Synthesis of multivalent glycoclusters from 1-thio-β-D-galactose and their inhibitory activity against the β-galactosidase from E. coli

Article abstract of DOI:10.1021/jo102421e

The synthesis of multivalent glycoclusters, designed to be compatible with biological systems, is reported. A variety of 1-thio-β-d-galactosides linked to a terminal triple bond through oligoethyleneglycol chains of variable lengths has been synthesized. Also, azide-containing oligosaccharide scaffolds were prepared from trehalose, maltose, and maltotriose by direct azidation with NaN₃/PPh₃/CBr₄. Click reaction between the thiogalactoside residues and the azide scaffolds under microwave irradiation afforded a family of glycoclusters containing 1 to 4 residues of 1-thio-β-D-galactose. The yields went from moderate to excellent, depending on the valency of the desired product. Deacetylation with Et ₃N/MeOH/H₂O led to the final products. Complete characterization of the products was performed by NMR spectroscopy and HR-MS techniques. Their activities as inhibitors of β-galactosidase from E. coli were determined by using the Lineweaver-Burk method. The use of hydrophilic carbohydrate scaffolds for the synthesis of multivalent galactosides represents an interesting approach to improve their pharmacokinetics and bioavailability. In addition, the presence of the thioglycosidic bond will improve their stability in biological fluids.

Full text of DOI:10.1021/jo102421e

ARTICLE
pubs.acs.org/joc
Synthesis of Multivalent Glycoclusters from 1-Thio-β-D-galactose and
Their Inhibitory Activity against the β-Galactosidase from E. coli
Alejandro J. Cagnoni,† Oscar Varela,† Sꢀebastien G. Gouin,‡ Josꢀe Kovensky,*,‡ and María Laura Uhrig*,†
†CIHIDECAR-CONICET, Departamento de Química Orgꢀanica, Facultad de Ciencias Exactas y Naturales,
Universidad de Buenos Aires, Buenos Aires, Argentina
‡LABORATOIRE DES GLUCIDES-CNRS, Universitꢀe de Picardie Jules Verne, Amiens, France
S Supporting Information
b
ABSTRACT: The synthesis of multivalent glycoclusters, de-
signed to be compatible with biological systems, is reported. A
variety of 1-thio-β-D-galactosides linked to a terminal triple
bond through oligoethyleneglycol chains of variable lengths has
been synthesized. Also, azide-containing oligosaccharide scaf-
folds were prepared from trehalose, maltose, and maltotriose by
direct azidation with NaN3/PPh3/CBr4. Click reaction between the thiogalactoside residues and the azide scaffolds under
microwave irradiation afforded a family of glycoclusters containing 1 to 4 residues of 1-thio-β-D-galactose. The yields went from
moderate to excellent, depending on the valency of the desired product. Deacetylation with Et3N/MeOH/H2O led to the final
products. Complete characterization of the products was performed by NMR spectroscopy and HR-MS techniques. Their activities
as inhibitors of β-galactosidase from E. coli were determined by using the LineweaverꢀBurk method. The use of hydrophilic
carbohydrate scaffolds for the synthesis of multivalent galactosides represents an interesting approach to improve their
pharmacokinetics and bioavailability. In addition, the presence of the thioglycosidic bond will improve their stability in biological
fluids.
’ INTRODUCTION
are operative.9 The interaction of lactose-functionalized gold
glyconanoparticles with a lectin and with the β-galactosidase
from Escherichia coli has been evaluated.10 The proper selection
of ligand densities and spacers led to an increased resistance of
the lactose moiety of the nanoparticle to hydrolysis by the
enzyme.
Carbohydrateꢀprotein interactions are involved in cellular
recognition processes that include viral and bacterial infections,
inflammation, and tumor metastasis.1 Recognition events, occur-
ring in the active sites of glycosyltransferases and glycosyl
hydrolases, account for the metabolism of carbohydrates. In
some natural systems, the sometimes weak binding affinity of
carbohydrates to their receptors is overcome by a multivalent
display of sugar residues at the surface of cells, which leads to the
so-called “glycoside clustering effect”.
During the last years, multivalent ligands having β-galactoside
or β-lactoside epitopes attached to a variety of scaffolds have
been synthesized.2 Most of them present O-linked saccharides
and their preparation involves classic glycosylating methods. To
connect the epitope to the scaffold, oligoethyleneglycol (EG)
linkers are usually employed, due to their wide biomedical
applications,3 flexibility, and ability to prevent nonspecific ad-
sorption to proteins.4 The preparation of a variety of multivalent
ligands differing in EG linker lengths can contribute to elucidate
the mechanisms involved in the recognition processes.5,6 The
“cluster effect” has been extensively studied for several lectin
systems,7 but little is known about the influence of the multi-
valency on the activity of glycosidases. So far, we have shown that
a trivalent iminosugar displayed a 6-fold affinity enhancement
toward Jack bean R-mannosidase.8 Also, a recent report on the
multivalent effect of fullerene iminosugar balls on the inhibition
of R-glucosidases suggests that alternative binding mechanisms
The β-galactosidase from E. coli is highly specific for
β-galactopyranosyl nonreducing moieties, which may be linked
to a variety of aglycons.11 The enzyme is able to recognize C- and
S-galactosyl residues that, in turn, may act as moderate to good
inhibitors of its hydrolytic activity.12 The interaction of this
β-galactosidase with several substrates has been studied by
X-ray13 and NMR12,14,15 experiments, and we have synthesized
a family of thiodisaccharides that display an important inhibitory
activity against the enzyme.16,17 In contrast to natural O-linked
sugars, the S-carbohydrate mimetics are usually resistant to
metabolic processes18,19 and are seen as potential precursors of
promising carbohydrate-based therapeutics.20 Particularly, the
resistance of glycoclusters to enzyme hydrolysis is crucial to
circumvent degradation when they are internalized in the cell.
When designing glycoclusters to study biological processes,
specificity, affinity, and stability against glycolytic enzymes are
important factors to be considered. We assumed that the
replacement of the interglycosidic oxygen atom by a sulfur atom
Received: December 7, 2010
Published: March 29, 2011
r
2011 American Chemical Society
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Scheme 1. Synthesis of Alkynyl-Armed S-Galactosides 3aꢀc
Scheme 2. Azide-Containing Oligosaccharide Scaffolds 4ꢀ10
in glycoclusters could increase the resistance to hydrolytic degrada-
tion by enzymes and can even cause inhibition of this activity.
Therefore, we report here the synthesis of a family of glycoclusters
based on carbohydrate scaffolds bearing one to four S-galactoside
epitopes and EG spacers of different lengths. The copper-catalyzed
azideꢀalkyne cycloaddition (CuAAC)21 reaction was employed as
the key step to connect the recognition elements to the scaffolds.
The inhibition behavior against E. coli β-galactosidase was studied
in order to determine the existence of cluster effects.
a triplet, and signals at ∼80 and ∼74 ppm for the linked and
terminal alkyne carbon atoms, respectively). The resonances of the
nonequivalent protons of the methylene groups vicinal to the sulfur
appeared well resolved in a clean region of the spectrum (2.85ꢀ
3.58 ppm).
A variety of azide-containing scaffolds were prepared accord-
ing to previously reported methods (Scheme 2). Thus, com-
pound 4 was prepared by treatment of cyclopentanol with CBr4/
PPh3/NaN3. Reaction of glucose pentaacetate and azidotri-
methylsilane in the presence of SnCl4 afforded β-1-azidotetra-
acetilglucopyranose (5) in 94% yield.23 Sugar scaffolds 6ꢀ10 were
prepared by the direct azidation method reported by Kovensky,
starting from the corresponding free sugar, sodium azide, PPh3,
and CBr4 in anhydrous DMF, followed by acetylation.24
This methodology was also employed for the preparation of
the 6,60-diazido derivative 8 from trehalose, which was obtained
in an excellent yield (80%), in comparison with previously
reported methods using trehalose/PPh3/CBr4/LiN3 (13%)25
or by regioselective tosylation of C-6 and C-60 followed by
substitution with NaN3 (14%).26
In the first instance, the coupling of thioglycoside 3a with
cyclopentylazide (4) was conducted as a model reaction, in order
to verify if the conditions of the click reaction were compatible
with the thioglycosides (Scheme 3). The reaction between 3a
and 4, in the presence of copper sulfate and sodium ascorbate in
DMF/H2O, was completed at room temperature within 10ꢀ16 h.
’ RESULTS AND DISCUSSION
The syntheses of the S-thiogalactosides (3aꢀc) having a
terminal triple bond linked through spacers of different lengths
were achieved by treatment of the thiouronium salt derived from
galactose (1) with the appropriate bromides 2aꢀc (Scheme 1).
Linkers 2b and 2c were prepared by standard methods, starting
from diethyleneglycol or pentaethyleneglycol and propargylbro-
mide, followed by bromination. Thus, reaction of 1 with bro-
mides 2a, 2b, or2c, in the presence of NEt3,22 led to thiogalactosides
3aꢀc, respectively, in excellent isolated yields (71ꢀ100%). The
spectra of compounds 3aꢀc showed the characteristic signals of the
H-1 protons in the region of 4.51ꢀ4.75 ppm, protected by the
proximity of the sulfur atom, in accordance with those reported for
other 1-thiogalactopyranosides.17 Also, the diagnostic signals for the
1
alkyne group in both the H and 13C NMR spectra were clearly
observed (2.27ꢀ2.48 ppm for the alkyne proton, which appeared as
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Scheme 3. Synthesis of Monovalent Thiogalactosides 11ꢀ16, Using the Click Reaction As the Key Step
However, the reaction conducted under microwave irradiation
required a much shorter reaction time (40ꢀ50 min), as reported
for related systems.27 These conditions proved to be very efficient in
the case of the multivalent structures (polyazides) described below.
In all cases, the consumption of the starting materials was followed
by TLC and MS spectrometry. The latter technique was particularly
useful to analyze reaction mixtures that involved polar partners, such
as 3b and 3c, difficult to detect by TLC.
smaller values would be expected for 1,5-disubstituted
regioisomers.28 The methylene protons of the linker next to the
sulfur appeared as two multiplets in the region 4.14ꢀ2.17 ppm,
whereas the methylene protons of the ethyleneglycol moiety
(CH2O) showed a complex signal at ∼3.60 ppm. The resonances
of H-6 and H-60 in compounds 15aꢀc were observed downfield,
with respect to those of 13aꢀc, due to the deshielding effect of the
triazole ring at C-6. The collection of all these data served as a base
for the spectroscopic analysis of the products of superior valency
described below.
The click reaction between alkynes 3aꢀc and the monoazides
5 or 6, conducted under the conditions optimized for the
coupling of 3a and 4, afforded the monovalent compounds
Two families of divalent cycloadducts 17aꢀc and 19aꢀc were
prepared (Scheme 4), adjusting the reaction conditions to
guarantee the reaction of the two azide groups, present in the
scaffolds [a 2:1 molar ratio a alkyne (3aꢀc):diazide (7 or 8) was
employed]. The reactions proceeded under microwave irradia-
tion to afford the click products in high yields. Thus, the scaffold
7, derived from glucose, led to adducts 17aꢀc in 84ꢀ60% yield.
The NMR spectra of these compounds were rather complex, in
particular those of 17a, due to the proximity of the signals of the
two nonequivalent thiogalactose residues. Their structures were
confirmed by comparison with the spectroscopic data of the
monovalent compounds described above and with the assistance
1
13aꢀc and 15aꢀc in 72ꢀ100% isolated yields. The H NMR
spectrum of these monovalent products showed the signals of
both sugar residues, the glucose-scaffold and the thiogalactose,
and a first order analysis was admitted. The assignments were
further confirmed by 2D NMR techniques. In the description of
the spectra, the signals corresponding to the thiogalactose moiety
were labeled as “G”. The thiogalactose anomeric protons (H-1G)
of compounds 13aꢀc and 15aꢀc appeared at 4.33ꢀ4.63 ppm,
whereas those of the Glcp moiety in 13aꢀc, bonded to the
nitrogen of triazole, appeared at a lower field (5.83ꢀ5.90 ppm).
The aromatic proton of the triazole ring was identified as a singlet
at δ 7.5ꢀ8.4. Accordingly, the 13C NMR spectrum showed the
characteristic signals at approximately 144 and 123 ppm respectively
13
of 2D 1H COSY and 1Hꢀ C HSQC experiments.
For compound 17a, the diagnostic resonance of H-1 (Glc
residue) was observed as a doublet at 5.89 ppm, with a coupling
constant of 8.9 Hz (β-configuration), and the resonances of the
β-thiogalactoses anomeric protons, namely H-1G and H-1G0,
for C-4 and C-5 of the triazole ring. In all cases, the large Δ(δC-4
δC-5) values observed for the different triazoles, ranging from 19 to
ꢀ
25 ppm, corroborated the 1,4-disubstituted pattern since much
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Scheme 4. Synthesis of Multivalent Thiogalactosides 17ꢀ24
appeared at 4.58 and 4.29 ppm (J1,2 ≈ 10.0 Hz). As the length of
the spacer increased in the sequence 17a f 17b f 17c, the
signals of both thiogalactose residues became more similar, and
in fact, they appeared as identical in 17c.
Reaction of the diazide scaffold 8 derived from trehalose with
thiogalactosides 3aꢀc afforded the symmetric divalent ligands
19aꢀc in good yields (∼70%). One major advantage of the use
of scaffold 8, in terms of the characterization of the products, was
the simplicity of their NMR spectra, similar to those of thiodi-
saccharides,16,17,19b,29 as a result of the symmetry. Comparison of
1H and 13C NMR spectra showed a conserved resonance pattern
in the series 19a f 19b f 19c not only for the “trehalose core”,
but also for the thiogalactose residue. Molecular weights, deter-
mined by MS, confirmed the dimeric structures.
To synthesize trivalent ligands, the maltose-derived triazide
scaffold 9 was prepared. Cycloaddition reaction between 9 and
thioglycosides 3aꢀc led to compounds 21aꢀc in good yields
(54ꢀ67%). The trivalent glycoclusters 21b and 21c appeared to
be quite polar, due to the EG-spacer, and mixtures of EtOAc/MeOH
were required for their purification by column chromatography. The
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NMR spectra, although highly complex were clear enough to
confirm their structures. In agreement with the formation of the
three triazole rings were observed three aromatic protons signals
at 7.81ꢀ8.20 ppm and three pairs of aromatic carbons at ∼145
and ∼125 ppm. The two anomeric protons of maltose appeared
0
0
at ∼6.2 (J1,2 ≈ 9.0 Hz) and 5.46 ppm (J1 ,2 = 3.6 Hz),
corresponding to the β- and R-glucose residues (H-1 and H-10,
respectively). These signals correlated with peaks at 84.3 and
96.6 ppm, respectively, in the 13C NMR spectrum. The reso-
nances for the anomeric carbons of the three thiogalactose
residues were detected at 82.8, 82.7, and 82.4 ppm for 21a, but
appeared as a single peak (83.6 ppm) for 21b and 21c. Further-
more, MS confirmed the structures proposed.
Tetravalent ligands 23aꢀc were synthesized (∼50% yield)
from the tetraazide-scaffold 10, derived from maltotriose, and
thioglycosides 3aꢀc. Diagnostic signals were observed in the 1H
NMR spectrum: four aromatic proton signals (8.2ꢀ7.7 ppm)
and four pairs of aromatic carbon signals (∼145 and ∼125 ppm).
The MS confirmed the structures.
Figure 1. LineweaverꢀBurk plot for inhibition of E. coli β-glactosidase
by compound 24a at different concentrations: ([) 0.00, (9 ) 0.40, (2)
0.80, and (b) 1.20 mM of inhibitor.
All the glycocluster derivatives obtained were O-deacetylated
by treatment with Et3N/MeOH/H2O 1:4:5 (Schemes 3 and 4),
and the reactions were followed by TLC. After desalting with an
anion exchange resin and purification by reverse-phase chroma-
tography, the deprotected products were recovered in good
yields, and analyzed by NMR recorded for D2O solutions. As
the complexity of the spectra increased with the number of
galactose residues attached to the platforms, complete and
detailed assignment of the signals was not possible for all the
products. Signals of anomeric protons and carbons, correspond-
ing both to the sugar scaffold and the thiogalactose residues, were
diagnostic and are shown as a table in the Supporting Informa-
tion. MS and elemental analysis of the products confirmed the
structures of the free glycoclusters.
Table 1. Ki Values for the Inhibition of β-Galactosidase from
E. coli by the β-S-Galactoside Glycoclusters, Determined by
Using o-Nitrophenyl-β-D-galactopyranoside As Substrate, As
Described in the Experimental Sectiona
Ki value (mM)
valency
compd
a
b
c
1
1
1
2
2
3
4
12
14
16
18
20
22
24
0.901 ( 0.009
1.506 ( 0.009
0.283 ( 0.003
0.507 ( 0.007
0.419 ( 0.004
0.438 ( 0.006
0.254 ( 0.003
1.192 ( 0.009
1.008 ( 0.007
0.713 ( 0.008
0.487 ( 0.005
0.386 ( 0.004
0.262 ( 0.003
1.063 ( 0.009
1.102 ( 0.008
0.711 ( 0.006
0.490 ( 0.004
0.382 ( 0.004
0.255 ( 0.003
a The inhibition was competitive in all cases. (Km = 1.378 ( 0.006 mM).
’ EVALUATION OF THE INHIBITORY ACTIVITY
The resistance of the new products to enzymatic hydrolysis
was evaluated by using the β-galactosidase from E. coli. The
glycoclusters were dissolved in sodium phosphate buffer (pH
7.3) in concentrations identical with that of the o-nitrophenyl
β-D-galactopyranoside substrate employed for the determination
of the Km. The solutions were incubated with the enzyme at
37 °C. To determine the stability of the glycoclusters toward the
enzyme, aliquots of the solution were taken and examined by
TLC and NMR. Even compound 20b, which is a weak inhibitor
of the enzyme as discussed later, was recovered practically intact
after 24 h of incubation. Therefore, the glycoclusters seem to be
slow substrates, or no substrates at all, of the enzyme and
resistant to enzymatic hydrolysis.
The glycoclusters act as inhibitors of the β-galactosidase from
E. coli as their presence in the incubation medium inhibited the
hydrolysis of the o-nitrophenyl β-D-galactopyranoside employed
as substrate of the enzyme. The kinetics of the inhibition was
determined by the double reciprocal plot method, as exemplified
in Figure 1 for compound 24a.
enhancing the flexibility and facilitating the accommodation of
the thiogalactoside residue in the active site.
The comparison of Ki values for the monovalent derivatives
shows the influence of the scaffold on the β-galactosidase
inhibition as well. When the β-thiogalactose epitope is attached
through a triazole ring to the C-6 of the scaffold (as in compound
16a) an increment in the inhibition is observed, compared to the
analogues that have thiogalactose attached to the triazole bonded
to the anomeric position of the scaffold, as in 14a. The increased
inhibitory activity may be attributed to the higher flexibility of the
ligand when linked to C-6, and to the presence of the vicinal
heteroaromatic ring. In agreement with the higher inhibitory
activity shown for compound 16a, we have described that a
β-thiogalactosyl residue attached to a flexible pentopyranose ring
with an anomeric aromatic group results in a strong inhibition of
E. coli β-galactosidase.12,17 Thus, an increase of the linker length
that separates the thiogalactose residue and the triazol, as in
16a f 16b f 16c, results in a decrease of the inhibitory activity
(Ki from 0.283 f 1.008 f 1.102). Therefore, when comparing
the monovalent series 14aꢀc and 16aꢀc, the length of the linker
has an opposite effect, which seems to depend on the relative
position of the triazole on the scaffold.
All the glycoclusters are competitive inhibitors of the enzyme
in the micromolar range (Table 1). The inhibition constants (Ki)
are similar to those determined for other β-thiogalactosides
previously reported.30
As the scaffold for compounds 18aꢀc is a 1,6-bistriazole-
substituted monosaccharide, we can speculate that combined
effects of the same kind as those operating in compounds 14aꢀc
The length of the linker influences the inhibitory activity of the
molecule. For the series 14a f 14b f 14c, it seems that longer
linkers contribute to an increase of the inhibition, probably by
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and 16aꢀc are involved. Thus, the Ki values for the series 18a f
18b f 18c show that an increase in the distance between the
thiosugar and the triazole ring at C-6 in the scaffold diminishes
the inhibitory activity. This effect is opposite, and seems to be
stronger, than the favorable effect imparted by longer linkers
between the thiogalactose and the triazole bonded to C-1. The
same tendency is observed for derivatives 20a f 20b f 20c,
which exhibited a somewhat higher activity, probably due to the
role of the scaffold in the global shape of the glycocluster, that
may affect the ability of the epitopes to reach the active site.
The Ki values listed in Table 1 show that moving from
monovalent (12, 14aꢀc, 16bꢀc) to di- (18aꢀc, 20aꢀc), tri-
(22aꢀc), and tetravalent (24aꢀc) derivatives, increased inhibi-
tion is observed. However, this is probably due to a statistical
effect, and no cluster or multivalency effect is observed for
β-galactosidase inhibition. Cluster effects of multivalent azasu-
gars on enzyme inhibition have been previously observed,8 but
only for some enzymes (for example, R-mannosidase), whereas
for others no such effect was detected. Nevertheless, our present
results show that the multivalent thiogalactosides are stronger
inhibitors of galactosidase than the monomeric counterparts
(with the exception of compound 16a).
bromide (0.60 mL, 7.08 mmol) was added dropwise and the resulting
mixture was stirred for 6 h at rt. Methanol (5 mL) was added. The mixture
was concentrated under reduced pressure and the residue was dissolved in
EtOAc (20 mL). The organic layer was washed with saturated ammonium
chloride (20 mL), the aqueous layer was extracted with EtOAc (2 ꢁ
20 mL), and the combined organic extracts were concentrated. The residue
was purified by flash chromatography to give the corresponding alkyny-
lalcohols (60ꢀ66% yield) as colorless oils, which were employed for the
bromination step. Thus, the alkynylalcohols (0.634 mmol) and CBr4 (232
mg, 0.697 mmol) were dissolved in CH2Cl2 (1 mL) and PPh3 (183 mg,
0.697 mmol) was added slowly while the mixture was stirred at 0 °C. The
resulting mixture was stirred at rt until TLC and MS indicated the
disappearance of the starting materials (4 h). The mixture was concentrated
under reduced pressure and the residue was subjected to flash chromatog-
raphy with hexane/EtOAc (1:1) as eluent.
Compound 2b: yield 94%; Rf 0.49 (hexane/EtOAc 1:1); 1H NMR
(500 MHz, CDCl3) δ 4.22 (d, 2 H, J1,3 = 2.4 Hz, H-3), 3.82 (t, 2 H, J8,9
=
6.3 Hz, H-8), 3.71 (s, 4 H, H-5, H-6), 3.48 (t, 2 H, J8,9 = 6.3 Hz, H-9),
2.44 (t, J1,3 = 2.4 Hz, H-1); 13C NMR (125 MHz, CDCl3) δ 79.5 (C-2),
74.6 (C-1), 71.2 (C-8), 70.3, 69.0 (C-5, C-6), 58.5 (C-3), 30.2 (C-9).
Anal. Calcd for C7H11BrO2: C, 40.60; H, 5.35. Found: C, 40.52; H, 5.09.
HRMS (ESI) m/z [M þ Na]þ calcd for C7H11BrNaO2 298.9835, found
298.9844.
Compound 2c: yield 82%; Rf 0.33 (hexane/EtOAc 1:1); 1H NMR
(500 MHz, CDCl3) δ 4.21 (d, 2 H, J1,3 = 2.4 Hz, H-3), 3.81 (t, 2 H, J8,9
6.3 Hz, H-8), 3.69ꢀ3.64 (s, 16 H, H-5, H-6, H-8, H-9, H-11, H-12, H-14,
H-15), 3.48 (t, 2 H, J8,9 = 6.3 Hz, H-9), 2.43 (t, J1,3 = 2.4 Hz, H-1); 13
NMR (125 MHz, CDCl3) δ 79.7 (C-2), 74.5 (C-1), 71.2 (C-17), 70.3,
69.0 (C-5, C-6, C-8, C-9, C-11, C-12, C-14, C-15), 58.4 (C-3), 30.4 (C-
18). Anal. Calcd for C13H23BrO5: C, 46.03; H, 6.83. Found: C, 46.33; H,
6.56. HRMS (ESI) m/z [M þ Na]þ calcd for C13H23BrNaO5 361.0627,
found 361.0615.
=
’ CONCLUSIONS
In conclusion, we report here an efficient and optimized
synthetic strategy to obtain multivalent ligands bearing 1-thio-
β-D-galactose residues, as promising molecules for in vivo studies.
The reactions employed proved to be compatible with the thiogly-
cosidic bonds. The acetylated precursors, as well as the free
glycoclusters, were fully characterized by NMR spectroscopy and
MS spectrometry. Moreover, as far as we know, this is the first report
on the inhibitory behavior of multivalent thioglycoclusters.
These compounds are also interesting as putative inhibitors of
galectin-mediated processes. Multivalent ligands having O-linked
β-galactoside or β-lactoside residues attached to a variety of
scaffolds have been synthesized and evaluated as inhibitors of
lectins.2 Also, some S-galactosides have shown inhibitory activity
toward galectins.31 Studies on the affinity of the glycoclusters
reported here to lectins are in progress.
C
General Procedure for the Synthesis of the Thiogalacto-
sides 3aꢀc. 2,3,4,6-Tetra-O-acetyl-β-D-galactopyranosyl isothiouro-
nium bromide16 (220 mg, 1.063 mmol) and the corresponding
alkylbromide (2a, 2b, or 2c, 1.594 mmol) were dissolved in anhydrous
acetonitrile (4 mL) and NEt3 (371 μL, 2.658 mmol) was added
dropwise. The resulting mixture was stirred at rt for 3 h, when TLC
and MS indicated complete consumption of the starting material. The
solvent was evaporated under reduced pressure and the residue was
purified by flash chromatography (cyclohexane/EtOAc 7:3).
Compound 3a: yield 100%; [R]20D ꢀ47.6 (c 0.7, CHCl3); Rf 0.58
1
(hexane/EtOAc 1:1); H NMR (500 MHz, CDCl3) δ 5.45 (dd, 1 H,
’ EXPERIMENTAL SECTION
J4,5 = 1.0 Hz, J3,4 = 3.4 Hz, H-4), 5.27 (t, 1 H, J1,2 = J2,3 = 10.0 Hz, H-2),
5.10 (dd, 1 H, J3,4 = 3.2 Hz, J2,3 = 10.0 Hz, H-3), 4.75 (d, 1 H, J1,2 = 10.0
Hz, H-1), 4.18 (dd, 1 H, J5,6a = 6.7 Hz, J6a,6b = 11.3 Hz, H-6a), 4.12 (dd, 1
H, J5,6b = 6.5 Hz, J6a,6b = 11.4 Hz, H-6b), 3.97 (ddd, 1 H, J4,5 = 1.1 Hz,
General Methods. Analytical thin layer chromatography (TLC)
was performed on Silica Gel 60 F254 aluminum supported plates (layer
thickness 0.2 mm) with solvent systems given in the text. Visualization of
the spots was effected by exposure to UV light and charring with a
solution of 5% (v/v) sulfuric acid in EtOH, containing 0.5% p-
anisaldehyde. Column chromatography was carried out with Silica Gel
60 (230ꢀ400 mesh). Optical rotations were measured at 20 °C in a 1 cm
cell in the stated solvent; [R]D values are given in 10ꢀ1 deg cm2 gꢀ1
J5,6b = 6.5 Hz, J5,6a = 6.7 Hz, H-5), 3.58, 3.31 (2 dd, 2 H, J = 2.6 Hz, Jgem
=
16.5 Hz, CH2S), 2.27 (t, 1 H, J = J0 = 2.6 Hz, CtCH), 2.08, 2.07, 2.05,
2.04 (4 s, 12 H, 4 CH3CO); 13C NMR (125 MHz, CDCl3) δ 170.6,
169.8, 169.7, 169.6 (ꢀCOCH3), 82.5 (C-1), 78.7 (ꢀCtCH), 74.5
(C-5), 71.9, 71.8 (C-3, CtCH), 67.2 (C-2), 67.0 (C-4), 61.3 (C-6), 20.7,
20.6 (ꢁ2), 20.5 (CH3CO), 17.5 (SCH2). Anal. Calcd for C17H22O9S: C,
50.74; H, 5.51; S, 7.97. Found:C, 50.46; H, 5.77; S, 7.92. HRMS (ESI) m/z
[M þ Na]þ calcd for C17H22NaO9S 425.0882, found 425.0884.
3
3
(concentration c given as g/100 mL). Microwave irradiation was carried
out in a CEM Discover instrument, at 70 °C (power max 300 W). High-
resolution mass spectra HRMS where obtained by Electrospray Ioniza-
tion (ESI) and Q-TOF detection. For 1H, 13C nuclear magnetic
resonance (NMR) spectra, chemical shifts are reported in parts per
million relative to tetramethylsilane or a residual solvent peak (CHCl3:
1H, δ = 7.26; 13C, δ = 77.2). Assignments of 1H and 13C were assisted by
Compound 3b: yield 85%; [R]20 ꢀ6.9 (c 0.8, CHCl3); Rf 0.46
D
1
(hexane/EtOAc 1:1); H NMR (500 MHz, CDCl3) δ 5.43 (dd, 1 H,
J4,5 = 0.9 Hz, J3,4 = 3.4 Hz, H-4), 5.21 (t, 1 H, J1,2 = J2,3 = 10.0 Hz, H-2),
5.05 (dd, 1 H, J3,4 = 3.4 Hz, J2,3 = 10.0 Hz, H-3), 4.65 (d, 1 H, J1,2 = 10.0
Hz, H-1), 4.21 (d, 2 H, J = 2.3 Hz, CH2CtCH), 4.16 (dd, 1 H, J5,6a = 6.6
Hz, J5,6b = 11.3, H-6a), 4.11 (dd, 1 H, J5,6b = 6.6 Hz, J6a,6b = 11.3, H-6b),
3.96 (ddd, 1 H, J4,5 = 0.9 Hz, J5,6a = J5,6b = 6.6 Hz, H-5), 3.77ꢀ3.64 (m, 6
H, CH2O), 3.00 (ddd, 1 H, J = 6.9 Hz, J = 7.0, Jgem = 13.8 Hz, CHS), 2.79
(ddd, 1 H, J = 5.8 Hz, J = 7.0, Jgem = 13.8 Hz, CHS), 2.48 (t, J = 2.3 Hz,
2D 1HꢀCOSY and 2D 1Hꢀ C CORR experiments.
13
General Procedure for the Synthesis of the Alkylbromides
2b and 2c. Diethylene glycol or pentaethylene glycol (4.71 mmol) and
sodium hydride (170 mg, 7.08 mmol) were dissolved in anhydrous DMF
(40 mL) and the mixture was stirred at 0 °C for 30 min. Then, propargyl
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CH2CtCH), 2.16, 2.07, 2.05, 1.98 (4 s, 12 H, 4 CH3CO); 13C NMR
(125 MHz, CDCl3) δ 170.3, 170.2, 170.0, 169.6 (ꢀCOCH3), 84.1 (C-
1), 79.5 (CtCH), 74.7 (C-5), 74.3 (ꢀCtCH), 71.8 (C-3), 71.4, 70.1,
69.0 (CH2O), 67.4, 67.3 (C-4, C-2), 61.4 (C-6), 58.4 (CH2CtCH),
29.4 (CH2S), 20.8, 20.7 ( ꢁ 2), 20.6 (CH3CO). Anal. Calcd for
C21H30O11S: C, 51.42; H, 6.16; S, 6.54. Found: C, 51.40; H, 6.12; S,
6.32. HRMS (ESI) m/z [M þ Na]þ calcd for C21H30NaO11S 513.1407,
found 513.1401.
1 H, J3,4 = 3.4 Hz, J2,3 = 10.0 Hz, H-3), 4.91 (m, 1 H, J = 6.4 Hz, J = 7.5
Hz, CHNcyclo), 4.63 (d, 1 H, J1,2 = 10.0 Hz, H-1), 4.12ꢀ4.08 (m, 3 H,
H-60a, H-60b, CHS), 3.95 (ddd, 1 H, J4,5 = 1.0 Hz, J5 ,6 a = J5 ,6 b = 6.4 Hz,
H-5), 3.93 (d, 1 H, Jgem = 14.3 Hz, CHS), 2.30ꢀ2.23, 2.10ꢀ2.00
1.92ꢀ1.88, 1.79ꢀ1.75 (4 m, 8 H, CH2cyclo), 2.16, 2.05, 2.03, 1.98 (4 s, 12
H, 4 CH3CO); 13C NMR (125 MHz, CDCl3) δ 170.3, 170.2, 170.0,
169.6 (-COCH3), 144.4 (C-4 triazole), 120.4 (C-5 triazole), 83.4 (C-1),
74.4 (C-5), 71.7 (C-3), 67.2 (ꢁ2) (C-2, C-4), 61.8 (C-1cyclo), 61.2 (C-
6), 33.3 (C-2cyclo), 24.5 (CH2S), 24.0 (C-3cyclo), 20.7, 20.6 (ꢁ2), 20.5
(ꢀCOCH3). Anal. Calcd for C22H31N3O9S: C, 51.45; H, 6.08; N, 8.18;
S, 6.24. Found: C, 51.12; H, 6.34; N, 7.98; S, 6.10. HRMS (ESI) m/z
[M þ Na]þ calcd for C22H31N3NaO9S 536.1679, found 536.1697.
Compound 13a: solvent system hexane/EtOAc (1:1); yield 100%;
mp 99ꢀ101 °C; [R]20D ꢀ49.7 (c 0.5, CHCl3); Rf 0.28 (hexane/EtOAc
1:1); 1H NMR (500 MHz, CDCl3) δ 7.85 (s, 1 H, H-triazole), 5.87 (m, 1
H, J1,2 = 9.1 Hz, H-1), 5.47ꢀ5.44 (m, 3 H, H-2, H-3, H-4G), 5.34 (m, 1
H, J3,4 = 9.5 Hz, J4,5 = 10.0 Hz, H-4), 5.24 (t, 1 H, J1G,2G = J2G,3G = 9.9
Hz, H-2G), 5.17 (dd, 1 H, J3G,4G = 3.4 Hz, J2G,3G = 10.0 Hz, H-3G), 4.36
(dd, 1 H, J5G,6aG = 5.2 Hz, J6aG,6bG = 10.5 Hz, H-6aG), 4.33 (d, 1 H,
J1G,2G = 9.9 Hz, H-1G), 4.33 (dd, 1 H, J5,6a = 5.3 Hz, J6a,6b = 12.6 Hz,
H-6a), 4.16 (dd, 1 H, J5,6b = 2.1 Hz, J6a,6b = 12.6 Hz, H-6b), 4.14 (d, 1 H,
Jgem = 14.2 Hz, CHS), 4.09 (ddd, 1 H, J4G,5G = 1.0 Hz, J5G,6aG = 5.1 Hz,
J5G,6bG = 7.3 Hz, H-5G), 4.03 (dd, 1 H, J5G,6bG = 7.5 Hz, J6aG,6bG = 10.5
Hz, H-6bG), 4.02 (ddd, 1 H, J5,6b = 2.1 Hz, J5,6a = 5.2 Hz, J4,5 = 10.0 Hz,
H-5), 3.85 (d, 1 H, Jgem = 14.2 Hz, CHS), 2.19, 2.15, 2.08, 2.07 (ꢁ2),
1.99, 1.97, 1.91 (8 s, 24 H, 8 CH3CO); 13C NMR (125 MHz, CDCl3) δ
170.9, 170.4 (ꢁ2), 170.0, 169.8, 169.6, 169.4 (ꢁ2) (ꢀCOCH3), 143.9
(C-4 triazole), 121.1 (C-5 triazole), 86.0 (C-1), 80.7 (C-1G), 75.2 (C-
5), 73.6 (C-5G), 72.3* (C-2), 71.6 (C-3G), 70.5* (C-3), 67.7 (C-4),
67.5, 67.4 (C-2G, C-4G), 61.6, 61.4 (C-6, C-6G), 23.4 (CH2S), 20.7,
20.6, 20.5, 20.4, 20.1 (CH3CO). Anal. Calcd for C31H41N3O18S: C,
48.00; H, 5.33; N, 5.42; S, 4.13. Found: C, 47.91; H, 5.12; N, 5.12; S,
4.25. HRMS (ESI) m/z [M þ Na]þ calcd for C31H41N3NaO18S
798,2004, found 798,1980.
0
0
0
0
Compound 3c: yield 71%; [R]20D ꢀ12.9 (c 0.7, CHCl3); Rf 0.33
(hexane/EtOAc 1:3); 1H NMR (500 MHz, CDCl3) δ 5.30 (br d, 1 H,
J4,5 < 1.0 Hz, J3,4 = 3.3 Hz, H-4), 5.07 (t, 1 H, J1,2 = J2,3 = 10.0 Hz, H-2),
4.92 (dd, 1 H, J3,4 = 3.4 Hz, J2,3 = 10.0 Hz, H-3), 4.51 (d, 1 H, J1,2 = 10.0
Hz, H-1), 4.07 (d, 2 H, J = 2.4 Hz, CH2CtCH), 4.02 (dd, 1 H, J5,6a = 6.5
Hz, J6a,6b = 11.6 Hz, H-6a), 4.00 (dd, 1 H, J5,6b = 6.7 Hz, J6a,6b = 11.6 Hz,
H-6b), 3.86 (t, 1 H, J5,6a = 6.5 Hz, J5,6b = 6.7 Hz, H-5), 3.60ꢀ3.49 (m, 18
H, 9 CH2O), 2.85, 2.67 (2 m, 2 H, J = 6.8 Hz, Jgem = 13.5 Hz, CH2S),
2.39 (t, 1 H, J = 2.4 Hz, ꢀCtCH), 2.03, 1.94, 1.92, 1.85 (4 s, 12 H, 4
CH3CO); 13C NMR (125 MHz, CDCl3) δ 170.5, 170.4, 170.2, 169.8
(ꢀCOCH3), 84.28 (C-1), 80.1 (ꢀCtCH), 75.0 (ꢀCtCH), 74.7 (C-
5), 72.1 (C-3), 72.1, 71.5, 70.9 (ꢁ4), 70.6 (ꢁ2), 69.4 (CH2O), 67.7
(ꢁ2) (C-2, C-4), 61.8 (C-6), 58.6 (CH2CtCH), 29.7 (CH2S), 21.0,
20.9 (ꢁ2), 20.8 (CH3CO). Anal. Calcd for C27H42O14S: C, 52.08; H,
6.80; S, 5.15. Found: C, 52.05; H, 6.86; S, 5.02. HRMS (ESI) m/z [M þ
Na]þ calcd for C27H42NaO14S 645.2193, found 645.2199.
General Procedures for the Synthesis of Azide-Containing
Scaffolds 5ꢀ10
Procedure A: Compounds 6ꢀ10 were synthesized from the
corresponding free sugar, sodium azide, PPh3, and CBr4 in anhydrous
DMF as previously described.21c,24 These products showed identical
properties to those previously reported.21c,24,26 Synthesis of 8 from
trehalose was achieved in 80% yield, following the same methodology
described to obtain the diazido derivative of maltose (9). Purification
of 8 by column chromatography was performed with hexane/EtOAc
(3:2 f 1:1).
Procedure B: Synthesis of 5: Glucose pentaacetate (150 mg,
0.384 mmol) and azidotrimethylsilane (56 μL, 0.423 mmol) were
dissolved in anhydrous CH2Cl2 (4 mL) and SnCl4 (45 μL, 0.384 mmol)
was added dropwise.23 The resulting mixture was stirred at rt until TLC
indicated the disappearance of starting material (2 h). The mixture was
extracted with NaHCO3 (3 ꢁ 20 mL) and then washed with water
(20 mL). The organic layer was dried (Na2SO4) and filtered, and the
solvent was evaporated under reduced pressure. The residue was
purified by flash chromatography to give 5 (135 mg, 94% yield), mp
128.5, [R]20D ꢀ30.0 (c 0.5, CHCl3) {lit.32 mp 129 °C; [R]D ꢀ33.0 (c
0.7, CHCl3)}.33
General Procedure for the Click Reaction: Synthesis of
Compounds 11, 13aꢀc, 15aꢀc, 19aꢀc, 21aꢀc, and 23aꢀc
The corresponding azido-saccharide 5ꢀ10 (1.00 mmol) and the
selected alkynyl-thiogalactoside 3aꢀc (1.00 mmol per mol of reacting
azide) were dissolved in a dioxane/H2O mixture (8:2 mL, 35 mL).
Copper sulfate (0.25 mmol per mol of reacting azide) and sodium
ascorbate (0.50 mmol per mol of azide reacting group) were added, and
the mixture was stirred at rt until MS indicated the disappearance of
starting materials and intermediates (10ꢀ16 h). Alternatively, the
mixture was stirred under microwave irradiation during 40 min. The
mixture was then poured into a 1:1 H2O/NH4Cl solution (60 mL) and
extracted with EtOAc (4 ꢁ 30 mL). The organic layer was dried
(Na2SO4) and filtered, and the solvent was removed under reduced
pressure. The residue was purified by flash chromatography, using the
solvent systems indicated in each case.
Compound 13b: solvent system hexane/EtOAc (1:1); yield 89%;
[R]20D ꢀ19.7 (c 0.8, CHCl3); Rf 0.34 (hexane/EtOAc 1:3); 1H NMR
(500 MHz, CDCl3) δ 7.83 (s, 1 H, H-triazole), 5.90 (d, 1 H, J1,2 = 9.0
Hz, H-1), 5.46 (dd, 1 H, J1,2 = 9.0 Hz, J2,3 = 9.5 Hz, H-2), 5.43 (dd, 1 H,
0
0
0
0
J4 ,5 = 0.9 Hz, J3 ,4 = 3.3 Hz, H-4G), 5.43 (t, 1 H, J2,3 = J3,4 = 9.5 Hz,
H-3), 5.25 (dd, 1 H, J3,4 = 9.5 Hz, J4,5 = 10.0 Hz, H-4), 5.21 (t, 1 H,
J1G,2G = J2G,3G = 10.0 Hz, H-2G), 5.05 (dd, 1 H, J3G,4G = 3.3 Hz, J3G,4G
=
10.0 Hz, H-3G), 4.70 (sa, 2 H, CH2Ar), 4.63 (d, 1 H, J1G,2G = 10.0 Hz,
H-1G), 4.30 (dd, 1 H, J5,6a = 5.0 Hz, J6a,6b = 12.7 Hz, H-6a), 4.16 (m, 2 H,
H-6a, H-6b), 4.11 (dd, 1 H, J5G,6bG = 7.5 Hz, J6aG,6bG = 11.3 Hz, H-6bG),
4.03 (ddd, 1 H, J5,6b = 1.9 Hz, J5,6a = 5.0 Hz, J4,5 = 10.0 Hz, H-5), 3.97
(ddd, 1 H, J4G,5G = 0.9 Hz, J5G,6aG = 6.5 Hz, J5G,6bG = 7.5 Hz, H-5G),
3.74ꢀ3.63 (m, 6 H, 3 CH2O), 2.99 (m, 1 H, J = 6.7 Hz, J = 7.1 Hz, Jgem
=
13.5 Hz, CHS), 2.80 (m, 1 H, J = 6.1 Hz, J = 7.1 Hz, Jgem = 13.5 Hz,
CHS), 2.15, 2.09, 2.07, 2.06, 2.05, 2.03, 1.99, 1.88 (8 s, 24 H, 8 CH3CO);
13C NMR (125 MHz, CDCl3) δ 170.4, 170.3, 170.1, 169.9, 169.8, 169.5,
169.2, 168.8 (ꢀCOCH3), 145.7 (C-4 triazole), 120.1 (C-5 triazole),
85.6 (C-1), 84.0 (C-1G), 75.0 (C-5), 74.3 (C-5G), 72.6 (C-3), 71.7
(C-3G), 71.1, 70.2, 70.1, 69.6 (C-2, 3 CH2O), 67.6 (C-4), 67.2 (ꢁ2) (C-
2G, C-4G), 64.3 (CH2Ar), 61.5, 61.3 (C-6, C-6G), 29.4 (CH2S), 20.7,
20.6 (ꢁ2), 20.5, 20.4 (ꢁ2), 20.1 (ꢁ2) (CH3CO). Anal. Calcd for
C35H49N3O20S: C, 48.66; H, 5.72; N, 4.86; S, 3.71. Found: C, 48.55; H,
5.74; N, 4.76; S, 4.02. HRMS (ESI) m/z [M þ Na]þ calcd for C35H49-
N3NaO20S 886.2528, found 886.2495.
Compound 13c: solvent system hexane/EtOAc (1:9); yield 72%;
[R]20D ꢀ12.1 (c 1.1, CHCl3); Rf 0.39 (EtOAc); 1H NMR (500 MHz,
CDCl3) δ 7.75 (s, 1 H, H-triazole), 5.83 (d, 1 H, J1,2 = 8.7 Hz, H-1),
5.41ꢀ5.32 (m, 3 H, H-2, H-3, H-4G), 5.17 (t, 1 H, J3,4 = J4,5 = 9.6 Hz,
H-4), 5.12 (t, 1 H, J1G,2G = J2G,3G = 10.0 Hz, H-2G), 4.98 (dd, 1 H,
J3G,4G = 3.3 Hz, J2G,3G = 10.0 Hz, H-3G), 4.63 (s, 2 H, CH2Ar), 4.53
Compound 11: solvent system hexane/EtOAc (2:1); yield 72%;
[R]20D ꢀ43.1 (c 0.6, CHCl3); Rf 0.35 (hexane/EtOAc 1:1); 1H NMR
(500 MHz, CDCl3) δ 7.49 (s, 1 H, H-triazole), 5.44 (dd, 1 H, J4,5 = 1.0
Hz, J3,4 = 3.4 Hz, H-4), 5.25 (t, 1 H, J1,2 = J2,3 = 10.0 Hz, H-2), 5.05 (dd,
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(d, 1 H, J1G,2G = 10.0 Hz, H-1G), 4.23 (dd, 1 H, J5,6a = 5.0 Hz, J6a,6b
12.6 Hz, H-6a), 4.09 (dd, 1 H, J5,6b = 2.1 Hz, J6a,6b = 12.6 Hz, H-6b), 4.08
(dd, 1 H, J5G,6aG = 6.5 Hz, J6aG,6bG = 11.2 Hz, H-6aG), 4.04 (dd, 1 H,
=
C34H49N3NaO19S 858.2573, found 858.2548; m/z [M þ H]þ calcd for
C34H50N3O19S 836.2754, found 836.2743.
Compound 15c: solvent system EtOAc/MeOH (98:2); yield 75%;
1
[R]20 þ45.8 (c 0.3, CHCl3); Rf 0.2 (hexane/EtOAc 1:6); H NMR
J
5G,6bG = 6.5 Hz, J6aG,6bG = 11.3 Hz, H-6bG), 3.96 (ddd, 1 H, J5,6b = 2.0
D
Hz, J5,6a = 5.2 Hz, J4,5 = 10.0 Hz, H-5), 3.89 (br dd, 1 H, J4G,5G < 1.0 Hz,
J5G,6aG = 6.5 Hz, J5G,6bG = 6.7 Hz, H-5G), 3.65ꢀ3.54 (m, 18 H, 9
CH2O), 2.90, 2.73 (2 m, 2 H, J = 6.8 Hz, Jgem = 13.4 Hz, CH2S), 2.09,
2.02, 2.01, 2.00, 1.98, 1.96, 1.91, 1.80 (8 s, 24 H, 8 CH3CO); 13C NMR
(125 MHz, CDCl3) δ 170.8, 170.7, 170.6, 170.3, 170.2, 169.9, 169.7,
169.2 (ꢀCOCH3), 146.3 (C-4 triazole), 121.4 (C-5 triazole), 86.0 (C-
1), 84.5 (C-1G), 75.4 (C-5), 74.8 (C-5G), 73.1 (C-3), 72.2 (C-3G), 71.5
(C-2), 70.8 (ꢁ6), 70.7, 70.6, 70.1 (9 CH2O), 68.1 (C-4), 67.8 (C-2G),
67.7 (C-4G), 64.8 (CH2Ar), 62.0, 61.9 (C-6, C-6G), 29.9 (CH2S), 21.1,
21.0 (ꢁ2), 20.9, 20.8, 20.5 (ꢁ3) (ꢀCOCH3). Anal. Calcd for
C41H61N3O23S: C, 49.44; H, 6.17; N, 4.22; S, 3.22. Found: C, 49.59;
H, 6.09; N, 4.30; S, 3.46. HRMS (ESI) m/z [M þ Na]þ calcd for
C41H61N3NaO23S 1018.3314, found 1018.3353.
(500 MHz, CDCl3) δ 7.65 (sa, 1 H, H-triazole), 5.41 (dd, 1 H, J3,4 = 9.5
Hz, J2,3 = 10.0 Hz, H-3), 5.35 (dd, 1 H, J4G,5G = 0.7 Hz, J3G,4G = 3.3 Hz,
H-4G), 5.13 (t, 1 H, J1G,2G = J2G,3G = 10.0 Hz, H-2G), 4.97 (dd, 1 H,
J3G,4G = 3.4 Hz, J2G,3G = 10.0 Hz, H-3G), 4.83 (d, 1 H, J1,2 = 3.6 Hz,
H-1), 4.78 (dd, 1 H, J3,4 = 9.5 Hz, J4,5 = 10.0 Hz, H-4), 4.76 (dd, 1 H,
J1,2 = 3.6 Hz, J2,3 = 10.2 Hz, H-2), 4.62 (m, 2 H, CH2Ar), 4.52 (d, 1 H,
J1G,2G = 10.0 Hz, H-1G), 4.51 (m, 1 H, J5,6a = 2.0 Hz, H-6a), 4.30 (dd,
1 H, J5,6b = 8.6 Hz, J6a,6b = 14.2 Hz, H-6b), 4.12 (ddd, 1 H, J5,6a = 2.0 Hz,
J5,6b = 8.6 Hz, J4,5 = 10.0 Hz, H-5), 4.07 (dd, 1 H, J5G,6aG = 6.7 Hz,
J6aG,6bG = 11.5 Hz, H-6aG), 4.03 (dd, 1 H, J5G,6bG = 6.5 Hz, J6aG,6bG
=
11.4 Hz, H-6bG), 3.87 (ddd, 1 H, J4G,5G = 0.7 Hz, J5G,6aG = 6.7 Hz,
J5G,6bG = 6.5 Hz, H-5G), 3.65ꢀ3.55 (m, 18 H, 9 CH2O), 3.05 (s, 3 H,
ꢀOCH3), 2.89 (m, 1 H, J = 6.7 Hz, J = 6.9 Hz, Jgem = 13.5 Hz, CHS),
2.73 (m, 1 H, J = 6.1 Hz, J = 7.2 Hz, Jgem = 13.5 Hz, CHS), 2.08, 2.03,
1.99, 1.98, 1.97, 1.94, 1.91 (7 s, 21 H, 7 CH3CO); 13C NMR (125 MHz,
CDCl3 þ DMSO) δ 170.3, 170.2, 170.1, 170.0, 169.8 (ꢁ2), 169.5, 169.6
(ꢀCOCH3), 144.5 (C-4 triazole), 124.2 (C-5 triazole), 96.3 (C-1), 83.8
(C-1G), 74.3 (C-5G), 71.5 (C-3G), 71.8, 71.1, 70.7, 70.6, 70.5, 70.4,
70.3, 70.2, 70.0, 69.7 (C-2, C-3, C-4, 9 ꢁ CH2O), 67.7 (C-5), 67.3 (C-
4G), 67.2 (C-2G), 64.5 (CH2Ar), 61.4 (C-6G), 55.4 (ꢀOCH3), 50.8
(C-6), 29.4 (CH2S), 20.8, 20.6 (ꢁ5), 20.5 (ꢀCOCH3). Anal. Calcd for
Compound 15a: solvent system hexane/EtOAc (2:3); yield 75%;
mp 88ꢀ90 °C; [R]20D þ20.1 (c 0.4, CHCl3); Rf 0.34 (hexane/EtOAc
1:2); 1H NMR (500 MHz, CDCl3) δ 7.62 (s, 1 H, H-triazole), 5.48 (dd,
1 H, J3,4 = 9.4 Hz, J2,3 = 10.2 Hz, H-3), 5.43 (dd, 1 H, J4G,5G = 1.0 Hz,
J
3G,4G = 3.4 Hz, H-4G), 5.26 (t, 1 H, J1G,2G = J2G,3G = 10.0 Hz, H-2G),
5.04 (dd, 1 H, J3G,4G = 3.4 Hz, J2G,3G = 10.0 Hz, H-3G), 4.92 (d, 1 H,
1,2 = 3.6 Hz, H-1), 4.81 (dd, 1 H, J1,2 = 3.6 Hz, J2,3 = 10.2 Hz, H-2), 4.76
J
(dd, 1 H, J3,4 = 9.4 Hz, J4,5 = 10.1 Hz, H-4), 4.57 (d, 1 H, J1G,2G = 10.0 Hz,
H-1G), 4.57 (dd, 1 H, J5,6a = 2.4 Hz, J6a,6b = 14.5 Hz, H-6a), 4.42 (dd,
1 H, J5,6b = 7.9 Hz, J6a,6b = 14.5 Hz, H-6b), 4.21 (ddd, 1 H, J5,6a = 2.4 Hz,
C40H61N3O22S 0.5H2O: C, 49.17; H, 6.40; N, 4.30; S, 3.28. Found: C,
3
49.17; H, 6.32; N, 3.99; S, 3.11. HRMS (ESI) m/z [M þ Na]þ calcd for
C40H61N3NaO22S 990.3360, found 990.3352; m/z [M þ H]þ calcd for
C40H62N3O22S 968.3540, found 868.3539.
J
5,6b = 7.9 Hz, J4,5 = 10.1 Hz, H-5), 4.13 (m, 3 H, H-6aG, H-6bG, CHS),
3.96 (ddd, 1 H, J4G,5G = 1.0 Hz, J5G,6aG = J5G,6bG = 6.5 Hz, H-5G), 3.91
(d, 1 H, Jgem = 14.3 Hz, CHS), 3.18 (s, 3 H, ꢀOCH3), 2.16, 2.10, 2.06
(ꢁ2), 2.05, 2.01, 1.97 (7 s, 21 H, 7 CH3CO); 13C NMR (125 MHz,
CDCl3) δ 170.4, 170.2, 170.1, 169.9 (ꢁ2), 169.8, 169.6 (ꢀCOCH3),
144.8 (C-4 triazole), 123.5 (C-5 triazole), 96.7 (C-1), 82.9 (C-1G), 74.4
(C-5G), 71.7 (C-3G), 70.7 (C-2), 69.7 (ꢁ2) (C-4, C-3), 67.6 (C-5),
67.3, 67.2 (C-2G, C-4G), 61.3 (C-6G), 55.5 (ꢀOCH3), 50.7 (C-6), 24.1
(CH2S), 20.7 (ꢁ3), 20.6 (ꢁ3) (ꢀCOCH3). Anal. Calcd for
C30H41N3O17S: C, 48.19; H, 5.53; N, 5.62; S, 4.29. Found: C, 47.95;
H, 5.68; N, 5.56; S, 4.07. HRMS (ESI) m/z [M þ Na]þ calcd for
C30H41N3NaO17S 770.2054, found 770.2044.
Compound 15b: solvent system hexane/EtOAc (1:9); yield 79%;
[R]20D þ42.4 (c 0.4, CHCl3); Rf 0.18 (hexane/EtOAc 1:2); 1H NMR
(500 MHz, CDCl3) δ 7.64 (s, 1 H, H-triazole), 5.41 (dd, 1 H, J3,4 = 9.4
Hz, J2,3 = 10.1 Hz, H-3), 5.33 (dd, 1 H, J4G,5G = 0.9 Hz, J3G,4G = 3.4 Hz,
H-4G), 5.13 (t, 1 H, J1G,2G = J2G,3G = 10.0 Hz, H-2G), 4.96 (dd, 1 H,
J3G,4G = 3.4 Hz, J2G,3G = 10.0 Hz, H-3G), 4.83 (d, 1 H, J1,2 = 3.6 Hz,
H-1), 4.78 (dd, 1 H, J3,4 = 9.4 Hz, J4,5 = 10.1 Hz, H-4), 4.76 (dd, 1 H,
J1,2 = 3.6 Hz, J2,3 = 10.2 Hz, H-2), 4.63 (m, 2 H, CH2Ar), 4.55 (d, 1 H,
J1G,2G = 10.0 Hz, H-1G), 4.52 (dd, 1 H, J5,6a = 2.5 Hz, J6a,6b = 14.3 Hz,
H-6a), 4.31 (dd, 1 H, J5,6b = 8.5 Hz, J6a,6b = 14.4 Hz, H-6b), 4.13 (ddd, 1
H, J5,6a = 2.4 Hz, J5,6b = 8.7 Hz, J4,5 = 10.1 Hz, H-5), 4.07 (dd, 1 H,
Compound 17a: Solvent system, hexane/EtOAc (1:9); yield 60%;
mp 125ꢀ127 °C; [R]20 ꢀ26.8 (c = 1.0, CHCl3); Rf 0.17 (hexane/
D
1
EtOAc 1:2); H NMR (500 MHz, CDCl3) δ 7.82, 7.41 (2 s, 2 H,
H-triazole), 5.89 (d, 1 H, J1,2 = 8.9 Hz, H-1), 5.51ꢀ5.45 (m, 3 H, H-2,
0
0
0
0
0
H-3, H-4G*), 5.44 (dd, 1 H, J4G ,5G = 0.9 Hz, J3G ,4G = 3.3 Hz, H-4G *),
5.27ꢀ5.19 (m, 3 H, H-2G, H-2G0, H-4), 5.17 (dd, 1 H, J3G ,4G = 3.4 Hz,
0
0
0
0
0
J2G ,3G = 9.9 Hz, H-3G *), 5.07 (dd, 1 H, J3G,4G = 3.4 Hz, J2G,3G = 10.0
Hz, H-3G*), 4.66 (dd, 1 H, J5,6a = 2.5 Hz, J6a,6b = 14.8 Hz, H-6a), 4.58 (d,
1 H, J1G,2G = 10.0 Hz, H-1G), 4.51 (dd, 1 H, J5,6b = 7.0 Hz, J6a,6b = 14.8
0
0
0
0
Hz, H-6b), 4.42 (dd, 1 H, J5G ,6aG = 4.4 Hz, J6aG ,6bG = 11.0 Hz,
H-6aG0), 4.29 (d, 1 H, J1G ,2G = 9.9 Hz, H-1G ), 4.26 (ddd, 1 H, J5,6a
=
0
0
0
2.5 Hz, J5,6b = 7.0 Hz, J4,5 = 9.8 Hz, H-5), 4.16 (d, 1 H, Jgem = 14.0 Hz,
CHS), 4.10 (dd, 1 H, J5G,6aG = 6.5 Hz, J6aG,6bG = 11.2 Hz, H-6aG),
4.08ꢀ4.03 (m, 3 H, CHS, H-5G0, H-6bG0), 3.98 (dd, 1 H, J5G ,6bG = 7.4
0
0
0
0
0
Hz, J6aG ,6bG = 11.0 Hz, H-6bG ), 3.91 (ddd, 1 H, J4G,5G = 0.9 Hz, J5G,6aG
≈ J5G,6bG = 6.5 Hz, H-5G), 3.89 (d, 1 H, Jgem = 14.1 Hz, CHS*), 3.87 (d,
1 H, Jgem = 14.0 Hz, CHS0*), 2.20, 2.16, 2.15, 2.14, 2.06, 2.04, 2.01, 1.99,
1.98, 1.97, 1.90 (11 s, 33 H, 11 CH3CO); 13C NMR (125 MHz, CDCl3) δ
171.6, 170.5, 170.3, 170.1, 170.0 (ꢁ2), 169.8, 169.7 (ꢁ2), 169.6, 169.4
(ꢀCOCH3), 144.9, 143.7 (C-4 triazole), 123.4, 121.3 (C-5 triazole), 85.8
(C-1), 83.2 (C-1G), 80.4 (C-1G0), 75.3 (C-5), 74.5 (C-5G), 73.4 (C-5G0),
72.1 (C-3), 71.7, 71.6 (C-3G, C-3G0), 72.3 (C-2), 68.8 (C-4), 67.6, 67.4,
67.3 (ꢁ2) (C-2G, C-2G0, C-4G, C-4G0), 61.3, 61.1 (C-6G, C-6G0), 50.5
(C-6), 24.2, 23.2 (2 ꢁ CH2S), 20.8 (ꢁ3), 20.7 (ꢁ3), 20.6 (ꢁ3), 20.2
(ꢁ2) (ꢀCH3CO). Anal. Calcd for C46H60N6O25S2: C, 47.58; H, 5.21; N,
7.24; S, 5.52. Found: C, 47.94; H, 5.11; N, 7.15; S, 5.31. HRMS (ESI) m/z
[M þ Na]þ calcd for C46H60N6NaO25S2 1183.2947, found 1183.2968.
J5G,6aG = 6.8 Hz, J6aG,6bG = 11.3 Hz, H-6aG), 4.02 (dd, 1 H, J5G,6bG
=
6.6 Hz, J6aG,6bG = 11.3 Hz, H-6bG), 3.88 (ddd, 1 H, J4G,5G = 1.0 Hz,
J5G,6aG = 6.8 Hz, J5G,6bG = 6.6 Hz, H-5G), 3.67ꢀ3.55 (m, 6 H, 3 CH2O),
3.05 (s, 3 H, ꢀOCH3), 2.91 (m, 1 H, J = 7.0 Hz, J = 6.8 Hz, Jgem = 13.6
Hz, CHS), 2.72 (m, 1 H, J = 6.1 Hz, J = 6.9 Hz, Jgem = 13.5 Hz, CHS),
2.08, 2.03, 1.99 (ꢁ2), 1.98, 1.94, 1.91 (7 s, 21 H, 7 CH3CO); 13C NMR
(125 MHz, CDCl3) δ 170.3, 170.2, 170.1, 170.0, 169.8 (ꢁ2), 169.5,
169.6 (ꢀCOCH3), 145.0 (C-4 triazole), 124.4 (C-5 triazole), 96.5 (C-
1), 84.0 (C-1G), 74.3 (C-5G), 71.8 (C-3G), 71.2 (CH2O), 70.7 (C-2),
70.2 (CH2O), 70.0 (C-4), 69.7 (C-3), 69.6 (CH2O), 67.7 (C-5), 67.3
(C-4G), 67.3 (C-2G), 64.4 (CH2Ar), 61.4 (C-6G), 55.4 (ꢀOCH3), 50.7
(C-6), 29.4 (CH2S), 20.8, 20.6 (ꢁ5), 20.5 (ꢀCOCH3). Anal. Calcd for
Compound 17b: solvent system hexane/EtOAc (1:9); yield 84%;
1
[R]20 ꢀ4.6 (c 1.0, CHCl3); Rf 0.32 (EtOAc); H NMR (500 MHz,
D
CDCl3) δ 7.80, 7.56 (2 s, 2 H, H-triazole), 5.86 (d, 1 H, J1,2 = 9.2 Hz,
H-1), 5.50 (t, 1 H, J1,2 = 9.2 Hz, J2,3 = 9.4 Hz, H-2), 5.43 (t, 1 H, J3,4 = 9.3
Hz, J2,3 = 9.4 Hz, H-3), 5.43 (dd, 1 H, J4G,5G = 0.9 Hz, J3G,4G =0 3.4 Hz,
0
0
0
0
H-4G*), 5.39 (dd, 1 H, J4G ,5G = 1.0 Hz, J3G ,4G = 3.4 Hz, H-4G *), 5.21
0
0
C34H49N3O19S 0.5H2O: C, 48.34; H, 5.97; N, 4.97; S, 3.80. Found: C,
(t, 1 H, J1G,2G = 0.9 Hz, J2G,3G = 10.0 Hz, H-2G*), 5.18 (t, 1 H, J1G ,2G
=
3
48.01; H, 5.74; N, 4.93; S, 3.64. HRMS (ESI) m/z [M þ Na]þ calcd for
J2G ,3G = 10.0 Hz, H-2G *), 5.09 (dd, 1 H, J3,4 = 9.3 Hz, J4,5 = 10.1 Hz,
0
0
0
3071
dx.doi.org/10.1021/jo102421e |J. Org. Chem. 2011, 76, 3064–3077

The Journal of Organic Chemistry
ARTICLE
H-4), 5.06 (dd, 1 H, J3G,4G = 3.4 Hz, J2G,3G =10.0 Hz, H-3G*), 5.03 (dd,
(C-3), 69.4 (C-2), 69.3 (C-5), 67.7 (ꢁ2) (C-2G, C-4G), 61.8 (C-6G),
51.0 (C-6), 24.5 (CH2S), 21.2, 21.1, 21.0, 20.9, 20.8, 20.7, 20.5
(ꢀCOCH3). Anal. Calcd for C58H76N6O33S2: C, 48.06; H, 5.29; N,
5.80; S, 4.42. Found: C, 48,02; H, 5,17; N, 5.55; S, 4.58. HRMS (ESI) m/z
[M þ Na]þ calcd for C58H76N6NaO33S2 1471.3792, found 1471.3796.
Compound 19b: solvent system EtOAc/MeOH (99:2); yield 68%;
[R]20D þ32.9 (c 0.2, CHCl3); Rf 0.42 (EtOAc); 1H NMR (500 MHz,
CDCl3) δ 7.61 (s, 1 H, H-triazole), 5.42 (t, 1 H, J2,3 = J3,4 = 9.7 Hz, H-3),
5.37 (br d, 1 H, J4G,5G < 1.0 Hz, J3G,4G = 2.7 Hz, H-4G), 5.17 (t, 1 H,
0
0
0
0
0
1 H, J3G ,4G = 3.4 Hz, J2G ,3G =10.0 Hz, H-3G *), 4.70 (s, 2 H, CH2Ar),
4.67 (dd, 1 H, J5,6a = 2.6 Hz, J6a,6b =14.9 Hz, H-6a), 4.66 (d, 1 H, J1G,2G
=
0
0
10.0 Hz, H-1G), 4.65 (s, 2 H, CH2Ar), 4.64 (d, 1 H, J1G ,2G = 10.0 Hz,
H-1G0), 4.49 (dd, 1 H, J5,6b = 7.6 Hz, J6a,6b = 14.9 Hz, H-6b), 4.29 (ddd, 1
H, J5,6a = 2.6 Hz, J5,6b = 7.6 Hz, J4,5 = 10.2 Hz, H-5), 4.18ꢀ4.08 (m, 4 H,
H-6aG, H-6bG, H-6aG0, H-6bG0), 3.98 (ddd, 1 H, J4G,5G = 0.9 Hz,
0
0
J5G,6aG = J5G,6bG = 6.6 Hz, H-5G), 3.96 (ddd, 1 H, J4G ,5G = 0.9 Hz,
J5G ,6aG = J5G ,6bG = 6.6 Hz, H-5G0), 3.76ꢀ3.60 (m, 12 H, 6 CH2O),
3.02ꢀ2.94 (m, 2 H, J = 6.9 Hz, J = 10.5 Hz, Jgem = 13.7 Hz, 2 ꢁ CHS),
2.84ꢀ2.74 (m, 2 H, J = 6.1 Hz, J = 7.1 Hz, J = 13.7 Hz, 2 ꢁ CHS), 2.16,
2.15, 2.13, 2.06, 2.05, 2.04 (ꢁ2), 2.03, 1.99, 1.98, 1.87 (11 s, 33 H, 11
CH3CO); 13C NMR (125 MHz, CDCl3) δ 171.1, 170.4, 170.2 (ꢁ2),
170.0 (ꢁ2), 169.8, 169.6, 169.5, 168.7 (CH3CO), 145.8, 145.2 (C-4
triazole), 124.0, 121.1 (C-5 triazole), 85.4 (C-1), 84.1, 84.0 (C-1G,
C-1G0), 75.4 (C-5), 74.3 (ꢁ2) (C-5G, C-5G0), 72.4 (C-3), 71.8 (ꢁ2)
(C-3G, C-3G0), 71.2 (ꢁ2), 70.2, 70.1, 70.0, 69.8 (6 CH2O), 69.0 (C-4),
67.4, 67.3 (ꢁ2), 67.2 (C-2G, C-2G0, C-4G, C-4G0), 64.4, 64.3 (2
CH2Ar), 64.5, 61.4 (C-6G, C-6G0), 50.4 (C-6), 29.5 (ꢁ2) (CH2S),
21.0, 20.8, 20.6 (ꢁ2), 20.4, 20.1 (CH3CO). Anal. Calcd for
C54H76N6O29S2: C, 48.50; H, 5.73; N, 6.28; S, 4.80. Found: C, 48.24;
H, 5.61; N, 6.54; S, 4.50. HRMS (ESI) m/z [M þ Na]þ calcd for
C54H76N6NaO29S2 1359.3990, found 1359.3973.
0
0
0
0
J1G,2G = J2G,3G = 10.0 Hz, H-2G), 4.98 (dd, 1 H, J3G,4G = 2.8 Hz, J2G,3G
=
10.0 Hz, H-3G), 4.97 (dd, 1 H, J1,2 = 3.3 Hz, J2,3 = 10.3 Hz, H-2), 4.88 (t,
1 H, J3,4 = J4,5 = 9.6 Hz, H-4), 4.82 (d, 1 H, J1,2 = 3.1 Hz, H-1), 4.66 (s, 2
H, CH2Ar), 4.62 (d, 1 H, J1G,2G = 10.0 Hz, H-1G), 4.52 (dd, 1 H, J5,6a
<
1.0 Hz, J6a,6b = 13.4 Hz, H-6a), 4.31 (dd, 1 H, J5,6b = 8.6 Hz, J6a,6b = 13.4
Hz, H-6b), 4.24 (ddd, 1 H, J5,6a < 1.0 Hz, J5,6b = 8.6 Hz, J4,5 = 9.6 Hz,
H-5), 4.12 (dd, 1 H, J5G,6aG = 6.7 Hz, J6aG,6bG = 11.2 Hz, H-6aG), 4.07
(dd, 1 H, J5G,6bG = 6.6 Hz, J6aG,6bG = 11.2 Hz, H-6bG), 3.94 (ddd, 1 H,
J4G,5G < 1.0 Hz, J5G,6aG = J5G,6bG = 6.5 Hz, H-5G), 3.66 (m, 6 H, 3
CH2O), 2.97, 2.77 (2 t, 2 H, J = 6.8 Hz, Jgem = 13.5 Hz, CH2S), 2.13, 2.08,
2.04, 2.03, 2.02, 1.98, 1.96 (7 s, 21 H, 7 CH3CO); 13C NMR (125 MHz,
CDCl3) δ 170.7, 170.6, 170.4, 170.2, 170.1, 170.0, 169.8 (ꢀCOCH3),
145.8 (C-4 triazole), 124.4 (C-5 triazole), 92.1 (C-1), 84.4 (C-1G), 74.8
(C-5G), 72.2 (C-3G), 71.6, 70.5 (CH2O), 70.2 (C-4), 70.1 (C-3), 70.0
(3 CH2O), 69.4 (C-5), 69.1 (C-2), 67.8 (C-2G), 67.7 (C-4G), 64.8
(CH2Ar), 61.9 (C-6G), 51.0 (C-6), 29.9 (CH2S), 21.2, 21.1, 21.0, 20.9,
20.8, 20.7, 20.6 (ꢀCOCH3). Anal. Calcd for C66H92N6O37S2: C, 48.76;
H, 5.70; N, 5.17; S, 3.95. Found: C, 48.46; H, 5.61; N, 5.25; S, 4.09.
HRMS (ESI) m/z [M þ Na]þ calcd for C66H92N6NaO37S2 1647.4841,
found 1647.4764.
Compound 19c: solvent system EtOAc/MeOH (95:5); yield 78%;
[R]20D þ38.1 (c 0.7, CHCl3); Rf 0.49 (EtOAc/MeOH 9:1); 1H NMR
(500 MHz, DMSO-d6) δ 7.98 (s, 1 H, H-triazole), 5.32 (br d, 1 H, J4G,5G
< 1.0 Hz, J3G,4G = 3.2 Hz, H-4G), 5.28 (t, 1 H, J2,3 = J3,4 = 9.8 Hz, H-3),
5.17 (dd, 1 H, J3G,4G = 3.4 Hz, J2G,3G = 9.7 Hz, H-3G), 5.02 (dd, 1 H,
J1,2 = 3.4 Hz, J2,3 = 10.2 Hz, H-2), 4.99 (t, 1 H, J3,4 = J4,5 = 9.9 Hz, H-4),
4.97 (t, 1 H, J1G,2G = J2G,3G = 9.6 Hz, H-2G), 4.91 (d, 1 H, J1G,2G = 10.0
Hz, H-1G), 4.83 (d, 1 H, J1,2 = 3.2 Hz, H-1), 4.63 (dd, 1 H, J5,6a < 1.0 Hz,
J6a,6b = 13.3 Hz, H-6a), 4.51 (m, 3 H, H-6b, CH2Ar), 4.22 (m, 2 H, H-5,
H-5G), 4.04 (m, 2 H, H-6aG, H-6bG), 3.52 (m, 18 H, 9 CH2O), 2.84,
2.77 (2 m, 2 H, J0 = 6.7, Jgem = 13.4 Hz, CH2S), 2.13, 2.08, 2.03, 2.01, 2.00,
1.96, 1.93(7s, 21H, 7CH3CO); 13C NMR (125 MHz, DMSO-d6) δ170.9,
170.8, 170.7, 170.6, 170.3, 170.2, 169.9 (COCH3), 145.0 (C-4 triazole),
125.8 (C-5 triazole), 91.8 (C-1), 83.6 (C-1G), 74.3 (C-5G), 72.9 (C-3G),
71.1, 70.6, 70.5, 70.4, 70.1, 69.8 (C-3, C-4, 9 CH2O), 69.4 (C-5), 69.0 (C-
2), 68.5 (C-4G), 68.1 (C-2G), 64.2 (CH2Ar), 62.5 (C-6G), 50.5 (C-6),
30.1 (CH2S), 21.4, 21.3, 21.2, 21.1, 20.8, 20.7, 20.6 (ꢀCOCH3). Anal.
Calcd for C78H116N6O43S2: C, 49.57; H, 6.19; N, 4.45; S, 3.39. Found: C,
49.79; H, 6.15; N, 4.33; S, 3.59. HRMS (ESI) m/z [M þ Na]þ calcd for
C78H116N6NaO43S2 1911.6414, found 1911.6376.
Compound 17c: solvent system hexane/EtOAc (6:94); yield 76%;
[R]20 ꢀ0.4 (c 1.0, CHCl3); [R]20D þ5.1 (c 0.9, DMSO-d6); Rf 0.11
D
(EtOAc/MeOH 1.9:1); 1H NMR (500 MHz, DMSO-d6) δ 8.36, 7.93 (2
s, 2 H, H-triazole), 6.30 (d, 1 H, J1,2 = 9.2 Hz, H-1), 5.66 (t, 1 H, J1,2 = J2,3
= 9.3 Hz, H-2), 5.55 (t, 1 H, J2,3 = J3,4 = 9.4 Hz, H-3), 5.33 (m, 2 H,
H-4G, H-4G0), 5.18 (dd, 2 H, J3G,4G = J3G ,4G = 3.4 Hz, J2G,3G = J2G ,3G
=
0
0
0
0
9.7 Hz, H-3G, H-3G0), 5.11 (t, 1 H, J3,4 = J4,5 = 9.4 Hz, H-4), 4.99 (t, 2 H,
0
0
0
0
0
J1G,2G = J1G ,2G = J2G,3G = J2G ,3G = 9.80Hz, H-2G, H-2G ), 4.91 (d, 2 H,
0
0
J
1G,2G = J1G ,2G = 10.0 Hz, H-1G, H-1G ), 4.67 (m, 2 H, H-5, H-6a), 4.54
(m, 5 H, H-6b, 2 ꢁ CH2Ar), 4.23 (m, 2 H, H-5, H-5G), 4.02 (m, 2 H,
H-6aG, H-6aG0, H-6bG, H-6bG0), 3.55ꢀ3.49 (m, 36 H, 18 CH2O),
2.84, 2.74 (2 m, 4 H, J = 6.7 Hz, Jgem = 13.5, 2 CH2S), 2.13 (ꢁ2), 2.06,
2.03 (ꢁ2), 2.01 (ꢁ2), 1.96, 1.93 (ꢁ2), 1.80 (11 s, 33 H, 11 CH3CO);
13C NMR (125 MHz, DMSO-d6) δ 170.9 (ꢁ2), 170.8 (ꢁ2), 170.4,
170.3 (ꢁ2), 170.2, 170.1, 170.0, 169.4 (ꢀCOCH3), 145.6, 144.9 (C-4
triazole), 125.7, 123.6 (C-5 triazole), 84.7 (C-1), 83.6 (ꢁ2) (C-1G,
C-1G0), 74.5 (C-5), 74.3 (ꢁ2) (C-5G, C-5G0), 73.0 (C-3), 71.9 (ꢁ2)
(C-3G, C-3G0), 71.1 (ꢁ2), 71.0, 70.6 (ꢁ4), 70.5 (ꢁ2), 70.4 (ꢁ2), 70.1,
69.9 (ꢁ2), 69.8 (ꢁ2), 69.7 (ꢁ2) (C-2, C-4, 18 CH2O), 68.5 (ꢁ2) (C-
4G, C-4G0), 68.1 (ꢁ2) (C-2G, C-2G0), 64.3, 64.2 (2 ꢁ CH2Ar), 62.5
(ꢁ2) (C-6G, C-6G0), 50.8 (C-6), 30.1 (ꢁ2) (2 CH2S), 21.4 (ꢁ2), 21.3
(ꢁ2), 21.2 (ꢁ2), 21.1 (ꢁ2), 21.0 (ꢁ2), 20.7 (ꢀCOCH3). Anal. Calcd
for C66H100N6O35S2: C, 49.49; H, 6.29; N, 5.25; S, 4.00. Found: C,
49.70; H, 6.14; N, 4.95; S, 4.25. HRMS (ESI) m/z [M þ Na]þ calcd for
C66H100N6NaO35S2 1623.5569, found 1623.5592.
Compound 19a: solvent system hexane/EtOAc (2:3); yield 69%;
mp 112ꢀ114 °C; [R]20D þ5.8 (c 1.0, CHCl3); Rf 0.17 (hexane/EtOAc
1:4); 1H NMR (500 MHz, CDCl3) δ 7.46 (s, 1 H, H-triazole), 5.39 (t, 1
H, J2,3 = J3,4 = 9.7 Hz, H-3), 5.37 (br d, 1 H, J4G,5G < 1.0 Hz, J3G,4G = 3.3
Hz, H-4G), 5.18 (t, 1 H, J1G,2G = J2G,3G = 10.0 Hz, H-2G), 4.98 (dd, 1 H,
J3G,4G = 3.3 Hz, J2G,3G = 10.0 Hz, H-3G), 4.90 (dd, 1 H, J1,2 = 3.7 Hz, J2,3
Compound 21a: solvent system cyclohexane/EtOAc (85:15);
yield 54%; mp 120ꢀ122 °C; [R]20 ꢀ8.7 (c 0.7, CHCl3); Rf 0.24
D
1
(hexane/EtOAc 1:6); H NMR (500 MHz, DMSO-d6) δ 8.11, 7.93,
7.81 (3s, 3 H, H-triazole), 6.26 (d, 1 H, J1,2 = 9.1 Hz, H-1), 5.62 (t, 1 H,
0
0
0
J2,3 = J3,4 = 9.1 Hz, H-3), 5.46 (d, 1 H, J1 ,2 = 3.6 Hz, H-1 ), 5.44 (t, 1 H,
J1,2 = J2,3 = 9.3 Hz, H-2), 5.33 (m, 4 H, H-30, 3 ꢁ H-4G), 5.19 (m, 3 H,
3 ꢁ H-3G), 5.01 (m, 3 H, J1G,2G = J2G,3G = 9.9 Hz, 3 ꢁ H-2G), 4.92 (dd,
= 10.1 Hz, H-2), 4.86 (d, 1 H, J1,2 = 3.7 Hz, H-1), 4.77 (t, 1 H, J3,4 = J4,5
9.6 Hz, H-4), 4.51 (d, 1 H, J1G,2G = 10.0 Hz, H-1G), 4.43 (dd, 1 H, J5,6a
=
=
1 H, J1 ,2 = 3.6 Hz, J2 ,3 = 10.4 Hz, H-20), 4.87, 4.82, 4.66 (3d, 3 H,
0
0
0
0
0.8 Hz, J6a,6b = 14.0 Hz, H-6a), 4.27 (dd, 1 H, J5,6b = 8.3 Hz, J6a,6b = 13.8
Hz, H-6b), 4.19 (ddd, 1 H, J5,6a = 0.8 Hz, J5,6b = 8.6 Hz, J4,5 = 9.9 Hz,
H-5), 4.11ꢀ4.02 (m, 3 H, H-6aG, H-6bG, CHS), 3.89 (ddd, 1 H,
J1G,2G = 9.9 Hz, 3 ꢁ H-1G), 4.82 (t, 1 H, J3 ,4 = J4 ,5 = 9.4 Hz, H-40),
4.73ꢀ4.62 (m, 2 H, H-60a, H-60b), 4.61ꢀ4.45 (m, 3 H, H-6a, H-5, H-50),
4.33 (dd, 1 H, J5,6b = 6.9 Hz, J6a,6b = 13.8 Hz, H-6b), 4.23ꢀ4.15 (m, 3 H,
3 ꢁ H-5G), 4.11ꢀ3.86 (m, 13 H, H-4, 3 ꢁ CH2S, 3 ꢁ H-6aG, 3 ꢁ
H-6bG), 2.13, 2.04, 2.02, 2.01, 1.99, 1.98, 1.97, 1.96, 1.93, 1.92, 1.76
(11s, 51 H, 17 ꢁ CH3CO); 13C NMR (125 MHz, CDCl3) δ 170.9,
170.8, 170.7, 170.4, 170.3, 170.2, 170.1, 170.0, 169.8 (CH3CO), 144.7,
0
0
0
0
J4G,5G < 1.0 Hz, J5G,6aG = J5G,6bG = 6.5 Hz, H-5G), 3.82 (d, 1 H, Jgem
=
14.2 Hz, CHS), 2.10, 2.04, 1.99, 1.98, 1.97, 1.94, 1.91 (7 s, 21 H,
7 CH3CO); 13C NMR (125 MHz, CDCl3) δ 170.8, 170.7, 170.4, 170.2,
170.1, 170.0, 169.9 (COCH3), 145.5 (C-4 triazole), 123.8 (C-5 triazole),
92.3 (C-1), 83.3 (C-1G), 74.9 (C-5G), 72.2 (C-3G), 70.1 (C-4), 70.0
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ARTICLE
144.4, 144.0 (C-4 triazole), 125.4, 125.1, 122.9 (C-5 triazole), 96.6 (C-
10), 84.3 (C-1), 82.8, 82.7, 82.4 (C-1G), 75.1, 75.0 (C-4, C-5), 74.8 (C-
3), 74.5, 74ꢀ4, 74ꢀ3 (C-5G), 71.9, 71.8, 71.7 (C-3G), 71.6 (C-2), 70.1
(C-20), 69.8 (C-30), 69.7 (C-40), 69.4 (C-50), 68.4, 68.3, 68.2, 68.1, 68.0
(C-2G, C-4G), 62.4, 62.2, 62.1 (C-6G), 50.9, 50.4 (C-6, C-60), 24.5,
24.4, 24.1 (3 ꢁ CH2S), 21.3, 21.2, 21.1 (CH3CO). Anal. Calcd for
C73H95N9O40S3: C, 47.79; H, 5.22; N, 6.87; S, 5.24. Found: C, 47.58; H,
5.27; N, 6.69; S, 5.44. HRMS (ESI) m/z [M þ Na]þ calcd for
C73H95N9NaO40S3 1856.4736, found 1856.4706.
(18s, 69 H, 23 ꢁ CH3CO); 13C NMR (125 MHz, DMSO-d6) δ 170.4,
170.3, 170.2, 170.1, 170.0, 169.9, 169.8, 169.7, 169.5, 169.4 (CH3CO),
144.2, 144.0, 143.5 (C-4 triazole), 125.8, 125.4, 125.1, 122.6 (C-5
triazole), 96.1, 95.9 (C-10, C-100), 84.0 (C-1), 82.5, 82.3 (ꢁ2), 81.9
(C-1G), 75.6, 74.7, 74.3, 74.0, 73.9, 73.7, 71.5, 71.0, 70.3, 69.5, 69.4, 68.7,
68.6, 68.0, 67.9, 67.8, 67.6, 67.6 (C-2 to C-5, C-20 to C-50, C-200 to C-500,
C-2G to C-5G), 61.9, 61.7 (C-6G), 50.3, 50.2, 50.1 (C-6, C-60, C-600),
24.1, 24.0, 23.7 (4 ꢁ CH2S), 21.3, 21.2, 21.0, 20.8 (CH3CO). Anal.
Calcd for C100H131N9O55S4: C, 47.88; H, 5.22; N, 6.70; S, 5.11. Found:
C, 47.69; H, 5.02; N, 6.99; S, 5.44. HRMS (ESI) m/z [M þ H]þ calcd
for C100H132N12O55S4 2507.6694, found 2507.6690.
Compound 21b: solvent system EtOAc/MeOH (98:2); yield 67%;
1
[R]20 þ3.0 (c 0.3, CHCl3); Rf 0.10 (EtOAc); H NMR (500 MHz,
D
DMSO-d6) δ 8.20, 8.07, 7.88 (3s, 1 H each, H-triazole), 6.22 (d, 1 H,
Compound 23b: solvent system EtOAc/MeOH (97:3); yield 48%;
[R]20D þ6.1 (c 1.0, CHCl3); Rf 0.53 (EtOAc/MeOH 1.95:1); 1H NMR
(500 MHz, DMSO-d6) δ 8.17, 8.11, 8.05, 7.83 (4 s, 4 H, H-triazole), 6.20
(d, 1 H, J1,2 = 9.2 Hz, H-1), 5.61 (t, 1 H, J2,3 = J3,4 = 9.2 Hz, H-3), 5.40
(m, 3 H, H-10, H-100, H-2), 5.32 (m, 6 H, H-30, H-300, 4 ꢁ H-4G), 5.17
J1,2 = 9.1 Hz, H-1), 5.59 (t, 1 H, J2,3 = J3,4 = 9.2 Hz, H-3), 5.47 (d, 1 H,
0
0
0
J1 ,2 = 3.4 Hz, H-1 ), 5.45 (t, 1 H, J1,2 = J2,3 = 9.3 Hz, H-2), 5.33 (m, 4 H,
H-30, 3 ꢁ H-4G), 5.18 (m, 3 H, 3 ꢁ H-3G), 5.01ꢀ4.89 (m, 8 H, H-20,
H-40, 3 ꢁ H-1G, 3 ꢁ H-2G), 4.72 (brd, 1 H, J5 ,6 a < 1 Hz, J6 a,6 b = 13.7
0
0
0
0
Hz, H-60a), 4.62 (dd, 1 H, J5 ,6 b = 6.5 Hz, J6 a,6 b = 13.8 Hz, H-60b),
4.56ꢀ4.41 (m, 9 H, 3 ꢁ CH2-triazole, H-6a, H-5, H-50), 4.25ꢀ4.15 (m,
4 H, H-6b, 3 ꢁ H-5G), 4.08ꢀ4.01 (m, 6 H, 3 ꢁ H-6aG, 3 ꢁ H-6bG),
3.92 (t, 1 H, J3,4 = J4,5 = 9.2 Hz, H-4), 3.62ꢀ3.50 (m, 18 H, 9 CH2O),
2.83, 2.72 (2 m, 6 H, 3 ꢁ CH2S), 2.13, 2.04, 2.03, 2.02, 2.00, 1.97, 1.93,
1.75 (8 s, 51 H, 17 ꢁ CH3CO); 13C NMR (125 MHz, DMSO-d6) δ
170.9, 170.8, 170.7, 170.5, 170.4, 170.3, 170.2, 170.1, 169.5 (CH3CO),
145.4, 144.9, 144.6 (C-4 triazole), 126.0, 125.9, 123.6 (C-5 triazole),
96.6 (C-10), 84.2 (C-1), 83.6 (3 ꢁ C-1G), 75.2 (C-4), 74.9 (C-3, C-5),
74.3 (3 ꢁ C-5G), 71.9 (3 ꢁ C-3G), 71.5 (C-2), 71.0, 70.3 (CH2O), 70.1,
69.9, 69.8, 69.7 (C-20, C-30, C-40, C-50), 68.5 (3 ꢁ C-4G), 68.2 (3 ꢁ
C-2G), 64.3, 64.2, 64.1 (3 ꢁ CH2-triazole), 62.5 (3 ꢁ C-6G), 50.7, 50.5
(C-6, C-60), 30.1 (3 ꢁ CH2S), 21.4, 21.2, 21.1 (CH3CO). Anal. Calcd
for C85H119N9O46S3: C, 48.64; H, 5.71; N, 6.01; S, 4.58. Found: C,
48.39; H, 5.79; N, 6.05; S, 4.45. HRMS (ESI) m/z [M þ Na]þ calcd for
C85H119N9NaO46S3 2120.6309, found 2120.6365.
(m, 4 H, 4 ꢁ H-3G), 5.01ꢀ4.89 (m, 9 H, H-200, 4 ꢁ H-1G, 4 ꢁ H-2G),
0
0
0
0
00
00 00
00 00
0
0
4.86 (t, 1 H, J3 ,4 = J4 ,5 = 9.7 Hz, H-4 ), 4.77 (dd, 1 H, J1 ,2 = 3.4 Hz,
J2 ,3 = 10.2 Hz, H-20), 4.71 (brd, 1 H, J5 ,6 a < 1 Hz, J6 a,6 b = 13.7 Hz,
0
0
0
0
0
0
H-60a), 4.62 (dd, 1 H, J5 ,6 b = 6.2 Hz, J6 a,6 b = 14.2 Hz, H-60b),
4.56ꢀ4.40 (m, 12 H, H-5, H-6a, H-500, H-600a, 4 ꢁ CH2Ar), 4.35ꢀ4.20
(m, 6 H, H-50, H-600b, 4 ꢁ H-5G), 4.12ꢀ4.00 (m, 9 H, H-6b, 4 ꢁ
H-6aG, 4 ꢁ H-6bG), 3.80 (m, 2 H, H-4, H-40), 3.63ꢀ3.50 (m, 24 H, 12
CH2O), 2.83, 2.74 (2 m, 8 H, 4 ꢁ CH2S), 2.13, 2.04, 2.03, 2.02, 2.00,
1.98, 1.96, 1.93, 1.75 (9s, 69 H, 23 ꢁ CH3CO); 13C NMR (125 MHz,
DMSO-d6) δ 170.9, 170.8, 170.7, 170.5, 170.4, 170.3, 170.2, 170.1, 169.5
(CH3CO), 145.3, 144.9, 144.5 (C-4 triazole), 126.3, 126.1, 123.7 (C-5
triazole), 96.5 (C-10, C-100), 84.2 (C-1), 83.6 (4 ꢁ C-1G), 76.0 (C-4),
75.2 (C-40), 75.0 (C-3, C-5), 74.4 (4 ꢁ C-5G), 71.9 (4 ꢁ C-3G), 71.7,
71.4, 71.1, 71.0, 70.6, 70.3, 70.2, 70.0, 69.9, 69.8, 69.7, 69.6, 69.5, 69.4 (C-
2, C-20, C-30, C-50, C-200 to C-500,CH2O), 68.5 (4 ꢁ C-4G), 68.1 (4 ꢁ
C-2G), 64.3, 64.1 (4 ꢁ CH2Ar), 62.5 (4 ꢁ C-6G), 50.7, 50.5, 50.4 (C-6,
C-60, C-600), 30.1 (4 ꢁ CH2S), 21.4, 21.3, 21.2, 21.1, 21.0, 20.7
(CH3CO). Anal. Calcd for C116H162N12O63S4: C, 48.70; H, 5.71; N,
5.88; S, 4.48. Found: C, 48.46; H, 5.89; N, 6.07; S, 4.65. HRMS (ESI) m/
z [M þ Na]þ calcd for C116H162N12NaO63S4 2881.8626, found
2881.8594.
0
0
0
0
Compound 21c: solvent system EtOAc/MeOH (96:4); yield 67%;
1
[R]20 þ0.9 (c 0.9, CHCl3); Rf 0.24 (EtOAc/MeOH 9:1); H NMR
D
(500 MHz, DMSO-d6) δ 8.19, 8.08, 7.88 (3s, 3 H, H-triazole), 6.22 (d, 1
H, J1,2 = 8.8 Hz, H-1), 5.59 (t, 1 H, J2,3 = J3,4 = 8.8 Hz, H-3), 5.47 (d, 1 H,
0
0
0
J1 ,2 = 3.4 Hz, H-1 ), 5.45 (t, 1 H, J1,2 = J2,3 = 8.9 Hz, H-2), 5.32 (m, 4 H,
H-30, 3 ꢁ H-4G), 5.17 (m, 3 H, 3 ꢁ H-3G), 5.02ꢀ4.88 (m, 8 H, H-20,
H-40, 3 ꢁ H-1G, 3 ꢁ H-2G), 4.62ꢀ4.40 (m, 11 H, 3 ꢁ CH2Ar, H-6a,
H-5, H-50, H-60a, H-60b), 4.25ꢀ4.15 (m, 4 H, H-6b, 3 ꢁ H-5G),
4.08ꢀ4.01 (m, 6 H, 3 ꢁ H-6aG, 3 ꢁ H-6bG), 3.91 (t, 1 H, J3,4 = J4,5 = 9.2
Hz, H-4), 3.65ꢀ3.47 (m, 54 H, 27 CH2O), 2.84, 2.72 (2 m, 3 H each,
3 ꢁ CH2S), 2.12, 2.04, 2.03, 2.02, 2.00, 1.97, 1.93, 1.74 (8 s, 51 H, 17 ꢁ
CH3CO); 13C NMR (125 MHz, DMSO-d6) δ 170.9, 170.8, 170.7,
170.5, 170.4, 170.3, 170.2, 170.0, 169.6 (CH3CO), 145.5 (C-4 triazole),
126.1, 123.7 (C-5 triazole), 96.6 (C-10), 84.2 (C-1), 83.6 (3 ꢁ C-1G),
75.3 (C-4), 74.9 (C-3, C-5), 74ꢀ3 (3 ꢁ C-5G), 71.9 (3 ꢁ C-3G), 71.5
(C-2), 71.1, 70.6, 70.5, 70.4 (CH2O), 70.1, 69.9, 69.8, 69.7 (C-20, C-30,
C-40, C-50), 68.5 (3 ꢁ C-4G), 68.1 (3 ꢁ C-2G), 64.3, 64.2, 64.1 (3 ꢁ
CH2Ar), 62.5 (3 ꢁ C-6G), 50.7, 50.6 (C-6, C-60), 30.1 (3 ꢁ CH2S),
21.4, 21.2, 21.1 (CH3CO). Anal. Calcd for C103H155N9O55S3: C, 49.57;
H, 6.26; N, 5.05; S, 3.85. Found: C, 49.42; H, 6.29; N, 5.02; S, 3.89.
HRMS (ESI) m/z [M þ Na]þ calcd for C103H155N9NaO55S3
2516.8668, found 2516.8620.
Compound 23c: solvent system EtOAc/MeOH (88:12); yield
1
54%; [R]20 þ5.4 (c 1.0, CHCl3); Rf 0.48 (EtOAc/MeOH 9:1); H
D
NMR (500 MHz, DMSO-d6) δ 8.17, 8.11, 8.06, 7.83 (4s, 4 H,
H-triazole), 6.20 (d, 1 H, J1,2 = 9.2 Hz, H-1), 5.61 (t, 1 H, J2,3 = J3,4
=
9.2 Hz, H-3), 5.41 (m, 3 H, H-10, H-100, H-2), 5.33 (m, 6 H, H-30, H-300,
4 ꢁ H-4G), 5.18 (dd, 4 H, J3G,4G = 3.6 Hz, J2G,3G = 9.8 Hz, 4 ꢁ H-3G),
4.99 (t, 4 H, J1G,2G = J2G,3G = 9.9 Hz, 4 ꢁ H-2G), 4.93 (m, 5 H, J1G,2G
=
00
10.0 Hz, H-200, 4 ꢁ H-1G), 4.86 (t, 1 H, J3 ,4 = J4 ,5 = 9.7 Hz, H-4 ),
00 00
00 00
0
0
0
0
0
0
0
4.77 (dd, 1 H, J1 ,2 = 3.5 Hz, J2 ,3 = 10.1 Hz, H-2 ), 4.71 (brd, 1 H, J5 ,6 a
<
=
1 Hz, J6 a,6 b = 13.7 Hz, H-60a), 4.62 (dd, 1 H, J5 ,6 b = 6.2 Hz, J6 a,6 b
0
0
0
0
0
0
14.2 Hz, H-60b), 4.57ꢀ4.38 (m, 12 H, H-5, H-6a, H-500, H-600a, 4 ꢁ
CH2Ar), 4.35ꢀ4.20 (m, 6 H, H-50, H-600b, 4 ꢁ H-5G), 4.08ꢀ4.00 (m, 9
H, H-6b, 4 ꢁ H-6aG, 4 ꢁ H-6bG), 3.79 (m, 2 H, H-4, H-40), 3.64ꢀ3.45
(m, 72 H, 36 CH2O), 2.83, 2.74 (2 m, 8 H, 4 ꢁ CH2S), 2.13, 2.04, 2.03,
2.02, 2.00, 1.98, 1.96, 1.93, 1.75 (9s, 69 H, 23 ꢁ CH3CO); 13C NMR
(125 MHz, CDCl3) δ 180.0, 170.9, 170.8, 170.7, 170.5, 170.4, 170.3,
170.2, 170.0, 169.6 (CH3CO), 145.4, 145.0, 144.6 (C-4 triazole), 126.3,
126.1, 126.0, 123.7 (C-5 triazole), 96.5 (C-10, C-100), 84.2 (C-1), 83.6
(3 ꢁ C-1G), 76.0 (C-4), 75.3 (C-40), 75.0 (C-3, C-5), 74.3 (4 ꢁ C-5G),
71.9 (4 ꢁ C-3G), 71.7, 71.4, 71.1, 71.0, 70.6, 70.3, 70.2, 70.0, 69.9, 69.8,
69.7, 69.6, 69.5, 69.4 (C-2, C-20, C-30, C-50, C-200 to C-500, CH2O), 68.5
(4 ꢁ C-4G), 68.1 (4 ꢁ C-2G), 64.3, 64.2, 64.1 (4 ꢁ CH2Ar), 62.6 (4 ꢁ
C-6G), 50.7, 50.5, 50.4 (C-6, C-60, C-600), 30.1 (4 ꢁ CH2S), 21.4, 21.3,
21.2, 21.1, 21.0; 20.7 (CH3CO). Anal. Calcd for C140H210N12O75S4: C,
49.61; H, 6.24; N, 4.96; S, 3.78. Found: C, 49.67; H, 6.54; N, 5.21; S, 3.87.
HRMS (ESI) m/z [M þ H]þ calcd for C140H211N12O75S4 3388.1948,
found 3388.1894.
Compound 23a: solvent system cyclohexane/EtOAc (92:8); yield
1
39%; [R]20 þ11.2 (c 0.7, CHCl3); Rf 0.50 (EtOAc); H NMR (500
D
MHz, DMSO-d6) δ 8.09, 8.00, 7.92, 7.77 (4s, 1 H each, H-triazole), 6.24
(d, 1 H, J1,2 = 9.1 Hz, H-1), 5.65 (t, 1 H, J2,3 = J3,4 = 9.2 Hz, H-3), 5.41
(m, 3 H, H-2, H-10, H-100), 5.33 (m, 6 H, H-30, H-300, 4 ꢁ H-4G), 5.19
(m, 4 H, 4 ꢁ H-3G), 5.02 (m, 4 H, 4 ꢁ H-2G), 4.92ꢀ4.35 (m, 16 H,
H-5, H-6a, H-6b, H-20, H-50, H-60a, H-60b, H-200, H-400, H-500, H-600a,
H-600b, 4 ꢁ H-1G), 4.23ꢀ4.15 (m, 4 H, 4 ꢁ H-5G), 4.11ꢀ3.86 (m, 17
H, H-4, 4 ꢁ CH2S, 4 ꢁ H-6aG, 4 ꢁ H-6bG), 2.14, 2.13, 2.12, 2.04, 2.02,
2.01, 2.00, 1.99, 1.98, 1.97, 1.96, 1.95, 1.94, 1.93, 1.92, 1.91, 1.90, 1.77
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ARTICLE
General Procedure for the O-Deacetylation. Compounds 11,
13aꢀc, 15aꢀc, 17aꢀc, 19aꢀc, 21aꢀc, and 23aꢀc (0.10 mmol) were
suspended in a mixture of MeOH/Et3N/H2O 4:1:5 (10 mL) and stirred
at room temperature. The solid was progressively dissolving and after 2 h
TLC (EtOAc or EtOAc:MeOH 19: 1) showed complete consumption
of the starting material. The solution was concentrated and the residue
was dissolved in water (1 mL) and passed through a column filled with
Dowex MR-3C mixed bed ion-exchange resin. The eluate was concen-
trated and further purified by filtration through an octadecyl C18
minicolumn. Evaporation of the solvent afforded the free product, which
showed a single spot by TLC (n-BuOH/EtOH/H2O 2.5:1:1) whose Rf
is indicated in each case.
H-6bG, 9 CH2O), 3.60 (dd, 1 H, J3G,4G = 3.3 Hz, J2G,3G = 9.4 Hz, H-3G),
5.12 (t, 1 H, J1G,2G = J2G,3G = 9.6 Hz, H-2G), 2.94, 2.88 (2 m, 2 H,
CH2S); 13C NMR (125 MHz, CDCl3) δ 144.7 (C-4 triazole), 124.8 (C-
5 triazole), 87.9 (C-1), 86.4 (C-1G), 79.4, 79.3 (C-5, C-5G), 76.4 (C-3),
74.4 (C-3G), 72.7 (C-2), 70.6, 70.0 (ꢁ6), 69.8, 69.5 (9 CH2O), 70.1 (C-
2G), 69.4, 69.3 (C-4, C-4G), 63.5 (CH2Ar), 61.5, 60,9 (C-6, C-6G), 29.6
(CH2S). Anal. Calcd for C25H45N3O15S: C, 45.52; H, 6.88; N, 6.37; S,
4.86. Found: C, 45.42; H, 6.85; N, 6.51; S, 4.70. HRMS (ESI) m/z [M þ
Na]þ calcd for C25H45N3NaO15S 682.2469, found 682.2479.
Compound 16a: yield 100%; mp 121 °C; [R]20 þ4.1 (c 0.4,
D
H2O); Rf 0.33; 1H NMR (500 MHz, D2O) δ 7.89 (s, 1 H, H-triazole),
4.66 (dd, 1 H, J5,6a = 2.5 Hz, J6a,6b = 14.6 Hz, H-6a), 4.58 (d, 1 H, J1,2
=
Compound 12: yield 87%; mp 50 °C; [R]20D ꢀ50.4 (c 0.6, H2O);
Rf 0.66; 1H NMR (500 MHz, D2O) δ 7.88 (s, 1 H, H-triazole), 4.86 (dd,
1 H, J = 6.5 Hz, J = 6.9 Hz, CHN), 4.28 (d, 1 H, J1,2 = 9.6 Hz, H-1), 4.00
(d, 1 H, Jgem = 14.6 Hz, CHS), 3.86 (dd, 1 H, J4,5 = 0.9 Hz, J3,4 = 3.5 Hz,
H-4), 3.88 (d, 1 H, Jgem = 14.6 Hz, CHS), 3.58 (m, 2 H, H-6a, H-6b),
3.53 (ddd, 1 H, J4,5 = 0.8 Hz, J5,6a = 5.4 Hz, J5,6b = 6.9 Hz, H-5), 3.50 (m,
2 H, H-2, H-3), 2.15, 1.86, 1.72, 1.64 (4 m, 4 H, 2 CH2cyclo); 13C NMR
(125 MHz, D2O) δ 144.8 (C-4 triazole), 122.8 (C-5 triazole), 85.2 (C-
1), 78.9 (C-5), 73.9, 69.4 (C-2, C-3), 68.7 (C-4), 62.4 (CHN), 60.9 (C-
1), 32.8 (ꢁ2) (C-2cyclo), 23.6 (ꢁ3) (C-3cyclo, CH2S). Anal. Calcd for
3.8 Hz, H-1), 4.47 (dd, 1 H, J5,6b = 7.9 Hz, J6a,6b = 14.6 Hz, H-6b), 4.21
(d, 1 H, J1G,2G = 9.7 Hz, H-1G), 3.97, 3.84 (2 d, 2 H, Jgem = 14.6 Hz,
CH2S), 3.81 (m, 1 H, H-4G), 3.80 (ddd, 1 H, J5,6a = 2.5 Hz, J5,6b = 7.9
Hz, J4,5 = 10.0 Hz, H-5), 3.59 (dd, 1 H, J5G,6aG = 7.7 Hz, J6aG,6bG = 11.7
Hz, H-6aG), 3.55 (dd, 1 H, J5G,6bG = 4.5 Hz, J6aG,6bG = 11.7 Hz, H-6bG),
3.51 (dd, 1 H, J3,4 = 9.2 Hz, J2,3 = 9.8 Hz, H-3), 3.50 (ddd, 1 H, J4G,5G
=
0.8 Hz, J5G,6bG = 4.5 Hz, J5,6aG = 7.7 Hz, H-5G), 3.43 (m, 2 H, H-2G,
H-3G), 3.37 (dd, 1 H, J1,2 = 3.8 Hz, J2,3 = 9.8 Hz, H-2), 3.07 (dd, 1 H,
J3,4 = 9.2 Hz, J4,5 = 10.0 Hz, H-4), 2.99 (s, 3 H, ꢀOCH3); 13C NMR (125
MHz, D2O) δ 145.0 (C-4 triazole), 125.3 (C-5 triazole), 99.1 (C-1),
84.9 (C-1G), 78.9 (C-5G), 73.9 (C-3G), 73.0 (C-3), 71.0 (C-2), 70.8
(C-4), 70.0 (C-5), 69.4 (C-2G), 68.7 (C-4G), 61.0 (C-6G), 54.8
C14H23N3O5S 0.5H2O: C, 47.44; H, 6.83; N, 11.86; S, 9.05. Found: C,
3
47.44; H, 6.76; N, 11.79; S, 8.86. HRMS (ESI) m/z [M þ Na]þ calcd for
C14H23N3NaO5S 368.1256, found 368.1251.
(ꢀOCH3), 50.9 (C-6), 23.3 (CH2S). Anal. Calcd for C16H27N3O10S
3
Compound 14a: yield 100%; mp 135ꢀ136 °C; [R]20D ꢀ22.8 (c
0.2, H2O); Rf 0.36;1H NMR (500 MHz, D2O) δ 8.10 (s, 1 H,
0.5H2O: C, 41.55; H, 6.10; N, 9.09; S, 6.93. Found: C, 41.14; H, 6.20; N,
8.81; S, 7.07. HRMS (ESI) m/z [M þ Na]þ calcd for C16H27N3NaO10S
476.1315, found 476.1316.
H-triazole), 5.58 (d, 1 H, J1,2 = 9.3 Hz, H-1), 4.26 (d, 1 H, J1G,2G
=
9.6 Hz, H-1G), 4.00, 3.88 (2 d, 2 H each, Jgem = 14.8 Hz, CH2S), 3.84 (t,
1 H, J1,2 = J2,3 = 9.3 Hz, H-2), 3.80 (dd, 1 H, J4G,5G = 0.8 Hz, J3G,4G= 2.1
Hz, H-4G), 3.76 (dd, 1 H, J5,6a = 1.8 Hz, J6a,6b = 12.2 Hz, H-6a), 3.63 (dd,
1 H, J5,6b = 5.3 Hz, J6a,6b = 12.2 Hz, H-6b), 3.59 (ddd, 1 H, J5,6a = 1.8 Hz,
J5,6b = 5.3 Hz, J4,5 = 9.4 Hz, H-5), 3.56 (t, 1 H, J2,3 = J3,4 = 9.3 Hz, H-3),
3.54 (dd, 1 H, J5G,6aG = 4.3 Hz, J6aG,6bG = 11.5 Hz, H-6aG), 3.50ꢀ3.45
(m, 4 H, H-3G, H-4, H-5G, H-6bG), 3.43 (t, 1 H, J1G,2G = J2G,3G = 9.5
Hz, H-2G); 13C NMR (125 MHz, D2O) δ 145.6 (C-4 triazole), 123.5
(C-5 triazole), 87.4 (C-1), 85.2 (C-1G), 78.8 (ꢁ2) (C-5, C-5G), 75.9
(C-3), 73.8 (C-3G), 72.2 (C-2), 69.4 (C-2G), 68.9 (C-4), 68.7 (C-4G),
Compound 16b: yield 89%; [R]20D þ63.7 (c 0.2, H2O); Rf 0.23;
1H NMR (500 MHz, D2O) δ 7.98 (s, 1 H, H-triazole), 4.70 (dd, 1 H,
J5,6a = 2.5 Hz, J6a,6b = 14.6 Hz, H-6a), 4.59 (d, 1 H, J1,2 = 3.8 Hz, H-1),
4.55 (s, 2 H, CH2-triazole), 4.49 (dd, 1 H, J5,6b = 8.0 Hz, J6a,6b = 14.6 Hz,
H-6b), 4.35 (d, 1 H, J1G,2G = 9.6 Hz, H-1G), 3.81 (m, 1 H, H-4G), 3.80
(ddd, 1 H, J5,6a = 2.5 Hz, J5,6b = 8.0 Hz, J4,5 = 10.1 Hz, H-5), 3.62ꢀ3.52
(m, 10 H, 3 ꢁ CH2O, H-3, H-5G, H-6aG, H-6bG), 3.47 (dd, 1 H,
J3G,4G = 3.3 Hz, J2G,3G = 9.4 Hz, H-3G), 3.40 (t, 1 H, J1G,2G = J2G,3G
=
9.3 Hz, H-2G), 3.38 (dd, 1 H, J1,2 = 3.8 Hz, J2,3 = 9.7 Hz, H-2), 3.09 (dd, 1
H, J3,4 = 9.1 Hz, J4,5 = 9.9 Hz, H-4), 2.98 (s, 3 H, ꢀOCH3), 2.83, 2.75 (2 m,
2 H, CH2S); 13C NMR (125 MHz, D2O) δ 144.1 (C-4 triazole), 126.3
(C-5 triazole), 99.2 (C-1), 85.9 (C-1G), 78.9 (C-5G), 73.9 (C-3G), 73.0
(C-3), 71.1 (C-2), 70.9 (C-4), 70.2, 69.3, 68.8 (3 ꢁ CH2O), 69.9 (C-5),
69.7 (C-2G), 68.8 (C-4G), 63.0 (CH2-triazole), 61.1 (C-6G), 54.8
60.9 (C-6G), 60.4 (C-6), 23.6 (CH2S). Anal. Calcd for C15H25N3O10S
3
H2O: C, 39.38; H, 5.95; N, 9.19; S, 7.00. Found: C, 39.50; H, 5.61; N,
8.96; S, 7.35. HRMS (ESI) m/z [M þ Na]þ calcd for C15H25N3NaO10S
462.1158, found 462.1173.
Compound 14b: yield 95%; mp 90 °C; [R]20D ꢀ7.4 (c 0.6, H2O);
(ꢀOCH3), 50.9 (C-6), 29.2 (CH2S). Anal. Calcd for C20H35N3O12S
3
Rf 0.29; 1H NMR (500 MHz, D2O) δ 8.20 (s, 1 H, H-triazole), 5.68 (d, 1
H2O: C, 42.93; H, 6.66; N, 7.51. Found: C, 42.60; H, 6.34; N, 7.70. HRMS
(ESI) m/z [M þ Na]þ calcd for C20H35N3NaO12S 564.1834, found
564.1850; m/z [M þ H]þ calcd for C20H36N3O12S 542.2014, found
542.2023.
0
0
H, J1,2 = 9.3 Hz, H-1), 4.64 (s, 2 H, CH2Ar), 4.42 (d, 1 H, J1 ,2 = 9.6 Hz,
H-1G), 3.92 (t, 1 H, J1,2 = J2,3 = 9.3 Hz, H-2), 3.87 (br d, 1 H, J3G,4G = 3.2
Hz, J4G,5G < 1.0 Hz, H-4G), 3.83 (dd, 1 H, J5,6a = 1.5 Hz, J6a,6b = 12.0 Hz,
H-6a), 3.72ꢀ3.61 (m, 10 H, 3 CH2O, H-3, H-6aG, H-6bG), 3.61ꢀ3.57
(m, 2 H, H-5, H-5G), 3.54 (t, 1 H, J3,4 ≈ J4,5 = 9.5 Hz, H-4), 3.52 (dd, 1
Compound 16c: yield 87%; [R]20D þ70.7 (c 0.2, H2O); Rf 0.16;
1H NMR (500 MHz, D2O) δ = 7.97 (s, 1 H, H-triazole), 4.70 (dd, 1 H,
J5,6a = 2.5 Hz, J6a,6b = 14.6 Hz, H-6a), 4.59 (d, 1 H, J1,2 = 3.8 Hz, H-1),
4.55 (s, 2 H, CH2-triazole), 4.49 (dd, 1 H, J5,6b = 8.1 Hz, J6a,6b = 14.6 Hz,
H-6b), 4.35 (d, 1 H, J1G,2G = 9.7 Hz, H-1G), 3.82 (dd, 1 H, J4G,5G = 0.6
Hz, J3G,4G = 3.4 Hz, H-4G), 3.80 (ddd, 1 H, J5,6a = 2.5 Hz, J5,6b = 8.1 Hz,
J4,5 = 10.1 Hz, H-5), 3.64ꢀ3.51 (m, 22 H, 9 ꢁ CH2O, H-3, H-5G,
H-6aG, H-6bG), 3.49 (dd, 1 H, J3G,4G = 3.4 Hz, J2G,3G = 9.3 Hz, H-3G),
3.40 (t, 1 H, J1G,2G = J2G,3G = 9.5 Hz, H-2G), 3.38 (dd, 1 H, J1,2 = 3.8 Hz,
J2,3 = 9.8 Hz, H-2), 3.09 (dd, 1 H, J3,4 = 9.1 Hz, J4,5 = 10.0 Hz, H-4), 2.98
(s, 3 H, ꢀOCH3), 2.84, 2.77 (2 m, 2 H, CH2S); 13C NMR (125 MHz,
D2O) δ 143.9 (C-4 triazole), 126.2 (C-5 triazole), 99.2 (C-1), 86.0 (C-
1G), 79.0 (C-5G), 73.9 (C-3G), 73.0 (C-3), 71.1 (C-2), 70.9 (C-4),
70.1, 69.9, 69.7, 69.6, 69.5, 69.4, 69.3, 69.2, 68.9, 68.8 (9 ꢁ CH2O, C-5,
C-2G, C-4G), 63.0 (CH2-triazole), 61.1 (C-6G), 54.8 (ꢀOCH3), 50.9 (C-
H, J2G,3G = 9.6 Hz, J3G,4G = 3.2 Hz, H-3G), 3.54 (t, 1 H, J1G,2G = J2G,3G
=
9.6 Hz, H-2G), 2.90, 2.80 (2 m, 2 H, J = 6.4 Hz, Jgem = 14.0 Hz, CH2S);
13C NMR (125 MHz, D2O) δ 144.2 (C-4 triazole), 124.4 (C-5 triazole),
87.4 (C-1), 85.9 (C-1G), 78.9 (ꢁ2) (C-5, C-5G), 75.9 (C-3), 73.9 (C-
3G), 72.2 (C-2), 70.2, 69.7, 69.3, 69.0, 68.9, 68.8 (C-2G, C-4, C-4G, 3
CH2O), 63.0 (CH2Ar), 61.0, 60.4 (C-6, C-6G), 29.1 (CH2S). Anal.
Calcd for C19H33N3O12S 0.5H2O: C, 42.53; H, 6.39; N, 7.83; S, 5.98.
3
Found: C, 42.57; H, 6.01; N, 7.97; S, 5.95. HRMS (ESI) m/z [M þ
Na]þ calcd for C19H33N3NaO12S 550.1677, found 550.1658.
Compound 14c. yield 90%; [R]20D ꢀ8.7 (c 0.2, D2O); Rf 0.25; 1H
NMR (500 MHz, CDCl3) δ 8.25 (s, 1 H, H-triazole), 5.73 (d, 1 H, J1,2
=
9.2 Hz, H-1), 4.70 (s, 2 H, CH2Ar), 4.46 (d, 1 H, J1G,2G = 9.7 Hz, H-1G),
3.97 (t, 1 H, J1,2 = J2,3 = 9.2 Hz, H-2), 3.93 (brd, 1 H, J4G,5G < 1 Hz,
J3G,4G = 3.3 Hz, H-4G), 3.87 (dd, 1 H, J5,6a = 5.0 Hz, J6a,6b = 11.8 Hz,
H-6a), 3.77ꢀ3.63 (m, 25 H, H-3, H-4, H-5, H-6b, H-5G, H-6aG,
6), 29.2 (CH2S). Anal. Calcd for C26H47N3O15S H2O: C, 45.14; H, 7.14;
3
N, 6.07. Found: C, 44.90; H, 6.98; N, 5.99. HRMS (ESI) m/z [M þ Na]þ
3074
dx.doi.org/10.1021/jo102421e |J. Org. Chem. 2011, 76, 3064–3077

The Journal of Organic Chemistry
ARTICLE
calcd for C26H47N3NaO15S 696.2620, found 696.2648; m/z [M þ H]þ
calcd for C26H48N3O15S 674.2801, found 674.2827.
4.52 (dd, 1 H, J5,6b = 8.2 Hz, J6a,6b = 14.6 Hz, H-6b), 4.34 (d, 1 H, H-1G),
4.09 (d, 1 H, Jgem = 14.5 Hz, CHS), 4.03 (m, 1 H, J5,6a = 2.2 Hz, J5,6b = 8.0
Hz, J4,5 = 10.2, H-5), 3.96 (m, 2 H, Jgem = 14.5 Hz, CHS, H-4G), 3.74 (t,
Compound 18a: yield 91%; mp 142ꢀ144 °C; [R]20D ꢀ46.7 (c 0.4,
1
1 H, J2,3 = J3,4 = 9.5 Hz, H-3), 3.70 (dd, 1 H, J5G,6aG = 7.7 Hz, J6aG,6bG
=
H2O); Rf 0.18; H NMR (500 MHz, D2O) δ 8.08, 7.80 (2 s, 2 H,
11.8 Hz, H-6aG), 3.67 (dd, 1 H, J5G,6bG = 4.5 Hz, J6aG,6bG = 11.8 Hz,
H-6bG), 3.60 (br dd, 1 H, J4G,5G < 1.0 Hz, J5G,6bG = 4.5 Hz, J5G,6aG = 7.7
Hz, H-5G), 3.55 (m, 2 H, H-2G, H-3G), 3.48 (dd, 1 H, J1,2 = 3.9 Hz, J2,3
= 9.9 Hz, H-2), 3.20 (t, 1 H, J3,4 = J4,5 = 9.6 Hz, H-4); 13C NMR (125
MHz, D2O) δ 145.4 (C-4 triazole), 125.6 (C-5 triazole), 93.7 (C-1),
85.4 (C-1G), 79.5 (C-5G), 74.3 (C-3G), 73.1 (C-3), 71.4 (C-4), 71.0
(C-2), 70.8 (C-5), 69.9 (C-2G), 69.2 (C-4G), 61.5 (C-6G), 51.3 (C-6),
23.7 (CH2S). Anal. Calcd for C30H48N6O19S2: C, 41.86; H, 5.62; N,
9.76; S, 7.45. Found: C, 41.97; H, 5.73; N, 8.91; S, 7.08. HRMS (ESI) m/
z [M þ Na]þ calcd for C30H48N6NaO19S2 883.2313, found 883.2336.
Compound 20b: yield 94%; [R]20D þ41.8 (c 0.3, D2O); Rf 0.17;
1H NMR (500 MHz, D2O) δ 8.02 (s, 1 H, H-triazole), 4.75 (dd, 1 H,
J5,6a = 1.6 Hz, J6a,6b = 14.6 Hz, H-6a), 4.65 (s, 2 H, CH2Ar), 4.58 (d, 1 H,
J1,2 = 4.2 Hz, H-1), 4.56 (dd, 1 H, J5,6b = 7.9 Hz, J6a,6b = 14.6 Hz, H-6b),
4.46 (d, 1 H, J1G,2G = 9.5, H-1G), 4.03 (m, 1 H, J5,6a = 1.7 Hz, J5,6b = 7.9
Hz, J4,5 = 9.6, H-5), 3.93 (d, 1 H, J4G,5G < 1 Hz, J3G,4G = 3.1 Hz, H-4G),
3.74 (t, 1 H, J2,3 = J3,4 = 9.4 Hz, H-3), 3.72ꢀ3.66 (m, 3 H, H-5G, H-6aG,
H-6bG), 3.66ꢀ3.62 (m, 6 H, 3 CH2O), 3.58 (dd, 1 H, J3G,4G = 3.1 Hz,
J2G,3G = 9.5 Hz, H-3G), 3.52 (t, 1 H, J1G,2G = J2G,3G = 9.5 Hz, H-2G),
H-triazole), 5.62 (d, 1 H, J1,2 = 9.2 Hz, H-1), 4.78 (dd, 1 H, J5,6a = 2.3 Hz,
J6a,6b = 15.0 Hz, H-6a), 4.60 (dd, 1 H, J5,6b = 7.1 Hz, J6a,6b = 15.0 Hz,
H-6b), 4.34, 4.12 (2 d, 2 H, J1G,2G = J1G ,2G = 9.6 Hz, H-1G, H-1G0),
4.06, 4.01, 3.95, 3.87 (4 d, 4 H, Jgem = 14.8 Hz, 2 ꢁ CH2S), 4.01 (ddd,
1 H, J5,6a = 2.3 Hz, J5,6b = 7.0 Hz, J4,5 = 9.7 Hz, H-5), 3.88, 3.82 (2 dd, 2 H,
0
0
0
0
0
0
0
J4G,5G = J4G ,5G = 0.8 Hz, J3G,4G = J3G ,4G = 3.2 Hz, H-4G, H-4G ), 3.81
(t, 1 H, J1,2 = J2,3 = 9.3 Hz, H-2), 3.64 (t, 1 H, J2,3 = J3,4 = 9.3 Hz, H-3),
3.62ꢀ3.51 (m, 6 H, H-5G, H-5G0, H6aG, H-6aG0, H-6bG, H-6bG0),
3.49 (dd, 1 H, J3G,4G = 3.0 Hz, J2G,3G = 9.5 Hz, H-3G*), 3.48 (t, 1 H, J3,4
=
0
0
0
0
J4,5 = 9.4 Hz, H-4), 3.42 (dd, 1 H, J3G ,4G = 3.3 Hz, J2G ,3G = 9.4 Hz,
H-3G0*), 3.33ꢀ3.29 (m, 2 H, H-2G, H-2G0); 13C NMR (125 MHz,
D2O) δ 145.6, 144.7 (C-4 triazole), 125.3, 123.2 (C-5 triazole), 87.3 (C-
1), 85.3, 84.5 (C-1G, C-1G0), 79.0, 79.9 (C-5G, C-5G0), 76.6 (C-5), 75.7
(C-3), 73.8 (ꢁ2) (C-3G, C-3G0), 72.2 (C-2), 70.0 (C-4), 69.4, 69.3 (C-
2G, C-2G0), 68.7 (ꢁ2) (C-4G, C-4G0), 61.0, 60.9 (C-6G, C-6G0), 50.7
(C-6), 23.6, 23.2 (2 ꢁ CH2S). Anal. Calcd for C24H38N6O14S2: C,
41.25; H, 5.48; N, 12.03; S, 9.18. Found: C, 40.93; H, 5.44; N, 11.93; S,
8.98. HRMS (ESI) m/z [M þ Na]þ calcd for C24H38N6NaO14S2
721.1785, found 721.1802.
Compound 18b: yield 93%; [R]20D ꢀ4.3 (c 0.6, D2O); Rf 0.15; 1H
NMR (500 MHz, D2O) δ 8.17, 7.92 (2 s, 2 H, H-triazole), 5.70 (d, 1 H,
J1,2 = 9.3 Hz, H-1), 4.85 (dd, 1 H, J5,6a = 2.4 Hz, J6a,6b = 14.8 Hz, H-6a),
4.70 (m, 1 H, J5,6b = 7.2 Hz, J6a,6b = 14.6 Hz, H-6b), 4.68, 4.61 (2 s, 4 H,
3.40 (dd, 1 H, J1,2 = 3.7 Hz, J2,3 = 9.8 Hz, H-2), 3.19 (t, 1 H, J3,4 = J4,5
=
C
9.5 Hz, H-4), 2.94, 2.86 (2 m, 2 H, J = 6.5 Hz, Jgem = 13.4 Hz, CH2S); 13
NMR (125 MHz, D2O) δ 144.3 (C-4 triazole), 126.5 (C-5 triazole),
93.8 (C-1), 86.4 (C-1G), 79.4 (C-5G), 74.4 (C-3G), 73.0 (C-3), 71.3
(C-4), 71.1 (C-2), 70.8 (C-5), 70.6, 69.8, 69.3 (CH2O), 70.1 (C-2G),
69.3 (C-4G), 63.4 (CH2Ar), 61.5 (C-6G), 51.3 (C-6), 29.6 (CH2S).
0
0
0
CH2Ar), 4.46, 4.45 (2 d, 2 H, J1G,2G = J1G ,2G = 9.7 Hz, H-1G, H-1G ),
4.07 (m, 1 H, J5,6a = 2.4 Hz, J5,6b = 7.2 Hz, J4,5 = 9.8 Hz, H-5), 3.93 (m,
2 H, H-4G, H-4G0), 3.91 (t, 1 H, J1,2 = J2,3 = 9.3 Hz, H-2), 3.73ꢀ3.56 (m,
21 H, H-3, H-3G, H-3G0, H-5G, H-5G0, H-6aG, H-6aG0, H-6bG,
Anal. Calcd for C38H64N6O23S2 H2O: C, 43.26; H, 6.31; N, 7.97; S,
3
6.08. Found: C, 43.53; H, 6.40; N, 7.51; S, 5.70. HRMS (ESI) m/z [M þ
H-6bG0, 6 CH2O), 3.51 (t, 2 H, J1G,2G = J1G ,2G = J2G,3G = J2G ,3G
=
0
0
0
0
Na]þ calcd for C38H64N6NaO23S2 1059.3362, found 1059.3342.
9.5 Hz, H-2G, H-2G0), 3.36 (t, 1 H, J3,4 = J4,5 = 9.5 Hz, H-4), 2.93, 2.84
(m, 4 H, 2 CH2S); 13C NMR (125 MHz, D2O) δ 144.3, 143.8 (C-4
triazole, 126.3, 124.2 (C-5 triazole), 87.2 (C-1), 85.9 (ꢁ2) (C-1G,
C-1G0), 78.9 (ꢁ2) (C-5G, C-5G0), 76.3 (C-5), 75.6 (C-3), 73.9 (ꢁ2)
(C-3G, C-3G0), 72.1 (C-2), 70.1 (ꢁ2), 70.0, 69.7 (ꢁ2), 69.3 (ꢁ2), 69.0,
68.8 (ꢁ2), 68.7 (C-2G, C-2G0, C-4, C-4G, C-4G0, 6 CH2O), 63.0, 62.9
(CH2Ar), 61.1 (ꢁ2) (C-6G, C-6G0), 50.6 (C-6), 29.1 (ꢁ2) (CH2S).
Anal. Calcd for C32H54N6O18S2: C, 43.93; H, 6.22; N, 9.61; S, 7.33.
Found: C, 43.64; H, 6.27; N, 9.87; S, 6.94. HRMS (ESI) m/z [M þ
Na]þ calcd for C32H54N6NaO18S2 897.2834, found 897.2829.
Compound 20c. yield 90%; [R]20 þ22.2 (c 0.7, D2O); Rf 0.20
D
1
(double development); H NMR (500 MHz, D2O) δ 8.01 (s, 1 H,
H-triazole), 4.76 (dd, 1 H, J5,6a = 1.4 Hz, J6a,6b = 14.4 Hz, H-6a), 4.65 (s,
2 H, CH2Ar), 4.58 (d, 1 H, J1,2 = 4.5 Hz, H-1), 4.55 (dd, 1 H, J5,6b = 7.8
Hz, J6a,6b = 14.5 Hz, H-6b), 4.45 (d, 2 H, J1G,2G = 9.7 Hz, H-1G), 4.04
(m, 1 H, J5,6a = 1.5 Hz, J5,6b = 8.1 Hz, J4,5 = 9.5 Hz, H-5), 3.93 (d, 1 H,
J4G,5G < 1 Hz, J3G,4G = 2.8 Hz, H-4G), 3.74 (t, 1 H, J2,3 = J3,4 = 9.5 Hz,
H-3), 3.72ꢀ3.66 (m, 6 H, H-5G, H-6aG, H-6bG), 3.66ꢀ3.62 (m, 18 H,
9 CH2O), 3.59 (dd, 1 H, J3G,4G = 2.9 Hz, J2G,3G = 9.4 Hz, H-3G), 3.52 (t,
1 H, J1G,2G = J2G,3G = 9.5 Hz, H-2G), 3.40 (dd, 1 H, J1,2 = 3.6 Hz, J2,3
=
Compound 18c: yield 92%; [R]20D ꢀ2.1 (c 1.0, D2O); Rf 0.11; 1H
NMR (500 MHz, D2O) δ 8.20, 7.94 (2 s, 1 H, H-triazole), 5.71 (d, 1 H,
J1,2 = 9.2 Hz, H-1), 4.86 (dd, 1 H, J5,6a = 2.4 Hz, J6a,6b = 14.8 Hz, H-6a),
9.8 Hz, H-2), 3.18 (t, 1 H, J3,4 = J4,5 = 9.6 Hz, H-4), 2.94, 2.87 (2 m, 2 H,
CH2S); 13C NMR (125 MHz, D2O) δ 144.3 (C-4 triazole), 126.5 (C-5
triazole), 93.8 (C-1), 86.4 (C-1G), 79.4 (C-5G), 74.4 (C-3G), 73.0 (C-
3), 71.4 (C-4), 71.2 (C-2), 70.8 (C-5), 70.6 (ꢁ2), 70.4 (ꢁ4), 70.2 (ꢁ2),
69.4 (CH2O), 70.1 (C-2G), 69.3 (C-4G), 63.5 (CH2Ar), 61.5 (C-6G),
4.70 (m, 1 H, H-6b), 4.63 (2s, 4 H, 2 ꢁ CH2Ar), 4.47 (d, 2 H, J1G,2G
=
0
0
0
J1G ,2G = 9.7 Hz, H-1G, H-1G ), 4.08 (m, 1 H, J5,6a = 2.0 Hz, J5,6b = 7.2
Hz, J4,5 = 9.2 Hz, H-5), 3.95 (m, 2 H, H-4G, H-4G0), 3.92 (t, 1 H, J1,2
=
51.3 (C-6), 29.6 (CH2S). Anal. Calcd for C50H88N6O29S2 H2O: C,
3
J2,3 = 9.3 Hz, H-2), 3.75ꢀ3.56 (m, 45 H, H-3, H-3G, H-3G0, H-5G,
45.52; H, 6.88; N, 6.37; S, 4.86. Found: C, 45.61; H, 7.07; N, 6.03; S,
4.74. HRMS (ESI) m/z [M þ Na]þ calcd for C50H88N6NaO29S2
1323.4935, found 1323.4979.
H-5G0, H-6aG, H-6aG0, H-6bG, H-6bG0, 18 CH2O), 3.50 (t, 2 H, J1G,2G
0
0
0
0
0
= J1G ,2G = J2G,3G = J2G ,3G = 9.5 Hz, H-2G, H-2G ), 3.38 (t, 1 H, J3,4
=
J4,5 = 9.4 Hz, H-4), 2.95, 2.89 (2 m, 4 H, CH2S); 13C NMR (50.28 MHz,
D2O) δ 144.8, 144.4 (C-4 triazole), 126.7, 124.7 (C-5 triazole), 87.8 (C-
1), 86.4 (ꢁ2) (C-1G, C-1G0), 79.4 (ꢁ2) (C-5G, C-5G0), 76.9 (C-5),
76.1 (C-3), 74.4 (ꢁ2) (C-3G, C-3G0), 72.6 (C-2), 70.7 (ꢁ3), 70.1
(ꢁ10), 69.9 (ꢁ2), 69.6 (ꢁ2), 69.3 (18 CH2O), 70.6 (C-4), 70.2 (C-2G,
C-2G0), 69.4 (C-4G, C-4G0), 63.5 (ꢁ2) (2 ꢁ CH2Ar), 61.6 (ꢁ2) (C-
6G, C-6G0), 50.2 (C-6), 29.6 (ꢁ2) (2 ꢁ CH2S). Anal. Calcd for
Compound 22a: yield 89%; [R]20 þ4.3 (c 0.5, D2O); Rf 0.21
D
(double development); 1H NMR (500 MHz, D2O) δ 8.06, 7.98, 7.73 (3s, 3
0
0
H, H-triazole), 5.58 (d, 1 H, J1,2 = 8.6 Hz, H-1), 5.36 (d, 1 H, J1 ,2 = 3.9 Hz,
H-10), 4.90 (br d, 1 H, J5 ,6 a < 1 Hz, J6 a,6 b = 14.1 Hz, H-60a), 4.52 (dd, 1 H,
0
0
0
0
J5 ,6 b = 8.7 Hz, J6 a, 6 b = 13.9 Hz, H-60b), 4.37, 4.30, 4.07 (3 d, 3 H, J1G,2G
=
0
0
0
0
9.1 Hz, 3 ꢁ H-1G), 4.15ꢀ3.84 (m, 9 H, H-2, H-3, H-5, H-6a, H-6b, H-50,
3 ꢁ H-4G), 3.79ꢀ3.40 (m, 25 H, H-4, H-20, H-30, H-40, 3 ꢁ H-2G, 3 ꢁ
H-3G, 3 ꢁ H-5G, 3 ꢁ H-6aG, 3 ꢁ H-6bG, 3 ꢁ CH2S); 13C NMR (125
MHz, D2O) δ 145.6, 144.8, 144.4 (C-4 triazole), 125.7, 125.5, 123.2 (C-5
triazole), 101.1 (C-10), 87.0 (C-1), 85.3, 85.2, 84.3 (3 ꢁ C-1G), 79.9, 79.1,
79.0, 76.0, 75.7, 75.6, 75.0, 73.9, 72.5, 72.4, 72.8, 71.7, 71.1, 71.0, 69.6, 69.5,
69.4, 68.7 (C-2 to C-5, C-20 to C-50, 3 ꢁ C-2G to C-5G), 61.1, 61.0, 60.7
(3 ꢁ C-6G), 51.5 (C-60), 50.2 (C-6), 23.6, 23.2, 23.0 (3 ꢁ CH2S). Anal.
C44H78N6O24S2 H2O: C, 45.67; H, 6.97; N, 7.26; S, 5.54. Found: C,
3
45.47; H, 7.01; N, 7.41; S, 5.72; HRMS (ESI) m/z [M þ Na]þ calcd for
C44H78N6NaO24S2 1161.4407, found 1161.4358.
Compound 20a: yield 84%; [R]20D þ10.9 (c 0.2, D2O); Rf 0.23;
1H NMR (500 MHz, D2O) δ 7.95 (s, 1 H, H-triazole), 4.74 (dd, 1 H,
J5,6a = 2.3 Hz, J6a,6b = 14.6 Hz, H-6a), 4.56 (d, 1 H, J1,2 = 3.9 Hz, H-1),
3075
dx.doi.org/10.1021/jo102421e |J. Org. Chem. 2011, 76, 3064–3077

The Journal of Organic Chemistry
ARTICLE
00 00
00
00
Calcd for C39H61N9O23S3: C, 41.82; H, 5.49; N, 11.25; S, 8.59. Found: C,
41.74; H, 5.72; N, 11.05; S, 8.23. HRMS (ESI) m/z [M þ H]þ calcd for
C39H62N9O23S3 1120.3121, found 1120.3118.
4.56 (dd, 1 H, J5 ,6 b = 9.3 Hz, J6 a, 6 b = 14.3 Hz, H-600b), 4.46 (m, 4 H,
4 ꢁ H-1G), 4.15ꢀ3.35 (m, 64 H, H-2 to H-5, H-6a, H-6b, H-20to H-50,
H-60a, H60b, H-200 to H-500, 4 ꢁ H-2G to H-5G, 4 ꢁ H-6aG, 4 ꢁ H-6bG,
12 CH2O), 2.90 (m, 8 H, 4 ꢁ CH2S); 13C NMR (125 MHz, D2O) δ
144.7, 144.4, 144.3, 144.2 (C-4 triazole), 126.4, 126.3, 124.3 (C-5 triazole),
101.0, 100.6 (C-10, C-10), 86.8 (C-1), 85.9 (4 ꢁ C-1G), 80.7, 80.3, 79.5,
76.3, 76.2, 75.2, 74.2, 73.0, 72.9, 72.8, 72.6, 71.9, 71.4, 71.3, 70.3, 69.9, 69.7,
69.6, 69.4, 69.3, 69.1, 68.8 (C-2 to C-5, C-20 to C-50, C-200 to C-500, 4 ꢁ
C-2G to C-5G, CH2O), 63.5, 63.4, 62.9 (4 ꢁ CH2Ar), 61.1 (C-6G), 50.9,
50.8, 50.3 (C-6, C-60, C-600), 29.2 (CH2S). Anal. Calcd for C70H116N12-
O40S4: C, 44.39; H, 6.17; N, 8.87; S, 6.77. Found: C, 44.56; H, 6.29; N,
9.03; S, 6.86. HRMS (ESI) m/z [M þ H]þ calcd for C70H117N12O40S4
1893.6362, found 1893.6318.
Compound 22b: yield 80%; [R]20 þ5.7 (c 0.8, D2O); Rf 0.19
D
1
1
(double development); H NMR (500 MHz, D2O) δ H NMR (500
MHz, D2O) δ 8.11, 8.07, 7.79 (3s, 3 H, H-triazole), 5.61 (d, 1 H, J1,2
=
0
0
0
0
0
8.7 Hz, H-1), 5.38 (d, 1 H, J1 ,2 = 3.9 Hz, H-1 ), 4.92 (br d, 1 H, J5 ,6 a < 1
Hz, J6 a,6 b = 13.8 Hz, H-60a), 4.60, 4.32 (3 s, 6 H, 3 ꢁ CH2Ar), 4.59 (dd,
0
0
1 H, J5 ,6 b = 8.7 Hz, J6 a, 6 b = 13.7 Hz, H-60b), 4.46 (d, 3 H, J1G,2G = 9.7
0
0
0
0
0
0
0
0
0
0
Hz, 3 ꢁ H-1G), 4.15 (ddd, 1 H, J5 ,6 a = 1.6 Hz, J5 ,6 b = 8.3 Hz, J4 ,5 = 9.9
Hz, H-50), 4.10ꢀ3.96 (m, 3 H, H-5, H-6a, H-6b), 3.94 (m, 5 H, H-2,
0
0
0
0
0
H-3, 3 ꢁ H-4G), 3.78 (t, 1 H, J2 ,3 = J3 ,4 = 9.5 Hz, H-3 ), 3.74ꢀ3.49 (m,
35 H, H-4, H-20, 3 ꢁ H-2G, 3 ꢁ H-3G, 3 ꢁ H-5G, 3 ꢁ H-6aG, 3 ꢁ
Compound 24c: yield 94%; [R]20 þ4.2 (c 0.3, D2O); Rf 0.13
0
0
0
0
0
H-6bG, 9 CH2O), 3.37 (t, 1 H, J3 ,4 = J4 ,5 = 9.2 Hz, H-4 ), 2.93, 2.82 (2
m, 6 H, CH2S); 13C NMR (125 MHz, D2O) δ 144.8, 144.3, 144.2 (C-4
triazole), 126.9, 126.7, 124.7 (C-5 triazole), 101.4 (C-10), 87.3 (C-1),
86.4 (C-1G), 80.2 (C-4), 79.4 (C-5G), 76.2 (C-3), 75.1 (C-5), 74.4 (C-
3G), 72.9 (C-30), 72.5 (C-50), 72.1 (C-20), 72.0 (C-2), 71.4 (C-40), 70.6,
70.5, 70.4, 70.1, 69.8, 69.7, 69.5, 69.3, 69.1 (C-2G, C-4G, 9 CH2O), 63.5,
63.4, 63.3 (3 ꢁ CH2Ar), 61.5 (C-6G), 51.9 (C-60), 50.4 (C-6), 29.6
(CH2S). Anal. Calcd for C51H85N9O29S3: C, 44.24; H, 6.19; N, 9.11; S,
6.95. Found: C, 44.05; H, 6.38; N, 9.35; S, 7.12. HRMS (ESI) m/z [M þ
H]þ calcd for C51H86N9O29S3 1384.4694, found 1384.4670.
D
1
(triple development); H NMR (500 MHz, D2O) δ 8.10, 8.04, 7.95,
7.76 (4s, 4 H, H-triazole), 5.59 (d, 1 H, J1,2 = 8.0 Hz, H-1), 5.36, 5.30
0
00
0
0
00 00
00 00
(2 d, 2 H, J1 ,2 = J1 ,2 = 3.4 Hz, H-1 , H-1 ), 4.93 (br d, 1 H, J5 ,6 a < 1
Hz, J6 a,6 b = 14.1 Hz, H-600a), 4.66, 4.64, 4.35, 4.29 (4 d, 8 H, 4 ꢁ
00
00
CH2Ar), 4.56 (dd, 1 H, J5 ,6 b = 9.2 Hz, J6 a, 6 b = 14.4 Hz, H-600b), 4.46
(d, 4 H, J1G,2G = 9.6 Hz, 4 ꢁ H-1G), 4.15ꢀ3.35 (m, 112 H, H-2 to H-5,
H-6a, H-6b, H-20to H-50, H-60a, H60b, H-200 to H-500, 4 ꢁ H-2G to
H-5G, 4 ꢁ H-6aG, 4 ꢁ H-6bG, 36 CH2O), 2.90 (m, 8 H, 4 ꢁ CH2S);
13C NMR (125 MHz, D2O) δ 144.7, 144.4, 144.3, 144.2 (C-4 triazole),
126.8, 126.7, 124.7 (C-5 triazole), 101.4, 101.0 (C-10, C-100), 87.3 (C-1),
86.4 (4 ꢁ C-1G), 81.0, 80.2, 79.4, 76.2, 76.1, 75.0, 74.4, 73.1, 73.0, 72.9,
72.8, 71.9, 71.8, 71.4, 71.3, 70.4, 70.3, 70.2, 69.9, 69.7, 69.6, 69.5, 69.4,
69.3, 69.2, 69.1, 68.8 (C-2 to C-5, C-20 to C-50, C-200 to C-500, 4 ꢁ C-2G
to C-5G, CH2O), 63.5, 63.4, 63.3, 63.2 (4 ꢁ CH2-triazole), 61.5 (C-6G),
51.9, 51.2, 50.3 (C-6, C-60, C-600), 29.6 (CH2S). Anal. Calcd for
C94H164N12O52S4: C, 46.60; H, 6.82; N, 6.94; S, 5.29. Found: C,
46.89; H, 7.11; N, 7.09; S, 5.45. HRMS (ESI) m/z [M þ H]þ calcd
for C94H165N12O52S4 2421.9519, found 2421.9522.
00 00
00
00
Compound 22c: yield 96%; [R]20 þ10.7 (c 0.4, D2O); Rf 0.12
D
(double development); 1H NMR (500 MHz, D2O) δ 8.10, 8.05, 7.78
0
0
(3s, 3 H, H-triazole), 5.60 (d, 1 H, J1,2 = 8.8 Hz, H-1), 5.38 (d, 1 H, J1 ,2
=
3.4 Hz, H-10), 4.91 (br d, 1 H, J5,6 a < 1 Hz, J6 a,6 b = 14.4 Hz, H-60a), 4.60
(m, 5 H, H-60b, 2 ꢁ CH2Ar), 4.44 (d, 3 H, J1G,2G = 9.6 Hz, 3 ꢁ H-1G),
4.32 (m, 2 H, CH2Ar), 4.15ꢀ4.00 (m, 4 H, H-5, H-6a, H-6b, H-50), 3.94
(m, 5 H, H-2, H-3, 3 ꢁ H-4G), 3.73ꢀ3.49 (m, 72 H, H-4, H-20, H-30,
0
0
0
3 ꢁ H-2G, 3 ꢁ H-3G, 3 ꢁ H-5G, 3 ꢁ H-6aG, 3 ꢁ H-6bG, 27 CH2O),
0
0
0
0
0
3.34 (t, 1 H, J3 ,4 = J4 ,5 = 9.2 Hz, H-4 ), 2.93, 2.85 (2 m, 6 H, 3 ꢁ CH2S);
13C NMR (125 MHz, D2O) δ 144.8, 144.3, 144.2 (C-4 triazole), 126.8,
126.6, 124.6 (C-5 triazole), 101.4 (C-10), 87.4 (C-1), 86.4 (C-1G), 80.2
(C-4), 79.4 (C-5G), 76.3 (C-3), 75.2 (C-5), 74.4 (C-3G), 72.9 (C-30),
72.4 (C-50), 72.1 (C-20), 72.0 (C-2), 71.4 (C-40), 70.6, 70.1, 69.8, 69.7,
69.5, 69.3, 69.1 (C-2G, C-4G, CH2O), 63.5, 63.4, 63.3 (3 ꢁ CH2-
triazole), 61.5 (C-6G), 51.9 (C-60), 50.4 (C-6), 29.6 (3 ꢁ CH2S). Anal.
Calcd for C69H121N9O38S3: C, 46.53; H, 6.85; N, 7.08; S, 5.40. Found:
C, 46.76; H, 7.09; N, 7.34; S, 5.69. HRMS (ESI) m/z [M þ H]þ calcd
for C69H122N9O38S3 1780.7044, found 1780.7053.
Enzymatic Assays:
Stability of the Glycoclusters toward the β-Galactosidase
A solution of the glycoclusters (1 mM) in sodium phosphate buffer
(100 mM, pH 7.3, MgCl2 1.2 mM, 2-mercaptoethanol 100 mM) was
incubated with the β-galactosidase from E. coli (0.6 U/mL) at 37 °C.
Aliquots of the solution were taken after 8, 16, and 24 h, evaporated, and
examined by TLC and NMR.
Inhibition of β-Galactosidase. The inhibitory activity of com-
pounds 12, 14aꢀc, 16aꢀc, 18aꢀc, 20aꢀc, 22aꢀc, and 24aꢀc toward
E. coli β-galactosidase (grade VIII, Sigma, EC 3.2.1.23, 117 U/mg) was
determined under standard conditions.16,17 The enzyme (0.3 U; 1U = 1
enzyme unit hydrolyses 1 μmol of o-nitrophenyl galactopyranoside per
minute) was incubated with o-nitrophenyl-β-D-galactopyranoside (con-
centration range: 0.2 to 5.0 mM) in sodium phosphate buffer (100 mM,
pH 7.3, MgCl2 1.2 mM, 2-mercaptoethanol 100 mM) in the absence or
presence of the free glycoclusters (12, 14aꢀc, 16aꢀc, 18aꢀc, 20aꢀc,
22aꢀc, and 24aꢀc; concentration range: 0.6 to 4.0 mM); the final
volume was 0.50 mL. After incubation for 10 min at 37 °C, the reaction
was quenched by addition of sodium borate buffer 0.2 M (4.0 mL, pH
10.0). The release of o-nitrophenol was measured by visible absorption
spectroscopy at 410 nm. The Ki and Km values and the inhibition type
were determined from LineweaverꢀBurk plots. The errors were esti-
mated by the linear regression standard method.
Compound 24a: yield 89%; [R]20 þ7.6 (c 0.2, D2O); Rf 0.12
D
(double development); 1H NMR (500 MHz, D2O) δ 8.06, 7.98, 7.91,
7.73 (4s, 4 H, H-triazole), 5.58 (d, 1 H0,0J1,2 = 8.6 Hz, H-1), 5.31 (2d, 1 H
0
0
0
00 00
00 00
each, J1 ,2 = J1 ,2 = 3.9 Hz, H-1 , H-1 ), 4.90 (br d, 1 H, J5 ,6 a < 1 Hz,
J6 a,6 b = 14.1 Hz, H-600a), 4.52 (dd, 1 H, J5 ,6 b = 8.7 Hz, J6 a, 6 b = 13.9
Hz, H-600b), 4.42, 4.37, 4.30, 4.07 (4 d, 4 H, J1G,2G = 9.1 Hz, 4 ꢁ H-1G),
4.19ꢀ3.33 (m, 48 H, H-2 to H-5, H-6a, H-6b, H-20 to H-50, H-60a,
H-60b, H-200 to H-500, 4 ꢁ H-2G to H-5G, 4 ꢁ H-6aG, 4 ꢁ H-6bG, 4 ꢁ
CH2S); 13C NMR (125 MHz, D2O) δ 146.1, 146.0, 145.1 (C-4
triazole), 126.3, 126.1, 123.6 (C-5 triazole), 101.2, 101.1 (C-10, C-100),
87.4 (C-1), 85.7, 85.6, 85.5, 84.8 (4 ꢁ C-1G), 81.3, 80.3, 79.5, 79.4, 79.3,
76.1, 75.3, 74.3, 72.9, 72.2, 71.5, 70.0, 69.9, 69.8, 69.7, 69.6, 69.4, 69.3,
69.2, 69.1 (C-2 to C-5, C-20 to C-50, C-200 to C-500, 4 ꢁ C-2G to C-5G),
61.7, 61.6, 61.5, 61.4 (4 ꢁ C-6G), 51.9, 51.3, 50.5 (C-6, C-60, C-600),
24.0, 23.6, 23.5, 23.4 (4 ꢁ CH2S). Anal. Calcd for C54H84N12O32S4: C,
42.07; H, 5.49; N, 10.90; S, 8.32. Found: C, 42.34; H, 5.66; N, 11.12; S,
8.01. HRMS (ESI) m/z [M þ Na]þ calcd for C54H84N12NaO32S4
1563.4095, found 1563.4099.
00
00
00
0
00
00
’ ASSOCIATED CONTENT
Supporting Information. 1H and 13C NMR spectra for
S
b
Compound 24b: yield 81%; [R]20 þ3.2 (c 0.3, D2O); Rf 0.18
compounds 2bꢀc, 3aꢀc, 11, 12, 13aꢀc, 14aꢀc, 15aꢀc, 16aꢀc,
17aꢀc, 18aꢀc, 19aꢀc, 20aꢀc, 21aꢀc, 22aꢀc, 23aꢀc, and
24aꢀc and COSY and HSQC for compounds 13b, 14b, 15b,
16b, 17a, 18a, 19a, 20a, 21b, 22b, 23c, and 24c. This material is
available free of charge via the Internet at http://pubs.acs.org.
D
(triple development); 1H NMR (500 MHz, D2O) δ 8.09, 8.05, 7.96, 7.76
(4s, 4 H, H-triazole), 5.60 (d, 1 H, J1,2 = 8.0 Hz, H-1), 5.38, 5.33 (2d, 1 H
each, J1 ,2 = J1 ,2 = 3.3 Hz, H-10, H-100), 4.93 (br d, 1 H, J5 ,6 a < 1 Hz,
0
0
00 00
00 00
J6 a,6 b = 14.1 Hz, H-600a), 4.66, 4.64, 4.35, 4.29 (4 d, 8 H, 4 ꢁ CH2Ar),
00
00
3076
dx.doi.org/10.1021/jo102421e |J. Org. Chem. 2011, 76, 3064–3077

The Journal of Organic Chemistry
ARTICLE
’ AUTHOR INFORMATION
(13) (a) Jacobson, R. H.; Zhang, X. J.; DuBose, R. F.; Matthews,
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Corresponding Author
*M.L.U.: phone/fax: þ541145763346; e-mail: mluhrig@
qo.fcen.uba.ar; J.K.: phone: þ33322827567; fax: þ33322827568;
e-mail: jose.kovensky@u-picardie.fr.
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’ ACKNOWLEDGMENT
Support of this work by the University of Buenos Aires
(project X227), the National Research Council of Argentina
(CONICET, Project PIP 0064), the Centre National de la
Recherche Scientifique, the Ministꢁere Dꢀelꢀeguꢀe ꢁa l’Enseignement
Supꢀerieur etꢁa la Recherche, and the Conseil Rꢀegional de Picardie
is gratefully acknowledged. M.L.U. and O.V. and are Research
Members from CONICET, A.J.C. is a fellow from CONICET. A.
ꢀ
J.C. thanks the Programme Egide des Bourses d’Excellence Eiffel.
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