1H-1,2,3-Triazol-1-yl thiodigalactoside derivatives as high affinity galectin-3 inhibitors

Article abstract of DOI:10.1016/j.bmc.2010.05.040

Galactose C3-triazole derivatives were synthesized by Cu(I)-catalyzed cycloaddition between acetylenes and galactose C3-azido derivatives. Evaluation against galectin-3, 7, 8N (N-terminal) and 9N (N-terminal) revealed 1,4-disubstituted triazoles to be high-affinity inhibitors of galectin-3 with selectivity over galectin-7, 8N, and 9N. Conformational analysis of 1,4-di- and 1,4,5-tri-substituted galactose C3-triazoles suggested that a triazole C5-substituent interfered sterically with the galectin proteins, which explained their poor affinities compared to the corresponding 1,4-disubstituted triazoles. Introduction of two 1,4-disubstituted triazole moieties onto thiodigalactoside resulted in affinities down to 29 nM for galectin-3.

Full text of DOI:10.1016/j.bmc.2010.05.040

Bioorganic & Medicinal Chemistry 18 (2010) 5367–5378
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
1H-1,2,3-Triazol-1-yl thiodigalactoside derivatives as high affinity galectin-3
inhibitors
Bader A. Salameh a, , Ian Cumpstey b, Anders Sundin b, Hakon Leffler c,à, Ulf J. Nilsson b,
*
a Chemistry Department, The Hashemite University, PO Box 150459, Zarka 13115, Jordan
b Organic Chemistry, Lund University, PO Box 124, SE-221 00 Lund, Sweden
c Section MIG, Department of Laboratory Medicine, University of Lund, Sölvegatan 23, SE-223 62 Lund, Sweden
a r t i c l e i n f o
a b s t r a c t
Article history:
Galactose C3-triazole derivatives were synthesized by Cu(I)-catalyzed cycloaddition between acetylenes
and galactose C3-azido derivatives. Evaluation against galectin-3, 7, 8N (N-terminal) and 9N (N-terminal)
revealed 1,4-disubstituted triazoles to be high-affinity inhibitors of galectin-3 with selectivity over galec-
tin-7, 8N, and 9N. Conformational analysis of 1,4-di- and 1,4,5-tri-substituted galactose C3-triazoles sug-
gested that a triazole C5-substituent interfered sterically with the galectin proteins, which explained
their poor affinities compared to the corresponding 1,4-disubstituted triazoles. Introduction of two
1,4-disubstituted triazole moieties onto thiodigalactoside resulted in affinities down to 29 nM for galec-
tin-3.
Received 5 October 2009
Revised 13 May 2010
Accepted 14 May 2010
Available online 20 May 2010
Keywords:
Galectin
Azide
Triazole
Inhibition
Galactose
Ó 2010 Elsevier Ltd. All rights reserved.
1. Introduction
binding glycoconjugates decrease metastasis in mice through inhi-
bition of galectins.12,13 More recently, potent efficacy of synthetic
The galectins are a family of carbohydrate binding proteins
known by their affinity for b-galactosides.1,2 Fourteen members
of galectin family have been discovered so far. They are widely dis-
tributed in vertebrates (fish, birds, amphibians, and mammals),
and in invertebrates (worms and insects). They have also been
found in protists (sponges and fungi). All galectins share a core se-
quence consisting of about 130 amino acids, many of which are
highly conserved. Galectins are soluble proteins and possess prop-
erties expected for both extra- and intracellular proteins. Galectin
properties, of which a majority involves glycoconjugate binding,
have been studied extensively due to the growing evidence of links
to immunity, inflammation,3 and cancer.4
In recent years several studies have initiated the emergence of a
coherent picture concerning the galectins’ biological mechanisms.
A central mechanism is the regulation of cell-surface receptors
via lattice formations5–8 and to intracellular glycoprotein traffick-
ing.9,10 The importance of galectins in regulating activities of cell
surface receptors, such as TGF-b or TCR, may be linked to effects
of galectin inhibition in animal models. A fragment of galectin-3
containing the CRD inhibited breast cancer in a mouse model by
acting as a dominant negative galectin-3 inhibitor11 and galectin-
galectin inhibitors clinically relevant in vitro and ex vivo cell assays
has been demonstrated, including attenuation of tumor cell motil-
ity,14 inhibition of alternative macrophage differentiation and
fibrosis,15 and enhancement of sensitivity to chemo- and radio-
therapy by activation of caspase-3-mediated apoptosis.16 Such
observations clearly suggest that efficient and selective inhibitors
for galectins can act intracellularly and find potential use in mod-
ulating inflammatory processes and cancer growth.17
We have reported the rational design of potent inhibitors of
galectin-3, by exploiting the X-ray crystal structure of N-acetyllac-
tosamine-galectin complex.18 This complex shows an extending
binding groove close to the LacNAc HO30 that could be targeted
with aromatic amides attached to C30 of LacNAc that interact with
Arg144.19,20 Derivatization at C3 of galactose is also expected to re-
sult in the added advantage of conferring selectivity over other gal-
actose-recognizing proteins, as these typically recognize terminal
galactose residues with free HO3 and HO4. In contrast, galectins
can recognize internal galactose with C3 glycosylated and hence
tolerate synthetic C3-substituents. Aromatic diamides of thiodiga-
lactoside were subsequently discovered to possess even further en-
hanced affinity for several members of the galectin family of
proteins, which was hypothesized to be a result of both aromatic
amides stacking with arginine side-chains.21,22 The aromatic
amides at LacNAc C30 or at thiodigalactoside C3 and C30 were all
prepared from a common galactose C3-azido derivative. However,
the azido functionality is versatile in the sense that it, in addition
* Corresponding author. Tel.: +46 46 222 8218; fax: +46 46 222 8209.
E-mail addresses: bader04@gmail.com (B.A. Salameh), hakon.leffler@med.lu.se
(H. Leffler), ulf.nilsson@organic.lu.se (U.J. Nilsson).
Tel.: +962 5 3903333x4126; fax: +962 5 3826613.
Tel.: +46 46 173 270; fax: +46 46 137468.
à
0968-0896/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2010.05.040

5368
B. A. Salameh et al. / Bioorg. Med. Chem. 18 (2010) 5367–5378
to for example, reduction–acylation, can undergo various mild and
selective organic transformations. In particular, the Cu(I)-catalyzed
regioselective dipolar cycloaddition between azides and acetylenes
to give 1,4-triazoles23–25 has found widespread use and has be-
come important in drug discovery.26,27 The stability of the triazole
toward oxidation, reduction and hydrolysis,28 the efficient Cu(I)-
catalyzed reaction, as well as the 1,2,3-triazole ring being an bio-
isostere to the amide-functionality in our earlier inhibitors, at-
tracted our attention for development of novel galectin
inhibitors. We have reported preliminary accounts on our efforts
on exploiting the azido group of the galactose C3-azido derivative
1 in cycloadditions with monosubstituted acetylene derivatives to-
wards galectin-3 inhibitors29 and that the acetylene galactose C3-
azido cycloaddition can be used in combination with other chem-
istries to discover galectin inhibitors via a fragment-based strat-
egy.30 We herein wish to report our continued study on 3-
triazol-1-yl-galactosides29 as galectin inhibitors including synthe-
sis of 1,5-disubstituted and 1,4,5-trisubstituted triazoles, as well
as 30-triazol-1-yl N-acetyllactosamine (LacNAc) and 3,30-di-tria-
zol-1-yl-thiodigalactoside derivatives. Furthermore, the galectins,
as many medically relevant protein targets, exist as a class of target
proteins with structural similarities, which makes it imperative to
address the, in terms of in vivo inhibitor use, important issue of
galectin selectivity. Hence, we herein extended our evaluations of
the triazol-1-yl galactoside and LacNAc derivatives against galec-
tin-7, 8N (N-terminal domain), and 9N (N-terminal domain). Final-
ly, conformational analysis of the triazol-1-yl galactoside
derivatives provided insight into structure–activity relationships
and galectin-selectivities for 1,4-di- and 1,4,5-tri-substituted tria-
zoles, respectively.
2. Results and discussion
2.1. Synthesis
A collection of triazoles was obtained on the 3-Gal position
either by the thermal 1,3-dipolar cycloaddition or by Cu(I)-cata-
lyzed reaction depending on the substituents (Scheme 1). The
monosubstituted triazole 2 was obtained by the heating of the
azide 119 with propiolic acid under which conditions the initially
formed 4-carboxytriazole intermediate decarboxylated in situ.31
The disubstituted triazoles 3–12 were obtained through the Cu(I)
catalyzed reaction of the azide 1 with monosubstituted acetylenes
as earlier reported.29 The reactions were highly regioselective and
gave only 1,4-disubstituted triazoles 3–12 in high yields. The con-
ditions, however, varied somewhat depending on the dipolarophi-
licity of the acetylene. Activated acetylenes, for example, methyl
propiolate, needed shorter time and lower temperature, while
the electron rich acetylenes required longer times, up to seven
days, and temperatures up to 45 °C. In only one case, with 4-meth-
oxyphenylacetylene, traces of 1,5-disubstituted regioisomer could
be detected. In the case of tosylacetylene, no reaction was observed
AcO
N
AcO
N3
1
HO
OH
OAc
O
OAc
O
N
N
N
a or b
c or d
O
SMe
SMe
SMe
N
N
R
R
AcO
AcO
HO
2 R=H (b, 81%)
16 R=H (c, 90%)
17 R=CO2Me (d, 90%)
18 R=(CH2)2CH3 (c, 77%)
19 R=CH2OH (c, 80%)
20 R=1-hydroxycyclohexyl (c, 87%)
21 R=Ph (c, 90%)
3 R=CO2Me (a, 95%)
4 R=(CH2)2CH3 (a, 99%)
5 R=CH2OH (a, 95%)
6 R=1-hydroxycyclohexyl (a, 96%)
7 R=Ph (a, 95%)
22 R=3-methoxyphenyl (c, 76%)
23 R=4-methoxyphenyl (c, 72%)
24 R=2-fluorophenyl (c, 82%)
25 R=1-naphthyl (c, 87%)
26 R=3-pyridyl (c, 81%)
8 R=3-methoxyphenyl (a, 76%)
9 R=4-methoxyphenyl (a, 74%)
10 R=2-fluorophenyl (a, 99%)
11 R=1-naphthyl (a, 84%)
12 R=3-pyridyl (a, 94%)
AcO
OAc
HO
OH
O
N
N
N
N
b
O
c
1
SMe
SMe
N
N
R1
R1
AcO
HO
R2
R2
27 R1=tosyl, R2=H (c, 75%)
13 R1=tosyl, R2=H (38%)
14 R1=H, R2=tosyl (52%)
28 R1=H, R2=tosyl (c, 75%)
29 R1=CONHMe, R2=CONHMe (c, 92%)
15 R1=CO2Me, R2=CO2Me (99%)
HO
OH
O
HO
OH
O
N
N
f
e
N
N
H
SMe
1
N
H2N
3
SMe
N
N
HO
R
HO
O
NH2
O
35 (60%)
30 R=Me (98%)
31 R=Bu (90%)
32 R=(CH2)3OH (86%)
33 R=(CH2)2 (75%)
34 R=Bn (80%)
N
O
Scheme 1. Reagents and conditions: (a) alkyne, CuI, DIPEA, toluene, rt–40 °C; (b) toluene, 65–80 °C; (c) 40% MeNH2 in H2O; (d) NaOMe, MeOH; (e) RNH2, MeOH; (f)
icyanoacetamide, K2CO3, DMSO, 50 °C, iiMeOH.

B. A. Salameh et al. / Bioorg. Med. Chem. 18 (2010) 5367–5378
5369
AcO
N3
OAc
O
AcO
N3
OAc
O
OAc
O
O
OMe
AcO
AcO OAc
42
AcO
NHAc
36
a
a
AcO
OAc
O
OAc
O
OAc
O
AcO
N
N N
N
N
N
O
N
O
R1
R2
R
R
OMe
AcO
MeO
AcO
NHAc
AcO
37 R=CO2Me (90%)
38 R=1-naphthyl (99%)
43 R1,R2=H, OAc (93%)
b
44 R1=H, R2=Br (82%)
HO
N
N
c
OH
O
OH
O
OAc
O
AcO
N
N N
O
O
OMe
HO
HO
S
NHAc
MeO
AcO
39 R=CONHMe (b, 80%)
40 R=CONHBn (b, 81%)
41 R=1-naphthyl (c, 75%)
2
45 (36%)
d
Scheme 2. Reagents and conditions: (a) alkyne, CuI, DIPEA, toluene; (b) RNH2,
MeOH; (c) 40% MeNH2 in H2O.
OH
O
HO
N
O
S
RHN
N N
HO
under the Cu(I) catalyzed conditions. Fortunately, both regioisom-
ers of the corresponding triazoles 13 and 14 could be obtained in
90% in a 2:3 ratio by heating 1 with tosylacetylene. The 1,4,5-tri-
substituted triazole 15 was obtained in a near quantitative yield
by reaction of the azide 1 with dimethyl acetylenedicarboxylate
under Huisgen thermal conditions.
2
46 R=Me (quant.)
47 R=Bu (quant.)
48 R=Bn (quant.)
49 R=(CH2)2Ph (quant.)
50 R=allyl (quant.)
The unprotected triazoles 16 and 18–29 were obtained by
treatment of 2 and 4–15 with aqueous methylamine, while the
methylester 17 was obtained by the treatment of 3 with methano-
lic sodium methoxide. Reaction of the triazole methyl ester 3 with
different amines gave a panel of amides 30–34. The amide forma-
tion was faster and proceeded at lower temperature for aliphatic
amines to give 30–33, whereas heating at 45 °C was necessary with
benzylamine to afford 34. Finally, an unprotected 1,4,5-trisubsti-
tuted triazole 35 was prepared with complete regioselectivity in
one step by treatment of 1 with cyanoacetamide.32
As LacNAc and thiodigalactoside are far superior to galactose as
natural ligands for galectins, three LacNAc and eight thiodigalacto-
side derivatives carrying triazoles at the galactose C30s were pre-
pared. Again, 4-carbamoyl and 4-aryl triazoles were targeted,
because these were earlier demonstrated to enhance affinity for
galectin-3 within a galactose series of compounds.29 The known
LacNAc C30-azido compound 3633 was somewhat less reactive than
the corresponding monosaccharide 1. Nevertheless, the corre-
sponding triazoles 37 and 38 were obtained in good yields
(Scheme 2). Treatment of 37 and 38 with methylamine and benzyl-
amine in methanol provided the methyl amide 39 and the benzyl
amide 40, respectively, while the 1-naphthyl triazole 41 was ob-
tained by aminolysis of the acetates with methylamine in water.
The thiodigalactoside triazole ester 45 was obtained starting
with cycloaddition between methyl propiolate and the known ga-
lacto azide 4234 (Scheme 3). The resulting triazole tetraacetate 43
was treated with HBr in AcOH to give the bromide 44. Dimerisation
of the bromide 44 was achieved by treatment with dried sodium
sulfide affording the di-triazole-substituted thiodigalactoside 45
in a moderate yield due to purification difficulties arising from
low solubility of 45. Deprotection of the common ester precursor
45 using a panel of primary amines in methanol, with concomitant
substitution of the methyl ester to give the respective amides, gave
a set of seven compounds 46–52.
51 R=(CH2)3OH (70%)
52 R=(CH2)2OMe (quant.)
Scheme 3. Reagents and conditions: (a) methyl propiolate, CuI, DIPEA, toluene; (b)
HBr (33% in AcOH), DCM, Ac2O; (c) Na2S (dried), MeCN, 4 Å; (e) RNH2, MeOH.
9N together with methyl b-D-galactoside 53, LacNAc methyl glyco-
side 54, and un-derivatized thiodigalactoside 55 (Chart 1) using a
competitive fluorescence polarization assay as earlier described
in detail35,36 (Table 1) and shown to correlate well to other protein
affinity assays, such as ITC and ELISA,20,22 and to in vitro and
HO
OH
O
HO
OMe
HO
53
HO
OH
O
OH
O
HO
O
OMe
HO
HO
NHAc
54
OH
O
HO
HO
HO
OH
S
O
HO
HO OH
55
HO
OH
O
H
OH
O
N
O
OMe
HO
HO
O
NHAc
56
HO
OH
O
O
MeO
OMe
HO
H
N
S
N
O
HO
H
HO
2.2. Affinity evaluation against galectin-3, 7, 8N, and 9N
MeO
OMe
O
OH
57
The triazoles 16–35, 39–41, and 46–52 were screened for inhi-
bition potency against the available human galectin-3, 7, 8N, and
Chart 1.

5370
B. A. Salameh et al. / Bioorg. Med. Chem. 18 (2010) 5367–5378
Table 1
Kd (l
M) valuesa for galactose triazoles 16–35, LacNAc triazoles 39–41, and thiodigalactoside triazoles 46–52 determined in a competitive fluorescence polarization assay35,36
Triazole substituents
Galectin
C4
Galactose derivatives
C5
3
7
8N
9N
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
–H
–H
–H
–H
–H
–H
–H
–H
–H
–H
–H
–H
–H
–Tosyl
–CONHMe
–H
>5000b
1300 180
>5000
2400 30
>5000
2200 1400
2500 170
>5000
>5000
>5000
>5000
>5000
>5000
>5000
>5000
ndc
>5000
>5000
670 290
nd
>5000
430 160
>5000
–CO2Me
–(CH2)CH3
–CH2OH
1-Hydroxycyclohexyl
Ph
3-Methoxyphenyl
4-Methoxyphenyl
2-Fluorophenyl
1-Naphthyl
3-Pyridyl
>5000
>5000
1600 260
150 11
130 10
260 60
200 38
120 26
1900 680
130 37
>5000
1000 310
1300 640
2100 1200
>5000
1700 760
690 360
1600 46
330 68
1600 1200
>5000
1100 500
640 39
1900 400
470 360
540 230
>5000
1700 700
ꢀ2800
ꢀ2000
>2000
810 380
nd
2000 200
nd
>5000
nd
1100 540
1500 800
>5000
2100 500
2700 1700
Tosyl
–H
–CONHMe
–CONHMe
nd
nd
>5000
230
4
>5000
>5000
>5000
>5000
>5000
>5000
–CONHBu
ꢁH
150 38
360 41
520 95
150 70
>2000
–CONH(CH2)3OH
–CONH(CH2)2N((CH2)2)2O
–CONHBn
–H
–H
–H
–NH2
–CONH2
LacNAc derivatives
39
40
41
–CONHMe
–CONHBn
1-Naphthyl
–H
–H
–H
3.8 1.1
1.5 0.3
0.66 0.09
130 76
74 25
83 11
>2000
>2000
>2000
73
12
13
6
4
8
Thiodigalactoside derivatives
46
47
48
49
50
51
52
–CONHMe
–CONHBu
–CONHBn
–CONHCH2CH2Ph
–CONHCH2CHCH2
–CONH(CH2)3OH
–CONH(CH2)2OMe
–H
–H
–H
–H
–H
–H
–H
0.12 0.08
0.029 0.07
0.39 0.17
0.15 0.02
0.065 0.040
0.12 0.02
0.25 0.15
8.0 1.6
5.4 0.9
8.3 2.1
2.0 0.4
3.4 0.2
5.0 0.3
4.7 1.3
81 27
0.91 0.06
1.1 0.3
0.53 0.22
1.1 0.4
0.74 0.17
2.1 0.4
2.3 1.6
58
47
20
5
1
5
66 22
58
9
90 19
References
53
54
55
56
57
Me b-
Me b-LacNAc
Thiodigalactoside
Me 30-naphth-2-amido-LacNAc
D
-galactopyranoside
440036,38
5939
480036
550039
16022
>100
530036
100039
6122
330036
49039
3822
4922
0.4820
0.05022
>100
>100
Di-(3,5-dimethoxybenzamido)-thiodigalactoside
2.822
ꢀ10022
1.822
Methyl b-D
-galactose 53, methyl b-LacNAc 54, and thiodigalactoside 55 are included as reference compounds, as are the amido-derivatized high-affinity inhibitors 30-naphth-
2-amido-LacNAc glycosides 5620 and thiodigalactoside di-(3,5-dimethoxy)-benzamide 57.22
a
Average and standard deviation of 4–16 single point measurements. Galectin-3, 8N, and 9N were measured at 20 °C and galectin-7 at 4 °C due to a lower assay sensitivity
for the latter.
b
Inhibitor concentrations were chosen to provide reliable Kd values up to 5 mM for monosaccharides 16-35 and 2 mM for disaccharides 39-41 and 46-52, as higher Kd
values are significantly above those for the corresponding reference compounds ligands and hence judged to be insignificant.
c
Not determined.
ex vivo evaluations.14,15,37 Furthermore, two aromatic amides were
included as reference to our previous work on galectin-3: the Lac-
NAc 2-naphthamide 5620 and the thiodigalactoside di-(3,5-dime-
thoxy)-benzamide 5722 were among the most potent galectin-3
inhibitors with Kd of 480 nM and 50 nM, respectively. The mono-
substituted triazole 16 was practically non-inhibitory for galec-
tin-3, 8N, and 9N, while the presence of a triazole ring was
tolerated by galectin-7. It is clear that the galactose C3-triazole is
unfavorable to recognition by the former three galectins. The inhi-
bition potency, however, was recovered for galectin-3 and to a les-
ser extent for galectin-7 and 9N, by the presence of triazole 4-
substitutents. Thus, although the presence of the triazole ring itself
decreases the inhibition activity, it positions the substituent at the
4-position to form favorable interactions with galectin-3, 7, and
9N.
the substitution pattern of the aromatic rings (21–25 and 27) ap-
peared to be less important, while introduction of a heteroatom,
that is, the 3-pyridyl substituted compound 26, decreased the
inhibitory power. Substitution at C5 of the triazole ring (28–29
and 35) was unfavorable to the interaction with galectin-3.
None of the galactosides 16–35 proved to be good inhibitors of
galectin-7 or 8N, with one exception for each of these galectins: the
1-naphthyl-substituted compound 25 was five times better than
the reference methyl galactoside 53 against galectin-7 and the 3-
pyridyl substituted compound 26 showed seven times higher affin-
ity than 53 for galectin-8N. The relatively good affinity of 26 for
galectin-8N is most likely a result of a specific interaction of the
pyridine nitrogen with galectin-8N, as the corresponding phenyl
substituted compound 21 is inactive against this galectin.
On the contrary, galectin-9N was inhibited well by several com-
pounds, albeit still with modest affinity enhancements relative to
53.
For galectin-3, the aliphatic substituted triazoles 18–20 showed
inhibition potency similar to the methyl galactoside 53, while aro-
matic (21–25 and 27) and carbamoyl (30–34) substituents at the
triazole 4-position resulted in particularly efficient inhibitors
approaching the LacNAc disaccharide 54 in affinity. Interestingly,
Attaching selected triazole groups onto the better natural galec-
tin ligand LacNAc to obtain two carbamoyl derivatives 39–40 and
one 1-naphthyl derivative 41 resulted in affinity enhancing effects

B. A. Salameh et al. / Bioorg. Med. Chem. 18 (2010) 5367–5378
5371
paralleling those observed for the above discussed triazol-1-yl
galactosides 25, 30, and 34. The 1-naphthyl compound 41 was par-
ticularly potent as inhibitor of galectin-3 with a Kd of 660 nM,
which in the range of the reported best LacNAc-based inhibitor
30-(2-naphthamido)-LacNAc 5620 (Kd 480 nM). Analogously, the
amides 39 and 40 were low
lM inhibitors of galectin-7 and 9N
and are as such among the best LacNAc-derived inhibitors reported
of these two galectins.
All the thiodigalactoside-derived amide-triazoles 46–52
showed large increases in binding to galectin-3 compared to
underivatised thiodigalactoside 55. In addition, the methylamide
46 affinity enhancement much greater than the analogous Lac-
NAc-derived compound 39 suggesting that both triazole-methyl
amide substituents of 46 interact favorably with galectin-3. The
best triazolyl thiodigalactoside, the bis-butylamido compound 47,
binds to galectin-3 with a Kd of 29 nM, which is quite remarkable
for a carbohydrate–lectin interaction and similar to that of the cor-
responding benzamides22 (e.g., 57). The variation between the dif-
ferent amides 46–52 was relatively low.
Against galectins-7 and -9N, the amides 46–52 once again
showed increases in affinity, but not as big as for galectin-3. In con-
trast, for galectin-8N very small increase for 46–52 or even loss in
affinity was observed. Hence, the di-triazolyl thiodigalactosides
46–55 display significant selectivity for galectin-3 over galectin-7,
8N, and9N, whichpoints toanimportantadvantagetheseinhibitors.
Examination of a sequence alignment of the different carbohy-
drate recognition domains reveals a possible explanation for the
high affinity of 46–52 for galectin-3, 7, and 9N. The increases in affin-
ity for these three galectin-3 for the thiodigalactoside di-amides
Figure 1. (a) Conformational analysis and NOE experiment of 21 show that it
adopts a conformation with the triazole C5 almost antiperiplanar to galactose H3.
This conformation is supported by ꢀ10% NOE from the triazole H5 to galactose H2
(depicted by a black arrow). (b) Conformational analysis and NOE experiment of 29
show it adopts a conformation with the triazole N2 almost antiperiplanar to
galactose H3. This conformation positions the triazole C5 substituent in a steric
clash with the galectin’s conserved tryptophan residue onto which galactose forms
a CH–p stacking interaction.
(e.g., 57) were due to cation–p interactions with two arginine side-
chains (Arg-144 and Arg-186).22 The analogous increases for 46–
52 could be due to similar double arginine interactions. In both
galectins-7 and -9N, both of these arginine residues are also present
in the vicinity of the carbohydrate binding site (Arg-31 and Arg-79
for galectin-7; Arg-44 and Arg-87 for galectin-9N). However, for
galectin-8, the arginine residue corresponding to Arg-186 in galec-
tin-3 is missing; instead there is an isoleucine residue.
3. Conclusions
In conclusion, we reported the synthesis of a novel class of
galectin inhibitors distinguished by their easy preparation, high
stability, and high activity that could be applied to monosaccharide
and disaccharides. The affinity of LacNAc C30-triazoles and thiodi-
galactoside C3,C30-di-triazoles reported herein equals that of the
corresponding high-affinity aromatic amido compounds, but have
an important advantage in that they are easier to synthesize. In
particular, the thiodigalactoside derivatives, such as 47, are prom-
ising for development of clinically useful galectin-3 inhibitors due
to their low nM affinity, high selectivity over galectin-7, 8N, and
9N, less complex synthesis, and expected improved hydrolytic sta-
bility relative to O-glycosidic inhibitors.
2.3. Conformational analysis and structure–activity
relationships
The three 5-substituted triazoles (28–29 and 35) were rather
poor inhibitors of the galectins. Conformational analysis (MMFFs
force field in water implemented in Macromodel 9.0) of the galact-
ose C3–triazole-N1 bond in compounds 16, 18, 27–30, and 35 all
showed two minimum conformations: One minimum with the tri-
azole C5 positioned between galactose H3 and O4 and the triazole
4. Experimental
4.1. General
N1–N3 placed on the
a-side of the galactose ring and a second
minimum with the triazole turned approximately 180° and the tri-
azole N2 positioned between galactose H3 and O4 (Fig. 1). The for-
mer conformation was favored with around 6 kJ/mol for
compounds lacking substituents at the triazole C5 (16 18, 21, 27,
and 30) and is possibly the bioactive conformation (Fig. 1a). Sup-
port that this conformation is significantly populated came from
the observation of strong NOE (>10%) enhancements between the
galactose H2 and the triazole H5 in compounds 21 and 27. How-
ever, the corresponding conformation was disfavored in the 5-
substituted triazoles 28–29 by 20–30 kJ/mol. Thus, compounds
28–29 predominantly populate conformations that project the tri-
azole C5-substituent into a position bound to interfere sterically
with the galectin surfaces, that is, to the conserved tryptophan res-
Melting points were recorded on a Kofler apparatus (Reichert)
and are uncorrected. 1H NMR spectra were recorded with Bruker
DRX-400 or ARX-300 instruments. Chemical shifts are reported rel-
ative to Me4Si and were calculated using the residual solvent peak
as a reference. Chemical shifts and coupling constants were ob-
tained from 1H NMR spectra and proton resonances were assigned
from COSY experiments. Low- and high-resolution (HRMS) fast
atom bombardment mass spectra were recorded using a JEOL SX-
120 instrument. Matrix-assisted laser desorption/ionization-time
of flight mass spectroscopy (MALDI-TOF MS) was carried out with
a Bruker Biflex III instrument with gentisic acid as matrix. Optical
rotations were measured on a Perkin–Elmer 341 polarimeter with
a path length of 1 dm; concentrations are given in g per 100 mL.
Thin layer chromatography (TLC) was carried out on Merck Kiesel-
gel sheets, pre-coated with 60F254 silica. Plates were developed
using 10% sulfuric acid. Column Chromatography was performed
idue that stacks with the b-galactoside a-face of ligands (Fig. 1b).
The compound carrying a smaller amino-substituent at triazole
C5 (35) displayed similar energy for both minima and conse-
quently showed intermediate inhibitory power for galectin-3, 7,
and 9N.

5372
B. A. Salameh et al. / Bioorg. Med. Chem. 18 (2010) 5367–5378
on SiO2 (Matrex, 60 Å, 35 70
l
m, Grace Amicon) and thin layer
4.2.3. Methyl 2,4,6-tri-O-acetyl-3-deoxy-3-(4-propyl-1H-[1,2,3]-
triazol-1-yl)-1-thio-b- -galactopyranoside 4
chromatography (TLC) was carried out on 60F254 silica (Merck).
Solutions were concentrated by using rotary evaporation with a
bath temperature at or below 40 °C. Dichloromethane was dis-
tilled from calcium hydride immediately before use. Acetonitrile
was distilled from calcium hydride and stored over 4 Å molecular
sieves. All other reagents and solvents were used as supplied.
Affinities of compounds for galectins were determined in a com-
petitive fluorescence polarization assay as described previ-
ously.35,36 Molecular modeling was performed with the MMFFs
force field with water implemented in MacroModel (version 9.1,
Schrödinger, LLC, New York, 2005). Starting conformations were
built from the published crystal structure of galectin-3 in complex
D
Method A, x = 1-pentyne, t = three days, T = 50 °C, column elu-
ent heptane–ethyl acetate 5:2. Yield 11.9 mg, 99%. 1H NMR
(400 MHz, CDCl3) d 7.31 (s, 1H, H-50), 5.66 (dd, 1H, J2,3 = 11.1,
H-2), 5.55 (d, 1H, H-4), 5.12 (dd, 1H, J3,4 = 3.2, H-3), 4.53 (d, 1H,
J1,2 = 9.5, H-1), 2.65 (td, 2H, JH,H = 7.2, JH,H = 1.4, CH2Ar), 2.24, 2.05,
2.04, 1.91 (4s, each 3H, 4CH3), 1.69–1.57 (m, 2H, CH2), 0.89 (t,
3H, JH,H = 7.3, CH3CH2). 13C NMR (100.6 MHz, CDCl3) d 170.3,
169.5, 168.5 (3C@O), 148.3 (C-40), 119.1 (C-50), 84.2 (C-19,) 75.3
(C-5), 68.8 (C-4), 65,5 (C-2), 62.6 (C-3), 61.3 (C-6), 27.4 (CH2Ar),
22.6 (CH2), 20.5, 20.4, 20.3 (3CH3C@O), 13.3 (CH2), 11.5 (CH3S).
MALDI-TOF MS for C18H28N3O7S [M+H]+ 430.
with
a
C30-amido-derivatised LacNAc-based inhibitor,20 The
minimizations converged in all cases and the binding modes and
overall structures of the minimized complexes closely resembled
that of the crystal structures used for building starting conforma-
tions.
4.2.4. Methyl 2,4,6-tri-O-acetyl-3-deoxy-3-(4-hydroxymethyl-1H
-[1,2,3]-triazol-1-yl)-1-thio-b-D-galactopyranoside 5
Method A, x = propargyl alcohol, t = 12 h, T = rt, column eluent
heptane–ethyl acetate 3:1. Yield 11.0 mg, 95%. 1H NMR
(400 MHz, CDCl3) d 7.60 (s, 1H, H-50), 5.70 (dd, 1H, J2,3 = 11.0,
H-2), 5.54 (d, 1H, J3,4 = 3.0, H-4), 5.14 (br d, 1H, H-3), 4.80–4.70
(m, 2H, CH2OH), 4.55 (d, 1H, J1,2 = 9.5, H-1), 4.14 (s, 3H, H-5,
2H-6), 2.25, 2.06, 2.04, 1.93 (4s, each 3H, 4CH3). 13C NMR
(100.6 MHz, CDCl3) d 170.3, 169.5, 168.8 (3C@O), 147.9 (C-40),
120.4 (C-50), 84.1 (C-1), 75.3 (C-5), 68.7 (C-4), 65.4 (C-2), 62.8
(C-3), 61.4 (C-6), 56.4 (CH2), 20.6, 20.5, 20.3 (3CH3C@O), 11.4
(CH3S). MALDI-TOF MS [M+H]+ 418, [M+Na]+ 440.
4.2. General procedures for the preparation of triazoles 2–15
and 37–38
Method A for compounds 3–12 and 37–38: A mixture of methyl
2,4,6-tri-O-acetyl-3-azido-3-deoxy-1-thio-b-
120 (10 mg, 0.028 mmol) for 3–12 or methyl 2,4,6-tri-O-acetyl-3-
azido-3-deoxy-b- -galactopyranosyl-(1–4)-2-acetamido-3,6-di-O-
acetyl-2-deoxy-b-
-glucopyranoside 3633 (10 mg, 0.0158 mmol)
D-galactopyranoside
D
D
4.2.5. Methyl 2,4,6-tri-O-acetyl-3-deoxy-3-(4-(1-hydroxycyclo-
hexyl)-1H-[1,2,3]-triazol-1-yl)-1-thio-b-D-galactopyranoside 6
for 37 and 38, the acetylene derivative (x, 1 equiv), CuI (0.5 mg,
0.1 equiv), diisopropylethylamine (1 equiv) and toluene (1 mL)
was stirred together for (t) time at (T) temperature. The solvent
was evaporated and the product was purified by column chroma-
tography using the eluent indicated in each case.
Method A, x = 1-ethynyl-1-cyclohexanol, t = 12 h, T = rt, column
eluent heptane–ethyl acetate 3:2. Yield 12.0 mg, 96%. 1H NMR
(400 MHz, CDCl3) d 7.49 (s, 1H, H-50), 5.67 (dd, 1H, J2,3 = 11.0,
H-2), 5.54 (d, 1H, H-4), 5.13 (dd, 1H, J3,4 = 3.2, H-3), 4.55 (d, 1H,
J1,2 = 9.6, H-1), 4.13 (s, 3H, H-5, 2H-6), 2.24, 2.05, 2.04, 1.91 (4s,
each 3H, 4CH3), 1.81–1.25 (m, 11H, cyclohexyl). 13C NMR
(100.6 MHz, CDCl3) d 170.3, 169.4, 168.6 (3C@O), 155.7 (C-40),
118.4 (C-50), 84.1 (C-1), 75.3 (C-5), 69.4 (cyclohexyl C-1), 68.8
(C-4), 65.5 (C-2), 62.7 (C-3), 61.3 (C-6), 38.1, 38.0, 25.2, 21.9, 21.8
(cyclohexyl), 20.5, 20.4, 20.3 (3CH3C@O), 11.5 (CH3S). MALDI-TOF
MS for C21H32N3O8S [M+H]+ 486.
Method B for compounds 2 and 13–15: A mixture of methyl
2,4,6-tri-O-acetyl-3-azido-3-deoxy-1-thio-b-D
-galactopyranoside19
(10 mg, 0.028 mmol), the corresponding acetylene derivative
(x, 4 equiv) and toluene (1.5 mL) was heated at (T) temperature
for 12 h. After evaporation of the solvent, the product was purified
by column chromatography using the eluent indicated in each
case.
4.2.1. Methyl 2,4,6-tri-i-acetyl-3-(1H-[1,2,3]-triazol-1-yl)-3-
deoxy-1-thio-b-D-galactopyranoside 2
4.2.6. Methyl 2,4,6-tri-O-acetyl-3-deoxy-3-(4-phenyl-1H-[1,2,3]-
triazol-1-yl)-1-thio-b-D-galactopyranoside 7
Method B, x = propiolic acid, t = 17 h, T = 80 °C, column eluent
heptane–ethyl acetate 3:2. Yield 8.7 mg, 81%. 1H NMR (400 MHz,
CDCl3) d 7.67 (br d, 1H, JH,H = 0.8, triazole), 7.61 (br d, 1H, triazole),
5.71 (dd, 1H, J2,3 = 11.0, H-2), 5.57 (d, 1H, H-4), 5.19 (dd, 1H,
J3,4 = 3.2, H-3), 4.56 (d, 1H, J1,2 = 9.5, H-1), 4.14 (s, 3H, H-5, 2H-6),
2.25, 2.05, 2.04, 1.91 (4s, each 3H, 4CH3). 13C NMR (100.6 MHz,
CDCl3) d 170.3, 169.5, 168.6 (3C@O), 133.8, 122.0 (C-40, C-50),
84.1 (C-1), 75.3 (C-5), 68.7 (C-4), 65.4 (C-2), 62.7 (C-3), 61.3 (C-
6), 20.6, 20.4, 20.3 (3CH3C@O), 11.5 (CH3S). MALDI-TOF MS for
Method A, x = phenylacetylene, t = three days, T = rt, column
eluent heptane–ethyl acetate 5:2. Yield 12.1 mg, 95%. 1H NMR
(400 MHz, CDCl3) d 7.82 (s, 1H, H-50), 7.80–7.78 (m, 2H, o-Ph),
7.44–7.40 (m, 2H, p-Ph), 7.34 (tt, 1H, Jo,m = 7.3, Jo,p = 1.1, p-Ph),
5.76 (dd, 1H, J2,3 = 11.0, H-2), 5.62 (d, 1H, H-4), 5.19 (dd, 1H,
J3,4 = 3.2, H-3), 4.57 (d, 1H, J1,2 = 9.5, H-1), 4.15 (s, 3H, H-4),
2.27, 2.06, 2.05, 1.93 (4s, each 3H, 3CH3). 13C NMR (100.6 MHz,
CDCl3) d 170.3, 169.6, 168.6 (3C@O), 147.8 (C-40), 129.9, 128.8
[2C], 128.3, 128.6 [2C] (Ph), 117.9 (C-50), 84.1 (C-1), 75.4 (C-5),
68.8 (C-4), 65.4 (C-2), 62.9 (C-3), 61.4 (C-6), 20.6, 20.4, 20.3
(3CH3C@O), 11.5 (CH3S). MALDI-TOF MS for C21H26N3O7S
[M+H]+ 464.
C
15H22N3O7S [M+H]+ 388.
4.2.2. Methyl 2,4,6-tri-O-acetyl-3-deoxy-3-(4-methoxycarbonyl-
1H-[1,2,3]-triazol-1-yl)-1-thio-b- -galactopyranoside 3
D
Method A, x = methyl propiolate, t = 12 h, T = rt, column eluent
heptane–ethyl acetate 3:2. Yield 11.6 mg, 95%. 1H NMR (400 MHz,
CDCl3) d 8.14 (s, 1H, H-50), 5.70 (dd, 1H, J1,2 = 9.5, H-1), 5.57 (d, 1H,
J3,4 = 3.2, H-4), 5.18 (dd, 1H, H-3), 4.55 (d, 1H, J1,2 = 9.5, H-1), 4.14
(m, 3H, H-5, 2H-6), 3.93 (s, 3H, CH3O), 2.25, 2.08, 2.04, 1.92 (4s, each
3H, 4CH3). 13C NMR (100.6 MHz, CDCl3) d 170.7, 169.9, 169.2, 161.0
(4C = O), 140.6 (C-40), 126.8 (C-50), 84.6 (C-1), 75.8 (C-5), 68.9 (C-4),
65.9 (C-2), 63.8 (C-3), 61.7 (C-6), 52.7 (CH3O), 21.0, 20.9, 20.8
(3CH3C@O), 11.9 (CH3S). MALDI-TOF MS for C17H24N3O9S [M+H]+
446.
4.2.7. Methyl 2,4,6-tri-O-acetyl-3-deoxy-3-(4-(3-methoxyphenyl)-
1H-[1,2,3]-triazol-1-yl)-1-thio-b-D-galactopyranoside 8
Method A, x = 1-ethynyl-3-methoxybenzene, t = seven days,
T = 40 °C, column eluent heptane–ethyl acetate 5:2. Yield
1
10.4 mg, 76%. H NMR (400 MHz, CDCl3) d 7.87 (s, 1H, H-50),
7.42–7.30 (m, 3H, Ar), 6.91–6.88 (td, 1H, JH,H = 2.4, 7.0, Ar), 5.75
(dd, 1H, J2,3 = 11.0, H-2), 5.62 (d, 1H, J3,4 = 3.1, H-4), 5.18 (dd, 1H,
H-3), 4.57 (d, 1H, J1,2 = 9.5, H-1), 4.15 (s, 3H, H-5, 2H-6), 3.87 (s,
3H, CH3O), 2.26, 1.93 (2s, each 3H, 2CH3), 2.06 (s, 6H, 2CH3). 13C
NMR (100.6 MHz, CDCl3) d 170.3, 169.6, 168.6 (3C@O), 160.0,

B. A. Salameh et al. / Bioorg. Med. Chem. 18 (2010) 5367–5378
5373
147.7, 131.2 (C-40, C1-Ph, C3-Ph), 129.8, 118.0, 114.4, 110.8 (Ph),
118.2 (C-50), 84.2 (C-1), 75.4 (C-5), 68.8 (C-4), 65.4 (C-2), 62.9
(C-3), 61.4 (C-6), 55.3 (CH3O), 20.6, 20.4, 20.3 (3CH3C@O), 11.5
(CH3S). MALDI-TOF MS [M+H]+ = 494, [M+Na]+ = 516.
4.2.12. Methyl 2,4,6-tri-O-acetyl-3-(4-p-tolylsulfonyl-1H-[1,2,3]-
triazol-1-yl)-3-deoxy-1-thio-b- -galactopyranoside 13 and methyl
2,4,6-tri-O-acetyl-3-(5-p-tolylsulfonyl-1H-[1,2,3]-triazol-1-yl)-
3-deoxy-1-thio-b- -galactopyranoside 14
D
D
Method B, x = ethynyl-p-tolylsulfone, t = 20 h, T = 65 °C, column
eluent heptane–ethyl acetate 5:1–7:2 gradient, gave 13 (5.7 mg,
38%) and 14 (7.8 mg, 52%). 1H NMR for 13 (400 MHz, CDCl3) d
8.17 (s, 1H, H-50), 7.88 (d, 2H, JH,H = 8.4, Ph), 7.33 (d, 2H, Ph), 5.64
(dd, 1H, J2,3 = 11.0, H-2), 5.51 (d, 1H, H-4), 5.15 (dd, 1H, J3,4 = 3.2,
H-3), 4.53 (d, 1H, J1,2 = 9.5, H-1), 4.12 (m, 3H, H-5, 2H-6), 2.42,
2.23, 2.03, 2.00, 1.87 (5s, each 3H, 5CH3). 13C NMR for 13
(100.6 MHz, CDCl3) d 170.2, 169.3, 168.6 (3C@O), 149.5, 145.0,
139.9 (C-40, Ph C-1, Ph C-4), 129.8, 127.9 (each 2C, Ph), 124.9
(C-50), 83.9 (C-1), 75.2 (C-5), 68.3 (C-4), 65.2 (C-2), 63.5 (C-3),
61.1 (C-6), 21.6, 20.5, 20.3, 20.1 (3CH3C@O, CH3Ph), 11.4 (CH3S).
FAB-HRMS calcd for 13 C22H27O9N3NaS2 [M+Na]+ 564.1086; found
564.1103. 1H NMR for 14 (400 MHz, CDCl3) d 7.88 (d, 2H, JH,H = 8.3,
o-Ph), 7.87 (s, 1H, H-40), 7.43 (d, 2H, m-Ph), 6.06 (t, 1H, H-2), 5.43
(dd, 1H, J2,3 = 10.7, H-3), 5.25 (d, 1H, J3,4 = 2.6, H-4), 4.50 (d, 1H,
J1,2 = 9.7, H-1), 4.19–4.08 (m, 3H, H-5, 2H-6), 2.47, 2.25, 2.05,
1.99, 1.73 (5s, each 3H, 5CH3). 13C NMR for 14 (100.6 MHz, CDCl3)
d 170.3, 169.5, 168.3 (3C@O), 146.5, 138.2, 136.3 (Ph C-1, Ph C-4,
C-50), 136.8 (C-40), 130.3, 127.9 (each 2C, Ph), 84.1 (C-1), 75.1
(C-5), 67.6 (C-4), 65.3 (C-2), 62.4 (C-3), 61.6 (C-6), 21.7, 20.6,
20.3, 20.2, 11.1 (5CH3). FAB-HRMS calcd for 14 C22H27O9N3NaS2
[M+Na]+ 564.1086; found 564.1081.
4.2.8. Methyl 2,4,6-tri-O-acetyl-3-deoxy-3-(4-(4-methoxyphenyl)-
1H-[1,2,3]-triazol-1-yl)-1-thio-b-D-galactopyranoside 9
Method A, x = 1-ethynyl-4-methoxybenzene, t = seven days,
T = 40 °C, column eluent heptane–ethyl acetate 5:2. Yield
1
10.1 mg, 74%. H NMR (400 MHz, CDCl3) d 7.72 (s, 1H, H-50), 7.71
(m, 2H, m-Ph), 6.95 (m, 2H, o-Ph), 5.75 (dd, 1H, H-2), 5.62 (d, 1H,
J1,2 = 9.5, H-4), 5.17 (dd, 1H, J2,3 = 11.1, H-3), 4.56 (d, 1H, J1,2 = 9.5,
H-1), 4.15 (s, 3H, H-5, 2H-6), 3.84 (s, 3H, CH3O), 2.27, 1.96 (2s, each
3H, 2CH3), 2.05 (s, 6H, 2CH3). 13C NMR (100.6 MHz, CDCl3) d 170.3,
169.6, 168.6 (3C@O), 159.7, 147.7, 122.6 (C-40, C1-Ph, C4-Ph), 127.0
[2C], 114.2 [2C] (Ph), 117.1 (C-50), 84.2 (C-1), 75.4 (C-5), 68.8 (C-4),
65.4 (C-2), 62.8 (C-3), 61.4 (C-6), 55.7 (CH3O), 20.6, 20.5, 20.3
(3CH3C@O), 11.5 (CH3S). MALDI-TOF MS [M+H]+ = 494, [M+Na]+ =
516.
4.2.9. Methyl 2,4,6-tri-O-acetyl-3-deoxy-3-(4-(2-fluorophenyl)-
1H-[1,2,3]-triazol-1-yl)-1-thio-b-D-galactopyranoside 10
Method A, x = 1-ethynyl-2-fluorobenzene, t = 12 h, T = 40 °C,
column eluent heptane–ethyl acetate 5:2. Yield 13.3 mg, 99%. 1H
NMR (400 MHz, CDCl3) d 8.25 (dt, 1H, J = 1.8, 7.8, Ar), 7.98 (d, 1H,
JH,F = 3.5, H-50), 7.31 (m, 1H, Ar), 7.27–7.12 (m, 2H, Ar), 7.13 (ddd,
1H, J = 1.1, 8.2, 9.3, Ar), 5.75 (dd, 1H, H-2), 5.61 (d, 1H, J3,4 = 3.2,
H-4), 5.20 (dd, 1H, J2,3 = 11.1, H-3), 4.57 (d, 1H, J1,2 = 9.6, H-1),
4.16 (s, 3H, H-5, 2H-6), 2.27, 2.10, 2.05, 1.92 (4s, each 3H, 4CH3).
13C NMR (100.6 MHz, CDCl3) d 170.3, 169.4, 168.8 (3C@O), 159.2
(d, JC–F = 248, C2-Ph), 141.3 (d, JC–F = 2.3, C-40), 129.5 (d, JC–F = 8.5,
Ar), 127.8 (d, JC–F = 3.4, Ar), 124.5 (d, JC–F = 3.2, Ar), 121.2 (d,
JC–F = 13, C-50), 118.0 (d, JC–F = 13, C1-Ph,), 115.6 (d, JC–F = 22, Ar),
84.2 (C-1), 75.4 (C-5), 68.6 (C-4), 65.5 (C-2), 62.9 (C-3), 61.4
(C-6), 20.6, 20.4, 20.2 (3CH3C@O), 11.5 (CH3S). MALDI-TOF MS
[M+H]+ = 482, [M+Na]+ = 504.
4.2.13. Methyl 2,4,6-tri-O-acetyl-3-(4,5-bis-methoxycarbonyl-
1H-[1,2,3]-triazol-1-yl)-3-deoxy-1-thio-b-D-galactopyranoside
15
Method B, x = dimethyl acetylenedicarboxylate, T = 80 °C, 12 h,
column eluent heptane–ethyl acetate 1:1. Yield 13.9 mg, 99%. 1H
NMR (400 MHz, CDCl3) d 6.12 (dd, 1H, J2,3 = 10.6, H-2), 5.59 (d,
1H, H-4), 5.45 (dd, 1H, J3,4 = 2.9, H-3), 4.49 (d, 1H, J1,2 = 9.7, H-1),
4.24–4.14 (m, 3H, H-5, 2H-6), 4.02 (s, 3H, CH3O), 2.26, 2.03, 2.01,
1.87 (each s, each 3H, 4CH3). 13C NMR (100.6 MHz, CDCl3) d
170.3, 169.7, 168.3, 160.2, 158.9 (5C@O), 140.5, 129.5 (C-40, C-50),
84.2 (C-1), 75.1 (C-5), 67.7 (C-4), 65.3 (C-2), 62.8 (C-3), 61.5 (C-
6), 53.5, 52.7 (2CH3O), 20.6, 20.4, 20.2 (3CH3C@O), 11.1 (CH3S).
MALDI-TOF MS [M+H]+ 504, [M+Na]+ 526.
4.2.10. Methyl 2,4,6-tri-O-acetyl-3-deoxy-3-(4-(1-naphthyl)-1H-
[1,2,3]-triazol-1-yl)-1-thio-b-D-galactopyranoside 11
Method A, x = 1-ethynylnaphthalene, t = five days, T = 40 °C, col-
umn eluent heptane–ethyl acetate 5:2. Yield 11.9 mg, 84%, 1H NMR
(400 MHz, CDCl3) d 8.20–8.17 (m, 1H, Ar), 7.92–7.89 (m, 2H, Ar),
7.87 (s, 1H, H-50), 7.65 (dd, 1H, JH,H = 7.2, 1.1, Ar), 7.55–7.50 (m,
3H, Ar), 5.80 (dd, 1H, H-2), 5.71 (d, 1H, H-4), 5.28 (dd, 1H,
J2,3 = 11.1, J3,4 = 3.2, H-3), 4.61 (d, 1H, J1,2 = 9.5, H-1), 4.19–4.17
(m, 3H, H-5, 2H-6), 2.23, 2.07, 2.06, 1.99 (4s, each 3H, 4CH3). 13C
NMR (100.6 MHz, CDCl3) d 170.3, 169.6, 168.7 (3C@O), 146.9,
133.7, 131.1, 129.1, 128.4, 127.4, 127.3, 126.7, 125.9, 125.2, 125.0
(C-40, naphthalene), 121.1 (C-50), 84.2 (C-1), 75.4 (C-5), 68.8 (C-
4), 65.6 (C-2), 63.0 (C-3), 61.3 (C-6), 20.6, 20.5, 20.4 (3CH3C@O),
11.5 (CH3S). MALDI-TOF MS [M+H]+ = 514.
4.2.14. Methyl 2,4,6-tri-O-acetyl-3-deoxy-3-(4-methoxycarbonyl-
1H-[1,2,3]-triazol-1-yl)-b-D-galactopyranosyl-(1–4)-2-acetamido-
3,6-di-O-acetyl-2-deoxy-b-D-glucopyranoside 37
Method A, x = methyl propiolate, t = 24 h, T = 45 °C, column elu-
ent toluene–acetone 2:1. Yield 10.1 mg, 90%. 1H NMR (400 MHz,
CDCl3) d 8.14 (s, 1H, H-500), 5.64 (d, 1H, J2,NH = 9.4, NH), 5.55 (dd,
0
0
1H, J02 = 11.5, H-20), 5.49 (d, 1H, J03 = 3.2, H-40), 5.16 (dd, 1H,
,3
,4
H-30), 5.12 (dd, 1H, J2,3 = 9.7, H-3), 4.68 (d, 1H, J1 ,2 = 7.6, H-10), 4.49
(dd, 1H, J5,6a = 2.6, JH,H = 11.9, H-6a), 4.40 (d, 1H, J1,2 = 7.7, H-1),
4.16 (dd, 1H, J5,6b = 5.4, H-6b), 4.10 (s, 3H, H-50, 2H-60), 4.03 (dt, 1H,
H-2), 3.93 (s, 3H, CH3O), 3.83 (t, 1H, J = 8.7, H-4), 3.65 (ddd, 1H, H-
5), 3.46 (s, 3H, CH3O), 2.14 (s, 3H, CH3), 2.08 (s, 6H, 2CH3), 2.05,
1.98, 1.89 (each s, each 3H, 3CH3C@O). 13C NMR (100.6 MHz, CDCl3)
d 170.6, 170.3, 170.2, 170.1, 169.0, 168.7, 160.5 (7C@O), 140.1 (C-400),
126.7 (C-500), 101.7 (C-1), 101.0 (C-10), 75.7 (C-40), 72.5 (C-5), 72.1
(C-3), 71.7 (C-50), 67.8 [2C] (C-20, C-40), 62.1 (C-30), 62.0 (C-6), 60.7
(C-60), 56.6 (CH3O), 53.2 (C-2), 52.2 (CH3O), 23.2, 20.7 [2C], 20.5,
320.2, 20.1 (6CH3). FAB-HRMS calcd for C29H41N4O17 [M+H]+
717.2467; found, 717.2457.
0
0
4.2.11. Methyl 2,4,6-tri-O-acetyl-3-deoxy-3-(4-(3-pyridyl)-1H-
[1,2,3]-triazol-1-yl)-1-thio-b-D-galactopyranoside 12
Method A, x = 3-ethynyl pyridine, t = two days, T = 40 °C, col-
umn eluent toluene–acetone 3:1. Yield 12.0 mg, 94%, 1H NMR
(400 MHz, CDCl3) d 8.23 (br d, 1H, J = 6.8, Ar), 7.92 (s, 1H, H-50),
7.80–7.25 (bs, 1H, Ar), 7.27–7.23 (m, 1H, Ar), 7.18–7.14 (m, 1H,
Ar), 5.78 (dd, 1H, H-2), 5.62 (d, 1H, J3,4 = 3.1, H-4), 5.20 (dd, 1H,
J2,3 = 11.1, H-3), 4.58 (d, 1H, J1,2 = 9.5, H-1), 2.27, 2.08, 2.06, 1.94
(4s, each 3H, 4CH3). 13C NMR (100.6 MHz, CDCl3) d = 170.3,
169.5, 168.7 (3C@O), 133.0, 128.9, 128.1, 125.2 (C-40, pyridine),
118.7 (C-50), 84.1 (C-1), 75.3 (C-5), 68.7 (C-4), 65.5 (C-2), 63.1
(C-3), 61.3 (C-6), 20.6, 20.5, 20.4 (3CH3C@O), 11.4 (CH3S). MALDI-
TOF MS [M+H]+ = 465, [M+Na]+ = 487.
4.2.15. Methyl 2,4,6-tri-O-acetyl-3-deoxy-3-(4-(1-naphthyl)-1H-
[1,2,3]-triazol-1-yl)-b-
deoxy-3,6-di-O-acetyl-b-
D
-galactopyranosyl-(1–4)-2-acetamido-2-
-glucopyranoside 38
D
Method A, x = 1-ethynylnaphthalene, t = 24 h, T = 45 °C, column
eluent toluene–acetone 3:1. Yield 12.4 mg, 99%. 1H NMR (400 MHz,

5374
B. A. Salameh et al. / Bioorg. Med. Chem. 18 (2010) 5367–5378
CDCl3) d 8.17 (m, 1H, Ar), 7.89 (m, 2H, Ar), 7.84 (s, 1H, H-500), 7.64
(dd, 1H, JH,H = 1.0, 7.1, Ar), 7.53–7.49 (m, 3H, Ar), 5.68–5.62 (m, 3H,
(dd, 1H, H-2), 4.08 (d, 1H, H-4), 3.82–3.53 (m, 3H, H-5, 2H-6),
2.25 (s, 3H, CH3S), 2.02–1.28 (m, 10H, cyclohexyl). 13C NMR
(75 MHz, CD3OD) d 156.3 (C-40), 121.9 (C-50), 88.7 (C-1), 81.0
(C-5), 70.4 (cyclohexyl), 69.8 (C-4), 68.8 (C-3), 67.7 (C-2), 62.3
(C-6), 38.9 [2C], 26.6, 23.1 [2C] (cyclohexyl), 12.1 (CH3S). FAB-HRMS
calcd for C15H26N3O5S [M+H]+ 360.1593; found, 360.1596.
NH, H-40, H-20), 5.24 (dd, 1H, J2 ,3 = 11.5, J3 ,4 = 3.2, H-30), 5.14 (dd,
0
0
0
0
1H, J2,3 = 9.8, J3,4 = 8.4, H-3), 4.73 (d, 1H, J1 ,2 = 7.6, H-10), 4.53 (dd,
1H, J6a,6b = 11.9, J5,6a = 2.6, H-6a), 4.40 (d, 1H, J1,2 = 7.7, H-1), 4.18
(dd, 1H, J5,6b = 5.2, H-6b), 4.14 (m, 3H, H-50, 2H-60), 4.04 (q, 1H,
JH,H = 9.5, H-2), 3.86 (t, 1H, JH,H = 8.7, H-4), 3.68–3.64 (m, 1H,
H-5), 3.46 (s, 3H, CH3O), 2.14, 2.10, 2.07, 2.05, 1.97, 1.95 (6s, each
3H, 6CH3). 13C NMR (100.6 MHz, CDCl3) d 170.6, 170.4, 170.3,
170.1, 169.2, 168.7 (6C@O), 146.9, 133.7, 131.0, 129.1, 128.4,
127.3, 126.7, 126.0, 125.2, 124.9 (C-400, naphthalene), 121.5
(C-500), 101.7 (C-1), 101.2 (C-10), 75.8 (C-4), 72.6 (C-5), 72.2 (C-3),
71.9 (C-50), 68.2 (C-40), 68.0 (C-20), 62.1, 61.9 (C-30, C-6), 60.8
(C-60), 56.6 (CH3O), 53.3 (C-2), 23.2, 20.8 [2C], 20.5, 20.4, 20.3
(CH3C@O). FAB-HRMS calcd for C37H44N4NaO15 [M+Na]+
807.2701; found 807.2697.
0
0
4.3.5. Methyl 3-deoxy-3-(4-phenyl-1H-[1,2,3]-triazol-1-yl)-1-
thio-b-D-galactopyranoside 21
Compound 7 gave 21 (column eluent heptane–ethyl acetate 1:3,
6.5 mg, 90%). 1H NMR (400 MHz, CD3OD) d 7.84–7.81 (m, 2H, o-Ph),
7.45–7.41 (m, 2H, m-Ph), 7.33 (tt, 1H, Jm,p = 7.4, Jo,p = 1.2, p-Ph), 4.83
(dd, 1H, J3,4 = 3.0, H-3), 4.49 (d, 1H, J1,2 = 9.4, H-1), 4.27 (dd, 1H,
J2,3 = 10.5, H-2), 4.14 (d, 1H, H-4), 3.82–3.68 (m, 3H, H-5, 2H-6),
13
2.27 (s, 3H, CH3S). C NMR (100.6 MHz, CD3OD) d 148.3 (C-40),
131.9, 130.0 [2C], 129.2 [2C], 126.6 (Ph), 121.8 (C-50), 88.7 (C-1),
81.0 (C-5), 69.8 (C-4), 69.1 (C-3), 67.7 (C-2), 62.4 (C-6), 12.1
(CH3S). FAB-HRMS calcd for C15H20N3O4S [M+H]+ 338.1174; found,
338.1179.
4.3. General procedure for de-O-acetylation of compounds 2
and 4–14 to give 16 and 18–28
The protected sugar (10 mg) was dissolved in methylamine
(40% solution in water, 2 mL) and stirred overnight. The mixture
was concentrated and the product was purified by column
chromatography.
4.3.6. Methyl 3-deoxy-3-(4-(3-methoxyphenyl)-1H-[1,2,3]-
triazol-1-yl)-1-thio-b-D-galactopyranoside 22
Compound 8 gave 22 (C18 RP-HPLC eluent H2O–acetonitrile
gradient, 5.6 mg, 75%). 1H NMR (400 MHz, CD3OD) d 8.41 (s, 1H,
H-50), 7.42–7.33 (m, 3H, Ar), 6.91–6.88 (m, 1H, Ar), 4.81 (dd, 1H,
H-3), 4.49 (d, 1H, J1,2 = 9.4, H-1), 4.27 (dd, 1H, J2,3 = 10.4, H-2),
4.13 (d, 1H, J3,4 = 2.8, H-4), 3.84 (s, 3H, CH3O), 3.81–3.69 (m, 3H,
H-5, 2H-6), 2.27 (s, 3H, CH3S). 13C NMR (100.6 MHz, CD3OD) d
161.7, 148.2, 133.2 (C-3-Ph, C-1-Ph, C-40), 131.0, 119.0, 115.0,
111.9 (Ph), 122.0 (C-50), 88.7 (C-1), 81.0 (C-5), 69.8 (C-4), 69.1
(C-3), 67.7 (C-2), 62.4 (C-6), 55.7 (CH3O), 12.1 (CH3S). FAB-HRMS
calcd for C16H21N3NaO5S [M+Na]+ 390.1100; found 390.1100.
4.3.1. Methyl 3-deoxy-3-(1H-[1,2,3]-triazol-1-yl)-1-thio-b-D-
galactopyranoside 16
Compound 2 gave 16 (column eluent CH2Cl2–MeOH 17:1,
6.0 mg, 90%). 1H NMR (400 MHz, CD3OD) d 8.08 (d, 1H, JH,H = 1.0,
triazole), 7.74 (d, 1H, triazole), 4.83 (dd, 1H, J3,4 = 3.0, J2,3 = 10.5,
H-3), 4.46 (d, 1H, J1,2 = 9.4, H-1), 4.20 (dd, 1H, H-2), 4.09 (d, 1H,
H-4), 3.80–3.66 (m, 3H, H-5, 2H-6), 2.25 (s, 3H, CH3S). 13C NMR
(100.6 MHz, CD3OD) d 133.8, 125.4 (C-40, C-50), 88.7 (C-1), 81.0
(C-5), 69.8 (C-4), 68.8 (C-3), 67.7 (C-2), 62.4 (C-6), 12.6 (CH3S).
FAB-HRMS calcd for C9H15N3NaO4S [M+Na]+ 284.0681; found
284.0677.
4.3.7. Methyl 3-deoxy-3-(4-(4-methoxyophenyl)-1H-[1,2,3]-
triazol-1-yl)-1-thio-b-D-galactopyranoside 23
Compound 9 gave 23 (C18 RP-HPLC eluent H2O–acetonitrile
gradient, 5.4 mg, 72%). 1H NMR (400 MHz, CD3OD) d 8.30 (s, 1H,
H-50), 7.76–7.72 (m, 2H, Ph), 7.00–6.97 (m, 2H, Ph), 4.79 (dd, 1H,
H-3), 4.48 (d, 1H, J1,2 = 9.4, H-1), 4.26 (dd, 1H, J2,3 = 10.6, H-2),
4.13 (d, 1H, J3,4 = 2.9, H-4), 3.82 (s, 3H, CH3O), 3.80–3.68 (m, 3H,
H-5, 2H-6), 2.27 (s, 3H, CH3S). 13C NMR (100.6 MHz, CD3OD) d
161.3 (C-4-Ph), 148.3, 124.5 (C-40, C-1-Ph), 128.0, 115.3 (each 2C,
Ph), 121.0 (C-50), 88.7 (C-1), 81.1 (C-5), 69.8 (C-4), 69.1 (C-3),
67.8 (C-2), 62.4 (C-6), 55.8 (CH3O), 12.1 (CH3S). FAB-HRMS calcd
for C16H21N3NaO5S [M+Na]+ 390.1100; found 390.1103.
4.3.2. Methyl 3-deoxy-3-(4-propyl-1H-[1,2,3]-triazol-1-yl)-1-
thio-b-D-galactopyranoside 18
Compound 4 gave 18 (column eluent CH2Cl2–MeOH 20:1,
1
5.5 mg, 77%). H NMR (400 MHz, CD3OD) d 7.82 (s, 1H, H-50),
4.72 (dd, 1H, J3,4 = 3.0, H-3), 4.44 (d, 1H, J1,2 = 9.4, H-1), 4.17 (dd,
1H, J2,3 = 10.4, H-2), 4.07 (d, 1H, H-4), 3.78–3.67 (m, 3H, H-5,
2H-6), 2.67 (t, 2H, JH,H = 7.5, CH2), 2.25 (s, 3H, CH3S), 1.69 (m, 2H,
CH2), 0.97 (t, 3H, JH,H = 7.3, CH3). 13C NMR (100.6 MHz, CD3OD) d
148.4 (C-40), 122.8 (C-50), 88.8 (C-1), 81.1 (C-5), 69.8 (C-4), 68.8
(C-3), 67.7 (C-2), 62.4 (C-6), 28.5 (CH2), 23.8 (CH2), 14.1 (CH3),
12.1 (CH3S). FAB-HRMS calcd for C12H22N3O4S [M+H]+ 304.1331;
found, 304.1346.
4.3.8. Methyl 3-deoxy-3-(4-(2-fluorophenyl)-1H-[1,2,3]-triazol-
1-yl)-1-thio-b-D-galactopyranoside 24
Compound 10 gave 24 (C18 RP-HPLC eluent H2O–acetonitrile
gradient, 6.0 mg, 82%). 1H NMR (400 MHz, CD3OD) d 8.37 (d, 1H,
JH,F = 3.5, H-50), 8.11 (dd, 1H, J = 1.7, 7.7, Ar), 7.38–7.36 (m, 1H,
Ar), 7.28 (dt, 1H, J = 1.1, 7.7, Ar), 7.22 (ddd, 1H, J = 0.9, 8.2, 9.2,
Ar), 4.86 (obscured under HDO, H-3), 4.49 (d, 1H, J1,2 = 9.3, H-1),
4.86 (t, 1H, J2,3 = 10.3, H-2), 4.15 (d, 1H, J3,4 = 2.8, H-4), 3.83–3.68
(m, 3H, H-5, 2H-6), 2.27 (s, 3H, CH3S). 13C NMR (100.6 MHz,
CD3OD) d 160.6 (d, JC,F = 248, C-2-Ph), 141.7 (d, JC,F = 2.4, C-40),
130.8 (d, JC,F = 8.6, Ar), 128.6 (d, JC,F = 3.3, Ar), 125.8 (d, JC,F = 3.6,
Ar), 124.3 (d, JC,F = 12.1, C-50), 119.7 (d, JC,F = 12.1, C-1-Ph), 116.9
(d, JC,F = 21.7, Ar), 88.7 (C-1), 81.0 (C-5), 69.8 (C-4), 69.1 (C-3),
67.7 (C-2), 62.4 (C-6), 12.0 (CH3S). FAB-HRMS calcd for
4.3.3. Methyl 3-deoxy-3-(4-hydroxymethyl-1H-[1,2,3]-triazol-1-
yl)-1-thio-b-D-galactopyranoside 19
Compound 5 gave 19 (column eluent CH2Cl2–MeOH 12:1, 5.6 mg,
80%). 1H NMR (400 MHz, CD3OD) d 8.01 (s, 1H, H-50), 4.76 (dd, 1H,
J2,3 = 10.6, H-3), 4.69 (s, 2H, CH2), 4.46 (d, 1H, J1,2 = 9.3, H-1), 4.19
(dd, 1H, H-2), 4.09 (d, 1H, J3,4 = 2.8, H-4), 3.79–3.65 (m, 3H, H-5,
2H-6), 2.25 (s, 3H, CH3). 13C NMR (100.6 MHz, CD3OD) d 148.4
(C-40), 123.8 (C-50), 88.7 (C-1), 81.0 (C-5), 69.8 (C-4), 68.9 (C-3),
67.7 (C-2), 62.4 (C-6), 56.6 (CH2), 12.1 (CH3S). FAB-HRMS
C10H17N3NaO5S [M+Na]+ 314.0787; found, 314.0791.
C
15H18FN3NaO4S [M+Na]+ 378.0900; found 378.0902.
4.3.4. Methyl 3-deoxy-3-(4-(1-hydroxycyclohexyl)-1H-[1,2,3]-
triazol-1-yl)-1-thio-b-
D
-galactopyranoside 20
4.3.9. Methyl 3-deoxy-3-(4-(1-naphthyl)-1H-[1,2,3]-triazol-1-
yl)-1-thio-b- -galactopyranoside 25
Compound 6 gave 20 (column eluent CH2Cl2–MeOH 17:1,
D
1
6.9 mg, 87%). H NMR (300 MHz, CD3OD) d 7.93 (s, 1H, H-50), 4.74
Compound 11 gave 25 (C18 RP-HPLC eluent H2O–acetonitrile
(dd, 1H, J2,3 = 10.6, J3,4 = 3.0, H-3), 4.45 (d, 1H, J1,2 = 9.4, H-1), 4.18
gradient, 6.5 mg, 87%). 1H NMR (300 MHz, CD3OD) d 8.38 (s, 1H,

B. A. Salameh et al. / Bioorg. Med. Chem. 18 (2010) 5367–5378
5375
H-50), 8.31–8.27 (m, 1H, Ar), 7.95–7.89 (m, 2H, Ar), 7.70 (dd, 1H,
JH,H = 1.1, 7.1, Ar), 7.57–7.49 (m, 3H, Ar), 4.92 (dd, 1H, H-3 partially
obscured under HDO peak), 4.53 (d, 1H, J1,2 = 9.4, H-1), 4.32 (dd,
1H, J2,3 = 10.5, H-2), 4.22 (d, 1H, J3,4 = 2.6, H-4), 3.87–3.70 (m, 3H,
H-5, 2H-6), 2.28 (s, 3H, CH3S). 13C NMR (75 MHz, CD3OD) d
147.1, 135.4, 132.6, 130.1, 129.5, 129.3, 128.4, 127.7, 127.2,
126.4, 126.3 (C-40, naphthyl), 124.7 (C-50), 88.8 (C-1), 81.1 (C-5),
69.9 (C-4), 69.2 (C-3), 67.8 (C-2), 62.4 (C-6), 12.1 (CH3S). FAB-
HRMS C19H21N3NaO4S [M+Na]+ 410.1150; found, 410.1150.
(C-1), 79.6 (C-5), 68.2 (C-4), 67.5 (C-3), 66.4 (C-2), 61.1 (C-6),
53.0 (CH3O), 11.8 (CH3S). FAB-HRMS calcd for C11H17N3NaO6S
[M+Na]+ 342.0734; found, 342.0723.
4.4. General procedure for the preparation of the amides 29–34
and 39–40
The methyl ester 3 (10 mg) for 30–34, the diester 15 (10 mg) for
29, or the methyl ester 37 (10 mg) for 38 and 39 were stirred with
solution of amine (x) in methanol or water for (t) time and at (T)
temperature. The residue obtained after the evaporation of the sol-
vent was purified by column chromatography using the eluent
indicated.
4.3.10. Methyl 3-deoxy-3-(4-(3-pyridyl)-1H-[1,2,3]-triazol-1-yl)-
1-thio-b-D-galactopyranoside 26
Compound 12 gave 26 (C18 RP-HPLC eluent H2O–acetonitrile
gradient, 5.9 mg, 81%). 1H NMR (400 MHz, DMSO-d6) d 9.40–8.90
(br s, 2H, Ar), 8.72 (s, 1H, H-50), 8.24 (d, 1H, JH,H = 7.9, Ar), 7.57
(br s, 1H, Ar), 5.46 (d, 1H, JH,OH = 7.1, OH-2), 5.27 (d, 1H, JH,OH = 6.5,
OH-4), 4.81 (dd, 1H, J2,3 = 10.6, J3,4 = 2.8, H-3), 4.70 (t, 1H,
JH,OH = 5.8, OH-6), 4.46 (d, 1H, J1,2 = 9.2, H-1), 4.11 (m, 1H, H-2),
3.92 (dd, 1H, H-4), 3.70 (t, 1H, J5,6 = 6.4, H-5), 3.56–3.47 (m, 2H,
2H-6), 2.15 (s, 3H, CH3S). 13C NMR (100.6 MHz, DMSO-d6) d
146.3, 142.9, 132.0, 124.3 (C-40, Ar), 121.8 (C-50), 86.3 (C-1), 79.3
(C-5), 67.6 (C-4), 67.2 (C-3), 66.0 (C-2), 60.3 (C-6), 11.3 (CH3S).
FAB-HRMS calcd for C14H18N4NaO4S [M+Na]+ 361.0946; found
361.0949.
4.4.1. Methyl 3-deoxy-3-(4,5-bis-methylaminocarbonyl-1H-
[1,2,3]-triazol-1-yl)-1-thio-b-D-galactopyranoside 29
Diester 15, x = methylamine (40% in H2O, 2 mL), t = 12 h, col-
umn eluent CH2Cl2–MeOH 10:1 gave 29 (6.8 mg, 92%). 1H NMR
(400 MHz, D2O) d 5.38 (dd, 1H, J2,3 = 10.2, H-3), 4.72 (t, 1H, H-2),
4.64 (d, 1H, JH,H = 7.6, H-1), 4.28 (d, 1H, H-4), 3.96 (dd, 1H, H-5),
3.80 (dd, 1H, J6a,6b = 11.7, J5,6a = 7.2, H-6a), 3.74 (dd, 1H, J5,6a = 5.0,
H-6b), 2.96, 2.95 (2s, each 3H, 2CH3N), 2.29 (s, 3H, CH3S). 13C
NMR (100.6 MHz, D2O) d 162.6, 159.5 (2C@O), 139.8, 132.9 (C-40,
C-50), 87.1 (C-1), 79.7 (C-5), 68.5 (C-4), 67.4 (C-3), 65.8 (C-2),
61.2 (C-6), 26.4, 26.2 (2CH3N), 11.7 (CH3S). FAB-HRMS calcd for
4.3.11. Methyl 3-deoxy-3-(4-p-tolylsulfonyl-1H-[1,2,3]-triazol-1-
C
13H22N5O6S1 [M+H]+ 376.1211; found 376.1289.
yl)-1-thio-b-D-galactopyranoside 27
Compound 13 gave 27 (column eluent CH2Cl2–MeOH 17:1,
4.4.2. Methyl 3-(4-methylaminocarbonyl-1H-[1,2,3]-triazol-1-
yl)-3-deoxy-1-thio-b- -galactopyranoside 30
1
5.8 mg, 75%). H NMR (300 MHz, CD3OD) d 8.69 (s, 1H, H-50),
D
7.91 (d, 2H, JH,H = 8.3, o-Ph), 7.41 (d, 2H, JH,H = 8.0, m-Ph), 4.88
(dd, 1H, H-3 partially obscured under HDO peak), 4.43 (d, 1H,
J1,2 = 9.3, H-1), 4.19 (dd, 1H, J2,3 = 10.4, H-2), 4.05 (d, 1H, J3,4 = 2.9,
H-4), 3.78–3.60 (m, 3H, H-5, 2H-6), 2.46, 2.25 (2s, each 3H,
Methyl ester 3, x = methylamine (40% in H2O, 2 mL), t = 12 h,
T = rt, and column eluent CH2Cl2–MeOH 15:1 gave 30 (7.0 mg,
1
98%). H NMR (400 MHz, CD3OD) d 8.43 (s, 1H, H-50), 4.84 (dd,
1H, J2,3 = 10.6, J3,4 = 3.0, H-3), 4.46 (d, 1H, J1,2 = 9.2, H-1), 4.19 (dd,
1H, H-2), 4.10 (d, 1H, H-4), 3.80–3.67 (m, 3H, H-5, 2H-6), 2.92 (s,
3H, CH3N), 2.25 (s, 3H, CH3S). 13C NMR (100.6 MHz, CD3OD) d
163.4 (C@O), 143.5 (C-40), 126.6 (C-50), 88.6 (C-1), 81.0 (C-5),
69.6 (C-4), 69.1 (C-3), 67.7 (C-2), 62.4 (C-6), 26.1 (CH3N), 12.0
(CH3S). FAB-HRMS calcd for C11H18N4NaO5S [M+Na]+ 341.0896;
found, 341.0892.
13
2CH3). C NMR (75 MHz, CD3OD) d 149.6, 146.6, 139.1 (C-40,
C-1-Ph, C-4-Ph), 131.1, 129.0 (each 2C, Ph), 128.0 (C-50), 88.5
(C-1), 80.8 (C-5), 69.6 (C-3), 69.4 (C-4), 67.5 (C-2), 62.3 (C-6),
21.6 (CH3Ph), 11.9 (CH3S). FAB-HRMS calcd for C16H21N3NaO6S2
[M+Na]+ 438.0770; found 438.0782.
4.3.12. Methyl 3-deoxy-3-(5-p-tolylsulfonyl-1H-[1,2,3]-triazol-1-
yl)-1-thio-b-
D
-galactopyranoside 28
4.4.3. Methyl 3-(4-butylaminocarbonyl-1H-[1,2,3]-triazol-1-yl)-
Compound 14 gave 28 (C18 RP-HPLC eluent H2O–acetonitrile
gradient, 6.2 mg, 81%). 1H NMR (300 MHz, CD3OD) d 8.24 (s, 1H,
H-40), 7.92 (d, 2H, o-Ph), 7.47 (d, 2H, m-Ph), 4.97 (dd, 1H,
J3,4 = 2.8, H-3), 4.66 (t, 1H, JH,H = 9.8, H-2), 4.42 (d, 1H, J1,2 = 9.6,
H-1), 3.75–3.67 (m, 2H, H-5, 2H-6), 3.57 (dd, 1H, J6a,6b = 13.5,
J5,6b = 8.0, H-6b), 2.65, 2.25 (2s, each 3H, 2CH3). 13C NMR
(75 MHz, CD3OD) d 147.9, 139.9, 138.2 (C-50, C-1-Ph, C-4-Ph),
138.1 (C-50), 131.7, 129.2 (each 2C, Ph), 88.7 (C-1), 80.9 (C-5),
69.2, 69.1 (C-3, C-4), 66.8 (C-2), 62.2 (C-6), 21.7 (CH3Ph), 11.8
(CH3S). FAB-HRMS calcd for C16H21N3NaO6S2 [M+Na]+ 438.0770;
found 438.0770.
3-deoxy-1-thio-b-D-galactopyranoside 31
Methyl ester 3, x = butylamine (20% in MeOH, 1 mL), t = 12 h,
T = rt, and column eluent CH2Cl2–MeOH 25:1 gave 31 (7.2 mg,
1
90%). H NMR (400 MHz, D2O) d 8.54 (s, 1H, H-50), 5.01 (dd, 1H,
J2,3 = 10.7, H-3), 4.65 (d, 1H, J1,2 = 9.6, H-1), 4.32 (t, 1H, H-2), 4.22
(d, 1H, J3,4 = 2.8, H-4), 3.98 (dd, 1H, H-5), 3.80 (dd, 1H,
J6a,6b = 11.7, J5,6a = 7.3, H-6a), 3.73 (dd, 1H, J5,6b = 5.0, H-6b), 3.40
(t, 2H, JH,H = 7.0, CH2N), 2.28 (s, 3H, CH3S), 1.59 (m, 2H, CH2), 1.37
(m, 2H, CH2), 0.91 (t, 3H, JH,H = 7.0, CH3). 13C NMR (100.6 MHz,
D2O) d 162.3 (C@O), 142.5 (C-40), 126.4 (C-50), 87.0 (C-1), 79.7
(C-5), 68.3 (C-4), 67.4 (C-3), 66.4 (C-2), 61.1 (C-6), 39.5 (CH2N),
30.9 (CH2), 19.8 (CH2), 13.3 (CH3CH2), 11.8 (CH3S). FAB-HRMS calcd
for C14H25N4O5S [M+H]+ 361.1546; found, 361.1542.
4.3.13. Methyl 3-(4-methoxycarbonyl-1H-[1,2,3]-triazol-1-yl)-3-
deoxy-1-thio-b-D-galactopyranoside 17
Compound 3 (10 mg, 0.023 mmol) was dissolved in methanol
(1.5 mL) and stirred over night at room temperature with sodium
methoxide solution 1 M (0.5 mL). The mixture was neutralized
with Duolite C 436 (H+) resin, filtered and concentrated in vacuo.
The residue was purified by column chromatography (CH2Cl2–
MeOH 25:1) to give 17 (5 mg, 70%). 1H NMR (400 MHz, D2O) d
8.73 (s, 1H, H-50), 5.03 (dd, 1H, J3,4 = 3.0, J2,3 = 10.7, H-3), 4.65 (d,
1H, J1,2 = 9.6, H-1), 4.31 (t, 1H, H-2), 4.22 (d, 1H, H-4), 3.98 (dd,
1H, H-5), 3.95 (s, 3H, CH3O), 3.80 (dd, 1H, J6a,6b = 11.8, J5,6a = 7.3,
H-6a), 3.73 (dd, 1H, J5,6b = 5.0, H-6b), 2.28 (s, 3H, CH3S). 13C NMR
(100.6 MHz, D2O) d 162.8 (C@O), 139.5 (C-40), 129.0 (C-50), 87.0
4.4.4. Methyl 3-(4-butylaminocarbonyl-1H-[1,2,3]-triazol-1-yl)-
3-deoxy-1-thio-b-D-galactopyranoside 31
Methyl ester 3, x = butylamine (20% in MeOH, 1 mL), t = 12 h,
T = rt, and column eluent CH2Cl2–MeOH 25:1 gave 31 (7.2 mg,
1
90%). H NMR (400 MHz, D2O) d 8.54 (s, 1H, H-50), 5.01 (dd, 1H,
J2,3 = 10.7, H-3), 4.65 (d, 1H, J1,2 = 9.6, H-1), 4.32 (t, 1H, H-2), 4.22
(d, 1H, J3,4 = 2.8, H-4), 3.98 (dd, 1H, H-5), 3.80 (dd, 1H,
J6a,6b = 11.7, J5,6a = 7.3, H-6a), 3.73 (dd, 1H, J5,6b = 5.0, H-6b), 3.40
(t, 2H, JH,H = 7.0, CH2N), 2.28 (s, 3H, CH3S), 1.59 (m, 2H, CH2), 1.37
(m, 2H, CH2), 0.91 (t, 3H, JH,H = 7.0, CH3). 13C NMR (100.6 MHz,
D2O) d 162.3 (C@O), 142.5 (C-40), 126.4 (C-50), 87.0 (C-1), 79.7 (C-

5376
B. A. Salameh et al. / Bioorg. Med. Chem. 18 (2010) 5367–5378
5), 68.3 (C-4), 67.4 (C-3), 66.4 (C-2), 61.1 (C-6), 39.5 (CH2N), 30.9
(CH2), 19.8 (CH2), 13.3 (CH3CH2), 11.8 (CH3S). FAB-HRMS calcd
for C14H25N4O5S [M+H]+ 361.1546; found, 361.1542.
4.4.9. Methyl 3-(4-benzylaminocarbonyl-1H-[1,2,3]-triazol-1-
yl)-3-deoxy-b- -galactopyranosyl-(1–4)-2-acetamido-2-deoxy-
b- -glucopyranoside 40
Methyl ester 37, x = benzylamine (20% in MeOH, 2 mL), t = three
D
D
4.4.5. Methyl 3-(4-(3-hydroxyprop-1-yl-aminocarbonyl)-1H-
days, T = 40 °C, RP-HPLC (C18, H2O–acetonitrile gradient) gave 40
1
[1,2,3]-triazol-1-yl)-3-deoxy-1-thio-b-
D-galactopyranoside 32
(6.6 mg, 81%). H NMR (400 MHz, CD3OD) d 8.45 (s, 1H, H-500),
Methyl ester 3, x = 3-aminopropanol (20% in MeOH, 1 mL),
t = two days, T = 45 °C, column eluent CH2Cl2–MeOH 10:1 gave
7.35–7.23 (m, 5H, Ph), 4.87 (dd partially obscured under HDO peak,
1H, H-30), 4.64 (d, 1H, J1 ,2 = 7.6, H-10), 4.58 (s, 2H, CH2), 4.33 (d, 1H,
0
0
1
32 (7.0 mg, 86%). H NMR (400 MHz, D2O) d 8.56 (s, 1H, H-50),
J1,2 = 8.2, H-1), 4.14 (dd, 1H, J2 ,3 = 11.1, H-20), 4.02 (d, 1H, J3 ,4 = 2.8,
H-40), 3.91–3.63 (m, 8H, H-2, H-3, H-4, 2H-6, H-50, 2H-60), 3.46 (s,
3H, CH3O), 3.39 (m, 1H, H-5), 1.97 (s, 3H, CH3C@O). 13C NMR
(100.6 MHz, CD3OD) d 173.6, 162.7 (2C@O), 173.5, 140.0 (C-40,
C-1-Ph), 129.5, 128.5, 128.3 (Ph), 126.9 (C-50), 105.32 (C-10),
103.6 (C-1), 80.6, 77.8, 76.6, 74.3, 69.4, 69.3, 67.6, 62.6, 61.7,
57.0, 56.6 (C-5, C-50, C-4, C-40, C-3, C-30, C-2, C-20, C-6, C-60,
CH3O), 43.7 (CH2), 22.9 (CH3S). FAB-HRMS C25H35N5NaO11
[M+Na]+ 604.2231; found, 604.2230.
0
0
0
0
5.01 (dd, 1H, J3,4 = 3.0, J2,3 = 10.6, H-3), 4.65 (d, 1H, J1,2 = 9.6, H-1),
4.32 (t, 1H, H-2), 4.22 (d, 1H, H-4), 3.98 (dd, 1H, H-5), 3.80 (dd,
1H, J5,6a = 7.4, J6a,6b = 11.8, H-6a), 3.73 (dd, 1H, J5,6b = 5.0, H-6b),
3.69 (t, 2H, JH,H = 6.4, CH2O), 3.49 (t, 2H, JH,H = 6.9, CH2N), 2.28 (s,
3H, CH3S), 1.87 (m, 2H, CH2). 13C NMR (100.6 MHz, D2O) d 162.4
(C@O), 142.5 (C-40), 126.4 (C-50), 87.0 (C-1), 79.7 (C-5), 68.3
(C-4), 67.4 (C-3), 66.4 (C-2), 61.1 (C-6), 59.6 (CH2O), 36.7 (CH2N),
31.3 (CH2), 11.8 (CH3S). FAB-HRMS calcd for C13H22N4NaO6S
[M+Na]+ 385.1158; found, 385.1180.
4.4.10. Methyl 3-(4-aminocarbonyl-5-amino-1H-[1,2,3]-triazol-
1-yl)-3-deoxy-1-thio-b-D-galactopyranoside 35
4.4.6. Methyl 3-(4-(N0-morpholino)ethylaminocarbonyl)-1H-
[1,2,3]-triazol-1-yl)-3-deoxy-1-thio-b-
D
-galactopyranoside 33
Cyanoacetamide (7.0 mg, 3 equiv) and K2CO3 (11.5 mg, 3 equiv)
in DMSO (0.5 mL) were stirred for 1 h. After this time a solution of
the azide 1 (10 mg, 0.028 mmol) in DMSO was added. The resulting
mixture was stirred at 50 °C overnight. The solvent was removed
and the residue was kept in methanol overnight. After evaporation
of the methanol, the residue was purified by column chromatogra-
phy CH2Cl2–MeOH 20:1 to give 35 (5.2 mg, 60%). 1H NMR
(400 MHz, CD3OD) d 4.54 (dd, 1H, H-2), 4.46 (dd, 1H, J3,4 = 2.7,
J2,3 = 10.3, H-3), 4.41 (d, 1H, J1,2 = 9.2, H-1), 4.19 (d, 1H, H-4),
3.79–3.68 (m, 3H, H-5, 2H-6), 2.24 (s, 3H, CH3). 13C NMR
(100.6 MHz, CD3OD) d 167.2 (C = O), 147.5, 123.1 (C-40, C-50), 88.9
(C-1), 81.0 (C-5), 69.5 (C-4), 66.4 (C-3), 65.9 (C-2), 62.4 (C-6),
11.9 (CH3S). FAB-HRMS calcd for C10H17N5NaO5S [M+Na]+
342.0848; found 342.0852.
Methyl ester 3, x = N-morpholinoethylamine (20% in MeOH,
1 mL), t = four days, T = 45 °C, column eluent CH2Cl2–MeOH 17:1
gave 33 (7.0 mg, 75%). 1H NMR (400 MHz, CD3OD) d 8.44 (s, 1H,
H-50), 4.85 (obscured under HDO peak, 1H, H-3), 4.46 (d, 1H,
J1,2 = 9.2, H-1), 4.19 (dd, 1H, J2,3 = 10.4, H-2), 4.09 (d, 1H, J3,4 = 2.9,
H-4), 3.80–3.66 (m, 7H, H-5, 2H-6,
2 (CH2O)), 3.55 (t, 2H,
JH,H = 6.6, CH2NC@O), 2.59 (t, 2H, JH,H = 6.6, CH2N), 2.53 (br t, 4H,
2 (CH2N), 2.25 (s, 3H, CH3S). 13C NMR (100.6 MHz, CD3OD) d
162.7 (C@O), 143.5 (C-40), 126.7 (C-50), 88.6 (C-1), 80.9 (C-5),
69.6 (C-4), 69.1 (C-3), 67.8 (–CH2OCH2–), 67.7 (C-2), 62.3 (C-6),
58.5 (CH2N), 54.7 ((CH2)2N), 36.9 (CH2NC@O), 12.0 (CH3S). FAB-
HRMS calcd for
440.1579.
C
16H27N5NaO6S [M+Na]+ 440.1580; found
4.4.7. Methyl 3-(4-benzylaminocarbonyl-1H-[1,2,3]-triazol-1-
yl)-3-deoxy-1-thio-b- -galactopyranoside 34
4.4.11. Methyl 3-deoxy-3-(4-(1-naphthyl)-1H-[1,2,3]-triazol-1-
D
yl)-b-D-galactopyranosyl-(1–4)-2-acetamido-2-deoxy-b-D-
Methyl ester 3, x = benzylamine (20% in MeOH, 1 mL), t = three
days, T = 45 °C, column eluent CH2Cl2–MeOH 25:1 gave 34 (7.0 mg,
glucopyranoside 41
Compound 38 (10 mg, 0.013 mmol) was stirred in a solution of
methylamine in water 40% (2 mL) for 12 h, concentrated, and puri-
fied by RP-HPLC (C18, H2O–acetonitrile gradient) to give 41
1
80%). H NMR (400 MHz, D2O) d 8.58 (s, 1H, H-50), 7.41–7.35 (m,
5H, Ph), 5.02 (dd, 1H, J2,3 = 10.7, H-3), 4.65 (d, 1H, J1,2 = 9.6, H-1),
4.62 (s, 2H, CH2), 4.32 (t, 1H, H-2), 4.23 (d, 1H, J3,4 = 2.8, H-4),
3.98 (dd, 1H, H-5), 3.80 (dd, 1H, J5,6a = 7.4, J6a,6b = 11.7, H-6a),
3.73 (dd, 1H, J5,6b = 5.0, H-6b), 2.28 (s, 3H, CH3S). 13C NMR
(100.6 MHz, D2O) d 162.4 (C@O), 142.4 (C-40), 138.1, 129.2 [2C],
127.9, 127.6 [2C] (Ph), 126.6 (C-50), 87.0 (C-1), 79.7 (C-5), 68.3
(C-4), 67.4 (C-3), 66.4 (C-2), 61.1 (C-6), 43.2 (CH2Ph), 11.8 (CH3S).
FAB-HRMS calcd for C17H22N4NaO5S [M+Na]+ 417.1209; found,
417.1224.
1
(5.5 mg, 75%). H NMR (400 MHz, CD3OD) d 8.34 (s, 1H, H-500),
8.27 (m, 1H, Ar), 7.93 (m, 2H, Ar), 7.70 (dd, 1H, JH,H = 1.1, 7.1, Ar),
7.53 (m, 3H, Ar), 4.93 (dd, 1H, J2 ,3 = 11.1, J3 ,4 = 3.0, H-30), 4.70 (d,
0
0
0
0
1H, J1 ,2 = 7.5, H-10), 4.34 (d, 1H, J1,2 = 8.3, H-1), 4.28 (dd, 1H,
H-20), 4.15 (d, 1H, H-40), 3.97–3.63 (m, 8H, H-3, H-4, H-50, 2H-60,
2H-6, H-2), 3.46 (s, 3H, CH3O), 3.42 (m, 1H, H-5), 1.97 (s, 3H,
CH3C@O). 13C NMR (100.6 MHz, CD3OD) d 173.6 (C@O), 147.1,
135.4, 132.6, 130.1, 129.5, 129.3, 128.4, 127.7, 127.2, 126.4, 126.3
(C-400, naphthalene), 124.7 (C-500), 105.4 (C-10), 103.6 (C-1), 80.8,
77.9, 76.7, 74.4, 69.7, 69.3, 67.7, 62.3, 61.8, 57.0, 56.7 (C-2, C-3,
C-4, C-5, C-6, C-20, C-30, C-40, C-50, C-60, CH3O), 22.9 (CH3C@O).
FAB-HRMS C27H34N4NaO10S [M+Na]+ 597.2173; found, 597.2181.
0
0
4.4.8. Methyl 3-(4-methylaminocarbonyl-1H-[1,2,3]-triazol-1-
yl)-3-deoxy-b-
b- -glucopyranoside 39
Methyl ester 37, x = methylamine (40% in H2O, 2 mL), t = 12 h,
T = rt, column eluent CH2Cl2–MeOH 5:1 gave 39 (5.6 mg, 80%). 1H
D-galactopyranosyl-(1–4)-2-acetamido-2-deoxy-
D
4.4.12. 1,2,4,6-Tetra-O-acetyl-3-deoxy-3-(4-(methoxycarbonyl)-
NMR (400 MHz, D2O) d 8.55 (s, 1H, H-500), 5.01 (dd, 1H, J3 ,4 = 2.9,
1H-1,2,3-triazol-1-yl)-b-
A mixture of 1,2,4,6-tetra-O-acetyl-4-azido-4-deoxy-b-
topyranose 4234 (66 mg, 76
mol), methyl propiolate (16.4
1 equiv), CuI (3.5 mg, 0.1 equiv), diisopropylethylamine (32
1 equiv) and toluene (6 mL) was stirred overnight, concentrated
and purified by flash chromatography (2:1, heptane–ethyl acetate)
to give 43 (75.0 mg, 93%). 1H NMR (400 MHz, CDCl3) 2.21, 2.15,
2.09, 2.08, 1.88, 1.87 (3H each, s, 8CH3C@O), 2.04 (6H, s, 2CH3C@O),
3.95 (6H, s, OCH3), 4.07–4.21 (4H, m, 2H-6, 2H-60), 4.24 (1H, at,
D-galactopyranose 43
0
0
J2 ,3 = 11.2, H-30), 4.74 (d, 1H, J1 ,2 = 7.6, H-10), 4.67 (d, 1H,
J1,2 = 7.8, H-1), 4.24 (dd, 1H, H-20), 4.17 (d, 1H, H-40), 4.00 (m, 2H,
H-50, H-6a), 3.87–3.72 (m, 6H, H-2, 2H-60, H-3, H-4, H-6b), 3.61
(m, 1H, H-5), 3.50 (s, 3H, CH3O), 2.94 (s, 3H, CH3N), 2.04 (s, 3H,
CH3C@O). 13C NMR (100.6 MHz, D2O) d 175.1, 162.9 (2C@O),
142.4 (C-400), 126.3 (C-500), 103.4 (C-10), 102.2 (C-1), 76.4 (C-50),
75.1 (C-5), 68.2 (C-20), 68.0 (C-40), 66.0 (C-30), 60.3 (C-6), 57.5
(CH3O), 55.4 (C-60), 78.8, 72.9, 61.1 (C-2, C-3, C-4), 26.0 (CH3N),
22.5 (CH3C@O). FAB-HRMS calcd for C19H32N5O11 [M+H]+
506.2099; found, 506.2101.
D
-galac-
0
0
0
0
l
l
l
L,
L,
J = 6.5, H-5b), 4.56 (1H, at, J = 6.5, H-5
J2,3 = 11.0 Hz, J3,4 = 3,3, H-3b), 5.40 (1H, dd, J2,3 = 11.8, J3,4 = 3,3 Hz,
a), 5.22 (1H, dd,

B. A. Salameh et al. / Bioorg. Med. Chem. 18 (2010) 5367–5378
5377
H-3
a
), 5.54 (1H, d, H-4b), 5.59 (1H, d, H-4
a
), 5.83 (2H, m, H-1b and
), 8.16 and
3.52–3.55 (4H, m, H-6, H-60), 3.70 (2H, m, H-5), 3.91 (2H, m,
H-4), 4.10 (2H, m, H-2), 4.70 (2H, at, JOH,6 = 5.7, OH-6), 4.87–4.93
(4H, m, H-1, H-3), 5.26 (2H, d, JOH,4 = 7.5, OH-4), 5.49 (2H, d,
JOH,2 = 6.8, OH-2), 8.36 (2H, s, triazole-H), 8.44 (1H, q, NH). FAB-
HRMS calcd for C20H31N8O10S [M+H]+ 575.1884; Found 575.1887.
H-2b), 5.92 (1H, dd, J1,2 = 3.5, H-2
a), 6.50 (1H, d, H-1
a
8.10 (1H, s, triazole-H). 13C NMR (100.6 MHz, CDCl3) 20.0, 20.1,
20.2 (2C), 20.5 (2C), 20.6, 20.7 (8CH3C@O), 52.3 (OCH3), 58.2,
60.8, 61.0, 62.2. 65.2, 66.5, 67.9, 68.1, 68.9, 72.7, 88.9, 92.4 (C-1,
C-2, C-3, C-4, C-5, C-6), 126.6 (triazole-CH), 140.1, 140.2 (tria-
zole-C), 160.5, 160.6, 168.5, 168.6, 168.7, 168.9, 169.0, 169.3,
170.1, 170.2 (10CH3C@O). FAB-HRMS calcd for C18H23N3NaO11
[M+Na]+ 480.1230; Found 480.1234.
4.5. General procedure for deprotection and formation of
amides 47–52
The methyl ester 45 (4–8 mg) was stirred with solution of
amine (x) 20% in methanol (1 mL) for (t) time. The mixture was
concentrated in vacuo and the residue was purified by column
chromatography to give the deprotected amides 47–52. In the
cases of the less volatile amines, the crude material was left on
the high vacuum pump for several days before chromatography.
4.4.13. 2,4,6-Tri-O-acetyl-3-deoxy-3-(4-(methoxycarbonyl)-1H-
1,2,3-triazol-1-yl)-b-
Compound 43 (28 mg, 61
ane (1 ml) that had been dried over 4 Å molecular sieves. Acetic
anhydride (12 l, 0.12 mmol) and HBr (0.2 ml of a 33% solution
D-galactopyranosyl bromide 44
lmol) was dissolved in dichlorometh-
l
in AcOH) were added, and the mixture was stirred under N2 at
room temperature. After 2 h 30 min, the reaction mixture was di-
luted with dichloromethane (30 ml) and washed with NaHCO3
(30 ml of a saturated aqueous solution), dried (MgSO4), filtered
and concentrated in vacuo. The residue was purified by flash col-
umn chromatography (1:1, heptane–ethyl acetate, 1% Et3N) to give
4.5.1. Di-(3-deoxy-3-(4-((butylamino)carbonyl)-1H-1,2,3-
triazol-1-yl)-b-D-galactopyranosyl)sulfane 47
x = butylamine, t = 26 h, column eluent CH2Cl2–MeOH 9:1 gave
47 (quant.) as a white solid. 1H NMR (400 MHz, CD3OD) 0.96 (6H, t,
J = 7.4, NHCH2CH2CH2CH3), 1.42 (4H, m, NHCH2CH2CH2), 1.60 (4H,
m, NHCH2CH2), 3.39 (4H, t, J = 7.1, NHCH2), 3.67 (2H, dd, J5,6 = 4.0,
44 (24 mg, 82%) as a colourless oil; ½a D21
ꢂ
+177 (c 1.0 in CHCl3). 1H
J6,6 = 11.1, H-6), 3.77–3.86 (4H, m, H-5, H-60), 4.10 (2H, d, J3,4 = 2.8,
0
NMR (300 MHz, CDCl3) d 1.91 (3H, s, CH3C@O), 2.02 (6H, s,
0
2CH3C@O), 3.91 (3H, s, OCH3), 4.09 (1H, dd, J5,6 = 6.7, J6,6 = 11.5,
H-4), 4.75 (2H, at, J = 9.9, H-2), 4.82 (2H, d, J1,2 = 9.5, H-1), 4.90 (2H,
dd, J2,3 = 10.4, H-3), 8.55 (2H, s, triazole-H). 13C NMR (100.6 MHz,
CD3OD) 14.1 (q, NHCH2CH2CH2CH3), 21.1 (t, NHCH2CH2CH2), 32.7
(t, NHCH2CH2), 39.9 (t, NHCH2), 62.8 (t, C-6), 68.3 (d, C-2), 68.7
(d, C-3), 69.5 (d, C-4), 81.4 (d, C-5), 86.7 (d, C-1), 126.8 (d, tria-
zole-CH), 143.6 (s, triazole-C), 162.7 (s, C@O). FAB-HRMS calcd
for C26H42N8NaO10S [M+Na]+ 681.2642; Found 681.2648.
H-6), 4.19 (1H, dd, J5,6 = 6.2, H-60), 4.61 (1H, at, J = 6.4, H-5), 5.39
0
(1H, dd, J2,3 = 11.4, J3,4 = 1.6, H-3), 5.59 (1H, d, H-4), 5.72 (1H, dd,
J1,2 = 3.8, H-2), 6.84 (1H, d, H-1), 8.82 (1H, s, triazole-H). 13C NMR
(75 MHz, CDCl3) 20.4, 20.5, 20.7 (3q, 3CH3C@O), 52.4 (q, OCH3),
59.1, 60.9, 66.9, 67.7, 71.2, 88.3 (5 ꢃ d, t, C-1, C-2, C-3, C-4, C-5,
C-6), 128.0 (d, triazole-CH), 140.1 (s, triazole-C), 160.8 (s, CH3C@O),
169.2, 169.5, 170.3 (3s, 3CH3C@O). FAB-HRMS calcd for C16H20
-
N3O9BrNa [M+Na]+ 500.0281; Found 500.0291.
4.5.2. Di-(3-deoxy-3-(4-((benzylamino)carbonyl)-1H-1,2,3-
triazol-1-yl)-b-D-galactopyranosyl)sulfane 48
4.4.14. Di-(2,4,6-tri-O-acetyl-3-deoxy-3-(4-(methoxycarbonyl)-
1H-1,2,3-triazol-1-yl)-b-D-galactopyranosyl) sulfane 45
x = benzylamine, t = 26 h, column eluent CH2Cl2–MeOH 9:1
gave 48 (quant.) as a white solid. 1H NMR (400 MHz, CD3OD)
0
Sodium sulfate nonahydrate (170 mg, 0.59 mmol) was dried by
dissolving in water and then heating to 50 °C under high vacuum.
Molecular sieves 4 Å (ca. 100 mg) were added. Compound 44
(77 mg, 0.16 mmol) was dissolved in distilled acetonitrile (5 ml)
and added to the reaction vessel. The mixture was stirred at room
temperature for 13 h. After this time, TLC (1:3, heptane–ethyl ace-
tate) indicated the complete consumption of starting material (Rf
0.7) and the presence of a major product (Rf 0.2). The reaction mix-
ture was poured onto a plug of silica and washed through with
ethyl acetate–chloroform, 1:1. The eluent was concentrated in va-
cuo and the residue was purified by flash column chromatography
(1:4, heptane–ethyl acetate) to give 45 (24 mg, 36%). White crys-
3.67 (2H, dd, J5,6 = 4.0, J6,6 = 11.1, H-6), 3.77–3.85 (4H, m, H-5,
H-60), 4.09 (2H, d, J3,4 = 2.8, H-4), 4.58 (4H, ABq, JAB = 12.0, NHCH2),
4.77 (2H, at, J = 9.9, H-2), 4.82 (2H, d, J1,2 = 9.3, H-1), 4.90 (2H, dd,
J2,3 = 10.0, H-3), 7.22–7.36 (10H, m, Ar-H), 8.60 (2H, s, triazole-H).
13C NMR (100.6 MHz, CD3OD) 43.8 (t, NHCH2), 62.8 (t, C-6), 68.3
(d, C-2), 68.7 (d, C-3), 69.5 (d, C-4), 81.4 (d, C-5), 86.8 (d, C-1),
127.1, 128.2, 128.5, 129.5 (4 ꢃ d, Ar-CH, triazole-CH), 140.0,
143.5 (2 ꢃ s, Ar-C, triazole-C), 162.8 (s, C@O). FAB-HRMS calcd
for C32H38N8NaO10S [M+Na]+ 749.2329; Found 749.2333.
4.5.3. Di-(3-deoxy-3-(4-((2-phenylethylamino)carbonyl)-1H-
1,2,3-triazol-1-yl)-b-D-galactopyranosyl)sulfane 49
tals, mp (MeOH) >220 °C; ½a D21
ꢂ
+30.2 (c 0.17 in MeOH–CHCl3,
x = phenethylamine, t = 28 h, column eluent CH2Cl2–MeOH 9:1
gave 49 (quant.)as a white solid. 1H NMR (400 MHz, DMSO-d6)
2.84 (4H, at, J = 7.5, NHCH2), 3.47–3.56 (8H, m, NHCH2CH2, H-6,
H-60), 3.71 (2H, at, J = 6.2, H-5), 3.92 (2H, m, H-4), 4.14 (2H, m,
H-2), 4.71 (2H, at, JOH,6 = 5.7, OH-6), 4.87–4.93 (4H, m, H-1, H-3),
5.27 (2H, d, JOH,4 = 7.4, OH-4), 5.49 (2H, d, JOH,2 = 6.9, OH-2), 7.18–
7.31 (10H, m, Ar-H), 8.37 (2H, s, triazole-H) 8.54 (1H, t, J = 5.9,
NH). FAB-HRMS calcd for C34H42N8NaO10S [M+Na]+ 777.2642;
Found 777.2651.
1:1). 1H NMR (400 MHz, CDCl3) d 1.92, 2.07, 2.10 (18H, 3s,
6CH3C@O), 3.94 (6H, s, OCH3), 4.10–4.24 (6H, m, H-5, H-6, H-60),
5.03 (1H, d, J1,2 = 9.7, H-1), 5.25 (1H, dd, J2,3 = 11.0, J3,4 = 3.2, H3),
5.60 (1H, d, H-4), 5.72 (1H, at, J = 10.3, H-2), 8.18 (1H, s, triazole-
H). 13C NMR (100.6 MHz, CDCl3) 20.5, 20.6, 20.8 (3q, 3CH3C@O),
52.3 (q, OCH3), 61.4 (t, C-6), 63.4 (d, C-3), 66.5 (d, C-2), 68.5 (d,
C-4), 75.8 (d, C-5), 82.4 (d, C-1), 126.8 (d, triazole-CH), 140.4 (s, tri-
azole-C), 160.7 (s, CH3C@O), 169.0, 169.5, 170.5 (3 s, 3CH3C@O).
FAB-HRMS calcd for C32H40N6NaO18S [M+Na]+ 851.2017; Found
851.2017.
4.5.4. Di-(3-deoxy-3-(4-((allylamino)carbonyl)-1H-1,2,3-triazol-
1-yl)-b-D-galactopyranosyl)sulfane 50
4.4.15. Di-(3-deoxy-3-(4-((methylamino)carbonyl)-1H-1,2,3-
x = allylamine, t = 28 h, column eluent CH2Cl2–MeOH 9:1 gave
triazol-1-yl)-b-
D
-galactopyranosyl)sulfane 46
mol) was suspended in a methyl-
50 (quant.) as a white solid. 1H NMR (400 MHz, CD3OD) 3.67 (2H,
dd, J5,6 3.8, J6,6 = 10.8, H-6), 3.76–3.86 (4H, m, H-5, H-60), 4.01
0
Compound 45 (6.5 mg, 7.8
l
amine solution (40% in water, 2 ml). The mixture was stirred at
room temperature for 48 h. After this time, the mixture was
concentrated in vacuo to give 46 (4.5 mg, quant.) as a white solid.
1H NMR (400 MHz, DMSO-d6) d 2.77 (3H, d, J = 4.7, NHCH3),
(4H, d, J = 5.3, NHCH2), 4.10 (2H, d, J3,4 = 2.6, H-4), 4.75 (2H, at,
J = 9.8, H-2), 4.81 (2H, d, J1,2 = 9.4, H-1), 4.91 (2H, dd (obs), H-3),
5.13 (1H, dd, J = 1.4, JZ = 10.3, CH@CHEHZ), 5.22 (1H, dd, J = 1.5,
JE = 17.2, CH@CHEHZ), 5.92 (2H, m, CH@CH2), 8.57 (2H, s,

5378
B. A. Salameh et al. / Bioorg. Med. Chem. 18 (2010) 5367–5378
triazole-H). 13C NMR (100.6 MHz, CD3OD) 42.4 (t, NHCH2), 62.8 (t,
C-6), 68.3 (d, C-2), 68.7 (d, C-3), 69.5 (d, C-4), 81.4 (d, C-5), 86.7 (d,
C-1), 116.3 (t, CH@CH2), 127.0 (d, triazole-CH), 135.4 (d, CH@CH2),
143.5 (s, triazole-C), 162.7 (s, C@O). FAB-HRMS calcd for
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C
24H34N8NaO10S [M+Na]+ 649.2016; Found 649.2010.
4.5.5. Di-(3-deoxy-3-(4-((2-hydroxypropylamino)carbonyl)-1H-
1,2,3-triazol-1-yl)-b- -galactopyranosyl)sulfane 51
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D
x = 2-hydroxypropylamine, t = 28 h, column eluent CH2Cl2–
MeOH 9:2 gave 51 (70%) as a white solid. 1H NMR (400 MHz,
DMSO-d6) 1.66 (2H, quint., J = 6.5, NHCH2CH2), 3.3 (4H, m (obs),
NHCH2), 3.45 (4H, m, NHCH2CH2CH2), 3.51–3.56 (4H, m, H-6,
H-60), 3.71 (2H, at, J = 6.1, H-5), 3.93 (2H, m, H-4), 4.15 (2H, m,
H-2), 4.50 (2H, t, J = 5.2, CH2CH2OH), 4.71 (2H, at, JOH,6 = 5.7,
OH-6), 4.87–4.93 (4H, m, H-1, H-3), 5.27 (2H, d, JOH,4 = 7.3, OH-4),
5.48 (2H, d, JOH,2 = 6.8, OH-2), 8.37 (2H, s, triazole-H), 8.48 (1H, t,
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J = 5.8, NH). FAB-HRMS calcd for C24H38N8NaO12
685.2228; Found 685.2231.
S
[M+Na]+
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4.5.6. Di-(3-deoxy-3-(4-((2-methoxyethylamino)carbonyl)-1H-
1,2,3-triazol-1-yl)-b- -galactopyranosyl)sulfane 52
D
x = 2-methoxyethylamine, t = 28 h, column eluent CH2Cl2–
MeOH 9:1 gave 52 (quant.) as a white solid. 1H NMR (400 MHz,
DMSO-d6) 3.26 (6H, s, OCH3), 3.41–3.46 (8H, m, NHCH2, H-6,
H-60), 3.50–3.55 (4H, m, NHCH2CH2), 3.70 (2H, at, J = 6.4, H-5),
3.93 (2H, m, H-4), 4.12 (2H, m, H-2), 4.71 (2H, at, JOH,6 = 5.8,
OH-6), 4.88–4.93 (4H, m, H-1, H-3), 5.28 (2H, d, JOH,4 = 7.4, OH-4),
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685.2214.
Acknowledgement
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This work was supported by the Swedish Research Council
(Grant No. 621-2003-4265), the programs ‘Glycoconjugates in Bio-
logical Systems’ and ‘Chemistry for Life Sciences’ sponsored by the
Swedish Strategic Research Foundation, the foundation ‘Olle Engk-
vist Byggmästare’, and the Royal Pysiographic Society, Lund.
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Supplementary data
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Supplementary data associated with this article can be found, in
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