Synthesis of triazole-linked fluorescent saccharides and glycosyl amino esters

Article abstract of DOI:10.1055/s-0031-1290975

Synthesis of fluorescent carbohydrates and glycopeptides has been emerging as an attractive research field because of various biological roles of carbohydrates and glycoproteins. Fluorescent glycosides and C-glycosides were synthesized by copper(I)-catalyzed Huisgen reaction between 6-O-(2-azidoacetyl) glycopyranosides and alkyne-functionalized fluorophores. For the fluorescent disaccharides as well as glycosyl amino acids derivatives, glycosylation of azido-functionalized thioglycosides, followed by click reaction with corresponding fluorophores is preferred than the inversed procedure (click reaction before the glycosylation). Fluorophores like dansyl, NBD, or rhodamine have been successfully introduced on the sugar ring. All the newly synthesized fluorescent glycosides and glycoconjugates showed similar photophysical properties to the native fluorophores, with a higher quantum yield for the dansyl derivatives. Georg Thieme Verlag Stuttgart · New York.

Full text of DOI:10.1055/s-0031-1290975

PAPER
▌1647
paper
Synthesis of Triazole-Linked Fluorescent Saccharides and Glycosyl Amino
Esters
Synthesis of Triazole-Linked Fluorescent Saccharides
Hua Yi, Stéphane Maisonneuve, Juan Xie*
PPSM, ENS Cachan, CNRS, UMR 8531, 61 av. Président Wilson, 94235 Cachan cedex, France
Fax +33(1)47402454; E-mail: joanne.xie@ens-cachan.fr
Received: 25.01.2012; Accepted after revision: 24.03.2012
per(I)-catalyzed Huisgen 1,3-dipolar cycloaddition (Cu-
Abstract: Synthesis of fluorescent carbohydrates and glycopep-
AAC, also qualified as click reaction).4,8,15 Although the
tides has been emerging as an attractive research field because of
CuAAC reaction has been used to synthesize the fluores-
cent sugars, the reactivity of triazole-linked fluorescent
various biological roles of carbohydrates and glycoproteins. Fluo-
rescent glycosides and C-glycosides were synthesized by copper(I)-
catalyzed Huisgen reaction between 6-O-(2-azidoacetyl) glycopy- glycosyl donors have never been investigated. As a con-
ranosides and alkyne-functionalized fluorophores. For the fluores-
cent disaccharides as well as glycosyl amino acids derivatives,
glycosylation of azido-functionalized thioglycosides, followed by
we were interested in the synthesis of fluorescent sugars
click reaction with corresponding fluorophores is preferred than the
inversed procedure (click reaction before the glycosylation). Fluo-
rophores like dansyl, NBD, or rhodamine have been successfully in-
troduced on the sugar ring. All the newly synthesized fluorescent
glycosides and glycoconjugates showed similar photophysical
properties to the native fluorophores, with a higher quantum yield
for the dansyl derivatives.
tinuing program on the synthesis of carbohydrate conju-
gates and amino acid derivatives with click chemistry,16
and glycopeptides from azido-functionalized sugars as the
common starting materiel.
In order to functionalize the sugar ring with a fluorophore
and also realize the glycosylation reaction so as to obtain
fluorescent oligosaccharides as well as glycosyl amino
acids, phenyl thioglycoside has been chosen as the glyco-
syl donor due to its compatibility with click reaction con-
ditions. Since the 6-hydroxy-free thioglycosides are
readily accessible, we decided to introduce the azido func-
tion through an ester linkage. From azido-functionalized
thioglycosides, the target molecules are accessible either
through click reaction with fluorophores followed by gly-
cosylation with different glycosyl acceptors or through
glycosylation before introduction of different fluoro-
phores (Scheme 1). As fluorophores, dansyl, rhodamine,
and NBD (7-nitrobenz-2-oxa-1,3-diazole) chromophores
have been chosen because of their excellent photophysical
properties and fluorescence in the visible. Herein we re-
port the investigation of these two synthetic approaches
and photophysical properties of synthesized fluorescent
sugar derivatives.
Key words: fluorescence, saccharides, glycosyl amino acids, cop-
per(I), Huisgen cycloaddition reaction, glycosylation
Oligosaccharides and glycoconjugates play key roles in
various biological events, including signal transduction,
cell–cell communication, cancer metastasis, as well as
bacterial and viral infection.1 Development of chemical
tools is indispensable in order to understand and control
the biological functions of carbohydrates. Fluorescent la-
beling of biomolecules has been proven to be a powerful
method to investigate various biological phenomena at
molecular level because of the high sensitivity and effi-
ciency of fluorescence spectroscopy.2 Different aspects of
fluorescence output like steady-state intensity, Stokes
shift, life-time, steady-state and time-resolved anisotro-
pies, and resonance energy transfer could be used for
studying biological structures, dynamics, and functions,
and for visualizing intracellular processes or molecular in-
teractions.3 Consequently, synthesis of fluorescent carbo-
hydrates and glycoconjugates has attracted increasing
attention since the last decade. Fluorescent sugar deriva-
tives have been utilized as protein ligands,4 enzyme sub-
strates or inhibitors,5 molecular probes for the
investigation of cell membrane,6 glucose uptake,7 protein
glycosylation8 or for the direct observation of target cells.9
Fluorophores have been introduced into the sugar moiety
through amide/ester formation,4–7,9,10 reductive amina-
tion,11 Sonogashira coupling,12 aldol reaction,13 oxime-
based synthesis,14 as well as the recently developed cop-
Azido group was first introduced into the 6-position of
thioglycosides through esterification of phenyl β-D-gluco-
and galactopyranosides 1 and 2 with chloroacetyl chloride
followed by treatment with sodium azide. The desired az-
ido derivatives 3 and 4 were obtained in excellent yields
(Scheme 2). Click reaction of 3 or 4 with NBD alkyne 516h
in the presence of CuSO4/Na ascorbate afforded the NBD-
functionalized thioglycosides 6 and 7, with a better yield
for the gluco derivative. Similarly, click reaction with
dansyl alkyne 816h furnished the corresponding dansyl
sugars 9 and 10. We have also prepared the rhodamine al-
kyne 1117 by DCC-catalyzed coupling of rhodamine B
with propargylamine in 62% yield. Cycloaddition be-
tween 3 or 4 with rhodamine alkyne 11 led to 12 and 13.
With the fluorescent thioglycosides in hand, we have then
tried to realize the glycosylation reaction so as to investi-
gate the possibility of synthesis of fluorescent glycocon-
jugates. Glycosylation of 7 with diacetone α-D-galactose
using NIS in the presence of TfOH in MeCN18 led, how-
SYNTHESIS 2012, 44, 1647–1656
Advanced online publication: 09.05.2012
0
0
3
9
-
7
8
8
1
1
4
3
7
-
2
1
0
X
DOI: 10.1055/s-0031-1290975; Art ID: SS-2012-Z0080-OP
© Georg Thieme Verlag Stuttgart · New York

1648
H. Yi et al.
PAPER
N
N
N
O
O
N
O
a
b
b
N
N
O
SPh
O
O
N3
O
(OBn)3
O
O
O
R
SPh
O
a
N3
O
O
(OBn)3
(OBn)3
O
R
(OBn)3
H
Et2N
O
NEt2
N
NH
SO2
N
N
: fluorophore
O
N
NO2
NMe2
O
OH
O
CO2Bn
O
O
HO
ROH :
O
NHCbz
O
a: glycosylation with ROH
b: click reaction with
Scheme 1 Retrosynthesis of fluorescent saccharides and glycosyl amino acids derivatives
ever, to a mixture of products, whatever the reaction con-
ditions (quantity of catalyst, temperature, concentration,
etc.). Elimination of the NBD fluorophore from the sugar
unit has also been observed.
Table 1 Photophysical Properties of Fluorescent Saccharides
Solvent
λabs (nm) λem (nm)a Δ (cm–1) ΦF
8
9
DMSO–H2O
EtOH
328
330
341
340
341
338
342
338
466
451
460
454
314
314
315
315
569
532
496
527
495
527
498
529
551
528
566
531
466
427
472
469
12913
1506
0.20b
0.84b
0.30b
0.95b
0.38b
0.98b
0.60b
0.96b
0.01c
0.23c
0.04c
0.16c
0.09b
0.07b
0.06b
0.07b
To circumvent the difficulty, it was decided to realize first
the glycosylation reaction before introduction of the fluo-
rophore (Scheme 3). Glycosylation of 3 or 4 with diace-
tone α-D-galactose using NIS in the presence of TfOH in
MeCN at –40 °C afforded stereoselectively the corre-
sponding β-disaccharides 14 (α/β = 1:10) and 15 (only β)
in excellent yields. Click reaction of 14 or 15 with NBD
alkyne 5, dansyl alkyne 8, and rhodamine alkyne 11 took
place smoothly to afford the corresponding fluorescent di-
saccharides 16 to 21 in good yields.
DMSO–H2O
EtOH
9164
10436
9124
24
18
5
DMSO–H2O
EtOH
10611
9159
DMSO–H2O
EtOH
10682
3310
We have also tried to prepare fluorescent glycosyl amino
acid derivatives (Scheme 4). Glycosylation of thiogluco-
side 3 with CbzSerOBn by employing NIS/TfOH in
MeCN at –40 °C led, however, to compound 22 as a mix-
ture of anomers (α/β = 1:3). A better selectivity was ob-
tained with the galacto derivative 23 (α/β = 1:9).
Cycloaddition reaction of 22 or 23 with alkynes 8 and 11
afforded the corresponding fluorescent glycosyl amino
esters 24 to 27 in good yields.
DMSO–H2O
EtOH
3234
7
DMSO–H2O
EtOH
4071
3194
11
13
21
27
CH2Cl2
10387
8428
CH2Cl2
C-Allyl glycosides have proven to be a very useful syn-
thetic intermediates.19 Synthesis of fluorescent C-allyl
glycosides may contribute to the development of various
enzymatically stable glycomimetics. The azido-function-
alized α-C-allyl glucoside 29 was prepared either from the
α-C-allyl glucoside 2819a (chloroacetylation followed by
CH2Cl2
10560
10424
CH2Cl2
a Excitation was set equal to λabs
.
b Quantum yields (ΦF) were determined using as reference a solution
of quinine sulfate.
azidation) or from methyl α-D-glucoside 30 (chloroacety- c Quantum yields (ΦF) were determined using as reference a solution
of coumarin 153.
Synthesis 2012, 44, 1647–1656
© Georg Thieme Verlag Stuttgart · New York

PAPER
Synthesis of Triazole-Linked Fluorescent Saccharides
1649
lation, azidation then α-C-allylation with TMSAllyl– solvent polarity, with a blue shift for the dansyl alkyne 8
TMSOTf) (Scheme 5). Subsequent click reaction with 8 and NBD derivatives 5, 7, and a red shift for compounds
or 11 afforded the corresponding glucosides 31 and 32.
9, 18, and 24 in ethanol. All three tested dansyl glycocon-
jugates, which had quantitatively similar absorption and
emission spectra, displayed a red shift of about 10 nm in
absorption and a higher quantum yield as compared to the
native molecule 8. Only slight difference could be ob-
served in the absorption and emission spectra between
NBD derivatives 5 and 7. Due to the spirolactam struc-
ture, the rhodamine derivatives 11, 13, 21, and 27 scarcely
showed absorption or emission bands in the visible re-
gion. Further photophysical study of these rhodamine de-
rivatives is ongoing in our laboratory.
The photophysical properties of synthetic compounds
were then investigated in DMSO–water (2:8), ethanol, or
CH2Cl2. The fluorescence quantum yields were deter-
mined by using quinine sulfate for dansyl and rhodamine
derivatives, and coumarin 153 for NBD derivatives. As
shown in Table 1, spectroscopic properties of the newly
synthesized NBD 7 and dansyl 9, 18, and 24 glycoconju-
gates were similar to the native fluorophores 5 and 8: they
exhibited large Stokes shifts and higher quantum yields in
ethanol. Their fluorescent emissions were also affected by
O
R1
R1
N3
OH
O
a, b
R2
BnO
R2
BnO
O
O
SPh
SPh
OBn
OBn
1: R1 = H, R2 = OBn
3: R1 = H, R2 = OBn, 94%
2: R1 = OBn, R2 = H
4: R1 = OBn, R2 = H, 93%
N
HN
NH
O
N
N
R1
O
c
N
N
N
3 or 4
+
O
O
R2
BnO
O
N
SPh
NO2
5
NO2
N
OBn
6: R1 = H, R2 = OBn, 96%
7: R1 = OBn, R2 = H, 79%
H
N
H
N
SO2
O
N
SO2
R1
O
c
N
3 or 4 +
R2
BnO
O
SPh
NMe2
OBn
NMe2
9: R1 = H, R2 = OBn, 91%
10: R1 = OBn, R2 = H, 59%
8
+
NEt2
Et2N
Et2N
NEt2
O
O
Cl–
d
N
62%
CO2H
O
rhodamine B
11
Et2N
N
NEt2
O
O
N
N
c
R1
3 or 4 + 11
O
N
O
R2
BnO
O
SPh
OBn
12: R1 = H, R2 = OBn, 91%
13: R1 = OBn, R2 = H, 69%
OH
O
O
e
O
7 +
O
O
Scheme 2 Reagents and conditions: (a) chloroacetyl chloride, Et3N, CH2Cl2; (b) NaN3, NaI, acetone; (c) Na ascorbate, CuSO4, CH2Cl2, H2O;
(d) propargylamine, Et3N, DCC, HOBt, CH2Cl2; (e) NIS, TfOH, 4 Å MS, MeCN.
© Georg Thieme Verlag Stuttgart · New York
Synthesis 2012, 44, 1647–1656

1650
H. Yi et al.
PAPER
O
O
R1
R1
N3
O
N3
O
O
R2
CO2Bn
NHCbz
O
R2
O
OH
O
CO2Bn
NHCbz
O
BnO
BnO
HO
a
O
O
3 or 4 +
a
OBn
OBn
O
O
3 or 4 +
22: R1 = H, R2 = OBn, 78%
23: R1 = OBn, R2 = H, 75%
O
O
O
O
O
O
14: R1 = H, R2 = OBn, 96%
15: R1 = OBn, R2 = H, 99%
H
N
N
N
N
O
SO2
R1
O
N
NH
N
N
O
R2
BnO
b
O
R1
O
22 or 23 + 8
N
N
O
CO2Bn
R2
O
OBn
b
NMe2
NEt2
O
BnO
CbzHN
14 or 15 + 5
24: R1 = H, R2 = OBn, 93%
25: R1 = OBn, R2 = H, 76%
OBn
O
NO2
O
O
O
Et2N
O
O
16: R1 = H, R2 = OBn, 61%
17: R1 = OBn, R2 = H, 74%
N
N
O
N
R1
O
H
N
O
N
N
SO2
R2
BnO
O
N
N
O
R1
O
O
b
CO2Bn
22 or 23 + 11
OBn
R2
BnO
b
O
CbzHN
14 or 15 + 8
O
26: R1 = H, R2 = OBn, 98%
27: R1 = OBn, R2 = H, 91%
OBn
O
NMe2
O
Scheme 4 Reagents and conditions: (a) NIS, TfOH, 4 Å MS, MeCN,
O
O
–40 °C; (b) Na ascorbate, CuSO4, CH2Cl2, H2O.
O
18: R1 = H, R2 = OBn, 98%
19: R1 = OBn, R2 = H, 76%
aluminum percolated plates of Silica Gel 60F-254 with detection by
UV and by spraying with 10% H2SO4 in EtOH solution and heating
for about 20 sec at 400 °C. Petroleum ether (PE) used refers to the
Et2N
N
NEt2
O
1
fraction boiling in the 40–65 °C range. H and 13C NMR spectra
N
N
O
N
R1
O
were recorded on a JEOL ECS 400 MHz spectrometer. The spectra
were taken with TMS as a proton standard reference (0 ppm). For
13C spectra, central peak of the residual solvent multiplets were used
as a chemical shift reference (CDCl3: 77.00 ppm). ESI-HRMS mass
spectra were recorded on a Bruker micrOTOF-Q II spectrometer.
UV/vis absorption spectra were recorded on a Uvikon-940 KON-
TRON spectrophotometer. The fluorescence quantum yields (ΦF)
were determined by the standard method using a solution of 1.0 ×
10–5 M quinine sulfate in 0.5 N H2SO4 or coumarin 153 in EtOH as
reference. Only diluted and degassed solutions with an absorbance
below 0.1 at the excitation wavelength λex were used.
O
R2
O
O
BnO
OBn
O
b
O
14 or 15 + 11
O
O
O
20: R1 = H, R2 = OBn, 99%
21: R1 = OBn, R2 = H, 89%
Scheme 3 Reagents and conditions: (a) NIS, TfOH, 4 Å MS, MeCN,
–40 °C; (b) Na ascorbate, CuSO4, CH2Cl2, H2O.
6-O-Azidoacetyl Glycopyranosides; General Procedure 1 (GP
1)
To a solution of partially protected glycopyranoside (0.3 mmol) and
Et3N (0.63 mmol) in CH2Cl2 (10 mL) at 0 °C, was added chloroace-
tyl chloride (0.6 mmol). The mixture was stirred for 2 h at 0 °C and
then the temperature was gradually raised to r.t. and stirred for 12 h.
After evaporation, the residue was dissolved in EtOAc (20 mL), and
the EtOAc layer was washed with sat. aq NaHCO3 (3 × 10 mL) and
brine (3 × 10 mL). The organic layer was dried (MgSO4) and con-
centrated under vacuum to give a crude product, which was used
without purification in the next step. To a solution of crude 6-O-
chloroacetyl glycopyranoside in acetone (10 mL), was added NaN3
(1.5 mmol) and KI (10 mg). After stirring for 24 h at r.t., the mixture
was evaporated. The residue was dissolved in EtOAc (20 mL), and
the EtOAc layer was washed with sat. aq NaHCO3 (3 × 10 mL) and
brine (3 × 10 mL). The organic layer was dried (MgSO4) and con-
centrated under vacuum to give a crude product, which was purified
by column chromatography.
In summary, the copper(I)-catalyzed Huisgen 1,3-dipolar
cycloaddition has been successfully used for the synthesis
of fluorescent glycosides and C-glycosides by reaction of
6-O-(2-azidoacetyl) glycopyranosides with dansyl, NBD,
or rhodamine alkynes. For the synthesis of fluorescent di-
saccharides as well as glycosyl amino esters, it is pre-
ferred to realize the glycosylation of 6-O-(2-azidoacetyl)
thioglycosides first before the introduction of fluorophore
with click reaction. All the newly synthesized fluorescent
glycosides and glycoconjugates showed similar photo-
physical properties to the native fluorophores, with a
higher quantum yield for dansyl derivatives.
Melting points (Mp) were obtained without correction using a
Kofler melting point apparatus. Column chromatography was per-
formed on silica gel 40–63 μm. Analytical TLC was performed on
Glycosylation; General Procedure 2 (GP 2)
A mixture of thioglycoside (0.5 mmol), glycosyl acceptor (0.55
mmol), NIS (0.6 mmol), and powdered 4 Å molecular sieves (1 g)
Synthesis 2012, 44, 1647–1656
© Georg Thieme Verlag Stuttgart · New York

PAPER
Synthesis of Triazole-Linked Fluorescent Saccharides
1651
O
N3
O
OH
OH
a, b
a, b, c
O
O
O
BnO
BnO
BnO
BnO
BnO
BnO
70%
49%
BnO
BnO
OMe
BnO
30
28
29
H
N
N
N
N
O
SO2
O
d
O
BnO
BnO
29 + 8
93%
BnO
NMe2
31
Et2N
NEt2
O
N
N
N
O
N
O
O
d
O
29 + 11
BnO
BnO
99%
BnO
32
Scheme 5 Reagents and conditions: (a) chloroacetyl chloride, Et3N, CH2Cl2; (b) NaN3, NaI, acetone; (c) TMSallyl, TMSOTf, MeCN; (d) Na
ascorbate, CuSO4, CH2Cl2, H2O.
in MeCN (10 mL) was stirred for 30 min at –40 °C before the addi-
tion of TfOH (0.015 mmol). The mixture was stirred at –40 °C for
2 h and the temperature was then gradually raised to r.t. After the
completion of the reaction, Na2SO3 (1 g), NaHCO3 (1 g), and a few
drops of H2O were added to the reaction mixture. The mixture was
stirred for 5 min, then diluted with CH2Cl2 (25 mL), filtered through
Celite, and the CH2Cl2 layer was washed with sat. aq NaHCO3 (3 ×
10 mL) and brine (3 × 10 mL). The organic layer was dried
(MgSO4) and concentrated in vacuum to give a crude product,
which was purified by chromatography.
HRMS (ESI): m/z calcd for C35H35N3O6S + Na [M + Na+]:
648.2144; found: 648.2127.
Phenyl 6-O-(2-Azido)acetyl-2,3,4-tri-O-benzyl-1-thio-β-D-ga-
lactopyranoside (4)
Phenyl 2,3,4-tri-O-benzyl-1-thio-β-galactopyranoside (2; 1.012 g,
1.867 mmol) was treated successively with ClCH2COCl and NaN3
according to GP 1. Purification by column chromatography
(EtOAc–PE, 1:9) gave 1.060 g (93%) of compound 4 as a white sol-
id; mp 84 °C; [α]D20 –21.3 (c 0.5, CHCl3).
1H NMR (400 MHz, CDCl3): δ = 3.58–3.62 (m, 2 H, H-3,5), 3.66
(d, J = 17.0 Hz, 1 H), 3.74 (d, J = 17.0 Hz, 1 H), 3.83 (d, J = 1.8 Hz,
1 H, H-4), 3.95 (t, J = 9.6 Hz, 1 H, H-2), 4.13 (dd, J = 5.5, 11.4 Hz,
1 H, H-6), 4.36 (dd, J = 6.9, 11.0 Hz, 1 H, H-6′), 4.62 (d, J = 9.6 Hz,
1 H, H-1), 4.76 (d, J = 11.9 Hz, 1 H), 4.74–4.84 (m, 4 H), 4.99 (d,
J = 11.9 Hz, 1 H), 7.23–7.40 (m, 18 H), 7.53–7.59 (m, 2 H).
13C NMR (100 MHz, CDCl3): δ = 50.2, 64.8, 73.0, 73.4, 74.3, 75.8,
75.9, 77.3, 84.2, 88.0, 127.8, 128.0, 128.4, 128.5, 128.6, 128.7,
128.9, 132.0, 133.9, 138.1, 138.2, 168.0.
Click Reaction; General Procedure 3 (GP 3)
To a solution of alkyne (0.05 mmol) and azide (0.05 mmol) in
CH2Cl2 (5 mL) and H2O (5 mL), were added CuSO4·5H2O (0.01
mmol) and Na ascorbate (0.01 mmol) successively. The reaction
mixture was vigorously stirred at r.t. for 12 to 20 h. CH2Cl2 (10 mL)
was then added. After separation of the organic layer, the aqueous
layer was extracted with CH2Cl2 (3 × 10 mL). The combined organ-
ic layers were washed with brine (2 × 20 mL), dried (MgSO4), evap-
orated, and the residue purified by column chromatography.
HRMS (ESI): m/z calcd for C35H35N3O6S + Na [M + Na+]:
648.2144; found: 648.2126.
Phenyl 6-O-(2-Azido)acetyl-2,3,4-tri-O-benzyl-1-thio-β-D-glu-
copyranoside (3)
Phenyl 2,3,4-tri-O-benzyl-1-thio-β-D-glucopyranoside (1; 1.112 g,
2.049 mmol) was treated successively with ClCH2COCl and NaN3
according to GP 1. Purification by column chromatography
(EtOAc–PE, 1:9) gave 1.210 g (94%) of compound 3 as a white sol-
id; mp 65 °C; [α]D20 +26.8 (c 0.5, CHCl3).
1H NMR (400 MHz, CDCl3): δ = 3.47–3.57 (m, 3 H, H-2,4,5), 3.72
(t, J = 8.2 Hz, 1 H, H-3), 3.73 (d, J = 17.0 Hz, 1 H), 3.79 (d, J = 17.0
Hz, 1 H), 4.27 (dd, J = 5.3, 11.9 Hz, 1 H, H-6), 4.43 (dd, J = 1.8,
11.9 Hz, 1 H, H-6′), 4.59 (d, J = 11.4 Hz, 1 H), 4.65 (d, J = 9.6 Hz,
1 H, H-1), 4.74 (d, J = 10.1 Hz, 1 H), 4.85 (d, J = 11.0 Hz, 1 H), 4.86
(d, J = 11.0 Hz, 1 H), 4.91 (d, J = 10.1 Hz, 1 H), 4.94 (d, J = 11.0
Hz, 1 H), 7.25–7.39 (m, 18 H), 7.51–7.53 (m, 2 H).
Phenyl 2,3,4-Tri-O-benzyl-6-O-{4-[(7-nitrobenzo[1,2,5]oxadia-
zol-4-ylamino)methyl]-1H-1,2,3-triazol-1-yl}acetyl-1-thio-β-D-
glucopyranoside (6)
From 7-nitro-N-(prop-2-ynyl)benzo[c][1,2,5]oxadiazol-4-amine
(5; 30 mg, 0.13 mmol) and azidoacetyl thioglucoside 3 (34 mg,
0.054 mmol), click reaction according to GP 3 and purification by
column chromatography (EtOAc–PE, 2:8) led to 6 as an orange sol-
id (44 mg, 96%); mp 85 °C; [α]D20 +16.5 (c 0.25, CHCl3).
1H NMR (400 MHz, CDCl3): δ = 3.42 (t, J = 10.1 Hz, 1 H, H-2),
3.46 (t, J = 9.6 Hz, 1 H, H-4), 3.50–3.53 (m, 1 H, H-5), 3.72 (t, J =
8.7 Hz, 1 H, H-3), 4.23 (dd, J = 5.5, 11.4 Hz, 1 H, H-6), 4.45 (dd,
J = 1.8, 11.5 Hz, 1 H, H-6′), 4.53 (d, J = 11.0 Hz, 1 H, H-1), 4.65 (d,
J = 9.6 Hz, 1 H), 4.73–4.95 (m, 7 H), 5.03 (d, J = 17.4 Hz, 1 H), 5.12
(d, J = 17.4 Hz, 1 H), 6.30 (d, J = 8.2 Hz, 1 H, ArH), 6.75 (t, J = 5.5
Hz, 1 H, NH), 7.23–7.48 (m, 20 H), 7.64 (s, 1 H), 8.44 (d, J = 8.2
Hz, 1 H).
13C NMR (100 MHz, CDCl3): δ = 50.2, 64.4, 75.1, 75.6, 76.0, 76.6,
77.1, 86.8, 87.6, 127.8, 127.9, 128.3, 128.6, 128.7, 129.0, 132.3,
137.6, 137.9, 138.2, 168.0.
© Georg Thieme Verlag Stuttgart · New York
Synthesis 2012, 44, 1647–1656

1652
H. Yi et al.
PAPER
13C NMR (100 MHz, CDCl3): δ = 39.5, 50.9, 64.9, 75.1, 75.6, 75.9,
76.9, 80.9, 86.7, 87.6, 99.7, 125.5, 127.9, 128.3, 128.3, 128.6,
128.7, 129.0, 132.1, 133.3, 136.1, 138.0, 142.7, 144.4, 165.7.
HRMS (ESI): m/z calcd for C44H41N7O9S + Na [M + Na+]:
866.2584; found: 866.2562.
13C NMR (100 MHz, CDCl3): δ = 38.8, 45.5, 50.6, 60.5, 64.1, 73.1,
73.3, 74.3, 75.6, 75.8, 77.3, 84.0, 87.7, 115.4, 118.8, 123.3, 124.0,
127.5, 127.7, 127.9, 128.3, 128.4, 128.5, 128.6, 128.7, 129.0, 129.7,
131.8, 133.9, 138.1, 138.3, 144.6, 152.1, 165.9.
HRMS (ESI): m/z calcd for C50H51N5O8S2 + Na [M + Na+]:
936.3077; found: 936.3054.
Phenyl 2,3,4-Tri-O-benzyl-6-O-{4-[(7-nitrobenzo[1,2,5]oxadia-
zol-4-ylamino)methyl]-1H-1,2,3-triazol-1-yl}acetyl-1-thio-β-D-
galactopyranoside (7)
3′,6′-Bis(diethylamino)-2-(prop-2-ynyl)spiro[isoindoline-1,9′-
xanthen]-3-one (11)
From 5 (36 mg, 0.165 mmol) and azidoacetyl thiogalactoside 4 (102
mg, 0.163 mmol), click reaction according to GP 3 and purification
by column chromatography (EtOAc–PE, 5:5 to 7:3) led to 7 as or-
ange pellets (110 mg, 79%); mp 100 °C (dec.); [α]D20 –5.5 (c 0.33,
CHCl3).
1H NMR (400 MHz, CDCl3): δ = 3.61–3.65 (m, 2 H, H-3,5), 3.82
(d, J = 1.8 Hz, 1 H, H-4), 3.95 (t, J = 9.6 Hz, 1 H, H-2), 4.09 (dd,
J = 4.6, 11.4 Hz, 1 H, H-6), 4.66 (dd, J = 7.8, 11.4 Hz, 1 H, H-6′),
4.59 (d, J = 11.9 Hz, 1 H), 4.63 (d, J = 9.6 Hz, 1 H, H-1), 4.73–4.81
(m, 6 H), 5.00 (d, J = 11.4 Hz, 1 H), 5.03 (d, J = 17.4 Hz, 1 H), 5.09
(d, J = 17.4 Hz, 1 H), 6.30 (d, J = 8.7 Hz, 1 H, ArH), 7.03 (m, 1 H,
NH), 7.20–7.40 (m, 18 H), 7.51–7.53 (m, 2 H), 7.72 (s, 1 H), 8.41
(d, J = 9.7 Hz, 1 H, ArH).
13C NMR (100 MHz, CDCl3): δ = 39.4, 50.8, 60.5, 65.5, 73.3, 73.4,
74.3, 75.7, 75.8, 77.4, 84.1, 87.7, 99.8, 123.9, 124.8, 127.5, 127.8,
128.0, 128.3, 128.4, 128.5, 128.7, 129.0, 133.9, 136.3, 138.0, 138.2,
142.8, 143.3, 143.9, 144.4, 165.9, 171.3.
HRMS (ESI): m/z calcd for C44H41N7O9S + Na [M + Na+]:
866.2584; found: 866.2581.
To a solution of rhodamine B (196 mg, 0.4 mmol) in CH2Cl2 (25
mL) at 0 °C were added Et3N (0.2 mL, 1.4 mmol), propargylamine
(102 μL, 0.402 mmol), DCC (160 mg, 0.8 mmol), and HOBt (80
mg, 0.6 mmol). After stirring 18 h at r.t., the reaction mixture was
filtered and the filtrate evaporated. The residue was dissolved in
EtOAc (20 mL) and the solution was washed successively with H2O
(10 mL) and brine (10 mL), dried (MgSO4), evaporated, and puri-
fied by column chromatography (EtOAc–PE, 1:9 to 1:4) to give 120
mg of 11 (62%) as a pink solid; mp 163 °C.
1H NMR (400 MHz, CDCl3): δ = 1.15 (t, J = 6.9 Hz, 12 H, 4 × CH3),
1.75 (t, J = 2.8 Hz, 1 H, CH≡), 3.24 (q, J = 6.9 Hz, 8 H, 4 × CH2),
3.93 (d, J = 2.4 Hz, 2 H, CH2), 6.26 (dd, J = 2.7, 9.2 Hz, 2 H, ArH),
6.37 (d, J = 2.3 Hz, 2 H, ArH), 6.45 (d, J = 8.7 Hz, 2 H, ArH), 7.07–
7.11 (m, 1 H, ArH), 7.40–7.44 (m, 2 H, ArH), 7.89–7.93 (m, 1 H,
ArH).
13C NMR (100 MHz, CDCl3): δ = 12.7, 28.7, 44.5, 64.9, 70.1, 77.5,
97.9, 105.2, 108.1, 123.2, 123.9, 128.1, 129.3, 130.6, 132.8, 149.1,
153.6, 153.9, 167.5.
HRMS (ESI): m/z calcd for C31H34N3O2 [M + H+]: 480.2651; found:
480.2633.
Phenyl 2,3,4-Tri-O-benzyl-6-O-{4-[(5-dimethylaminonaphtha-
lene-1-sulfonylamino)methyl]-1H-1,2,3-triazol-1-yl}acetyl-1-
thio-β-D-glucopyranoside (9)
Phenyl 2,3,4-Tri-O-benzyl-6-O-[2-(4-{[3′,6′-bis(diethylamino)-
3-oxospiro(isoindoline-1,9′-xanthene)-2-yl]methyl}-1H-1,2,3-
triazol-1-yl)]acetyl-1-thio-β-D-glucopyranoside (12)
From 11 (12 mg, 0.025 mmol) and azidoacetyl thioglucoside 3 (13
mg, 0.021 mmol), click reaction according to GP 3 and purification
by column chromatography (EtOAc–PE, 1:9 to 1:4) led to 12 as a
From 5-(dimethylamino)-N-(prop-2-ynyl)naphthalene-1-sulfon-
amide (8; 28 mg, 0.097 mmol) and azidoacetyl thioglucoside 3 (38
mg, 0.06 mmol), click reaction according to GP 3 and purification
by column chromatography (EtOAc–PE, 7:3) led to 9 as a
20
20
red/brown solid (51 mg, 91%); mp 64 °C; [α]D +34.4 (c 0.5,
pink solid (21 mg, 91%); mp 102–104 °C; [α]D +24.5 (c 0.5,
CHCl3).
CHCl3).
1H NMR (400 MHz, CDCl3): δ = 2.89 (s, 6 H, 2 × CH3), 3.39 (t, J =
8.9 Hz, 1 H, H-4), 3.48–3.53 (m, 2 H, H-2,5), 3.72 (t, J = 8.9 Hz, 1
H, H-3), 4.17–4.21 (m, 3 H, H-6, NCH2), 4.43 (dd, J = 2.3, 11.9 Hz,
1 H, H-6′), 4.47 (d, J = 11.0 Hz, 1 H), 4.65 (d, J = 10.1 Hz, 1 H, H-
1), 4.76–4.99 (m, 7 H), 5.38 (t, J = 6.0 Hz, 1 H, NH), 7.25–7.51 (m,
24 H), 8.20 (dd, J = 0.9, 7.4 Hz, 1 H), 8.25 (d, J = 8.7 Hz, 1 H), 8.52
(d, J = 8.3 Hz, 1 H).
13C NMR (100 MHz, CDCl3): δ = 38.9, 45.5, 50.6, 64.8, 75.0, 75.6,
75.9, 76.4, 77.0, 80.9, 86.7, 87.5, 115.4, 118.7, 123.2, 123.7, 127.8,
127.9, 128.0, 128.2, 128.3, 128.6, 128.7, 129.1, 129.7, 129.9, 130.7,
132.3, 133.2, 134.5, 137.5, 137.9, 138.1, 144.6, 152.1, 165.7.
1H NMR (400 MHz, CDCl3): δ = 1.14 (t, J = 6.8 Hz, 12 H, 4 × CH3),
3.28 (q, J = 6.8 Hz, 8 H, 4 × CH2), 3.55 (t, J = 9.2 Hz, 1 H), 3.46–
3.51 (m, 2 H), 3.71 (t, J = 8.7 Hz, 1 H), 4.17 (dd, J = 6.0, 11.9 Hz,
1 H, H-6), 4.39 (dd, J = 2.3, 11.9 Hz, 1 H, H-6′), 4.49 (s, 2 H,
NCH2), 4.51 (d, J = 11.0 Hz, 1 H, CHPh), 4.65 (d, J = 9.6 Hz, 1 H,
H-1), 4.73–4.85 (m, 5 H), 4.91–4.94 (m, 2 H, CH2), 6.13 (dd, J =
2.7, 8.7 Hz, 2 H), 6.29 (d, J = 8.7 Hz, 2 H, ArH), 6.34 (d, J = 2.8 Hz,
2 H, ArH), 7.09–7.51 (m, 24 H, ArH), 7.91–7.93 (m, 1 H, CH=).
13C NMR (100 MHz, CDCl3): δ = 12.7, 35.3, 44.4, 50.2, 64.7, 65.1,
75.0, 75.6, 75.9, 76.5, 77.3, 80.9, 86.7, 87.6, 97.9, 105.3, 108.0,
123.0, 123.9, 124.0, 127.9, 128.2, 128.3, 128.5, 128.6, 128.7, 128.9,
129.1, 131.0, 132.2, 132.6, 133.4, 137.5, 137.9, 138.2, 145.0, 148.8,
153.5, 153.7, 165.9, 168.0.
HRMS (ESI): m/z calcd for C50H51N5O8S2 + Na [M + Na+]:
936.3077; found: 936.3075.
HRMS (ESI): m/z calcd for C66H68N6O8S + Na [M + Na+]:
1127.4717; found: 1127.4649.
Phenyl 2,3,4-Tri-O-benzyl-6-O-{4-[(5-dimethylaminonaphtha-
lene-1-sulfonylamino)methyl]-1H-1,2,3-triazol-1-yl}acetyl-1-
thio-β-D-galactopyranoside (10)
Phenyl 2,3,4-Tri-O-benzyl-6-O-[2-(4-{[3′,6′-bis(diethylamino)-
3-oxospiro(isoindoline-1,9′-xanthene)-2-yl]methyl}-1H-1,2,3-
triazol-1-yl)]acetyl-1-thio-β-D-galactopyranoside (13)
From 8 (32 mg, 0.111 mmol) and azidoacetyl thiogalactoside 4 (22
mg, 0.054 mmol), click reaction according to GP 3 and purification
by column chromatography (EtOAc–PE, 5:5) led to 10 (29 mg,
From 11 (20 mg, 0.042 mmol) and azidoacetyl thiogalactoside 4 (31
mg, 0.050 mmol), click reaction according to GP 3 and purification
by column chromatography (EtOAc–PE, 1:9 to 1:4) led to 13 as a
20
59%) as a brown solid; mp 72–74 °C (dec.); [α]D –12.5 (c 0.5,
CHCl3).
20
1H NMR (400 MHz, CDCl3): δ = 2.89 (s, 6 H, 2 × CH3), 3.57–3.61
(m, 2 H, H-3,5), 3.80 (d, J = 1.8 Hz, 1 H, H-4), 3.93 (t, J = 9.6 Hz,
1 H, H-2), 4.08 (dd, J = 5.5, 11.5 Hz, 1 H, H-6), 4.16 (d, J = 6.4 Hz,
2 H, NCH2), 4.38 (dd, J = 7.4, 11.0 Hz, 1 H, H-6′), 4.58 (d, J = 11.4
Hz, 1 H), 4.62 (d, J = 9.6 Hz, 1 H, H-1), 4.73–4.81 (m, 4 H), 4.89
(s, 2 H, CH2), 4.98 (d, J = 11.9 Hz, 1 H), 5.68 (t, J = 6.0 Hz, 1 H,
NH), 7.12–7.56 (m, 24 H), 8.21–8.25 (m, 2 H), 8.51–8.53 (m, 1 H).
pink solid (32 mg, 69%); mp 103 °C (dec.); [α]D +3.0 (c 0.5,
CHCl3).
1H NMR (400 MHz, CDCl3): δ = 1.12 (t, J = 6.9 Hz, 12 H, 4 × CH3),
3.29 (q, J = 6.9 Hz, 8 H, 4 × CH2), 3.55 (t, J = 6.0 Hz, 1 H, H-5),
3.60 (dd, J = 2.7, 9.2 Hz, 1 H, H-3), 3.77 (m, 1 H, H-4), 3.93 (t, J =
9.6 Hz, 1 H, H-2), 4.06 (dd, J = 5.5, 11.4 Hz, 1 H, H-6), 4.34 (dd,
J = 6.7, 11.0 Hz, 1 H, H-6′), 4.46 (s, 2 H, NCH2), 4.57 (d, J = 11.9
Synthesis 2012, 44, 1647–1656
© Georg Thieme Verlag Stuttgart · New York

PAPER
Synthesis of Triazole-Linked Fluorescent Saccharides
1653
Hz, 1 H, CHPh), 4.62 (d, J = 9.6 Hz, 1 H, H-1), 4.71– 4.82 (m, 6 H,
NCH2, 2 × OCH2), 4.98 (d, J = 11.4 Hz, 1 H, CHPh), 6.13–6.16 (m,
2 H, ArH), 6.28–6.34 (m, 4 H, ArH), 7.07–7.54 (m, 24 H, ArH),
7.91–7.94 (m, 1 H, CH=).
13C NMR (100 MHz, CDCl3): δ = 12.7, 35.4, 44.5, 50.2, 64.9, 65.2,
73.1, 73.3, 74.3, 75.6, 75.8, 77.3, 84.1, 87.7, 97.9, 105.2, 108.1,
123.0, 123.9, 124.0, 127.8, 127.9, 128.2, 128.4, 128.5, 128.6, 128.9,
130.9, 131.8, 132.7, 133.8, 138.0, 138.2, 145.0, 148.8, 153.5, 153.6,
165.8, 168.0.
3.66 (t, J = 9.0 Hz, 1 H), 3.71 (dd, J = 8.5, 10.1 Hz, 1 H), 4.05–4.09
(m, 2 H), 4.20–4.24 (m, 2 H), 4.32 (dd, J = 2.3, 7.4 Hz, 1 H, H-2),
4.43–4.50 (m, 3 H), 4.58 (dd, J = 2.3, 7.8 Hz, 1 H, CH), 4.71 (d, J =
11.4 Hz, 1 H), 4.76 (d, J = 11.0 Hz, 1 H), 4.78–4.79 (m, 2 H), 4.81
(d, J = 11.4 Hz, 1 H), 4.97 (d, J = 11.0 Hz, 1 H), 5.03 (d, J = 11.0
Hz, 1 H), 5.08 (d, J = 17.4 Hz, 2 H, 2 × CH), 5.16 (d, J = 17.4 Hz,
2 H, 2 × CH), 5.56 (d, J = 5.0 Hz, 1 H, H-1), 6.31 (d, J = 8.7 Hz, 1
H), 7.18–7.41 (m, 15 H), 7.76 (s, 1 H), 8.39 (d, J = 8.2 Hz, 1 H).
13C NMR (100 MHz, CDCl3): δ = 24.5, 25.0, 26.08, 26.13, 39.4,
50.9, 64.9, 67.2, 69.8, 70.5, 70.8, 71.4, 72.4, 74.4, 74.8, 75.8, 76.7,
77.4, 81.4, 84.5, 96.4, 99.8, 104.3, 108.7, 109.5, 123.9, 124.6,
127.6, 127.6, 127.8, 128.0, 128.3, 128.5, 128.6, 128.8, 136.4, 137.7,
138.4, 138.5, 142.9, 143.4, 143.9, 144.3, 166.0.
HRMS (ESI): m/z calcd for C66H68N6O8S + Na [M + Na+]:
1127.4717; found: 1127.4714.
6-O-(2-Azido)acetyl-2,3,4-tri-O-benzyl-β-D-glucopyranosyl-
(1→6)-1,2:3,4-di-O-isopropylidene-α-D-galactopyranose (14)
Compound 3 (375 mg, 0.6 mmol) was treated with diacetone α-D-
galactose (178 mg, 0.68 mmol) according to GP 2. Purification by
chromatography (EtOAc–PE, 1:4) gave 446 mg (96%) of 14 (β/α =
10:1) as a colorless oil; [α]D20 –10.5 (c 1, CHCl3).
1H NMR (400 MHz, CDCl3): δ = 1.31, 1.32, 1.45, 1.49 (4 s, 12 H,
4 × CH3), 3.48 (t, J = 8.5 Hz, 1 H, H-2′), 3.49–3.50 (m, 2 H), 3.65–
3.75 (m, 2 H), 3.73 (d, J = 17.4 Hz, 1 H), 3.83 (d, J = 17.4 Hz, 1 H),
4.07–4.12 (m, 2 H), 4.22 (dd, J = 1.4, 8.2 Hz, 1 H), 4.30 (dd, J = 4.6,
8.7 Hz, 1 H), 4.32 (dd, J = 2.3, 5.0 Hz, 1 H), 4.41–4.44 (m, 1 H),
4.48 (d, J = 8.0 Hz, 1 H, H-1′), 4.56 (d, J = 11.4 Hz, 1 H), 4.60 (dd,
J = 2.2, 7.8 Hz, 1 H), 4.72 (d, J = 11.4 Hz, 1 H), 4.79 (d, J = 10.5
Hz, 1 H), 4.86 (d, J = 11.0 Hz, 1 H), 5.00 (d, J = 11.0 Hz, 1 H), 5.06
(d, J = 11.4 Hz, 1 H), 5.57 (d, J = 5.0 Hz, 1 H, H-1), 7.24–7.44 (m,
15 H).
13C NMR (100 MHz, CDCl3): δ = 24.5, 25.1, 26.1, 50.1, 64.3, 67.4,
70.1, 70.5, 70.9, 71.5, 72.6, 74.5, 74.9, 75.8, 76.8, 81.5, 84.6, 96.5,
104.5, 108.7, 109.5, 127.7, 127.8, 128.0, 128.2, 128.4, 128.5, 128.6,
128.7, 137.8, 138.5, 138.6, 168.6.
HRMS (ESI): m/z calcd for C41H49N3O12 + Na [M + Na+]: 798.3214;
found: 798.3203.
HRMS (ESI): m/z calcd for C50H56N7O15 [M + H+]: 994.3834;
found: 994.3822.
2,3,4-Tri-O-benzyl-6-O-{4-[(7-nitrobenzo[1,2,5]oxadiazol-4-yl-
amino)methyl]-1H-1,2,3-triazol-1-yl}acetyl-β-D-galactopyrano-
syl-(1→6)-1,2:3,4-di-O-isopropylidene-α-D-galactopyranose (17)
From NBD alkyne 5 (27 mg, 0.126 mmol) and azidoacetyl disac-
charide 15 (97 mg, 0.126 mmol), click reaction according to GP 3
and purification by column chromatography (EtOAc–PE, 1:4 to
3:7) led to 17 as an orange powder (92 mg, 74%); mp 66 °C (dec.);
[α]D20 +6.7 (c 0.5, CHCl3).
1H NMR (400 MHz, CDCl3): δ = 1.29 (s, 3 H, CH3), 1.30 (s, 3 H,
CH3), 1.42 (s, 3 H, CH3), 1.47 (s, 3 H, CH3), 3.50–3.53 (m, 2 H, H-
3′, H-4), 3.67 (dd, J = 6.9, 10.1 Hz, 1 H, H-6′), 3.72 (m, 1 H, H-5),
3.83 (dd, J = 7.8, 9.6 Hz, 1 H, H-2′), 4.03–4.12 (m, 3 H, H-3, H-5′,
H-6), 4.21 (dd, J = 1.6, 8.0 Hz, 1 H, CH), 4.31 (m, 2 H, H-2, H-4′),
4.40 (d, J = 7.8 Hz, 1 H, H-1′), 4.57 (dd, J = 2.3, 7.8 Hz, 1 H, CH),
4.60 (d, J = 12.7 Hz, 1 H, CH), 4.72 (d, J = 11.0 Hz, 1 H, CH), 4.74
(d, J = 11.9 Hz, 1 H, CH), 4.80 (d, J = 5.5 Hz, 2 H, CH2NH), 4.85
(d, J = 11.7 Hz, 1 H, CH), 4.94 (d, J = 11.9 Hz, 1 H, CH), 5.00–5.11
(m, 3 H), 5.55 (d, J = 5.0 Hz, 1 H, H-1), 6.31 (d, J = 8.7 Hz, 1 H),
7.13 (t, J = 5.5 Hz, 1 H, NH), 7.24–7.43 (m, 15 H), 7.76 (s, 1 H),
8.40 (d, J = 8.2 Hz, 1 H).
6-O-(2-Azido)acetyl-2,3,4-tri-O-benzyl-β-D-galactopyranosyl-
(1→6)-1,2:3,4-di-O-isopropylidene-α-D-galactopyranose (15)
Compound 4 (400 mg, 0.639 mmol) was treated with diacetone α-
D-galactose (189 mg, 0.726 mmol) according to GP 2. Purification
by chromatography (EtOAc–PE, 1:9–1:4) gave 490 mg (99%) of 15
as a white solid; mp 80 °C; [α]D20 –58.4 (c 0.5, CHCl3).
13C NMR (100 MHz, CDCl3): δ = 24.5, 25.1, 26.1, 39.4, 50.8, 65.2,
67.3, 69.5, 70.6, 70.8, 71.4, 71.8, 72.8, 73.7, 74.2, 74.9, 77.3, 78.9,
81.8, 96.4, 99.8, 104.5, 108.8, 109.5, 123.9, 124.7, 127.6, 127.7,
127.8, 128.0, 128.3, 128.5, 128.58, 128.55, 128.8, 136.4, 138.1,
138.4, 138.8, 142.8, 143.3, 143.9, 144.3, 165.9.
HRMS (ESI): m/z calcd for C50H56N7O15 [M + H+]: 994.3834;
found: 994.3816.
1H NMR (400 MHz, CDCl3): δ = 1.30, 1.31, 1.44, 1.48 (4 s, 12 H,
4 × CH3), 3.51–3.53 (m, 2 H, H-3, 3′), 3.62–3.74 (m, 4 H), 3.85 (dd,
J = 7.8, 9.6 Hz, 1 H, H-2′), 4.06–4.13 (m, 3 H), 4.21 (dd, J = 1.8, 7.8
Hz, 1 H), 4.26 (dd, J = 6.4, 11.0 Hz, 1 H), 4.30 (dd, J = 2.3, 5.0 Hz,
1 H, H-2), 4.41 (d, J = 7.4 Hz, 1 H, H-1′), 4.58 (dd, J = 2.3, 7.8 Hz,
1 H), 4.65 (d, J = 11.9 Hz, 1 H), 4.73 (d, J = 11.0 Hz, 1 H), 4.74 (d,
J = 11.9 Hz, 1 H), 4.85 (d, J = 11.9 Hz, 1 H), 4.94 (d, J = 11.9 Hz,
1 H), 5.05 (d, J = 11.9 Hz, 1 H), 5.55 (d, J = 5.0 Hz, 1 H, H-1), 7.25–
7.46 (m, 15 H).
13C NMR (100 MHz, CDCl3): δ = 24.5, 25.1, 26.1, 50.1, 64.2, 67.5,
69.8, 70.6, 70.9, 71.5, 71.8, 72.6, 73.7, 74.2, 74.9, 79.0, 81.9, 96.5,
104.7, 108.7, 109.4, 127.5, 127.6, 127.7, 127.9, 128.2, 128.4, 128.5,
128.7, 128.9, 138.2, 138.5, 139.0, 167.9.
2,3,4-Tri-O-benzyl-6-O-{4-[(5-dimethylaminonaphthalene-1-
sulfonylamino)methyl]-1H-1,2,3-triazol-1-yl}acetyl-β-D-gluco-
pyranosyl-(1→6)-1,2:3,4-di-O-isopropylidene-α-D-galactopyra-
nose (18)
From dansyl alkyne 8 (19 mg, 0.066 mmol) and azidoacetyl disac-
charide 14 (52 mg, 0.067 mmol), click reaction according to GP 3
and purification by column chromatography (EtOAc–PE, 1:4 to
5:5) led to 18 as a beige powder (69 mg, 98%); mp 94 °C (dec.);
[α]D20 –3.97 (c 0.5, CHCl3).
1H NMR (400 MHz, CDCl3): δ = 1.30, 1.31, 1.44, 1.48 (4 s, 12 H,
4 × CH3), 2.87 (s, 6 H, 2 × NCH3), 3.37–3.48 (m, 3 H), 3.63–3.73
(m, 2 H), 4.06–4.19 (m, 5 H), 4.24 (dd, J = 1.4, 7.8 Hz, 1 H), 4.31
(dd, J = 2.3, 5.0 Hz, 1 H), 4.40–4.45 (m, 2 H), 4.46 (d, J = 7.8 Hz,
1 H, H-1′), 4.59 (dd, J = 2.3, 7.8 Hz, 1 H), 4.72 (d, J = 11.0 Hz, 1
H), 4.76 (d, J = 11.0 Hz, 1 H), 4.79 (d, J = 10.5 Hz, 1 H), 4.88 (d,
J = 17.4 Hz, 1 H), 4.97–5.07 (m, 3 H), 5.52 (t, J = 6.0 Hz, 1 H, NH),
5.56 (d, J = 5.0 Hz, 1 H, H-1), 7.12–7.55 (m, 19 H), 8.21 (dd, J =
1.4, 7.4 Hz, 1 H), 8.25 (d, J = 8.7 Hz, 1 H), 8.52 (d, J = 8.7 Hz, 1 H).
13C NMR (100 MHz, CDCl3): δ = 24.5, 25.1, 26.1, 38.9, 45.5, 50.6,
64.7, 67.3, 69.9, 70.5, 70.8, 71.4, 72.4, 74.4, 74.8, 75.8, 76.8, 81.5,
84.5, 96.4, 104.4, 108.7, 109.5, 115.4, 118.8, 123.3, 123.8, 127.7,
127.8, 127.9, 128.0, 128.2, 128.3, 128.5, 128.6, 128.7, 129.7, 130.0,
130.7, 134.6, 137.7, 138.4, 138.6, 144.6, 152.1, 165.8.
HRMS (ESI): m/z calcd for C41H49N3O12 + Na [M + Na+]: 798.3214;
found: 798.3200.
2,3,4-Tri-O-benzyl-6-O-{4-[(7-nitrobenzo[1,2,5]oxadiazol-4-yl-
amino)methyl]-1H-1,2,3-triazol-1-yl}acetyl-β-D-glucopyrano-
syl-(1→6)-1,2:3,4-di-O-isopropylidene-α-D-galactopyranose (16)
From NBD alkyne 5 (29 mg, 0.132 mmol) and azidoacetyl disac-
charide 14 (107 mg, 0.138 mmol), click reaction according to GP 3
and purification by column chromatography (EtOAc–PE, 1:4 to
20
3:7) led to 16 as an orange powder (80 mg, 61%); mp 98 °C; [α]D
–39.1 (c 0.5, CHCl3).
1H NMR (400 MHz, CDCl3): δ = 1.30 (s, 6 H, CH3), 1.43 (s, 3 H,
CH3), 1.48 (s, 3 H, CH3), 3.36–3.42 (m, 2 H), 3.46–3.52 (m, 1 H),
© Georg Thieme Verlag Stuttgart · New York
Synthesis 2012, 44, 1647–1656

1654
H. Yi et al.
PAPER
HRMS (ESI): m/z calcd for C56H65N5O14S + Na [M + Na+]:
1086.4146; found: 1086.4157.
GP 3 and purification by column chromatography (EtOAc–PE, 1:4
to 5:5) led to 21 as a pink solid (60 mg, 89%); mp 113 °C (dec.);
[α]D20 –5.68 (c 0.62, CHCl3).
2,3,4-Tri-O-benzyl-6-O-{4-[(5-dimethylaminonaphthalene-1-
sulfonylamino)methyl]-1H-1,2,3-triazol-1-yl}acetyl-β-D-galac-
topyranosyl-(1→6)-1,2:3,4-di-O-isopropylidene-α-D-galactopy-
ranose (19)
From dansyl alkyne 8 (26 mg, 0.090 mmol) and azidoacetyl disac-
charide 15 (62 mg, 0.080 mmol), click reaction according to GP 3
and purification by column chromatography (EtOAc–PE, 1:4 to
5:5) led to 19 (65 mg, 76%) as a yellow/brown solid; mp 90 °C;
[α]D20 –98.2 (c 0.5, CHCl3).
1H NMR (400 MHz, CDCl3): δ = 1.30, 1.43, 1.48 (3 s, 12 H, 4 ×
CH3), 2.87 (s, 6 H, 2 × NCH3), 3.48–3.52 (m, 2 H, H-3,3′), 3.67 (dd,
J = 6.9, 9.6 Hz, 1 H), 3.70 (d, J = 2.3 Hz, 1 H), 3.83 (dd, J = 7.4, 9.6
Hz, 1 H, H-2′), 4.05–4.13 (m, 3 H), 4.18 (d, J = 6.4 Hz, 2 H), 4.21
(dd, J = 1.4, 7.8 Hz, 1 H), 4.27 (dd, J = 6.9, 11.0 Hz, 1 H), 4.30 (dd,
J = 2.3, 4.6 Hz, 1 H), 4.40 (d, J = 7.8 Hz, 1 H, H-1′), 4.57–4.58 (m,
1 H), 4.60 (d, J = 11.9 Hz, 1 H), 4.72 (d, J = 11.0 Hz, 1 H), 4.73 (d,
J = 11.9 Hz, 1 H), 4.83 (d, J = 11.9 Hz, 1 H), 4.87 (d, J = 6.9 Hz, 2
H, CH2), 4.93 (d, J = 11.9 Hz, 1 H), 5.05 (d, J = 11.0 Hz, 1 H), 5.55
(d, J = 4.6 Hz, 1 H, H-1), 5.56–5.63 (m, 1 H, NH), 7.16 (d, J = 7.3
Hz, 1 H), 7.25–7.53 (m, 18 H), 8.23–8.26 (m, 2 H), 8.52 (d, J = 8.2
Hz, 1 H).
1H NMR (400 MHz, CDCl3): δ = 1.13 (t, J = 6.9 Hz, 12 H, 4 × CH3),
1.30, 1.41, 1.43, 1.48 (4 s, 12 H, 4 × CH3), 3.29 (q, J = 6.9 Hz, 8 H,
4 × CH2), 3.37 (t, J = 6.4 Hz, 1 H), 3.50 (dd, J = 2.8, 9.6 Hz, 1 H,
H-3′), 3.64–3.68 (m, 2 H), 3.83 (dd, J = 7.8, 9.6 Hz, 1 H, H-2′),
4.02–4.13 (m, 3 H), 4.20–4.24 (m, 2 H), 4.30 (dd, J = 2.3, 4.6 Hz, 1
H, H-2), 4.40 (d, J = 7.8 Hz, 1 H, H-1′), 4.47 (s, 2 H, OCH2), 4.58
(dd, J = 2.3, 5.5 Hz, 1 H), 4.59 (d, J = 11.9 Hz, 1 H, CHPh), 4.71–
4.76 (m, 4 H), 4.85 (d, J = 11.9 Hz, 1 H, CHPh), 4.92 (d, J = 11.9
Hz, 1 H, CHPh), 5.06 (d, J = 11.0 Hz, 1 H, CHPh), 5.55 (d, J = 5.0
Hz, 1 H, H-1), 6.13–6.15 (m, 2 H), 6.29 (dd, J = 3.2, 8.7 Hz, 2 H,
ArH), 6.33–6.35 (m, 2 H), 7.07–7.09 (m, 1 H, ArH), 7.25–7.44 (m,
18 H, ArH), 7.90–7.93 (m, 1 H, CH=).
13C NMR (100 MHz, CDCl3): δ = 12.7, 24.5, 25.1, 26.1, 27.0, 35.4,
44.4, 50.2, 64.5, 65.1, 67.5, 70.9, 71.5, 73.7, 74.2, 74.9, 78.9, 81.8,
87.5, 96.5, 97.9, 104.7, 105.1, 108.0, 108.7, 109.4, 123.0, 123.9,
124.0, 128.1, 128.2, 128.5, 128.7, 128.8, 130.9, 132.7, 138.0, 138.6,
139.0, 145.0, 148.8, 153.4, 153.8, 165.6.
HRMS (ESI): m/z calcd for C72H82N6O14S + Na [M + Na+]:
1277.5787; found: 1277.5680.
13C NMR (100 MHz, CDCl3): δ = 24.5, 25.1, 26.1, 38.8, 45.5, 50.5,
64.8, 67.4, 69.7, 70.6, 70.8, 71.5, 71.7, 72.7, 73.7, 74.2, 74.9, 79.0,
81.8, 96.5, 104.6, 108.7, 109.5, 115.4, 118.8, 123.3, 123.9, 127.5,
127.6, 127.7, 128.0, 128.2, 128.4, 128.5, 128.6, 128.7, 128.8, 129.6,
129.7, 130.0, 130.7, 134.6, 138.2, 138.5, 139.0, 144.6, 152.1, 165.8.
O-[2,3,4-Tri-O-benzyl-6-O-(2-azido)acetyl-D-glucopyranosyl]-
N-benzyloxycarbonyl-L-serine Benzyl Ester (22)
Compound 3 (314 mg, 0.502 mmol) was treated with N-Z-L-Ser-
OBn (176 mg, 0.534 mmol) according to GP 2. Purification by
chromatography (EtOAc–PE, 1:9 to 3:7) gave 330 mg (78%) of 22
as a mixture of β/α (3:1) anomers; white solid.
HRMS (ESI): m/z calcd for C56H65N5O14S + Na [M + Na+]:
1086.4146; found: 1086.4135.
β-Anomer
1H NMR (400 MHz, CDCl3): δ = 3.30–3.46 (m, 4 H), 3.60–3.88 (m,
4 H), 4.18–4.40 (m, 3 H), 4.34 (d, J = 7.8 Hz, 1 H, H-1), 4.53–4.97
(m, 6 H), 5.08–5.20 (m, 4 H), 5.66–5.68 (d, J = 8.2 Hz, 1 H, NH),
7.20–7.33 (m, 25 H).
2,3,4-Tri-O-benzyl-6-O-[2-(4-{[3′,6′-bis(diethylamino)-3-oxo-
spiro(isoindoline-1,9′-xanthene)-2-yl]methyl}-1H-1,2,3-triazol-
1-yl)]acetyl-β-D-glucopyranosyl-(1→6)-1,2:3,4-di-O-isopropyli-
dene-α-D-galactopyranose (20)
From rhodamine alkyne 11 (53 mg, 0.110 mmol) and azidoacetyl
disaccharide 14 (85 mg, 0.110 mmol), click reaction according to
GP 3 and purification by column chromatography (EtOAc–PE, 1:4
to 5:5) led to 20 (137 mg, 99%) as a red solid; mp 88 °C; [α]D20 –7.5
(c 0.5, CHCl3).
13C NMR (100 MHz, CDCl3): δ = 50.0, 54.5, 64.1, 67.1, 67.4, 67.7,
72.6, 74.9, 75.0, 75.7, 76.7, 76.8, 81.8, 84.5, 103.7, 127.8, 127.9,
128.0, 128.1, 128.2, 128.3, 128.5, 128.6, 135.3, 136.2, 137.6, 138.0,
138.3, 156.0, 168.1, 169.8.
HRMS (ESI): m/z calcd for C47H48N4O11 + Na [M + Na+]: 867.3217;
1H NMR (400 MHz, CDCl3): δ = 1.23 (t, J = 6.9 Hz, 12 H, 4 × CH3),
1.30, 1.31, 1.43, 1.48 (4 s, 12 H, 4 × CH3), 3.28 (q, J = 6.9 Hz, 8 H,
4 × CH2), 3.38–3.42 (m, 3 H), 3.64 (t, J = 9.2 Hz, 1 H, H-3′), 3.68–
3.73 (m, 1 H), 4.05–4.13 (m, 2 H), 4.16 (dd, J = 5.0, 11.9 Hz, 1 H),
4.23 (dd, J = 1.4, 7.8 Hz, 1 H), 4.31 (dd, J = 2.8, 5.0 Hz, 1 H, H-2),
4.37 (dd, J = 1.8, 11.4 Hz, 1 H), 4.44 (d, J = 7.8 Hz, 1 H, H-1′), 4.47–
4.49 (m, 3 H), 4.59 (dd, J = 2.3, 7.8 Hz, 1 H), 4.70 (d, J = 11.0 Hz,
1 H), 4.75 (d, J = 11.0 Hz, 1 H), 4.80–4.87 (m, 3 H), 4.97 (d, J =
10.6 Hz, 1 H), 5.04 (d, J = 11.0 Hz, 1 H), 5.55 (d, J = 4.4 Hz, 1 H,
H-1), 6.14–6.15 (m, 2 H), 6.27–6.33 (m, 4 H), 7.08–7.43 (m, 19 H),
7.90–7.91 (m, 1 H).
13C NMR (100 MHz, CDCl3): δ = 12.7, 24.5, 25.1, 26.1, 31.0, 35.3,
44.4, 50.1, 64.6, 65.1, 67.4, 70.5, 70.8, 71.5, 72.5, 74.4, 74.9, 75.7,
81.4, 84.5, 96.4, 97.9, 104.4, 105.3, 108.0, 108.6, 109.5, 122.9,
123.9, 124.1, 127.6, 127.7, 128.0, 128.2, 128.3, 128.5, 128.6, 128.7,
128.8, 132.6, 137.7, 138.5, 138.6, 144.9, 148.8, 153.4, 166.0, 168.0.
found: 867.3206.
O-[2,3,4-Tri-O-benzyl-6-O-(2-azido)acetyl-D-galactopyrano-
syl]-N-benzyloxycarbonyl-L-serine Benzyl Ester (23)
Compound 4 (202 mg, 0.323 mmol) was treated with N-Z-L-Ser-
OBn (112 mg, 0.340 mmol) according to GP 2. Purification by
chromatography (EtOAc–PE, 1:9 to 1:3) gave 204 mg (75%) of 23
(β/α = 9:1) as a beige solid.
1H NMR (400 MHz, CDCl3): δ = 3.48–3.53 (m, 2 H), 3.70 (s, 2 H,
CH2), 3.77–3.82 (m, 2 H), 3.83 (dd, J = 2.3, 10.6 Hz, 1 H), 4.13 (dd,
J = 6.0, 11.0 Hz, 1 H), 4.24–4.29 (m, 1 H), 4.30 (d, J = 7.8 Hz, 1 H),
4.38 (dd, J = 2.3, 10.1 Hz, 1 H), 4.57–4.59 (m, 1 H), 4.65 (d, J =
11.5 Hz, 1 H), 4.70 (d, J = 11.0 Hz, 1 H), 4.76 (d, J = 11.4 Hz, 2 H),
4.82 (d, J = 11.9 Hz, 1 H), 4.99 (d, J = 11.5 Hz, 1 H), 5.06 (d, J =
12.3 Hz, 1 H), 5.10 (d, J = 12.3 Hz, 1 H), 5.19 (d, J = 12.4 Hz, 1 H),
5.23 (d, J = 12.4 Hz, 1 H), 5.77 (d, J = 7.8 Hz, 1 H, NH), 7.18–7.40
(m, 25 H).
13C NMR (100 MHz, CDCl3): δ = 50.1, 54.6, 64.3, 67.1, 67.5, 70.0,
72.1, 72.9, 73.6, 74.5, 75.5, 75.9, 79.0, 82.0, 104.3, 127.7, 127.9,
128.1, 128.2, 128.4, 128.5, 128.6, 128.7, 135.4, 136.4, 138.1, 138.2,
138.3, 156.1, 168.0, 169.8.
HRMS (ESI): m/z calcd for C47H48N4O11 + Na [M + Na+]: 867.3217;
found: 867.3211.
HRMS (ESI): m/z calcd for C72H82N6O14S + Na [M + Na+]:
1277.5787; found: 1277.5562.
2,3,4-Tri-O-benzyl-6-O-[2-(4-{[(3′,6′-bis(diethylamino)-3-oxo-
spiro(isoindoline-1,9′-xanthene)-2-yl)methyl]-1H-1,2,3-triazol-
1-yl)]acetyl-β-D-galactopyranosyl-(1→6)-1,2:3,4-di-O-isopro-
pylidene-α-D-galactopyranose (21)
From rhodamine alkyne 11 (28 mg, 0.058 mmol) and azidoacetyl
disaccharide 15 (42 mg, 0.054 mmol), click reaction according to
Synthesis 2012, 44, 1647–1656
© Georg Thieme Verlag Stuttgart · New York

PAPER
Synthesis of Triazole-Linked Fluorescent Saccharides
1655
O-{2,3,4-Tri-O-benzyl-6-O-{4-[(5-dimethylaminonaphthalene-
1-sulfonylamino)methyl]-1H-1,2,3-triazol-1-yl}acetyl-D-gluco-
pyranosyl}-N-benzyloxycarbonyl-L-serine Benzyl Ester (24)
From dansyl alkyne 8 (15 mg, 0.052 mmol) and azidoacetyl gluco-
syl amino ester 22 (44 mg, 0.052 mmol), click reaction according to
GP 3 and purification by column chromatography (EtOAc–PE, 1:4
to 5:5) led to 24 (β/α = 3:1) as a beige solid (55 mg, 93%).
HRMS (ESI): m/z calcd for C78H81N7O13 + Na [M + Na+]:
1346.5790; found: 1346.5746.
O-{2,3,4-Tri-O-benzyl-6-O-[2-(4-{[(3′,6′-bis(diethylamino)-3-
oxospiro(isoindoline-1,9′-xanthene)-2-yl]methyl}-1H-1,2,3-tri-
azol-1-yl)]acetyl-D-galactopyranosyl}-N-benzyloxycarbonyl-L-
serine Benzyl Ester (27)
From rhodamine alkyne 11 (21 mg, 0.044 mmol) and azidoacetyl
glucosyl amino ester 23 (37 mg, 0.044 mmol), click reaction ac-
cording to GP 3 and purification by column chromatography
(EtOAc–PE, 5:5) led to 27 as a pink solid (53 mg, 91%) (β/α = 9:1).
1H NMR (400 MHz, CDCl3): δ = 1.13 (t, J = 6.9 Hz, 12 H, 4 × CH3),
3.29 (q, J = 6.9 Hz, 8 H, 4 × CH2), 3.41 (t, J = 6.0 Hz, 1 H, H-5),
3.46 (dd, J = 2.3, 9.6 Hz, 1 H, H-3), 3.66 (m, 1 H, H-4), 3.74 (dd,
J = 7.8, 9.6 Hz, 1 H, H-2), 3.82 (dd, J = 3.2, 10.6 Hz, 1 H, OCHSer),
4.02 (dd, J = 5.5, 11.0 Hz, 1 H, H-6), 4.20 (dd, J = 6.4, 11.0 Hz, 1
H, H-6′), 4.27 (d, J = 7.8 Hz, 1 H, H-1), 4.35 (dd, J = 2.8, 10.1 Hz,
1 H, OCHSer), 4.45 (s, 2 H, OCH2), 4.55 (d, J = 11.9 Hz, 1 H, CHPh),
4.56–4.57 (m, 1 H, NCH), 4.67 (d, J = 11.0 Hz, 1 H, CHPh), 4.72–
4.79 (m, 4 H), 4.94 (d, J = 11.9 Hz, 1 H, CHPh), 4.92 (d, J = 11.9
Hz, 1 H, CHPh), 5.02–5.10 (m, 2 H), 5.14–5.22 (m, 2 H), 5.80 (d,
J = 8.2 Hz, 1 H, NH), 6.12–6.14 (m, 2 H, ArH), 6.28–6.34 (m, 4 H,
ArH), 7.07–7.09 (m, 1 H, ArH), 7.25–7.31 (m, 28 H, ArH), 7.89–
7.91 (m, 1 H, CH=).
β-Anomer
1H NMR (400 MHz, CDCl3): δ = 2.87 (s, 6 H, 2 × CH3), 3.30–3.36
(m, 3 H), 3.59 (t, J = 9.2 Hz, 1 H), 3.88 (dd, J = 3.6, 10.5 Hz, 1 H),
4.08–4.18 (m, 4 H), 4.31 (d, J = 7.8 Hz, 1 H, H-1), 4.35–4.38 (m, 1
H), 4.45–4.48 (m, 1 H), 4.56–4.67 (m, 2 H), 4.75–4.94 (m, 6 H),
5.07–5.18 (m, 4 H), 5.67–5.69 (m, 1 H, NH), 5.79 (d, J = 7.8 Hz, 1
H, NH), 7.15–7.55 (m, 29 H), 8.21 (dd, J = 0.9, 7.3 Hz, 1 H), 8.27
(d, J = 8.3 Hz, 1 H), 8.52 (d, J = 8.3 Hz, 1 H).
13C NMR (100 MHz, CDCl3): δ = 38.9, 45.5, 50.6, 54.7, 60.5, 64.6,
67.2, 67.6, 69.6, 72.5, 74.9, 75.0, 75.8, 77.3, 81.8, 84.4, 103.6,
115.4, 118.9, 123.3, 123.9, 127.8, 127.9, 128.0, 128.1, 128.2, 128.5,
128.6, 128.7, 129.7, 130.7, 134.6, 135.3, 136.2, 137.6, 138.1, 138.2,
144.7, 152.0, 156.1, 165.8, 170.0.
HRMS (ESI): m/z calcd for C62H64N6O13S + Na [M + Na+]:
1155.4150; found: 1155.4149.
13C NMR (100 MHz, CDCl3): δ = 12.7, 29.8, 35.4, 44.4, 50.2, 54.6,
64.5, 65.2, 67.1, 67.5, 70.0, 71.9, 72.8, 73.5, 74.5, 79.0, 82.0, 98.0,
104.2, 105.2, 108.0, 123.0, 123.9, 124.2, 128.6, 128.7, 132.7, 135.3,
138.1, 138.2, 145.0, 148.8, 153.5, 153.7, 156.1, 165.8, 168.0, 169.9.
HRMS (ESI): m/z calcd for C78H81N7O13 + Na [M + Na+]:
1346.5790; found: 1346.5567.
O-{2,3,4-Tri-O-benzyl-6-O-{4-[(5-dimethylaminonaphthalene-
1-sulfonylamino)methyl]-1H-1,2,3-triazol-1-yl}acetyl-D-galac-
topyranosyl}-N-benzyloxycarbonyl-L-serine Benzyl Ester (25)
From dansyl alkyne 8 (14 mg, 0.048 mmol) and azidoacetyl gluco-
syl amino ester 23 (43 mg, 0.051 mmol), click reaction according to
GP 3 and purification by column chromatography (EtOAc–PE, 1:4
to 5:5) led to 25 (44 mg, 76%) (β/α = 9:1) as a beige solid.
1H NMR (400 MHz, CDCl3): δ = 2.87 (s, 6 H, 2 × CH3), 3.42–3.50
(m, 2 H), 3.67 (s, 2 H, CH2), 3.72–3.78 (m, 2 H), 3.84 (dd, J = 3.7,
10.5 Hz, 1 H), 3.99 (dd, J = 4.6, 11.0 Hz, 1 H), 4.15–4.29 (m, 4 H),
4.57 (d, J = 11.9 Hz, 2 H), 4.65–5.20 (m, 10 H), 5.57 (t, J = 6.0 Hz,
1 H, NH), 5.82 (d, J = 7.8 Hz, 1 H, NH), 7.12–7.52 (m, 29 H), 8.21–
8.26 (m, 2 H), 8.52 (d, J = 8.7 Hz, 1 H).
13C NMR (100 MHz, CDCl3): δ = 38.9, 45.6, 50.7, 54.7, 65.1, 67.2,
67.6, 69.9, 72.0, 73.1, 73.6, 74.5, 75.4, 79.0, 81.9, 104.2, 115.5,
118.8, 123.3, 124.0, 127.8, 127.9, 128.2, 128.5, 128.6, 128.7, 129.8,
130.0, 130.7, 134.6, 135.3, 136.3, 138.1, 138.3, 138.4, 144.8, 152.1,
156.2, 165.8, 170.0.
3-[6-O-(2-Azido)acetyl-2,3,4-tri-O-benzyl-α-D-glucopyrano-
syl]prop-1-ene (29)
α-C-Allyl glucopyranoside 28 (259 mg, 0.546 mmol) was treated
successively with ClCH2COCl and NaN3 according to GP 1. Purifi-
cation by column chromatography (EtOAc–PE, 1:9) gave 213 mg
(70%) of compound 29 as a white solid; mp 72 °C; [α]D20 +54.9 (c,
0.5 CHCl3).
1H NMR (400 MHz, CDCl3): δ = 2.46 (t, J = 6.9 Hz, 2 H, CH2), 3.41
(dd, J = 8.7, 10.1 Hz, 1 H, H-4), 3.67–3.69 (m, 1 H, H-5), 3.71 (dd,
J = 5.5, 9.2 Hz, 1 H, H-2), 3.73 (d, J = 17.4 Hz, 1 H), 3.79 (d, J =
17.4 Hz, 1 H), 3.82 (t, J = 9.2 Hz, 1 H, H-3), 4.02–4.08 (m, 1 H, H-
1), 4.24 (dd, J = 5.0, 11.4 Hz, 1 H, H-6), 4.31 (dd, J = 2.3, 11.4 Hz,
1 H, H-6′), 4.56 (d, J = 11.0 Hz, 1 H), 4.61 (d, J = 11.4 Hz, 1 H),
4.68 (d, J = 11.9 Hz, 1 H), 4.79 (d, J = 11.0 Hz, 1 H), 4.86 (d, J =
11.0 Hz, 1 H), 4.94 (d, J = 10.5 Hz, 1 H), 5.05–5.11 (m, 2 H, CH2=),
5.69–5.75 (m, 1 H, CH=), 7.25–7.35 (m, 15 H).
13C NMR (100 MHz, CDCl3): δ = 30.0, 50.2, 64.8, 69.5, 73.3, 73.7,
75.1, 75.6, 77.4, 80.1, 82.3, 117.3, 128.0, 128.1, 128.2, 128.3,
128.6, 128.7, 134.5, 137.8, 138.1, 138.6, 168.2.
HRMS (ESI): m/z calcd for C32H35N3O6 + Na [M + Na+]: 580.2424;
found: 580.2435.
HRMS (ESI): m/z calcd for C62H64N6O13S + Na [M + Na+]:
1155.4150; found: 1155.4142.
O-{2,3,4-Tri-O-benzyl-6-O-[2-(4-{[3′,6′-bis(diethylamino)-3-
oxospiro(isoindoline-1,9′-xanthene)-2-yl]methyl}-1H-1,2,3-tri-
azol-1-yl)]acetyl-D-glucopyranosyl}-N-benzyloxycarbonyl-L-
serine Benzyl Ester (26)
From rhodamine alkyne 11 (33 mg, 0.069 mmol) and azidoacetyl
glucosyl amino ester 22 (58 mg, 0.069 mmol), click reaction ac-
cording to GP 3 and purification by column chromatography (EtO-
Ac–PE, 1:4 to 5:5) led to 26 (89 mg, 98%) (β/α = 3:1) as a red solid.
1H NMR (400 MHz, CDCl3): δ = 1.10–1.14 (m, 12 H, 4 × CH3),
3.24–3.34 (m, 11 H), 3.59 (t, J = 8.7 Hz, 0.75 H, H-3β), 3.70 (t, J =
6.9 Hz, 0.25 H, H-3α), 3.81–3.83 (m, 0.25 H), 3.88 (dd, J = 3.7, 10.6
Hz, 0.75 H), 3.88–4.20 (m, 1 H), 4.33 (d, J = 7.8 Hz, 0.75 H, H-1β),
4.29–4.37 (m, 1.25 H), 4.46–4.54 (m, 3 H), 4.57–4.62 (m, 3 H),
4.73–4.93 (m, 6 H), 5.02–5.19 (m, 4 H), 5.85 (d, J = 7.8 Hz, 1 H,
NH), 6.11–6.13 (m, 2 H, ArH), 6.27–6.34 (m, 4 H, ArH), 7.07 (d,
J = 6.4 Hz, 1 H, ArH), 7.19–7.52 (m, 28 H, ArH), 7.91 (d, J = 6.4
Hz, 1 H, CH=).
13C NMR (100 MHz, CDCl3): δ = 12.7, 35.4, 44.4, 50.2, 54.7, 60.5,
64.5, 65.1, 67.1, 67.4, 69.7, 72.6, 74.9, 75.0, 75.7, 77.3, 81.2, 84.5,
98.0, 103.6, 105.3, 108.0, 123.0, 123.9, 124.2, 127.9, 128.0, 128.2,
128.5, 128.6, 128.7, 128.8, 130.9, 132.6, 135.4, 136.3, 137.5, 138.1,
138.3, 144.9, 148.8, 153.5, 156.1, 166.0, 168.0, 169.9.
3-{2,3,4-Tri-O-benzyl-6-O-{4-[(5-dimethylaminonaphthalene-
1-sulfonylamino)methyl]-1H-1,2,3-triazol-1-yl}acetyl-α-D-glu-
copyranosyl}prop-1-ene (31)
From dansyl alkyne 8 (15 mg, 0.052 mmol) and azidoacetyl C-glu-
coside 29 (29 mg, 0.052 mmol), click reaction according to GP 3
and purification by column chromatography (EtOAc–PE, 1:9 to
1:4) led to 31 (41 mg, 93%).
1H NMR (400 MHz, CDCl3): δ = 2.43–2.47 (m, 2 H, CH2), 2.87 (s,
6 H, 2 × CH3), 3.30 (dd, J = 8.2, 9.6 Hz, 1 H, H-4), 3.65 (ddd, J =
2.2, 4.6, 10.1 Hz, 1 H, H-5), 3.72 (dd, J = 5.5, 9.2 Hz, 1 H, H-2),
3.81 (t, J = 9.2 Hz, 1 H, H-3), 4.04–4.09 (m, 1 H, H-1), 4.15–4.19
(m, 3 H, H-6, NCH2), 4.33 (dd, J = 2.3, 11.9 Hz, 1 H, H-6′), 4.45 (d,
J = 11.4 Hz, 1 H), 4.62 (d, J = 11.4 Hz, 1 H), 4.71 (d, J = 10.2 Hz,
1 H), 4.78 (d, J = 11.0 Hz, 1 H), 4.80 (d, J = 11.0 Hz, 1 H), 4.87–
4.99 (m, 5 H), 5.06–5.10 (m, 2 H, CH2=), 5.43 (t, J = 6.4 Hz, 1 H,
© Georg Thieme Verlag Stuttgart · New York
Synthesis 2012, 44, 1647–1656

1656
H. Yi et al.
PAPER
C. Bioorg. Med. Chem. 2010, 18, 7439. (c) Oka, H.;
Koyama, T.; Hatano, K.; Terunuma, D.; Matsuoka, K.
Bioorg. Med. Chem. Lett. 2010, 20, 1969.
NH), 5.70–5.78 (m, 1 H, CH=), 7.23–7.54 (m, 18 H), 8.20 (dd, J =
1.4, 7.6 Hz, 2 H), 8.23 (d, J = 8.3 Hz, 1 H), 8.52 (d, J = 8.7 Hz, 1 H).
13C NMR (100 MHz, CDCl3): δ = 30.0, 39.0, 50.7, 65.2, 69.4, 73.2,
73.6, 75.0, 75.6, 77.4, 79.9, 82.2, 115.5, 117.3, 118.8, 123.3, 123.9,
127.9, 128.0, 128.1, 128.2, 128.3, 128.6, 128.7, 128.8, 129.7, 130.1,
130.8, 134.5, 134.6, 137.8, 138.1, 138.4, 144.6, 152.1, 167.8.
(6) Gege, C.; Oscarson, S.; Schmidt, R. R. Tetrahedron Lett.
2001, 42, 377.
(7) Tian, Y. S.; Lee, H. Y.; Lim, C. S.; Park, J.; Kim, H. M.;
Shin, Y. N.; Kim, E. S.; Jeon, H. J.; Park, S. B.; Cho, B. R.
Angew. Chem. Int. Ed. 2009, 48, 8027.
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S. B.; Peters, E. C.; Agnew, B. J.; Hsieh-Wilson, L. C. J. Am.
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Samanta, A.; Yun, S.-W.; Chang, Y.-T. Org. Biomol. Chem.
2011, 9, 4760.
HRMS (ESI): m/z calcd for C47H51N5O8S + Na [M + Na+]:
868.3356; found: 868.3390.
3-{2,3,4-Tri-O-benzyl-6-O-[2-(4-{[(3′,6′-bis(diethylamino)-3-
oxospiro(isoindoline-1,9′-xanthene)-2-yl]methyl}-1H-1,2,3-tri-
azol-1-yl)]acetyl-α-D-glucopyranosyl}prop-1-ene (32)
From rhodamine alkyne 11 (19 mg, 0.040 mmol) and azidoacetyl C-
glucoside 29 (30 mg, 0.054 mmol), click reaction according to GP
3 and purification by column chromatography (EtOAc–PE, 2:8 to
5:5) led to 32 as a red solid (55 mg, 99%); mp 81 °C; [α]D20 +57.8
(c, 0.5 CHCl3).
(10) Katritzky, A. R.; Cusido, J.; Narindoshvili, T. Bioconjugate
Chem. 2008, 19, 1471.
1H NMR (400 MHz, CDCl3): δ = 1.12 (m, 12 H, 4 × CH3), 2.43–
2.46 (m, 2 H, CH2), 3.25–3.35 (m, 9 H, 4 × CH2, H-4), 3.65 (ddd,
J = 2.2, 4.6, 10.1 Hz, 1 H, H-5), 3.69 (dd, J = 5.5, 9.6 Hz, 1 H, H-
2), 3.79 (t, J = 8.7 Hz, 1 H, H-3), 4.01–4.08 (m, 1 H, H-1), 4.14 (dd,
J = 5.5, 11.4 Hz, 1 H, H-6), 4.29 (dd, J = 2.3, 11.9 Hz, 1 H, H-6′),
4.48–4.51 (m, 3 H), 4.62 (d, J = 11.4 Hz, 1 H), 4.68 (d, J = 11.4 Hz,
1 H), 4.76–4.84 (m, 4 H), 4.93 (d, J = 11.0 Hz, 1 H), 5.05–5.10 (m,
2 H, CH2=), 5.65–5.73 (m, 1 H, CH=), 6.12–6.15 (m, 2 H), 6.27–
6.33 (m, 4 H), 7.06–7.45 (m, 19 H), 7.90–7.93 (m, 1 H).
13C NMR (100 MHz, CDCl3): δ = 12.7, 30.0, 35.3, 44.4, 50.2, 65.0,
69.4, 73.2, 73.5, 74.9, 75.4, 77.4, 79.9, 82.1, 97.9, 105.3, 108.0,
117.2, 123.0, 123.9, 124.1, 127.8, 127.9, 128.0, 128.2, 128.3, 128.6,
128.7, 128.9, 130.9, 132.6, 134.4, 137.7, 138.1, 138.5, 144.8, 148.8,
153.4, 153.7, 166.0, 168.0.
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Aust. J. Chem. 2004, 57, 885.
(13) Nagarajan, S.; Das, T. M. New J. Chem. 2009, 33, 2391.
(14) Renaudet, O.; Dumy, P. Eur. J. Org. Chem. 2008, 5383.
(15) (a) Rostovtsev, V. V.; Green, L. G.; Fokin, V. V.; Sharpless,
K. B. Angew. Chem. Int. Ed. 2002, 41, 2596. (b) Tornoe, C.
W.; Christensen, C.; Meldal, M. J. Org. Chem. 2002, 67,
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A.; Shavyrin, A. S.; Lopatin, M. A.; Balalaeva, I. V.;
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2007, 48, 6527. (b) Maisonneuve, S.; Fang, J.; Xie, J.
Tetrahedron 2008, 64, 8716. (c) Lin, L.; Shen, Q.; Chen, Q.-
R.; Xie, J. Bioorg. Med. Chem. 2008, 16, 9757. (d) Cheng,
K. G.; Liu, J.; Liu, X. F.; Li, H. L.; Sun, H. B.; Xie, J.
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Synlett 2009, 2977. (g) Cheng, K.; Liu, J.; Sun, H.; Xie, J.
Synthesis 2010, 1046. (h) Li, C.; Henry, E.; Mani, N. K.;
Tang, J.; Brochon, J.-C.; Deprez, E.; Xie, J. Eur. J. Org.
Chem. 2010, 2395. (i) Garcia, L.; Maisonneuve, S.; Xie, J.;
Guillot, R.; Dorlet, P.; Rivière, E.; Desmadril, M.; Lambert,
F.; Policar, C. Inorg. Chem. 2010, 49, 7282. (j) Garcia, L.;
Maisonneuve, S.; Oudinet-Sin Marcu, J.; Guillot, R.;
Lambert, F.; Xie, J.; Policar, C. Inorg. Chem. 2011, 50,
11353.
HRMS (ESI): m/z calcd for C63H68N6O8S + Na [M + Na+]:
1059.4996; found: 1127.4833.
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Synthesis 2012, 44, 1647–1656
© Georg Thieme Verlag Stuttgart · New York