Studies on selectin blockers. 7. Structure-activity relationships of sialyl Lewis X mimetics based on modified ser-glu dipeptides

Article abstract of DOI:10.1021/jm980267x

We have previously found that heterochiral fucodipeptides, L-Ser-D-Glu (3a) and D-Ser-L-Glu (3b), exhibited up to 20-100 times more potency than a sialyl Lewis X (sLe(X), 1) and a 3'sulfated Lewis X analogue (2) toward E- selectin binding and have also proposed, from molecular dynamics calculation, that their strong activities would depend on a possible formation of the type II and/or type II' β-turn of compounds 3a,b (Tsukida, T.; Hiramatsu, Y.; Tsujishita, H.; Kiyoi, T.; Yoshida, M.; Kurokawa, K.; Moriyama, H.; Ohmoto, H.; Wada, Y.; Saito, T.; Kondo, H. J. Med. Chem. 1997, 40, 3534-3541). To clarify our hypothesis, we synthesized several analogues of compounds 3a,b and investigated their structure-activity relationships. As a result, it was indicated that the type II and/or type II' β-turn conformation would be a comparatively tight form and would play important roles in favorable binding to E-selectin. These findings indicate that sLe(X) mimetics with type II and type II' β-turn dipeptides could be a useful methodology for the design of an active selectin blocker.

Full text of DOI:10.1021/jm980267x

J . Med. Chem. 1998, 41, 4279-4287
4279
Stu d ies on Selectin Block er s. 7. Str u ctu r e-Activity Rela tion sh ip s of Sia lyl
Lew is X Mim etics Ba sed on Mod ified Ser -Glu Dip ep tid es
Takahiro Tsukida, Hideki Moriyama, Kiriko Kurokawa, Toshio Achiha, Yoshimasa Inoue, and Hirosato Kondo*
Department of Medicinal Chemistry and Biology, Kanebo, New Drug Discovery Research Laboratories, 1-5-90 Tomobuchi-Cho,
Miyakojima-Ku, Osaka 534, J apan
Received May 1, 1998
We have previously found that heterochiral fucodipeptides, L-Ser-D-Glu (3a ) and D-Ser-L-Glu
(3b), exhibited up to 20-100 times more potency than a sialyl Lewis X (sLeX, 1) and a 3′-
sulfated Lewis X analogue (2) toward E-selectin binding and have also proposed, from molecular
dynamics calculation, that their strong activities would depend on a possible formation of the
type II and/or type II′ â-turn of compounds 3a ,b (Tsukida, T.; Hiramatsu, Y.; Tsujishita, H.;
Kiyoi, T.; Yoshida, M.; Kurokawa, K.; Moriyama, H.; Ohmoto, H.; Wada, Y.; Saito, T.; Kondo,
H. J . Med. Chem. 1997, 40, 3534-3541). To clarify our hypothesis, we synthesized several
analogues of compounds 3a ,b and investigated their structure-activity relationships. As a
result, it was indicated that the type II and/or type II′ â-turn conformation would be a
comparatively tight form and would play important roles in favorable binding to E-selectin.
These findings indicate that sLeX mimetics with type II and type II′ â-turn dipeptides could be
a useful methodology for the design of an active selectin blocker.
Ch a r t 1
In tr od u ction
A multistep series of adhesive events regulate inflam-
matory responses to infection or injury.1 Selectins
mediate the first adhesive step, which is characterized
by rolling of leukocytes on endothelial cells, platelets,
or other leukocytes.2 Therefore, selectins are believed
to be involved in the progression of the clinical mani-
festations of diseases, such as acute and/or chronic
inflammation,3 and have demonstrated a role in the
inflammatory response. Since it has been reported that
all selectins bind to sialyl Lewis X tetrasaccharide (sLeX,
1; Chart 1),4 sLeX-based drug design has attracted the
interest of medicinal chemists. Development of carbo-
hydrate-based therapy is, however, hindered by the
complication that most proteins bind carbohydrates with
weak affinity5 and that the synthesis of complex oli-
gosaccharides is a rather expensive and difficult process.
Such a crucial problem makes sLeX mimetics an attrac-
tive target for useful drug candidates.
We have studied6 the design and synthesis of sugar
mimetics, to develop a methodology for sugar mimetics.
Prediction and understanding of an active conformation
of sugar is very important for the sugar mimetic study.
Our previous studies7 of molecular dynamics simula-
tions of a 3′-sulfo LeX (2; Chart 1)/E-selectin complex
suggested the existence of three essential binding
sites: (1) a fucose for the coordination to calcium, (2) a
branched alkyl chain for the interaction with the
hydrophobic region of E-selectin, and (3) a negatively
charged group for the interaction with the basic residue
of E-selectin. In addition, we have found an active
scaffold, which can conserve the three essential groups
to the desirable positions for the E-selectin binding,
using a computational screening. Namely, we have
proposed that the lactose unit of a 3′-sulfo LeX derivative
(2) could be replaced with a simple dipeptide, D-Ser-L-
Glu (3a ) or L-Ser-D-Glu (3b), characterized by a type II
and/or type II′ â-turn formation (Figure 1).6e There is,
however, little experimental evidence to support that
the type II and/or type II′ â-turn conformation would
be an active scaffold for the sLeX mimics. To clarify the
structure-activity relationships (SAR) of modified Ser-
Glu dipeptides, we next investigated the SAR of com-
pound 3 as illustrated in Figure 2.
In this paper, we describe the synthesis of Ser-Glu
dipeptide derivatives and their in vitro inhibitory activi-
ties against E-, P-, and, L-selectin/sLeX bindings.
Resu lts a n d Discu ssion
Ch em istr y. Our interest was to investigate the SAR
of compounds 3 and to clarify the importance of type II
and/or type II′ â-turn conformation as the active scaffold
for the E-selectin binding. The SAR map of compounds
3 is illustrated in Figure 2.
10.1021/jm980267x CCC: $15.00 © 1998 American Chemical Society
Published on Web 10/02/1998

4280 J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22
Tsukida et al.
F igu r e 1. Successful sLeX mimetics 2 and 3a ,b.
F igu r e 2. SAR map of compounds 3-5.
The construction of dipeptides units (6a -d ) was
carried out according to a previous paper.6e For the
syntheses of compounds 4a -d , the glycosylation of
6a -d with 2,3,4-tri-O-acetyl fucose (7)8 in the presence
of SnCl2/AgOTf as promoters yielded selectively â-gly-
cosides 8a -d in 65-75% yields. The condensation of
8a -d with 1-tetradecylhexadecanoic acid (9) in the
presence of WSC, 1-ethyl-3-[3-(dimethylamino)propyl]-
carbodiimide hydrochloride, and HOBt, 1-hydroxy-1H-
benzotriazole monohydrate, afforded compounds 10a -
d , in 58-62% yields. Finally, compounds 10a -d were
transformed, by removal of the protecting benzyl group
under the general catalytic reduction conditions followed
by treatment with 28% NaOMe in methanol, into the
desired compounds 4a -d , as shown in Scheme 1.
For the synthesis of the intermediates L-13 and D-13
for D-mannose derivatives 5a -d , enantiomeric Ser
units, a L-Ser unit (L-11) and a D-Ser unit (D-11), which
were commercially available, were glycosylated with
1,2,3,4,6-penta-O-acetyl-D-mannose (12) in the presence
of BF3•OEt2 as a promoter to afford the corresponding
R-glycosides (L-13, D-13) stereoselectively. The depro-
tection of the Boc groups of the enantiomeric Glu units
(L-14, D-14), which were synthesized according to a
previous paper,6e with 4 N HCl in 1,4-dioxane at room
temperature followed by coupling with the R-glycosides
(L-13, D-13) in the presence of WSC and HOBt gave
compounds 15a -d in 60-98% yields. The deprotection
of the Fmoc groups of 15a -d with 20% morpholine in
DMF at room temperature afforded the compounds
16a -d , which were next coupled with 9 in the presence
of WSC and HOBt for amide linkage to provide com-
pounds 17a -d in moderate yields. Finally, compounds
17a -d were subjected to hydrogenolysis in the presence
of a catalyst, 10% palladium-carbon, under hydrogen
atmosphere, and then the acetyl groups were removed
by treatment with 28% NaOMe in methanol to give the
desired compounds 5a -d in 78-84% yields, as shown
in Scheme 2.
Compounds 18-21 and 33 were synthesized accord-
ing to a previous paper,6e as shown in Scheme 3. First,
the glycosylation of 6b,e,f with 2,3,4-tri-O-benzylfucose
donor 22 in the presence of SnCl2/AgOTf as promoters
and TMU as a weak base gave mainly R-glycosides 23-
25. The Boc group deprotection of 23-25 in TFA,
followed by the coupling with carboxylic acids 9, 26, and
27 in the presence of WSC and HOBt, afforded com-
pounds 28-32, in 56-76% yields. Next, the compounds
28-32 were transformed, by the removal of the protect-
ing benzyl group under hydrogen atmosphere, into the
desired compounds 3b and 18-21 in 83-94% yields.
Compound 3b was treated with 2 M TMSCH2N2/n-
hexane in methanol-toluene to afford the methyl ester
33 quantitatively.
Biologica l Activities. The method using selectin-
IgG chimeras reported by Foxall et al. was followed.9
To explore the structure-activity relationships of com-
pound 3, we have studied the modification as illustrated

Studies on Selectin Blockers
J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22 4281
Sch em e 1a
a
Conditions: (a) SnCl2, AgOTf, MS (4A)/CH2Cl2, 65-75%; (b) WSC, HOBt, 58-62%; (c) Pd/C/EtOH; (d) 28% NaOMe, 60-95%.
Sch em e 2a
a
Conditions: (a) BF3‚OEt2, CHCl3, 45-63.5%; (b) 4 N HCl/1,4-dioxane; (c) L-13, D-13, WSC, HOBt, 60-98% from L-14, D-14; (d) 20%
morpholine/DMF; (e) 9, WSC, HOBt, 55-67%; (f) Pd/C/98% 1,4-dioxane; (g) 28% NaOMe/MeOH, 78-84% from 17a -d .
in Figure 2: (1) stereochemistry and isostere, (2)
hydrophobic part, (3) negatively charged group, (4)
hydrogen bond for â-turn.
tory activities (IC50 values 3.5 µM for 5a and 16 µM for
5b), and the L-Ser-L-Glu (5c) and the D-Ser-D-Glu (5d ),
however, showed very weak activities (IC50 values of
both 5c,d were >1000 µM). The in vitro profiles of
compounds 5a -d were similar to those of compounds
3a -d (IC50 values 13 µM for 3a , 5.5 µM for 3b, >1000
µM for 3c, and >1000 µM for 3d ). Toward the P- and
L-selectins, all dipeptides 5a -d showed potent blocking
activities, despite their different stereochemical fea-
tures. These findings indicated that the R-D-mannose
could function as an isostere of the R-L-fucose of
compound 3 and that the heterochiral dipeptide back-
bones, the L-Ser-D-Glu and the D-Ser-L-Glu, which were
characterized by type II and type II′ â-turns,6e could play
important roles in favorable binding to E-selectin; in
First, we focused on the stereochemistry and isostere
of the L-fucose of compound 3. The fucose residue was
early recognized as an essential functionality for binding
to selectins,10 and it would have similar calcium-binding
properties to that of the D-mannose in analogous man-
nose-binding proteins.11 Currently, Kogan et al.12 re-
ported that the R-L-fucose unit could be replaced with
an R-D-mannose in a study of selectin blockers. There-
fore, we have also investigated the inhibitory activities
of the D-mannose derivatives 5a -d , against the E-, P-,
and L-selectins, incorporated instead of the L-fucose of
compounds 3a -d . Toward the E-selectin, the L-Ser-D-
Glu (5a ) and the D-Ser-L-Glu (5b) showed potent inhibi-

4282 J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22
Tsukida et al.
Sch em e 3a
a
Conditions: (a) SnCl2, AgOTf, TMU, MS (4A)/CHCl3; (b) TFA/CHCl3; (c) WSC, HOBt, 56-76% from 23, 24, 25; (d) 20% Pd(OH)2/
EtOH, 83-94%; (e) TMSCH2N2/MeOH-toluene, 95%.
Ta ble 1. Blocking Activity of Compounds 4a -d , 5a -d ,
addition, the ligand-binding site to E-selectin would be
18-21, 33, 2, and sLeX
different from those to the P- and L-selectins.
On the other hand, it is known that a sLeX, which is
one of the natural ligands for selectin recognition,4 has
an R-linkage for the L-fucosyl bond. Recently, Wong et
al.13 reported that a â-fucosyl dipeptide showed inhibi-
tory activity as well as the corresponding R-anomer
against the E-selectin binding in a study of sLeX
IC50, µM
mimetics. Thus, to study the stereochemistry-activity
relationships concerning the L-fucose of compound 3, we
synthesized the corresponding â-anomers 4a -d to the
R-anomers 3a -d and evaluated their inhibitory activi-
ties against E-, P-, and L-selectin bindings as shown in
Table 1. As a result, regardless of the difference of the
chirality of the dipeptide backbones, all compounds
4a -d had very weak inhibitory activities (IC50 > 1000
µM) toward E-selectin binding. Namely, it was found
that the R-configuration of the L-fucose of compound 3
was very important for E-selectin recognition. Our
finding indicates that the compound 4 described here
would have a comparatively tight conformation, such
as the type II and type II′ â-turns, compared to the
â-fucosyl dipeptide reported by Wong et al. Toward the
P- and L-selectins, all of the dipeptides 4a -d showed
potent blocking activities.
Next, we investigated the modification of the hydro-
phobic part of compound 3b, which was one of the most
potent blockers against E-selectin. Compound 18,
including a shorter branched alkyl chain, 11-carbon
length, compared to compound 3b was 20-30 times less
active than compound 3b toward E- and P-selectins. In
addition, compound 19 having a single alkyl chain
showed significantly weak inhibitory activity to all
selectins. To clarify the importance of the branched
alkyl chain of compounds 5a ,b toward the inhibitory
activity, we have studied the active forms of compounds
compd
*1
*2
E-selectin
P-selectin
L-selectin
4a
4b
4c
4d
5a
5b
5c
5d
18
19
20
21
33
2
L
D
L
D
L
L
D
D
L
L
D
L
L
L
L
>100
>100
>1000
>1000
3.5
0.3
0.3
0.6
0.3
0.45
0.42
0.68
0.38
15
0.8
1.6
1.8
D
L
0.7
4
D
L
16
3.1
>1000
>1000
160
4.2
D
D
D
D
D
4.4
12
>500
>500
>500
>500
280
250-500
200
3.8
3.6
1.6
2.4
37
100
>1000
>500
30
sLeX
600
>1000
5a ,b, like a micellar species, in solution. Namely, we
have measured the UV absorbance to investigate light
scattering changes on the concentration with varying
concentrations of compounds 5a ,b. As a result, the
critical micelle concentration (CMC) could not be seen
at IC50 values 3.5 and 16 µM, respectively. These
findings demonstrate that the branched alkyl chains
with a moderate carbon length would be necessary for
the strong bindings to all selectins.
Current sLeX mimetic studies show that the sialic acid
residue of sLeX could be easily replaced by negatively
charged groups, such as carboxylic acid,14 sulfuric acid,15
and phosphoric acid,6a and the negatively charged group

Studies on Selectin Blockers
J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22 4283
to cause 50% inhibition of the ligand binding to each of the
selectins by the compound. The number of replicates is 2.
was essential for E-, P-, and L-selectins. We have also
reported that the negatively charged group, a carboxy-
late of the Glu unit of 3a ,b, would be important for the
ionic interaction with the basic residue Lys 111 in
E-selectin by molecular dynamics simulation.6e To
clarify this point, we have synthesized the compound
33 including a methyl carboxylate masked with a
methyl group. As expected, compound 33 had very
weak activity (IC50 > 500 µM) toward E-selectin. On
the other hand, compound 20 containing a L-Asp unit
instead of a L-Glu unit had markedly reduced inhibitory
activity (IC50 > 500 µM) against E-selectin. Namely,
this result indicates that there is a moderate position
of the negatively charged group necessary for the
E-selectin binding.
Finally, to clarify the importance of the possible
â-turn conformation of compounds 3a ,b, we synthesized
compound 21 lacking an amide proton necessary for the
formation of â-turn and investigated the inhibitory
activity against E-selectin. It was of interest that the
inhibitory activity of compound 21 toward the E-selectin
was dramatically decreased, IC50 > 500 µM.
Gen er a l P r oced u r e for th e P r ep a r a tion of â-Glyco-
sid es 8a -d by Glycosyla tion of 6a -d w ith 2,3,4-Tr i-O-
a cetylfu cose Don or 7: [O-(2,3,4-Tr i-O-a cetyl-â-L-fu cop y-
r an osyl)-D-ser yl]-L-glu tam ic Acid 1-Meth ylam ide 5-Ben zyl
Ester (8b). A mixture of molecular sieves (4 Å, 1.0 g), AgOTf
(1.41 g, 5.48 mM), and SnCl2 (1.04 g, 5.48 mM) in CH2Cl2 (10
mL) was stirred for 0.5 h under argon atmosphere at room
temperature and cooled to -20 °C. 2,3,4-Tri-O-acetyl-L-fucosyl
fluoride (7) (1.20 g, 4.11 mM) in CH2Cl2 (3 mL) and compound
6b (1.20 g, 2.74 mM) in CH2Cl2 (5 mL) were added to the
mixture successively and stirred for 21 h while gradually
returning to room temperature. The precipitate was filtered
off, and the filtrate was concentrated in vacuo. The resulting
residue was purified by thin-layer chromatography developing
with 9:1 CHCl3/methanol to give 8b (1.24 g, 74.3%) as a
syrup: 1H NMR (CDCl3) δ 1.2 (d, 3H, J ) 6.4 Hz), 1.99 (s,
9H), 2.07 (s, 3H), 2.13 (s, 3H), 2.15-2.4 (m, 3H), 2.4-2.6 (m,
2H), 2.8 (d, 3H, J ) 4.7 Hz), 3.49-3.62 (m, 1H), 3.7 (dd, 1H,
J ) 3.8, 13.6 Hz), 3.76-3.88 (m, 1H), 4.02-4.16 (m, 1H), 4.95-
5.20 (m, 3H), 5.23 (d, 1H, J ) 2.6 Hz), 6.78 (d, 1H, J ) 4.7
Hz), 7.35 (s, 5H), 7.87 (d, 1H, J ) 8.2 Hz). Anal. (C28H39N3O12
C, H, N.
)
[O-(2,3,4-Tr i-O-a cet yl-â-L-fu cop yr a n osyl)-L-ser yl]-D-
glu ta m ic a cid 1-m eth yla m id e 5-ben zyl ester (8a ): yield
65.6% as a syrup; 1H NMR (CDCl3) δ 1.20 (d, 3H, J ) 6.3 Hz),
1.98 (s, 9H), 2.03 (s, 3H), 2.08 (t, 1H, J ) 2.7 Hz), 2.16 (s, 3H),
2.15-2.35 (m, 1H), 2.38-2.57 (m, 2H), 2.79 (d, 3H, J ) 4.7
Hz), 3.56 (t, 1H, J ) 5.1 Hz), 3.72-3.95 (m, 2H), 3.99 (dd, 1H,
J ) 3.8, 9.4 Hz), 4.44 (d, 1H, J ) 7.6 Hz), 4.39-4.52 (m, 1H),
5.0 (dd, 1H, J ) 3.3, 10.5 Hz), 5.04-5.18 (m, 3H), 5.23 (d, 1H,
J ) 3.3 Hz), 6.67 (d, 1H, J ) 4.5 Hz), 7.28-7.40 (m, 5H), 8.0
(d, 1H, J ) 8.6 Hz). Anal. (C28H39N3O12) C, H, N.
In conclusion, we have investigated the SAR of
compound 3 found currently using a computational
screening, and we have found that a Ser-Glu dipeptide
backbone, which is characterized by a type II and/or
type II′ â-turn, could be a useful template for the design
of sLeX mimetics.
Exp er im en ta l Section
In h ibition Assa y of Selectin -sLeX Bin d in g. The con-
struction of the selectin-immunoglobulin was carried out
according to a previous paper.16
[O-(2,3,4-Tr i-O-a ce t yl-â-L-fu cop yr a n osyl)-L-se r yl]-L-
glu ta m ic a cid 1-m eth yla m id e 5-ben zyl ester (8c): crude
8c was used without further purification.
A solution of sLeX pentasaccharide ceramide analogue in a
1:1 mixture of methanol and distilled water was pipetted into
microtiter plate wells (96 wells; Falcon PRO-BIND) at 100
pmol/50 µL/well and was adsorbed by evaporating the solvent.
The wells were washed twice with distilled water, blocked with
5% BSA (bovine serum albumin)-1 mM CaCl2/50 mM imida-
zole buffer (pH 7.2) for 1 h at room temperature, and washed
three times with 50 mM imidazole buffer (pH 7.2).
[O-(2,3,4-Tr i-O-a cet yl-â-L-fu cop yr a n osyl)-D-ser yl]-D-
glu ta m ic a cid 1-m eth yla m id e 5-ben zyl ester (8d ): yield
68.1% as a syrup; 1H NMR (CDCl3) δ 1.18 (d, 3H, J ) 6.4 Hz),
1.99 (s, 9H), 2.05 (s, 3H), 2.12 (s, 3H), 2.35-2.55 (m, 2H), 2.76
(d, 3H, J ) 4.6 Hz), 3.5-3.66 (m, 1H), 3.74-3.96 (m, 3H), 4.37-
4.55 (m, 1H), 4.51 (d, 1H, J ) 7.6 Hz), 5.02 (dd, 1H, J ) 3.4,
10.4 Hz), 5.10 (s, 2H), 5.21 (d, 1H, J ) 3.2 Hz), 7.08 (q, 1H, J
) 4.8 Hz), 7.25-7.38 (m, 5H), 7.93 (d, 1H, J ) 8.4 Hz). Anal.
(C28H39N3O12) C, H, N.
Gen er a l P r oced u r e for th e P r ep a r a tion of 10a -d :
[N-(2-Tetr a d ecylh exa d eca n oyl)-O-(2,3,4-tr i-O-a cetyl-â-L-
fu cop yr a n osyl)-D-ser yl]-L-glu ta m ic Acid 1-Meth yla m id e
5-Ben zyl Ester (10b). To a solution of 8c (1.20 g, 1.97 mM)
in DMF (50 mL) was added 2-tetradecylhexadecanoic acid (0.89
g, 1.97 mM), and the mixture was dissolved by heating and
then cooled to room temperature. WSC (0.57 g, 2.96 mM) and
HOBt (0.45 mg, 2.96 mM) were added to the solution. After
the mixture stirred for 16.5 h, AcOEt (120 mL) was added to
the solution, and the mixture was washed with 1 N HCl,
saturated sodium hydrogen carbonate, and brine successively,
dried (MgSO4), and concentrated. The resulting residue was
purified by thin-layer chromatography developing with 20:1
CHCl3/methanol to give 10b (1.20 g, 58.3%) as a syrup: 1H
NMR (CDCl3) δ 0.8-0.95 (m, 6H), 0.96-1.68 (m, 60H), 1.99
(s, 3H), 2.04 (s, 3H), 2.10 (s, 3H), 2.40-2.64 (m, 2H), 2.75 (d,
3H, J ) 4.7 Hz), 3.78 (dd, 1H, J ) 3.3, 11.0 Hz), 3.83-4.05
(m, 2H), 4.4-4.67 (m, 2H), 4.59 (d, 1H, J ) 7.7 Hz), 5.02 (dd,
1H, J ) 3.3, 10.4 Hz), 5.06-5.2 (m, 3H), 5.25 (d, 1H, J ) 3.2
Hz), 6.62 (d, 1H, J ) 8.2 Hz), 6.7 (d, 1H, J ) 4.8 Hz), 7.20-
7.42 (m, 5H). Anal. (C58H97N3O13) C, H, N.
Separately, a 1:1 volumetric mixture of a 1:500 dilution in
1% BSA-1 mM CaCl2/50 mM imidazole buffer (pH 7.2) of
biotinylated goat F(ab′)2 anti-human IgG(g)/streptavidin-
alkaline phosphatase (Zymed Lab Inc.) and a selectin-immu-
noglobulin fusion protein (selectin-Ig) was incubated at room
temperature for 30 min to form a complex. The test com-
pounds were dissolved in DMSO at 10 mM and finally diluted
by 1 mM CaCl2/50 mM imidazole buffer (pH 7.2) to final
concentrations at 100, 50, 25, 12.5, 6.25, 3.13, 1.56, and 0.78
µM, respectively. Reactant solutions were prepared by incu-
bating 30 µL of this solution at each concentration with 30 µL
of the above complex solution for 30 min at room temperature.
This reactant solution was then added to the above microtiter
wells at 50 µL/well and incubated at 37 °C for 45 min. The
wells were washed three times with 50 mM imidazole buffer
(pH 7.2) and distilled water, respectively, followed by addition
of p-nitrophenyl phosphate (1 mg/mL) and 0.01% MgCl2 in 1
M diethanolamine (pH 9.8) at 50 µL/well. The reactant
mixture was developed for 120 min at room temperature, and
absorbance at 405 nm was measured. Percent binding was
calculated by the following equation:
% binding ) (X - C/A - C) × 100
[N-(2-Tetr a d ecylh exa d eca n oyl)-O-(2,3,4-tr i-O-a cetyl-â-
L-fu cop yr a n osyl)-L-ser yl]-D-glu ta m ic a cid 1-m eth yla m id e
5-ben zyl ester (10a ): yield 61.8% as a syrup; 1H NMR (CDCl3)
δ 0.8-1.00 (m, 6H), 1.19 (d, 3H, J ) 6.4 Hz), 1.20-1.38 (m,
52H), 1.38-1.66 (m, 2H), 1.70 (s, 2H), 1.98 (s, 3H), 2.07 (s,
3H), 2.13 (s, 3H), 2.17-2.40 (m, 2H), 2.79 (d, 3H, J ) 4.7 Hz),
3.74-3.92 (m, 2H), 3.99 (dd, 1H, J ) 4.4, 9.4 Hz), 4.37-4.56
wherein X is the absorbance of wells containing the test
compounds at each concentration, C is the absorbance of wells
not containing the selectin-Ig and test compounds, and A is
the absorbance of control wells not containing the test com-
pounds. The results of inhibitory activities are presented in
Table 1 as IC50 values, the concentrations of blockers necessary

4284 J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22
Tsukida et al.
(m, 2H), 4.49 (d, 1H, J ) 7.3 Hz), 4.95-5.2 (m, 4H), 5.2-5.3
(m, 1H), 6.44 (d, 1H, J ) 5.6 Hz), 6.96 (d, 1H, J ) 7.9 Hz),
7.35 (s, 5H). Anal. (C58H97N3O13) C, H, N.
(dd, 1H, J ) 4.6, 9.8 Hz), 4.07 (d, 1H, J ) 7.3 Hz), 4.12-4.3
(m, 1H), 4.3-4.66 (m, 2H), 4.67-4.8 (m, 1H), 7.56 (d, 1H, J )
4.7 Hz), 7.92 (d, 1H, J ) 7.4 Hz), 7.98 (d, 1H, J ) 8.2 Hz),
12.1 (s, 1H); MS spectrum 828 (M + H)+. Anal. (C45H85N3O10
‚
[N-(2-Tetr a d ecylh exa d eca n oyl)-O-(2,3,4-tr i-O-a cetyl-â-
L-fu cop yr a n osyl)-L-ser yl]-L-glu ta m ic a cid 1-m eth yla m id e
5-ben zyl ester (10c): yield 60% as a white crystal; mp 102-
104 °C; 1H NMR (CDCl3) δ 0.85-0.9 (m, 6H), 1.19-1.3 (m,
55H), 1.38-1.57 (m, 2H), 1.70 (s, 2H), 1.99 (s, 3H), 2.08 (s,
3H), 2.11 (s, 3H), 2.51-2.59 (m, 2H), 2.84 (d, 3H, J ) 4.7 Hz),
3.78-3.85 (m, 2H), 3.94 (dd, 1H, J ) 4.6, 9.0 Hz), 4.41-4.54
(m, 2H), 4.47 (d, 1H, J ) 7.2 Hz), 4.96-5.1 (m, 2H), 5.12 (s,
2H), 5.24-5.29 (m, 1H), 6.5 (d, 1H, J ) 6.3 Hz), 6.72 (d, 1H,
J ) 4.7 Hz), 6.94 (d, 1H, J ) 8.3 Hz), 7.34 (s, 5H). Anal.
(C58H97N3O13) C, H, N.
1.5H2O) C, H, N.
Gen er a l P r oced u r e for th e P r ep a r a tion of L-13 a n d
D-13 by Glycosyla tion of L-12 a n d D-12 w ith 1,2,3,4,6-
P en ta -O-a cetyl-D-m a n n op yr a n ose Don or 11: N-(9-F lu o-
r en ylm et h yloxyca r b on yl)-O-(2,3,4,6-t et r a -O-a cet yl-r-D-
m a n n op yr a n osyl)-L-ser in e (L-13). To a mixture of N-(9-
fluorenylmethyloxycarbonyl)-D-serine (0.9 g, 2.75 mM) and
1,2,3,4,6-penta-O-acetyl-D-mannopyranose (0.9 g, 3.64 mM) in
CHCl3 (36 mL) was added BF3‚OEt2 (2.6 mL), and the mixture
was stirred for 21 h under nitrogen at room temperature.
Concentrated, the residue was dissolved in AcOEt (90 mL),
washed with water, dried (MgSO4), and concentrated in vacuo.
The resulting residue was purified by chromatography (SiO2,
gradient elution CHCl3/methanol, 30:1 to 20:1) to give L-13
(1.15 g, 63.5%) as a syrup: 1H NMR (CDCl3) δ 1.96, 2.03, 2.16
(3s, 12H), 3.9-4.2 (m, 3H), 4.2-4.3 (m, 2H), 4.3-4.8 (m, 3H),
4.88 (bs, 1H), 5.1-5.4 (m, 3H), 6.1 (bs, 1H), 7.2-7.5 (m, 5H),
7.8 (d, 2H, J ) 7.4 Hz). Anal. (C32H35NO14) C, H, N.
N-(9-F lu or en ylm eth yloxyca r bon yl)-O-(2,3,4,6-tetr a -O-
a cetyl-r-D-m a n n op yr a n osyl)-D-ser in e (D-13): yield 45.0%
as a syrup; 1H NMR (CDCl3) δ 1.98, 2.09, 2.14 (3s, 12H), 3.65-
3.85 (m, 1H), 3.98 (bs, 1H), 4.05-4.3 (m, 4H), 4.3-4.7 (m, 3H),
4.83 (bs, 1H), 5.15-5.4 (m, 3H), 5.9 (bs, 1H), 7.2-7.45 (m, 5H),
7.61 (d, 2H, J ) 6.4 Hz), 7.75 (d, 2H, J ) 7.4 Hz). Anal.
(C32H35NO14) C, H, N.
Gen er a l P r oced u r e for th e P r ep a r a tion of 15a -d :
[N-(9-F lu or en ylm et h yloxyca r b on yl)-O-(2,3,4,6-t et r a -O-
a cetyl-r-D-m a n n op yr a n osyl)-D-ser yl]-L-glu ta m ic Acid 1-
Meth yla m id e 5-Ben zyl Ester (15b). To the L-14 (575 mg,
1.64 mM) was added 4 N HCl/1,4-dioxane (3 mL), and the
mixture was stirred for 15 min at room temperature. The
mixture was concentrated, and the resulting residue and D-13
(0.9 g, 1.37 mM) were dissolved in DMF (25 mL), added to
Et3N (180 mg, 1.78 mM), and then cooled to 0 °C. WSC (315
mg, 1.64 mM) and HOBt (252 mg, 1.64 mM) were added to
the solution, and this was stirred for 18 h at room temperature.
AcOEt (120 mL) was added, and the mixture was washed with
1 N HCl, saturated sodium hydrogen carbonate, and brine
successively, dried (MgSO4), and concentrated. The resulting
residue was purified by thin-layer chromatography developing
with 20:1 CHCl3/methanol to give 15b (767 mg, 62.9%) as a
white crystal: 1H NMR (CDCl3) δ 1.98, 2.00, 2.10, 2.15 (4s,
12H), 1.9-2.7 (m, 4H), 2.78 (d, 3H, J ) 4.8 Hz), 3.54 (dd, 1H,
J ) 4.8, 10.0 Hz), 3.97 (bs, 1H), 4.1-4.3 (m, 4H), 4.3-4.5 (m,
4H), 4.8 (s, 1H), 5.06 (d, 1H, J ) 12.3 Hz), 5.12 (d, 1H, J )
12.3 Hz), 5.15-5.35 (m, 3H), 5.86 (d, 1H, J ) 7.4 Hz), 6.36 (q,
1H, J ) 4.8 Hz), 7.2-7.45 (m, 9H), 7.62 (d, 2H, J ) 7.2 Hz),
7.76 (d, 2H, J ) 7.3 Hz). Anal. (C45H51N3O16) C, H, N.
[N-(2-Tetr a d ecylh exa d eca n oyl)-O-(2,3,4-tr i-O-a cetyl-â-
L-fu cop yr a n osyl)-D-ser yl]-D-glu t a m ic a cid 1-m et h yl-
a m id e 5-ben zyl ester (10d ): yield 59.6% as a white crystal;
mp 103-105 °C; 1H NMR (CDCl3) δ 0.80-0.95 (m, 6H), 1.05-
1.75 (m, 55H), 1.99 (s, 3H), 2.05 (s, 3H), 2.11 (s, 3H), 2.2-2.42
(m, 1H), 2.43-2.63 (m, 2H), 2.74-2.8 (m, 1H), 2.84 (d, 3H, J
) 4.7 Hz), 3.79 (dd, 1H, J ) 8.5, 11.2 Hz), 3.86-3.97 (m, 1H),
3.98-4.08 (m, 1H), 4.37-4.52 (m, 1H), 4.50-4.73 (m, 1H), 4.63
(d, 1H, J ) 7.7 Hz), 4.99 (dd, 1H, J ) 3.2, 10.4 Hz), 5.04-5.07
(m, 3H), 5.25 (d, 1H, J ) 2.7 Hz), 6.46 (d, 1H, J ) 5.9 Hz),
6.65 (d, 1H, J )4.8 Hz), 7.27-7.42 (m, 5H). Anal. (C58H97N3O13
C, H, N.
)
Gen er al P r ocedu r e for th e P r epar ation of 4a-d: [N-(2-
Tetr a decylh exa deca n oyl)-O-(â-L-fu cop yr a n osyl)-D-ser yl]-
L-glu ta m ic Acid 1-Meth yla m id e (4b). To a solution of 10b
(0.25 g, 0.239 mM) in ethanol (30 mL) was added Pd/C (50
mg), and the mixture was stirred for 2 h under hydrogen
atmosphere. The precipitate was filtered off, and the filtrate
was concentrated in vacuo. The residue was dissolved in
methanol (20 mL), 28% NaOMe/methanol (74 mg, 0.478 mM)
was added to the solution, and this was stirred for 0.5 h at
room temperature. DOWEX 50W-X8 (10 g) was added to the
mixture, and this was stirred for 3 min and then added to
CHCl3 (10 mL). The precipitate was filtered off; the filtrate
was concentrated in vacuo to give the desired compound 4b
(0.13 g, 64.6%) as a white powder: [R]D -4° (c ) 0.1, MeOH);
1
mp gradually melted at 163 °C; H NMR (DMSO-d6) δ 0.8-
0.9 (m, 6H), 0.95-1.55 (m, 52H), 1.6-1.8 (m, 1H), 1.8-2.08
(m, 1H), 2.16-2.36 (m, 3H), 2.58 (d, 3H, J ) 4.3 Hz), 3.42-
3.6 (m, 3H), 3.82-4.0 (m, 1H), 4.08 (d, 1H, J ) 6.8 Hz), 4.1-
4.28 (m, 1H), 4.27-4.5 (m, 2H), 4.69 (s, 1H), 4.83 (s, 1H), 7.8
(d, 1H, J ) 4.6 Hz), 7.89 (d, 1H, J ) 8.0 Hz), 8.0 (d, 1H, J )
7.3 Hz), 12.0 (s, 1H); MS spectrum 828 (M + H)+. Anal.
(C45H85N3O10‚1.5H2O) C, H, N.
[N-(2-Tetr a d ecylh exa d eca n oyl)-O-(â-L-fu cop yr a n osyl)-
L-ser yl]-D-glu ta m ic a cid 1-m eth yla m id e (4a ): yield 94.6%
as a white powder; [R]D +5° (c ) 0.1, MeOH); mp 198-199 °C;
1H NMR (DMSO-d6) δ 0.7-0.93 (m, 6H), 0.95-1.55 (m, 55H),
1.55-1.8 (m, 1H), 1.8-2.04 (m, 2H), 2.06-2.3 (m, 3H), 2.57
(d, 3H, J ) 4.3 Hz), 3.4-3.68 (m, 2H), 3.7-3.89 (m, 1H), 4.0-
4.24 (m, 2H), 4.13 (d, 1H, J ) 6.9 Hz), 4.23-4.47 (m, 2H), 4.68
(s, 1H), 7.77 (d, 1H, J ) 4.6 Hz), 7.99 (d, 1H, J ) 6.7 Hz), 8.02
(d, 1H, J ) 8.0 Hz), 12.1 (s, 1H); MS spectrum 828 (M + H)+.
Anal. (C45H85N3O10‚1.5H2O) C, H, N.
[N-(9-F lu or en ylm eth yloxyca r bon yl)-O-(2,3,4,6-tetr a -O-
a cetyl-r-D-m a n n op yr a n osyl)-L-ser yl]-D-glu ta m ic a cid 1-
1
m eth yla m id e 5-ben zyl ester (15a ): yield 97.5%; H NMR
(CDCl3) δ 1.97, 2.00, 2.08, 2.15 (4s, 12H), 1.9-2.7 (m, 4H), 2.77
(d, 3H, J ) 4.8 Hz), 3.9 (bs, 1H), 4.1-4.5 (m, 9H), 4.83 (bs,
1H), 5.0-5.2 (m, 3H), 5.3-5.4 (m, 3H), 5.8 (bs, 1H), 6.3 (bs,
1H), 7.3-7.45 (m, 9H), 7.61 (d, 2H, J ) 7.2 Hz), 7.76 (d, 2H,
J ) 7.1 Hz), 8.00 (bs, 1H). Anal. (C45H51N3O16) C, H, N.
[N-(9-F lu or en ylm eth yloxyca r bon yl)-O-(2,3,4,6-tetr a -O-
a cetyl-r-D-m a n n op yr a n osyl)-L-ser yl]-L-glu ta m ic a cid 1-
m eth yla m id e 5-ben zyl ester (15c): yield 60.7% as a crystal;
1H NMR (CDCl3) δ 1.97, 2.00, 2.08, 2.15 (4s, 12H), 1.9-2.7
(m, 4H), 2.77 (d, 3H, J ) 4.8 Hz), 3.9-4.0 (m, 1H), 4.1-4.5
(m, 6H), 4.8 (bs, 1H), 5.0-5. (m, 2H), 5.2-5.4 (m, 3H), 5.7
(bs, 1H), 6.3 (bs, 1H), 7.2-7.5 (m, 9H), 7.61 (d, 2H, J ) 7.2
Hz), 7.76 (d, 2H, J ) 7.4 Hz). Anal. (C45H51N3O16) C, H, N.
[N-(2-Tetr a d ecylh exa d eca n oyl)-O-(â-L-fu cop yr a n osyl)-
L-ser yl]-L-glu ta m ic a cid 1-m eth yla m id e (4c): yield 60.9%
as a white powder; [R]D -9° (c ) 0.1, MeOH); mp gradually
1
melted at 160 °C; H NMR (DMSO-d6) δ 0.82-0.87 (m, 6H),
1.13 (d, 3H, J ) 6.4 Hz), 1.18-1.41 (m, 52H), 1.65-1.73 (m,
1H), 1.83-1.93 (m, 1H), 2.12-2.19 (m, 3H), 2.53 (d, 3H, J )
4.5 Hz), 3.34-3.58 (m, 3H), 3.68-3.75 (m, 1H), 4.06-4.15 (m,
2H), 4.38-4.48 (m, 2H), 4.70 (s, 1H), 7.54 (d, 1H, J ) 4.7 Hz),
7.87-7.95 (m, 2H), 12.01 (bs, 1H); MS spectrum 828 (M + H)+.
Anal. (C45H85N3O10‚2H2O) C, H, N.
[N-(2-Tetr a d ecylh exa d eca n oyl)-O-(-L-fu cop yr a n osyl)-
D-ser yl]-D-glu ta m ic a cid 1-m eth yla m id e (4d ): yield 65.5%
as a white powder; [R]D +6° (c ) 0.1, MeOH); mp gradually
melted at 147 °C; 1H NMR (DMSO-d6) δ 0.8-0.9 (m, 6H),
0.95-1.6 (m, 55H), 1.6-1.82 (m, 1H), 1.82-2.07 (m, 1H), 2.08-
2.36 (m, 3H), 2.59 (d, 3H, J ) 4.5 Hz), 3.46-3.85 (m, 3H), 3.94
[N-(9-F lu or en ylm eth yloxyca r bon yl)-O-(2,3,4,6-tetr a -O-
a cetyl-r-D-m a n n op yr a n osyl)-D-ser yl]-D-glu ta m ic a cid 1-
m eth yla m id e 5-ben zyl ester (15d ): yield 73.5% as a pale-
yellow crystal; H NMR (CDCl3) δ 1.95, 1.97, 2.10, 2.16 (4s,
12H), 2.05-2.2 (m, 2H), 2.5-2.7 (m, 2H), 2.77 (d, 3H, J ) 4.8
Hz), 3.53 (dd, 1H, J ) 4.1, 8.9 Hz), 3.94 (bs, 1H), 4.1-4.3 (m,
1

Studies on Selectin Blockers
J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22 4285
4H), 4.3-4.5 (m, 3H), 4.84 (s, 1H), 5.04 (d, 1H, J ) 12.2 Hz),
5.11 (d, 1H, J ) 12.2 Hz), 5.23-5.3 (m, 2H), 5.72 (bs, 1H),
6.36 (bs, 1H), 7.1-7.4 (m, 9H), 7.62 (d, 2H, J ) 6.6 Hz), 7.76
(d, 2H, J ) 7.7 Hz). Anal. (C45H51N3O16) C, H, N.
139 °C; 1H NMR (CDCl3) δ 0.8-0.95 (m, 6H), 1.1-1.5 (m, 52H),
1.95, 2.02, 2.09, 2.13 (4s, 12H), 2.4-2.6 (m, 2H), 2.78 (d, 3H,
J ) 4.8 Hz), 3.8-3.9 (m, 2H), 3.98 (bs, 1H), 4.11 (dd, 1H, 2.4,
12.3 Hz), 4.27 (dd, 1H, J ) 5.4, 12.3 Hz), 4.35-4.5 (m, 1H),
4.5-4.65 (m, 1H), 4.86 (d, 1H, J ) 1.2 Hz), 5.09 (d, 1H, J )
12.3 Hz), 5.15 (d, 1H, J ) 12.3 Hz), 5.2-5.32 (m, 3H), 6.25-
6.45 (m, 2H), 7.3-7.4 (m, 5H). Anal. (C60H99N3O15) C, H, N.
[N-(2-Tetr adecylh exadecan oyl)-O-(2,3,4,6-tetr a-O-acetyl-
r-D-m a n n op yr a n osyl)-D-ser yl]-D-glu ta m ic a cid 1-m eth yl-
a m id e 5-ben zyl ester (17d ): yield 67.0% as a pale-yellow
crystal; 1H NMR (CDCl3) δ 0.85-0.9 (m, 6H), 1.0-1.7 (m, 52H),
1.93, 1.99, 2.11, 2.15 (4s, 12H), 2.05-2.25 (m, 2H), 2.46-2.65
(m, 2H), 2.78 (d, 3H, J ) 4.6 Hz), 3.52 (dd, 1H, J ) 5.1, 9.4
Hz), 3.97 (bs, 1H), 4.1-4.3 (m, 3H), 4.4-4.55 (m, 1H), 4.62
(bs, 1H), 4.83 (s, 1H), 5.08 (d, 1H, J ) 12.3 Hz), 5.14 (d, 1H, J
) 12.3 Hz), 5.2-5.35 (m, 3H), 6.3-6.5 (m, 2H), 7.3-7.4 (m,
5H), 7.53 (bs, 1H). Anal. (C60H99N3O15) C, H, N.
Gen er al P r ocedu r e for th e P r epar ation of 5a-d: [N-(2-
Tetr a d ecylh exa d eca n oyl)-r-D-m a n n op yr a n osyl)-D-ser yl]-
L-glu ta m ic Acid 1-Meth yla m id e (5b). To a solution of 17b
(347 mg, 0.315 mM) in 98% 1,4-dioxane (30 mL) was added
Pd/C (350 mg), and the mixture was stirred for 1.5 h under
hydrogen atmosphere. The precipitate was filtered off, and
the filtrate was concentrated in vacuo. The residue was
dissolved in methanol (35 mL), 28% NaOMe/methanol (122
mg) was added to the solution, and this was stirred for 1.5 h
at room temperature. DOWEX (1.0 g) was added to the
mixture, and this was stirred for 3 min. The precipitate was
filtered off, and the filtrate was concentrated under reduced
pressure to give the desired compound 5b (223 mg, 79.0%) as
a white powder: [R]D 23° (c ) 0.1, MeOH); mp 203-206 °C;
1H NMR (DMSO-d6) δ 0.75-1.0 (m, 6H), 1.05-1.55 (m, 52H),
1.6-1.8 (m, 1H), 1.8-2.05 (m, 1H), 2.56 (d, 3H, J ) 4.5 Hz),
3.2-3.8 (m, 8H), 4.1-4.25 (m, 1H), 4.25-4.4 (m, 1H), 4.4-4.5
(m, 1H), 4.5-4.7 (m, 2H), 7.76 (q, 1H, J ) 4.7 Hz), 8.02 (d,
1H, J ) 7.7 Hz), 12.0 (s, 1H). Anal. (C45H85N3O11) C, H, N.
[N-(2-Tetr a d ecylh exa d eca n oyl)-r-D-m a n n op yr a n osyl)-
L-ser yl]-D-glu ta m ic a cid 1-m eth yla m id e (5a ): yield 82.9%
as a white crystal; mp 162-168 °C; 1H NMR (DMSO-d6) δ
0.75-0.95 (m, 6H), 1.1-1.6 (m, 52H), 1.6-1.8 (m, 1H), 1.85-
2.05 (m, 1H), 2.1-2.35 (m, 3H), 2.57 (d, 3H, J ) 4.5 Hz), 3.35-
3.8 (m, 6H), 4.1-4.25 (m, 1H), 4.35-4.5 (m, 1H), 4.65 (d, 1H,
J ) 1.4 Hz), 7.75 (q, 1H, J ) 4.5 Hz), 7.9-8.15 (m, 1H). Anal.
(C45H85N3O11) C, H, N.
Gen er a l P r oced u r e for th e P r ep a r a tion of 16a -d by
th e F m oc Gr ou p Dep r otection of 15a -d : [O-(2,3,4,6-
Tetr a -O-a cetyl-r-D-m a n n op yr a n osyl)-D-ser yl]-L-glu ta m -
ic Acid 1-Meth yla m id e 5-Ben zyl Ester (16b). To 15b (0.5
g, 0.56 mM) was added 20% morpholine/DMF (12 mL), and
the solution was stirred for 1.5 h at room temperature. AcOEt
(100 mL) was added, and this was washed with water twice,
dried (MgSO4), and concentrated in vacuo. The resulting
residue was purified by thin-layer chromatography developing
with 10:1 CHCl3/methanol to give 16b (347 mg, 92.5%) as a
syrup: 1H NMR (CDCl3) δ 1.97, 2.03, 2.11, 2.14 (4s, 12H), 1.9-
2.6 (m, 4H), 2.77 (d, 3H, J ) 4.8 Hz), 3.45-3.55 (m, 1H), 3.62
(dd, 1H, J ) 3.6, 9.5 Hz), 3.9-4.07 (m, 2H), 4.1-4.2 (m, 1H),
4.25 (dd, 1H, 5.2, 12.3 Hz), 5.15-5.35 (m, 3H), 5.86 (d, 1H, J
) 7.4 Hz), 6.58 (q, 1H, J ) 4.7 Hz), 7.2-7.4 (m, 5H), 8.03 (d,
1H, J ) 8.3 Hz). Anal. (C30H41N3O14) C, H, N.
[O-(2,3,4,6-Tetr a-O-acetyl-r-D-m an n opyr an osyl)-L-ser yl]-
D-glu t a m ic a cid 1-m et h yla m id e 5-b en zyl est er (16a ):
crude 16a was used without further purification.
[O-(2,3,4,6-Tetr a-O-acetyl-r-D-m an n opyr an osyl)-L-ser yl]-
L-glu ta m ic a cid 1-m eth yla m id e 5-ben zyl ester (16c): yield
70.1% as a syrup; 1H NMR (CDCl3) δ 1.98, 2.03, 2.10, 2.14
(4s, 12H), 2.0-2.25 (m, 2H), 2.25-2.65 (m, 2H), 2.78 (d, 3H, J
) 4.7 Hz), 3.4-3.7 (m, 2H), 3.97 (dd, 1H, J ) 2.3, 8.6 Hz),
4.11 (dd, 1H, J ) 2.5, 12.3 Hz), 4.28 (dd, 1H, J ) 5.5, 12.2
Hz), 4.3-4.5 (m, 1H), 4.8 (d, 1H, J ) 1.4 Hz), 5.0-5.35 (m,
5H), 6.14 (bs, 1H), 7.3-7.4 (m, 5H), 7.9-8.1 (m, 2H). Anal.
(C45H51N3O16) C, H, N.
[O-(2,3,4,6-Tetr a-O-acetyl-r-D-m an n opyr an osyl)-D-ser yl]-
D-glu t a m ic a cid 1-m et h yla m id e 5-b en zyl est er (16d ):
1
yield >100% DMF contained; H NMR (CDCl3) δ 1.95, 1.97,
2.10, 2.15 (4s, 12H), 2.0-2.2 (m, 2H), 2.35-2.62 (m, 2H), 2.78
(d, 3H, J ) 5.0 Hz), 3.5-3.8 (m, 2H), 3.9-4.0 (m, 1H), 4.05-
4.15 (m, 1H), 4.15 (d, 1H, J ) 2.5 Hz), 4.25 (dd, 1H, J ) 5.1,
12.3 Hz), 4.3-4.45 (m, 1H), 4.83 (s, 1H), 5.10 (d, 1H, J ) 12.3
Hz), 5.16 (d, 1H, J ) 12.3 Hz), 5.2-5.3 (m, 3H), 6.40 (bs, 1H),
7.3-7.4 (m, 5H).
Gen er a l P r oced u r e for th e P r ep a r a tion of 17a -d :
[N-(2-Tetr a d ecylh exa d eca n oyl)-O-(2,3,4,6-tetr a -O-a cetyl-
r-D-m a n n op yr a n osyl)-D-ser yl]-L-glu ta m ic Acid 1-Meth yl-
a m id e 5-Ben zyl Ester (17b). 16c (345 mg, 0.517 mM) and
2-tetradecylhexadecanoic acid (234 mg, 0.517 mM) were dis-
solved in DMF by heating and then cooled to room tempera-
ture. WSC (119 mg, 0.62 mM) and HOBt (95 mg, 0.62 mM)
were added to the solution. After the mixture stirred for 18.5
h, AcOEt (120 mL) was added to the solution, and the mixture
was washed with 1 N HCl, saturated sodium hydrogen
carbonate, and brine successively, dried (MgSO4), and concen-
trated. The resulting residue was purified by thin-layer
chromatography developing with 25:1 CHCl3/methanol to give
17b (347 mg, 60.9%): 1H NMR (CDCl3) δ 0.85-1.0 (m, 6H),
1.1-1.6 (m, 52H), 1.96, 2.02, 2.11, 2.13 (4s, 12H), 1.9-2.6 (m,
4H), 2.78 (d, 3H, J ) 4.8 Hz), 3.51 (dd, 1H, J ) 5.3, 9.4 Hz),
4.02 (bs, 1H), 4.12-4.39 (m, 3H), 4.4-4.5 (m, 1H), 4.55-4.7
(m, 1H), 4.79 (s, 1H), 5.1 (d, 1H, J ) 12.3 Hz), 5.15 (d, 1H, J
) 12.3 Hz), 5.2-5.3 (m, 3H), 6.37 (q, 1H, J ) 4.9 Hz), 6.48 (d,
1H, J ) 7.0 Hz), 7.1 (d, 1H, J ) 7.8 Hz), 7.35 (s, 5H). Anal.
(C60H99N3O15) C, H, N.
[N-(2-Tetr a d ecylh exa d eca n oyl)-r-D-m a n n op yr a n osyl)-
L-ser yl]-L-glu ta m ic a cid 1-m eth yla m id e (5c): yield 84.0%
1
as a white powder; mp 168 °C; H NMR (DMSO-d6) δ 0.75-
0.9 (m, 6H), 1.0-1.55 (m, 52H), 1.55-2.0 (m, 2H), 2.0-2.3 (m,
3H), 2.56 (d, 3H, J ) 4.5 Hz), 3.2-3.8 (m, 8H), 4.1-4.25 (m,
1H), 4.35-4.5 (m, 1H), 4.61 (s, 1H), 7.67 (q, 1H, J ) 4.5 Hz),
7.80 (d, 1H, J ) 7.9 Hz), 7.98 (d, 1H, J ) 8.0 Hz). Anal.
(C45H85N3O11) C, H, N.
[N-(2-Tetr a d ecylh exa d eca n oyl)-r-D-m a n n op yr a n osyl)-
D-ser yl]-D-glu ta m ic a cid 1-m eth yla m id e (5d ): yield 78.0%
1
as a white powder; H NMR (DMSO-d6) δ 0.85-0.9 (m, 6H),
1.05-1.55 (m, 52H), 1.6-2.0 (m, 2H), 2.1-2.3 (m, 3H), 2.60
(d, 3H, J ) 4.4 Hz), 3.3-3.7 (m, 8H), 4.15-4.3 (m, 2H), 4.5-
4.6 (m, 1H), 4.62 (s, 1H), 7.71 (q, 1H, J ) 4.4 Hz), 7.83 (d, 1H,
J ) 7.5 Hz), 7.95 (d, 1H, J ) 7.9 Hz). Anal. (C45H85N3O11) C,
H, N.
Gen er a l P r oced u r e for th e P r ep a r a tion of Dip ep tid e
Glycosyl Accep tor s 6a -f: N-(ter t-Bu toxyca r bon yl)-D-
ser yl-L-glu ta m ic Acid 1-Meth yla m id e 5-Ben zyl Ester
(6b). To the N-(tert-butoxycarbonyl)-L-glutamic acid 1-me-
thylamide 5-benzyl ester (10.0 g, 28.5 mM) was added 4 N HCl/
1,4-dioxane (30 mL), and the solution was stirred for 0.5 h at
room temperature. The mixture was concentrated, and the
residue was dissolved in DMF (200 mL), added to Et3N (5.4
mL), and then cooled to 0 °C. WSC (6.6 g, 34.2 mM) and HOBt
(5.2 g, 34.2 mM) were added to the mixture, and this was
stirred for 18 h at room temperature. The solvent was
removed under reduced pressure, and the oily residue was
dissolved in AcOEt (300 mL), washed with 0.1 N HCl,
saturated sodium hydrogen carbonate, and brine, dried (Mg-
[N-(2-Tetr adecylh exadecan oyl)-O-(2,3,4,6-tetr a-O-acetyl-
r-D-m a n n op yr a n osyl)-L-ser yl]-D-glu ta m ic a cid 1-m eth yl-
1
a m id e 5-ben zyl ester (17a ): yield 62.0%; H NMR (CDCl3)
δ 0.7-1.0 (m, 6H), 1.0-1.6 (m, 52H), 1.96, 2.01, 2.09, 2.13 (4s,
12H), 2.3-2.65 (m, 2H), 2.77 (d, 3H, J ) 4.8 Hz), 3.7-4.0 (m,
1H), 4.10 (dd, 1H, 2.4, 12.2 Hz), 4.26 (dd, 1H, J ) 5.1, 12.3
Hz), 4.3-4.5 (m, 2H), 4.85 (s, 1H), 5.0-5.35 (m, 5H), 6.38 (d,
1H, J ) 7.1 Hz), 6.49 (d, 1H, J ) 4.6 Hz), 7.20 (d, 1H, J ) 7.9
Hz), 7.25-7.45 (m, 5H). Anal. (C60H99N3O15) C, H, N.
[N-(2-Tetr adecylh exadecan oyl)-O-(2,3,4,6-tetr a-O-acetyl-
r-D-m a n n op yr a n osyl)-L-ser yl]-L-glu ta m ic a cid 1-m eth yl-
a m id e 5-ben zyl ester (17c): yield 55.8% as a crystal; mp

4286 J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22
Tsukida et al.
SO4), and concentrated. The precipitate was filtered, and
recrystallization from AcOEt/n-hexane gave 6b (10.3 g, 82.5%)
as a white crystal: [R]D -5° (c ) 0.1, CHCl3); mp 127-128 °C;
1H NMR (DMSO-d6) δ 1.38 (s, 9H), 1.65-1.85 (m, 1H), 1.90-
2.15 (m, 1H), 2.30-2.40 (m, 2H), 2.58 (d, 3H, J ) 4.6 Hz), 3.54
(t, 2H, J ) 5.6 Hz), 3.85-4.00 (m, 1H), 4.13-4.29 (m, 1H),
4.88 (t, 1H, J ) 5.6 Hz), 5.07 (s, 2H), 6.82 (d, 1H, J ) 6.9 Hz),
7.30-7.45 (m, 5H), 7.77 (d, 1H, J ) 4.6 Hz), 8.03 (d, 1H, J )
8.3 Hz). Anal. (C21H31N3O7) C, H, N.
N-(ter t-Bu toxyca r bon yl)-D-ser yl-L-a sp a r tic a cid 1-m e-
th yla m id e 5-ben zyl ester (6e): yield 94.3%, contained DMF;
1H NMR (CDCl3) δ 1.44 (s, 9H), 2.65-2.80 (m, 1H), 2.75 (d,
3H, J ) 4.8 Hz), 3.15 (dd, 1H, J ) 4.9, 17.2 Hz), 3.70-3.85
(m, 1H), 3.90-4.15 (m, 2H), 4.75-4.90 (m, 1H), 5.10 (d, 1H, J
) 12.2 Hz), 5.15 (d, 1H, J ) 12.2 Hz), 5.63 (bs, 1H), 6.91 (bs,
1H), 7.30-7.45 (m, 5H), 7.52 (d, 1H, J ) 8.0 Hz).
0.8-0.95 (m, 6H), 1.12 (d, 3H, J ) 6.5 Hz), 1.15-1.7 (m, 52H),
1.75-1.92 (m, 1H), 1.92-2.1 (m, 1H), 2.1-2.3 (m, 2H), 2.3-
2.6 (m, 2H), 2.66 (d, 3H, J ) 4.8 Hz), 3.45-3.65 (m, 2H), 3.78-
3.95 (m, 3H), 4.08 (dd, 1H, J ) 4.2,10.2 Hz), 4.15-4.25 (m,
2H), 4.25-4.4 (m, 1H), 4.6-5.0 (m, 7H), 5.05 (d, 1H, J ) 11.9
Hz), 5.1 (d, 1H, J ) 12.3 Hz), 6.0-6.1 (m, 1H), 6.47 (d, 1H, J
) 6.7 Hz), 7.15-7.4 (m, 20H). Anal. (C73H109N3O10) C, H, N.
[N-(2-Un d ecyltr id eca n oyl)-O-(2,3,4-tr i-O-ben zyl-r-L-fu -
cop yr a n osyl)-D-ser yl]-L-glu t a m ic a cid 1-m et h yla m id e
5-ben zyl ester (29): yield 56.2% as a white crystal; mp 127-
128 °C; 1H NMR (CDCl3) δ 0.8-0.95 (m, 6H), 1.12 (d, 3H, J )
6.5 Hz), 1.20-1.75 (m, 40H), 1.9-2.1 (m, 1H), 2.1-2.3 (m, 1H),
2.3-2.6 (m, 2H), 2.66 (d, 3H, J ) 4.8 Hz), 3.45-3.70 (m, 2H),
3.75-3.95 (m, 3H), 4.08 (dd, 1H, J ) 3.5, 9.9 Hz), 4.25-4.50
(m, 2H), 4.55-5.0 (m, 7H), 5.05 (d, 1H, J ) 12.5 Hz), 5.10 (d,
1H, J ) 12.3 Hz), 6.52 (d, 1H, J ) 5.8 Hz), 7.15-7.50 (m, 20H).
Anal. (C67H97N3O10) C, H, N.
[N-P a lm ityl-O-(2,3,4-tr i-O-ben zyl-r-L-fu cop yr a n osyl)-
D-ser yl]-L-glu ta m ic a cid 1-m eth yla m id e 5-ben zyl ester
(30): yield 58.0% as a white crystal; mp 118-120 °C; 1H NMR
(CDCl3) δ 0.8-0.95 (m, 6H), 1.12 (d, 3H, J ) 6.5 Hz), 1.15-
1.40 (m, 24H), 1.45-1.75 (m, 2H), 1.8-1.95 (m, 1H), 2.1-2.3
(m, 3H), 2.3-2.6 (m, 2H), 2.67 (d, 3H, J ) 3.2 Hz), 3.55 (dd,
1H, J ) 8.5, 11.1 Hz), 3.60-3.65 (m, 1H), 3.80-3.95 (m, 3H),
4.08 (dd, 1H, J ) 2.9, 10.3 Hz), 4.25-4.45 (m, 2H), 4.55-4.95
(m, 7H), 5.05 (d, 1H, J ) 12.3 Hz), 5.10 (d, 1H, J ) 12.3 Hz),
6.10 (bs, 1H), 6.52 (d, 1H, J ) 6.4 Hz), 7.15-7.40 (m, 20H).
Anal. (C59H81N3O10) C, H, N.
[N-(2-Tetr a d ecylh exa d eca n oyl)-O-(2,3,4-tr i-O-ben zyl-
r-L-fu cop yr a n osyl)-D-ser yl]-L-a sp a r tic a cid 1-m eth yla -
m id e 5-ben zyl ester (31): yield 75.7% as a white crystal; mp
128-129 °C; 1H NMR (CDCl3) δ 0.8-0.95 (m, 6H), 1.12 (d, 3H,
J ) 6.5 Hz), 1.15-1.60 (m, 52H), 1.85-2.1 (m, 1H), 2.63 (d,
3H, J ) 4.7 Hz), 2.75-3.0 (m, 2H), 3.52 (dd, 1H, J ) 7.7, 11.0
Hz), 3.59 (d, 1H, J ) 1.6 Hz), 3.75-3.95 (m, 3H), 4.06 (dd, 1H,
J ) 3.6, 10.1 Hz), 4.30-4.45 (m, 1H), 4.55-5.0 (m, 7H), 5.10
(s, 2H), 6.45-6.6 (m, 2H), 7.20-7.45 (m, 20H). Anal.
(C72H107N3O10) C, H, N.
N-(ter t-Bu toxycar bon yl)-D-ser yl-L-glu tam ic acid 1-m eth -
yl 5-ben zyl d iester (6f): yield 84.7% as a syrup; 1H NMR
(CDCl3) δ 1.45 (s, 9H), 1.95-2.15 (m, 1H), 2.15-2.35 (m, 1H),
2.40-2.55 (m, 2H), 3.55-3.70 (m, 1H), 3.72 (s, 3H), 4.00-4.25
(m, 2H), 4.55-4.70 (m, 1H), 5.12 (s, 2H), 5.57 (d, 1H, J ) 7.3
Hz), 7.25-7.45 (m, 5H). Anal. (C21H30N2O8) C, H, N.
Gen er a l P r oced u r e for th e P r ep a r a tion of 23-25 by
Glycosyla tion of 6b,e,f w ith 2,3,4-Tr i-O-ben zylfu cose
Don or 22: [N-(ter t-Bu toxyca r bon yl)-O-(2,3,4-tr i-O-ben -
zyl-r-L-fu copyr an osyl)-D-ser yl]-L-glu tam ic Acid 1-Meth yl-
a m id e 5-Ben zyl Ester (23). A mixture of molecular sieves
(4 Å, 2.0 g), AgOTf (0.59 g, 2.28 mM), and SnCl2 (0.43 g, 2.28
mM) in CHCl3 (15 mL) was stirred for 5 h under nitrogen
atmosphere at room temperature and cooled to -40-50 °C.
TMU (0.66 g, 5.70 mM), 2,3,4-tri-O-benzyl-L-fucosyl fluoride
(22) (1.00 g, 2.28 mM) in CHCl3 (1 mL), and compound 6b
(0.50 g, 1.14 mM) in CHCl3 (3 mL) were added to the mixture
successively and stirred for 1 h at the same temperature and
for 20 h while gradually returning to room temperature. The
precipitate was filtered off, and the filtrate was concentrated.
The residue was purified by thin-layer chromatography de-
veloping with 1:1 AcOEt/cyclohexane to give 23 (0.60 g, 62.0%)
as a white crystal: [R]D -59° (c ) 0.1, CHCl3); mp 77-81 °C;
1H NMR (CDCl3) δ 1.13 (d, 3H, J ) 6.5 Hz), 1.43 (s, 9H), 1.7-
2.1 (m, 1H), 2.1-2.3 (m, 1H), 2.3-2.6 (m,2H), 2.67 (d, 3H, J )
4.8 Hz), 3.5-3.7 (m, 2H), 3.8-4.0 (m, 3H), 4.0-4.25 (m, 1H),
4.25-4.45 (m, 1H), 4.5-5.0 (m, 7H), 5.0-5.2 (m, 2H), 5.5 (bs,
[N-(2-Tetr a d ecylh exa d eca n oyl)-O-(2,3,4-tr i-O-ben zyl-
r-L-fu cop yr a n osyl)-D-ser yl]-L-glu t a m ic a cid 1-m et h yl
5-ben zyl d iester (32): yield 75.7% as a syrup; 1H NMR
(CDCl3) δ 0.8-0.95 (m, 6H), 1.10 (d, 3H, J ) 6.5 Hz), 1.2-
1.65 (m, 52H), 1.75-2.1 (m, 2H), 2.1-2.3 (m, 1H), 2.3-2.55
(m, 2H), 3.45 (dd, 1H, J ) 9.3, 11.2 Hz), 3.57 (d, 1H, J ) 1.8
Hz), 3.66 (s, 3H), 3.75-3.9 (m, 2H), 3.86 (dd, 1H, J ) 5.7, 11.5
Hz), 4.06 (dd, 1H,J ) 3.7, 10.1 Hz), 4.4-4.55 (m, 1H), 4.55-
5.0 (m, 7H), 5.04 (d, 1H, J ) 12.5 Hz), 5.09 (d, 1H, J ) 12.5
Hz), 6.50 (d, 1H, J ) 6.3 Hz), 7.20-7.40 (m, 20H). Anal.
(C73H108N2O11) C, H, N.
1H), 6.07 (bs, 1H), 7.1-7.45 (m, 20H). Anal. (C48H59N3O11
C, H, N.
)
[N-(ter t-Bu toxyca r bon yl)-O-(2,3,4-tr i-O-ben zyl-r-L-fu -
cop yr a n osyl)-D-ser yl]-L-a sp a r t ic a cid 1-m et h yla m id e
1
5-ben zyl ester (24): yield 84.0% as a crude syrup; H NMR
(CDCl3) δ 1.13 (d, 3H, J ) 6.4 Hz), 1.43 (s, 9H), 2.63 (d, 3H, J
) 4.8 Hz), 3.55-3.70 (m, 2H), 3.70-4.00 (m, 3H), 4.00-4.20
(m, 2H), 4.55-5.00 (m, 8H), 5.10 (s, 2H), 7.20-7.45 (m, 20H).
[N-(ter t-Bu toxyca r bon yl)-O-(2,3,4-tr i-O-ben zyl-r-L-fu -
cop yr a n osyl)-D-ser yl]-L-glu ta m ic a cid 1-m eth yl 5-ben zyl
d iester (25): crude 25 was used without further purification.
Gen er a l P r oced u r e for th e P r ep a r a tion of 3b a n d 18-
21: [N-(2-Tetr a d ecylh exa d eca n oyl)-O-(r-L-fu cop yr a n o-
syl)-D-ser yl]-L-glu ta m ic Acid 1-Meth yla m id e (3b). To a
solution of 28 (80 mg, 0.067 mM) in ethanol (50 mL) was added
20% Pd(OH)2/C (80 mg), and the mixture was stirred for 4 h
under hydrogen (3-4 atmospheric) pressure. The precipitate
was filtered off, and the filtrate was concentrated in vacuo.
The residue was crystallyzed with water to give 3b (46 mg,
82.5%) as a white crystal: [R]D -62° (c ) 0.1, MeOH); mp 179-
Gen er a l P r oced u r e for th e P r ep a r a tion of 28-32:
[N-(2-Tetr a d ecylh exa d eca n oyl)-O-(2,3,4-tr i-O-ben zyl-r-L-
fu cop yr a n osyl)-D-ser yl]-L-glu ta m ic Acid 1-Meth yla m id e
5-Ben zyl Ester (28). To a solution of 23 (0.5 g, 0.59 mM) in
CH2Cl2 (10 mL) was added TFA (10 mL) at 0 °C, and the
mixture was stirred for 2 h at room temperature. Concen-
trated, the residue was dissolved in AcOEt (100 mL), washed
with saturated sodium carbonate, and dried (MgSO4), and the
solvent was removed in vacuo. The residue was dissolved in
DMF (50 mL), and 2-tetradecylhexadecanoic acid (294 mg, 0.65
mM) was added to the solution. The mixture was dissolved
by heating and then cooled to room temperature. WSC (170
mg, 0.89 mM) and HOBt (136 mg, 0.89 mM) were added to
the solution. After the mixture was stirred for 20 h, AcOEt
(100 mL) was added to the solution, washed with 1 N HCl,
saturated sodium hydrogen carbonate, and brine successively,
dried (MgSO4), and concentrated. The residue was purified
by thin-layer chromatography developing with 25:1 CHCl3/
methanol to give 28 (456 mg, 65.0%) as a white crystal: [R]D
-44° (c ) 0.1, CHCl3); mp 118-120 °C; 1H NMR (CDCl3) δ
1
183 °C; H NMR (DMSO-d6) δ 0.75-0.9 (m, 6H), 1.06 (d, 3H,
J ) 6.4 Hz), 1.1-1.55 (m, 52H), 1.6-1.8 (m, 1H), 1.8-2.1 (m,
1H), 2.1-2.3 (m, 3H), 2.58 (d, 3H, J ) 4.5 Hz), 3.4-3.6 (m,
3H), 3.6-3.85 (m, 2H), 4.1-4.3 (m, 1H), 4.34 (d, 1H, J ) 4.5
Hz), 4.4-4.55 (m, 1H), 7.45 (d, 1H, J ) 4.1 Hz), 7.97 (d, 1H, J
) 7.0 Hz), 8.05 (d, 1H, J ) 8.0 Hz). Anal. (C45H85N3O10) C,
H, N.
[N-(2-Un d ecylt r id eca n oyl)-O-(r-L-fu cop yr a n osyl)-D-
ser yl]-L-glu ta m ic a cid 1-m eth yla m id e (18): yield 84.3% as
1
a white powder; mp 168-170 °C dec; H NMR (DMSO-d6) δ
0.75-0.95 (m, 6H), 1.05 (d, 3H, J ) 6.4 Hz), 1.0-1.55 (m, 40H),
1.6-1.8 (m, 1H), 1.8-2.1 (m, 1H), 2.1-2.3 (m, 3H), 2.57 (d,
3H, J ) 4.5 Hz), 3.6-3.85 (m, 3H), 4.1-4.3 (m, 1H), 4.3-4.55
(m, 2H), 4.40 (d, 1H, J ) 4.5 Hz), 4.64 (s, 1H), 7.79 (q, 1H, J
) 4.6 Hz), 8.04 (d, 1H, J ) 7.3 Hz), 8.09 (d, 1H, J ) 8.4 Hz);
MS spectrum 766 (M + Na)+. Anal. (C39H73N3O10) C, H, N.

Studies on Selectin Blockers
J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 22 4287
(4) Varki, A. Selectin ligands: will the real ones please stand up?
J . Clin. Invest. 1997, 99, 158-162.
[N-P a lm it yl-O-(r-L-fu cop yr a n osyl)-D-ser yl]-L-glu t a m -
ic a cid 1-m eth yla m id e (19): yield 83.0% as a white powder;
mp 155-158 °C dec; 1H NMR (DMSO-d6) δ 0.75-0.95 (m, 3H),
1.05 (d, 3H, J ) 6.5 Hz), 1.1-1.35 (m, 24H), 1.4-1.55 (m, 2H),
1.6-1.8 (m, 1H), 1.85-2.05 (m, 1H), 2.05-2.3 (m, 4H), 2.58
(d, 3H, J ) 4.5 Hz), 3.4-3.7 (m, 5H), 3.75 (q, 1H, J ) 6.6 Hz),
4.1-4.25 (m, 1H), 4.25-4.6 (m, 4H), 4.64 (s, 1H), 7.78 (q, 1H,
J ) 4.6 Hz), 7.98 (d, 1H, J ) 7.1 Hz), 8.14 (d, 1H, J ) 8.2 Hz),
12.05 (bs, 1H);MS spectrum 654 (M +Na)+. Anal. (C31H57N3O10)
C, H, N.
[N-(2-Tetr a d ecylh exa d eca n oyl)-O-(r-L-fu cop yr a n osyl)-
D-ser yl]-L-a sp a r tic a cid 1-m eth yla m id e (20): yield 86.7%
as a pale-yellow powder; mp 60-63 °C dec; 1H NMR (DMSO-
d6) δ 0.75-0.95 (m, 6H), 1.05 (d, 3H, J ) 6.4 Hz), 1.1-1.5 (m,
52H), 2.1-2.3 (m, 2H), 3.3-3.8 (m, 5H), 4.3-4.5 (m, 2H), 4.5-
4.6 (m, 1H); MS spectrum 836 (M + Na)+. Anal. (C44H83N3O10)
C, H, N.
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Studies on Selectin Blocker 1: Structure-Activity Relationships
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K.; Ohashi, M.; Kondo, H.; Ishida, H.; Kiso, M.; Hasegawa, A.
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Selectin. J . Med. Chem. 1996, 39, 4547-4553. (d) Yoshino, K.;
Ohmoto, H.; Kondo, N.; Tsujishita, H.; Hiramatsu, Y.; Inoue,
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Selectin Blockers 4: Structure-Function Relationships of Sul-
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and L-Selectin Binding. J . Med. Chem. 1997, 40, 455-462. (e)
Studies on Selectin Blockers 5: Design, Synthesis, and Biological
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[N-(2-Tetr a d ecylh exa d eca n oyl)-O-(r-L-fu cop yr a n osyl)-
D-ser yl]-L-glu ta m ic a cid 1-m eth yl ester (21): yield 93.7%
as a pale-yellow powder; mp 40-41 °C dec; 1H NMR (DMSO-
d6) δ 0.75-0.95 (m, 6H), 1.05 (d, 3H, J ) 6.4 Hz), 1.1-1.55
(m, 52H), 1.65-1.85 (m, 1H), 1.85-2.1 (m, 1H), 2.1-2.3 (m,
3H), 3.35-3.65 (m, 5H), 3.62 (s, 3H), 3.73 (q, 1H, J ) 6.6 Hz),
4.2-4.4 (m, 1H), 4.39 (d, 1H, J ) 4.4 Hz), 4.5-4.65 (m, 1H),
4.65 (s, 1H), 7.89 (d, 1H, J ) 8.3 Hz), 8.27 (d, 1H, J ) 7.7 Hz),
12.12 (s, 1H);MS spectrum 851 (M +Na)+. Anal. (C45H84N2O11
C, H, N.
)
[N-(2-Tetr a d ecylh exa d eca n oyl)-O-(r-L-fu cop yr a n osyl)-
D-ser yl]-L-glu ta m ic Acid 1-Meth yla m id e 5-Meth yl Ester
(33). To a solution of 3b (100 mg, 0.12 mM) in methanol-
toluene (1:1, 10 mL) was added 2 M TMSCH2N2/n-hexane (0.18
mL, 0.36 mM), and the mixture was stirred for 30 min at room
temperature. Concentrated under reduced pressure, the
resulting precipitate was collected with Et2O to give 33 (96
mg, 95.0%) as a pale-yellow powder: mp 100-102 °C; 1H NMR
(DMSO-d6) δ 0.75-0.95 (m, 6H), 1.06 (d, 3H, J ) 6.4 Hz), 1.1-
1.6 (m, 52H), 1.65-1.85 (m, 1H), 1.85-2.1 (m, 1H), 2.15-2.4
(m, 4H), 2.57 (d, 3H, J ) 4.4 Hz), 3.4-3.8 (m, 5H), 3.57 (s,
3H), 4.1-4.3 (m, 1H), 4.4-4.55 (m, 1H), 4.65 (s, 1H), 7.81 (q,
1H, J ) 4.4 Hz), 8.02 (d, 1H, J ) 7.2 Hz), 8.08 (d, 1H, J ) 8.2
Hz); MS spectrum 864 (M + Na)+. Anal. (C46H87N3O10) C, H,
N.
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