
modifications of the already known potent E-selectin antagonist 3 containing a cyclohexyllactic acid residue and
a glucal-derived building block. Thus, we describe the synthesis and biological evaluation of a series of
derivatives 6 with modified glucal-derived moieties 0CH2NR1R2 instead of CH2OH in 3) to explore a
hypothetical potential complementary interaction with E-selectin. However, similar activity profiles of most
derivatives 6 and compound 3 do not support such an interaction, but rather indicate topological-structure
changes of 6 0and 3) in the orientation of the neighboring fucose and galactose due to intramolecular steric
interactions. The most potent E-selectin antagonist 6v showed >50-fold improved E-selectin inhibition
compared to the weak selectin ligand sialyl Lewisx 0sLex, 1; IC50 ˆ 1000 ± 1500 mm), but only a 2-fold
improvement compared to 3. Compound 6x was tested in vivo in a murine model of acute inflammation and
found to be as potent as 3 0ED50 ˆ 15 mg/kg).
Introduction. ± Excessive infiltration of leukocytes from blood vessels into
surrounding tissues, mediated by cell-adhesion molecules 0CAM) and extracellular
matrix proteins, can cause both acute and chronic inflammatory disorders such as
reperfusion injuries, psoriasis, rheumatoid arthritis, or respiratory diseases [1]. E- and
P-selectin ± prominent members of the CAM family ± that are expressed on endothelial
cells upon stimulation, are involved in an early step of the cascade of events that finally
leads to the extravasation of leukocytes [2]. E-Selectin recognizes complex glyco-
proteins on the leukocyte surface by interacting with the carbohydrate part of the
physiological ligand [3]. For P-selectin, additional interactions with sulfated tyrosines
have been discussed [4]. It has been indicated that the tetrasaccharide sialyl Lewisx
0sLex; 1) is an epitope of the physiological selectin ligands and is a weak selectin
inhibitor itself 0E-selectin, IC50 ˆ 1000 ± 1500 mm) [5]. Simplified, more-potent analogs
of the complex tetrasaccharide 1 could be used to control inflammatory disorders and,
therefore, are of pharmaceutical interest.
Extensive work elucidated that the essential pharmacophores of sLex required to
bind to E-selectin are the carboxylic acid function, all three OH groups of the fucose,
and the 4-OH and 6-OH groups of galactose. Based on this knowledge, a multitude of
small-molecule selectin antagonists have been designed during the last few years. The
efforts of the various research groups involved have been summarized in an excellent
review [6]. The E-selectin antagonist 2 with 10-fold improved activity compared to sLex
01) was obtained by concomitant replacement of sialic acid by cyclohexyllactic acid and
1
)
E-mail: gebhard.thoma@pharma.novartis.com

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Helvetica Chimica Acta ± Vol. 83 02000)
of N-acetylglucosamine by cyclohexanediol [7]. Upon incorporation of a glucal-
derived building block instead of the expensive chiral cyclohexanediol, we discovered
compound 3, which was even more potent than 2 03-fold improvement in a static assay;
6-fold improvement in a dynamic flow assay) [8]. Possible explanations are additional
direct interactions with E-selectin or conformational changes of 3 compared to 2.
Interestingly, the regioisomer 4 of antagonist 3 did not show improved potency as
compared to 2 [9].
Recently, Ramphal et al. have reported on N-acylglucosamine derivatives 5 of sLex
[10]. Aromatic residues such as 2-naphthyl in 5a and phenyl in 5b led to 10- and 3-fold
improved affinities, respectively, compared to sLex, whereas the corresponding
cyclohexyl derivative 5c was less active than sLex. Since these compounds showed
essentially the same topological-structure as sLex, the effect was attributed to an
additional interaction between aromatic residues and a new binding site on E-selectin
[10]. Current models of ligand binding on E-selectin do not account for a
complementary lipophilic binding site but, due to the lack of an X-ray crystal structure
of a ligand/E-selectin complex, these models are mainly based on indirect evidence
obtained from the X-ray structure of the lectin domain of E-selectin, molecular
modeling, and NMR studies [11].
It has been shown that the bound conformations of sLex and antagonist 2 on E-
selectin are very similar [11c, d]. Assuming related bioactive conformations of 3 and 2
and of sLex and 5, we concluded that the CH2OH group of the glucal-derived moiety of
3 and the aromatic residues of sLex derivatives 5 point in the same direction and address
the same area of the protein. Thus, suitable substituents at this position of 3 could lead

Helvetica Chimica Acta ± Vol. 83 02000)
2895
to compounds with improved affinity. Since 3 seems to be more potent than 5a2),
derivatives of 3 could be expected to be highly active E-selectin antagonists. To explore
the hypothesis of a potential complementary binding site of E-selectin, we decided to
prepare a series of compounds 6 with various substituents in the glucal-derived portion.
In this paper, we describe the synthesis of antagonists 6 and their biological
evaluation in both static and dynamic E-selectin in vitro assays. Most compounds 6
showed a similar activity profile to that of 3. The best antagonist 6v exhibited a >50-
fold improved E-selectin inhibition compared to sLex 01), but only a 2-fold improve-
ment compared to 3. These findings do not support the hypothesis of a complementary
hydrophobic binding site of E-selectin, but rather indicate topolocial-structure changes
in the orientation of the neighboring fucose and galactose due to intramolecular steric
interactions in 3 and 6.
Synthesis. ± To modify the glucal-derived portion of compound 3 in a convergent
manner, we required a selectively protected advanced intermediate with a free primary
OH group, such as 7. Thus, the previously developed synthesis that readily gave access
to antagonist 3 could not be applied [8]. The newly developed synthesis is depicted in
the Scheme. Compound 7 was prepared from thiogalactoside 8 [12], cyclohexyllactic
acid derivative 9 [8], glucal-derived building block 10 [8], and thiofucoside 11 [13]
0Scheme). Tin-mediated alkylation of 8 with 9, followed by benzoylation of
intermediate 12, gave the modified galactose donor 13. Glycosylation of acceptor 10
and subsequent removal of the benzylidene acetal afforded disaccharide 14. Surpris-
ingly, in addition to the desired product 14, we isolated 10 ± 15% of the corresponding
a-d-anomer 15. This was unexpected since the neighboring-group participation of the
benzoate in the 2-position of donor 13 should selectively lead to the b-d-glycosidic
linkage. The stereoisomers were separated by chromatography. Next, the primary OH
group of 14 was silyl-protected to give 16, which was fucosylated using thioglycoside 11.
Donor 11 was transformed into the corresponding glycosyl bromide, which gave, under
in situ anomerization conditions in the presence of Et4NBr, exclusively the a-l-
fucoside. Desilylation of the crude material with Bu4NF led to partial cleavage of the
sensitive a-l-glycosidic bond between fucose and galactose, but treatment with H2SiF6
[14] furnished compound 7 in 80% yield over 2 steps. Comparable yields were obtained
using a- or b-l-thiofucoside 11. The selectively protected, advanced intermediate 7 was
readily available in multi-gram quantities.
The unprotected primary OH group of 7 was converted to an amino function by
preparing mesylate 17, which was subsequently transformed into azide 18. Saponifi-
cation gave 19 which was completely deprotected by subsequent hydrogenation to
furnish amino acid 6a. Compound 6a is available from thiogalactoside 8 in 12 steps with
an overall yield of 20%. Several analogs such as amines 6b and 6c, derivatives with
negatively charged residues 6d and 6e, amides with aromatic residues 6f ± p, aliphatic
amides 6q and 6r, sulfonamide 6s, ureas 6t and 6u, and carbamates 6v ± x were prepared
from 6a. The yields of 6b ± x are given in the Table, and the conditions for the individual
transformations are described in detail in the Exper. Part.
2
)
It is worth noting that compounds 5 and 3 have been tested in different assays. Comparison of affinities
could be misleading, even though sLex was used as a reference in both assay formats.

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Scheme
a) Bu2SnO, MeOH, D. b) 9, CsF, MeOCH2CH2OMe. c) BzCl, N,N-dimethylpyridin-4-amine 0DMAP), Py. d) 10
t
N-iodosuccinimide 0NIS), TfOH, CH2Cl2. e) AcOH, H2O, D. f) BuMe2SiCl, Py. g) 11, Br2, Et4NBr, CH2Cl2/
DMF. h) H2SiF6Et3N, MeCN. i) MeSO2Cl, Py. j) NaN3, DMF. k) LiOH, H2O, MeOH. l) H2, 10% Pd/C, dioxane,
H2O.
Biological Evaluation. ± All compounds 6 were tested at least twice in a
competitive, cell-free assay that measures E-selectin inhibition under equilibrium
conditions [15]. E-Selectin/hIg chimera is immobilized on microtiter plates and
incubated with a biotinylated polylysine-sLea conjugate. The displacement of this
multivalent ligand by E-selectin inhibitors was monitored and quantified to determine
their IC50 values 0concentration to achieve 50% displacement of the polymer). To
compare the data for different compounds obtained on different test plates, sLex 01)
was assayed on each plate as a reference. This allows the determination of IC50 values
relative to sLex, which are defined as rel. IC50 ˆ IC500test compound)/IC500sLex).
Since E-selectin-mediated rolling of leucocytes on activated endothelium is
expected to be a nonequilibrium process, we also used a cell-based flow assay to

Helvetica Chimica Acta ± Vol. 83 02000)
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Table. Compositon and E-Selectin Inhibition Data ofCompounds 3 and 6a ± x
6-Substituent of the
glucal-derived moiety
Yield [%] Static assay Flow assay
a
rel. IC50
)
[% red. of NIC]b) at 50 mm 0rel. red. NIC)c)
3
OH
0.032
0.404
0.693
0.520
0.046
0.032
0.034
0.031
0.031
0.057
0.065
0.024
0.028
0.075
0.084
0.094
0.028
0.038
0.044
0.039
0.053
0.045
0.017
0.021
0.034
60% 01.36)
n.d.
n.d.
6a NH2
6b NHCH2Ph
6c N0CH2Ph)2
6d NHSO3Na
78
14
36
46
27
40
55
40
51
46
n.d.
36% 00.68)
66% 01.30)
n.d.
78% 01.03)
85% 01.12)
78% 01.04)
70% 01.43)
62% 00.84)
70% 01.00)
n.d.
6e NHCO-C6H4ÀCOOH 0ortho)
6f NHCOPh
6g NHCOÀC6H4ÀCl 0para)
6h NHCOÀC6H4ÀOMe 0para)
6i NHCOÀC6H4ÀNO2 0para)
6j NHCOÀC6H4ÀPh 0para)
6k NHCOÀC6H30OMe)2 0meta, para) 82
6l NHCO02-naphthyl)
6m NHCOÀC6H4ÀOCH2Ph 0para)
6n N0CH2Ph)COPh
6o NHCOCH2CH2Ph
6p NHCOCHPh2
78
79
48
49
79
46
89
42
64
42
n.d.
n.d.
70% 00.95)
41% 00.57)
38% 00.71)
74% 01.20)
51% 00.82)
68% 00.88)
87% 00.98)
92% 01.03)
82% 01.06)
6q NHCOMe
6r NHCO0cyclo-C6H11)
6s NHSO2ÀC6H4ÀMe 0para)
6t NHCONHEt
6u NHCONHPh
6v NHCOOCH2ÀC6H4ÀNO2 0para) 50
6w NHCOOCH202-naphthyl)
6x NHCOOCH2Ph
30
72
a) Relative IC50 values are defined as rel. IC50 ˆ IC500test compound)/IC500sLex); sLex 01) was tested as a
reference on each plate to allow direct comparison of data from different test plates; compounds were tested at
least twice 0mean value). b) Reduction of the number of interacting cells 0NIC). c) Relative reductions of the
number of interacting cells 0NIC) are defined as rel. red. NIC ˆ red. 0compound at 50 mm)/reduction 02 at
200 mm). On each test day, 2 was tested as a reference to be able to compare data from individual test days and
different cell cultures.
characterize antagonists 6 0Fig.) [16]. This dynamic in vitro assay monitors the rolling
of polymorphonuclear neutrophils 0PMN) on stimulated human umbilical vein
endothelium cells 0HUVEC) under hydrodynamic shear stress and, thus, mimics in
vivo conditions. The reduction of the number of interacting cells 0NIC) serves as a
measure to determine the inhibitory potential of a test compound. The assay is labor-
intensive, and only 4 ± 5 compounds can be tested on a single day. New HUVECs from
different sources are used on each test day. This can affect the E-selectin expression
levels and, consequently, causes different assay sensitivities on individual test days. To
allow a more accurate comparison of the data obtained on individual test days,
compound 2 was used as a reference and assayed on each test day at 200 mm along with
the test compounds. The test compounds were assayed at 50 mm. The relative reduction
of the number of interacting cells 0rel. red. NIC) of an antagonist with respect to 2 is
defined as rel. red. NIC ˆ red. NIC0test compound at 50 mm)/red. NIC02 at 200 mm).
The dynamic cell assay was performed for the compounds that showed promising

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affinities in the primary, static assay. The in vitro data for E-selectin inhibitors 6a ± x are
given in the Table and are compared to 3.
Figure. Cell-based E-selectin flow assay
The amines 6a ± c showed similar activities in the static assay 0rel. IC50 ˆ 0.4 ± 0.7),
despite of their different substitution patterns. They were more potent than sLex 02-
fold), but significantly less active than 3 010-fold). Unfavorable steric and/or electronic
interactions of the substituents with E-selectin are not probable since, in case of direct
contact with the protein, the different steric demands of the substituents should be
reflected by less-uniform bioactivities. It is more likely that intramolecular electrostatic
interactions between the amine and the carboxylic acid unfavorably affect the
conformations of these antagonists. The dependence of the potency of E-selectin
antagonists on their conformation in solution has been outlined earlier [7]. Due to their
relatively weak affinities, these compounds have not been tested in the flow assay.
Interestingly, analogs with an additional negatively charged substituent, such as 6d and
6e 0rel. IC50 ˆ 0.046 and 0.032, resp.), were much more potent than the amines but did
not exhibit improved affinities compared to 3 0rel. IC50 ˆ 0.032). In the flow assay,
antagonist 6e was as potent as 3, whereas 6d was less active.
The amides 6f ± p with various aromatic substituents showed good affinities to E-
selectin within a relatively narrow range 0rel. IC50 ˆ 0.024 ± 0.094), but compared to 3
0rel. IC50 ˆ 0.032), no substantial improvement could be achieved. Surprisingly, the
aliphatic amides 6q and 6r were found to be as active as the aromatic derivatives.
Comparable affinities were observed for naphthalene-2-carboxamide 6l 0rel. IC50 ˆ
0.028) and cyclohexanecarboxamide 6r 0rel. IC50 ˆ 0.044), contrary to the results for
the corresponding sLex derivatives 5. In the flow assay, the aliphatic amides 6q and 6r
were found to be slightly less-potent than 3, whereas the compounds with aromatic
substituents showed very similar activities. Our findings seem to contradict the results
reported by Ramphal et al., but it has to be considered that we investigated
substantially modified sLex mimics that could behave differently than the N-acyl-
substituted sLex pentasaccharides 5. For example, in 6, the amide function is separated
from the six-membered ring by a CH2 group inducing a higher degree of flexibility than
in 5 where it is directly attached to the ring.
The aromatic sulfonamide 6s 0rel. IC50 ˆ 0.039), and the ureas 6t and 6u 0rel. IC50 ˆ
0.045 ± 0.053) were found to be similarly as potent as 3. The most-active compounds
obtained were among the carbamates. Antagonist 6v showed 2-fold-improved potency

Helvetica Chimica Acta ± Vol. 83 02000)
2899
0rel. IC50 ˆ 0.017) compared to 3, while 6w 0rel. IC50 ˆ 0.021) was slightly more active
than 3, and 6x was equally active 0rel. IC50 ˆ 0.034). Generally, the data from the static
assay could be confirmed by the flow assay.
The in vivo efficacy of carbamate 6x was tested in a murine peritonitis model and
compared to the more hydrophilic compound 3 [17]. The migration of leukocytes in
response to an acute inflammatory stimulus was assessed by intraperitoneal injection of
thioglycollate followed by an appropriate incubation. Peritonitis was induced in female
NMRI mice 022 ± 25 g) by intraperitoneal 0i.p.) injection of 1 ml of 3% thioglycolate in
0.9% 0w/v) NaCl solution. Negative-control mice received an injection of sterile saline
only. The total number of PMNs in the peritoneal exudates increased 13-fold within 3 h
of thioglycolate treatment. Animals were killed 3 h post injection and the peritoneal
cavity lavaged with 5 ml of ice-cold PBS 0containing 188 U/ml of heparin). Total cell
counts were performed and cytospins prepared for differential cell counting to
determine the number of neutrophils transmigrated into the peritoneum. Neutrophil
infiltration of the inflamed peritoneum was dose-dependently inhibited by the anti-E-
selectin antibody 10E9.6, resulting in a complete inhibition of the thioglycolate-induced
peritonitis at 30 mg/mouse 0ED50 ˆ 5 mg/mouse). In contrast, injection of a negative IgG
control antibody 010 mg/mouse) did not alter thioglycolate-induced neutrophil influx.
The demonstration that thioglycolate-induced cell influx can be inhibited by a specific
anti-E-selectin antibody provides proof-of-concept that the peritonitis model can be
used to test small-molecular-weight selectin inhibitors. Test compounds were admin-
istered i.p. at the time of thioglycolate stimulation. Compound 3 showed promising
activity in this model earlier 0ED50 ˆ 15 mg/kg) [8]. Compound 6x was found to inhibit
the transmigration of PMNs into the peritoneum during thioglycolate-induced acute
inflammation with equal efficacy 0ED50 ˆ 15 mg/kg), whereas sLex was ineffective at
doses up to 100 mg/kg.
Conclusion. ± Similar affinity profiles of E-selectin antagonist 3 and a series of
derivatives 6 with modifications in the glucal-derived moiety do not support the
hypothesis of an additional interaction of the substituent with a complementary
lipophilic binding site. It is more likely that a steric interaction of the CH2R substituent
of compounds 3 and 6 with the neighboring fucose may affect the solution
conformation of these compounds, as suggested by a strong NOE between the CH2
protons of the CH2R substituent and HÀC01) of the fucose. We are currently
investigating this hypothesis. Antagonist 6v showed >50-fold improved potency
compared to sLex, but only 2-fold improved potency compared to compound 3. A
variety of substituents are tolerated without losing affinity, allowing for the
modification of pharmacokinetic properties.
Experimental Part
General. All reactions were carried out under dry Ar. Commercially available abs. solvents were used.
Column chromatography ˆ CC, flash chromatography ˆ FC. NMR Spectra: Bruker-Avance-DPX-400 spec-
trometer; assignments by 2-D 1H,1H correlation 0COSY) and 1H,13C correlation 0HSQC); d in ppm rel. to
SiMe4, J in Hz; Chx ˆ cyclohexyl, Hex ˆ arabino-d-hexitol. MS: Finnigan-MAT-90 mass spectrometer.
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-6-amino-1,5-anhydro-2,6-dideoxy-d-arabino-hexitol 06a). To a soln. of 19 08.45 g, 9.33 mmol) in
dioxane 0250 ml) and H2O 050 ml), 10% Pd/C 03.4 g) was added. The suspension was stirred vigorously for 20 h

2900
Helvetica Chimica Acta ± Vol. 83 02000)
under H2 and then filtered through Celite ꢀ 0washing with dioxane). The pH of the filtrate was brought from 6 to
5 by adding AcOH 04 ml). After addition of fresh catalyst 010% Pd/C 03.4 g) suspended in H2O 020 ml)),
hydrogenation was continued for 20 h. Filtration 0Celiteꢀ), evaporation and CC 0silica gel 0300 g), AcOEt/
iPrOH/H2O 2 :2 :1) yielded 3.82 g 067%) of 6a. 1H-NMR 0400 MHz, D2O): 0.7 ± 0.95 0m, 2 H); 0.95 ± 1.2
0m, 3 H) and 1.35 ± 1.75 0m, 9 H) 0HaxÀC02)0Hex), CH20Chx)); 1.10 0d, J ˆ 6.5, Me06)0Fuc)); 2.07 ± 2.2 0m,
HeqÀC02)0Hex)); 3.0 ± 3.15 0m, 1 HÀC06)0Hex)); 3.29 0dd, J ˆ 10.0, 3.5, HÀC03)0Gal)); 3.3 ± 3.6
0m, 1 HÀC01)0Hex), HÀC04)0Hex), HÀC05)0Hex), 1 HÀC06)0Hex), HÀC02)0Gal), HÀC05)0Gal)); 3.6 ±
3.65 0m, 2 HÀC06)0Gal)); 3.65 ± 3.73 0m, HÀC02)0Fuc), HÀC04)0Fuc)3)); 3.73 ± 3.83 0m, HÀC04)0Gal),
HÀC03)0Fuc)); 3.83 ± 3.9 0m, OCHCO2H); 3.9 ± 4.02 0m, 1 HÀC01)0Hex), HÀC03)0Hex)); 4.42 0d, J ˆ 8.0,
HÀC01)0Gal)); 4.58 0q, J ˆ 6.5, HÀC05)0Fuc)); 4.80 0d, J ˆ 3.5, HÀC01)0Fuc)). HR-MS: 610.3079
‡
‡
0C27H48NO14 , [M ‡ H] ; calc. 610.3075).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-2,6-dideoxy-6-[0phenylmethyl)amino]-d-arabino-hexitol 06b) and O-3-O-[01S)-1-Car-
boxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyranosyl-01 ! 4)]-1,5-anhydro-
6-[bis0phenylmethyl)amino]-2,6-dideoxy-d-arabino-hexitol 06c). To a mixture of 6a 040 mg, 0.066 mmol),
benzaldehyde 00.033 ml, 0.328 mmol), and freshly dried 4-Š molecular sieves 0ca. 500 mg) in dry MeOH
00.5 ml), borane-pyridine complex 0BH3 ´ C5H5N; 0.013 ml, 0.131 mmol) was added. After 20 h, the mixture 02
new products) was filtered and evaporated and the residue separated by FC 0silica gel, AcOEt/iPrOH/H2O
4 :2.1): 6c and then 6b. Each was separately further purified by gel filtration 0P2, H2O) and by ion-exchange
‡
chromatography 0Dowex 0Na form), H2O) and then freeze-dried: 6c 017 mg, 36%) and 6b 06.4 mg, 14%),
resp., both as white foams.
Data of 6b: 1H-NMR 0400 MHz, D2O): 0.72 ± 0.92 0m, 2 H); 0.95 ± 1.21 0m, 6 H, including a d at 1.09, J ˆ 7,
Me06)0Fuc)); 1.38 ± 1.72 0m, 8 H); 1.65 ± 1.73 0m, 1 H); 2.08 ± 2.18 0m, 1 H); 3.18 0dd, J ˆ 10, 12, 1 H); 3.29
0dd, J ˆ 10, 3, HÀC03)0Gal)); 3.32 ± 3.44 0m, 2 H); 3.45 ± 3.59 0m, 4 H); 3.59 ± 3.68 0m, 3 H); 3.69 0d, 1 H); 3.73
0dd, J ˆ 3, 11, HÀC03)0Fuc)); 3.81 0d, J ˆ 3, HÀC04)0Gal)); 3.48 ± 4.00 0m, 3 H); 4.14 0d, J ˆ 12, 1 H, PhCH2);
4.19 0d, J ˆ 12, 1 H, PhCH2); 4.41 0d, J ˆ 8, HÀC01)0Gal)); 4.52 0q, J ˆ 7, H ÀC05)0Fuc)); 4.72 0d, J ˆ 4,
HÀC01)0Fuc)); 7.38 0m, 5 H). HR-MS: 698.3380 0C34H52NO14À, [M À Na]À; calc. 698.3388).
Data of 6c: The 1H-NMR showed broad signals that were shifted in different measurements; a total of 2 H
were missing in the region 3.0 ± 4.7 ppm, probably due to signal broadening. 1H-NMR 0400 MHz, D2O): 0.75 ±
0.92 0m, 2 H); 0.97 ± 1.22 0m, 6 H, including a d at 1.08, J ˆ 7, Me06)0Fuc)); 1.38 ± 1.73 0m, 8 H); 1.65 ± 1.74
0m, 1 H); 2.02 ± 2.12 0m, 1 H); 3.19 ± 3.37 0m, 3 H); 3.37 ± 3.58 0m, 3 H); 3.58 ± 3.78 0m, 7 H); 3.81 0d, J ˆ 3,
HÀC04)0Gal)); 3.83 ± 3.93 0m, 2 H); 4.12 ± 4.46 0m, 3 H); 4.37 0d, J ˆ 8, HÀC01)0Gal)); 4.43 0q, J ˆ 7,
HÀC05)0Fuc)); 4.64 0d, J ˆ 4, HÀC01)0Fuc)); 7.41 0m, 10 H). HR-MS: 788.3862 0C41H58NO14À, [M À Na]À;
calc. 788.3857).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-2,6-dideoxy-6-0sulfoamino)-d-arabino-hexitol Sodium Salt 06d). To a soln. of 6a
020 mg, 0.033 mmol) in H2O 02 ml) with enough 2n NaOH to obtain a pH >11, commercially available sulfur
trioxide pyridine complex 07.8 mg, 0.049 mmol, 1.5 equiv.) was added. After 16 h, another portion of sulfur
trioxide pyridine complex 07.8 mg, 0.049 mmol, 1.5 equiv.) was added. After 40 h, the mixture was evaporated
and the residue purified by gel filtration 0P2, BioRad, H2O) and CC 0RP C18, SepPack syringe adapter, step
gradient MeCN/H2O 1:9, 2 :8, 3 :7, 4 :6, 5 :5, 7:3, and 9 :1) and freeze-dried: 6d 010.6 mg, 46%). White foam.
1H-NMR 0400 MHz, D2O): 0.75 ± 0.92 0m, 2 H); 1.00 ± 1.21 0m, 6 H, including a d at 1.12, J ˆ 7, Me06)0Fuc));
1.39 ± 1.63 0m, 8 H); 1.66 ± 1.74 0m, 1 H); 2.08 ± 2.17 0m, 1 H); 3.12 0dd, J ˆ 7, 12, 1 H); 3.30 0dd, J ˆ 10, 3,
HÀC03)0Gal)); 3.32 ± 3.48 0m, 4 H); 3.48 ± 3.58 0m, 2 H); 3.59 ± 3.67 0m, 2 H); 3.67 ± 3.73 0m, HÀC04)0Fuc),
HÀC02)0Fuc)); 3.78 0dd, J ˆ 3, 11, HÀC03)0Fuc)); 3.82 0d, J ˆ 3, HÀC04)0Gal)); 3.86 ± 3.99 0m, 3 H); 4.42
0d, J ˆ 8, HÀC01)0Gal)); 4.68 0q, J ˆ 7, H ÀC05)0Fuc)); 4.98 0d, J ˆ 4, HÀC01)0Fuc)). HR-MS: 688.2491
0C27H46NO17SÀ, [M À 2 Na ‡ H]À; calc. 688.2486).
General Procedure 0G.P.) for the Synthesis of Amides 6e ± r and Carbamates 6v ± x. To a soln. of 6a 020 mg,
0.033 mmol) in THF/H2O 1:1 02 ml) at 08, a soln. of commercially available acid chloride 00.049 mmol,
1.5 equiv.) in THF 00.5 ml) was added. The pH of the mixture was adjusted to 8 ± 10 by the addition of 1n NaOH
and maintained at 8 ± 10 throughout the reaction. If necessary, additional acid chloride 00.016 mmol, 0.5 equiv.)
was added after 1 ± 4 h, and after a total of 2 ± 42 h, the mixture was partially evaporated to remove THF. The
now aq. soln. was purified by applying one or more of the following three purification methods, as needed, to get
3
)
6-Deoxy-a-l-galactopyranose refers to a-l-fucopyranose 0Fuc).

Helvetica Chimica Acta ± Vol. 83 02000)
2901
a pure product: FC 0silica gel, AcOEt/iPrOH/H2O 4 :2 :1) and/or CC 0RP C18, column 1 Â 10 cm, MeCN/H2O
3 :7, then MeCN/H2O 4 :6), and/or gel filtration 0P2, BioRad, H2O). The product obtained was freeze dried:
white foam.
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-6-[02-carboxybenzoyl)amino]-2,6-dideoxy-d-arabino-hexitol 06e). According to the
G.P. FC, gel filtration, and CC gave 10.3 mg 027%) of 6e. 1H-NMR 0400 MHz, D2O): 0.75 ± 0.90 0m, 2 H); 1.00 ±
1.2 0m, 6 H, including a d at 1.12, J ˆ 7, Me06)0Fuc)); 1.39 ± 1.65 0m, 8 H); 1.65 ± 1.73 0m, 1 H); 2.10 ± 2.18
0m, 1 H); 3.30 0dd, J ˆ 10, 3, HÀC03)0Gal)); 3.36 ± 3.58 0m, 6 H); 3.58 ± 3.65 0m, 2 H); 3.68 ± 3.75
0m, HÀC04)0Fuc), HÀC02)0Fuc)); 3.75 ± 3.83 0m, 3 H); 3.83 ± 4.04 0m, 3 H); 4.42 0d, J ˆ 8, HÀC01)0Gal));
4.64 0q, J ˆ 7, H ÀC05)0Fuc)); 4.91 0d, J ˆ 4, HÀC01)0Fuc)); 7.35 ± 7.44 0m, 3 H); 7.49 0d, J ˆ 7, 1 H). HR-MS:
756.3080 0C35H50NO17À, [M À Na]À; calc. 756.3079).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-6-0benzoylamino)-2,6-dideoxy-d-arabino-hexitol 06f). According to the G.P. FC
followed by gel filtration gave 9.7 mg 040%) of 6f. 1H-NMR 0400 MHz, D2O): 0.73 ± 0.92 0m, 2 H); 1.00 ± 1.21
0m, 6 H, including a d at 1.13, J ˆ 7, Me06)0Fuc)); 1.39 ± 1.65 0m, 8 H); 1.65 ± 1.73 0m, 1 H); 2.10 ± 2.18 0m, 1 H);
3.32 0dd, J ˆ 10, 3, HÀC03)0Gal)); 3.36 ± 3.69 0m, 8 H); 3.68 ± 3.75 0m, HÀC04)0Fuc), HÀC02)0Fuc)); 3.80
0dd, J ˆ 3, 11, HÀC03)0Fuc)); 3.75 ± 3.88 0m, 2 H, including a d at 3.85, J ˆ 3, HÀC04)0Gal)); 3.90 ± 4.04
0m, 3 H); 4.42 0d, J ˆ 8, HÀC01)0Gal)); 4.64 0q, J ˆ 7, H ÀC05)0Fuc)); 4.93 0d, J ˆ 4, HÀC01)0Fuc)); 7.42 0t, J ˆ
7, 2 H); 7.52 0m, 1 H); 7.66 0d, J ˆ 7, 2 H). HR-MS: 712.3178 0C34H50NO15À, [M À Na]À, calc. 712.3180).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-6-[04-chlorobenzoyl)amino]-2,6-dideoxy-d-arabino-hexitol 06g). According to the
G.P. FC gave 13.8 mg 055%) of 6g. 1H-NMR 0400 MHz, D2O): 0.72 ± 0.92 0m, 2 H); 1.00 ± 1.21 0m, 6 H,
including a d at 1.10, J ˆ 7, Me06)0Fuc)); 1.37 ± 1.65 0m, 8 H); 1.65 ± 1.73 0m, 1 H); 2.10 ± 2.18 0m, 1 H); 3.30
0dd, J ˆ 10, 3, HÀC03)0Gal)); 3.35 ± 3.68 0m, 8 H); 3.68 ± 3.73 0m, HÀC04)0Fuc), HÀC02)0Fuc)); 3.78 0dd, J ˆ
3, 11, HÀC03)0Fuc)); 3.75 ± 3.83 0m, 2 H, including a d at 3.81, J ˆ 3, HÀC04)0Gal)); 3.84 ± 4.04 0m, 3 H); 4.42
0d, J ˆ 8, HÀC01)0Gal)); 4.63 0q, J ˆ 7, H ÀC05)0Fuc)); 4.92 0d, J ˆ 4, HÀC01)0Fuc)); 7.42 0m, 2 H); 7.62
0m, 2 H). HR-MS: 746.2789 0C34H49ClNO15À, [M À Na]À; calc. 746.2791).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-2,6-dideoxy-6-[04-methoxybenzoyl)amino]-d-arabino-hexitol 06h). According to the
G.P. FC gave 10.3 mg 040%) of 6h.1H-NMR 0400 MHz, D2O): 0.75 ± 0.92 0m, 2 H); 1.00 ± 1.21 0m, 6 H, including
a d at 1.10, J ˆ 7, H ÀC06)0Fuc)); 1.39 ± 1.65 0m, 8 H); 1.65 ± 1.73 0m, 1 H); 2.11 ± 2.18 0m, 1 H); 3.30 0dd, J ˆ 10,
3, HÀC03)0Gal)); 3.35 ± 3.68 0m, 8 H); 3.68 ± 3.73 0m, HÀC04)0Fuc), HÀC02)0Fuc)); 3.78 0dd, J ˆ 3, 11,
HÀC03)0Fuc)); 3.79 0s, MeO); 3.75 ± 3.83 0m, 2 H, including a d at 3.81, J ˆ 3, HÀC04)0Gal)); 3.85 ± 4.04
0m, 3 H); 4.42 0d, J ˆ 8, HÀC01)0Gal)); 4.63 0q, J ˆ 7, H ÀC05)0Fuc)); 4.92 0d, J ˆ 4, HÀC01)0Fuc)); 6.98
0m, 2 H); 7.66 0m, 2 H). HR-MS: 742.3282 0C35H52NNaOÀ16, [M À Na]À; calc. 742.3286).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-2,6-dideoxy-6-[04-nitrobenzoyl)amino]-d-arabino-hexitol 06i). According to the G.P.
FC gave 13 mg 051%) of 6i. 1H-NMR 0400 MHz, D2O): 0.75 ± 0.90 0m, 2 H); 0.96 ± 1.21 0m, 6 H, including a d at
1.10, J ˆ 7, Me06)0Fuc)); 1.36 ± 1.64 0m, 8 H); 1.64 ± 1.72 0m, 1 H); 2.08 ± 2.18 0m, 1 H); 3.29 0dd, J ˆ 10, 3,
HÀC03)0Gal)); 3.35 ± 3.68 0m, 5 H); 3.58 ± 3.74 0m, 5 H); 3.76 ± 3.83 0m, 2 H, HÀC04)0Gal)); 3.84 ± 4.04
0m, 4 H); 4.42 0d, J ˆ 8, HÀC01)0Gal)); 4.63 0q, J ˆ 7, H ÀC05)0Fuc)); 4.92 0d, J ˆ 4, HÀC01)0Fuc)); 7.83
0m, 2 H); 8.25 0m, 2 H). HR-MS: 757.3026 0C34H49N2O17À, [M À Na]À; calc. 757.3031).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-6-0[1,1'-biphenyl]-4-ylamino)-2,6-dideoxy-d-arabino-hexitol 06j). According to the
G.P. FC followed by gel filtration gave 12 mg 046%) of 6j. 1H-NMR 0400 MHz, D2O): 0.78 ± 0.95 0m, 2 H);
1.00 ± 1.22 0m, 6 H, including a d at 1.13, J ˆ 7, Me06)0Fuc)); 1.40 ± 1.68 0m, 8 H); 1.68 ± 1.76 0m, 1 H); 2.13 ± 2.21
0m, 1 H); 3.32 0dd, J ˆ 10, 3, HÀC03)0Gal)); 3.38 ± 3.62 0m, 5 H); 3.62 ± 3.72 0m, 3 H); 3.72 ± 3.80
0m, HÀC04)0Fuc), HÀC02)0Fuc)); 3.80 ± 4.08 0m, 6 H); 4.45 0d, J ˆ 8, HÀC01)0Gal)); 4.66 0q, J ˆ 7,
HÀC05)0Fuc)); 4.97 0d, J ˆ 4, HÀC01)0Fuc)); 7.42 0m, 1 H); 7.48 0t, J ˆ 7, 2 H); 7.65 ± 7.75 0m, 4 H); 7.78
‡
‡
0d, J ˆ 7, 2 H). HR-MS: 812.3471 0C40H55NNaO15 , [M ‡ H] ; calc. 812.3469).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-6-[03,4-dimethoxybenzoyl)amino]-2,6-dideoxy-d-arabino-hexitol 06k). According to
the G.P. CC followed by FC gave 21.5 mg 082%) of 6k. 1H-NMR 0500 MHz, D2O): 0.82 ± 0.96 0m, 2 H); 1.08 ±
1.24 0m, 6 H, including a d at 1.17, J ˆ 6.5, Me06)0Fuc)); 1.45 ± 1.69 0m, 8 H); 1.72 ± 1.78 0m, 1 H); 2.19 ± 2.24
0m, 1 H); 3.36 0dd, J ˆ 9.3, 3.1, HÀC03)0Gal)); 3.44 ± 3.54 0m, 2 H); 3.54 ± 3.74 0m, 6 H); 3.75 ± 3.80
0m, HÀC04)0Fuc), HÀC02)0Fuc)); 3.75 0dd, J ˆ 9.5, 3.0, HÀC03)0Fuc)); 3.81 ± 3.90 0m, 8 H, including a s at

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Helvetica Chimica Acta ± Vol. 83 02000)
3.86, MeO); 3.87 0s, MeO); 3.93 0m, 1 H); 3.99 0m, 1 H); 4.05 0m, 1 H); 4.49 0d, J ˆ 7.8, HÀC01)0Gal)); 4.69
0q, J ˆ 6.8, HÀC05)0Fuc)); 4.98 0d, J ˆ 3.7, HÀC01)0Fuc)); 7.06 0d, J ˆ 8.6, 1 H); 7.35 0d, J ˆ 2.2, 1 H); 7.40
‡
‡
0dd, J ˆ 2.2, 8.6, 1 H). HR-MS: 0C36H55NNaO17 , [M ‡ H] ; calc. 796.3368).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-2,6-dideoxy-6-[0naphthalen-2-ylcarbonyl)amino]-d-arabino-hexitol 06l). According to
the G.P. CC followed by FC gave 20.4 mg 078%) of 6l. 1H-NMR 0500 MHz, D2O): 0.81 ± 1.00 0m, 2 H); 1.06 ±
1.24 0m, 6 H, including a d at 1.10, J ˆ 6.3, Me06)0Fuc)); 1.46 ± 1.72 0m, 8 H); 1.72 ± 1.79 0m, 1 H); 2.18 ± 2.24
0m, 1 H); 3.36 0dd, J ˆ 9.5, 3.3, HÀC03)0Gal)); 3.45 ± 3.65 0m, 5 H); 3.68 ± 3.76 0m, 3 H); 3.76 ± 3.81
0m, HÀC04)0Fuc), HÀC02)0Fuc)); 3.85 ± 3.96 0m, 4 H, including HÀC04)0Gal), HÀC03)0Fuc)); 3.97 ± 4.01
0m, 2 H); 4.49 0d, J ˆ 7.8, HÀC01)0Gal)); 4.70 0q, J ˆ 6.3, HÀC05)0Fuc)); 5.01 0d, J ˆ 3.9, HÀC01)0Fuc)); 7.58 ±
7.65 0m, 2 H); 7.74 ± 7.78 0m, 1 H); 7.93 ± 8.02 0m, 3 H); 8.28 0s, 1 H). HR-MS: 762.3334 0C38H52NO15À, [M À
Na]À; calc. 762.3337).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-2,6-dideoxy-6-{[4-0phenylmethoxy)benzoyl]amino}-d-arabino-hexitol 06m). Accord-
ing to the G.P. FC gave 21.9 mg 079%) of 6m. 1H-NMR 0400 MHz, D2O): 0.72 ± 0.90 0m, 2 H); 1.00 ± 1.20
0m, 6 H, including a d at 1.10, J ˆ 7, Me06)0Fuc)); 1.35 ± 1.62 0m, 8 H); 1.65 ± 1.72 0m, 1 H); 2.08 ± 2.18 0m, 1 H);
3.29 0dd, J ˆ 10,3, Me03)0Gal)); 3.33 ± 3.65 0m, 8 H); 3.65 ± 3.72 0m, HÀC04)0Fuc), HÀC02)0Fuc)); 3.72 ± 4.03
0m, 6 H); 4.42 0d, J ˆ 8, HÀC01)0Gal)); 4.60 0q, J ˆ 7, H ÀC05)0Fuc)); 4.90 0d, J ˆ 4, HÀC01)0Fuc)); 5.10
0s, 2 H); 7.00 0d, J ˆ 8, 2 H); 7.25 ± 7.43 0m, 5 H); 7.62 0d, J ˆ 8, 2 H). HR-MS: 818.3604 0C41H56NO16À, [M À
Na]À; calc. 818.3599).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-6-[benzoyl0phenylmethyl)amino]-2,6-dideoxy-d-arabino-hexitol 06n). According to
the G.P. FC gave 11.6 mg 048%) of 6n. 1H-NMR 0400 MHz, D2O; 2 rotamers): 0.72 ± 0.92 0m, 2 H); 0.95 ± 1.21
0m, 6 H); 1.38 ± 1.72 0m, 8 H); 1.65 ± 1.73 0m, 1 H); 1.95 ± 2.18 0m, 1 H); 3.05 ± 3.99 0m, 17 H); 4.28 ± 4.90
0m, 5 H); 7.07 ± 7.45 0m, 10 H). HR-MS: 802.3648 0C41H56NO15À, [M À Na]À; calc. 802.3650).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-2,6-dideoxy-6-[01-oxo-3-phenylpropyl)amino]-d-arabino-hexitol 06o). According to
the G.P. FC gave 17.3 mg 049%) of 6o. 1H-NMR 0400 MHz, D2O): 0.73 ± 0.92 0m, 2 H); 1.00 ± 1.21 0m, 6 H,
including a d at 1.13, J ˆ 7, Me06)0Fuc)); 1.39 ± 1.65 0m, 8 H); 1.65 ± 1.73 0m, 1 H); 2.06 ± 2.12 0m, 1 H); 2.49
0t, J ˆ 7, 2 H); 2.82 0t, J ˆ 7, 2 H); 3.18 ± 3.35 0m, 5 H); 3.50 ± 3.58 0m, 3 H); 3.62 ± 3.95 0m, 9 H, including a dd at
3.78, J ˆ 3, 11, HÀC03)0Fuc)); 4.41 0d, J ˆ 8, HÀC01)0Gal)); 4.59 0q, J ˆ 7, H ÀC05)0Fuc)); 4.78 0d, J ˆ 4,
HÀC01)0Fuc)); 7.20 0t, J ˆ 8, 3 H); 7.30 0t, J ˆ 8, 2 H). HR-MS: 740.3486 0C36H55NO15À, [M À Na]À; calc.
740.3493).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-2,6-dideoxy-6-[01-oxo-2,2-diphenylethyl)amino]-d-arabino-hexitol 06p). According to
the G.P. CC followed by FC gave 21.5 mg 079%) of 6p. 1H-NMR 0500 MHz, D2O): 0.82 ± 0.96 0m, 2 H); 1.08 ±
1.22 0m, 6 H, including a d at 1.15, J ˆ 6.5, Me06)0Fuc)); 1.46 ± 1.68 0m, 8 H); 1.72 ± 1.79 0m, 1 H); 2.11 ± 2.19
0m, 1 H); 3.45 ± 3.32 0m, 4 H); 3.62 ± 3.49 0m, 3 H); 3.65 ± 3.78 0m, 5 H); 3.82 0dd, J ˆ 3.4, 10.0, HÀC03)0Fuc));
3.87 0d, J ˆ 3.0, HÀC04)0Gal)); 3.88 ± 4.02 0m, 3 H); 4.46 0d, J ˆ 8.0, HÀC01)0Gal)); 4.64 0q, J ˆ 6.6,
HÀC05)0Fuc)); 4.78 0d, J ˆ 4.0, HÀC01)0Fuc)); 5.16 0s, Ph2CH); 7.42 ± 7.22 0m, 10 H). HR-MS: 802.3651
0C41H56NO15À, [M À Na]À; calc. 802.3650).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-6-0acetylamino)-1,5-anhydro-2,6-dideoxy-d-arabino-hexitol 06q). According to the G.P. FC gave
15.2 mg 046%) of 6q. 1H-NMR 0400 MHz, D2O): 0.73 ± 0.92 0m, 2 H); 1.00 ± 1.21 0m, 6 H, including a d at 1.13,
J ˆ 7, Me06)0Fuc)); 1.39 ± 1.65 0m, 8 H); 1.65 ± 1.73 0m, 1 H); 1.90 0s, MeCO); 2.10 ± 2.18 0m, 1 H); 3.28 0dd, J ˆ
10, 3, HÀC03)0Gal)); 3.32 ± 3.45 0m, 4 H); 3.48 ± 3.57 0m, 3 H); 3.60 ± 3.72 0m, 4 H); 3.78 0dd, J ˆ 3, 1,
HÀC03)0Fuc)); 3.81 0d, J ˆ 3, HÀC04)0Gal)); 3.83 ± 3.97 0m, 3 H); 4.40 0d, J ˆ 8, HÀC01)0Gal)); 4.62 0q, J ˆ 7,
‡
‡
HÀC05)0Fuc)); 4.80 0d, J ˆ 4, HÀC01)0Fuc)). HR-MS: 674.3002 0C29H49NNaO15 , [M ‡ H] ; calc. 674.3000).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-6-[0cyclohexylcarbonyl)amino]-2,6-dideoxy-d-arabino-hexitol 06r). According to the
G.P. FC gave 21.4 mg 089%) of 6r. 1H-NMR 0500 MHz, D2O): 0.83 ± 0.97 0m, 2 H); 1.08 ± 1.37 0m, 11 H,
including a d at 1.17, J ˆ 6.7, Me06)0Fuc)); 1.47 ± 1.68 0m, 9 H); 1.79 ± 1.68 0m, 5 H); 2.16 ± 2.27 0m, 2 H); 3.36
0dd, J ˆ 9.5, 3.2, HÀC03)0Gal)); 3.53 ± 3.40 0m, 4 H); 3.63 ± 3.55 0m, 3 H); 3.72 ± 3.68 0m, 2 H); 3.74 0dd, J ˆ
10.5, 4.0, HÀC02)0Fuc)); 3.77 0d, J ˆ 3.2, HÀC04)0Fuc)); 3.83 0dd, J ˆ 3.2, 10.5, HÀC03)0Fuc)); 3.87 0d, J ˆ 3.0,
HÀC04)0Gal)); 3.91 ± 4.05 0m, 3 H); 4.48 0d, J ˆ 8.0, HÀC01)0Gal)); 4.67 0q, J ˆ 7.0, HÀC05)0Fuc)); 4.86
0d, J ˆ 4.0, HÀC01)0Fuc)). HR-MS: 718.3650 0C34H56NO15À, [M À Na]À; calc. 718.3650).

Helvetica Chimica Acta ± Vol. 83 02000)
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O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-2,6-dideoxy-6-{[04-methylphenyl)sulfonyl]amino}-d-arabino-hexitol 06s). To a soln. of
6a 030 mg, 0.049 mmol) in sat. aq. NaHCO3 soln. 00.3 ml), 1.1m 4-methlybenzenesulfonyl chloride 0TsCl) in
toluene 050 ml, 0.054 mmol, 1.1 equiv.) was added. After 3 d, another portion of 1.1m TsCl in toluene 050 ml,
0.054 mmol, 1.1 equiv.) was added. After 6 d, the mixture was evaporated and the residue purified subsequently
by gel filtration 0P2, BioRad, H2O), FC 0silica gel, AcOEt/iPrOH/H2O 4 :2 :1), CC 0RP C18, column 1 Â 10 cm,
MeCN/H2O 3 :7, then 4 :6), and gel filtration 0P2, BioRad, H2O) and freeze-dried: 5w 016 mg, 42%). White
foam. 1H-NMR 0400 MHz, D2O): 0.75 ± 0.92 0m, 2 H); 1.00 ± 1.18 0m, 6 H, including a d at 1.08, J ˆ 7,
Me06)0Fuc)); 1.35 ± 1.60 0m, 8 H); 1.64 ± 1.72 0m, 1 H); 2.02 ± 2.08 0m, 1 H); 2.32 0s, 3 H); 3.12 0dd, J ˆ 7, 12,
1 H); 3.15 ± 3.35 0m, 4 H); 3.48 ± 3.53 0m, 2 H); 3.55 ± 3.62 0m, 3 H); 3.65 ± 3.73 0m, HÀC04)0Fuc),
HÀC02)0Fuc)); 3.72 ± 3.80 0m, HÀC03)0Fuc), HÀC04)0Gal)); 3.82 ± 3.88 0m, 2 H); 4.46 0d, J ˆ 8,
HÀC01)0Gal)); 4.53 0q, J ˆ 7, H ÀC05)0Fuc)); 4.75 0d, J ˆ 4, HÀC01)0Fuc)); 7.34 0d, J ˆ 8, 2 H); 7.04 0d, J ˆ
8, 2 H). HR-MS: 762.2998 0C34H52NO16SÀ, [M À Na]À; calc. 762.3007).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-2,6-dideoxy-6-{[0ethylamino)carbonyl]amino}-d-arabino-hexitol 06t). As described
for 6u, from 6a 030 mg, 0.049 mmol) and ethyl isocyanate instead of phenyl isocyanate. FC 0silica gel) gave
6t 021.7 mg, 64%). White foam. 1H-NMR 0400 MHz, D2O): 0.75 ± 0.92 0m, 2 H); 0.98 0t, J ˆ 7, 3 H); 1.00 ± 1.18
0m, 6 H, including a d at 1.09, J ˆ 7, Me06)0Fuc)); 1.38 ± 1.62 0m, 8 H); 1.63 ± 1.73 0m, 1 H); 2.08 ± 2.16 0m, 1 H);
3.04 0q, J ˆ 7, 2 H); 3.28 ± 3.44 0m, 5 H); 3.46 ± 3.58 0m, 3 H); 3.64 ± 3.74 0m, 4 H, including HÀC04)0Fuc),
HÀC02)0Fuc)); 3.78 0dd, J ˆ 3, 11, HÀC03)0Fuc)); 3.81 0d, J ˆ 3, HÀC04)0Gal)); 3.84 ± 3.98 0m, 3 H); 4.41
0d, J ˆ 8, HÀC01)0Gal)); 4.63 0q, J ˆ 7, H ÀC05)0Fuc)); 4.83 0d, J ˆ 4, HÀC01)0Fuc)). HR-MS: 679.3288
0C30H51N2O15À, [M À Na]À; calc. 679.3289).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-2,6-dideoxy-6-{[0phenylamino)carbonyl]amino}-d-arabino-hexitol 06u). To a soln. of
6a 017 mg, 0.027 mmol) in 0.5n NaOH 01 ml) at 08, a soln. of commercially available phenyl isocyanate 04 mg,
0.033 mmol, 1.2 equiv.) in 0.5n NaOH was added. Additional phenyl isocyanate 04 mg, 0.033 mmol, 1.2 equiv.)
was added after 1 h, and the mixture was warmed to r.t. Addition of phenyl isocyanate was continued until after
i
11 d, a total amount of 6 equiv. was added. The product was purified by FC 0silica gel; AcOEt/PrOH/H2O
‡
4 :2 :1), ion exchange 0Dowex 0Na form), H2O), gel filtration 0P2, BioRad, H2O), and again ion exchange
‡
0Dowex 0Na form), H2O) and freeze-dried: 6u 08.4 mg, 42%). White foam. 1H-NMR 0400 MHz, D2O): 0.72 ±
0.92 0m, 2 H); 0.97 ± 1.21 0m, 6 H, including a d at 1.09, J ˆ 7, Me06)0Fuc)); 1.38 ± 1.62 0m, 8 H); 1.63 ± 1.73
0m, 1 H); 2.06 ± 2.16 0m, 1 H); 3.28 ± 3.46 0m, 4 H); 3.46 ± 3.63 0m, 6 H); 3.64 ± 3.72 0m, 2 H, HÀC04)0Fuc),
HÀC02)0Fuc)); 3.76 0dd, J ˆ 3, 11, HÀC03)0Fuc)); 3.84 0s, HÀC04)0Gal)); 3.85 ± 4.08 0m, 3 H); 4.39 0d, J ˆ 8,
HÀC01)0Gal)); 4.64 0q, J ˆ 7, H ÀC05)0Fuc)); 4.86 0d, J ˆ 4, HÀC01)0Fuc)); 7.03 0t, 1 H); 7.18 0d, 2 H); 7.27
‡
0m, 2 H). HR-MS: 727.3292 0C34H51N2O15 , [M À Na]À; calc. 727.3289).
3-O-3-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-2,6-dideoxy-6-{{[04-nitrophenyl)methoxy]carbonyl}amino}-d-arabino-hexitol 06v).
According to the G.P. from 6a 030 mg). FC followed by gel filtration gave 6v 020 mg, 50%). 1H-NMR
0400 MHz, D2O): 0.73 ± 0.90 0m, 2 H); 1.03 ± 1.21 0m, 6 H, including a d at 1.13, J ˆ 7, Me06)0Fuc)); 1.39 ± 1.60
0m, 8 H); 1.65 ± 1.72 0m, 1 H); 2.08 ± 2.15 0m, 1 H); 3.28 ± 3.42 0m, 5 H); 3.48 ± 3.58 0m, 3 H); 3.59 ± 3.78
0m, 5 H); 3.81 0dd, J ˆ 3, 10, HÀC03)0Fuc)); 3.84 ± 3.98 0m, 3 H); 4.40 0d, J ˆ 8, HÀC01)0Gal)); 4.58 0q, J ˆ 7,
HÀC05)0Fuc)); 4.80 0d, J ˆ 4, HÀC01)0Fuc)); 5.12 0m, PhCH2); 7.48 0d, J ˆ 8, 2 H); 8.16 0d, J ˆ 7, 2 H). HR-
MS: 787.3136 0C35H51N2O18À, [M À Na]À; calc. 787.3137).
3-O-3-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-2,6-dideoxy-6-{[0naphthalen-2-ylmethoxy)carbonyl]amino}-d-arabino-hexitol 06w).
According to the G.P. FC followed by gel filtration gave 12 mg 030%) of 6w. 1H-NMR 0400 MHz, D2O):
0.74 ± 0.94 0m, 2 H); 0.95 ± 1.18 0m, 6 H, including a d at 1.08, J ˆ 7, Me06)0Fuc)); 1.38 ± 1.60 0m, 8 H); 1.64 ± 1.72
0m, 1 H); 2.05 ± 2.14 0m, 1 H); 3.28 ± 3.42 0m, 5 H); 3.48 ± 3.58 0m, 3 H); 3.59 ± 3.78 0m, 5 H); 3.78 ± 3.98
0m, 4 H); 4.38 0d, J ˆ 8.0, HÀC01)0Gal)); 4.59 0q, J ˆ 6.3, HÀC05)0Fuc)); 4.81 0d, J ˆ 3.8, HÀC01)0Fuc)); 5.18
‡
‡
0m, PhCH2); 7.38 ± 7.52 0m, 3 H); 7.78 ± 7.88 0m, 4 H). HR-MS: 816.3422 0C39H55NNaO16 , [M ‡ H] ; calc.
816.3419).
3-O-3-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-a-l-galactopyrano-
syl-01 ! 4)]-1,5-anhydro-2,6-dideoxy-6-{[0phenylmethoxy)carbonyl]amino}-d-arabino-hexitol 06x). According
to the G.P. CC followed by FC gave 18 mg 072%) of 6x. 1H-NMR 0500 MHz, D2O): 0.80 ± 0.94 0m, 2 H); 1.03 ±
1.21 0m, 6 H, including a d at 1.13, J ˆ 6.4, Me06)0Fuc)); 1.44 ± 1.65 0m, 8 H); 1.71 ± 1.76 0m, 1 H); 2.12 ± 2.18
0m, 1 H); 3.33 0dd, J ˆ 9.6, 3.6, HÀC03)0Gal)); 3.35 ± 3.44 0m, 4 H); 3.54 ± 3.62 0m, 3 H); 3.66 ± 3.76 0m, 4 H);

2904
Helvetica Chimica Acta ± Vol. 83 02000)
3.81 0dd, J ˆ 3.0, 10.4, HÀC03)0Fuc)); 3.85 0d, J ˆ 3.0, HÀC04)0Gal)); 3.89 ± 3.95 0m, 2 H); 3.95 ± 4.01 0m, 1 H);
4.45 0d, J ˆ 8.0, HÀC01)0Gal)); 4.66 0q, J ˆ 6.3, HÀC05)0Fuc)); 4.86 0d, J ˆ 3.8, HÀC01)0Fuc)); 5.06
0m, PhCH2); 7.42 ± 7.33 0m, 5 H). HR-MS: 742.3282 0C35H52NO16À, [M À Na]À; calc. 742.3286).
O-3-O-[01S)-1-Cyclohexylmethyl)-2-oxo-2-0phenylmethoxy)ethyl]-2,4,6-tris-O-0phenylmethyl)-b-d-galac-
topyranosyl-01 ! 3)-O-[6-deoxy-2,3,4-tris-O-0phenylmethyl)-a-l-galactopyranosyl-01 ! 4)]-1,5-anhydro-2-de-
oxy-d-arabino-hexitol 07). A soln. of Br2 01.90 g, 11.8 mmol) in CH2Cl2 011 ml) was added dropwise at 08 to a
soln. of 11 04.5 g, 9.45 mmol) in CH2Cl2 011 ml). After stirring for 30 min at 08, cyclohexene 02.5 ml) was added
to consume excessive Br2. The soln. was added within 10 min to a mixture of 16 07.71 g, 7.87 mmol), Et4NBr
02.00 g, 9.45 mmol; dried for 2 h at 2008), and molecular sieves 012 g; dried for 24 h at 3008) in DMF/CH2Cl2 1:1
060 ml). The mixture was stirred for 65 h at 208, diluted with CH2Cl2 0250 ml), and filtered. The resulting soln.
was washed with NaHCO3 soln. 02 Â 50 ml), H2O 02 Â 250 ml), 00.5m HCl, 2 Â 250 ml), and brine 0250 ml),
dried 0Na2SO4) and evaporated. To the residue dissolved in MeCN 085 ml) at 208, a soln. of Et3N 00.21 ml) and
H2SiF6 01.3 ml, 35%) in MeCN 017 ml) was added within 10 min. After stirring for 2 h, the mixture was diluted
with CH2Cl2 0250 ml), washed with NaHCO3 soln. 03 Â 250 ml) and brine 0250 ml), dried 0Na2SO4), and
evaporated, and the residue subjected to CC 0silica gel, hexane/AcOEt 1:1 ! 1 :2): 7 07.88 g, 78%). Colorless
solid. 1H-NMR 0400 MHz, CDCl3): 0.45 ± 1.44 0m, 14 H, HaxÀC02)0Hex), CH20Chx)); 1.39 0d, J ˆ 6.5,
Me06)0Fuc)); 1.89 0m, HeqÀC02)0Chx)); 2.25 0t, J ˆ 6.5, OHÀC06)0Hex)); 3.03 0dt, J ˆ 9.0, 3.5,
HÀC05)0Hex)); 3.20 0br. t, J ˆ 12.0, HaxÀC01)0Hex)); 3.52 0br. d, J ˆ 3.0, HÀC04)0Fuc)); 3.55 0t, J ˆ 9.0,
HÀC04)0Hex)); 3.74 0m, HeqÀC01)0Hex), HÀC03)0Hex), 2 HÀC06)0Hex)); 3.87 0dd, J ˆ 10.0, 3.5,
HÀC03)0Gal)); 3.94 0br. t, J ˆ 6.5, HÀC05)0Gal)); 3.99 0dd, J ˆ 10.0, 2.5, Me03)0Fuc)); 4.07 0dd, J ˆ 10.0,
3.5, HÀC02)0Fuc)); 4.15 0dd, J ˆ 8.0, 4.5, OCHCO2Bn); 4.32 0dd, J ˆ 11.5, 7.5, HaÀC06)0Gal)); 4.34 0d, J ˆ 11.5,
1 H, PhCH2); 4.42 0dd, J ˆ 11.5, 5.5, HbÀC06)0Gal)); 4.55 0d, J ˆ 12.0, 1 H, PhCH2); 4.61 0d, J ˆ 8.0,
HÀC01)0Gal)); 4.63 0d, J ˆ 12.0, 1 H, PhCH2); 4.69 0br. q, J ˆ 6.5, HÀC05)0Fuc)); 4.69 0d, J ˆ 11.5, 1 H,
PhCH2); 4.79 0d, J ˆ 11.5, 1 H, PhCH2); 4.83 0d, J ˆ 11.5, 1 H, PhCH2); 5.04 0d, J ˆ 3.5, HÀC01)0Fuc)); 5.07
0d, J ˆ 12.0, 1 H, PhCH2); 5.14 0d, J ˆ 12.0, 1 H, PhCH2); 5.62 0dd, J ˆ 10.0, 8.0, HÀC02)0Gal)); 5.87 0dd, J ˆ 3.5,
‡
‡
0.5, HÀC04)0Gal)); 7.17 ± 8.14 0m, 35 arom. H). HR-MS: 1305.5389 0C76H82NaO18
;
[M ‡ Na] ; calc.
1305.5399).
Ethyl 3-O-[01S)-1-0Cyclohexylmethyl)-2-oxo-2-0phenylmethoxy)ethyl]-1-thio-b-d-galactopyranoside 012).
A suspension of 8 015.0 g, 66.9 mmol) and Bu2SnO 020.0 g, 80.3 mmol) in MeOH 0450 ml) was refluxed for 2 h
0 ! clear soln.). Evaporation and the repeated co-evaporation with pentane gave a colorless foam, which was
dried under vacuum for 1 h. MeOCH2CH2OMe 0120 ml), 9 039.6 g, 100.3 mmol) dissolved in MeOCH2-
CH2OMe 060 ml), and CsF 012.2 g, 80.3 mmol; dried) were added, and the suspension was stirred for 2 h at 208.
Then 1m KH2PO4 0700 ml) and KF 025 g) were added, followed by extraction with AcOEt 03 Â 250 ml). The
combined org. extract was washed with 10% KF soln. 02 Â 250 ml) and brine 01 Â 250 ml), dried 0Na2SO4), and
evaporated, and the residue subjected to FC 0silica gel, hexane/acetone 4 :1 ! 2 :1): 12 020.4 g, 65%). Colorless
solid. 1H-NMR 0400 MHz, CD3OD): 0.82 ± 1.88 0m, 13 H, CH20Chx)); 1.28 0t, J ˆ 7.5, MeCH2S); 2.74
0m, MeCH2S); 3.25 0dd, J ˆ 9.0, 3.0, HÀC03)0Gal)); 3.45 0ddd, J ˆ 7.0, 5.0, 1.0, HÀC05)0Gal)); 3.61 0dd, J ˆ
11.5, 5.0, HaÀC06)0Gal)); 3.66 0t, J ˆ 9.5, HÀC02)0Gal)); 3.71 0dd, J ˆ 11.5, 7.0, HbÀC06)0Gal)); 3.92 0dd, J ˆ
3.0, 1.0, HÀC04)0Gal)); 4.28 0d, J ˆ 10.0, HÀC01)0Gal)); 4.48 0dd, J ˆ 8.5, 4.0, OCHCO2Bn); 5.14 0d, J ˆ 12.0,
‡
1 H, PhCH2); 5.25 0d, J ˆ 12.0, 1 H, PhCH2); 7.34 ± 7.40 0m, 5 arom. H). ES-MS: 486 0[M ‡ NH4] ).
Ethyl 3-O-[01S)-1-0Cyclohexylmethyl)-2-oxo-2-0phenylmethoxy)ethyl]-1-thio-b-d-galactopyranoside 2,4,6-
Tribenzoate 013). At 08, benzoyl chloride 052.1 g, 370.7 mmol) was added to a soln. of 12 019.3 g, 41.2 mmol) and
DMAP 01.51 g) in pyridine 0210 ml). The mixture was stirred at 208 for 6 h, the solvent removed, and the residue
dissolved in AcOEt 0500 ml). The soln. was extracted with 0.1m HCl 02 Â 250 ml), NaHCO3 soln. 02 Â 250 ml),
and brine 0250 ml), dried 0Na2SO4), and evaporated, and the residue subjected to FC 0silica gel, hexane/AcOEt
6 :1 ! 4 :1): 13 028.9 g, 90%). Colorless solid. 1H-NMR 0400 MHz, CDCl3): 0.44 ± 1.43 0m, 13 H, CH20Chx));
1.26 0t, J ˆ 7.5, MeCH2S); 2.77 0m, MeCH2S); 3.93 0dd, J ˆ 9.5, 3.5, HÀC03)0Gal)); 4.02 0ddd, J ˆ 7.0, 5.0, 1.0,
HÀC05)0Gal)); 4.21 0dd, J ˆ 8.0, 5.0, OCHCO2Bn); 4.42 0dd, J ˆ 11.5, 5.0, HaÀC06)0Gal)); 4.49 0dd, J ˆ 11.5,
7.0, HbÀC06)0Gal)); 4.62 0d, J ˆ 10.0, HÀC01)0Gal)); 5.08 0d, J ˆ 12.0, 1 H, PhCH2); 5.21 0d, J ˆ 12.0, 1 H,
PhCH2); 5.69 0t, J ˆ 9.5, HÀC02)0Gal)); 6.00 0dd, J ˆ 3.5, 1.0, HÀC04)0Gal)); 7.30 ± 8.17 0m, 20 arom. H). HR-
‡
‡
MS: 803.2857 0C45H48NaO10S , [M ‡ Na] ; calc. 803.2866).
1,5-Anhydro-2-deoxy-3-O-[2,4,6-tri-O-benzoyl-3-O-[01S)-1-0cyclohexylmethyl)-2-oxo-2-0phenylmethoxy)-
ethyl]-b-d-galactopyranosyl]-d-arabino-hexitol 014). At À108, 0.15m CF3SO3H soln. in CH2Cl2 was added to a
soln. of 10 06.05 g, 25.64 mmol) and 13 010.0 g, 12.82 mmol) in CH2Cl2 075 ml). The addition was stopped when
the orange soln. turned brown. AcOEt was added 0500 ml), the mixture extracted with NaHCO3 soln. 04 Â
250 ml) and brine 0250 ml), dried 0Na2SO4), and evaporated, and the residue subjected to FC 0silica gel, Et2O):

Helvetica Chimica Acta ± Vol. 83 02000)
2905
14 09.03 g, 81%). Colorless solid. 1H-NMR 0400 MHz, CDCl3): 0.47 ± 1.46 0m, 13 H, CH20Chx)); 1.52 ± 1.68
0m, 2 HÀC02)0Hex)); 2.12 0br. t, J ˆ 6.0, OHÀC06)0Hex)); 3.13 0ddd, J ˆ 9.0, 5.5, 4.0, HÀC05)0Hex)); 3.30
0td, J ˆ 12.0, 2.0, HaxÀC01)0Hex)); 3.42 0t, J ˆ 9.0, HÀC04)0Hex)); 3.47 0m, HÀC03)0Hex)); 3.68 0dt, J ˆ 11.0,
5.5, HaÀC06)0Hex)); 3.83 0m, HeqÀC01)0Hex), HbÀC06)0Hex)); 3.91 0dd, J ˆ 10.0, 3.5, HÀC03)0Gal)); 4.03
0br. dd, J ˆ 9.0, 3.0, HÀC05)0Gal)); 4.21 0dd, J ˆ 8.0, 5.0, OCHCO2Bn); 4.25 0br. s, OHÀC04)0Hex)); 4.28
0dd, J ˆ 12.0, 9.0, HaÀC06)0Gal)); 4.64 0d, J ˆ 8.0, HÀC01)0Gal)); 4.68 0dd, J ˆ 12.0, 3.0, HbÀC06)0Gal)); 5.10
0d, J ˆ 12.0, 1 H, PhCH2); 5.23 0d, J ˆ 12.0, 1 H, PhCH2); 5.67 0dd, J ˆ 10.0, 8.0, HÀC02)0Gal)); 5.91 0br. d, J ˆ
‡
‡
3.5, HÀC04)0Gal)); 7.33 ± 8.17 0m, 20 arom. H). HR-MS: 889.3409 0C49H54NaO14 , [M ‡ Na] ; calc. 889.3411).
In addition to 14, 10% of the a-d-epimer 15 was isolated. 1H-NMR 0400 MHz, CDCl3): 0.48 ± 1.68 0m,
14 H, HaxÀC02)0Hex), CH20Chx)); 1.90 0m, OHÀC06)0Hex)); 2.03 0m, HeqÀC02)0Hex)); 2.23 0d, J ˆ 1.0,
OHÀC04)0Hex)); 3.12 0m, HÀC05)0Hex)); 3.23 0td, J ˆ 12.0, 2.0, HaxÀC01)0Hex)); 3.45 ± 3.58 0m,
HeqÀC01)0Hex), HÀC03)0Hex), HÀC04)0Hex)); 3.67 0br. d, J ˆ 11.0, HaÀC06)0Hex)); 3.78 0br. d, J ˆ 11.0,
HbÀC06)0Hex)); 4.15 0dd, J ˆ 9.5, 3.5, HÀC03)0Gal)); 4.36 0dd, J ˆ 8.0, 5.0, OCHCO2Bn); 4.39 ± 4.45
0m, HÀC05)0Gal), HaÀC06)0Gal)); 4.48 0dd, J ˆ 10.0, 2.0, HbÀC06)0Gal)); 5.18 0d, J ˆ 12.0, 1 H, PhCH2);
5.32 0d, J ˆ 12.0, 1 H, PhCH2); 5.55 0m, HÀC01)0Gal), HÀC02)0Gal); 5.99 0br. d, J ˆ 3.5, HÀC04)0Gal));
‡
7.34 ± 8.15 0m, 20 arom. H). ESI-MS: 884 0[M ‡ NH4] ).
1,5-Anhydro-2-deoxy-6-O-[01,1-dimethylethyl)dimethylsilyl]-3-O-{2,4,6-tri-O-benzoyl-3-O-[01S)-1-0cyclo-
hexylmethyl)-2-oxo-2-0phenylmethoxy)ethyl]-b-d-galactopyranosyl]-d-arabino-hexitol 016). A soln. of 14
t
07.96 g, 9.19 mmol), BuMe2SiCl 01.52 g, 10.1 mmol) and 1H-imidazol 00.94 g, 13.8 mmol) in DMF 055 ml)
was stirred for 1 h at 208. The mixture was diluted with AcOEt 0250 ml), washed with NaHCO3 soln. 05 Â
250 ml) and brine 0250 ml), dried 0Na2SO4), and evaporated, and the residue subjected to FC 0silica gel, hexane/
AcOEt 4 :1 ! 1:1): 16 08.38 g, 93%). Colorless solid. 1H-NMR 0400 MHz, CDCl3): 0.01, 0.03 02s, Me2Si); 0.45 ±
1.64 0m, 15 H, 2 HÀC02)0Hex), CH20Chx)); 0.87 0s, tBuSi); 3.05 0ddd, J ˆ 9.0, 5.5, 1.5, HÀC05)0Hex)); 3.23
0td, J ˆ 12.0, 2.0, HaxÀC01)0Hex)); 3.39 0t, J ˆ 9.0, HÀC04)0Hex)); 3.45 0m, HÀC03)0Hex)); 3.73 0dt, J ˆ 11.5,
5.5, HaÀC06)0Hex)); 3.82 0br. dd, J ˆ 12.0, 3.5, HeqÀC01)0Hex)); 3.89 0dd, J ˆ 10.0, 3.5, HÀC03)0Gal)); 3.90
0dd, J ˆ 11.5, 1.5, HbÀC06)0Hex)); 3.96 0br. s, OHÀC04)0Hex)); 4.02 0br. dd, J ˆ 8.5, 3.5, HÀC05)0Gal)); 4.19
0dd, J ˆ 8.0, 4.5, OCHCO2Bn); 4.31 0dd, J ˆ 12.0, 8.5, HaÀC06)0Gal)); 4.62 0dd, J ˆ 12.0, 3.5, HbÀC06)0Gal));
4.64 0d, J ˆ 8.0, HÀC01)0Gal)); 5.08 0d, J ˆ 12.0, 1 H, PhCH2); 5.21 0d, J ˆ 12.0, 1 H, PhCH2); 5.66 0dd, J ˆ 10.0,
8.0, HÀC02)0Gal)); 5.91 0d, J ˆ 3.5, HÀC04)0Gal)); 7.29 ± 8.17 0m, 20 arom. H). HR-MS: 1003.4289
‡
‡
0C55H68NaO14Si , [M ‡ Na] ; calc. 1003.4276).
O-3-O-{2,4,6-Tri-O-benzoyl-3-O-[01S)-1-0cyclohexylmethyl)-2-oxo-2-0phenylmethoxy)ethyl]-b-d-galacto-
pyranosyl-01 ! 3)-O-[6-deoxy-2,3,4-tris-O-0phenylmethyl)-a-l-galactopyranosyl-01 ! 4)]-1,5-anhydro-2-de-
oxy-6-O-0methylsulfonyl)-d-arabino-hexitol 017). To a soln. of 7 012.5 g, 9.75 mmol) in dry pyridine 080 ml),
methanesulfonyl chloride 03.35 g, 29.2 mmol) was added dropwise with magnetic stirring within 5 min 0Ar, r.t.).
After 30 min, the mixture was diluted with AcOEt 0500 ml) and extracted with 1n HCl 0250 ml). The aq. phase
was extracted twice with AcOEt 0300 ml), the combined org. phase 0Na2SO4) dried and evaporated, and the
residue submitted to FC 0silica gel 0500 g), hexanes/AcOEt 6 :4): 12.98 g 097%) of 17. 1H-NMR 0400 MHz,
CDCl3): 0.4 ± 1.4 0m, 14 H, HaxÀC02)0Hex), CH20Chx)); 1.39 0d, J ˆ 6.5, HÀC06)0Fuc)); 1.85 ± 1.95 0m, Heq
À
C02)0Hex)); 2.89 0s, MeSO3ÀC06)); 3.08 0m, HÀC05)0Hex)); 3.15 0t, J ˆ 11, HaxÀC01)0Hex)); 3.45 ± 3.52
0m, HÀC04)0Hex), HÀC04)0Fuc)); 3.6 ± 3.7 0m, HÀC03)0Hex)); 3.75 0br. d, J ˆ 11, HeqÀC01)0Hex)); 3.85
0dd, J ˆ 10.0, 3.5, HÀC03)0Gal)); 3.9 ± 4.0 0m, HÀC05)0Gal), HÀC03)0Fuc)); 4.03 0dd, J ˆ 10.0, 3.5,
HÀC02)0Fuc)); 4.12 0dd, J ˆ 8.0, 4.5, OCHCO2Bn); 4.27 ± 4.37 0m, HaÀC06)0Hex), 1 H of PhCH2); 4.37 ±
4.45 0m, HbÀC06)0Hex), HaÀC06)0Gal)); 4.48 0dd, J ˆ 11.5, 5.5, HbÀC06)0Gal)); 4.52 ± 4.68 05 H,
HÀC01)0Gal), HÀC05)0Fuc), PhCH2); 4.78, 4.83 02d, 2 H, PhCH2); 4.93 0d, J ˆ 3.5, HÀC01)0Fuc)); 5.05,
5.13 02d, J ˆ 12, CO2CH2Ph); 5.60 0dd, J ˆ 10.0, 8.0, HÀC02)0Gal)); 5.83 0d, J ˆ 3.5, HÀC04)0Gal)); 7.13 ± 7.35
0m, 22 H); 7.35 ± 7.42 0m, 5 H); 7.45 ± 7.5 0m, 2 H); 8.0 ± 8.13 0m, 6 arom. H). HR-MS: 1383.5168
‡
‡
0C77H84NaO20S , [M ‡ Na] ; calc. 1383.5174).
O-3-O-{2,4,6-Tri-O-benzoyl-3-O-[01S)-1-0cyclohexylmethyl)-2-oxo-2-0phenylmethoxy)ethyl]-b-d-galacto-
pyranosyl-01 ! 3)-O-[6-deoxy-2,3,4-tris-O-0phenylmethyl)-a-l-galactopyranosyl-01 ! 4)]-1,5-anhydro-6-azido-
2,6-dideoxy-d-arabino-hexitol 018). To a soln. of 17 012.95 g, 9.52 mmol) in DMF 040 ml), NaN3 04.64 g,
74.4 mmol) was added. The mixture was heated to 658 under Ar and stirred for 35 h. After cooling, the soln. was
diluted with AcOEt 0500 ml) and washed with H2O 0300 ml) and saturated brine 0150 ml). The aq. phases
were extracted with AcOEt 02 Â 300 ml). The combined org. phase was dried 0Na2SO4) and evaporated
and the residue submitted to FC 0silica gel 0500 g), hexanes/AcOEt 1:1): 12.2 g 098%) of 18. 1H-NMR
0400 MHz, CDCl3): 0.4 ± 1.5 0m, 14 H, HaxÀC02)0Hex), Ch20Chx)); 1.40 0d, J ˆ 6.5, HÀC06)0Fuc)); 1.85 ± 1.95
0m, HeqÀC02)0Hex)); 3.05 ± 3.13 0m, HÀC05)0Hex)); 3.17 0br. t, J ˆ 11, HaxÀC01)0Hex)); 3.42 0dd, J ˆ 12, 5,

2906
Helvetica Chimica Acta ± Vol. 83 02000)
HaÀC06)0Hex)); 3.45 ± 3.55 0m, HÀC04)0Hex), HbÀC06)0Hex), HÀC04)0Fuc)); 3.6 ± 3.7 0m, HÀC03)0Hex));
3.78 0br. d, J ˆ 11, HeqÀC01)0Hex)); 3.86 0dd, J ˆ 10.0, 3.5, HÀC03)0Gal)); 3.9 ± 4.0 0m, HÀC05)0Gal),
HÀC03)0Fuc)); 4.03 0dd, J ˆ 10.0, 2.5, HÀC02)0Fuc)); 4.13 0dd, J ˆ 8.0, 4.5, OCHCO2Bn); 4.33 0dd, J ˆ 11.5,
7.5, HaÀC06)0Gal)); 4.33 0d, J ˆ 11.5, 1 H); 4.77 0d, J ˆ 11.5, 1 H, PhCH2); 4.40 0dd, J ˆ 11.5, 5.5,
HbÀC06)0Gal)); 4.54, 4.62 02 d, each J ˆ 11.5, PhCH2); 4.58 0d, J ˆ 8, HÀC01)0Gal)); 4.63, 4.83 02d, each J ˆ
11.5, PhCH2); 4.65 ± 4.73 0m, HÀC05)0Fuc)); 4.87 0d, J ˆ 3.5, HÀC01)0Fuc)); 5.07, 5.15 02d, each J ˆ 11.5,
CO2CH2Ph); 5.63 0dd, J ˆ 10.0, 8.0, HÀC02)0Gal)); 5.86 0d, J ˆ 3.5, HÀC04)0Gal)); 7.17 ± 7.4 0m, 22 arom. H);
7.4 ± 7.5 0 m, 5 arom. H); 7.55 ± 7.65 0m, 2 arom. H); 8.0 ± 8.15 0m, 6 arom. H). HR-MS: 1330.5460
‡
‡
0C76H81NaN3O17 , [M ‡ Na] ; calc. 1330.5464).
O-3-O-[01S)-1-Carboxy-2-cyclohexylethyl]-b-d-galactopyranosyl-01 ! 3)-O-[6-deoxy-2,3,4-tri-O-0phenyl-
methyl)-a-l-galactopyranosyl-01 ! 4)]-1,5-anhydro-6-azido-2,6-dideoxy-d-arabino-hexitol 019). A soln./sus-
pension of 18 012.2 g, 9.33 mmol) and LiOH ´ H2O 05.1 g, 121.3 mmol) in MeOH 0200 ml) and H2O 020 ml)
was stirred and heated to 658 under Ar under reflux. After 20 h, the mixture was concentrated under vacuum to
1/3 of the volume. After addition of Et2O 0500 ml) and sat. brine 0200 ml), the aq. phase was extracted with Et2O
02 Â 300 ml). The org. phases were washed with brine 0150 ml), dried 0Na2SO4), and evaporated. FC 0silica gel
0350 g) hexanes/AcOEt 7:3, then CHCl3/MeOH/AcOH 94 :5 :1) of the residue 012.2 g) gave 8.10 g 096%) of 19.
1H-NMR 0400 MHz, CDCl3): 0.8 ± 1.05 0m, 2 H), 1.05 ± 1.35 0m, 3 H), and 1.55 ± 1.9 0m, 9 H, HaxÀC02)0Hex),
CH20Chx)); 1.17 0d, J ˆ 6.5, Me06)0Fuc)); 2.03 ± 2.16 0m, HeqÀC02)0Hex)); 3.23 ± 3.4 0m, 3 H), 3.4 ± 3.6
0m, 4 H), and 3.7 ± 4.05 0m, 11 H, HÀC01)0Hex), HÀC03)0Hex), HÀC04)0Hex), HÀC05)0Hex,
2 HÀC06)0Hex), HÀC02)0Gal), HÀC03)0Gal), HÀC04)0Gal), HÀC05)0Gal), OHÀC02)0Gal), OHÀC04)
0Gal), OHÀC06)0Gal), HÀC03)0Fuc), HÀC04)0Fuc)); 4.26 ± 4.44 0m, HÀC01)0Gal), OCHCO2H); 4.45
0q, J ˆ 6.5, HÀC05)0Fuc)); 4.55 ± 5.0 0m, 6 H, PhCH2); 4.95 0d, J ˆ 3.5, HÀC01)0Fuc)); 7.2 ± 7.4 0m, 15 ar-
‡
‡
om. H). HR-MS: 928.4203 0C48H63NaN3O14 , [M ‡ Na] ; calc. 928.4208).
We wish to thank Mrs. Christine Bourquin, Mrs. Brigitte Furrer, Mr. Kurt Martin, and Mr. Franz
Schwarzenbach for excellent technical support.
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Received April 10, 2000