Synthesis and immunological screening of β-linked mono- and divalent mannosides

Article abstract of DOI:10.1002/ejoc.201200041

Three different β-linked divalent mannosides, along with their corresponding monovalent counterparts, have been designed and chemically synthesized by coupling the corresponding propargyl (propargyl alcohol in the case of the monovalent compounds) and 2-azidoethyl glycosides by using an efficient click chemistry protocol. Crich's β-mannosylation methodology was applied to the construction of the β-mannosidic linkages. All the glycosylation reactions gave moderate-to-good yields with high selectivities. A competitive inhibition enzyme-linked immunosorbent assay (ELISA) was performed to determine the inhibition, by the synthesized mannosides, of specific human IgG binding to low-molecular-weight Candida albicans mannan; moderate inhibition capacity was observed for some of the compounds. Copyright

Full text of DOI:10.1002/ejoc.201200041

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DOI: 10.1002/ejoc.201200041
Synthesis and Immunological Screening of β-Linked Mono- and Divalent
Mannosides
Chinmoy Mukherjee,[a] Kaarina Ranta,[b] Johannes Savolainen,[b] and Reko Leino*[a]
Keywords: Medicinal chemistry / Carbohydrates / Glycosylation / Click chemistry / Biological activity
Three different β-linked divalent mannosides, along with
β-mannosidic linkages. All the glycosylation reactions gave
their corresponding monovalent counterparts, have been de-
moderate-to-good yields with high selectivities. A competi-
signed and chemically synthesized by coupling the corre- tive inhibition enzyme-linked immunosorbent assay (ELISA)
sponding propargyl (propargyl alcohol in the case of the
monovalent compounds) and 2-azidoethyl glycosides by
using an efficient click chemistry protocol. Crich’s β-mannos-
ylation methodology was applied to the construction of the
was performed to determine the inhibition, by the synthe-
sized mannosides, of specific human IgG binding to low-mo-
lecular-weight Candida albicans mannan; moderate inhibi-
tion capacity was observed for some of the compounds.
Introduction
Candida albicans is an opportunistic fungal pathogen
causing systemic candidiasis mostly in immunocom-
promised patients undergoing long-term antibiotic treat-
ment.[1] The chemical structure of the cell wall phos-
phomannan of C. albicans is comprised of β-(1,2)-oligo-
mannosides, which are linked to an α-mannan chain either
through glycosidic bonds or through α-linked phosphodies-
ter bonds (Figure 1).[2,3] The C. albicans β-(1,2)-mannans
are immunogenic and elicit specific antibodies, which have
been shown to be protective against C. albicans in animal
models of systemic and vaginal candidiasis.[4,5] Antibodies
recognize β-mannans on the cell wall of C. albicans de-
pending on the properties and chain lengths of these anti-
genic structures.[6–13]
Figure 1. Partial chemical structure of the cell wall phospho-
mannan of C. albicans with an α-mannan backbone.[2,3]
known reactive groups and structures provides simple tools
for studying these interactions. Overall, synthetic molecules
have been shown to retain the biological properties and an-
tigenicities of their native counterparts.[20,21] In this context
we became interested in studying the biological effects of
the multivalency of different β-linked/β-(1,2)-linked mann-
an constructs of the cell wall of C. albicans, along with
their monovalent counterparts, to develop new tools for
modulating the protective humoral and cellular immune re-
sponses towards Candida.
Divalent model compounds, being the simplest form of
multivalent structures, were selected as a starting point for
this study. To investigate the influence of divalency in bio-
logical systems, comparisons should be made with the cor-
responding monovalent counterparts. For this purpose we
also aimed to synthesize monovalent model compounds.
Various chemical approaches towards divalent sugar deriva-
tives can be envisioned based on, for example, 1) the reac-
tion of 2 equiv. of a sugar unit with a scaffold unit contain-
ing two points of attachment,[22–26] 2) homodimeriza-
Bundle and co-workers[14] have previously shown that
short β-(1,2)-homo-oligosaccharides from disaccharides up
to hexasaccharides inhibit the binding of β-(1,2)-mannan-
specific monoclonal antibodies. The maximum inhibitory
activity was observed for di- and trisaccharides, diminishing
significantly for the corresponding tetra-, penta-, and hexa-
saccharides. It has also been recently shown that a tetrava-
lent β-(1,2)-mannan disaccharide cluster, when linked to a
protein, produces a good antibody response against C. al-
bicans.[15] It is now well understood that molecular recogni-
tion events in biological systems are prone to multivalent
ligand–receptor interactions and that such interactions
often result in higher activities than their monovalent coun-
terparts.[16–19] The chemical synthesis of saccharides with
[a] Laboratory of Organic Chemistry, Åbo Akademi University,
20500 Åbo, Finland
E-mail: reko.leino@abo.fi
[b] Pulmonary Diseases and Clinical Allergology, University of
Turku,
20520 Turku, Finland
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C. Mukherjee, K. Ranta, J. Savolainen, R. Leino
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tion,[26–28] 3) multicomponent reactions,[29] 4) glycosyl- IgG antibodies binding to low-molecular-weight Candida
ation,[25] and 5) heterocoupling between two sugar albicans mannan was performed to determine the inhibition
units.[30–32] The last-mentioned method has successfully by these synthesized mannosides.
been used for the synthesis of divalent carbohydrate model
compounds by using click chemistry[30,33–37] as the synthetic
tool. A beneficial feature of this click approach is its excel-
lent compatibility with a vast range of functional groups,
including alcohols, amines, and carboxylic acids in a
number of solvent systems.[38] As an additional benefit, the
resulting triazole ring is very stable to hydrolytic cleavage
as well as inert towards oxidation and reduction reac-
tions.[39,40] Furthermore, it can also be very effective in con-
trolling the flexibility of the overall system. Triazole-con-
taining compounds also differ from those with other types
of passive linkers and may instead readily associate with
biological targets through hydrogen bonding and dipole in-
teractions.[39] A few literature reports have appeared re-
cently in which the divalent molecules were synthesized by
click coupling of the corresponding propargyl or 3-butynyl
and azidomethyl or 2-azidoethyl glycosides.[41–45] Saccha-
ride ligands, such as mannose (α-linked), glucose, or lactose
units, were used to construct the corresponding 1,2,3-tri-
azole-bridged dimeric compounds. To the best of our
knowledge, similar coupling reactions using β-linked
mannose or mannobiose units have not been explored in
detail.
From another perspective, the chemical synthesis of β-
mannosides has, until recently, remained a challenging task
due to the α-directing anomeric effect and the presence of
an axial substituent at the C-2 position causing steric hin-
drance for the incoming nucleophile. Owing to the pioneer-
ing work of Crich and co-workers and others, several meth-
ods have now become available for the synthesis of β-man-
nosides, either by direct glycosylation[46–51] or by indirect
approaches.[52–58] We successfully used the Crich protocol
in previous work as a direct approach to β-selective manno-
sylation for the construction of several β-linked manno-oli-
gosaccharides.[28,59,60] In this work, we successfully used this
method for the synthesis of carbohydrate building blocks
suitable for click coupling. Thus, herein we report the chem-
ical synthesis of several mono- and divalent β-linked man-
nosides. Furthermore, a competitive inhibition enzyme-
Results and Discussion
Chemistry
Three different monovalent mannosides (1–3), along
with their corresponding divalent counterparts (4–6), were
selected as targets for the synthesis (Figure 2). The sugar
moieties in each of the divalent molecules are connected
through a triazole linker with a specific interconnecting
chain length. Click chemistry was applied to the construc-
tion of the triazole-bridged mono- and divalent mannosides
by reaction of the corresponding propargyl (propargyl
alcohol in the case of the monovalent compounds) and 2-
azidoethyl glycosides. Prior to the coupling reactions, the
glycosides were synthesized by following previously re-
ported literature procedures.
To synthesize the target mono- and divalent mannosides,
three 2-azidoethyl glycosides (10, 12, and 15) and three
propargyl glycosides (20, 21, and 24) were selected as build-
ing blocks (Figure 3). Monosaccharide building block 7,[61]
containing a 4,6-O-benzylidene group and a nonparticipat-
ing 2-O-(p-methoxybenzyl) group (PMB), was used as the
glycosyl donor for introducing the initial β-linkage through
Crich’s methodology for β-mannosylation.
Figure 3. Chemical structures of the building blocks for the click
linked immunosorbent assay (ELISA) of specific human reactions.
Figure 2. Chemical structures of the target mono- and divalent mannosides.
2
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β-Linked Mono- and Divalent Mannosides
In line with this protocol, the glycosyl donor 7 was first pling of each of the azides with propargyl alcohol according
activated in the presence of 1-benzenesulfinylpiperidine to a standard click chemistry protocol by using the copper
(BSP)/2,4,6-tri-tert-butylpyrimidine (TTBP)/Tf2O system at sulfate/sodium ascorbate reagent system. Initially, a test re-
–60 °C to yield a glycosyl triflate; subsequent in situ reac- action was performed in the solvent mixture tBuOH/
tion with 2-azidoethanol[62] at –78 °C furnished the β-mann- H2O,[64] but due to solubility problems the starting materi-
oside 9 in 48% yield (2-azidoethanol is potentially explosive als were first dissolved in CH2Cl2 before adding tBuOH/
and has to be handled at low temperatures). Unlike other H2O (1:1, v/v); the reactions proceeded readily in this
glycosides, there is little difference in the 1-HǞ2-H cou- tBuOH/H2O/CH2Cl2 (1:1:1, v/v/v) solvent system. Follow-
pling constants between the α- and β-mannosides, thus ing this procedure, the coupling products 16–18 were ob-
making difficult the anomeric assignment by 1H NMR tained in yields of 96, 81, and 86%, respectively. Finally,
spectroscopy. The β stereochemistry of 9 was assigned on hydrogenolysis of the cycloaddition products with 10% Pd/
the basis of the upfield 5-H chemical shift at δ = 3.32 ppm C under hydrogen (2.76 bar) furnished the fully deprotected
1
and the JCH value of 154.4 Hz for the anomeric car- monovalent compounds 1–3 in yields of 84, 78, and 82%,
bon.[46,63] Partial degradation of the product was observed respectively (Scheme 2).
during the course of the reaction for an unknown reason,
which reduced the yield. Several precautions were taken and
experiments performed to improve the yield led to an in-
crease in yield of only 2–5%. Deprotection of the p-meth-
oxybenzyl group in glycoside 9 was carried out by treatment
with DDQ to furnish the glycosyl acceptor 10 in good yield.
Again, BSP-mediated β-selective glycosylation of 10 with
glycosyl donor 11[61] afforded the disaccharide 12 in 92%
yield. The formation of disaccharide 12 was confirmed on
the basis of two upfield 5-H chemical shifts at δ = 3.34
1
(5-HA) and 3.35 ppm (5-HB) and the JCH values of 159.4
[1JCH(A)] and 156.5 Hz [1JCH(B)] for the two anomeric car-
Scheme 2. Synthesis of monovalent mannosides 1–3.
bons. Building block 8[61] was readily synthesized from
commercially available d-mannose similarly to compound 7
BSP-mediated β-mannosylation of glycosyl donor 7 with
but subjected to 2-O-acetylation instead of 2-O-p-methoxy- propargyl alcohol afforded the β product 19 in 63% yield.
benzylation. Next, α-selective glycosylation of glycosyl do- The upfield 5-H chemical shift at δ = 3.36 ppm and the
nor 8 with 2-azidoethanol in the presence of N-iodosuccin- 1JCH value of 156.9 Hz for the anomeric carbon indicates
imide/TMSOTf in CH2Cl2 at –40 °C afforded the desired α- the formation of the β product. Deprotection of the p-meth-
glycoside 13 in good yield. By Zemplén deacetylation, 13 oxybenzyl group of glycoside 19 by treatment with DDQ
was converted into the glycosyl acceptor 14 in 90% yield. furnished the glycosyl acceptor 20 in 78% yield. Again,
Finally, BSP-mediated β-mannosylation of 14 with glycosyl BSP-mediated β-selective glycosylation of 20 with glycosyl
donor 11 furnished the desired disaccharide 15 in 88% donor 11 afforded disaccharide 21 in 90% yield. The forma-
yield. An upfield 5-H chemical shift at δ = 3.33 ppm (5-HB) tion of the disaccharide 21 was confirmed on the basis of
1
and JCH values of 170.0 [1JCH(A)] and 154.7 Hz [1JCH(B)
]
two upfield 5-H chemical shifts at δ = 3.38 (5-HA) and
for the anomeric carbons were used to verify the formation 3.35 ppm (5-HB) and the 1JCH values of 158.0 [1JCH(A)] and
of 15 (Scheme 1).
157.7 Hz [1JCH(B)] for the anomeric carbons. Next, α-selec-
tive glycosylation of 8 with propargyl alcohol in the pres-
ence of N-iodosuccinimide/TMSOTf in CH2Cl2 at –40 °C
afforded the desired α-glycoside 22 in 82% yield. Com-
pound 22 was converted into 23 in 91% yield by Zemplén
deacetylation. BSP-mediated β-mannosylation of glycosyl
acceptor 23 with the glycosyl donor 11 furnished the de-
sired disaccharide 24 in 64% yield. The upfield 5-H chemi-
1
cal shift at δ = 3.32 ppm (5-HB) and the JCH values of 169
[1JCH(A)] and 154.6 Hz [1JCH(B)] for the anomeric carbons
suggest the formation of 24 (Scheme 3).
To prepare the divalent mannosides by azido-alkyne cy-
cloaddition, we first tested the reaction by using monosac-
charide building blocks. A robust click coupling reaction
between 2-azidoethyl glycoside 10 and propargyl glycoside
20 provided the expected triazole-fused product 25 in 83%
yield. The coupled product 25 was then subjected to global
Scheme 1. Synthesis of 2-azidoethyl glycosides 10, 12, and 15.
With the three desired 2-azidoethyl glycosides 10, 12, and deprotection under hydrogenolysis in the presence of 10%
15 in hand, we next directed our efforts towards the cou- Pd/C and hydrogen gas (2.76 bar) to furnish the fully depro-
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monovalent counterparts 2 and 3, respectively. However,
relatively lower inhibition was noted for monovalent man-
noside 1 and its divalent counterpart 4.
Scheme 3. Synthesis of the propargyl glycosides 20, 21, and 24.
tected divalent 4 in 86% yield (Scheme 4). After successful
preparation of the divalent mannoside from monosac-
charide building blocks, we then applied the click coupling
method to disaccharides. Accordingly, 2-azidoethyl glycos-
ide 12 and propargyl glycoside 21 were coupled together to
furnish the divalent compound 26 in 86% yield. Subsequent
hydrogenolysis of 26 in the presence of 10% Pd/C under
hydrogen pressure (2.76 bar) afforded the fully deprotected
divalent compound 5 in 97% yield. The divalent molecule 6
was synthesized similarly; the disaccharides 15 and 24 were
coupled to furnish the desired triazole-linked product 27 in
88% yield and subsequent hydrogenolysis of 27 in the
presence of 10% Pd/C under hydrogen (2.76 bar) furnished
the fully deprotected divalent product 6 in 76% yield
(Scheme 4).
Biology
The IgG binding of the synthesized mannosides was
studied through inhibition ELISA experiments with sera
from vaginal candidiasis patients. All study subjects had ele-
vated levels of IgG antibodies against C. albicans mannan.
The binding of these antibodies to Ͻ3 kDa hydrolyzed Can-
dida albicans Cetavlon mannan was inhibited by divalent
mannosides 5 and 6 up to 50% and 70%, respectively (Fig-
ure 4). A similar inhibition pattern was observed by their
Figure 4. Inhibition of IgG binding to low-molecular-weight
(Ͻ3 kDa) hydrolyzed Cetavlon C. albicans mannan by synthetic
mono- and divalent mannosides in ELISA. C. albicans mannan
(autoinhibition) is included as a control. The degree of inhibition
in various concentrations is shown in the y axis and is defined by
the formula 100 – {[1 – (absorbance of sera treated with various
concentrations of saccharide inhibitors– absorbance of PBS)/ab-
sorbance of sera]}ϫ100. Each line represents the results from one
study subject.
Scheme 4. Synthesis of divalent mannosides 4–6.
4
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β-Linked Mono- and Divalent Mannosides
niques used.[65] HRMS were recorded by using either a Bruker
Micro Q-TOF with ESI (electrospray ionization) spectrometer op-
erated in positive mode or a Fison ZabSpecOaTOF spectrometer
with EI (electron impact) ionization operated in positive mode. Op-
tical rotations were measured with a Perkin–Elmer 241 polarimeter
equipped with a Na lamp (589 nm). Reactions were monitored by
TLC on aluminium sheets precoated with silica gel 60 F254
(Merck). The compounds were detected under UV light and by
charring with H2SO4/MeOH (1:10, v/v). Column chromatography
was performed on silica gel 60 (0.040–0.060 mm, Merck) by using
Previous studies have shown that β-(1,2)-linked manno-
biose, which consists of two β-anomeric linkages, is consid-
ered to be the minimum requirement for inhibiting the
binding of Candida-specific antibodies.[14,15] In this study,
divalent mannoside 5, with two β-linkages, showed higher
inhibitory activity than its corresponding α-linked (at the
reducing terminus) counterpart 6. It is also evident that di-
valency appears not to be essential for inhibition as no dif-
ferences in the inhibition of IgG binding with divalent com-
pounds were observed in comparison with monovalent different solvent systems based on the polar character of different
compounds.
compounds. In contrast, the β-linked monosaccharide com-
pound 1 and its divalent analogue 4 showed low inhibitory
activity. Thus, the biological activity declines sharply in the
following order: disaccharide (with two β linkages) Ͼ disac-
charide (with one α and one β linkage) Ͼ monosaccharide
(β-linkage). Our results are in accordance with earlier find-
ings and confirm the potential antigenic properties of β-
(1,2)-linked Candida albicans derived mannans.
General Procedure A. Typical Procedure for the Synthesis of β-
Mannosylation Reaction: Molecular sieves (4 Å, 500 mg) were
added to a solution of compound 11 (164 mg, 0.30 mmol) in
CH2Cl2 (5 mL) and the mixture was stirred for 30 min at room
temperature under argon. BSP (76 mg, 0.36 mmol) and TTBP
(113 mg, 0.46 mmol) were added at –60 °C followed by the addition
of Tf2O (111 mg, 66 μL, 0.39 mmol). The resulting mixture was
stirred under the same conditions for 30 min. Acceptor 10 (100 mg,
0.23 mmol) in CH2Cl2 (2 mL) was then added to the reaction mix-
ture at –78 °C and stirred for 2 h at this temperature before quench-
ing with Et3N (200 μL). The reaction mixture was then filtered
through a pad of Celite followed by washing with CH2Cl2 (30 mL).
The eluted organic solvent was washed with water (15 mL), satd.
aq. NaHCO3 (15 mL), and brine (15 mL), dried (Na2SO4), concen-
trated, and purified over SiO2 to give 12 (185 mg, 92%). Com-
pounds 9, 15, 19, 21, and 24 were then synthesized following an
analogous procedure.
Conclusions
Three structurally unique divalent mannosides have been
designed on the basis of immunopotent saccharides from
Candida albicans mannan and synthesized for the first time
by using click chemistry to study the effects of multivalency,
length of saccharide chain, and linkage type on immune
responses. Crich’s β-mannosylation methodology was suc-
cessfully used for the construction of the β-mannosidic link-
ages. Divalent compounds 5 and 6 were found to possess
similar properties important in the antigenicity of C. al-
bicans, as these compounds inhibited specific IgG antibody
General Procedure B. Typical Experimental Procedure for the Click
Reaction: Copper sulfate (8 mg, 0.05 mmol) and sodium ascorbate
(20 mg, 0.10 mmol) were added to a solution of compound 10
(74 mg, 0.17 mmol) and propargyl alcohol (20 μL, 19 mg,
0.34 mmol) in tBuOH/H2O/CH2Cl2 (1:1:1, v/v/v, 2.4 mL). The re-
binding to Ͻ3 kDa hydrolyzed mannan up to 50% and sulting reaction mixture was stirred at room temperature for 16 h.
Satd. NH4Cl/water (1:1, v/v, 4 mL) was poured into the solution
and the mixture was extracted with CH2Cl2 (3ϫ10 mL). The com-
bined organic layers were washed with water (10 mL) and brine
(10 mL), and dried (Na2SO4), filtered, concentrated, and purified
over SiO2 to afford the title compound 16 (80 mg, 96%). Com-
pounds 17, 18, 25, 26, and 27 were then synthesized following an
analogous procedure.
70%. The findings of this study confirm that the two syn-
thesized divalent mannosides (compounds 5 and 6) share
some similarities with the immunopotent and antigenic epi-
topes of β-(1,2)-linked Candida albicans mannan. However,
divalency appears not to strengthen the inhibition capacity
of the mannosides. Further synthetic and biological studies
on these and similar compounds are ongoing in our
laboratories and will be reported in more detail in forth-
coming papers.
General Procedure C. Typical Experimental Procedure for the Hy-
drogenolysis Reaction: Pd/C (10%, 120 mg) was added to a solution
of compound 16 (60 mg, 0.12 mmol) in methanol/ethyl acetate (5:1,
v/v, 3 mL) and the mixture was stirred under H2 (2.76 bar) for 16 h
at room temperature. The reaction mixture was filtered through a
pad of Celite and washed with methanol (3ϫ10 mL). The com-
bined filtrate was concentrated under reduced pressure to afford
pure 1 (32 mg, 84%). Compounds 2–6 were then synthesized fol-
lowing an analogous procedure.
Experimental Section
General Methods: All reaction solvents were distilled and dried un-
der argon before use and all reactions were carried out under argon
unless noted otherwise. Molecular sieves (4 Å) were activated under
vacuum at 300 °C for 1 h prior to use in the reactions. All NMR
spectra were recorded with a Bruker Avance spectrometer operating
2-Azidoethyl 3-O-Benzyl-4,6-O-benzylidene-2-O-(p-methoxybenzyl)-
β-D-mannopyranoside (9): The glycosidation reaction was carried
1
at 600.13 MHz for H and 150.90 MHz for 13C. All products were
out starting from glycosyl donor 7 (700 mg, 1.23 mmol) and 2-az-
idoethanol (129 mg, 1.48 mmol) following the same reaction pro-
cedure as outlined in General Procedure A to afford compound 9
fully characterized by using 1D 1H and 13C NMR techniques in
combination with 2D COSY, HSQC (both coupled and decoupled),
and HMBC NMR techniques at 25 °C. Chemical shifts are re-
ported downfield from TMS with residual chloroform or methanol
(320 mg, 48%). [α]2D5
= –74.3 (c =
1, CHCl3). 1H NMR
(600.13 MHz, CDCl3): δ = 7.51–7.49 (m, 2 H, Ar-H), 7.40–7.25 (m,
10 H, Ar-H), 6.86–6.84 (m, 2 H, Ar-H), 5.62 (s, 1 H, PhCH), 4.90,
4.79 (2 d, J = –11.9 Hz each, 2ϫ1 H, PhCH2), 4.65, 4.55 (2 d, J
= –12.5 Hz, 2ϫ1 H, PhCH2), 4.53 (d, J1,2 = 1.0 Hz, 1 H, 1-H),
1
as the internal reference. Computational analysis of the H NMR
spectra of all the mono- and disaccharide building blocks was
achieved by using PERCH NMR software with the starting values
and spectral parameters obtained from the various NMR tech-
4.30 (dd, J6a,6b = –10.3, J5,6a = 4.8 Hz, 1 H, 6-Ha), 4.19 (dd, J3,4
=
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9.9, J4,5 = 9.2 Hz, 1 H, 4-H), 4.11 (ddd, JOCH2a,OCH2b = –10.6,
JOCH2a,CH2bN3 = 4.5, JOCH2a,CH2aN3 = 3.5 Hz, 1 H, OCH2a), 3.99
(dd, J2,3 = 3.1 Hz, 1 H, 2-H), 3.93 (dd, J5,6b = 10.1 Hz, 1 H, 6-Hb),
3.80 (s, 3 H, OCH3), 3.65 (ddd, JOCH2b,CH2aN3 = 8.7, JOCH2b,CH2bN3
= 3.2 Hz, 1 H, OCH2b), 3.58 (dd, 1 H, 3-H), 3.56 (ddd,
tion of compound 8 (1.5 g, 3.05 mmol) and 2-azidoethanol
(320 mg, 3.65 mmol) in CH2Cl2 (20 mL). The reaction mixture was
then stirred at room temperature for 30 min under argon. NIS
(825 mg, 3.66 mmol) was added and the reaction mixture was co-
oled to –40 °C. TMSOTf (81 mg, 66 μL, 0.37 mmol) was then
JCH2aN3,CH2bN3 = –13.4 Hz, 1 H, CH2aN3), 3.34 (ddd, 1 H, added and the reaction mixture was stirred for 1 h at the same
CH2bN3), 3.33 (ddd, 1 H, 5-H) ppm. 13C NMR (150.90 MHz, temperature and quenched with Et3N (200 μL), filtered through a
CDCl3): δ = 159.2–113.5 (Ar-C), 102.3 (C-1), 101.4 (PhCH), 78.5
pad of Celite, and washed with CH2Cl2 (60 mL). The eluted or-
(C-4), 77.7 (C-3), 75.3 (C-2), 74.6 (PhCH2), 72.3 (PhCH2), 68.7 ganic solvent was washed with water (30 mL), a satd. aq. NaHCO3
(OCH2), 68.5 (C-6), 67.7 (C-5), 55.2 (OCH3), 50.8 (CH2N3) ppm. solution (30 mL), a 10% aq. Na2S2O3 solution (30 mL), and brine
HRMS (ESI): calcd. for C30H33N3O7Na [M + Na]+ 570.2216;
found 570.2218.
(2ϫ30 mL). The organic layer was then dried (Na2SO4), concen-
trated, and purified over SiO2 to afford pure 13 (1.23 g, 86%) as a
1
colorless oil. [α]2D5 = +6.4 (c = 1, CHCl3). H NMR (600.13 MHz,
2-Azidoethyl 3-O-Benzyl-4,6-O-benzylidene-β-D-mannopyranoside
CDCl3): δ = 7.51–7.24 (m, 10 H, Ar-H), 5.63 (s, 1 H, PhCH), 5.41
(dd, J2,3 = 3.5, J1,2 = 1.6 Hz, 1 H, 2-H), 4.84 (d, 1 H, 1-H), 4.69,
4.66 (2 d, J = –12.1 Hz each, PhCH2), 4.27 (dd, J6a,6b = –10.4, J5,6a
= 4.9 Hz, 1 H, 6-Ha), 4.07 (dd, J3,4 = 10.0, J4,5 = 9.4 Hz, 1 H, 4-
H), 4.04 (dd, 1 H, 3-H), 3.88 (ddd, J5,6b = 10.3 Hz, 1 H, 5-H), 3.86
(dt, JOCH2a,OCH2b = –10.6, JOCH2a,CH2N3 = 5.0 Hz, 1 H, OCH2a),
3.85 (dd, 1 H, 6-Hb), 3.60 (dt, JOCH2b,CH2N3 = 5.0 Hz, 1 H,
OCH2b), 3.39 (dd, 2 H, CH2N3), 2.16 (s, 3 H, COCH3) ppm. 13C
NMR (150.90 MHz, CDCl3): δ = 170.1 (CO), 137.8–126.0 (Ar-C),
101.6 (PhCH), 98.9 (C-1), 78.1 (C-4), 73.6 (C-3), 72.2 (PhCH2),
69.5 (C-2), 68.6 (C-6), 66.9 (OCH2), 64.2 (C-5), 50.3 (CH2N3) ppm.
HRMS (ESI): calcd. for C24H27N3O7Na [M + Na]+ 492.1747;
found 492.1730.
(10): DDQ (160 mg, 0.70 mmol) was added to a solution of com-
pound 9 (320 mg, 0.58 mmol) in CH2Cl2/H2O (1:1, v/v, 20 mL).
The reaction mixture was stirred at room temperature for 2 h, di-
luted with CH2Cl2 (20 mL), and washed with water (2ϫ20 mL)
and satd. aq. NaHCO3 (2ϫ20 mL). The organic layer was then
dried (Na2SO4), filtered, concentrated, and purified over SiO2 to
afford pure 10 (220 mg, 88%) as a colorless oil. [α]2D5 = –40.1 (c =
1, CHCl3). H NMR (600.13 MHz, CDCl3): δ = 7.51–7.25 (m, 10
H, Ar-H), 5.61 (s, 1 H, PhCH), 4.86, 4.78 (2 d, J = –12.2 Hz each,
2ϫ1 H, PhCH2), 4.58 (d, J1,2 = 1.1 Hz, 1 H, 1-H), 4.33 (dd, J6a,6b
1
= –10.5, J5,6a = 4.9 Hz, 1 H, 6-Ha), 4.18 (ddd, J2,3 = 3.3, J2,2-OH
1.6 Hz, 1 H, 2-H), 4.16 (dd, J3,4 = 9.5, J4,5 = 9.4 Hz, 1 H, 4-H), 4.09
(ddd, JOCH2a,OCH2b = –10.8, JOCH2a,CH2bN3 = 5.0, JOCH2a,CH2aN3
=
=
2-Azidoethyl 3-O-Benzyl-4,6-O-benzylidene-α-D-mannopyranoside
3.8 Hz, 1 H, OCH2a), 3.89 (dd, J5,6b = 10.1 Hz, 1 H, 6-Hb), 3.74
(ddd, JOCH2b,CH2aN3 = 8.2, JOCH2b,CH2bN3 = 3.6 Hz, 1 H, OCH2b),
3.66 (dd, 1 H, 3-H), 3.54 (ddd, JCH2aN3,CH2bN3 = –13.4 Hz, 1 H,
CH2aN3), 3.38 (ddd, 1 H, CH2bN3), 3.36 (ddd, 1 H, 5-H), 2.56 (d,
1 H, 2-OH) ppm. 13C NMR (150.90 MHz, CDCl3): δ = 137.8–
126.0 (Ar-C), 101.5 (PhCH), 100.6 (C-1), 78.2 (C-4), 76.5 (C-3),
72.5 (PhCH2), 69.7 (C-2), 68.6 (OCH2), 68.5 (C-6), 67.0 (C-5), 50.6
(CH2N3) ppm. HRMS (ESI): calcd. for C22H25N3O6Na [M +
Na]+ 450.1641; found 450.1637.
(14): Sodium methoxide (1 m in MeOH, pH ca. 12) was added to
a solution of compound 13 (460 mg, 0.98 mmol) in MeOH (10 mL)
and the resulting mixture was stirred at room temperature for 2 h.
Dowex 50WX8-100 (H+) was then added to neutralize the solution.
The reaction mixture was filtered and washed with methanol
(3ϫ20 mL). The combined organic layers were evaporated to dry-
ness under reduced pressure and purified over SiO2 to afford pure
14 (375 mg, 90%) as a white foam. [α]2D5 = +27.5 (c = 1, CHCl3).
1H NMR (600.13 MHz, CDCl3): δ = 7.50–7.25 (m, 10 H, Ar-H),
5.62 (s, 1 H, PhCH), 4.92 (d, J1,2 = 1.5 Hz, 1 H, 1-H), 4.86, 4.72
(2 d, J = –11.8 Hz each, 2ϫ1 H, PhCH2), 4.27 (dd, J6a,6b = –10.7,
J5,6a = 5.3 Hz, 1 H, 6-Ha), 4.11 (dd, J3,4 = 9.6, J4,5 = 9.2 Hz, 1 H,
4-H), 4.10 (ddd, J2,3 = 3.4, J2,2-OH = 1.4 Hz, 1 H, 2-H), 3.95 (dd,
1 H, 3-H), 3.90 (ddd, JOCH2a,OCH2b = –10.7, JOCH2a,CH2aN3 = 6.5,
JOCH2a,CH2bN3 = 3.3 Hz, 1 H, OCH2a), 3.86 (dd, J5,6b = 10.2 Hz, 1
H, 6-Hb), 3.86 (ddd, 1 H, 5-H), 3.63 (ddd, JOCH2b,CH2bN3 = 6.8,
JOCH2b,CH2aN3 = 3.3 Hz, 1 H, OCH2b), 3.42 (ddd, JCH2aN3,CH2bN3
= –13.3 Hz, 1 H, CH2aN3), 3.38 (ddd, 1 H, CH2bN3), 2.70 (d, 1 H,
2-OH) ppm. 13C NMR (150.90 MHz, CDCl3): δ = 137.9–126.0 (Ar-
C), 101.6 (PhCH), 100.1 (C-1), 78.6 (C-4), 75.4 (C-3), 73.1
(PhCH2), 69.8 (C-2), 68.8 (C-6), 66.7 (OCH2), 63.6 (C-5), 50.4
(CH2 N3 ) ppm. HRMS (ESI): calcd. for C2 2 H2 5 N3 O6 Na
[M + Na]+ 450.1641; found 450.1646.
2-Azidoethyl (2,3-Di-O-benzyl-4,6-O-benzylidene-β-D-mannopyran-
osyl)-(1Ǟ2)-3-O-benzyl-4,6-O-benzylidene-β- -mannopyranoside
D
(12): [α]2D5 = –102.9 (c = 1, CHCl3). 1H NMR (600.13 MHz,
CDCl3): δ = 7.58–7.24 (m, 25 H, Ar-H), 5.61 (s, 1 H, PhCH), 5.43
(s, 1 H, PhCH), 5.08, 5.01 (2 d, J = –12.3 Hz each, 2 ϫ 1 H,
PhCH2), 4.79 (d, J1B,2B = 0.9 Hz, 1 H, 1-HB), 4.79, 4.77 (2 d, J =
–12.6 Hz each, 2ϫ1 H, PhCH2), 4.66, 4.58 (2 d, J = –12.6 Hz each,
2ϫ1 H, PhCH2), 4.52 (s, 1 H, 1-HA), 4.31 (dd, J6aB,6bB = –10.4,
J5B,6aB = 4.8 Hz, 1 H, 6-HaB), 4.28 (dd, J6aA,6bA = –10.4, J5A,6aA
4.8 Hz, 1 H, 6-HaA), 4.27 (dd, J2A,3A = 3.1 Hz, 1 H, 2-HA), 4.23
(dd, J3B,4B = 10.0, J4B,5B = 9.2 Hz, 1 H, 4-HB), 4.14 (dd, J2B,3B
3.2 Hz, 1 H, 2-HB), 4.07 (ddd, JOCH2a,OCH2b = –10.4, JOCH2a,CH2bN3
= 4.9, JOCH2a,CH2aN3 = 3.2 Hz, 1 H, OCH2a), 4.06 (dd, J3A,4A
=
=
=
9.9, J4A,5A = 9.3 Hz, 1 H, 4-HA), 3.90 (dd, J5B,6bB = 10.2 Hz, 1
H, 6-HbB), 3.75 (dd, J5A,6bA = 10.0 Hz, 1 H, 6-HbA), 3.66 (ddd,
JOCH2b,CH2aN3 = 8.5, JOCH2b,CH2bN3 = 2.8 Hz, 1 H, OCH2b), 3.62
(dd, 1 H, 3-HA), 3.59 (dd, 1 H, 3-HB), 3.35 (ddd, 1 H, 5-HB), 3.34
(ddd, 1 H, 5-HA), 3.27 (ddd, JCH2aN3,CH2bN3 = –13.4 Hz, 1 H,
CH2aN3), 3.19 (ddd, 1 H, CH2bN3) ppm. 13C NMR (150.90 MHz,
CDCl3): δ = 139.0–126.0 (Ar-C), 103.7 (C-1B), 101.5 (C-1A), 101.5
(PhCH), 101.3 (PhCH), 78.4 (C-4B), 78.1 (C-4A), 77.3 (C-3B), 76.8
(C-2A), 75.9 (C-3A), 75.8 (C-2B), 74.5 (PhCH2), 71.1 (PhCH2), 68.7
(OCH2), 68.7 (C-6B), 68.6 (C-6A), 67.6 (C-5B), 67.5 (C-5A), 50.8
(CH2N3) ppm. HRMS (ESI): calcd. for C49H55N4O11 [M + NH4]+
875.3867; found 875.3841.
2-Azidoethyl (2,3-Di-O-benzyl-4,6-O-benzylidene-β-D-mannopyran-
osyl)-(1Ǟ2)-3-O-benzyl-4,6-O-benzylidene-α- -mannopyranoside
D
(15): Employing the same method as outlined in General Procedure
A, the glycosylation reaction between glycosyl donor 11 (164 mg,
0.30 mmol) and glycosyl acceptor 14 (100 mg, 0.23 mmol) gave
compound 15 (177 mg, 88%). [α]2D5 = –50.3 (c = 1, CHCl3). 1H
NMR (600.13 MHz, CDCl3): δ = 7.53–7.25 (m, 25 H, Ar-H), 5.60
(s, 1 H, PhCH), 5.50 (s, 1 H, PhCH), 5.04, 4.98 (2 d, J = –12.2 Hz
each, 2ϫ1 H, PhCH2), 4.86 (d, J1A,2A = 1.7 Hz, 1 H, 1-HA), 4.76,
4.73 (2 d, J = –12.2 Hz each, 2ϫ1 H, PhCH2), 4.71, 4.62 (2 d, J
= –12.5 Hz each, 2ϫ1 H, PhCH2), 4.63 (s, 1 H, 1-HB), 4.27 (dd,
2-Azidoethyl 2-O-Acetyl-3-O-benzyl-4,6-O-benzylidene-α-
D-manno-
J2A,3A = 3.3 Hz, 1 H, 2-HA), 4.27 (dd, J6aB,6bB = –10.5, J5B,6aB
=
pyranoside (13): Molecular sieves (4 Å, 3 g) were added to a solu-
4.8 Hz, 1 H, 6-HaB), 4.26 (dd, J3B,4B = 9.8, J4B,5B = 9.3 Hz, 1 H,
6
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© 0000 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Eur. J. Org. Chem. 0000, 0–0

Job/Unit: O20041
/KAP1
Date: 10-04-12 15:13:49
Pages: 13
β-Linked Mono- and Divalent Mannosides
4-HB), 4.25 (dd, J6aA,6bA = –10.3, J5A,6aA = 4.7 Hz, 1 H, 6-HaA),
4.12 (dd, J3A,4A = 10.0, J4A,5A = 9.5 Hz, 1 H, 4-HA), 3.99 (dd,
J2B,3B = 3.2 Hz, 1 H, 2-HB), 3.98 (dd, 1 H, 3-HA), 3.88 (ddd,
≈ J4B,5B ≈ 9.7 Hz, 1 H, 4-HB), 4.20 (d, J2A,3A = 3.2 Hz, 1 H, 2-HA),
4.02–3.97 (m, 3 H, 4-HA, 2-HB, CH2bCH2 triazole), 3.88 (dd,
J5B,6bB ≈ J6aB,6bB ≈ 10.3 Hz, 1 H, 6-HbB), 3.75 (dd, J5A,6bA
J6aA,6bA ≈ 10.2 Hz, 1 H, 6-HbA), 3.65 (dd, J2B,3B = 3.2 Hz, 1 H, 3-
3.3 Hz, 1 H, OCH2a), 3.87 (dd, J5B,6bB = 10.0 Hz, 1 H, 6-HbB), 3.82 HB), 3.58 (dd, J3A,4A = 9.9 Hz, 1 H, 3-HA), 3.33 (ddd, J4A,5A = 9.8,
≈
JOCH2a,OCH2b = –10.7, JOCH2a,CH2bN3 = 5.7, JOCH2a,CH2aN3
=
(ddd, J5A,6bA = 10.3 Hz, 1 H, 5-HA), 3.77 (dd, 1 H, 6-HbA), 3.60
(dd, 1 H, 3-HB), 3.59 (ddd, JOCH2b,CH2aN3 = 7.5, JOCH2b,CH2bN3
3.2 Hz, 1 H, OCH2b), 3.43 (ddd, JCH2aN3,CH2bN3 = –13.3 Hz, 1 H,
J5A,6aA = 4.8 Hz, 1 H, 5-HA), 3.25 (ddd, J5B,6aB = 4.9 Hz, 1 H, 5-
HB) ppm. 13C NMR (150.90 MHz, CDCl3): δ = 148.0 (C-4, tri-
azole), 139.0–126.0 (30 C, Ar-C), 121.9 (C-5, triazole), 103.2 (C-
=
CH2aN3), 3.35 (ddd, 1 H, CH2bN3), 3.33 (ddd, 1 H, 5-HB) ppm. 1B), 101.6 (C-1A), 101.5 (PhCH), 101.4 (PhCH), 78.7 (C-4B), 78.0
13C NMR (150.90 MHz, CDCl3): δ = 138.8–126.0 (Ar-C), 101.6
(C-4A), 77.7 (C-3B), 76.0 (C-2B), 75.7 (C-3A), 75.6 (C-2A), 74.5
(PhCH), 101.4 (PhCH), 100.9 (C-1B), 98.7 (C-1A), 78.5 (C-2A), 78.4 (PhCH2), 72.3 (PhCH2), 71.0 (PhCH2), 68.7 (C-6A), 68.6 (C-6B),
(C-4A), 77.6 (C-3B), 75.9 (C-3A), 75.0 (C-4B), 74.5 (PhCH2), 74.0 67.7 (CH2CH2 triazole), 67.6 (C-5A), 67.4 (C-5B), 56.6 (CH2OH),
(C-2B), 72.3 (PhCH2), 71.5 (PhCH2), 68.8 (C-6A), 68.5 (C-6B), 67.8 50.1 (CH2 CH2 triazole) ppm. HRMS (ESI): calcd. for
(C-5B), 66.6 (OCH2), 64.5 (C-5A), 50.4 (CH2N3) ppm. HRMS
(ESI): calcd. for C49H51N3O11Na [M + Na]+ 880.3421; found
880.3400.
C52H55N3O12Na [M + Na]+ 936.3683; found 936.3649.
1-[(β- -Mannopyranosyl)-(1Ǟ2)-(β- -mannopyranosyloxyethyl)]-
D
D
4-hydroxymethyl-1,2,3-triazole (2): Employing the same method as
outlined in General Procedure C, hydrogenolysis of the coupling
product 17 (30 mg, 0.03 mmol) gave compound 2 (12 mg, 78%).
1-(3-O-Benzyl-4,6-O-benzylidene-β-D-mannopyranosyloxyethyl)-
4-hydroxymethyl-1,2,3-triazole (16): [α]2D5 = –16.8 (c = 1, CHCl3).
1H NMR (600.13 MHz, CDCl3): δ = 7.74 (s, 1 H, triazole-H), 7.49–
7.46 (m, 2 H, Ar-H), 7.39–7.24 (m, 8 H, Ar-H), 5.58 (s, 1 H,
PhCH), 4.80, 4.73 (2 d, J = –12.2 Hz each, 2ϫ1 H, PhCH2), 4.71
(s, 2 H, CH2abOH), 4.58–4.49 (m, 2 H, CH2CH2ab triazole), 4.42
(s, 1 H, 1-H), 4.27 (dd, J6a,6b = –10.4, J5,6a = 4.9 Hz, 1 H, 1-H),
4.21 (ddd, J = –10.9, 4.8, 3.7 Hz, 1 H, CH2aCH2 triazole), 4.10 (dd,
J3,4 ≈ J4,5 ≈ 9.5 Hz, 1 H, 4-H), 4.07 (d, J2,3 = 3.1 Hz, 1 H, 2-H),
3.90 (ddd, J = 8.0, 3.6 Hz, 1 H, CH2bCH2 triazole), 3.84 (dd, J5,6b
= 10.3 Hz, 1 H, 6-Hb), 3.60 (dd, 1 H, 3-H), 3.30 (ddd, 1 H, 5-
H) ppm. 13C NMR (150.90 MHz, CDCl3): δ = 147.7 (C-4, triazole),
137.7–125.9 (12 C, Ar-C), 123.4 (C-5, triazole), 101.4 (PhCH),
100.7 (C-1), 78.2 (C-4), 76.6 (C-3), 72.4 (PhCH2), 69.3 (C-2), 68.3
(C-6), 67.9 (CH2CH2 triazole), 66.9 (C-5), 56.0 (CH2OH), 50.1
(CH2CH2 triazole) ppm. HRMS (ESI): calcd. for C25H29N3O7Na
[M + Na]+ 506.1903; found 506.1886.
1
[α]2D5 = –40.6 (c = 1, H2O). H NMR (600.13 MHz, CD3OD): δ =
7.99 (s, 1 H, triazole-H), 4.72 (br. s, 2 H, CH2abOH), 4.67–4.62 (m,
3 H, 1-HA, CH2CH2ab triazole), 4.58 (d, J1B,2B = 0.5 Hz, 1 H, 1-
HB), 4.31 (ddd, J = –11.0, 5.6, 3.8 Hz, 1 H, CH2aCH2 triazole),
4.05 (d, J2A,3A = 3.2 Hz, 1 H, 2-HA), 4.00 (ddd, J = 6.8, 3.7 Hz, 1
H, CH2bCH2 triazole), 3.89 (dd, J6aA,6bA = –11.8, J5A,6aA = 2.2 Hz,
1 H, 6-HaA), 3.86 (dd, J6aB,6bB = –12.0, J5B,6aB = 2.2 Hz, 1 H, 6-
HaB), 3.78 (d, J2B,3B = 2.5 Hz, 1 H, 2-HB), 3.70 (dd, J5A,6bA
=
6.1 Hz, 1 H, 6-HbA), 3.66 (dd, J5B,6bB = 6.6 Hz, 1 H, 6-HbB), 3.50
(dd, J3A,4A ≈ J4A,5A ≈ 9.6 Hz, 1 H, 4-HA), 3.49 (dd, J3B,4B ≈ J4B,5B
≈ 9.6 Hz, 1 H, 4-HB), 3.44 (2 dd, 2 H, 3-HA, 3-HB), 3.22 (ddd, 1
H, 5-HA), 3.15 (ddd, 1 H, 5-HB) ppm. 13C NMR (150.90 MHz,
CD3OD): δ = 149.2 (C-4, triazole), 124.8 (C-5, triazole), 102.3 (C-
1A), 101.9 (C-1B), 78.7 (C-5A), 78.6 (C-5B), 78.5 (C-2A), 75.1 (C-3B),
74.5 (C-3A), 72.2 (C-2B), 69.0 (C-4A), 68.9 (C-4B), 68.7 (CH2CH2
triazole), 63.1 (C-6B), 62.8 (C-6A), 56.6 (CH2OH), 51.9 (CH2CH2
triazole) ppm. HRMS (ESI): calcd. for C17 H2 9N3O12 Na
[M + Na]+ 490.1649; found 490.1653.
1-(β-D-Mannopyranosyloxyethyl)-4-hydroxymethyl-1,2,3-triazole
1
(1): [α]2D5 = –22.8 (c = 1, H2O). H NMR (600.13 MHz, CD3OD):
δ = 8.06 (s, 1 H, triazole-H), 4.68 (br. s, 2 H, CH2abOH), 4.66–4.64
(m, 2 H, CH2CH2ab triazole), 4.52 (d, J1,2 = 1.0 Hz, 1 H, 1-H),
4.27 (ddd, J = –11.3, 5.3, 4.1 Hz, 1 H, CH2aCH2 triazole), 4.00
1-[(2,3-Di-O-benzyl-4,6-O-benzylidene-β-
D-mannopyranosyl)-
(1Ǟ2)-(3-O-benzyl-4,6-O-benzylidene-α- -mannopyranosyloxy-
D
ethyl)]-4-hydroxymethyl-1,2,3-triazole (18): The click coupling reac-
tion was carried out starting from 2-azidoethyl glycoside 15 (72 mg,
0.08 mmol) and propargyl alcohol (10 μL, 9.4 mg, 0.17 mmol) fol-
lowing the same method as outlined in General Procedure B to
afford the title compound 18 (66 mg, 86%). [α]2D5 = –40.1 (c = 1,
(ddd, J = 6.5, 4.3 Hz, 1 H, CH2bCH2 triazole), 3.88 (dd, J6a,6b
–11.8, J5,6a = 2.3 Hz, 1 H, 6-Ha), 3.83 (dd, J2,3 = 3.2 Hz, 1 H, 2-
H), 3.70 (dd, J5,6b = 6.1 Hz, 1 H, 6-Hb), 3.54 (dd, J3,4 ≈ J4,5
=
≈
9.5 Hz, 1 H, 4-H), 3.43 (dd, 1 H, 3-H), 3.22 (ddd, 1 H, 5-H) ppm.
13C NMR (150.90 MHz, CD3OD): δ = 148.9 (C-4, triazole), 125.3
(C-5, triazole), 101.9 (C-1), 78.4 (C-5), 75.1 (C-3), 72.3 (C-2), 68.9
(CH2CH2 triazole), 68.5 (C-4), 62.8 (C-6), 56.5 (CH2OH), 51.6
(CH2CH2 triazole) ppm. HRMS (ESI): calcd. for C11H19N3O7Na
[M + Na]+ 328.1121; found 328.1119.
1
CHCl3). H NMR (600.13 MHz, CDCl3): δ = 7.50–7.24 (m, 26 H,
Ar-H, triazole-H), 5.59 (s, 1 H, PhCH), 5.44 (s, 1 H, PhCH), 4.99,
4.93 (2 d, J = –12.2 Hz each, 2ϫ1 H, PhCH2), 4.75 (br. s, 2 H,
PhCH2), 4.69 (d, J = –12.5 Hz, 1 H, PhCH2), 4.68 (br. s, 1 H, 1-
HA), 4.65 (br. s, 2 H, CH2abOH), 4.60 (d, J = –12.5 Hz, 1 H,
PhCH2), 4.56 (br. s, 1 H, 1-HB), 4.50–4.46 (m, 2 H, CH2CH2ab
1-[(2,3-Di-O-benzyl-4,6-O-benzylidene-β-
(1Ǟ2)-(3-O-benzyl-4,6-O-benzylidene-β-
D-mannopyranosyl)-
D
-mannopyranosyloxy- triazole), 4.27 (dd, J6aB,6bB = –10.3, J5B,6aB = 4.8 Hz, 1 H, 6-HaB),
ethyl)]-4-hydroxymethyl-1,2,3-triazole (17): Employing the same
method as outlined in General Procedure B, the click coupling re-
action between 2-azidoethyl glycoside 12 (37 mg, 0.04 mmol) and
propargyl alcohol (5 μL, 5 mg, 0.09 mmol) gave compound 17
(32 mg, 81%). [α]2D5 = –67.8 (c = 1, CHCl3). 1H NMR (600.13 MHz,
CDCl3): δ = 7.57–7.21 (m, 26 H, Ar-H, triazole-H), 5.60 (s, 1 H,
PhCH), 5.43 (s, 1 H, PhCH), 5.05, 4.97 (2 d, J = –12.4 Hz each,
4.24 (dd, J3B,4B ≈ J4B,5B ≈ 9.6 Hz, 1 H, 4-HB), 4.11 (dd, J1A,2A =
1.6, J2A,3A = 3.4 Hz, 1 H, 2-HA), 4.09 (dd, J6aA,6bA = –10.2, J5A,6aA
= 4.7 Hz, 1 H, 6-HaA), 4.06–4.01 (m, 2 H, 4-HA, CH2aCH2 tri-
azole), 3.94 (d, J2B,3B = 3.1 Hz, 1 H, 2-HB), 3.87 (dd, J5B,6bB
=
10.2 Hz, 1 H, 6-HbB), 3.82–3.76 (m, 2 H, 3-HA, CH2bCH2 triazole),
3.66 (dd, J5A,6bA = 10.3 Hz, 1 H, 6-HbA), 3.59 (dd, 1 H, 3-HB), 3.33
(ddd, 1 H, 5-HB), 3.07 (ddd, 1 H, 5-HA) ppm. 13 C NMR
2 ϫ 1 H, PhCH2), 4.79, 4.77 (2 d, J = –12.5 Hz each, 2 ϫ 1 H, (150.90 MHz, CDCl3): δ = 148.3 (C-4, triazole), 138.6–125.9 (30 C,
PhCH2), 4.71, 4.69 (2 d, J = –12.7 Hz each, 2ϫ1 H, PhCH2), 4.65
(d, JCH2aOH,CH2bOH = –13.3 Hz, 1 H, CH2aOH), 4.62 (d, 1 H,
CH2bOH), 4.60 (s, 1 H, 1-HB), 4.50 (ddd, J = –14.4, 4.9, 3.4 Hz, 1
Ar-C), 122.8 (C-5, triazole), 101.5 (PhCH), 101.3 (PhCH), 100.7
(C-1B), 98.0 (C-1A), 78.4 (C-4A), 78.3 (C-4B), 77.5 (C-3B), 75.9 (C-
2B), 75.0 (C-2A), 74.5 (PhCH2), 73.5 (C-3A), 72.2 (PhCH2), 71.5
H, CH2CH2a triazole), 4.47 (s, 1 H, 1-HA), 4.35–4.24 (m, 4 H, 6- (PhCH2), 68.5 (C-6A), 68.4 (C-6B), 67.6 (C-5B), 65.4 (CH2CH2 tri-
HaA, 6-HaB, CH2CH2b triazole, CH2aCH2 triazole), 4.23 (dd, J3B,4B azole), 64.2 (C-5A), 56.4 (CH2OH), 49.8 (CH2CH2 triazole) ppm.
Eur. J. Org. Chem. 0000, 0–0
© 0000 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.eurjoc.org
7

Job/Unit: O20041
/KAP1
Date: 10-04-12 15:13:49
Pages: 13
C. Mukherjee, K. Ranta, J. Savolainen, R. Leino
FULL PAPER
HRMS (ESI): calcd. for C52H55N3O12Na [M + Na]+ 936.3683; Propargyl (2,3-Di-O-benzyl-4,6-O-benzylidene-β-
found 936.3690. osyl)-(1Ǟ2)-3-O-benzyl-4,6-O-benzylidene-β- -mannopyranoside
D-mannopyran-
D
(21): Employing the same method as outlined in General Procedure
A, the glycosylation reaction between glycosyl donor 11 (178 mg,
0.33 mmol) and glycosyl acceptor 20 (100 mg, 0.25 mmol) gave di-
saccharide 21 (190 mg, 90%). [α]2D5 = –99.7 (c = 1, CHCl3). 1H
NMR (600.13 MHz, CDCl3): δ = 7.56–7.24 (m, 25 H, Ar-H), 5.61
(s, 1 H, PhCH), 5.43 (s, 1 H, PhCH), 5.07, 4.98 (2 d, J = –12.2 Hz
each, 2ϫ1 H, PhCH2), 4.80 (d, J1B,2B = 0.1 Hz, 1 H, 1-HB), 4.79,
4.77 (2 d, J = –12.6 Hz, 2ϫ1 H, PhCH2), 4.73 (s, 1 H, 1-HA), 4.64,
4.58 (2 d, J = –12.6 Hz, 2ϫ1 H, PhCH2), 4.37 (dd, JOCH2a,OCH2b
= –15.8, JOCH2a,ϵCH = 2.5 Hz, 1 H, OCH2a), 4.34 (dd, JOCH2b,ϵCH
= 2.3 Hz, 1 H, OCH2b), 4.32 (dd, J6aB,6bB = –10.3, J5B,6aB = 4.8 Hz,
1 H, 6-HaB), 4.29 (dd, J6aA,6bA = –10.4, J5A,6aA = 4.8 Hz, 1 H, 6-
HaA), 4.26 (dd, J2A,3A = 3.2 Hz, 1 H, 2-HA), 4.23 (dd, J3B,4B = 9.9,
J4B,5B = 9.2 Hz, 1 H, 4-HB), 4.16 (dd, J2B,3B = 3.2 Hz, 1 H, 2-HB),
4.06 (dd, J3A,4A = 9.9, J4A,5A = 9.3 Hz, 1 H, 4-HA), 3.91 (dd, J5B,6bB
= 10.2 Hz, 1 H, 6-HbB), 3.73 (dd, J5A,6bA = 10.0 Hz, 1 H, 6-HbA),
3.66 (dd, 1 H, 3-HA), 3.59 (dd, 1 H, 3-HB), 3.38 (ddd, 1 H, 5-HA),
3.35 (ddd, 1 H, 5-HB), 2.43 (dd, 1 H, CϵCH) ppm. 13C NMR
(150.90 MHz, CDCl3): δ = 139.0–126.0 (Ar-C), 103.6 (C-1B), 101.6
(PhCH), 101.3 (PhCH), 98.7 (C-1A), 78.3 (C-4B), 78.2 (CϵCH),
78.2 (C-4A), 77.5 (C-3B), 76.7 (C-2A), 76.0 (C-3A), 75.7 (CϵC-H),
75.5 (C-2B), 74.7 (PhCH2), 71.9 (PhCH2), 71.1 (PhCH2), 68.7 (C-
6B), 68.6 (C-6A), 67.6 (C-5B), 67.5 (C-5A), 55.6 (OCH2) ppm.
HRMS (ESI): calcd. for C50H50O11Na [M + Na]+ 849.3251; found
849.3237.
1-[(β- -Mannopyranosyl)-(1Ǟ2)-(α-D-mannopyranosyloxyethyl)]-
D
4-hydroxymethyl-1,2,3-triazole (3): Hydrogenolysis of the coupling
product 18 (55 mg, 0.06 mmol) was carried out following the same
method as outlined in General Procedure C to afford title com-
pound 3 (23 mg, 82%). [α]2D5 = –31.2 (c = 1, H2O). 1H NMR
(600.13 MHz, CD3OD): δ = 7.97 (s, 1 H, triazole-H), 4.87 (br. s, 1
H, 1-HA), 4.69 (br. s, 2 H, CH2abOH), 4.67–4.61 (m, 3 H,
CH2CH2ab triazole, 1-HB), 4.12 (ddd, J = –10.9, 6.8, 3.9 Hz, 1 H,
CH2aCH2 triazole), 4.00 (dd, J2A,3A = 2.6, J1A,2A = 1.8 Hz, 1 H, 2-
HA), 3.93–3.87 (m, 3 H, 2-HB, 6-HaB, CH2bCH2 triazole), 3.75 (dd,
J6aA,6bA = –11.9, J5A,6aA = 2.2 Hz, 1 H, 6-HaA), 3.69 (dd, J6aB,6bB
= –12.0, J5B,6bB = 6.5 Hz, 1 H, 6-HbB), 3.65 (dd, J5A,6bA = 5.7 Hz,
1 H, 6-HbA), 3.63–3.58 (m, 2 H, 3-HA, 4-HA), 3.54 (dd, J3B,4B
≈
J4B,5B ≈ 9.5 Hz, 1 H, 4-HB), 3.46 (dd, J2B,3B = 3.1 Hz, 1 H, 3-HB),
3.25 (ddd, J5B,6aB = 2.2 Hz, 1 H, 5-HB), 3.13 (ddd, 1 H, 5-HA) ppm.
13C NMR (150.90 MHz, CD3OD): δ = 149.3 (C-4, triazole), 125.2
(C-5, triazole), 100.0 (C-1B), 99.1 (C-1A), 78.7 (C-5B), 78.3 (C-2A),
75.2 (C-5A), 75.1 (C-3B), 72.7 (C-2B), 72.0 (C-3A), 68.8 (C-4A), 68.6
(C-4B), 66.9 (CH2CH2 triazole), 63.0 (C-6B), 62.6 (C-6A), 56.6
(CH2OH), 51.3 (CH2CH2 triazole) ppm. HRMS (ESI): calcd. for
C17H29N3O12Na [M + Na]+ 490.1649; found 490.1653.
Propargyl 3-O-Benzyl-4,6-O-benzylidene-2-O-(p-methoxybenzyl)-β-
D-mannopyranoside (19): Employing the same method as outlined
in General Procedure A, the glycosidation reaction between gly-
cosyl donor 7 (700 mg, 1.23 mmol) and propargyl alcohol (83 mg,
86 μL, 1.48 mmol) gave compound 19 (400 mg, 63%). [α]2D5 = –82.8
(c = 1, CHCl3). 1H NMR (600.13 MHz, CDCl3): δ = 7.50–7.48 (m,
2 H, Ar-H), 7.39–7.25 (m, 10 H, Ar-H), 6.85–6.83 (m, 2 H, Ar-H),
5.61 (s, 1 H, PhCH), 4.89, 4.79 (2 d, J = –11.8 Hz each, 2ϫ1 H,
PhCH2), 4.73 (d, J1,2 = 1.0 Hz, 1 H, 1-H), 4.66, 4.56 (2 d, J =
–12.5 Hz each, 2ϫ1 H, PhCH2), 4.43 (dd, JOCH2a,OCH2b = –15.9,
JOCH2a,ϵCH = 2.4 Hz, 1 H, OCH2a), 4.40 (dd, JOCH2b,ϵCH = 2.4 Hz,
1 H, OCH2b), 4.31 (dd, J6a,6b = –10.4, J5,6a = 4.8 Hz, 1 H, 6-Ha),
4.19 (dd, J3,4 = 9.9, J4,5 = 9.3 Hz, 1 H, 4-H), 3.95 (dd, J2,3 = 3.1 Hz,
1 H, 2-H), 3.92 (dd, J5,6b = 10.1 Hz, 1 H, 6-Hb), 3.80 (s, 3 H,
OCH3), 3.61 (dd, 1 H, 3-H), 3.36 (ddd, 1 H, 5-H), 2.45 (dd, 1 H,
CϵCH) ppm. 13C NMR (150.90 MHz, CDCl3): δ = 159.2 (Ar-C),
138.3–126.0 (Ar-C), 113.5 (Ar-C), 101.4 (PhCH), 99.3 (C-1), 78.6
(CϵCH), 78.5 (C-4), 77.8 (C-3), 75.3 (C-2), 75.2 (CϵCH), 74.5
(PhCH2), 72.3 (PhCH2), 68.5 (C-6), 67.6 (C-5), 55.7 (OCH2), 55.2
(OCH3) ppm. HRMS (ESI): calcd. for C31H32O7Na [M + Na]+
539.2046; found 539.2032.
Propargyl 2-O-Acetyl-3-O-benzyl-4,6-O-benzylidene-α-D-mannopyr-
anoside (22): The glycosidation reaction was carried out starting
from glycosyl donor 8 (1.0 g, 2.03 mmol) and propargyl alcohol
(137 mg, 142 μL, 2.44 mmol) following the same reaction pro-
cedure as described for the preparation of compound 13 providing
the title compound 22 (730 mg, 82%). [α]2D5 = +23.7 (c = 1, CHCl3).
1H NMR (600.13 MHz, CDCl3): δ = 7.51–7.24 (m, 10 H, Ar-H),
5.63 (s, 1 H, PhCH), 5.43 (dd, J2,3 = 3.5, J1,2 = 1.6 Hz, 1 H, 2-H),
4.99 (d, 1 H, 1-H), 4.70, 4.65 (2 d, J = –12.2 Hz each, 2ϫ1 H,
PhCH2), 4.27 (dd, J6a,6b = –10.3, J5,6a = 4.8 Hz, 1 H, 6-Ha), 4.24
(dd, JOCH2a,OCH2b = –17.0, JOCH2a,ϵCH = 2.4 Hz, 1 H, OCH2a),
4.24 (dd, JOCH2b,ϵCH = 2.4 Hz, 1 H, OCH2b), 4.07 (dd, J3,4 = 9.9,
J4,5 = 9.5 Hz, 1 H, 4-H), 4.01 (dd, 1 H, 3-H), 3.88 (ddd, J5,6b
=
10.4 Hz, 1 H, 5-H), 3.85 (dd, 1 H, 6-Hb), 2.46 (dd, 1 H, CϵCH),
2.17 (s, 3 H, COCH3) ppm. 13C NMR (150.90 MHz, CDCl3): δ =
170.1 (COCH3), 137.9–126.1 (Ar-C), 101.6 (PhCH), 97.5 (C-1),
78.2 (CϵC-H), 78.1 (C-4), 75.3 (CϵC-H), 73.7 (C-3), 72.1
(PhCH2), 69.4 (C-2), 68.5 (C-6), 64.3 (C-5), 54.7 (OCH2), 21.0
(COCH3) ppm. HRMS (ESI): calcd. for C25H26O7Na [M + Na]+
461.1576; found 461.1550.
Propargyl 3-O-Benzyl-4,6-O-benzylidene-β-D-mannopyranoside (20):
Starting from compound 19 (400 mg, 0.77 mmol), the p-meth-
oxybenzyl deprotection method was the same as that used for the
preparation of compound 10 providing the title compound 20
Propargyl 3-O-Benzyl-4,6-O-benzylidene-α-D-mannopyranoside
(23): Compound 22 (400 mg, 0.91 mmol) was deacetylated follow-
ing the same method as described for the preparation of compound
14 to give the title compound 23 (330 mg, 91%). [α]2D5 = +67.7 (c
= 1, CHCl3). 1H NMR (600.13 MHz, CDCl3): δ = 7.50–7.28 (m,
10 H, Ar-H), 5.61 (s, 1 H, PhCH), 5.08 (d, J1,2 = 1.4 Hz, 1 H, 1-
H), 4.85, 4.71 (2 d, J = –11.8 Hz each, 2ϫ1 H, PhCH2), 4.27 (dd,
(240 mg, 78%). [α]2D5
= –62.6 (c =
1, CHCl3). 1H NMR
(600.13 MHz, CDCl3): δ = 7.51–7.25 (m, 10 H, Ar-H), 5.61 (s, 1
H, PhCH), 4.86, 4.78 (2 d, J = –12.2 Hz each, 2ϫ1 H, PhCH2),
4.79 (d, J1,2 = 1.2 Hz, 1 H, 1-H), 4.44 (dd, JOCH2a,OCH2b = –17.1,
JOCH2a,ϵCH = 2.4 Hz, 1 H, OCH2a), 4.43 (dd, JOCH2b,ϵCH = 2.4 Hz,
1 H, OCH2b), 4.34 (dd, J6a,6b = –10.5, J5,6a = 4.9 Hz, 1 H, 6-Ha),
J6a,6b = –10.6, J5,6a = 5.3 Hz, 1 H, 6-Ha), 4.25 (dd, JOCH2a,OCH2b
–17.0, JOCH2a,ϵCH = 2.4 Hz, 1 H, OCH2a), 4.25 (dd, JOCH2b,ϵCH
=
=
4.16 (ddd, J2,3 = 3.3, J2,2-OH = 1.5 Hz, 1 H, 2-H), 4.14 (dd, J3,4
=
9.5, J4,5 = 9.4 Hz, 1 H, 4-H), 3.88 (dd, J5,6b = 10.1 Hz, 1 H, 6-Hb),
3.70 (dd, 1 H, 3-H), 3.39 (ddd, 1 H, 5-H), 2.56 (d, 1 H, 2-OH),
2.4 Hz, 1 H, OCH2b), 4.12 (dd, J3,4 = 9.6, J4,5 = 9.4 Hz, 1 H, 4-H),
4.09 (ddd, J2,3 = 3.5, J2,2-OH = 1.4 Hz, 1 H, 2-H), 3.93 (dd, 1 H, 3-
2.46 (dd, 1 H, CϵCH) ppm. 13C NMR (150.90 MHz, CDCl3): δ H), 3.86 (ddd, J5,6b = 10.3 Hz, 1 H, 5-H), 3.85 (dd, 1 H, 6-Hb),
= 137.8–126.0 (Ar-C), 101.6 (PhCH), 97.5 (C-1), 78.3 (C-2), 78.2
(CϵCH), 76.7 (C-3), 75.5 (CϵCH), 72.6 (PhCH2), 69.8 (C-4), 68.5
(C-6), 66.9 (C-5), 55.7 (OCH2) ppm. HRMS (ESI): calcd. for
C23H24O6Na [M + Na]+ 419.1471; found 419.1462.
2.70 (d, 1 H, 2-OH), 2.46 (dd, 1 H, CϵCH) ppm. 13C NMR
(150.90 MHz, CDCl3): δ = 137.9–126.0 (Ar-C), 101.6 (PhCH), 98.6
(C-1), 78.7 (C-4), 78.5 (CϵCH), 75.4 (C-3), 75.0 (CϵCH), 73.0
(PhCH2), 69.8 (C-2), 68.7 (C-6), 63.7 (C-5), 54.5 (OCH2) ppm.
8
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© 0000 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Eur. J. Org. Chem. 0000, 0–0

Job/Unit: O20041
/KAP1
Date: 10-04-12 15:13:49
Pages: 13
β-Linked Mono- and Divalent Mannosides
HRMS (ESI): calcd. for C23H24O6Na [M + Na]+ 419.1471; found
419.1460.
as outlined in General Procedure C to give compound 4 (21 mg,
86%). [α]2D5 = –43.0 (c = 1, H2O). 1H NMR (600.13 MHz, CD3OD):
δ = 8.16 (s, 1 H, triazole-H), 4.95 (d, JOCH2a triazole,OCH2b triazole
=
Propargyl (2,3-Di-O-benzyl-4,6-O-benzylidene-β-
D-mannopyran-
–12.5 Hz, 1 H, OCH2a triazole), 4.81 (d, 1 H, OCH2b triazole),
4.68–4.62 (m, 2 H, CH2CH2ab triazole), 4.61 (d, J1A,2A = 0.8 Hz, 1
H, 1-HA), 4.49 (d, J1B,2B = 0.7 Hz, 1 H, 1-HB), 4.26 (ddd, J = –11.4,
4.9, 4.9 Hz, 1 H, CH2aCH2 triazole), 4.00 (ddd, J = 6.8, 4.4 Hz, 1
H, CH2bCH2 triazole), 3.91 (dd, J6aA.6bA = –11.8, J5A,6aA = 2.3 Hz,
1 H, 6-HaA), 3.87 (dd, J6aB.6bB = –11.8, J5B,6aB = 2.3 Hz, 1 H, 6-
osyl)-(1Ǟ2)-3-O-benzyl-4,6-O-benzylidene-α- -mannopyranoside
D
(24): Employing the same method as outlined in General Procedure
A, the glycosylation reaction between glycosyl donor 11 (177 mg,
0.33 mmol) and glycosyl acceptor 23 (100 mg, 0.25 mmol) gave di-
saccharide 24 (134 mg, 64%). [α]2D5 = –34.4 (c = 1, CHCl3). 1H
NMR (600.13 MHz, CDCl3): δ = 7.53–7.20 (m, 25 H, Ar-H), 5.60
(s, 1 H, PhCH), 5.50 (s, 1 H, PhCH), 5.05, 4.97 (2 d, J = –12.2 Hz
each, 2ϫ1 H, PhCH2), 5.04 (d, J1A,2A = 1.4 Hz, 1 H, 1-HA), 4.77,
4.72 (2 d, J = –12.2 Hz each, 2ϫ1 H, PhCH2), 4.70, 4.62 (2 d, J
= –12.5 Hz each, 2ϫ1 H, PhCH2), 4.64 (s, 1 H, 1-HB), 4.28 (dd,
H
aB), 3.85 (dd, J2A,3A = 3.2 Hz, 1 H, 2-HA), 3.82 (d, J2B,3B
3.2 Hz, 1 H, 2-HB), 3.74 (dd, J5A,6bA = 6.1 Hz, 1 H, 6-HbA), 3.69
(dd, J5B,6bB = 6.1 Hz, 1 H, 6-HbB), 3.56 (dd, J3A,4A ≈ J4A,5A
=
≈
9.5 Hz, 1 H, 4-HA), 3.53 (dd, J3B,4B ≈ J4B,5B ≈ 9.5 Hz, 1 H, 4-HB),
3.46 (dd, 1 H, 3-HA), 3.43 (dd, 1 H, 3-HB), 3.26 (ddd, 1 H, 5-HA),
3.21 (ddd, 1 H, 5-HB) ppm. 13C NMR (150.90 MHz, CD3OD): δ
= 145.3 (C-4, triazole), 126.7 (C-5, triazole), 101.9 (C-1B), 100.5
(C-1A), 78.5 (C-5B), 78.4 (C-5A), 75.2 (C-3A), 75.1 (C-3B), 72.4 (C-
2A), 72.3 (C-2B), 68.8 (CH2CH2 triazole), 68.6 (C-4A), 68.5 (C-4B),
62.9 (C-6A), 62.8 (C-6B), 62.4 (OCH2 triazole), 51.7 (CH2CH2 tri-
azole) ppm. HRMS (ESI): calcd. for C17H28N3O12 [M – H]+
466.1673; found 466.1713.
J2A,3A = 3.2 Hz, 1 H, 2-HA), 4.26 (dd, J6aA,6bA = –10.6, J5A,6aA
=
5.0 Hz, 1 H, 6-HaA), 4.26 (dd, J4B,5B = 10.2, J3B,4B = 9.9 Hz, 1 H,
4-HB), 4.25 (dd, J6aB,6bB = –10.5, J5B,6aB = 4.8 Hz, 1 H, 6-HaB),
4.24 (dd, JOCH2a,OCH2b = –17.2, JOCH2a,ϵCH = 2.5 Hz, 1 H,
OCH2a), 4.23 (dd, JOCH2b,ϵCH = 2.3 Hz, 1 H, OCH2b), 4.12 (dd,
J3A,4A = 9.9, J4A,5A = 9.7 Hz, 1 H, 4-HA), 4.00 (dd, J2B,3B = 3.2 Hz,
1 H, 2-HB), 3.96 (dd, 1 H, 3-HA), 3.87 (dd, J5B,6bB = 9.9 Hz, 1 H,
6-HbB), 3.82 (ddd, J5A,6bA = 10.0 Hz, 1 H, 5-HA), 3.77 (dd, 1 H, 6-
1-[(2,3-Di-O-benzyl-4,6-O-benzylidene-β-
(1Ǟ2)-(3-O-benzyl-4,6-O-benzylidene-β- -mannopyranosyloxy-
ethyl)]-4-[(2,3-di-O-benzyl-4,6-O-benzylidene-β- -mannopyranos-
yl)-(1Ǟ2)-(3-O-benzyl-4,6-O-benzylidene-β- -mannopyranosyloxy-
D-mannopyranosyl)-
H
bA), 3.60 (dd, 1 H, 3-HB), 3.32 (ddd, 1 H, 5-HB), 2.46 (dd, 1 H,
CϵCH) ppm. 13C NMR (150.90 MHz, CDCl3): δ = 138.7–126.0
(Ar-C), 101.6 (PhCH), 101.4 (PhCH), 100.7 (C-1B), 96.8 (C-1A),
78.4 (2 C, C-4A, C-4B), 78.3 (CϵCH), 77.6 (C-3B), 75.9 (C-2B), 75.2
(C-2A), 74.7 (CϵCH), 74.6 (PhCH2), 74.0 (C-3A), 72.2 (PhCH2),
71.4 (PhCH2), 68.7 (C-6B), 68.5 (C-6A), 67.8 (C-5B), 64.6 (C-5A),
54.4 (OCH2) ppm. HRMS (ESI): calcd. for C50H54NO11 [M +
NH4]+ 844.3697; found 844.3681.
D
D
D
methyl)]-1,2,3-triazole (26): Employing the same method as out-
lined in General Procedure B, the click coupling reaction between
2-azidoethyl glycoside 12 (52 mg, 0.06 mmol) and propargyl glycos-
ide 21 (50 mg, 0.06 mmol) gave compound 26 (88 mg, 86%). [α]2D5
= –87.9 (c = 1, CHCl3). 1H NMR (600.13 MHz, CDCl3): δ = 7.53–
7.16 (m, 50 H, Ar-H), 7.04 (s, 1 H, triazole-H), 5.58 (s, 1 H, PhCH),
5.56 (s, 1 H, PhCH), 5.44 (s, 1 H, PhCH), 5.42 (s, 1 H, PhCH),
5.04, 4.94 (2 d, J = –12.5 Hz each, 2ϫ1 H, PhCH2), 5.01, 4.91 (2
d, J = –12.6 Hz each, 2ϫ1 H, PhCH2), 4.82 (br. s, 1 H, 1-HB),
4.80 (d, J = –12.7 Hz, 1 H, PhCH2), 4.77–4.72 (m, 4 H, OCH2a
triazole, PhCH2), 4.70, 4.57 (2 d, J = –12.5 Hz each, 2 ϫ 1 H,
PhCH2), 4.68, 4.54 (2 d, J = –12.1 Hz each, 2ϫ1 H, PhCH2), 4.53
(br. s, 1 H, 1-HD), 4.50 (d, JOCH2a triazole,OCH2b triazole = –12.7 Hz, 1
H, OCH2b triazole), 4.33 (br. s, 1 H, 1-HA), 4.29 (br. s, 1 H, 1-HC),
4.29–4.17 (m, 9 H, 6-HaA, 6-HaB, 6-HaC, 6-HaD, 4-HB, 4-HD, 2-HA,
CH2CH2ab triazole), 4.16 (d, J2C,3C = 3.1 Hz, 1 H, 2-HC), 4.10 (d,
J2B,3B = 3.1 Hz, 1 H, 2-HB), 4.07–4.02 (m, 1 H, CH2aCH2 triazole),
4.00 (dd, J3A,4A = 9.7, J4A,5A = 9.6 Hz, 1 H, 4-HA), 3.95 (dd, J3C,4C
= 9.7, J4C,5C = 9.6 Hz, 1 H, 4-HC), 3.92 (d, J2D,3D = 3.0 Hz, 1 H,
2-HD), 3.90–3.85 (m, 2 H, 6-HbB, 6-HbD), 3.82 (ddd, J = –11.0, 8.1,
3.9 Hz, 1 H, CH2bCH2 triazole), 3.73 (dd, J6aA,6bA = –10.1, J5A,6bA
= 5.5 Hz, 1 H, 6-HbA), 3.71 (dd, J6aC,6bC = –10.2, J5C,6bC = 5.4 Hz,
1 H, 6-HbC), 3.61 (dd, J3D,4D = 10.0 Hz, 1 H, 3-HD), 3.50 (dd,
J3B,4B = 9.8 Hz, 1 H, 3-HB), 3.49–3.45 (m, 2 H, 3-HA, 3-HC), 3.21–
3.07 (m, 4 H, 5-HA , 5-HB, 5-HC, 5-HD ) ppm. 1 3 C NMR
(150.90 MHz, CDCl3): δ = 144.0 (C-4, triazole), 139.1–125.9 (60
C, Ar-C), 122.3 (C-5, triazole), 103.2 (C-1B), 102.9 (C-1D), 101.6
(PhCH), 101.5 (PhCH), 101.4 (C-1C), 101.2 (PhCH), 101.1
(PhCH), 100.6 (C-1A), 78.6 (C-4B), 78.2 (C-4D), 78.1 (C-4A), 77.9
(C-4C), 77.8 (C-3B), 77.6 (C-3D), 75.9 (C-2D), 75.8 (C-2A), 75.6 (C-
3C), 75.5 (C-2B), 75.4 (C-3A), 74.9 (C-2C), 74.4 (PhCH2), 74.3
(PhCH2), 72.2 (PhCH2), 71.6 (PhCH2), 70.9 (PhCH2), 70.8
(PhCH2), 68.6 (2 C, C-6A, C-6C), 68.5 (2 C, C-6B, C-6D), 67.7 (C-
5A), 67.6 (C-5C), 67.5 (C-5B), 67.4 (C-5D), 67.1 (CH2CH2 triazole),
62.0 (OCH2 triazole), 49.9 (CH2CH2 triazole) ppm. HRMS (ESI):
calcd. for C99H102N3O22 [M + H]+ 1684.6955; found 1684.6948.
1-(3-O-Benzyl-4,6-O-benzylidene-β-
(3-O-benzyl-4,6-O-benzylidene-β-
D
-mannopyranosyloxyethyl)-4-
D
-mannopyranosyloxymethyl)-
1,2,3-triazole (25): Employing the same method as outlined in Ge-
neral Procedure B, the click coupling reaction between 2-azidoethyl
glycoside 10 (190 mg, 0.44 mmol) and propargyl glycoside 20
(175 mg, 0.44 mmol) gave compound 25 (300 mg, 83%). [α]2D5
=
–44.3 (c = 1, CHCl3). 1H NMR (600.13 MHz, CDCl3): δ = 7.77 (s,
1 H, triazole-H), 7.50–7.25 (m, 20 H, Ar-H), 5.59 (s, 1 H, PhCH),
5.57 (s, 1 H, PhCH), 4.98 (d, JOCH2a triazole,OCH2b triazole = –12.4 Hz,
1 H, OCH2a triazole), 4.82, 4.74 (2 d, J = –12.3 Hz each, 2ϫ1 H,
PhCH2), 4.81, 4.72 (2 d, J = –12.2 Hz each, 2ϫ1 H, PhCH2), 4.80
(d, 1 H, OCH2b triazole), 4.68 (br. s, 1 H, 1-HA), 4.62–4.52 (m, 2
H, CH2CH2ab triazole), 4.46 (br. s, 1 H, 1-HB), 4.36 (dd, J6aA,6bA
= –10.4, J5A,6aA = 4.9 Hz, 1 H, 6-HaA), 4.29 (dd, J6aB,6bB = –10.4,
J5B,6aB = 4.9 Hz, 1 H, 6-HaB), 4.25 (ddd, J = –10.8, 3.9, 3.9 Hz, 1
H, CH2aCH2 triazole), 4.13 (dd, J3A,4A ≈ J4A,5A ≈ 9.5 Hz, 1 H, 4-
HA), 4.09 (dd, J3B,4B = 9.5, J4B,5B = 9.4 Hz, 1 H, 4-HB), 4.09 (ddd,
J2A,3A = 3.2 Hz, 1 H, 2-HA), 4.05 (ddd, J2B,3B = 3.3, J2B,2-OH
=
1.3 Hz, 1 H, 2-HB), 3.96 (ddd, J = 8.0, 3.8 Hz, 1 H, CH2bCH2
triazole), 3.89 (dd, J5A,6bA = 10.3 Hz, 1 H, 6-HbA), 3.84 (dd, J5B,6bB
= 10.3 Hz, 1 H, 6-HbB), 3.64 (dd, 1 H, 3-HA), 3.61 (dd, 1 H, 3-
HB), 3.40 (ddd, 1 H, 5-HA), 3.31 (ddd, 1 H, 5-HB), 2.79 (d, 1 H,
2-OHB), 2.77 (br. s, 1 H, 2-OHA) ppm. 13C NMR (150.90 MHz,
CDCl3): δ = 143.7 (C-4, triazole), 137.8–125.9 (24 C, Ar-C), 124.5
(C-5, triazole), 101.5 (PhCH), 101.4 (PhCH), 100.6 (C-1B), 99.2 (C-
1A), 78.3 (C-4A), 78.2 (C-4B), 76.6 (C-3A), 76.4 (C-3B), 72.5
(PhCH2), 72.4 (PhCH2), 69.7 (C-2A), 69.5 (C-2B), 68.5 (C-6A), 68.3
(C-6B), 67.9 (OCH2CH2), 66.9 (C-5B), 66.8 (C-5A), 62.2 (OCH2 tri-
azole), 50.2 (CH2CH2 triazole) ppm. HRMS (ESI): calcd. for
C45H49N3O12Na [M + Na]+ 846.3214; found 846.3214.
1-(β-D-Mannopyranosyloxyethyl)-4-(β-D-mannopyranosyloxy-
methyl)-1,2,3-triazole (4): Hydrogenolysis of the coupling product
25 (43 mg, 0.05 mmol) was carried out following the same method
1-[(β-
D
-Mannopyranosyl)-(1Ǟ2)-(β-
D
-mannopyranosyloxyethyl)]-4-
[(β- -mannopyranosyl)-(1Ǟ2)-[(β-
D
D-mannopyranosyloxymethyl)]-
Eur. J. Org. Chem. 0000, 0–0
© 0000 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.eurjoc.org
9

Job/Unit: O20041
/KAP1
Date: 10-04-12 15:13:49
Pages: 13
C. Mukherjee, K. Ranta, J. Savolainen, R. Leino
FULL PAPER
1,2,3-triazole (5): Hydrogenolysis of the coupling product 26
(44 mg, 0.03 mmol) was carried out following the same method as
outlined in General Procedure C to give compound 5 (20 mg, 97%).
1-[(β-
D
-Mannopyranosyl)-(1Ǟ2)-(α-
D
-mannopyranosyloxyethyl)]-4-
[(β- -mannopyranosyl)-(1Ǟ2)-(α-
D
D-mannopyranosyloxymethyl)]-
1,2,3-triazole (6): Hydrogenolysis of the coupling product 27
(70 mg, 0.04 mmol) was carried out following the same method as
outlined in General Procedure C to give compound 6 (25 mg, 76%).
[α]2D5 = –39.9 (c = 1, H2O). H NMR (600.13 MHz, CD3OD): δ =
1
8.10 (s, 1 H, triazole-H), 5.00 (d, JOCH2a triazole,OCH2b triazole
=
1
–12.5 Hz, 1 H, OCH2a triazole), 4.83 (d, 1 H, OCH2b triazole), 4.76
(br. s, 1 H, 1-HB), 4.75 (br. s 1 H, 1-HA), 4.69–4.63 (m, 2 H,
CH2CH2ab triazole), 4.65 (br. s, 1 H, 1-HC), 4.60 (br. s, 1 H, 1-HD),
4.31 (ddd, J = –11.2, 6.2, 3.7 Hz, 1 H, CH2aCH2 triazole), 4.17 (d,
J2A,3A = 3.2 Hz, 1 H, 2-HA), 4.07 (d, J2C,3C = 3.3 Hz, 1 H, 2-HC),
4.00 (ddd, J = 6.7, 3.5 Hz, 1 H, CH2bCH2 triazole), 3.96 (d, J2B,3B
= 3.1 Hz, 1 H, 2-HB), 3.93 (dd, J6aA,6bA = –11.9, J5A,6aA = 2.2 Hz,
[α]2D5 = +1.4 (c = 1, H2O). H NMR (600.13 MHz, CD3OD): δ =
8.07 (s, 1 H, triazole-H), 5.05 (d, J1A,2A = 1.5 Hz, 1 H, 1-HA), 4.87
(d, J1C,2C = 1.6 Hz, 1 H, 1-HC), 4.83 (d, JOCH2a triazole,OCH2b triazole
= –12.4 Hz, 1 H, OCH2a triazole), 4.69 (br. s, 1 H, 1-HB), 4.67 (d,
1 H, OCH2b triazole), 4.68–4.64 (m, 2 H, CH2CH2ab triazole), 4.64
(br. s, 1 H, 1-HD), 4.13 (ddd, J = –10.9, 6.7, 3.8 Hz, 1 H, CH2aCH2
triazole), 4.09 (dd, J2A,3A = 3.4 Hz, 1 H, 2-HA), 4.00 (dd, J2C,3C
=
1 H, 6-HaA), 3.90–3.84 (m, 3 H, 6-HaC, 6-HaB, 6-HaD), 3.77 (d, 2.9 Hz, 1 H, 2-HC), 3.94–3.90 (m, 2 H, 2-HB, CH2bCH2 triazole),
J2D,3D = 3.1 Hz, 1 H, 2-HD), 3.74 (dd, J5A,6bA = 6.1 Hz, 1 H, 6- 3.89–3.83 (m, 4 H, 2-HD, 6-HaD, 6-HaB, 6-HaA), 3.77–3.59 (m, 9 H,
HbA), 3.71 (dd, J6aC,6bC = –11.9, J5C,6bC = 6.1 Hz, 1 H, 6-HbC),
3.69–3.66 (m, 2 H, 6-HbB, 6-HbD), 3.55 (dd, J3A,4A = 9.5, J4A,5A
6-HaC, 3-HA, 6-HbA, 6-HbC, 6-HbB, 6-HbD, 4-HA, 3-HC, 4-HC),
3.58–3.56 (m, 1 H, 5-HA), 3.55–3.51 (m, 2 H, 4-HB, 4-HD), 3.48
=
9.4 Hz, 1 H, 4-HA), 3.53–3.42 (m, 6 H, 4-HC, 4-HD, 4-HB, 3-HA, (dd, J3B,4B = 8.2, J2B,3B = 3.1 Hz, 1 H, 3-HB), 3.47 (dd, J3D,4D
=
3-HC, 3-HD), 3.38 (dd, J3B,4B = 9.4 Hz, 1 H, 3-HB), 3.33–3.27 (m, 8.2, J2D,3D = 3.1 Hz, 1 H, 3-HD), 3.28–3.24 (m, 2 H, 5-HB, 5-HD),
1 H, 5-HA), 3.24–3.14 (m, 3 H, 5-HC, 5-HB, 5-HD) ppm. 13C NMR 3.20 (ddd, J6aC,6bC = –8.4, J5C,6bC = 5.6, J5C,6aC = 2.1 Hz, 1 H, 5-
(150.90 MHz, CDCl3): δ = 145.5 (C-4, triazole), 125.8 (C-5, tri-
HC) ppm. 13C NMR (150.90 MHz, CD3OD): δ = 145.4 (C-4, tri-
azole), 102.2 (C-1C), 102.1 (C-1B), 101.8 (C-1D), 101.2 (C-1A), 79.1
azole), 126.0 (C-5, triazole), 99.8 (2 C, C-1B, C-1D), 98.9 (C-1C),
(C-2A), 78.6 (C-5A), 78.5 (2 C, C-5B, C-5C), 78.4 (C-5D), 78.3 (C- 98.6 (C-1A), 78.5 (2 C, C-5B, C-5D), 78.3 (C-2A), 78.1 (C-2C), 75.0
2C), 75.0 (C-3B), 74.9 (C-3D), 74.5 (C-3A), 74.3 (C-3C), 72.1 (C-2B),
72.0 (C-2D), 69.0 (CH2CH2 triazole), 68.9 (C-4A), 68.8 (C-4C), 68.6
(C-4B), 68.5 (C-4D), 63.0 (OCH2 triazole), 62.9 (C-6A), 62.8 (C-
6C), 62.7 (C-6B), 62.6 (C-6D), 51.9 (CH2CH2 triazole) ppm. HRMS
(ESI): calcd. for C29H49N3O22Na [M + Na]+ 814.2705; found
814.2698.
(4 C, C-5A, C-3B, C-5C, C-3D), 75. 2 (2 C, C-2B, C-2D), 71.8 (2 C,
C-3A, C-3C), 68.9 (C-4A), 68.6 (C-4C), 68.4 (2 C, C-4B, C-4D), 66.7
(CH2CH2 triazole), 62.8 (2 C, C-6A, C-6C), 62.6 (C-6B), 62.5 (C-
6D), 61.1 (OCH2 triazole), 51.3 (CH2CH2 triazole) ppm. HRMS
(ESI): calcd. for C29H48N3O22 [M – H]+ 790.2730; found 790.2796.
Biological Studies
1-[(2,3-Di-O-benzyl-4,6-O-benzylidene-β-
(1Ǟ2)-(3-O-benzyl-4,6-O-benzylidene-α- -mannopyranosyloxy-
ethyl)]-4-[(2,3-di-O-benzyl-4,6-O-benzylidene-β- -mannopyranos-
yl)-(1Ǟ2)-(3-O-benzyl-4,6-O-benzylidene-α- -mannopyranosyloxy-
D-mannopyranosyl)-
Synthesis of <3 kDa Candida albicans Mannan: C. albicans mannan
(20 mg/mL) was hydrolyzed under mild acidic conditions with 0.1 n
HCl for up to 60 min at 100 °C. The hydrolysis products were neu-
tralized with NaOH and analyzed by TLC using silica gel coated
aluminium sheets (Merck, Darmstadt, Germany) using n-butanol/
acetic acid/water (2:1:1, v/v/v) as eluent. The hydrolysis products
were detected on TLC by using orcinol (2 g/L in 20% sulfuric acid)
by heating the sheets for 10 min at 105 °C. Fractionation of the
hydrolysis products by size was performed with Amicon YM3 YM1
ultrafiltration membranes (Millipore, Bedford, MA, USA) accord-
ing to the manufacturer’s instructions. The final fraction of Ͻ3 kDa
was sterile-filtered and lyophilized for storage at 4 °C.
D
D
D
methyl)]-1,2,3-triazole (27): Employing the same method as out-
lined in General Procedure B, the click coupling reaction between
2-azidoethyl glycoside 15 (52 mg, 0.06 mmol) and propargyl glycos-
ide 24 (50 mg, 0.06 mmol) gave compound 27 (90 mg, 88%). [α]2D5
= –40.0 (c = 1, CHCl3). 1H NMR (600.13 MHz, CDCl3): δ = 7.52–
7.20 (m, 51 H, triazole-H, Ar-H), 5.59 (s, 1 H, PhCH), 5.58 (s, 1
H, PhCH), 5.50 (s, 1 H, PhCH), 5.46 (s, 1 H, PhCH), 5.03 (d, J =
–12.2 Hz, 1 H, PhCH2), 4.99 (m, 2 H, 1-HA, PhCH2), 4.95, 4.93 (2
d, J = –12.2 Hz each, 2ϫ1 H, PhCH2), 4.78 (br. s, 1 H, 1-HC),
4.77–4.58 (m, 12 H, 1-HB, 1-HD, OCH2ab triazole, PhCH2), 4.56–
Competitive ELISA Inhibition: Polystyrene 96-well ELISA plates
4.53 (m, 1 H, CH2CH2a triazole), 4.50–4.46 (m, 1 H, CH2CH2b were coated with hydrolyzed low-molecular-weight (Ͻ3 kDa) Can-
triazole), 4.28–4.19 (m, 6 H, 6-HaB, 6-HaA, 2-HA, 4-HB, 4-HD, 6- dida albicans Cetavlon mannan (50 μL, 0.1 mg/mL in PBS) and in-
HaD), 4.16 (dd, J6aC,6bC = –10.3, J5C,6aC = 4.8 Hz, 1 H, 6-HaC), cubated at 4 °C overnight. The plates were then washed three times
4.14–4.06 (m, 4 H, 2-HC, 4-HA, 4-HC, CH2aCH2 triazole), 3.96– with 0.05% PBS-Tween. Blocking was achieved by using 2% BSA-
3.94 (m, 2 H, 2-HB, 2-HD), 3.92 (dd, J3A,4A = 9.9, J2A,3A = 3.3 Hz,
PBS (Bovine albumin fraction V, ICN Biomedicals Inc., Aurora,
1 H, 3-HA), 3.88–3.75 (m, 6 H, CH2bCH2 triazole, 6-HbB, 3-HC, 5- Ohio, 810034; Tween 20 Fluka, Sigma–Aldrich Co. 93773) for 2 h
HA, 6-HbD, 6-HbA), 3.71 (dd, J5C,6bC = 10.3 Hz, 1 H, 6-HbC), 3.60– at room temp. after which the plates were washed three times. The
3.57 (m, 2 H, 3-HB, 3-HD), 3.44 (ddd, 1 H, 5-HC), 3.33 (ddd,
serum samples were collected from vaginal candidiasis patients and
J6aD,6bD = –9.8, J5D,6bD = 9.8, J5D,6aD = 5.0 Hz, 1 H, 5-HD), 3.28 had elevated levels of IgG antibodies against C. albicans mannan
(ddd, J6aB,6bB = –9.7, J5B,6bB = 9.7, J5B,6aB = 4.9 Hz, 1 H, 5- (50 μL, diluted 1:100 in 0.5% BSA/0.05% PBS-Tween) and incu-
HB) ppm. 13C NMR (150.90 MHz, CDCl3): δ = 143.9 (C-4 tri-
azole), 138.8–125.8 (60 C, Ar-C), 123.5 (C-5, triazole), 101.5
(PhCH), 101.4 (PhCH), 101.3 (2 C, PhCH), 100.7 (C-1D), 100.6
(C-1B), 98.3 (C-1C), 97.8 (C-1A), 78.5, 78.4, 78.3 (4 C, C-4A, C-4C,
bated at 37 °C for 2 h with serial ten-fold dilutions of the studied
saccharides (0.0001–1 mg/mL) in a total volume of 100 μL. There-
after the serum dilutions were applied to the coated ELISA plates
and blotted and incubated at 4 °C overnight. After washing three
C-4B, C-4D), 77.6 (2 C, C-3B, C-3D), 76.0 (C-2B), 75.8 (C-2D), 74.9 times, 100 μL alkaline phosphatase conjugated antihuman IgG
(C-2C), 74.8 (C-2A), 74.6 (PhCH2), 74.5 (PhCH2), 74.0 (C-3A), 73.6 (Dako, Denmark, DO 336; 1:1000 in 0.5% BSA/PBS-Tween) was
(C-3C), 72.3 (PhCH2), 72.2 (PhCH2), 71.4 (PhCH2), 71.2 (PhCH2), applied to the plates and they were incubated for 2 h at 37 °C. After
68.8, 68.6, 68.5 (4 C, C-6A, C-6C, C-6B, C-6D), 67.7 (C-5B), 67.6 (C-
5D), 65.5 (CH2CH2 triazole), 64.6 (C-5C), 64.4 (C-5A), 60.3 (OCH2
washing three times, the color reaction was developed by using p-
nitrophenylphosphate disodium salt (100 μL, 1 mg/mL in diethan-
triazole), 49.8 (CH2CH2 triazole) ppm. HRMS (ESI): calcd. for olamine, Reagena, Finland, 180288) as substrate and terminated
C99H105N4O22 [M + NH4]+ 1701.7221; found 1701.7191.
by the addition of 1 m NaOH (AKZO Nobel, The Netherlands)
10
www.eurjoc.org
© 0000 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Eur. J. Org. Chem. 0000, 0–0

Job/Unit: O20041
/KAP1
Date: 10-04-12 15:13:49
Pages: 13
β-Linked Mono- and Divalent Mannosides
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automated microplate reader. The inhibition in ELISA was ex-
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no inhibition – O.D. PBS)%. Mean values of the duplicate determi-
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Acknowledgments
This work was financially supported by grants from the Finnish
Allergy Research Foundation and the Finnish Society of Allergol-
ogy and Immunology (to J. S. and co-workers), and the Academy
of Finland [grant numbers #122126 and #252281 (to J. S. and co-
workers), and #121335 and #252371 (to R. L. and co-workers)].
Dr. Filip S. Ekholm is thanked for the Perch NMR analysis and
fruitful scientific discussions. Markku Reunanen is thanked for the
HRMS analyses and Leena Kavén-Honka for excellent technical
assistance.
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Date: 10-04-12 15:13:49
Pages: 13
C. Mukherjee, K. Ranta, J. Savolainen, R. Leino
FULL PAPER
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Received: January 12, 2012
Published Online: ᭿
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Eur. J. Org. Chem. 0000, 0–0

Job/Unit: O20041
/KAP1
Date: 10-04-12 15:13:49
Pages: 13
β-Linked Mono- and Divalent Mannosides
Click Chemistry
Three different β-linked divalent mannos-
ides, along with their monovalent counter-
parts, have been designed and chemically
synthesized by using click chemistry. The
synthesized saccharides were biologically
screened through a competitive inhibition
enzyme-linked immunosorbent assay to de-
termine the inhibition of specific human
IgG binding to low-molecular-weight hy-
drolyzed Candida albicans mannan.
C. Mukherjee, K. Ranta, J. Savolainen,
R. Leino* ........................................ 1–13
Synthesis and Immunological Screening of
β-Linked Mono- and Divalent Mannosides
Keywords: Medicinal chemistry / Carbo-
hydrates / Glycosylation / Click chemistry /
Biological activity
Eur. J. Org. Chem. 0000, 0–0
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