Synthesis and biological evaluation of S-neofucopeptides as E- and P-selectin inhibitors

Article abstract of DOI:10.1002/ejoc.200800199

The synthesis of α/β-L-fucosylated cysteamine, 3-thiopropionic acid, and 3-thioacetic acid derivatives as building blocks for the preparation of S-neofucopeptides is shown. These compounds were used in the synthesis of new thiofucosides derivatives (8α, 9α, 9β, 10α, 22α, 22β, 24α, 26α) that show affinity towards E- and P-selectins. They constitute a new series of hydrolytically stable and low-molecular-weight mimetics of the natural SLe^x tetrasaccharide. Wiley-VCH Verlag GmbH & Co. KGaA, 2008.

Full text of DOI:10.1002/ejoc.200800199

FULL PAPER
DOI: 10.1002/ejoc.200800199
Synthesis and Biological Evaluation of S-Neofucopeptides as E- and P-Selectin
Inhibitors
Antonio J. Moreno-Vargas,*[a] Lidia Molina,[a] Ana T. Carmona,[a] Alessandro Ferrali,[a]
Martine Lambelet,[b] Olivier Spertini,[b] and Inmaculada Robina*[a]
Keywords: Carbohydrates / Peptides / Inhibitors / Glycoconjugates
The synthesis of α/β-
L
-fucosylated cysteamine, 3-thiopro-
show affinity towards E- and P-selectins. They constitute a
new series of hydrolytically stable and low-molecular-weight
mimetics of the natural SLex tetrasaccharide.
(© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim,
Germany, 2008)
pionic acid, and 3-thioacetic acid derivatives as building
blocks for the preparation of S-neofucopeptides is shown.
These compounds were used in the synthesis of new thiofu-
cosides derivatives (8α, 9α, 9β, 10α, 22α, 22β, 24α, 26α) that
Introduction
The carbohydrate–protein recognition between tetrasac-
charide sialyl Lewis X (SLex) (Figure 1) and E- or P-selectin
mediates the early stage of an inflammatory response,
which leads to the migration of leukocytes from the blood
stream to inflamed tissue.[1] It has been envisaged that dis-
ruption of these selectin–carbohydrate interactions might
serve as a novel target for palliative treatment of acute and
chronic inflammatory diseases.[2] SLex is a leading structure
in the design and development of selectin antagonists, and
this tetrasaccharide was identified as an antiinflammatory
agent, although its binding affinity is relatively low.[3] The
complex saccharidic structure of SLex and its sensitivity
towards acid and enzymatic hydrolysis, which makes it
orally inactive and unstable in the blood stream,[4] has pro-
moted the search for compounds that could act as SLex but
possess a simpler structure, higher binding affinity, and be
resistant to enzymatic hydrolysis.[5–8]
The common design element is based on the nature of
the known pharmacophores,[9] which are the hydroxy
groups of -fucose and -galactose, and the carboxyl group
of the sialic acid, which are all involved in the recognition
between SLex and E-selectin. The GlcNAc moiety merely
acts as a scaffold to guarantee the effective conformation
for binding.[10] Although non-carbohydrate structures have
been examined,[6] the majority of these approaches have fo-
Figure 1. Structure of the natural tetrasaccharide sialyl Lewis X
(SLex).
cused on carbohydrate-type ligands[7] and related structures
consisting of hybrid molecules[8,11,12] with varying degrees
of similarity to SLex. O-Fucopeptides incorporating resi-
dues of threonine and hydroxyproline amino acids instead
of GlcNAc and -galactose and glutaric acid instead of
sialic acid have been described.[11,12] Although these com-
pounds have shown good affinity towards E- and P-selec-
tins, the difficult glycosylation of N-acylated β-hydroxy
amino acid derivatives, caused by the decreased nucleophi-
licity of the glycosyl acceptors coming from the hydrogen-
bonding interaction between the acceptor OH and NH
groups[13] and the lability of the O-glycosidic linkage repre-
sent drawbacks in the case of their use as drugs. C-Glyco-
side[14] mimetics as non-hydrolizable mimetics of SLex as
well as S-linked analogues of carbohydrate antigens of SLex
have been prepared and characterized;[15] however, their
thioglycopeptide counterparts are relatively unexplored.[16]
We now present the synthesis of new S-linked fucosides
I–III (Figure 2) containing the functional groups deemed
critical for binding to selectins. The introduction of a sulfur
group at the anomeric position provides good acid- and en-
zyme-mediated hydrolytic stability.[16] In addition, the
higher water solubility of sulfur derivatives relative to that
of their oxygen counterparts is an important advantage for
[a] Department of Organic Chemistry, Faculty of Chemistry, Uni-
versity of Seville,
P. O. Box 553, 41071 Sevilla, Spain
Fax: +34954624960
E-mail: ajmoreno@us.es
robina@us.es
[b] Service and Central Laboratory of Hematology, Centre Hopit-
alier Universitaire Vaudoise de Lausanne,
BH 18-544, 1011 Lausanne, Switzerland
Supporting information for this article is available on the
WWW under http://www.eurjoc.org or from the author.
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A. J. Moreno-Vargas, I. Robina et al.
FULL PAPER
our compounds with regard to tolerance by most biological
systems.[17] Compounds of type I incorporate glutaric acid
instead of neuraminic acid and hydroxyproline derivatives
instead of -galactose as in the O-fucopeptides designed by
Wong.[11,12] Compounds II and III bear a trihydroxyalkyl-
furan aminoester moiety as a novel non-proteinogenic poly-
hydroxylated amino acid, which could be able to mimic
both the galactose and the sialic acid residues of SLex. Ad-
ditionally, the furan ring could favor affinity to selectins by
aromatic–aromatic interactions. The overall synthetic pro-
cedure of the target compounds is highly advantageous due
to the superior nucleophilicity of sulfur, which makes the
fucosylation reaction proceed readily, and to the use of sim-
pler spacers[18] derived from cysteamine (compounds I) or
from 3-thiopropionic/thioacetic acid (compounds II and
III).
Scheme 1. Synthesis of the S-linked fucoside building blocks.
block 4α from thiofucoside 1α, probably due to the con-
sumption of the highly reactive alkylating agent tert-butyl
bromoacetate by the excess amount of diethylamine. So, a
simple “two-step, one-pot” reaction was required to carry
out S-fucosylation of the bromoacetate derivative. Thus,
reaction of thiofucoside 1α with NaOMe in anhydrous
methanol followed by acidic resin neutralization gave 1-
mercaptofucose, which was subjected to nucleophilic substi-
tution with tert-butyl bromoacetate in the presence of Et3N
Figure 2. Structure of the new S-linked fucosides.
Results and Discussion
Synthesis
Compounds I–III were prepared from α/β-amino- to furnish the desired S-linked fucosyl derivative. This com-
alkylthiofucopyranosides 2α and 5β or from α/β-carboxy- pound was peracetylated to afford 4α in 61% overall yield.
alkylthiofucopyranosides 3α, 3β, and 4α. Two main strate- The anomeric configuration was also preserved under these
gies were used for the preparation of compounds 2–5 and conditions. Compound 3β was obtained in a similar way in
they include (a) nucleophilic displacement of the appropri- 68% yield starting from compound 1β. Alternatively, the
ate bromo derivative by an anomeric glycosyl thiolate nu- synthesis of N-Fmoc-aminoethyl β-fucoside building block
cleophile and (b) glycosylation of a thiol-containing com- 5β[21] was carried out in good yield by glycosylation of N-
pound such as cysteamine (Scheme 1).
Fmoc cysteamine with tetra-O-acetyl fucose as a donor in
Condensation of -fucose with thioacetic acid in the the presence of TMSOTf. Compounds 2–5 are attractive
presence of hydrogen chloride followed by acetylation in and versatile building blocks for the generation of libraries
pyridine gave 1-thio-α- and -β--fucose tetraacetate 1α of new S-neofucopeptides.[22]
(45%) and 1β (20%) according to the procedure of Hashim-
The synthesis of thiofucoside derivatives of 4-hy-
oto[19] et al. (Scheme 1). One-pot reaction of thioacetate 1α droxyproline (compounds of type I) was carried out in a
with N-Boc-bromoethylamine or tert-butyl 3-bromopropi- linear sequence by using standard peptide chemistry
onate with the use of diethylamine in DMF[20] gave the S- (Scheme 2).
linked-fucoside building blocks 2α and 3α in good yield.
N-Deprotection of compounds 2α or 5β followed by
Under such reaction conditions, selective deacetylation at coupling with commercial N-Fmoc-4--hydroxyproline
the sulfur atom takes place while retaining the anomeric by using (benzotriazol-1-yloxy)tripyrrolidinophosphonium
integrity. This procedure failed in the synthesis of building hexafluorophosphate/diisopropyl(ethyl)amine
(PyBOP/
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S-Neofucopeptides as E- and P-Selectin Inhibitors
starting from -glucose. Regioselective methoxytritylation
of the readily available tetrahydroxyalkyl furan 12[26] af-
forded 13 in 81% yield (Scheme 3). Reaction of 13 with
thionyl chloride and triethylamine afforded a mixture of cy-
clic sulfites, which after reaction with trimethylsilyl azide
in the presence of TBAF by using THF as solvent gave
azidoderivative 16 in 76% yield (two steps) as the unique
product. The cyclic sulfite opening occurred through an
SN2 mechanism, which allowed the total stereoselective in-
troduction of the azide functionality. After removal of the
4-methoxytrityl group in 16, followed by hydrogenation of
the azide group under atmospheric pressure, desired amino-
trihydroxyalkylfuryl ester 18 was obtained in nearly quanti-
tative yield. In order to confirm the C-1 configuration in
18, azido derivative 17 was treated sequentially with TsCl/
Py and H2/Pd for chemical correlation with the known α-
-fucosidase inhibitor 20[27] (Scheme 4). This reaction se-
quence demonstrates the success of this methodology in the
synthesis of imino-C-heterocycles.
Scheme 2. Synthesis of S-linked fucosides containing 4-hydroxy-
proline.
DIEA) as coupling reagents gave 6α or 6β in 67–85% over-
all yield. Fmoc-group removal (20% diethylamine in DMF)
and condensation with mono-tert-butyl glutaric acid gave
adduct 7α or 7β in 56–63% yield. Hydrolysis of the acetyl
groups with tBuOH/Et3N/H2O (2:1:1) afforded cleanly the
S-linked fucoside 8α or 8β. These mild deacetylation condi-
tions do not need neutralization with ion-exchange resins
as normally happens when Zemplén methodology is em-
ployed. The compounds so-obtained are free of polyanions
(traces) released from the resins (bleeding), which are diffi-
cult to detect by routine analysis and that can also mask
selectin binding assays.[23] Finally, deprotection of the tert-
butyl ester by using TFA (20%)/CH2Cl2 afforded acid 9α
or 9β in quantitative yield. Amide 10α was prepared from
7α by acidic deprotection followed by condensation with
butylamine and deacetylation.
The synthesis of tetritol-1-yl-furyl derivatives of type II
and III was carried out by starting from carboxyalkylthio-
fucosides 3α, 3β, and 4α by coupling with synthetic 4Ј-ami-
notrihydroxyalkylfuryl ester 11 and 1Ј-aminotrihydroxyalk-
ylfuryl ester 18. Compound 11 can be easily obtained from
-glucose in four steps as was described previously by our
research group.[24] We also recently reported a methodology
for the stereoselective synthesis of α-furfurylamines starting
from -aldopentoses.[25] As an extension of this work, we
Scheme 3. Stereoselective synthesis of the new 1Ј-aminotrihydroxy-
alkylfuryl ester 18.
Removal of the tert-butyl ester group in compounds 3α,
now describe the preparation of ethyl 5-(1-amino-1-deoxy- 3β, and 4α with TFA (20%)/CH2Cl2 followed by coupling
-arabino-tetritol-1-yl)-2-methylfuran-3-carboxylate (18) with aminopolyol derivatives 11 and 18 by using PyBOP/
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A. J. Moreno-Vargas, I. Robina et al.
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Table 1. IC50 [m] values for the affinity of compounds 8, 9, 10,
22, 24, 26, and SLex sodium salt toward E- and P-selectins by an
ELISA test.
Head 1[a]
8α
9α
9β 10α 22α 22β 24α 26α SLex
E-Selectin 2.5 3.5 3.4 3.1 2.4 5.7 3.0 2.8
0.7
P-Selectin 2.3 3.0[a] 3.2 2.4 2.2 5.1 2.6 2.5 n.i.[b]
Scheme 4. Synthesis of imino-C-heterocycle 20.
[a] In the presence of CaCl2 (1 m), the value obtained is 2.5 m.
[b] No inhibition under these assay conditions at 8 m.
DIEA as condensing reagents afforded adducts 21α, 21β,
23α, and 25α in moderate-to-good yields. Deacetylation
with EtOH/Et3N/H2O (2:1:1) afforded cleanly the S-linked
fucosides 22α, 22β, 24α, and 26α, which could be isolated
in 55–76% yield after final purification (Scheme 5).
We have observed that α-thiofucoside derivatives of
furylamino polyols (22α, 24α, 26α) are more active than the
β-analogue 22β, as expected considering that the fucose
moiety is α-linked in the natural tetrasaccharide. However,
α- and β-thiofucoside derivatives of 4-hydroxyprolines
(compounds 9α and 9β) present similar inhibition. The best
inhibitory values are obtained for compounds 8α and 22α,
which show IC50 = 2.5 and 2.4 m, respectively, towards E-
selectin, that is, 3.5-fold weaker than SLex but, remarkably,
an IC50 = 2.3 and 2.2 m, respectively, towards P-selectin
under conditions where SLex shows no inhibition. Non-
acidic compounds 8α and 10α with tert-butyl ester and bu-
tylamide moieties, respectively, show slightly better inhibi-
tory values than compound 9α with a free carboxylic acid
group. Simpler neutral analogues 24α and 26α having a tri-
hydroxyalkylfuran ester moiety are also active. These results
could be in agreement with the 3D structure of the E-selec-
tin/SLex complex determined by Somers et al.,[29] and the
quantum chemical calculations on the SLex molecule re-
ported by Pichierri et al.[30] These compounds may be con-
sidered a type of uncharged selectin inhibitors of which
there are few examples in the literature.[31,14c]
Scheme 5. Synthesis of S-linked fucosides containing aminotri-
hydroxyalkylfuryl esters.
Conclusions
We showed the synthesis of α/β--fucosylated cysteamine
derivatives (2α, 5β), α/β--fucosylated 3-thiopropionic acid
derivatives (3α, 3β), and α--fucosylated 3-thioacetic acid
derivative (4α) as building blocks for the preparation of S-
Biological Evaluation of Thiofucoside Derivatives Towards
E- and P-Selectins
The biological activity of ester derivatives 8α, 22α, 22β, neofucopeptides. These compounds were used in the syn-
24α, and 26α, acid derivatives 9α and 9β, amide derivative thesis of new thiofucoside derivatives (8α, 9α, 9b, 10α, 22α,
10α, and tetrasaccharide SLex itself evaluated in an ELISA- 22b, 24α, 26α) that show affinity towards E- and P-selectins;
based assay[28] was determined as their IC50 values in in- therefore, they act as non-hydrolyzable mimetics of the nat-
hibiting the SLex–BSA binding to E-selectin and the PSGL- ural SLex tetrasaccharide. Most of them are neutral com-
1 binding to P-selectin (Table 1). All the new SLex mimetics pounds that additionally incorporate not only polar func-
presented here are recognized by E- and P-selectins and tionalities but also lipophilic moieties. This fact together
show inhibitory activities in the low m range; under our with their good hydrolytic stability and their easy prepara-
assay conditions, tetrasaccharide SLex has an IC50 = 0.7 m tion will provide new possibilities for the discovery of low-
against E-selectin and it is inactive against P-selectin (at molecular-weight blockers of E- and P-selectin that are dif-
8 m concentration). We must be aware that because of the ferent from the conventional molecules incorporating ionic
absence of a universal assay to measure selectin binding in- groups. In view of these interesting results, further modifi-
teractions[5,7b,14a] and because of the difficulty in reproduc- cations of our compounds have to be continued in order to
ing the same assay results, the IC50 values so-obtained for find better candidates that could lead to a new generation
the new compounds should not be directly compared to of selectin antagonists in the µ or n range. Progress in
those of other reported SLex analogues.
this area is currently underway in our laboratory.
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S-Neofucopeptides as E- and P-Selectin Inhibitors
petroleum ether, 1:5) and was obtained in 68% yield as a colorless
Experimental Section
oil. [α]2D5 = 7.4 (c = 0.95, CH2Cl2). 1H NMR (300 MHz, CDCl3,
General Methods: Optical rotations were measured in a 1.0-cm or
1.0-dm tube with a Perkin–Elmer 241MC spectropolarimeter. 1H
and 13C NMR spectra were obtained for solutions in CDCl3, [D6]-
DMSO, CD3OD, and D2O. All the assignments were confirmed by
2D NMR experiments. The FAB mass spectra were obtained by
using glycerol or 3-nitrobenzyl alcohol as the matrix. TLC was per-
formed on silica gel HF254 (Merck), detection by UV light, and
charring with H2SO4 or with Pancaldi reagent [(NH4)6MoO4,
Ce(SO4)2, H2SO4, H2O]. Silica gel 60 (Merck, 230 mesh) was used
for preparative chromatography. All reagents and solvents were
purchased from Sigma–Aldrich. Tetrasaccharide sialyl Lewis X
(SLex) sodium salt was purchased from Calbiochem.
25 °C): δ = 5.21 (d, J3,4 = 2.8 Hz, 1 H, 4-H), 5.14 (t, J1,2 = J2,3
=
9.9 Hz, 1 H, 2-H), 4.97 (dd, J2,3 = 9.9 Hz, J3,4 = 2.8 Hz, 1 H, 3-
H), 4.43 (d, J1,2 = 9.9 Hz, 1 H, 1-H), 3.75 (q, J5,CH = 6.4 Hz, 1 H
5-H), 2.82 (m, 1 H, 1Јa-H, 1Јb-H), 2.50 (t, J2,1Јa = J2,1Јb = 7.4 Hz,
2 H, 2Јa-H, 2Јb-H), 2.11 (s, 3 H, CH3CO-), 1.98 (s, 3 H,
CH3CO-), 1.91 (s, 3 H, CH3CO-), 1.38 [s, 9 H, (CH3)3C-], 1.15 (d,
J5,CH = 6.4 Hz, 3 H, CH3 of fucose) ppm. 13C NMR (75 MHz,
CDCl3, 25 °C): δ = 171.0, 170.7, 170.1, 169.7 (4 -COO-), 83.7 (1-
C), 80.9 [(CH3)C-], 73.2 (4-C), 72.3 (3-C), 70.4 (2-C), 67.3 (5-C),
36.3 (2Ј-C), 28.1 [(CH3)3C-], 25.3 (1Ј-C), 20.8, 20.7, 20.6 (3
CH3CO-), 16.4 (CH3 of fucose) ppm. MS (FAB): m/z = 457 [M +
Na]+. HRMS (FAB): calcd. for C19H30O9SNa [M + Na]+ 457.1508;
found 457.1522.
2-(tert-Butoxycarbonylamino)ethyl
2,3,4-Tri-O-acetyl-1-thio-α-L-
fucopyranoside (2α): To a solution of thiofucoside 1α[19] (379 mg,
1.09 mmol) and N-Boc-2-bromoethylamine (235 mg, 1.05 mmol) in
dry DMF (6 mL) was added Et2NH (0.7 mL). The mixture was
stirred at room temperature for 1 h and then evaporated. Purifica-
tion of the residue by flash column chromatography (AcOEt/petro-
leum ether, 1:2) afforded 2α (400 mg, 82%) as a colorless oil. [α]2D0
(tert-Butoxycarbonyl)methyl 2,3,4-Tri-O-acetyl-1-thio-α-L-fucopyr-
anoside (4α): To a solution of thiofucoside 1α (1.06 g, 3.05 mmol)
in dry MeOH (20 mL) was added MeONa/MeOH (1 ) dropwise
until basic pH. After stirring for 3 h at room temperature, the mix-
ture was neutralized with IR-120 H+ resin, filtered, and concen-
trated. The resulting crude was redissolved in dry MeOH (20 mL)
and cooled to 0 °C. Triethylamine (0.94 mL, 6.71 mmol) and tert-
butyl bromoacetate (1.26 mL, 8.23 mmol) were added, and the mix-
ture was stirred at room temperature for 2 h. The solvent was then
evaporated, and the residue was conventionally acetylated (15 mL
of Ac2O/15 mL of Py) overnight. Evaporation and chromato-
graphic purification (AcOEt/petroleum ether, 1:4) afforded 4α
(781 mg, 61%) as a white solid. [α]2D0 = –193 (c = 1.04, CH2Cl2).
= –174 (c = 0.95, CH Cl ). IR: ν = 3419, 2983, 2095, 1747, 1643,
˜
2
2
1
1517, 1369, 1226, 1059 cm–1. H NMR (300 MHz, CDCl3, 25 °C):
δ = 5.67 (d, J1,2 = 5.2 Hz, 1 H, 1-H), 5.28 (br. d, J3,4 = 3.3 Hz, 1
H, 4-H), 5.20 (dd, J2,3 = 10.9 Hz, J1,2 = 5.2 Hz, 1 H, 2-H), 5.18
(dd, J2,3 = 10.9 Hz, J3,4 = 3.3 Hz, 1 H, 3-H), 4.90 (br. s, 1 H,
NHBoc), 4.45 (q, J5,CH = 6.5 Hz, 1 H, 5-H), 3.33–3.26 (m, 2 H,
2Јa-H, 2Јb-H), 2.79–2.60 (m, 2 H, 1Јa-H, 1Јb-H), 2.16 (s, 3 H,
CH3CO), 2.07 (s, 3 H, CH3CO), 1.99 (s, 3 H, CH3CO), 1.44 {s, 9
H, [(CH3)3C-]}, 1.16 (d, J5,CH = 6.5 Hz, 3 H, CH3 of fucose) ppm.
13C NMR (75 MHz, CDCl3, 25 °C): δ = 170.5, 170.2, 169.9 (3-
COO-), 155.6 (-CO- of Boc), 82.8 (1-C), 79.5 [(CH3)C-], 70.8 (4-
C), 68.5 (3-C), 67.9 (2-C), 65.0 (5-C), 40.2 (2Ј-C), 30.9 (1Ј-C), 28.4
[(CH3)3C-], 20.8, 20.6, 20.5 (3 CH3CO-), 15.9 (CH3 of fucose) ppm.
MS (FAB): m/z = 472 [M + Na]+. HRMS (FAB): calcd. for
C19H31NO9SNa [M + Na]+ 472.1617; found 472.1627.
IR: ν = 2980, 1748, 1370, 1221, 1137, 1083, 1061, 967, 916 cm–1.
˜
1H NMR (300 MHz, CDCl3, 25 °C): δ = 5.79 (d, J1,2 = 5.4 Hz, 1
H, 1-H), 5.29 (m, 2 H, 2-H, 4-H), 5.22 (dd, J2,3 = 10.9 Hz, J3,4
=
3.0 Hz, 1 H, 3-H), 4.47 (br. q, J5,CH = 6.5 Hz, 1 H, 5-H), 3.27 (d,
J1Јa,1Јb = 15.0 Hz, 1 H, 1Јa-H), 3.07 (d, J1Јa,1Јb = 15.0 Hz, 1 H, 1Јb-
H), 2.16 (s, 3 H, CH3CO-), 2.07 (s, 3 H, CH3CO-), 1.99 (s, 3 H,
CH3CO-), 1.46 {s, 9 H, [(CH3)3C-]}, 1.15 (d, J5,CH = 6.5 Hz, 3 H,
CH3 of fucose) ppm. 13C NMR (75 MHz, CDCl3, 25 °C): δ =
170.6, 170.2, 170.0, 169.0 (4 -COO-), 82.4 (1-C), 82.0 [(CH3)C-],
71.1 (4-C), 68.7 (3-C), 68.0 (2-C), 65.4 (5-C), 32.5 (1Ј-C), 28.1
[(CH3)3C-], 20.9, 20.8, 20.7 (3 CH3CO-), 16.1 (CH3 of fucose) ppm.
MS (FAB): m/z = 273 [M – SCH2CH2COOtBu]+. MS (CI): m/z =
273 [M – SCH2CH2COOtBu]+. HRMS (CI): calcd. for C18H29O9S
[M + H]+ 421.1532; found 421.1521.
2-(tert-Butoxycarbonyl)ethyl 2,3,4-Tri-O-acetyl-1-thio-α-L-fucopyr-
anoside (3α): To a solution of thiofucoside 1α (379 mg, 1.09 mmol)
and tert-butyl 3-bromopropionate (260 µL, 1.56 mmol) in dry
DMF (6 mL) was added Et2NH (0.9 mL). The mixture was stirred
at room temperature for 1 h and then evaporated. Purification of
the residue by flash column chromatography (AcOEt/petroleum
ether, 1:5) afforded 3α (460 mg, 68%) as a white solid. [α]2D0 = –151
2-(Fluoren-9-ylmethoxycarbonylamino)ethyl 2,3,4-Tri-O-acetyl-1-
thio-β-L-fucopyranoside (5β): To a solution of Fmoc-aminoethano-
(c = 0.76, CH Cl ). IR: ν = 3434, 1748, 1643, 1369, 1225,
˜
2
2
1059 cm–1. 1H NMR (300 MHz, CDCl3, 25 °C): δ = 5.70 (d, J1,2
=
thiol (134 mg, 0.45 mmol) and -fucose peracetate (106 mg,
0.34 mmol) in dry dichloromethane (4 mL) was added 4-Å MS, and
the mixture was stirred for 15 min. Then, trimethylsilyl trifluorome-
thanesulfonate (62 µL, 0.45 mmol) was added, and the solution was
stirred at room temperature. After 2 h, the mixture was washed
with saturated aqueous solution of NaHCO3 and water, dried with
Na2SO4, and concentrated. The resulting residue was purified by
column chromatography (AcOEt/petroleum ether, 1:2Ǟ1:1) to give
5β (120 mg, 62%) as a colorless oil. [α]2D0 = –7 (c = 1.2, CH2Cl2).
1H NMR (300 MHz, CDCl3, 25 °C): δ = 7.83–7.31 (m, 5 H, ar-H),
5.55 (br. t, J = 5.4 Hz, 1 H, NHFmoc), 5.20 (m, 2 H, 4-H, 2-H),
5.03 (dd, J2,3 = 9.9 Hz, J3,4 = 3.0 Hz, 1 H 3-H), 4.68 (dd, 2J =
10.5 Hz, 3J = 6.0 Hz, 1 H, -CHH- of Fmoc), 4.47 (dd, 3J = 5.7 Hz,
2J = 10.5 Hz, 1 H, -CHH- of Fmoc), 4.26 (d, J1,2 = 9.6 Hz, 1 H,
1-H), 4.22 (t under 1-H, 1 H, CH of fluorenyl), 3.55 (m, 2 H, 5-H,
1Јa-H), 3.34 (m, 1 H, 1Јb-H, -CH2-), 2.93 (m, 1 H, 2Јa-H) 2.65 (m,
1 H, 2Јb-H), 2.19 (s, 3 H, CH3CO-), 2.08 (s, 3 H, CH3CO-), 2.02
(s, 3 H, CH3CO-), 1.05 (d, J5,CH = 6.3 Hz, 3 H, CH3CO-) ppm.
13C NMR (75 MHz, CDCl3, 25 °C): δ = 170.5, 170.1, 169.7
5.2 Hz, 1 H, 1-H), 5.28 (br. d, J3,4 = 3.2 Hz, 1 H, 4-H), 5.25 (dd,
J2,3 = 10.7 Hz, J1,2 = 5.2 Hz, 1 H, 2-H), 5.18 (dd, J2,3 = 10.7 Hz,
J3,4 = 3.2 Hz, 1 H, 3-H), 4.46 (q, J5,CH = 7.0 Hz, 1 H, 5-H), 2.68–
2.86 (m, 2 H, 1Јa-H, 1Јb-H), 2.54–2.49 (m, 2 H, 2Јa-H, 2Јb-H), 2.16
(s, 3 H, CH3CO-), 2.05 (s, 3 H, CH3CO-), 2.01 (s, 3 H, CH3CO-),
1.45 [s, 9 H, (CH3)3C-], 1.16 (d, J5,CH = 7.0 Hz, 3 H, CH3 of fucose)
ppm. 13C NMR (75 MHz, CDCl3, 25 °C): δ = 170.9, 170.5, 170.2,
169.9 (4 -COO-), 82.6 (1-C), 80.9 [(CH3)C-], 70.9 (4-C), 68.6 (3-C),
68.0 (2-C), 64.8 (5-C), 35.9 (2Ј-C), 28.1 [(CH3)3C-], 25.6 (1Ј-C),
20.8, 20.7, 20.6 (3 CH3CO-), 15.9 (CH3 of fucose) ppm. MS (FAB):
m/z = 273 [M – SCH2CH2COOtBu]+. MS (ESI): m/z = 457
[M + Na]+, 273 [M – SCH2CH2COOtBu]+. MS (CI): m/z = 435
[M + H]+. HRMS (CI): calcd. for C19H31O9S [M + H]+ 435.1689;
found 435.1693.
2-(tert-Butoxycarbonyl)ethyl 2,3,4-Tri-O-acetyl-1-thio-β-L-fucopyr-
anoside (3β): This compound was prepared in the way described
for 3α except that pure 1β was used as starting material. Compound
3β was purified by column chromatography on silica gel (AcOEt/
Eur. J. Org. Chem. 2008, 2973–2982
© 2008 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.eurjoc.org
2977

A. J. Moreno-Vargas, I. Robina et al.
FULL PAPER
(3 -COO-), 156.3 (CO of Fmoc), 143.9, 143.7, 141.3, 127.7, 127.1,
3ЈЈa-H, 3ЈЈb-H), 1.92 (s, 3 H, CH3CO-), 1.05 (br. d, J5,CH = 6.5 Hz,
125.0, 124.7, 120.0 (12 ar-C), 84.2, (1-C), 73.3 (4-C), 72.0 (3-C), 3 H, CH3 of fucose) ppm. 13C NMR (75.4 MHz, [D6]DMSO,
70.2 (2-C), 67.1 (-CH2- of Fmoc), 65.8 (5-C), 47.4 (-CH- of Fmoc),
41.6 (2Ј-C), 31.8 (1Ј-C), 20.8, 20.7, 20.6 (3 CH3CO-), 16.2 (CH3 of
fucose) ppm. MS (FAB): m/z = 594 [M + Na]+. HRMS (FAB):
calcd. for C29H33NO9S [M + Na]+ 594.1803; found 594.1773.
75 °C): δ = 173.9 (-CONH-), 169.7, 168.9, 168.8 (3 CH3COO-),
154.0 (CO of Fmoc), 143.5, 140.3, 128.5, 127.2, 126.8, 126.7, 120.9,
119.5 (6 ar-C), 82.0 (1-C), 71.7 (4-C), 71.3 (5-C), 71.0 (3-C), 70.2
(2-C), 67.3 (4ЈЈ-C, CH2 of Fmoc), 59.1 (2ЈЈ-C), 54.8 (5ЈЈ-C), 46.5
(CH of Fmoc), 39.5 (under DMSO, 3ЈЈ-C), 38.2 (2Ј-C), 29.2 (1Ј-
C), 20.0, 19.8 (3 CH3CO-), 15.6 (CH3 of fucose) ppm. MS (FAB):
m/z = 707 [M + Na]+. HRMS (FAB): calcd. for C34H40N2O11SNa
[M + Na]+ 707.2251; found 707.2258.
2-[(2S,4R)-4-Hydroxy-1-(fluoren-9-ylmethoxycarbonyl)pyrrolidine-
2-carbonylamino]ethyl 2,3,4-Tri-O-acetyl-1-thio-α-L-fucopyranoside
(6α): To a solution of compound 2α (352 mg, 0.782 mmol) in
CH2Cl2 (4 mL) was added TFA (1 mL), and the mixture was stirred
at room temperature for 15 min. After evaporation of the solvent,
the residue was dissolved in DMF (4 mL) and N-Fmoc-4--Hyp-
OH (306 mg, 0.868 mmol), DIEA (0.67 mL, 3.9 mmol), and
PyBOP (442 mg, 0.86 mmol) were sequentially added. After stirring
for 5 h at room temperature, the solvent was removed, and the resi-
due was diluted with CH2Cl2 and washed with 1  HCl, saturated
aqueous solution of NaHCO3, and brine. The organic phase was
then dried (Na2SO4), filtered, and concentrated. Chromatographic
purification on silica gel (ether/ acetone, 7:1) afforded 6α (454 mg,
S-Linked Fucosides 8α and 8β: To a solution of 6α or 6β (345 mg,
0.50 mmol) in DMF (4 mL) was added Et2NH (1 mL), and the
mixture was stirred at room temperature for 20 min. The residue
obtained after evaporation was then dissolved in DMF (4 mL) and
a solution of mono-tert-butyl glutarate (110 mg, 0.6 mmol) in
DMF (4 mL) followed by DIEA (206 µL, 1.2 mmol) and PyBOP
(310 mg, 0.6 mmol) were added. After stirring for 3 h at room tem-
perature, the mixture was evaporated, and the residue was diluted
with CH2Cl2 and washed with 1  HCl, saturated aqueous solution
of NaHCO3, and brine. The organic phase was then dried
1H (Na2SO4), filtered, and concentrated. Chromatographic purifica-
tion on silica gel (ether/acetone, 5:1) afforded 7α (200 mg, 63%) or
7β (56%). Compound 7α or 7β (160 mg, 0.253 mmol) was then
85%) as a colorless oil. [α]2D0 = –95 (c = 1.1, CH Cl ). IR: ν = 3433,
˜
2
2
2972, 2867, 2095, 1747, 1651, 1424, 1360, 1223, 1084 cm–1
.
NMR (500 MHz, [D6]DMSO, 75 °C): δ = 7.85 (d, J = 7.6 Hz, 2 H,
ar-H), 7.64 (d, J = 7.3 Hz, 2 H, ar-H), 7.41 (t, J = 7.3 Hz, 2 H, ar-
H), 7.31 (t, J = 7.3 Hz, 2 H, ar-H), 5.58 (d, J1,2 = 5.0 Hz, 1 H, 1- dissolved in tBuOH/Et3N/H2O (2:1:1, 4 mL), and the mixture was
H), 5.18 (br. s, 1 H, 4-H), 5.13 (dd, J2,3 = 10.8 Hz, J1,2 = 5.0 Hz, 1
H, 2-H), 5.09 (br. d, J2,3 = 10.8 Hz, 1 H, 3-H), 4.4 (br. s, 1 H, 5-
H), 4.35–4.10 (m, 5 H, 2ЈЈ-H, 4ЈЈ-H, -CH2- of Fmoc and -CH- of
stirred at room temperature for 24 h. Evaporation of the solvent
and chromatographic purification (CH2Cl2/MeOH, 8:1) of the resi-
due afforded 8α (90 mg, 70%) as a white foam or 8β (73%). Com-
Fmoc), 3.51 (dd, J5ЈЈa,5ЈЈb = 11.0 Hz, J5ЈЈa,4ЈЈ = 4.6 Hz, 1 H, 5ЈЈa- pound 8β was used in the following step without characterization.
1
H), 3.40 (br. d, J5ЈЈa,5ЈЈb = 11.0 Hz, 1 H, 5ЈЈb-H), 3.25 (m, 2 H, 2Јa-
H, 2Јb-H), 2.61 (m, 2 H, 1Јa-H, 1Јb-H), 2.11 (s, 3 H, CH3CO-),
Data for 8α: [α]2D0 = –177 (c = 1.2, MeOH). H NMR (300 MHz,
CD3OD, mixture of rotamers, 25 °C): Data for major rotamer, δ =
1.98 (s, 3 H, CH3CO-), 1.95 (m, 2 H, 3ЈЈa-H, 3ЈЈb-H), 1.92 (s, 3 H, 5.36 (d, J1,2 = 5.6 Hz, 1 H, 1-H), 4.51–4.43 (m, 2 H, 2ЈЈ-H, 4ЈЈ-H),
CH3CO-), 1.04 (br. s, 3 H, CH3 of fucose) ppm. 13C NMR 4.29 (q, J5,CH = 6.6 Hz, 1 H, 5-H), 4.06 (dd, J2,3 = 10.1 Hz, J1,2
=
(125 MHz, [D6]DMSO, 75 °C): δ = 171.4 (-CONH-), 169.6, 169.1,
168.9 (3 CH3COO-), 143.5, 140.3, 127.2, 126.6, 124.7, 119.6 (6 ar-
5.6 Hz, 1 H, 2-H), 3.75 (dd, J5ЈЈa,5ЈЈb = 10.9 Hz, J5ЈЈa,4ЈЈ = 4.4 Hz, 1
H, 5ЈЈa-H), 3.67 (br. d, J3,4 = 3.2 Hz, 1 H, 4-H), 3.60 (dd, J2,3
=
C), 81.6 (1-C), 70.2 (4-C), 67.7, 67.6, 67.1, 66.4 (2-C, 3-C, 4ЈЈ-C, 10.1 Hz, J3,4 = 3.2 Hz, 1 H, 3-H), 3.52 (br. d, J5ЈЈa,5ЈЈb = 10.9 Hz, 1
CH2 of Fmoc), 64.5 (5-C), 58.7 (2ЈЈ-C), 54.3 (5ЈЈ-C), 46.5 (CH of
Fmoc), 39.5 (3ЈЈ-C), 38.4 (2Ј-C), 28.8 (1Ј-C), 19.9, 19.8, 19.7 (3
CH3CO-), 15.1 (CH3 of fucose) ppm. MS (FAB): m/z = 707 [M +
Na]+. HRMS (FAB): calcd. for C34H40N2O11SNa [M + Na]+
707.2251; found 707.2269.
H, 5ЈЈb-H), 3.43 (m, 2 H, 2Јa-H, 2Јb-H), 2.77–2.65 (m, 2 H, 1Јa-
H, 1Јb-H), 2.42 (t, J2ЈЈЈ,3ЈЈЈ = 7.6 Hz, 2 H, 2ЈЈЈ-H), 2.32 (t, J3ЈЈЈ,4ЈЈЈ
= 7.4 Hz, 2 H, 4ЈЈЈ-H), 2.27–1.83 (m, 4 H, 3ЈЈa-H, 3ЈЈb-H, 3ЈЈЈ-H),
1.46 [s, 9 H, (CH3)3C-], 1.24 (d, J5,CH = 6.6 Hz, 3 H, -CH3 of
fucose) ppm. 13C NMR (75 MHz, CD3OD, 25 °C mixture of rota-
mers): Data for major rotamer, δ = 175.1, 175.0, 174.7 (3 -CO),
88.4 (1-C), 82.0 [-C(CH3)3], 73.9 (4-C), 72.9 (3-C), 71.3 (4ЈЈ-C), 70.0
(2-C), 68.7 (5-C), 60.9 (2ЈЈ-C), 57.0 (5ЈЈ-C), 41.0 (2Ј-C), 39.8 (3ЈЈ-
C), 36.0 (4ЈЈЈ-C), 35.0 (2ЈЈЈ-C), 31.1 (1Ј-C), 28.9 [(CH3)3C-], 21.8
(3ЈЈЈ-C), 17.1 (-CH3 of fucose) ppm. MS (FAB): m/z = 529 [M +
Na]+. HRMS (FAB): calcd. for C22H38N2O9SNa [M + Na]+
529.2195; found 529.2196.
2-[(2S,4R)-4-Hydroxy-1-(fluoren-9-ylmethoxycarbonyl)pyrrolidine-
2-carbonylamino]ethyl 2,3,4-Tri-O-acetyl-1-thio-α-L-fucopyranoside
(6β): To a solution of compound 5β (68 mg, 0.109 mmol) in DMF
(2 mL) was added Et2NH (0.4 mL), and the mixture was stirred at
room temperature for 15 min. After evaporation of the solvent, the
residue was dissolved in DMF (2 mL) and N-Fmoc-4--Hyp-OH
(42 mg, 0.12 mmol), DIEA (41 µL, 0.24 mmol), and PyBOP
(62 mg, 0.12 mmol) were sequentially added. After stirring for 5 h
[(2S,4R)-1-(4-Carboxybutanoyl)-4-hydroxypyrrolidine-2-carbonyl-
at room temperature, the solvent was removed, and the residue was amino]ethyl 1-Thio-α and β-L-Fucopyranoside (9α and 9β): A 20%
diluted with CH2Cl2 and washed with 1  HCl, saturated aqueous
solution of NaHCO3, and brine. The organic phase was then dried
(Na2SO4), filtered, and concentrated. Chromatographic purifica-
tion on silica gel (ether/acetone, 7:1) afforded 6β (50 mg, 67%) as
solution of TFA in CH2Cl2 (5 mL) was added to compound 8α
(65 mg, 0.128 mmol) or 8β, and the mixture was stirred at room
temperature for 20 min. Evaporation of the solvent afforded 9α
(55 mg, quant.) or 9β (quant.) as a white solid. Data for 9α: [α]2D0
1
1
a colorless oil. [α]2D0 = –13 (c = 1.2, CH2Cl2). H NMR (300 MHz,
= –59 (c = 0.75, H2O). H NMR (300 MHz, D2O, 25 °C mixture
[D6]DMSO, 75 °C): δ = 7.93 (br. s, 1 H, -NH-), 7.90 (d, J = 7.6 Hz,
2 H, ar-H), 7.84 (d, J = 7.3 Hz, 2 H, ar-H), 7.44 (t, J = 7.3 Hz, 2
H, ar-H), 7.34 (t, J = 7.3 Hz, 2 H, ar-H), 5.15 (m, 2 H, 4-H, 3-H),
4.97 (t, J2,3 = J1,2 = 9.6 Hz, 1 H, 2-H), 4.90 (br. s, 1 H, -OH), 4.76
of rotamers): Data for major rotamer, δ = 5.36 (d, J1,2 = 5.6 Hz, 1
H, 1-H), 4.51 (m, 1 H, 4ЈЈ-H), 4.39 (t, J2ЈЈ,3ЈЈa = J2ЈЈ,3ЈЈb = 8.6 Hz,
1 H, 2ЈЈ-H), 4.28 (q, J5,CH = 6.6 Hz, 1 H, 5-H), 4.00 (dd, J2,3
=
10.4 Hz, J1,2 = 5.6 Hz, 1 H, 2-H), 3.74 (m, 2 H, 5ЈЈa-H, 4-H), 3.64
(d, J1,2 = 9.6 Hz, 1 H, 1-H), 4.41–4.16 (m, 5 H, 2ЈЈ-H, 4ЈЈ-H, -CH2- (dd, J2,3 = 10.4 Hz, J3,4 = 3.3 Hz, 1 H, 3-H), 3.58 (br. d, J5ЈЈa,5ЈЈb
=
of Fmoc and -CH- of Fmoc), 4.02 (br. q, J5,CH = 6.5 Hz, 1 H, 5-
H), 3.52 (dd, J5ЈЈa,5ЈЈb = 11.0, J5ЈЈa,4 = 4.5, 5ЈЈa-H), 3.42–3.24 (m, 3
H, 5ЈЈb-H, 2Јa-H, 2Јb-H), 2.75 (m, 1 H, 1Јa-H), 2.63 (m, 1 H, 1Јb-
11.6 Hz, 1 H, 5ЈЈb-H), 3.44–3.32 (m, 2 H, 2Јa-H, 2Јb-H), 2.78–2.65
(m, 2 H, 1Јa-H, 1Јb-H), 2.44–1.79 (m, 8 H, 2ЈЈЈ-H, 3ЈЈЈ-H, 4ЈЈЈ-H,
3ЈЈa-H, 3ЈЈb-H), 2.32 (t, J3ЈЈЈ,4ЈЈЈ = 7.4 Hz, 2 H, 4ЈЈЈ-H), 2.27–1.83
H), 2.12 (s, 3 H, CH3CO-), 1.99 (s, 3 H, CH3CO-), 1.95 (m, 2 H, (m, 4 H, 3ЈЈa-H, 3ЈЈb-H, 3ЈЈЈ-H), 1.16 (d, J5,CH = 6.6 Hz, 3 H,
2978
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© 2008 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Eur. J. Org. Chem. 2008, 2973–2982

S-Neofucopeptides as E- and P-Selectin Inhibitors
-CH3 of fucose) ppm. 13C NMR (75 MHz, D2O, 25 °C, mixture of
rotamers): Data for major rotamer, δ = 180.0 (-COOH), 176.6,
NMR (75 MHz, CD3OD, 25 °C): δ = 165.4 (COOEt), 160.7 (5-C),
149.2 (2-C), 160.2, 146.1, 136.9, 131.7, 129.7, 128.7, 127.9, 114.0
176.0 (2 -CONH-), 88.2 (1-C), 73.6 (4-C), 72.2 (3-C), 71.6 (4ЈЈ-C), (18 ar-C), 115.3 (3-C), 111.8 (4-C), 87.8 [(Ar)3C-], 74.7 (2Ј-C), 72.6
69.6, (2-C), 69.3 (5-C), 61.1 (2ЈЈ-C), 57.4 (5ЈЈ-C), 41.1 (2Ј-C), 39.4 (3Ј-C), 66.5 (4Ј-C), 61.4 (-CH2CH3), 60.9 (1Ј-C), 55.7 (-OCH3), 14.7
(3ЈЈ-C), 35.0 (4ЈЈЈ-C), 34.8 (2ЈЈЈ-C), 31.5 (1Ј-C), 21.5 (3ЈЈЈ-C), 17.3 (-CH2CH3), 13.8 (CH3 of furan) ppm. MS (FAB): m/z = 594 [M +
(-CH3 of fucose) ppm. MS (FAB): m/z = 473 [M + Na]+. HRMS Na]+. HRMS (FAB) calcd. for C32H33N3O7Na [M + Na]+
(FAB) calcd. for C18H30N2O9SNa [M + Na]+ 473.1570; found
473.1581. Data for 9β: [α]2D0 = –10 (c = 1.4, MeOH). 13C NMR
(75 MHz, CD3OD, 25 °C, mixture of rotamers): Data for major
rotamer, δ = 177.0 (-COOH), 174.7, 174.2 (2 -CONH-), 87.5 (1-C),
76.4 (4-C), 76.2 (3-C), 73.3 (4ЈЈ-C), 71.2 (2-C), 70.8 (5-C), 60.5 (2ЈЈ-
C), 56.6 (5ЈЈ-C), 40.0 (2Ј-C), 39.3 (3ЈЈ-C), 34.5 (4ЈЈЈ-C), 33.9 (2ЈЈЈ-
C), 30.5 (1Ј-C), 21.2 (3ЈЈЈ-C), 17.1 (-CH3 of fucose). MS (FAB):
m/z (%) = 473 (100) [M + Na]+. HRMS (FAB) calcd. for
C18H30N2O9SNa [M + Na]+ 473.1570; found 473.1588.
594.2216; found 594.2211.
Ethyl 5-(1-Azido-1-deoxy-D-arabino-tetritol-1-yl)-2-methylfuran-3-
carboxylate (17): Compound 16 (826 mg, 1.45 mmol) was treated
with a solution of TFA/CH2Cl2/triisopropylsilane (TIPS) (93:2:5)
(15 mL) at room temperature for 30 min. After evaporation of the
solvent and chromatographic purification (CH2Cl2/MeOH, 15:1),
compound 17 (345 mg, 80%) was obtained as a colorless oil. [α]2D0
= +86 (c = 1.5, CH Cl ). IR: ν = 3412, 2106, 1694, 1230, 1082 cm–1.
˜
2
2
1H NMR (300 MHz, CD3OD, 25 °C): δ = 6.78 (s, 1 H, 4-H), 4.67
3
Ethyl 5-[4-O-(p-Methoxytrityl)-D-arabino-tetritol-1-yl]-2-methyl-
(d, J1Ј,2Ј = 4.1 Hz, 1 H, 1Ј-H), 4.28 (q, JH,H = 7.2 Hz, 2 H,
furan-3-carboxylate (13): To a solution of 12[26] (1 g, 3.65 mmol) in
pyridine (8 mL) cooled to 0 °C was added MeOTrCl (1.4 g,
4.38 mmol). After 3 h at room temperature MeOH (1.5 mL) was
added, and the mixture was stirred for 15 min. The residue ob-
tained after evaporation of the solvent was diluted with CH2Cl2
and washed with water. The organic phase was then dried
(Na2SO4), filtered, and concentrated. Chromatographic purifica-
tion on silica gel (CH2Cl2/acetone, 30:1, 1 % Et3N) afforded 13
-CH2CH3), 3.93 (dd, J2Ј,3Ј = 8.1 Hz, J1Ј,2Ј = 4.1 Hz, 1 H, 2Ј-H),
3.76 (dd, J4Јa,4Јb = 11.4 Hz, J3Ј,4Јa = 3.5 Hz, 1 H, 4Јa-H), 3.61 (dd,
J4Јa,4Јb = 11.4 Hz, J3Ј,4Јb = 5.8 Hz, 1 H, 4Јb-H), 3.46 (ddd, J2Ј,3Ј
=
8.1 Hz, J3,4Јb = 5.8 Hz, J3Ј,4Јa = 3.5 Hz, 1 H, 3Ј-H), 2.57 (s, 3 H,
3
CH3 of furan), 1.35 (t, JH,H = 7.2 Hz, 3 H, -CH2CH3) ppm. 13C
NMR (75 MHz, CD3OD, 25 °C): δ = 165.4 (COOEt), 160.6 (2-C),
149.2 (5-C), 115.3 (3-C), 111.8 (4-C), 74.6 (2Ј-C), 73.4 (3Ј-C), 64.5
(4Ј-C), 61.4 (-CH2CH3), 60.9 (1Ј-C), 14.6 (-CH2CH3), 13.8 (CH3 of
furan) ppm. MS (CI) m/z = 257 [M – N3]+. HRMS (CI): calcd. for
C12H18O6N3 [M + H]+ 300.1196; found 300.1199.
(1.59 g, 81%) as a yellow oil. [α]2D0 = +15 (c = 1, CH Cl ). IR: ν =
˜
2
2
3846, 3752, 3472, 2928, 1703, 1508, 1449, 1236, 1080 cm–1
.
1H
NMR (300 MHz, CDCl3, 25 °C): δ = 7.43–6.83 (m, 14 H, ar-H),
6.52 (s, 1 H, 4-H), 4.74 (d, J1Ј,2Ј = 3.4 Hz, 1 H, 1Ј-H), 4.27 (q, 3JH,H
= 7.1 Hz, 2 H, -CH2CH3), 3.93 (dd, J2Ј,3Ј = 5.9 Hz, J1Ј,2Ј = 3,4 Hz,
1 H, 2Ј-H), 3.83 (m, 1 H, 3Ј-H), 3.80 (s, 3 H, -OCH3), 3.41 (dd,
Ethyl 5-(1-Amino-1-deoxy-D-arabino-tetritol-1-yl)-2-methylfuran-3-
carboxylate (18): A solution of 17 (340 mg, 1.14 mmol) in absolute
EtOH (20 mL) was hydrogenated under atmospheric pressure for
1 h by using Pd/C (10%) as a catalyst. Then, the solution was fil-
tered through Celite, and the catalyst was washed with EtOH. The
filtered solution was concentrated in vacuo to give pure 18 (307 mg,
1.12 mmol, 98%) as a colorless oil. [α]2D0 = –11 (c = 0.57, MeOH).
1H NMR (300 MHz, CD3OD, 25 °C): δ = 6.58 (s, 1 H, 4-H), 4.26
J4Јa,4Јb = 9.9 Hz, J3Ј,4Јa = 4.3 Hz, 1 H, 4Јa-H), 3.30 (dd, J4Јa,4Јb
=
9.9 Hz, J3Ј,4Јb = 5.8 Hz, 1 H, 4Јb-H), 2.72 (br. s, 2 H, -OH), 2.54
3
(s, 3 H, CH3- of furan), 1.66 (br. s, 1 H, -OH), 1.33 (t, JH,H
=
7.1 Hz, 3 H, -CH2CH3) ppm. 13C NMR (75 MHz, CDCl3, 25 °C):
δ = 164.1 (COOEt), 159.1 (ar-C), 158.9 (2-C), 151.8 (5-C), 144.1,
144.0, 135.2, 130.4, 128.4, 128.2, 127.3, 113.5 (17 ar-C), 114.4 (3-
C), 108.7 (4-C), 87.2 [(Ar)3C-], 73.7 (2Ј-C), 71.1 (3Ј-C), 67.3 (1Ј-C),
6 4 . 8 ( 4 Ј - C ) , 6 0 . 3 ( - C H 2 C H 3 ) , 5 5 . 4 ( - O C H 3 ) , 1 4 . 5
(-CH2CH3), 13.9 (CH3- of furan) ppm. MS (FAB): m/z = 569 [M
+ Na]+. HRMS (FAB): calcd. for C32H34O8Na [M + Na]+
569.2151; found 569.2171.
3
(q, JH,H = 7.1 Hz, 2 H, -CH2CH3), 4.23 (d, J1Ј,2Ј = 4.5 Hz, 1 H,
1Ј-H), 3.75 (dd, J4Јa,4Јb = 11.4 Hz, J3Ј,4Јa = 3.5 Hz, 1 H, 4Јa-H),
3.74 (dd, J2Ј,3Ј = 8.7 Hz, J1Ј,2Ј = 4.5 Hz, 1 H, 2Ј-H), 3.60 (dd, J4Јa,4Јb
= 11.4 Hz, J3Ј,4Јb = 5.5 Hz, 1 H, 4Јb-H), 3.42 (ddd, J2Ј,3Ј = 8.7 Hz,
J3Ј,4Јb = 5.5 Hz, J3Ј,4Јa = 3.5 Hz, 1 H, 3Ј-H), 2.55 (s, 3 H, CH3 of
3
furan), 1.33 (t, JH,H = 7.1 Hz, 3 H, -CH2CH3) ppm. 13C NMR
(75 MHz, CD3OD, 25 °C): δ = 165.7 (COOEt), 159.9 (2-C), 153.6
(5-C), 115.2 (3-C), 109.3 (4-C), 74.5 (2Ј-C), 74.2 (3Ј-C), 64.8 (4Ј-
C), 61.3 (-CH2CH3), 52.6 (1Ј-C), 14.7 (-CH2CH3), 13.8 (CH3 of
f u r a n ) p p m . M S ( C I ) : m/ z = 2 7 4 [ M – H] + , 2 5 7 [ M –
NH3]+. HRMS (CI): calcd. for C12H20O6N [M – H]+ 274.1291;
found 274.1296.
Ethyl 5-[1-Azido-1-deoxy-4-O-(p-methoxytrityl)-D-arabino-tetritol-
1-yl]-2-methylfuran-3-carboxylate (16): To a cooled solution of 13
(1.163 g, 1.96 mmol) in dry CH2Cl2 (6 mL) was added Et3N
(539 µL, 8.62 mmol) and a solution of SOCl2 (350 µL, 4.9 mmol)
in CH2Cl2 (1 mL). After 30 min at 0 °C, cold diethyl ether was
added, and the mixture was washed with water and brine. The or-
ganic phase was then dried (Na2SO4), filtered, and concentrated.
The mixture of sulfites 14 and 15 thus obtained was dissolved in
THF (7 mL) and TMSN3 (885 µL, 6.39 mmol) and TBAF (1 /
THF, 6.39 mL) were sequentially added. After 3 d at room tem-
perature, the solvent was evaporated, and the residue was purified
by column chromatography (ether/petroleum ether, 1:1, 1% Et3N)
to afford 16 (850 mg, 76%) as a colorless oil. [α]2D0 = +39 (c = 0.9,
Ethyl 5-(1-Azido-1-deoxy-4-O-tosyl-D-arabino-tetritol-1-yl)-2-meth-
ylfuran-3-carboxylate (19): To a solution of compound 17 (107 mg,
0.357 mmol) in pyridine (1.5 mL) at –15 °C was added TsCl
(204 mg, 1.071 mmol) in portions. The mixture was stirred from
–15 to 5 °C for 7 h. Then, water was added and the solvent was
evaporated. The residue was diluted with CH2Cl2 and sequentially
washed with 1  HCl, saturated aqueous solution of NaHCO3, and
brine. The organic phase was dried (Na2SO4), filtered, and concen-
CH Cl ). IR: ν = 3482, 2933, 2102, 1714, 1609, 1577, 1509, 1446,
˜
2
2
1299, 1251, 1229, 1179, 1079, 1033, 902, 831, 796, 777, 707, trated. Chromatographic purification on silica gel (ether/petroleum
632 cm–1. H NMR (300 MHz, CD3OD, 25 °C): δ = 7.32–6.84 (m, ether, 1:2Ǟ1:1) afforded 19 (118 mg, 73%). [α]2D0 = +74 (c = 0.81,
1
14 H, ar-H), 6.71 (s, 1 H, 4-H), 4.62 (d, J1Ј,2Ј = 4.4 Hz, 1 H, 1Ј-H), CH Cl ). IR: ν = 3468, 2983, 2925, 2106, 1713, 1360, 1230, 1175,
˜
2
2
3
4.28 (q, JH,H = 7.2 Hz, 2 H, -CH2CH3), 3.97 (dd, J2Ј,3Ј = 7.4 Hz,
1094, 980, 810, 775 cm–1. 1H NMR (300 MHz, CDCl3, 25 °C): δ =
J1Ј,2Ј = 4.4 Hz, 1 H, 2Ј-H), 3.80 (s, 3 H, -OCH3), 3.60 (m, 1 H, 3Ј- 7.79 (d, J = 8.4 Hz, 2 H, ar-H), 7.35 (d, J = 8.4 Hz, 2 H, ar-H),
H), 3.35 (dd, J4Јa,4Јb = 9.8 Hz, J3Ј,4Јa = 3.4 Hz, 1 H, 4Јa-H), 3.22 6.74 (s, 1 H, 4-H), 4.75 (d, J1Ј,2Ј = 4.5 Hz, 1 H, 1Ј-H), 4.27 (q, 3JH,H
(dd, J3Ј,4Јb = 6.2 Hz, J4Јa,4Јb = 9.8 Hz, 1 H, 4Јb-H), 2.56 (s, 3 H, = 7.2 Hz, 2 H, -CH2CH3), 4.23 (m, 2 H, 4Јa-H, 4Јb-H), 3.96 (dd,
3
CH3 of furan), 1.35 (t, JH,H = 7.2 Hz, 3 H, -CH2CH3) ppm. 13C J2Ј,3Ј = 7.8 Hz, J1Ј,2Ј = 4.5 Hz, 1 H, 2Ј-H), 3.72 (m, 1 H, 3Ј-H), 2.64
Eur. J. Org. Chem. 2008, 2973–2982
© 2008 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.eurjoc.org
2979

A. J. Moreno-Vargas, I. Robina et al.
FULL PAPER
(br. s, 2 H, OH), 2.57 (s, 3 H, CH3 of furan), 2.45 (s, 3 H, CH3 of
= 2.4 Hz, 1 H, 2ЈЈ-H), 3.54 (m, 2 H, 3-H, 5-H), 3.47 (dd, J4ЈЈa,4ЈЈb
= 14.1 Hz, J4ЈЈa,3ЈЈ = 3.3 Hz, 1 H, 4ЈЈa-H), 3.39 (d, J1,2 = 9.0 Hz, 1
H, 2-H), 3.35 (dd, J = 8.6 Hz, J = 3.4 Hz, 1 H, 4-H), 3.23 (dd
3
Ts), 1.34 (t, JH,H = 7.2 Hz, 3 H, -CH2CH3) ppm. 13C NMR
(75 MHz, CDCl3, 25 °C): δ = 163.6 (COOEt), 160.0 (2-C), 146.3
(5-C), 145.3 (1-C of Ph), 132.3 (4-C of Ph), 130.0 (2 ar-C), 129.0 under CD3OD, 1 H, 4ЈЈb-H), 2.84 (m, 2 H, 2Јa-H, 2Јb-H), 2.49 (t,
(2 ar-C), 114.4 (3-C), 111.5 (4-C), 72.3 (2Ј-C), 71.1 (4Ј-C), 70.0 (3Ј-
C), 60.4 (-CH2CH3), 59.6 (1Ј-C), 21.6 (Me of Ts), 14.3 (-CH2CH3),
13.9 (CH3 of furan) ppm. MS (FAB): m/z = 476 [M + Na]+. HRMS
(FAB): calcd. for C19H23N3O8SNa [M + Na]+ 476.1104; found
476.1105.
J3Јa,2Јa = J3Јa,2Јb = J3Јb,2Јa = J3Јb,2Јb = 7.2 Hz, 2 H, 3Јa-H, 3Јb-H),
2.44 (s, 3 H, CH3 of furan), 1.23 (t, 3JH,H = 6.9 Hz, 3 H, -CH2CH3),
1.16 (d, J5,CH = 6.6 Hz, 3 H, CH3 of fucose) ppm. 13C NMR
(75 MHz, CD3OD, 25 °C): δ = 175.7 (COOEt), 166.2 (1Ј-C), 160.0,
155.9 (2ЈЈЈ-C, 5ЈЈЈ-C), 115.6 (3ЈЈЈ-C), 109.0 (4ЈЈЈ-C), 87.9 (1-C),
76.9, 76.6 (3-C, 4-C), 75.3 (2ЈЈ-C), 73.7 (5-C), 71.6, 71.5 (3ЈЈ-C, 2-
C), 68.2 (1ЈЈ-C), 61.8 (-CH2CH3), 44.5 (4ЈЈ-C), 38.5 (3Ј-C), 27.5 (2Ј-
C), 17.6 (CH3 of fucose), 15.1 (-CH2CH3), 14.3 (CH3 of furan)
ppm. MS (FAB): m/z = 530 [M + Na]+. HRMS (FAB): calcd. for
C21H34NO11SNa [M + Na]+ 530.1672; found 530.1679.
Ethyl 5-[(2R,3S,4R)-3,4-Dihydroxypyrrolidin-2-yl]-2-methylfuran-3-
carboxylate (20):[27] A solution of 19 (269 mg, 0.593 mmol) in abso-
lute EtOH (7 mL) was hydrogenated with Pd-C (10%) as catalyst.
After 1.5 h, the catalyst was filtered off and the solution was con-
centrated to afford 20 (250 mg, quant.) as a white solid.
Compound 23α: This compound was prepared as described for 21α
except that pure thiofucoside 4α (291 mg, 0.69 mmol) and amino-
ester 18 (189 mg, 0.69 mmol) were used as starting materials. After
purification by column chromatography on silica gel (CH2Cl2/
MeOH, 15:1), compound 23α (284 mg, 66%) was obtained as a
colorless oil. This compound was used in the following step without
characterization.
S-Linked Fucoside 21α: Compound 3α (57 mg, 0.131 mmol) was
treated with TFA (25% in CH2Cl2, 2.5 mL) at room temperature
over 30 min, and the solvent was then evaporated. The residue was
dissolved in DMF (3 mL) and DIEA (98 µL, 0.576 mmol), com-
pound 11 (40 mg, 0.144 mmol), and PyBOP (74 mg, 0.144 mmol)
were sequentially added. After stirring for 2 h at room temperature,
the solvent was removed, and the residue was diluted with AcOEt
and washed with 1  HCl, saturated aqueous solution of NaHCO3,
and brine. The organic phase was then dried (Na2SO4), filtered,
and concentrated. Chromatographic purification on silica gel
(CH2Cl2/MeOH, 20:1) afforded 21α (80 mg, 96%), which was used
in the following step without characterization.
Compound 24α: This compound was prepared as described for 22α
except that pure thiofucoside 23α (256 mg, 0.41 mmol) was used as
the starting material. After purification by column chromatography
on silica gel (CH2Cl2/MeOH, 6:1), compound 24α (112 mg, 55%)
was obtained as a white foam. [α]2D0 = –71 (c = 0.41, H2O). 1H
NMR (300 MHz, D2O, 25 °C): δ = 6.68 (s, 1 H, 4ЈЈЈ-H), 5.44 (d,
J1,2 = 5.4 Hz, 1 H, 1-H), 5.26 (d, J1ЈЈ,2ЈЈ = 4.2 Hz, 1 H, 1ЈЈ-H), 4.30
Compound 21β: This compound was prepared as described for 21α
by using 3β (70 mg, 0.161 mmol) as the starting material. After
purification by column chromatography on silica gel (CH2Cl2/
MeOH, 30:1), compound 21β (80 mg, 82%) was obtained as a col-
orless oil. This compound was used in the following step without
characterization.
3
(q, JH,H = 7.2 Hz, 2 H, -CH2CH3), 4.13 (q, J5,CH = 6.6 Hz, 1 H,
5-H), 4.07 (dd, J2,3 = 9.3 Hz, J1,2 = 5.4 Hz, 1 H, 2-H), 3.88 (dd,
J2ЈЈ,3ЈЈ = 8.4 Hz, J1ЈЈ,2ЈЈ = 4.2 Hz, 1 H, 2ЈЈ-H), 3.79–3.69 (m, 3 H,
4ЈЈa-H, 3-H, 4-H), 3.62 (dd, J4ЈЈa,4ЈЈb = 11.9 Hz, J4ЈЈb,3ЈЈ = 6.6 Hz, 1
H, 4ЈЈb-H), 3.50 (m, 1 H, 3ЈЈ-H), 3.48 (d, J1Јa,1Јb = 15.3 Hz, 1 H,
1Јa-H), 3.28 (d, J1Јa,1Јb = 15.3 Hz, 1 H, 1Јb-H), 2.55 (s, 3 H, CH3
Compound 22α: Compound 21α (60 mg, 0.095 mmol) was dissolved
in EtOH/Et3N/H2O (2:1:1, 4 mL), and the mixture was stirred at
room temperature for 24 h. Evaporation of the solvent and chro-
matographic purification (CH2Cl2/MeOH, 6:1) of the residue af-
forded 22α (35 mg, 74%) as a white foam. [α]2D0 = –89 (c = 0.8,
MeOH). 1H NMR (300 MHz, CD3OD, 25 °C): δ = 6.60 (s, 1 H,
4ЈЈЈ-H), 5.37 (d, J1,2 = 5.6 Hz, 1 H, 1-H), 4.87 (d under H2O, 1 H,
3
of furan), 1.34 (t, JH,H = 7.2 Hz, 3 H, -CH2CH3), 1.07 (d, J5,CH
= 6.6 Hz, 3 H, -CH3 of fucose) ppm. 13C NMR (75 MHz, D2O): δ
= 171.9 (-COOEt), 166.3 (-CONH-), 160.3, 148.6 (2ЈЈЈ-C, 5ЈЈЈ-C),
113.5 (3ЈЈЈ-C), 109.0 (4ЈЈЈ-C), 86.8 (1-C), 71.9 (2ЈЈ-C), 71.6 (4-C),
71.5 (3ЈЈ-C), 70.1 (3-C), 67.6 (2-C, 5-C), 61.5 (-CH2CH3), 49.2 (1ЈЈ-
C), 33.7 (1Ј-C), 15.1 (CH3 of fucose), 13.4, 13.2 (-CH2CH3, CH3
of furan) ppm. MS (FAB): m/z = 516 [M + Na]+. HRMS (FAB):
calcd. for C20H31NO11SNa [M + Na]+ 516.1516; found 516.1541.
3
1ЈЈ-H), 4.28 (q, JH,H = 7.1 Hz, 2 H, -CH2CH3), 4.25 (m, 1 H, 5-
H), 4.05 (dd, J2,3 = 10.1 Hz, J1,2 = 5.6 Hz, 1 H, 2-H), 3.78 (m, 1
H, 3ЈЈ-H), 3.68 (m, 2 H, 2ЈЈ-H, 4-H), 3.58 (m, 2 H, 4ЈЈa-H, 3-H),
3.35 (dd under CD3OD, J4ЈЈb,3ЈЈ = 6.6 Hz, 1 H, 4ЈЈb-H), 2.82 (m, 2
Compound 25α: This compound was prepared as described for 21α
except that pure thiofucoside 3α (94 mg, 0.216 mmol) and amino-
ester 18 (59 mg, 0.216 mmol) were used as starting materials. After
purification by column chromatography on silica gel (CH2Cl2/
MeOH, 20:1), compound 25α (103 mg, 75%) was obtained as a
colorless oil. This compound was used in the following step without
characterization.
H, 2Јa-H, 2Јb-H), 2.58 (t, J3Јa,2Јa = J3Јa,2Јb = J3Јb,2Јa = J3Јb,2Јb
=
7.1 Hz, 2 H, 3Јa-H, 3Јb-H), 2.55 (s, 3 H, CH3 of furan), 1.35 (t,
3JH,H = 7.1 Hz, 3 H, -CH2CH3), 1.24 (d, J5,CH = 6.7 Hz, 3 H, CH3
of fucose) ppm. 13C NMR (75 MHz, CD3OD, 25 °C): δ = 173.7
(COOEt), 164.3 (1Ј-C), 158.2, 154.0 (2ЈЈЈ-C, 5ЈЈЈ-C), 113.7 (3ЈЈЈ-C),
107.1 (4ЈЈЈ-C), 86.2 (1-C), 73.4, 72.0 (2ЈЈ-C, 4-C), 71.0 (2-C), 69.8
(3ЈЈ-C), 68.1 (2-C), 66.8 (5-C), 66.3 (1ЈЈ-C), 59.9 (1ЈЈ-C), 42.6 (4ЈЈ-
C), 35.9 (3Ј-C), 25.6 (2Ј-C), 15.2 (CH3 of fucose), 13.2 (-CH2CH3),
12.4 (CH3 of furan) ppm. MS (FAB): m/z = 530 [M + Na]+. HRMS
(FAB): calcd. for C21H34NO11SNa [M + H]+ 508.1853; found
508.1840.
Compound 26α: This compound was prepared as described for 22α
except that pure thiofucoside 25α (60 mg, 0.094 mmol) was used as
the starting material. Purification by column chromatography on
silica gel (CH2Cl2/MeOH, 8:1) afforded 26α (29 mg, 60%) as a
white foam. [α]2D0 = –64 (c = 1.3, MeOH). IR: ν = 3409, 2936, 1654,
˜
Compound 22β: This compound was prepared as described for 22α
by using 21β (43 mg, 0.068 mmol) as the starting material. After
purification by column chromatography on silica gel (CH2Cl2/
MeOH, 6:1), compound 22β (26 mg, 74%) was obtained as a white
foam. [α]2D0 = +15 (c = 0.4, MeOH). 1H NMR (300 MHz, CD3OD,
25 °C): δ = 6.49 (s, 1 H, 4ЈЈЈ-H), 4.77 (d, J1ЈЈ,2ЈЈ = 2.4 Hz, 1 H, 1ЈЈ-
1534, 1428, 1299, 1089, 1028, 780 cm–1. 1H NMR (300 MHz,
CD3OD, 25 °C): δ = 6.61 (s, 1 H, 4ЈЈЈ-H), 5.36 (d, J1ЈЈ,2ЈЈ = 4.2 Hz,
3
1 H, 1ЈЈ-H), 5.34 (d, J1,2 = 5.7 Hz, 1 H, 1-H), 4.27 (q, JH,H
7.2 Hz, 2 H, -CH2CH3), 4.21 (m, 1 H, 5-H), 4.03 (dd, J2,3
=
=
10.0 Hz, J1,2 = 5.7 Hz, 1 H, 2-H), 3.80 (dd, J2ЈЈ,3ЈЈ = 8.1 Hz, J1ЈЈ,2ЈЈ
= 4.2 Hz, 1 H, 2ЈЈ-H), 3.74 (dd, J4ЈЈa,4ЈЈb = 11.4 Hz, J4ЈЈa,3ЈЈ = 3.3 Hz,
1 H, 4ЈЈa-H), 3.63–3.54 (m, 3 H, 4ЈЈb-H, 3-H, 4-H), 3.45 (m, 1 H,
3
H), 4.22 (d, J1,2 = 9.0 Hz, 1 H, 1-H), 4.16 (q, JH,H = 6.9 Hz, 2 H,
-CH2CH3), 3.66 (m, 1 H, 3ЈЈ-H), 3.56 (dd, J2ЈЈ,3ЈЈ = 6.9 Hz, J1ЈЈ,2ЈЈ 3ЈЈ-H), 2.82 (m, 2 H, 1Јa-H, 1Јb-H), 2.57 (m, 2 H, 2Јa-H, 2Јb-H),
2980
www.eurjoc.org
© 2008 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Eur. J. Org. Chem. 2008, 2973–2982

S-Neofucopeptides as E- and P-Selectin Inhibitors
2.54 (s, 3 H, CH3 of furan), 1.33 (t, 3JH,H = 7.2 Hz, 3 H, -CH2CH3),
1.21 (d, J5,CH = 6.6 Hz, 3 H, -CH3 of fucose) ppm. 13C NMR
(75 MHz, CD3OD, 25 °C): δ = 173.4 (-COOEt), 165.7 (-CONH-),
159.8, 151.6 (2ЈЈЈ-C, 5ЈЈЈ-C), 115.1 (3ЈЈЈ-C), 109.9 (4ЈЈЈ-C), 87.7 (1-
C), 74.3 (2ЈЈ-C), 73.4 (4-C), 73.3 (3ЈЈ-C), 72.4 (3-C), 69.5 (2-C), 68.2
(5-C), 64.6 (4ЈЈ-C), 61.3 (-CH2CH3), 50.4 (1ЈЈ-C), 37.3 (2Ј-C), 26.9
(1Ј-C), 16.6 (CH3 of fucose), 14.7 (-CH2CH3), 13.9 (CH3 of furan)
ppm. MS (FAB): m/z = 530 [M + Na]+. HRMS (CI): calcd. for
C21H34NO11S [M + H]+ 508.1853; found 508.1871.
[7]
a) C. Filser, D. Kowalczyk, C. Jones, M. K. Wild, U. Ipe, D.
Vestweber, H. Kunz, Angew. Chem. Int. Ed. 2007, 46, 2108–
2111; b) R. W. Denton, X. Cheng, K. A. Tony, A. Dilhas, J. J.
Hernández, A. Canales, J. Jiménez-Barbero, D. R. Mootoo,
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e) S. M. Chervin, J. B. Lowe, M. Koreeda, J. Org. Chem. 2002,
67, 5654–5662.
For selected references, see: a) G. Thoma, R. Banteli, W.
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2001, 40, 3644–3647; b) G. Thoma, R. O. Duthaler, J. L. Mag-
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c) K. Fukunaga, T. Tsukida, H. Moriyama, H. Kondo, Bioorg.
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Thomas, P. Thakker, J. C. Alvarez, R. T. Camphausen, D.
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a) W. Stahl, U. Sprengard, G. Kretzschmar, H. Kunz, Angew.
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Z.-L. Zheng, C. Perez, L. E. Walker, S. A. DeFrees, F. C. A.
Gaeta, J. Med. Chem. 1994, 37, 3459–3463; c) S. A. DeFrees,
F. C. A. Gaeta, Y.-C. Lin, Y. Ichikawa, C.-H. Wong, J. Am.
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S. Abbas, P. Nikrad, O. Srivastava, C. Foxall, Y. Oda, A. Hase-
gawa, Glycobiology 1993, 3, 633–641; e) D. Tyrrell, P. James,
N. Rao, C. Foxall, S. Abbas, F. Dasgupta, M. Nashed, A.
Hasegawa, M. Kiso, D. Asa, J. Kidd, B. K. Brandley, Proc.
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S. Dolish, A. Aruffo, O. Cecconi, M. P. Bevilacqua, J. Clin.
Invest. 1993, 91, 1157–1166.
[8]
[9]
E- and P-Selectin Inhibition Assays: SLex-BSA (2.3 µg/mL) or
PSGL1/IgG chimera (5 µg/mL) was coated in 96-well plates O/N
at 4 °C, washed with PBS 0.1%BSA 0.05%Tween-20 then blocked
with PBS-BSA 2% for 2 h at 37 °C. After washing, plates were
incubated for 4 h at room temperature with E-sel/µ (1 µg/mL) pre-
complexed with biotinylated goat antihuman IgM (0.5 µg/mL) and
streptavidin-HRPO (0.6 µg/mL) in the presence of the investigated
carbohydrate inhibitors. After extensive washing, selectin binding
was revealed by O-phenylenediamine dihydrochloride (0.67 mg/mL,
Sigma) in the presence of 0.016% H2O2. The reaction was stopped
with H2SO4 (3 ). OD was read at 490 nm.
Supporting Information (see footnote on the first page of this arti-
cle): Copies of the 1H and 13C NMR spectra of all new compounds;
biological details for the selectin inhibition assays.
Acknowledgments
[10]
L. Scheffler, B. Ernst, A. Kapodis, J. L. Magnani, W. T. Wang,
R. Weiseman, T. Peters, Angew. Chem. Int. Ed. Engl. 1995, 34,
1841–1844.
E. E. Simanek, G. J. McGarvey, J. A. Jablonowsky, C.-H.
Wong, Chem. Rev. 1998, 98, 833–862.
We thank the Ministerio de Educación y Ciencia of Spain
(CTQ2004-00649/BQU and CTQ2007-61254/BQU), and the Con-
sejería de Innovación, Ciencia y Empresa de la Junta de Andalucía
(FQM-345 and P07-FQM-03056) for financial support.
[11]
[12]
a) Wong’s fucopeptides: S.-H. Wu, M. Shimazaki, C.-C. Lin,
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Received: February 20, 2008
Published Online: April 24, 2008
2982
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© 2008 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Eur. J. Org. Chem. 2008, 2973–2982