Sulfated galactocerebrosides as potential antiinflammatory agents

Article abstract of DOI:10.1021/jm9606960

Native sulfatides, as well as many sulfated glycolipids, have been shown to avidly bind to the selectin receptors. In vivo, native sulfatides significantly block activity in selectin-dependent inflammatory responses. The fact that nonsulfated galactocerebrosides did not inhibit selectin- mediated adhesion identified a critical role for the anionic sulfate residue. We therefore initiated a program to evaluate the activity of position isomers. This study showed a binding selectivity for the positions 2 and 3 of the sulfate group on the carbohydrate ring as well as enhanced activity for the disulfated analogs. Furthermore, it was discovered that the attachment of lipophilic substituents on the carbohydrate ring was tolerated, consistent with the presence of a lipophilic pocket in the binding cavity. This resulted in compounds with a 6-fold increased potency.

Full text of DOI:10.1021/jm9606960

3234
J . Med. Chem. 1997, 40, 3234-3247
Su lfa ted Ga la ctocer ebr osid es a s P oten tia l An tiin fla m m a tor y Agen ts
Anne Marinier,*,‡ Alain Martel,‡ J acques Banville,‡ Carol Bachand,‡ Roger Remillard,‡ Philippe Lapointe,‡
Brigitte Turmel,‡ Marcel Menard,‡ William E. Harte, J r.,† J . J . Kim Wright,† Gordon Todderud,∇
Kenneth M. Tramposch,∇ J u¨rgen Bajorath,§ Diane Hollenbaugh,§ and Alejandro Aruffo§
Departments of Chemistry, Dermatological Disease, and Inflammatory Disease, Bristol-Myers Squibb Pharmaceutical Research
Institute, Candiac, Quebec, Canada, Wallingford, Connecticut, Buffalo, New York, and Seattle, Washington
Received October 7, 1996X
Native sulfatides, as well as many sulfated glycolipids, have been shown to avidly bind to the
selectin receptors. In vivo, native sulfatides significantly block activity in selectin-dependent
inflammatory responses. The fact that nonsulfated galactocerebrosides did not inhibit selectin-
mediated adhesion identified a critical role for the anionic sulfate residue. We therefore initiated
a program to evaluate the activity of position isomers. This study showed a binding selectivity
for the positions 2 and 3 of the sulfate group on the carbohydrate ring as well as enhanced
activity for the disulfated analogs. Furthermore, it was discovered that the attachment of
lipophilic substituents on the carbohydrate ring was tolerated, consistent with the presence of
a lipophilic pocket in the binding cavity. This resulted in compounds with a 6-fold increased
potency.
Ch a r t 1. Sulfatide Structurea
In tr od u ction
The adhesion of leukocytes to endothelial cells con-
stitutes an early and essential step in inflammatory cell
extravasation. One class of molecules that mediates
these interactions is the selectin family which includes
three known members: E-, P-, and L-selectins.1a These
receptors are type I membrane proteins and have a
homologous extracellular domain structure, including
an N-terminal lectin domain and an epidermal growth
factor-related domain, and a variable number of comple-
ment regulatory repeat elements. P-Selectin (CD-62),
whose expression is upregulated by thrombin and other
mediators, is found on the cell surface of platelets and
endothelial cells. E-Selectin is expressed by the vas-
cular endothelial cells after cytokine activation associ-
ated with inflammation. L-Selectin is expressed on
most leukocytes.1
Extravasation of leukocytes into sites of inflammation
is in part dependent on the initial interaction with
endothelial P-selectin. This is supported by the finding
of in vivo antiinflammatory activity of P-selectin-block-
ing antibodies.2 P-Selectin is thus a novel antiinflam-
matory target, and small molecular weight antagonists
may have useful antiinflammatory activity.
A terminal moiety composed of sialyl Lewis x (SLex)
on the surface of leukocytes was recognized as one
ligand involved in selectin-dependent cell adhesion.3 The
myeloid cell line HL-60, which expresses the SLex
antigen on its surface, was found to adhere strongly to
both P- and E-selectins. A second class of selectin
ligand, the sulfatides 1, was also identified.4 These
native sulfated galactocerebrosides were found to strongly
bind to P- and L-selectin IgG chimeras. In addition
sulfatides were shown to block the binding of HL-60
cells to P- and E-selectin Ig fusion proteins. Sulfatides
a
a , R ) (CH2)13CHdCH(CH2)7CH3 (nervonic); b, R ) CHOH-
(CH2)12CHdCH(CH2)7CH3 (R-hydroxynervonic); c, R ) (CH2)22CH3
(lignoceric); d , R ) (CHOH(CH2)21CH3 (R-hydroxylignoceric); e,
R ) (CH2)14CH3 (palmitic).
were recently shown to have antiinflammatory activity
in vivo.5 After intravenous administration sulfatides
caused a marked reduction in the vascular permeability
and cell infiltration in the rat dermal reverse passive
Arthus reaction. They also demonstrated significant
protective effects in neutrophil- and selectin-dependent
models of lung injury in rats. In addition, they were
recently reported as suppressors of the CCl4-induced
liver inflammation in rats by inhibiting the attachment
of L-selectin-expressing lymphocytes to their native
sugar ligands.
The sulfatides are a family of sulfated galactocerebro-
sides that differ only in the composition of the fatty acid
residue that acylates the amino group of the sphingosine
(Chart 1, 1). In sulfatides prepared from bovine brain,
the fatty acid component consists mainly of nervonic and
R-hydroxynervonic acids with a smaller percentage of
the corresponding saturated R-hydroxylated and non-
hydroxylated lignoceric acids.
Molecular models of E-selectin6 and P-selectin7 have
been constructed based on the X-ray structure of the
rat mannose-binding protein (MBP), a type C-lectin with
homology to the selectins in core regions and the lectin
domain. A shallow depression, close to the calcium ion
and solvent accessible, was found after examination of
the models. Site-specific mutagenesis of the residues
located in this groove showed the importance of the
region for the binding of HL-60 cells. In P-selectin,
critical residues within this groove include Lys 113, Tyr
48, and Tyr 94. These residues are conserved in E- and
L-selectins. The recently published crystal structure of
* To whom correspondence should be addressed: Bristol-Myers
Squibb Pharmaceutical Research Institute, 100 Boul. de l’Industrie,
Candiac, Quebec, Canada, J 5R 1J 1.
‡ Department of Chemistry, Quebec.
† Department of Chemistry, CT.
∇ Department of Dermatological Disease, NY.
§ Department of Inflammatory Disease, WA.
X Abstract published in Advance ACS Abstracts, August 15, 1997.
S0022-2623(96)00696-6 CCC: $14.00 © 1997 American Chemical Society

Sulfatides as Potential Antiinflammatory Agents
J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 20 3235
Sch em e 1a
a
(a) Hg(CN)2/CH3NO2/80-85 °C; (b) MeONa/MeOH/rt; (c) MeONa/MeOH/-50 °C.
the lectin and EGF domains of E-selectin has cor-
roborated these previous models.8
Mon osu lfa ted An a logs. Starting with the â-glyco-
side intermediate 5 or 6 standard carbohydrate protec-
tion-deprotection strategies were used for the prepa-
ration of the 2-, 3-, 4-, and 6-monosulfated analogs as
shown in Schemes 2 and 3. Hence, protection of
compound 6 with 2,2-dimethoxypropane gave the cor-
responding 3,4-O-isopropylidene derivative 7 which was
used to synthesize the 3- and 6-monosulfated analogs.
Selective monosulfation at the primary hydroxyl group
followed by removal of the 3,4-O-isopropylidene protect-
ing group produced azido triol 9 which was then reduced
and acylated as described by Schmidt et al.12 to give
compound 10. The 6-monosulfated compound 11 was
obtained after a final debenzoylation. On the other
hand, complete benzoylation at positions 2 and 6 of the
3,4-O-isopropylidene 7 followed by attachment of the
palmitoyl side chain on the reduced derivative afforded
the fully protected intermediate 13. This compound was
then hydrolyzed to give the corresponding 3,4-diol 14.
Introduction of an acetate in the axial position 4 was
accomplished as described by Lemieux,15 and the free
3-hydroxyl group of compound 15 was then sulfated.
Basic hydrolysis gave the 3-sulfated galactocerebroside
17.
The coupling of the palmitoic acid moiety could also
be achieved at the beginning of the synthetic sequence
as illustrated for the preparation of the 2-sulfated
analog 23 in Scheme 3. Thus, unprotected glycoside 6
was reduced and acylated, and the 3 and 4 positions of
the resulting galactocerebroside 18 were protected as
an isopropylene ketal. The primary hydroxyl group at
position 6 of diol 19 was then selectively blocked by
treatment with pivaloyl chloride. Sulfation of the
resulting alcohol 20 followed by protecting group cleav-
age gave the 2-sulfated galactocerebroside 23.
Mutagenesis studies using sulfatides as the binding
entity to P-selectin7b,9 revealed that sulfatides and the
cellular ligand on HL-60 cells bind to an overlapping
but not identical set of residues in P-selectin. Lys 113
remains critical for binding of P-selectin to both sul-
fatides and the cellular ligand. In contrast, Tyr 94 and
Tyr 48 are not crucial for binding to sulfatides, while
Lys 111, which was not critical for binding to myeloid
cells, is as important as Lys 113 for binding to sulfati-
des. In a model of sulfatides binding to P-selectin,9 the
negatively charged group in sulfatides should make an
ionic interaction with Lys 113 and/or Lys 111.
The relative chemical simplicity of the sulfatides
coupled to their antiinflammatory activity prompted us
to initiate a synthetic program aimed at further under-
standing of the key elements required for biological
activity and the augmentation of this activity. Careful
examination of the putative binding cavity revealed a
lipophilic pocket delimitated by the side chains of Arg
85 and Lys 84. Attachment of nonpolar substituents
at specific positions on the carbohydrate ring should
provide additional lipophilic contacts with the receptor
that could result in potency gain. Furthermore, the
presence of two positively charged residues in the
binding cavity (Lys 113 and Lys 111) encouraged the
preparation of polysulfated derivatives that could make
additional interactions with these residues. Here we
report some preliminary data on molecular modification
of sulfatides and the effect on the ability to bind to
P-selectin. Specifically we determine the optimum
position for sulfation and the effect of disulfation. We
also explore the incorporation of lipophilic substituents
on the galactose ring and their issue on the potency of
the compounds.
Finally, from the key intermediate 5, the 4 and 6
positions on the carbohydrate were protected by forma-
tion of the benzylidene acetal 24. Complete benzoyla-
tion followed by acidic treatment produced diol 26 where
the primary 6-hydroxyl group was subsequently blocked
as a pivaloyl ester. After reduction of the azido group
and acylation, alcohol 28 was sulfated and hydrolyzed
to afford the 4-sulfated galactocerebroside 30.
Disu lfa ted An a logs. The preparation of the disul-
fated analogs followed similar synthetic sequences as
described in Schemes 4 and 5. The 3,4-O-isopropy-
lidenegalactocerebroside 19 was first bis-sulfated and
then submitted to an acidic treatment to remove the
Ch em istr y
The pure â-anomer 4 was obtained from O-(2,3,4,6-
t et r a -O-a cet yl-R-D-ga la ct opyr a n osyl)t r ich lor oa cet -
imidate and 3 using Schmidt’s methodology.10,11 Alter-
natively, glycosidation of D-erythro-azidosphingosine12
(3) with tetra-O-acetyl-R-D-galactosyl bromide13 (2) un-
der Koenigs-Knorr conditions14 catalyzed with mercuric
cyanide was also performed and gave a mixture of â-
and R-anomeric glycoside 4 in 76% and 9.5% yields,
respectively (Scheme 1). The two anomers could be
conveniently separated at this stage.

3236 J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 20
Sch em e 2. 3- and 6-Monosulfated Compoundsa
Marinier et al.
a
PTSA/CH3C(OCH3)2CH3; (b) SO3‚N(CH3)3/DMF or Py‚SO3/pyridine; (c) TFA (90%); (d) H2S/pyridine/H2O; (e) CH3(CH2)14COCl/NaOAc/
THF; (f) NaOMe/MeOH; (g) BzCl/pyridine; (h) CH3C(OEt)3/PTSA/toluene; (i) HOAc/H2O.
isopropylidene ketal group. Debenzoylation gave the
2,6-disulfated glycoside 32.
a 96-well ELISA plate coated with sulfatides (Sigma).
The P-selectin was mixed with HRP-conjugated goat
anti-human IgG, and this immunocomplex was incu-
bated on the antigen-coated plate in the presence or
absence of the test compounds. The bound complexes
were detected by addition of buffer containing substrate
and 3,3′,5,5′-tetramethylbenzidine. Color development
was monitored, and following termination of the assay,
the absorbance at 450 nm was determined. The test
compounds were compared to sulfatides (positive con-
trol) or solvent (negative control). The compounds were
initially dissolved in DMSO and diluted in PBS so that
the final assay conditions contained 0.5% DMSO. Dose-
response curves were constructed for each compound
using eight concentrations in 2-fold serial dilutions. The
mean of duplicate determinations was used for each
data point. The IC50 values reported were interpolated
from the linear portion of the log dose-response curves.
The data are reported in Tables 1 and 2.
The synthesis of the 3,6-disulfated analog 41 started
with the 3,4-O-isopropylideneglycoside 33 which was
then selectively silylated at position 6. After benzoy-
lation of the free hydroxyl groups, the acylamino side
chain was introduced as before. Compound 36 was then
hydrolyzed, and the resulting diol 37 was submitted to
Lemieux’s sequence.15 This afforded, after treatment
with fluoride ions in the presence of acetic acid to avoid
acetate migration at position 6, the diol 39 which was
bis-sulfated. Basic hydrolysis provided the 3,6-sulfated
galactocerebroside 41.
The preparation of the 2,3- and 4,6-disulfated analogs
required the same intermediate 42 originating from
unprotected glycoside 6 (Scheme 5). The usual forma-
tion of the cerebroside side chain (reduction and acyla-
tion) was followed by sulfation of the free hydroxyl
groups in positions 2 and 3. Hydrolysis of the benzoyl
protecting group in compound 49 gave the 2,3-disulfated
galactocerebroside 50. On the other hand, bis-benzoy-
lation at positions 2 and 3 of the 4,6-O-benzylidene
acetal 42 and subsequent azido reduction and acylation
provided the fully protected derivative 43. The prepa-
ration of the 4,6-disulfated glycoside 46 was completed
by acidic cleavage of the benzylidene acetal, bis-sulfation
at positions 4 and 6 of the resulting diol 44, and final
debenzoylation.
Resu lts
Table 1 clearly shows that sulfatides strongly inhibit
the binding of P-selectin to sulfatide-coated plates. In
compound 17 the natural fatty acid side chain is
replaced by palmitic acid. This change had no effect
on the inhibition of binding. Therefore, all further
analogs were prepared as palmitates. The sulfate group
was absolutely required for inhibition since galactosyl
ceramide was completely inactive at 50 µM.
In the case of monosulfates the 3 position appears to
be optimum for inhibitory activity. The 3-sulfated
analog 17 was found to be 4-fold more potent than the
2-sulfated analog 23 (Table 1). The 4- and 6-monosul-
fated derivatives 30 and 11 are much less active with
20-fold decreased potencies compared to 17.
Biology
In Vitr o Cell-F r ee Bin d in g Assa y. The assay used
to determine the ability of compounds to inhibit the
binding of P-selectin has employed a truncated soluble
P-selectin protein containing the lectin, EGF, and two
complement repeat domains fused to an immunoglobu-
lin tail, termed receptor globulin (Rg), as previously
described.1,16 The P-selectin Rg was allowed to bind to
Sulfation of more than one galactose hydroxyl group
significantly enhances the ability to inhibit P-selectin

Sulfatides as Potential Antiinflammatory Agents
J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 20 3237
Sch em e 3. 2- and 4-Monosulfated Compoundsa
a
(a) H2S/pyridine/H2O; (b) CH3(CH2)14COCl/NaOAc/THF; (c) PTSA/CH3C(OCH3)2CH3; (d) C4H9COCl/pyridine; (e) Py‚SO3/pyridine or
NMe3‚SO3/DMF; (f) TFA (90%); (g) MeONa/MeOH; (h) PhCHO/HCO2H; (i) BzCl/pyridine; (j) TFA (50%)/CH2Cl2.
Ta ble 1. In Vitro Results of Various Sulfatide Derivatives as
Ta ble 2. In Vitro Results of Various Substituted Sulfatide
Inhibitors of P-Selectin
Derivatives as Inhibitors of P-Selectin
cell-free
cell-free
position(s)
of sulfate(s)
assay
IC50 (µM)
assay
IC50 (µM)
no.
no.
R1
Bz
R2
R3
R4
R6
commercial sulfatides
(Sigma)
11
17
23
30
32
41
3
0.64
10
16
31
40
45
48
49
H
H
H
SO3Na
Bz
SO3Na
SO3Na
SO3Na
0.4
4.3
0.9
1.3
0.1
0.1
0.1
0.64
Bz Bz
Bz SO3Na
Bz Bz
SO3Na Ac
6
3
2
4
2, 6
3, 6
4, 6
12
0.6
2.5
12.5
1.2
2.4
H
H
SO3Na Ac
Bz SO3Na
Bz Bz
Bz SO3Na SO3Na -CHPh- -CHPh-
Bz SO3Na SO3Na
SO3Na
H
H
H
H
sulfatides
(Sigma)
H
H
46
1.7
50
2, 3
0.1
galactosyl ceramide
no sulfate
>50
of replacing hydroxyl groups with lipophilic substitu-
ents. In compound 10, the hydroxyl group in the
aglycone of this 6-monosulfate was protected by a
benzoyl group. This change resulted in a 30-fold
improvement in activity relative to the parent com-
pound. When all of the hydroxyl groups were protected,
as in the 3-sulfated compound 16, there was a 7-fold
decrease in potency. This limited exploration of the
hydrophobic pocket suggested that lipophilic groups on
both the galactose and aglycone moieties could be
tolerated. The incorporation of a benzoyl function into
the aglycone was also tolerated in the disulfate analogs.
binding. The 2,6-disulfate (32) was found to be 10 times
more potent than the 6-monosulfate and 2 times more
potent than the 2-monosulfate. In general, all of the
disulfated analogs are more active than their monosul-
fated counterparts. The 2,3-disulfate compound 50 is
at least 10-fold more active than the other disulfates.
Moreover, 50 is 6-fold more potent than sulfatides.
We also evaluated a number of synthetic intermedi-
ates for inhibition of P-selectin binding (Table 2). We
were particularly interested in determining the effect

3238 J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 20
Sch em e 4. 2,6- and 3,6-Disulfated Compoundsa
Marinier et al.
a
(a) SO3‚N(CH3)3/DMF or SO3‚py/pyridine; (b) TFA (90%); (c) NaOCH3/CH3OH; (d) PTSA/CH3C(OCH3)2CH3; (e) TBDMSCI/pyridine;
(f) BzCl/pyridine; (g) H2S/pyridine/H2O; (h) CH3(CH2)14COCl/NaOAc/THF; (i) PTSA/CH3C(OEt)3/toluene; (j) HOAc, H2O; (k) TBAF/AcOH/
THF.
Sch em e 5. 2,3- and 4,6-Disulfated Compoundsa
a
(a) PhCHO/HCOOH; (b) BzCl/pyridine; (c) H2S/pyridine/H2O; (d) CH3(CH2)14COCl/NaOAc/THF; (e) TFA (90%)/CH2Cl2; (f) NMe3‚SO3/
DMF or Py‚SO3/pyridine; (g) NaOMe/MeOH.
The benzoylated 2,6-disulfate 31 and the 2,3-disulfate
49 had potencies equivalent to those of the parent
compounds 32 and 50. As in the monosulfate series,
derivatization of all remaining hydroxyl groups was
tolerated as exemplified by compounds 40 and 48. The
3,6-disulfated derivative 40 was 2-fold more potent than
the parent compound, and 48 was equiactive. In the
case of the polybenzoylated 4,6-disulfate 45, a 17-fold
improvement in inhibitory activity was observed over
the parent compound 46.

Sulfatides as Potential Antiinflammatory Agents
J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 20 3239
Discu ssion
Since this behavior cannot be ruled out from the more
active series (3- or 2,3-sulfated analogs), the contribution
of the benzoate group in ceramide aglycone in terms of
lipophilicity gain or protein interactions cannot yet be
assessed with certainty. It seems however that most
of the less active sulfated series (4,6- and 3,6-disulfated)
benefit from the presence of nonpolar substituents in
the carbohydrate ring as shown by compounds 45 and
40. Despite no significant activity improvement in
compound 48, the presence of a benzylidene group at
positions 4 and 6 of this 2,3-disulfated derivative
indicates also that lipophilic substituents are well-
tolerated. These results could be explained by the
presence of a lipophilic pocket in the binding cavity,
delimited by the carbon side chains of Lys 84 and Arg
85.18
This study shows that the position of the sulfate on
the galactose ring is important for optimal interaction
with P-selectin. The 3 position is most favored. The
rank order of potency was found to be 3 > 2 > 4 ) 6.
Surprisingly, despite the critical importance of the
sulfate residue for interaction with P-selectin (as ex-
emplified by the lack of activity of galactosyl ceramide)
all of the positional isomers retain binding affinity to
P-selectin. These results are similar to the findings
described by Suzuki and co-workers.4b They reported
the positional selectivity of the sulfate group in a series
of synthetic sulfatide analogs evaluated for the ability
to bind to L-selectin. Their rank order for the galactose
sulfate was 3 > 6 > 2. Therefore, the 3-sulfate appears
to be favored in the interaction of galactosyl-based
compounds with both P- and L-selectins. Our finding
that P-selectin prefers a 2-monosulfate over a 6-mono-
sulfate is not similar to Suzuki’s results. It is possible
that there may be subtle differences in the binding
pockets which can account for this apparent selectivity.
However, it must be noted that the assays used were
quite different. We evaluated the ability to inhibit
P-selectin binding to sulfatide-coated plates. The L-
selectin study evaluated the ability of the protein to bind
to plates coated with the test compound.
The finding that the sulfate group can be at different
positions on the pyranose ring and still retain significant
activity tends to indicate that the position of the
carbohydrate ring in the protein groove is not as critical
as we first thought. The ionic interaction of Lys 113
and/or Lys 111 with the sulfate entity can still be
accommodated to some extent in the different mono-
sulfates, despite a required rotation or translation of the
galactose ring. The high flexibility of the lipophilic
ceramide side chain can probably permit slight modi-
fications in its direction and tilt. This means that
different binding modes are likely tolerated, depending
on the position of the sulfate group.
Con clu sion
In this study we have shown that modifications of
sulfatides can produce compounds with enhanced bind-
ing affinity for P-selectin. Previous work using antibod-
ies in animal models demonstrated that the selectins
appear to play an important role in inflammatory cell
infiltration. In addition, sulfatides have shown to block
cell infiltration in rats, albeit high doses of compound
were required. Compounds with enhanced potency for
inhibiting P-selectin-dependent cell adhesion are desired
as potential therapeutic agents in the treatment of acute
inflammatory reactions.
The high flexibility of these analogs, nevertheless,
does not permit so far to unambiguously establish the
general binding features. Furthermore, the physical
properties of this family of compounds in terms of
formation of vesicles or micelles have to be more deeply
explored in order to elaborate a more precise molecular
binding model. Additional studies are underway to
further probe the sulfatide structural requirements for
P-selectin-dependent adhesion to provide insight on the
design of more potent and specific inhibitors.
The good potency of the disulfated analogs is consis-
tent with the importance of both Lys 111 and Lys 113
as described by Bajorath et al.7b,9 Therefore, it can be
presumed that the two sulfate groups can interact to a
certain extent with both positive residues. The impor-
tance of polysulfation was also observed by Suzuki4b in
that L-selectin binding was strongest to a 3,4,6-trisul-
fate. As we observed for the monosulfated analogs, the
position of these two negatively charged residues in the
galactose ring appears to be fairly specific. It has been
shown by Lemieux17 that specificity in the recognition
of oligosaccharides by proteins involves clusters of
hydroxyl groups in these oligosaccharides. These clus-
ters are usually essential to complex formation and have
been termed “polar key” by Lemieux. By analogy, the
current biological results tend to demonstrate that a
“polar key” located at positions 2 and 3 or 4 and 6 of
the galactose residue is preferred for binding activity,
as shown by compounds 48 and 45. Ionic groups to
nonadjacent positions like in compound 40 seem to be
less favored.
Exp er im en ta l Section
Ma ter ia ls a n d Meth od s. Analytical grade solvents were
used for reactions and chromatographies. Flash column
chromatographies were performed on Merck silica gel 60 (230-
400 Mesh), and Merck silica gel 60 F254 0.5 mm plates were
used. All melting points were determined on a Gallenkamp
melting point apparatus and are not corrected. 1H NMR
spectra were measured on a Bruker AC200 (200 MHz) or a
Bruker AMX400 (400 MHz) instrument. Chemical shifts are
reported in δ units using the solvent as internal standard. The
signals are described as s (singlet), d (doublet), t (triplet), qa
(quartet), qi (quintet), m (multiplet), and br (broad). Infrared
spectra were recorded on a Perkin-Elmer 781, and optical
rotations were measured on a Perkin-Elmer 241 apparatus.
Elemental analyses were performed at the Analytical Center
of Bristol-Myers Squibb in Syracuse, NY.
Key In ter m ediates. (2S,3R,4E)-2-Azido-3-(ben zoyloxy)-
1-[(2,3,4,6-tetr a -O-a cetyl-r-D-ga la ctop yr a n osyl)oxy]-4-oc-
tadecen e an d (2S,3R,4E)-2-Azido-3-(ben zoyloxy)-1-[(2,3,4,6-
tetr a-O-acetyl-â-D-galactopyr an osyl)oxy]-4-octadecen e (4).
P r oced u r e A: A solution of (2S,3R,4E)-2-azido-3-(benzoyloxy)-
4-octadecen-1-ol4 (3) (2.9 g, 6.75 mmol) in dry benzene (100
mL) and nitromethane (100 mL) was heated under reflux.
Benzene was distilled, and the solution was concentrated
under vacuum to 50-60 mL. To this solution were added
2,3,4,6-tetra-O-acetyl-R-D-galactosyl bromide5 (2) (5.0 g, 12.16
mmol) and mercury(II) cyanide (3.0 g, 12.16 mmol) at 22 °C
and under argon, and the resulting mixture was heated up to
80-85 °C for 15-20 min. The reaction was then cooled down
A benzoate group in the ceramide aglycone appears
to improve the activity of sulfated compounds at position
6 (cf. compounds 10 and 31). The same trend is noticed
for the 4,6- and 3,6-disulfated analogs 45 and 40,
although benzoate groups are also present in the sugar.

3240 J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 20
Marinier et al.
to 5 °C and diluted with ethyl ether/water (1:1, 100 mL).
Hydrogen sulfide was bubbled in, and the resulting black
precipitate was filtered on Celite and washed with ethyl ether
(3 × 25 mL). The organic phases were washed with cold
aqueous sodium bicarbonate solution (1 M, 4 × 25 mL), water
(25 mL), and brine (25 mL), dried over anhydrous magnesium
sulfate, filtered, and concentrated. The black resulting residue
was purified by chromatography on silica gel (250 g, 0-30%
ethyl acetate/hexane) and afforded the â-anomer (3.90 g, 76%)
and the R-anomer (0.49 g, 9.5%) of the title compound as
colorless oils.
P r oced u r e B (m od ifica tion of Sch m id t’s p r oced u r e):
(2S,3R,4E)-2-Azido-3-(benzoyloxy)-4-octadecen-1-ol (3.93 g, 9.14
mmol) and O-(2,3,4,6-tetra-O-acetyl-R-D-galactopyranosyl)-
trichloroacetimidate (6.30 g, 12.78 mmol) in a mixture of dry
dichloromethane (50 mL) and hexane (90 mL) were stirred
with crushed 4 Å molecular sieves (3 g) for 20 min. A solution
of tin(IV) chloride in dichloromethane (1.8 mL, 1 M) was then
added dropwise over 50 min. The mixture was stirred for
another 1 h after the addition ended. The sieves were filtered
and washed with ethyl acetate, and the filtrate was diluted
with ethyl acetate (300 mL). The solution was washed with
water, saturated sodium bicarbonate, and brine, dried over
anhydrous sodium sulfate, filtered, and concentrated. The
residue was purified by silica gel chromatography (toluene/
ethyl acetate) to afford the â-anomer (5.30 g, 76%). Only traces
of (2S,3R,4E)-2-azido-3-(benzoyloxy)-4-octadecen-1-yl acetate
were obtained using this method, while, in our hands, up to
40% of this side product was obtained using Schmidt’s original
procedure.
IR (CH2Cl2) νmax (cm-1) R-anomer: 3050, 2930 (C-H), 2100
(-N3), 1750 (CdO). IR (CH2Cl2) νmax (cm-1) â-anomer: 3050,
2930, 2955 (C-H), 2130 (N3), 1750 (CdO). 1H NMR (400 MHz,
CDCl3) δ R-anomer: 0.89 (3H, t, J ) 7.0 Hz, -CH3), 1.25 (20H,
br s, -(CH2)10-), 1.39 (2H, m, -CH2-), 2.00, 2.01, 2.09, 2.15 (4 ×
3H, 4s, 4 × -OCOCH3), 2.09 (2H, m, dCH-CH2-), 3.52 (1H, dd,
J ) 10.7, 7.7 Hz, H-1), 3.88 (1H, dd, J ) 10.7, 3.5 Hz, H-1),
3.91-3.95 (1H, m, H-2), 4.09-4.10 (2H, m, H-6′), 4.24 (1H,
td, J ) 6.5, 1.1 Hz, H-5′), 5.14-5.17 (2H, m, H-1′, H-2′), 5.34-
5.39 (1H, m, H-3′), 5.49 (1H, dd, J ) 3.3, 1.1 Hz, H-4′), 5.53-
5.60 (2H, m, H-3, H-4), 5.93-5.99 (1H, m, H-5), 7.45-8.06 (5H,
3m, -C6H5). 1H NMR (400 MHz, CDCl3) δ â-anomer: 0.89 (3H,
t, J ) 7.0 Hz, -CH3), 1.25 (20H, br s, -(CH2)10-), 1.39 (2H, m,
-CH2-), 2.00, 2.03, 2.11, 2.16 (4 × 3H, 4s, 4 × -OCOCH3), 2.09
(2H, m, dCH-CH2), 3.58-3.63 (1H, m, H-1), 3.89-3.97 (3H,
m, H-1, H-5′, H-2), 4.11 (1H, dd, J AB ) 11.2, J AX ) 6.7 Hz,
H-6′), 4.14 (1H, dd, J AB ) 11.2, J BX ) 6.7 Hz, H-6′), 4.51 (1H,
d, J ) 7.9 Hz, H-1′), 5.02 (1H, dd, J ) 10.5, 3.4 Hz, H-3′), 5.42
(1H, dd, J ) 10.5, 7.9 Hz, H-2′), 5.39 (1H, d, J ) 3.4 Hz, H-4′),
5.53-5.62 (2H, m, H-3, H-4), 5.94 (1H, dt, J ) 14.3, 6.9 Hz,
H-5), 7.45-8.08 (5H, 3m, -C6H5).
(2S,3R,4E)-2-Azid o-3-(ben zoyloxy)-1-(â-D-ga la ctop yr a -
n osyloxy)-4-octa d ecen e (6). A solution of (2S,3R,4E)-2-
azido-3-(benzoyloxy)-1-[(2,3,4,6-tetra-O-acetyl-â-D-galactopyr-
anosyl)oxy]-4-octadecene (4) (4.0 g, 5.26 mmol) in dichloro-
methane (20 mL) was added slowly to a freshly prepared
solution of sodium (2.44 g, 106 mmol) in methanol (120 mL)
at -70 °C and under argon. The temperature of the cooling
bath was allowed to reach -50 °C during 3 h. The reaction
mixture was cooled down to -70 °C and neutralized with a
solution of acetic acid (6.0 mL, 106 mmol) in dichloromethane
(10 mL). The mixture was concentrated under vacuum, giving
a residue which was dissolved in dichloromethane (50 mL).
The residual solid (sodium acetate) was filtered on Celite and
washed with dichloromethane (5 × 10 mL). The combined
filtrate and washings were evaporated, and the residue was
purified by silica gel chromatography (200 g, 0-12% methanol/
dichloromethane) and afforded the title compound (2.77 g,
89%) as a white solid.
H-1), 4.13 (1H, d, J ) 7.5 Hz, H-1′), 4.16-4.20 (1H, m,
-CHN3-), 4.91 (1H, br d, J ) 3.8 Hz, -OH), 4.55 (1H, ap t, -OH),
4.75 (1H, br s, -OH), 4.91 (1H, br d, J ) 3.6 Hz, -OH), 5.56
(1H, dd, J ) 15.3, 7.6 Hz, H-4), 5.65 (1H, dd, J ) 7.6, 3.7 Hz,
H-3), 5.88 (1H, dt, J ) 15.3, 6.8 Hz, H-5), 7.53-8.00 (5H, 3m,
-C6H5).
Mon osu lfa ted Com p ou n d s. Syn th esis of (2S,3R,4E)-
2-(Hexa d eca n oyla m in o)-3-h yd r oxy-1-[[6-O-(sod iu m oxy-
su lfon yl)-â-D-ga la ct op yr a n osyl]oxy]-4-oct a d ecen e (11).
(2S,3R,4E)-2-Azid o-3-(b en zoyloxy)-1-[(3,4-O-isop r op yl-
id en e-â-D-ga la ctop yr a n osyl)oxy]-4-octa d ecen e (7). A so-
lution of (2S,3R,4E)-2-azido-3-(benzoyloxy)-1-(â-D-galactopy-
ranosyloxy)-4-octadecene (6) (0.955 g, 0.161 mmol) in 2,2-
dimethoxypropane (50 mL) was treated with p-toluenesulfonic
acid (0.050 g), and the resulting mixture was stirred at 22 °C
for 18 h. Water (5 mL) was added followed by p-toluene-
sulfonic acid (0.045 g), and the mixture was stirred for another
1.5 h. Triethylamine (3 drops) was then added, and the
mixture was evaporated under vacuum. The residue was
dissolved in ethyl acetate (400 mL), and the organic phase was
washed with a saturated solution of sodium bicarbonate and
brine, dried over anhydrous sodium sulfate, and concentrated.
The residue was purified by silica gel chromatography (60 g,
20-30% acetone/toluene) and afforded the title compound
(0.600 g, 60%).
[R]22D: -17.3° (c ) 0.98, CHCl3). IR (NaCl, film) νmax (cm-1):
3650-3100 (O-H), 2930, 2855 (C-H), 2105 (-N3), 1720
(CdO ester). 1H NMR (200 MHz, CDCl3) δ: 0.87 (3H, t, J )
6.3 Hz, -CH3), 1.24-1.35 (22H, m, -(CH2)11-), 1.35 and 1.52 (2
× 3H, 2s, (CH3)2-C-), 2.03-2.13 (2H, m, dCH-CH2-), 2.33-
2.38 (1H, m, -OH-6′), 2.55 (1H, d, J ) 2.4 Hz, -OH-2′), 3.57
(1H, ddd, J ) 8.3, 8.3, 2.4 Hz, H-2′), 3.65-4.17 (8H, m, -CHN3-,
H-1, H-3′, H-4′, H-5′, H-6′), 4.22 (1H, d, J ) 8.3 Hz, H-1′), 5.58
(1H, dd, J ) 15.0, 8.0 Hz, H-4), 5.71 (1H, dd, J ) 8.0, 4.0 Hz,
H-3), 5.97 (1H, dt, J ) 15.0, 6.7 Hz, H-5), 7.39-8.08 (5H, 3m,
-C6H5).
(2S ,3R ,4E )-2-Azid o-3-(b e n zoyloxy)-1-[[3,4-O-isop r o-
p ylid en e-6-O-(sod iu m oxysu lfon yl)-â-D-ga la ct op yr a n o-
syl]oxy]-4-octa d ecen e (8). A solution of (2S,3R,4E)-2-azido-
3-(benzoyloxy)-1-[(3,4-O-isopropylidene-â-D-galactopyranosyl)-
oxy]-4-octadecene (7) (0.550 g, 0.87 mmol) in dry pyridine was
treated with sulfur trioxide-pyridine complex (0.207 g, 1.28
mmol), and the resulting mixture was stirred at 22 °C for 2.5
h. Water (5 mL) and solid sodium bicarbonate (0.50 g) were
added, and the mixture was stirred for 10 more minutes and
evaporated under vacuum. The residue was coevaporated with
toluene and purified by silica gel chromatography (chloroform/
methanol, 95:5-8:2) and afforded the title compound (0.532
g, 83%) as a white glassy solid.
[R]22D: 0° (c ) 0.8, MeOH, 95%). IR (Nujol) νmax (cm-1):
3700-3100 (br, O-H), 2900 (br, C-H), 2110 (-N3), 1715 (CdO
ester). 1H NMR (400 MHz, DMSO-d6) δ: 0.85 (3H, t, J ) 6.7
Hz, -CH3), 1.20-1.32 (22H, m, -(CH2)11-), 1.26 and 1.39 (2 ×
3H, 2s, -C(CH3)2), 2.04 (2H, qa, J ) 6.8 Hz, dCH-CH2-), 3.22
(1H, br qa, H-2′), 3.61 (1H, dd, J ) 10.3, 5.1 Hz, H-1), 3.80
(1H, dd, J ) 10.3, 8.0 Hz, H-1), 3.86 (2H, d, J ) 6.1 Hz, H-6′),
3.95 (1H, br t, H-3′), 4.00 (1H, br td, H-5′), 4.10 (1H, dd, J )
5.6, 1.8 Hz, H-4′), 4.17 (1H, m, H-2), 4.21 (1H, d, J ) 8.0 Hz,
H-1′), 5.30 (1H, d, J ) 5.3 Hz, -OH), 5.57 (1H, dd, J ) 15.1,
7.4 Hz, H-4), 5.64 (1H, dd, J ) 7.4, 3.6 Hz, H-3), 5.88 (1H, dt,
J ) 15.1, 6.8 Hz, H-5), 7.54-7.58, 7.65-7.69, and 7.97-7.99
(5H, 3m, aromatic H).
(2S,3R,4E)-2-Azido-3-(ben zoyloxy)-1-[[6-O-(sod iu m oxy-
su lfon yl)-â-D-ga la ctop yr a n osyl]oxy]-4-octa d ecen e (9). A
solution of (2S,3R,4E)-2-azido-3-(benzoyloxy)-1-[[3,4-O-isopro-
pylidene-6-O-(sodium oxysulfonyl)-â-D-galactopyranosyl]oxy]-
4-octadecene (8) (0.42 g, 0.57 mmol) in methanol (60 mL) was
treated with Dowex H+ resin (1 g) for 45 min. The mixture
was filtered and the resin washed with chloroform/methanol
(7:3, 20 mL). The filtrate was treated with Rexyn Na+ resin
(∼2 g) until the solution became basic. The mixture was
filtered and evaporated, and the residue was purified by silica
gel chromatography (20-30% methanol/chloroform) and af-
forded the title compound (0.358 g, 90%) as a waxy solid.
IR (Nujol) νmax (cm-1): 3700-3100 (br, O-H), 2915, 2850 (C-
H), 2110 (-N3), 1720 (CdO ester). 1H NMR (400 MHz, DMSO-
d6) δ: 0.84 (3H, t, J ) 6.8 Hz, -CH3), 1.19-1.31 (22H, m,
IR (CH2Cl2) νmax (cm-1): 3700-3200 (O-H), 3060, 2930,
2860 (C-H), 2110 (-N3), 1720 (CdO). 1H NMR (400 MHz,
DMSO-d6) δ: 0.85 (3H, t, J ) 6.8 Hz, -CH3), 1.20-1.65 (22H,
m, -(CH2)11-), 2.04 (2H, m, dCH-CH2-), 3.23-3.33 (3H, m, H-5′,
H-3′, H-2′), 3.44 (1H, dd, J ) 10.6, 5.7 Hz, H-6′), 3.51 (1H, dd,
J ) 10.6, 6.1 Hz, H-6′), 3.59 (1H, dd, J ) 10.5, 5.3 Hz, H-1),
3.61 (1H, d, J ) 2.1 Hz, H-4′), 3.78 (1H, dd, J ) 10.5, 7.5 Hz,

Sulfatides as Potential Antiinflammatory Agents
J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 20 3241
-(CH2)11-), 2.03 (2H, br qa, dCH-CH2-), 3.26-3.30, 3.54-3.60,
and 3.74-3.79 (2H, 3H, 2H, 3 sets of m, H-2′, H-3′, H-4′, H-5′,
H-6′, H-1), 3.83 (1H, dd, J ) 10.6, 6.2 Hz, H-1 or H-6′), 4.13
(1H, d, J ) 7.0 Hz, H-1′), 4.12-4.17 (1H, m, H-2), 4.50 (1H, d,
J ) 4.7 Hz, -OH), 4.66 (1H, d, J ) 5.3 Hz, -OH), 4.84 (1H, d,
J ) 4.5 Hz, -OH), 5.55 (1H, dd, J ) 15.3, 7.5 Hz, H-4), 5.63
(1H, dd, J ) 7.5, 3.6 Hz, H-3), 5.87 (1H, dt, J ) 15.3, 7.2 Hz,
H-5), 7.53-7.56, 7.64-7.68, and 7.96-7.98 (5H, 3 sets of m,
aromatic H).
in pyridine (15 mL) was treated with benzoyl chloride (0.66
mL, 5.68 mmol) at 5 °C. The mixture was stirred overnight
at 5 °C; then methanol (∼2 mL) was added to this cold mixture
which was stirred for 1 more hour. The mixture was diluted
with ethyl acetate (60 mL) and washed with 1 M aqueous
sodium bicarbonate (2 × 15 mL), water (2 × 15 mL), and brine
(15 mL). The organic phase was dried over anhydrous
magnesium sulfate, filtered, and concentrated. The residue
was purified by silica gel chromatography (0-16% ethyl
acetate/hexane) to give the title material (1.14 g, 96%) as a
yellow oil.
(2S,3R,4E)-2-(H exa d eca n oyla m in o)-3-(b en zoyloxy)-1-
[[6-O-(sod iu m oxysu lfon yl)-â-D-ga la ctop yr a n osyl]oxy]-4-
octa d ecen e (10). A solution of (2S,3R,4E)-2-azido-3-(ben-
zoyloxy)-1-[[6-O-(sodium oxysulfonyl)-â-D-galactopyranosyl]oxy]-
4-octadecene (9) (0.034 g, 0.049 mmol) in a mixture of pyridine
(2.5 mL) and water (2.5 mL) was saturated with hydrogen
sulfide and stirred at 22 °C for 18 h. The solvent was
evaporated under vacuum, and the last traces of pyridine were
coevaporated with toluene. The residual solid was then
dissolved in tetrahydrofuran (6 mL) and treated with aqueous
sodium acetate (50%, 5 mL) and palmitoyl chloride (0.016 g,
0.058 mmol). The aqueous phase was extracted with tetrahy-
drofuran (2 × 10 mL). The combined organic layers were
evaporated in vacuum, and the solid residue was purified by
silica gel chromatography (20-30% methanol/chloroform) to
give the title material (0.037 g, 84%) as a white glassy solid.
IR (CH2Cl2) νmax (cm-1): 3050, 2990, 2930, 2855 (C-H), 2110
(-N3), 1725 (CdO ester). 1H NMR (400 MHz, CDCl3) δ: 0.88
(3H, m, -CH3), 1.20-1.30 (22H, m, -(CH2)11-), 1.37 and 1.64 (2
× 3H, 2s, -C(CH3)2), 1.90 (2H, m, dCH-CH2-), 3.57 and 3.84-
3.98 (1H, 2H, 2 sets of m, H-1, H-2), 4.21 (2H, br t, H-5′), 4.32
(1H, dd, J ) 5.5, 1.9 Hz, H-4′), 4.41 (1H, br t, H-3′), 4.58 (1H,
d, J ) 7.9 Hz, H-1′), 4.56-4.73 (2H, m, H-6′), 5.28 (1H, br t,
H-2′), 5.44 (1H, dd, J ) 14.9, 8.0 Hz, H-4), 5.55 (1H, dd, J )
8.0, 3.3 Hz, H-3), 5.71 (1H, dt, J ) 14.9, 6.6 Hz, H-5), 7.37-
7.57 and 7.98-8.09 (15H, 2 sets of m, aromatic H).
(2S,3R,4E)-2-(H exa d eca n oyla m in o)-3-(b en zoyloxy)-1-
[(3,4-O-isop r op ylid en e-2,6-d i-O-ben zoyl-â-D-ga la ctop yr a -
n osyl)oxy]-4-octa d ecen e (13). (2S,3R,4E)-2-Azido-3-(ben-
zoyloxy)-1-[(3,4-O-isopropylidene-2,6-di-O-benzoyl-â-D-galac-
topyranosyl)oxy]-4-octadecene (12) (1.0 g, 1.19 mmol) was
reacted by the procedure used to synthesize compound 10 and
afforded the title compound (1.16 g, 93%) as a white solid. IR
(CH2Cl2) νmax (cm-1): 3680 (N-H), 3050, 2990, 2930, 2855 (C-
H), 1725, 1670 (CdO ester, amide). 1H NMR (400 MHz,
CDCl3) δ: 0.89 (6H, 2 t, J ) 6.7, 6.8 Hz, 2 × -CH3), 1.12-1.35
(46H, m, -(CH2)13-, -(CH2)10-), 1.38 and 1.64 (2 × 3H, 2s,
-C(CH3)2), 1.38-1.47 (2H, m, -CH2-), 1.80 (2H, t, J ) 7.6 Hz,
-NHCOCH2-), 1.97 (2H, qa, J ) 6.9 Hz, dCH-CH2-), 3.62 (1H,
dd, J ) 10.0, 4.0 Hz, H-1), 4.13 (1H, dd, J ) 10.0, 3.2 Hz, H-1),
4.17 (1H, m, H-5′), 4.31 (1H, dd, J ) 5.5, 2.1 Hz, H-4′), 4.41
(1H, dd, J ) 7.0, 5.5 Hz, H-3′), 4.39-4.45 (1H, m, H-2), 4.48
(1H, dd, J ) 11.6, 7.4 Hz, H-6′), 4.51 (1H, d, J ) 7.9 Hz, H-1′),
4.63 (1H, dd, J ) 11.6, 4.9 Hz, H-6′), 5.24 (1H, br t, H-2′), 5.45
(1H, dd, J ) 15.2, 7.2 Hz, H-4), 5.53 (1H, br t, H-3), 5.75 (1H,
d, J ) 9.1 Hz, -NH-), 5.80 (1H, dt, J ) 15.2, 6.9 Hz, H-5), 7.39-
7.46, 7.50-7.60, and 7.99-8.05 (15H, 3 sets of m, aromatic
H).
[R]22
: +4° (c ) 0.5, CHCl3/MeOH, 7:3). IR (KBr) νmax
D
(cm-1): 3700-3150 (br, O-H, N-H), 2930, 2860, (C-H), 1725,
1640 (CdO ester, amide). 1H NMR (400 MHz, DMSO-d6) δ:
0.85 (6H, t, J ) 6.8 Hz, 2 × -CH3), 1.21-1.35 (46H, m, -(CH2)10-,
-(CH2)13-), 1.47 (2H, m, -CH2-), 1.97-2.13 (4H, m, dCH-CH2-,
-NHCOCH2-), 3.28-3.31 (2H, m, H-2′, H-3′), 3.48 (1H, dd, J
) 10.2, 5.0 Hz, H-1), 3.54 (1H, br t, H-5′), 3.59 (1H, br t, H-4′),
3.74 (1H, dd, J AB ) 10.6 Hz, J AX ) 6.3 Hz, H-6′), 3.81 (1H, dd,
J AB ) 10.6 Hz, J BX ) 5.9 Hz, H-6′), 3.89 (1H, dd, J ) 10.2, 5.8
Hz, H-1), 4.07 (1H, d, J ) 7.1 Hz, H-1′), 4.34 (1H, m, H-2),
4.49 (1H, d, J ) 4.7 Hz, -OH), 4.66 (1H, br s, -OH), 4.92 (1H,
d, J ) 2.9 Hz, -OH), 5.45 (1H, dd, J ) 7.6, 5.8 Hz, H-3), 5.52
(1H, dd, J ) 15.1, 7.7 Hz, H-4), 5.79 (1H, dt, J ) 15.1, 6.8 Hz,
H-5), 7.48-7.52, 7.61-7.65, 7.93-7.95 (5H, 3 sets of m,
aromatic H), 7.84 (1H, d, J ) 9.1 Hz, -NH-).
(2S,3R,4E)-2-(Hexadecan oylam in o)-1-[[6-O-(sodiu m oxy-
su lfon yl)-â-D-ga la ctop yr a n osyl]oxy]-4-octa d ecen e (11). A
solution of (2S,3R,4E)-2-(hexadecanoylamino)-3-(benzoyloxy)-
1-[[6-O-(sodium oxysulfonyl)-â-D-galactopyranosyl]oxy]-4-oc-
tadecene (10) (0.402 g, 0.444 mmol) in methanol (25 mL) and
dichloromethane (25 mL) was treated with sodium methoxide
(0.2 M, 3.0 mL, 0.6 mmol) at 22 °C. The mixture was stirred
for 18 h. Resin Dowex H+ 50W-X8 was then added, and this
mixture was stirred until the pH became neutral. The mixture
was filtered, and the resin was washed with dichloromethane/
methanol, 1:1. The filtrate was then treated with resin Rexyn
102 (Na+) and stirred for 15 min. The mixture was filtered,
the resin was washed with dichloromethane/methanol (1:1),
and the filtrate was concentrated. The residue was purified
by silica gel chromatography (20% methanol/chloroform) and
afforded the title compound (0.257 g, 72%) as a white solid.
[R]22D: -6.5° (c ) 1.0, CHCl3/MeOH, 7:3). IR (KBr) νmax
(cm-1): 3700-3050 (br, O-H, N-H), 2920, 2850 (C-H), 1630
(CdO amide). 1H NMR (400 MHz, DMSO-d6) δ: 0.85 (6H, t,
J ) 6.8 Hz, 2 × -CH3), 1.23 (46H, m, -(CH2)13-, -(CH2)10-), 1.44
(2H, m, -CH2-), 1.92 (2H, m, dCH-CH2-), 2.02 (2H, t, J ) 7.4
Hz, -NHCOCH2-), 3.28 (2H, m, H-2′, H-3′), 3.38 (1H, dd, J )
9.9, 3.5 Hz, H-1), 3.54 (1H, t, J ) 6.1 Hz, H-5′), 3.59 (1H, br s,
H-4′), 3.75-3.88 (4H, m, H-2, H-3, H-6′), 3.97 (1H, dd, J )
9.9, 4.8 Hz, H-1), 4.03 (1H, d, J ) 7.0 Hz, H-1′), 4.49 (1H, d, J
) 4.7 Hz, -OH), 4.66 (1H, d, J ) 5.2 Hz, -OH), 4.87 (1H, d, J
) 3.3 Hz, -OH), 4.89 (1H, d, J ) 5.6 Hz, -OH), 5.34 (1H, dd, J
) 15.3, 7.0 Hz, H-4), 5.52 (1H, dt, J ) 15.3, 6.6 Hz, H-5), 7.52
(1H, d, J ) 9.0 Hz, -NH-).
(2S,3R,4E)-2-(H exa d eca n oyla m in o)-3-(b en zoyloxy)-1-
[(2,6-d i-O-b e n zoyl-â-D-ga la ct op yr a n osyl)oxy]-4-oct a -
d ecen e (14). A solution of (2S,3R,4E)-2-(hexadecanoylamino)-
3-(benzoyloxy)-1-[(3,4-O-isopropylidene-2,6-di-O-benzoyl-â-D-
galactopyranosyl)oxy]-4-octadecene (13) (1.15 g, 1.09 mmol) in
dichloromethane (40 mL) was treated with aqueous trifluoro-
acetic acid (90%, 1 mL) at 5 °C. The mixture was stirred at
22 °C for 1 h, then diluted with ethyl acetate (100 mL), and
washed with 1 M aqueous sodium bicarbonate (2 × 20 mL),
water (3 × 20 mL), and brine (20 mL). The organic phase was
dried over anhydrous magnesium sulfate, filtered, and evapo-
rated to give the title compound (1.06 g, 95% crude).
IR (CH2Cl2) νmax (cm-1): 3700-3450 (N-H, O-H), 3050,
2990, 2930, 2860 (C-H), 1725, 1670 (CdO ester, amide). 1H
NMR (400 MHz, CDCl3 + D2O) δ: 0.89 (6H, m, 2 × -CH3),
1.15-1.40 (48H, m, -(CH2)13-, -(CH2)11-), 1.83 (2H, t, J ) 7.6
Hz, -NHCOCH2-), 1.98 (2H, qa, J ) 6.9 Hz, dCH-CH2-), 3.62
(1H, dd, J ) 9.4, 3.2 Hz, H-3′), 3.80-3.83 (2H, m, H-1, H-5′),
3.98 (1H, d, J ) 3.2 Hz, H-4′), 4.13 (1H, d, J ) 9.9 Hz, H-1),
4.21 (1H, dd, J ) 11.4, 6.5 Hz, H-6′), 4.44 (1H, m, H-2), 4.52-
4.56 (1H, m overlapped by H-1′, H-6′), 4.53 (1H, d, J ) 7.7
Hz, H-1′), 5.26 (1H, br t, H-2′), 5.46 (1H, dd, J ) 15.2, 7.9 Hz,
H-4), 5.59 (1H, br t, H-3), 5.81 (1H, d, J ) 9.0 Hz, -NH-), 5.85
(1H, dt, J ) 15.2, 6.9 Hz, H-5), 7.39-7.47, 7.51-7.61, and
8.01-8.05 (15H, 3 sets of m, aromatic H).
(2S,3R,4E)-2-(H exa d eca n oyla m in o)-3-(b en zoyloxy)-1-
[(2,6-d i-O-ben zoyl-4-O-a cetyl-â-D-ga la ctop yr a n osyl)oxy]-
4-octa d ecen e (15). A suspension of (2S,3R,4E)-2-(hexade-
canoylamino)-3-(benzoyloxy)-1-[(2,6-di-O-benzoyl-â-D-galac-
topyranosyl)oxy]-4-octadecene (14) (1.06 g, 1.05 mmol) in dry
benzene (25 mL) and triethyl orthoacetate (25 mL) was treated
with p-toluenesulfonic acid (25 mg) at 22 °C. The mixture was
stirred for 1 h, and then triethylamine (∼1 mL) was added
Syn th esis of (2S,3R,4E)-2-(Hexa d eca n oyla m in o)-3-h y-
d r oxy-1-[[3-O-(sod iu m oxysu lfon yl)-â-D-ga la ctop yr a n o-
syl]oxy]-4-octa d ecen e (17). (2S,3R,4E)-2-Azid o-3-(ben -
zoyloxy)-1-[(3,4-O-isop r op ylid en e-2,6-d i-O-b en zoyl-â-D-
ga la ctop yr a n osyl)oxy]-4-octa d ecen e (12). A solution of
(2S,3R,4E)-2-azido-3-(benzoyloxy)-1-[(3,4-O-isopropylidene-â-
D-galactopyranosyl)oxy]-4-octadecene (7) (0.900 g, 1.42 mmol)

3242 J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 20
Marinier et al.
followed by cold water (20 mL). The aqueous phase was
extracted with ethyl acetate (4 × 50 mL), and these combined
extracts were washed with 1 M aqueous sodium bicarbonate
(3 × 40 mL), water (2 × 40 mL), and brine (40 mL). The
organic phase was dried over anhydrous magnesium sulfate,
filtered, and concentrated. The residue was dissolved in
dichloromethane (∼5 mL) and treated with 80% aqueous acetic
acid (10 mL). The mixture was stirred at 22 °C for ∼ 15 min.
Toluene was added, and the mixture was coevaporated under
vacuum (3x). The residue was purified by silica gel chroma-
tography (0-40% ethyl acetate/hexane) and afforded the title
compound (1.1 g, 100%) as a white solid.
Syn th esis of (2S,3R,4E)-2-(Hexa d eca n oyla m in o)-3-h y-
d r oxy-1-[[2-(sod iu m oxysu lfon yl)-â-D-ga la ctop yr a n osyl]-
oxy]-4-octa d ecen e (23). (2S,3R,4E)-2-(Hexa d eca n oyla m i-
n o)-3-(b en zoyloxy)-1-(â-D-ga la ct op yr a n osyloxy)-4-oct a -
d ecen e (18). (2S,3R,4E)-2-Azido-3-(benzoyloxy)-1-(â-D-galac-
topyranosyloxy)-4-octadecene (6) (250 mg, 0.42 mmol) was
reacted by the procedure used to synthesize compound 10 and
afforded the title compound (260 mg, 77%) as a white solid.
IR (CH2Cl2) νmax (cm-1): 3700-3100 (O-H, N-H), 3050,
2930, 2860 (C-H), 1720 (CdO ester), 1670 (CdO amide). 1H
NMR (400 MHz, CDCl3) δ: 0.87-0.90 (6H, ≈t, 2 × -CH3),
1.24-1.63 (48H, br m, -(CH2)13-, -(CH2)11-), 2.04 (2H, m, dCH-
CH2-), 2.17-2.22 (2H, m, -NHCOCH2-), 3.43 (1H, br t, H-5′),
3.53 (1H, dd, J ) 9.5, 3.3 Hz, H-3′), 3.66 (1H, dd, J ) 9.5, 7.7
Hz, H-2′), 3.71 (1H, dd, J ) 12.3, 3.9 Hz, H-1), 3.78-3.85 (2H,
m, H-6′), 3.99-4.03 (2H, m, H-4′, H-1), 4.29 (1H, d, J ) 7.7
Hz, H-1′), 4.57-4.60 (1H, m, H-2), 5.50 (1H, dd, J ) 15.3, 7.4
Hz, H-4), 5.65 (1H, t, J ) 7.4 Hz, H-3), 5.92 (1H, dt, J ) 15.3,
6.9 Hz, H-5), 6.16 (1H, d, J ) 6.16 Hz, -NH-), 7.45-8.04 (5H,
3 sets of m, -C6H5).
(2S,3R,4E)-2-(H exa d eca n oyla m in o)-3-(b en zoyloxy)-1-
[(3,4-O-isop r op ylid en e-â-D-ga la ctop yr a n osyl)oxy]-4-octa -
d ecen e (19). (2S,3R,4E)-2-(Hexadecanoylamino)-3-(benzoy-
loxy)-1-(â-D-galactopyranosyloxy)-4-octadecene (18) (0.170 g,
0.22 mmol) was reacted by the procedure used to synthesize
compound 7 and afforded the title compound (0.160 g, 90%)
as a white amorphous solid.
IR (CH2Cl2) νmax (cm-1): 3680, 3600, 3425 (O-H, N-H),
3050, 2930, 2850 (C-H), 1720 (CdO ester), 1640 (CdO amide).
1H NMR (400 MHz, CDCl3) δ: 0.89 (6H, t, J ) 6.7 Hz, 2 ×
-CH3), 1.25-1.35 (46H, m, -(CH2)13-, -(CH2)10-), 1.35 and 1.51
(2 × 3H, 2s, -C(CH3)2-), 1.57-1.66 (2H, m, -CH2-), 2.02-2.08
and 2.18-2.21 (2 × 2H, 2 sets of m, -NHCOCH2-, dCH-CH2-),
3.51 (1H, dd, J ) 7.1, 8.3 Hz, H-2′), 3.76-3.86 (3H, m, H-1,
H-6′, H-5′), 3.95 (1H, dd, J ) 11.5, 7.1 Hz, H-1), 3.98 (1H, dd,
J ) 11.2, 5.5 Hz, H-6′), 4.08 (1H, dd, J ) 7.1, 5.6 Hz, H-3′),
4.13 (1H, dd, J ) 5.6, 2.1 Hz, H-4′), 4.21 (1H, d, J ) 8.3 Hz,
H-1′), 4.55 (1H, m, H-2), 5.52 (1H, ddt, J ) 15.4, 7.2, 1.3 Hz,
H-4), 5.62 (1H, br t, H-3), 5.90 (1H, dt, J ) 15.4, 6.6 Hz, H-5),
5.97 (1H, d, J ) 9.2 Hz, -NH-), 7.44-7.53, 7.57-7.61, and
8.03-8.05 (5H, 3 sets of m, aromatic H).
IR (CH2Cl2) νmax (cm-1): 3700-3450 (N-H, O-H), 3050,
2930, 2860 (C-H), 1725, 1670 (CdO ester, amide). 1H NMR
(400 MHz, CDCl3) δ: 0.89 (6H, m, 2 × -CH3), 1.11-1.41 (48H,
m, -(CH2)13-, -(CH2)11-), 1.81 (2H, br t, -NHCOCH2-), 1.97 (2H,
qa, J ) 6.9 Hz, dCH-CH2-), 2.25 (3H, s, -OAc), 2.55 (1H, br s,
-OH), 3.66 (1H, dd, J ) 9.9, 3.8 Hz, H-1), 3.99 (1H, t, J ) 6.6
Hz, H-5′), 4.05 (1H, dd, J ) 10.0, 3.6 Hz, H-3′), 4.15 (1H, dd,
J ) 9.9, 3.2 Hz, H-1), 4.25 (1H, dd, J AB ) 11.3 Hz, J AX ) 6.5
Hz, H-6′), 4.30 (1H, dd, J AB ) 11.3 Hz, J BX ) 6.7 Hz, H-6′),
4.45 (1H, m, H-2), 4.50 (1H, d, J ) 7.9 Hz, H-1′), 5.27 (1H, dd,
J ) 10.0, 7.9 Hz, H-2′), 5.45 (1H, dd, J ) 15.2, 7.5 Hz, H-4),
5.47 (1H, br d, H-4′), 5.52 (1H, br t, H-3), 5.68 (1H, d, J ) 9.2
Hz, -NH-), 5.82 (1H, dt, J ) 15.2, 6.9 Hz, H-5), 7.39-7.47,
7.52-7.61, and 7.97-8.05 (15H, 3 sets of m, aromatic H).
(2S,3R,4E)-2-(H exa d eca n oyla m in o)-3-(b en zoyloxy)-1-
[[2,6-d i-O-ben zoyl-4-O-a cetyl-3-O-(sod iu m oxysu lfon yl)-
â-D-ga la ctop yr a n osyl]oxy]-4-octa d ecen e (16). A solution
of (2S,3R,4E)-2-(hexadecanoylamino)-3-(benzoyloxy)-1-[(2,6-di-
O-benzoyl-4-O-acetyl-â-D-galactopyranosyl)oxy]-4-octadecene
(15) (1.1 g, 1.04 mmol) in dry dimethylformamide (40 mL) was
treated with sulfur trioxide-trimethylamine complex (0.723 g,
5.20 mmol) and heated to 80-85 °C for 1.5 h. The mixture
was cooled down to 5 °C, and 1 M aqueous sodium bicarbonate
(15 mL) and solid sodium bicarbonate were added to satura-
tion. The mixture was stirred for 1 h and then concentrated
under vacuum. The residue was in dichloromethane, and this
mixture was filtered. The filtrate was evaporated and gave a
residue which was purified by silica gel chromatography (0-
12% methanol/chloroform) to afford the title material (1.09 g,
90%).
(2S,3R,4E)-2-(H exa d eca n oyla m in o)-3-(b en zoyloxy)-1-
[(3,4-O-isop r op ylid en e-6-O-p iva loyl-â-D-ga la ct op yr a n o-
syl)oxy]-4-octa d ecen e (20). A solution of (2S,3R,4E)-2-
(hexadecanoylamino)-3-(benzoyloxy)-1-[(3,4-O-isopropylidene-
â-D-galactopyranosyl)oxy]-4-octadecene (19) (80 mg, 0.1 mmol)
in pyridine (2 mL) was treated with pivaloyl chloride (0.015
mL, 0.12 mmol) and 4-(dimethylamino)pyridine (∼5-10 mg).
The mixture was stirred at 22 °C overnight, then diluted with
ethyl acetate (10 mL), and washed with water (3 × 5 mL), 1
M aqueous sodium bicarbonate (3 × 5 mL), water (3 × 5 mL),
and brine (5 mL). The organic phase was dried over anhydrous
magnesium sulfate, filtered, and evaporated. The residue was
purified by silica gel plates (50% ethyl acetate/hexane) and
afforded the title compound (64 mg, 73%) as a white amor-
phous solid.
IR (Nujol) νmax (cm-1): 3700-3350 (N-H), 2930, 2860 (C-
H), 1725, 1660 (CdO ester, amide). 1H NMR (400 MHz,
DMSO-d6) δ: 0.83 (6H, t, J ) 6.7 Hz, 2 × -CH3), 1.10-1.39
(48H, m, -(CH2)13-, -(CH2)11-), 1.74-1.96 (4H, m, dCH-CH2-,
-NHCOCH2-), 2.09 (3H, s -OAc), 3.52 (1H, dd, J ) 9.9, 7.1 Hz,
H-1), 3.72 (1H, dd, J ) 9.9, 6.6 Hz, H-1), 4.16 (1H, dd, J )
11.0, 7.1 Hz, H-6′), 4.24 (1H, br t, H-5′), 4.27-4.35 (2H, m,
H-2, H-6′), 4.65 (1H, dd, J ) 10.3, 3.4 Hz, H-3′), 4.83 (1H, d,
J ) 8.0 Hz, H-1′), 5.15 (1H, dd, J ) 10.3, 8.0 Hz, H-2′), 5.31
(1H, dd, J ) 7.4, 4.7 Hz, H-3), 5.38 (1H, dd, J ) 15.0, 7.3 Hz,
H-4), 5.46 (1H, dt, J ) 15.0, 6.3 Hz, H-5), 5.59 (1H, d, J ) 3.3
Hz, H-4′), 7.41-7.51, 7.56-7.65, 7.84-7.86, and 7.93-7.98
(16H, 4 sets of m, aromatic H, -NH-). MS (CI, NH3): 1132 (M
- Na)-.
(2S,3R,4E)-2-(Hexa d eca n oyla m in o)-3-h yd r oxy-1-[[3-O-
(sod iu m oxysu lfon yl)-â-D-ga la ctop yr a n osyl]oxy]-4-octa -
d ecen e (17). (2S,3R,4E)-2-(Hexadecanoylamino)-3-(benzoy-
loxy)-1-[[2,6-di-O-benzoyl-4-O-acetyl-3-O-(sodium oxysulfonyl)-
â-D-galactopyranosyl]oxy]-4-octadecene (16) (0.689 g, 0.59
mmol) was reacted by the procedure used to synthesize
compound 11 and afforded the title compound (0.316 g, 67%)
as a white solid.
IR (CH2Cl2) νmax (cm-1): 3690, 3500, 3430 (O-H, N-H),
3050, 2930, 2850 (C-H), 1725 (CdO ester), 1665 (CdO amide).
1H NMR (400 MHz, CDCl3) δ: 0.89 (6H, t, J ) 6.8 Hz, 2 ×
-CH3), 1.18 (9H, s, -OCOtBu), 1.25-1.37 (46H, m, -(CH2)13-,
-(CH2)10-), 1.33 and 1.49 (2 × 3H, 2s, -C(CH3)2-), 1.57-1.63 (2H,
m, -CH2-), 2.04 (2H, qa, J ) 7.0 Hz, dCH-CH2-), 2.17-2.19
(2H, m, -NHCOCH2-), 3.2 (1H, br s, -OH), 3.48 (1H, dd, J )
7.0, 7.2 Hz, H-2′), 3.76 (1H, dd, J ) 11.1, 3.8 Hz, H-1), 3.96
(1H, ddd, J ) 7.3, 5.8, 2.2 Hz, H-5′), 4.02 (1H, dd, J ) 11.1,
6.8 Hz, H-1), 4.07 (1H, dd, J ) 6.9, 5.5 Hz, H-3′), 4.11 (1H, dd,
J ) 5.5, 2.1 Hz, H-4′), 4.18 (1H, d, J ) 8.2 Hz, H-1′), 4.26 (1H,
IR (Nujol) νmax (cm-1): 3700-3050 (N-H), 2930, 2860 (C-
H), 1740, 1650 (CdO ester, amide). 1H NMR (400 MHz,
DMSO-d6) δ: 0.82 (6H, t, J ) 6.8 Hz, 2 × -CH3), 1.20-1.25
(46H, m, -(CH2)13-, -(CH2)11-), 1.40 (2H, m, -CH2-), 1.89 (2H,
m, dCH-CH2-), 1.99 (2H, t, J ) 7.4 Hz, -NHCOCH2-), 3.33-
3.56 and 3.91-3.95 (5H, 3H, 2 sets of m, H-1, H-2′, H-3′, H-4′,
H-3, H-6′), 3.73 (1H, m, H-2), 3.85 (1H, t, J ) 7.8 Hz, H-5′),
4.13 (1H, d, J ) 7.7 Hz, H-1′), 4.42 (1H, d, J ) 4.7 Hz, -OH),
4.58 (1H, t, J ) 5.5 Hz, -OH-6′), 4.84 (1H, d, J ) 5.4 Hz, -OH),
5.07 (1H, d, J ) 2.4 Hz, -OH), 5.30 (1H, dd, J ) 15.4, 7.3 Hz,
H-4), 5.50 (1H, dt, J ) 15.4, 6.7 Hz, H-5), 7.55 (1H, d, J ) 9.0
Hz, -NH-).
dd, J AB ) 11.4 Hz, J AX ) 7.3 Hz, H-6′), 4.31 (1H, dd, J AB
)
11.4 Hz, J BX ) 5.7 Hz, H-6′), 4.52 (1H, m, H-2), 5.49 (1H, ddt,
J ) 15.0, 7.1, 1.3 Hz, H-4), 5.56 (1H, dd, J ) 7.1, 5.5 Hz, H-3),
3.88 (1H, dt, J ) 15.0, 6.9 Hz, H-5), 6.01 (1H, d, J ) 9.1 Hz,
-NH-), 7.44-7.48, 7.53-7.61, and 8.02-8.04 (5H, 3 sets of m,
aromatic H).
(2S,3R,4E)-2-(H exa d eca n oyla m in o)-3-(b en zoyloxy)-1-
[[3,4-O-isop r op ylid en e-6-O-p iva loyl-2-O-(sod iu m oxysu l-
fon yl)-â-D-ga la ct op yr a n osyl]oxy]-4-oct a d ecen e
(21).
(2S,3R,4E)-2-(Hexadecanoylamino)-3-(benzoyloxy)-1-[(3,4-O-

Sulfatides as Potential Antiinflammatory Agents
J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 20 3243
isopropylidene-6-O-pivaloyl-â-D-galactopyranosyl)oxy]-4-octa-
decene (20) (61 mg, 0.066 mmol) was reacted by the procedure
used to synthesize compound 16 and afforded the title com-
pound (47 mg, 69%) as a white solid.
br s, -OH), 2.53 (1H, d, J ) 8.9 Hz, -OH-3′), 2.69 (1H, br s,
-OH), 3.51 (1H, d, J ) 1.0 Hz, H-5′), 3.57 (1H, m, H-2), 3.70
(1H, ddd, J ) 9.4, 8.9, 3.7 Hz, H-3′), 3.80 (1H, dd overlapped
by H-1, J ) 9.2, 8.1 Hz, H-2′), 3.81 (1H, dd, J ) 10.7, 3.9 Hz,
H-1), 4.09 (1H, dd, J ) 12.5, 1.8 Hz, H-6′), 4.16 (1H, dd, J )
10.6, 5.5 Hz, H-1), 4.22 (1H, dd, J ) 3.6, 0.8 Hz, H-4′), 4.30
(1H, br t, H-3), 4.34 (1H, d, J ) 12.5, 1.4 Hz, H-6′), 4.37 (1H,
d, J ) 7.6 Hz, H-1′), 5.53 (1H, ddt, J ) 15.4, 7.3, 1.2 Hz, H-4),
5.56 (1H, s, -OCHO-), 5.83 (1H, dt, J ) 15.4, 6.8 Hz, H-5),
7.36-7.40 and 7.49-7.52 (5H, 2 sets of m, aromatic H).
(2S,3R,4E)-2-Azid o-3-(ben zoyloxy)-1-[(2,3-d i-O-ben zoyl-
4,6-O-b e n zylid e n e -â-D-ga la ct op yr a n osyl)oxy]-4-oct a -
d ecen e (25). (2S,3R,4E)-2-Azido-1-[(4,6-O-benzylidene-â-D-
galactopyranosyl)oxy]-4-octadecene (24) (0.597 g, 1.03 mmol)
was reacted by the procedure used to synthesize compound
12 and gave the title compound (1.21 g, crude, quantitive)
which was used as such for the next reaction without any
further purification.
IR (CH2Cl2) νmax (cm-1): 3680 (N-H), 3050, 2930, 2850 (C-
H), 1725 (CdO ester), 1670 (CdO amide). 1H NMR (400 MHz,
CDCl3) δ: 0.89 (6H, br t, 2 × -CH3), 1.17 (9H, s, -OCOtBu),
1.17-1.29 (48H, m, -(CH2)13-, -(CH2)11-), 1.29 and 1.54 (2 ×
3H, 2s, -C(CH3)2-), 1.96 (2H, br qa, dCH-CH2-), 2.22 (2H, m,
-NHCOCH2-), 3.6-4.6 (10H, br dd, s, br t, 6 sets of m, H-2,
H-1, H-1′, H-2′, H-3′, H-4′, H-5′, H-6′), 5.45 (1H, dd, J ) 15.3,
7.1 Hz, H-4), 5.60 (1H, t, J ) 7.1 Hz, H-3), 5.88 (1H, dt, J )
15.3, 6.4 Hz, H-5), 6.66 (1H, br s, -NH-), 7.34-7.43, 7.53-7.56,
8.00-8.02 (5H, 3 sets of m, aromatic H).
(2S,3R,4E)-2-(H exa d eca n oyla m in o)-3-(b en zoyloxy)-1-
[[6-O-p iva loyl-2-O-(sod iu m oxysu lfon yl)-â-D-ga la ctop y-
r a n osyl]oxy]-4-oct a d ecen e (22). (2S,3R,4E)-2-(Hexade-
canoylamino)-3-(benzoyloxy)-1-[[3,4-O-isopropylidene-6-O-
pivaloyl-2-O-(sodium oxysulfonyl)-â-D-galactopyranosyl]oxy]-
4-octadecene (21) (52 mg, 0.05 mmol) was reacted by the
procedure used to synthesize compound 14 and afforded the
title compound (∼50 mg, crude) which was used for the next
step without purification.
IR (CH2Cl2) νmax (cm-1): 3050, 2930, 2860 (C-H), 2110
(-N3), 1730 (CdO), 1265 (C-O). 1H NMR (400 MHz, CDCl3)
δ: 0.89 (3H, t, J ) 6.8 Hz, -CH3), 1.23-1.33 (22H, m, -(CH2)11-
), 1.93 (2H, m, dCH-CH2-), 3.69-3.74 (2H, m, H-1, -CHN3-),
3.97-4.02 (2H, m, H-1, H-5′), 4.15 (1H, dd, J ) 12.4, 1.5 Hz,
H-6′), 4.42 (1H, dd, J ) 12.4, 1.3 Hz, H-6′), 4.61 (1H, d, J )
3.4 Hz, H-4′), 4.83 (1H, d, J ) 8.0 Hz, H-1′), 5.40 (1H, dd, J )
10.4, 3.4 Hz, H-3′), 5.46-5.56 (2H, m overlapping -O-CH-O-,
H-4, H-3), 5.56 (1H, s, -O-CH-O-), 5.73 (1H, dt, J ) 15.2, 6.7
Hz, H-5), 5.91 (1H, dd, J ) 10.4, 8.0 Hz, H-2′), 7.27-8.04 (20
H, 2m, 4 × -C6H5).
IR (CH2Cl2) νmax (cm-1): 3680-3200 (N-H, O-H), 3050,
2930, 2860 (C-H), 1725 (CdO ester), 1650 (CdO amide). 1H
NMR (200 MHz, CDCl3) δ: 0.9 (6H, br t, 2 × -CH3), 1.0 (9H,
s, -OCOtBu), 1.2 (46H, m, -(CH2)13-, -(CH2)10-), 1.5 (2H, m,
-CH2-), 1.9 and 2.2 (2 × 2H, 2 sets of m, -NHCOCH2-, dCH-
CH2-), 3.4-4.6 (10H, 5 sets of m, H-2, H-1, H-1′, H-2′, H-3′,
H-4′, H-5′, H-6′), 5.4-6.0 (4H, 3 sets of m, H-5, H-4, H-3,
-NH-), 7.4-7.6 and 8.01-8.04 (5H, 3 sets of m, aromatic H).
(2S,3R,4E)-2-(Hexa d eca n oyla m in o)-3-h yd r oxy-1-[[2-O-
(sod iu m oxysu lfon yl)-â-D-ga la ctop yr a n osyl]oxy]-4-octa -
d ecen e (23). (2S,3R,4E)-2-(Hexadecanoylamino)-3-(benzoy-
loxy)-1-[[6-O-pivaloyl-2-O-(sodium oxysulfonyl)-â-D-galactopyran-
osyl]oxy]-4-octadecene (22) (50 mg, 0.05 mmol) was reacted
by the procedure used to synthesize compound 11 and gave
the title compound (30 mg, 75%) as a beige solid.
(2S,3R,4E)-2-Azid o-3-(ben zoyloxy)-1-[(2,3-d i-O-ben zoyl-
â-D-ga la ctop yr a n osyl)oxy]-4-octa d ecen e (26). A solution
of (2S,3R,4E)-2-azido-3-(benzoyloxy)-1-[(2,3-di-O-benzoyl-4,6-
O-benzylidene-â-D-galactopyranosyl)oxy]-4-octadecene (25) (1.21
g, 1.03 mmol, crude) in dichloromethane (50 mL) was treated
with 50% aqueous trifluoroacetic acid (10 mL) at 22 °C. The
resulting mixture was stirred for 3 h, then cooled down to 5
°C, and neutralized with 1 M aqueous sodium bicarbonate and
solid sodium bicarbonate. The aqueous phase was extracted
with ethyl acetate (3 × 100 mL), and the combined extracts
were washed with 1 M aqueous sodium bicarbonate (2 × 25
mL), water (2 × 25 mL), and brine (25 mL). The organic phase
was dried over anhydrous magnesium sulfate, filtered, and
concentrated. The residue was purified by silica gel chroma-
tography (0-50% ethyl acetate/hexane) and afforded the title
compound (0.729 g, 88%) as a yellow oil.
IR (Nujol) νmax (cm-1): 3700-3100 (N-H, O-H), 2920, 2850
(C-H), 1635 (CdO amide). 1H NMR (400 MHz, DMSO-d6) δ:
0.85 (6H, t, J ) 6.8 Hz, 2 × -CH3), 1.23-1.31 (46H, m, -(CH2)13-,
-(CH2)10-), 1.41 (2H, m, -CH2-), 1.90 (2H, m, dCH-CH2-), 2.08
(2H, m, -NHCOCH2-), 3.25-3.39 and 3.46-3.53 (2H, 3H, 2 sets
of m, H-4′ or H-5′, H-6′, H-1), 3.66 (1H, br t, H-4′ or H-5′),
3.71 (1H, dt, J ) 9.0, 3.0 Hz, H-2), 3.86 (1H, br qa, H-3), 4.11
(1H, dd, J ) 9.5, 7.6 Hz, H-2′), 4.13 (1H, dd, J ) 9.3, 2.8 Hz,
H-3′), 4.23 (1H, d, J ) 7.6 Hz, H-1′), 4.56 (1H, d, J ) 3.8 Hz,
-OH), 4.61 (1H, t, J ) 5.6 Hz, -OH-6′), 4.81 (1H, d, J ) 5.6 Hz,
-OH), 4.99 (1H, d, J ) 1.4 Hz, -OH-3′), 5.31 (1H, dd, J ) 15.3,
7.2 Hz, H-4), 5.49 (1H, dt, J ) 15.3, 6.6 Hz, H-5), 7.39 (1H, d,
J ) 9.2 Hz, -NH-). MS (CI, NH3): 778 (M - Na)-.
Syn th esis of (2S,3R,4E)-2-(Hexa d eca n oyla m in o)-3-h y-
d r oxy-1-[[4-O-(sod iu m oxysu lfon yl)-â-D-ga la ctop yr a n o-
syl]oxy]-4-octa d ecen e (30). (2S,3R,4E)-2-Azid o-1-[(4,6-O-
ben zyliden e-â-D-galactopyr an osyl)oxy]-4-octadecen e (24).
A solution of (2S,3R,4E)-2-azido-3-(benzoyloxy)-1-[(2,3,4,6-
tetra-O-acetyl-â-D-galactopyranosyl)oxy]-4-octadecene (4) (1.0
g, 1.3 mmol) was treated as described by Schmidt13 to give
(2S,3R,4E)-2-azido-1-(â-D-galactopyranosyloxy)-4-octadecene (5).
This crude material was then dissolved in formic acid (5 mL)
and treated with benzaldehyde (5 mL), and this mixture was
stirred at 22 °C for 0.75 h. The mixture was then diluted with
ethyl acetate (50 mL), cooled down to 5 °C, and neutralized
with solid sodium bicarbonate and 1 M aqueous sodium
bicarbonate. The organic phases were separated, and the
aqueous phase was extracted with ethyl acetate (2 × 30 mL).
The combined organic extracts were washed with 1 M aqueous
sodium bicarbonate (30 mL), water (30 mL), and brine (30 mL).
The organic phase was dried over anhydrous magnesium
sulfate, filtered, and concentrated. The residue was purified
by silica gel chromatography (0-90% ethyl acetate/hexane)
and afforded the title compound (0.418 g, 56%) as a white solid.
IR (CH2Cl2) νmax (cm-1): 3600-3400 (O-H), 3050, 2930,
2860 (C-H), 2100 (-N3). 1H NMR (400 MHz, CDCl3) δ: 0.89
(3H, t, J ) 6.8 Hz, -CH3), 1.27-1.35 (20H, m, -(CH2)10-), 1.40
(2H, m, -CH2-), 2.08 (2H, qa, J ) 6.8 Hz, dCH-CH2-), 2.49 (1H,
IR (CH2Cl2) νmax (cm-1): 3700-3400 (O-H), 3050, 2930,
2860 (C-H), 2110 (-N3), 1730 (CdO). 1H NMR (400 MHz,
CDCl3) δ: 0.89 (3H, t, J ) 6.9 Hz, -CH3), 1.23-1.34 (22H, m,
-(CH2)11-), 2.01 (2H, qa, J ) 6.9 Hz, dCH-CH2-), 3.71-3.75
and 4.00-4.06 (2 × 2H, 2 sets of m, H-1, H-2, H-5′, H-6′), 3.87
(1H, dd, J ) 10.9, 6.2 Hz, H-1), 3.95 (1H, dd, J ) 12.3, 3.8 Hz,
H-6′), 4.45 (1H, br d, H-4′), 4.78 (1H, d, J ) 7.9 Hz, H-1′), 5.29
(1H, dd, J ) 10.3, 3.2 Hz, H-3′), 5.54 (1H, ddt, J ) 15.5, 8.1,
1.3 Hz, H-4), 5.76 (1H, dd, J ) 8.1, 4.2 Hz, H-3), 5.85 (1H, dd,
J ) 10.3, 7.9 Hz, H-2′), 5.88 (1H, dt, J ) 15.5, 6.9 Hz, H-5),
7.35-7.39, 7.44-7.53, 7.56-7.61, 7.95-8.01, and 8.04-8.10
(15H, 5 sets of m, aromatic H).
(2S,3R,4E)-2-Azid o-3-(ben zoyloxy)-1-[(2,3-d i-O-ben zoyl-
6-O-pivaloyl-â-D-galactopyr an osyl)oxy]-4-octadecen e (27).
(2S,3R,4E)-2-Azido-3-(benzoyloxy)-1-[(2,3-di-O-benzoyl-â-D-ga-
lactopyranosyl)oxy]-4-octadecene (26) (0.711 g, 0.89 mmol) was
reacted by the procedure used to synthesize compound 20 and
afforded the title compound (0.254 g, 32%) as a colorless oil
along with (2S,3R,4E)-2-azido-3-(benzoyloxy)-1-[(2,3-di-O-ben-
zoyl-4,6-di-O-pivaloyl-â-D-galactopyranosyl)oxy]-4-octadec-
ene (0.460 g, 53%) as a yellow oil.
IR (CH2Cl2) νmax (cm-1): 3700-3400 (O-H), 3050, 2930,
2860 (C-H), 2110 (-N3), 1730 (CdO). 1H NMR (400 MHz,
CDCl3) δ: 0.89 (3H, t, J ) 6.9 Hz, -CH3), 1.17-1.32 (31H, m,
-(CH2)11-, -OCOtBu), 1.95 (2H, qa, J ) 6.7 Hz, dCH-CH2-), 2.42
(1H, br s, -OH), 3.64 and 3.91-3.97 (1H, 3H, qa, m, H-1, H-2,
H-5′), 4.24 (1H, br s, H-4′), 4.32 (1H, dd, J AB ) 11.4, J AX ) 6.5
Hz, H-6′), 4.40 (1H, dd, J AB ) 11.4, J BX ) 6.4 Hz, H-6′), 4.74
(1H, d, J ) 7.9 Hz, H-1′), 5.33 (1H, dd, J ) 10.3, 3.3 Hz, H-3′),
5.48 (1H, dd, J ) 15.3, 8.0 Hz, H-4), 5.57 (1H, dd, J ) 8.1, 3.6
Hz, H-3), 5.72-5.79 (1H, m overlapped by H-2′, H-5), 5.77 (1H,

3244 J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 20
Marinier et al.
dd, J ) 10.3, 7.9 Hz, H-2′), 7.35-7.59 and 7.96-8.04 (15H, 2
sets of m, aromatic H).
(2H, m, dCH-CH2-), 2.02-2.21 (2H, m, -NHCOCH2-), 3.45-
3.51 (2H, m, H-1, H-3′), 3.58 (1H, br t, J ) 6.1 Hz, H-5′), 3.62
(1H, t, J ) 3.9 Hz, H-4′), 3.74 (1H, dd, J ) 10.6, 6.3 Hz, H-6′),
3.82 (1H, dd, J ) 10.6, 5.7 Hz, H-6′), 3.97 (1H, dd, J ) 9.8, 4.5
Hz, H-1), 4.15 (1H, dd, J ) 9.4, 7.8 Hz, H-2′), 4.24-4.28 (1H,
m, H-2), 4.27 (1H, d, J ) 7.6 Hz, H-1′), 4.69 (1H, d, J ) 3.9
Hz, -OH), 5.09 (1H, d, J ) 1.4 Hz, -OH), 5.39 (1H, t, J ) 7.6
Hz, H-3), 5.45 (1H, dd, J ) 15.0, 7.6 Hz, H-4), 5.76 (1H, dt, J
) 15.0, 6.8 Hz, H-5), 7.73 (1H, d, J ) 8.9 Hz, -NH-), 7.48-
7.98 (5H, 3m, -C6H5).
(2S,3R,4E)-2-(Hexa d eca n oyla m in o)-3-h yd r oxy-1-[[2,6-
bis-O-(sod iu m oxysu lfon yl)-â-D-ga la ctop yr a n osyl]oxy]-4-
octa d ecen e (32). (2S,3R,4E)-3-(Benzoyloxy)-2-(hexadecanoy-
lamino)-1-[[2,6-bis-O-(sodium oxysulfonyl)-â-D-galactopyran-
osyl]oxy]-4-octadecene (31) (0.243 g, 0.24 mmol) was reacted
by the procedure used to synthesize compound 11 and afforded
the title compound (0.17 g, 78%) as a pale beige solid.
IR (Nujol) νmax (cm-1): 3700-3100 (O-H, N-H), 2920, 2860
(C-H), 1650 (CdO amide). 1H NMR (400 MHz, DMSO-d6) δ:
0.85 (6H, t, J ) 6.8 Hz, 2 × -CH3), 1.23-1.43 (48H, br m,
-(CH2)13-, -(CH2)11-), 1.89 (2H, m, dCH-CH2-), 2.00-2.16 (2H,
m, -NHCOCH2-), 3.27 (1H, dd, J ) 9.3, 3.3 Hz, H-1), 3.47 (1H,
dd, J ) 9.6, 3.1 Hz, H-3′), 3.58 (1H, br t, H-5′), 3.63 (1H, d, J
) 3.1 Hz, H-4′), 3.70 (1H, m, H-3), 3.76-3.86 (1H, m overlap-
ping H-6′, H-2), 3.79 (1H, dd, J ) 10.6, 6.5 Hz, H-6′), 3.84 (1H,
dd, J ) 10.6, 5.6 Hz, H-6′), 4.09 (1H, dd, J ) 9.6, 7.7 Hz, H-2′),
4.15 (1H, dd, J ) 9.3, 2.7 Hz, H-1), 4.24 (1H, d, J ) 7.7 Hz,
H-1′), 4.68 (1H, br s, -OH), 4.87 (1H, d, J ) 3.8 Hz, -OH), 4.95
(1H, br s, -OH), 5.30 (1H, dd, J ) 15.4, 7.2 Hz, H-4), 5.48 (1H,
dt, J ) 15.4, 6.6 Hz, H-5), 7.41 (1H, d, J ) 9.2 Hz, -NH-).
Syn t h esis of (2S,3R,4E)-3-H yd r oxy-2-(h exa d eca n oyl-
a m in o)-1-[[3,6-bis-O-(sod iu m oxysu lfon yl)-â-D-ga la ctop y-
r a n osyl]oxy]-4-oct a d ecen e (41). (2S,3R,4E)-2-Azid o-3-
h yd r oxy-1-[(3,4-O-isop r op ylid en e-â-D-ga la ctop yr a n osyl)-
oxy]-4-octa d ecen e (33). (2S,3R,4E)-2-Azido-3-hydroxy-1-(â-
D-galactopyranosyloxy)-4-octadecene (5) (0.50 g, 1.07 mmol)
was reacted by the procedure used to synthesize compound 7
and afforded the title compound (0.425 g, 75%).
1H NMR (200 MHz, CDCl3) δ: 0.88 (3H, t, J ) 6.8 Hz, -CH3),
1.25 (22H, m, -(CH2)11-), 1.35 and 1.53 (2 × 3H, 2s, (CH3)2-C-),
1.64-2.13 (5H, m, dCH-CH2-, 3 × -OH), 3.44 (1H, m, H-2),
3.59 (1H, t, J ) 7.4 Hz, H-1), 3.81-4.35 (9H, m, H-1′, H-2′,
H-3′, H-4′, H-5′, H-6′, H-1, H-3), 5.53 (1H, m, H-4), 5.86 (1H,
m, H-5).
(2S,3R,4E)-2-Azido-3-h ydr oxy-1-[[3,4-O-isopr opyliden e-
6-O-(ter t-bu tyld im eth ylsilyl)-â-D-ga la ctop yr a n osyl]oxy]-
4-octa d ecen e (34). tert-Butyldimethylsilyl chloride (425 mg,
2.82 mmol) was added to a stirred solution of (2S,3R,4E)-2-
azido-3-hydroxy-1-[(3,4-O-isopropylidene-â-D-galactopyran-
osyl)oxy]-4-octadecene (33) (425 mg, 0.805 mmol) in pyridine
(20 mL) at -20 °C and under argon. The reaction mixture
was stirred at -20 °C overnight; then methanol was added.
The mixture was stirred again for 2 more hours at 22 °C. The
reaction mixture was then poured into water and diluted with
ethyl acetate (150 mL). The organic layer was washed with
water (5×) and brine, dried over anhydrous magnesium
sulfate, filtered, and concentrated. The residue was purified
by silica gel chromatography and afforded the title compound
(537 mg, 100%).
1H NMR (200 MHz, CDCl3) δ: 0.08 (6H, s, -Si(CH3)2), 0.88
(3H, t, J ) 6.7 Hz, -CH2-CH3), 0.90 (9H, s, -C(CH3)3), 1.25 (22H,
m, -(CH2)11-), 1.34 and 1.53 (2 × 3H, 2s, (CH3)2-C-), 1.66-2.11
(4H, m, dCH-CH2-, 2 × -OH), 3.45 (1H, m, H-2), 3.57 (1H, t,
H-1), 3.80-4.32 (9H, m, H-1′, H-2′, H-3′, H-4′, H-5′, H-6′, H-1,
H-3), 5.53 (1H, m, H-4), 5.84 (1H, m, H-5).
(2S,3R,4E)-2-Azid o-3-(ben zoyloxy)-1-[[2-O-ben zoyl-3,4-
O-isop r op ylid en e-6-O-(ter t-bu tyld im eth ylsilyl)-â-D-ga la c-
top yr a n osyl]oxy]-4-octa d ecen e (35). (2S,3R,4E)-2-Azido-
3-hydroxy-1-[[3,4-O-isopropylidene-6-O-(tert-butyldimethylsilyl)-
â-D-galactopyranosyl]oxy]-4-octadecene (34) (537 mg, 0.837
mmol) was reacted by the procedure used to synthesize
compound 12 and afforded the title compound (572 mg, 80%).
(2S,3R,4E)-2-(H exa d eca n oyla m in o)-3-(b en zoyloxy)-1-
[(2,3-d i-O-b en zoyl-6-O-p iva loyl-â-D-ga la ct op yr a n osyl)-
oxy]-4-octa d ecen e (28). (2S,3R,4E)-2-Azido-3-(benzoyloxy)-
1-[(2,3-di-O-benzoyl-6-O-pivaloyl-â-D-galactopyranosyl)oxy]-4-
octadecene (27) (0.250 g, 0.28 mmol) was reacted by the
procedure used to synthesize compound 10 and afforded the
title compound (0.092 g, 30%) as a white solid.
IR (CH2Cl2) νmax (cm-1): 3700-3400 (O-H, N-H), 3050,
2930, 2860 (C-H), 1730, 1670 (CdO). 1H NMR (400 MHz,
CDCl3) δ: 0.89 (6H, t, J ) 6.8 Hz, 2 × -CH3), 1.12-1.44 (46H,
m, -(CH2)13-, -(CH2)10-), 1.16 (9H, s, -OCOC(CH3)3), 1.58-1.68
(2H, m, -CH2-), 1.81 (2H, t, J ) 7.7 Hz, -NHCOCH2-), 2.01 (2H,
qa, J ) 6.9 Hz, dCH-CH2-), 3.63 (1H, dd, J ) 9.8, 3.8 Hz, H-1),
3.85 (1H, t, J ) 6.4 Hz, H-5′), 4.06 (1H, dd, J ) 11.4, 6.4 Hz,
H-6′), 4.17 (1H, dd, J ) 9.8, 3.0 Hz, H-1), 4.18 (1H, br s, H-4′),
4.28 (1H, dd, J ) 11.4, 6.5 Hz, H-6′), 4.44 (1H, m, H-2), 4.64
(1H, d, J ) 7.8 Hz, H-1′), 5.33 (1H, dd, J ) 10.4, 3.1 Hz, H-3′),
5.48 (1H, dd, J ) 15.3, 7.4 Hz, H-4), 5.58 (1H, br t, H-3), 5.70
(1H, dd, J ) 10.4, 7.8 Hz, H-2′), 5.85 (1H, dt, J ) 15.3, 6.9 Hz,
H-5), 7.36-7.59, 7.94-8.00, and 8.04-8.06 (16H, 3 sets of m,
aromatic H, -NH-).
(2S,3R,4E)-2-(H exa d eca n oyla m in o)-3-(b en zoyloxy)-1-
[[2,3-di-O-ben zoyl-6-O-pivaloyl-4-O-(sodiu m oxysu lfon yl)-
â-D-ga la ctop yr a n osyl]oxy]-4-octa d ecen e (29). (2S,3R,4E)-
2-(Hexadecanoylamino)-3-(benzoyloxy)-1-[(2,3-di-O-benzoyl-6-
O-pivaloyl-â-D-galactopyranosyl)oxy]-4-octadecene (28) (0.085
g, 0.078 mmol) was reacted by the procedure used to synthesize
compound 16 and afforded the title compound (0.072 g, 77%)
as a white solid.
IR (CH2Cl2) νmax (cm-1): 3700-3400 (N-H), 3050, 2930,
2860 (C-H), 1725, 1670 (CdO). 1H NMR (400 MHz, CDCl3)
δ: 0.88 (6H, br t, 2 × -CH3), 1.00 (9H, s, -OCOtBu), 1.03-1.37
(48H, m, -(CH2)13-, -(CH2)10-), 1.68 (2H, t, J ) 7.3 Hz, -NH-
COCH2-), 1.95 (2H, m, dCH-CH2-), 3.59, 3.95, 4.08-4.10, and
4.40-4.49 (1H, 1H, 2H, 2H, br d, br s, 2 sets of m, H-5′, H-6′,
H-2, H-1), 4.66 (1H, d, J ) 7.7 Hz, H-1′), 5.13 (1H, br s, H-4′),
5.35 (1H, br d, H-3′), 5.44 (1H, dd, J ) 15.1, 7.4 Hz, H-4), 5.55
(1H, t, J ) 7.2 Hz, H-3), 5.70 (1H, dd, J ) 10.0, 7.7 Hz, H-2′),
5.80 (1H, dt, J ) 15.1, 6.7 Hz, H-5), 7.35-7.42, 7.49-7.53, and
7.96-8.02 (16H, 3 sets of m, aromatic H, -NH-).
(2S,3R,4E)-2-(Hexa d eca n oyla m in o)-3-h yd r oxy-1-[[4-O-
(sod iu m oxysu lfon yl)-â-D-ga la ctop yr a n osyl]oxy]-4-octa -
d ecen e (30). (2S,3R,4E)-2-(Hexadecanoylamino)-3-(benzoy-
loxy)-1-[[2,3-di-O-benzoyl-6-O-pivaloyl-4-O-(sodium oxysulfonyl)-
â-D-galactopyranosyl]oxy]-4-octadecene (29) (0.070 g, 0.058
mmol) was reacted by the procedure used to synthesized
compound 11 and gave the title compound (12 mg, 26%) as a
beige solid.
IR (CH2Cl2) νmax (cm-1): 3700-3050 (N-H, O-H), 2930,
2850 (C-H), 1640 (CdO). 1H NMR (400 MHz, CDCl3) δ: 0.85
(6H, t, J ) 6.7 Hz, 2 × -CH3), 1.23-1.27 (46H, m, -(CH2)13-,
-(CH2)10-), 1.44 (2H, m, -CH2-), 1.92-2.04 (4H, m, -NHCOCH2-,
dCH-CH2-), 3.23 (1H, dd, J ) 9.3, 7.6 Hz, H-2′), 3.29-3.52
(5H, m, H-1, H-3′, H-5′, H-6′), 3.75 (1H, m, H-2), 3.89 (1H, br
t, H-3), 3.98 (1H, dd, J ) 9.9, 4.4 Hz, H-1), 4.05 (1H, d, J )
7.6 Hz, H-1′), 4.38 (1H, d, J ) 3.1 Hz, H-4′), 4.45 (1H, br s,
-OH), 4.69 (1H, d, J ) 6.1 Hz, -OH), 4.90 and 5.03 (2H, 2br s,
2 × -OH), 5.34 (1H, d, J ) 15.3, 7.1 Hz, H-4), 5.52 (1H, dt, J
) 15.3, 6.7 Hz, H-5), 7.51 (1H, d, J ) 9.0 Hz, -NH-). MS (CI,
NH3): 778 (M - Na)-.
Disu lfa ted Com p ou n d s. Syn th esis of (2S,3R,4E)-2-
(Hexadecan oylam in o)-3-h ydr oxy-1-[[2,6-bis-O-(sodiu m ox-
ysu lfon yl)-â-D-ga la ctop yr a n osyl]oxy]-4-octa d ecen e (32).
(2S,3R,4E)-2-(Hexadecan oylam in o)-3-(ben zoyloxy)-1-[[2,6-
bis-O-(sod iu m oxysu lfon yl)-â-D-ga la ctop yr a n osyl]oxy]-4-
octa d ecen e (31). (2S,3R,4E)-2-(Hexadecanoylamino)-3-(ben-
zoyloxy)-1-[(3,4-O-isopropylidene-â-D-galactopyranosyl)oxy]-4-
octadecene (19) (160 mg, 0.19 mmol) was reacted by the
procedure used to synthesize compounds 16 and 14 and gave
the title compound (115 mg, 60%) as a white solid.
IR (Nujol) νmax (cm-1): 3700-3100 (O-H, N-H), 2930, 2860
(C-H), 1715 (CdO ester), 1650 (CdO amide), 1270, 1010
(SdO). 1H NMR (400 MHz, DMSO-d6) δ: 0.85 (6H, t, J ) 6.8
Hz, 2 × -CH3), 1.21-1.44 (48H, br m, -(CH2)13-, -(CH2)11-), 1.97
1H NMR (200 MHz, CDCl3) δ: 0.09 (6H, s, -Si(CH3)2), 0.84-
0.94 (3H, m overlapping -C(CH3)3, -CH2-CH3), 0.90 (9H, s,
-C(CH3)3), 1.25 (22H, m, -(CH2)11-), 1.34 and 1.57 (2 × 3H, 2s,

Sulfatides as Potential Antiinflammatory Agents
J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 20 3245
(CH3)2-C-), 1.92 (2H, m, dCH-CH2-), 3.54 (1H, m, H-2), 3.86-
3.95 and 4.26-4.38 (7H, 2m, H-3′, H-4′, H-5′, H-6′, H-1), 4.51
(1H, d, J ) 8.1 Hz, H-1′), 5.22-5.78 (4H, m, H-3, H-4, H-5,
H-2′), 7.30-8.08 (10H, 2m, 2 × -C6H5).
(hexadecanoylamino)-1-[[2-O-benzoyl-4-O-acetyl-6-O-(tert-
butyldimethylsilyl)-â-D-galactopyranosyl]oxy]-4-octadecene (38)
(430 mg, 0.484 mmol) in tetrahydrofuran (40 mL) was treated
with tetrabutylammonium fluoride (1 M solution in tetrahy-
drofuran, 5 mL, 0.5 mmol) at -15 °C. The reaction mixture
was stirred for 36 h at 0 °C, then diluted with ethyl acetate,
and washed with water (4x), 10% aqueous sodium bicarbonate
solution (3x), and water with solid sodium bicarbonate (3x).
The organic layer was dried over anhydrous magnesium
sulfate, filtered, and concentrated. The resulting white solid
was used for the next reaction without further purification.
1H NMR (400 MHz, CDCl3) δ: 0.89 (6H, m, 2 × -CH3), 1.19-
1.41 (48H, m, -(CH2)11-, -(CH2)13-), 1.44 (2H, br s, 2 × -OH),
1.90 (2H, t, J ) 7.7 Hz, -CH2CONH-), 2.00 (2H, m, dCH-
CH2-), 2.26 (3H, s, CH3CO-), 3.41 (1H, dd, J ) 11.9, 5.8 Hz,
H-6′), 3.58 (1H, dd, J ) 11.9, 6.6 Hz, H-6′), 3.64-3.70 (2H, m,
H-5′, H-3′), 3.99 (1H, dd, J ) 10.0, 3.6 Hz, H-1), 4.03 (1H, dd,
J ) 10.0, 2.8 Hz, H-1), 4.47 (1H, m, H-2), 4.55 (1H, d, J ) 7.9
Hz, H-1′), 5.26 (1H, dd, J ) 10.0, 7.9 Hz, H-2′), 5.29 (1H, d, J
) 3.8 Hz, H-4′), 5.48 (1H, dd, J ) 15.3, 7.7 Hz, H-4), 5.60 (1H,
dd, J ) 7.7, 7.7 Hz, H-3), 5.71 (1H, d, J ) 9.3 Hz, -NH-), 5.87
(1H, dt, J ) 15.3, 6.8 Hz, H-5), 7.43-8.06 (10H, 3m, 2 × -C6H5).
(2S,3R,4E)-3-(Ben zoyloxy)-2-(h exa d eca n oyla m in o)-1-
[[2-O-ben zoyl-4-O-a cetyl-3,6-bis-O-(sod iu m oxysu lfon yl)-
â-D-ga la ctop yr a n osyl]oxy]-4-octa d ecen e (40). (2S,3R,4E)-
3-(Benzoyloxy)-2-(hexadecanoylamino)-1-[(2-O-benzoyl-4-O-
acetyl-â-D-galactopyranosyl)oxy]-4-octadecene (39) (100 mg,
0.105 mmol) was reacted by the procedure used to synthesize
compound 8 and afforded the title compound (94 mg, 77%) as
a white solid.
(2S,3R,4E)-3-(Ben zoyloxy)-2-(h exa d eca n oyla m in o)-1-
[[2-O-ben zoyl-3,4-O-isop r op ylid en e-6-O-(ter t-bu tyld im e-
th ylsilyl)-â-D-ga la ctop yr a n osyl]oxy]-4-octa d ecen e (36).
(2S,3R,4E)-2-azido-3-(benzoyloxy)-1-[[2-O-benzoyl-3,4-O-iso-
propylidene-6-O-(tert-butyldimethylsilyl)-â-D-galactopyranosyl]-
oxy]-4-octadecene (35) (572 mg, 0.673 mmol) was reacted by
the procedure used to synthesize compound 10 and afforded
the title compound (575 mg, 96%).
1H NMR (200 MHz, CDCl3) δ: 0.02 and 0.04 (2 × 3H, 2s,
-Si(CH3)2), 0.85-0.91 (6H, m overlapping -C(CH3)3, 2 × -CH2-
CH3), 0.86 (9H, s, -C(CH3)3), 1.13-1.38 (48H, m, -(CH2)11-,
-(CH2)13-), 1.34 and 1.60 (2 × 3H, 2s, (CH3)2-C-), 1.74 (2H, ap
t, J ) 7.5 Hz, dCH-CH2-), 1.98 (2H, m, -CH2CONH-), 3.57 (1H,
dd, J ) 3.9, 9.9 Hz, H-1), 3.69-3.89 (3H, m, H-6′, H-5′), 4.11
(1H, dd, J ) 3.1, 9.9 Hz, H-1), 4.25-4.36 (3H, m, H-2, H-3′,
H-4′), 4.41 (1H, d, J ) 8.1 Hz, H-1′), 5.19 (1H, dd, J ) 7.1, 8.0
Hz, H-3), 5.40-5.52 (2H, m, H-2′, H-4), 5.71 (1H, d overlapping
H-5, J ) 9.3 Hz, -NH-), 5.77 (1H, dt, J ) 14.3, 6.9 Hz, H-5),
7.39 (10H, 3m, 2 × -C6H5).
(2S,3R,4E)-3-(Ben zoyloxy)-2-(h exa d eca n oyla m in o)-1-
[[2-O-b en zoyl-6-O-(ter t-b u t yld im et h ylsilyl)-â-D-ga la ct o-
pyr an osyl]oxy]-4-octadecen e (37). Trifluoroacetic acid (90%,
≈4 mL) was added to a stirred solution of (2S,3R,4E)-3-
(benzoyloxy)-2-(hexadecanoylamino)-1-[[2-O-benzoyl-3,4-O-iso-
propylidene-6-O-(tert-butyldimethylsilyl)-â-D-galactopyranosyl]-
oxy]-4-octadecene (36) (575 mg, 0.649 mmol) in dichloromethane
(75 mL) at 22 °C. The reaction mixture was stirred at 22 °C
and monitored by TLC. The solvents were evaporated, and
the residue was dissolved in ethyl acetate. The organic layer
was washed with a 10% aqueous solution of sodium bicarbon-
ate, water, and brine, dried over anhydrous magnesium
sulfate, filtered, and concentrated.
The residue was dissolved in pyridine (25 mL) and treated
with tert-butyldimethylsilyl chloride (425 mg, 2.82 mmol) at
-15 °C. The reaction mixture was stirred at -15 °C overnight;
then methanol was added. The mixture was stirred again for
2 more hours at 22 °C. The reaction mixture was then poured
into water and diluted with ethyl acetate (150 mL). The
organic layer was washed with water (5x) and brine, dried over
anhydrous magnesium sulfate, filtered, and concentrated. The
residue was purified by silica gel chromatography and afforded
the title compound.
IR (Nujol) νmax (cm-1): 3700-3150 (N-H), 2730, 2860 (C-
H), 1725 (CdO ester), 1645 (CdO amide). 1H NMR (400 MHz,
DMSO-d6) δ: 0.85 (6H, m, 2 × -CH3), 1.10-1.40 (48H, m,
-(CH2)11-, -(CH2)13-), 1.74 (2H, m, dCH-CH2-), 1.94 (2H, m,
-CH2CONH-), 2.08 (3H, s, -CH3CO-), 3.47 (1H, dd, J ) 9.3,
7.8 Hz, H-1), 3.61 (1H, dd, J ) 11.1, 7.7 Hz, H-6′), 3.74-3.78
(2H, m, H-1, H-6′), 4.07 (1H, dd, J ) 7.7, 3.7 Hz, H-5′), 4.27
(1H, m, H-2), 4.57 (1H, dd, J ) 10.3, 3.3 Hz, H-3′), 4.72 (1H,
d, J ) 8.0 Hz, H-1′), 5.09 (1H, dd, J ) 10.3, 8.0 Hz, H-2′), 5.29
(1H, dd, J ) 6.9, 4.3 Hz, H-3), 5.34-5.43 (2H, m, H-4, H-5),
5.46 (1H, d, J ) 3.3 Hz, H-4′), 7.41-7.99 (10H, 4m, 2 × -C6H5),
7.67 (1H, d, J ) 9.0 Hz, -NH-).
(2S,3R,4E)-3-Hyd r oxy-2-(h exa d eca n oyla m in o)-1-[[3,6-
bis-O-(sod iu m oxysu lfon yl)-â-D-ga la ctop yr a n osyl]oxy]-4-
octa d ecen e (41). (2S,3R,4E)-3-(Benzoyloxy)-2-(hexadecanoy-
lamino)-1-[[2-O-benzoyl-4-O-acetyl-3,6-bis-O-(sodium oxysul-
fonyl)-â-D-galactopyranosyl]oxy]-4-octadecene (40) (100 mg,
0.087 mmol) was reacted by the procedure used to synthesize
compound 11 and afforded the title compound (43 mg, 55%)
as a white solid.
1H NMR (400 MHz, CDCl3) δ: 0.01 and 0.03 (2 × 3H,
2s, -Si(CH3)2), 0.86 (9H, s overlapping -CH3, -C(CH3)3), 0.86-
0.90 (6H, m, 2 × -CH2-CH3), 1.10-1.26 (46H, m, -(CH2)10-,
-(CH2)13-), 1.64 (2H, m, -CH2-), 1.78 (2H, t, J ) 7.6 Hz, dCH-
CH2-), 1.98 (2H, m, -CH2CONH-), 3.48 (1H, t, H-5′), 3.60 (1H,
dd, J ) 9.4, 3.4 Hz, H-1), 3.66 (1H, dd, J ) 10.5, 4.3 Hz, H-6′),
3.75-3.81 (2H, m, H-6′, H-3′), 4.07 (1H, d, J ) 3.1 Hz, H-4′),
4.13 (1H, d, H-1), 4.42 (1H, m, H-2), 4.48 (1H, d, J ) 7.8 Hz,
H-1′), 5.25 (1H, dd, J ) 8.8, 8.8 Hz, H-2′), 5.44 (1H, dd, J )
15.3, 7.6 Hz, H-4), 5.53 (1H, dd, J ) 7.4, 7.4 Hz, H-3), 5.77
(1H, d, J ) 9.2 Hz, -NH-), 5.84 (1H, dt, J ) 15.3, 6.7 Hz, H-5),
7.43-8.06 (10H, 3m, 2 × -C6H5).
IR (Nujol) νmax (cm-1): 3700-3100 (O-H, N-H), 2730, 2860
(C-H), 1640 (CdO amide). 1H NMR (400 MHz, DMSO-d6) δ:
0.84 (6H, t, J ) 6.7 Hz, 2 × -CH3), 1.22-1.43 (48H, m, -(CH2)11-,
-(CH2)13-), 1.91 (2H, m, dCH-CH2-), 2.01 (2H, t, J ) 7.3 Hz,
-CH2CONH-), 3.41-3.50 (2H, m, H-1, H-6′), 3.58 (1H, ∼t, J )
5.9 Hz, H-5′), 3.73-3.87 (5H, m, H-1, H-6′, H-2′, H-4′, H-3),
3.94 (1H, dd overlapping H-2, J ) 9.8, 3.3 Hz, H-3′), 3.93-
3.96 (1H, m, H-2), 4.15 (1H, d, J ) 7.7 Hz, H-1′), 4.56, 4.86,
5.09 (3H, 3s, 3 × -OH), 5.34 (1H, dd, J ) 15.3, 6.9 Hz, H-4),
5.51 (1H, dt, J ) 15.3, 6.6 Hz, H-5), 7.48 (1H, d, J ) 8.9 Hz,
-NH-). MS (CI, NH3): 903 (M - H)-.
Syn t h esis of (2S,3R,4E)-3-H yd r oxy-2-(h exa d eca n oyl-
a m in o)-1-[[4,6-bis-O-(sod iu m oxysu lfon yl)-â-D-ga la ctop y-
r a n osyl]oxy]-4-oct a d ecen e (46). (2S,3R,4E)-2-Azid o-3-
(ben zoyloxy)-1-[(4,6-O-ben zyliden e-â-D-galactopyr an osyl)-
oxy]-4-octa d ecen e (42). Benzaldehyde (5 mL) was added to
a solution of (2S,3R,4E)-2-azido-3-(benzoyloxy)-1-(â-D-galac-
topyranosyloxy)-4-octadecene (6) (450 mg, 0.76 mmol) in formic
acid (5 mL) at 22 °C and under argon. This mixture was
stirred for 0.75 h, then cooled down to 5 °C, and diluted with
ethyl acetate (25 mL) and water (10 mL). The two-phase
solution was neutralized by adding solid sodium bicarbonate
and a solution of sodium bicarbonate (1 M). The aqueous
phase was then extracted with ethyl acetate (2 × 25 mL). The
combined organic layers were washed with a cold solution of
sodium bicarbonate (1 M, 25 mL), water (25 mL), and brine
(2S,3R,4E)-3-(Ben zoyloxy)-2-(h exa d eca n oyla m in o)-1-
[[2-O-ben zoyl-4-O-a cetyl-6-O-(ter t)-bu tyld im eth ylsilyl)-â-
D-ga la ctop yr a n osyl]oxy]-4-octa d ecen e (38). The title com-
pound was prepared from 37 according to the procedure used
to synthesize compound 15.
1H NMR (400 MHz, DMSO-d6) δ: -0.03 and -0.01 (2 × 3H,
2s, -Si(CH3)2), 0.82 (9H, s overlapping -CH3, -C(CH3)3), 0.82-
0.85 (6H, m, 2 × -CH2-CH3), 1.13-1.35 (48H, m, -(CH2)11-,
-(CH2)13-), 1.76-1.94 (4H, m, dCH-CH2-, -CH2CONH-), 2.07
(3H, s, CH3CO-), 3.47-3.56 (3H, m, H-6′, H-1), 3.72 (1H, dd,
J ) 10.0, 6.5 Hz, H-1), 3.81 (1H, t, H-5′), 3.94-3.99 (1H, m,
H-3′), 4.29 (1H, m, H-2), 4.62 (1H, d, J ) 8.0 Hz, H-1′), 5.01
(1H, dd, J ) 9.9, 8.0 Hz, H-2′), 5.24 (1H, d, J ) 3.4 Hz, H-4′),
5.30 (1H, dd, J ) 7.2, 4.7 Hz, H-3), 5.37-5.43 (2H, m, H-4,
-NH-), 5.48 (1H, dt, J ) 15.2, 6.3 Hz, H-5), 7.44-7.95 (10H,
4m, 2 × -C6H5).
(2S,3R,4E)-3-(Ben zoyloxy)-2-(h exa d eca n oyla m in o)-1-
[(2-O-ben zoyl-4-O-a cetyl-â-D-ga la ctop yr a n osyl)oxy]-4-oc-
t a d ecen e (39). A solution of (2S,3R,4E)-3-(benzoyloxy)-2-

3246 J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 20
Marinier et al.
(25 mL), dried over anhydrous magnesium sulfate, filtered,
and concentrated. The residue was purified by silica gel
chromatography (20 g, 10% acetone/ethyl acetate) and afforded
the title compound (233 mg, 45%) as a pale yellow oil.
IR (CH2Cl2) νmax (cm-1): 3580 (O-H), 3060, 2930, 2860 (C-
H), 2110 (-N3), 1720 (CdO), 1265 (C-O). 1H NMR (400 MHz,
CDCl3) δ: 0.89 (3H, t, J ) 6.9 Hz, -CH3), 1.25 (20H, br s,
-(CH2)10-), 1.39 (2H, qi, J ) 6.9 Hz, -CH2-CH3), 2.09 (2H, m,
dCH-CH2-), 3.49 (1H, d, J ) 0.9 Hz, H-5′), 3.69-3.74 (2H, m,
-CHN3-, H-1), 3.80 (1H, dd, J ) 9.6, 7.6 Hz, H-1), 3.99-4.05
(2H, m, H-2′, H-3′), 4.08 (1H, dd, J ) 12.5, 1.8 Hz, H-6′), 4.23
(1H, dd, J ) 3.5, 0.9 Hz, H-4′), 4.33 (1H, dd, overlapping H-1′,
J ) 12.5, 1.3 Hz, H-6′), 4.35 (1H, d, J ) 7.5 Hz, H-1′), 5.56
(1H, s, -O-CH-O-), 5.61 (1H, ddt, J ) 15.2, 8.0, 1.2 Hz, H-4),
5.69 (1H, dd, J ) 8.0, 4.0 Hz, H-3), 5.97 (1H, dt, J ) 15.2, 6.8
Hz, H-5), 7.34-8.12 (10H, 4m, 2 × -C6H5).
NMR (400 MHz, DMSO-d6) δ: 0.83 (6H, 2t, J ) 6.9 Hz, 2 ×
-CH3), 1.14-1.50 (48H, m, -(CH2)11-, -(CH2)13-), 1.75-2.00 (4H,
m, -COCH2-, dCH-CH2-), 3.57-3.61 (1H, m, H-5′), 3.82-3.88
and 4.08-4.12 (2 × 2H, 2m, H-6′, H-1), 4.07-4.13 (1H, m, H-2),
4.66 (1H, d, J ) 3.3 Hz, H-4′), 4.80 (1H, d, J ) 7.7 Hz, H-1′),
5.27-5.54 (5H, m, H-3, H-3′, H-2′, H-5, H-4), 7.35-7.88 (15H,
3m, 3 × -C6H5), 7.76 (1H, d, J ) 8.9 Hz, Nˇ H-).
(2S,3R,4E)-3-Hyd r oxy-2-(h exa d eca n oyla m in o)-1-[[4,6-
bis-O-(sod iu m oxysu lfon yl)-â-D-ga la ctop yr a n osyl]oxy]-4-
octa d ecen e (46). (2S,3R,4E)-3-(Benzoyloxy)-2-(hexadecano-
ylamino)-1-[[2,3-di-O-benzoyl-4,6-bis-O-(sodium oxysulfonyl)-
â-D-galactopyranosyl]oxy]-4-octadecene (45) (1.0 g, 0.82 mmol)
was reacted by the procedure used to synthesize compound
11 and afforded the title compound (212 mg, 20%) as an off-
white solid.
IR (Nujol) νmax (cm-1): 3700-3100 (O-H, N-H), 2930, 2860
(C-H), 1640 (CdO amide). 1H NMR (400 MHz, DMSO-d6) δ:
0.84 (6H, t, J ) 6.7 Hz, 2 × -CH3), 1.05-1.26 (46H, br s,
-(CH2)10-, -(CH2)13-), 1.43 (2 H, m, -CH2-), 1.91 (2H, m, dCH-
CH2-), 2.02 (2H, t, J ) 7.4 Hz, -CH2-CONH-), 3.21 (1H, dd, J
) 9.6, 7.7 Hz, H-2′), 3.36-3.41 (2H, m, H-3′, H-5′), 3.67-3.78
(3H, m, H-6′, H-1, H-2), 3.83-3.95 (2H, m, H-1, H-3), 4.00 (1H,
dd, J ) 9.8, 4.9 Hz, H-6′), 4.05 (1H, d, J ) 7.7 Hz, H-1′), 4.33
(1H, d, J ) 3.0 Hz, H-4′), 5.30 (1H, dd, J ) 15.3, 7.2 Hz, H-4),
5.51 (1H, dt, J ) 15.3, 6.6 Hz, H-5), 7.54 (1H, d, J ) 9.1 Hz,
-NH-).
Syn t h esis of (2S,3R,4E)-3-H yd r oxy-2-(h exa d eca n oyl-
a m in o)-1-[[2,3-bis-O-(sod iu m oxysu lfon yl)-â-D-ga la ctop y-
r a n osyl]oxy]-4-octa d ecen e (50). (2S,3R,4E)-3-(Ben zoyl-
oxy)-2-(h exa d eca n oyla m in o)-1-[(4,6-O-b en zylid en e-â-D-
ga la ctop yr a n osyl)oxy]-4-octa d ecen e (47). (2S,3R,4E)-2-
Azido-3-(benzoyloxy)-1-[(4,6-O-benzylidene-â-D-galactopyrano-
syl)oxy]-4-octadecene (42) (235 mg, 0.34 mmol) was reacted
by the procedure used to synthesize compound 10 and afforded
the title compound (230 mg, 76%) as a white solid.
(2S,3R,4E)-3-(Ben zoyloxy)-2-(h exa d eca n oyla m in o)-1-
[(2,3-d i-O-ben zoyl-4,6-O-ben zylid en e-â-D-ga la ctop yr a n o-
syl)oxy]-4-octa d ecen e (43). (2S,3R,4E)-2-Azido-3-(benzoy-
loxy)-1-[(2,3-di-O-benzoyl-4,6-O-benzylidene-â-D-ga la cto-
pyranosyl)oxy]-4-octadecene (25) (250 mg, 0.28 mmol) was
reacted by the procedure used to synthesize compound 10 and
afforded the title compound (266 mg, 86%) as a white solid.
IR (CH2Cl2) νmax (cm-1): 3060, 2930, 2860 (C-H), 1725 (CdO
ester), 1675 (CdO amide), 1265 (C-O). 1H NMR (400 MHz,
CDCl3) δ: 0.89 (6H, t, J ) 6.8 Hz, 2 × -CH3), 1.14-1.43 (48H,
m, -(CH2)11-, -(CH2)13-), 1.83 (2H, t, J ) 7.6 Hz, -COCH2-), 1.99
(2H, m, dCH-CH2-), 3.67 (1H, br s, H-5′), 3.81 (1H, dd, J )
10.4, 4.1 Hz, H-1), 4.11 (1H, d, J ) 12.4, 1.4 Hz, H-6′), 4.17
(1H, dd, J ) 10.4, 4.0 Hz, H-1), 4.30 (1H, dd, J ) 12.4, 1.1 Hz,
H-6′), 4.42-4.47 (1H, m, H-2), 4.58 (1H, d, J ) 3.5 Hz, H-4′),
4.74 (1H, d, J ) 8.0 Hz, H-1′), 5.38 (1H, dd, J ) 10.4, 3.5 Hz,
H-3′), 5.50 (1H, dd, J ) 15.3, 7.0 Hz, H-4), 5.55 (1H, s, -O-
CH-O-), 5.60 (1H, dd, J ) 7.0 Hz, H-3), 5.77-5.87 (1H, m,
overlapping H-2′, H-5), 5.85 (1H, dd, J ) 10.4, 8.0 Hz, H-2′),
7.35-7.99 (20 H, 4m, 4 × -C6H5), 8.04 (1H, d, J ) 8.5 Hz,
-NH-).
(2S,3R,4E)-3-(Ben zoyloxy)-2-(h exa d eca n oyla m in o)-1-
[(2,3-d i-O-b e n zoyl-â-D-ga la ct op yr a n osyl)oxy]-4-oct a -
d ecen e (44). Trifluoroacetic acid (90%, 0.5 mL) was added
to a stirred solution of (2S,3R,4E)-3-(benzoyloxy)-2-(hexade-
canoylamino)-1-[(2,3-di-O-benzoyl-4,6-O-benzylidene-â-D-galac-
topyranosyl)oxy]-4-octadecene (43) (258 mg, 0.23 mmol) in
dichloromethane (15 mL) at 5 °C. The mixture was stirred
for 0.5 h at 5 °C and at 22 °C for 1 h. Trifluoroacetic acid
(same quantity) was added again, and the reaction mixture
was stirred for 1 more hour at 22 °C. The mixture was diluted
with ethyl acetate (30 mL) and washed with a cold aqueous
solution of sodium bicarbonate (1 M, 2 × 15 mL), water (2 ×
15 mL), and brine (15 mL). The organic layer was dried over
anhydrous magnesium sulfate, filtered, and concentrated. The
residue was purified by silica gel chromatography (15 g, 0-60%
ethyl acetate/hexane) and afforded the title compound (193 mg,
83%) as a white solid.
IR (CH2Cl2) νmax (cm-1): 3055, 2930, 2860 (C-H), 1720 (CdO
ester), 1640 (CdO amide). 1H NMR (400 MHz, CDCl3) δ: 0.88
(6H, t, J ) 6.3 Hz, 2 × -CH3), 1.24 (44H, br s, -(CH2)10-,
-(CH2)12-), 1.60 (4H, br s, 2 × -CH2-), 2.03 (2H, m, dCH-CH2-),
2.19 (2H, t, J ) 7.5 Hz, -CH2CONH-), 3.48-4.34 (9H, m, H-1,
H-1′, H-2′, H-3′, H-4′, H-5′, H-6′), 4.52 (1H, m, H-2), 5.44-
5.63 (3H, m, H-3, H-4, -O-CH-O-), 5.88 (1H, dt, J ) 14.8, 6.7
Hz, H-5), 6.21 (1H, d, J ) 9.2 Hz, -NH-), 7.34-8.05 (10H, 2m,
2 × -C6H5).
(2S,3R,4E)-3-(Ben zoyloxy)-2-(h exa d eca n oyla m in o)-1-
[[2,3-bis-O-(sod iu m oxysu lfon yl)-4,6-O-ben zylid en e-â-D-
ga la ctop yr a n osyl]oxy]-4-octa d ecen e (48). (2S,3R,4E)-3-
(Benzoyloxy)-2-(hexadecanoylamino)-1-[(4,6-O-benzylidene-â-
D-galactopyranosyl)oxy]-4-octadecene (47) (66 mg, 0.074 mmol)
was reacted by the procedure used to synthesize compound
16 and afforded the title compound (70 mg, 86%) as a white
solid.
IR (CH2Cl2) νmax (cm-1): 3700-3150 (N-H), 2925, 2860 (C-
H), 1725 (CdO ester), 1650 (CdO amide), 1265 (C-O, SdO).
1H NMR (400 MHz, DMSO-d6) δ: 0.84 (6H, t, J ) 6.9 Hz, 2 ×
-CH3), 1.18-1.27 (46H, br s, -(CH2)11-, -(CH2)12-), 1.42 (2H, m,
-CH2-), 1.93 (2H, m, dCH-CH2-), 2.00-2.19 (2H, m, -CH2-
CONH-), 3.40 (1H, dd, J ) 10.0, 3.5 Hz, H-1), 3.50 (1H, s, H-5′),
3.86 (1H, d, J AB ) 11.1 Hz, H-6′), 3.96 (1H, d, J AB ) 11.1 Hz,
H-6′), 4.09-4.14 (1H, m overlapping H-1, H-3′), 4.13 (1H, dd,
J ) 10.0, 3.3 Hz, H-1), 4.25 (1H, m, H-2), 4.31 (1H, dd, J )
9.9, 7.8 Hz, H-2′), 4.37 (1H, d, J ) 7.8 Hz, H-1′), 4.57 (1H, d,
J ) 3.3 Hz, H-4′), 5.38-5.48 (3H, m, -O-CH-O-, H-3, H-4), 5.71
(1H, dt, J ) 14.5, 7.7 Hz, H-5), 7.30-7.99 (10H, 4m, 2 × -C6H5),
8.30 (1H, d, J ) 9.0 Hz, -NH-).
IR (CH2Cl2) νmax (cm-1): 3060, 2930, 2860 (C-H), 1730 (CdO
ester), 1670 (CdO amide), 1265 (C-O). 1H NMR (400 MHz,
CDCl3) δ: 0.89 (6H, t, J ) 6.8 Hz, 2 × -CH3), 1.25-1.57 (48H,
m, -(CH2)11-, -(CH2)13-), 1.97-2.04 (4H, m, -COCH2-, dCH-
CH2-), 3.53 (1H, t overlapping -OH-6′, J ) 3.0 Hz, H-5′), 3.53-
3.58 (1H, m, -OH-6′), 3.67-3.71 (2H, m, H-1, H-6′), 3.86 (1H,
dd, J ) 12.6, 3.5 Hz, H-6′), 4.01 (1H, dd, J ) 9.7, 1.8 Hz, H-1),
4.25 (1H, br s, H-4′), 4.49-4.54 (1H, m, H-2), 4.66 (1H, d, J )
7.8 Hz, H-1′), 5.27 (1H, dd, J ) 10.4, 2.9 Hz, H-3′), 5.51 (1H,
dd, J ) 15.4, 8.2 Hz, H-4), 5.73-5.86 (2H, m, H-3, H-2′), 5.96
(1H, dt, J ) 15.3, 7.0 Hz, H-5), 7.36-8.07 (15H, 5m, 3 × -C6H5),
8.06 (1H, d, J ) 8.4 Hz, -NH-).
(2S,3R,4E)-3-(Ben zoyloxy)-2-(h exa d eca n oyla m in o)-1-
[[2,3-bis-O-(sod iu m oxysu lfon yl)-â-D-ga la ctop yr a n osyl]-
oxy]-4-octa d ecen e (49). (2S,3R,4E)-3-(Benzoyloxy)-2-(hexa-
decanoylamino)-1-[[2,3-bis-O-(sodium oxysulfonyl)-4,6-O-ben-
zylidene-â-D-galactopyranosyl]oxy]-4-octadecene (48) (140 mg,
0.128 mmol) was reacted by the procedure used to synthesize
compound 44 and afforded the title compound (70.5 mg, 55%)
as a white solid.
(2S,3R,4E)-3-(Ben zoyloxy)-2-(h exa d eca n oyla m in o)-1-
[[2,3-d i-O-ben zoyl-4,6-bis-O-(sod iu m oxysu lfon yl)-â-D-ga -
la ct op yr a n osyl]oxy]-4-oct a d ecen e (45). (2S,3R,4E)-3-
(Benzoyloxy)-2-(hexadecanoylamino)-1-[(2,3-di-O-benzoyl-â-D-
galactopyranosyl)oxy]-4-octadecene (44) (182 mg, 0.18 mmol)
was reacted by the procedure used to synthesize compound
16 and afforded the title compound (117 mg, 54%) as a white
solid.
IR (Nujol) νmax (cm-1): 3700-3150 (O-H, N-H), 2920, 2855
(C-H), 1720 (CdO ester), 1650 (CdO amide), 1265 (C-O,
SdO). 1H NMR (400 MHz, DMSO-d6) δ: 0.82 (6H, t, J ) 6.9
Hz, 2 × -CH3), 1.18-1.24 (46H, m, -(CH2)11-, -(CH2)12-), 1.41
IR (Nujol) νmax (cm-1): 3700-3200 (O-H), 2930, 2860 (C-
H), 1725 (CdO ester), 1650 (CdO amide), 1460 (SdO). 1H

Sulfatides as Potential Antiinflammatory Agents
J ournal of Medicinal Chemistry, 1997, Vol. 40, No. 20 3247
M. Sulfated Glycolipids are Ligands for a Lymphocyte Homing
Receptor, L-Selectin (LECAM-1), Binding Epitope in Sulfated
Sugar Chain. Biochem. Biophys. Res. Commun. 1993, 190, 426-
434. (c) Todderud, G.; Alford, J .; Millsap, K. A.; Aruffo, A.;
Tramposch, K. M. PMN Binding to P-Selectin is Inhibited by
Sulfatide. J . Leukocyte Biol. 1992, 52, 85.
(2H, m, -CH2-), 1.92 (2H, m, dCH-CH2-), 1.98-2.16 (2H, m,
-CH2CONH-), 3.27-3.33 (3H, m, H-1, H-5′, H-6′), 3.41 (1H,
dd, J ) 8.3, 4.4 Hz, H-6′) 3.90 (1H, m, H-2), 4.06 (1H, d, J )
8.3 Hz, H-1), 4.19-4.22 (4H, m, H-1′, H-2′, H-3′, H-4′), 4.51
(2H, br s, 2 × -OH), 5.33 (1H, t, J ) 7.6 Hz, H-3), 5.38 (1H,
dd, J ) 14.6, 7.6 Hz, H-4), 5.69 (1H, dt, J ) 14.6, 7.0 Hz, H-5),
7.48-7.97 (5H, 3m, -C6H5), 8.37 (1H, d, J ) 9.2 Hz, -NH-).
MS (CI, NH3): 962 (M - 2Na + H)-.
(2S,3R,4E)-3-Hyd r oxy-2-(h exa d eca n oyla m in o)-1-[[2,3-
bis-O-(sod iu m oxysu lfon yl)-â-D-ga la ctop yr a n osyl]oxy]-4-
octa d ecen e (50). A solution of (2S,3R,4E)-3-(benzoyloxy)-2-
(hexadecanoylamino)-1-[[2,3-bis-O-(sodium oxysulfonyl)-â-D-
galactopyranosyl]oxy]-4-octadecene (49) (46 mg, 0.045 mmol)
was reacted by the procedure used to synthesize compound
11 and afforded the title compound (26 mg, 64%) as a white
solid.
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Domains. Nature 1994, 367, 532-538.
IR (Nujol) νmax (cm-1): 3700-3150 (O-H, N-H), 2925, 2855
(C-H), 1650 (CdO amide). 1H NMR (400 MHz, DMSO-d6) δ:
0.84 (6H, t, J ) 6.8 Hz, 2 × -CH3), 1.22-1.46 (48H, m, -(CH2)11-,
-(CH2)13-), 1.88 (2H, m, dCH-CH2-), 1.93-2.11 (2H, m, -CH2-
CONH-), 3.09 (1H, dd, J ) 9.2, 2.7 Hz, H-1), 3.35 (1H, dd, t, J
) 6.2 Hz, H-5′), 3.45 (1H, dd, J ) 10.8, 6.2 Hz, H-6′), 3.51
(1H, dd, J ) 10.8, 6.2 Hz, H-6′), 3.67 (1H, br t, H-3), 3.84-
3.92 (2H, m, H-2, H-1), 4.11 (1H, d, J ) 3.9 Hz, -OH), 4.18-
4.25 (4H, m, H-1′, H-2′, H-3′, H-4′), 4.62 (1H, t, J ) 5.7 Hz,
-OH-6′), 4.77 (1H, d, J ) 5.8 Hz, -OH), 5.29 (1H, dd, J ) 15.4,
7.1 Hz, H-4), 5.45 (1H, dt, J ) 15.4, 6.6 Hz, H-5), 8.06 (1H, d,
J ) 9.4 Hz, -NH-).
Ack n ow led gm en t. The authors offer their sincere
appreciation to Dr. Stephen Klohr from BMS-PRI,
Wallingford, CT, for performing mass spectral analysis.
We would also like to acknowledge the contributions of
Mr. Gilles Lacasse, Dr. Christian Cote´, Ms. Danielle
Sirard, and Chantale J ean from BMS-PRI, Candiac,
Quebec, Analytical Department, for providing HPLC
analysis.
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J M9606960