Novel mimics of sialyl Lewis X: Design, synthesis and biological activity of a series of 2- and 3-malonate substituted galactoconjugates

Article abstract of DOI:10.1016/S0968-0896(01)00015-3

A series of potent inhibitors of P-selectin as potential antiinflammatory agents is reported. These compounds are derivatives of galactocerebrosides bearing a malonate side chain in positions 2 and 3 of the galactose moiety. Based on the binding mode of sialyl Lewis X, the two acidic groups of the malonate are designed to form ionic interactions with two important lysines in the active site of P-selectin, Lys113 and Lys111. On the other hand the 4- and 6-hydroxy groups on the galactosering are arranged to chelate the calcium ion in the P-selectin active site. The synthesis and the biological activity of this series of compounds are described. Lead compounds having a greater potency than sialyl lewis X are identified. Copyright

Full text of DOI:10.1016/S0968-0896(01)00015-3

Bioorganic & Medicinal Chemistry 9 (2001) 1395–1427
Novel Mimics of Sialyl Lewis X:
Design, Synthesis and Biological Activity of a Series of
2- and 3-Malonate Substituted Galactoconjugatesy
Anne Marinier,a,* Alain Martel,a Carol Bachand,a Serge Plamondon,a Brigitte Turmel,a
Jean-Paul Daris,a Jacques Banville,a Philippe Lapointe,a Carl Ouellet,a Pierre Dextraze,a
Marcel Menard,a John J. K. Wright,b Julie Alford,c Debbie Lee,c Paul Stanley,c
Xina Nair,c Gordon Todderudc and Kenneth M. Tramposchc
aDepartment of Chemistry, Bristol-Myers Squibb Pharmaceutical Research Institute, 100, blvd. de l’Industrie, Candiac, Canada J5R 1J1
bBristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, CT, USA
cDermatological Disease, Bristol-Myers Squibb Pharmaceutical Research Institute, Buffalo, NY, USA
Received 23 August 2000; accepted 11 January 2001
Abstract—A series of potent inhibitors of P-selectin as potential anti-inflammatory agents is reported. These compounds are deri-
vatives of galactocerebrosides bearing a malonate side chain in positions 2 and 3 of the galactose moiety. Based on the binding
mode of sialyl Lewis X, the two acidic groups of the malonate are designed to form ionic interactions with two important lysines in
the active site of P-selectin, Lys113 and Lys111. On the other hand, the 4- and 6-hydroxy groups on the galactose ring are arranged
to chelate the calcium ion in the P-selectin active site. The synthesis and the biological activity of this series of compounds are
described. Lead compounds having a greater potency than sialyl Lewis X are identified. # 2001 Elsevier Science Ltd. All rights
reserved.
Introduction
P-selectin is found in platelets as well as in endothelial
cells activated by thrombin and other mediators. E-sel-
ectin is expressed by vascular endothelial cells after
stimulation by endotoxins and cytokines associated with
infection and inflammation and interact with lympho-
cytes and neutrophils. L-selectin is present on the sur-
face of most lymphocytes and neutrophils and supports
their adhesion to endothelium.3
The selectins are a highly conserved gene family
having a common structural organization and are
implicated in the inflammatory response to tissue
injury and infection. The first event in this inflam-
matory cascade is the attachment of leukocytes to
cells of the vascular endothelium proximal to the site
of trauma. The selectins support this adhesion by
interacting with oligosaccharides of the sialyl Lewis X
(1, Fig. 1) (sLex) family presented by glycoproteins or
glycolipids on the surface of leukocytes.1 As expected
for a protein–carbohydrate interaction, the binding of
selectins to sLex is of low-affinity (0.5–1.0 mM).2 There-
fore, it has been postulated that in order to tether cir-
culating leukocytes in vivo, there must be multivalent
interactions between clustered patches of sLex and the
protein.
The selectins are transmembrane glycoproteins contain-
ing an amino-terminal C-type lectin domain which is
believed to be the site of carbohydrate recognition. The
epidermal growth factor (EGF) repeat and a variable
number of complement regulatory-like repeats con-
stitute the main structural difference between E-, P-, and
L-selectins.1
On the basis of the homology with mannose binding
protein, a C-type lectin whose structural coordinates
have been generated by X-ray,4 models for the lectin
domain of E- and P-selectin have been derived. Various
point mutants of E- and P-selectins were prepared and
allowed the identification of a small groove located at
*Corresponding author. Fax: +1-450-444-4142; e-mail: anne.-
marinier@ bms.com
yThis work has been presented in part at the 212th ACS National
Meeting, Orlando, FA, August 1996.
0968-0896/01/$ - see front matter # 2001 Elsevier Science Ltd. All rights reserved.
PII: S0968-0896(01)00015-3

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A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
the surface of the protein as a potential binding site.5À7
This pocket is in the vicinity of a calcium cation found
in all selectins as well as in the mannose binding protein.
An analysis of the critical residues for binding to P- and
E-selectin has revealed the importance of the conserved
Lys 113 for P-selectin and both conserved Lys 111 and
Lys 113 for E-selectin, located in this shallow groove.5À8
The recently published crystal structure of the lectin and
EGF domains of E-selectin has corroborated the
previous models and provides useful structural details to
elucidate the recognition of sLex epitope by the
selectins.9
mouse models of inflammation when administered sub-
cutaneously. It is in light of these results that we report
our efforts directed to the identification of novel glyco-
tope inhibitors of P-selectin adhesion. Glycomimetics of
sialyl Lewis X were designed and their synthesis and
biological affinity to P-selectin are reported in this
paper.
Design Considerations
A convergent strategy was used to design the series of
malonate derivatives reported here. In a first approach,
the minimum structural requirements previously descri-
bed for sLex binding to selectins were patterned. The
second structural element used was that of sulfatide (3,
Fig. 4), which was shown to bind to the P-selectin
receptor with an avidity that is equivalent to that of the
myeloid HL-60 cell line.20 In the case of the sulfatides,
the negatively charged sulfate group is suspected to
make an ionic interaction with Lys 113 and/or Lys 111
in P-selectin as for the carboxylate group of the sialic
acid moiety in sLex. Moreover, since glycosphingolipids
can coaggregate in patches on cell surfaces,21 this nat-
ural mechanism could generate specific recognition as it
is presumed for the natural ligand. These known struc-
tural features required for binding to selectins were
consequently combined in one simpler saccharide motif.
The sialyl Lewis X determinant binds to E-, P-, and
L-selectins through calcium dependent interactions.10
Although the role of calcium in ligand interactions has
not been unambiguously established, various SARs of
sLex-containing oligosaccharides11À15 suggests that the
critical fucosyl residue in sLex interacts with the protein
similarly to the binding of mannose with the mannose
binding protein (MBP). The crystal structure of this
mannose–MBP complex has revealed that the key fea-
ture of this interaction is the direct chelation of calcium
ion by the 3- and 4-hydroxyls of mannose.4
The analogy of fucose with mannose complexed in the
mannose binding protein was used as the starting point
to develop a model of the complex sLex–selectin. The
known solution NMR structures of sLex were docked
into the ligand binding site of selectins. Interestingly,
the calcium–fucose interaction in E- or P-selectin orients
the ligand so that the negatively charged group of the
sialic acid moiety is in a suitable position for an ionic
interaction with Lys 113.7 From previous binding stud-
ies, this carboxylate group of the sialic acid residue was
precisely designated as part of the binding domain of
In replacing the fucose ring in sLex by the galactose
sugar of the sulfatides in which the 4- and 6-hydroxy
groups can act as calcium chelators, an additional acidic
functionality at position 3 of this galactose ring has to
be introduced in order to mimic the carboxylate group
of sialic acid in sLex or the sulfate group in sulfatides.
The new structure should, in theory, interact with the
two critical binding features of the selectins, Lys 113
and the calcium ion. The acidic functionality insertion
in position 2 of the galactose ring should also be con-
sidered since the 4- and 6-hydroxyl groups are still
available for a chelation with the calcium ion.
sLex glycotope along with the galactose residue.11À15
A
schematic representation of these published binding
interactions is shown in Figure 2.
In an effort to develop antagonists to the binding of selec-
tins ligands that would be chemically and structurally
simpler than sLex, the structural requirements pre-
viously described for sLex binding to selectins were
modeled. Several publications16À18 have recently repor-
ted sLex mimics that retain diverse degrees of carbohy-
drate character. Among them, the monosaccharide 2
Finally, the presence of another conserved lysine (Lys
111) proximal to Lys 113 (ꢀ4–5 A) in the selectin bind-
ing cavity was shown to be as crucial as Lys 113 for the
binding of the sulfatides.7 It is consequently reasonable
to consider this residue as an additional binding feature
in the active site of the protein. To take advantage of
the chemical nature of this additional residue in the
binding cavity, the malonate functionality was con-
templated. Both acidic groups should interact with Lys
(TBC-265) (Fig. 3) containing
a carboxylic acid
attached to a mannose sugar via a biphenyl template
was proved to be as potent as sLex.19 Moreover, this
compound inhibited neutrophil migration in rat and
Figure 1. Sialyl Lewis X (1).
Figure 2. Assumed binding interactions in selectin–sLex model.

A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
1397
Figure 3. Biphenyl mannoses.
113 and Lys 111. Examples of malonate derived sLex
mimetics have been reported22 and have shown, in most
cases, slight potency improvements over sLex. How-
ever, no proof of the binding to the two lysines was
established.
between the two binding features of the molecules, two
carbon chain spacers were evaluated (n=2 and 4). The
protection scheme used to obtain the adequately pro-
tected 1-thio-b-galactopyranoside common intermediate
6 (Scheme 1) involves initial formation of the related
3,4-acetonide 525 followed by selective silylation in
position 6. The synthesis of the butylmalonyl 1-thio-b-d-
galactopyranoside intermediate 18 (Scheme 1) was per-
formed by alkylating the 2-position of 1-thio-galacto-
pyranoside 6, in presence of sodium hydride, with 1,4-
dibromobutane. Subsequent malonic displacement of
the remaining bromide functionality in 7 gave the desired
compound 8. In order to study the effect of a 3-blocked
position in this series of compounds, intermediate 8 was
then submitted to a last protection sequence. Deblocking
of the 3, 4 and 6 positions in the galactose sugar by acidic
treatment and protection of the 4- and 6-hydroxy groups
with benzaldehyde dimethylacetal liberated the 3-position
which became available for a benzoylation reaction to
give compound 18.
This paper describes the preparation of series of mal-
onate derivatives of type I and II (Fig. 5) with the
objective of identifying potent P-selectin antagonists.
Their SARs, evaluated in P-selectin in vitro models and
in the dermal reverse passive Arthus reaction in rats will
give insights into the means by which these ligands
interact with the protein and would allow validating the
previous hypothesis elaborated for their possible bind-
ing mode.
Chemistry
The preparation of the different galactosides is outlined
in Schemes 1–6. Dimethyl(methylthio)sulfonium triflate
promoted glycosidations were used to obtain the differ-
ent glycolipids as described by Fugedi and Garegg.23
The key thioglycosides were all prepared from ethyl 1-
thio-b-d-galactopyranoside24 employing the usual pro-
tection schemes used in classical carbohydrate chem-
istry. Most of the final compounds were tested as
disodium salts by treatment of the diacids with an
aqueous solution of sodium bicarbonate.
All attempts to apply the same synthetic scheme for the
preparation of the ethylmalonyl analogue 20 appeared
to be unsuccessful due to elimination of the alkylating
agent. Consequently, the goal was achieved by alkyla-
tion of intermediate 6 with t-butylbromoacetate to give
compound 9, followed by reduction of the ester group
and transformation of the resulting alcohol 10 to a
mesylate group. The malonic substitution, catalytically
assisted by sodium iodide, was then feasible and the
desired ethylmalonyl 1-thio-b-d-galactopyranoside 12 was
obtained. Again, as for the 4C analogue, similar protec-
tion-deprotection steps were performed and intermediate
20 was available for the next glycosidation reactions.
2-Substituted galactoconjugates
The preparation of malonate substituted galactolipids in
position 2 is described in Schemes 1, 3 and 5. In the aim
of excluding any ambiguity in regard to the distance
To produce non-benzoylated galactoconjugates in posi-
tion 3 such as compounds 33, 34 (Scheme 3), and 62
(Scheme 5), intermediates 8 (n=2) or 12 (n=4) were
required since removal of all protecting groups could be
done in one step. Hence, coupling of d-erythro-azido-
sphingosine26 28 with 8 or 12 under solvent modified
Fugedi and Garegg23 procedure gave galactocerebrosides
Figure 4. Sulfatides (3).
Figure 5. Types of malonate derivatives.

1398
A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
29a and b or 31a and b. The choice of solvent appeared
to be very peculiar, since silylation of the glycosyl
acceptor followed by intramolecular glycosidation by
the free 6-hydroxyl group (35)27 under dimethyl(methyl-
thio)sulfonium triflate conditions was an important side
reaction. Use of a nonpolar solvent such as toluene or
methylene chloride overcomes this problem. The a:b
anomers ratio using these solvents varied from 1:1 to
2:1. Each resulting azido-sphingosine anomers were
separately reduced and acylated as described by
Schmidt et al.28 prior to trifluoroacetic acid treatment
that removed all the protecting groups. Compound 62a
or b bearing a methoxycarbonyloctyl side-chain29 as
lipid aglycone were obtained the same way (Scheme 5).
The glycosidation reaction afforded in this case a a:b
ratio of approximately 1:1.
3-Benzoylated galactolipids 41, 43 (Scheme 3), and 60
(Scheme 5) were synthesized from glycosides 18 (n=4)
or 20 (n=2) in which the benzoyl group is already in
place. Glycosidation with azido-sphingosine26 followed by
separation of the anomers gave galactocerebrosides 36a
and b (n=4) and 38a and b (n=2) in an approximately 7:3
a:b ratio. Usual hydrogen sulfide reduction of the azido
functionality and acylation of the resulting amine with
palmitoyl chloride afforded intermediates 37a and b
(n=4) and 39a and b (n=2). Two subsequent acidic
treatments removed the 4,6-benzylidene protecting
Scheme 1. (a) CH3C(OCH3)2CH3, PTSA; (b) TBDMSCl, pyridine, DMAP; (c) NaH, Br(CH2)4Br, DMF; (d) NaCH(CO2tBu)2, THF, DMF;
(e) NaH, BrCH2CO2tBu, DMF; (f) LAH, THF; (g) MsCl, NEt3, CH2Cl2; (h) nBu4NF, THF; (i) 3 N HCl, THF; (j) PhCH(OCH3)2, MeCN, PTSA;
(k) BzCl, py, DMAP.
Scheme 2. (a) PMBCl, NaH, DMF; (b) AcOH, THF; (c) PhCH(OCH3)2, PTSA, MeCN; (d) Br-X-Br, NaH, DMF; (e) NaCH(CO2tBu)2, THF,
DMF.

A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
1399
group and the t-butyl esters to provide the desired
galactolipids 41a and b and 43a and b. The same
synthetic scheme was followed for the preparation of
analogues carrying Lemieux’s lipopohilic entity (60a
and 60b).
hydrolysis and the 4,6-benzylidene 23 was generated
after reaction with benzaldehyde dimethylacetal.
Sodium hydride treatment of 23 gave the corresponding
3-alkoxide which was alkylated by 1,4-dibromobutane
or a-a0-dibromo-p-xylene. As for the 2-substituted ana-
logues, the resulting bromides 25 or 27 were displaced
by the anion of di-t-butyl malonate to give the desired
intermediates 26 (X=(CH2)4) or 27 (X=CH2C6H4CH2).
3-Substituted galactoconjugates
Malonate substituted galactolipids in position 3 were
prepared as reported in Schemes 2, 4, and 6. The non-
participating paramethoxybenzyl ether protecting group
in position 2 of the key ethyl 1-thio-galactopyranoside
26 and 27 (Scheme 2) was necesary since both a and b
anomers were required for the study. This blocking
group was also advantageously stable to alkylation
conditions as well as acidic conditions. Hence, ethyl
3-Malonyl substituted galactocerebrosides 52 and 57
(Scheme 4) were both obtained after glycosidation of 26
and azido-sphingosine 28. The coupling reaction gave a
3:2 mixture of the two anomers 44a and b using dioxane
as solvent. After separation of the anomers and usual
reduction of the azido functionality followed by palmi-
tic acid acylation, galactocerebrosides 45a and b were
obtained. Removal of the paramethoxybenzyl protect-
ing group in position 2 was done with 2,3-dichloro-5,6-
dicyano-1,4-benzoquinone and the resulting alcohols
1-thio-b-d-galactopyranoside
5 was alkylated with
p-methoxybenzyl chloride to give compound 21. For-
mation of the triol 22 was performed by mild acidic
Scheme 3. (a) DMTST, 2,6-di-tBu-4-Me-pyridine, solvent, mol. sieves; (b) (1) H2S, pyridine, H2O; (2) CH3(CH2)14COCl, NaOAc, THF; (c) TFA
90%; (d) TFA 90%, CH2Cl2.

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A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
48a and b were then submitted to two successive acidic
hydrolysis to liberate the 4- and 6-hydroxyls and
recover the malonic acid functionality in 54a and b. The
same exact procedure was used to prepare the con-
strained galactocerebrosides 56a or b which contain a
methylbenzyl spacer. The key intermediate 27 was
condensed with azido-sphingosine in acetonitrile and
afforded a 1:1 mixture of a:b anomers 46a and b which
were easily separated by chromatography. Introduction
of the acylamino side chain followed by removal of the
benzylidene and the tert-butyl protecting groups gave
the desired compounds 56a or b.
lipid entity were obtained following a similar procedure
as described in Scheme 6.
Biology
In vitro binding assays
Extravasation of neutrophils into sites of inflammation
is dependent on the initial interaction with endothelial
P-selectin. This is supported by in vitro evidence of leu-
kocyte rolling on P-selectin in flow cells30 as well as the
in vivo anti-inflammatory activity of P-selectin blocking
antibodies.31 P-selectin is thus a novel target and an
inhibitor may provide anti-inflammatory activity. Three
assays have been put into place in order to determine
the ability of compounds to inhibit the binding of
P-selectin. Two employ a truncated soluble P-selectin
On the other hand, the synthesis of benzoylated galac-
tocerebrosides 52a and b was achieved by benzoylation
of alcohols 48a and b followed by acidic hydrolysis of
the resulting galactolipids 50a and b. Galactoconjugates
66a and b bearing the methoxycarbonyloctyl chain as
Scheme 4. (a) DMTST, 2,6-di-tBu-4-Me-pyridine, solvent, mol. sieves; (b) (1). H2S, pyridine, H2O; (2) CH3(CH2)14COCl, NaOAc, THF; (c) DDQ,
CH2Cl2, H2O; (d) TFA 90%, CH2Cl2, (e) TFA 90%; (f) BzCl, pyridine, DMAP; (g) MeONa.

A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
1401
protein containing the lectin, EGF and two complement
repeat domains fused to an immunoglobulin tail, termed
receptor globulin (Rg), as described.20,32 In a cell free
assay, the P-selectin Rg was allowed to bind to a 96-
well ELISA plate coated with sulfatides (Sigma). The
P-selectin was bound to HRP-conjugated goat anti-
human IgG and this immunocomplex was incubated on
the antigen-coated plate in the presence or absence of
the test compounds. The bound complexes were detec-
ted by addition of buffer containing substrate and
3,30,5,50-tetramethylbenzidine. Color development was
monitored and following the 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 data was
expressed as percent inhibition, from which an IC50 was
calculated and reported in Table 1 as cell free data.
Probes) for 20 min followed by resuspension in com-
plete PBS. In the assay, the P-selectin Rg coated wells
were preincubated with an inhibitor for 15 min prior to
addition of the labeled cells at 2Â105 cells per 0.2 mL
final volume. Following a 30 min binding period, the
unbound cells were removed by aspiration and the
bound cells were detected by a plate fluorimeter. The
data were expressed as percent inhibition from which an
IC50 value was determined and reported in Table 1 as
HL-60:P-selRg data.
A third assay was also developed that employed HL-60
cells binding to thrombin-activated platelets that
express P-selectin.33 The platelets were labeled by incu-
bation with calcein acetoxymethyl ester for 10 min at
37 ꢁC. Without washing, the platelets were counted and
the concentration adjusted to 1Â107 per mL. The plate-
lets were then activated with human thrombin, washed
and resuspended in buffer and counted. Cell concentra-
tions were adjusted to 2Â107/mL for the platelets and
4Â106 for HL-60 cells. The tested compounds were
incubated with labeled platelets for 10 min before addi-
tion of HL-60 cells. The samples were analyzed on a
flow cytometer by measuring the forward scatter and
fluorescent intensity of the mixed cell population. The
platelets appeared at lower forward scatter values and
The second assay measures labeled HL-60 cells bound
to immobilized P-selectin Rg. A 96-well plate was
coated with anti-human IgG Fc antibody, non-specific
binding sites were blocked with 1% non-fat dry milk,
and P-selectin Rg was bound by the antibody. HL-60
cells (3Â107 per mL of calcium and magnesium free
phosphate buffered saline (PBS, Gibco) were labeled by
taking up calcein acetoxymethyl ester (Molecular
Scheme 5. (a) DMTST, 2,6-di-tBu-4-Me-pyridine, solvent, mol. sieves; (b) TFA 90%, anisole; (c) nBu4NF, THF, AcOH.

1402
A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
Scheme 6.
were excluded. The percentage of HL-60 cells in fluor-
escent (containing bound platelets) and non-fluorescent
populations was determined and plotted as a function of
inhibitor concentration. The inhibition of binding was
compared to standards which were treated with vehicle
alone (0% inhibition) or incubated with EDTA (100%
inhibition). IC50 values were calculated and reported as
HL-60/platelet values in Tables 1–3.
injection of BSA containing 125I labeled BSA to induce
the inflammation reaction. Inhibitors of adhesion mol-
ecules were then administered intravenously and after
4 h, the rats were sacrificed. To assess changes in vas-
cular permeability, the 125I content of 15 mm punch
biopsies was determined by gamma scintillation spec-
troscopy. The radioactivity in the tissue compared to the
level of radioactive albumin in the blood at the time of
sacrifice allows to calculate the plasma protein exudation
into skin. The skin accumulation of neutrophils was
determined from the tissue content of the marker enzyme
myeloperoxidase (MPO). To measure the MPO content,
the method of Bradley et al.33,34 was used. The data were
reported as the percent inhibition of the endpoint for a
single dose of test compound (10 mg/kg, iv).
In vivo testing
The in vivo test is a reverse passive Arthus reaction in
rats33 in which the interaction of antigen–antibody
complexes and complement leads to a localized influx of
neutrophils. These neutrophils release a variety of med-
iators that are associated with tissue damage and vas-
cular permeability. E-selectin has been shown to be
involved in the inflammation resulting from an Arthus
reaction33 but the role of P-selectin has not been deter-
mined. However it has been suggested that P-selectin is
known to be involved in neutrophil dependent lung
injury and P-selectin deficient mice exhibit delayed
inflammatory reactions.33
Results and Discussion
Table 1 compares a representative example of the mal-
onate series (43a) with sLex, galactosyl ceramide and
sulfatides. The sLex epitope had very poor inhibition of
binding, as expected based on previous reports. Binding
to sLex may depend on the presence of a clustered sac-
charide arrangement. The natural glycosylated poly-
peptide ligand from which sLex was released was
presumed to create these multivalent saccharide patches
that avidly recognize the selectins. It is noteworthy,
however, that the commercial sulfatides did show
The localized inflammation produced during a reverse
passive Arthus reaction was evaluated by measuring the
endpoints of vascular permeability (VP) and neutrophil
infiltration (MPO). Briefly, the rats were administered
intradermal injection of anti-BSA followed by intravenous
Table 1. In vitro and in vivo (RPA reaction) results of a selected malonate representative 43a compared to commercial sulfatides (Aldrich),
galactosylceramide and sLex
In vitro
Cell free
IC50 (mM)
In vitro
HL-60:PselRg
IC50 (mM)
In vitro
HL-60/platelet
IC50 (mM)
In vivo RPA
%inhibition VP
In vivo RPA
%inhibition MPO
43a
sLex a
8.2
>100
>50
49
>100
not active
16
12
>100
41
96
Galactosyl ceramide
Commercial Sulfatides (Sigma)
0
39.5
0
10
0.64
aAs already mentioned, sLex affinity was shown to be ꢀ0.5–1.0 mM, and therefore has not been tested at higher concentration than 100 mM in these
assays.

A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
1403
Table 2. In vitro and in vivo (RPA reaction) results of various 2- and 3-malonate substituted galactocerebrosides as inhibitors of P-selectin
In vitro
In vivo
Compound
a/b
R1
R2
R3
HL-60/Platelet IC50 (mM)
%inhibition VP
%inhibition MPO
33a
33b
34a
41a
41b
43a
43b
52a
52b
54a
54b
56a
56b
57a
a
b
a
a
b
a
b
a
b
a
b
a
b
a
b
Bz
Bz
Bz
Bz
Bz
Bz
Bz
Bz
Bz
Bz
Bz
Bz
Bz
H
(CH2)4CH(CO2H)2
(CH2)4CH(CO2H)2
(CH2)2CH(CO2H)2
(CH2)4CH(CO2H)2
(CH2)4CH(CO2H)2
(CH2)2CH(CO2H)2
(CH2)2CH(CO2H)2
H
H
H
Bz
Bz
Bz
>80
32
>100
19
34
0
60
4
38
14
84
62
96
8.8
12
8.5
9.6
6
22.9
45
1.3
2.9
40.8
2
41
Bz
Bz
Bz
H
H
H
H
H
H
(CH2)4CH(CO2H)2
(CH2)4CH(CO2H)2
(CH2)4CH(CO2H)2
(CH2)4CH(CO2H)2
CH2C6H4CH2
CH2C6H4CH2
(CH2)4CH(CO2H)2
SO3Na
0
9.6
33
16
43
17
82
31
77
77
Sulfatides (Sigma)
H
39.5
10
Table 3. In vitro and in vivo (RPA reaction) results of various 2- and 3-malonate substituted galactoconjugates as inhibitors of P-selectin
In vitro
In vitro
In vivo
Compound
R2
R3
HL-60:PselRg IC50 (mM)
HL-60/platelet
IC50 (mM)
%inhibition
VP
%inhibition
MPO
60a
60b
62a
62b
66a
66b
(CH2)2CH(CO2H)2
(CH2)2CH(CO2H)2
(CH2)2CH(CO2H)2
(CH2)2CH(CO2H)2
Bz
Bz
H
H
>200
>160
172
>100
159
73
>127
>100
>150
>150
>145
>145
2
18
34
Bz
Bz
H
(CH2)4CH(CO2H)2
(CH2)4CH(CO2H)2
SO3Na
Sulfatides (Sigma)
16
39.5
10
activity in both in vitro and in vivo assays. Even though
sulfatides may coaggregate in patches on cell surfaces as
is known for glycosphingolipids, these results show that
the ‘clustering’ factor is not the only binding require-
ment for this type of compound. Galactosyl ceramide
was inactive in these assays, despite the presence of the
ceramide moiety. The potent inhibition by compound
43a suggests that the combined structural features of this
series are involved in the specific recognition of P-selectin.
for this type of compounds. Hence, although structu-
rally different, the malonate substituted galactocerebro-
sides are favorably compared to the sulfatides.
In the 2-malonate substituted series, a rapid analysis of
results concerning the two different spacers (33a vs 34a,
41a vs 43a and 41b vs 43b) shows that similar activity
was found and that chain length, in this range, does not
seem to matter. The 4C spacer easily satisfies the 10 A
distance requirement that is approximately measured
between Lys113 and the calcium ion in the binding
cavity and one can imagine that its high flexibility can
allow a slight folding in order to reach the two anchor
Table 2 reports the results of all analogues bearing the
sphingosine side-chain. As can be seen, several of the
compounds are active in in vitro assays. Moreover, a
number of these compounds showed activity when tes-
ted in vivo. By comparison with sulfatides as a refer-
ence, a number of the compounds showed superior in
vivo activity. Derivatives 33a and 34a are the two com-
pounds showing inactivity while analogues 54a, 54b and
57a exhibits approximately 10- to 20-fold decrease in in
vitro binding. However, in vivo activity, when com-
pared to sulfatides, seems to be as good or even better
Figure 6. Internal H-bond in the non-benzoylated 2-malonate series.

1404
A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
points. On the other hand, it can be supposed that the
2C chain fulfills the required distance in a fully extended
way. A marked effect of the benzoyl group in position 3
is apparent for this series of compounds (cf. compounds
41a vs 33a, 41b vs 33b and 34a vs 43a), improving their
potency by 5-fold. This suggests the proximity of a
lipophilic pocket in this region that provides a favorable
interaction. One can also postulate the formation of an
internal hydrogen bond between position 3 and 2 in the
non-benzoylated analogues (cf. Fig. 6) that would affect
the conformation of the sugar, its rigidity and conse-
quently its orientation in the cavity. Other substituents
at this position should be studied in order to get a clear
idea of the reason of this behavior.
33b) as indicated by both in vivo and in vitro biological
tests. In contrast, benzoylated derivatives bearing a
malonate side-chain in position 2 are equally active in
vitro compared to the 3-substituted malonate com-
pounds, but substantially more active in vivo (cf. com-
pounds 41a and 43a vs 52a and 52b and 43b vs 52b).
This comparison suggests that the benzoyl entity is tol-
erated in the cavity by both 2- and 3-malonate sub-
stituted derivatives. Benzoylation is also tolerated in
vivo in the 2-malonate series. However, this substitution
is a negative factor for in vivo activity in the 3-malonate
series. Globally, the benzoylated 2-malonate analogues
are the more potent as pointed in both in vitro and in
vivo assays.
The effect of the anomeric center is not as clear.
a-Configuration seems to slightly improve in vivo
activity in the 3-benzoyl series (cf. compounds 41a vs
41b and 43a vs 43b), while b-configuration appears bet-
ter in non-benzoylated compounds (33b vs 33a). These
results suggest that the position of the sugar in the
pocket, which is controlled by the ceramide side-chain
orientation, is not very critical in the case of benzoy-
lated analogues since similar in vitro results are found.
The flexibility of the ceramide moiety which may extend
out from the protein’s binding groove, could explain
this behavior. Nevertheless, in the non-benzoylated ser-
ies, a restricted sugar conformation due to an internal
H-bond would support the large difference seen in in
vitro activity between both anomers. The preference
for a b-configuration obviously involves a critical
binding orientation in the cavity for these compounds.
The biological results of derivatives bearing a methoxy-
carbonyloctyl aglycone chain29 are presented in Table 3.
Several key representatives shown in Table 2 were pre-
pared with this moiety in order to assess the importance
of the ceramide side-chain. The methoxycarbonyloctyl
side chain, which was shown to enhance activity of sLex
analogues1 was proved to dramatically reduced the
potency of this series of compounds. The only com-
pound (60b) tested in vivo when compared to the cor-
responding cerebroside (41b) tends to show a similar
behavior. The sphingosine aglycone thus plays an
important role in the binding process. The size of the
P-selectin pocket likely excludes any possibility of a
sphingosine binding interaction with the protein. This
large lipophilic entity is probably directed towards the
solvent in a fully extended all-trans configuration as
already known by X-ray crystallography or high-reso-
lution NMR analysis of related compounds. The direc-
tion and the tilt of this part of the molecule is
nevertheless unknown and might be important. Vesicles,
micelles or bilayers can be formed with this lipidic moi-
ety that would direct the polar sugar head in its pocket
in a suitable way to interact with the binding features.
Coaggregation in patches on cell surfaces could be a
natural mechanism for generating specific selectin
recognition in the case of the glycosphingolipids by
mimicking the multivalent interactions that occur with
the natural ligand.
In the 3-malonate substituted series, a reverse effect
was noticed concerning the presence of a benzoyl
group on the sugar. Despite similar in vitro results,
in vivo tests seem to indicate a deleterious effect of
the presence of a 2-benzoyl as shown for compounds
52a and 54a. In this case, metabolism or transport
issues are probably involved. Considering in vitro
assays only, a preference for one of the anomers is
not obvious in this series. However, in vivo testing
indicates that in the non-benzoylated series (54a vs
54b), the potency of the a-anomer is particularly
enhanced. Again, bioavailability to the target tissue
or metabolic instability of the b-anomer are impli-
cated. The introduction of a rigid spacer between the
acidic functionality and the carbohydrate ring in this
series proved to be beneficial as shown by the in
vitro results of 56a and 56b (by comparison with 54a
and 54b). As in sLex, which is a relatively rigid
molecule, selectin specificity gains are expected with
constrained analogues such as 54a and b which contain
a methylbenzyl template. However, in vivo activity of
these compounds, although very good, was not
enhanced by the same magnitude. Similar results were
found for both series which is not reflected in the in
vitro activity.
Conclusion
The selectins are important mediators of inflammatory
reactions. The discovery of a high-affinity selectin
antagonist can lead to potent anti-inflammatory agents
for the treatment of accute reactions. This study has
shown the high-affinity binding of malonate substituted
galactocerebrosides to P-selectin and their high poten-
tial as P-selectin antagonist. The 3-benzoylated 2-mal-
onate galactocerebrosides, much more potent than sialyl
Lewis X, have been identified as a lead in this new class
of compounds.
The data presented in this study suggests that this
new family of compounds acts by mimicking binding
to the sLex binding pocket. This study has identified
several key structural elements in this family of com-
pounds. The malonate group at position 2 is clearly
Position comparison for the malonate substituted side-
chain on the sugar must consider the presence of the
benzoyl group. The absence of benzoyl clearly favors
the 3-substituted analogues (54a vs 33a and 34a, 54b vs

A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
1405
favored as well as the presence of a substituent at
position 3. The sphingosine side-chain is required for
binding activity and the a-configuration is preferred,
at least in vivo. Preliminary studies with a more rigid
spacer as the methylbenzyl template indicate
enhancement of activity in vitro. The introduction of
rigidity in this class of molecules appears to be a
promising approach to more optimally mimic the
conformation of sLex and increase the binding affinity
to P-selectin.
isopropylidene-1-thio-b-d-galactopyranoside 5 (8.65 g,
32.7 mmol) in dry pyridine (125 mL) was treated at 0–
5 ꢁC with tert-butyldimethylsilyl chloride (5.92 g,
39.2 mmol) and the resulting mixture was stirred for 5 h.
Methanol (15 mL) was then added and the solution was
stirred for another 15 min. The solvent was then eva-
porated under vacuum and the residue was diluted with
ethyl acetate (500 mL) washed with cold 2 N hydro-
chloric acid, saturated sodium bicarbonate, brine and
dried over anhydrous magnesium sulfate. Evaporation
of the solvent under vacuum gave an oil which was
purified by chromatography on silica gel (9Â11 cm, 10–
20% ethyl acetate/toluene) and afforded the title
compound (12.4 g, 100%) as an oil. [a]2D2 +2.8ꢁ (c 1.0,
CHCl3). 1H NMR (400 MHz, CDCl3) d 0.08 (6H, s,
Si(CH3)2), 0.9 (9H, s,SitBu), 1.32 (3H, t, J=7.5 Hz,
-SCH2CH3) 1.35 and 1.53 (2Â3H, 2s, -C(CH3)2), 2.7
(2H, m, -SCH2CH3), 3.56 (1H, dd, J=10.2, 7.0 Hz, H-
2), 3.8–3.9 (3H, m, H-5 and H-6), 4.05 (1H, dd, J=7.0,
5.5 Hz, H-3), 4.24 (1H, d, J=10.2 Hz, H-1) 4.26 (1H,
dd, J=5.5 and 2.0 Hz, H-4).
Experimental
Material and methods
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
metling point apparatus and were not corrected. 1H
NMR spectra were measured on a Bruker AC200
(200 MHz) or a Bruker AMX400 (400 MHz) instru-
ments. Chemical shifts were reported in d units using the
solvent as internal standard. The abbreviations for the
multiplicity are the following: s=singlet, d=doublet,
t=triplet, q=quartet, m=multiplet. Infra-red spectras
were recorded on a Perkin-Elmer 781 and optical rota-
tions were measured on a Perkin-Elmer 241 apparatus.
Elemental analysis were performed at the Analytical
center of Bristol-Myers Squibb in Syracuse.
Ethyl 3,4-O-isopropylidene-6-O-tert-butyldimethylsilyl-2-
O-(4-bromobutyl)-1-thio-ꢀ-D-galactopyranoside 7. A stir-
red solution of ethyl 6-O-tert-butyldimethylsilyl-3,4-O-
isopropylidene-1-thio-b-d-galactopyranoside 6 (0.89 g,
2.35 mmol) in N,N-dimethylformamide (20 mL) was
treated with sodium hydride (60% in mineral oil, 0.90 g,
22.5 mmol) at 0 ꢁC. The mixture was stirred at room
temperature for 10 min, then 1,4-dibromobutane (6 mL,
50.2 mmol) was added. The mixture was stirred for ꢀ30
min at 22 ꢁC, then diluted with ethyl ether and washed
with saturated ammonium chloride, water and brine.
The residue was purified by silica gel chromatography
(5–8% ethyl acetate/hexane) to give the title compound
Key intermediates
1
Ethyl 3,4-O-isopropylidene-1-thio-ꢀ-D-galactopyranoside
5.25 A mixture of ethyl 1-thio-b-d-galactopyranoside
424 (24.86 g, 0.111 mol) and 2,2-dimethoxypropane
(500 mL) was treated with p-toluenesulfonic acid
(0.625 g) and stirred at 22 ꢁC for 24 h. Water (80 mL)
was added and after 15 min the reaction mixture was
cooled in an ice water bath and stirred for another 30
min. Then triethylamine (5 mL) was added and the
mixture was stirred for 20 min. The solvent was evapo-
rated under vacuum and the residue was purified by
silica gel chromatography (9Â12 cm, 50–70% ethyl
acetate/toluene) to give the title material (25.50 g, 87%)
as a white solid. Recrystallization from ethyl acetate
and hexane gave white prisms (mp 90–93 ꢁC). [a]D22
(0.950 g, 79%) as an oil. H NMR (200 MHz, CDCl3)
d 0.07 (6H, s, -Si(CH3)2), 0.89 (9H, s, -SitBu), 1.29
(3H, t, J=7.5 Hz, -CH2–CH3), 1.34 and 1.52 (6H, 2s,
-(CH3)2C-), 1.67–1.80 and 1.92–2.06 (4H, 2m, -(CH2)2-),
2.62–2.82 (2H, m, -SCH2-), 3.26 (1H, dd, J=9.9, 6.7 Hz,
H-2), 3.47 (2H, t, J=6.7 Hz, -CH2Br), 3.62–3.83 (4H,
m, H-6 and -OCH2-), 3.86 (1H, d, J=1.9 Hz, H-50), 4.07
(1H, dd, J=5.5, 6.7 Hz, H-3), 4.22 (1H, dd, J=5.5,
1.9 Hz, H-4), 4.32 (1H, d, J=9.9 Hz, H-1). IR (CH2Cl2)
ꢀmax (cmÀ1) 2940, 2920, 2840 (C–H).
Ethyl 3,4-O-isopropylidene-6-O-tert-butyldimethylsilyl-2-
O-(5,5-di-tert-butyloxycarbonylpentyl)-1-thio-ꢀ-D-galac-
topyranoside 8. A stirred solution of di-tert-butylmalo-
nate (3.0 mL, 13.4 mmol) in tetrahydrofuran (20 mL)
was treated with a solution of potassium tert-butoxide
in tert-butanol (1.0 M, 15.0 mL, 15 mmol) at 0 ꢁC. The
mixture was stirred for ꢀ30 min. A solution of ethyl 3,4-
O-isopropylidene-6-O-tert-butyldimethylsilyl-2-O-bromo-
butyl-1-thio-b-d-galactopyranoside 7 (4.00 g, 7.79 mmol)
in N,N-dimethylformamide (20 mL) was then added to
this mixture and the ice-cooled bath was removed. The
mixture was heated to ꢀ50–60 ꢁC for ꢀ2 h, then diluted
with ethyl acetate. A saturated solution of ammonium
chloride was added and the two phases were separated.
The organic layer was then washed with water and
brine, dried over anhydrous magnesium sulfate, filtered
and concentrated. The residue was purified by silica gel
+20.8ꢁ (c 2.8, CHCl3). H NMR (400 MHz, CDCl3) d
1
1.33 (3H, t, J=7.3 Hz, -SCH2CH3), 1.36 and 1.52
(2Â3H, 2s, -C(CH3)2), 2.2 and 2.5 (broad, OH), 2.75
(2H, m, -SCH2CH3), 3.57 (1H, dd, J=10.2, 7.0 Hz, H-
2), 3.81 (1H, dd, J=11.5, 4.0 Hz, H-6), 3.89 (1H, m,
H-5), 3.98 (1H, dd, J=11.5, 7.2 Hz, H-6), 4.09 (1H,
dd, J=7.0, 5.6 Hz, H-3), 4.21 (1H, dd, J=5.6, 2.2 Hz,
H-4), 4.27 (1H, d, J=10.2 Hz, H-1). IR (KBr) ꢀmax
(cmÀ1) 3200 (br, OH). Anal. calcd for C11H20O5S: C,
49.98; H, 7.63; S, 12.13. Found: C, 49.89; H, 7.49; S,
12.35.
Ethyl 6-O-tert-butyldimethylsilyl-3,4-O-isopropylidene-1-
thio-ꢀ-D-galactopyranoside 6. A solution of ethyl 3,4-O-

1406
A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
chromatography (5–10% ethyl acetate/hexane) and
1
-Si(CH3)2), 0.90 (9H, s, -SitBu), 1.31 (3H, t, J=7.4 Hz,
-CH3), 1.35 and 1.54 (6H, 2s, -C(CH3)2), 2.66–2.82 (2H,
m, -SCH2-), 2.92 (1H, t, J=6.5 Hz, -OH), 3.55 (1H, dd,
J=10.0, 7.0 Hz, H-2), 3.63–3.74 (2H, m, -CH2OH), 3.77
(1H, m, H-5), 3.82–3.90 (4H, m, -OCH2- and H-6), 4.12
(1H, dd, J=6.8, 5.5 Hz, H-3), 4.26 (1H, dd, J=5.5,
2.1 Hz, H-4), 4.33 (1H, d, J=10.0 Hz, H-1). IR
(CH2Cl2) ꢀmax (cmÀ1) 3490 (OH), 3050, 2990, 2935,
2860 (C-H). Anal. calcd for C19H38O6SSi: C, 53.99; H,
9.06; S, 7.59. Found: C, 53.82; H, 8.91; S, 7.65.
afforded the title compound (5.2 g, 100%) as an oil. H
NMR (400 MHz, CDCl3) d 0.08 (6H, s, -Si(CH3)2), 0.90
(9H, s, -SitBu), 1.29 (3H, t, J=7.4 Hz, -CH2–CH3), 1.34
(3H, s, -C(CH3)2-), 1.36–1.43 (2H, m, -CH2-), 1.47
(18H, s, 2 Â-OtBu), 1.52 (3H, s, -C(CH3)2-), 1.60–1.67
and 1.83 (4H, 2m, -(CH2)2-), 2.64–2.80 (2H, m, -SCH2-),
3.13 (1H, t, -CH(CO2tBu)2), 3.28 (1H, dd, J=9.8,
6.7 Hz, H-2), 3.65 (1H, dt, J=9.3, 6.8 Hz, -OCH2-),
3.72–3.79 and 3.82–3.89 (4H, 2m, -OCH2-, H-5 and H-
6), 4.09 (1H, br dd, H-3), 4.22 (1H, dd, J=5.5, 2.0 Hz,
H-4), 4.33 (1H, d, J=9.8 Hz, H-1). IR (CH2Cl2) ꢀmax
(cmÀ1) 2930, 2850 (C–H), 1740, 1725 (C¼O).
Ethyl 3,4-O-isopropylidene-6-O-tert-butyldimethylsilyl-2-
O-methanesulfonyloxyethyl-1-thio-ꢀ-D-galactopyranoside
11. Triethylamine (99 mL, 0.711 mmol) followed by
methanesulfonyl chloride (44 mL, 0.569 mmol) were
added to a stirred solution of ethyl 3,4-O-isopropylidene-
6-O-tert-butyldimethylsilyl-2-O-hydroxyethyl-1-thio-b-d-
galactopyranoside 10 (0.200 g, 0.474 mmol) in methyl-
ene chloride (2 mL) at 0 ꢁC. The mixture was stirred for
30 min then water (ꢀ2 mL) was added. The aqueous
layer was extracted with methylene chloride (3Â3 mL).
The combined organic layers were dried over anhydrous
magnesium sulfate, filtered and concentrated. The
residue was purified by silica gel column chromato-
graphy (30% ethyl acetate/hexane) and afforded the
title compound (0.229 g, 97%) as an oil. 1H NMR
(200 MHz, CDCl3) d 0.07 (6H, s, -Si(CH3)2-), 0.89 (9H,
s, SitBu), 1.28 (3H, t, J=5.2 Hz, -CH3), 1.33 and 1.52
(6H, 2s, -C(CH3)2), 2.72 (2H, m, -SCH2-), 3.09 (3H, s,
-SO2CH3), 3.31 (1H, dd, J=9.9, 6.6 Hz, H-2), 3.74–
4.17 (7H, m, H-6, H-5, -OCH2- and -CH2OMs), 4.24
(1H, dd, J=5.4, 1.8 Hz, H-4), 4.35–4.41 (2H, m, H-1 and
H-3).
Ethyl 6-O-t-butyldimethylsilyl-3,4-O-isopropylidene-2-O-
tert-butyloxycarbonylmethyl-1-thio-ꢀ-D-galactopyrano-
side 9. A solution of ethyl 6-O-tert-butyldimethylsilyl-
3,4-O-isopropylidene-1-thio-b-d-galactopyranoside
6
(6.7 g, 17.7 mmol) in dimethylformamide (120 mL) was
added to sodium hydride (4.25 g, 50% suspension in oil,
previously washed with hexane, 88 mmol) under argon.
The resulting suspension was stirred at 22 ꢁC for 0.75 h.
tert-Butyl bromoacetate (16.5 mL, 102 mmol) was added
dropwise to this mixture, which was stirred for an
additional h. The mixture was then diluted with cold
1 M aqueous sodium bicarbonate (150 mL) and ethyl
acetate (250 mL). The organic phase was washed with
water (4Â150 mL), 1 M aqueous sodium bicarbonate
(150 mL) and brine (150 mL), dried over anhydrous
magnesium sulfate, filtered and concentrated. The resi-
due was purified by silica gel column chromatography
(300 g, 2–10% ethyl acetate/hexane) to give the title
compound (8.03 g, 92%) as a yellow oil. 1H NMR
(400MHz, CDCl3) d 0.07 (6H, s, -Si(CH3)2), 0.90 (9H, s,
-SitBu), 1.30 (3H, t, J=7.4 Hz, -CH3), 1.33 and 1.50 (6H,
2s, -C(CH3)2), 1.48 (9H, s,-OtBu), 2.67–2.82 (2H, m,
-SCH2-), 3.48 (1H, dd, J=9.7, 6.0 Hz, H-2), 3.75 (1H, td,
J=6.6, 1.3 Hz, H-5), 3.82–3.89 (2H, m, H-6), 4.21 (1H, d,
J=16.2, -OCH2CO-), 4.19–4.25 (2H, m, H-3 and H-4),
4.33 (1H, d, J=16.2 Hz, -OCH2CO-), 4.42 (1H, d,
J=9.7 Hz, H-1). IR (CH2Cl2) ꢀmax (cmÀ1) 3055, 2990,
2835 (C–H), 1745 (C¼O). Anal. calcd for C23H44O7SSi: C,
56.06; H, 9.00; S, 6.51. Found: C, 56.20; H, 8.79; S, 6.67.
Ethyl 3,4-O-isopropylidene-6-O-tert-butyldimethylsilyl-2-
O-(3,3-di-tert-butyloxycarbonylpropyl)-1-thio-ꢀ-D-galacto-
pyranoside 12. tert-Butyl malonate (225 mL, 1.00 mmol)
was added to a suspension of sodium hydride (36 mg,
80% suspension in oil, 1.20 mmol) in dimethylforma-
mide (2 mL) at 0 ꢁC. This mixture was stirred at 22 ꢁC for
0.75 h, then a solution of ethyl 3,4-O-isopropylidene-6-O-
tert-butyldimethylsilyl-2-O-methanesulfonyloxy-ethyl-1-
thio-b-d-galactopyranoside 11 (0.102 g, 0.204 mmol) in
tetrahydrofuran (4 mL) was added in dropwise, fol-
lowed by solid potassium iodide (ꢀ7 mg, 0.04 mmol).
The resulting mixture was stirred at 65 ꢁC for 12 h, then
diluted with ethyl acetate (ꢀ10 mL) and quenched with
saturated ammonium chloride. The organic layers were
washed with water (10 mL), dried over anhydrous mag-
nesium sulfate, filtered and concentrated. The residue
was purified by silica gel column chromatography (5–
10% ethyl acetate/hexane) and afforded the title mat-
erial (0.330 g, contaminated with tert-butyl malonate).
An aliquot of pure material was purified and obtained
Ethyl 6-O-tert-butyldimethylsilyl-3,4-O-isopropylidene-2-
O-hydroxyethyl-1-thio-ꢀ-D-galactopyranoside 10.
A
solution of ethyl 6-O-tert-butyldimethylsilyl-3,4-O-iso-
propylidene-2-O-tert-butyloxycarbonylmethyl-1-thio-b-d-
galactopyranoside
9
(4.0 g, 8.1 mmol) in tetra-
hydrofuran (40 mL) was treated with lithium aluminium
hydride (0.31 g, 8.1 mmol) which was added by portions
at 5 ꢁC. The reaction mixture was stirred for 1 h. The
same quantity of lithium aluminium hydride is added
again and the mixture was stirred for another h. The
mixture was then diluted with ethyl ether, cooled down
to À15 ꢁC and treated with a 3 N aqueous solution of
sodium hydroxide. This was stirred for 0.75 h, then the
organic layer was separated and the residual gum was
washed with ethyl ether (3Â100 mL). The combined
organic layers were then dried over anhydrous magne-
sium sulfate, filtered and concentrated. The residue
(2.73 g, 80%) without any purification was used for the
next reaction. 1H NMR (400 MHz, CDCl3) d 0.08 (6H, s,
1
as an oil. H NMR (200 MHz, CDCl3) d 0.06 (6H, s,
-Si(CH3)2), 0.88 (9H, s, -SitBu), 1.28 (3H, t, J=3.2 Hz,
-CH3), 1.31 and 1.49 (6H, 2s, -C(CH3)2), 1.45 (18H, br
s, 2Â-OtBu), 2.08 (2H, m, -CH2–CH(CO2tBu)2), 2.70
(2H, m, -SCH2-), 3.24 (1H, dd, J=9.8, 6.6 Hz, H-2),
3.46 (1H, t, J=7.4 Hz, -CH(CO2tBu)2), 3.63–3.85
(5H, m, H-5, H-6, -OCH2-), 4.08 (1H, br t, H-3), 4.21
(1H, dd, J=5.5, 1.8 Hz, H-4), 4.30 (1H, d, J=9.8 Hz,
H-1).

A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
1407
Ethyl 3,4-O-isopropylidene-2-O-(5,5-di-tert-butyloxycar-
bonylpentyl)-1-thio-ꢀ-D-galactopyranoside 13. A solution
of ethyl 3,4-O-isopropylidene-6-O-tert-butyldimethylsi-
lyl-2-O-(5,5-di-tert-butyloxycarbonylpentyl)-1-thio-b-d-
galactopyranoside 8 (5.0 g, 7.7 mmol) in tetrahydro-
furan (30 mL) was treated with tetra-butylammonium
fluoride (3.09 mL, 1 M solution in tetrahydrofuran,
3.09 mmol) at 0 ꢁC. The mixture was stirred at 22 ꢁC for
ꢀ2 h, then diluted with ethyl acetate (ꢀ60 mL) and
washed with water (ꢀ60 mL). The organic layer was
dried over anhydrous magnesium sulfate, filtered, and
concentrated. The residue was purified by silica gel col-
umn chromatography and gave the title compound
(3.8 g, 86%) as an oil. 1H NMR (400 MHz, DMSO-d6) d
1.19 (3H, t, J=7.4 Hz, -CH2-CH3), 1.24–1.42 (23H, m,
2Â-OtBu, -CH2-, -C(CH3)2-), 1.26 (3H, s, -C(CH3)2-),
1.47 (2H, ap q, -CH2-), 1.67 (2H, ap q, -CH2-), 2.63 (2H,
m, -SCH2-), 3.09 (1H, dd, J=9.8, 6.8 Hz, H-2), 3.14 (1H,
t, J=7.5 Hz, -CH(CO2tBu)2), 3.50–3.55 (3H, m, H-6 and
-OCH2-), 3.62 (1H, dt, J=9.5, 6.2 Hz, -OCH2-), 3.74 (1H,
td, J=6.3, 1.8 Hz, H-5), 4.05 (1H, br dd, H-3), 4.15 (1H,
dd, J=5.5, 1.9 Hz, H-4), 4.38 (1H, d, J=9.8 Hz, H-1),
J=7.5 Hz, -CH(CO2tBu)2), 3.43–3.48 (2H, m, H-2 and
H-5), 3.68 (1H, dd, J=8.4, 3.6 Hz, H-3), 3.70–3.77 (1H,
m, -OCH2-), 3.84 (1H, dt, J=8.8, 6.4 Hz, -OCH2-), 4.03
(1H, dd, J=12.4, 1.7 Hz, H-6), 4.26 (1H, d, J=3.5 Hz,
H-4), 4.34 (1H, dd, J=12.4, 1.3 Hz, H-6), 4.36 (1H, d,
J=9.5 Hz, H-1), 5.55 (1H, s, -OCHO-), 7.37–7.41 and
7.51–7.53 (5H, 2 sets of m, aromatic H).
Ethyl 3-O-benzoyl-4,6-O-benzylidene-2-O-(5,5-di-tert-butyl-
oxycarbonylpentyl)-1-thio-ꢀ-D-galactopyranoside 18.
A
stirred solution of ethyl 4,6-O-benzylidene-2-O-(5,5-di-
tert-butyloxycarbonylpentyl)-1-thio-b-d-galactopyrano-
side 17 (2.1 g, 3.56 mmol) in methylene chloride (20 mL)
and pyridine (20 mL) was treated with benzoyl chloride
(530 mL, 4.58 mmol) and 4-dimethylaminopyridine
(ꢀ10 mg) at 22 ꢁC. The mixture was stirred for 4 h then
diluted with methylene chloride (40 mL) and washed
with aqueous 1 M sodium bicarbonate (ꢀ40 mL) and
brine (ꢀ40 mL). The organic layer was dried over
anhydrous magnesium sulfate, filtered and con-
centrated. The residue was purified by silica gel column
chromatography (10–30% ethyl acetate/hexane) and
1
4.78 (1H, t, J=5.6 Hz, -OH). IR (CH2Cl2) ꢀmax (cmÀ1
)
3700–3100 (OH), 2980, 2930, 2860 (C–H), 1745, 1725
(C¼O).
gave the title material (2.1 g, 85%) as an oil. H NMR
(400 MHz, CDCl3) d 1.18–1.30 (2H, m, -CH2-), 1.34
(3H, t, J=7.5 Hz, -CH2–CH3), 1.42 and 1.43 (18H, 2s,
2Â -OtBu), 1.48–1.54 (2H, m, -CH2-), 1.60–1.73 (2H, m,
-CH2-), 2.73–2.90 (2H, m, -SCH2-), 2.95 (1H, t,
J=7.6 Hz, -CH(CO2tBu)2), 5.58 (1H, br s, H-5), 3.64
(1H, dt, J=8.9, 6.6 Hz, -OCH2-), 3.81 (1H, dt, J=8.9,
6.7 Hz, -OCH2-), 3.85 (1H, t, J=9.6 Hz, H-2), 4.03 (1H,
dd, J=12.4, 1.6 Hz, H-6), 4.36 (1H, dd, J=12.4, 1.3 Hz,
H-6), 4.50 (1H, d, J=3.5 Hz, H-4), 4.51 (1H, d,
J=9.5 Hz, H-1), 5.14 (1H, dd, J=9.6, 3.5 Hz, H-3), 5.50
(1H, s, -OCHO-), 7.35–7.39, 7.44–7.50, 7.55–7.60 and
8.07–8.09 (10H, 4 sets of m, aromatic H). IR (CH2Cl2)
Ethyl 2-O-(5,5-di-tert-butyloxycarbonylpentyl)-1-thio-ꢀ-
D-galactopyranoside 15. A solution of ethyl 3,4-O-iso-
propylidene-2-O-(5,5-di-tert-butyloxycarbonylpentyl)-1-
thio-b-d-galactopyranoside 13 (3.5 g, 6.1 mmol) in 80%
acetic acid (20 mL) was stirred for ꢀ72 h at 22 ꢁC. The
solvents were evaporated and the residue was purified
by silica gel chromatography (0–5% methanol/ethyl
acetate) to give the title compound (3.0 g, 99%) as an
oil. 1H NMR (400 MHz, DMSO-d6) d 1.19 (3H, t,
J=7.4 Hz, -CH3), 1.36–1.24 (2H, m, -CH2-), 1.40 (18H,
s, 2Â -OtBu), 1.56–1.45 (2H, m, -CH2-), 1.67 (2H, ap q,
J=7.7 Hz, -CH2-), 2.62 (2H, m, -SCH2-), 3.67–3.62,
3.57–3.52, 3.50–3.42, 3.40–3.29 and 3.15–3.11 (9H, 5 sets
of m, H-2, H-3, H-4, H-5, H-6, -OCH2- and -CH-
(CO2tBu)2), 4.24 (1H, d, J=9.6 Hz, H-1), 4.48 (1H, d,
J=4.6 Hz, -OH), 4.54 (1H, t, J=5.6 Hz, -OH-6), 4.78
(1H, d, J=6.6 Hz, -OH). IR (nujol) ꢀmax (cmÀ1): 3700–
3000 (br, OH), 2910 (br, C–H), 1740, 1710 (C¼O).
ꢀ
max (cmÀ1) 2970, 2920, 2860 (C–H), 1740, 1720 (C¼O).
Ethyl 3,4-O-isopropylidene-2-O-(3,3-di-tert-butyloxycar-
bonylpropyl)-1-thio-ꢀ-D-galactopyranoside 14. Ethyl 3,4-
O-isopropylidene-6-O-tert-butyldimethylsilyl-2-O-(3,3-di-
tert-butyloxycarbonylpropyl)-1-thio-b-d-galactopyrano-
side 12 (0.330 g, contaminated with tert-butyl malonate)
was reacted by the procedure used to synthesized com-
pound 13 and gave the title compound (0.069 g, 67% for
2 steps) as an oil. [a]2D2 –1.2ꢁ (c 1.2, CHCl3). H NMR
1
Ethyl 4,6-O-benzylidene-2-O-(5,5-di-tert-butyloxycarbo-
nylpentyl)-1-thio-ꢀ-D-galactopyranoside 17. A solution
of ethyl 2-O-(5,5-di-tert-butyloxycarbonylpentyl)-1-
thio-b-d-galactopyranoside 15 (3.0 g, 6.05 mmol) in
acetonitrile (50 mL) was treated with benzaldehyde
dimethylacetal (4.5 mL, 30.3 mmol) and p-toluene-
sulfonic acid (150 mg). The mixture was stirred at 22 ꢁC
for 1 h, then was diluted with ethyl acetate (50 mL) and
washed with aqueous 1 M sodium bicarbonate
(ꢀ50 mL) and brine (ꢀ50 mL). The organic layer was
dried over anhydrous magnesium sulfate, filtered and
concentrated. The residue was purified by silica gel col-
umn chromatography (30–50% ethyl acetate/hexane)
and afforded the title material (2.1 g, 59%) as an oil.
1H NMR (400 MHz, CDCl3) d 1.34 (3H, t, J=7.5 Hz,
-CH2-CH3), 1.37–1.49 (2H, m, -CH2-), 1.46 (18H, 2s,
2Â-OtBu), 1.62–1.68 and 1.78–1.87 (4H, 2m, -(CH2)2-),
2.71–2.86 (3H, m, -SCH2- and -OH), 3.13 (1H, t,
(400 MHz, CDCl3) d 1.31 (3H, t, J=7.7 Hz, -CH3), 1.34
and 1.51 (6H, 2s, -C(CH3)2), 1.47 (18H, s, 2Â -OtBu),
2.05–2.12 (3H, m, -CH2CH(CO2tBu)2 and -OH), 2.74
(2H, m, -SCH2-), 3.28 (1H, dd, J=9.5 and 6.2 Hz, H-2),
3.47 (1H, t, J=7.4 Hz, -CH(CO2tBu)2), 3.68 (1H, dt,
J=9.6, 6.3 Hz, -OCH2-), 3.76–3.84 and 3.92–3.99 (4H, 2
sets of m, -OCH2-, H-6 and H-5), 4.14 (1H, br dd, H-3),
4.18 (1H, dd, J=5.8, 1.7 Hz, H-4), 4.35 (1H, d,
J=9.5 Hz, H-1). IR (film) ꢀmax (cmÀ1) 3700–3020 (OH),
2970, 2920 (C-H), 1720 (C¼O). Anal. calcd for
C24H42O9S: C, 56.90; H, 8.36; S, 6.33. Found: C, 56.83;
H, 8.14; S, 6.59.
Ethyl 2-O-(3,3-di-tert-butyloxycarbonylpropyl)-1-thio-ꢀ-
D-galactopyranoside 16. A solution of ethyl 3,4-O-iso-
propylidene-2-O-(3,3-di-tert-butyloxycarbonylpropyl)-1-
thio-b-d-galactopyranoside 14 (0.885 g, 1.75 mmol) in
tetrahydrofuran (20 mL) was treated with aqueous

1408
A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
hydrochloric acid (2 N, 5 mL). The mixture was stirred
at 22 ꢁC for 12 h and then diluted with ethyl acetate
(25 mL). Solid sodium bicarbonate was added at 0 ꢁC,
followed by anhydrous magnesium sulfate. The mixture
was filtered and the solvents were evaporated. The resi-
due was purified by silica gel column chromatography
(30% ethyl acetate/hexane to pure ethyl acetate) to give
the title compound (0.758 g, 93%) as an oil. [a]D22
(cmÀ1) 3035, 3010, 2970, 2920, 2860 (C–H), 1720
(C¼O). Anal. calcd for C35H46O10S: C, 63.81; H, 7.04;
S, 4.87. Found: C, 63.94; H, 6.89; S, 5.41.
Ethyl 6-O-tert-butyldimethylsilyl-3,4-O-isopropylidene-2-
O-p-methoxybenzyl-1-thio-ꢀ-D-galactopyranoside 21. A
solution of ethyl 6-O-tert-butyl-dimethylsilyl-3,4-O-iso-
propylidene-1-thio-b-d-galactopyranoside
6
(12.4 g,
À25.9ꢁ (c 1.1, CHCl3). H NMR (400 MHz, CDCl3) d
32.7 mmol) in N,N-dimethylformamide (45 mL) was
added to a suspension of sodium hydride (1.35 g of 80%
in mineral oil, 45.0 mmol) in N,N-dimethylformamide
(90 mL) and the resulting mixture was stirred at 22 ꢁC
for 2 h. The reaction mixture was then cooled to 0–5 ꢁC,
treated dropwise with p-methoxybenzyl chloride
(8.1 mL, 59.8 mmol) and stirred at 22 ꢁC for 2 h. The
reaction mixture was cooled again in an ice bath and
treated dropwise with water (20 mL). The reaction mix-
ture was then diluted with water (300 mL) and extracted
with ether (3Â300 mL). The combined extracts were
washed with brine, dried over anhydrous magnesium
sulfate and evaporated. The residual oil was purified on
silica gel chromatography (9Â12 cm, 0–10% ethyl ace-
tate/toluene) and gave the title compound (10.5 g, 64%)
1
1.32 (3H, t, J=7.5 Hz, -CH3), 1.47 and 1.48 (18H, 2s,
2Â -OtBu), 1.98–2.06 (1H, m, -OH), 2.15–2.28 (2H, m,
-CH2CH(CO2tBu)2), 2.76 (2H, m, -SCH2-), 2.80 (1H, br
s, -OH), 3.34 (1H, br dd, H-2), 3.45–3.54 (3H, m, H-5,
-CH(CO2tBu)2 and -OCH2-), 3.59 (1H, td, J=8.8 and
3.3 Hz, -OCH2-), 3.82 (1H, ddd, J=11.8, 8.4 and 4.3 Hz,
H-6), 3.97 (1H, ddd, J=11.8, 6.6 and 4.4 Hz, H-6), 4.04
(1H, d, J=1.2 Hz, -OH-4), 4.17 (1H, dd, J=8.9 and
4.4 Hz, H-3), 4.31 (1H, d, J=9.7 Hz, H-1), 4.38 (1H, br
d, J=3.2 Hz, H-4). IR (film) ꢀmax (cmÀ1) 3700–3020
(OH), 2970, 2920 (C–H), 1720 (C¼O). Anal. calcd for
C21H38O9S.0.3H2O: C, 53.44; H, 8.24; S, 6.79. Found:
C, 53.39; H, 7.95; S, 6.76.
1
Ethyl
2-O-(3,3-di-tert-butyloxycarbonylpropyl)-4,6-O-
as an oil. H NMR (200 MHz, CDCl3) d 0.06 (6H, s,
benzylidene-1-thio-ꢀ-D-galactopyranoside 19. Ethyl 2-O-
(3,3-di-tert-butyloxycarbonylpropyl)-1-thio-b-d-galac-
topyranoside 16 (0.724 g, 1.55 mmol) in acetonitrile
(20 mL) was reacted by the procedure used to synthesize
compound 17 and afforded the title material (0.77 g,
SiCH3), 0.88 (9H, s, -SitBu), 1.29 (3H, t, J=7.4 Hz,
-SCH2CH3), 1.34 and 1.44 (2Â3H, 2s, -C(CH3)2), 2.7
(2H, m, -SCH2CH3), 3.41 (1H, dd, J=6.1, 9.9 Hz, H-2),
3.79 (3H, s, -OCH3), 3.7–3.9 (3H, m, H-5 and H-6),
4.15–4.25 (2H, m, H-3 and H-4 overlapping), 4.39 (1H,
d, J=9.9 Hz, H-1), 4.70 (1H, d, J=11.0 Hz, -OCH2Ar),
4.74 (1H, d, J=11.0 Hz, -OCH2Ar), 6.86 (2H, d,
J=8.7 Hz, H-3 of 4-methoxybenzyl) and 7.35 (2H, d,
J=8.7 Hz, H-2 of 4-methoxybenzyl). IR (NaCl, film)
ꢀmax (cmÀ1) 1612 (aromatic) and 1516.
22
89%) as an oil. [a]D : À32.8ꢁ (c 1.2, CHCl3). H NMR
(400 MHz, CDCl3) d 1.34 (3H, t, J=7.5 Hz, -CH3), 1.45
and 1.46 (18H, 2s, 2Â-OtBu), 2.02–2.21 (2H, m, -CH2–
CH(CO2tBu)2), 2.79 (2H, m, -SCH2-), 3.44 (1H, br s, H-
5), 3.46–3.51 (2H, m, -CH(CO2tBu)2 and H-2), 3.55
(1H, J=6.4 Hz, -OH), 3.64–3.69 and 4.05–4.08 (3H, 2
sets of m, H-3 and -OCH2-), 4.03 (1H, dd, J=12.7,
1.7 Hz, H-6), 4.26 (1H, d, J=3.5 Hz, H-4), 4.32 (1H, d,
J=9.5 Hz, H-1), 4.32 (1H, d, J=12.7 Hz, H-6), 5.55
(1H, s, -OCHO-), 7.35–7.39 and 7.50–7.53 (5H, 2 sets of
m, -C6H5). IR (film) ꢀmax (cmÀ1) 3600–3200 (OH), 2970,
2920, 2880 (C–H), 1735, 1720 (C¼O). Anal. calcd for
C28H42O9S: C, 60.63; H, 7.63; S, 5.78. Found: C, 60.74;
H, 7.46; S, 6.23.
1
Ethyl 2-O-p-methoxybenzyl-1-thio-ꢀ-D-galactopyrano-
side 22. A solution of ethyl 6-O-tert-butyldimethylsilyl-
3,4-O-isopropylidene-2-O-p-methoxybenzyl-1-thio-b-d-
galactopyranoside 21 (10.50 g, 21.05 mmol) in 80%
aqueous acetic acid (100 mL) was heated at 60 ꢁC for
1.5 h. The cooled mixture was evaporated under
vacuum and the last traces of acetic acid removed by co-
evaporation with toluene. Chromatography of the resi-
due on silica gel (7Â13 cm, 0–10% methanol/chloro-
form) gave the title compound (5.67 g, 78%) as a solid.
Recrystallization from dichloromethane gave a white
solid; mp 131–132 ꢁC. [a]D22 +22.7ꢁ (c 1.0, CHCl3).
1H NMR (400 MHz, CDCl3) d 1.35 (3H, t, J=7.5 Hz,
-SCH2CH3), 2.22 (1H, m, -OH), 2.50 (1H, d, J=4.4 Hz,
-OH), 2.73–2.87 (3H, m, OH and -SCH2CH3), 3.5–3.6
(2H, m, H-2 and H-5 overlapping), 3.61 (1H, m, H-3),
3.82 (3H, s, -OCH3), 3.84 (1H, m, H-6), 3.95 (1H, m, H-
6), 4.03 (1H, br s, H-4), 4.44 (1H, d, J=9.4 Hz, H-1),
4.63 and 4.92 (2H, 2d, J=10.8 Hz, -OCH2Ar), 6.91 (2H,
d, J=8.6 Hz, H-3 of 4-methoxybenzyl) and 7.34 (2H, d,
J=8.6 Hz, H-2 of 4-methoxybenzyl). IR (KBr) ꢀmax
(cmÀ1): 3500 and 3300 (broad, OH) and 1605 (aro-
matic). Anal. calcd for C16H24O6S: C, 55.80; H, 7.02; S,
9.31. Found: C, 55.64; H, 6.78; S, 9.23.
Ethyl 2-O-(3,3-di-tert-butyloxycarbonylpropyl)-3-O-ben-
zoyl-4,6-O-benzylidene-1-thio-ꢀ-D-galactopyranoside 20.
Ethyl 2-O-(3,3-di-tert-butyloxycarbonylpropyl)-4,6-O-
benzylidene-1-thio-b-d-galactopyranoside 19 (0.751 g,
1.35 mmol) in methylene chloride (8 mL) and pyridine
(7 mL) was reacted by the procedure used to synthesize
compound 18 and gave the title material (0.888 g, 99%)
as an oil. [a]2D2 +51ꢁ (c 1.1, CHCl3). 1H NMR
(400 MHz, CDCl3) d 1.36 (3H, t, J=7.6 Hz, -CH3), 1.37
and 1.42 (18H, 2s, 2Â-OtBu), 2.00 (2H, m, -CH2–
CH(CO2tBu)2), 2.82 (2H, m, -SCH2-), 3.37 (1H, dd,
J=7.9, 6.6 Hz, -CH(CO2tBu)2), 3.58 (1H, br s, H-5),
3.75 (1H, dt, J=9.2, 5.9 Hz, -OCH2-), 3.81–3.87 (1H, m,
-OCH2-), 3.87 (1H, dd, J=9.6, 9.6 Hz, H-2), 4.03 (1H,
dd, J=12.4, 1.6 Hz, H-6), 4.36 (1H, dd, J=12.4, 1.3 Hz,
H-6), 4.50 (1H, d, J=9.6 Hz, H-1), 4.53 (1H, d,
J=3.5 Hz, H-4), 5.12 (1H, dd, J=9.6, 3.5 Hz, H-3), 5.49
(1H, s, -OCHO-), 7.34–7.38, 7.43–7.48, 7.56–7.60 and
8.08–8.10 (10H, 4 sets of m, 2Â-C6H5). IR (film) ꢀmax
Ethyl 2-O-p-methoxybenzyl-4,6-di-O-benzylidene-1-thio-
ꢀ-D-galactopyranoside 23. Ethyl 2-O-p-methoxybenzyl-
1-thio-b-d-galactopyranoside 22 (6.11 g, 17.7 mmol) in

A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
1409
acetonitrile was reacted by the procedure used to syn-
thesize compound 17 and afforded the title compound
(4.72 g, 62%) as a colorless oil. [a]2D2 +14.1ꢁ (c 1.0,
CHCl3). 1H NMR (400 MHz, CDCl3) d 1.36 (3H, t,
J=7.4 Hz, -CH3), 2.52 (1H, d, J=8.3 Hz, -OH), 2.81
(2H, m, -SCH2-), 3.47 (1H, br d, H-5), 3.64 (1H, dd,
J=9.2, 9.2 Hz, H-2), 3.77 (1H, m overlapped by -OCH3,
H-3), 3.80 (3H, s, -OCH3), 4.03 (1H, dd, J=12.5,
1.7 Hz, H-6), 4.25 (1H, d, J=3.5 Hz, H-4), 4.35 (1H, dd,
J=12.5, 1.2 Hz, H-6), 4.44 (1H, d, J=9.5 Hz, H-1), 4.72
(1H, d, J=10.3 Hz, -OCH2Ar), 4.88 (1H, d, J=10.3 Hz,
-OCH2Ar), 5.57 (1H, s, -OCHO-), 6.87–6.92, 7.15–7.19,
7.25–7.32, 7.36–7.43 and 7.50–7.54 (9H, 5 sets of m,
aromatic H). IR (KBr) ꢀmax (cmÀ1) 3600–3250 (broad,
OH) and 1610, 1510 (aromatic C¼C). Anal. calcd for
C23H28O6S.0.2 H2O: C, 63.34; H, 6.56. Found: C, 63.42;
H, 6.40.
anoside 24 (1.5 g, 2.64 mmol) was reacted by the pro-
cedure used to synthesize compound 8 and afforded the
title compound (1.52 g, 82%) as a white solid. 1H NMR
(400 MHz, CDCl3) d 1.34 (3H, t, J=7.5 Hz, -CH3), 1.45
(18H, s, 2Â-OtBu), 1.58, 1.68 and 1.83 (3Â2H, -(CH2)3-),
2.80 (2H, m, -SCH2-), 3.10 (1H, t, J=7.5 Hz, -CH(CO2-
tBu)2), 3.42 (1H, br s, H-5), 3.45 (1H, dd, J=9.2,
3.5 Hz, H-3), 3.59 (1H, dt, J=9.2, 6.6 Hz, -OCH2-), 3.71
(1H, dt, J=9.2, 6.7 Hz, -OCH2-), 3.76–3.84 (1H, m
overlapped by -OCH3, H-2), 3.81 (3H, s, -OCH3), 4.03
(1H, dd, J=12.3, 1.5 Hz, H-6), 4.30 (1H, d, J=3.4 Hz,
H-4), 4.34 (1H, dd, J=12.3, 1.2 Hz, H-6), 4.43 (1H, d,
J=9.6 Hz, H-1), 4.74 (1H, d, J=9.9 Hz, -CH2OAr), 4.78
(1H, d, J=9.9 Hz, -OCH2Ar), 5.54 (1H, s, -OCHO-),
6.85–6.92, 7.30–7.40 and 7.49–7.54 (9H, 3 sets of m,
aromatic H). IR (CH2Cl2) ꢀmax (cmÀ1) 3050, 2980 (C–
H), 1740, 1720 (C¼O). Anal. calcd for C38H54O10S: C,
64.93; H, 7.74. Found: C, 64.94; H, 7.64.
Ethyl 2-O-p-methoxybenzyl-4,6-di-O-benzylidene-3-O-
(4-bromobutyl)-1-thio-ꢀ-D-galactopyranoside 24. Ethyl
2-O-p-methoxybenzyl-4,6-di-O-benzylidene-1-thio-b-d-
galactopyranoside 23 (3.5 g, 8.1 mmol) was reacted by
the procedure used to synthesize compound 7 and affor-
ded the title compound (2.7 g, 59%) as an oil. 1H NMR
(400 MHz, CDCl3) d 1.35 (3H, t, J=7.5 Hz, -CH3), 1.76–
1.82 and 1.96–2.03 (2Â2H, 2 sets of m, -(CH2)2-), 2.81
(2H, m, -SCH2-), 3.38–3.44 (3H, m, H-5 and -CH2Br),
3.46 (1H, dd, J=9.1, 3.2 Hz, H-3), 3.61 (1H, dt, J=9.2,
6.0 Hz, -OCH2-), 3.75 (1H, dt, J=9.2, 6.1 Hz, -OCH2-),
3.78 (1H, br dd, H-2), 3.81 (3H, s, -OCH3), 4.03 (1H,
dd, J=12.3, 1.6 Hz, H-6), 4.32 (1H, d, J=3.2 Hz, H-4),
4.36 (1H, dd, J=12.3, 1.3 Hz, H-6), 4.44 (1H, d,
J=9.6 Hz, H-1), 4.72 (1H, d, J=9.8 Hz, -OCH2Ar),
4.81 (1H, d, J=9.8 Hz, -OCH2Ar), 5.55 (1H, s, -OCHO-),
6.87–6.90, 7.33–7.41 and 7.52–7.55 (9H, 3 sets of m,
aromatic H). IR (CH2Cl2) ꢀmax (cmÀ1): 3050, 2980 (C–
H), 1610, 1515 (aromatic C¼C). Anal. calcd for
C27H35O6SBr: C, 57.14; H, 6.22. Found: C, 57.38; H,
6.14.
Ethyl 2-O-p-methoxybenzyl-4,6-di-O-benzylidene-3-O-[4-
(2,2-di-tert-butyloxycarbonylethyl)benzyl]-1-thio-ꢀ-D-gal-
actopyranoside 27. Ethyl 2-O-p-methoxybenzyl-4,6-di-
O-benzylidene-3-O-(4-bromobutyl)-1-thio-b-d-galacto-
pyranoside 24 (0.714 g, 3.3 mmol) was reacted by the
procedure used to synthesize compound 8 and afforded
the title compound (0.922 g, 82%) as a white solid.
1H NMR (400 MHz, CDCl3) d 1.34 (3H, t, J=7.5 Hz,
-SCH2CH3), 1.42 (2Â9H, 2 s, 2Â-OtBu), 2.80 (2H, m,
-SCH2-), 3.13 (2H, d, J=7.8 Hz, -CH2-CH(CO2tBu)2),
3.34 (1H, s, H-5), 3.45 (1H, t, J=7.8 Hz, -CH(CO2-
tBu)2), 3.55 (1H, dd, J=9.2, 3.4 Hz, H-3), 3.82 (3H, s,
-OCH3), 3.87 (1H, dd, J=9.3, 9.3 Hz, H-2), 3.97 (1H, d,
J=12.3 Hz, H-6), 4.12 (1H, d, J=3.3 Hz, H-4), 4.31
(1H, d, J=12.3 Hz, H-6), 4.42 (1H, d, J=9.6 Hz, H-1),
4.67–4.84 (4H, m, 2Â-OCH2Ar), 6.87–6.89, 7.16–7.18,
7.27–7.41 and 7.54–7.56 (13H, 4 sets of m, aromatic H).
IR (CH2Cl2) ꢀmax (cmÀ1) 3050, 2980 (C–H), 1740, 1720
(C¼O).
Synthesis of (2S,3R,4E)-3-benzoyloxy-2-hexadecanoyla-
mino-1-[2-O-(5,5-di-carboxypentyl)-ꢁ-D-galactopyranosy-
loxy]-4-octadecene 33a
Ethyl 2-O-p-methoxybenzyl-4,6-di-O-benzylidene-3-O-
(4-bromomethylphenyl)-1-thio-ꢀ-D-galactopyranoside 25.
Ethyl 2-O-p-methoxybenzyl-4,6-di-O-benzylidene-1-thio-
b-d-galactopyranoside 23 (1.03 g, 2.38 mmol)) was reac-
ted by the procedure used to synthesize compound 7
except that dibromoxylene was used instead of dibro-
mobutane. This afforded the title compound (0.92 g,
61%) as an oil. 1H NMR (400 MHz, CDCl3) d 1.35 (3H,
t, J=7.5 Hz, -SCH2CH3), 2.81 (2H, m, -SCH2-), 3.37
(1H, s, H-5), 3.57 (1H, dd, J=9.2, 3.5 Hz, H-3), 3.82
(3H, s, -OCH3), 3.88 (1H, dd, J=9.4, 9.4 Hz, H-2), 3.98
(1H, dd, J=12.3, 1.6 Hz, H-6), 4.18 (1H, d, J=3.4 Hz,
H-4), 4.32 (1H, dd, J=12.3, 1.3 Hz, H-6), 4.43 (1H, d,
J=9.6 Hz, H-1), 4.50 (2H, s, -CH2Br), 4.75 and 4.84
(2Â 1H, 2 d, J=9.8 Hz, -OCH2Ar), 4.76 (2H, s, -OCH2-
of p-bromomethylbenzyl), 5.49 (1H, s, -OCHO-), 6.86–
6.89, 7.34–7.42 and 7.54–7.56 (13H, 3 sets of m, aro-
matic H). IR (CH2Cl2) ꢀmax (cmÀ1) 3050, 2980 (C-H).
(2S,3R,4E)-3-Benzoyloxy-2-azido-1-[2-O-(5,5-di-tert-
butyloxycarbonylpentyl)-3,4-O-isopropylidene-6-O-tert-
butyldimethylsilyl-ꢁ-D-galactopyranosyloxy]-4-octade-
cene 29a and (2S,3R,4E)-3-benzoyloxy-2-azido-1-[2-O-
(5,5-di-tert-butyloxycarbonylpentyl)-3,4-O-iso-
propylidene-6-O-tert-butyldimethylsilyl-b-D-galactopyr-
anosyloxy]-4-octadecene 29b. A solution of ethyl 3,4-O-
isopropylidene-6-O-tert-butyldimethylsilyl-2-O-(5,5-di-
tert-butyloxycarbonylpentyl)-1-thio-b-d-galactopyrano-
side 8 (1.0 g, 1.54 mmol), 2,6-di-tert-butyl-4-methylpyr-
idine (0.627 g, 3.08 mmol) and (2S,3R,4E)-2-azido-3-
benzoyloxy-4-octadecen-1-ol 2827 (0.640 g, 1.48 mmol)
in toluene (12 mL) was stirred with molecular sieves (4
A, previously heated with a Bunsen flame) for 1 h at
22 ꢁC. Then dimethyl(methylthio)sulfonium triflate
(0.765 g, 3.08 mmol) was added in and stirring was con-
tinued for 1 h. The mixture was diluted with ethyl ace-
tate (12 mL) and filtered through Celite. The filtrate was
washed with 1 M aqueous sodium bicarbonate (12 mL)
and brine (12 mL), dried over anhydrous magnesium
sulfate, filtered and concentrated. The residue was
Ethyl 2-O-p-methoxybenzyl-4,6-di-O-benzylidene-3-O-
(5,5-di-tert-butyloxycarbonylpentyl)-1-thio-ꢀ-D-galacto-
pyranoside 26. Ethyl 2-O-p-methoxybenzyl-4,6-di-O-
benzylidene-3-O-(4-bromobutyl)-1-thio-b-d-galactopyr-

1410
A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
purified by silica gel column chromatography (5–30%
ethyl acetate/hexane) and gave the a-anomer (0.595 g,
39%) and the b-anomer (0.321 g, 21%) of the title
compound as oils. a-Anomer: 1H NMR (400 MHz,
CDCl3) d 0.08 (6H, s, -Si(CH3)2), 0.89 (3H, t, J=7.0 Hz,
-CH3), 0.91 (9H, s, -SitBu), 1.25–1.51 and 1.58–1.66
(26H, 2 sets of m, -(CH2)11- and -(CH2)2-), 1.45 (18H, s,
2Â-OtBu), 1.33 and 1.51 (2Â3H, 2 s, -C(CH3)2-), 1.81
and 2.07 (2Â2H, 2 ap q,=CH-CH2- and -CH2–
CH(CO2tBu)2), 3.11 (1H, t, J=7.5 Hz, -CH(CO2tBu)2),
3.44 (1H, dd, J=6.9, 3.4 Hz, H-20), 3.52 (1H, dd,
J=10.8, 8.0 Hz, H-1), 3.61 (2H, t, J=6.7 Hz, -OCH2-),
3.78 (1H, dd, J=10.0, 6.6 Hz, H-60), 3.83–3.88 (2H, m,
H-1 and H-60), 3.99 (1H, m, H-2), 4.04 (1H, td, J=6.5,
2.0 Hz, H-50), 4.21 (1H, dd, J=5.6, 2.2 Hz, H-40), 4.20–
4.26 (1H, m overlapped by H-40, H-30), 4.91 (1H, d,
J=3.4 Hz, H-10), 5.54–5.61 (2H, m, H-3 and H-4), 5.93
(1H, dt, J=14.1 and 6.8 Hz, H-5), 7.44–7.48, 7.56–7.60
and 8.06–8.08 (5H, 3 sets of m, aromatic H). IR
(CH2Cl2) ꢀmax (cmÀ1): 3050, 2980, 2930, 2855 (C-H),
2105 (-N3), 1720 (C¼O). Anal. calcd for
C55H93N3O13Si: C, 64.99; H, 9.22; N, 4.13. Found: C,
65.05; H, 8.97; N, 4.23. b-Anomer: 1H NMR (400 MHz,
CDCl3) d 0.07 (6H, s, -Si(CH3)2), 0.89 (12H, m, -SitBu
and -CH3), 1.25–1.52 and 1.61–1.68 (26H, 2 sets of m,
-(CH2)11- and -(CH2)2-), 1.33 and 1.53 (2Â3H, 2 s,
-C(CH3)2-), 1.45 (18H, s, 2Â-OtBu), 1.83 (2H, ap q,
J=7.8 Hz, -CH2-CH(CO2tBu)2), 2.07 (2H, ap q,
J=6.9 Hz,=CH-CH2-), 3.13 (1H, br t, H-20), 3.25 (1H, t,
J=7.4 Hz, -CH(CO2tBu)2), 3.57 (1H, dd, J=10.1,
5.2 Hz, H-1), 3.69 (1H, dt, J=9.5, 6.8 Hz, -OCH2-),
3.66–3.82 and 3.98–4.01 (3H, 2 sets of m, H-50, H-2 or
-OCH2-), 3.81 (1H, dd, J=9.8, 6.1 Hz, H-60), 3.86 (1H,
dd, J=9.8, 7.1 Hz, H-60), 3.93 (1H, dd, J=10.1, 7.6 Hz,
H-1), 4.04 (1H, br t, H-30), 4.16 (1H, dd, J=5.5, 1.8 Hz,
H-40), 4.20 (1H, d, J=7.9 Hz, H-10), 5.57 (1H, dd,
J=15.1, 8.0 Hz, H-4), 5.63 (1H, dd, J=8.0, 4.0 Hz, H-3),
5.94 (1H, dt, J=15.1, 6.9 Hz, H-5), 7.44–7.47, 7.56–7.59
and 8.06–8.08 (5H, 3 sets of m, aromatic H). IR
(CH2Cl2) ꢀmax (cmÀ1) 3050, 2980, 2930, 2860 (C-H),
2105 (-N3), 1720 (C¼O). Anal. calcd for
C55H93N3O13Si: C, 64.99; H, 9.22; N, 4.13. Found: C,
65.30; H, 9.08; N, 4.16.
then diluted with ethyl acetate (ꢀ15 mL). The organic
layer was separated, washed with 1 M aqueous sodium
bicarbonate (ꢀ15 mL) and brine (ꢀ15 mL), dried over
anhydrous magnesium sulfate, filtered and con-
centrated. The residue was purified by silica gel column
chromatography (20–50% ethyl acetate/hexane) to
afford the title compound (0.406 g, 73%) as a beige solid.
1H NMR (400 MHz, CDCl3) d 0.07 (6H, s, -Si(CH3)2),
0.87–0.90 (15H, m, -SitBu and 2Â -CH3), 1.24–1.49
and 1.60–1.63 (76H, m, -(CH2)13-, -(CH2)11-, -(CH2)2-,
2Â-OtBu and -C(CH3)2-), 1.83 and 2.02 (2Â2H, 2 qa,
-CH2-CH(CO2tBu)2 and ¼CH–CH2-), 2.20 (2H, m,
-NHCOCH2-), 3.12 (1H, t, J=7.5 Hz, -CH(CO2tBu)2),
3.40 (1H, dd, J=6.3, 3.4 Hz, H-20), 3.60 (2H, t,
J=6.6 Hz, -OCH2-), 3.71–3.77, 3.80–3.86 and 4.18–4.21
(3Â2H, 3 sets of m, H-1, H-60, H-40 and H-50), 4.02 (1H,
dd, J=6.4, 6.4 Hz, H-30), 4.49 (1H, m, H-2), 4.80 (1H, d,
J=3.3 Hz, H-10), 5.53 (1H, dd, J=15.1, 7.5 Hz, H-4),
5.60 (1H, t, J=7.3 Hz, H-3), 5.87 (1H, dt, J=15.1,
6.9 Hz, H-5), 6.11 (1H, d, J=9.1 Hz, -NH-), 7.43–7.47,
7.55–7.59 and 8.03–8.05 (5H, 3 sets of m, aromatic H).
IR (CH2Cl2) ꢀmax (cmÀ1) 3440 (N–H), 3050, 2980, 2930,
2860 (C–H), 1720, 1675 (C¼O). Anal. calcd for
C71H125NO13Si: C, 69.40; H, 10.25; N, 1.14. Found: C,
69.40; H, 10.13; N, 1.27.
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-
O-(5,5-di-carboxypentyl)-a-D-galactopyranosyloxy]-4-
octadecene 33a. (2S,3R,4E)-3-Benzoyloxy-2-hexadecan-
oylamino-1-[2-O-(5,5-di-tert-butyloxycarbonylpentyl)-3,4-
O-isopropylidene-6-O-tert-butyldimethylsilyl-a-d-galac-
topyranosyloxy]-4-octadecene 30a (0.376 g, 0.30 mmol)
was treated with 90% aqueous trifluoroacetic acid
(4 mL) at 5 ꢁC. This mixture was stirred for 5 min at
5 ꢁC and then the ice-bath was removed and this was
stirred for 10 min. Toluene was added and the mixture
was evaporated under vaccuo and coevaporated with
toluene (2Â). This was repeated until there was no
starting material left. The residue was dried in vaccuo,
dissolved in chloroform and filtered on Millex LCR
carthridge. The solution was triturated with acetonitrile.
The residue was recrystallized form chloroform/aceto-
nitrile to give the title compound (0.175 g, 61%) as a
white solid. 1H NMR (400 MHz, pyridine-d5) d 0.85
(6H, t, J=6.7 Hz, 2Â-CH3), 1.21–1.40, 1.72–1.78 and
1.84–1.89 (52H, 3 sets of m, -(CH2)13-, -(CH2)11- and
-(CH2)2-), 2.02 and 2.33 (2Â2H, 2 ap q, -CH2-
CH(CO2H)2 and ¼CH–CH2-), 2.51 (2H, t,
J=7.4 Hz, -NHCOCH2-), 3.66 (1H, m, -OCH2-),
3.78 (1H, m, -OCH2-), 3.83 (1H, t, J=7.3 Hz,
-CH(CO2H)2), 4.18–4.22, 4.37–4.42 and 4.43–4.50 (2H
and 6H, H-1, H-60, H-50, H-40, H-30 and H-20), 5.20 (1H,
m, H-2), 5.44 (1H, d, J=3.5 Hz, H-10), 5.92 (1H, dd,
J=15.4, 7.4 Hz, H-4), 6.09 (1H, dt, J=15.4, 6.7 Hz, H-
5), 6.25 (1H, br t, H-3), 7.41–7.51 and 8.25–8.27 (5H, 2
sets of m, aromatic H), 8.79 (1H, d, J=8.9 Hz, -NH-).
IR (nujol) ꢀmax (cmÀ1) 3700–3100 (N–H, O–H), 2930,
2860 (C–H), 1720, 1645 (C¼O). MS (ESI-): 960.6
(MÀH)-. MS (ESI+): 962.7 (MH+), 984.6 (M+Na)+.
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-
O-(5,5-di-tert-butyloxycarbonylpentyl)-3,4-O-iso-
propylidene-6-O-tert-butyldimethylsilyl-ꢁ-D-galactopyr-
anosyloxy]-4-octadecene 30a. A solution of (2S,3R,4E)-
3-benzoyloxy-2-azido-1-[2-O-(5,5-di-tert-butyloxycar-
bonylpentyl)-3,4-O-isopropylidene-6-O-tert-butyldi-
methylsilyl-a-d-galactopyranosyloxy]-4-octadecene 29a
(0.465 g, 0.45 mmol) in pyridine (15 mL) and water
(3 mL) was treated with a stream of hydrogen sulfide for
ꢀ10 min. The solution was stirred for 12 h, then hydro-
gen sulfide was bubbled again for ꢀ10 min. The reac-
tion mixture was stirred for another 12 h. The solvents
were evaporated under vacuum and the residue was co-
evaporated with toluene (2Â). The residue was then
dissolved in tetrahydrofuran (15 mL) and an aqueous
solution of sodium acetate (50%, 3 mL) was added fol-
lowed by palmitoyl chloride (184 mL, 0.603 mmol) at
0 ꢁC. The mixture was stirred for 30 min at 22 ꢁC and
Preparation of the sodium salt of the title compound:
The diacid from the above procedure (0.159 g,
0.170 mmol) was dissolved in dioxane (10 mL) and fil-

A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
1411
1
tered on Millex LCR 0.5 m. This solution was treated
with an aqueous solution of sodium bicarbonate (30 mg,
0.348 mmol in 5 mL of water). The resulting suspension
was then diluted with water to get a slightly cloudy
solution which was lyophilized to give the sodium salt of
the title compound (0.172 g, 100%) as a white fluffy
solid. 1H NMR (400 MHz, CD3OD) d 0.83 (6H, t,
J=6.8 Hz, 2Â-CH3), 1.19–1.38, 1.55, 1.77 (48H, 4H and
2H, 3 sets of m, -(CH2)13-, -(CH2)11-, -(CH2)2- and
-CH2-CH(CO2Na)2), 2.00 (2H, dt, J=6.9, 6.9 Hz,
¼CH-CH2-), 2.15 (2H, t, -NHCOCH2-), 3.03 (1H, t,
J=7.2 Hz, -CH(CO2Na)2), 3.44 (1H, m, -OCH2-), 3.50
(1H, dd, J=9.9, 3.6 Hz, H-20), 3.56–3.65 and 3.73–3.76
(4H and 2H, 2 sets of m, H-1, H-60, -OCH2- and H-30),
3.69 (1H, t, J=6.0 Hz, H-50), 3.80 (1H, d, J=3.1 Hz,
H-40), 4.36 (1H, m, H-2), 4.85 (1H, d, J=3.5 Hz, H-10),
5.45 (1H, dd, J=15.0 and 7.9 Hz, H-4), 5.52 (1H, br t,
H-3), 5.84 (1H, dt, J=15.0 and 6.9 Hz, H-5), 7.40–7.44,
7.52–7.56 and 7.95–7.97 (5H, 3 sets of m, aromatic H).
IR (nujol) ꢀmax (cmÀ1): 3700–3100 (N–H, O–H), 2920,
2850 (C–H), 1710, 1645, 1585 (C¼O).
78%) as a white solid. Diacid: H NMR (400 MHz,
pyridine-d5) d 0.85 (6H, t, J=6.8 Hz, 2Â-CH3), 1.23–1.44
and 1.79–1.91 (52H, 2 sets of m, -(CH2)13-, -(CH2)11- and
-(CH2)2-), 2.04 (2H, dt, J=6.9, 6.9 Hz,=CH-CH2-),
2.40 (2H, ap q, -CH2-CH(CO2H)2), 2.48 (2H,
t, J=7.4 Hz, -NHCOCH2-), 3.90 (1H, t, J=7.4 Hz,
-CH(CO2H)2), 3.92–4.07 (4H, m, H-20, H-30, H-50 and
-OCH2-), 4.17–4.24 (1H, m, -OCH2-), 4.23 (1H, dd,
J=10.6, 5.1 Hz, H-1), 4.34 (1H, dd, J=10.9, 6.0 Hz, H-
60), 4.40 (1H, dd, J=10.9, 6.0 Hz, H-60), 4.48 (1H, dd,
J=10.6, 6.3 Hz, H-1), 4.49 (1H, br s, H-40), 4.79 (1H, d,
J=7.0 Hz, H-10), 5.20 (1H, m, H-2), 5.93 (1H, dd,
J=15.4, 7.4 Hz, H-4), 6.11 (1H, dt, J=15.4, 6.7 Hz, H-
5), 6.26 (1H, br t, H-3), 7.36–7.40, 7.45–7.55 and 8.23–
8.25 (5H, 3 sets of m, aromatic H), 8.52 (1H, d,
J=8.7 Hz, -NH-). IR (nujol) ꢀmax (cmÀ1) 3700–2400
(O–H, N–H), 2920, 2850 (C–H), 1710, 1640 (C¼O). MS
(ESIÀ): 960.6 (MÀH)À. MS (ESI+): 962.6 (MH+),
984.6 (M+Na)+. Sodium salt: 1H NMR (400 MHz,
CD3OD) d 0.89 (6H, t, J=6.8 Hz, 2Â-CH3), 1.25–1.53,
1.59–1.64 and 1.7–1.9 (54H, 3 sets of m, -(CH2)13-,
-(CH2)11-, -CH2-CH(CO2Na)2 and -(CH2)2-), 2.08 (2H,
dt, J=6.9, 6.9 Hz, ¼CH–CH2-), 2.20 (2H, m,
-NHCOCH2-), 3.09 (1H, t, J=6.8 Hz, -CH(CO2Na)2),
3.31–3.29 (1H, m, H-20), 3.43 (1H, dd, J=6.0, 6.0 Hz,
H-50), 3.48 (1H, dd, J=9.5, 3.3 Hz, H-30), 3.64–3.66
(3H, m, -OCH2- and H-60), 3.76 (1H, dd, J=10.6,
5.3 Hz, H-1), 3.80 (1H, d, J=3.2 Hz, H-40), 3.79–3.86
(1H, m, -OCH2-), 3.96 (1H, dd, J=10.6, 6.3 Hz, H-1),
4.25 (1H, d, J=7.7 Hz, H-10), 4.47 (1H, m, H-2), 5.55
(1H, dd, J=15.2, 7.7 Hz, H-4), 5.63 (1H, dd, J=7.5,
6.0 Hz, H-3), 5.89 (1H, dt, J=15.2, 6.9 Hz, H-5), 7.44–
7.48, 7.57–7.61 and 8.00–8.02 (5H, 3 sets of m, aromatic
H). IR (nujol) ꢀmax (cmÀ1) 3700-2500 (O–H, N–H),
2920, 2850 (C–H), 1710, 1640, 1575 (C¼O).
Synthesis of (2S,3R,4E)-3-benzoyloxy-2-hexadecanoyla-
mino-1-[2-O-(5,5-di-carboxypentyl)-ꢀ-D-galactopyranosy-
loxy]-4-octadecene 33b
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-
O-(5,5-di-tert-butyloxycarbonylpentyl)-3,4-O-iso-
propylidene-6-O-tert-butyldimethylsilyl-ꢀ-D-galactopyr-
anosyloxy]-4-octadecene 30b. (2S,3R,4E)-3-Benzoyloxy-
2-azido-1-[2-O-(5,5-di-tert-butyloxycarbonylpentyl)-3,4-
O-isopropylidene-6-O-tert-butyldimethylsilyl-b-d-galac-
topyranosyloxy]-4-octadecene 29b (0.473 g, 0.465 mmol)
was reacted by the procedure used to synthesize com-
pound 30a and afforded the title compound (0.396 g,
69%) as a slighlty yellow oil. 1H NMR (400 MHz,
CDCl3) d 0.04 and 0.05 (6H, 2 s, -Si(CH3)2), 0.88–0.91
(15H, m, -SitBu and 2Â-CH3), 1.32 and 1.51 (2Â3H, 2 s,
-C(CH3)2-), 1.24–1.39 and 1.47–1.67 (52H, 2 sets of m,
-(CH2)2-, -(CH2)13- and -(CH2)11-), 1.47 (18H, s, 2Â
-OtBu), 1.80 (2H, dt, J=7.8, 7.8 Hz, -CH2-
CH(CO2tBu)2), 2.02 (2H, dt, J=7.0, 7.0 Hz, ¼CH-CH2-),
2.15 (2H, m, -NHCOCH2-), 3.11 (1H, t, J=7.5 Hz,
-CH(CO2tBu)2), 3.24 (1H, br dd, H-20), 3.58–3.77 and
4.15–4.18 (5H and 1H, 2 sets of m, -OCH2-, H-6, H-50 and
H-1), 3.81 (1H, dd, J=9.5, 7.3 Hz, H-1 or H-60), 4.04 (1H,
dd, J=5.7, 6.5 Hz, H-30), 4.16 (1H, dd, J=5.7, 1.4 Hz, H-
40), 4.17 (1H, d, J=7.8 Hz, H-10), 4.46 (1H, m, H-2), 5.51
(1H, dd, J=15.1, 7.4 Hz, H-4), 5.58 (1H, br t, H-3), 5.86
(1H, dt, J=15.1, 6.9 Hz, H-5), 6.29 (1H, d, J=9.1 Hz,
-NH-), 7.42–7.45, 7.54–7.57 and 8.03–8.06 (5H, 3 sets of
m, aromatic H). IR (CH2Cl2) ꢀmax (cmÀ1) 3690 (N-H),
3050, 2980, 2930, 2860 (C–H), 1720, 1670 (C¼O). Anal.
calcd for C71H125NO13Si: C, 69.40; H, 10.25; N, 1.14.
Found: C, 69.70; H, 10.31; N, 1.24.
Synthesis of (2S,3R,4E)-3-benzoyloxy-2-hexadecanoyl-
amino-1-[2-O-(3,3-di-carboxypropyl)-ꢁ-D-galactopyrano-
syloxy]-4-octadecene 34a. (2S,3R,4E)-3-Benzoyloxy-2-
azido-1-[2-O-(3,3-di-tert-butyloxycarbonylpropyl)-3,4-O-
isopropylidene-6-O-tert-butyldimethylsilyl-ꢁ-D-galacto-
pyranosyloxy]-4-octadecene 31a and (2S,3R,4E)-3-ben-
zoyloxy-2-azido-1-[2-O-(3,3-di-tert-butyloxycarbonylpro-
pyl)-3,4-O-isopropylidene-6-O-tert-butyldimethylsilyl-ꢀ-
D-galactopyranosyloxy]-4-octadecene 31b. Ethyl 3,4-O-
isopropylidene-6-O-tert-butyldimethylsilyl-2-O-(3,3-di-
tert-butyloxycarbonylpropyl)-1-thio-b-d-galactopyrano-
side 12 (0.81 g, 1.30 mmol) and (2S,3R,4E)-2-azido-3-
benzoyloxy-4-octadecen-1-ol 28 (0.375 g, 0.87 mmol)
were reacted by the procedure used to synthesize com-
pounds 29a and b except that dioxane was used as sol-
vent. This afforded the a-anomer (0.155 g, 18%) and the
b-anomer (0.050 g, 6%) of the title compound as oils.
1
ꢁ-Anomer: H NMR (400 MHz, CDCl3) d 0.08 (6H, m,
-Si(CH3)2), 0.89 (3H, t overlapped by -SitBu, -CH3),
0.91 (9H, s, -SitBu), 0.92–1.40 (22H, m, -(CH2)11-), 1.33
(3H, s, -C(CH3)2), 1.45 (18H, s, 2Â-OtBu), 1.50 (3H, s,
-C(CH3)2), 2.05–2.09 (4H, m, ¼CH-CH2- and -CH2-
CH(CO2tBu)2), 3.43–3.47 (2H, m, -CH(CO2tBu)2 and
H-2), 3.53 (1H, dd, J=10.7, 8.0 Hz, H-1), 3.63 (1H, dt,
J=9.8, 6.0 Hz, -OCH2-), 3.72 (1H, dt, J=9.8, 6.0 Hz,
-OCH2-), 3.78 (1H, dd, J=10.0, 6.6 Hz, H-60), 3.84–
3.88, 4.00–4.05 and 4.21–4.27 (3Â1H, 3 sets of m, H-1,
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-
O-(5,5-di-carboxypentyl)-b-D-galactopyranosyloxy]-4-octa-
decene 33b. (2S,3R,4E)-3-Benzoyloxy-2-hexadecanoyl-
amino-1-[2-O-(5,5-di-tert-butyloxycarbonylpentyl)-3,4-O-
isopropylidene-6-O-tert-butyldimethylsilyl-b-d-galacto-
pyranosyloxy]-4-octadecene 30b (0.358 g, 0.29 mmol)
was reacted by the procedure used to synthesize com-
pound 33a and afforded the title compound (0.217 g,

1412
A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
H-50 and H-60), 3.85 (1H, dd, J=6.5, 3.3 Hz, H-20), 4.03
(1H, dd, J=6.5, 2.2 Hz, H-30), 4.22 (1H, dd, J=5.6,
2.2 Hz, H-40), 4.91 (1H, d, J=3.4 Hz, H-10), 5.55–5.60
(2H, m, H-3 and H-4), 5.93 (1H, br dt, H-5), 7.44–7.48,
7.56–7.60 and 8.06–8.08 (5H, 3 sets of m, aromatic H).
2.00 (2Â2H, 2 sets of m, -CH2-CH(CO2H)2 and -CH2-),
2.53 (2H, t, J=7.4 Hz, -NHCOCH2-), 2.76 (2H, dt,
J=6.6, 6.6 Hz,=CH–CH2-), 4.07 (1H, dt, J=9.3,
6.2 Hz, -OCH2-), 4.16 (1H, dt, J=9.3, 6.0 Hz, -OCH2-),
4.23 (1H, dd, J=10.7, 6.6 Hz, H-60), 4.28–4.35 and
4.43–4.55 (7H, 2 sets of m, H-1, H-20, H-30, H-40, H-60,
H-50 and -CH(CO2H)2), 4.38 (1H, dd, J=10.4, 4.8 Hz,
H-1), 5.17–5.22 (1H, m, H-2), 3.52 (1H, d, J=3.5 Hz,
H-10), 5.92 (1H, dd, J=15.4, 7.3 Hz, H-4), 6.08 (1H, dt,
J=15.4, 6.6 Hz, H-5), 6.29 (1H, br t, H-3), 7.36–7.40,
7.45–7.49 and 8.24–8.28 (5H, 3 sets of m, aromatic H),
1
ꢀ-Anomer: H NMR (400 MHz, CDCl3) d 0.06 (6H, s,
-Si(CH3)2), 0.89 (12H, m, -SitBu and -CH3), 1.24 (20H,
m, -(CH2)10-), 1.31 (3H, s, -C(CH3)2), 1.34–1.50 (2H, m,
-CH2-), 1.44 and 1.45 (18H, 2 s , 2Â-OtBu), 1.51 (3H, s,
-C(CH3)2), 2.04–2.14 (4H, m, ¼CH-CH2- and -CH2-
CH(CO2tBu)2), 3.24 (1H, t, J=7.3 Hz, -CH(CO2tBu)2),
3.42 (1H, br dd, H-20), 3.61 (1H, dd, J=10.3, 5.0 Hz, H-
1), 3.70–3.76 (2H, m, H-2 and -OCH2-), 3.78–3.93 (1H,
m, -OCH2-), 3.80 (1H, dd, J=10.0, 5.8 Hz, H-60), 3.86
(1H, dd, J=10.0, 7.2 Hz, H-60), 3.90 (1H, dd, J=10.2,
7.7 Hz, H-1), 4.01–4.06 (2H, m, H-30 and H-50), 4.16
(1H, dd, J=5.6, 1.8 Hz, H-40), 4.20 (1H, d, J=7.8 Hz,
H-10), 5.56 (1H, dd, J=14.8, 8.0 Hz, H-4), 5.62 (1H, dd,
J=8.0, 4.0 Hz, H-3), 5.93 (1H, dt, J=14.8, 6.7 Hz, H-5),
7.42–7.46, 7.55–7.58 and 8.05–8.07 (5H, 3 sets of m,
aromatic H).
1
8.80 (1H, d, J=8.6 Hz, -NH-). Sodium salt: H NMR
(400 MHz, CD3OD) d 0.83 (6H, m, 2Â-CH3), 1.21–1.39
(46H, m, -(CH2)13- and -(CH2)10), 1.53 (2H, m, -CH2-),
1.98–2.19 (6H, m, -NHCOCH2-, ¼CH-CH2- and -CH2-
CH(CO2Na)2), 5.21 (1H, t, J=6.9 Hz, -CH(CO2Na)2),
3.46–3.80 (10H, m, H-1, H-20, H-30, H-40, H-50, H-60 and
-OCH2-), 4.35 (1H, m, H-2), 4.88 (1H, d, J=2.5 Hz, H-
10), 5.46 (1H, dd, J=14.8, 8.0 Hz, H-4), 5.53 (1H, br t,
H-3), 5.84 (1H, dt, J=14.8, 6.4 Hz, H-5), 7.39–7.43,
7.51–7.55 and 7.96–7.98 (5H, 3 sets of m, aromatic H).
IR (film) ꢀmax (cmÀ1) 3700–3000 (NH), 2920, 2850 (C–
H), 1720, 1650, 1600 (C¼O).
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-O-
(3,3-di-tert-butyloxycarbonylpropyl)-3,4-O-isopropylidene-
6-O-tert-butyldimethylsilyl-ꢁ-D-galactopyranosyloxy]-4-
octadecene 32a. (2S,3R,4E)-3-Benzoyloxy-2-azido-1-[2-
O-(3,3-di-tert-butyloxycarbonylpropyl)-3,4-O-isopropyl-
idene-6-O-tert-butyldimethylsilyl-a-d-galactopyranosy-
loxy]-4-octadecene 31a (0.230 g, 0.233 mmol) was reac-
ted by the procedure used to synthesize compound 30a
and afforded the title compound (0.168 g, 60%) as an
oil. [a]2D2 +40ꢁ (c 1.0, CHCl3). 1H NMR (400 MHz,
CDCl3) d 0.07 (6H, s, -Si(CH3)2), 0.90 (15H, m, -SitBu
and 2Â-CH3), 1.25–1.27 (46H, m, -(CH2)10 and
-(CH2)13-), 1.31 (3H, s, -C(CH3)2), 1.43 and 1.45 (18H, 2
s, -OtBu), 1.46 (3H, s, -C(CH3)2), 1.49–1.62 (4H, m,
-CH2- and -CH2-CH(CO2tBu)2), 2.00–2.09 and 2.13–
2.22 (2Â2H, 2 sets of m, -NHCOCH2- and ¼CH–CH2-),
3.40 (1H, dd, J=6.3, 3.3 Hz, H-20), 3.46 (1H, t,
J=7.5 Hz, -CH(CO2tBu)2), 3.57–3.70 (3H, m, -OCH2-
and H-1), 3.74 (1H, dd, J=9.8, 6.3 Hz, H-60), 3.83 (1H,
dd, J=9.8, 7.1 Hz, H-60), 3.88 (1H, dd, J=10.6, 3.6 Hz,
H-1), 4.00 (1H, br dd, H-50), 4.18–4.21 (2H, m, H-30 and
H-40), 4.49 (1H, m, H-2), 4.79 (1H, d, J=3.2 Hz, H-10),
5.54 (1H, dd, J=15.3, 7.5 Hz, H-4), 5.65 (1H, br t, H-3),
5.89 (1H, dt, J=15.3, 6.7 Hz, H-5), 6.59 (1H, d,
J=8.8 Hz, -NH-), 7.42–7.46, 7.53–7.58 and 8.03–8.05
Synthesis of (2S,3R,4E)-3-benzoyloxy-2-hexadecanoyl-
amino-1-[2-O-(5,5-dicarboxypentyl)-3-O-benzoyl-ꢁ-D-ga-
lactopyranosyloxy]-4-octadecene 41a. (2S,3R,4E)-3-
Benzoyloxy-2-azido-1-[2-O-(5,5-di-tert-butyloxycarbonyl-
pentyl)-3-O-benzoyl-4,6-O-benzylidene-ꢁ-D-galactopyra-
nosyloxy]-4-octadecene 36a and (2S,3R,4E)-3-benzoyl-
oxy-2-azido-1-[2-O-(5,5-di-tert-butyloxycarbonylpentyl)-3
-O-benzoyl-4,6-O-benzylidene-ꢀ-D-galactopyranosyloxy]-
4-octadecene 36b. Ethyl 3-O-benzoyl-4,6-O-benzylidene
-2-O-(5,5-di-tert-butyloxycarbonylpentyl)-1-thio-b-d-
galactopyranoside 18 (0.780 g, 1.12 mmol) and
(2S,3R,4E)-2-azido-3-benzoyloxy-4-octadecen-1-ol 28
(0.423 g, 0.99 mmol) were reacted by the procedure used
to synthesize compounds 29a and b and afforded the
a-anomer ot the title compound (0.630 g, 60%) and the
b-anomer of the title compound (ꢀ300 mg, ꢀ29%) as
yellow oils. ꢁ-Anomer: 1H NMR (400 MHz, CDCl3)
d 0.88 (3H, t, J=6.8 Hz, -CH3), 1.25–1.38 (22H, m,
-(CH2)11-), 1.42 (18H, s, 2Â-OtBu), 1.42–1.44, 1.51–1.57
and 1.68–1.74 (3Â2H, 3m, -(CH2)3-), 2.09 (2H, dt,
J=6.8, 6.8 Hz, ¼CH-CH2-), 2.96 (1H, t, J=7.5 Hz,
-CH(CO2tBu)2), 3.55–3.48 (3H, m, H-1 and -OCH2-),
3.87 (1H, dd, J=10.8, 4.2 Hz, H-1), 3.88 (1H, br s, H-
50), 4.03 (1H, m, H-2), 4.09 (1H, dd, J=12.5, 1.2 Hz, H-
60), 4.12 (1H, dd, J=10.5, 3.4 Hz, H-20), 4.29 (1H, dd,
J=12.5, 0.7 Hz, H-60), 4.61 (1H, d, J=3.5 Hz, H-40),
5.18 (1H, d, J=3.4 Hz, H-10), 5.45 (1H, dd, J=10.5,
3.5 Hz, H-30), 5.52 (1H, s, -OCHO-), 5.57–5.66 (2H, m,
H-3 and H-4), 5.97 (1H, dt, J=14.4, 6.8 Hz, H-5), 7.34–
7.36, 7.44–7.49, 7.55–7.60 and 8.06–8.08 (15H, 4 sets of
m, aromatic H). IR (CH2Cl2) ꢀmax (cmÀ1) 2920, 2850
(C–H), 1720 (C¼O). Anal. calcd for C60H83N3O13: C,
68.35; H, 7.93; N, 3.99. Found: C, 68.19; H, 7.79; N,
4.03. ꢀ-Anomer: 1H NMR (400 MHz, CDCl3) d 0.89
(3H, t, J=6.8 Hz, -CH3), 1.12–1.33 (22H, m, -(CH2)11-),
1.38–1.46 (2H, m, -CH2-), 1.41 and 1.42 (18H, 2s,
2Â -OtBu), 1.47–1.60 (2H, m, -CH2-), 1.62–1.70 (2H,
m, -CH2-), 2.09 (2H, dt, J=6.9, 6.9 Hz, ¼CH-CH2-),
2.90 (1H, t, J=7.5 Hz, -CH(CO2tBu)2), 3.54 (1H, s, H-
(5H, 3 sets of m, aromatic H). IR (film) ꢀmax (cmÀ1
)
3290 (NH), 2920, 2850 (C-H), 1740, 1715, 1640 (C¼O).
Anal. calcd for C69H121NO13Si: C, 69.02; H, 10.16; N,
1.17. Found: C, 69.05; H, 10.12; N, 1.23.
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-O-
(3,3-di-carboxypropyl)-ꢁ-D-galactopyranosyloxy]-4-octa-
decene 34a. (2S,3R,4E)-3-Benzoyloxy-2-hexadecanoyl-
amino-1-[2-O-(3,3-di-tert-butyloxycarbonylpropyl)-3,4-O-
isopropylidene-6-O-tert-butyldimethylsilyl-a-d-galacto-
pyranosyloxy]-4-octadecene 32a (0.155 g, 0.129 mmol)
was reacted by the procedure used to synthesize com-
pound 33a and afforded the title compound (0.081 g,
67%) as a beige solid. Diacid: 1H NMR (400 MHz,
pyridine-d5) d 0.85 (6H, t, J=6.7 Hz, 2Â-CH3), 1.23–
1.39 (46H, m, -(CH2)13- and -(CH2)10-), 1.81–1.90 and

A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
1413
50), 3.60–3.69 (2H, m, -OCH2- and H-1), 3.84 (1H, dd,
J=10.1, 7.6 Hz, H-20), 3.82–3.91 (1H, m, -OCH2-),
4.02–4.09 (3H, m, H-60, H-1 and H-2), 4.34 (1H, dd,
J=0.7, 11.9 Hz, H-60), 4.46–4.48 (2H, m, H-10 and H-
40), 5.10 (1H, dd, J=10.1, 3.6 Hz, H-30), 5.51 (1H, s,
-OCHO-), 5.60 (1H, dd, J=15.0, 8.0 Hz, H-4), 5.67
(1H, dd, J=8.0, 3.4 Hz, H-3), 5.96 (1H, dt, J=15.0,
6.9 Hz, H-5), 7.32–7.39, 7.41–7.53, 7.56–7.60 and
H-1, H-30 and H-60), 3.99 (1H, dd, J=10.4, 3.6 Hz, H-
20), 4.34 (1H, br s, H-40), 4.58 (1H, m, H-2), 5.07 (1H, d,
J=3.6 Hz, H-10), 5.38 (1H, dd, J=10.4, 3.1 Hz, H-30),
5.53 (1H, dd, J=15.0, 7.5 Hz, H-4), 5.59 (1H, t, H-3),
5.89 (1H, dt, J=15.0, 6.8 Hz, H-5), 6.18 (1H, d,
J=9.4 Hz, -NH-), 7.44–7.50, 7.56–7.62 and 8.03–8.09
(10H, 3 sets of m, aromatic H). IR (CH2Cl2) ꢀmax
(cmÀ1) 3650–3100 (OH and NH), 2920, 2850 (C–H),
1720, 1650 (C¼O). Anal. calcd for C69H111NO14: C,
70.32; H, 9.49; N, 1.19. Found: C, 70.27; H, 9.42; N, 1.19.
8.05–8.13 (15H,
4 sets of m, aromatic H). IR
(CH2Cl2) ꢀmax (cmÀ1): 3025, 2980, 2930, 2850 (C-H),
2100 (-N3), 1720 (C¼O). Anal. calcd for
C60H83N3O13: C, 68.35; H, 7.93; N, 3.99. Found: C,
68.14; H, 7.92; N, 3.83.
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-O-
(5,5-dicarboxypentyl)-3-O-benzoyl-ꢁ-D-galactopyranosyl-
oxy]-4-octadecene 41a. (2S,3R,4E)-3-Benzoyloxy-2-hexa-
decanoylamino-1-[2-O-(5,5-di-tert-butyloxycarbonyl-
pentyl)-3-O-benzoyl-a-d-galactopyranosyloxy]-4-octa-
decene 40a (0.173 g, 0.147 mmol) was reacted by the
procedure used to synthesize compound 33a and gave
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-O-
(5,5-di-tert-butyloxycarbonylpentyl)-3-O-benzoyl-4,6-O-
benzylidene-ꢁ-D-galactopyranosyloxy]-4-octadecene 37a.
(2S,3R,4E)-3-Benzoyloxy-2-azido-1-[2-O-(5,5-di-tert-
butyloxycarbonylpentyl)-3-O-benzoyl-4,6-O-benzylidene-a-
1
the title compound (0.082 g, 52%) as a white solid. H
d-galactopyranosyloxy]-4-octadecene
36a
(0.620 g,
NMR (400 MHz, pyridine-d5) d 0.85 (6H, t, J=6.6 Hz,
2Â-CH3), 1.24–1.41 (48H, m, -(CH2)11- and -(CH2)13-),
1.68–1.77 (2H, m, -(CH2)-), 1.88, 2.05 and 2.30 (3Â2H,
3m, -(CH2)2- and=CH-CH2-), 2.51 (2H, t, J=7.3 Hz,
-NHCOCH2-), 3.69–3.71 (1H, m, -OCH2-), 3.78 (1H, t,
J=7.4 Hz, -CH(CO2H)2), 3.83–3.87 (1H, m, -OCH2-),
4.26 (1H, dd, J=10.8, 5.9 Hz, H-60), 4.38–4.46 (3H, m,
H-1 and H-60), 4.59 (1H, dd, J=6.0, 6.0 Hz, H-50), 4.64
(1H, dd, J=10.5, 3.4 Hz, H-20), 4.97 (1H, d, J=2.7 Hz,
H-40), 5.21 (1H, m, H-2), 5.57 (1H, d, J=3.4 Hz, H-10),
5.91–5.97 (2H, m, H-30 and H-4), 6.10 (1H, dt, J=15.4,
6.7 Hz, H-5), 6.28 (1H, t, J=6.8 Hz, H-3), 7.38–7.51 and
8.23–8.28 (10H, 2 sets of m, aromatic H), 8.95 (1H, d,
J=8.7 Hz, -NH-). IR (KBr) ꢀmax (cmÀ1): 3700–3100
(OH and NH), 2920, 2850 (C–H), 1720, 1650 (C¼O).
MS (ESI+): 1088.9 (M+Na)+. MS (ESIÀ): 1064.8
(MÀH)À.
0.588 mmol) was reacted by the procedure used to syn-
thesize compound 30a and gave the title compound
1
(0.600 g, 81%) as a beige solid. H NMR (400 MHz,
CDCl3) d 0.89 (6H, t, J=6.6 Hz, 2Â -CH3), 1.23–1.30
(48H, br s, -(CH2)11- and -(CH2)13-), 1.42 (18H, s,
2Â-OtBu), 1.52–1.74 (6H, m, -(CH2)3-), 2.03 (2H, dt,
J=7.0, 7.0 Hz, ¼CH-CH2), 2.22 (2H, m, NHCOCH2-),
2.95 (1H, t, J=7.5 Hz, -CH(CO2tBu)2), 3.54 (1H, dt,
J=9.2, 6.8 Hz, -OCH2-), 3.69 (1H, dt, J=9.2, 6.5 Hz,
-OCH2-), 3.80 (1H, s, H-50), 3.83 (1H, d, J=11.1,
4.1 Hz, H-1), 3.88 (1H, d, J=11.1, 3.4 Hz, H-1), 4.03–
4.09 (2H, m, H-60 and H-20), 4.25 (1H, d, J=12.2 Hz, H-
60), 4.50–4.55 (1H, m, H-2), 4.55 (1H, d, J=3.5 Hz, H-
40), 5.07 (1H, d, J=3.4 Hz, H-10), 5.45 (1H, dd, J=10.5,
3.5 Hz, H-30), 5.50 (1H, s, -OCHO-), 5.53 (1H, dd,
J=15.1, 7.6 Hz, H-4), 5.60 (1H, t, J=7.6 Hz, H-3), 3.88
(1H, dt, J=15.1, 7.0 Hz, H-5), 6.17 (1H, d, J=9.3 Hz,
-NH-), 7.33–7.37, 7.42–7.60 and 8.03–8.09 (15H, 3 sets of
m, aromatic H). IR (CH2Cl2) ꢀmax (cmÀ1): 3300 (NH),
2920, 2850 (C–H), 1720, 1675 (C¼O).
Synthesis of (2S,3R,4E)-3-benzoyloxy-2-hexadecanoyl-
amino-1-[2-O-(5,5-di-carboxypentyl)-3-O-benzoyl-ꢀ-D-
galactopyranosyloxy]-4-octadecene 41b
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-
O-(5,5-di-tert-butyloxycarbonylpentyl)-3-O-benzoyl-4,6-
O-benzylidene-ꢀ-D-galactopyranosyloxy]-4-octadecene
37b. (2S,3R,4E)-3-Benzoyloxy-2-azido-1-[2-O-(5,5-di-
tert-butyloxycarbonylpentyl)-3-O-benzoyl-4,6-O-ben-
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-O-
(5,5-di-tert-butyloxycarbonylpentyl)-3-O-benzoyl-ꢁ-D-ga-
lactopyranosyloxy]-4-octadecene 40a. A stirred solution
of (2S,3R,4E)-3-benzoyloxy-2-hexadecanoylamino-1-[2-
O-(5,5-di-tert-butyloxycarbonylpentyl)-3-O-benzoyl-4,6-O-
benzylidene-a-d-galactopyranosyloxy]-4-octadecene 37a
(0.225 g, 0.178 mmol) in methylene chloride (10 mL) was
treated at 0 ꢁC with aqueous trifluoroacetic acid (90%,
1 mL). The solution was stirred at 0 ꢁC for 3 h, then
solid sodium bicarbonate followed by anhydrous mag-
nesium sulfate were added. The mixture was filtered and
evaporated. The residue was purified by silica gel col-
umn chromatography (20–50% ethyl acetate/hexane)
and afforded the title material (0.158 g, 75%) as an oil.
1H NMR (400 MHz, CDCl3) d 0.89 (6H, t, J=6.7 Hz,
2Â -CH3), 1.24–1.33 (48H, m, -(CH2)11- and -(CH2)13-),
1.43 (18H, 2s, 2Â-OtBu), 1.51–1.74 (6H, m, -(CH2)3-),
1.99–2.06 (2H, m,=CH-CH2-), 2.22 (2H, m, -NH-
COCH2-), 2.50 (1H, t, J=6.2 Hz, -OH-60), 2.63 (1H, d,
J=3.0 Hz, -OH-40), 2.94 (1H, t, J=7.5 Hz, -CH-
(CO2tBu)2), 3.52 (1H, dt, J=9.3, 6.8 Hz, -OCH2-), 3.69
(1H, dt, J=9.3 and 6.4 Hz, -OCH2-), 3.83–3.93 (5H, m,
zylidene-b-d-galactopyranosyloxy]-4-octadecene
36b
(0.371 g, 0.35 mmol) was reacted by the procedure used
to synthesize compound 30a and gave the title com-
pound (0.364 g, 82%) as a beige solid. 1H NMR
(400 MHz, CDCl3) d 0.89 (6H, m, 2Â-CH3), 1.14–1.36
(48H, m, -(CH2)13- and -(CH2)11-), 1.41 and 1.42 (18H,
2 s, 2Â -OtBu), 1.41–1.51 and 1.58–1.67 (6H, 2 sets of
m, -CH2-CH(CO2tBu)2 and -(CH2)2-), 2.03 (2H, q,
J=7.0 Hz,=CH-CH2-), 2.15 (2H, m, -NHCOCH2-),
2.87 (1H, t, J=7.5 Hz, -CH(CO2tBu)2), 3.54 (1H, br s,
H-50), 3.59 (1H, dt, J=9.4, 6.9 Hz, -OCH2-), 3.75–3.86
(3H, m, -OCH2-, H-20 and H-1), 4.06 (1H, d,
J=11.9 Hz, H-60), 4.16 (1H, dd, J=11.2, 4.6 Hz, H-1),
4.28 (1H, d, J=11.9 Hz, H-60), 4.44 (1H, d, J=7.9 Hz,
H-10), 4.46 (1H, d, J=3.8 Hz, H-40), 4.50 (1H, m, H-2),
5.08 (1H, dd, J=10.1, 3.8 Hz, H-30), 5.50 (1H, s,
-OCHO-), 5.53 (1H, dd, J=15.3, 7.4 Hz, H-4), 5.67 (1H,
br t, H-3), 5.87 (1H, dt, J=15.3, 7.0 Hz, H-5), 6.21 (1H,

1414
A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
1
d, J=8.8 Hz, -NH-), 7.34–7.38, 7.42–7.49, 7.54–7.60
and 8.04–8.09 (15H, 4 sets of m, aromatic H). IR
(CH2Cl2) ꢀmax (cmÀ1) 3400 (NH), 3050, 2930, 2850 (C-
H), 1720, 1670 (C¼O). Anal. calcd for C76H115NO14: C,
72.06; H, 9.15; N, 1.11. Found: C, 72.09; H, 9.10; N,
1.29.
Sodium salt: H NMR (400 MHz, CD3OD) d 0.89 (6H,
t, J=6.8 Hz, 2Â -CH3), 1.25–1.31 (46H, m, -(CH2)13-
and -(CH2)10-), 1.40, 1.49, 1.59 and 1.73 (4Â2H, 4 sets
of m, -CH2-, -(CH2)2- and -CH2-CH(CO2Na)2), 2.09
(2H, dt, J=6.9, 6.9 Hz, ¼CH-CH2-), 2.18–2.24 (2H, m,
-NHCOCH2-), 2.97 (1H, br t, -CH(CO2Na)2), 3.56–
3.66 (4H, m, H-50, H-60, H-1 and -OCH2-), 3.69 (1H, dd,
J=10.1, 7.7 Hz, H-20), 3.77–3.83 (1H, m, -OCH2-), 3.84
(1H, dd, J=10.7, 5.4 Hz, H-1), 3.99 (1H, dd, J=10.5,
6.3 Hz, H-60), 4.11 (1H, d, J=3.1 Hz, H-40), 4.43 (1H, d,
J=7.7 Hz, H-10), 4.51 (1H, m, H-2), 4.94 (1H, dd,
J=10.1, 3.3 Hz, H-30), 5.56 (1H, dd, J=15.2, 7.7 Hz, H-
4), 5.65 (1H, br t, H-3), 5.91 (1H, dt, J=15.2, 6.9 Hz, H-
5), 7.45–7.51, 7.58–7.61 and 7.99–8.08 (10H, 3 sets of m
aromatic H). IR (nujol) ꢀmax (cmÀ1) 3650–3100 (NH
and OH), 2920, 2850 (C–H), 1720, 1650, 1595 (C¼O).
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-
O-(5,5-di-tert-butyloxycarbonylpentyl)-3-O-benzoyl-ꢀ-D-
galactopyranosyloxy]-4-octadecene 40b. (2S,3R,4E)-3-
Benzoyloxy-2-hexadecanoylamino-1-[2-O-(5,5-di-tert-
butyloxycarbonylpentyl)-3-O-benzoyl-4,6-O-benzylidene-b-
d-galactopyranosyloxy]-4-octadecene
37b
(0.436 g,
0.27 mmol) was reacted by the procedure used to syn-
thesize compound 40a and afforded the title compound
1
(0.259 g, 81%) as a white solid. H NMR (400 MHz,
CDCl3) d 0.89 (6H, t, J=6.8 Hz, 2Â-CH3), 1.19–1.39
(48H, m, -(CH2)13- and -(CH2)11-), 1.43 (18H, s, 2Â
-OtBu), 1.46–1.53 and 1.58–1.73 (2H and 4H, -CH2-
CH(CO2tBu)2 and -(CH2)2-), 2.03–2.25 (2H, br s, 2Â
-OH), 2.05 (2H, dt, J=6.9, 6.9 Hz, ¼CH-CH2-), 2.19
(2H, m, -NHCOCH2-), 2.89 (1H, t, J=7.5 Hz, -CH-
(CO2tBu)2), 3.43 (1H, br dd, J=3.4, 3.4 Hz, H-50), 3.60
(1H, dt, J=9.4, 6.6 Hz, -OCH2-), 3.67 (1H, dd, J=12.5,
3.2 Hz, H-60), 3.75–3.85 (4H, m, H-20, H-1, H-60 and
-OCH2-), 4.01 (1H, dd, J=10.5, 2.7 Hz, H-1), 4.27 (1H,
d, J=2.9 Hz, H-40), 4.44 (1H, d, J=7.6 Hz, H-10), 4.62
(1H, m, H-2), 4.98 (1H, dd, J=10.0, 3.0 Hz, H-30), 5.55
(1H, dd, J=15.4, 7.9 Hz, H-4), 5.77 (1H, t, J=7.9 Hz,
H-3), 5.86 (1H, d, J=9.4 Hz, -NH-), 5.95 (1H, dt,
J=15.4, 6.9 Hz, H-5), 7.46–7.49, 7.57–7.62 and 8.05–
8.10 (10H, 3 sets of m, aromatic H). IR (CH2Cl2) ꢀmax
(cmÀ1) 3600, 3450–3350 (NH and OH), 3050, 2930,
2850 (C–H), 1720, 1675 (C¼O). Anal. calcd for
C69H111NO14: C, 70.32; H, 9.49; N, 1.19. Found: C,
70.78; H, 9.52; N, 1.30.
Synthesis of (2S,3R,4E)-3-benzoyloxy-2-hexadecanoyl-
amino-1-[2-O-(3,3-di-carboxypropyl)-3-O-benzoyl-ꢁ-D-
galactopyranosyloxy]-4-octadecene 43a
(2S,3R,4E)-3-Benzoyloxy-2-azido-1-[2-O-(3,3-di-tert-
butyloxycarbonylpropyl)-3-O-benzoyl-4,6-O-benzylidene-
ꢁ-D-galactopyranosyloxy]-4-octadecene 38a and (2S,
3R,4E)-3-benzoyloxy-2-azido-1-[2-O-(3,3-di-tert-butyl-
oxycarbonylpropyl)-3-O-benzoyl-4,6-O-benzylidene-ꢀ-D-
galactopyranosyloxy]-4-octadecene 38b. Ethyl 2-O-(3,3-
di-tert-butyloxycarbonylpropyl)-3-O-benzoyl-4,6-O-ben-
zylidene-1-thio-b-d-galactopyranoside (1.97g, 3.00 mmol)
20, 2,6-di-tert-butyl-4-methylpyridine (1.23 g, 6.0 mmol)
and (2S,3R,4E)-2-azido-3-benzoyloxy-4-octadecen-1-ol
28 were reacted by the procedure used to synthesize
compounds 29a and b except that dioxane was used as
solvent. This gave the a-anomer (1.64 g, 64%) and the
b-anomer (0.51 g, 20%) of the title compound as oils.
ꢁ-anomer: [a]2D2 +71.8ꢁ (c 1.2, CHCl3). 1H NMR
(400 MHz, CDCl3) d 0.89 (3H, t, J=6.8 Hz, -CH3), 1.26
(20H, br s, -(CH2)10-), 1.39–1.42 (20H, m, 2Â-OtBu and
-CH2-), 2.01–2.12 (4H, m, ¼CH-CH2- and -CH2–
CH(CO2tBu)2), 3.41 (1H, t, J=7.4 Hz, -CH(CO2tBu)2),
3.64–3.72 (3H, m, H-2, H-20 and -OCH2-), 3.89 (1H, dd,
J=10.7, 4.2 Hz, H-1), 3.90 (1H, br s, H-50), 4.04 (1H, dt,
J=7.8, 4.0 Hz, -OCH2-), 4.10 (1H, br d, J=12.5 Hz, H-
60), 4.14 (1H, dd overlapped by H-60, H-1), 4.30 (1H, br
d, J=12.5 Hz, H-60), 4.64 (1H, d, J=3.4 Hz, H-40), 5.19
(1H, d, J=3.4 Hz, H-10), 5.45 (1H, dd, J=10.5, 3.4 Hz,
H-30), 5.52 (1H, s, -OCHO-), 5.62 (1H, dd, J=14.3,
8.0 Hz, H-4), 5.66 (1H, dd, J=8.0, 4.1 Hz, H-3), 5.98
(1H, dt, J=14.3, 6.8 Hz, H-5), 7.33–7.34, 7.43–7.49,
7.56–7.60 and 8.07–8.13 (15H, 4 sets of m, 3Â-C6H5).
IR (film) ꢀmax (cmÀ1) 3035, 3010, 2970, 2925, 2855 (C–
H), 2100 (N3), 1722 (C¼O). Anal. calcd for
C58H79N3O13: C, 67.88; H, 7.76; N, 4.09. Found: C,
67.83; H, 7.64; N, 4.20. ꢀ-Anomer: [a]2D2 +28ꢁ (c 0.97,
CHCl3). 1H NMR (400 MHz, CDCl3) d 0.89 (3H, t,
J=6.8 Hz, -CH3), 1.20–1.50 (22H, m, -(CH2)11-), 1.35
and 1.36 (18H, 2 s, 2Â-OtBu), 1.96–2.11 (4H, m, ¼CH–
CH2- and -CH2-CH(CO2tBu)2), 3.28 (1H, t, J=7.3 Hz,
-CH(CO2tBu)2), 3.54 (1H, br s, H-50), 3.69 (1H, dt,
J=9.9, 6.6 Hz, -OCH2-), 3.72 (1H, dd, J=10.2, 4.6 Hz,
H-1), 3.86 (1H, dd, J=10.1, 7.7 Hz, H-20), 3.94 (1H, dt,
J=9.9, 6.0 Hz, -OCH2-), 4.02 (1H, dd, J=10.2, 7.9 Hz,
H-1), 4.07 (1H, d, J=12.2 Hz, H-60), 4.10–4.13 (1H, m,
H-2), 4.34 (1H, d, J=12.2 Hz, H-60), 4.47–4.50 (2H, m,
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-
O-(5,5-di-carboxypentyl)-3-O-benzoyl-b-D-galactopyr-
anosyloxy]-4-octadecene 41b. (2S,3R,4E)-3-Benzoyloxy-
2-hexadecanoylamino-1-[2-O-(5,5-di-tert-butyloxycarb-
onylpentyl)-3-O-benzoyl-b-d-galactopyranosyloxy]-4-
octadecene (0.240 g, 0.2 mmol) 40b was reacted by the
procedure used to synthesize compound 33a and gave
the title compound (0.139 g, 65%) as a white solid.
1
Diacid: H NMR (400 MHz, pyridine-d5) d 0.85 (6H,
t, J=6.7 Hz, 2Â-CH3), 1.24 (44H, br s, -(CH2)12- and
-(CH2)10-), 1.41, 1.67–1.92, 2.04 and 2.24–2.32 (2H,
6H, 2Â2H, 4 sets of m, 2Â-CH2-, -(CH2)2-, -CH2-
CH(CO2H)2 and ¼CH-CH2-), 2.49 (2H, t, J=7.4 Hz,
-NHCOCH2-), 3.73 (1H, t, J=7.4 Hz, -CH(CO2H)2),
3.90 (1H, m, -OCH2-), 4.07 (1H, dd, J=6.2, 6.2 Hz, H-
50), 4.19 (1H, m, -OCH2-), 4.27 (1H, dd, J=10.4, 5.2 Hz,
H-1), 4.32 (1H, dd, J=10.9, 6.1 Hz, H-60), 4.37–4.43
(2H, m, H-20 and H-60), 4.52 (1H, dd, J=10.4, 6.7 Hz,
H-1), 4.91–4.93 (2H, m, H-10 and H-40), 5.25 (1H, m, H-
2), 5.57 (1H, dd, J=10.1, 3.1 Hz, H-30), 5.94 (1H, dd,
J=15.4, 7.3 Hz, H-4), 6.12 (1H, dt, J=15.4, 6.7 Hz, H-
5), 6.28 (1H, br t, H-3), 7.19–7.50 and 8.24–8.26 (10H, 2
sets of m, aromatic H), 8.60 (1H, d, J=8.7 Hz, -NH-).
IR (nujol) ꢀmax (cmÀ1) 3700–3100 (NH and OH), 2930,
2850 (C-H), 1715, 1645 (C¼O). MS (ESIÀ): 1064.7
(MÀH)À. MS (ESI+): 1066.7 (MH+), 1088.7 (M+Na)+.

A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
1415
H-10 and H-40), 5.07 (1H, dd, J=10.1, 3.6 Hz, H-30),
5.50 (1H, s, -OCHO-), 5.60 (1H, dd, J=14.4, 8.0 Hz, H-
4), 5.64 (1H, dd, J=8.0, 4.0 Hz, H-3), 5.95 (1H, dt,
J=14.4, 6.7 Hz, H-5), 7.33–7.37, 7.43–7.49, 7.53–7.59
and 8.07–8.13 (15H, 4 sets of m, aromatic H). IR (film)
ꢀmax (cmÀ1) 2930, 2850 (C-H), 2100 (-N3), 1725 (C¼O).
Anal. calcd for C58H79N3O13. CH2Cl2: C, 63.77; H,
7.35; N, 3.78. Found: C, 64.00; H, 7.35; N, 3.89.
ꢀmax (cmÀ1) 3600–3100 (OH and NH), 2920, 2850 (C–
H), 1720, 1650 (C¼O).
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-
O-(3,3-di-carboxypropyl)-3-O-benzoyl-ꢁ-D-galactopyr-
anosyloxy]-4-octadecene 43a. (2S,3R,4E)-3-Benzoyloxy-
2-hexadecanoylamino-1-[2-O-(3,3-di-tert-butyloxycar-
bonylpropyl)-3-O-benzoyl-a-d-galactopyranosyloxy]-4-
octadecene 42a (0.083 g, 0.072 mmol) was reacted by the
procedure used to synthesize compound 33a and affor-
ded the title compound (0.074 g, 98%) as a white fluffy
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-
O-(3,3-di-tert-butyloxycarbonylpropyl)-3-O-benzoyl-4,6-
O-benzylidene-ꢁ-D-galactopyranosyloxy]-4-octadecene
39a. (2S,3R,4E)-3-Benzoyloxy-2-azido-1-[2-O-(3,3-di-
tert-butytloxycarbonylpropyl)-3-O-benzoyl-4,6-O-ben-
solid. [a]2D2 +66ꢁ (c 0.5, CHCl3). H NMR (400 MHz,
1
DMSO-d6) d 0.85 (6H, t, J=6.8 Hz, 2Â-CH3), 1.19–1.35
(46H, m, -(CH2)11- and -(CH2)12-), 1.47, 1.85, 2.00 and
2.09 (8H, 4 sets of m, -CH2-, -NHCOCH2-, ¼CH-CH2-
and -CH2CH(CO2H)2), 3.42 (1H, dd, J=10.9 and
6.5 Hz, H-1), 3.47–3.49, 3.51–3.58 and 3.73–3.76 (7H, 3
sets of m, H-50, H-60, H-1, -OCH2- and -CH(CO2H)2),
3.85 (1H, dd, J=10.5, 3.4 Hz, H-20), 4.10 (1H, br s, H-
40), 4.37 (1H, m, H-2), 4.61 (1H, br s, -OH-60), 4.97 (1H,
d, J=3.4 Hz, H-10), 5.06 (1H, dd, J=10.5, 3.0 Hz, H-30),
5.09 (1H, s, -OH-40), 5.48 (1H, br t, H-3), 5.55 (1H, dd,
J=15.1, 7.5 Hz, H-4), 5.7 (1H, dt, J=15.1, 6.8 Hz, H-5),
7.47–7.52, 7.61–7.67 and 7.94–8.01 (11H, 3 sets of m,
2Â -C6H5 and -NH-), 12.63 (1H, br s, -COOH). IR
(film) ꢀmax (cmÀ1) 3700–3000 (OH and NH), 2920, 2850
(C-H), 1720, 1650 (C¼O). MS (ESIÀ) 1036.6 (MÀH)À.
MS (ESI+) 1038.7 (MH+), 1060.7 (M+Na)+. HRMS
(Pos. FAB): calcd for C59H91O14NNa: 1060.634711.
Found: 1060.63460 d=À1.0 ppm.
zylidene-a-d-galactopyranosyloxy]-4-octadecene
38a
(0.477 g, 0.464 mmol) was reacted by the procedure used
to synthesize compound 30a and afforded the title com-
pound (0.527 g, 92%) as an oil. [a]2D2 +61.5ꢁ (c 1.1,
CHCl3). 1H NMR (400MHz, CDCl3) d 0.87–0.91 (6H, m,
2Â -CH3), 1.24–1.44 (44H, m, -(CH2)10- and -(CH2)12-),
1.37 and 1.39 (18H, 2s, 2Â-OtBu), 1.53–1.65 (4H, m, 2Â
-CH2-), 2.00–2.09 and 2.19–2.40 (6H, 2 sets of m,
-NHCOCH2-, ¼CH-CH2- and -CH2-CH(CO2tBu)2),
3.39 (1H, t, J=7.3 Hz, -CH(CO2tBu)2), 3.62 (1H, dt,
J=9.5, 5.8 Hz, -OCH2-), 3.72–3.82 (2H, m, -OCH2- and
H-1), 3.80 (1H, br s, H-50), 3.91 (1H, dd, J=10.9,
3.4 Hz, H-1), 4.05 (1H, dd, J=12.4, 1.3 Hz, H-60), 4.08
(1H, dd, J=10.6, 3.4 Hz, H-20), 4.25 (1H, d, J=12.4 Hz,
H-60), 4.52 (1H, m, H-2), 4.59 (1H, d, J=3.4 Hz, H-40),
5.08 (1H, d, J=3.4 Hz, H-10), 5.39 (1H, dd, J=10.6,
3.4 Hz, H-30), 5.50 (1H, s, -OCHO-), 5.55 (1H, dd,
J=15.3, 7.5 Hz, H-4), 5.67 (1H, t, J=7.3 Hz, H-3), 5.92
(1H, dt, J=15.3, 6.8 Hz, H-5), 6.44 (1H, d,
J=9.0 Hz, -NH-), 7.31–7.33, 7.42–7.48, 7.54–7.60,
8.04–8.12 (15H, 4 sets of m, 3Â-C6H5). IR (film) ꢀmax
(cmÀ1): 3500–3240 (NH), 2925, 2855 (C–H), 1722, 1650
(C¼O). Anal. calcd for C74H111NO14: C, 71.75; H, 9.03;
N, 1.13. Found: C, 72.02; H, 9.12; N, 1.33.
Synthesis of (2S,3R,4E)-3-benzoyloxy-2-hexadecanoyla-
mino-1-[2-O-(3,3-di-carboxypropyl)-3-O-benzoyl-ꢀ-D-ga-
lactopyranosyloxy]-4-octadecene 43b
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-
O-(3,3-di-tert-butyloxycarbonylpropyl)-3-O-benzoyl-4,6-
O-benzylidene-ꢀ-D-galactopyranosyloxy]-4-octadecene
39b. (2S,3R,4E)-3-Benzoyloxy-2-azido-1-[2-O-(3,3-di-
tert-butyloxycarbonylpropyl)-3-O-benzoyl-4,6-O-benzyli-
dene-b-d-galactopyranosyloxy]-4-octadecene 38b (0.467 g,
0.455 mmol) was reacted by the procedure used to
synthesize compound 30a and afforded the title com-
pound (0.393 g, 70%) as a clear sticky oil. [a]2D2 +36ꢁ (c
1.3, CHCl3). 1H NMR (400 MHz, CDCl3) d 0.90 (6H, t,
J=6.7 Hz, 2Â-CH3), 1.24–1.36 (46H, m, -(CH2)13- and
-(CH2)10-), 1.39 and 1.41 (18H, 2 s , 2Â-OtBu), 1.53–
1.60 (2H, m, -CH2-), 2.00–2.12 (4H, m, -CH2–
CH(CO2tBu)2 and ¼CH-CH2-), 2.21 (2H, t, J=7.4 Hz,
-NHCOCH2-), 3.26 (1H, t, J=7.2 Hz, -CH(CO2tBu)2),
3.50 (1H, br s, H-50), 3.70 (1H, dt, J=9.6, 5.9 Hz,
-OCH2-), 3.78 (1H, dd, J=10.1, 7.7 Hz, H-20), 3.76–3.83
(1H, m overlapped by H-20, -OCH2-), 3.86 (1H, dd,
J=11.1, 3.5 Hz, H-1), 4.00 (1H, d, J=12.2 Hz, H-60),
4.16–4.20 (2H, m, H-1 and H-60), 4.41 (1H, d,
J=7.7 Hz, H-10), 4.45 (1H, d, J=3.6 Hz, H-40), 4.52
(1H, m, H-2), 5.04 (1H, dd, J=10.1, 3.6 Hz, H-30), 5.45
(1H, s, -OCHO-), 5.54 (1H, dd, J=15.5, 7.5 Hz, H-4),
5.69 (1H, t, J=7.5 Hz, H-3), 5.92 (1H, dt, J=15.5,
6.7 Hz, H-5), 6.72 (1H, d, J=8.7 Hz, -NH-), 7.32–7.60
and 8.04–8.09 (15H, 2 sets of m, aromatic H). IR (film)
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-
O-(3,3-di-tert-butyloxycarbonylpropyl)-3-O-benzoyl-ꢁ-D-
galactopyranosyloxy]-4-octadecene 42a. (2S,3R,4E)-3-
Benzoyloxy-2-hexadecanoylamino-1-[2-O-(3,3-di-tert-
butyloxycarbonylpropyl-3-O-benzoyl-4,6-O-benzylidene-
a-d-galactopyranosyloxy]-4-octadecene 39a (0.242 g,
0.195 mmol) was reacted by the procedure used to syn-
thesize compound 40a and afforded the title material
(0.187 g, 83%) as an oil. [a]2D2 +60.1ꢁ (c 1.0, CHCl3).
1H NMR (400 MHz, CDCl3) d 0.89 (6H, t, J=6.7 Hz,
2Â -CH3), 1.24–1.47 (62H, m, -(CH2)10-, -(CH2)12- and
2Â-OtBu), 1.63 (5H, m, 2Â-CH2- and -OH), 2.00–2.08
(4H, m, ¼CH–CH2- and -CH2–CH(CO2tBu)2), 2.22
(2H, m, -NHCOCH2-), 2.65 (1H, br s, -OH), 3.35 (1H,
t, J=7.4 Hz, -CH(CO2tBu)2), 3.61 (1H, dt, J=9.5,
6.0 Hz, -OCH2-), 3.73 (1H, dt, J=9.5, 6.2 Hz, -OCH2-),
3.82–3.94 (4H, m, H-1 and H-60), 3.92 (1H, br s, H-50),
4.00 (1H, dd, J=10.5, 3.6 Hz, H-20), 4.37 (1H, d,
J=2.9 Hz, H-40), 4.56 (1H, m, H-2), 5.07 (1H, d,
J=3.6 Hz, H-10), 5.32 (1H, dd, J=10.5 and 3.0 Hz, H-
30), 5.54 (1H, dd, J=15.3, 7.4 Hz, H-4), 5.64 (1H, t,
J=7.2 Hz, H-3), 5.91 (1H, dt, J=15.3, 6.8 Hz, H-5),
6.38 (1H, d, J=9.1 Hz, -NH-), 7.43–7.49, 7.55–7.61
and 8.04–8.11 (10H, 3 sets of m, 3Â-C6H5). IR (film)
ꢀ
max (cmÀ1): 2920, 2850 (C-H), 1725, 1680 (C¼O). Anal.
calcd for C74H111NO14: C, 71.75; H, 9.03; N, 1.13.
Found: C, 71.77; H, 9.09; N, 1.23.

1416
A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[2-
3-benzoyloxy-4-octadecen-1-ol 28 (0.540 g, 1.68 mmol)
were reacted by the procedure used to synthesize com-
pounds 29a and b except that acetonitrile was used as
solvent. This afforded the a-anomer (0.694 g, 39%) and
the b-anomer (0.453 g, 25%) of the title compound as
O-(3,3-di-carboxypropyl)-3-O-benzoyl-b-D-galactopyr-
anosyloxy]-4-octadecene 43b. (2S,3R,4E)-3-Benzoyloxy-
2-hexadecanoylamino-1-[2-O-(3,3-di-tert-butyloxycar-
bonylpropyl)-3-O-benzoyl-4,6-O-benzylidene-b-d-galac-
topyranosyloxy]-4-octadecene 39b (0.350 g, 0.28 mmol)
was treated with anisole (65 mL, 0.56 mmol) followed by
90% aqueous trifluoroacetic acid (6 mL) at 5 ꢁC. This
mixture was stirred for 5 min at 5 ꢁC and then the ice-
bath was removed and this was stirred for 10 min.
Toluene was added and the mixture was evaporated
under vaccuo and coevaporated with toluene (2Â). This
was repeated until there was no starting material left.
The residue was dried in vaccuo, dissolved in chloro-
form and filtered on Millex LCR carthridge. The solu-
tion was triturated with acetonitrile. The residue was
recrystallized form chloroform/acetonitrile to give the
title compound (0.120 g, 41%) as a white solid.
1
yellow oils. ꢁ-Anomer: H NMR (400 MHz, CDCl3) d
0.89 (3H, t, J=6.8 Hz, -CH3), 1.26 (20H, m, -(CH2)10-),
1.36–1.59 (4H, m, -(CH2)2-), 1.45 and 1.45 (18H, 2 s,
2Â-OtBu), 1.67 and 1.83 (2Â2H, 2 sets of m, -(CH2)2-),
2.08 (2H, q, J=6.9 Hz,=CH-CH2-), 3.10 (1H, t,
J=7.5 Hz, -CH(CO2tBu)2), 3.56–3.76 (5H, m, -OCH2-,
H-1 and H-2), 3.79 (3H, s, -OCH3), 3.83 (1H, dd,
J=10.2, 3.1 Hz, H-30), 3.96 (1H, dd, J=10.2, 3.5 Hz,
H-20), 3.94–3.99 (1H, m overlapped by H-20, H-50), 4.07
(1H, dd, J=12.3, 1.2 Hz, H-60), 4.23 (1H, d, J=12.3 Hz,
H-60), 4.33 (1H, d, J=3.1 Hz, H-40), 4.56 (1H, d,
J=11.6 Hz, -OCH2Ar), 4.75 (1H, d, J=11.6 Hz,
-OCH2Ar), 4.85 (1H, d, J=3.5 Hz, H-10), 5.55 (1H, s,
-OCHO-), 5.55–5.64 (1H, m overlapped by H-3, H-4),
5.63 (1H, dd, J=7.9, 4.0 Hz, H-3), 5.93 (1H, dt,
J=14.5, 6.9 Hz, H-5), 6.82–6.85, 7.17–7.39, 7.45–
7.53, 7.58–7.62 and 8.06–8.08 (14H, 5 sets of m,
aromatic H). IR (CH2Cl2) ꢀmax (cmÀ1) 3050, 2980, 2930
and 2860 (C-H), 1720 (C¼O). Anal. calcd for
C61H87N3O13: C, 68.45; H, 8.19; N, 3.93. Found: C,
68.46; H, 8.03; N, 3.94. ꢀ-anomer: 1H NMR
(400 MHz, CDCl3) d 0.89 (3H, t, J=6.8 Hz, -CH3),
1.24–1.42 (20H, m, -(CH2)10-), 1.42–1.61 (4H, m,
-(CH2)2-), 1.45 and 1.45 (18H, 2 s, 2Â-OtBu), 1.67 and
1.82 (2Â2H, 2 sets of m, -(CH2)2-), 2.04 (2H, m, ¼CH–
CH2-), 3.09 (1H, t, J=7.5 Hz, -CH(CO2tBu)2), 3.38 (1H,
br s, H-50), 3.43 (1H, dd, J=9.7, 3.5 Hz, H-30), 3.57–
3.63 (2H, m, -OCH2- and H-2), 3.69 (1H, dt,
J=9.1, 6.7 Hz, -OCH2-), 3.77 (1H, dd, J=9.6, 7.7 Hz,
H-20), 3.79 (3H, s, -OCH3), 3.99–4.08 (3H, m, H-1 and
H-60), 4.25 (1H, d, J=3.5 Hz, H-40), 4.32 (1H, dd,
J=12.3, 0.9 Hz, H-60), 4.40 (1H, d, J=7.7 Hz, H-10),
4.73 (1H, d, J=10.4 Hz, -OCH2Ar), 4.82 (1H, d,
J=10.4 Hz, -OCH2Ar), 5.55 (1H, s, -OCHO-),
5.58 (1H, dd, J=15.3, 7.9 Hz, H-4), 5.71 (1H, dd,
J=8.0, 3.6 Hz, H-3), 5.90 (1H, dt, J=15.3, 6.8 Hz, H-5),
6.82–6.90, 7.01–7.60 and 8.07–8.10 (14H, 3 sets of m,
aromatic H). IR (CH2Cl2) ꢀmax (cmÀ1) 3050, 2980, 2930
and 2850 (C–H), 1720 (C¼O). Anal. calcd for
C61H87N3O13: C, 68.45; H, 8.19; N, 3.93. Found: C,
68.57; H, 8.03; N, 3.92.
1
Diacid: H NMR (400 MHz, pyridine-d5) d 0.85 (6H, t,
J=6.7 Hz, 2Â-CH3), 1.23–1.41 (46H, m, -(CH2)13-
and -(CH2)11-), 1.87 (2H, m, -CH2-), 2.03 (2H, m, -CH2–
CH(CO2H)2), 2.54 (2H, t, J=7.3 Hz, -NHCOCH2-),
2.76 (2H, m, ¼CH-CH2-), 4.06 (1H, br t, H-50), 4.20
(1H, t, J=7.2 Hz, -CH(CO2H)2), 4.26 (1H, dd, J=10.5,
5.7 Hz, H-60), 4.29–4.39 and 4.47–4.57 (3H, m, -OCH2-
and H-1), 4.36 (1H, dd, J=10.7, 6.7 Hz, H-1), 4.49
(1H, dd, J=10.0, 7.7 Hz, H-20), 4.55 (1H, dd,
J=10.5, 6.7 Hz, H-60), 4.93–4.96 (2H, m, H-10 and
H-40), 5.25 (1H, m, H-2), 5.55 (1H, dd, J=10.1, 3.1 Hz,
H-30), 5.94 (1H, dd, J=15.4, 7.1 Hz, H-4), 6.14 (1H, dt,
J=15.4, 6.7 Hz, H-5), 6.29 (1H, br t, H-3), 7.22–
7.40, 7.45–7.49 and 8.23–8.28 (10H, m, aromatic H),
8.59 (1H, d, J=8.6 Hz, -NH-). Anal. calcd for
C59H91NO14.H2O: C, 67.08; H, 8.87; N, 1.33. Found: C,
67.02; H, 8.75; N, 1.83. Sodium salt: 1H NMR (400 MHz,
CD3OD) d 0.82 (6H, t, J=6.7 Hz, 2Â-CH3), 1.19–1.32
(46H, m, -(CH2)13- and -(CH2)10-), 1.52 (2H, m, -CH2-),
2.00 and 2.20 (4H and 2H, 2 sets of m, -NHCOCH2-,
¼CH-CH2- and -CH2-CH(CO2Na)2), 3.04 (1H, t,
J=7.0 Hz, -CH(CO2Na)2), 3.45–3.51, 3.63–3.70 and
3.76–3.82 (3H, 2H and 1H, 3 sets of m, -OCH2- , H-20,
H-50, H-60 and H-1), 3.77 (1H, dd, J=10.7, 4.1 Hz, H-
1), 3.99 (1H, dd, J=10.4, 6.2 Hz, H-60), 4.04 (1H, br d,
H-40), 4.34 (1H, d, J=7.6 Hz, H-10), 4.41 (1H, m, H-2),
4.82 (1H, dd, J=10.2, 3.4 Hz, H-30), 5.50 (1H, dd,
J=15.3, 7.2 Hz, H-4), 5.61 (1H, t, J=7.2 Hz, H-3), 5.85
(1H, dt, J=15.3, 7.6 Hz, H-5), 7.39–7.43, 7.49–7.54 and
7.95–8.02 (10H, m, aromatic H). IR (KBr) ꢀmax (cmÀ1
)
3700–3100 (O–H), 2920, 2850 (C–H), 1715, 1650, 1600
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[3-
O-(5,5-di-tert-butyloxycarbonylpentyl)-2-O-p-methoxy-
benzyl-4,6-O-benzylidene-ꢁ-D-galactopyranosyloxy]-4-
octadecene 45a. (2S,3R,4E)-3-Benzoyloxy-2-azido-1-[3-
O-(5,5-di-tert-butyloxycarbonylpentyl)-2-O-p-methoxy-
benzyl-4,6-O-benzylidene-a-d-galactopyranosyloxy]-4-
octadecene 44a (0.718 g, 0.67 mmol) was reacted by the
procedure used to synthesize compound 30a and affor-
ded the title compound (0.710 g, 83%) as a white solid.
1H NMR (400 MHz, CDCl3) d 0.89 (6H, t, J=6.7 Hz,
2Â-CH3), 1.24–1.27 (46H, m, -(CH2)13- and -(CH2)10-),
1.42–1.45 (2H, m, -CH2-), 1.45 (18H, 2 s, 2Â-OtBu), 1.5–
1.62 (2H, m, -CH2-), 1.68, 1.85, 2.00 and 2.09 (4Â2H, 4
sets of m, -NHCOCH2-, ¼CH–CH2- and -(CH2)2-), 3.10
(1H, t, J=7.5 Hz, -CH(CO2tBu)2), 3.57 (1H, dt, J=9.0,
6.7 Hz, -OCH2-), 3.64–3.69 (1H, m, -OCH2-), 3.69 (1H,
(C¼O).
Synthesis of (2S,3R,4E)-3-benzoyloxy-2-hexadecanoyla-
mino-1-[3-O-(5,5-di-carboxypentyl)-2-O-benzoyl-ꢁ-D-ga-
lactopyranosyloxy]-4-octadecene 52a
(2S,3R,4E)-3-Benzoyloxy-2-azido-1-[3-O-(5,5-di-tert-
butyloxycarbonylpentyl)-2-O-p-methoxybenzyl-4,6-O-
benzylidene-ꢁ-D-galactopyranosyloxy]-4-octadecene 44a
and (2S,3R,4E)-3-benzoyloxy-2-azido-1-[3-O-(5,5-di-tert-
butyloxycarbonylpentyl)-2-O-p-methoxybenzyl-4,6-O-
benzylidene-ꢀ-D-galactopyranosyloxy]-4-octadecene 44b.
Ethyl 2-O-p-methoxybenzyl-4,6-di-O-benzylidene-3-(5,5-
di-tert-butyloxycarbonylpentyl)-1-thio-b-d-galactopyr-
anoside 26 (1.185 g, 1.68 mmol) and (2S,3R,4E)-2-azido-

A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
1417
br s, H-50), 3.76–3.83 (3H, m, H-1 and H-20), 3.78 (3H,
s, -OCH3), 3.95 (1H, dd, J=10.1, 3.5 Hz, H-30), 4.05
(1H, d, J=11.7 Hz, H-60), 4.21 (1H, d, J=11.7 Hz, H-
60), 4.31 (1H, br d, H-40), 4.48 (1H, m, H-2), 4.62 (1H, d,
J=11.1 Hz, -OCH2Ar), 4.72 (1H, d, J=11.1 Hz,
-OCH2Ar), 4.88 (1H, d, J=3.5 Hz, H-10), 5.48 (1H, dd,
J=15.3, 7.6 Hz, H-4), 5.54 (1H, s, -OCHO-), 5.59 (1H,
t, J=7.4 Hz, H-3), 5.79 (1H, dt, J=15.3, 6.7 Hz, H-5),
5.99 (1H, d, J=9.1 Hz, -NH-), 6.81–6.83, 7.30–7.38,
7.45–7.53, 7.57–7.61 and 8.04–8.06 (14H, 5 sets of m,
aromatic H). IR (CH2Cl2) ꢀmax (cmÀ1): 3440 (NH),
3050, 2990, 2930 and 2860 (C–H), 1720, 1670 (C¼O).
Anal. calcd for C77H119NO14: C, 72.10; H, 9.35; N, 1.09.
Found: C, 72.19; H, 9.31; N, 1.23.
0.22 mmol) was reacted by the procedure used to syn-
thesize compound 18 and afforded the title compound
(0.262 g, 95%) as a white gummy solid. 1H NMR
(400 MHz, CDCl3) d 0.89 (6H, m, 2Â-CH3), 1.25–1.38
(48H, m, -(CH2)13- and -(CH2)11-), 1.42 and 1.43 (18H,
2 s, 2Â-OtBu), 1.54–1.63 (4H, m, -(CH2)2-), 1.74 (2H, m,
-CH2-CH(CO2tBu)2), 2.02 (4H, m, -NHCOCH2- and
¼CH-CH2-), 2.97 (1H, t, J=7.5 Hz, -CH(CO2tBu)2),
3.55 (1H, dt, J=9.2, 6.6 Hz, -OCH2-), 3.65–3.70 (2H, m,
-OCH2- and H-1), 3.81 (1H, s, H-50), 3.86 (1H, dd,
J=11.0, 4.0 Hz, H-1), 3.99 (1H, dd, J=10.5, 3.4 Hz, H-
30), 4.11 (1H, d, J=12.5 Hz, H-60), 4.30 (1H, dd,
J=12.5, 0.9 Hz, H-60), 4.41 (1H, br d, H-40), 4.53 (1H,
m, H-2), 5.33 (1H, d, J=3.7 Hz, H-10), 5.45 (1H, dd,
J=10.4, 7.4 Hz, H-20), 5.48 (1H, dd, J=7.3, 14.7 Hz, H-
4), 5.53 (1H, t, J=7.3 Hz, H-3), 5.60 (1H, s, -OCHO-),
5.64 (1H, d, J=9.3 Hz, -NH-), 5.78 (1H, dt, J=14.7,
6.7 Hz, H-5), 7.32–7.46, 7.54–7.60 and 7.97–8.04 (15H, 3
sets of m, aromatic H). IR (CH2Cl2) ꢀmax (cmÀ1): 3440
(NH), 3050, 2980, 2930 and 2850 (C–H), 1720, 1675
(C¼O). Anal. calcd for C76H115NO14: C, 72.06; H, 9.15;
N, 1.11. Found: C, 72.13; H, 9.11; N, 1.29.
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[3-
O-(5,5-di-tert-butyloxycarbonylpentyl)-4,6-O-benzyli-
dene-a-D-galactopyranosyloxy]-4-octadecene 48a. A solu-
tion of (2S,3R,4E)-3-benzoyloxy-2-hexadecanoylamino-
1-[3-O-(5,5-di-tert-butyloxycarbonylpentyl)-2-O-p-meth-
oxybenzyl-4,6-O-benzylidene-a-d-galactopyranosyloxy]-
4-octadecene 45a (0.675 g, 0.53 mmol) in dichloro-
methane (12 mL) and water (1.5 mL) was treated with
2,3-dichloro-5,6-dicyano-1,4-benzoquinone (180 mg,
0.796 mmol) at 5 ꢁC. The mixture was vigorously stirred
at 5 ꢁC for 30 min and then at 23 ꢁC for 1 h. The mixture
was diluted with dichloromethane (100 mL) and washed
with saturated sodium bicarbonate, 10% aqueous
sodium thiosulfate (1:1) (60 mL), 10% aqueous sodium
thiosulfate (60 mL) and brine (60 mL). The organic
phase was dried over anhydrous magnesium sulfate, fil-
tered and concentrated. The residue was purified by silica
gel chromatography (30–50% ethyl acetate/hexane) and
afforded the title compound (0.549 g, 90%) as a beige
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[3-
O-(5,5-di-tert-butyloxycarbonylpentyl)-2-O-benzoyl-a-D-
galactopyranosyloxy]-4-octadecene 51a. (2S,3R,4E)-3-
Benzoyloxy-2-hexadecanoylamino-1-[3-O-(5,5-di-tert-
butyloxycarbonylpentyl)-4,6-O-benzylidene-2-O-benzoyl-a-
d-galactopyranosyloxy]-4-octadecene
50a
(0.241 g,
0.19 mmol) was reacted by the procedure used to syn-
thesize compound 40a and afforded the title compound
1
(0.177 g, 80%) as a colorless gum. H NMR (400 MHz,
CDCl3) d 0.89 (6H, t, J=6.7 Hz, 2Â-CH3), 1.24–1.31
(46H, m, -(CH2)13- and -(CH2)10-), 1.38–1.47 (2H, m,
-CH2-), 1.44 (18H, s, 2Â-OtBu), 1.50–1.57 (4H, m,
-(CH2)2-), 1.66–1.81 (2H, m, -CH2-CH(CO2tBu)2),
1.98–2.14 (4H, m, ¼CH-CH2- and -NHCOCH2-), 2.48
(1H, br s, -OH-60), 2.78 (1H, s, -OH-40), 2.99 (1H, t,
J=7.5 Hz, -CH(CO2tBu)2), 3.56 (1H, dt, J=9.3, 6.2 Hz,
-OCH2-), 3.67 (1H, dt, J=9.3, 6.5 Hz, -OCH2-), 3.75
(1H, dd, J=11.1, 6.0 Hz, H-1), 3.81 (1H, dd, J=11.1,
3.9 Hz, H-1), 3.84 (1H, dd, J=10.3, 3.3 Hz, H-30), 3.85–
3.91 (1H, m, H-60), 3.96–4.01 (2H, m, H-60 and H-50),
4.20 (1H, br d, H-40), 4.54 (1H, m, H-2), 5.20 (1H, d,
J=3.8 Hz, H-10), 5.31 (1H, dd, J=10.1, 3.8 Hz, H-20),
5.49 (1H, dd, J=15.0, 7.3 Hz, H-4), 5.55 (1H, br t, H-3),
5.81 (1H, d, J=10.0 Hz, -NH-), 5.82 (1H, dt overlapped
by -NH-, H-5), 7.40–7.46, 7.54–7.60 and 7.98–8.01
(10H, 3 sets of m, aromatic H). IR (CH2Cl2) vmax
(cmÀ1) 3700–3430 (NH and OH), 3050, 2980, 2930 and
2850 (C–H), 1720, 1675 (C¼O).
1
amorphous solid. H NMR (400 MHz, CDCl3) d 0.89
(6H, t, J=6.7 Hz, 2Â-CH3), 1.26 (46H, m, -(CH2)13-
and -(CH2)10-), 1.40–1.52 (2H, m, -CH2-), 1.45 and 1.46
(18H, 2 s, 2Â-OtBu), 1.59–1.70 (4H, m, -(CH2)2-), 1.83
and 2.20 (2Â2H, 2 sets of m, -NHCOCH2- and -CH2-
CH(CO2tBu)2), 2.05 (2H, dt J=6.9, 6.9 Hz,=CH-CH2-),
2.50 (1H, br s, -OH), 3.11 (1H, t, J=7.5 Hz, -CH-
(CO2tBu)2), 3.54 (1H, dt, J=9.0, 6.6 Hz, -OCH2-), 3.62
(1H, dd, J=10.1 3.3 Hz, H-30), 3.69 (1H, dt, J=9.0,
6.6 Hz, -OCH2-), 3.74 (1H, br s, H-50), 3.76 (1H, dd,
J=11.0, 6.1 Hz, H-1), 3.93 (1H, dd, J=11.0, 3.8 Hz, H-
1), 4.06–4.13 (1H, m, H-20), 4.08 (1H, dd, J=12.4,
1.2 Hz, H-60), 4.26 (1H, dd, J=12.4, 1.0 Hz, H-60), 4.32
(1H, br d, H-40), 4.55 (1H, m, H-2), 4.99 (1H, d,
J=3.7 Hz, H-10), 5.53 (1H, dd, J=15.3, 7.4 Hz, H-4),
5.54 (1H, s, -OCHO-), 5.63 (1H, br t, H-3), 5.88 (1H, dt,
J=15.3, 6.9 Hz, H-5), 5.90 (1H, d, J=9.3 Hz, -NH-),
7.30–7.38, 7.45–7.58, 7.58–7.62 and 8.03–8.05 (10H, m,
aromatic H). IR (CH2Cl2) ꢀmax (cmÀ1) 3700, 3600 3440
(NH and OH), 3050, 2980, 2930 and 2860 (C-H), 1720,
1675 (C¼O). Anal. calcd for C69H111NO13: C, 71.28; H,
9.62; N, 1.20. Found: C, 71.21; H, 9.52; N, 1.34.
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[3-
O-(5,5-di-carboxypentyl)-2-O-benzoyl-a-D-galactopyr-
anosyloxy]-4-octadecene 52a. (2S,3R,4E)-3-Benzoyloxy-
2-hexadecanoylamino-1-[3-O-(5,5-di-tert-butyloxycarb-
onylpentyl)-2-O-benzoyl-a-d-galactopyranosyloxy]-4-
octadecene 51a (0.175 g, 0.148 mmol) was reacted by the
procedure used to synthesize compound 33a and affor-
ded the title compound (0.094 g, 57%) as a white solid.
1H NMR (400 MHz, pyridine-d5) d 0.85 (6H, br t,
2Â -CH3), 1.24 (46H, m, -(CH2)13- and -(CH2)10-),
1.56–1.63, 1.77–1.88, 1.99–2.04 and 2.20 (4H and 3Â2H,
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[3-
O-(5,5-di-tert-butyloxycarbonylpentyl)-4,6-O-benzyli-
dene-2-O-benzoyl-ꢁ-D-galactopyranosyloxy]-4-octadecene
50a. (2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-
[3-O-(5,5-di-tert-butyloxycarbonylpentyl)-4,6-O-benzyl-
idene-a-d-galactopyranosyloxy]-4-octadecene 48a (0.260 g,

1418
A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
.
Anal. calcd for C77H119NO14 0.3 H2O: C, 71.79; H,
9.36; N, 1.09. Found: C, 71.84; H, 9.26; N, 1.20.
-(CH2)2-, -CH2-, -CH2-CH(CO2H)2 and ¼CH-CH2-),
2.41 (2H, t, J=7.4 Hz, -NHCOCH2-), 3.57–3.50 (1H, m,
-OCH2-), 3.69–3.72 (2H, m, -OCH2- and -CH(CO2H)2),
4.04 (1H, dd, J=10.3, 6.9 Hz, H-1), 5.28 (1H, dd,
J=10.4, 1.9 Hz, H-30), 4.41–4.50 (3H, m, H-60 and H-1),
4.58 (1H, t, J=5.9 Hz, H-5), 4.71 (1H, br s, H-40), 5.23
(1H, m, H-2), 5.62 (1H, d, J=3.5 Hz, H-10), 5.92 (1H,
dd, J=15.4, 7.3 Hz, H-4), 6.07 (1H, dt, J=15.4, 6.5 Hz,
H-5), 6.13 (1H, dd, J=10.4, 3.4 Hz, H-20), 6.24 (1H, br
t, H-3), 7.36–7.40, 7.47–7.52, 8.16–8.18 and 8.36–8.71
(10H, 4 sets of m, aromatic H), 8.85 (1H, d, J=8.9 Hz,
-NH-). IR (nujol) ꢀmax (cmÀ1): 3650–3100 (broad, NH
and OH), 2930 and 2850 (C-H), 1720, 1650 (C¼O). MS
(ESIÀ) 1064.7 (M–H)À. MS (ESI+) 1088.6 (M+Na)+.
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[3-O-
(5,5-di-tert-butyloxycarbonylpentyl)-4,6-O-benzylidene-ꢀ-
D-galactopyranosyloxy]-4-octadecene 48b. (2S,3R,4E)-3-
Benzoyloxy-2-hexadecanoylamino-1-[3-O-(5,5-di-tert-
butyloxycarbonylpentyl)-2-O-p-methoxybenzyl-4,6-O-
benzylidene-b-d-galactopyranosyloxy]-4-octadecene 45b
(0.502 g, 0.39 mmol) was reacted by the procedure
used to synthesize compound 48a and afforded the
title compound (0.403 g, 89%) as an amorphous beige
solid. 1H NMR (400 MHz, CDCl3) d 0.89 (6H, t,
J=6.8 Hz, 2Â-CH3), 1.24–1.36 (46H, m, -(CH2)13-
and -(CH2)10-), 1.38–1.50 (2H, m, -CH2-), 1.45 and
1.45 (18H, 2 s, 2Â-OtBu), 1.53–1.71 (4H, m, -(CH2)2-),
1.77–1.85, 1.96–2.05 and 2.16 (3Â2H, -NHCOCH2-,
¼CH-CH2- and -CH2-CH(CO2tBu)2), 3.10 (1H, t,
J=7.4 Hz, -CH-(CO2tBu)2), 3.12 (1H, br s, -OH), 3.34
(1H, dd, J=9.6, 3.4 Hz, H-30), 3.42 (1H, br s, H-50),
3.54 (1H, dt, J=9.1, 6.6 Hz, -OCH2-), 3.71 (1H, dt,
J=9.1, 6.6 Hz, -OCH2-), 3.81 (1H, dd, J=11.4, 3.9 Hz,
H-1), 3.89 (1H, dd, J=9.6, 7.8 Hz, H-20), 4.07 (1H,
dd, J=12.3, 1.3 Hz, H-60), 4.18 (1H, dd, J=11.4,
4.8 Hz, H-1), 4.26 (1H, d, J=3.2 Hz, H-40), 4.30 (1H,
d, J=12.3 Hz, H-60), 4.34 (1H, d, J=7.8 Hz, H-10), 4.49
(1H, m, H-2), 5.51 (1H, dd, J=15.3, 7.2 Hz, H-4),
5.53 (1H, s, -OCHO-), 5.62 (1H, br t, H-3), 5.87 (1H, dt,
J=15.3, 6.7 Hz, H-5), 6.42 (1H, d, J=9.1 Hz,
-NH-), 7.30–7.37, 7.41–7.45, 7.48–7.57 and 8.03–8.05
(10H, m, aromatic H). IR (CH2Cl2) ꢀmax (cmÀ1) 3550–
3300 (NH and OH), 3050, 2980, 2930 and 2860 (C–H),
1720, 1670 (C¼O). Anal. calcd for C69H111NO13: C,
71.28; H, 9.62; N, 1.20. Found: C, 71.08; H, 9.54; N,
1.35.
1
Sodium salt: H NMR (400 MHz, CD3OD) d 0.84 (6H,
t, J=6.8 Hz, 2Â-CH3), 1.20–1.28 (48H, m, -(CH2)13-
and -(CH2)11-), 1.39–1.54, 1.79 and 1.93–2.07 (4H, 2H
and 4H, -(CH2)2-, -CH2-CH(CO2Na)2, ¼CH-CH2- and
-NHCOCH2-), 2.96 (1H, br t, -CH(CO2Na)2), 3.43–3.48
(1H, m, -OCH2-), 3.53 (1H, dd, J=10.6, 6.0 Hz, H-1),
3.63 (1H, m, -OCH2-), 3.67 and 3.71 (2H, 2 d,
J=11.4 Hz, H-60), 3.82 (1H, dd, J=10.5, 3.5 Hz, H-30),
3.85–3.89 (2H, m, H-50 and H-1), 4.12 (1H, br s, H-40),
4.74 (1H, m, H-2), 5.03 (1H, d, J=3.7 Hz, H-10), 5.19
(1H, dd, J=10.3, 3.7 Hz, H-20), 5.40–5.49 (2H, m, H-3
and H-4), 5.77 (1H, dt, J=14.3, 6.7 Hz, H-5), 7.34–
7.42, 7.50–7.55, 7.84–7.86 and 7.91–7.93 (10H, 4 sets
of m, aromatic H). IR (nujol) ꢀmax (cmÀ1) 3650-3100
(broad, NH and OH), 2920 and 2850 (C–H), 1720, 1595
(C¼O).
Synthesis of (2S,3R,4E)-3-benzoyloxy-2-hexadecanoyl-
amino-1-[3-O-(5,5-di-carboxypentyl)-2-O-benzoyl-ꢀ-D-
galactopyranosyloxy]-4-octadecene 52b
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[3-
O-(5,5-di-tert-butyloxycarbonylpentyl)-2-O-p-methoxy-
benzyl-4,6-O-benzylidene-ꢀ-D-galactopyranosyloxy]-4-
octadecene 45b. (2S,3R,4E)-3-Benzoyloxy-2-azido-1-[3-
O-(5,5-di-tert-butyloxycarbonylpentyl)-2-O-p-methoxy-
benzyl-4,6-O-benzylidene-b-d-galactopyranosyloxy]-4-
octadecene 44b was reacted by the procedure used to
synthesize compound 30a and afforded the title com-
pound (0.506 g, 68%) as a white solid. 1H NMR
(400 MHz, CDCl3) d 0.87–0.91 (6H, m, 2Â-CH3), 1.23–
1.31 (46H, m, -(CH2)13- and -(CH2)10-), 1.39–1.52 (4H, m,
-(CH2)2-), 1.44 and 1.45 (18H, 2 s, 2Â-OtBu), 1.64–1.71,
1.76–1.91 and 1.96–2.03 (2H, 4H and 2H, -NHCOCH2-,
¼CH-CH2-, -CH2-CH(CO2tBu)2 and -CH2-), 3.08 (1H,
t, J=7.5 Hz, -CH(CO2tBu)2), 3.36 (1H, br s, H-50), 3.43
(1H, dd, J=9.6, 3.5 Hz, H-30), 3.57 (1H, dt, J=9.1,
6.6 Hz, -OCH2-), 3.60–3.79 (2H, m, H-1 and -OCH2-),
3.76 (1H, dd, J=9.6, 7.8 Hz, H-20), 3.79 (3H, s, -OCH3),
4.05 (1H, dd, J=11.6, 1.2 Hz, H-60), 4.22–4.29 (1H, d
overlapped by H-1 and H-40, H-60), 4.24 (1H, dd,
J=11.1, 3.2 Hz, H-1), 4.25 (1H, d, J=3.5 Hz, H-40),
4.32 (1H, d, J=7.8 Hz, H-10), 4.45 (1H, m, H-2), 4.64
and 4.78 (2H, 2 d, J=10.4 Hz, -OCH2Ar), 5.48 (1H, dd,
J=15.3, 7.3 Hz, H-4), 5.54 (1H, s, -OCHO-), 5.60 (1H,
t, J=7.3 Hz, H-3), 5.83 (1H, dt, J=15.3, 6.7 Hz, H-5),
6.19 (1H, d, J=9.1 Hz, -NH-), 6.85–6.87, 7.22–7.28,
7.32–7.45, 7.52–7.56 and 8.05–8.07 (14H, 5 sets of m,
aromatic H). IR (CH2Cl2) ꢀmax (cmÀ1): 3440 (NH),
3050, 2990, 2930 and 2860 (C–H), 1720, 1670 (C¼O).
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[3-
O-(5,5-di-tert-butyloxycarbonylpentyl)-4,6-O-benzyli-
dene-2-O-benzoyl-ꢀ-D-galactopyranosyloxy]-4-octade-
cene 50b. (2S,3R,4E)-3-Benzoyloxy-2-hexadecanoyl-
amino-1-[3-O-(5,5-di-tert-butyloxycarbonylpentyl)-4,6-O-
benzylidene-b-d-galactopyranosyloxy]-4-octadecene 48b
(0.350 g, 0.305 mmol) was reacted by the procedure used
to synthesize compound 18 and afforded the title com-
pound (0.277 g, 72%) as a white gummy solid. 1H NMR
(400 MHz, CDCl3) d 0.89 (6H, br t, 2Â-CH3), 1.11–1.40
and 1.47–1.57 (52H, 2 sets of m, -(CH2)13-, -(CH2)11-
and -(CH2)2-), 1.41 and 1.42 (18H, 2 s, 2Â-OtBu),
1.65 (2H, m, -CH2-CH(CO2tBu)2), 1.79 (2H, br t,
-NHCOCH2-), 1.99 (2H, qa, J=6.9 Hz, ¼CH-CH2-),
2.87 (1H, t, J=7.5 Hz, -CH(CO2tBu)2), 3.45 (1H, dt,
J=9.5, 6.6 Hz, -OCH2-), 3.49 (1H, br s, H-50), 3.62–3.68
(1H, m overlapped by H-30, -OCH2-), 3.66 (1H, dd,
J=10.0, 3.2 Hz, H-30), 3.78 (1H, dd, J=10.4, 4.0 Hz, H-
1), 4.09 (1H, dd overlapped by H-1, H-60), 4.12 (1H, dd,
J=10.4, 4.3 Hz, H-1), 4.27 (1H, d, J=12.0 Hz, H-60),
4.33 (1H, d, J=3.2 Hz, H-40), 4.41 (1H, m, H-2), 4.61
(1H, d, J=7.9 Hz, H-10), 5.49 (1H, dd, J=15.3, 7.1 Hz,
H-4), 5.53 (1H, dd, J=10.0, 8.0 Hz, H-20), 5.57 (1H, s,
-OCHO-), 5.58 (1H, t, J=6.9 Hz, H-3), 5.81 (1H, dt,
overlapped by -NH-, H-5), 5.84 (1H, d, J=8.8 Hz, -NH-),
7.35–7.58 and 8.00–8.05 (15H, 2 sets of m, aromatic H).
IR (CH2Cl2) ꢀmax (cmÀ1) 3050, 2980, 2930, 2850 (C–H),

A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
1419
1722, 1670 (C¼O). Anal. calcd for C76H125NO14: C,
72.06; H, 9.15; N, 1.11. Found: C, 72.30; H, 9.12; N,
1.27.
and H-30), 3.93 (1H, dd, J=9.9, 6.3 Hz, H-60), 4.06 (1H,
br s, H-40), 4.33 (1H, m, H-2), 4.56 (1H, d, J=8.0 Hz,
H-10), 5.29 (1H, br dd, H-20), 5.38 (1H, dd, J=15.0,
7.7 Hz, H-4), 5.44 (1H, br t, H-3), 5.66 (1H, dt, J=15.0,
6.9 Hz, H-5), 7.37–7.43, 7.51–7.53 and 7.91–7.98 (10H, 3
sets of m, aromatic H). IR (nujol) ꢀmax (cmÀ1) 3700–
2400 (O–H, N–H), 2930, 2850 (broad, C–H), 1720,
1650, 1600 (C¼O).
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[3-
O-(5,5-di-tert-butyloxycarbonylpentyl)-2-O-benzoyl-b-D-
galactopyranosyloxy]-4-octadecene 51b. (2S,3R,4E)-3-
Benzoyloxy-2-hexadecanoylamino-1-[3-O-(5,5-di-tert-
butyloxycarbonylpentyl)-4,6-O-benzylidene-2-O-benzoyl-b-
d-galactopyranosyloxy]-4-octadecene
50b
(0.257 g,
Synthesis of (2S,3R,4E)-3-benzoyloxy-2-hexadecanoyla-
mino-1-[3-O-(5,5-di-carboxypentyl)-ꢁ-D-galactopyrano-
syloxy]-4-octadecene 54a
0.2 mmol) was reacted by the procedure used to synthe-
size compound 40a and afforded the title compound
1
(0.190 g, 81%) as a white solid. H NMR (400 MHz,
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[3-
O-(5,5-di-tert-butyloxycarbonylpentyl)-ꢁ-D-galactopyr-
anosyloxy]-4-octadecene 53a. (2S,3R,4E)-3-Benzoyloxy-
2-hexadecanoylamino-1-[3-O-(5,5-di-tert-butyloxycarb-
onylpentyl)-4,6-O-benzylidene-a-d-galactopyranosyl-
oxy]-4-octadecene 48a (0.260 g, 0.22 mmol) was reacted
by the procedure used to synthesize compound 40a and
afforded the title compound (0.173 g, 74%) as a gummy
CDCl3) d 0.89 (6H, br t, 2Â-CH3), 1.24–1.75 (55H, m,
-(CH2)13-, -(CH2)11-, -(CH2)2-, -CH2-CH(CO2tBu)2 and
-OH), 1.44 and 1.43 (18H, 2 s , 2Â-OtBu), 1.92 (2H,
J=7.4 Hz, -NHCOCH2-), 2.01 (2H, dt, J=6.9, 6.9 Hz,
¼CH-CH2-), 2.93 (1H, t, J=7.4 Hz, -CH-(CO2tBu)2),
3.09 (1H, br s, -OH), 3.46 (1H, dt, J=9.5, 6.4 Hz,
-OCH2-), 3.43–3.53 and 3.62–3.67 (2Â2H, 2 sets of m,
-OCH2-, H-1 and H-50), 3.73 (1H, dd, J=12.2, 3.6 Hz,
H-60), 3.93 (1H, dd, J=12.2, 6.2 Hz, H-60), 3.98 (1H, dd,
J=9.8, 2.6 Hz, H-30), 4.12 (1H, br d, H-40), 4.44 (1H, m,
H-2), 4.48 (1H, d, J=7.9 Hz, H-10), 5.42 (1H, dd,
J=9.7, 8.1 Hz, H-20), 5.49 (1H, dd, J=15.3, 7.9 Hz, H-
4), 5.67 (1H, t, J=7.8 Hz, H-3), 5.82 (1H, d, J=9.4 Hz,
-NH-), 5.89 (1H, dt, J=15.3, 6.9 Hz, H-5), 7.33–7.47,
7.52–7.60 and 8.02–8.06 (10H, 3 sets of m, aromatic H).
IR (CH2Cl2) ꢀmax (cmÀ1) 3700–3400 (O–H), 3050, 2980,
2930, 2850 (C–H), 1722, 1670 (C¼O).
1
white solid. H NMR (400 MHz, CDCl3) d 0.89 (6H, t,
J=6.7 Hz, 2Â -CH3), 1.26–1.37 (46H, m, -(CH2)13- and
-(CH2)10-), 1.41–1.51 (2H, m, -CH2-), 1.47 (18H, s,
2Â-OtBu), 1.53–1.73 (4H, m, -(CH2)2-), 1.82–1.88 (2H,
m, -CH2-CH(CO2tBu)2), 2.05 (2H, dt, J=6.9, 6.9 Hz,
¼CH-CH2-), 2.20 (2H, m, -NHCOCH2-), 2.68 (1H, br s,
-OH), 3.15 (1H, t, J=7.4 Hz, -CH(CO2tBu)2), 3.45 (1H,
dd, J=9.7, 3.2 Hz, H-30), 3.62–3.71 (2H, m, -OCH2-),
3.77 (1H, dd, J=11.1, 5.7 Hz, H-1), 3.82–3.99 (5H, m,
H-1, H-20, H-50 and H-60), 4.12 (1H, d, J=3.0 Hz, H-40),
4.56 (1H, m, H-2), 4.90 (1H, d, J=3.9 Hz, H-10), 5.53
(1H, dd, J=15.3, 7.4 Hz, H-4), 5.63 (1H, br t, H-3), 5.90
(1H, dt, J=15.3, 6.9 Hz, H-5), 5.96 (1H, d, J=9.3 Hz,
-NH-), 7.45–7.49, 7.58–7.62 and 8.03–8.05 (5H, 3 sets of
m, aromatic H). IR (CH2Cl2) ꢀmax (cmÀ1) 3700–3400 (NH
and OH), 3050, 2980, 2930 and 2860 (C–H), 1720, 1675
(C¼O).
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[3-
O-(5,5-di-carboxypentyl)-2-O-benzoyl-ꢀ-D-galactopyr-
anosyloxy]-4-octadecene 52b. (2S,3R,4E)-3-Benzoyloxy-
2-hexadecanoylamino-1-[3-O-(5,5-di-tert-butyloxycar-
bonylpentyl)-2-O-benzoyl-b-d-galactopyranosyloxy]-4-
octadecene 51b (0.182 g, 0.15 mmol) was reacted by the
procedure used to synthesize compound 33a and affor-
ded the title compound (0.155 g, 97%) as a beige solid.
1H NMR (400 MHz, pyridine-d5) d 0.85 (6H, t,
J=6.7 Hz, 2Â-CH3), 1.20–1.29 and 1.49–1.75 (46H and
6H, 2 sets of m, -(CH2)13-, -(CH2)11- and -(CH2)2-), 1.93
and 2.07–2.21 (2H and 4H, 2 sets of m, -NHCOCH2-,
¼CH-CH2-, -CH2-CH(CO2H)2), 3.46 (1H, m, -OCH2-),
3.62 (1H, t, J=7.4 Hz, -CH(CO2H)2), 3.71 (1H,
m, -OCH2-), 3.86 (1H, dd, J=9.9, 2.9 Hz, H-30), 4.11
(1H, dd, J=6.2, 6.2 Hz, H-50), 4.30 (1H, dd, J=10.6,
5.1 Hz, H-1), 4.37–4.42 (2H, m, H-1 and H-60), 4.48
(1H, dd, J=10.8, 6.2 Hz, H-60), 4.68 (1H, d, J=2.7 Hz,
H-40), 5.11 (1H, m, H-2), 5.13 (1H, d, J=7.9 Hz, H-10),
5.80 (1H, dd, J=15.5, 7.2 Hz, H-4), 5.92 (1H, dt,
J=15.5, 6.6 Hz, H-5), 6.13 (1H, br t, H-3), 6.26 (1H, dd,
J=9.8, 8.1 Hz, H-20), 7.36–7.39, 7.45–7.54, 8.19–8.21
and 8.32–8.36 (11H, 4 sets of m, aromatic H and -NH-).
IR (nujol) ꢀmax (cmÀ1) 3700–2400 (O–H, N–H), 2930,
2850 (broad, C-H), 1720, 1650 (C¼O). MS (ESIÀ)
1064.4 (M–H)À. MS (ESI+) 1088.7 (M+Na)+. Sodium
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[3-
O-(5,5-di-carboxypentyl)-a-D-galactopyranosyloxy]-4-
octadecene 54a. (2S,3R,4E)-3-Benzoyloxy-2-hexade-
canoylamino-1-[3-O-(5,5-di-tert-butyloxycarbonylpentyl)-
a-d-galactopyranosyloxy]-4-octadecene 53a (0.165 g,
0.154 mmol) was reacted by the procedure used to syn-
thesize compound 33a and afforded the title compound
1
(0.124 g, 80%) as a white solid. H NMR (400 MHz,
pyridine-d5) d 0.85 (6H, t, J=6.7 Hz, 2Â -CH3), 1.24–
1.36 (46H, m, -(CH2)13- and -(CH2)10-), 1.64–1.75 (4H,
m, -(CH2)2-), 1.84 (2H, m, -CH2-), 2.04 (2H, dt, J=6.8,
6.8 Hz,=CH-CH2-), 2.28 (2H, m, -CH2-CH(CO2H)2),
2.47 (2H, t, J=7.4 Hz, -NHCOCH2-), 3.51–3.53 and
3.72–3.74 (2Â 1H, 2 sets of m, -OCH2-), 3.83 (1H, t,
J=7.4 Hz, -CH(CO2H)2), 3.96 (1H, dd, J=10.0, 3.0 Hz,
H-3-), 4.17 (1H, dd, J=10.8, 6.6 Hz, H-60), 4.35–4.53
(4H, m, H-60, H-1 and H-50), 4.60 (1H, br d, H-40), 4.68
(1H, dd, J=10.0, 3.8 Hz, H-20), 5.22 (1H, m, H-2), 5.36
(1H, d, J=3.8 Hz, H-10), 5.97 (1H, dd, J=15.5, 7.1 Hz,
H-4), 6.13 (1H, dt, J=15.5, 6.8 Hz, H-5), 6.36 (1H, br t,
H-3), 7.39–7.43, 7.50–7.53 and 8.24–8.26 (5H, 3 sets
of m, aromatic H), 8.91 (1H, d, J=8.7 Hz, -NH-). IR
(nujol) ꢀmax (cmÀ1) 3650–3100 (NH and OH), 2930, 2850
(C-H), 1730, 1640 (C¼O). MS (ESIÀ) 960.6 (M–H)À.
MS (ESI+) 962.7 (MH+), 984.7 (M+Na)+. Sodium
1
salt: H NMR (400 MHz, CD3OD) d 0.83 (6H, br t,
2Â-CH3), 1.14–1.46 (52H, m, -(CH2)13-, -(CH2)11-
and -(CH2)2-), 1.67 and 1.80–1.95 (2H and 4H, 2 sets
of m, -NHCOCH2-, -CH2-CH(CO2Na)2 and ¼CH–
CH2-), 2.93 (1H, t, J=7.1 Hz, -CH(CO2Na)2), 3.38 (1H,
m, -OCH2-), 3.54–3.72 (6H, m, -OCH2-, H-60, H-1, H-50

1420
A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
salt: IR (nujol) ꢀmax (cmÀ1) 3650-3100 (broad, NH and
OH), 2920 and 2850 (broad, C-H), 1710, 1640 and 1585
(C¼O).
ded the title compound (0.308 g, 84%) as a white amor-
1
phous solid. H NMR (400 MHz, CDCl3) d 0.89 (6H, t,
J=6.7 Hz, 2Â-CH3), 1.25–1.35 (46H, m, -(CH2)13- and
-(CH2)10-), 1.40–1.49 (2H, m, -CH2-), 1.47 (18H, s,
2Â-OtBu), 1.53–1.71 (4H, 2 sets of m, -(CH2)2-), 1.84,
2.04 and 2.19 (3Â2H, 3 sets of m, -NHCOCH2-, ¼CH–
CH2- and -CH2-CH(CO2tBu)2), 2.77 and 2.90 (2H, 2 br
s, 2Â -OH), 3.13–3.17 (1H, br s, -OH), 3.15 (1H, t,
J=7.4 Hz, -CH(CO2tBu)2), 3.24 (1H, dd, J=9.4 and
3.2 Hz, H-30), 3.45 (1H, br dd, H-50), 3.59 (1H, dt,
J=9.2, 6.5 Hz, -OCH2-), 3.68–3.77 (3H, m, -OCH2-,
H-60 and H-20), 3.82 (1H, dd, J=11.2, 4.3 Hz, H-1), 3.93
(1H, dd, J=12.0, 6.0 Hz, H-60), 4.02–4.06 (2H, m, H-1
and H-40), 4.30 (1H, d, J=7.7 Hz, H-10), 4.56 (1H, m,
H-2), 5.51 (1H, dd, J=15.3, 7.5 Hz, H-4), 5.65 (1H, br t,
H-3), 5.90 (1H, dt, J=15.3, 6.7 Hz, H-5), 6.13 (1H, d,
J=9.4 Hz, -NH-), 7.44–7.48, 7.53–7.61 and 8.03–
8.05 (5H, 3 sets of m, aromatic H). IR (CH2Cl2) ꢀmax
(cmÀ1) 3700–3300 (NH and OH), 3050, 2980, 2930 and
2850 (C–H), 1720, 1670 (C¼O). Anal. calcd for
Synthesis of (2S,3R,4E)-3-hydroxy-2-hexadecanoyl-
amino-1-[3-O-(5,5-di-carboxypentyl)-ꢁ-D-galactopyrano-
syloxy]-4-octadecene 57a. A solution of (2S,3R,4E)-3-
benzoyloxy-2-hexadecanoylamino-1-[3-O-(5,5-di-carb-
oxypentyl)-a-d-galactopyranosyloxy]-4-octadecene 54a
(0.291 g, 0.3 mmol) in dry dichloromethane (20 mL) was
treated with a solution of sodium methoxide in metha-
nol (0.0276 g of sodium in 1.5 mL of methanol) at 5 ꢁC.
The mixture was stirred at 23 ꢁC for 1 h, then cooled
down to 5 ꢁC and treated with Dowex 50W8 resin H+
until the pH reached ꢀ6.0–6.5. The resin was filtered
and washed with dichloromethane/methanol (1:1,
3Â5 mL). The combined filtrate and washings were
concentrated and co-evaporated with toluene (4Â5 mL).
The residue was purified by silica gel chromatography
(30 g, 0–10% methanol/ethyl acetate and traces of pyri-
dine then 11:1:88 to 50:20:30 methanol/water/ethyl ace-
tate) to give the title compound (0.152 g, 56%) as a
white solid. 1H NMR (400 MHz, pyridine-d5) d 0.85
(6H, m, 2Â-CH3), 1.25–1.38, 1.68, 1.85 and 2.08
(46H, 4H, 2H and 2H, 4 sets of m, -(CH2)13-, -(CH2)11-,
-(CH2)2- and -CH2CH(CO2H)2), 2.28 (2H, dt, J=7.2,
7.2 Hz, ¼CH-CH2-), 2.48 (2H, m, -NHCOCH2-), 3.50
and 3.72 (2Â1H, 2 sets of m, -OCH2-), 3.82 (1H, t,
J=7.4 Hz, -CH(CO2H)2), 3.93 (1H, dd, J=9.9, 3.0 Hz,
H-30), 4.35 (1H, dd, J=10.5, 5.2 Hz, H-1 or H-60), 4.41–
4.51 (4H, m, H-1, H-50 and H-60), 4.61 (1H, br d,
J=2.6 Hz, H-40), 4.66 (1H, dd, J=9.9, 3.9 Hz, H-20),
4.83–4.89 (2H, m, H-2 and H-3), 5.41 (1H, d, J=3.9 Hz,
H-10), 6.00 and 6.03 (2Â1H, 2 d, J=15.4 Hz, H-4 and
H-5), 8.59 (1H, d, J=8.3 Hz, -NH-). IR (nujol) ꢀmax
(cmÀ1) 3650–3100 (NH and OH), 2920, 2850 (C–H),
.
C62H107NO13 0.5H2O: C, 68.73; H, 10.05; N, 1.29.
Found: C, 68.79; H, 9.92; N, 1.45.
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[3-
O-(5,5-di-carboxypentyl)-ꢀ-D-galactopyranosyloxy]-4-
octadecene 54b. (2S,3R,4E)-3-Benzoyloxy-2-hexadeca-
noylamino-1-[3-O-(5,5-di-tert-butyloxycarbonylpentyl)-b-
d-galactopyranosyloxy]-4-octadecene
53b
(0.290 g,
0.27 mmol) was reacted by the procedure used to syn-
thesize compound 33a and afforded the title compound
1
(0.142 g, 55%) as a white solid. H NMR (400 MHz,
pyridine-d5) d 0.86 (6H, t, J=6.6 Hz, 2Â-CH3), 1.24–
1.37 (46H, m, -(CH2)13- and -(CH2)10-), 1.70–1.86, 1.98
and 2.30 (6H and 2Â2H, -(CH2)2-, -CH2-, ¼CH-CH2- and
-CH2CH(CO2H)2), 2.44 (2H, t, J=7.4 Hz, -NHCOCH2-),
3.65 (1H, dd, J=9.3, 3.0 Hz, H-30), 3.63–3.71 (1H, m,
-OCH2-), 3.83 (1H, t, J=7.4 Hz, -CH(CO2H)2), 3.81–
3.87 (1H, m, -OCH2-), 4.02 (1H, dd, J=6.0, 6.0 Hz, H-
50), 4.15 (1H, dd, J=10.9, 4.4 Hz, H-1), 4.39 (1H, dd,
J=10.9, 6.0 Hz, H-60), 4.45 (1H, dd, J=10.9, 6.0 Hz, H-
60), 4.51 (1H, br dd, H-20), 4.57 (1H, dd, J=10.9, 5.1 Hz,
H-1), 4.59 (1H, br d, J=2.8 Hz, H-40), 4.89 (1H, d,
J=7.7 Hz, H-10), 5.11 (1H, m, H-2), 5.88 (1H, dd,
J=15.4, 7.4 Hz, H-4), 6.02 (1H, dt, J=15.4, 6.5 Hz, H-
5), 6.20 (1H, br t, H-3), 7.35–8.22 (5H, m, aromatic H),
1
1715, 1645 (C¼O). Sodium salt: H NMR (400 MHz,
CD3OD) d 0.85 (6H, t, J=6.8 Hz, 2Â-CH3), 1.24–1.42,
1.54–1.64, 1.78–1.84 (48H, 4H and 2H, 3 sets of m,
-(CH2)11- and -(CH2)13-, -(CH2)2- and -CH2CH-
(CO2Na)2), 1.96 (2H, dt J=6.8, 6.8 Hz, ¼CH-CH2-),
2.15 (2H, t, J=7.5 Hz, -NHCOCH2-), 3.06 (1H, t,
J=7.4 Hz, -CH(CO2Na)2), 3.46–3.53 and 3.64–3.73 (2H
and 5H, 2 sets of m, -OCH2-, H-1, H-30, H-50 and H-60),
3.77 (1H, dd, J=10.8, 3.8 Hz, H-1 or H-60), 3.82 (1H,
dd, J=10.0, 3.8 Hz, H-20), 3.87 (1H, dt, J=8.5, 3.4 Hz,
H-2), 4.04 (1H, br d, J=2.0 Hz, H-40), 4.08 (1H, t,
J=8.2 Hz, H-3), 4.81 (1H, d, J=3.9 Hz, H-10), 5.39 (1H,
dd, J=15.3, 7.5 Hz, H-4), 5.67 (1H, dt, J=15.3, 6.8 Hz,
H-5). IR (nujol) ꢀmax (cmÀ1) 3700–3050 (NH and OH),
2920, 2850 (C–H), 1645 (C¼O). MS (ESIÀ) 856.6 (M–
H)À. MS (ESI+) 858.7 (MH+), 880.6 (M+Na)+.
8.70 (1H, d, J=10.9 Hz, -NH-). IR (nujol) ꢀmax (cmÀ1
)
3700–3000 (NH and OH), 2920 and 2850 (broad, C-H),
1715, 1630 (C¼O). MS (ESIÀ) 960.6 (M–H)À. MS
(ESI+) 984.7 (M+Na)+. Sodium salt: 1H NMR
(400 MHz, CD3OD) d 0.89 (6H, t, J=6.8 Hz, 2Â-
CH3), 1.27–1.43 (48H, m, -(CH2)13- and -(CH2)11-),
1.58–1.66 (4H, m, -(CH2)2-), 1.84, 2.05, and 2.18–2.24
(3Â2H,
-NHCOCH2-,=CH-CH2-
and
-CH2-
Synthesis of (2S,3R,4E)-3-benzoyloxy-2-hexadecanoyl-
amino-1-[3-O-(5,5-di-carboxypentyl)-ꢀ-D-galactopyrano-
syloxy]-4-octadecene 54b
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-[3-
O-(5,5-di-tert-butyloxycarbonylpentyl)-ꢀ-D-galactopyr-
anosyloxy]-4-octadecene 53b. (2S,3R,4E)-3-Benzoyloxy-
2-hexadecanoylamino-1-[3-O-(5,5-di-tert-butyloxycarb-
onylpentyl)-4,6-O-benzylidene-b-d-galactopyranosyloxy]-4-
octadecene 48b (0.397 g, 0.34 mmol) was reacted by the
procedure used to synthesize compound 40a and affor-
CH(CO2Na)2), 3.09 (1H, t, J=7.1 Hz, -CH(CO2Na)2),
3.20 (1H, dd, J=9.6, 3.0 Hz, H-30), 3.42–3.75 (7H, m,
H-20, H-50, H-60, -OCH2- and H-1), 4.01 (1H, br d, H-
40), 4.13 (1H, dd, J=10.4, 5.0 Hz, H-1), 4.22 (1H, d,
J=7.7 Hz, H-10), 4.42 (1H, m, H-2), 5.51 (1H, dd,
J=15.0, 7.8 Hz, H-4), 5.58 (1H, br t, H-3), 5.89 (1H, dt,
J=15.0, 6.8 Hz, H-5), 7.45–7.48, 7.57–7.61 and 8.00–
8.02 (5H, 3 sets of m, aromatic H). IR (nujol) ꢀmax
(cmÀ1) 3700–3000 (NH), 2920 and 2850 (broad, C–H),
1715, 1585 (C¼O).

A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
1421
Synthesis of (2S,3R,4E)-3-benzoyloxy-2-hexadecanoyla-
mino-1-{3-O-[4-(2,2-di-carboxyethyl)benzyl]-ꢁ-D-galacto-
pyranosyloxy}-4-octadecene 56a
m, -NHCOCH2- and=CH-CH2-), 3.13 (2H, d,
J=7.8 Hz, -CH2-CH(CO2tBu)2), 3.44 (1H, t, J=7.8 Hz,
-CH(CO2tBu)2), 3.65 (1H, br s, H-50), 3.78 (3H, s,
-OCH3), 3.78–3.79 (2H, m, H-1), 3.92 (1H, dd, J=10.1,
3.4 Hz, H-30), 4.00 (1H, br d, H-60), 4.05 (1H, dd,
J=10.1, 3.5 Hz, H-20), 4.18–4.21 (2H, m, H-40 and H-
60), 4.48 (1H, m, H-2), 4.63–4.75 (4H, m, 2Â-OCH2Ar),
3.91 (1H, d, J=3.5 Hz, H-10), 5.47 (1H, s, -OCHO-),
5.48 (1H, dd, J=15.3, 7.5 Hz, H-4), 5.60 (1H, br t,
J=7.3 Hz, H-3), 5.79 (1H, dt, J=15.3, 6.9 Hz, H-5),
5.99 (1H, d, J=9.1 Hz, -NH-), 6.79–6.82, 7.17–7.21,
7.27–7.52, 7.57–7.60 and 8.04–8.06 (18H, 5 sets of m,
aromatic H).
(2S,3R,4E)-3-Benzoyloxy-2-azido-1-{3-O-[4-(2,2-di-
tert-butyloxycarbonylethyl)benzyl]-2-O-p-methoxy-
benzyl-4,6-O-benzylidene-ꢁ-D-galactopyranosyloxy}-4-
octadecene 46a and (2S,3R,4E)-3-benzoyloxy-2-azido-1-
{3-O-[4-(2,2-di-tert-butyloxycarbonylethyl)benzyl]-2-O-
p-methoxybenzyl-4,6-O-benzylidene-ꢀ-D-galactopyr-
anosyloxy}-4-octadecene 46b. Ethyl 2-O-p-methoxy-
benzyl-4,6-di-O-benzylidene-3-O-[4-(2,2-di-tert-butyloxy-
carbonylethyl)benzyl]-1-thio-b-d-galactopyranoside 27
(0.500 g, 0.666 mmol) and (2S,3R,4E)-2-azido-3-ben-
zoyloxy-4-octadecen-1-ol 28 (0.258 g, 0.6 mmol) were
reacted by the procedure used to synthesize compounds
29a and b except that acetonitrile was used as solvent.
This afforded the a-anomer (0.114 g, 15%) and the
b-anomer (0.111 g, 15%) of the title compound as oils.
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-{3-
O-[4-(2,2-di-tert-butyloxycarbonylethyl)benzyl]-4,6-O-
benzylidene-ꢁ-D-galactopyranosyloxy}-4-octadecene 49a.
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-{3-O-
[4-(2,2-di-tert-butyloxycarbonylethyl)benzyl]-2-O-p-meth-
oxybenzyl-4,6-O-benzylidene-a-d-galactopyranosyloxy}-
4-octadecene 47a (0.135 g, 0.103 mmol) was reacted by
the procedure used to synthesize compound 48a and
afforded the title compound (0.112 g, 91%) as a sticky
solid. 1H NMR (400 MHz, CDCl3) d 0.89 (6H, t,
J=6.7 Hz, 2Â -CH3), 1.26–1.42 (46H, m, -(CH2)11- and
-CH2)12-), 1.42 (18H, s, 2Â-OtBu), 1.60 (2H, m, -CH2-),
2.05 (2H, m, ¼CH-CH2-), 2.18 (2H, m, -NHCOCH2-),
2.36 (1H, br s, -OH), 3.12 (2H, d, J=7.9 Hz, -CH2–
CH(CO2tBu)2), 3.42 (1H, t, J=7.9 Hz, -CH(CO2tBu)2),
3.69 (1H, s, H-50), 3.71–3.76, 4.18–4.24 and 3.92 (2H,
2H, 1H, 2 sets of m and 1 dd, J=10.9, 3.7 Hz, H-20, H-
30, H-1 and H-40), 4.02 (1H, br d, J=12.1 Hz, H-60),
4.23 (1H, br d, H-60), 4.55 (1H, m, H-2), 4.70 and 4.72
(2Â1H, 2 d, J=12.3 Hz, -OCH2Ar), 4.99 (1H, d,
J=3.7 Hz, H-10), 5.45 (1H, s, -OCHO-), 5.52 (1H, dd,
J=15.3, 7.3 Hz, H-4), 5.62 (1H, br t, H-3), 5.85 (1H, d,
J=8.4 Hz, -NH-), 5.84–5.91 (1H, dt overlapped by
-NH-, H-5), 7.16–7.18, 7.31–7.40, 7.44–7.52, 7.58–7.61
and 8.03–8.05 (14H, 5 sets of m, aromatic H).
1
ꢁ-Anomer: H NMR (400 MHz, CDCl3) d 0.89 (3H, t,
J=6.7 Hz, -CH3), 1.26–1.36 (22H, m, -(CH2)11-), 1.42
(18H, s, 2Â-OtBu), 2.07 (2H, dt, J=6.9, 6.9 Hz, ¼CH–
CH2-), 3.13 (2H, d, J=7.8 Hz, -CH2-CH(CO2tBu)2),
3.45 (1H, t, J=7.8 Hz, -CH(CO2tBu)2), 3.59 (1H, dd,
J=10.9, 7.4 Hz, H-1), 3.65 (1H, br s, H-50), 3.73 (1H,
dd, J=10.9, 4.6 Hz, H-1), 3.79 (3H, s, -OCH3), 3.93–
3.98 (2H, m, H-2 and H-30), 4.02 (1H, d, J=12.2 Hz, H-
60), 4.06 (1H, dd, J=10.2, 3.4 Hz, H-20), 4.15 (1H, d,
J=3.0 Hz, H-40), 4.20 (1H, d, J=12.2 Hz, H-60), 4.60
and 4.76 (2Â1H, 2 d, J=11.6 Hz, -OCH2Ar), 4.69 and
4.79 (2Â1H, 2 d, J=12.1 Hz, -OCH2Ar), 4.89 (1H, d,
J=3.3 Hz, H-10), 5.48 (1H, s, -OCHO-), 5.58 (1H, dd,
J=14.7, 8.0 Hz, H-4), 5.63 (1H, dd, J=8.0, 4.1 Hz, H-
3), 5.91 (1H, dt, J=14.7, 6.9 Hz, H-5), 6.82–6.84, 7.17–
7.19, 7.27–7.39, 7.45–7.61 and 8.07–8.09 (18H, 5 sets of
m, aromatic H). ꢀ-Anomer: 1H NMR (400 MHz,
CDCl3) d 0.89 (3H, t, J=6.8 Hz, -CH3), 1.24–1.36
(22H, m, -(CH2)11-), 1.42 (2Â9H, 2 s, 2Â-OtBu), 2.04
(2H, dt, J=6.9, 6.9 Hz, ¼CH-CH2-), 3.13 (2H, d,
J=7.8 Hz, -CH2-CH(CO2tBu)2), 3.30 (1H, br s, H-50),
3.45 (1H, t, J=7.8 Hz, -CH(CO2tBu)2), 3.52 (1H, dd,
J=9.6, 3.5 Hz, H-30), 3.61 (1H, dd, J=9.7, 5.1 Hz, H-
1), 3.79 (3H, s, -OCH3), 3.85 (1H, dd, J=9.6, 7.7 Hz,
H-20), 3.98–4.07 (4H, m, H-1, H-2, H-40 and H-60), 4.28
(1H, d, J=12.0 Hz, H-60), 4.38 (1H, d, J=7.7 Hz,
H-10), 4.68–4.87 (4H, m, 2Â-OCH2Ar), 5.49 (1H, s,
-OCHO-), 5.58 (1H, dd, J=15.3, 8.0 Hz, H-4), 5.70
(1H, dd, J=8.0, 3.6 Hz, H-3), 5.90 (1H, dt, J=15.3,
6.9 Hz, H-5), 6.85–6.88, 7.16–7.18, 7.28–7.48, 7.55–
7.60 and 8.07–8.09 (18H, 5 sets of m, aromatic H).
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-{3-
O-[4-(2,2-di-tert-butyloxycarbonylethyl)benzyl]-a-D-
galactopyranosyloxy}-4-octadecene 55a. (2S,3R,4E)-3-
Benzoyloxy-2-hexadecanoylamino-1-{3-O-[4-(2,2-di-tert-
butyloxycarbonylethyl)benzyl]-4,6-O-benzylidene-a-d-
galactopyranosyloxy}-4-octadecene 49a (0.112 g, 0.09
mmol) was reacted by the procedure used to synthesize
compound 40a and afforded the title compound
1
(0.071 g, 68%) as an oil. H NMR (400 MHz, CDCl3)
d 0.89 (6H, t, J=6.7 Hz, 2Â-CH3), 1.26–1.37 (46H, m,
-(CH2)11- and -(CH2)12-), 1.43 (18H, s, 2Â-OtBu), 1.60
(2H, m, -CH2-), 2.05 (2H, dt, J=7.0, 7.0 Hz, ¼CH-CH2-),
2.20 (2H, m, -NHCOCH2-), 2.38 and 2.50 (2H and 1H,
2 br s, 2Â -OH), 3.14 (2H, d, J=7.8 Hz, -CH2–
CH(CO2tBu)2), 3.45 (1H, t, J=7.8 Hz, -CH(CO2tBu)2),
3.60 (1H, dd, J=9.7, 3.2 Hz, H-30), 3.73 (1H, dd,
J=11.1, 5.8 Hz, H-1), 3.80–3.93 (4H, m, H-1, H-60 and
H-50), 3.96 (1H, dd, J=9.6, 3.8 Hz, H-20), 4.03 (1H, br
d, J=2.9 Hz, H-40), 4.56 (1H, m, H-2), 4.66 and 4.79
(2Â1H, 2d, J=11.6 Hz, -OCH2Ar), 4.89 (1H, d,
J=3.9 Hz, H-10), 5.53 (1H, dd, J=15.3, 7.3 Hz, H-4),
5.63 (1H, br t, H-3), 5.89 (1H, dt, J=15.3, 7.0 Hz, H-5),
5.94 (1H, d, J=9.2 Hz, -NH-), 7.21–7.23, 7.29–7.31,
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-{3-
O-[4-(2,2-di-tert-butyloxycarbonylethyl)benzyl]-2-O-p-
methoxybenzyl-4,6-O-benzylidene-a-D-galactopyr-
anosyloxy}-4-octadecene 47a. (2S,3R,4E)-3-Benzoyloxy-
2-azido-1-{3-O-[4-(2,2-di-tert-butyloxycarbonylethyl)-
benzyl]-2-O-p-methoxybenzyl-4,6-O-benzylidene-a-d-
galactopyranosyloxy}-4-octadecene 46a (0.114g, 0.102
mmol) was reacted by the procedure used to synthesize
compound 30a and afforded the title compound
(0.099 g, 74%) as a white solid. 1H NMR (400
MHz, CDCl3) d 0.89 (6H, t, J=6.7 Hz, 2Â-CH3), 1.24–
1.26 (46H, m, -(CH2)11- and -(CH2)12-), 1.42 (18H, s,
2Â-OtBu), 1.48–1.61 (2H, m, -CH2-), 1.98–2.14 (4H,

1422
A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
7.45–7.49, 7.58–7.62 and 8.03–8.05 (9H, 5 sets of m,
aromatic H).
J=11.6 Hz, H-60), 4.31 (1H, d, J=7.8 Hz, H-10), 4.46
(1H, m, H-2), 4.68 and 4.80 (2Â1H, 2 d, 2Â-OCH2Ar),
4.71 and 4.74 (2Â1H, 2 d, J=12.2 Hz, -OCH2Ar), 5.48
(1H, s, -OCHO-), 5.48 (1H, dd, J=15.3, 7.4 Hz, H-4),
5.61 (1H, br t, H-3), 5.83 (1H, dt, J=15.3, 6.7 Hz, H-5),
6.16 (1H, d, J=9.1 Hz, -NH-), 6.84–6.86, 7.16–7.18,
7.25–7.31, 7.33–7.44, 7.52–7.56 and 8.05–8.07 (18H, 6
sets of m, aromatic H).
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-{3-
O-[4-(2,2-di-tert-butyloxycarbonylethyl)benzyl]-ꢁ-D-
galactopyranosyloxy}-4-octadecene 56a. (2S,3R,4E)-3-
Benzoyloxy-2-hexadecanoylamino-1-{3-O-[4-(2,2-di-tert-
butyloxycarbonylethyl)benzyl] - a - d - galactopyranosyl-
oxy}-4-octadecene 55a (0.071 g, 0.063 mmol) was reac-
ted by the procedure used to synthesize compound 33a
and afforded the title compound (0.059 g, 89%) as a
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-{3-
O-[4-(2,2-di-tert-butyloxycarbonylethyl)benzyl]-4,6-O-
benzylidene-ꢀ-D-galactopyranosyloxy}-4-octadecene 49b.
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-{3-O-
[4-(2,2-di-tert-butyloxycarbonylethyl)benzyl]-2-O-p-meth-
oxybenzyl-4,6-O-benzylidene-b-d-galactopyranosyloxy}-
4-octadecene 47b (0.124 g, 0.094 mmol) was reacted by
the procedure used to synthesize compound 48a and
afforded the title compound (0.080 g, 85%) as an oil.
1H NMR (400 MHz, CDCl3) d 0.89 (6H, t, J=6.7 Hz,
2Â -CH3), 1.25–1.42 (46H, m, -(CH2)11- and -(CH2)12-),
1.42 (18H, s, 2Â-OtBu), 1.58–1.61 (2H, m, -CH2-),
2.03 (2H, dt, J=6.9, 6.9 Hz,=CH-CH2-), 2.16 (2H,
m, -NHCOCH2-), 3.13 (2H, d, J=7.8 Hz, -CH2–
CH(CO2tBu)2), 3.35 (1H, s, H-50), 3.42–3.46 (2H, m, H-
30 and -CH-(CO2tBu)2), 3.82 (1H, dd, J=11.4, 3.8 Hz,
H-1), 3.96 (1H, dd, J=9.4, 7.8 Hz, H-20), 4.01 (1H, br d,
H-60), 4.08 (1H, d, J=3.3 Hz, H-40), 4.14 (1H, dd,
J=11.4, 5.5 Hz, H-1), 4.26 (1H, d, J=12.3 Hz, H-60),
4.31 (1H, d, J=7.7 Hz, H-10), 4.50 (1H, m, H-2), 4.73
(2H, s, -OCH2Ar), 5.45 (1H, s, -OCHO-), 4.51 (1H, dd,
J=15.3, 7.2 Hz, H-4), 5.61 (1H, br t, H-3), 5.87 (1H, dt,
J=15.3, 6.9 Hz, H-5), 6.29 (1H, d, J=9.1 Hz, -NH-),
7.17–7.19, 7.27–7.38, 7.41–7.45, 7.50–7.57 and 8.03–8.05
(14H, 5 sets of m, aromatic H).
1
white solid. H NMR (400 MHz, CD3OD) d 0.89 (6H,
t, J=6.8 Hz, 2Â-CH3), 1.26 (48H, m, -(CH2)11- and
-(CH2)13-), 2.06 (2H, m, ¼CH-CH2-), 2.20 (2H, m, -NH-
COCH2-), 3.14 (2H, d, J=7.7 Hz, -CH2-CH(CO2H)2),
3.58–3.69 (6H, 2 sets of m, H-1, -60, -50, H-30 and -CH-
(CO2H)2), 3.84 (1H, dd, J=10.9, 4.3 Hz, H-1), 3.92 (1H,
dd, J=10, 3.8 Hz, H-20), 4.03 (1H, d, J=2.8 Hz, H-40),
4.43–4.45 (1H, m, H-2), 4.60 and 4.69 (2Â1H, 2 d,
-OCH2Ar), 4.78 (1H, d, J=3.9 Hz, H-10), 5.53 (1H, dd,
J=15.1, 7.7 Hz, H-4), 5.61 (1H, br t, H-3), 5.88 (1H, dt,
J=15.1, 6.9 Hz, H-5), 7.21–7.23, 7.34–7.36, 7.45–7.49,
7.58–7.61 and 8.01–8.03 (9H, 4 sets of m, aromatic H).
1
Sodium salt: H NMR (400 MHz, CD3OD) d 0.89 (6H,
t, J=6.8 Hz, 2Â-CH3), 1.26–1.40 (46H, m, -(CH2)11-
and -(CH2)12-), 1.60 (2H, m, -CH2-), 2.06 (2H, dt,
J=6.8, 6.8 Hz, ¼CH-CH2-), 2.21 (2H, t, J=7.3 Hz,
-NHCOCH2-), 3.23–3.30 (2H, m, -CH2-CH(CO2Na)2),
3.60–3.77 (6H, m, H-1, H-60, H-30, H-50 and -CH-
(CO2Na)2), 3.83 (1H, dd, J=10.9, 4.1 Hz, H-1), 3.91
(1H, dd, J=10.0, 3.9 Hz, H-20), 4.01 (1H, d, J=2.7 Hz,
H-40), 4.44 (1H, m, H-2), 4.60 and 4.67 (2Â1H, 2 d,
J=11.5 Hz, -OCH2Ar), 4.77–4.79 (1H, d, J=3.9 Hz, H-
10), 5.53 (1H, dd, J=15.1, 7.9 Hz, H-4), 5.62 (1H, br t,
H-3), 5.89 (1H, dt, J=15.1, 6.8 Hz, H-5), 7.19–7.23,
7.30–7.33, 7.45–7.49, 7.58–7.62 and 8.02–8.04 (9H, 5
sets of m, aromatic H). IR (nujol) ꢀmax (cmÀ1) 3700–
3100 (O–H and N–H), 2920, 2860 (C–H), 1710, 1630
(C¼O).
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-{3-
O-[4-(2,2-di-tert-butyloxycarbonylethyl)benzyl]-b-D-galac-
topyranosyloxy}-4-octadecene 55b. (2S,3R,4E)-3-Benzoyl-
oxy-2-hexadecanoylamino-1-{3-O-[4-(2,2-di-tert-butyl-
oxycarbonylethyl)benzyl]-4,6-O-benzylidene-b-d-galac-
topyranosyloxy}-4-octadecene 49b (0.097 g, 0.08 mmol)
was reacted by the procedure used to synthesize com-
pound 40a and afforded the title compound (0.068 g,
76%) as an oil. 1H NMR (400 MHz, CDCl3) d 0.89 (6H,
t, J=6.7 Hz, 2Â-CH3), 1.25–1.42 (48H, m, -(CH2)11-
and -(CH2)13-), 1.42 (18H, s, 2Â-OtBu), 2.02–2.07 (2H,
m, ¼CH-CH2-), 2.10–2.25 (2H, m, -NHCOCH2-), 2.79,
2.85 and 3.06 (3Â1H, 3 s, 3Â-OH), 3.13 (2H, d,
J=7.9 Hz, -CH2-CH(CO2tBu)2), 3.37 (1H, dd, J=9.4,
3.3 Hz, H-30), 3.40 (1H, br dd, H-50), 3.45 (1H, t,
J=7.8 Hz, -CH(CO2tBu)2), 3.73 (1H, br d, H-60), 3.79
(1H, dd, J=9.4, 7.9 Hz, H-20), 3.83 (1H, dd, J=11.1,
4.3 Hz, H-1), 3.90 (1H, dd, J=11.8, 5.6Hz, H-60), 3.96
(1H, br d, H-40), 4.00 (1H, dd, J=11.1, 5.1 Hz, H-1), 4.28
(1H, d, J=7.7 Hz, H-10), 4.57 (1H, m, H-2), 4.72 (2H, s,
-OCH2Ar), 5.51 (1H, dd, J=15.3, 7.4 Hz, H-4), 5.65 (1H,
br t, H-3), 5.91 (1H, dt, J=15.3, 6.9Hz, H-5), 6.02 (1H, d,
J=9.3 Hz, -NH-), 7.20–7.22, 7.27–7.30, 7.44–7.48, 7.57–
7.61 and 8.03–8.05 (9H, 5 sets of m, aromatic H).
Synthesis of (2S,3R,4E)-3-benzoyloxy-2-hexadecanoyla-
mino-1-{3-O-[4-(2,2-di-carboxyethyl)benzyl]-ꢀ-D-galacto-
pyranosyloxy}-4-octadecene 56b
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-{3-
O-[4-(2,2-di-tert-butyloxycarbonylethyl)benzyl]-2-O-p-
methoxybenzyl-4,6-O-benzylidene-ꢀ-D-galactopyr-
anosyloxy}-4-octadecene 47b. (2S,3R,4E)-3-Benzoyloxy-
2-azido-1-{3-O-[4-(2,2-di-tert-butyloxycarbonylethyl)-
benzyl]-2-O-p-methoxybenzyl-4,6-O-benzylidene-b-d-
galactopyranosyloxy}-4-octadecene 46b (0.111 g, 0.10
mmol) was reacted by the procedure used to synthesize
compound 30a and afforded the title compound
1
(0.064 g, 65%) as a white solid. H NMR (400 MHz,
CDCl3) d 0.89 (6H, t, J=6.6 Hz, 2Â-CH3), 1.23–1.35
(46H, m, -(CH2)11- and -(CH2)12-), 1.41 (18H, s, 2Â
-OtBu), 1.41–1.51 (2H, m, -CH2-), 1.86 (2H, m,
-NHCOCH2-), 2.00 (2H, m, J=6.9 Hz, ¼CH-CH2-),
3.13 (2H, d, J=7.8 Hz, -CH2CH(CO2tBu)2), 3.29 (1H,
br s, H-50), 3.44 (1H, t, J=7.8 Hz, -CH(CO2tBu)2), 3.53
(1H, dd, J=9.5, 3.5 Hz, H-30), 3.70 (1H, dd, J=11.3,
4.0 Hz, H-1), 3.80 (3H, s, -OCH3), 3.84 (1H, br t, H-20),
4.00 (1H, d, J=11.6 Hz, H-60), 4.08 (1H, d, J=3.4 Hz,
H-40), 4.22 (1H, dd, J=11.3, 3.4 Hz, H-1), 4.24 (1H, d,
(2S,3R,4E)-3-Benzoyloxy-2-hexadecanoylamino-1-{3-
O-[4-(2,2-di-tert-butyloxycarbonylethyl)benzyl]-ꢀ-D-
galactopyranosyloxy}-4-octadecene 56b. (2S,3R,4E)-3-

A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
1423
Benzoyloxy-2-hexadecanoylamino-1-{3-O-[4-(2,2-di-tert-
butyloxycarbonylethyl)benzyl]-b-d-galactopyranosyloxy}-
4-octadecene 55b (0.068 g, 0.061 mmol) was reacted by
the procedure used to synthesize compound 33a and
afforded the title compound (0.067 g, quant.) as a white
foam. Sodium salt: 1H NMR (400 MHz, CD3OD) d
0.89 (6H, t, J=6.8 Hz, 2Â-CH3), 1.26–1.39 (46H, m,
-(CH2)11- and -CH2)12-), 1.59 (2H, m, -CH2-), 2.06 (2H,
dt, J=6.8, 6.8 Hz, ¼CH-CH2-), 2.19 (2H, m, -NH-
COCH2-), 3.14 (2H, d, J=7.7 Hz, -CH2CH(CO2H)2),
3.34 (1H, dd, J=9.7, 3.2 Hz, H-30), 3.40 (1H, br dd, H-
50), 3.58–3.71 (5H, m, -CH(CO2H)2, H-1, H-60 and H-
20), 3.98 (1H, d, J=2.9 Hz, H-40), 4.10 (1H, dd, J=10.4,
5.3 Hz, H-60), 4.22 (1H, d, J=7.7 Hz, H-10), 4.44 (1H, m,
H-2), 4.61 and 4.72 (2Â1H, 2 d, J=11.7 Hz, -OCH2Ar),
5.51 (1H, dd, J=14.9, 7.8 Hz, H-4), 5.57 (1H, br t, H-3),
5.89 (1H, dt, J=14.9, 6.8 Hz, H-5), 7.20–7.22, 7.35–
7.37, 7.43–7.48, 7.57–7.61 and 7.99–8.02 (9H, 5 sets of
m, aromatic H). IR (nujol) ꢀmax (cmÀ1) 3700–3100 (O–
H and N–H), 2920, 2860 (C–H), 1715, 1630 (C¼O).
7.49, 7.53–7.59 and 8.08–8.14 (10H, 3 sets of m, aro-
matic H). IR (film) ꢀmax (cmÀ1) 2980, 2930, 2860 (C–H),
.
1730 (C¼O). Anal. calcd for C43H60O13 0.1CH2Cl2: C,
65.24; H, 7.65. Found: C, 65.01; H, 7.49.
Methyl 9-[2-O-(3,3-di-carboxypropyl)-3-O-benzoyl-a-D-
galactopyranosyloxy]-nonanoate 60a. Anisole (0.30 mL)
was added to neat methyl 9-[2-O-(3,3-di-tert-butyloxy-
carbonylpropyl)-3-O-benzoyl-4,6-O-benzylidene-a-d-gal-
actopyranosyloxy]-nonanoate 59a (0.412 g, 0.52 mmol)
at 0 ꢁC. 90% Aqueous trifluoroacetic acid (15 mL) was
then added and the mixture was stirred at 0 ꢁC for 5
min, then toluene (15 mL) was added. The solvents were
evaporated and the co-evaporation was repeated twice.
The same procedure was repeated again and the residue
was purified by silica gel chromatography (3Â6 cm, 2.5–
30% methanol/chloroform +5% water) and afforded
the title compound (0.175 g, 58%) as a solid. MS (ESIÀ)
583.2 (M–H)À. MS (ESI+) 585.4 (MH)+, 607.2
(M+Na)+. HRMS (ESIÀ) calcd for C28H39O13:
1
583.23905. Found: 583.2399 d=À1.4 ppm. Diacid: H
Synthesis of methyl 9-[2-O-(3,3-di-carboxypropyl)-3-O-
benzoyl-ꢁ-D-galactopyranosyloxy]-nonanoate 60a
NMR (400 MHz, pyridine-d5) d 1.17–1.21, 1.28–1.34,
1.53–1.68 (6H, 2H and 4H, -(CH2)6-), 2.28 (2H,
t, J=7.4 Hz, -CH2CO2Me), 2.69 (2H, dt, J=6.5, 6.5 Hz,
-CH2CH(CO2H)2), 3.58 (3H, s, -OCH3), 3.65 (1H,
dt, J=9.3, 6.6 Hz, -OCH2-), 3.94 (1H, dt, J=9.3,
6.8 Hz, -OCH2-), 4.18 (2H, t, J=6.1 Hz, -OCH2-), 4.29
(1H, t, J=7.3 Hz, -CH(CO2H)2), 4.41 (1H, dd, J=10.6,
6.2 Hz, H-60), 4.46 (1H, dd, J=10.6, 6.2 Hz, H-60), 4.52
(1H, br dd, J=6.0, 6.0 Hz, H-50), 4.80 (1H, dd, J=10.4,
3.5 Hz, H-20), 5.08 (1H, d, J=2.8 Hz, H-40), 5.58 (1H, d,
J=3.5 Hz, H-10), 6.03 (1H, dd, J=10.4, 2.8 Hz, H-30),
7.29–7.40 and 8.26–8.28 (5H, 2 sets of m, aromatic H).
Sodium salt: 1H NMR (400 MHz, CD3OD) d 1.29
and 1.55–1.61 (8H and 4H, -(CH2)6-), 2.03 (2H, m,
-CH2CH(CO2Na)2), 2.26 (2H, br t, -CH2CO2Me),
3.04 (1H, t, J=6.5 Hz, -CH(CO2Na)2), 3.46 (1H, br dt,
-OCH2-), 3.53–3.64 (4H, m, H-60 and -OCH2-), 3.58
(3H, s, -OCH3), 3.60 (1H, br dt, -OCH2-), 3.85 (1H, dd,
J=5.6, 5.6 Hz, H-50), 3.99 (1H, br d, J=10.5 Hz, H-20),
4.13 (1H, br s, H-40), 5.03 (1H, br s, H-10), 5.17 (1H, br
d, J=10.5 Hz, H-30), 7.40–7.44, 7.51–7.54 and 8.01–8.03
Methyl 9-[2-O-(3,3-di-tert-butyloxycarbonylpropyl)-3-
O-benzoyl-4,6-O-benzylidene-ꢁ-D-galactopyranosyloxy]-
nonanoate 59a and methyl 9-[2-O-(3,3-di-tert-butylox-
ycarbonylpropyl)-4,6-O-benzylidene-3-O-benzoyl-ꢀ-D-
galactopyranosyloxy]-nonanoate 59b. Ethyl 3-O-benzoyl-
4,6-O-benzylidene-2-O-(3,3-di-tert-butyloxycarbonyl-
propyl)-1-thio-b-d-galactopyranoside 20 (0.800 g, 1.21
mmol), 2,6-di-tert-butyl-4-methylpyridine (0.500 g,
2.42 mmol) and methyl 9-hydroxynonanoate 58
(0.228 g, 1.21 mmol) were reacted by the procedure used
to synthesize compounds 29a and b except that dioxane
was used as solvent. This afforded the a-anomer
(0.503 g, 53%) and the b-anomer (0.528 g, 56%) of the
22
title material as oils. ꢁ-Anomer: ½aꢂD +112ꢁ (c 1.1,
1
CHCl3). H NMR (400 MHz, CDCl3) d 1.33–1.43 and
1.62–1.68 (8H and 4H, -(CH2)6-), 1.40 and 1.41 (2Â9H,
2s, 2Â-OtBu), 2.04 (2H, m, -CH2CH(CO2tBu)2), 2.32
(2H, t, J=7.5 Hz, -CH2CO2Me), 3.38 (1H, t, J=7.5 Hz,
-CH(CO2tBu)2), 3.57 (1H, dt, J=9.7, 6.6 Hz, -OCH2-),
3.66–3.68 (2H, m, -OCH2-), 3.68 (3H, s, -OMe), 3.75
(1H, dt, J=9.7, 7.0 Hz, -OCH2-), 3.84 (1H, s, H-5),
4.08–4.12 (2H, m, H-60 and H-20), 4.30 (1H, dd, J=12.4,
1.0 Hz, H-60), 3.62 (1H, d, J=3.3 Hz, H-40), 5.13 (1H, d,
J=3.4 Hz, H-10), 5.46 (1H, dd, J=10.5, 3.6 Hz, H-30),
3.52 (1H, s, -OCHO-), 7.32–7.34, 7.43–7.49, 7.53–7.59
and 8.10–8.12 (10H, 4 sets of m, aromatic H). Anal.
(5H, 3 sets of m, aromatic H). IR (KBr) ꢀmax (cmÀ1
)
3700–3000 (O–H), 3030 and 2960 (C–H), 1720 (C¼O)
and 1580.
Synthesis of methyl 9-[2-O-(3,3-di-carboxypropyl)-3-O-
benzoyl-ꢀ-D-galactopyranosyloxy]-nonanoate 60b. Methyl
9-[2-O-(3,3-di-tert-butyloxycarbonylpropyl)-3-O-benzoyl-
4,6-O-benzylidene-a-d-galactopyranosyloxy]-nonanoate
59b (0.52 g, 0.66 mol) was reacted by the general proce-
dure used to synthesize compound 33a and afforded the
title compound (0.191 g, 50%) as a white foam. Diacid:
1H NMR (400 MHz, pyridine-d5) d 1.28–1.32, 1.52–1.57
and 1.66–1.73 (8H, 2H and 2H, 3 sets of m, -(CH2)6-),
2.28 (2H, t, J=7.5 Hz, -CH2CO2Me), 2.74 (2H, m,
-CH2CH(CO2H)2), 3.61 (3H, s, -OCH3), 3.69 (1H,
dt, J=9.2, 6.9 Hz, -OCH2-), 4.06 (1H, dt, J=9.2,
6.9 Hz, -OCH2-), 4.10 (1H, dd, J=6.1, 6.1 Hz, H-50),
4.22–4.30 and 4.38–4.51 (4H and 2H, 2 sets of m, H-60,
-OCH2-, H-20 and -CH(CO2H)2), 4.83 (1H, d,
J=7.6 Hz, H-10), 4.96 (1H, d, J=3.0 Hz, H-40), 5.60
(1H, dd, J=10.1, 3.1 Hz, H-30), 7.29–7.39 and 8.25–8.31
.
calcd for C43H60O13 0.1CH2Cl2: C, 65.24; H, 7.65.
Found: C, 65.10; H, 7.48. ꢀ-Anomer: [a]2D2 +54ꢁ (c 1.0,
1
CHCl3). H NMR (400 MHz, CDCl3) d 1.22–1.38 and
1.64 (8H and 4H, 2 sets of m, -(CH2)6-), 1.38 (18H, s,
2Â -OtBu), 2.00 (2H, m, -CH2CH(CO2tBu)2), 2.31
(2H, t, J=7.5 Hz, -CH2CO2Me), 3.31 (1H, t, J=7.4 Hz,
-CH(CO2tBu)2), 3.44–3.56 (1H, m, -OCH2-), 3.54 (1H,
s, H-50), 3.62–3.68 (1H, m, -OCH2-), 3.68 (3H, s,
-OCH3), 3.80 (1H, dd, J=10.1, 7.7 Hz, H-20), 3.90 (1H,
dt, J=10.1, 6.0 Hz, -OCH2-), 3.97 (1H, dt, J=9.1,
6.9 Hz, -OCH2-), 4.08 (1H, d, J=12.1 Hz, H-60), 4.35
(1H, d, J=12.1 Hz, H-60), 4.44 (1H, d, J=7.7 Hz, H-10),
4.49 (1H, d, J=3.6 Hz, H-40), 5.08 (1H, dd, J=10.1,
3.6 Hz, H-30), 5.50 (1H, s, -OCHO-), 7.32–7.36, 7.40–

1424
A. Marinier et al. / Bioorg. Med. Chem. 9 (2001) 1395–1427
(5H, 2 sets of m, aromatic H). Anal. calcd for
C28H40O13 0.25 H2O: C, 56.48; H, 6.86. Found: C,
J=7.5 Hz, -CH2CO2Me), 3.23 (1H, br dd, H-20), 3.43
(1H, t, J=7.3 Hz, -CH(CO2tBu)2), 3.49 (1H, dt, J=9.3,
6.9 Hz, -OCH2-), 3.67 (3H, s, -OCH3), 3.71 (1H, dt,
J=9.9, 6.1 Hz, -OCH2-), 3.78–3.85 (3H, m, H-50, H-60
and -OCH2-), 3.87 (1H, dt, J=9.3, 6.9 Hz, -OCH2-),
3.94–4.00 (1H, m, H-60), 4.09 (1H, dd, J=6.4, 5.9 Hz,
H-30), 4.13 (1H, dd, J=5.8, 1.2 Hz, H-40), 4.23 (1H, d,
J=7.9 Hz, H-10). IR (film) ꢀmax (cmÀ1) 3700–3200 (O–
H), 2980, 2935, 2860 (C–H), 1740, 1725 (C¼O).
.
56.42; H, 6.74. MS (ESIÀ) 583.3 (MÀH)À. Sodium salt:
1H NMR (400 MHz, CD3OD) d 1.26–1.33 and 1.52–
1.59 (8H and 4H, -(CH2)6-), 1.91–2.06 (2H, m, -CH2-
CH(CO2Na)2), 2.25 (2H, t, J=7.4 Hz, -CH2CO2Me),
2.99 (1H, t, J=7.4 Hz, -CH(CO2Na)2), 3.50–3.67 and
3.80–3.86 (6H and 2H, 2Â-OCH2-, H-20, H-40, H-50 and
H-60), 3.58 (3H, s, -OCH3), 4.08 (1H, d, J=3.2 Hz, H-
40), 4.35 (1H, d, J=7.7 Hz, H-10), 4.85 (1H, dd, J=10.1,
3.3 Hz, H-30), 7.40–7.44, 7.50–7.54 and 8.01–8.03 (5H, 3
sets of m, aromatic H). IR (KBr) ꢀmax (cmÀ1) 3700–3000
(O-H), 2930, 2850 (C–H), 1720 (C¼O) and 1580.
Methyl 9-[2-O-(3,3-dicarboxypropyl)-ꢁ-D-galactopyr-
anosyloxy]-nonanoate 62a. Methyl 9-[2-O-(3,3-di-tert-
butyloxycarbonylpropyl)-3,4-O-isopropylidene-b-d-gal-
actopyranosyloxy]-nonanoate 61a (0.291 g, 0.46 mmol)
was reacted by the procedure used to synthesize com-
pound 33a and afforded the title compound (0.095 g,
Synthesis of methyl 9-[2-O-(3,3-di-carboxypropyl)-ꢁ-D-
galactopyranosyloxy]-nonanoate 62a
Methyl
9-[2-O-(3,3-di-tert-butyloxycarbonylpropyl)-
1
3,4-O-isopropylidene-ꢁ-D-galactopyranosyloxy]-non-
anoate 61a and methyl 9-[2-O-(3,3-di-tert-butyloxy-
carbonylpropyl)-3,4-O-isopropylidene-ꢀ-D-galactopyr-
anosyloxy]-nonanoate 61b. Ethyl 3,4-O-isopropylidene-6
-O-tert-butyldimethylsilyl-2-O-(3,3-di-tert-butyloxycar-
bonylpropyl)-1-thio-b-d-galactopyranoside 12 (0.900 g,
1.45 mmol), 2,6-di-tert-butyl-4-methylpyridine (0.600 g,
2.90 mmol), methyl 9-hydroxynonanoate 58 (0.273 g,
1.45 mmol) were reacted by the procedure used to syn-
thesize compounds 29a and b and afforded a mixture of
a- and b-anomers of the title material (0.900 g, 83%) as
a yellow oil. This mixture was dissolved in dry tetra-
hydrofuran (10 mL) and was treated with acetic acid
(0.4 mL) and tetrabutylammonium fluoride (1 M in tet-
rahydrofuran, 5 mL, 5 mmol) at 0 ꢁC. The mixture was
stirred 45 min at 0 ꢁC and 1 h at 23 ꢁC. Tetra-
butylammonium fluoride (2.5 mL) was added again and
the mixture was stirred for two additional h at 23 ꢁC
and stored at À20 ꢁC overnight. The mixture was dilu-
ted with toluene and co-evaporated (2Â). The residue
was dissolved in ethyl acetate (100 mL) and washed with
aqueous saturated sodium bicarbonate (2Â50 mL),
water (50 mL) and brine, dried over anhydrous sodium
sulfate, filtered and concentrated. The residue was pur-
ified by silica gel chromatography (3.5Â14 cm, 30–50%
ethyl acetate/hexane) and afforded the a-anomer
(0.306 g, 40%) and the b-anomer (0.280 g, 37%) of the
title compound as clear oils. ꢁ-Anomer: [a]2D2 +60ꢁ (c
43%) as a yellow oil. Diacid: H NMR (400 MHz, pyri-
dine-d5) d 1.16, 1.28–1.34, 1.52–1.61, 1.63–1.71 (6H, 2H,
2H and 2H, 4 sets of m, -(CH2)6-), 2.27 (2H, t,
J=7.5 Hz, -CH2CO2Me), 2.72 (2H, dt, J=6.4, 6.4 Hz,
-CH2CH(CO2H)2), 3.60 (3H, s, -OCH3), 3.60 (1H,
dt, J=9.5, 6.5 Hz, -OCH2-), 3.94 (1H, dt, J=9.3,
6.7 Hz, -OCH2-), 4.08–4.18, 4.31–4.45 (2H and 5H, 2
sets of m, -OCH2-, H-60, H-50, H-20 and -CH(CO2H)2),
4.58 (1H, dd, J=9.7, 3.2 Hz, H-30), 4.61 (1H, d,
J=3.2 Hz, H-40), 5.46 (1H, d, J=3.5 Hz, H-10). Sodium
1
salt: H NMR (400 MHz, CD3OD) d 1.27 and 1.55 (8H
and 4H, 2 sets of m, -(CH2)6-), 2.03–2.08 (2H, m, -CH2-
CH(CO2Na)2), 2.26 (2H, t, J=7.4 Hz, -CH2-CO2Me),
3.20 (1H, t, J=7.4 Hz, -CH(CO2Na)2), 3.39 (1H, dt,
J=9.5, 6.4 Hz, -OCH2-), 3.48 (1H, dt, J=9.2, 7.3 Hz,
-OCH2-), 3.54 (1H, dd, J=10.0, 3.5 Hz, H-20), 3.59 (3H,
s, -OCH3), 3.62–3.73 (5H, m, -OCH2-, H-60 and H-50),
3.75 (1H, dd, J=10.0, 3.2 Hz, H-30), 3.83 (1H, br d,
J=2.6 Hz, H-40), 4.92 (1H, d, H-10). IR (KBr) ꢀmax
(cmÀ1) 3800–3000 (O–H), 2940 (C–H), 1740 (C¼O) and
1580.
Synthesis of methyl 9-[2-O-(3,3-di-carboxypropyl)-ꢀ-D-
galactopyranosyloxy]-nonanoate 62b. A solution of
methyl 9-[2-O-(3,3-di-tert-butyloxycarbonylpropyl)-3,4-
O-isopropylidene-b-d-galactopyranosyloxy]-nonanoate
61b (0.207 g, 0.33 mmol) was reacted by the procedure
used to synthesize compound 33a and gave the title
material (0.100 g, 63%) as a white solid. Diacid: 1H
NMR (400 MHz, pyridine-d5) d 1.15, 1.29–1.32, 1.51–
1.57 and 1.64–1.71 (6H and 3Â2H, 4 sets of m, -(CH2)6-),
2.28 (2H, t, J=7.5 Hz, -CH2CO2Me), 2.78 (2H, m,
-CH2CH(CO2H)2), 3.61 (3H, s, -OCH3), 3.66, 4.03–4.11
and 4.50–4.55 (1H, 3H and 1H, 3 sets of m, -OCH2-,
H-50, H-40 and H-30), 3.97 (1H, t, J=5.9 Hz,
-CH(CO2H)2), 4.21 (1H, dt, J=9.6, 6.0 Hz, -OCH2-),
4.39 (1H, br t, H-20), 4.42 (2H, d, J=6.1 Hz, H-60), 4.53
(1H, dt, J=9.9, 6.0 Hz, -OCH2-), 4.71 (1H, d,
J=6.7 Hz, H-10). Sodium salt: 1H NMR (400 MHz,
CD3OD) d 1.27 and 1.53–1.55 (8H and 4H, 2 sets of
m, -(CH2)6-), 1.98–2.10 (2H, m, -CH2CH(CO2Na)2),
2.26 (2H, t, J=7.4 Hz, -CH2CO2Me), 3.18–3.26, 3.44–
3.54, 3.82–3.86 and 3.95–3.98 (2H, 3H, 1H and 1H, 4
sets of m, 2Â-OCH2-, H-20, H-30 and H-50), 3.39 (1H, t,
J=5.9 Hz, -CH(CO2Na)2), 3.59 (3H, s, -OCH3), 3.67
(2H, d, J=6.1 Hz, H-60), 3.78 (1H, d, J=2.7 Hz, H-40),
1
1.3, CHCl3). H NMR (400 MHz, CDCl3) d 1.31–1.35
and 1.61–1.62 (8H and 4H, 2 sets of m, -(CH2)6-),
1.35 and 1.52 (2Â3H, 2 s, -C(CH3)2-), 1.46 (18H, s,
2Â-OtBu), 2.04–2.15 (2H, m, -CH2-CH(CO2tBu)2), 2.31
(2H, t, J=7.5 Hz, -CH2CO2Me), 3.41 (1H, t, J=7.4 Hz,
-CH(CO2tBu)2), 3.42 (1H, dd, J=7.7, 3.4 Hz, H-20),
3.47 (1H, dt, J=9.7, 6.6 Hz, -OCH2-), 3.58–3.70 (3H, m,
H-50 and -OCH2-), 3.68 (3H, s, -OCH3), 3.74 (1H, dt,
J=9.9, 6.0 Hz, -OCH2-), 3.96 (1H, dd, J=11.7, 6.1 Hz,
H-60), 4.04–4.07 (1H, br d, H-60), 4.23 (1H, dd, J=5.6,
2.6 Hz, H-40), 4.28 (1H, dd, J=7.6, 5.7 Hz, H-30), 4.91
(1H, d, J=3.4 Hz, H-10). IR (film) ꢀmax (cmÀ1) 3650–
3200 (O-H), 2980, 2930, 2860 (C-H), 1740, 1725 (C¼O).
ꢀ-Anomer: [a]2D2 +10ꢁ (c 1.3, CHCl3). 1H NMR
(400 MHz, CDCl3) d 1.27–1.31 and 1.61–1.62 (8H
and 4H, 2 sets of m, -(CH2)6-), 1.33 and 1.51 (2Â3H, 2 s,
-C(CH3)2-), 1.46 and 1.47 (2Â9H, 2 s, 2Â-OtBu), 2.05–
2.11 (2H, m, -CH2CH(CO2tBu)2), 2.31 (2H, t,