Novel synthetic inhibitors of selectin-mediated cell adhesion: Synthesis of 1,6-bis[3-(3-carboxymethylphenyl)-4-(2-α-D-mannopyranosyloxy)phenyl] hexane (TBC1269)

Article abstract of DOI:10.1021/jm9704917

Reports of a high-affinity ligand for E-selectin, sialyl di-Lewis(x) (sLe(x)Le(x), 1), motivated us to incorporate modifications to previously reported biphenyl-based inhibitors that would provide additional interactions with the protein. These compounds were assayed for the ability to inhibit the binding of sialyl Lewis(x) (sLe(x), 2) bearing HL-60 cells to E-, P-, and L- selectin fusion proteins. We report that dimeric or trimeric compounds containing multiple components of simple nonoligosaccharide selectin antagonists inhibit sLe(x)-dependent binding with significantly enhanced potency over the monomeric compound. The enhanced potency is consistent with additional binding interactions within a single selectin lectin domain; however, multivalent interaction with multiple lectin domains as a possible alternative cannot be ruled out. Compound 15e (TBC1269) showed optimal in vitro activity from this class of antagonists and is currently under development for use in the treatment of asthma.

Full text of DOI:10.1021/jm9704917

1099
J . Med. Chem. 1998, 41, 1099-1111
Novel Syn th etic In h ibitor s of Selectin -Med ia ted Cell Ad h esion : Syn th esis of
1,6-Bis[3-(3-ca r boxym eth ylp h en yl)-4-(2-r-D-m a n n op yr a n osyloxy)p h en yl]h exa n e
(TBC1269)
Timothy P. Kogan,* Brian Dupre´, Huong Bui, Kathy L. McAbee, J amal M. Kassir, Ian L. Scott, Xin Hu,†
Peter Vanderslice,† Pamela J . Beck,†,‡ and Richard A. F. Dixon
Departments of Chemistry & Biophysics and Immunology, Texas Biotechnology Corporation, 7000 Fannin, Suite 1920,
Houston, Texas 77030
Received J uly 29, 1997
Reports of a high-affinity ligand for E-selectin, sialyl di-Lewisx (sLexLex, 1), motivated us to
incorporate modifications to previously reported biphenyl-based inhibitors that would provide
additional interactions with the protein. These compounds were assayed for the ability to inhibit
the binding of sialyl Lewisx (sLex, 2) bearing HL-60 cells to E-, P-, and L-selectin fusion proteins.
We report that dimeric or trimeric compounds containing multiple components of simple
nonoligosaccharide selectin antagonists inhibit sLex-dependent binding with significantly
enhanced potency over the monomeric compound. The enhanced potency is consistent with
additional binding interactions within a single selectin lectin domain; however, multivalent
interaction with multiple lectin domains as a possible alternative cannot be ruled out.
Compound 15e (TBC1269) showed optimal in vitro activity from this class of antagonists and
is currently under development for use in the treatment of asthma.
In tr od u ction
a number of antagonists have been reported, which are
mainly represented by oligosaccharides6 and glycosy-
lated peptides.7 Both classes of compounds may suffer
from a number of problems regarding development as
pharmaceutical agents, particularly regarding cost of
goods, stability, and pharmacokinetic half-life. Our
efforts in this area resulted in the identification of a
novel class of low molecular weight, non-oligosaccharide
biphenyl-based compounds exemplified by 3 (Figure 1),
which possess significant advantages over oligosaccha-
ride-based inhibitors.8 Benefits include fewer chiral
centers, providing easier synthetic access and the
potential for lower cost of materials, as well as increased
thermal and chemical stability, and the potential for
oral bioavailability. These compounds were designed
to utilize mannose and carboxylic acid functionalities
as replacements for the critical fucose and sialic acid
groups of sLex, respectively, and a biphenyl unit to
replace the lactosamine core. No direct evidence exists
regarding the specific set of interactions made between
E-selectin and sLex; however several pieces of circum-
stantial evidence9 support our earlier hypothesis in
which the fucose is coordinated to calcium, and the sialic
acid is oriented toward Arg-97 and Lys-99.10,11 Regard-
less of the actual mode of binding used by sLex, 3 can
be considered a good functional mimetic, since it is a
competitive inhibitor of sLex binding to E-, P-, and
L-selectin.8 By comparison, compounds lacking either
the acid group or the mannose group in this series were
found to be essentially devoid of activity against each
of the three selectins.
The selectins participate in the adhesion of leukocytes
to endothelial cells by mediating the initial rolling of
leukocytes on the lumen wall of blood vessels. They are
a family of three structurally related calcium-dependent
cell adhesion molecules which are expressed on the
surface of activated vascular endothelial cells (E- and
P-selectin), activated platelets (P-selectin), and leuko-
cytes (L-selectin).1 These proteins function by binding
sialyl Lewisx (sLex, 2, Figure 1)2,3 and related oligosac-
charides which are present on the surface of neutrophils,
monocytes, a subset of T lymphocytes, eosinophils,
basophils, and endothelial cells. One of the initial
events following injury is the recruitment of leukocytes
to activated endothelium in the vicinity of the injury
and is mediated by the interaction between sLex on the
leukocyte and E- and P-selectin expressed on activated
endothelial cells. Amplification of adherance is achieved
by L-selectin-mediated interactions between circulating
leukocytes and adhered leukocytes. However, the ac-
cumulation of excess leukocytes, or inappropriate leu-
kocyte recruitment, may contribute to the development
of several disease states. Thus, the ability to regulate
the interactions between the selectins and sLex or other
ligands could have potential utility in a number of
clinical conditions such as asthma, reperfusion injury,
psoriasis, cancer, lupus, ARDS, septic shock, rheumatoid
arthritis, and other cell adhesion-mediated diseases.4
The key functional groups of sLex necessary for
interaction with the selectins have been identified,5 and
As part of our ongoing research in this area, we
sought to investigate rational approaches to the genera-
tion of selectin inhibitors with increased potency. Re-
cently there have been reports of more complex oligosac-
charides that may represent relevant in vivo E-selectin
* To whom correspondence should be addressed; e-mail:
tkogan@tbc.com. Tel: (713) 796-8822. Fax: (713) 796-8232.
† Department of Immunology.
‡ Current address: Institute of Molecular Medicine for the Preven-
tion of Human Disease, The University of Texas, Houston Health
Science Center, 2121 W. Holcombe, Houston, TX 77030.
S0022-2623(97)00491-3 CCC: $15.00 © 1998 American Chemical Society
Published on Web 02/28/1998

1100 J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 7
Kogan et al.
F igu r e 1. Characterized E-selectin ligands sLexLex1 and sLex2, and the biphenyl-based glycomimetic 3.
Sch em e 1a
a
Reagents: (i) 3-bromophenylacetyl chloride, AlCl3; (ii) 2-methoxyphenylboronic acid, Pd(0), aqueous Na2CO3; (iii) (a) aqueous KOH;
(b) N2H4; (c) BBr3; (d) MeOH, H+; (iv) R-D-mannose pentaacetate, BF3‚Et2O; (v) aqueous LiOH.
ligands of high potency, as well as reports that multi-
valent sLex presentation may be important in obtaining
high-affinity interactions.12 We report here the results
of investigations into the design of mimetics of higher
affinity selectin ligands which possess increased struc-
tural complexity, as well as preliminary investigations
of potential multivalent analogues of compound 3, which
have led to the development of a class of inhibitors with
increased potency.
explain this discrepancy, including the conformational
biasing of the mobile sialic acid residue in the proximity
of other complex glycoproteins on the cell surface.14
Clearly, an alternative explanation is the presence of
other ligands of higher affinity which remain to be
identified. Along these lines, Parekh has reported the
identification of several tetra-antennary oligosaccharide
structures isolated from U937 cells and human neutro-
phils.12 Upon characterization, these oligosaccharides
were each shown to contain one branch bearing the
extended sialyl di-Lewisx (sLexLex, 1, Figure 1) motif.
These oligosaccharides were reported to bind E-selectin
with much greater affinity than either sLex or sLea (KI’s
of approximately 1 µM) yet are expressed on less than
3% of cell surface glycoproteins.
We considered that incorporation of an additional
mannose unit via an appropriate spacer may result in
derivatives of 3 that would be mimetics of the more
potent sLexLex oligosaccharide. Preliminary modeling
studies suggested that a biphenylethyl unit would
function suitably, and that the biphenyl ether 4 (Scheme
1) would permit synthetic access, since it would undergo
Friedel-Crafts chemistry selectively at the desired
4-position (para to the methoxy substituent). Thus 4
was treated with 3-bromophenylacetyl chloride and
Design a n d Syn th esis
While some knowledge has recently been gained
regarding the nature of selectin ligands, a clear under-
standing of the structure of all relevant in vivo ligands
for each of the selectins remains elusive.3b,13 For
example, the glycoprotein PSGL-1 has been reported as
a native ligand of both L- and P-selectin and bears the
sLex carbohydrate motif. Further known ligands in-
clude ESL-1 for E-selectin and GlyCAM-1, MAdCAM-
1, and CD34 for L-selectin. The sLex motif was iden-
tified as a natural oligosaccharide capable of selectin
recognition some time ago. However, doubts have
remained about the relevance of a ligand with millimo-
lar potency in in vitro assays to the in vivo adhesion of
leukocytes. Several hypotheses have been offered to

Inhibitors of Selectin-Mediated Cell Adhesion
J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 7 1101
Sch em e 2a
a
Reagents: (i) acid chloride, AlCl3; (ii) BBr3; (iii) R-D-mannose pentaacetate, BF3‚Et2O; (iv) aqueous NaOH or aqueous LiOH; (v) (a)
aqueous LiOH, then N2H4; or Et3SiH, TFA, BF3‚Et2O; or Pd(OH)2/H2; then (b) BBr3; (c) MeOH, H+.
aluminum chloride to afford ketone 6. A Suzuki cou-
pling with 2-methoxyphenylboronic acid,15 followed by
Wolff-Kishner reduction of the ketone, demethylation
of the two methyl ethers and re-esterification of the acid
gave the bis-phenol 8. Glycosylation using R-D-mannose
pentaacetate, followed by hydrolysis of the esters, gave
9. For comparison, a control compound with alternate
spacing of the two mannose groups and carboxylic acid,
the bis[(mannosyloxy)phenyl]valeric acid 10 was pre-
pared by treatment of 4,4′-bis(4-hydroxyphenyl)valeric
acid with R-D-mannose pentaacetate, followed by depro-
tection.
Having established that Friedel-Crafts acylation
provided a facile route to functionalization of the biphen-
yl unit, we recognized that the use of a multivalent acid
chloride would allow an alternative and concise route
to compounds bearing multiple mannose units through
a dimerization or trimerization process. Thus, Friedel-
Crafts acylation of 4 or 5 using a variety of acid chlorides
was used as a convenient route to access these struc-
tures (Scheme 2). Ether cleavage, followed by glycosy-
lation and hydrolysis, produced 13a . Phenols 14a -h
were obtained from 11a -h by one of three different
methods: (a) Wolff-Kishner reduction of the ketone
followed by ether cleavage and re-esterification, (b)
reduction of the ketone using triethylsilane in the
presence of boron trifluoride etherate and trifluoroacetic
acid followed by ether cleavage, or (c) catalytic reduction
followed by ether cleavage with boron tribromide. Use
of the ethyl rather than methyl ester (starting from 5
rather than 4) suppressed ester cleavage which was
occasionally observed during demethylation of the phe-
nol. Finally, compounds 15a -h were obtained by
standard glycosylation, followed by hydrolysis. During
the preparation of 15e, a sample of the monoglycoside
16 was also isolated and characterized.
An alternative method was used to prepare derivative
19 (Scheme 3). Commercially available 4-(4-methoxy-
phenyl)butyric acid was converted to the acid chloride
with thionyl chloride, and subjected to Friedel-Crafts
acylation with anisole followed by reduction of the
ketone to the symmetrical bis-ether 17. Bis-ortho-
lithiation, boronation, and hydrolysis gave the inter-
mediate bis-boronic acid, which was subjected to Suzuki
coupling with methyl 3-bromophenylacetate to give bis-
biphenyl 18. Demethylation using boron tribromide,
followed by glycosylation and then hydrolysis, gave 19.
The sulfide and sulfone spaced compounds 22 and 23,
respectively, were obtained from the propanoyl bromide
11i (Scheme 4). Reduction of the ketone followed by

1102 J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 7
Kogan et al.
Sch em e 3a
a
Reagents: (i) SOCl2; (ii) anisole, AlCl3; (iii) Et3SiH, TFA, BF3‚Et2O; (iv) n-BuLi, THF, TMEDA; (v) (a) B(OMe)3; (b) H3O+; (vi) methyl
3-bromophenyl acetate, Pd(0), K3PO4; (vii) BBr3; (viii) R-D-mannose pentaacetate, BF3‚Et2O; (ix) aqueous LiOH.
Sch em e 4a
a
Reagents: (I) Et3SiH, TFA, BF3‚Et2O; (ii) (a) NaH, 1,3-propanedithiol, (b) BBr3, (c) R-D-mannose pentaacetate, BF3‚Et2O; (iii) aqueous
NaOH or aqueous LiOH; (iv) m-CPBA.
Sch em e 5a
end, a series of derivatives were prepared as outlined
in Scheme 7. For the amine series of compounds, the
more stable pivalate protecting group was used for the
mannose hydroxyls to prevent acyl migration onto the
basic nitrogen.16 The pivotal intermediate for these, 35,
was prepared by Friedel-Crafts acylation of 34 with
succinic anhydride. Reduction of the ketone, generation
of the acid chloride, and condensation with a number
of amines gave 36a -e. Hydrolysis of the esters then
gave the target amides 37a -e. Alternatively, amide
36a was reduced with borane in THF, and then hydro-
lyzed to provide the target amine 38a . Compound 39
was obtained by reduction of the ketone and acid
functional groups of 35 with borane in THF, followed
by reaction of the resulting alcohol with cyanuric
chloride and then hydrolysis of the protecting groups
with aqueous base.
a
Reagents: (i) Et3SiH, TFA, BF3‚Et2O; (ii) BBr3; (iii) R-D-
mannose pentaacetate, BF3‚Et2O; (iv) aqueous NaOH.
The methods used to prepare the previous biphenyl
derivatives were unsuitable for access to 42. Due to the
additional oxygen substitution on the biphenyl system,
a single position was no longer preferred in the Friedel-
Crafts acylation; therefore an alternate route was
devised as outlined in Scheme 8. Thus, 1,6-bis(4-
methoxyphenyl)hexane 40 was prepared by Friedel-
Crafts acylation of anisole with adipoyl chloride, fol-
lowed by Wolff-Kishner reduction of the ketones.
Bromination ortho to the methoxy groups with bromine/
ferric chloride followed by cleavage of the methyl ethers
with boron tribromide and glycosylation using R-D-
mannose pentaacetate gave 41. Suzuki coupling of the
bromide 41 with 3-hydroxybenzene boronic acid followed
by alkylation of the phenol with ethyl bromoacetate and
then hydrolysis gave the target compound 42.
In Vitr o Assa y Da ta . The compounds described
above were evaluated for their ability to inhibit the
binding of sLex expressing HL-60 cells to recombinant
fusion proteins bearing the lectin domain of human E-,
P-, or L-selectin, the EGF-like domain, and two comple-
ment repeats fused to the hinge, CH2, and CH3 domains
of mouse IgG, and the results are reported in Table 1.17
Compound 9, which was designed to mimic the sLexLex
motif, displayed improved affinity for E- and L-selectin,
compared with 3. As anticipated, the relative spatial
arrangement of the two mannose units and carboxylic
reaction of the bromide 20 with propane dithiol and then
demethylation and glycosylation gave 21. Hydrolysis
of the esters gave the sulfide 22, while prior oxidation
of the bis-sulfide followed by hydrolysis gave the bis-
sulfone 23.
The bis-glycoside with only a single carboxylate 26
(Scheme 5) was prepared from 11j by reduction of the
ketone groups with triethylsilane, followed by demethyl-
ation which gave 25. Glycosylation of the phenols and
then deprotection gave 26.
An alternative approach permitted the use of the
glycosylated ester 27 (prepared from 5 by ether cleavage
and glycosylation) as an advanced intermediate (Scheme
6). Friedel-Crafts acylation with either trans-3-hex-
enedioic acid chloride, adipoyl chloride, or bromoacetyl
bromide gave the ketones 28-30, respectively. Reduc-
tion of the ketone 28 with triethylsilane, followed by
hydrolysis of the protecting groups, gave 31, while 29
was hydrolyzed directly to provide the diketone 32. A
trimer was prepared by reaction of 30 with 1,3,5-tris-
(mercaptomethyl)benzene and sodium hydride, followed
by hydrolysis of the protecting groups which gave 33.
We sought to further investigate a variety of linking
groups which included amides and amines due to their
potential to address variations of both distance and
relative conformation of the terminal units. Toward this

Inhibitors of Selectin-Mediated Cell Adhesion
J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 7 1103
Sch em e 6a
a
Reagents: (i) acid chloride, AlCl3; (ii) Et3SiH, TFA, BF3‚Et2O; (iii) aqueous NaOH; (iv) NaH, 1,3,5-tris(mercaptomethyl)benzene.
acid were shown to be important, as the control com-
pound 10 was essentially inactive at the concentration
tested.
Discu ssion
While the binding of carbohydrates such as sLex and
related oligosaccharides to proteins is typically weak in
nature, more complex oligosaccharide structures such
as sialyl di-Lewisx show higher affinity. The results
presented here demonstrate that the design of non-
oligosaccharide mimetics of complex oligosaccharides
can result in high-affinity antagonists. From the data
reported it is of interest to consider whether these
compounds are likely to work as functional mimetics of
sialyl di-Lewisx, or if other mechanisms/modes of action
are responsible for their enhanced potency. Although
a definitive answer to this question is not currently
possible, evaluation of the available SAR data provides
for speculation.
The binding data obtained for the series of dimer and
trimer compounds provided additional insight into
structure/activity relationships. Although diketone 13a
could not be tested due to insolubility under the assay
conditions, the diketone 32 displayed improved activity
compared to the parent compound 3. In the series of
compounds 15a ,b,d -h , 19, and 31 the optimum spacing
and orientation appears to be present in 15d for E- and
L-selectin (a pentane linker), and 15e for P-selectin (a
hexane linker). Of note in this series of compounds was
the complete lack of activity of 31 at 3 mM compared
with 15e. These compounds differ only in that 31 has
an (E)-3-hexenyl linkage rather than the hexyl linker
in 15e.
One alternative hypothesis due consideration is that
of multivalent interaction between these compounds and
two independent selectin molecules. Indeed, several
groups have shown significantly enhanced potency
achieved through multiple presentation of selectin
recognizing oligosaccharides on either protein,18 lipo-
somal,19 or polymeric templates.20 The published data
suggests that more than bivalent presentation of ligands
is required to observe significant enhancements in
potency,18 although dimers of sLex have been reported
with 4-6-fold greater activity than the monomer,
perhaps functioning as mimetics of sLexLex.21
The monoglycoside 16 was inactive against E- and
L-selectin at the concentration tested; however, activity
in the P-selectin assay was comparable to that of 3, yet
significantly less potent than 15e. Conversely, the bis-
glycoside monocarboxylate 26 was about as active as
15e in the E- and L-selectin assays and had diminished
activity against P-selectin.
In the series of sulfur-containing compounds 22, 23,
and 33, only 22 showed activity at the concentrations
tested and was P-selectin selective. All of the nitrogen-
containing compounds, 37a -e, 38a , and 39, were inac-
tive at the concentrations tested, except for 37b which
showed a slight improvement for E- and P-selectin
compared to the parent monomeric compound. The
oxygen-containing derivative 15c was selective for P-
selectin, as was the compound with ether linkages
between the biphenyl and carboxylic acid moieties, 42.
None of the trimeric compounds prepared, 15h , 33, 37d ,
or 39, showed improved properties over the preferred
dimeric compounds such as 15e.
The most compelling data to support the action of
these compounds as sialyl di-Lewisx mimetics comes
from the set of compounds that contain varying numbers
of either carboxylates or mannose units present. These
include 9 (two mannose, one carboxylate), 26 (two
mannose, one carboxylate), 16 (one mannose, two car-
boxylates), and 15e (two mannose, two carboxylates).
It was observed that a second carboxylate was not
required for enhanced acivity against each of the selec-

1104 J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 7
Kogan et al.
Sch em e 7a
a
Note: Piv is used as an abbreviation for trimethylacetyl. Reagents: (i) succinic anhydride, AlCl3; (ii) Et3SiH, TFA, BF3‚Et2O; (iii)
SOCl2; (iv) amines a-e, NEt3; (v) BH3‚THF; (vi) NaOMe, then aqueous NaOH; (vii) NaH, cyanuric chloride.
Sch em e 8a
analogue, 31, which contained a six carbon spacer with
a trans olefin, was inactive. These data suggest that
conformations of the alkyl spacer that are required for
enhanced activity are not available to the more rigid
compound 31. Specifically, those conformations in
which the hexyl linker is folded back are required to
enable both mannose units to interact with the same
protein surface. Other variations in length or atom type
in the linker did not lead to improved activity. In fact,
many of those with longer, nitrogen-containing linkers
(37a -e and 38a ) had reduced or no observable activity.
In the series of trimeric compounds investigated (15h ,
33, 37d , or 39), only 15h proved to be effective, while
the others in the series were inactive at the concentra-
tions tested.
a
Reagents: (i) adipoyl chloride, AlCl3; (ii) N2H4, KO-t-Bu,
DMSO; (iii) Br2, FeCl3; (iv) BBr3; (v) R-D-mannose pentaacetate,
BF3‚Et2O; (vi) 3-hydroxyphenyl boronic acid, Pd(0), K3PO4; (vii)
ethyl bromoacetate, Cs2CO3; (viii) aqueous NaOH.
The results presented in this paper, particularly with
regard to the potency of divalent structures, suggest
that both units of compounds such as 15e likely bind
to the same lectin domain. This hypothesis is consistent
with a number of results published from groups working
with other systems in which enhanced potency was
obtained through establishing an additional set of
interactions.22 Preliminary modeling suggests that
compounds of this type may adopt conformations in
which the mannose units occupy positions similar to
tins (compounds 9 and 26), whereas the monomanno-
sylated compound, 16, had significantly reduced activ-
ity.
Additional implications with regard to the mode of
lectin binding adopted by the dimeric compounds are
derived from evaluating the nature of the linking group.
Dimers linked by five, six, or seven saturated carbon
atoms (15d ,e,f) displayed significant improvements in
inhibiting all three selectin proteins compared to the
monomeric compound 3. Surprisingly the more rigid

Inhibitors of Selectin-Mediated Cell Adhesion
J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 7 1105
Ta ble 1. In Vitro Inhibition of sLeX Bearing HL-60 Cells to
Selectin-IgG Fusion Proteins24
IC50(mM) or % inhib at mM
compd
E-selectin
P-selectin
L-selectin
sLeX
3
3.3
3.1
3.4
2.6
3.5
4.1
9
10
1.0
12 at 3
a
2.0
0 at 1
2.5
0.4
0.5
0.5
40 at 1
45 at 0.65
0 at 3
5.0
29 at 2
0 at 3
0.3
2.0
0 at 3
a
2.0
0.225
0.7
0.3
0.07
0.4
40 at 1
0.2
2.0
0.75
20 at 3
a
3.0
3.0
0 at 2
0.3
0.56
0.75
1.0
13a
15a
15b
15c
15d
15e
15f
15g
15h
16
0.5
10 at 3
12 at 2
5.0
0 at 3
0.6
19
22
23
26
2.5
0.5
0 at 3
1.0
31
32
0 at 3
1.0
0 at 3
1.0
0 at 3
3.0
33
4.0
0 at 3
1.5
40 at 3
15 at 3
0 at 3
0 at 3
0 at 3
3.2
0 at 3
0 at 3
1.0
0 at 3
0 at 3
0 at 3
0 at 3
0 at 3
0.54
2.0
37a
37b
37c
37d
37e
38a
39
0 at 3
0 at 3
0 at 3
0 at 3
0 at 3
0 at 3
25 at 3
1.4
F igu r e 2. Views of a low-energy conformation of 1 (top) and
15e (bottom) bound to the E-selectin calcium site. The com-
pounds are color coded to indicate the similar relative position-
ing possible for the carboxylic acids (magenta) and the
mannose and fucose residues (yellow). The E-selectin calcium
is shown as a blue sphere, while the residues Lys-99, Arg-97,
Lys-111, and Arg-108 (blue) are positioned sequentially from
left to right. The distances from the carboxylate carbon to a
centoid between the calcium coordinating hydroxyl groups are
12.07 Å for 1 and 11.24 Å for 15e.
42
a
Compound not soluble at concentration tested
each of the fucose units of sialyl di-Lewisx, as indicated
by a comparison of selected low energy confomations
(Figure 2). A more extensive molecular modeling ap-
proach has been initiated to investigate which modes
of binding of 15e are likely to be preferred, and how
such interactions differ for the three selectin proteins.
Initial studies reveal that 15e can adopt a low-energy
conformation wherein the two mannose groups can be
overlaid with the two fucose residues of 1, while one of
the carboxylates of 15e is positioned in similar proximity
to the sialyl carboxylate (Figure 2). More detailed
studies correlating the conformational flexibility and
relative energies of possible bound conformations are
ongoing and will be published elsewhere.
While the data presented support the notion that the
second carboxylate of 15e is functionally redundant, this
is more than compensated for by simplification of the
synthetic chemistry which results in a symmetrical
molecule. Due to the interesting in vitro data gathered
on 15e, this compound has been evaluated in animal
models of inflammation, and the results from these
studies will be reported elsewhere.23 Current efforts are
directed toward further enhancements of activity in the
monovalent series of compounds, as well as approaches
to a series of compounds with considerably extended in
vivo half-life. Results from these studies, as well as the
clinical development of 15e, will be reported in due
course.
Rainin Dynamax 300 Å 5 µm (21 mm i.d. × 25 cm) C-18
column using a gradient of water and acetonitrile (with 0.1%
TFA) for elution, with UV monitoring at 254 nm. Pure
fractions were lyophilized and analyzed for purity. Proton
NMR spectra were recorded on a J EOL 400 MHz spectrometer
using an internal reference. Infrared spectra were recorded
on a Bruker IFS-25 instrument as thin films on sodium
chloride plates, or as KBr pellets. Melting points were
determined using a Fisher-J ohns hot stage apparatus and are
uncorrected.
3-(2-r-D-Ma n n op yr a n osyloxy-5-(2-(3-(2-r-D-m a n n op y-
r an osyloxyph en yl)ph en yl)eth yl)ph en yl)ph en ylacetic Acid
(9). Step 1. Methyl 3-(2-methoxyphenyl)phenylacetate (4,
0.82 g, 3.2 mmol) and 3-bromophenylacetyl chloride (0.77 g,
3.3 mmol) were dissolved in 1,2-dichloroethane (11 mL) and
chilled in an ice bath. Aluminum chloride (0.88 g, 6.6 mmol)
was added in one portion and the mixture was warmed at 50
°C for 15 min then mixed with ice water. The organic
materials were separated, dried (MgSO4), then concentrated
under reduced pressure which gave 1.68 g of methyl 3-(2-
methoxy-5-(1-oxo-2-(3-bromophenyl)ethyl)phenyl)phenyl-
acetate (6) as a yellow oil which was used without further
purification. 1H NMR (400 MHz, CDCl3): 7.95-8.20 (m, 2H),
7.35-7.45 (m, 6H), 7.29 (br d, J ) 6.9 Hz, 1H), 7.18-7.22 (m,
1H), 7.01 (d, J ) 8.4 Hz, 1H), 4.22 (s, 2H), 3.87 (s, 3H), 3.70
(s, 3H), 3.69 (s, 2H) ppm. IR (NaCl): 1730, 1686, 1262 cm-1
.
Step 2. The crude halide 6 was mixed with (2-methoxyphe-
nyl)boronic acid (0.54 g, 3.55 mmol) in toluene (20 mL), then
tetrakis(triphenylphosphine)palladium(0) (120 mg, 3 mol %)
and 2 N sodium carbonate (6 mL) were added, and the mixture
was degassed. The mixture was heated at reflux for 14 h, then
mixed with water, and extracted with ethyl acetate. The
organic materials were combined, washed with saturated
sodium chloride, dried (MgSO4), and then concentrated under
reduced pressure. Purification by flash chromatography (SiO2,
gradient elution, hexane to 3:1 hexane/ethyl acetate) gave 1.57
g (77%) of methyl 3-(2-methoxy-5-(1-oxo-2-(3-(2-methoxyphe-
Exp er im en ta l Section
Reagents and solvents were used as received unless other-
wise specified. All reactions involving organometallic com-
pounds or other air or moisture sensitive materials were
conducted under a nitrogen atmosphere, in oven-dried glass-
ware. Preparative reverse-phase HPLC was conducted on a

1106 J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 7
Kogan et al.
nyl)phenyl)ethyl)phenyl)phenylacetate (7) as a clear oil. 1H
NMR (400 MHz, CDCl3): 8.00-8.05 (m, 2H), 7.23-7.46 (m,
10H), 6.93-7.03 (m, 3H), 4.29 (s, 2H), 3.86 (s, 3H), 3.75 (s,
3H), 3.69 (s, 3H), 3.65 (s, 2H) ppm. 13C NMR (100 MHz,
CDCl3): 196.3, 172.1, 160.3, 156.3, 138.7, 136.8, 134.2, 133.9,
132.0, 131.0, 130.7, 130.5, 130.4 (3), 129.7, 128.7, 128.4 (4),
128.1, 128.0, 120.9, 111.2, 110.7, 55.9, 55.6, 52.2, 45.6, 41.3
3.51 (m, 6H), 2.28 (m, 2H), 1.94 (m, 2H), 1.52 (s, 3H) ppm. IR
(KBr): 3405, 1710 cm-1. Anal. (C29H38O14‚0.7TFA) C, H.
1,4-Bis[3-(3-ca r boxym eth ylp h en yl)-4-r-D-m a n n op yr a -
n osyloxy]ben zoylben zen e (13a ). Step 1. Ethyl 3-(2-meth-
oxyphenyl)phenylacetate (5, 1.7 g, 6.33 mmol) in 1,2-dichlo-
roethane (5 mL) was added to a suspension of terephthaloyl
chloride (0.61 g, 3.0 mmol) and aluminum chloride (2.54 g, 19.1
mmol) in 1,2-dichloroethane (15 mL). The mixture was stirred
at room temperature under nitrogen for 30 min, then mixed
with ice water, and extracted with dichloromethane. The
organic materials were combined, washed with saturated
sodium chloride, dried (MgSO4), and then concentrated under
reduced pressure. Purification by flash chromatography (SiO2,
gradient elution, 3:1 hexane/ethyl acetate to ethyl acetate) gave
1.0 g (50%) of 11a as a clear oil. 1H NMR (400 MHz, CDCl3):
7.86 (m, 10 H), 7.43 (m, 2H), 7.37 (t, J ) 8.0 Hz, 2H), 7.28 (m,
2H), 7.05 (d, J ) 8.0 Hz, 2H), 4.14 (q, J ) 7.0 Hz, 4H), 3.90 (s,
6H), 3.65 (s, 4H), 1.24 (t, J ) 7.0 Hz, 6H) ppm. IR (NaCl):
ppm. IR (NaCl): 1739, 1674, 1269, 1244 cm-1
.
Step 3. The ketone (7, 2.25 g, 4.68 mmol) was dissolved in
DMSO (15 mL), and the solution was treated with 2 N
potassium hydroxide (2.5 mL) and then stirred at 60 °C under
nitrogen for an hour. The mixture was cooled to room
temperature, treated with hydrazine hydrate (0.4 mL, 11.7
mmol), and then heated at 60 °C for an additional hour.
Potassium tert-butoxide (1.31 g, 11.7 mmol) was added, and
the temperature was increased to 100 °C. After 18 h the
mixture was cooled, mixed with water, and acidified to pH 4
with 2 N HCl. The mixture was saturated with sodium
chloride and extracted with THF/ethyl acetate (1:1), and the
extracts were combined, dried (MgSO4), and then concentrated
under reduced pressure which gave 2.29 g of a dark oil. IR
1737, 1650 cm-1
.
Step 2. 1,4-Bis[3-(3-carbethoxymethylphenyl)-4-methoxy-
benzoyl]benzene (11a , 0.86 g, 1.28 mmol) was dissolved in 1,2-
dichloroethane (10 mL) under nitrogen, and chilled in an ice
bath. Boron tribromide (0.85 mL, 9.0 mmol) was added slowly,
then the cooling bath was removed, and the mixture was
stirred for 15 min. The reaction was cooled to 0 °C in an ice
bath, quenched with ethanol, and then mixed with ice water.
The mixture was extracted with dichloromethane, and the
extracts were combined, dried (MgSO4), and concentrated
under reduced pressure which provided 0.66 g (80%) of 1,4-
bis[3-(3-carboethoxymethylphenyl)-4-hydroxybenzoyl]ben-
zene (12a ). 1H NMR (400 MHz, CDCl3): 7.70-7.82 (m, 10H),
7.30-7.50 (m, 6H), 7.02-7.08 (m, 2H), 7.02 (s, 2H), 4.15 (q, J
) 7.6 Hz, 4H), 3.68 (s, 4H), 1.25 (t, J ) 7.6 Hz, 6H) ppm. IR
(NaCl): 1709 cm-1
. The crude product was dissolved in
dichloromethane (25 mL) and cooled in a dry ice/2-propanol
bath. Boron tribromide (2.4 mL, 25 mmol) was added drop-
wise, and the mixture was stirred at -78 °C for 2 h, warmed
to 0 °C for 2 h, then re-cooled to -78 °C, and quenched with
methanol. The mixture was concentrated under reduced
pressure, and the residue was partitioned between THF and
saturated sodium chloride. The organic material was sepa-
rated, dried (MgSO4), and then concentrated. The residue
(2.63 g) was dissolved in methanol (30 mL), and 5 drops of
concentrated sulfuric acid was added. The mixture was
refluxed overnight, filtered through a pad of sodium carbonate,
and then concentrated. Purification by flash chromatography
(SiO2, gradient elution, hexane to 3:1 hexane/ethyl acetate)
gave 0.89 g (43%) of methyl 3-(2-hydroxy-5-(1-oxo-2-(3-(2-
hydroxyphenyl)phenyl)ethyl)phenyl)phenylacetate 8 as a clear
oil. 1H NMR (400 MHz, CDCl3): 7.15-7.50 (m, 10H), 6.80-
7.10 (m, 5H), 5.26 (s, 1H), 5.17 (s, 1H), 3.71 (s, 3H), 3.68 (s,
(NaCl): 3352, 1732 cm-1
.
Step 3. 1,4-Bis[3-(3-carbethoxymethylphenyl)-4-hydroxy-
benzoyl]benzene (12a , 0.30 g, 0.46 mmol) and R-D-mannose
pentaacetate (0.54 g, 1.37 mmol) were dissolved in 1,2-
dichloroethane (5 mL), and then boron trifluoride etherate
(0.68 mL, 5.5 mmol) was added slowly. The mixture was
stirred under nitrogen for 16 h at room temperature and then
mixed with water. The organic material was separated, and
the aqueous portion was extracted with dichloromethane. The
combined organic fractions were dried (MgSO4) and then
concentrated under reduced pressure. The residue was dis-
solved in acetonitrile (7 mL), treated with 2 N sodium
hydroxide (9.8 mL), stirred at room temperature overnight,
and then acidified to pH 2 with concentrated HCl. The
volatiles were removed under reduced pressure, and a portion
of the residue was purified by reverse-phase HPLC (0-50%
acetonitrile in water) which gave 1,4-bis[3-(3-carboxymeth-
ylphenyl)-4-(R-D-mannopyranosyloxy)benzoyl]benzene (13a ) as
a white solid, mp 127-130 °C. 1H NMR (400 MHz, DMSO-
d6): 7.65-7.92 (m, 8H), 7.20-7.51 (m, 8H), 7.11 (d, J ) 8 Hz,
2H), 5.28 (s, 2H), 5.07 (br, 2H), 4.86 (br, 2H), 4.72 (br, 2H),
4.50 (br, 2H), 3.89 (s, 2H), 3.69-3.78 (m, 4H), 3.55-3.67 (m,
2H), 2.90-3.00 (m, 4H) ppm. IR (NaCl): 3417, 1718 cm-1
.
Step 4. The bis-phenol (8, 0.69 g, 1.57 mmol) was dissolved
in 1,2-dichloroethane (10 mL), and R-D-mannose pentaacetate
(1.8 g, 4.5 mmol) was added in one portion, and then boron
trifluoride etherate (2.3 mL, 18.8 mmol) was added slowly. The
mixture was stirred overnight at room temperature and then
mixed with water. The organic material was separated, and
the aqueous portion was extracted with dichloromethane. The
extracts were combined with the original organic fraction,
dried (MgSO4) then concentrated under reduced pressure.
Purification by flash chromatography (SiO2, gradient elution,
hexane to 3:1 hexane/ethyl acetate) provided 1.38 g (66%) of
the protected bis-glycoside as a foam, which was used in the
next step without further purification.
The product was dissolved in acetonitrile (5 mL), treated
with a solution of lithium hydroxide (0.24 g, 5.6 mmol in 5
mL water), stirred at room temperature overnight, then
acidified to pH 3 with concentrated HCl, and concentrated
under reduced pressure. The residue was purified by reverse-
phase HPLC (0-50% acetonitrile in water) which gave 3-(2-
R-D-mannopyranosyloxy-5-(2-(3-(2-R-D-mannopyranosyloxyphe-
nyl)phenyl)ethyl)phenyl)phenylacetic acid (9) (142 mg, 18%)
as a white solid, mp 129-134 °C. 1H NMR (400 MHz, DMSO-
d6): 7.10-7.40 (m, 15H), 5.33 (d, J ) 1.7 Hz, 1H), 5.27 (d, J )
1.7 Hz, 1H), 3.30-3.70 (m, 14H, plus H2O), 2.92 (2, 4H). IR
(KBr): 3410, 1710. MS (CI): m/e 425 [M - (mannose)2]+. Anal.
(C40H44O14‚2.2H2O) C, H.
4,4′-Bis(4-r-D-m a n n op yr a n osyloxyp h en yl)va ler ic a cid
(10). The compound was prepared from 4,4′-bis(4-hydrox-
yphenyl)valeric acid by glycosylation followed by base hydroly-
sis as described in the other examples: mp 130-132 °C. 1H
NMR (400 MHz, DMSO-d6): 7.08 (d, J ) 8.0 Hz, 4H), 6.98 (d,
J ) 8.0 Hz, 4H), 5.30 (s, 2H), 4.40 (br s, 8H), 3.79 (m, 2H),
3.65 (dd, J ) 3.3, 9.2 Hz, 2H), 3.59 (d, J ) 9.8 Hz, 2H), 3.38-
8H) ppm. IR (KBr): 3424, 1716, 1642, 1594 cm-1
. Anal.
(C48H44O16‚0.6TFA) C, H.
1,6-Bis[3-(3-ca r boxym eth ylp h en yl)-4-(r-D-m a n n op yr a -
n osyloxy)p h en yl]h exa n e (15e): Step 1. Adipoyl chloride
(2.0 g, 10.9 mmol) was dissolved in 1,2-dichloroethane (55 mL)
and cooled in an ice bath. Aluminum chloride (5.8 g, 43.5
mmol) was added followed by methyl 3-(2-methoxyphenyl)-
phenylacetate (4, 11.2 g, 43.5 mmol), and the mixture was
stirred at room temperature overnight. The mixture was
cooled in an ice bath, and ice water was added slowly while
stirring. The organic materials were isolated, and the aqueous
portion was extracted with dichloromethane. The combined
organic materials were dried (MgSO4) and then concentrated
under reduced pressure. Purification by flash chromatography
(SiO2, gradient elution, hexane to 3:1 hexane/ethyl acetate)
provided 2.23 g (33%) of 11e. 1H NMR (400 MHz, CDCl3): 7.96
(dd, J ) 6.6, 1.9 Hz, 2H), 7.92 (d, J ) 1.9 Hz, 2H), 7.42 (m,
2H), 7.34 (t, J ) 6 Hz, 2H), 7.18-7.28 (m, 4H), 7.00 (d, J )

Inhibitors of Selectin-Mediated Cell Adhesion
J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 7 1107
6.3 Hz, 2H), 3.87 (s, 6H), 3.69 (s, 6H), 3.68 (s, 4H), 3.01 (m,
romethane, and the combined extracts were washed with
saturated sodium chloride, dried (MgSO4), and then concen-
trated under reduced pressure. Purification through silica gel
with 10:1 hexane/ethyl acetate gave 2.49 g (82%) of 4-(4-
methoxyphenyl)butyryl-4-methoxybenzene. 1H NMR (400 MHz,
CDCl3): 7.90 (d, J ) 8.8 Hz, 2H), 7.11 (d, J ) 8.4 Hz, 2H),
6,91 (d, J ) 8.8 Hz, 2H), 6.83 (d, J ) 8.4 Hz, 2H), 3.86 (s, 3H),
3.78 (s, 3H), 2.90 (t, J ) 7.3 Hz, 2H), 2.65 (t, J ) 7.7 Hz, 2H),
4H), 1.84 (m, 4H) ppm. IR (NaCl): 1741, 1677 cm-1
.
Step 2. The product from step 1 (2.23 g, 3.6 mmol) was
dissolved in acetonitrile, treated with a solution of lithium
hydroxide (0.76 g, 18 mmol in 8 mL water), stirred at room
temperature overnight, acidified to pH 4 with 2 N HCl, and
extracted with ethyl acetate. The extracts were combined,
dried (MgSO4), and then concentrated under reduced pressure.
The keto acid (1.86 g, 3.1 mmol) was dissolved in dimethyl
sulfoxide (15 mL), hydrazine (1.0 mL, 31 mmol) was added
and the mixture was heated at 80 °C under nitrogen for 2.5 h.
After cooling, potassium tert-butoxide (3.5 g, 31 mmol) was
added, and the mixture was again heated at 80 °C overnight,
mixed with water, acidified with 2 N HCl, and extracted with
ethyl acetate. The extracts were combined, dried (MgSO4), and
concentrated under reduced pressure which gave 1.6 g (91%)
of the diacid.
The diacid (1.6 g, 2.8 mmol) was dissolved in dichlo-
romethane (14 mL) under nitrogen and chilled to -78 °C.
Boron tribromide (1.4 mL, 14 mmol) was added slowly, and
the mixture was then stirred at room temperature for 2 h and
subsequently quenched with ice water. The organic material
was separated, washed with saturated sodium chloride, then
dried (MgSO4), and concentrated under reduced pressure
which gave 2.38 g of the crude phenol.
2.02 (m, 2H) ppm. IR (NaCl): 1674, 1602, 1244 cm-1
.
Step 2. The ketone (2.49 g, 8.8 mmol) was dissolved in
dichloromethane (30 mL) and treated with trifluoroacetic acid
(2.7 mL, 35.2 mmol), triethylsilane (2.8 mL, 17.6 mmol), and
then boron trifluoride etherate (4.4 mL, 35.2 mmol). The
mixture was stirred at room temperature for 2 h, then cooled
in an ice bath, and mixed with water. The mixture was
extracted with dichloromethane, and the combined extracts
were washed with saturated sodium chloride, dried (MgSO4),
and then concentrated under reduced pressure. The residue
was flushed through silica gel with 10:1 hexane/ethyl acetate
and concentrated which provided 2.0 g (85%) of 17 as a clear
oil. 1H NMR (400 MHz, CDCl3): 7.07 (d, J ) 8.4 Hz, 4H), 6.81
(d, J ) 8.4 Hz, 4H), 3.78 (s, 6H), 2.56 (m, 4H), 1.61 (m, 4H)
ppm. IR (NaCl): 1605, 1248 cm-1
.
Step 3. The bis-ether 17 (1.78 g, 6.6 mmol) and TMEDA
(4.0 mL, 26.5 mmol) were mixed with anhydrous ether (30 mL)
and chilled in an ice bath. n-Butyllithium (10.5 mL of a 2.5
M solution, 26.5 mmol) was added, and the mixture was
warmed to room temperature and stirred for an hour. The
reaction mixture was cooled to 0 °C and treated with trimethyl
borate (3.0 mL, 26.5 mmol). The mixture was stirred at room
temperature overnight, then quenched with 2 N HCl (to pH
2), and stirred for 1 h. The organic phase was separated, the
aqueous portion was extracted with ethyl acetate, and the
extracts were combined with the original organic fraction,
dried (MgSO4), and then concentrated under reduced pressure,
which gave 3.0 g of the boronic acid which was used without
The residue from the previous step was mixed with metha-
nol (50 mL), and sulfuric acid (5 drops) was added. The
mixture was heated at reflux overnight and then concentrated
under reduced pressure. The residue was dissolved in dichlo-
romethane (50 mL), treated with sodium carbonate, and then
filtered through a pad of silica gel. The filtrate was concen-
trated under reduced pressure, and the residue was purified
by flash chromatography (SiO2, gradient elution, hexane to 3:1
hexane/ethyl acetate) which provided 0.9 g (38%) of 1,6-bis-
[3-(3-carbomethoxymethylphenyl)-4-hydroxyphenyl]hexane
(14e). 1H NMR (400 MHz, CDCl3): 6.80-7.50 (m, 14H), 3.70
(s, 6H), 3.68 (s, 4H), 2.55 (dd, J ) 5.5, 5.5 Hz, 4H), 1.59 (m,
further purification. IR (NaCl): 1328 cm-1
.
4H), 1.36 (m, 4H) ppm. IR (NaCl): 3430, 1731 cm-1
.
A solution of the crude boronic acid and methyl 3-bromo-
phenylacetate (3.8 g, 16.8 mmol) in dimethoxyethane (30 mL)
was degassed under nitrogen. The mixture was treated with
tribasic potassium phosphate (10.7 g, 50.5 mmol) and bis-
(triphenylphosphine)palladium(II) chloride (100 mg, 0.17 mmol).
The mixture was degassed again, then heated at reflux for 2
h, cooled to room temperature, and mixed with water. The
mixture was extracted with dichloromethane, and the com-
bined extracts were washed with water and saturated sodium
chloride, dried (MgSO4), and then concentrated under reduced
pressure. Purification by flash chromatography (SiO2, gradi-
ent elution, 8:1 hexane/ethyl acetate to 4:1 hexane/ethyl
acetate) gave 1.14 g (24%) of 18 as a clear oil. 1H NMR (400
MHz, CDCl3): 7.41-7.44 (m, 4H), 7.34 (t, J ) 7.7 Hz, 2H),
7.21-7.60 (m, 2H), 7.08-7.13 (m, 4H), 6.87 (d, J ) 8.0 Hz,
2H), 3.77 (s, 6H), 3.68 (s, 6H), 3.66 (s, 4H), 2.61 (m, 4H), 1.67
Step 3. The bisphenol (14e, 0.9 g, 1.6 mmol) was dissolved
in 1,2-dichloroethane (8 mL) and glycosylated with R-D-
mannose pentaacetate (1.9 g, 4.8 mmol) as before, which
provided 1.5 g (77%) of 1,6-bis[3-(3-carbomethoxymethylphen-
yl)-4-(2,3,4,6-tetra-O-acetyl-R-D-mannopyranosyloxy)phenyl]-
hexane contaminated with a small amount of unreacted R-D-
mannose pentaacetate which coeluted with the product. 1H
NMR (400 MHz, CDCl3): 7.31-7.37 (m, 6H), 7.19-7.24 (m,
4H), 7.09-7.13 (m, 4H), 5.25 (d, J ) 0.6 Hz, 2H), 3.57-3.66
(m, 6H), 3.3-3.50 (m, 10H), 2.54 (m, 4H), 1.58 (m, 4H), 1.34
(m, 4H) ppm. IR (NaCl): 1752 cm-1
.
1,6-Bis[3-(3-carbomethoxymethylphenyl)-4-(2,3,4,6-tetra-O-
acetyl-R-D-mannopyranosyloxy)phenyl]hexane (1.5 g, 1.2 mmol)
hydrolyzed with lithium hydroxide solution as before. Puri-
fication by reverse-phase HPLC (5-50% acetonitrile in water)
gave 0.35 g (33%) of 1,6-bis[3-(3-carboxymethylphenyl)-4-(2-
R-D-mannopyranosyloxy)phenyl] hexane (15e) as a white solid,
mp 115-117 °C. 1H NMR (400 MHz, DMSO-d6): 7.31-7.37
(m, 6H), 7.19-7.24 (m, 4H), 7.09-7.13 (m, 4H), 5.25 (d, J )
0.6 Hz, 2H), 3.57-3.66 (m, 6H), 3.3-3.50 (m, 10H), 2.54 (m,
4H), 1.58 (m, 4H), 1.34 (m, 4H) ppm. IR (KBr): 3420, 1711
cm-1. MS (CI): m/e 864 (M + H)+, 683 (M - mannose)+. Anal.
(C46H54O16‚0.75TFA) C, H.
(m, 4H) ppm. IR (NaCl): 1737, 1606, 1239 cm-1
.
Step 4. 1,4-Bis[3-(3-carbomethoxymethylphenyl)-4-meth-
oxyphenyl]butane (18, 0.9 g, 1.6 mmol) was dissolved in
dichloromethane (3.0 mL), cooled to -78 °C, and treated with
boron tribromide (1.2 mL, 12.8 mmol) as before which gave
0.4 g (47%) of 1,4-bis[3-(3-carbomethoxymethylphenyl)-4-hy-
droxyphenyl]butane. 1H NMR (400 MHz, CDCl3): 7.42 (d, J
) 7.4 Hz, 2H), 7.35-7.38 (m, 4H), 7.27-7.32 (m, 2H), 7.02-
7.05 (m, 4H), 6.87 (d, J ) 8.0 Hz, 2H), 5.12 (s, 2H), 3.70 (s,
6H), 3.68 (s, 4H), 2.59 (m, 4H), 1.65 (m, 4H) ppm. IR (NaCl):
1,4-Bis[3-(3-ca r boxym eth ylp h en yl)-4-(2-r-D-m a n n op y-
r a n osyloxy)p h en yl] bu ta n e (19): Step 1. 4-(4-Methox-
yphenyl)butyric acid (2.0 g, 10.3 mmol) was treated with
thionyl chloride (20 mL), stirred at room temperature for 3 h,
then heated at 65 °C overnight, and concentrated under
reduced pressure which gave 2.3 g of 4-(4-methoxyphenyl)-
butyryl chloride as a yellow oil which was used without further
3439, 1732 cm-1
.
Step 5. Glycosylation of 1,4-bis[3-(3-carbomethoxymeth-
ylphenyl)-4-hydroxyphenyl]butane (0.4 g, 0.74 mmol) with R-D-
mannose pentaacetate (0.72 g, 1.85 mmol) in 1,2-dichloroeth-
ane (4 mL) as before gave 0.88 g (99%) of 1,4-bis[3-(3-
carbomethoxymethylphenyl)-4-(tetra-O-acetyl-R-D-manno-
pyranosyloxy)phenyl]butane as a foam. 1H NMR (400 MHz,
CDCl3): 7.38-7.44 (m, 6H), 7.25-7.29 (m, 2H), 7.17 (br s, 2H),
7.07-7.11 (m, 4H), 5.40 (s, 2H), 5.20-5.30 (m, 6H), 4.15 (dd,
J ) 12.3, 4.8 Hz, 2H), 3.93 (dd, J ) 12.4, 2.2 Hz, 2H), 3.76-
3.82 (m, 2H), 3.72 (s, 4H), 3.68 (s, 6H), 2.63 (m, 4H), 2.13 (s,
purification. IR (NaCl): 1795 cm-1
.
The crude acid chloride (2.07 g, 9.7 mmol) and anisole (1.26
g, 11.6 mmol) were dissolved in 1,2-dichloroethane (32 mL)
and chilled in an ice bath. Aluminum chloride (3.9 g, 29.1
mmol) was added in portions, stirred for 5 min, and then mixed
with ice water. The mixture was extracted with dichlo-

1108 J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 7
Kogan et al.
6H), 2.01 (s, 6H), 2.00 (s, 6H), 1.97 (s, 6H), 1.67 (m, 4H) ppm.
prop-1-ylthio]propane (0.47 g, 0.68 mmol) was glycosylated
with R-D-mannose pentaacetate (0.8 g, 2.0 mmol) as before,
which gave 0.331 g (36%) of 1,3-bis[3-(3-(3-carbethoxymeth-
ylphenyl)-4-(2,3,4,6-tetra-O-acetyl-R-D-mannopyranosyloxy)-
phenyl)prop-1-ylthio]propane 21 contaminated with a small
amount of unreacted R-D-mannose pentaacetate which coeluted
IR (NaCl): 1748, 1219 cm-1
.
Step 6. 1,4-Bis[3-(3-carbomethoxymethylphenyl)-4-(tetra-
O-acetyl-R-D-mannopyranosyloxy)phenyl]butane (0.87 g, 0.73
mmol) was dissolved in acetonitrile (3 mL) and hydrolyzed with
a solution of lithium hydroxide as before which gave 230 mg
of 19 as a white solid, mp 195-197 °C. 1H NMR (400 MHz,
DMSO-d6): 7.31-7.39 (m, 6H), 7.20-7.25 (m, 4H), 7.10-7.15
(m, 4H), 5.25 (s, 2H), 4.88 (br d, J ) 4.0 Hz, 2H), 4.76 (br s,
2H), 4.60 (br s, 2H), 4.45 (br t, J ) 5.9 Hz, 2H), 3.55-3.68 (m,
5H), 3.62 (s, 4H), 3.40-3.50 (m, 7H), 2.60 (m, 4H), 1.62 (m,
with the product. IR (NaCl): 1752 cm-1
.
Step 4. The tetraacetate 21 (0.64 g, 0.47 mmol) dissolved
in acetonitrile (5 mL) was treated with 2 N sodium hydroxide
(5.0 mL) and stirred overnight at room temperature. The
mixture was acidified with concentrated hydrochloric acid to
pH 2, and the volatiles were removed under reduced pressure.
A portion of the aqueous residue was purified by reverse-phase
HPLC (10-50% acetonitrile in water) which provided 22 as a
white solid, mp 96-100 °C. 1H NMR (400 MHz, DMSO-d6):
7.31-7.39 (m, 6H), 7.20-7.27 (m, 4H), 7.11-7.15 (m, 4H), 5.26
(s, 2H), 3.30-3.75 (m, 28H), 2.64 (t, J ) 7.7 Hz, 4H), 2.57 (t,
J ) 7.0 Hz, 4H), 2.45-2.52 (m, 4H), 1.75-1.84 (m, 4H), 1.66-
4H) ppm. IR (KBr): 3333, 3229, 1729, 1224 cm-1
. Anal.
(C44H50O16‚0.4TFA) C, H.
1,3-Bis[3-(3-(3-ca r boxym eth ylp h en yl)-4-(r-D-m a n n op y-
r a n osyloxy)p h en yl)p r op -1-ylth io]p r op a n e (22). Step 1.
Ethyl 3-(2-methoxyphenyl)phenyl acetate (5, 5.04 g, 18.66
mmol) and 3-bromopropionyl chloride (1.88 mL, 18.66 mmol)
were mixed with 1,2-dichloroethane (30 mL), and the mixture
was cooled to 0 °C and then treated with aluminum chloride
(7.6 g, 57 mmol). After 15 min the reaction was mixed with
ice water, and the organic materials were separated. The
aqueous portion was extracted with dichloromethane, and the
organic materials were combined, dried (MgSO4), and then
concentrated under reduced pressure, which gave 11i.
1.74 (m, 2H). IR (KBr): 3430, 1711 cm-1. Anal. (C49H60S2O16
0.5TFA) C, H.
‚
1-(3-(3-Ca r b oxym e t h ylp h e n yl)-4-r-D-m a n n op yr a n o-
syloxyp h en yl)-6-(4-r-D-m a n n op yr a n osyloxyp h en yl)h ex-
a n e (26). Step 1. Adipic acid monomethyl ester (2.0 g, 12.5
mM) was treated with thionyl chloride (1.1 mL, 15 mM), and
the mixture was heated to 80 °C for 30 min and then
concentrated under reduced pressure. The residue was mixed
with dichloromethane (20 mL) and cooled to 0 °C. Anisole
(1.36 mL, 12.5 mM) was added and then aluminum chloride
(4.25 g, 37.5 mM). After 30 min, the reaction was mixed with
ice water and extracted with dichloromethane. The extracts
were dried (MgSO4) and concentrated under reduced pressure.
1H NMR (400 MHz, CDCl3): 7.92 (d, J ) 12 Hz, 2H), 6.92 (d,
J ) 12 Hz, 2H), 3.86 (s, 3H), 3.66 (s, 3H), 2.93 (t, J ) 6.8 Hz,
2H), 2.36 (t, J ) 6.8 Hz, 2H), 1.62-1.79 (m, 4H) ppm. IR
Step 2. The bromo ketone 11i (8.5 g, 19 mmol) was
dissolved in dichloromethane (40 mL) and cooled in an ice
water bath. Trifluoroacetic acid (5.9 mL, 76 mmol), triethyl-
silane (6.1 mL, 38 mmol), and then boron trifluoride etherate
(9.4 mL, 76 mmol) were added, and the cooling bath was
removed. The mixture was stirred at room temperature
overnight, then cooled in an ice bath, and quenched with cold
water. The organic materials were separated, the aqueous
portion was extracted with dichloromethane, and the organic
materials were combined, washed with saturated sodium
chloride solution, dried (MgSO4), and then concentrated under
reduced pressure, which provided 6.53 g (90%) of ethyl 3-(2-
methoxy-5-(3-bromopropyl)phenyl)phenylacetate 20. 1H NMR
(400 MHz, CDCl3): 7.41-7.45 (m, 2H), 7.36 (t, J ) 8 Hz, 1H),
7.25-7.28 (m, 1H), 7.12-7.16 (m, 2H), 6.90 (d, J ) 8.6 Hz,
1H), 4.15 (q, J ) 7.0 Hz, 2H), 3.78 (s, 3H), 3.67 (s, 2H), 3.41 (t,
J ) 6.6 Hz, 2H), 2.75 (t, J ) 7.0 Hz, 2H), 2.15 (m, 2H), 1.26 (t,
(NaCl): 3433, 1731, 1675 cm-1
. A portion of the keto ester
(1.0 g, 4.0 mM) was dissolved in THF (6 mL) and treated with
2 N sodium hydroxide (2.25 mL), and the mixture was stirred
at room temperature overnight and then neutralized with
concentrated hydrochloric acid (pH 5.5). The solids were
collected and dried under vacuum which gave 0.78 g (87%) of
5-(4-methoxybenzoyl)valeric acid. 1H NMR (400 MHz,
DMSO-d6): 7.94 (d, J ) 12 Hz, 2H), 7.03 (d, J ) 12 Hz, 2H),
3.84 (s, 3H), 2.96 (t, J ) 6.8 Hz, 2H), 2.24 (t, J ) 6.8 Hz, 2H),
J ) 7.3 Hz, 3H). IR (NaCl): 1736 cm-1
.
Step 3. A solution of 1,3-propanedithiol (0.109 g, 0.94
mmol) in THF (4.5 mL) was degassed and then cooled to 0 °C.
Sodium hydride (86.5 mg, 2.1 mmol) was added, and the
mixture was stirred at room temperature for 2 h. A solution
of the bromide 20 (0.83 g, 2.12 mmol) in THF (1.0 mL) was
added, and the mixture was heated under reflux overnight.
The reaction was partitioned between water and ethyl acetate,
and the organic materials were separated, washed with
saturated sodium chloride, dried (MgSO4), and then concen-
trated under reduced pressure. Purification by flash chroma-
tography (SiO2, gradient elution, hexane to 3:1 hexane/ethyl
acetate) gave 166.2 mg (24%) of 1,3-bis[3-(3-(3-carbethoxy-
methylphenyl)-4-methoxyphenyl)prop-1-ylthio]propane. 1H
NMR (400 MHz, CDCl3): 7.39-7.45 (m, 4H), 7.32-7.38 (m,
2H), 7.21-7.28 (m, 2H), 7.08-7.15 (m, 4H), 6.85-6.92 (m, 2H),
4.14 (q, J ) 7.0 Hz, 4H), 3.77 (s, 6H), 3.65 (s, 4H), 2.80-2.95
(m, 2H), 2.77 (t, J ) 7.3 Hz, 2H), 2.68 (t, J ) 8.0 Hz, 4H), 2.60
(t, J ) 7.0 Hz, 2H), 2.52 (t, J ) 7.0 Hz, 2H), 2.04-2.16 (m,
2H), 1.81-1.93 (m, 4H), 1.25 (t, J ) 7.0 Hz, 6H). IR (NaCl):
1.50-1.70 (m, 4H) ppm. IR (NaCl): 3472, 1702, 1668 cm-1
.
Step 2. 5-(4-Methoxybenzoyl)valeric acid (0.7 g, 2.96 mM)
was mixed with thionyl chloride (0.43 mL, 5.9 mM), heated at
80 °C for 10 min, and then concentrated under reduced
pressure. The residue was dissolved in dichloromethane (10
mL), and ethyl 3-(2-methoxyphenyl)phenylacetate (5, 0.81 g,
3.0 mM) and then aluminum chloride (2.0 g, 15 mM) were
added. The mixture was stirred at room temperature over-
night, then quenched with ice water, and extracted with
dichloromethane. The extracts were dried (MgSO4), filtered
through a pad of silica gel, and concentrated under reduced
pressure, which gave 1.1 g (79%) of 11j as a clear oil, which
was used without further purification. IR (NaCl): 1730, 1666,
1598 cm-1
. The diketone was dissolved in dichloromethane
(12 mL) and treated with TFA (1.8 mL, 23.1 mM), boron
trifluoride etherate (2.9 mL, 23.1 mM), and then triethylsilane
(1.8 mL, 11.5 mM). The mixture was stirred at room temper-
ature for 10 h and then mixed with water. The mixture was
extracted with dichloromethane, and the extracts were dried
(MgSO4) and then concentrated under reduced pressure which
gave 0.61 g (59%) of the bis-ether 24. IR (NaCl): 1732, 1172
1731 cm-1
.
A solution of the bis-sulfide (70.7 mg, 0.1 mmol) in dichlo-
romethane (2 mL) was treated with boron tribromide (0.8 mL
of 1 M in dichloromethane, 0.8 mmol) as before, which gave
68 mg (100%) of 1,3-bis[3-(3-(3-carbethoxymethylphenyl)-4-
hydroxyphenyl)prop-1-ylthio]propane. 1H NMR (400 MHz,
CDCl3): 7.43 (t, J ) 7.7 Hz, 2H), 7.34-7.39 (m, 4H), 7.30 (br
d, J ) 7.3 Hz, 2H), 7.03-7.08 (m, 4H), 6.87 (br d, J ) 8.8 Hz,
2H), 4.96-5.30 (br s, 2H), 4.15 (q, J ) 7.0 Hz, 4H), 3.66 (s,
4H), 2.66 (t, J ) 7.3 Hz, 4H), 2.60 (t, J ) 7.0 Hz, 2H), 2.52 (t,
J ) 7.0 Hz, 4H), 1.80-1.92 (m, 6H), 1.26 (t, J ) 7.3 Hz, 6H),
cm-1
.
Step 3. The bis-ether 24 was dissolved in dichloromethane
(6 mL), cooled to -78 °C, and treated with boron tribromide
(1.1 mL, 12 mM). The reaction mixture then was stirred for
3 h at 0 °C and quenched with ethanol. Water was added,
and the mixture was extracted with dichloromethane. The
extracts were dried (MgSO4) and then concentrasted under
reduced pressure. Purification by flash chromatography (SiO2,
gradient elution, hexane to 3:1 hexane/ethyl acetate) gave 100
mg (17%) of the di-phenol 25. 1H NMR (400 MHz, DMSO-d6):
1.21-1.27 (m, 2H). IR (NaCl): 3420, 1726 cm-1
1,3-Bis[3-(3-(3-carbethoxymethylphenyl)-4-hydroxyphenyl)-
.

Inhibitors of Selectin-Mediated Cell Adhesion
J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 7 1109
7.28-7.45 (m, 4H), 6.97-7.52 (m, 4H), 6.87 (d, J ) 8.0 Hz,
1H), 6.69 (d, J ) 8 Hz, 2H), 4.16 (q, J ) 8.2 Hz, 2H), 3.67 (s,
3H), 2.51 (m, 4H), 1.58 (m, 4H), 1.35 (m, 4H) ppm.
4.70 (br s, 3H), 4.50 (br s, 3H), 3.88-3.92 (m, 6H), 3.58-3.73
(m, 18H), 3.40-3.49 (m, 6H), 3.30-3.40 (m, 6H plus H2O). IR
(KBr): 3429, 1708, 1669 cm-1
.
1,3-Bis[N-(4-(3-(3-ca r b oxym et h ylp h en yl)-4-(r-D-m a n -
n op yr a n osyloxy)p h en yl)b u t a n oyl)p ip er id in -4-yl]p r o-
p a n e (37b). Step 1. Succinic anhydride (2.0 g, 19.9 mmol)
and aluminum chloride (17.7 g, 132 mmol) in 1,2-dichloroeth-
ane (45 mL) were cooled to 0 °C. The mixture was treated
with a solution of ethyl 3-(2,3,4,6-tetra-O-pivaloyl-R-D-man-
nopyranosyloxyphenyl)phenylacetate16 34 (10.0 g, 13.2 mmol)
in 1,2-dichloroethane (10 mL), and the mixture was stirred
overnight while the bath gradually came to room temperature.
The reaction was quenched with ice water and stirred for 15
min. The organic materials were isolated, and the aqueous
portion was extracted with dichloromethane. The organic
materials were combined, dried (MgSO4), and then concen-
trated under reduced pressure, which gave 13.5 g of 35 which
was used without further purification. IR (NaCl): 1738, 1685
Step 4. The di-phenol (100 mg, 0.23 mM) was dissolved in
dichloromethane (2 mL) and treated with R-D-mannnose
pentaacetate (0.25 g, 0.65 mM) and then boron trifluoride
etherate (0.26 mL, 2.1 mM). After 16 h, the reaction was
quenched with water, and the organic material was separated,
dried (MgSO4), and concentrated under reduced pressure. The
residue was dissolved in acetonitrile (2 mL), and 2 N sodium
hydroxide (1.2 mL) was added. After 18 h, the mixture was
acidified with concentrated HCl (pH 3). Purification of a
portion by reverse-phase HPLC (10-100% acetonitrile in
water) provided 22 mg of 26 as a white solid, mp 106-108 °C.
1H NMR (400 MHz, DMSO-d6): 7.34-7.40 (m, 3H), 7.21-7.25
(m, 2H), 7.06-7.13 (m, 4H), 6.96 (d, J ) 8 Hz, 2H), 5.28 (d, J
) 1.5 Hz, 1H), 5.25 (d, J ) 1.5 Hz, 1H), 3.30-3.81 (m, 14H),
2.55 (m, 4H), 1.55 (m, 4H), 1.33 (m, 4H) ppm. IR (NaCl): 3400,
1734 cm-1. Anal. (C38H48O14‚0.5TFA) C, H.
cm-1
.
Step 2. To a solution of 35 (13.5 g, 19.7 mmol) in
dichloromethane (65 mL) was added boron trifluoride etherate
(15.3 mL, 122 mmol), trifluoroacetic acid (9.4 mL, 122 mmol),
and then triethylsilane (9.4 mL, 59.1 mmol), and the mixture
was stirred at room temperature overnight. The reaction was
quenched with water (200 mL), and the organic materials were
separated. The aqueous portion was extracted with dichlo-
romethane, and the extracts were combined with the original
organic portion, dried (MgSO4), and then concentrated under
reduced pressure which provided 1.49 g (97%) of 4-(3-(3-
carbethoxymethylphenyl)-4-(2,3,4,6-tetra-O-pivaloyl-R-D-man-
1,3,5-Tr is[(3-(3-car boxym eth ylph en yl)-4-(r-D-m an n opy-
r a n osyloxy)p h en yl)-2-oxoeth ylth iom eth yl]ben zen e (33).
Step 1. Ethyl 3-(2,3,4,6-tetra-O-acetyl-R-D-mannopyranosyl-
oxyphenyl)phenylacetate 27 (0.32 g, 0.547 mmol) and bro-
moacetyl bromide (0.06 mL, 0.689 mmol) were dissolved in 1,2-
dichloroethane (3 mL) and cooled to -78 °C. The mixture was
treated with aluminum chloride (1.45 g, 10.9 mmol), and the
bath was replaced with an ice water bath. After 15 min, the
reaction was quenched with ice water and stirred for 15 min.
The organic materials were isolated, and the aqueous portion
was extracted with dichloromethane. The organic materials
were combined, dried (MgSO4), and then concentrated under
reduced pressure, which gave 0.387 g of 30, which was used
without further purification. 1H NMR (400 MHz, CDCl3): 8.03
(d, J ) 2.5 Hz, 1H), 7.95 (dd, J ) 8.8, 2.5 Hz, 1H), 7.41-7.47
(m, 3H), 7.35 (m, 1H), 7.31 (d, J ) 8.8 Hz, 1H), 5.62 (d, J )
1.8 Hz, 1H), 5.21-5.34 (m, 3H), 4.42 (s, 2H), 4.10-4.25 (m,
5H), 3.99 (dd, J ) 12.1, 2.2 Hz, 1H), 3.80-3.85 (m, 1H), 3.74
(s, 2H), 2.18 (s, 3H), 2.03 (s, 3H), 2.02 (s, 3H), 1.98 (s, 3H),
nopyranosyloxy)phenyl)butanoic acid as
a clear oil. IR
(NaCl): 1737 cm-1
.
Step 3. The acid prepared in step 2 (16.0 g, 23.8 mmol)
was stirred with thionyl chloride (50 mL), and the mixture
was stirred at room temperature for 36 h and then concen-
trated under reduced pressure, which gave 16.3 g (99%) of the
acid chloride, which was used without further purification. IR
(NaCl): 1797, 1740 cm-1. 1,3-Bis(piperidin-4-yl)propane (0.4
g, 1.9 mmol) in dichloromethane (5 mL) was added to a solution
of the acid chloride (2.8 g, 3.26 mmol) in dichloromethane (5
mL) at 0 °C. Triethylamine (0.61 mL, 4.4 mmol) and 4-(di-
methylamino)pyridine (35 mg, 10 mol %) were added, and the
reaction was stirred at room temperature for 1 h and then
quenched with water. The organic materials were washed
with saturated sodium chloride, then dried (MgSO4), and
concentrated under reduced pressure. Purification by flash
chromatography (SiO2, gradient elution, hexane to 3:1 hexane/
ethyl acetate) gave 1.1 g (18%) of the bis-amide 36b. 1H NMR
(400 MHz, CDCl3): 7.40-7.45 (m, 4H), 6.98-7.30 (m, 10 H),
5.25-5.42 (m, 6H), 4.09-4.19 (m, 4H), 3.50-3.96 (m, 8H),
2.62-2.69 (m, 4H), 2.28-2.36 (m, 4H), 1.90-2.02 (m, 4H),
1.60-1.74 (m, 6H), 1.35-1.50 (m, 2H), 1.08-1.28 (98H). IR
1.26 (t, J ) 8.0 Hz, 3H). IR (NaCl): 1746 1220 cm-1
.
Step 2. 1,3,5-Tris(mercaptomethyl)benzene (102 mg, 0.47
mmol) in THF (1 mL) was treated with sodium hydride (59.8
mg, 1.49 mmol), and the mixture was stirred at room temper-
ature for 30 min. A solution of bromo ketone 30 (1.02 g, 1.44
mmol) in THF (2.0 mL) was added, and the mixture was
stirred at room temperature overnight. The reaction was
mixed with water and extracted with ethyl acetate. The
organic materials were combined, dried (MgSO4), and concen-
trated under reduced pressure. Purification by flash chroma-
tography (SiO2, gradient elution, 3:1 hexane/ethyl acetate to
1:1 hexane/ethyl acetate) gave 0.59 g (60%) of the correspond-
ing trisulfide. 1H NMR (400 MHz, CDCl3): 7.98 (d, J ) 2.5
Hz, 3H), 7.89 (dd, J ) 8.8, 2.5 Hz, 3H), 7.40-7.46 (m, 6H),
7.25-7.35 (m, 12H), 5.61 (s, 3H), 5.22-5.33 (m, 9H), 4.10-
4.25 (m, 10H), 3.97 (dd, J ) 12.1, 2.2 Hz, 3H), 3.78-3.84 (m,
3H), 3.72 (s, 6H), 3.69 (s, 3H), 3.64 (s, 3H), 2.16 (s, 9H), 2.02
(s, 9H), 2.01 (s, 9H), 1.97 (s, 9H), 1.25 (t, J ) 8.0 Hz, 9H). IR
(NaCl): 1739 cm-1
.
Step 4. Bisamide 36b (1.0 g, 0.54 mmol) was dissolved in
THF (3 mL), 0.33 N sodium hydroxide (3 mL) was added, and
the mixture was stirred at room temperature overnight. The
THF was removed under reduced pressure, and the residue
was acidified to pH 2 with concentrated hydrochloric acid.
Purification by reverse-phase HPLC (20-80% acetonitrile in
water) gave 30 mg (5%) of 1,3-bis[N-(4-(3-(3-carboxymeth-
ylphenyl)-4-(R-D-mannopyranosyloxy)phenyl)butanoyl)piperidin-
4-yl]propane (37b) as a white solid, mp 119-121 °C. 1H NMR
(400 MHz, DMSO-d6): 7.32-7.40 (m, 6H), 7.20-7.27 (m, 4H),
7.10-7.14 (m, 4H), 5.26 (br s, 2H), 4.36 (br d, J ) 11.0 Hz,
2H), 4.05 (br s, 8H), 3.77 (br d, J ) 11.0 Hz, 2H), 3.50-3.70
(m, 8H), 3.40-3.52 (m, 5H), 3.31-3.40 (m, 2H), 2.92 (br t, J )
14.0 Hz, 2H), 2.55-2.62 (m, 4H), 2.42-2.53 (m, 3H), 2.27-
2.34 (m, 4H), 1.73-1.82 (m, 4H), 1.57-1.67 (m, 4H), 1.36-
1.47 (m, 2H), 1.22-1.33 (m, 2H), 1.11-1.20 (m, 4H), 0.80-
1.02 (m, 4H). IR (KBr): 3415, 1713, 1609 cm-1. MS (FAB):
1150 (M + Na)+. Anal. (C61H78O18‚1.4TFA) C,H.
(NaCl): 1747, 1219 cm-1
.
Step 3. The trisulfide from step 2 (0.59 g, 0.28 mmol) in
THF (3 mL) was treated with a solution of freshly prepared
sodium methoxide (93 mg, 4.04 mmol in 3 mL methanol), and
the mixture was stirred at room temperature overnight. The
precipitate which had formed was collected by vacuum filtra-
tion, washed several times with cold THF/methanol (2:1), and
dried under vacuum which gave 0.38 g of a white solid. The
solid was dissolved in water (12 mL), 2 N sodium hydroxide
was added to pH 14, and the mixture was stirred at room
temperature for 4 h. The reaction was acidified with Dowex
50W acidic ion-exchange resin and filtered, and the volatiles
were removed under reduced pressure. Purification of a
portion by reverse-phase HPLC (20-80% acetonitrile in water)
gave 135 mg of 33 as a white solid, mp 140-143 °C. 1H NMR
(400 MHz, DMSO-d6): 7.95 (dd, J ) 8.8, 2.5 Hz, 3H), 7.89 (d,
J ) 2.2 Hz, 3H), 7.32-7.45 (m, 12H), 7.26 (d, J ) 7.3 Hz, 3H),
7.16-7.20 (m, 3H), 5.54 (s, 3H), 5.05 (br s, 3H), 4.85 (br s, 3H),
1,6-Bis(3-(3-ca r boxym eth oxyp h en yl)-4-(r-D-m a n n op y-
r a n osyloxy)p h en yl) h exa n e (42). Step 1. Anisole (1.0 g,
9.25 mmol) was dissolved in dichloromethane (20 mL) and

1110 J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 7
Kogan et al.
cooled to 0 °C. Adipoyl chloride (0.7 mL, 4.62 mmol)and then
aluminum chloride (6.2 g, 46 mmol) were added in portions.
After 4 h, the reaction was quenched with ice water and
extracted with dichloromethane. The organic materials were
combined, dried (MgSO4), and then concentrated under re-
duced pressure, which gave 1.57 g of a solid, which was
recrystallized from ethanol and gave 1.4 g (93%) 1,4-bis(4-
(s, 6H), 2.04 (s, 6H), 2.03 (s, 6H), 2.01 (s, 6H), 1.60 (m, 4H),
1.35 (m, 4H), 1.24 (t, J ) 7.0 Hz, 4H) ppm. IR (NaCl): 3470,
1749 cm-1
.
St ep 6. 1,6-Bis(3-(3-hydroxyphenyl)-4-(2,3,4,6-tetra-O-
acetyl-R-D-mannnopyranosyloxyphenyl))hexane (342 mg, 0.307
mmol) was dissolved in THF (2 mL), and cesium carbonate
(0.2 g, 0.62 mmol) was added. The mixture was stirred for 10
min at room temperature, and then ethyl bromoacetate (0.07
mL, 0.63 mmol) was added. The mixture was stirred at room
temperature overnight, and then water was added. The
mixture was extracted with ethyl acetate, and the extracts
were combined, dried (MgSO4), and concentrated under re-
duced pressure, which gave 0.4 g (100%) of the ester, which
was used without further purification. 1H NMR (400 MHz,
CDCl3): 7.36 (t, J ) 7.7 Hz, 2H), 7.16 (m, 4H), 7.08 (d, J ) 1.2
Hz, 4H), 7.01 (m, 2H), 6.90 (m, 2H), 5.40 (s, 2H), 5.26 (m, 6H),
4.68 (d, J ) 2.6 Hz, 4H), 4.22 (m, 2H), 4.12 (m, 4H), 3.93 (J )
2.2, 12.0 Hz, 2H), 3.77 (m, 2H), 2.58 (t, J ) 7.2 Hz, 4H), 2.13
(s, 6H), 2.02 (s, 6H), 2.00 (s, 6H), 1.96 (s, 6H), 1.61 (m, 4H),
1
methoxybenzoyl)butane as a white solid, mp 137-138 °C. H
NMR (400 MHz, CDCl3): 7.93 (d, J ) 7.0 Hz, 4H), 6.92 (d, J
) 7.0 Hz, 4H), 3.86 (s, 6H), 2.97 (m, 4H), 1.82 (m, 4H) ppm.
IR (KBr): 1675 cm-1
.
The diketone (1.4 g, 4.29 mmol) in DMSO (10 mL) was
treated with hydrazine hydrate (0.3 mL, 9.44 mmol) and
warmed at 60 °C for 1 h, potassium tert-butoxide (1.44 g, 12.9
mmol) was added, and the reaction was heated at reflux. After
3 h, the reaction was cooled, mixed with water, and extracted
with ethyl acetate. The organic materials were combined,
dried (MgSO4), and concentrated under reduced pressure,
which gave 1.07 g (84%) of 40 as a clear oil which was used
without further purification. 1H NMR (400 MHz, CDCl3): 7.07
(d, J ) 7.0 Hz, 4H), 6.81 (d, J ) 7.0 Hz, 4H), 3.79 (s, 6H), 2.53
(m, 4H), 1.57 (m, 4H), 1.34 (m, 4H) ppm. IR (NaCl): 1612
1.37 (m, 4H), 1.30 (m, 6H) ppm. IR (KBr): 1752 cm-1
.
Step 7. The product from step 6 was dissolved in THF (5
mL) and then treated with 2 N sodium hydroxide (2.0 mL).
After 3 h, the mixture was acidified with 2 N HCl, and the
solvent was removed under reduced pressure. A portion was
purified by reverse-phase HPLC (10-100% acetonitrile in
water) which provided 56 mg of 42 as a white solid, mp 114-
118 °C. 1H NMR (400 MHz, DMSO-d6): 7.30 (t, J ) 7.7 Hz,
2H), 7.22 (d, J ) 8.8 Hz, 2H), 7.11 (m, 4H), 7.07 (d, J ) 7.7
Hz, 2H), 6.95 (m, 2H), 6.87 (m, 2H), 5.24 (s, 2H), 4.68 (s, 4H),
3.67 (m, 2H), 3.58 (d, J ) 10.2 Hz, 2H), 3.31-3.49 (m, 8H),
2.55 (t, J ) 7.0 Hz, 4H), 1.58 (m, 4H), 1.34 (m, 4H) ppm. IR
cm-1
.
Step 2. The bis-ether 40 (6.25 g, 20.9 mmol) was dissolved
in dichloromethane (100 mL) and cooled to 0 °C. Ferric
chloride (0.7 g, 1 mol %) was added, and then bromine (2.18
mL, 42.3 mmol) was added dropwise. The reaction was stirred
for 3 h and then mixed with water. The organic materials
were separated, dried (MgSO4), and then concentrated under
reduced pressure, which gave 9.27 g (98%) of a pale yellow oil
which solidified on standing, mp 84-85 °C. 1H NMR (400
MHz, CDCl3): 7.34 (d, J ) 1.8 Hz, 2H), 7.04 (dd, J ) 1.8, 8.0
Hz, 2H), 6.80 (d, J ) 8.0 Hz, 2H), 3.86 (s, 6H), 2.50 (t, J ) 8.0
Hz, 4H), 1.55 (m, 4H), 1.31 (m, 4H) ppm.
(KBr): 3414, 1735 cm-1
. MS (CI methane): m/e 571 [M -
(mannose)2]+. Anal. (C46H54O18‚0.8TFA) C, H.
Ack n ow led gm en t. One of the authors (B.D.) ac-
knowledges insightful discussions with Larry Denner
and J on O. Nagy. We thank Tony J . You and David S.
Maxwell for providing assistance with generating Figure
2, and Karin M. Keller for intellectual contributions to
methodology development and assistance with proof-
reading the manuscript. A sample of 4-(aminomethyl)-
1,8-octanediamine (triaminononane) was supplied by
Monsanto and is gratefully acknolwedged. We thank one
of the reviewers for bringing to our attention additional
relevant references regarding enhanced multivalent
binding.
Step 3. 1,6-Bis(3-bromo-4-methoxyphenyl)hexane (6.0 g,
13.15 mmol) was dissolved in dichloromethane (100 mL), and
cooled to 0 °C, and then treated dropwise with boron tribro-
mide (3.1 mL, 32.9 mmol). After 3 h, the reaction was
quenched with ice water, and the organic material was
separated, dried (MgSO4), and then concentrated under re-
duced pressure, which gave 5.0 g of an off-white solid which
was recrystallized from dichloromethane and gave 4.55 g (81%)
of a white solid, mp 63-65 °C. 1H NMR (400 MHz, CDCl3):
7.24 (d, J ) 1.8 Hz, 2H), 6.99 (dd, J ) 1.8, 8.0 Hz, 2H), 6.91
(d, J ) 8.0 Hz, 2H), 5.33 (s, 2H), 2.49 (t, J ) 7.7 Hz, 4H), 1.55
(m, 4H), 1.32 (m, 4H) ppm. IR (KBr): 3419 cm-1
.
Su p p or tin g In for m a tion Ava ila ble: Analytical data on
final compounds not contained in the Experimental Section
(4 pages). Ordering information is given on any current
masthead page.
Step 4. 1,6-Bis(3-bromo-4-hydroxyphenyl)hexane (1.0 g,
2.34 mmol) was dissolved in 1,2-dichloroethane (10 mL) and
glycosylated with R-D-mannose pentaacetate (2.3 g, 5.84 mmol)
as before, which gave 1.15 g (46%) of 41 as a clear oil. 1H
NMR (400 MHz, CDCl3): 7.35 (s, 2H), 7.01 (s, 4H), 5.61 (dd, J
) 3.3, 10.1 Hz, 2H), 5.51 (m, 4H), 5.38 (t, J ) 10.3 Hz, 2H),
4.27 (dd, J ) 5.1, 12.1 Hz, 2H), 4.17 (m, 2H), 4.06 (dd, J )
2.2, 12.1 Hz, 2H), 2.51 (t, J ) 8.0 Hz, 4H), 2.19 (s, 6H), 2.05
(s, 6H), 2.03 (s, 6H), 2.02 (s, 6H), 1.54 (m, 4H), 1.30 (m, 4H)
ppm.
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Step 5. The bis-glycoside 41 (446 mg, 0.41 mmol) was
dissolved in dimethoxyethane (3.0 mL) and treated with
3-hydroxyphenylboronic acid (124 mg, 0.9 mmol), bis(triphen-
ylphosphine)palladium(II) chloride (10 mg), and potassium
phosphate tribasic (348 mg, 1.64 mmol), and the mixture was
degassed and then heated at reflux for 2 h. The reaction was
mixed with 2N HCl (5 mL), then extracted with ethyl acetate.
The extracts were combined, dried (MgSO4), and concentrated
under reduced pressure. Purification by flash chromatography
(SiO2, gradient elution, hexane to ethyl acetate), gave 320 mg
(71%) of a clear oil. 1H NMR (400 MHz, CDCl3): 7.30 (t, J )
8.0 Hz, 2H), 7.25 (m, 4H), 7.07 (m, 4H), 7.01 (d, J ) 8.3 Hz,
2H), 6.88 (dd, J ) 2.6, 8.4 Hz, 2H), 6.64 (s, 2H), 5.51 (s, 2H),
5.42 (m, 2H), 5.40 (m, 2H), 5.33 (t, J ) 9.6 Hz, 2H), 4.18 (dd,
J ) 5.1, 12.4 Hz, 2H), 4.10 (q, J ) 7.0 Hz, 2H), 3.99 (dd, J )
2.2, 12.5 Hz, 2H), 3.88 (m, 2H), 2.57 (t, J ) 7.3 Hz, 4H), 2.15

Inhibitors of Selectin-Mediated Cell Adhesion
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Market, R. V.; Beck, P. J .; Voytus, J . A.; Revelle, B. M.; Scott,
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oligosaccharide Selectin Inhibitors: (R-D-Mannopyranosyloxy)-
biphenyl-Substituted Carboxylic Acids. J . Med. Chem. 1995, 38,
4976-4984. (b) Dupre´, B.; Bui, H.; Scott, I. L.; Market, R. V.;
Keller, K. M.; Beck, P. J .; Kogan, T. P. Glycomimetic Selectin
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(23) (a) Palma-Vargas, J . M.; Toledo-Pereyra, L. H.; Dean, R. E.;
Dixon, R. A. F.; Kogan, T. P. Small-Molecule Selectin Inhibitor
Protects Against Liver Inflammatory Response After Ischemia
and Reperfusion J . Am. Coll. Surgeons 1997, 185, 365-372. (b)
Abraham, W. M.; Ahmed, A.; Cortes, A.; Bjercke, R.; Hu, X.;
Revelle, B. M.; Kogan, T. P.; Dupre´ B., Scott I. L.; Dixon, R. A.
F.; Yeh, E. T. H.; Beck, P. J . Selectin Blockade Prevents Antigen-
Induced Late Broncial Responses and Airway Hyperresponsive-
ness in Allergic Sheep. Manuscript in preparation.
(9) (a) Sanders, W. J .; Katsumoto, T. R.; Bertozzi, C. R.; Rosen, S.
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evant Modifications of the Lewis x Trisaccharide. Biochemistry
1996, 35, 14862-14867. (b) Tsujishita, H.; Hiramatsu, Y.; Kondo,
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(10) Kogan, T. P.; Revelle, B. M.; Tapp, S.; Scott, D.; Beck, P. J . A
Single Amino Acid Residue can Determine the Ligand Specificity
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(24) Differences in IC50 values for sLex and 3 from those previously
reported8a are due to minor changes in the assay format. All
data presented in this table are from multiple assays, and the
values reported are consistent within this table.
J M9704917