Glyco-SAMs by 'dual click': Thiourea-bridged glyco-OEG azides for cycloaddition on surfaces

Article abstract of DOI:10.1055/S-0029-1218611

A series of NCS-functionalized sugars were synthesized and used in a thiourea-bridging reaction with aminohexa(ethylene glycol) azide [H ₂N(EG)₆N₃], a bifunctional oligo(ethylene glycol) derivative, which can be used as key intermediate for the fabrication of biorepulsive glyco-SAMs by a 'dual click' approach. Glyco-SAMs can serve as defined glycocalyx models for the study of carbohydrate-protein interactions. The copper(I)-catalyzed 1,3-dipolar cycloaddition reaction of the obtained glyco-OEG azides was exemplified, which can be used to modify preformed monolayers 'on SAM'. Georg Thieme Verlag Stuttgart.

Full text of DOI:10.1055/S-0029-1218611

828
PAPER
Glyco-SAMs by ‘Dual Click’: Thiourea-Bridged Glyco-OEG Azides for
Cycloaddition on Surfaces
G
lyco-SAMs by ‘
a
D
ual Click
r
’
sten Grabosch,a Mike Kleinert,b Thisbe K. Lindhorst*a
a
Otto Diels Institute of Organic Chemistry, Christiana Albertina University of Kiel, Otto-Hahn Platz 3, 24098 Kiel, Germany
Fax +49(431)8807410; E-mail: tklind@oc.uni-kiel.de
b
Department of Chemistry, University of Bergen, Allégaten 41, 5007 Bergen, Norway
Received 5 October 2009; revised 12 November 2009
models are needed, which allow defined variation of the
structural set-up of a carbohydrate-coated surface.2 An
ideal system, which offers a reliable as well as flexible
construction of ordered glycoarrays are self-assembled
Abstract: A series of NCS-functionalized sugars were synthesized
and used in a thiourea-bridging reaction with aminohexa(ethylene
glycol) azide [H2N(EG)6N3], a bifunctional oligo(ethylene glycol)
derivative, which can be used as key intermediate for the fabrication
of biorepulsive glyco-SAMs by a ‘dual click’ approach. Glyco- monolayers, in short SAMs.3 SAMs can be functionalized
with carbohydrate head groups to obtain ‘glyco-SAMs’,4
SAMs can serve as defined glycocalyx models for the study of car-
bohydrate-protein interactions. The copper(I)-catalyzed 1,3-dipolar
which allow the study of carbohydrate-protein interac-
cycloaddition reaction of the obtained glyco-OEG azides was ex-
tions using different biophysical methods.5
emplified, which can be used to modify preformed monolayers ‘on
Self-assembled monolayers are typically formed with
OEG[oligo(ethylene glycol)]-substituted alkane thiols,
which chemisorb onto a gold surface (Scheme 1). The al-
kane thiols that form the monolayer have to be attached to
an OEG chain, typically with EG3 to EG7, to resist unspe-
cific adsorption of proteins.6 Protein-repelling properties
of the unfunctionalized SAM are a prerequisite for em-
ployment of glyco-SAMs in carbohydrate-protein binding
studies. Finally, for glyco-SAM construction, molecules
of the type HO(CH2CH2O)n(CH2)11SH have to be func-
tionalized with a carbohydrate head group for carbohy-
drate-recognition studies.
SAM’.
Key words: carbohydrates, self-assembled monolayers, ‘click’
chemistry, thiourea-bridging
Every eukaryotic cell is covered by a highly complex sug-
ar coat, termed ‘glycocalyx’, comprised of a variety of
complex glycoconjugates. These carbohydrates are of
fundamental importance for cell-cell recognition and cell
adhesion.1 To study the details of the molecular recogni-
tion processes, that occur on the cell surface, synthetic
HN
SCN
thiourea
linkage
S
'click'
NH2
HN
carbohydrate
head group
O
employed
carbohydrate isothiocyanates
O
O
O
O
O
(OAc)n
O
O
O
OEG-chain
O
O
O
O
NCS
N
(OAc)n
O
'click'
N
N3
N
triazole
linkage
O
O
O
O
alkane thiol
O
(OAc)n
O
NCS
O
S
S
S
S
S
S
NCS
n ≥ 1
Au
Au
Au
Scheme 1 Construction of glyco-SAMs has to combine the required alkane thiols with protein-repulsive OEG chains and biorelevant carbo-
hydrate head groups. This is achieved in a chemoselective ‘dual click’ approach employing an a-amino-w-azido-difunctionalized OEG
[H2N(EG)6N3] in 1,3-dipolar addition with alkynes at one end and thiourea-bridging with isothiocyanato-functionalized sugars at the other.
Three different types of NCS-functionalized O-acetylated sugars were used in this study.
SYNTHESIS 2010, No. 5, pp 0828–0836
x
x
.
x
x
.2
0
1
0
Advanced online publication: 22.12.2009
DOI: 10.1055/s-0029-1218611; Art ID: T19409SS
© Georg Thieme Verlag Stuttgart · New York

PAPER
Glyco-SAMs by ‘Dual Click’
829
OAc
OAc
Glyco-SAMs can be constructed in two different ways: (i)
either the completely functionalized thiol is synthesized
first and subsequently assembled on the gold substrate; or
(ii) a basic monolayer is formed first and this blank is then
further refined ‘on SAM’. The latter concept enables
greater flexibility for biological testing, because a partic-
ular SAM can be investigated before and after further
modification. Thus, ‘switching’ the condition of a mono-
layer may allow to draw conclusions about the biological
consequences of a specific change, which is not possible
otherwise.
OAc
OAc
O
AcO
AcO
a
OAc
O
AcO
AcO
1
HO
Br
60%
O
R
3: R = Br
b, 94%
c, 80%
4: R = N3
5: R = NCS
AcO
AcO
AcO
OAc
O
We have recently reported on a ‘click on SAM’ ap-
proach,7 which relies on copper(I)-catalyzed 1,3-dipolar
cycloaddition of azides to alkyne-terminated SAMs.8 This
approach can be extended to a ‘dual click’ concept, in
which two essential steps for SAM fabrication are simpli-
fied, both OEG functionalization of the alkane thiol as
well as the sugar decoration (Scheme 1). The difunction-
alized OEG, aminohexa(ethylene glycol) azide
[H2N(EG)6N3], is the key molecule in this approach which
allows 1,3-dipolar addition to alkynes on one hand and the
well established thiourea-bridging reaction to
isothiocyanates9 on the other. The sequence of these two
operations can be altered according to requirements. In
this paper, the facile synthesis of NCS-functionalized gly-
cosides and thiourea-bridging to H2N(EG)6N3 is shown
and copper(I)-catalyzed cycloaddition to a long-chain
alkyne is exemplified.
2
O
CCl3
OAc
NH
HO
Br
d
n
n = 5 77%
n = 9 76%
OAc
O
AcO
AcO
O
R
n
6: n = 5, R = Br
7: n = 9, R = Br
e
e
n = 5 73%
n = 9 85%
8: n = 5, R = N3
9: n = 9, R = N3
f
f
n = 5 58%
n = 9 56%
10: n = 5, R = NCS
11: n = 9, R = NCS
Three types of NCS-functionalized carbohydrate deriva-
Scheme 2 Reagents and conditions: (a) BF3·OEt2, CH2Cl2, r.t., 12
tives were used. Glycosyl isothiocyanates carrying the h; (b) NaN3, Bu4NBr, DMF, 60 °C, 5 h, r.t. 12 h; (c) CS2, P(OEt)3,
toluene, 80 °C, 8 h; (d) 6-bromohexan-1-ol (for n = 5), 10-bromo-
decan-1-ol (for n = 9), BF3·OEt2, CH2Cl2, r.t., 24 h; (e) NaN3,
Bu4NBr, DMF, 5 h at 60 °C, and 12 h at r.t.; (f) CS2, P(OEt)3, toluene,
80 °C, 8 h.
NCS functionality at the anomeric center were prepared
following proven methods.10 Glycosides, which carry the
NCS function on an aromatic or aliphatic aglycone, re-
spectively, have also been described.11 In addition to
known NCS-functionalized sugars, which were employed
Next, thiourea-bridging with the aminohexa(ethylene gly-
col) azide [H2N(EG)6N3] was done for a series of NCS-
functionalized sugars – the glycosides 5, 10, 11, and 13 –
in this study (Table 1), long-chain alkyl a-D-mannosides,
carrying the NCS functionality at the terminus of the
aglycone portion, were targeted for the investigation of
and the well-known glycosyl isothiocyanates 12, 14, 15,
and 1610 (Table 1). The reaction was carried out at room
temperature and products were obtained after standard
workup. The respective thiourea-bridged glyco-OEG
azides were isolated in pure form after column chroma-
tography on silica gel in mostly excellent yields. Depro-
tection according to Zemplén18 led to the corresponding
unprotected derivatives 25 to 32 in nearly quantitative
yields (Table 1).
mannose-specific bacterial adhesion.12 It has been shown
earlier, that the alkyl aglycone of such mannosides can ex-
hibit high affinity to the respective bacterial lectin
FimH.13
All new glycoside isothiocyanates were synthesized from
the corresponding azides in a Staudinger-type reaction
employing carbon disulfide and triethyl phosphite.14
Thus, the 2-azidoethylmannoside 415 was converted into
the respective isothiocyanate 5 in good yield (Scheme 2).
Whereas the precursor molecule for this synthesis, man-
noside 3,16 can be obtained by glycosidation of the pen-
taacetate 1, this was not possible in case of the longer
glycosyl acceptors. 6-Bromohexan-1-ol and 10-bromo-
decan-1-ol had to be glycosylated using the mannosyl
trichloroacetimidate 2 as glycosyl donor.17 The Lewis
acid catalyzed reaction proceeded in good yields, giving
the bromo-functionalized mannosides 6 and 7, which
were in turn subjected to nucleophilic displacement with
sodium azide to yield 8 and 9, followed by conversion into
the desired isothiocyanates 10 and 11.
The carbohydrate-decorated OEG azides 25 to 32 have
been designed for 1,3-dipolar cycloaddition to alkyne-ter-
minated SAMs.To test if glyco-OEG azides are suited for
the Meldal–Sharpless ‘click’ reaction,8 the phenylmanno-
side-derived OEG azide 29 was reacted with dodec-1-yne
under copper(I) catalysis (Scheme 3). This reaction pro-
duced the 1,4-triazole 33 in very good yield after column
chromatography. The regioselectivity of the cycloaddi-
tion reaction was proved by NMR analysis.
Synthesis 2010, No. 5, 828–836 © Thieme Stuttgart · New York

830
C. Grabosch et al.
PAPER
Table 1 Synthesis of Thiourea-Bridged Glyco-OEG Azides Employing Aminohexa(ethylene glycol) Azide [H2N(EG)6N3]
NCS-functionalized carbohydrates
Thiourea-bridged glyco-OEG-azides
Yield (%)
Liga-
tiona
Depro-
tectionb
OR
OR
OAc
OAc
O
RO
RO
O
AcO
AcO
85
90
50
90
99
78
S
O
O
N3
5
17: R = Ac
25: R = H
N
H
N
H
O
O
NCS
5
5
5
OAc
OAc
OR
OR
O
O
AcO
AcO
RO
RO
S
O
O
N3
10
O
18: R = Ac
26: R = H
NCS
O
N
H
N
H
5
5
OR
OR
OAc
OAc
O
O
RO
RO
AcO
AcO
S
O
O
N3
O
11
N
H
N
H
O
19: R = Ac
27: R = H
NCS
9
9
OR
OR
OAc
OAc
O
RO
RO
O
AcO
AcO
99
68
93
H
HN
N
O
O
N3
NCS
20: R = Ac
28: R = H
12
5
S
OR
OR
OAc
OAc
O
RO
RO
O
AcO
AcO
quant
O
O
13
S
21: R = Ac
29: R = H
O
N3
N
H
N
O
NCS
H
5
OR
OAc
O
O
AcO
RO
RO
H
H
AcO
AcO
O
93
51
99
quant
N
N
N3
NCS
O
OR
5
S
22: R = Ac
30: R = H
14
OR
OAc
O
O
RO
RO
AcO
AcO
OR
OAc
99
RO
AcO
O
O
O
RO
O
AcO
H
H
O
23: R = Ac
31: R = H
N
N
NCS
N3
O
RO
5
AcO
15
S
OR
OR
OAc
OAc
O
O
O
RO
RO
O
H
H
AcO
AcO
O
O
82
O
N
N
NCS
N3
RO
O
AcO
OR
5
OR
AcO
AcO
S
24: R = Ac
32: R = H
16
a Reaction conditions: see general procedure.
b Reaction conditions: see general procedure.
Synthesis 2010, No. 5, 828–836 © Thieme Stuttgart · New York

PAPER
Glyco-SAMs by ‘Dual Click’
831
the crude product was purified by flash column chromatography on
silica gel (Table 1).
OH
OH
O
HO
HO
De-O-Acetylation; General Procedure
O
The acetylated thiourea-bridged glyco-OEG-azide 17–24 (20–226
mg) was dissolved in anhyd MeOH (10 mL) and a catalytic amount
of NaOMe was added at 0 °C. The reaction mixture was stirred at
r.t. for 3 h, followed by addition of Amberlite IR120 ion exchange
resin for neutralization. After filtration, the filtrate was concentrated
under reduced pressure to obtain the pure product (Table 1).
29
S
O
N3
N
N
O
H
H
5
a
OH
OH
86%
O
HO
HO
Me
2-Isothiocyanatoethyl 2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranoside (5)
(H2C)9
O
33
S
The acetylated azide 4(450 mg, 1.08 mmol) was dissolved in anhyd
toluene (10 mL) and CS2 (2.79 mL, 46.3 mmol, 43 equiv) and
P(OEt)3 (740 mL, 4.32 mmol, 4 equiv) were added. The mixture was
stirred for 8 h at 80 °C. Then H2O (15 mL) was added and the solu-
tion was kept at 4 °C for 12 h. After extraction with CH2Cl2 (3 × 20
mL), the organic phases were combined and dried (MgSO4). After
filtration, the filtrate was concentrated under reduced pressure and
the residual syrup purified by flash column chromatography (cyclo-
hexane–EtOAc, 2:1 → 1:2) to give the title compound as a color-
less, amorphous solid; yield: 378 mg (80%); Rf = 0.51
(cyclohexane–EtOAc, 1:2); [a]D +11 (c 1.23, CHCl3).
N
N
O
N
N
H
N
O
H
5
Scheme 3 Reagents and conditions: (a) CuI, MeCN, 70 °C, 18 h.
In conclusion, it was shown that the aminohexa(ethylene
glycol) azide [H2N(EG)6N3] can be addressed in two sub-
sequent ‘click’-type reactions to produce chain-like
biorepulsive molecules, which are needed for the con-
struction of glyco-SAMs. This concept implies a highly
flexible modular approach for fabrication of carbohy-
drate-decorated monolayers and their modification. Ac-
cording to the ‘dual click’ concept, the two
chemoselective ligation steps can be applied in any se-
quence, either in solution or ‘on SAM’. Both ligation re-
actions which were employed – thiourea-bridging and
1,3-diploar cycloaddition of azides and alkynes – are fea-
sible in carbohydrate chemistry. Owing to the chemistry
used, either method works equally well in every sugar se-
ries, which is important because carbohydrates are the
IR (ATR): 2094 (s, NCS), 1730 cm–1 (s, C=O).
3
1H NMR (600 MHz, CDCl3): d = 5.36 (dd, J2,3 = 3.4 Hz,
3
3J3,4 = 10.0 Hz, 1 H, H-3), 5.31 (t, J3,4 = 10.0 Hz, 1 H, H-4), 5.29
(dd, 3J1,2 = 1.8 Hz, 3J 2,3 = 3.4 Hz, 1 H, H-2), 4.89 (d, 3J1,2 = 1.8 Hz,
1 H, H-1), 4.30 (dd, 3J5,6 = 5.2 Hz, 3J6,6¢ = 12.4 Hz, 1 H, H-6), 4.14
(dd, 3J5,6¢ = 2.8 Hz, 3J6,6¢ = 12.4 Hz, 1 H, H-6¢), 4.09 (mc, 1 H, H-5),
3.90 (mc, 1 H, man–OCHH), 3.80–3.70 (m, 3 H, man–OCHH,
CH2NCS), 2.16, 2.11, 2.05, 2.00 (each s, each 3 H, 4 COCH3).
13C NMR (150 MHz, CDCl3): d = 170.5, 169.9, 169.8, 169.7 (4
COCH3), 134.4 (NCS), 97.9 (C-1), 69.3 (C-5), 69.1 (C-2), 68.9 (C-
3), 66.7 (man–OCH2), 66.0 (C-4), 62.5 (C-6), 45.0 (CH2NCS), 20.8,
20.7, 20.7, 20.6 (4 COCH3).
biorelevant head groups in our studies. This project will MALDI-TOF-MS: m/z [M + Na]+ calcd for C17H23NO10S + Na:
456.09; found: 456.19.
be further developed towards increasing complexity of the
sugar decoration.
6-Bromohexyl 2,3,4,6-tetra-O-acetyl-a-D-mannopyranoside (6)
To an ice-cooled solution of the mannosyl donor 2 (400 mg, 0.815
All reactions were monitored by TLC on silica gel F254 (Merck)
with detection by UV light and/or by charring with 10% ethanolic
H2SO4 and subsequent heating. Flash chromatography was per-
formed on Merck silica gel 60 (0.040–0.063 mm). NMR spectra
were recorded at 550 or 600 MHz on Bruker DRX 500 and AV 600
instruments at 298 K. Chemical shifts (d) are reported in ppm and
are relative to Me4Si (d = 0) as internal standard. IR spectra were
measured with a PerkinElmer FT-IR Paragon 1000 (ATR) spec-
trometer. ESI-MS measurements were performed on a Mariner in-
strument, MALDI-TOF mass spectra were recorded on a Bruker
Biflex III instrument with 19 kV acceleration voltage and an ioniza-
tion laser at 337 nm. As matrices 2,5-dihydroxybenzoic acid and a-
cyano-4-hydroxycinnamic acid were used. Optical rotations were
determined with a PerkinElmer 241 polarimeter at r.t. (10 cm cells,
Na D-line: 589 nm). Petroleum ether (PE) used refers to the fraction
boiling in the range 30–60 °C.
mmol) and 6-bromohexan-1-ol (107 mL, 0.976 mmol, 1.2 equiv) in
anhyd CH2Cl2 (10 mL) was added BF3·OEt2 (413 mL, 3.26 mmol,
4 equiv) slowly. After 30 min, the ice bath was removed and the
mixture was stirred r.t. overnight. Afterwards, aq NaHCO3 (40 mL)
was added and the aqueous phase was extracted with CH2Cl2
(2 × 40 mL). The combined organic phases were washed with H2O
(2 × 40 mL), dried (MgSO4), filtered, and the filtrate was concen-
trated. The product was obtained after column chromatography
(PE–EtOAc, 3:2) as a colorless syrup; yield: 320 mg (77%);
Rf = 0.43 (PE–EtOAc, 3:2); [a]D +21 (c 0.60, CH2Cl2).
IR (ATR): 2937 (s, C–H), 1740 (s, C=O), 1044 cm–1 (s, C–Br).
3
1H NMR (600 MHz, CDCl3): d = 5.35 (dd, J2,3 = 3.5 Hz,
3J3,4 = 10.0 Hz, 1 H, H-3), 5.28 (t, 3J3,4 = 10.0 Hz, 3J4,5 = 10.0 Hz, 1
3
3
H, H-4), 5.23 (d, J1,2 = 1.8 Hz, J2,3 = 3.5 Hz, 1 H, H-2), 4.80 (d,
3J1,2 = 1.8 Hz, 1 H, H-1), 4.28 (dd, 3J5,6 = 5.8 Hz, 3J6,6¢ = 12.2 Hz, 1
H, H-6), 4.12 (dd, J5,6 = 2.5 Hz, J6,6¢ = 12.2 Hz, 1 H, H-6¢), 3.98
(mc, 1 H, H-5), 3.70 (mc, 1 H, man–OCHH), 3.48–3.44 (m, 1 H,
3
3
Thiourea-Bridging; General Procedure
3
man–OCHH), 3.42 (t, J = 6.8 Hz, 2 H, CH2Br), 2.15, 2.10, 2.04,
To the O-acetylated NCS-functionalized carbohydrate 5, 10–16
(20–220 mg) dissolved in ice-cold anhyd CH2Cl2 (5 mL) were add-
ed aminohexa(ethylene glycol) azide [H2N-(EG)6-N3, 1 equiv] and
DIPEA (1.01 equiv) and the reaction mixture was stirred at 0 °C for
1 h. After removal of the ice bath, stirring was continued at r.t. over-
night. Then, the solvent was removed under reduced pressure and
2.00 (each s, each 3 H, 4 COCH3), 1.88 (mc, 2 H, CH2CH2Br), 1.88,
1.61, 1.48, 1.40 (each mc, each 2 H, 3 CH2).
13C NMR (150 MHz, CDCl3): d = 170.6, 170.1, 169.9, 169.7
(4 COCH3), 97.7 (C-1), 69.7 (C-2), 69.1 (C-3), 68.5 (C-5), 68.3
(man–OCH2), 66.4 (C-4), 62.6 (C-6), 33.7 (CH2Br), 32.6
Synthesis 2010, No. 5, 828–836 © Thieme Stuttgart · New York

832
C. Grabosch et al.
PAPER
(CH2CH2Br), 29.1 (man–OCH2CH2), 27.9 (CH2CH2CH2Br), 25.3
(man–OCH2CH2CH2), 20.9, 20.7, 20.7, 20.7 (4 COCH3).
MALDI-TOF-MS: m/z [M + Na]+ calcd for C20H31N3O10 + Na:
496.19; found: 496.28.
HRESI-MS: m/z [M + Na]+ calcd for C20H31BrO10 + Na: 533.0998;
found: 533.0993.
10-Azidodecyl 2,3,4,6-tetra-O-acetyl-a-D-mannopyranoside (9)
To a solution of the bromide 7 (660 mg, 1.16 mmol) in anhyd DMF
(10 mL) were added NaN3 (379 mg, 5.83 mmol, 5 equiv) and
Bu4NBr (74 mg, 230 mmol, 0.2 equiv). The reaction mixture was
stirred at 60 °C for 5 h and at r.t. overnight. Then, the mixture was
poured into ice water (20 mL) and extracted with CH2Cl2
(3 × 20 mL). The combined organic phases were dried (MgSO4),
filtered, and the filtrate concentrated. The residue was purified by
flash column chromatography (cyclohexane–EtOAc, 2:1) to give
the title compound as a colorless syrup; yield: 520 mg (85%);
Rf = 0.36 (cyclohexane–EtOAc, 2:1); [a]D +43 (c 0.55, CH2Cl2).
10-Bromodecyl 2,3,4,6-tetra-O-acetyl-a-D-mannopyranoside
(7)
To a solution of the trichloroacetimidate 2 (790 mg, 1.16 mmol) in
ice-cooled anhyd CH2Cl2 (20 mL) were added 10-bromodecan-1-ol
(355 mL, 1.93 mmol, 1.2 equiv) and BF3·OEt2 (410 mL, 3.22 mmol,
2 equiv). After 30 min, the ice bath was removed and the mixture
was stirred at r.t. overnight. Afterwards, aq NaHCO3 (40 mL) was
added and the aqueous phase was extracted with CH2Cl2 (2 × 40
mL). The combined organic phases were washed with H2O (20 mL),
dried (MgSO4), filtered, and the filtrate was concentrated. Column
chromatography (cyclohexane–EtOAc, 1:1) gave a colorless syrup;
yield: 690 mg (76%); Rf = 0.43 (cyclohexane–EtOAc, 1:1); [a]D
+20 (c 0.33, CH2Cl2).
IR (ATR): 2927 (s, C–H), 2094 (s, N3), 1746 cm–1 (s, C=O).
1H NMR (500 MHz, CDCl3): d = 5.28 (d, 3J2,3 = 3.4 Hz, 3J3,4 = 10.0
3
3
Hz, 1 H, H-3), 3.21 (t, J3,4 = 10.0 Hz, J4,5 = 10.0 Hz, 1 H, H-4),
3
3
5.16 (dd, J1,2 = 1.8 Hz, J2,3 = 3.4 Hz, 1 H, H-2), 4.73 (d,
IR (ATR): 2929 (s, C–H), 1746 (s, C=O), 1046 cm–1 (C–Br).
3J1,2 = 1.8 Hz, 1 H, H-1), 4.21 (dd, 3J5,6 = 5.2 Hz, 3J6,6 = 12.6 Hz, 1
3
3
H, H-6) 4.04 (dd, J5,6¢ = 2.5 Hz, J6,6¢ = 12.6 Hz, 1 H, H-6¢), 3.92
(ddd, 3J5,6¢ = 2.5 Hz, 3J5,6 = 5.2 Hz, 3J4,5 = 10.0 Hz, 1 H, H-5), 2.61
(dt, 2J = 9.6 Hz, 3J = 6.6 Hz, 1 H, man–OCHH), 3.57 (t, 3J = 6.6 Hz,
2 H, CH2N3), 3.38 (dt, 2J = 9.1 Hz, 3J = 6.6 Hz, 1 H, man–OCHH),
2.09, 2.04, 1.98, 1.93 (each s, each 3 H, 4 COCH3), 1.56–1.47 (m, 8
H, 4 CH2), 1.31–1.19 (m, 8 H, 4 CH2).
3
1H NMR (600 MHz, CDCl3): d = 5.34 (dd, J2,3 = 3.4 Hz,
3J3,4 = 10.0 Hz, 1 H, H-3), 5.26 (dd, 3J3,4 = 10.0 Hz, 3J4,5 = 10.0 Hz,
1 H, H-4), 5.22 (d, 3J1,2 = 1.4 Hz, 3J2,3 = 3.4 Hz, 1 H, H-2), 4.79 (d,
3J1,2 = 1.4 Hz, 1 H, H-1), 4.27 (dd, 3J5,6 = 5.3 Hz, 3J6,6¢ = 12.2 Hz, 1
3
3
H, H-6), 4.10 (dd, J5,6¢ = 2.4 Hz, J6,6¢ = 12.2 Hz, 1 H, H-6¢), 3.97
(ddd, 3J4,5 = 10.0 Hz, 3J5,6 = 5.3 Hz, 3J5,6¢ = 2.4 Hz, 1 H, H-5), 3.68–
3.62 (mc, 3 H, man–OCHH, man–OCH2CH2), 3.46–3.38 (m, 3 H,
man–OCHH, CH2Br), 2.15, 2.10, 2.04, 1.99 (each s, each 3 H, 4
COCH3), 1.84, 1.58 (each mc, each 4 H, 4 CH2), 1.45–1.26 (m, 6 H,
3 CH2).
13C NMR (125 MHz, CDCl3): d = 170.7, 170.2, 170.0, 169.8 (4
COCH3), 97.6 (C-1), 69.8 (C-2), 69.2 (C-3), 68.6 (man–OCH2),
68.4 (C-5), 66.3 (C-4), 62.5 (C-6), 51.5 (CH2N3), 30.0, 29.4, 29.4,
29.3, 29.2, 28.9, 26.1, 25.7 (8 CH2), 21.0, 20.8, 20.8, 20.7 (4
COCH3).
HRESI-MS: m/z [M + Na]+ calcd for C24H39N3O10 + Na: 552.2533;
found: 552.2528.
13C NMR (150 MHz, CDCl3): d = 170.7, 170.1, 169.9, 169.8
(4 COCH3), 97.6 (C-1), 69.8 (C-2), 69.2 (C-3), 68.6 (C-5), 68.4
(man–OCH2), 66.3 (C-4), 62.6 (C-6), 34.00 (CH2Br), 32.8
(CH2CH2Br), 29.5, 29.4, 29.6, 29.5, 29.3, 29.3, 28.7, 28.2 (8 CH2),
20.9, 20.7, 20.7, 20.7 (4 COCH3).
6-Isothiocyanatohexyl 2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranoside (10)
HRESI-MS: m/z [M + Na]+ calcd for C24H39BrO10 + Na: 589.1624;
To a solution of the azide 8 (363 mg, 0.767 mmol) in anhyd toluene
(10 mL) were added CS2 (1.99 mL, 40.0 mmol, 43 equiv) and
P(OEt)3 (530 mL, 3.07 mmol, 4 equiv) were added. The mixture
was stirred for 8 h at 80 °C. Then, H2O (15 mL) was added and the
solution was kept at 4 °C for 12 h. After extraction with CH2Cl2
(3 × 20 mL), the organic phases were combined, dried (MgSO4), fil-
tered, and the filtrate was concentrated. The crude product was pu-
rified by column chromatography (cyclohexane–EtOAc, 3:2) to
give the title compound as a colorless syrup; yield: 219 mg (58%);
Rf = 0.34 (cyclohexane–EtOAc, 3:2); [a]D +52 (c 0.31, CH2Cl2).
found: 589.1653.
6-Azidohexyl 2,3,4,6-tetra-O-acetyl-a-D-mannopyranoside (8)
To a solution of the bromide 6 (1.26 g, 2.47 mmol) in anhyd DMF
(10 mL) were added NaN3 (802 mg, 12.4 mmol, 5 equiv) and
Bu4NBr (159 mg, 0.494 mmol, 0.2 equiv) and the reaction mixture
was stirred at 60 °C for 5 h and overnight at r.t. Then, the mixture
was poured into ice water and extracted with CH2Cl2 (3 × 20 mL).
The combined organic phases were dried (MgSO4), filtered, and the
filtrate was concentrated. The residue was purified by flash column
chromatography (cyclohexane–EtOAc, 3:2) to obtain the title com-
pound as a colorless syrup, yield: 852 mg (73%); Rf = 0.38 (cyclo-
hexane–EtOAc, 3:2); [a]D +48 (c 0.69, CH2Cl2).
IR (ATR): 2098 (m, NCS), 1740 cm–1 (s, C=O).
3
1H NMR (500 MHz, CDCl3): d = 5.34 (dd, J3,2 = 3.4 Hz,
3J3,4 = 10.0 Hz, 1 H, H-3), 5.28 (t, 3J3,4 = 10.0 Hz, 3J4,5 = 10.0 Hz, 1
H, H-4), 5.23 (dd, 3J1,2 = 1.8 Hz, 3J2,3 = 3.4 Hz, 1 H, H-2), 4.80 (d,
3J1,2 = 1.8 Hz, 1 H, H-1), 4.28 (dd, 3J5,6 = 5.3 Hz, 3J6,6¢ = 12.2 Hz, 1
IR (ATR): 2093 (s, N3), 1741 cm–1 (s, C=O).
1H NMR (600 MHz, CDCl3): d = 5.35 (dd, J2,3 = 3.4 Hz,
3
3
3
H, H-6), 4.12 (dd, J5,6¢ = 2.5 Hz, J6,6¢ = 12.2 Hz, 1 H, H-6¢), 3.98
3J3,4 = 10.0 Hz,
1
H, H-3), 5.27 (dd~t, J3,4 = 10.0 Hz,
3
(ddd, 3J4,5 = 10.0 Hz, 3J5,6 = 5.3 Hz, 3J5,6¢ = 2.5 Hz, 1 H, H-5), 3.70
3J4,5 = 10.0 Hz, 1 H, H-4), 5.23 (d, 3J1,2 = 1.8 Hz, 3J2,3 = 3.4 Hz, 1 H,
(td, 3J = 6.4 Hz, 2J = 9.7 Hz,
1 H, man–OCHH), 3.53 (t,
3
3
H-2), 4.80 (d, J1,2 = 1.8 Hz, 1 H, H-1), 4.28 (dd, J5,6 = 5.4 Hz,
3J6,6¢ = 12.2 Hz, 1 H, H-6), 4.12 (dd, 3J5,6¢ = 2.5 Hz, 3J6,6¢ = 12.2 Hz,
1 H, H-6¢), 3.98 (ddd, 3J4,5 = 10.0 Hz, 3J5,6 = 5.4 Hz, 3J5,6¢ = 2.5 Hz,
1 H, H-5), 3.69 (mc, 1 H, man–OCHH), 3.46 (mc, 1 H, man–
3J = 6.5 Hz, 2 H, CH2NCS), 3.46 (mc, 1 H, man–OCHH), 2.16,
2.19, 2.10, 2.00 (each s, each 3 H, 4 COCH3), 1.73, 1.64 (each mc,
each 2 H, 2 CH2), 1.44 (mc, 4 H, CH2CH2).
3
OCHH), 3.28 (t, J = 6.9 Hz, 2 H, CH2N3), 2.15, 2.10, 2.04, 2.00
(each s, each 3 H, 4 COCH3), 1.63 (mc, 4 H, man–OCH2CH2,
CH2N3), 1.41 (mc, 4 H, CH2CH2CH2CH2N3).
13C NMR (150 MHz, CDCl3): d = 170.6, 170.1, 169.9, 169.7
(4 COCH3), 97.6 (C-1), 69.8 (C-2), 69.2 (C-3), 68.5 (C-5),
68.3 (man–OCH2), 66.4 (C-4), 62.6 (C-6), 51.4 (CH2N3),
13C NMR (125 MHz, CDCl3): d = 170.6, 170.1, 169.9, 169.7 (4
COCH3), 105.7 (NCS), 97.6 (C-1), 69.7 (C-2), 69.1 (C-3), 68.5 (C-
5), 68.2 (man–OCH2), 66.3 (C-4), 62.6 (C-6), 45.0 (CH2NCS), 29.8,
29.1 (2 CH2CH2CH2), 26.4, 25.4 (CH2CH2CH2CH2CH2CH2NCS),
20.9, 20.7, 20.7, 20.7 (4 COCH3).
HRESI-MS: m/z [M + Na]+ calcd for C21H31NO10S + Na: 512.1566;
29.2,
28.8
(CH2CH2CH2CH2CH2CH2N3), 26.5, 25.7
found: 512.1555.
(CH2CH2CH2CH2CH2CH2N3), 20.9, 20.7, 20.7, 20.7 (4 COCH3).
Synthesis 2010, No. 5, 828–836 © Thieme Stuttgart · New York

PAPER
Glyco-SAMs by ‘Dual Click’
833
10-Isothiocyanatodecyl 2,3,4,6-tetra-O-acetyl-a-D-manno-
pyranoside (11)
IR (ATR): 2359 (s, N–H), 2103 (m, N3), 1743 (s, C=O), 1080 cm–1
(s, C=S).
To a solution of the azide 9 (500 mg, 0.944 mmol) in anhyd toluene
(10 mL) were added CS2 (2.45 mL, 40.6 mmol, 43 equiv) and
P(OEt)3 (658 mL, 3.78 mmol, 4 equiv). The mixture was stirred for
8 h at 80 °C. Then H2O (15 mL) was added and the solution was
kept at 4 °C for 12 h. After extraction with CH2Cl2 (3 × 20 mL), the
organic phases were combined and dried (MgSO4), filtered, and the
filtrate was concentrated. The pure product was obtained after col-
umn chromatography (cyclohexane–EtOAc, 3:2) as a colorless syr-
up; yield: 286 mg (56%); Rf = 0.30 (cyclohexane–EtOAc, 3:2); [a]D
+31 (c 0.49, CH2Cl2).
1H NMR (500 MHz, CDCl3): d = 6.60 (br s, 1 H, NH), 6.51 (br s, 1
H, NH), 5.34 (dd, 3J2,3 = 3.4 Hz, 3J3,4 = 10.0 Hz, 1 H, H-3), 5.28 (t,
3J = 10.0 Hz, 1 H, H-4), 5.23 (dd, 3J1,2 = 1.8 Hz, 3J2,3 = 3.4 Hz, 1 H,
H-2), 4.79 (d, J1,2 = 1.8 Hz, 1 H, H-1), 4.28 (dd, J5,6 = 5.3 Hz,
3J6,6¢ = 12.2 Hz, 1 H, H-6), 4.11 (dd, 3J5,6¢ = 2.5 Hz, 3J6,6¢ = 12.2 Hz,
1 H, H-6¢), 3.98 (ddd, 3J4,5 = 10.0 Hz, 3J5,6 = 2.5 Hz, 3J6,6¢ = 5.2 Hz,
1 H, H-5), 3.73–3.68 (m, 28 H, man–OCHH, HHCNHCS,
CSNHCH2, 6 CH2OCH2), 3.47–3.43 (m, 2 H, man–OCHH, HHCN-
HCS), 3.39 (t, 3J = 5.1 Hz, 2 H, CH2N3), 2.16, 2.10, 2.05, 1.99 (each
s, each 3 H, 4 COCH3), 1.61 (mc, 4 H, 2 CH2CH2CH2), 1.39 (mc, 4
H, man–OCH2CH2CH2CH2CH2CH2NHCS).
3
3
IR (ATR): 2927 (s, C–H), 2093 (s, NCS), 1744 cm–1 (s, C=O).
1H NMR (500 MHz, CDCl3): d = 5.28 (d, 3J2,3 = 3.4 Hz, 3J3,4 = 10.0
13C NMR (125 MHz, CDCl3): d = 182.8 (C=S), 170.6, 170.1,
169.9, 169.7 (4 COCH3), 97.5 (C-1), 70.7–70.0 (6 CH2OCH2), 69.7
(C-2), 69.2 (C-3), 68.6 (C-5), 68.4 (man–OCH2), 66.3 (C-4), 62.5
(C-6), 50.7 (CH2N3), 44.7 (H2CNHCS), 29.1, 29.0 (2 CH2CH2CH2),
26.7, 25.9 (man–OCH2CH2CH2CH2CH2CH2NHCS), 20.9, 20.7,
20.7, 20.7 (4 COCH3).
3
3
Hz, 1 H, H-3), 3.21 (t, J3,4 = 10.0 Hz, J4,5 = 10.0 Hz, 1 H, H-4),
3
3
5.17 (dd, J1,2 = 1.7 Hz, J2,3 = 3.4 Hz, 1 H, H-2), 4.73 (d,
3J1,2 = 1.7 Hz, 1 H, H-1), 4.05 (dd, 3J5,6 = 5.3 Hz, 3J6,6 = 12.6 Hz, 1
3
3
H, H-6) 4.04 (dd, J5,6¢ = 2.4 Hz, J6,6¢ = 12.6 Hz, 1 H, H-6¢), 3.2
(ddd, 3J5,6¢ = 2.4 Hz, 3J5,6 = 5.3 Hz, 3J4,5 = 10.0 Hz, 1 H, H-5), 2.60
(dt, 2J = 9.6 Hz, 3J = 6.6 Hz, 1 H, man–OCHH), 3.44 (t, 3J = 6.6 Hz,
2 H, CH2NCS), 3.38 (dt, 2J = 9.1 Hz, 3J = 6.6 Hz, 1 H, man–
OCHH), 2.09, 2.04, 1.98, 1.93 (each s, each 3 H, 4 COCH3), 1.63,
1.53, 1.35 (each m, each 2 H, 3 CH2), 1.30–1.21 (m, 10 H, 5 CH2).
MALDI-TOF-MS: m/z [M + Na]+ calcd for C35H61N5O16S + Na:
862.37; found: 862.22.
N-[10-(2,3,4,6-Tetra-O-acetyl-a-D-mannopyranosyloxy)do-
decyl]-N¢-[w-azidohexa(ethylene glycol)] Thiourea (19)
The acetylated isothiocyanate 11 (200 mg, 0.367 mmol) was treated
according to the general procedure. After purification by column
chromatography (EtOAc–MeOH; 30:1), the product was obtained
as a colorless oil; yield: 165 mg (50%); Rf = 0.43 (EtOAc–MeOH,
30:1).
13C NMR (125 MHz, CDCl3): d = 170.7, 170.2, 170.0, 169.8 (4
C=O), 163.5 (NCS), 97.6 (C-1), 69.8 (C-2), 69.2 (C-3), 68.6 (man–
OCH2), 68.4 (C-5), 66.3 (C-4), 62.5 (C-6), 45.1 (CH2NCS), 29.9,
29.4, 29.3, 29.3, 29.2, 28.8, 26.5, 26.1 (8 CH2), 20.9, 20.8, 20.7,
20.7 (4 COCH3).
HRESI-MS: m/z [M + Na]+ calcd for C25H39NO10S + Na: 552.2192;
found: 568.2187.
3
1H NMR (600 MHz, CDCl3): d = 5.28 (dd, J2,3 = 3.5 Hz,
3J3,4 = 10.0 Hz, 1 H, H-3), 5.21 (dd, 3J3,4 = 10.0 Hz, 3J4,5 = 10.0 Hz,
1 H, H-4), 5.16 (dd, 3J1,2 = 1.8 Hz, 3J2,3 = 3.5 Hz, 1 H, H-2), 4.73 (d,
3J1,2 = 1.8 Hz, 1 H, H-1), 4.22 (dd, 3J5,6 = 5.3 Hz, 3J6,6¢ = 12.2 Hz, 1
N-[2-(2,3,4,6-Tetra-O-acetyl-a-D-mannopyranosyloxy)ethyl]-
N¢-[w-azidohexa(ethylene glycol)] Thiourea (17)
The acetylated isothiocyanate 5 (115 mg, 0.266 mmol) was thio-
urea-bridged according to the general procedure. Column chroma-
tography (EtOAc–MeOH, 30:1) gave the pure product as a colorless
oil; yield: 141 mg (68%); Rf = 0.41 (EtOAc–MeOH, 30:1); [a]D +23
(c 1.37, CH2Cl2).
3
3
H, H-6), 4.13 (dd, J5,6 = 2.4 Hz, J6,6¢ = 12.2 Hz, 1 H, H-6¢), 3.99
(ddd, 3J5,6¢ = 2.4 Hz 3J5,6 = 5.3 Hz, 3J4,5 = 10.0 Hz, 1 H, H-5), 3.63–
3.53 (m, 28 H, man–OCHH, HHCNHCS, SCNHCH2, 6 CH2OCH2),
3.37 (mc, 2 H, HHCNHCS, 1 H, man–OCHH), 3.32 (t, 3J = 5.1 Hz,
2 H, CH2N3), 2.09, 2.04, 1.98, 1.93 (each s, each 3 H, 4 COCH3),
1.56–1.47 (m, 4 H, man–OCH2CH2, SCNHCH2CH2), 1.30–1.18 (m,
12 H, 6 CH2).
IR (ATR): 2358 (s, N–H), 2102 (s, N3), 1742 (s, C=O), 1078 cm–1
(s, C=S).
13C NMR (150 MHz, CDCl3): d = 182.5 (C=S), 170.7, 170.1, 169.9,
169.8 (4 COCH3), 97.6 (H-1), 70.7–70.0 (6 CH2OCH2), 69.7 (C-2),
69.2 (C-3), 68.6 (man–OCH2), 68.4 (C-5), 66.3 (C-4), 62.5 (C-6),
50.7 (CH2N3), 44.8 (CH2NHCS), 29.5, 29.5, 29.4, 29.3, 29.2, 27.5,
27.0, 26.1 (8 CH2), 20.9, 20.8, 20.8, 20.7 (4 COCH3).
HRESI-MS: m/z [M + Na]+ calcd for C39H69N5O16S + Na:
918.4358; found: 918.4352.
1H NMR (600 MHz, CDCl3): d = 7.00 (br s, 1 H, NH), 6.88 (br s, 1
H, NH), 5.29 (mc, 1 H, H-3), 5.26 (mc, 1 H, H-4), 5.24 (dd,
3J1,2 = 1.5 Hz, 3J2,3 = 3.1 Hz, 1 H, H-2), 4.84 (d, 3J1,2 = 1.5 Hz, 1 H,
H-1), 4.27 (dd, 3J5,6 = 5.4 Hz, 3J6,6¢ = 12.2 Hz, 1 H, H-6), 4.13 (dd,
3J5,6 = 2.4 Hz, 3J6,6¢ = 12.2 Hz, 1 H, H-6¢), 3.99 (mc, 1 H, H-5), 3.95
(mc, 1 H, HHCNHCS), 3.90 (mc, 1 H, man–OCHH), 3.74–3.60 (m,
28 H, man–OCHH, HHCNHCS, SCNHCH2, 6 CH2OCH2), 3.39 (t,
3J = 5.1 Hz, 2 H, CH2N3), 2.15, 2.11, 2.05 1.98 (each s, each 3 H, 4
COCH3).
13C NMR (150 MHz, CDCl3): d = 183.5 (C=S), 170.7, 170.0,
169.9, 169.6 (4 COCH3), 97.6 (H-1), 70.7–70.0 (CH2O), 69.4 (C-2),
69.2 (C-3), 68.7 (C-5), 67.2 (man–OCH2), 66.2 (C-4), 62.5 (C-6),
50.7 (CH2N3), 44.7, 44.0 (CH2NHCSNHCH2), 20.8, 20.8, 20.7,
20.6 (4 COCH3).
N-(2,3,4,6-Tetra-O-acetyl-a-D-mannopyranosyl)-N¢-[w-azido-
hexa(ethylene glycol)] Thiourea (20)
The acetylated isothiocyanate 12 (216 mg, 0.557 mmol) was thio-
urea-bridged according to the general procedure. Column chroma-
tography (EtOAc–MeOH, 30:1) gave the pure product as a colorless
oil; yield: 352 mg (85%); Rf = 0.40 (EtOAc–MeOH, 30:1).
IR (ATR): 2359 (s, N–H), 2104 (s, N3), 1742 (s, C=O), 1048.7
cm–1 (s, C=S).
MALDI-TOF-MS: m/z [M + Na]+ calcd for C31H53N5O16S + Na:
806.31; found: 807.10.
1H NMR (500 MHz, CDCl3): d = 7.42 (br s, 1 H, NH), 7.32 (br s, 1
N-[6-(2,3,4,6-Tetra-O-acetyl-a-D-mannopyranosyloxy)hexyl]-
N¢-[w-azidohexa(ethylene glycol)] Thiourea (18)
The isothiocyanate 10 (109 mg, 0.222 mmol) was thiourea-bridged
as described in the general procedure. After column chromatogra-
phy (EtOAc–MeOH, 30:1), the product was obtained as a colorless
oil; yield: 117 mg (63%); Rf = 0.38 (EtOAc–MeOH, 30:1); [a]D +23
(c 0.52, CH2Cl2).
H, NH), 5.84 (br s, 1 H, H-1), 5.30–5.25 (m, 3 H, H-2, H-3, H-4),
3
4.33 (dd, J5,6 = 5.2 Hz, 3J6,6¢ = 12.2 Hz, 1 H, H-6), 4.13 (dd,
3J5,6 = 2.3 Hz, 3J6,6¢ = 12.2 Hz, 1 H, H-6¢), 4.02 (m, 1 H, H-5), 3.82
(m, 2 H, SCNHCH2), 3.73–3.62 (m, 24 H, 6 CH2OCH2), 3.39 (t,
3J = 5.1 Hz, 2 H, CH2N3), 2.16, 2.09, 2.05, 2.02 (each s, each 3 H, 4
COCH3).
Synthesis 2010, No. 5, 828–836 © Thieme Stuttgart · New York

834
C. Grabosch et al.
PAPER
13C NMR (125 MHz, CDCl3): d = 183.3 (C=S), 170.7, 169.9, 169.9,
169.5 (4 COCH3), 80.3 (C-1), 70.7–70.5 (6 CH2OCH2) 70.0 (C-5),
69.1 (C-3), 68.8 (C-2), 66.4 (C-4), 62.2 (C-6), 50.7 (CH2N3), 45.2
(SCNHCH2), 20.8, 20.8, 20.7, 20.7 (4 COCH3).
HRESI-MS: m/z [M + Na]+ calcd for C29H49N5O15S + Na:
762.2844; found: 762.2869.
(t, 3J3,4 = 9.9 Hz, 3J4,5 = 9.9 Hz, 1 H, H-4¢), 4.81–4.77 (m, 2 H, H-2,
H-2¢), 4.36 (dd, 3J5,6 = 2.3 Hz, 3J6,6 = 12.1 Hz, 1 H, H-6a), 4.19 (dd,
3J5,6¢ = 4.2 Hz, 3J6,6¢ = 12.1 Hz, 1 H, H-6b), 4.14 (dd, 3J5,6¢ = 3.6 Hz,
3J6,6¢ = 12.5 Hz, 1 H, H-6a¢), 3.98 (dd, 3J5,6 = 2.3 Hz, 3J6,6 = 12.5 Hz,
3
3
1 H, H-6b¢), 3.93 (t, J3,4 = 9.3 Hz, J4,5 = 9.3 Hz, 1 H, H-4), 3.85
3
3
(mc, 1 H, H-5), 3.77 (ddd, J4,5 = 9.4 Hz, 3J5,6 = 2.3 Hz, J5,6¢ = 3.6
Hz, 1 H, H-5¢), 3.74–3.48 (m, 26 H, 6 CH2OCH2, CSNHCH2), 3.31
(t, 3J = 5.0 Hz, 2 H, CH2N3), 2.05, 2.03, 2.00, 2.96, 2.04, 1.93, 1.92
(each s, each 3 H, 7 COCH3).
N-[4-(2,3,4,6-Tetra-O-acetyl-a-D-mannopyranosyloxy)phenyl]-
N¢-[w-azidohexa(ethylene glycol)] Thiourea (21)
The NCS-functionalized mannoside 13 (120 mg, 250 mmol) was
thiourea-bridged according to the general procedure. After purifica-
tion by column chromatography (EtOAc–MeOH, 30:1), the product
was obtained as a colorless oil; yield: 206 mg (99%); Rf = 0.39
(EtOAc–MeOH, 30:1); [a]D +47 (c 0.62, CH2Cl2).
13C NMR (150 MHz, CDCl3): d = 184.1 (C=S), 170.7, 170.5,
170.5, 170.4, 169.8, 169.8, 169.7, 169.5 (7 COCH3), 95.5 (C-1¢),
82.0 (C-1), 76.0 (C-3¢), 73.5 (C-5¢), 72.7 (C-4), 71.3 (C-2, C-2¢),
70.6–70.0 (6 CH2OCH2, glc–OCH2), 69.3 (C-3), 68.4 (C-5), 67.9
(C-4¢), 62.9 (C-6), 61.7 (C-6¢), 50.6 (CH2N3), 20.8, 20.8, 20.7, 20.7,
20.6, 20.6, 20.6 (7 COCH3).
IR (ATR): 2358 (s, N–H), 2103 (s, N3), 1744 (s, C=O), 1506 (m,
C=C), 1085 cm–1 (m, C=S).
MALDI-TOF-MS: m/z [M + Na]+ calcd for C41H65N5O23S + Na:
1050.36; found: 1050.92.
1H NMR (600 MHz, CDCl3): d = 7.98 (br s, 1 H, NH), 7.29 (d,
3
3J = 8.8 Hz, 2 Harom, Hx, Hx¢), 7.10 (d, J = 8.8 Hz, 2 Harom, Hy,
N-(2,2¢,3,3¢,4¢,6,6¢-Hepta-O-acetyl-b-D-cellobiosyl)-N¢-[w-azido-
hexa(ethylene glycol)] Thiourea (24)
The acetylated isothiocyanate 16 (22.6 mg, 34.0 mmol) was thio-
urea-bridged according to the general procedure. After column
chromatography (EtOAc–MeOH, 5:1), the compound was obtained
as a colorless, amorphous solid; yield: 37 mg (99%); Rf = 0.58
(EtOAc–MeOH, 5:1).
Hy¢), 6.70 (br s, 1 H, NH), 5.54 (dd, 3J2,3 = 3.5 Hz, 3J3,4 = 10.0 Hz,
1 H, H-3), 5.49 (d, 3J1,2 = 1.8 Hz, 1 H, H-1), 5.43 (dd, 3J1,2 = 1.8 Hz,
3J2,3 = 3.5 Hz, 1 H, 2-H), 5.38 (dd~t, 3J3,4 = 10.0 Hz,3J4,5 = 10.0 Hz,
1 H, H-4), 4.29 (mc, 1 H, H-6), 4.10 (mc, 2 H, H-5, H-6¢), 3.83 (br s,
2 H, CSNHCH2), 3.60–3.58 (m, 24 H, 6 CH2OCH2), 3.37 (t, J = 5.2
Hz, 2 H, CH2N3), 2.20, 2.06, 2.05, 2.04 (each s, each 3 H,
4 COCH3).
13C NMR (150 MHz, CDCl3): d = 181.5 (C=S), 170.5, 169.9,
169.9, 169.7 (4 COCH3), 153.9 (man–OCar), 142.8 (Car–NH), 126.6
(aryl-Cx), 117.4 (aryl-Cy), 96.1 (C-1), 70.7–70.0 (6 CH2OCH2),
69.3 (C-5), 69.3 (C-2), 68.8 (C-3), 65.9 (C-4), 62.1 (C-6), 50.7
(CH2N3), 44.9 (SCNHCH2), 21.0, 20.8, 20.7, 20.7 (4 COCH3).
1H NMR (600 MHz, CDCl3): d = 7.51 (br s, 1 H, NH), 7.10 (br s, 1
H, NH), 5.77 (br s, 1 H, H-1), 5.26 (t, 3J2,3 = 8.6 Hz, 3J3,4 = 8.6 Hz,
1 H, H-3), 5.12 (t, 3J2,3 = 9.4 Hz, 3J3,4 = 9.4 Hz, 1 H, H-3¢), 5.06 (t,
3J3,4 = 9.4 Hz, 3J4,5 = 9.4 Hz, 1 H, H-4¢), 4.92 (mc, 2 H, H-2, H-2¢),
3
4.49 (d, J1,2 = 7.9 Hz, 1 H, H-1¢), 4.46 (mc, 1 H, H-6a), 4.35 (dd,
3J5,6 = 4.2 Hz, 3J6,6 = 12.4 Hz, 1 H, H-6b), 4.13 (dd, 3J5,6¢ = 4.2 Hz,
HRESI-MS: m/z [M + Na]+ calcd for C35H53N5O16S + Na:
854.3106; found: 854.3162.
3J6,6¢ = 12.1 Hz,
1
H, H-6a¢), 4.20 (dd, 3J5,6¢ = 2.0 Hz,
3J6,6¢ = 12.4 Hz, 1 H, H-6b¢), 3.79–3.55 (m, 29 H, H-4, H-5, H-5¢, 6
3
CH2OCH2, CSNHCH2), 3.37 (t, J = 5.0 Hz, 2 H, CH2N3), 2.09,
N-(2,3,4,6-Tetra-O-acetyl-b-D-glucopyranosyl)-N¢-[w-azido-
hexa(ethylene glycol)] Thiourea (22)
Glycosyl isothiocyanate 14 (125 mg, 0.321 mmol) was thiourea-
bridged as described in the general procedure. Column chromatog-
raphy (EtOAc–MeOH, 30:1) gave the pure product as a colorless
oil; yield: 213 mg (90%); Rf = 0.35 (EtOAc–MeOH, 30:1).
2.08, 2.02, 2.01, 2.00, 1.99, 1.97 (each s, each 3 H, 7 COCH3).
13C NMR (150 MHz, CDCl3): d = 184.1 (C=S), 170.7, 170.5,
170.2, 170.2, 169.5, 169.3, 169.0 (7 COCH3), 100.5 (C-1), 82.2 (C-
1¢), 76.4 (C-4), 74.1 (C-5), 72.9 (C-3, C-3¢), 71.9 (C-5¢), 71.5 (C-2,
C-2¢), 70.8–69.8 (6 CH2OCH2), 67.8 (C-4¢), 62.0 (C-6), 61.6 (C-6¢),
50.6 (CH2N3), 20.9, 20.8, 20.7, 20.5 (7 COCH3).
MALDI-TOF-MS: m/z [M + Na]+ calcd for C41H65N5O23S + Na:
1050.36; found: 1050.78.
1H NMR (500 MHz, CDCl3): d = 7.47 (br s, 1 H, NH), 7.23 (br s, 1
3
H, NH), 5.89 (br s, 1 H, H-1), 5.32 (dd~t, J2,3 = 9.4 Hz,
3J3,4 = 9.4 Hz, 1 H, H-3), 5.07 (dd~t, J = 9.5 Hz, 1 H, H-4), 5.01
3
3
(dd~t, J1,2 = 9.4 Hz, 3J2,3 = 9.4 Hz, 1 H, H-2), 4.31 (dd, J5,6 = 4.4
N-[2-(a-D-Mannopyranosyloxy)ethyl]-N¢-[w-azidohexa(ethyl-
ene glycol)] Thiourea (25)
The acetylated mannoside 17 (80 mg, 102 mmol) was deprotected
according to the general procedure to give a colorless oil; yield:
56 mg (90%); [a]D +24 (c 0.75, MeOH).
IR (ATR): 3324 (br, O–H), 2918 (s, C–H), 2106 (s, N3), 1087 cm–1
(s, C=S).
Hz, 3J6,6¢ = 12.4 Hz,
3J6,6¢ = 12.4 Hz,
1
H, H-6), 4.09 (dd, 3J5,6¢ = 2.2 Hz,
H, H-6¢), 3.84 (ddd, J4,5 = 10.1 Hz,
3
1
3J5,6 = 4.4 Hz, 3J5,6¢ = 2.2 Hz, 1 H, H-5), 3.73–3.59 (m, 26 H,
CSNHCH2, 6 CH2OCH2), 3.39 (t, J = 5.2 Hz, 2 H, CH2N3), 2.06,
2.03, 2.03, 2.00 (each s, each 3 H, 4 COCH3).
13C NMR (125 MHz, CDCl3): d = 184.2 (C=S), 170.8, 169.8,
169.9, 169.6 (4 COCH3), 82.5 (C-1), 73.5 (C-3), 73.2 (C-5), 70.1
(C-2), 70.7–70.0 (6 CH2OCH2), 69.9 (NHCH2), 68.4 (C-4), 61.8
(C-6), 50.7 (CH2N3), 20.7, 20.7, 20.7, 20.5 (4 COCH3).
MALDI-TOF-MS: m/z [M + Na]+ calcd for C29H49N5O15S + Na:
762.28; found: 763.45.
1H NMR (600 MHz, CD3OD): d = 4.82 (d, 3J1,2 = 1.6 Hz, 1 H, H-1),
3.89 (mc, 1 H, H-6), 3.87 (dd, 3J1,2 = 1.6 Hz, 3J2,3 = 3.1 Hz, 1 H, H-
2), 3.76–3.66 (m, 33 H, H-3, H-4, H-6¢, man–OCH2, man–
3
OCH2CH2, 6 CH2OCCH2, CSNHCH2), 3.58 (ddd, J4,5 = 9.8 Hz,
3
3
3J5,6 = 2.3 Hz, J5,6¢ = 5.9 Hz, 1 H, H-5), 3.42 (t, J = 5.1 Hz, 2 H,
CH2N3).
N-(2,2¢,3,3¢,4¢,6,6¢-Hepta-O-acetyl-b-D-maltosyl)-N¢-[w-azido-
hexa(ethylene glycol)] Thiourea (23)
13C NMR (150 MHz, CD3OD): d = 182.8 (C=S), 101.7 (C-1), 74.7
(C-5), 72.5 (C-3), 72.0 (C-2), 71.6–71.1 (6 CH2OCH2), 68.5 (C-4),
67.3 (man–OCH2), 62.87 (C-6), 51.8 (CH2N3), 45.3, 45.2
(CH2NHCS, SCNHCH2).
HRESI-MS: m/z [M + Na]+ calcd for C23H45N5O12S + Na:
638.2678; found: 638.2701.
The acetylated disaccharide 15 (100 mg, 148 mmol) was thiourea-
bridged according to the general procedure. Column chromatogra-
phy (cyclohexane–EtOAc, 1:5) gave the pure product as a colorless
oil; yield: 77 mg (51%); Rf = 0.15 (cyclohexane–EtOAc, 1:5).
1H NMR (600 MHz, CDCl3): d = 7.32 (br, 1 H, NH), 7.10 (br, 1 H,
NH), 5.82 (br, 1 H, H-1), 5.34–5.27 (m, 3 H, H-1¢, H-3, H-3¢), 4.99
Synthesis 2010, No. 5, 828–836 © Thieme Stuttgart · New York

PAPER
Glyco-SAMs by ‘Dual Click’
835
N-[6-(a-D-Mannopyranosyloxy)hexyl]-N¢-[w-azidohexa(ethyl-
ene glycol)] Thiourea (26)
The acetylated mannoside 18 (226 mg, 269 mmol) was deprotected
according to the general procedure. A colorless oil was obtained;
yield: 178 mg (99%); [a]D +29 (c 0.52, MeOH).
N-[4-(a-D-Mannopyranosyloxy)phenyl]-N¢-[w-azidohexa(ethyl-
ene glycol)] Thiourea (29)
Mannoside 21 (300 mg, 361 mmol) was deprotected according to
the general procedure. A colorless oil was obtained; yield: 240 mg
(quant); [a]D +76 (c 0.89, MeOH).
IR (ATR): 3314 (br, O–H), 2922 (s, C–H), 2104 (s, N3), 1552 (s, N–
H), 1077 cm–1 (s, C=S).
1H NMR (600 MHz, CDCl3): d = 7.17 (d, 3J = 8.6 Hz, 2 Harom, Hx,
Hx¢), 7.98 (d, 3J = 8.6 Hz, 2 Harom, Hy, Hy¢), 5.46 (br s, 1 H, H-1),
4.06 (br, 1 H, H-2), 3.96 (dd, 3J2,3 = 2.6 Hz, 3J3,4 = 9.9 Hz, 1 H, H-
3), 3.88 (mc, 1 H, H-6), 3.76 (mc, 2 H, NHCH2), 3.66–3.55 (m, 27
H, H-4, H-5, H-6¢, 6 CH2OCH2), 3.36 (t, 3J = 5.1 Hz, 2 H, CH2N3).
13C NMR (150 MHz, CDCl3): d = 181.2 (C=S), 154.5 (man–OCar),
132.3 (Car–NH), 126.6 (aryl-Cx), 117.2 (aryl-Cy), 98.5 (C-1), 73.2
(C-5), 71.3 (C-3), 70.6–70.0 (6 CH2OCH2), 70.2 (C-2), 69.5 (C-4),
61.1 (C-6), 50.7 (CH2N3), 45.0 (SCNHCH2).
1H NMR (600 MHz, CD3OD): d = 4.74 (d, 3J1,2 = 1.7 Hz, 1 H, H-1),
3
3.82 (dd, J5,6 = 2.4 Hz, 3J6,6¢ = 11.8Hz, 1 H, H-6), 3.79 (dd,
3
3
3J1,2 = 1.7 Hz, J2,3 = 3.4 Hz, 1 H, H-2), 3.71 (dd, J5,6¢ = 5.9 Hz,
3J6,6¢ = 11.8 Hz, 1 H, H-6¢), 3.69–3.64 (m, 29 H, 6 CH2OCH2,
SCNHCH2, man–OCH2, H-3), 3.61 (t, 3J = 9.6 Hz, 2 H, CH2NHCS)
3
3
3
3.51 (ddd, J4,5 = 9.5 Hz, J5,6 = 2.0 Hz, J5,6¢ = 6.0 Hz, 1 H, H-5),
3
3
3
3.51 (dd, J3,4 = 6.2 Hz, J4,5 = 9.5 Hz, 1 H, H-4), 3.40 (t, J = 5.0
Hz, 2 H, CH2N3), 1.66–1.56 (m, 4 H, CH2CH2CH2CH2), 1.46–1.36
(mc, 4 H, man–OCH2CH2CH2CH2CH2CH2NHCS).
HRESI-MS: m/z [M + Na]+ calcd for C27H45N5O12S + Na:
686.2683; found: 686.2678.
13C NMR (150 MHz, CD3OD): d = 170.4 (C=S), 101.6 (C-1), 75.1
(C-5), 73.1 (C-3), 72.7 (C-2), 72.0–71.2 (6 CH2OCH2), 69.1 (C-4),
68.9 (man–OCH2), 63.4 (C-6), 52.2 (CH2N3), 44.6, 45.5
(CH2NHCS, SCNHCH2), 31.0, 28.2, 27.6 (4 CH2).
HRESI-MS: m/z [M + Na]+ calcd for C27H53N5O12S + Na:
694.3304; found: 694.3299.
N-(b-D-Glucopyranosyl)-N¢-[w-azidohexa(ethylene glycol)]
Thiourea (30)
Thiourea derivative 22 (37 mg, 0.186 mmol) was deprotected as de-
scribed in the general procedure to obtain the title compound as col-
orless oil; yield: quant; [a]D –3 (c 0.81, MeOH).
IR (ATR): 3325 (br, O–H), 2341 (s, N–H), 2100 (s, N3), 1073 cm–1
N-[10-(a-D-Mannopyranosyloxy)decyl]-N¢-[w-azidohexa(ethyl-
ene glycol)] Thiourea (27)
The acetylated mannoside 19 (165 mg, 184 mmol) was deprotected
according to the general procedure to give a colorless oil; yield:
105 mg (78%); [a]D +32 (c 0.57, MeOH).
(s, C=S).
1H NMR (500 MHz, CD3OD): d = 5.29 (br s, 1 H, H-1), 3.89 (dd,
3J5,6 = 2.3 Hz, 3J6,6¢ = 12.0 Hz, 1 H, H-6), 3.81 (br s, 2 H, NHCH2),
3.73–3.67 (m, 25 H, H-6¢, 6 CH2OCH2), 3.48–3.40 (m, 4 H, H-3, H-
5, CH2N3), 3.36–3.31 (m, 2 H, H-2, H-4).
IR (ATR): 3330 (br, O–H), 2922 (s, C–H), 2102 (s, N3), 1550 (s, N–
H), 1083 cm–1 (C=S).
13C NMR (125 MHz, CD3OD): d = 185.5 (C=S), 85.0 (C-1), 79.2
(C-5), 78.9 (C-3), 74.2 (C-2), 71.0 (C-4), 71.5–70.3 (6 CH2OCH2),
62.8 (C-6), 51.8 (CH2N3), 45.5 (SCNHCH2).
HRESI-MS: m/z [M + Na]+ calcd for C21H41N5O11S + Na:
594.2421; found: 594.2416.
1H NMR (500 MHz, CD3OD): d = 4.73 (d, 3J1,2 = 1.6 Hz, 1 H, H-1),
3
3
3.82 (dd, J5,6 = 2.4 Hz, J6,6 = 11.7 Hz1 H,, H-6), 3.78 (dd,
3J1,2 = 1.6 Hz, 3J2,3 = 3.3 Hz, 1 H, H-2), 3.66–3.27 (m, 31 H, H-3, H-
4, H-6¢, man–OCH2, 6 CH2OCH2, CHHNHCS, SCNHCHH), 3.52
3
(ddd, 3J4,5 = 9.4 Hz, J5,6 = 2.4 Hz, 3J5,6¢ = 5.7 Hz, 1 H, H-5), 3.44–
N-[b-D-Maltosyl]-N¢-[w-azidohexa(ethylene glycol)] Thiourea
(31)
The acetylated disaccharide 23 (72 mg, 0.068 mmol) was deprotect-
ed according to the general procedure to give a colorless oil; yield:
49 mg (99%); [a]D +37 (c 0.10, MeOH).
3
3.40 (m, 2 H, CHHNHCS, SCNHCHH), 3.38 (t, J = 4.9 Hz, 2 H,
CH2N3), 1.52–1.44 (m, 4 H, 2 CH2), 1.30–1.24 (m, 12 H, 6 CH2).
13C NMR (125 MHz, CD3OD): d = 183.7 (C=S), 102.0 (C-1), 75.0
(C-2), 73.1, 72.7 (C-3, C-5), 72.1–71.2 (6 CH2OCH2), 69.1 (C-4),
69.0 (man–OCH2), 63.4 (C-6), 52.2 (CH2N3), 45.7, 45.5
(CH2NHCS, SCNHCH2), 31.1, 31.1, 31.0, 30.7, 28.4, 27.8 (8 CH2).
HRESI-MS: m/z [M + Na]+ calcd for C31H61N5O12S + Na:
750.3930; found: 750.3969.
IR (ATR): 3314 (br, O–H), 2872 (s, C–H), 2104 (s, N3), 1549 (s,
N3), 1072 cm–1 (s, C=S).
1H NMR (600 MHz, CD3OD): d = 5.22 (d, 3J1,2 = 3.8 Hz, 1 H, H-1),
3
3.91 (dd, J5,6 = 1.9 Hz, 3J6,6¢ = 12.3 Hz, 1 H, H-6a), 3.89 (dd,
3J5,6 = 1.8 Hz, 3J6,6¢ = 12.4 Hz, 1 H, H-6a¢), 3.84 (dd, 3J5,6 = 4.5 Hz,
3J6,6¢ = 12.3 Hz, 1 H, H-6b), 3.82–3.78 (m, 2 H, SCNHCH2), 3.75–
3.59 (m, 29 H, 6 CH2OCH2, H-3, H-3¢, H-2¢, H-5¢, H-6b¢), 3.59 (t,
3J3,4 = 9.3 Hz, 3J4,5 = 9.3 Hz, 1 H, H-4), 3.51–3.49 (m, 2 H, H-2, H-
N-(a-D-Mannopyranosyl)-N¢-[w-azidohexa(ethylene glycol)]
Thiourea (28)
The acetylated derivative 20 (116 mg, 0.157 mmol) was deprotect-
ed according to the general procedure to give a colorless oil; yield:
83.0 mg (93%); [a]D +4 (c 1.45, MeOH).
IR (ATR): 3297 (br, O–H), 2359 (s, N–H), 2094 (N3), 1069 cm–1
(C=S).
1H NMR (600 MHz, CD3OD): d = 5.65 (br s, 1 H, H-1), 3.89 (dd,
3J5,6 = 2.3 Hz, 3J6,6¢ = 11.8 Hz, 1 H, H-6), 3.86 (br s, 1 H, H-2), 3.76
(mc, 2 H, NHCH2), 3.73–3.63 (m, 25 H, H-6¢, 6 CH2OCH2), 3.55
(mc, 2 H, H-3, H-4), 3.40 (t, 3J = 5.0 Hz, 2 H, CH2N3), 3.32 (m, 1 H,
H-5).
3
5), 3.42 (t, 3J = 5.0 Hz, 2 H, CH2N3), 3.32 (t, J3,4 = 9.3 Hz,
3J4,5 = 9.3 Hz, 1 H, H-4¢).
13C NMR (150 MHz, CD3OD): d = 185.6 (C=S), 102.8 (C-1), 81.0
(C-4), 78.7 (C-5¢), 77.8 (C-5), 77.7, 74.7 (C-3, C-3¢), 74.1 (C-2),
73.8 (C-2¢), 71.6–70.2 (6 CH2OCH2), 71.1 (C-4¢), 63.0 (C-6), 62.2
(C-6¢) 51.8 (CH2N3), 45.6 (SCNHCH2).
HRESI-MS: m/z [M + Na]+ calcd for C27H51N5O16S + Na:
756.2944; found: 756.2944.
13C NMR (150 MHz, CD3OD): d = 184.6 (C=S), 83.2 (C-1), 79.6
(C-5), 75.8 (C-3), 72.4 (C-2), 71.6–70.8 (6 CH2OCH2), 68.3 (C-4),
63.0 (C-6), 51.8 (CH2N3), 45.7 (SCNHCH2).
HRESI-MS: m/z [M + Na]+ calcd for C21H41N5O11S + Na:
594.2421; found: 594.2416.
N-(b-D-Cellobiosyl)-N¢-[w-azidohexa(ethyleneglycol)]Thiourea
(32)
The acetylated disaccharide 24 (37 mg, 0.033 mmol) was deprotect-
ed according to the general procedure to give a colorless oil; yield:
19 mg (82%); [a]D +36 (c 0.31, MeOH).
IR (ATR): 3312 (br, O–H), 2920 (s, C–H), 2108 (s, N3), 1549 (w,
N–H), 1073 cm–1 (s, C=S).
Synthesis 2010, No. 5, 828–836 © Thieme Stuttgart · New York

836
C. Grabosch et al.
PAPER
M.; Röckendorf, N.; Lindhorst, T. K. Eur. J. Org. Chem.
2004, 3931. (d) Ban, L.; Mrksich, M. Angew. Chem. Int. Ed.
2008, 47, 3396; Angew. Chem. 2008, 120, 3444.
(5) (a) Kind, M.; Wöll, C. Prog. Surf. Sci. 2009, 84, 230.
(b) Liang, P.-H.; Wu, C.-Y.; Greenberg, W. A.; Wong, C.-H.
Curr. Opin. Chem. Biol. 2008, 12, 86.
1H NMR (600 MHz, CD3OD): d = 4.46 (d, 3J1,2 = 7.8 Hz, 1 H, H-1),
3
3.95–9.91 (m, 2 H, H-6a, H-6a¢), 3.89 (dd, J5,6 = 3.9 Hz,
3J6,6¢ = 12.0 Hz, 1 H, H-6b), 3.73–3.60 (m, 31 H, 6 CH2OCH2,
SCNHCH2, H-6b¢, H-5¢, H-4, H-4¢, H-3), 3.42 (m~t, 2 H, CH2N3),
3
3.40–3.37 (m,
2
H, H-5, H-3¢), 3.28 (dd, J1,2 = 7.8 Hz,
3J2,3 = 9.2 Hz, 1 H, H-2).
(6) (a) Prime, K. L.; Whitesides, G. M. Science 1991, 252,
1164. (b) Herrwerth, S.; Eck, W.; Reinhardt, S.; Grunze, M.
J. Am. Chem. Soc. 2003, 125, 9359. (c) Chelmowski, R.;
Köster, S. D.; Kerstan, A.; Prekelt, A.; Grunwald, C.;
Winkler, T.; Metzler-Nolte, N.; Terfort, A.; Wöll, C. J. Am.
Chem. Soc. 2008, 130, 14952.
13C NMR (150 MHz, CD3OD): d = 185.7 (C = S), 108.3 (C-1¢),
80.6 (C-5¢), 78.2 (C-3¢), 78.1, 77.9, 77.9, 77.7, 77.4 (C-2, C-3, C-4,
C-4¢, C-5), 74.9 (C-2¢), 71.62–71.1 (6 CH2OCH2, SCNHCH2), 62.5
(C-6), 61.9 (C-6), 51.8 (CH2N3).
MALDI-TOF-MS: m/z [M + Na]+ calcd for C27H51N5O16S + Na:
756.29; found: 756.71.
(7) Kleinert, M.; Winkler, T.; Terfort, A.; Lindhorst, T. K. Org.
Biomol. Chem. 2008, 6, 2118.
N-[4-(a-D-Mannopyranosyloxy)phenyl]-N¢-[4-decyl-1H-1,2,3-
triazol-1-ylhexa(ethylene glycolyl)] Thiourea (33)
(8) (a) Tornøe, C. W.; Christensen, C.; Meldal, M. J. Org.
Chem. 2002, 67, 3057. (b) Rostovtsev, V. V.; Green, L. G.;
Fokin, V. V.; Sharpless, K. B. Angew. Chem. Int. Ed. 2002,
41, 2596; Angew. Chem. 2002, 114, 2708.
(9) (a) Lindhorst, T. K.; Kieburg, C. Angew. Chem., Int. Ed.
Engl. 1996, 35, 1953; Angew. Chem. 1996, 108, 2083.
(b) Ortiz Mellet, C.; Defaye, J.; García-Fernández, J. M.
Chem. Eur. J. 2002, 8, 1983. (c) Köhn, M.; Benito, J. M.;
Ortiz Mellet, C.; Lindhorst, T. K.; García Fernández, J. M.
ChemBioChem 2004, 5, 771.
Azide 29 (38 mg, 58 mmol) and dodec-1-yne (25.0 mL, 116 mmol)
were dissolved in degassed MeCN (10 mL). After flushing with N2,
CuI (2.2 mg, 12 mmol, 0.2 equiv) was added and the reaction mix-
ture warmed up to 70 °C for 18 h. After removing the solvent, the
residue was purified by flash column chromatography (CH2Cl2–
MeOH, 5:1). The triazole 33 was obtained as light yellow oil; yield:
40 mg (86%); Rf = 0.52 (CH2Cl2–MeOH, 5:1).
1H NMR (500 MHz, CD3OD): d = 7.77 (s, 1 H, HC=), 7.26 (d,
3
3J = 8.8 Hz, 2 Harom, Hx, Hx¢), 7.13 (d, J = 8.8 Hz, 2 Harom, Hy,
(10) (a) Lindhorst, T. K.; Kieburg, C. Synthesis 1995, 1228.
(b) Kühne, M.; Györgydeák, Z.; Lindhorst, T. K. Synthesis
2006, 949.
3
3
Hy¢), 5.46 (d, J1,2 = 1.8 Hz, 1 H, H-1), 4.53 (t, J = 4.8 Hz, 2 H,
SCNHCH2), 4.01 (dd, J1,2 = 1.8 Hz, 3J2,3¢ = 3.4 Hz, 1 H, H-2),
3
(11) (a) Walter, M.; Lindhorst, T. K. Monatsh. Chem. 2002, 133,
473. (b) Free, P.; Hurley, C. A.; Kageyama, T.; Chain, B.
M.; Tabor, A. B. Org. Biomol. Chem. 2006, 4, 1817.
(12) (a) Ohlsen, K.; Oelschlaeger, T. A.; Hacker, J.; Khan, A. S.
Top. Curr. Chem. 2009, 288, 109. (b) Klemm, P.;
Schembri, M. Int. J. Med. Microbiol. 2000, 290, 27.
(c) Mulvey, M. A. Cell. Microbiol. 2002, 4, 257.
(d) Dubber, M.; Sperling, O.; Lindhorst, T. K. Org. Biomol.
Chem. 2006, 4, 3901. (e) Bouckaert, J.; Mackenzie, J.;
de Paz, J. L.; Chipwaza, B.; Choudhury, D.; Zavialov, A.;
Mannerstedt, K.; Anderson, J.; Pierard, D.; Wyns, L.;
Seeberger, P. H.; Oscarson, S.; De Greve, H.; Knight, S. D.
Mol. Microbiol. 2006, 61, 1556.
3.90–3.86 (m, 3 H, H-3, SCNHCH2CH2), 3.78–3.69 (m, 5 H, H-4,
H-6, H-6¢, CH2–Ctriazole), 3.66–3.57 (m, 23 H, H-5, OCH2, 5
3
CH2OCH2), 2.68 (t, J = 7.6 Hz, 2 H, Ctriazole–CH2), 1.66 (m, 2 H,
Ctriazole–CH2CH2), 1.34–1.29 (m, 12 H, 6 CH2), 0.89 (t, 3J = 7.0 Hz,
3 H, CH3).
13C NMR (125 MHz, CD3OD): d = 182.5 (C=S), 156.0 (man–
OCar), 149.1 (Car–NH), 133.8 (Ctriazole), 127.6 (aryl-Cx), 124.0
(CHtriazole), 118.5 (aryl-Cy), 100.4 (C-1), 75.5 (C-5), 72.4 (C-3), 71.9
(C-2), 71.6–70.5 (6 CH2OCH2), 68.3 (C-4), 62.7 (C-6), 51.3
(SCNHCH2), 45.5 (CH2Ctriazole), 33.1 (Ctriazole–CH2CH2), 30.7, 30.7,
30.6, 30.5, 30.5, 30.3 (6 CH2CH2CH2), 26.3 (Ctriazole–CH2), 23.8
(CH2), 17.5 (CH2CH3).
MALDI-TOF-MS: m/z [M + Na]+ calcd for C39H67N5O13S + Na:
836.46; found: 836.82.
(13) Bouckaert, J.; Berglund, J.; Schembri, M.; De Genst, E.;
Cools, L.; Wuhrer, M.; Hung, C.-S.; Pinkner, J.; Slättegård,
R.; Zavialov, A.; Choudhury, D.; Langermann, S.; Hultgren,
S. J.; Wyns, L.; Klemm, P.; Oscarson, S.; Knight, S. D.;
De Greve, H. Mol. Microbiol. 2005, 55, 441.
Acknowledgment
(14) García-Moreno, M. I.; Díaz-Pérez, P.; Benito, J. M.;
Ortiz Mellet, C.; Defaye, J.; Garcia Fernádez, J. M.
Carbohydr. Res. 2002, 337, 2329.
We thank Prof. Dr. A. Terfort (Frankfurt a. M.) for fruitful discus-
sions and an excellent cooperation about glyco-SAMs.
(15) Cheryak, A. Y.; Sharma, G. V.; Kononov, L. O.; Krishna, P.
R.; Levinsky, A. B.; Kochetkov, N. K.; Rao, A. V. R.
Carbohydr. Res. 1992, 223, 303.
(16) (a) Hasegawa, T.; Fujisawa, T.; Numata, M.; Matsumoto, T.;
Umeda, M.; Karinaga, R.; Mizu, M.; Koumoto, K.; Kimura,
T.; Okumura, S.; Sakurai, K.; Shinkai, S. Org. Biomol.
Chem. 2004, 2, 3091. (b) Dahmen, J.; Frjd, T.; Groenberg,
G.; Lave, T.; Magnusson, G.; Noori, G. Carbohydr. Res.
1983, 118, 292.
(17) (a) Jung, K.-H.; Hoch, M.; Schmidt, R. R. Liebigs Ann.
Chem. 1989, 1099. (b) Schmidt, R. R.; Kinzy, W. Adv.
Carbohydr. Chem. Biochem. 1994, 50, 21. (c) Lindhorst, T.
K.; Kötter, S.; Krallmann-Wenzel, U.; Ehlers, S. J. Chem.
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Synthesis 2010, No. 5, 828–836 © Thieme Stuttgart · New York