Click multivalent homogeneous neoglycoconjugates - synthesis and evaluation of their binding affinities

Article abstract of DOI:10.1002/ejoc.200801170

Mannose (a-Man)-containing neoglycoconjugates possessing aliphatic-, aromatic-, and carbohydrate-centered architectures and differing in structural characteristics such as valency, topology, and nature of the linker have been synthesized using click chemi

Full text of DOI:10.1002/ejoc.200801170

FULL PAPER
DOI: 10.1002/ejoc.200801170
Click Multivalent Homogeneous Neoglycoconjugates – Synthesis and
Evaluation of Their Binding Affinities
Francisco Perez-Balderas,[a] Julia Morales-Sanfrutos,[a] Fernando Hernandez-Mateo,[a]
Joaquín Isac-García,[a] and Francisco Santoyo-Gonzalez*[a]
Dedicated to Professor Hans Helmut Baer
Keywords: Carbohydrates / Glycoconjugates / Multivalency / Cycloaddition / Click chemistry / Ligand design
Mannose (α-Man)-containing neoglycoconjugates pos- multivalent neoglycoconjugates. The binding behaviour of
sessing aliphatic-, aromatic-, and carbohydrate-centered
architectures and differing in structural characteristics such
as valency, topology, and nature of the linker have been syn-
thesized using click chemistry that shows its value as an ef-
ficient and versatile methodology for accessing tailor-made
these glycomimics toward Concanavalin A (Con A) has been
evaluated to determine the influence of the structural param-
eters.
(© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim,
Germany, 2009)
valent carbohydrate systems, the CuI-catalyzed 1,3-dipolar
cycloaddition of azides and alkynes (CuAAC),[12] nowadays
the best example under the “click-chemistry” concept,[13]
has reached a relevant place in recent times as a powerful
ligation method for glycoconjugation.[14] Owing to its reli-
ability, efficiency, robustness, chemoselectivity, and toler-
ance to a variety of reaction conditions, this highly modular
coupling reaction is especially useful for the effective con-
struction of complex glycosylated structures. In addition,
carbohydrates can easily be outfitted with an azido group,
either on the anomeric center or by a spacer and, similarly,
the introduction of an alkyne group is straightforward. In
common with other ligating methodologies, monofunction-
alized sugars are normally grafted onto polyfunctionalized
scaffolds or solid supports giving rise to molecular glyco-
systems containing multiple 1,2,3-triazole rings as tethering
groups.
For the synthesis of multivalent glycomimetics by means
of click-chemistry reactions, diverse polyalkyne- as well as
polyazide-functionalized scaffolds have been reacted with
complementary monoazide or monoalkyne sugar deriva-
tives.[14] The number of functions expressed in the scaffolds
is the variable that allows access to neoglycoconjugates with
different sugar densities and topologies. The scaffold struc-
ture and the nature of the spacer between the core and the
clicking groups are normally the factors that provide vari-
ability to reach structures with diverse cluster presentation
and lengths between the sugar moiety and the core. These
variables are essential to gaining fundamental insights into
the spatial factors regulating the binding of carbohydrates
to maximize the individual carbohydrate-receptor interac-
tions.
Introduction
The efficient construction of molecular systems bearing
multiple carbohydrate appendages has become a necessity
in the Glycobiology and Glycomic fields. The main reason
for this requirement is based on the prevalence of the multi-
valent principle[1] in the ligand-receptor interactions in
which carbohydrates are involved.[2] This effort to emulate
Nature has led to the development of multivalent glyco-
mimics[3] with different architectures (glycocluster,[4] glyco-
dendrimers,[4,5] glycocyclodextrins,[6,7] glycocalixarenes,[7]
glycopolymers,[8] and liposomes[9]) and carbohydrate densi-
ties to endow them with binding power greater than that of
their monovalent analogues. These glycomimics have at-
tracted attention because of their great potential not only to
gain insight into the molecular recognition events in which
carbohydrates participate but also for their capabilities in
biotechnology, pharmaceutical and medical applications.[10]
A second reason has been the demand of reliable techniques
for the immobilization of carbohydrates in the preparation
of carbohydrate-based surfaces. This driving force has led
to the development of a variety of glycofunctionalized solid
supports (microarrays, microbeads, biosensor chips, and
self-assembled monolayers) as valuable tools with impor-
tant applications in the “omic” sciences.[11]
Among the different reported methodologies for the co-
valent assembling of the constitutive modules of these poly-
[a] Departamento de Química Orgánica, Facultad de Ciencias, In-
stituto de Biotecnología
Universidad de Granada, 18071 Granada, Spain
Fax: +34-958-243-186
E-mail: fsantoyo@ugr.es
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In continuing our standing interest in the implementa-
tion of the click-chemistry methodology[15] for the prepara-
tion of multivalent neoglycoconjugates[16,17] and carbo-
hydrate-based materials,[18] the synthesis of a series of α-
Man-containing, homogeneous, multivalent glycoconju-
gates differing in the nature of the core, valency, topology,
and nature of the sugar-core linker was envisaged in order
to broaden the structural diversity of the aforementioned
glycoconjugates. We sought to gain fundamental insights
into the influence of those structural factors when these
clicked multivalent systems are used as glycomimics toward
natural lectins. This study is complemented by the synthesis
of heterogeneous multivalent systems by a modular click
strategy and the evaluation of their binding affinities as re-
ported in the next article of this issue.[19]
Figure 1. Click strategies for the synthesis of homogeneous multi-
valent neoglycoconjugates.
the thioglycoside 1b to assess the potential influence of the
sulfur on the binding affinity and to take advantage of its
higher hydrolytic stability relative to the corresponding O-
glycoside analogs.
Results and Discussion
Chemistry
α-Man was the sugar of choice considering the wide use
of Concanavalin A (Con A), the tetrameric plant lectin
from Canavalia ensiformis, as a suitable model lectin that
specifically recognizes α-Man residues.
We used two different chemical strategies for accessing
the desired multivalent neoglycoconjugates by means of the
CuAAC reaction as the ligating tool for the assembly of the
different constitutive building blocks: Strategy A, based in
the use of a polyazide scaffold, and Strategy B, based on
the use of a polyalkyne scaffold (Figure 1).
Figure 2. Monofunctionalized clickable alkynyl and azido α-Man
derivatives.
We used a variety of aromatic-centered polyazides (3B–
In both cases, we treated the functionalized scaffolds 3I, Figure 3) differing in their valency (from two to six),
with adequate, complementary, monofunctionalized, and substitution pattern as scaffolds for strategy A. The
clickable, α-Man derivatives (Figure 2). Thus, we selected azido group was linked to the aromatic ring through a
the alkynyl glycoside 1a, the alkynyl thioglycoside 1b, the methyl group except in the case of 3E where the linker was
azido α-Man 2c, and the azido ethyl α-Man 2d α-Man de- an ethoxy chain. In addition, we also used 1,2-diazido-
rivatives considering their easy preparation. We also chose ethane (3A) as a simple, homobifunctional, α-Man-contain-
Figure 3. Polyazide (3A–3I) and polyalkyne scaffolds (4J–4M).
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Click Multivalent Homogeneous Neoglycoconjugates
ing neoglycoconjugate to be used as a model compound
We carried out the CuAAC reactions using the clickable,
together with methyl α-Man in the biological assays. For α-Man, peracetylated derivatives to facilitate solubility in
strategy B, we selected pentaerythritol, methyl α--gluco- organic media, where the cycloadditions were performed
pyranoside, -glucose, and -mannitol as polyhydroxylated (Table 1 and Table 2). We conducted these cycloaddition re-
compounds and alkylated them to access the corresponding actions using the Cu catalyst (EtO)3P·CuI (10 mol-%) in
perpropargylated derivatives 4J–4M (Figure 3) to be used toluene under microwave irradiation or at reflux, conditions
as polyalkyne scaffolds with valencies ranging from four to that we found optimal previously.[16–18] We initially per-
six.
formed the reactions in the absence of any base. However,
in some cases (Method B, Table 1) we found that the pres-
ence of DIPEA favored the ligation of the complementary
clickable counterparts. We note that the reaction of the
hexamethylazide benzene 3I with the alkynyl thioglycoside
1b under these conditions failed for the complete grafting
of all the azidomethyl arms in spite of extended reaction
times. In this case, we obtained the hexavalent neoglycocon-
jugate 5Ib in DMF by heating the reaction mixture at 80 °C
in the presence of DIPEA.
Table 1. Synthesis of multivalent, homogeneous, neoglycoconju-
gates 5[a] from polyazide scaffolds and monoalkyne sugars.
Table 2. Synthesis of multivalent homogeneous neoglycoconjugates
6[a] from polyalkyne scaffolds and monoazide sugars.
[a] Compound (reaction time [h], % yield).
In the majority of the cases, the reaction were finished in
very short reaction times, particularly when we used micro-
wave irradiation, and we isolated the clicked, multivalent,
homogeneous neoglycoconjugates in high yields. We per-
[a] Reaction conditions: A: (EtO)3P·CuI, toluene, microwave irradi-
ation; B: (EtO)3P·CuI, toluene, DIPEA, microwave irradiation; C:
(EtO)3P·CuI, DIPEA, DMF, 80 °C. [b] Compound (reaction condi-
tions, reaction time [min], % yield).
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formed de-O-protection of the peracetylated derivatives ob- their relative binding inhibitory properties against perox-
tained using NaOMe or Et3N in MeOH to obtain the cor- idase-labeled Con A (see Table 3). All new compounds were
responding hydroxylated 7 and 8, which we evaluated for adequately characterized by spectroscopic techniques.
Table 3. Synthesis and binding affinities of multivalent homogeneous hydroxylated neoglycoconjugates 7 and 8.
[a] Compound (Method, % yield). Method A: Et3N, MeOH, reflux, 2–3 h; Method B: NaOMe, MeOH, room temp., 2–3 h.
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Click Multivalent Homogeneous Neoglycoconjugates
Lectin Affinity Evaluation
aromatic substituent at the anomeric position of glycoside
derivatives upon the binding to Con A and related lec-
The affinities of the hydroxylated, homogeneous, multi- tins.[21]
valent neoglycoconjugates 7 and 8 toward Con A were
An overall analysis of the relative potency per α-Man
evaluated by the ELISA-type protocol, ELLA (Table 3). unit values allows for the determination of the influence of
This experiment measures the capacity of a soluble ligand the valency on the binding properties of the neoglycoconju-
to inhibit lectin binding to a polymeric ligand that is used gates. Thus, we observed a positive effect when the valency
as a coating material on the microtiter well. In the present increased from two to three. However, we observed no sig-
case, we carried out competitive experiments using horse- nificant increases in affinity on going from trivalent to tet-
radish peroxidase-labeled Con A (HRP-Con A) as the lectin ravalent systems, and we even noted a decrease in penta-
and yeast mannan as the microplate-fixed ligand in tripli- and hexavalent systems.
cate. We included methyl α-Man in the tests as a reference
Concerning the influence of the anomeric atom, we note
compound. We considered up to six different concentra- that in all the aromatic-centered systems, we observed bet-
tions of each sample, and the percentage of the inhibition ter binding for the thioglycoside-containing systems in sys-
of HRP-Con A-yeast mannan association was determined tems of similar substitution than for their O-glycoside-con-
spectrophotometrically. We determined the IC50 values, de- taining counterparts, except for the trivalent electronic-rich
fined as the concentration of synthetic compound required systems (7Ee, 7Ef, 7Ge, and 7Gf), where the tendency was
to achieve 50% inhibition of this association, from the cor- opposite, as mentioned above.
responding inhibition curves. We assumed the IC50 values
Finally, we note that in all the compounds obtained from
to be inversely proportional to the corresponding free en- polyalkyne scaffolds by the strategy B (Table 3, Entries 21–
ergy of binding. The binding affinities of the references and 26) the presence of an ethoxy linker at the anomeric posi-
the homo neoglycoconjugates for Con A are summarized in tion had a beneficial effect on the inhibitory properties of
Table 3. The valency-corrected relative binding potencies these multivalent neoglycoconjugates, as we observed
are expressed per mol of α-Man residue relative to the higher relative affinities for 8Jh, 8Kh, 8Lh, and 8Mh than
monovalent methyl α-Man.
for 8Jg, 8Kg, 8Lg, and 8Mg, where the anomeric carbon is
An analysis of the binding affinity values found for these directly grafted to the triazole ring.
novel homo neoglycoconjugates lead to the following con-
clusions. First, in the case of divalent systems (Table 3, En-
tries 1–8), we observed that aliphatic-centered systems (7Ae
and 7Af) show lower relative affinities than do systems con-
Conclusions
In summary, we performed the synthesis of structurally
structed using an aromatic scaffold (7Be, 7Bf, 7Ce, 7Cf,
7De, and 7Df). In these aromatic-centered glycoconjugates,
we also observed an influence of the topology, as meta-sub-
stituted 7Ce and 7Cf showed the highest relative affinity
values.
In the case of the trivalent systems (Table 3, Entries 9–
14), we bring attention to an influence of the electronic den-
sity of the aromatic core on the binding recognition proper-
ties. Thus, the thioglycoside-containing 7Ff was higher in
affinity than its O-glycoside-containing homologue 7Fe, but
this trend was inverted in the electronic-rich neoglycoconju-
gates 7Ee, 7Ef, 7Ge, and 7Gf, where the higher values were
observed for the O-glycosides 7Ee and 7Gf.
In the case of the O-glycoside-containing tetravalent sys-
tems (7He, 8Jh, and 8Kh, Table 3, Entries 15, 20, and 22)
we observed that the aromatic-centered 7He had a higher
binding affinity than that of the non-aromatic 8Jh and 8Kh.
This fact is in line with the previously observed tendency
for divalent systems, where the aromatic neoglycoconjugates
showed better binding properties. These data, together with
the higher relative affinity values of the cyclic sugar deriva-
tives 8Lg and 8Lh relative to that of the open-chain manni-
tol derivatives 8Mg and 8Mh, indicates that rigid systems
are better binders than more conformationally flexible com-
pounds. However, we could not be discard the interactions
of the aromatic rings with Con A residues to explain this
increased binding.[20] Previous contributions by others have
diverse, α-Man-containing, homogeneous neoglycoconju-
gates possessing aliphatic-, aromatic-, and carbohydrate-
centered architectures using CuAAC reactions as a covalent
ligating tool for the assembly of clickable anomeric α-Man
derivatives onto suitable, complementary, functionalized
scaffolds. The methodology is highly efficient and versatile
showing its value as a general and convenient way to con-
nect multiple units of a sugar residue to a central core for
the rapid generation of tailor-made neoglycoconjugates dif-
fering in structural factors such as the sugar densities (val-
ency), topology (substitution pattern), nature of the linker,
and electronic characteristics of the core. The ELISA-type
assays performed with the novel, synthesized neoglycocon-
jugates to evaluate the influence of those structural param-
eters allowed us to obtain relevant information. Thus, the
valency, electronic characteristics of the core, and linker
were the most important parameters that determine the in-
hibitory properties of the neoglycoconjugates.
Experimental Section
General: TLC was performed on Merck Silica Gel 60 F254 alumi-
num sheets. Reagents used for developing plates include ceric sul-
fate (1% w/v) and ammonium sulfate (2.5% w/v) in 10% (v/v)
aqueous sulfuric acid, iodine, ethanolic sulfuric acid (10% v/v), and
UV light when applicable. Flash column chromatography was per-
clearly established the positive effect of the presence of an formed on Silica Gel Scharlau (230–400 mesh, ASTM). Melting
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F. Santoyo-Gonzalez et al.
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points were measured with a Gallenkamp melting point apparatus
and are uncorrected. Optical rotations were recorded with a Per-
kin–Elmer 141 polarimeter at room temp. IR spectra were recorded
with a Satellite Mattson FTIR. 1H and 13C NMR spectra were
recorded at room temp. with a Bruker (300 or 400 MHz) spectrom-
eter. Chemical shifts are given in ppm and referenced to internal
CDCl3. J values are given in Hz. FAB mass spectra were recorded
with a Fissons VG Autospec-Q spectrometer, using m-nitrobenzyl
alcohol or thioglycerol as the matrix. MALDI-TOF mass spectra
were recorded with a Bruker Autoflex spectrometer using DHB
or HCCA as the matrix. Propargyl bromide, α,α-dichloro-m-xilene,
cyanuric chloride, and 1,2,4,5-tetrakis(bromomethyl)benzene were
purchased from Aldrich. Compounds 1a,[22,23] 1b,[24] 2c,[25] 2d,[26]
3B–3D,[27] 3F,[28] 3G,[29] 3H,[30] 3I,[31] 4J,[32] and 4L[17] were obtained
following the procedures described in the literature.
hexane (1:3) giving 4M as a syrup (1.71 g, 74%): IR (KBr): ν =
˜
3286, 2889, 2112, 1483, 1327, 1095 cm–1. 1H NMR (CDCl3,
300 MHz): δ = 4.39 (d, J = 2.4 Hz, 4 H), 4.35 (d, J = 2.4 Hz, 2 H),
4.33 (d, J = 2.4 Hz, 2 H), 4.23 (d, J = 2.3 Hz, 4 H), 3.85 (several
m, 8 H), 2.45 (m, 6 H) ppm. 13C NMR (CDCl3, 75 MHz): δ =
80.23, 79.9, 79.6, 78.1, 77.3, 74.5, 68.1, 59.9, 58.5, 56.9 ppm.
HRMS (FAB+): calcd. for C24H26O6Na [M + Na]+ 433.1627;
found 433.1621.
General Procedure for the Cycloaddition Reactions. Method A: A
solution in toluene (10 mL) of the polyazide scaffold 3A–3I
(1 equiv.), the corresponding complementary propargyl sugar 1a–b
(1.2 equiv. per reactive group of the scaffold), and the Cu catalyst
[(EtO)3P·CuI][16] (0.1 equiv. per reactive group of the scaffold) was
irradiated at 800 W and 90 °C in a Milestone Star Microwave
Labstation until TLC or the IR spectrum of the reaction mixture
showed complete disappearance of the starting material (see
Table 1 for reaction times). Evaporation of the solvent yielded a
crude material that was purified by column chromatography.
2,4,6-Tris(2-azidoethoxy)-1,3,5-triazine (3E): To a cold solution of
cyanuric chloride (1.87 g, 10 mmol) and 2-azido ethanol (5.2 g,
60 mmol) in dry acetonitrile (25 mL) was added dropwise a solu-
tion of DIPEA (7 mL) in acetonitrile (10 mL). The reaction mix-
ture was maintained at room temp. for 2 d. After the solvent was
evaporated, the crude product was dissolved in chloroform
(150 mL) and washed with H2O (50 mL). The organic phase was
dried (Na2SO4) and filtered, and the solvents were evaporated. Col-
Method B: A solution in toluene (10 mL) of the polyazide or poly-
alkyne 3A–3I (1 equiv.), the propargyl sugar derivative 1a–b
(1.2 equiv. per reactive group of the scaffold), the Cu catalyst [(EtO)
3P·CuI][16] (0.1 equiv. per reactive group of the scaffold), and DI-
PEA (3.0 equiv. per reactive group of the scaffold) was irradiated
at 800 W and 90 °C in a Milestone Star Microwave Labstation until
TLC or the IR spectrum of the reaction mixture showed complete
disappearance of the starting material (see Table 1 for reaction
times). Evaporation of the solvent yielded a crude material that
was purified by column chromatography.
umn chromatography (CH2Cl2/diethyl ether, 20:1) gave 3E (2.34 g,
1
70%) as a liquid. IR (film): ν = 2105, 1554, 1329, 1294 cm–1. H
˜
NMR (CDCl3, 300 MHz): δ = 4.59 (t, J = 5.1 Hz, 1 H, CH2O), 3.68
(t, J = 5.1 Hz, 6 H, CH2N3) ppm. 13C NMR (CDCl3, 75 MHz): δ
= 171.8, 67.3, 49.4 ppm.
Methyl 2,3,4,6-Tetra-O-propargyl-α-D-glucopyranoside (4K): NaH
Method C: A solution in DMF (10 mL) of the polyazide scaffold
3I (1 equiv.), the corresponding complementary propargyl sugar
derivative 1b (1.2 equiv. per reactive group of the scaffold), the Cu
catalyst [(EtO)3P·CuI][16] (0.1 equiv. per reactive group of the scaf-
fold), and DIPEA (3.0 equiv. per reactive group of the scaffold)
was heated at 80 °C until TLC and the IR spectrum of the reaction
mixture showed complete disappearance of the starting material
(see Table 1 for reaction times). Evaporation of the solvent yielded
a crude material that was purified by column chromatography.
(371 mg, 15.46 mmol) was added to a solution of methyl α--gluco-
pyranoside (1 g, 5.49 mmol) in anhydrous DMF (25 mL). The sus-
pension was magnetically stirred for 30 min at room temp. Propar-
gyl bromide (7.7 mL, 103 mmol) was then added dropwise, and the
reaction mixture was maintained for 16 h. After the mixture was
cooled, MeOH (10 mL) was added, and the solvents were removed
after 30 min. The addition of H2O (20 mL) to the resulting crude
material was followed by extraction with toluene/diethyl ether (3:1,
2ϫ50 mL). The organic phase was dried (Na2SO4) and filtered,
and the solvents were evaporated under vacuum The crude material
was purified by column chromatography EtOAc/hexane (1:3) giving
4K as a syrup (752 mg, 85%): [α]2D0 = +80 (c = 1 in chloroform);
Method D: A solution in toluene (10 mL) of the polyalkyne scaffold
4J–4M (1 equiv.), the corresponding complementary azide sugar
derivative 2c–d (1.2 equiv. per reactive group of the scaffold), and
the Cu catalyst [(EtO)3P·CuI][16] (0.1 equiv. per reactive group of
the scaffold) was refluxed until TLC or the IR spectrum of the
reaction mixture showed complete disappearance of the starting
material (see Table 2 for reaction times). Evaporation of the solvent
yielded a crude material that was purified by column chromatog-
raphy.
IR (KBr): ν = 3290, 2923, 2867, 2116, 1728, 1446, 1355, 1267, 1158,
˜
1
1043 cm–1. H NMR (CDCl3, 300 MHz): δ = 4.90 (d, J = 3.5 Hz,
1 H, H-1), 4.36 (several m, 8 H, CH2CϵCH), 3.84 (t, J = 9.2 Hz,
1 H), 3.82 (m, 2 H, H-6,6), 3.71 (m, 1 H, H-5), 3.62 (dd, J = 9.6,
3.6 Hz, 1 H, H-2), 3.51 (m, 1 H, H-4), 3.41 (s, 1 H, OMe), 2.47 (m,
4 H, CϵCH) ppm. 13C NMR (CDCl3, 75 MHz): δ = 97.9, 81.2,
80.0, 79.7, 79.3, 79.1, 76.5, 75.0, 74.3, 74.2, 69.4, 68.0, 60.3, 60.1,
58.6, 58.6, 55.2 ppm. HRMS (FAB+): calcd. for C19H22O6Na [M
+ Na]+ 369.1314; found 369.1315.
1,2-Bis{4-(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyloxymethyl)-
1H-1,2,3-triazol-1-yl}ethane (5Aa): Obtained from 1a (463 mg) and
3A (56 mg) following the general procedure (Method A). Column
chromatography gave 5Aa as a solid (411 mg, 93%); m.p. 59–61 °C.
1,2,3,4,5,6-Hexakis-O-propargyl-D-mannitol (4M): NaH (1.25 g,
49.40 mmol) was added to a solution of -mannitol (500 mg, [α]2D0 = +37 (c = 2 in chloroform); IR (KBr): ν = 3144, 1746, 1372,
˜
1
2.56 mmol) in anhydrous DMF (25 mL). The suspension was mag-
netically stirred for 30 min at room temp. Propargyl bromide (80%
toluene solution, 24.5 mL, 165 mmol) was then added dropwise,
and the reaction mixture was maintained for 16 h. After the mix-
ture was cooled, MeOH (10 mL) was added, and the solvents were
removed after 30 min. The addition of H2O (20 mL) to the re-
sulting crude material was followed by extraction with toluene/di-
ethyl ether (3:1, 2ϫ50 mL). The organic phase was dried (Na2SO4)
and filtered, and the solvents were evaporated under vacuum The
crude material was purified by column chromatography EtOAc/
1230, 1135, 1048 cm–1. H NMR (CDCl3, 300 MHz): δ = 7.43 (s,
2 H, H-5 triazole), 5.30–5.25 (m, 4 H, H-3,4), 5.19 (s, 2 H, H-2),
4.96 (s, 4 H, CH2N), 4.94 (s, 2 H, H-1), 4.80 (d, J = 12.3 Hz, 2 H,
CH2O), 4.65 (d, J = 12.3 Hz, 2 H, CH2O), 4.29 (dd, J = 12.2,
5.0 Hz, 2 H, H-6), 4.11 (dd, J = 12.3, 2.4 Hz, 2 H, H-6), 4.06 (m,
2 H, H-5), 2.15, 2.12, 2.04, 1.98 (4 s, 24 H, 8 Ac) ppm. 13C NMR
(CDCl3, 75 MHz): δ = 170.7, 170.1, 170.0, 169.8, 144.0, 124.3, 96.9,
69.5, 69.1, 68.8, 66.2, 62.5, 60.7, 49.6, 20.9, 20.8, 20.7, 20.7 ppm.
HRMS (FAB+): calcd. for C36H48N6O20 [M + Na]+ 907.2821;
found 907.2824.
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Click Multivalent Homogeneous Neoglycoconjugates
1,2-Bis{4-(2,3,4,6-tetra-O-acetyl-α- -mannopyranosylthiomethyl)-
D
(482 mg) and 3C (99 mg) following the general procedure (Method
B). Column chromatography (EtOAc) gave 5Cb as a solid (427 mg,
1H-1,2,3-triazol-1-yl}ethane (5Ab): Obtained from 1b (482 mg) and
3A (56 mg) following the general procedure, (Method B). Column
chromatography (EtOAc/MeOH, 20:1) gave 5Ab as a solid (352 mg,
86%); m.p. 87–88 °C. [α]2D0 = +112 (c = 1 in chloroform); IR (KBr):
1
ν = 1749, 1371, 1228, 1050 cm–1. H NMR (CDCl , 300 MHz): δ
˜
3
77%); m.p. 88–91 °C. [α]2D0 = +158 (c = 1 in chloroform); IR (KBr):
= 7.48 (s, 2 H, H-5 triazole), 7.25, 7.22 (br. s, 4 H, C6H4), 5.51 (s,
4 H, CH2N), 5.36–5.21 (m, 8 H, H-1,2,3,4), 4.36 (ddd, J = 9.8, 5.0,
2.4 Hz, 2 H, H-5), 4.28 (dd, J = 12.1, 5.1 Hz, 2 H, H-6), 4.04 (dd,
J = 12.1, 2.1 Hz, 2 H, H-6), 3.95 (d, J = 14.6 Hz, 2 H, CH2S), 3.84
1
ν = 1748, 1371, 1228, 1050 cm–1. H NMR (CDCl , 300 MHz): δ
˜
3
= 7.34 (s, 2 H), 5.33 (t, J = 9.8 Hz, 2 H), 5.28 (d, J = 3.3 Hz, 2 H),
5.26 (s, 2 H), 5.21 (dd, J = 10.0, 3.2 Hz, 2 H), 4.91 (AB system, 4
H), 4.37 (m, 2 H), 4.29 (dd, J = 12.2, 5.0 Hz, 2 H), 4.09 (m, 2 H), (d, J = 14.6 Hz, 2 H, CH2S), 2.14, 2.09, 2.05, 1.98 (4 s, 24 H, 8 Ac)
3.91 (d, J = 14.6 Hz, 2 H) 3.80 (d, J = 14.5 Hz, 2 H), 2.15, 2.11,
ppm. 13C NMR (CDCl3, 75 MHz): δ = 170.6, 169.9, 169.8, 169.6,
2.06, 1.99 (4 s, 24 H) ppm. 13C NMR (CDCl3, 75 MHz): δ = 170.5, 144.6, 135.7, 132.1, 130.0, 128.5, 127.5, 122.1, 81.7, 70.7, 69.5, 69.3,
169.9, 169.7, 169.6, 144.3, 123.3, 81.4, 70.6, 69.5, 69.2, 66.1, 62.3,
66.2, 62.2, 53.7, 24.8, 20.8, 20.7, 20.6 ppm. HRMS (FAB+): calcd.
49.5, 24.4, 20.8, 20.7, 20.6, 20.5 ppm. HRMS (MALDI-TOF): for C42H52N6O18S2 [M + Na]+ 1015.2677; found 1015.2670.
calcd. for C36H49N6O20S2 [M + H]+ 917.2540; found 917.2050.
1,4-Bis{4-(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyloxymethyl)-
1,2-Bis{4-(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyloxymethyl)-
1H-1,2,3-triazole-1-ylmethyl}benzene (5Da): Obtained from 1a
(463 mg) and 3D (94 mg) following the general procedure (Method
A). Column chromatography (EtOAc) gave 5Da as a solid (355 mg,
74%); m.p. 140–142 °C. [α]2D0 = +45 (c = 1 in chloroform); IR
1H-1,2,3-triazol-1-ylmethyl}benzene (5Ba): Obtained from 1a
(4623 mg) and 3B (482 mg) following the general procedure
(Method A). Column chromatography (EtOAc) gave 5Ba as a
syrup (432 mg, 90%): [α]2D0 = +39 (c = 1 in MeOH); IR (KBr): ν =
˜
(KBr): ν = 3142, 1755, 1373, 1245, 1135, 1079 cm–1. 1H NMR
˜
1747, 1438, 1372, 1227, 1135, 1048 cm–1
.
1H NMR (CDCl3,
(CDCl3, 300 MHz): δ = 7.58 (s, 2 H, H-5 triazole), 7.32 (s, 4 H,
C6H4), 5.56 (s, 4 H, CH2N), 5.31–5.27 (m, 4 H, H-3,4), 5.22 (br. s,
2 H, H-2), 4.95 (s, 2 H, H-1), 4.82 (d, J = 12.3 Hz, 2 H, CH2O),
4.67 (d, J = 12.3 Hz, 2 H, CH2O), 4.27 (dd, J = 12.5, 5.3 Hz, 2 H,
H-6), 4.15–4.00 (m, 4 H, H-5,6), 2.15, 2.10, 2.03, 1.98 (4 s, 24 H,
8 Ac) ppm. 13C NMR (CDCl3, 75 MHz): δ = 170.5, 169.8, 169.7,
169.5, 143.9, 135.1, 128.7, 122.9, 96.8, 69.3, 68.9, 68.6, 66.1, 62.2,
60.9, 53.5, 20.7, 20.6, 20.5 ppm. HRMS (FAB+): calcd. for
C42H52N6O20 [M + Na]+ 983.3134; found 983.3140.
300 MHz): δ = 7.59 (s, 2 H, H-5 triazole), 7.42 (dd, J = 5.7, 3.4 Hz,
1 H, C6H4), 7.29 (dd, J = 5.6, 3.4 Hz, 2 H, C6H4), 5.68 (s, 4 H,
CH2N), 5.30–5.25 (m, 4 H, H-3,4), 5.22 (dd, J = 2.8, 1.6 Hz, 2 H,
H-2), 4.95 (d, J = 1.6 Hz, 2 H, H-1), 4.82 (d, J = 12.4 Hz, 2 H,
CH2O), 4.67 (d, J = 12.4 Hz, 2 H, CH2O), 4.28 (dd, J = 12.4,
5.2 Hz, 2 H, H-6), 4.15–4.00 (m, 4 H, H-5,6), 2.14, 2.11, 2.03, 1.98
(4 s, 24 H, 8 Ac) ppm. 13C NMR (CDCl3, 75 MHz): δ = 170.6,
169.9, 169.8, 169.6, 143.9, 133.2, 130.4, 129.9, 123.2, 96.9, 69.4,
69.0, 68.7, 66.0, 62.3, 60.9, 51.2, 21.0, 20.8, 20.7, 20.6 ppm. HRMS
(FAB+): calcd. for C42H52N6O20 [M + Na]+ 983.3134; found 1,4-Bis{4-(2,3,4,6-tetra-O-acetyl-α-
D-mannopyranosylthiomethyl)-
983.3138.
1H-1,2,3-triazol-1-ylmethyl}benzene (5Db): Obtained from 1b
(482 mg) and 3D (94 mg)following the general procedure (Method
B). Column chromatography (EtOAc) gave 5Db as a solid (337 mg,
1,3-Bis{4-(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyloxymethyl)-
1H-1,2,3-triazole-1-ylmethyl}benzene (5Bb): Obtained from 1b
(482 mg) and 3B (94 mg) following the general procedure (Method
C). Column chromatography (EtOAc) gave 5Bb as a solid (377 mg,
68%); m.p. 155–157 °C. [α]2D0 = +117 (c = 1 in chloroform); IR
1
(film): ν = 1747, 1370, 1228, 1108, 1050 cm–1. H NMR (CDCl ,
˜
3
75%); m.p. 88–89 °C. [α]2D0 = +185 (c = 1 in chloroform); IR (KBr):
300 MHz): δ = 7.50 (s, 2 H, H-5 triazole), 7.28 (s, 4 H, C6H4), 5.52
(s, 4 H, CH2N), 5.32 (t, J = 9.9 Hz, 2 H, H-4), 5.29 (s, 4 H, H-1,2),
5.22 (dd, J = 10.0, 2.9 Hz, 2 H, H-3), 4.36 (ddd, J = 9.8, 5.0, 2.0 Hz,
2 H, H-5), 4.28 (dd, J = 12.2, 5.1 Hz, 2 H, H-6), 4.03 (dd, J = 12.1,
2.1 Hz, 2 H, H-6), 3.93 (d, J = 14.6 Hz, 2 H, CH2S), 3.82 (d, J =
14.6 Hz, 2 H, CH2S), 2.13, 2.08, 2.05, 1.98 (4 s, 24 H, 8 Ac) ppm.
13C NMR (CDCl3, 75 MHz): δ = 170.4, 169.7, 169.6, 144.4, 135.2,
132.0, 131.9, 128.7, 128.6, 128.5, 128.3, 122.1, 81.6, 70.5, 69.4, 69.1,
66.1, 62.1, 53.5, 24.7, 20.9, 20.7, 20.6, 20.4 ppm. HRMS (FAB+):
calcd. for C42H52N6O18S2 [M + Na]+ 1015.2677; found 1015.2679.
ν = 1750, 1371, 1229, 1050 cm–1. H NMR (CDCl , 300 MHz): δ
1
˜
3
= 7.47 (s, 2 H), 7.40–7.23 (m, 4 H), 5.62 (s, 4 H), 5.36–5.20 (m, 8
H), 4.36 (ddd, J = 9.5, 5.0, 2.3 Hz, 2 H), 4.28 (dd, J = 12.2, 5.0 Hz,
2 H), 4.06 (dd, J = 12.2, 2.2 Hz, 2 H), 3.94 (d, J = 14.6 Hz, 2 H),
3.83 (d, J = 14.6 Hz, 2 H), 2.13, 2.09, 2.05, 1.98 (4 s, 24 H) ppm.
13C NMR (CDCl3, 75 MHz): δ = 170.5, 169.8, 179.7, 169.6, 144.5,
133.2, 130.4, 129.7, 122.3, 81.6, 70.6, 69.5, 69.2, 66.2, 62.2, 51.2,
24.7, 20.8, 20.7, 20.6, 20.5 ppm. HRMS (MALDI-TOF): calcd. for
C42H52N6O18S2 [M + Na]+ 1015.268; found 1015.295.
1,3-Bis{4-(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyloxymethyl)-
1,3,5-Tris{2-[4-(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyloxymeth-
1H-1,2,3-triazole-1-ylmethyl}benzene (5Ca): Obtained from 1a
(463 mg) and 3C (99 mg) following the general procedure (Method
A). Column chromatography (EtOAc) gave 5Ca as a solid (384 mg,
yl)-1H-1,2,3-triazol-1-yl]-ethoxy}benzene (5Ea): Obtained from 1a
(695 mg) and 3E (166 mg) following the general procedure (Method
A). Column chromatography (EtOAc Ǟ EtOAc/MeOH, 50:1) gave
5Ea as a syrup (530 mg, 71%): [α]2D0 = +55 (c = 1 in chloroform);
80%); m.p. 81–83 °C. [α]2D0 = +16 (c = 1 in MeOH); IR (KBr): ν =
˜
1746, 1457, 1371, 1227, 1135, 1048 cm–1
.
1H NMR (CDCl3,
IR (KBr): ν = 1746, 1370, 1226, 1047 cm–1. 1H NMR (CDCl3,
˜
300 MHz): δ = 7.55 (s, 2 H, H-5 triazole), 7.41 (t, J = 7.5 Hz, 1 H,
C6H4), 7.30–7.22 (m, 3 H, C6H4), 5.60 (s, 4 H, CH2N), 5.31–5.26
(m, 4 H, H-3,4), 5.21 (br. d, J = 1.7 Hz, 2 H, H-2), 4.94 (d, J =
1.7 Hz, 2 H, H-1), 4.84 (d, J = 12.4 Hz, 2 H, CH2O), 4.67 (d, J =
12.4 Hz, 2 H, CH2O), 4.28 (dd, J = 12.2, 5.1 Hz, 2 H, H-6), 4.12–
4.00 (m, 4 H, H-5,6), 2.14, 2.10, 2.03, 1.99 (4 s, 24 H, 8 Ac) ppm.
13C NMR (CDCl3, 75 MHz): δ = 170.8, 170.1, 169.8, 144.0, 135.7,
130.2, 128.5, 127.6, 123.1, 97.0, 69.6, 69.1, 68.8, 66.2, 62.4, 61.2,
53.9, 20.9, 20.8, 20.7 ppm. HRMS (FAB+): calcd. for C42H52N6O20
[M + Na]+ 983.3134; found 983.3136.
300 MHz): δ = 7.76 (br. s, 3 H, H-5 triazole), 6.09 (s, 3 H, ArH),
5.35–5.25 (m, 6 H, H-3,4), 5.24 (s, 3 H, H-2), 4.96 (s, 3 H, H-1),
4.85 (d, J = 12.5 Hz, 3 H, OCH2), 4.68 (d, J = 12.3 Hz, 3 H,
OCH2), 4.75 (t, J = 4.7 Hz, 6 H, OCH2CH2N), 4.34 (t, J = 4.5 Hz,
6 H, OCH2CH2N), 4.29 (dd, J = 12.3, 5.1 Hz, 3 H, H-6), 4.10 (d,
J = 12.3 Hz, 3 H, H-6), 4.08 (m, 3 H, H-5), 2.15, 2.12, 2.03, 1.98
(4 s, 36 H, MeCO) ppm. 13C NMR (CDCl3, 75 MHz): δ = 170.8,
170.2, 170.0, 169.8 (CO), 159.9 (Ar), 143.6 (C-4 triazole), 124.2 (C-
5 triazole), 97.0, (C-1), 94.9 (Ar), 69.4 (C-2), 69.1 (C-3), 68.8 (C-
5), 66.4 (C-4), 66.0 (OCH2CH2N), 62.4 (C-6), 61.1 (CH2O), 49.8
(CH2N), 20.9, 20.8, 20.8 (MeCO) ppm. HRMS (FAB+): calcd. for
C63H81N9O33 [M + Na]+ 1514.4834; found 1514.5100.
1,3-Bis{4-(2,3,4,6-tetra-O-acetyl-α-
D-mannopyranosylthiomethyl)-
1H-1,2,3-triazole-1-ylmethyl}benzene (5Cb): Obtained from 1b
Eur. J. Org. Chem. 2009, 2441–2453
© 2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.eurjoc.org
2447

F. Santoyo-Gonzalez et al.
FULL PAPER
1,3,5-Tris{2-[4-(2,3,4,6-tetra-O-acetyl-α-
D
-mannopyranosylthiometh- 1,3,5-Tris{4-(2,3,4,6-tetra-O-acetyl-α-
D
-mannopyranosylthiometh-
yl)-1H-1,2,3-triazol-1-yl]-ethoxy}benzene (5Eb): Obtained from 1b
(725 mg) and 3E (166 mg) following the general procedure (Method
yl)-1H-1,2,3-triazol-1-ylmethyl}-2,4,6-trimethoxybenzene (5Gb):
Obtained from 1b (725 mg) and 3G (166 mg) following the general
A). Column chromatography (EtOAc Ǟ EtOAc/MeOH, 50:1) gave procedure (Method A). Column chromatography (EtOAc Ǟ
5Eb as a syrup (705 mg, 92%): [α]2D0 = +133 (c = 1 in chloroform);
EtOAc/MeOH, 50:1) gave 5Gb as a foamy solid (765 mg, 100%):
IR (film): ν = 1746, 1226, 1049 cm–1. 1H NMR (CDCl , 300 MHz): [α]2D0 = +127 (c = 1 in chloroform); IR (film): ν = 1747, 1226, 1103,
˜
˜
3
δ = 7.67 (s, 3 H, H-5 triazole), 6.03 (s, 3 H, ArH), 5.36–5.30 (m, 9
1048 cm–1. 1H NMR (CDCl3, 300 MHz): δ = 7.61 (s, 3 H, H-5
H, H-1,2,4), 5.23 (dd, J = 10.0, 2.8 Hz, 3 H, H-3), 4.73 (t, J = triazole), 5.55 (s, 6 H, CH2N), 5.37–5.20 (m, 12 H, H-1,2,3,4), 4.38
4.5 Hz, 6 H, OCH2), 4.40–4.25 (m, 12 H, H-5,6, CH2N), 4.07 (br. (m, 3 H, H-5), 4.33 (dd, J = 12.0, 4.9 Hz, 3 H, H-6), 4.09 (dd, J =
d, J = 10.0 Hz, 3 H, H-6), 3.96 (d, J = 14.5 Hz, 3 H, CH2S), 3.68
12.0, 1.8 Hz, 3 H, H-6), 3.96 (d, J = 14.5 Hz, 3 H, CH2S), 3.84 (d,
(d, J = 14.5 Hz, 3 H, CH2S), 2.12, 2.09, 2.05, 1.98 (4 s, 36 H, J = 14.5 Hz, 3 H, CH2S), 3.76 (s, 9 H, MeO), 2.15, 2.10, 2.06, 1.98
MeCO) ppm. 13C NMR (CDCl3, 75 MHz): δ = 170.7, 170.0, 169.9, (4 s, 36 H, MeCO) ppm. 13C NMR (CDCl3, 75 MHz): δ = 170.7,
169.8 (CO), 159.9 (Ar), 144.2 (C-4 triazole), 123.3 (C-5 triazole),
170.0, 169.9, 169.7 (CO), 160.8 (Ar), 144.5 (C-4 triazole), 122.6
95.0 (Ar), 81.7 (C-1), 70.6, 69.7, 69.3, 66.3 (C-2,3,4,5), 66.5 (Ar), 119.9 (C-5 triazole), 81.8 (C-1), 70.8 (C-3), 69.6 (C-2), 69.4
(CH2O), 62.4 (C-6), 49.8 (CH2N), 24.8 (CH2S), 20.9, 20.8, 20.7, (C-5), 66.3 (C-4), 63.2 (OMe), 62.4 (C-6), 43.9 (CH2N), 24.9
20.6 ppm. HRMS (FAB+): calcd. for C63H81N9O30S3 [M + Na]+
1562.415; found 1562.4020.
(CH2S), 21.1, 20.9, 20.8 (MeCO) ppm. HRMS (FAB+): calcd. for
C63H81N9O30S3 [M + Na]+ 1562.415; found 1562.38.
1,3,5-Tris{4-(2,3,4,6-tetra-O-acetyl-α-
yl)-1H-1,2,3-triazol-1-ylmethyl}-2,4,6-trimethylbenzene (5Fa): Ob-
tained from 1a (695 mg) and 3F (143 mg) following the general 1a (925 mg) and 3H (149 mg) following the general procedure
D
-mannopyranosyloxymeth- 1,2,4,5-Tetrakis{4-(2,3,4,6-tetra-O-acetyl-α-
D-mannopyranosyloxy-
methyl)-1H-1,2,3-triazol-1-ylmethyl}benzene (5Ha): Obtained from
procedure (Method B). Column chromatography (EtOAc Ǟ
EtOAc/MeOH, 15:1) gave 5Fa as a foamy solid (560 mg, 78%):
(Method B). Column chromatography (EtOAc Ǟ EtOAc/MeOH,
10:1) gave 5Ha as a solid (780 mg, 85%); m.p. 109–111 °C. [α]2D0
+54 (c = 1 in chloroform); IR (KBr): ν = 3143, 1750, 1438, 1372,
=
[α]2D0 = +41.3 (c = 1, chloroform); IR (KBr): ν = 1749, 1372, 1234,
˜
˜
1049 cm–1. 1H NMR (CDCl3, 300 MHz): δ = 7.41 (s, 3 H), 5.72 (s, 1239, 1134, 1049 cm–1. H NMR (CDCl3, 300 MHz): δ = 7.70 (s,
1
6 H), 5.40–5.25 (m, 6 H), 5.19 (s, 3 H), 4.94 (s, 1 H), 4.81 (d, J = 4 H, H-5 triazole), 7.15 (s, 2 H, Ar), 5.68 (s, 8 H, CH2N), 5.35–
12.4 Hz, 3 H), 4.64 (d, J = 12.3 Hz, 3 H), 4.29 (dd, J = 12.2, 5.1 Hz,
3 H), 4.20–4.05 (m, 6 H), 2.47 (s, 9 H), 2.15, 2.11, 2.04, 1.99 (4 s,
36 H) ppm. 13C NMR (CDCl3, 75 MHz): δ = 170.5, 169.9, 169.8,
169.5, 143.5, 139.9, 130.5, 122.3, 96.7, 69.3, 68.9, 68.6, 66.0, 62.2,
60.8, 49.0, 20.9, 20.7, 20.6, 20.5, 16.7 ppm. HRMS (FAB+): calcd.
for C63H81N9O30 [M + Na]+ 1466.4987; found 1466.4984.
5.25 (m, 8 H, H-3,4), 5.20 (s, 4 H, H-2), 4.98 (s, 4 H, H-1), 4.83
(d, J = 12.3 Hz, 4 H, CH2O), 4.68 (d, J = 12.3 Hz, 4 H, CH2O),
4.29 (dd, J = 12.2, 4.9 Hz, 4 H, H-6), 4.15–4.00 (m, 8 H, H-5,6),
2.14, 2.11, 2.04, 1.98 (4 s, 48 H, 16 Ac) ppm. 13C NMR (CDCl3,
75 MHz): δ = 170.6, 169.9, 169.8, 169.6, 144.0, 134.9, 132.0, 123.6,
96.7, 69.3, 68.9, 68.6, 65.9, 62.2, 60.6, 50.5, 20.7, 20.6, 20.5 ppm.
HRMS (FAB+): calcd. for C78H98N12O40 [M + Na]+ 1865.5901;
found 1865.5856.
1,3,5-Tris{4-(2,3,4,6-tetra-O-acetyl-α-
D-mannopyranosylthiometh-
yl)-1H-1,2,3-triazol-1-ylmethyl}-2,4,6-trimethylbenzene (5Fb): Ob-
tained from 1b (725 mg) and 3F (143 mg) following the general 1,2,4,5-Tetrakis{4-(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosylthio-
procedure (Method B). Column chromatography (EtOAc) gave 5Fb
methyl)-1H-1,2,3-triazol-1-methyl}benzene (5Hb): Obtained from
1b (965 mg) and 3H (149 mg) following the general procedure
as a solid (580 mg, 78%); m.p. 102–105 °C. [α]2D0 = +143 (c = 1 in
chloroform); IR (KBr): ν = 1749, 1371, 1229, 1049 cm–1. 1H NMR (Method B). Column chromatography (EtOAc/MeOH, 50:1) gave
˜
(CDCl3, 300 MHz): δ = 7.31 (s, 3 H), 5.66 (s, 6 H), 5.33 (t, J =
9.8 Hz, 3 H), 5.28 (s, 1 H), 5.26 (d, J = 3.4 Hz, 3 H), 5.21 (dd, J =
9.9, 3.3 Hz, 3 H), 4.37 (m, 3 H), 4.30 (dd, J = 12.0, 5.1 Hz, 3 H),
5Hb as a solid (735 mg, 77%); m.p. 112–115 °C. [α]2D0 = +144 (c =
1
1 in chloroform); IR (KBr): ν = 1748, 1372, 1230, 1050 cm–1. H
˜
NMR (CDCl3, 300 MHz): δ = 7.51 (s, 4 H), 4.14 (s, 2 H), 5.60 (s,
4.08 (d, J = 12.0 Hz, 3 H), 3.93 (d, J = 14.6 Hz, 3 H), 3.80 (d, J = 8 H), 5.34 (t, J = 9.8 Hz, 4 H), 5.31 (s, 4 H), 5.27 (d, J = 3.3 Hz,
14.6 Hz, 3 H), 2.41 (s, 9 H), 2.14, 2.09, 2.05, 1.99 (4 s, 36 H) ppm. 4 H), 5.21 (dd, J = 10.0, 3.3 Hz, 4 H), 4.35 (m, 4 H), 4.28 (dd, J =
13C NMR (CDCl3, 75 MHz): δ = 170.5, 169.8, 169.7, 169.5, 144.1, 12.1, 4.8 Hz, 4 H), 4.08 (dd, J = 12.1, 2.1 Hz, 4 H), 3.95 (d, J =
139.7, 130.5, 121.4, 81.5, 70.6, 69.4, 69.2, 66.0, 62.1, 49.0, 24.6,
20.8, 20.6, 20.5, 20.5, 16.5 ppm. HRMS (MALDI-TOF): calcd. for
C63H81N9O27S3 [M + H]+ 1492.448; found 1492.434.
14.7 Hz, 4 H), 3.83 (d, J = 14.6 Hz, 4 H), 2.13, 2.10, 2.06, 1.98 (4
s, 48 H) ppm. 13C NMR (CDCl3, 75 MHz): δ = 170.6, 169.9, 169.7,
169.6, 144.7, 134.8, 132.1, 122.6, 81.5, 70.6, 69.4, 69.2, 66.1, 62.1,
50.4, 24.5, 20.8, 20.7, 20.6, 20.5 ppm. HRMS (FAB+): calcd. for
C78H98N12O36S4 [M + Na]+ 1929.499; found 1929.496.
1,3,5-Tris{4-(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyloxymeth-
yl)-1H-1,2,3-triazol-1-ylmethyl}-2,4,6-trimethoxybenzene (5Ga):
Obtained from 1a (695 mg) and 3G (166 mg) following the general
procedure (Method A). Column chromatography (EtOAc Ǟ
1,2,3,4,5,6-Hexakis{4-(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyl-
oxymethyl)-1H-1,2,3-triazol-1-ylmethyl}benzene (5Ia): Obtained
from 1a (695 mg) and 3I (102 mg) following the general procedure
(Method B). Column chromatography (Cl2CH2/MeOH, 100:1 Ǟ
EtOAc/MeOH, 50:1) gave 5Ga as a syrup (580 mg, 79%): [α]2D0
=
+39 (c = 1 in chloroform); IR (film): ν = 2948, 1750, 1586, 1456,
˜
1368, 1220, 1330, 1043 cm–1. 1H NMR (CDCl3, 300 MHz): δ =
25:1) gave 5Ia as a solid (547 mg, 77%); m.p. 162–164 °C. [α]2D0
=
7.75 (s, 3 H, H-5 triazole), 5.60 (s, 6 H, CH2N), 5.31 (m, 6 H, H- +56 (c = 1 in chloroform); IR (KBr): ν = 1749, 1371, 1228, 1135,
˜
3,4), 5.21 (s, 3 H, H-2), 4.96 (s, 3 H, H-1), 4.82 (d, J = 12.3 Hz, 3
H, CH2O), 4.65 (d, J = 12.3 Hz, 3 H, CH2O), 4.30 (dd, J = 12.4,
1080, 1048 cm–1. 1H NMR (CDCl3, 300 MHz): δ = 7.56 (s, 1 H,
H-5), 5.85 (s, 2 H, CH2N), 5.27 (m, 2 H, H-3,4), 5.16 (s, 1 H, H-
5.0 Hz, 3 H, H-6), 4.10 (m, 6 H, H-5,6), 3.78 (s, 9 H, MeO), 2.15, 2), 4.90 (s, 1 H, H-1), 4.72 (d, J = 12.5 Hz, 1 H, CH2O), 4.59 (d,
2.12, 2.04, 1.98 (4 s, 36 H, MeCO) ppm. 13C NMR (CDCl3, J = 12.5 Hz, 1 H, CH2O), 4.25 (dd, J = 11.9, 4.1 Hz, 1 H, H-6),
75 MHz): δ = 170.6, 170.0, 169.9, 169.67 (CO), 160.8 (Ar), 123.5
(C-5 triazole), 119.8 (Ar), 96.8 (C-1), 69.4, 69.0, 68.7, 66.0 (C-
4.05 (m, 2 H, H-5,6), 2.13, 2.10, 2.05, 1.98 (4 s, 12 H, 4 Ac) ppm.
13C NMR (CDCl3, 75 MHz): δ = 170.8, 170.1, 170.0, 169.7, 144.2,
2,3,4,5), 63.1 (OMe), 62.3 (C-6), 60.3 (CH2O), 43.9 (CH2N), 21.0, 137.5, 123.7, 96.9, 69.5, 69.0, 68.7, 66.1, 62.3, 60.7, 48.2, 20.9, 20.8,
20.8, 20.7, 20.6 (MeCO) ppm. HRMS (FAB+): calcd. for
20.7, 20.6 ppm. HRMS (FAB+): calcd. for C114H144N18O60 [M +
C63H81N9O33 [M + Na]+ 1514.48; found 1514.40.
Na]+ 2748.870; found 2748.860.
2448
www.eurjoc.org
© 2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Eur. J. Org. Chem. 2009, 2441–2453

Click Multivalent Homogeneous Neoglycoconjugates
1,2,3,4,5,6-Hexakis{4-(2,3,4,6-tetra-O-acetyl-α- -mannopyranosyl-
D
Methyl 2,3,4,6-Tetrakis-O-{4-(2,3,4,6-tetra-O-acetyl-α-
D-manno-
thiomethyl)-1H-1,2,3-triazol-1-ylmethyl}benzene (5Ib): Obtained
from 1b (722 mg) and 3I (102 mg) following the general procedure
(Method C). Column chromatography Cl2CH2/MeOH (25:1) gave
5Ib as a solid (457 mg, 65%); m.p. 186–188 °C. [α]2D0 = +163 (c = 1
pyranosyloxyethyl)-1H-1,2,3-triazol-1-yl}-α- -glucopyranoside
D
(6Kd): Obtained from 2d (500 mg) and 4K (87 mg) following the
general procedure (Method D). Column chromatography (EtOAc/
MeOH, 15:1) gave 6Kd as a solid (442 mg, 88%); m.p. 95 °C (dec).
in chloroform); IR (KBr): ν = 1749, 1371, 1229, 1050 cm–1. 1H [α]2D0 = +46 (c = 1 in chloroform); IR (KBr): ν = 1749, 1371, 1226,
˜
˜
NMR (CDCl3, 300 MHz): δ = 7.34 (s, 6 H), 5.82 (s, 12 H), 5.32 (t, 1138, 1094, 1046 cm–1. H NMR (CDCl3, 500 MHz): δ = 8.05 (s,
1
J = 9.5 Hz, 6 H), 5.23–5.17 (m, 18 H), 4.30 (m, 6 H), 4.26 (dd, J
= 12.1, 4.6 Hz, 6 H), 4.09 (dd, J = 11.8, 1.8 Hz, 6 H), 3.85 (d, J =
14.8 Hz, 6 H), 3.74 (d, J = 14.8 Hz, 6 H), 2.14, 2.10, 2.08, 1.98 (4
1 H), 7.93 (s, 2 H), 7.81 (s, 1 H), 5.27–5.16 (m, 12 H), 5.01–4.74
(several m, 11 H), 4.80–4.68 (m, 10 H), 4.24–3.59 (several m, 24
H), 3.39 (s, 3 H), 2.14, 2.13, 2.12, 2.09, 2.04, 2.03, 2.02, 1.98, 1.96,
s, 72 H) ppm. 13C NMR (CDCl3, 75 MHz): δ = 170.8, 170.1. 169.9, 1.96 (10 s, 48 H, 16 Ac) ppm. 13C NMR (CDCl3, 125 MHz): δ =
169.7, 145.0, 137.4, 122.7, 81.7, 70.9, 69.5, 66.2, 62.3, 48.1, 24.7,
21.0, 20.8, 20.7, 20.6 ppm. HRMS (FAB+): calcd. for
C
170.7, 170.0, 169.9, 169.8, 169.7, 144.9, 124.6, 97.7, 97.6, 81.4, 79.8,
70.0, 79.2, 68.9, 68.9, 68.6, 66.3, 65.7, 64.5, 64.3, 62.2, 55.2, 49.6,
20.9, 20.8, 20.7 ppm. HRMS (FAB+): calcd. for C83H114N12O46
[M + Na]+ 2037.68; found 2037.30.
114H144N18O54S6 [M + Na]+ 2844.733; found 2844.735.
Tetrakis-O-{4-(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyl)-1H-
1,2,3-triazol-1-ylmethyl}pentaerythritol (6Jc): Obtained from 2c
(447 mg) and 4J (72 mg) following the general procedure (Method
D). Column chromatography (EtOAc/hexane, 5:1 Ǟ 10:1) gave 6Jc
as a solid (603 mg, 79%); m.p. 104–106 °C (dec). [α]2D0 = +46 (c =
4-(2,3,4,6-Tetra-O-acetyl-α-
ylmethyl 2,3,4,6-Tetrakis-O-{4-(2,3,4,6-tetra-O-acetyl-α-
pyranosyl)-1H-1,2,3-triazol-1-yl}-β- -glucopyranoside (6Lc): Ob-
D
-mannopyranosyl)-1H-1,2,3-triazol-1-
D-manno-
D
tained from 2c (560 mg) and 4L (92 mg) following the general pro-
cedure (Method D). Column chromatography [EtOAc/hexane (5:1)
Ǟ EtOAc] gave 6Lc as a solid (420 mg, 75%); m.p. Ͼ 120 °C (dec).
1 in chloroform); IR (KBr): ν = 1750, 1370, 1225, 1122, 1041 cm–1.
˜
1H NMR (CDCl3, 300 MHz): δ = 7.85 (s, 4 H, H-5 triazole), 6.11
(d, J = 2.2 Hz, 4 H, H-1), 6.00–5.90 (m, 8 H, H-2,3), 5.39 (t, J =
9.0 Hz, 4 H, H-4), 4.60 (s, 8 H, CH2O), 4.36 (dd, J = 12.4, 5.2 Hz,
4 H, H-6), 4.08 (dd, J = 12.4, 2.4 Hz, 4 H, H-6), 3.92 (m, 4 H, H-
5), 3.50 (s, 8 H, CH2O), 2.18, 2.08, 2.04, 2.00 (4 s, 48 H, 16 Ac)
ppm. 13C NMR (CDCl3, 75 MHz): δ = 170.4, 169.5, 169.3 (CO),
145.7 (C-4 triazole), 123.0 (C-5 triazole), 83.6 (C-1), 71.9, 68.8,
68.2, 65.9 (C-2,3,4,5), 68.9, 64.5, 61.5 (CH2O), 49.4 (CH2N), 45.4
[C(CH2)4], 20.6, 20.5, 20.4 (MeCO) ppm. HRMS (FAB+): calcd.
for C73H96N12O40 [M + Na]+ 1803.57; found 1803.78.
[α]2D0 = +52 (c = 1 in chloroform); IR (KBr): ν = 1750, 1370, 1224,
˜
1122, 1040 cm–1. 1H NMR (CDCl3, 300 MHz): δ = 8.16, 8.12, 8.09,
7.99, 7.84, (5 s, 5 H, H-5 triazole), 6–08 (s, 3 H, H-1 Man), 6.05
(d, J = 1.9 Hz, 1 H, H-1 Man), 6.01 (d, J = 1.8 Hz, 1 H, H-1 Man),
5.97–5.82 (m, 10 H, H-2,3 Man), 5.39–5.30 (m, 5 H, H-4 Man),
5.03 (d, J = 10.7 Hz, 1 H), 4.99 (d, J = 11.7 Hz, 1 H), 4.97 (d, J =
11.9 Hz, 1 H), 4.94 (d, J = 11.0 Hz, 1 H), 4.90 (d, J = 11.2 Hz, 1
H), 4.83 (d, J = 10.7 Hz, 1 H), 4.80 (d, J = 11.3 Hz, 1 H), 4.74 (d,
J = 12.5 Hz, 1 H), 4.68 (d, J = 12.4 Hz, 1 H), 4.66 (d, J = 11.3 Hz,
1 H, 5 CH2O), 4.42 (d, J = 10.7 Hz, 1 H, H-1 Glc), 4.35–4.23 (m,
6 H, H-6 Glc, H-6 Man), 4.02 (br. d, J = 12.4 Hz, 5 H, H-6 Man),
3.93–3.80 (m, 6 H, H-5 Man, H-6 Glc), 3.53–3.33 (m, 4 H, H-
2,3,4,5 Glc), 2.17, 2.08, 2.07, 2.06, 2.04, 2.02 (6 s, 60 H, 20 Ac)
ppm. 13C NMR (CDCl3, 75 MHz): δ = 170.4, 169.6, 169.5, 169.3
(CO), 145.5, 145.2, 145.1, 144.9 (C-4 triazole), 124.5, 124.2, 123.8,
123.3 (C-5 triazole), 102.5 (C-1 Glc), 83.9, 83.8, 83.6 (C-1 Man),
81.8, 77.1, 74.5, 72.1 (C-2,3,4,5 Glc), 72.1, 72.0, 71.8 (C-5 Man),
69.0 (C-6 Glc), 68.9, 68.8, 68.2, 68.1 (C-2,3 Man), 65.9, 65.8 (C-4
Man), 65.3, 65.2, 64.4, 62.7 (5 CH2O), 61.5 (C-6 Man), 20.7, 20.5
(MeCO) ppm. HRMS (FAB+): calcd. for C91H117N15O51 [M +
Na]+ 2258.692; found 2258.761.
Tetrakis-O-{4-(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyloxyethyl)-
1H-1,2,3-triazol-1-ylmethyl}pentaerythritol (6Jd): Obtained from
2d (500 mg) and 4J (72 mg) following the general procedure
(Method D). Column chromatography (EtOAc) gave 6Jd as a solid
(390 mg, 80%); m.p. 80–82 °C (dec). [α]2D0 = +32 (c = 1, chloro-
1
form); IR (KBr): ν = 1744, 1369, 1224, 1136, 1086, 1044 cm–1. H
˜
NMR (CDCl3, 300 MHz): δ = 7.77 (s, 4 H, H-5 triazole), 5.30–
5.16 (m, 12 H, H-2,3,4), 4.84 (s, 4 H, H-1), 4.63–4.60 (m, 8 H,
CH2N), 4.22 (dd, J = 12.4, 5.0 Hz, 4 H, H-6), 4.12 (m, 4 H,
NCH2CH2O), 4.05 (br. d, J = 11.0 Hz, 4 H, H-6), 3.93 (m, 4 H,
NCH2CH2O), 3.62 (m, 4 H, H-5), 3.50 (s, 8 H, CH2O), 2.14, 2.09,
2.04, 1.99 (4 s, 48 H, 16 Ac) ppm. 13C NMR (CDCl3, 75 MHz): δ
= 170.4, 169.7, 169.5 (CO), 145.3 (C-4 triazole), 123.7 (C-5 tri-
azole), 97.3 (C-1), 67.1, 68.9, 65.5 (C-2,3,4,5), 68.7, 66.1 64.5, 62.0
(CH2O), 49.4 (CH2N), 45.1 [C(CH2)4], 20.6, 20.5, 20.4 (MeCO)
ppm. HRMS (FAB+): calcd. for C81H112N12O44 [M + Na]+
1979.68; found 1979.80.
4-(2,3,4,6-Tetra-O-acetyl-α-
azol-1-ylmethyl 2,3,4,6-Tetrakis-O-{4-(2,3,4,6-tetra-O-acetyl-α-
mannopyranosyloxyethyl)-1H-1,2,3-triazol-1-yl}-β- -glucopyran-
D-mannopyranosyloxyethyl)-1H-1,2,3-tri-
D
-
D
oside (6Ld): Obtained from 2d (625 mg) and 4L (92 mg) following
the general procedure (Method D). Column chromatography
(EtOAc/MeOH, 10:1) gave 6Ld as a solid (540 MG, 88%); m.p.
Methyl 2,3,4,6-Tetrakis-O-{4-(2,3,4,6-tetra-O-acetyl-α-
D-manno-
pyranosyl)-1H-1,2,3-triazol-1-yl}-α- -glucopyranoside (6Kc): Ob-
D
93 °C (dec). [α]2D0 = +18 (c = 1 in chloroform); IR (KBr): ν = 1748,
˜
tained from 2c (447 mg) and 4K (87 mg) following the general pro-
cedure (Method D). Column chromatography (EtOAc Ǟ EtOAc/
MeOH, 15:1) gave 6Kc as a solid (367 mg, 80%); m.p. 105 °C (dec).
1370, 1256, 1138, 1046 cm–1. 1H NMR (CDCl3, 500 MHz): δ =
8.07 (s, 1 H, H-5 triazole), 8.01 (s, 1 H, H-5 triazole), 7.90 (s, 1 H,
H-5 triazole), 7.80 (s, 1 H, H-5, triazole), 5.20 (m, 15 H), 5.00–4.76
(m, 12 H), 4.68–4.57 (m, 12 H), 4.47 (d, J = 7.2 Hz, 1 H), 4.20 (m,
5 H), 4.14 (m, 5 H), 4.04 (m, 5 H), 3.93 (m, 5 H), 3,80–3.38 (several
m, 12 H), 2.11, 2.07, 2.02, 2.07, 2.01, 2.00, 1.96, 1.95 (7 s, 60 H)
ppm. 13C NMR (CDCl3, 125 MHz): δ = 170.6, 169.9, 169.9, 169.9,
169.7, 145.3 (C-4 triazole), 145.0, 144.9, 124.7, 124.5, 124.3, 102.6,
97.8, 97.7, 82.1, 81.9, 74.4, 69.3, 69.0, 68.9, 66.4, 65.8, 64.6, 62.3,
49.5, 21.1, 20.9, 20.8, 20.7 ppm. HRMS (FAB+): calcd. for
C101H137N15O56 [M + Na]+ 2478.82; found 2478.30.
[α]2D0 = +58 (c = 0.5 in chloroform); IR (KBr): ν = 1751, 1371,
˜
1
1269, 1042 cm–1. H NMR (CDCl3, 500 MHz): δ = 8.16 (s, 1 H),
8.06 (s, 1 H), 8.04 (s, 1 H), 7.83 (s, 1 H), 6.11–5.88 (m, 12 H), 5.40
(m, 4 H), 5.04–4.58 (m, 9 H), 4.34 (m, 4 H), 4.10–3.58 (several m,
14 H), 3.39 (s, 3 H), 2.17, 2.10, 2.08, 2.07, 2.06, 2.05, 2.04, 2.03,
2.02, 2.01 (10 s, 48 H) ppm. 13C NMR (CDCl3, 75 MHz): δ =
170.6, 169.7, 169.7, 169.6, 169.4, 145.5, 145.3, 145.2, 124.3, 124.2,
123.4, 97.8, 84.0, 83.8, 81.4, 80.0, 77.4, 72.1, 72.0, 71.9, 70.0, 69.1,
68.9, 68.4, 68.2, 66.2, 66.1, 66.0, 65.5, 64.5, 61.6, 55.3, 20.8, 20.6
ppm. HRMS (FAB+): calcd. for C75H98N12O42 [M + Na]+ 1861.58;
found 1861.71.
1,2,3,4,5,6-Hexakis-O-{4-(2,3,4,6-tetra-O-acetyl-α-
D-mannopyran-
osyl)-1H-1,2,3-triazol-1-ylmethyl}- -mannitol (6Mc): Obtained
D
Eur. J. Org. Chem. 2009, 2441–2453
© 2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.eurjoc.org
2449

F. Santoyo-Gonzalez et al.
FULL PAPER
from 2c (670 mg) and 4M (102 mg) following the general procedure
(Method D). Column chromatography [EtOAc/hexane (50:1) Ǟ
EtOAc/MeOH (15:1)] gave 6Mc as a foamy solid (442 mg, 67%):
75%); m.p. 111–113 °C. [α]2D0 = +63 (c = 1 in H O); IR (KBr): ν =
˜
2
3433, 1132, 1058 cm–1. H NMR ([D6]DMSO, 300 MHz, selected
1
signals): δ = 8.16 (s, 2 H), 7.35 (br. t, J = 4.4 Hz, 2 H), 7.14 (br. t,
J = 4.4 Hz, 2 H), 5.82 (s, 4 H), 4.73 (s, 2 H), 4.68 (d, J = 12.3 Hz,
[α]2D0 = +55 (c = 1 in chloroform); IR (KBr): ν = 3143, 2936, 1751,
˜
1628, 1434, 1369, 1225, 1122, 1040 cm–1 1H NMR (CDCl3, 2 H), 4.52 (d, J = 12.2 Hz, 2 H), 3.67 (d, J = 11.3 Hz, 2 H), 3.58
.
300 MHz): δ = 7.99, 7.97, 7.95, (3 s, 6 H), 6.11–6.08 (m, 6 H), 5.98– (s, 2 H) ppm. 13C NMR ([D6]DMSO, 75 MHz): δ = 144.0, 134.2,
5.88 (m, 12 H), 5.43–5.35 (m, 6 H), 4.80–3.97 (several m, 38 H), 129.2, 129.0, 124.5, 99.1, 74.2, 71.0, 70.2, 67.1, 61.3, 59.1, 50.0 ppm.
2.16, 2.15, 2.07, 2.05, 2.00 (5 s, 72 H) ppm. 13C NMR (CDCl3, HRMS (FAB+): calcd. for C26H36N6O12 [M + Na]+ 647.2289;
75 MHz): δ = 170.5, 169.7, 169.5, 145.3, 124.3, 124.0, 83.9, 78.3,
71.9, 69.2, 68.3, 68.2, 69.5, 61.7, 20.7, 20.5 ppm. HRMS (FAB+):
calcd. for C108H140N18O60 [M + Na]+ 2673.83; found 2673.73.
found 647.2282.
1,2-Bis{4-(α- -mannopyranosylthiomethyl)-1H-1,2,3-triazol-1-
D
ylmethyl}benzene (7Bf): Obtained from 5Bb (496 mg) following the
general procedure (Method A). Column chromatography (EtOAc/
MeOH, 1:1 Ǟ 1:3) and lyophilization gave 7Bf (234 mg, 100%) as
a solid; m.p. 150–152 °C (dec). [α]2D0 = +133 (c = 1 in H2O); IR
1,2,3,4,5,6-Hexakis-O-{4-(2,3,4,6-tetra-O-acetyl-α-
D-mannopyran-
osyloxyethyl)-1H-1,2,3-triazol-1-ylmethyl}- -mannitol (6Md): Ob-
D
tained from 2d (750 mg) and 4M (102 mg) following the general
procedure (Method D). Column chromatography (EtOAc/MeOH,
10:1) gave 6Md as a foamy solid (537 mg, 74%): [α]2D0 = +30 (c =
(KBr): ν = 3421, 2929, 1071 cm–1 1H NMR ([D6]DMSO,
.
˜
300 MHz, selected signals): δ = 7.85 (s, 2 H), 7.45–7.35 (m, 2 H),
7.35–7.20 (m, 2 H), 5.76 (s, 4 H), 3.85 (d, J = 14.4 Hz, 2 H), 3.76
(d, J = 14.4 Hz, 2 H) ppm. 13C NMR ([D6]DMSO, 75 MHz): δ =
146.4, 135.0, 131.4, 130.7, 124.8, 85.3, 75.2, 73.2, 73.2, 68.9, 62.7,
52.0, 24.7 ppm. HRMS (FAB+): calcd. for C26H36N6O10S2 [M +
Na]+ 679.1832; found 679.1835.
1 in chloroform); IR (KBr): ν = 1745, 1371, 1230, 1139, 1048, 980
˜
cm–1. 1H NMR (CDCl3, 300 MHz): δ = 7.91 (s, 2 H), 7.90 (s, 2 H),
7.85 (s, 2 H), 5.28–5.22 (m, 18 H), 4.85 (m, 3 H), 4.83 (s, 3 H),
4.77–4.61 (m, 24 H), 4.22 (dd, J = 12.3, 4.7 Hz, 6 H); 4.13–4.70
(several m, 32 H), 2.13, 2.08, 2.04, 1.97 (4 s, 72 H) ppm. 13C NMR
(CDCl3, 75 MHz): δ = 170.5, 169.8, 169.6, 145.0, 144.8, 124.3, 97.5,
78.4, 78.2, 69.1, 68.8, 68.8, 66.1, 65.6, 65.2, 64.2, 62.7, 62.1, 49.5,
49. 4, 20.7, 20.6,20.5 ppm. HRMS (FAB+): calcd. for
1,3-Bis{(-α-D-mannopyranosyloxymethyl)-1H-1,2,3-triazol-1-
ylmethyl}benzene (7Ce): Obtained from 5Ca (480 mg) following the
general procedure (Method A). Column chromatography (EtOAc/
MeOH, 1:1 Ǟ 1:2) and lyophilization gave 7Ce (244 mg, 78%) as
C
120H164N18O66 [M + Na]+ 2935.99; found 2935.90.
a deliquescent solid: [α]2D0 = +54 (c = 1 in H O); IR (KBr): ν =
˜
2
General Procedure for the Zemplen Deacetylation of Peracetylated
Neoglycoconjugates. Method A: A solution of the corresponding
per-O-acetylated neoglycoconjugate (0.1 mmol) in MeOH (8 mL)
containing Et3N (0.8 mL) was refluxed for 2–3 h. The evaporation
of the solvent was followed by purification of the crude material
by short-column flash chromatography.
3393, 2930, 1131, 1058 cm–1. 1H NMR ([D6]DMSO, 300 MHz, se-
lected signals): δ = 8.16 (s, 2 H), 7.38 (t, J = 7.5 Hz, 1 H), 7.34 (s,
1 H), 7.25 (d, J = 7.4 Hz, 2 H), 5.58 (s, 4 H), 4.71 (s, 2 H), 4.66 (d,
J = 12.3 Hz, 2 H), 4.50 (d, J = 12.2 Hz, 2 H), 3.67 (d, J = 11.2 Hz,
2 H), 3.57 (s, 2 H) ppm. 13C NMR ([D6]DMSO, 75 MHz): δ =
143.9, 136.6, 129.3, 127.8, 127.6, 124.3, 99.1, 74.2, 70.9, 70.2, 67.0,
61.3, 59.1, 52.6 ppm. HRMS (FAB+): calcd. for C26H36N6O12 [M
+ Na]+ 647.2289; found 647.2284.
Method B: To a solution of the corresponding per-O-acetylated
neoglycoconjugate (0.1 mmol) in MeOH (15 mL) was added Na-
OMe (1 , 0.1 mL). After the solution was maintained overnight
at room temp., the solvent was evaporated, and the resulting crude
material was purified by short-column flash chromatography.
1,3-Bis{(-α-D-mannopyranosylthiomethyl)-1H-1,2,3-triazol-1-
ylmethyl}benzene (7Cf): Obtained from 5Cb (496 mg) following the
general procedure (Method A). Column chromatography (EtOAc/
MeOH, 1:1 Ǟ 1:2) and lyophilization gave 7Cf (263 mg, 80%) as
1,2-Bis{4-(α-D-mannopyranosyloxymethyl)-1H-1,2,3-triazol-1-
yl}ethane (7Ae): Obtained from 5Aa (442 mg) following the general
a deliquescent solid: [α]2D0 = +165 (c = 1 in H O); IR (KBr): ν =
˜
2
3409, 2929, 1071 cm–1. H NMR ([D6]DMSO, 300 MHz, selected
1
procedure (Method A). Column chromatography (EtOAc/MeOH,
1:2) gave 7Ae (250 mg, 91%) as a syrup: [α]2D0 = +60 (c = 1 in H2O);
signals): δ = 8.06 (s, 2 H), 7.37 (t, J = 7.6 Hz, 1 H), 7.29 (s, 1 H),
7.13 (d, J = 7.4 Hz, 2 H), 5.56 (s, 4 H), 3.84 (d, J = 14.4 Hz, 2 H),
3.75 (d, J = 14.4 Hz, 2 H) ppm. 13C NMR ([D6]DMSO, 75 MHz):
δ = 144.5, 136.7, 129.3, 127.6, 127.4, 123.4, 84.0, 74.8, 71.7, 71.5,
67.3, 61.1, 52.5, 23.5 ppm. HRMS (FAB+): calcd. for
C26H36N6O10S2 [M + Na]+ 679.1832; found 679.1830.
1
IR (KBr): ν = 3392, 1655, 1411, 1131, 1056 cm–1. H NMR ([D ]-
˜
6
DMSO, 300 MHz): δ = 8.02 (s, 2 H), 4.90 (s, 4 H), 4.86–4.80 (br.
s, 8 H), 4.68 (s, 2 H), 4.62 (d, J = 12.3 Hz, 2 H), 4.48 (d, J =
12.3 Hz, 2 H), 3.7–3.3 (several m, 12 H) ppm. 13C NMR ([D6]-
DMSO, 75 MHz): δ = 145.6, 125.9, 100.6, 75.0, 72.4, 71.9, 68.6,
63.0, 60.4, 50.9 ppm. HRMS (FAB+): calcd. for C20H32N6O12 [M
+ Na]+ 571.1976; found 571.1980.
1,4-Bis{(-α-D-mannopyranosyloxymethyl)-1H-1,2,3-triazol-1-
ylmethyl}benzene (7De): Obtained from 5Da (480 mg) following the
1,2-Bis{4-(α-D-mannopyranosylthiomethyl)-1H-1,2,3-triazol-1-
general procedure (Method A). Column chromatography (EtOAc/
MeOH, 1:1) gave 7De as a foamy solid (312 mg, 100%): [α]2D0
=
yl}ethane (7Af): Obtained from 5Aa (458 mg) following the general
procedure (Method A). Column chromatography (EtOAc/MeOH,
1:3) gave 7Af (229 mg, 79%) as a syrup: [α]2D0 = +110 (c = 0.5 in
DMSO). 1H NMR ([D6]DMSO, 300 MHz, selected signals): δ =
7.78 (s, 2 H), 5.08 (s, 4 H), 4.84 (s, 6 H), 3.81 (d, J = 14.4 Hz, 2 H),
3.72 (d, J = 14.4 Hz, 2 H) ppm. 13C NMR ([D6]DMSO, 75 MHz): δ
= 146.1, 125.1, 84.8, 75.1, 73.1, 73.1, 68.8, 62.7, 50.9, 24.4 ppm.
HRMS (FAB+): calcd. for C20H32N6O10S2 [M + Na]+ 603.1519;
found 603.1523.
+44 (c = 1 in H O); IR (KBr): ν = 3372, 2926, 1566, 1411, 1130,
˜
2
1052, 1024 cm–1. 1H NMR ([D6]DMSO, 300 MHz, selected sig-
nals): δ = 8.15 (s, 2 H), 7.32 (s, 4 H), 5.57 (s, 4 H), 4.70 (s, 2 H),
4.65 (d, J = 12.2 Hz, 2 H), 4.49 (d, J = 12.2 Hz, 2 H), 3.67 (d, J =
11.0 Hz, 2 H), 3.56 (s, 2 H) ppm. 13C NMR ([D6]DMSO, 75 MHz):
δ = 143.9, 136.0, 128.5, 124.2, 99.1, 74.2, 70.9, 70.2, 67.0, 62.3,
59.1, 52.5 ppm. HRMS (FAB+): calcd. for C26H36N6O12 [M +
Na]+ 647.2289; found 647.2286.
1,2-Bis{4-(α-D-mannopyranosyloxymethyl)-1H-1,2,3-triazol-1- 1,4-Bis{(-α-D-mannopyranosylthiomethyl)-1H-1,2,3-triazol-1-
ylmethyl}benzene (7Be): Obtained from 5Ba (480 mg) following the
ylmethyl}benzene (7Df): Obtained from 5Db (496 mg) following the
general procedure (Method A). Column chromatography (EtOAc/
general procedure (Method A). Column chromatography (EtOAc/
MeOH, 2:1 Ǟ 1:1) and lyophilization gave 7Be as a solid (234 mg, MeOH, 1:1) and lyophilization gave 7Df (233 mg, 71%) as a foamy
2450
www.eurjoc.org
© 2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Eur. J. Org. Chem. 2009, 2441–2453

Click Multivalent Homogeneous Neoglycoconjugates
solid: [α]2D0 = +61 (c = 0.5 in H O); IR (KBr): ν = 3402, 1647, 1438
1,3,5-Tris{4-(α-D-mannopyranosyloxymethyl)-1H-1,2,3-triazol-1-
˜
2
cm–1. H NMR ([D6]DMSO, 300 MHz): δ = 8.03 (s, 2 H), 7.29 (s,
ylmethyl}-2,4,6-trimethoxybenzene (7Ge): Obtained from 5Ga
(372 mg) following the general procedure (Method A). Column
chromatography (CH3CN/H2O, 5:1 Ǟ 3:1) and lyophilization gave
7Ge (187 mg, 76%) as a foamy solid: [α]2D0 = +57.9 (c = 0.19,
1
4 H), 5.53 (s, 4 H), 5.11 (s, 2 H), 9.96 (d, J = 4.3 Hz, 2 H), 4.73 (d,
J = 4.5 Hz, 2 H), 4.66 (d, J = 4.5 Hz, 2 H), 4.52 (t, J = 5.8 Hz, 2
H), 3.85 (d, J = 14.3 Hz, 2 H), 3.73 (d, J = 14.5 Hz, 2 H), 3.65–
3.60 (m, 6 H), 3.55–3.35 (m, 6 H) ppm. 13C NMR ([D6]DMSO,
75 MHz): δ = 144.6, 136.0, 128.4, 83.9, 74.8, 71.6, 71.5, 67.3, 61.1,
52.4, 23.4 ppm. HRMS (FAB+): calcd. for C26H36N6O10S2 [M +
Na]+ 679.1832; found 679.1833.
DMSO); IR (KBr): ν = 3394, 2920, 2866, 1616, 1587, 1454, 1414,
˜
1
1344, 1101, 1069 cm–1. H NMR ([D6]DMSO + D2O, 300 MHz):
δ = 8.04 (s, 3 H), 5.53 (s, 6 H, CH2N), 4.68 (s, 3 H, H-1), 4.62 (d,
J = 12.1 Hz, 3 H, CH2O), 4.49 (d, J = 12.1 Hz, 3 H, CH2O), 3.66
(s, 3 H, OMe), 3.64 (m, 3 H, H-6), 3.54 (br. s, 3 H, H-2), 3.50–3.20
(m, 12 H, H-3,4,5,6) ppm. 13C NMR ([D6]DMSO, 75 MHz): δ =
159.9, 143.4, 124.6, 119.6, 99.0, 74.1, 70.9, 70.2, 67.0, 62.3, 61.3,
59.0, 43.3 ppm. HRMS (MALDI-TOF): calcd. for C39H57N9O21
[M + Na]+ 1010.35; found 1010.29.
1,3,5-Tris{2-[4-(α-D-mannopyranosyloxymethyl)-1H-[1,2,3]triazol-1-
yl]-ethoxy}benzene (7Ee): Obtained from 5Ea (372 mg) following
the general procedure (Method A). Column chromatography
(CH3CN/H2O, 4:1) and lyophilization gave 7Ee (210 mg, 85%) as
a foamy solid: [α]2D0 = +52 (c = 0.5 in DMSO); IR (KBr): ν = 3400,
˜
1171, 1129, 1064 cm–1. 1H NMR ([D6]DMSO, 300 MHz): δ = 8.12 1,3,5-Tris{4-(α-
D
-mannopyranosylthiomethyl)-1H-1,2,3-triazol-1-
(s, 3 H, H-5 triazole), 6.11 (s, 6 H, ArH), 4.71 (s, 3 H, H-1), 4.69 ylmethyl}-2,4,6-trimethoxybenzene (7Gf): Obtained from 5Gb
(m, 18 H, 12 CH2N, 6 OH), 4.65 (d, J = 12.3 Hz, 3 H, CH2O- (384 mg) following the general procedure (Method A). Column
Man), 4.50 (d, J = 12.0 Hz, 3 H, CH2O-Man), 4.50 (br. s, 3 H,
OH), 4.45 (t, J = 5.8 Hz, 3 H, OH), 4.34 (t, J = 5.0 Hz, 2 H,
CH2O), 3.67 (dd, J = 11.3, 6.1 Hz, 3 H, H-6), 3.55 (br. s, 1 H, H-
chromatography (CH3CN/H2O, 5:1 Ǟ 3:1) and lyophilization gave
7Ge (214 mg, 83%) as a foamy solid: [α]2D0 = +210 (c = 0.5, in
DMSO); IR (KBr): ν = 3384, 1200, 1101, 1070 cm–1. 1H NMR
˜
2), 3.50–3.32 (m, 12 H, H-3,4,5,6) ppm. 13C NMR ([D6]DMSO, ([D6]DMSO, 300 MHz): δ = 7.95 (s, 3 H, H-5 triazole), 5.51 (s, 6
75 MHz): δ = 159.5, 143.4, 124.4, 98.9, 94.5, 74.0, 70.8, 70.1, 66.9,
66.1, 61.2, 58.9, 48.8 ppm. HRMS (FAB+): calcd. for C39H57N9O11
[M + Na]+ 1010.35; found 1010.40.
H, CH2N), 5.12 (s, 3 H), 5.00 (d, J = 3.9 Hz, 3 H), 4.82 (s, 3 H),
4.70 (s, 3 H), 4.55 (m, 3 H), 3.82 (d, J = 14.3 Hz, 3 H, CH2S), 3.74
(d, J = 14.3 Hz, 3 H, CH2S), 3.65 (s, 9 H, OMe), 3.65 (m, 9 H),
3.51–3.40 (m, 9 H) ppm. 13C NMR ([D6]DMSO, 75 MHz): δ =
159.7, 144.0, 123.4, 119.5, 83.8, 74.6, 71.5, 71.3, 67.2, 62.2, 61.0,
43.2, 23.3 ppm. HRMS (MALDI-TOF): calcd. for C39H57N9O18S3
[M + Na]+ 1058.29; found 1058.30.
1,3,5-Tris{2-[4-(α-D-mannopyranosyloxymethyl)-1H-[1,2,3]triazol-1-
yl]-ethoxy}benzene (7Ef): Obtained from 5Eb (360 mg) following
the general procedure (Method A). Column chromatography
(CH3CN/H2O, 4:1) and lyophilization gave 7Ef (209 mg, 81%) as
a solid; m.p. 117–119 °C. [α]2D0 = +177.4 (c = 0.5, DMSO); IR
1,2,4,5-Tetrakis{4-(α-D-mannopyranosyloxymethyl)-1H-1,2,3-tri-
1
(KBr): ν = 3392, 1170, 1067 cm–1. H NMR ([D ]DMSO + D O, azol-1-ylmethyl}benzene (7He): Obtained from 5Ha (460 mg) fol-
˜
6
2
300 MHz): δ = 8.04 (s, 3 H, H-5 triazole), 6.11 (s, 3 H, ArH), 5.14
(s, 3 H, H-1), 4.70 (br. s, 6 H, CH2N), 4.33 (t, J = 4.4 Hz, 6 H,
lowing the general procedure (Method A). Column chromatog-
raphy (EtOAc/MeOH, 1:3) and lyophilization 7He (258 mg, 95%)
CH2O), 3.84 (d, J = 14.3 Hz, 3 H, CH2S), 3.76 (d, J = 14.3 Hz, 3 as a foamy solid: [α]2D0 = +58 (c = 1 in H O); IR (KBr): ν = 3365,
˜
2
H, CH2S), 3.71–3.65 (m, 9 H, H-2,3,6), 3.50–3.41 (m, 9 H, H-4,5,6) 1645, 1230, 1134, 1059 cm–1. H NMR ([D6]DMSO, 300 MHz): δ
1
ppm. 13C NMR ([D6]DMSO, 75 MHz): δ = 159.5, 144.1, 123.4,
94.5, 83.8, 74.6, 71.5, 71.3, 67.2, 66.1, 61.0, 48.8, 23.3 ppm. HRMS
(FAB+): calcd. for C39H57N9O18S3 [M + Na]+ 1058.29; found
1058.30.
= 8.08 (s, 4 H, H-5 triazole), 6.98 (s, 2 H), 5.73 (s, 8 H), 4.71 (s, 4
H), 4.64 (d, J = 12.3 Hz, 4 H), 4.50 (d, J = 12.2 Hz, 4 H), 4.65–
4.50 (m, 16 H), 3.70-3.33 (several m, 24 H) ppm. 13C NMR ([D6]-
DMSO, 75 MHz): δ = 143.9, 134.8, 124.4, 99.0, 74.1, 70.9, 70.2,
67.0, 61.3, 59.0, 49.0 ppm. HRMS (MALDI-TOF): calcd. for
C46H66N12O24 [M + Na]+ 1193.4210; found 1193.316.
1,3,5-Tris{4-(α-D-mannopyranosylmethyl)-1H-1,2,3-triazol-1-
ylmethyl}-2,4,6-trimethylbenzene (7Fe): Obtained from 5Fa
(360 mg) following the general procedure, (Method A). Column
chromatography (EtOAc/MeOH, 1:3) and lyophilization gave 7Fe
(213 mg, 91%) as a foamy solid: [α]2D0 = +49 (c = 1, H2O); IR
1,2,4,5-Tetrakis{4-(α-D-mannopyranosylthiomethyl)-1H-1,2,3-tri-
azol-1-ylmethyl}benzene (7Hf): Obtained from 5Hb (476 mg) fol-
lowing the general procedure (Method A). Column chromatog-
raphy (EtOAc/MeOH, 1:3) and lyophilization gave 7Hf (86%) as a
(KBr): ν = 3410, 1643, 1227, 1131, 1058 cm–1. 1H NMR ([D6]-
˜
DMSO, 300 MHz): δ = 7.96 (s, 3 H), 5.67 (s, 6 H), 4.77 (br. s, 12
H), 4.70 (s, 3 H), 4.62 (d, J = 12.2 Hz, 3 H), 4.47 (d, J = 12.2 Hz,
deliquescent solid: [α]2D0 = +249 (c = 0.5 in DMSO); IR (KBr): ν =
˜
3401, 2926, 1638, 1228, 1073 cm–1
.
1H NMR ([D6]DMSO,
3 H), 3.72–3.30 (m, 18 H), 2.41 (s, 9 H) ppm. 13C NMR ([D6]- 300 MHz, selected signals): δ = 7.78 (s, 4 H), 7.17 (s, 2 H), 5.74 (s,
DMSO, 75 MHz): δ = 143.4, 139.3, 130.8, 123.8, 99.1, 74.1, 70.9,
70.1, 67.0, 61.3, 59.0, 48.4, 16.4 ppm. HRMS (MALDI-TOF): 143.6, 131.8, 129.5, 121.6, 81.7, 70.6, 68.8, 68.6, 64.3, 58.0, 47.9,
8 H), 5.20 (s, 4 H) ppm. 13C NMR ([D6]DMSO, 75 MHz): δ =
calcd. for C39H57N9O18 [M + Na]+ 962.371; found 962.410.
21.3 ppm. HRMS (MALDI-TOF): calcd. for C46H66N12O20S4 [M
+ Na]+ 1257.330; found 1257.335.
1,3,5-Tris{4-(α- -mannopyranosylthiomethyl)-1H-1,2,3-triazol-1-
D
ylmethyl}-2,4,6-trimethylbenzene (7Ff): Obtained from 5Fb
(372 mg) following the general procedure (Method A). Column
chromatography (EtOAc/MeOH, 1:2) and lyophilization gave 7Ff
(212 mg, 91%) as a deliquescent solid: [α]2D0 = +154 (c = 1, H2O).
1H NMR ([D6]DMSO, 300 MHz, selected signals): δ = 7.85 (s, 3
H), 5.64 (s, 6 H), 5.13 (s, 3 H), 3.82 (d, J = 14.3 Hz, 3 H), 3.72 (d,
J = 14.4 Hz, 3 H) ppm. 13C NMR ([D6]DMSO, 75 MHz): δ =
144.1, 139.3, 130.9, 122.8, 84.0, 74.8, 71.6, 71.5, 67.3, 61.1, 56.0,
23.5, 16.4 ppm. HRMS (MALDI-TOF): calcd. for C39H57N9O15S3
[M + Na]+ 1010.303; found 1010.296.
1,2,3,4,5,6-Hexakis{4-(α-D-mannopyranosyloxymethyl)-1H-[1,2,3]-
1-methyl}benzene (7Ie): Obtained from 5Ia (688 mg) following the
general procedure (Method A). The white solid (7Ie) that appears
is filtered, washed with cold MeOH and dried (395 mg, 92%); m.p.
178–180 °C. [α]2D0 = +49 (c = 1 in H2O). 1H NMR ([D6]DMSO,
300 MHz, selected signals): δ = 7.78 (s, 6 H), 4.76 (s, 12 H) ppm.
13C NMR ([D6]DMSO, 100 MHz): δ = 143.3, 137.4, 124.1, 99.0,
74.0, 70.9, 70.1, 67.0, 61.3, 58.9, 47.7 ppm. HRMS (MALDI-
TOF): calcd. for C66H96N18O36 [M + Na]+ 1739.613; found
1739.52.
Eur. J. Org. Chem. 2009, 2441–2453
© 2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.eurjoc.org
2451

F. Santoyo-Gonzalez et al.
FULL PAPER
1,2,3,4,5,6-Hexakis{4-(α-
D
-mannopyranosyltjiomethyl)-1H-[1,2,3]-
pyranoside (8Lg): Obtained from 6Lc (558 mg) following the gene-
ral procedure (Method B). Column chromatography (EtOAc/
MeOH, 1:5) and lyophilization gave 8Lg as a foamy solid (325 mg,
1-methyl}benzene (7If): Obtained from 5Ib (705 mg) following the
general procedure (Method A). The white solid (7If) that appears
is filtered, washed with cold MeOH and dried (372 mg, 82%); m.p.
94%): [α]2D0 = +31 (c = 1 in H O); IR (KBr): ν = 3394, 1117, 1072
˜
2
Ͼ 230 °C (dec). [α]2D0 = +250 (c = 0.5 in DMSO); IR (KBr): ν =
cm–1. H NMR ([D6]DMSO, 500 MHz): δ = 8.27, 8.24, 8.23, 8.21,
1
˜
1
3392, 2929, 1073 cm–1. H NMR ([D6]DMSO, 300 MHz, selected
8.20 (5 s, 5 H), 5.89 (m, 5 H), 5.40–4.50 (several m, 29 H), 4.39 (m,
signals): δ = 7.67 (s, 6 H), 5.89 (s, 12 H), 5.11 (s, 6 H) ppm. 13C 5 H), 3.84 (m, 5 H), 3.60–3.10 (several m, 28 H) ppm. 13C NMR
NMR ([D6]DMSO, 75 MHz): δ = 143.9, 137.3, 123.1, 83.9, 74.6, ([D6]DMSO, 125 MHz): δ = 144.1, 143.9, 143.2, 123.8, 101.4, 85.7,
71.6, 71.4, 67.2, 61.1, 47.6, 23.4 ppm. HRMS (MALDI-TOF): 83.5, 81.2, 78.2, 78.1, 76.9, 73.8, 71.1, 68.2, 67.5, 64.8, 63.9, 61.7,
calcd. for C66H96N18O30S6 [M + Na]+ 1835.476; found 1835.476.
60.6, 57.5 ppm. HRMS (MALDI-TOF): calcd. for C51H77N15O31
[M + Na]+ 1418.40; found 1418.50.
Tetrakis-O-{4-(α- -mannopyranosyl)-1H-1,2,3-triazol-1-
D
ylmethyl}pentaerythritol (8Jg): Obtained from 6Jc (445 mg) follow-
ing the general procedure (Method B). Column chromatography
(EtOAc/MeOH, 1:5) and lyophilization gave 8Jg as a foamy solid
4-(α-
2,3,4,6-Tetrakis-O-{4-(α-
azol-1-yl}-β- -glucopyranoside (8Lh): Obtained from 6Ld (613 mg)
D
-Mannopyranosyloxyethyl)-1H-1,2,3-triazol-1-ylmethyl
D
-mannopyranosyloxyethyl)-1H-1,2,3-tri-
D
(236 mg, 85%): [α]2D0 = +34 (c = 1 in H O); IR (KBr): ν = 3410,
˜
following the general procedure (Method B). Column chromatog-
2
1658, 1384, 1085 cm–1. 1H NMR ([D6]DMSO, 300 MHz): δ = 8.18
(s, 4 H), 5.88 (d, J = 4.1 Hz, 4 H, H-1), 5.31 (br. s, 4 H), 5.12 (br.
s, 4 H), 5.04 (br. s, 4 H), 4.64 (br. s, 4 H), 4.46 (s, 8 H), 4.38 (br. s,
4 H), 3.85 (dd, J = 7.0, 3.2 Hz, 4 H), 3.60 (m, 12 H), 3.41 (m, 4
H), 3.36 (s, 8 H) ppm. 13C NMR ([D6]DMSO, 75 MHz): δ =
144.56, 124.1, 86.2, 78.8, 71.7, 69.5, 68.7, 68.2, 64.7, 61.2, 45.6 ppm.
HRMS (MALDI-TOF): calcd. for C41H64N12O24 [M + Na]+
1131.40; found 1131.50.
raphy (MeOH) and lyophilization gave 8Lh as a foamy solid
(350 mg, 87%): [α]2D0 = +31 (c = 1 in H O); IR (KBr): ν = 3392,
˜
2
1131, 1089, 1055 cm–1. H NMR ([D6]DMSO, 300 MHz, selected
1
signals): δ = 8.19 (s, 1 H), 8.16 (s, 1 H), 8.08 (s, 1 H), 8.05 (s, 2 H)
ppm. 13C NMR ([D6]DMSO, 75 MHz): δ = 144.2, 144.0, 125.9,
125.7, 125.5, 101.7, 99.7, 83.0, 80.8, 76.9, 73.5, 72.9, 70.6, 70.0,
67.9, 66.6, 65.6, 60.8, 50.2 ppm. HRMS (MALDI-TOF): calcd. for
C61H97N15O36 [M + Na]+ 1638.60; found 1638.50.
Tetrakis-O-{4-(α-D-mannopyranosyloxyethyl)-1H-1,2,3-triazol-1-
1,2,3,4,5,6-Hexakis-O-{4-(α-
D-mannopyranosyl)-1H-1,2,3-triazol-1-
ylmethyl}pentaerythritol (8Jh): Obtained from 6Jd (489 mg) follow-
ing the general procedure (Method B). Column chromatography
(EtOAc/MeOH, 1:5) and lyophilization gave 8Jh (280 mg) as a
ylmethyl}- -mannitol (8Mg): Obtained from 6Mc (662 mg) follow-
ing the general procedure (Method A). Column chromatography
D
(CH3CN/H2O, 2:1) gave 8Mg as a solid (370 mg, 90%); m.p. 203 °C
syrup (87%): [α]2D0 = +31 (c = 1 in H O); IR (KBr): ν = 3397, 1644,
20
(dec). [α]2D0 = +16 (c = 1 in H2O), [α] = +84 (c = 1 in H2O); IR
˜
2
436
1458, 1227, 1134, 1090, 1056 cm–1 1H NMR ([D6]DMSO,
.
(KBr): ν = 3411, 2924, 1640, 1437, 1114 cm–1. 1H NMR ([D6]-
˜
300 MHz): δ = 8.00 (s, 4 H), 4.61 (br. s, 4 H), 4.57 (s, 4 H), 4.57–
4.50 (m, 16 H), 4.43 (s, 8 H), 4.15 (br. s, 4 H), 3.91 (m, 4 H), 3.77
(m, 4 H), 3.62–3.33 (m, 24 H), 3.32 (s, 8 H) ppm. 13C NMR ([D6]-
DMSO, 75 MHz): δ = 144.0, 124.0, 99.8, 74.1, 70.8, 70.1, 68.7,
66.7, 64.9, 64.1, 61.1, 49.2, 44.9 ppm. HRMS (MALDI-TOF):
calcd. for C49H80N12O28 [M + Na]+ 1307.52; found 1307.54.
DMSO, 300 MHz, selected signals): δ = 8.28, 8.22, 8.21 (3 s, 6 H,
H-5 triazole), 5.93, 5.90, 5.89 (3 d, J = 3.7 Hz, 6 H, H-1 Man),
4.80–4.50 (several m, CH2O), 4.46–4.38 (m, 6 H, H-2 Man), 4.00–
3.50 (several m, 28 H, H-3,4,6,6 Man, CHO) ppm. 13C NMR ([D6]-
DMSO, 100 MHz): δ = 144.6, 144.5, 144.4 (C-4 triazole), 124.5,
124.3 (C-5 triazole), 86.3, 86.2 (C-1 Man), 78.7 (C-5 Man), 77.9,
71.7, 68.7, 68.1, 68.0 (C-2,3,4 Man), 64.2 (CH2O), 61.2 (C-6 Man),
49.1 ppm. HRMS (FAB+): calcd. for C60H92N18O36 [M + Na]+
1663.582; found 1663.621.
Methyl 2,3,4,6-Tetrakis-O-{4-(α-
D-mannopyranosyl)-1H-1,2,3-tri-
azol-1-yl}-α- -glucopyranoside (8Kg): Obtained from 6Kc (460 mg)
D
following the general procedure (Method B). Column chromatog-
raphy (MeOH) and lyophilization gave 8Kg as a foamy solid
(502 mg, 92%); m.p. 220 °C (dec). [α]2D0 = +73 (c = 1 in H2O); IR
1,2,3,4,5,6-Hexakis-O-{4-(α-D-mannopyranosyloxyethyl)-1H-1,2,3-
triazol-1-ylmethyl}- -mannitol (8Mh): Obtained from 6Md
D
(KBr): ν = 3366, 1644, 1435, 1114, 1044 cm–1. 1H NMR ([D6]-
˜
(485 mg) following the general procedure (Method A). Column
DMSO, 300 MHz, selected signals): δ = 8.28 (s, 1 H), 8.26 (s, 2 H),
8.21 (s, 1 H) 5.92 (br. s, 4 H) ppm. 13C NMR ([D6]DMSO,
75 MHz): δ = 144.2, 143.9, 143.8, 123.8, 123.7, 123.6, 97.1, 85.8,
85.7, 81.2, 79.2, 78.1, 77.1, 69.6, 68.8, 68.1, 67.6, 65.4, 64.9, 63.8,
63.4, 60.7, 54.4 ppm. HRMS (FAB+): calcd. for C43H66N12O26 [M
+ Na]+ 1189.411; found 1189.468.
chromatography (CH3CN/H2O, 2:1) gave 8Mh as a solid (283 mg,
20
90%); m.p. Ͼ 260 °C (dec). [α]2D0 = +12 (c = 1 in H2O), [α]
=
436
+49 (c = 1 in H O); IR (KBr): ν = 3417, 1652, 1139, 1102 cm–1.
˜
2
1H NMR ([D6]DMSO + D2O, 400 MHz): δ = 8.04, 7.98, 7.93 (3 s,
6 H, H-5 triazole), 4.70–4.40 (m, 18 H, CH2O-triazole, CH2N, H-
1 Man), 3.93, 3.80 (2 br. s, 16 H, CH2O, OCHO, CH2O-Man), 3.62
(m, 4 H, CH2O), 3.53 (br. s, 6 H, H-2 Man), 3.60 (m, 6 H, H-6
Man), 3.40 (m, 6 H, H-6 Man), 3.50 (m, 12 H, H-3,4 Man), 3.17
(m, H-5 Man) ppm. 13C NMR ([D6]DMSO, 75 MHz): δ = 143.9,
143.7 (C-4 triazole), 124.2, 124.0 (C-5 triazole), 99.7 (C-1 Man),
77.2 (OCH), 74.0, 70.7, 69.9 (C-2,3–5 Man), 66.7 (C-4 Man), 65.0,
64.7, 63.6 (CH2O), 61.0 (C-6 Man), 49.1 (CH2N) ppm. HRMS
(FAB+): calcd. for C72H116N18O42 [M + Na]+ 1927.739; found
1927.788.
Methyl 2,3,4,6-Tetrakis-O-{4-(α-
D-mannopyranosyloxyethyl)-1H-
1,2,3-triazol-1-yl}-α- -glucopyranoside (8Kh): Obtained from 6Kd
D
(504 mg) following the general procedure (Method B). Column
chromatography (MeOH) and lyophilization gave 8Kh as a foamy
solid (505 mg, 90%): [α]2D0 = +61 (c = 1, H O); IR (KBr): ν = 3404,
˜
2
1645, 1134, 1092, 1054 cm–1. 1H NMR ([D6]DMSO, 300 MHz, se-
lected signals): δ = 8.11 (s, 2 H), 8.08 (s, 2 H), 4.84–4.46 (m, 29 H),
4.62 (s, 8 H), 3.94 (m, 4 H), 3.79 (m, 4 H), 3.68–3.17 (m, 30 H),
3.23 (s, 3 H) ppm. 13C NMR ([D6]DMSO, 75 MHz): δ = 144.3,
144.1, 143.9, 143.7, 124.2, 124.1, 99.8, 97.0, 80.9, 79.1, 77.0, 74.1,
70.8, 70.0, 69.6, 68.5, 66.8, 65.5, 65.1, 64.8, 63.8, 63.3, 61.1, 54.4,
49.2 ppm. HRMS (FAB+): calcd. for C51H82N12O30 [M + Na]+
1365.516; found 1365.575.
Enzyme-Linked Lectin Assay (ELLA): ELLA assays were carried
out as described previously.[33] Experiments were carried out using
a Metertech Σ960 instrument. Microtitration plates were coated
with S. cerevisiae mannan with 100 µL/well of a solution of 10 µg/
mL in 10 m phosphate buffer (PBS, pH 7.4) for 2 h at 37 °C.
The wells were then washed twice with 10 m phosphate buffer
containing 1% (v/v) Tween 20 (PBST) and once with PBS. This
4-(α-
kis-O-{4-(α-
D
-Mannopyranosyl)-1H-1,2,3-triazol-1-ylmethyl 2,3,4,6-Tetra-
-mannopyranosyl)-1H-1,2,3-triazol-1-yl}-β- -gluco-
D
D
2452
www.eurjoc.org
© 2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Eur. J. Org. Chem. 2009, 2441–2453

Click Multivalent Homogeneous Neoglycoconjugates
T. Horlacher, J. L. De Paz, P. H. Seeberger, G. G. Hari, K. C.
Mary, A. H. Charles, in Carbohydrate Chemistry, Biology and
Medical Applications (Eds.: H Garg, M. Cowman, C. Hales),
Elsevier, Oxford, 2008, p. 387.
washing procedure was repeated after each incubation period.
Wells were then blocked with 300 µL/well of BSA/PBS (1% w/v)
for 2 h at 37 °C. Each inhibitor (the glycoconjugates or methyl α-
Man) was added in serial dilution (60 µL/well) in PBS (pH 6.8,
containing 0.1 m Ca2+and 0.1 m Mn2+), and the peroxidase-
labeled Con A (60 µL/well of a solution of 50 µg/mL in PBS, pH
6.8, containing 0.1 m Ca2+and 0.1 m Mn2+) was added. The
mixtures of glycoclusters or methyl α-Man and the peroxidase-
labeled lectin (100 µL/well) were added, and the plates were incu-
bated for 2 h at 37 °C. After that, 50 µL/well of a solution of o-
phenylenediamine dihydrochloride (20 mg/50 mL) in citrate-phos-
phate buffer (pH 5.0 with 0.4% H2O2) was added. The plates were
incubated for 30 min at 37 °C. The reactions were stopped by the
addition of aqueous H2SO4 (50 µL/well, 1.25 ), and the ab-
sorbance was measured at 492 nm.
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Received: November 25, 2008
Published Online: April 2, 2009
Eur. J. Org. Chem. 2009, 2441–2453
© 2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.eurjoc.org
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