FGF RECEPTOR-ACTIVATING N-ACYL OCTASACCHARIDES,
PREPARATION THEREOF, AND THERAPEUTIC USE THEREOF
The present invention relates to N-acyl octasaccharides that are agonists of the
FGF/FGFR system, and to their preparation and therapeutic use.
Angiogenesis is a process of generation of new blood capillaries. During the
blockage of a blood vessel, angiogenesis, combined with arteriogenesis (dilation of the
capillaries), improves the revascularization of the blocked area. It has been shown in
vitro and in vivo that several growth factors, such as Fibroblast Growth Factors (FGFs),
stimulate the neovascularization process.
FGFs are a family of 23 members. FGF2 (or basic FGF) is an 18 kDa protein.
FGF2 induces, in endothelial cells in culture, their proliferation and migration and the
production of proteases, in vivo, FGF2 promotes neovascularization. FGF2 interacts
with endothelial cells via two classes of receptors, the high-affinity receptors with
tyrosine kinase activity (FGFRs) and the low-affinity receptors of heparan sulfate
proteoglycan (HSPG) type.
It is known that cell surface receptors with tyrosine kinase activity associate in
dimeric form with a complex formed from two ligand molecules and one heparan sulfate
molecule. The formation of this complex triggers a cascade of intracellular signals
resulting in activation of cell proliferation and migration, which are two key processes
involved in angiogenesis.
Thus, FGF2 and its receptors represent very pertinent targets for therapies
directed towards activating or inhibiting angiogenesis processes.
Synthetic oligosaccharides have also been the subject of studies of interactions
with the FGF receptors (C. Tabeur et al., Bioorg. & Med. Chem., 1999, 7, 2003-2012;
C. Noti et al., Chem. Eur. J., 2006, 12, 8664-8686).
We have now found novel synthetic oligosaccharides that are capable of
facilitating the formation of the FGF/FGFR complex and of thus promoting the in vitro
survival of endothelial cells and of increasing the in vitro and in vivo formation of new
blood vessels.
One subject of the present invention is novel octasaccharide compounds
corresponding to formula (I):

in which:
- the wavy line denotes a bond located either below or above the plane of the
pyranose ring of the glucosamine unit,
- R1 represents a group O-alkyl, in which the said alkyl group comprises from 1
to 16 carbon atoms and is optionally substituted with one or more (for example 1 or 2)
groups, which may be identical or different, chosen from aryl and cycloalkyl groups,
- R2 represents either a group OSO3" or a hydroxyl group,
- R3 represents an alkyl, cycloalkyl or alkyl-cycloalkyl group, and
- R4 represents a disaccharide of formula (II):
in which:
- R3 is as defined above,
- R5 represents either a group OSO3" or a hydroxyl group, and
- R6 represents a disaccharide of formula (III):

in which:
- R3 is as defined above,
- R7 represents either a group OSO3" or a hydroxyl group, and
- R8 represents a disaccharide of formula (IV):

in which:
- R3 is as defined above,
- R9 represents either a group OSO3" or a hydroxyl group.
In the context of the present invention, and unless otherwise mentioned in the
text, the following definitions apply:
- alkyl group: a linear or branched saturated aliphatic group. Unless otherwise
indicated in the text, such an alkyl group advantageously comprises between 1 and 6
carbon atoms. Examples that may be mentioned include methyl, ethyl, propyl, isopropyl,
butyl, isobutyl, tert-butyl, pentyl, etc. groups;
- cycloalkyl group: a cyclic alkyl group comprising from 3 to 6 carbon atoms, for
example a cyclopentyl or cyclohexyl group;
- alkyl-cycloalkyl group: an alkyl group which is itself substituted with a
cycloalkyl group, the said alkyl and cycloalkyl groups being as defined above; and
- aryl group: a cyclic aromatic group comprising between 5 and 10 carbon
atoms, such as a phenyl group. Such an aryl group is optionally substituted with one or
more groups such as halogen atoms and alkyl, alkoxy, thioalkyl, trifluoromethyl and
phenyl groups.
The octasaccharides according to the invention are synthetic octasaccharides,
i.e. they are compounds obtained by total synthesis starting from intermediate synthons,
as will be described in detail in the text hereinbelow. In this respect, they differ from
oligosaccharides obtained by depolymerization or isolation from complex mixtures of
polysaccharides, such as heparins or low molecular weight heparins. In particular, the
compounds according to the invention have a well-defined structure resulting from their
chemical synthesis and are in the form of pure octasaccharides, i.e. they are free of
other octasaccharide species.
The invention encompasses the compounds of formula (I) in acid form or in the
form of any pharmaceutically acceptable salt thereof. In the acid form, the functions
-COO" and -SO3" are, respectively, in -COOH and -SO3H form.
The term "pharmaceutically acceptable salt of the compounds of the invention"
means a compound in which one or more of the functions -COO" and/or -SO3" are
ionically linked to a pharmaceutically acceptable cation. The preferred salts according to
the invention are those in which the cation is chosen from alkali metal cations,
especially the Na+ cation.
The compounds of formula (I) according to the invention also comprise those in
which one or more hydrogen or carbon atoms have been replaced with a radioactive
isotope, for example tritium or carbon 14C. Such labelled compounds are useful in
research, metabolism or pharmacokinetic studies, as ligands in biochemical tests.
In formula (I) of the compounds according to the present invention, it is
understood that:
- the disaccharide of formula (II) is linked to the disaccharide unit represented in
formula (I) via the oxygen atom located in position 1 of its glucosamine unit,
- similarly, the disaccharide of formula (III) is linked to the disaccharide of
formula (II) via the oxygen atom located in position 1 of its glucosamine unit,
- similarly, the disaccharide of formula (IV) is linked to the disaccharide of
formula (III) via the oxygen atom located in position 1 of its glucosamine unit.
The term "glucosamine unit" means the monosaccharide unit having the
following formula:

The other type of saccharide unit present in the compounds according to the
invention is a uronic acid, more specifically an iduronic acid, corresponding to the
following formula:

Thus, the compounds of formula (I) according to the invention, which are
octasaccharides, may also be represented according to formula (I') as follows, in which
the iduronic units and the glucosamine units succeed each other and in which R1, R2,
R3, R5, R7 and R9 are as defined previously:
Among the compounds of formula (l)/(l') that are subjects of the invention,
mention may be made of a subgroup of compounds in which R1 represents:
- either a group O-alkyl, in which the said alkyl group comprises from 5 to 16
carbon atoms and advantageously from 5 to 12 carbon atoms (for example a group -O-
pentyl, -O-undecyl or 2-propylpentyl),
- or a group O-alkyl, in which the said alkyl group comprises from 1 to 8 carbon
atoms and advantageously from 3 to 6 carbon atoms (for example a group -O-propyl or
-O-pentyl), and is substituted with 1 or 2 groups, which may be identical or different,
chosen from aryl and cycloalkyl groups (for example phenyl and cycloalkyl groups, such
as cyclohexyl).
Among the compounds of formula (l)/(l') that are subjects of the invention,
mention may be made of another subgroup of compounds, in which R3 represents either
an alkyl group comprising from 2 to 6 carbon atoms (for example a propyl or pentyl
group) or a cycloalkyl group (for example a cyclopentyl group).
Among the compounds of formula (l)/(l') that are subjects of the invention,
mention may be made of another subgroup of compounds, in which at least one of the
groups R2, R5, R7 and R9 represents a hydroxyl group.
Another subgroup of compounds that are subjects of the invention is such that at
least one of the groups R2, R5, R7 and R9 represents a hydroxyl group and at least one
of the groups R2, R5, R7 and R9 represents a group OSO3".
Other subgroups of octasaccharides according to the invention may have
several of the characteristics listed above for each of the subgroups defined previously.
Mention may be made in particular of the subgroup of compounds of formula (I)
in which:
- R1 represents:
. either a group O-alkyl, in which the said alkyl group comprises from 5 to
16 carbon atoms and advantageously from 5 to 12 carbon atoms (for example a group
-O-pentyl, -O-undecyl or 2-propylpentyl),
. or a group O-alkyl, in which the said alkyl group comprises from 1 to 8
carbon atoms and advantageously from 3 to 6 carbon atoms (for example a group -O-
propyl or -O-pentyl), and is substituted with 1 or 2 groups, which may be identical or
different, chosen from aryl and cycloalkyl groups (for example phenyl and cycloalkyl
groups, such as cyclohexyl),
- R3 represents either an alkyl group comprising from 2 to 6 carbon atoms (for
example a propyl or pentyl group) or a cycloalkyl group (for example a cyclopentyl
group), and
- at least one of the groups R2, R5, R7 and R9 represents a hydroxyl group.
Among the compounds of the invention, mention may be made especially of the
following octasaccharides:
- Pentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium
sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-
(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-acetamido-2-
deoxy-6-O-sodium sulfonato-(3--D-glucopyranoside (No. 1);
- Pentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
deoxy-2-(1 -oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium
2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1 -oxobutyl)amino-6-
O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopyranosyluronate)-(1?4)-(2-deoxy-2-(1 -oxobutyl)amino-6-O-sodium sulfonato-a-D-
glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-
2-deoxy-2-(1-oxobutyl)amino-6-O-sodium sulfonato-ß--D-glucopyranoside (No. 2);
- Pentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
deoxy-2-(1 -oxohexyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium
2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1 ^4)-(2-deoxy-2-(1 -oxohexyl)amino-6-
O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopyranosyluronate)-(1?4)-(2-deoxy-2-(1-oxohexyl)amino-6-O-sodium sulfonato-a-D-
glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-
2-deoxy-2-(1-oxohexyl)amino-6-O-sodium sulfonato-(3--D-glucopyranoside (No. 3);
- Pentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
[(cyclopentylcarbonyl)amino]-2-deoxy-6-O-sodium sulfonato-a-D-glucopyrariosyl)-(1?4)-
(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-[(cyclopentyl-
carbonyl)amino]-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-
O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-[(cyclopentylcarbonyl)amino]-2-
deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-
a-L-idopyranosyluronate)-(1?4)-2-[(cyclopentylcarbonyl)amino]-2-deoxy-6-O-sodium
sulfonato-ß--D-glucopyranoside (No. 4);
- Undecyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium
sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-
(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-acetamido-2-
deoxy-6-O-sodium sulfonato-(3--D-glucopyranoside (No. 5);
- 5-phenylpentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-
(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium
sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-
(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-acetamido-2-
deoxy-6-O-sodium sulfonato-ß--D-glucopyranoside (No. 6);
- Methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-[(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium
sulfonato-a-D-glucopyranosyl)]2-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronate)-(1?4)-2-acetamido-2-deoxy-6-O-sodium suIfonato-a-D-glucopyranoside (No.
7);
- Methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4))-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4))-(2-acetamido-2-deoxy-6-O-sodium
sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronate)-(1?4)-(2-acetamido-2-deoxy-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium
sulfonato-a-L-idopyranosyluronate)-(1?4)-2-acetamido-2-deoxy-6-O-sodium sulfonato-
a-D-glucopyranoside (No. 8);
- Methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium
sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronate)-(1?4)-(2-acetamido-2-deoxy-a-D-glucopyranosyl)-(1?4))-(sodium 2-O-sodium
sulfonato-a-L-idopyranosyluronate)-(1?4)-2-acetamido-2-deoxy-6-O-sodium sulfonato-
-a-D-glucopyranoside (No. 9);
- Methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-gluco-
pyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-acetamido-2-deoxy-a-D-gluco-
pyranoside (No. 10);
- Methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-gluco-
pyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-acetamido-2-deoxy-6-O-sodium
sulfonato-a-D-glucopyranoside (No. 11);
- Methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-a-D-gluco-
pyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4))-2-acetamido-2-deoxy-a-D-gluco-
pyranoside (No. 12);
- Methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-a-D-gluco-
pyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-acetamido-2-deoxy-6-O-sodium
sulfonato-a-D-glucopyranoside (No. 13);
- 5-phenylpentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-
(2-deoxy-2-(1-oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-
(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1-
oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium
sulfonato-a-L-idopyranosyluronate)-(1?4))-(2-deoxy-2-(1-oxobutyl)amino-a-D-gluco-
pyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-
deoxy-2-(1-oxobutyl)amino-ß--D-glucopyranoside (No. 14);
- 2-propylpentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-
(2-deoxy-2-(1 -oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranpsyl)-(1?4)-
(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1 -
oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium
sulfonato-a-L-idopyranosyluronate)-(1?4))-(2-deoxy-2-(1-oxobutyl)amino-a-D-gluco-
pyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-
deoxy-2-(1-oxobutyl)amino-(3--D-glucopyranoside (No. 15);
- 3-cyclohexylpropyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-
(1?4)-(2-deoxy-2-acetamido-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium
2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1-44)-(2-deoxy-2-acetamido-6-O-
sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopyranosyluronate)-(1?4)-(2-deoxy-2-acetamido-a-D-glucopyranosyl)-(1?4)-(sodium
2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-deoxy-2-acetamido-ß--D-gluco-
pyranoside (No. 16);
- 3-cyclohexylpropyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-
(1?4)-(2-deoxy-2-(1 -oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-
(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1 -
oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium
sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1-oxobutyl)amino-a-D-gluco-
pyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-
deoxy-2-(1-oxobutyl)amino-ß--D-glucopyranoside (No. 17);
- 3,3-diphenylpropyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-
(1?4)-(2-deoxy-2-(1 -oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-
(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1 -
oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium
sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1-oxobutyl)amino-a-D-gluco-
pyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-
deoxy-2-(1-oxobutyl)amino-ß-D-glucopyranoside (No. 18); and
- 5-phenylpentyl (sodium 2-O-sodium sulfonato-a-L_-idopyranosyluronate)-(1?4)-
(2-deoxy-2-(3-methyl-1-oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-
(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(3-
methyl-1-oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-
O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(3-methyl-1-
oxobutyl)amino-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopyranosyluronate)-(1?4))-2-deoxy-2-(3-methyl-1-oxobutyl)amino-P--D-gluco-
pyranoside (No. 19).
In its principle, the process for preparing the compounds according to the
invention uses di- or oligosaccharide synthons prepared as reported previously in the
literature. Reference will be made especially to the patents or patent applications
EP 0 300 099, EP 0 529 715, EP 0 621282 and EP 0 649 854, and also to the
publication by C. Van Boeckel and M. Petitou published in Angew. Chem. Int. Ed. Engl.,
1993, 32, 1671-1690. These synthons are then coupled together so as to give an
entirely protected equivalent of an octasaccharide according to the invention. This
protected equivalent is then converted into a compound according to the invention. In
the coupling reactions mentioned above, a "donor" di- or oligosaccharide, activated on
its anomeric carbon, reacts with an "acceptor" di- or oligosaccharide, bearing a free
hydroxyl.
The specific synthetic schemes will be described in the detailed examples that
follow.
The present invention thus relates to a process for preparing octasaccharides of
formula (l)/(l'), characterized in that:
- in a first phase, a fully protected equivalent of the desired octasaccharide (I) is
synthesized,
- in a second phase, the positions that are to bear sulfonate groups on the final
molecule are deprotected and then O-sulfated,
- in a third phase, the whole compound is deprotected, and
- in a fourth phase, the N-acyl groups are introduced (introduction of groups the
R3-CO-).
The synthesis of the fully protected equivalent of the desired octasaccharide (I)
is performed according to reactions that are well known to those skilled in the art, and
using methods for the synthesis of oligosaccharides (for example G.J. Boons,
Tetrahedron (1996), 52, 1095-1121 and patent applications WO 98/03554 and
WO 99/36443), in which a glycoside bond-donating oligosaccharide is coupled with a
glycoside bond-accepting oligosaccharide to give another oligosaccharide whose size is
equal to the sum of the sizes of the two reactive species. This sequence is repeated
until the compound of formula (I) is obtained, optionally in protected form. The nature
and profile of the charge of the final desired compound determine the nature of the
chemical species used in the various synthetic steps, according to the rules well known
to those skilled in the art. Reference may be made, for example, to C. Van Boeckel and
M. Petitou, Angew. Chem. Int. Ed. Engl. (1993), 32, 1671-1690 or alternatively to H.
Paulsen, "Advances in selective chemical syntheses of complex oligosaccharides",
Angew. Chem. Int. Ed. Engl. (1982), 21, 155-173.
The compounds of the invention may naturally be prepared using various
strategies known to those skilled in the art of oligosaccharide synthesis. The process
described above is the preferred process of the invention. However, the compounds of
formula (l)/(l') may be prepared via other well-known methods of sugar chemistry,
described, for example, in "Monosaccharides, their chemistry and their roles in natural
products", P.M. Collins and R.J. Ferrier, J. Wiley & Sons (1995) and by G.J. Boons in
Tetrahedron (1996), 52, 1095-1121.
The protecting groups used in the process for preparing the compounds of
formula (l)/(l') are those that make it possible firstly to protect a reactive function such
as a hydroxyl or an amine during a synthesis, and secondly to regenerate the intact
reactive function at the end of the synthesis. The protecting groups commonly used in
sugar chemistry, as described, for example, in "Protective Groups in Organic
Synthesis", Greene et al., 3rd edition (John Wiley & Sons, Inc., New York) are used to
perform the process according to the invention. The protecting groups are chosen, for
example, from acetyl, methyl, pentenyl, benzoyl, levulinyl, benzyl, substituted benzyl,
azide, benzyl carbamate, te/f-butyldimethylsilyl (tBDMS) and ferf-butyldiphenylsilyl
(tBDPS) groups.
Activating groups may also be used; these are the groups conventionally used in
sugar chemistry, for example according to G.J. Boons, Tetrahedron (1996), 52, 1095-
1121. These activating groups are chosen, for example, from imidates and
thioglycosides.
The process described above allows the compounds of the invention to be
obtained in the form of salts, advantageously in the form of the sodium salt. To obtain
the corresponding acids, the compounds of the invention in salt form may be placed in
contact with a cation-exchange resin in acidic form. The compounds of the invention in
acid form may then be neutralized, with a base to obtain the desired salt. For the
preparation of the salts of the compounds of formula (l)/(l'), any mineral or organic base
that gives pharmaceutically acceptable salts with the compounds of formula (l)/(l') may
be used.
A subject of the invention is also the compounds of formula (V) below, in which
Alk represents an alkyl group, R1 is as defined previously in relation with the compounds
of formula (l)/(l'), and Pg, Pg' and Pg", which may be identical or different, represent
protecting groups:

Such compounds are useful as intermediates in the synthesis of the compounds
of formula (l)/(l').
In particular, a subject of the invention is the compounds (V) in which Alk
represents a methyl group and Pg, Pg' and Pg" represent, respectively, benzyl, tBDPS
and acetyl groups. More particularly, among these compounds of formula (V), the
invention is directed towards those in which R1, is chosen from the following groups: -O-
methyl, -O-(CH2)5-phenyl, -O-CH2-CH(C3H7)2, -O-(CH2)3-C6H11 and -O-(CH2)2-
CH(phenyl)2.
Such compounds correspond to disaccharides 58, 131, 132, 133 and 134
illustrated in the synthetic schemes below, which are useful for the synthesis of
compounds 10, 11 and 14 to 19 according to the invention, as will be detailed
hereinbelow.
A subject of the invention is also the compounds of formula (VI) below, in which
Alk represents an alkyl group and Pg, Pg' and Pg", which may be identical or different,
represent protecting groups:

Such compounds are useful as intermediates in the synthesis of the compounds
of formula (l)/(l'),
In particular, a subject of the invention is compound (VI) in which Alk represents
a methyl group and Pg, Pg' and Pg" represent, respectively, benzyl, levulinyl and acetyl
groups. Such a compound corresponds to disaccharide 24 illustrated in the synthetic
schemes below, which is useful for the synthesis of compounds 1 to 4 according to the
invention, as will be detailed hereinbelow.
The examples that follow describe the preparation of certain compounds in
accordance with the invention. These compounds are not limiting, but serve merely to
illustrate the present invention. The starting compounds and the reagents, when their
mode of preparation is not expressly described, are commercially available or described
in the literature, or else may be prepared according to methods that are described
therein or that are known to those skilled in the art.
The following abbreviations are used:
[a]D: optical rotation
Ac: acetyl
Bn: benzyl
TLC: thin-layer chromatography
DDQ: 2,3-dichloro-5,6-dicyano-1,4-benzoquinone
ESI: Electron-Spray Ionization
h: hours
LC-MS (*): liquid chromatography coupled to mass spectrometry
Lev: levulinyl
min: minutes
Me: methyl
PMB: para-methoxybenzyl
Rf: Retardation factor (retention time measured on TLC relative to the solvent
migration front)
tBDMS: terf-butyldimethylsilyl
tBDPS: tert-butyldiphenylsilyl
TFA: trifluoroacetic acid
TR: retention time measured by LC-MS*
Z: benzyloxycarbonyl
*The LC-MS are acquired on a Waters brand ZQ4000 machine. The column
used is a Symmetry C18 3.5 urn (2.1x50 mm). Eluent A is formed from H2O + 0.005%
TFA, pH 3.15. Eluent B is formed from acetonitrile + 0.005% TFA. The gradient ranges
from 0 to 90% of eluent B over 10 (or 30) minutes + 5 minutes at 90% of eluent B. The
flow rate is 0.4 mL/min.
Preparation of (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-idopyranosvl-
uronate)-(1?4)-2-amino-1.6-anhvdro-3-O-benzyl-2-deoxy-ß--D-qlucopvranose (No. 21)
To a solution of compound 20 (21.6 g, 31 mmol) (C.A.A. Van Boeckel et al., J.
Carbohydr. Chem., 4 (1985) 293-321) in N,N-dimethylformamide (155 mL) is added
triethylamine (Et3N) (65 mL, 15 molar equivalents), followed by addition of 1,3-
propanedithiol (47 mL, 15 moiar equivalents). After 17 hours of magnetic stirring, the
reaction mixture is diluted with ethyl acetate, washed with water, dried (Na2SO4), filtered
and concentrated, and then purified on silica gel (acetone - cyclohexane) to give 21
(15.8 g, 76%).
LC-MS m/z 672.2 [(M + H)+]. TR = 0.94 min
Preparation of (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-idopvranosvl-
uronate)-(1?4)-1,6-anhvdro-3-O-benzyl-2-deoxy-2-[(benzyloxy)carbonynamino-ß--D-
glucopyranose (No. 22)
To a solution of compound 21 (128.12 g, 190.7 mmol) in dichloromethane
(240 mL) is added saturated aqueous sodium hydrogen carbonate solution (240 mL),
followed by addition, at 0°C, of benzyloxycarbonyl chloride (35.5 mL, 1.3 molar
equivalents). After 15 minutes of vigorous magnetic stirring at 0°C, the reaction mixture
is diluted with dichloromethane, washed with water to neutrality, dried (Na2SO4), filtered
and concentrated. The crude product is used in the following step without purification.
LC-MS m/z 806.2 [(M + H)+]. TR = 1.55 min
Preparation of (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-idopvranosyl-
uronate)-(1?4)-1,6-di-O-acetyl-3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonyllamino-a,ß-
D-glucopvranose (No. 23)
The residue obtained in the preceding step is dissolved in acetic anhydride
(1.05L), followed by addition, at 0°C and over 15 min, of trifluoroacetic acid (TFA)
(105 mL). The reaction mixture is stirred for 10 minutes at 0°C and for 18 hours at room
temperature, and is then concentrated, co-evaporated with toluene, and purified on
silica gel (cyclohexane - acetone), to give compound 23 (138.9 g, 80%, 2 steps).
LC-MS m/z 930.3 [(M + Na)+]. TR = 21.68/21.84 min
Preparation of (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-idopvranosvl-
uronate)-(1?4)-6-O-acetyl-3-O-benzyl-2-deoxy-2-f(benzyloxy)carbonvnamino-a,ß--D-
glucopyranosetrichloroacetimidate (No. 24)
To a solution of compound 23 (25 g, 27.5 mmol) in diethyl ether (1.1 L) is added,
at 0°C, benzylamine (BnNH2) (109 mL, 36 molar equivalents). After stirring for 1 hour at
0°C, and then for 4.5 hours at room temperature, the reaction mixture is diluted with
ethyl acetate, and then neutralized with cold 1N HCI (0-4°C), washed with water, dried
(Na2SO4), filtered and concentrated, and purified on silica gel (acetone - cyclohexane) to
give (methyl 2-O-acetyl-3-O-benzyl-4-O-levulinoyl-a-L-idopyranosyluronate)-(1?4)-6-O-
acetyl-3-O-benzyl-2-deoxy-2-[(benzyloxy)carbonyl]amino-a,|3--D-glucopyranose (18.5 g,
78%).
LC-MS m/z 888.3 [(M + Naf]. TR = 19.95/20.60 min
To a solution of the compound thus obtained (18.5 g, 21.4 mmol) in
dichloromethane (405 mL) is added, under argon and at 0°C, caesium carbonate
(Cs2C03) (11.1 g, 1.6 molar equivalents) and trichloroacetonitrile (CCI3CN) (14.8 mL,
5.0 molar equivalents). After stirring for 1 hour at room temperature, the reaction
mixture is filtered and then concentrated. The residue is purified on silica gel (acetone -
cyclohexane + 0.1% triethylamine) to give 24 (17.2 g, 80%).
Rf = 0.47 (7/3 ethyl acetate/cyclohexane).
Chemical shifts of anomeric protons (500 MHz, CDCI3) 6 5.17 IdoUA", 6.28 Glc'a
LC-MS m/z 866.3 [(M + Na)+]. TR1 = 19.91 min
Preparation of 4-pentenyl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-
idopvranosvluronate)-(1?4)-6-O-acetyl-3-O-benzyl-2-deoxy-2-f(benzyloxy)carbonvn-
amino-B--D-qlucopyranoside (No. 25)
A solution of tert-butyldimethylsilyl triflate in dichloromethane (1M, 0.2 mol per
mole of imidate) is added, under argon and at -20°C, to a solution of the imidate 24
(13.7 g, 13.56 mmol) and of allyl alcohol (6.8 mL, 5 molar equivalents) in
dichloromethane (590 mL) in the presence of 4 A molecular sieves (17.6 g). After
20 minutes of stirring (TLC), saturated aqueous sodium hydrogen carbonate solution is
added. After filtering, washing with water, drying (Na2SO4), filtering and evaporating to
dryness, the mixture is purified on silica gel to give compound 25 (10.7 g, 85%).
LC-MS m/z 934.5 [(M + H)+]. TR = 24.00 min
Preparation of 4-pentenvl (methyl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvl-
uronate)-(1?4)-6-O-acetyl-3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonvnamino-3--D-
glucopyranoside (No. 26)
To a solution of compound 25 (10.6 g, 11.35mmol) in a 1:2 mixture of
toluene/ethanol (1.2 L) is added hydrazine acetate (5.2 g, 5 molar equivalents). After
2.5 hours of magnetic stirring, the mixture is concentrated under vacuum and then
purified on silica gel to give compound 26 (6.98 g, 74%).
LC-MS m/z 858.4 [(M + Na)+]. TR = 22.62 min
Preparation of 4-pentenvl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L_-
idopvranosvluronate)-(1?4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-gluco-
pyranosvP-(1?4)-(methvl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-(1?4)-6-O-
acetyl-3-O-benzyl-2-deoxy-2-[(benzyloxy)carbonyl]amino-3--D-qlucopyranoside (No. 28)
A solution of terf-butyldimethylsilyl triflate in dichloromethane (1M, 0.15 mol per
mole of imidate) is added, under argon and at -20°C, to a solution of the imidate 27
(9.0 g, 9.98 mmol) (C. Tabeur et a!., BioOrg. Med. Chem. (1999) 7, 2003-2012) and of
the glycosyl acceptor 26 (6.95 g, 8.32 mmol) in dichloromethane (350 mL) in the
presence of 4 A molecular sieves (7.5 g). After 18 hours at -20°C (TLC), solid sodium
hydrogen carbonate is added. After filtering, washing with aqueous 2% sodium
hydrogen carbonate solution, with water, drying (Na2SO4), filtering and evaporating to
dryness, the residue is purified on silica gel to give 28 (9.04 g, 69%).
Rf = 0.38 (1/4 acetone/toluene).
Preparation of 4-pentenvl (methyl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvl-
uronate)-(1?4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-alucopvranosvl)-(1?4)-
(methvl 2-O-acetyl-3-O-benzyl-a-L-idopvranosvluronate)-(1?4)-6-O-acetyl-3-O-benzyl-
2-deoxy-2-r(benzyloxy)carbonvnamino-3--D-qlucopyranoside (No. 29)
To a solution of compound 28 (9.0 g, 5.7 mmol) in a 1:2 mixture of
toluene/ethanol (570 mL) is added hydrazine acetate (2.6 g, 5 molar equivalents). After
1 hour of magnetic stirring, the mixture is concentrated under vacuum and then purified
on silica gel to give compound 29 (7.15 g, 85%).
Rf = 0.29 (1/4 acetone/toluene).
Preparation of 4-pentenvl (methyl 2-O-acetyl-3-O-benzyl-4-O-levulinoyl-a-L-
idopvranosvluronate)-(1?4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-gluco-
pvranosvlMI -»4)-(methvl 2-O-acetyl-3-O-benzyl-a-L-idopyranosyluronate)-(1 -+4H6-O-
acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-alucopvranosvlV(1?4)-(methvl 2-O-acetyl-3-O-
benzyl-a-L-idopvranosvluronate)-(1?4)-6-O-acetyl-3-O-benzyl-2-deoxy-2-r(benzyl-
oxy)c'arbonvnamino-B--D-qlucopyranoside (No. 30)
A solution of ferf-butyldimethylsilyl triflate in dichloromethane (1M, 0.15 mol per
mole of imidate) is added, under argon and at -20°C, to a solution of the imidate 27
(4.80 g, 5.32 mmol) and of the glycosyl acceptor 29 (7.15 g, 4.84 mmol) in
dichloromethane (180 mL) in the presence of 4 A molecular sieves (3.99 g). After
18 hours at -20°C (TLC), solid sodium hydrogen carbonate is added. After magnetic
stirring at room temperature (30 minutes), filtering, washing with aqueous 2% sodium
hydrogen carbonate solution, and then with water, drying (Na2SO4) and evaporating to
dryness, the residue is purified on silica gel to give 30 (5.23 g, 49%).
Rf = 0.18 (1/4 acetone/diisopropyl ether).
Preparation of 4-pentenvl (methyl 2-O-acetyl-3-O-benzyl-a-L-idopvranosyl-
uronate)-(1?4))-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosyln-(1?4))-
(methyl 2-O-acetyl-3-O-benzyl-a-L-idopvranosvluronate)-(1?4)-(6-O-acetyl-2-azido-3-
0-benzyl-2-deoxy-a-D-qlucopyranosyll)-(1?4)-(methvl______2-O-acetyl-3-O-benzyl-a-L-
idopvranosvluronate)-(1?4)-6-O-acetyl-3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonvn-
amino-B--D-glucopyranoside (No. 31)
To a solution of compound 30 (5.21 g, 2.35 mmol) in a 1:2 mixture of
toluene/ethanol (180 mL) is added hydrazine acetate (1.08 g, 5 molar equivalents). After
1 hour 10 minutes of magnetic stirring, the mixture is concentrated under vacuum and
then purified on silica gel to give compound 31 (4.78 g, 96%).
Rf = 0.47 (1/3 acetone/diisopropyl ether).

Preparation of (3,4-di-O-benzyl-2-O-(4-methoxy)benzyl-6-O-fe/f-butvldimethyl-
silvl-a-L-idopvranosvl)-(1^-4)-1,6-anhvdro-2-azido-3-O-benzyl-2-deoxy-3--D-qluco-
pyranose (No. 33)
To a solution of compound 32 (32.3 g, 42.2 mmol) (WO 2006/021 653) in N,N-
dimethylformamide (210 ml_) is added, at 0°C and under argon, benzyl bromide (25 mL»
5 molar equivalents), followed by addition of 55% NaH (3 g, 1.5 molar equivalents).
After 20 minutes of magnetic stirring, methanol is added (30 mL), the reaction medium
is concentrated under vacuum, and the crude reaction product is diluted with ethyl
acetate, washed with water and then with saturated aqueous sodium chloride solution,
dried (Na2SO4), filtered and concentrated. The residue obtained is used in the following
step without purification.
LC-MS m/z 871.7 [(M + NH4)+]. TR = 13.86 min
Preparation of (2-O-acetyl-3,4-di-O-benzyl-6-O-tert-butvldimethvlsilvl-a-L-
idopvranosvl)-(1?4))-1,6-anhvdro-2-azido-3-O-benzyl-2-deoxy-3--D-glucopyranose (No.
34)
To a solution of crude 33 (38.6 g) in dichloromethane (1.6 L) is added water
(80 mL), followed by addition, at 0°C, of DDQ (14.2 g, 1.5 molar equivalents). After
4 hours 45 minutes of stirring at 0°C, the medium is diluted with dichloromethane and
sodium hydrogen carbonate solution is added. The organic phase is then washed with
water, dried (Na2SO4), filtered and concentrated. The compound obtained is used in the
following step without purification.
The residue obtained is dissolved in dichloromethane (350 mL), followed by
addition of triethylamine (13 mL), 4-dimethylaminopyridine (2 g) and acetic anhydride
(60 mL). After magnetic stirring for 10 minutes at 0°C and then for 1 hour 45 minutes at
room temperature, the reaction mixture is diluted with dichloromethane and then
washed successively with aqueous 10% potassium hydrogen sulfate solution and water,
and the organic phase is then dried (Na2SO4), filtered and concentrated.
The residue obtained is purified on silica (ethyl acetate - cyclohexane) to give 34
(26.8 g, 83%, 3 steps).
LC-MS m/z 798.3 [(M + Na)+]. TR = 12.97 min
Preparation of (methyl 2-O-acetyl-3,4-di-O-benzyl-a-L-idopvranosvluronate)-
(1—»4)-1,6-anhvdro-2-azido-3-O-benzyl-2-deoxy-B--D-qlucopvranose (No. 35)
To a solution of 34 (26.3 g, 33.9 mmol) in acetone (1.4 L) is added, at 0°C, a
solution of Cr03 (10.5 g, 3.1 molar equivalents) in aqueous 3.5 M H2SO4 (47 mL), After
mechanical stirring for 4 hours at 0°C, the reaction medium is diluted with
dichloromethane, washed with water to neutrality, and the organic phase is then dried
(Na2SO4), filtered and concentrated. The compound obtained is used in the following
step without purification.
The residue obtained is dissolved in A/,A/-dimethylformamide (210 ml_), and
potassium hydrogen carbonate (17 g, 5 molar equivalents) and methyl iodide (21 mL,
10 molar equivalents) are added. The reaction mixture is stirred at room temperature for
16 hours, and then concentrated under vacuum. The residue is diluted with ethyl
acetate and then washed with water, with saturated aqueous sodium thiosulfate
solution, with saturated aqueous sodium chloride solution, and then dried (Na2SO4),
filtered and concentrated. The compound obtained is used in the following step without
purification.
LC-MS m/z 707.3 [(M + NH4)+]. TR = 10.37 min
Preparation of (methyl 2-O-acetyl-3,4-di-O-benzyl--D-i_-idopvranosvluronate)-
(1?4)-1,6-di-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a.B--D-qlucopyranose (No. 36)
The crude residue obtained in the preceding step is dissolved in acetic anhydride
(177 mL), followed by addition of trifluoroacetic acid (TFA) (17.7 mL). The reaction
mixture is stirred for 16 hours, and is then concentrated, co-evaporated with toluene,
and purified on silica gel (cyclohexane - ethyl acetate), to give compound 36 (17.4 g,
65%, 3 steps).
LC-MS m/z 809.3 [(M + NH4)+]. TR = 10.81 min
Preparation of (methyl 2-O-acetyl-3,4-di-O-benzyl-a-L-idopvranosvluronate)-
(1?4)-6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a.B--D-alucopyranose (No. 37)
To a solution of compound 36 (7 g, 8.84 mmol) in diethyl ether (303 mL) is
added, at 0°C and under argon, benzylamine (BnNH2) (29.7 mL, 36 molar equivalents).
After 1 hour of magnetic stirring at 0°C and then 6 hours at room temperature, the
reaction mixture is neutralized with cold (0-4°C) 1N HCI, washed with water, dried
(Na2SO4), filtered and concentrated, and purified on silica gel (ethyl acetate -
cyclohexane) to give 37 (5.95 g, 90%).
LC-MS m/z 767.7 [(M + NH4)+]. TR = 1.64 min
Preparation of (methyl 2-O-acetyl-3,4-di-O-benzyl--D-i_-idopvranosvluronateV
(1?4)-6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a,B--D-qlucopyranose trichloroacetimidate
(No. 38)
To a solution of compound 37 (5.94 g, 7.9 mmol) in dichloromethane (150 mL) is
added, under argon, caesium carbonate (Cs2C03) (4.1 g, 1.6 molar equivalents),
followed by addition of trichloroacetonitrile (CCI3CN) (3.9 mL, 5.0 molar equivalents).
After stirring for 45 minutes at room temperature, the reaction mixture is filtered and
then concentrated. The residue is purified on silica gel (ethyl acetate - cyclohexane +
0.1% triethylamine) to give 38 (5.7 g, 81%).
LC-MS m/z 912.0 [(M + NH4)+]. TR = 1.81 min

Preparation of 4-pentenvl (methyl 2-O-acetyl-3.4-di-O-benzyl-a-L-idopvranosvl-
uronate)-(1?4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosvU-(1?4)-
(methvl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-(1?4)-(6-O-acetyl-2-azido-3-
O-benzyl-2-deoxy-o--D-glucopvranosy0-(1—»4)-(methvl______2-O-acetyl-3-O-benzyl-g-i_-
idopvranosvluronate)-(1?4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-g--D-gluco-
pyranosylHI —>4)-(methvl 2-O-acetyl-3-O-benzyl-g-L-idopyranosvluronate)-(1?4)-6-O-
acetyl-3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonvnamino-B--D-glucopyranoside (No. 39)
A solution of terf-butyldimethylsilyl triflate in dichloromethane (1M, 0.1 mol per
mole of imidate) is added, under argon and at -15°C, to a solution of the imidate 38
(2.61 g, 2.92 mmol) and of the glycosyl acceptor 31 (4.78 g, 2.25 mmol) in
dichloromethane (146 mL) in the presence of 4 A molecular sieves (1.99 g). After
2 hours of magnetic stirring at -15°C (TLC), imidate 38 is again added at time intervals
of between 15 and 30 minutes, to maximum consumption of the acceptor 31. At the end
of the reaction (TLC), solid sodium hydrogen carbonate is added to neutralize, and after
magnetic stirring at room temperature (15 minutes), filtering, washing with aqueous 2%
sodium hydrogen carbonate solution, and then with water, drying (Na2SO4) and
evaporating to dryness, the residue is purified on silica gel to give 39 (3.71 g, 58%).
Rf = 0.21 (15/85 acetone/toluene).
Preparation of 4-pentenyl (methyl 3,4-di-O-benzyl-a-L-idopyranosvluronate)-
n^4H2-azido-3-O-benzyl-2-deoxy-O--D-qlucopyranosvn-(1?4Hmethvl 3-O-benzyl-a-L-
idopvranosvluronateV(1^4V(2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosvl)-(1?4))-
(methyl 3-O-benzyl-a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-a-D-
qlucopyranosvlH1?4)-(methyl 3-O-benzyl-O-L-idopvranosvluronateM1?4V3-O-benzyl-
2-deoxy-2-f(benzyloxy)carbonvnamino-3--D-qlucopyranoside (No. 40)
To a solution of 39 (300 mg, 0.105 mmol) in a 3/2 mixture of
methanol/dichloromethane (31 ml_) is added, at 0°C, a 1M a methanolic solution of
sodium methoxide (0.31 mL), in the presence of 3 A molecular sieves. After stirring at
0°C for 1 hour and at room temperature for 15 hours, the mixture is neutralized with H+
Dowex 50WX4 resin. After filtering and partially concentrating, the reaction mixture is
purified on an LH-20 exclusion column using a 1:1 methanol/dichloromethane mixture
as eluent, and, after pooling the fractions containing the product and concentrating
under vacuum, the residue is purified on silica gel with a dichloromethane/ethyl
acetate/ethanol mixture to give compound 40 (185 mg, 69%).
Rf = 0.22 (9/1/0.3 CH2CI2/EtOAc/EtOH).
Preparation of 4-pentenvl (methyl 3.4-di-O-benzyl-2-O-triethvlammonium
sulfonato-a-L-idopvranosvluronateV(1?4))-(2-azido-3-O-benzyl-2-deoxy-6-O-triethvl-
ammonium sulfonato-a-D-qlucopyranosyllV( 1 -^-(methyl 3-O-benzyl-2-O-triethvl-
ammonium sulfonato-a-L-idopvranosyluronate)-( 1?4 H2-azido-3-O-benzyl-2-deoxy-6-
O-triethvlammonium sulfonato-a-D-glucopvranosvlH1?4Mmethvl 3-O-benzyl-2-O-
triethylammonium sulfonato-g-L-idopvranosvluronateM1?4)-(2-azido-3-O-benzyl-2-
deoxy-6-O-triethylammonium sulfonato-g--D-glucopvranosvlH1-*4)-(methvl 3-O-benzyl-
2-O-triethylammonium sulfonato-O-i_-idopvranosvluronateH1?4)-3-O-benzyl-2-deoxy-2-
[(benzyloxy)carbonvnamino-6-O-triethvlammonium sulfonato-B--D-glucopyranoside (No.
41)
Triethylamine/sulfur trioxide complex (513 mg, 5 mol per hydroxyl function) is
added to a solution in A/,A/-dimethylformamide (6.3 ml_, 90 L/mol) of compound 40
(178 mg, 70.8 umol). After 17 hours of magnetic stirring at 55°C sheltered from light,
methanol is added at 0°C and, after stirring for 30 minutes at 0°C and then for
30 minutes at room temperature, the reaction medium is purified by means of an LH-20
column, using a 9:1 mixture of methanol/N,/V-dimethylformamide as eluent. The
fractions containing the product are then concentrated under high vacuum to give the
desired compound 41 (241 mg, 87%).
Rf = 0.33 (11/7/1.6/4 EtOAc/pyridine/AcOH/H20).
Preparation of 4-pentenvl (lithium 3,4-di-O-benzyl-2-O-lithium sulfonato-g-L-
idopvranosvluronateH1-+4M2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-g--D-
glucopyranosvlH1?4Hlithium 3-O-benzyl-2-O-lithium sulfonato-g-L-idopyranosvl-
uronateV(1^4V(2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-g--D-glucopyranosvD-
(1?4Hlithium 3-O-benzyl-2-O-lithium sulfonato-g-L-idopyranosvluronateH1?4H2-
azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-g--D-glucopyranosvlM1?4Hlithium 3-
O-benzyl-2-O-lithium sulfonato-g-L-idopyranosvluronate)-(1-»4')-3-O-benzyl-2-deoxy"2-
r(benzyloxy)carbonvnamino-6-O-lithiumsulfonato-B--D-glucopyranoside (No. 42)
To a solution of compound 41 (238 mg, 60 umol) in a 1/1 mixture of
methanol/tetrahydrofuran (9.6 mL) is added, at 0°C, a molar aqueous LiOH solution.
After stirring for 1 hour at 0°C and then for 16 hours at room temperature, the reaction
medium is purified by means of an LH-20 column, using a 9:1 mixture of methanol/N,N-
dimethylformamide as eluent. The fractions containing the product are then
concentrated under high vacuum to give the desired compound 42.
Rf = 0.42 (11/7/1.6/4 EtOAc/pyridine/AcOH/H20).
Preparation of pentvl (sodium 2-O-sodium sulfonato-a-L-idopyranosvluronateV
(1?4H2-amino-2-deoxy-6-O-sodium sulfonato-a-D-qlucopyranosylD-(1?4Msodium 2-
O-sodium sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-amino-2-deoxy-6-O-sodium
sulfonato-a-D-qlucopyranosvlM1?4Hsodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronateHI —>4M2-amino-2-deoxy-6-O-sodium sulfonato-a-D-glucopvranosvl)-(1?4)-
(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronateV1?4)-2-amino-2-deoxy-6-O-
sodium sulfonato-B--D-glucopyranoside (No. 43)
To a solution in a 1/1 mixture of te/t-butanol/water (14 ml_) of compound 42
obtained previously are successively added ammonium formate (617 mg, 9.8 mmol)
and then 10% Pd/C (1.55 g). After stirring vigorously for 4 hours at room temperature,
the reaction medium is filtered and partially concentrated under vacuum, and the
solution is then applied to a Sephadex® G-25 column eluted with 0.2 M sodium chloride.
The fractions containing the product are concentrated and desalified using the same
column eluted with water. The fractions containing the product are then concentrated
under high vacuum to give the desired compound 43 (123 mg, 61% (2 steps)).
Chemical shifts of the anomeric protons (600 MHz, D20) 5 5.11 ldoUAvl", 5.38
Glcv", 5.19 ldoUAvl, 5.38 Glcv, 5.19 ldoUAlv, 5.38 Glc'", 5.21 IdoUA", 4.70 Glc'
Preparation of (Z/E) undec-4-enyl (methyl 2-O-acetyl-3.4-di-O-benzyl-a-L-
idopvranosvluronate)-(1^-4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qluco-
pyranosvlMI —>4)-(methvl 2-O-acetyl-3-O-benzyl-a-L-idopvranosvluronateM1?4)-(6-O-
acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-glucopyranosvl)-(1—»4Mmethvl 2-O-acetyl-3-O-
benzyl-a-L-idopvranosvluronate)-(1?4))-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-
qlucopyranosylD-d —>4)-(methvl 2-O-acetyl-3-O-benzyl-o-L-idopvranosyluronate)-(1?4)-
6-O-acetyl-3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonvnamino-B--D-alucopvranoside (No.
44)
To a solution of compound 39 (610 mg, 214 umol) in dichloromethane (10.7 ml_)
are added 1-octene (67 uL, 2 molar equivalents) and first-generation Grubbs catalyst
(C44H74CI2P2RU) (18 mg, 0.1 molar equivalent). After refluxing for 6 hours and then
stirring overnight at room temperature, the mixture is concentrated under high vacuum
and may be used under the same conditions in the case of an incomplete reaction. The
crude reaction product is purified by chromatography on silica gel (toluene-acetone) to
give the desired product 44 (131 mg, 20%).
Rf = 0.18 (85/15 toluene/acetone).
Preparation of (ZJE) 5-phenvl-4-pentenvl (methyl 2-O-acetyl-3,4-di-O-benzyl-g-L-
idopvranosyluronate)-(1?4H6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qluco-
pyranosvP-d —>4)-(methyl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronateM1?4)-(6-O-
acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosvD-(1?4)-(methyl 2-O-acetyl-3-O-
benzyl-a-L-idopvranosyluronate)-(1?4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-
qlucopyranosvlM1?4)-(methvl 2-O-acetyl-3-O-benzyl-O-i_-idopvranosvluronateH1?4)-
6-O-acetyl-3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonyllamino-3--D-qlucopyranoside (No.
45)
To a solution of compound 39 (100 mg, 35 umol) in dichloromethane (1 mL) are
added styrene (16 uL, 4 molar equivalents) and second-generation Grubbs catalyst
(C46H65CI2N2PRu) (1.8 mg, 0.06 molar equivalent). After refluxing for 8 hours and then
stirring overnight at 35°C, the reagents are optionally added, if necessary. The mixture
is then concentrated under high vacuum and then purified by chromatography on silica
gel (toluene-ethyl acetate) to give the desired product 45 (38 mg, 37%).
Rf = 0.25 (toluene-EtOAc 7/3).
Preparation of (ZJE) undec-4-envl (methyl 3.4-di-O-benzyl-a-L-idopyranosvl-
uronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosyll)-(1?4)-(methvl 3-O-
benzyl-a-L-idopvranosyluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-a-D-qluco-
pvranosvl)-(1?4)-(methyl 3-O-benzyl-a-L-idopyranosvluronate)-(1—»4)-(2-azido-3-O-
benzyl-2-deoxy-a-D-glucopvranosvlH1?4)-(methyl_______3-O-benzyl-O-L-idopvranosyl-
uronate)-(1?4)-3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonvnamino-3--D-qlucopvranoside
(No. 46)
To a solution of 44 (152 mg, 0.052 mmol) in a 3/2 mixture of
methanol/dichloromethane (15 mL) is added, at 0°C, a 1M a methanolic solution of
sodium methoxide (154 uL), in the presence of 3 A molecular sieves. After stirring at
0°C for 1 hour and at room temperature for 15 hours, the mixture is neutralized with H+
Dowex 50WX4 resin. After filtering and partially concentrating, the reaction mixture is
purified on an LH-20 exclusion column using a 1/1 mixture of methanol/dichloromethane
as eluent, and, after pooling the fractions containing the product and concentrating
under vacuum, the residue is purified on silica gel with a toluene/acetone/ethyl
acetate/ethanol mixture to give compound 46 (83 mg, 61%).
Rf = 0.25 (7/3/0.1 toluene/acetone/EtOH).
Preparation of (ZIE) 5-phenvl-4-pentenvl (methyl 3.4-di-O-benzyl-a-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosvlH1?4)-
(methvl 3-O-benzyl-a-L-idopyranosvluronate)-(1-»-4)-(2-azido-3-O-benzyl-2-deoxy-a-D-
glucopyranosvD-d -»4)-(methvl 3-O-benzyl-a-L-idopvranosvluronateH1?4)-(2-azido-3-
0-benzyl-2-deoxy-a-D-qlucopyranosyllH1?4)-(methvl 3-O-benzyl-a-L-idopvranosvl-
uronate)-(1?4)-3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonyllamino-B--D-qlucopyranoside
(No. 47)
To a solution of 45 (229 mg, 0.078 mmol) in a 3/2 mixture of
methanol/dichloromethane (23 mL) is added, at 0°C, a 1M a methanolic solution of
sodium methoxide (232 uL), in the presence of 3 A molecular sieves. After stirring at
0°C for 1 hour and at room temperature for 15 hours, the mixture is neutralized with H+
Dowex 50WX4 resin. After filtering and partially concentrating, the reaction mixture is
purified on an LH-20 exclusion column using a 1/1 mixture of methanol/dichloromethane
as eluent, and, after pooling the fractions containing the product and concentrating
under vacuum, the residue is purified on silica gel with a dichloromethane/ethyl
acetate/ethanol mixture to give compound 47 (141 mg, 69%).
Rf = 0.15 (7/3/0.1 toluene/acetone/EtOH).
Preparation of (Z/E) undec-4-envl (methyl 3.4-di-O-benzyl-2-O-triethylammonium
sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-triethvl-
ammonium sulfonato-a-D-qlucopyranosyll)-(1?4)-(methyl 3-O-benzyl-2-O-triethyl-
ammonium sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-
O-triethylammonium sulfonato-a-D-glucopvranosvlH1?4)-(methvl 3-O-benzyl-2-O-
triethylammonium sulfonato--D-i_-idopvranosyluronate)-( 1?4)-(2-azido-3-O-benzyl-2-
deoxy-6-O-triethvlammonium sulfonato-a-D-qlucopyranosyllH1?4Mmethyl 3-O-benzyl-
2-O-triethvlammonium sulfonato-a-L-idopyranosvluronate)-(1?4)-3-O-benzyl-2-deoxy-2-
f(benzyloxy)carbonvnamino-6-O-triethvlammonium sulfonato-B--D-qlucopyranoside (No.
48)
Triethylamine/sulfur trioxide complex (225 mg, 5 mol per hydroxyl function) is
added to a solution in A/,A/-dimethylformamide (2.8 mL, 90 L/mol) of compound 46
(81 mg, 31 Mmol). After 16 hours of magnetic stirring at 506C sheltered from light,
methanol is added at 0°C (0.2 ml_) and after 35 minutes of stirring at 0°C and then
2.5 hours at room temperature, the reaction medium is purified by means of an LH-20
column, using a 1/1/0.1 mixture of methanol/dichloromethane/N,N-dimethylformamide
as eluent. The fractions containing the product are then concentrated under high
vacuum to give the desired compound 48 (122 mg, 99%).
Rf = 0.36 (11/7/1.6/4 EtOAc/pyridine/AcOH/H20).
Preparation of (ZIE) 5-phenvl-4-pentenvl (methvl 3.4-di-O-benzyl-2-O-triethvl-
ammonium sulfonato-a-L-idopvranosvluronateH1?4M2-azido-3-O-benzyl-2-deoxy-6-
O-triethvlammonium sulfonato-a-D-qlucopyranosyllH1?4Mmethyl 3-O-benzyl-2-O-
triethvlammonium sulfonato-a-L-idopvranosvluronateM 1?4H2-azido-3-O-benzyl-2-
deoxy-6-O-triethvlammonium sulfonato-a-D-qlucopyranosvD-d —>4Mmethyl 3-O-benzyl-
2-O-triethvlammonium sulfonato-a-L-idopyranosvluronate)-(1?4H2-azido-3-O-benzyl-
2-deoxy-6-O-triethvlammonium sulfonato-a-D-qlucopyranosyllH1?4Hmethvl 3-O-
benzyl-2-O-triethylammonium sulfonato--D-i_-idopvranosyluronateH1—»4)-3-O-benzyl-2-
deoxy-2-f(benzyloxy)carbonyllamino-6-O-triethylammonium_______sulfonato-B--D-qluco-
pyranoside (No. 49)
Triethylamine/sulfur trioxide complex (385 mg, 5 mol per hydroxyl function) is
added to a solution in N,W-dimethylformamide (4.7 mL, 90 L/mol) of compound 47
(138 mg, 53 umol). After 16 hours of magnetic stirring at 50°C sheltered from light,
methanol is added at 0°C (0.35 mL) and after 30 minutes of stirring at 0°C and then
1 hour at room temperature, the reaction medium is purified by means of an LH-20
column, using a 1/1/0.1 mixture of methanol/dichloromethane/A/,A/-dimethylformamide
as eluent. The fractions containing the product are then concentrated under high
vacuum to give the desired compound 49 (194 mg, 92%).
Rf = 0.35 (11/7/1.6/4 EtOAc/pyridine/AcOH/H20).
Preparation of (Z/E) undec-4-enyl (lithium 3,4-di-O-benzyl-2-O-lithium sulfonato-
a-L-idopyranosvluronate)-(1-»-4)-(2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-g--D-
glucopyranosvlHI -»4Hlithium 3-O-benzyl-2-O-lithium sulfonato-a-L-idopyranosvl-
uronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-a-D-qlucopyranosvl)-
(1?4)-(lithium 3-O-benzyl-2-O-lithium sulfonato-a-L-idopyranosvluronate)-(1-»4V(2-
azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-a-D-qlucopyranosyll)-(1 -^4)-(lithium 3-
O-benzyl-2-O-lithium sulfonato-a-L-idopyranosvluronate)-(1?4)-3-O-benzyl-2-deoxy-2-
r(benzyloxy)carbonyllamino-6-O-lithium sulfonato-B--D-qlucopyranoside (No. 50)
To a solution of compound 48 (119 mg, 29 umol) in a 1/1 mixture of
methanol/tetrahydrofuran (4.7 mL) is added, at 0°C, a molar aqueous LiOH solution
(1.18 mL). After stirring for 1 hour at 0°C and then for 19.5 hours at room temperature,
the reaction medium is purified by means of an LH-20 column, using a 4:1 mixture of
methanol//V,N-dimethylformamide as eluent. The fractions containing the product are
then concentrated under high vacuum to give the desired compound 50.
Rf = 0.40 (11/7/1.6/4 EtOAc/pyridine/AcOH/H20).
Preparation of (ZJE) 5-phenvl-4-pentenvl (lithium 3,4-di-O-benzyl-2-O-lithium
sulfonato-a-L-idopvranosvluronateV(1^4V(2-azido-3-O-benzyl-2-deoxy-6-O-lithium
sulfonato-g--D-glucopvranosvlH1?4V(lithium 3-O-benzyl-2-O-lithium sulfonato-a-L-
idopvranosvluronateH1^4H2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-a-D-
glucopvranosvD-1?4Hlithium 3-O-benzyl-2-O-lithium sulfonato-a-L-idopyranosvl-
uronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-a-D-glucopyranosvl)-
(1?4Mlithium 3-O-benzyl-2-O-lithium sulfonato-a-L-idopyranosvluronate)-(1?4)-3-O-
benzyl-2-deoxy-2-[(benzyloxy)carbonvnamino-6-O-lithium_________sulfonato-B--D-gluco-
pyranoside (No. 51)
To a solution of compound 49 (190 mg, 47 umol) in a 1/1 mixture of
methanol/tetrahydrofuran (7.4 mL) is added, at 0°C, a molar aqueous LiOH solution
(1.88 mL). After stirring for 1 hour at 0°C and then for 16 hours at room temperature,
the reaction medium is purified by means of an LH-20 column, using a 9:1 mixture of
methanol/N, W-dimethylformamide as eluent. The fractions containing the product are
then concentrated under high vacuum to give the desired compound 51.
Rf = 0.44 (11/7/1.6/4 EtOAc/pyridine/AcOH/H20).
Preparation of undecyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-
(1?4)-(2-amino-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosv0-(1?4)-(sodium 2-
O-sodium sulfonato-a-L-idopvranosvluronateH1-+4H2-amino-2-deoxy-6-O-sodium
sulfonato-a-D-glucopvranosvlH1?4Hsodium 2-O-sodium sulfonato-a-L-idopyranosvl-
uronate)-(1?4M2-amino-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosvn-(1?4)-
(sodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-( 1?4)-2-amino-2-deoxy-6-O-
sodium sulfonato-3--D-glucopyranoside (No. 52)
To a solution in a 1/1 mixture of ferf-butanol/water (6 mL) of compound 50
obtained previously are successively added ammonium formate (260 mg, 4.1 mmol)
and then 10% Pd/C (672 mg). After 4 hours 5 minutes of vigorous stirring at room
temperature, the reaction medium is filtered and partially concentrated under vacuum,
and the solution is then applied to a Sephadex® G-25 column eluted with 0.2 M sodium
chloride. The fractions containing the product are concentrated and desalified using the
same column eluted with water. The fractions containing the product are then
concentrated under high vacuum to give the desired compound 52 (34 mg, 49%
(2 steps)).
Mass: "ESI" method, negative mode: theoretical mass = 2337.81; experimental
mass: 2337.27 ± 0.06 a.m.u. [M-4Na+4H].
Preparation of 5-phenvlpentvl (sodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronateV1?4)-(2-amino-2-deoxy-6-O-sodium sulfonato-o--D-qlucopyranosyllH1?4V
(sodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-(1?4H2-amino-2-deoxy-6-O-
sodium sulfonato-g--D-glucopvranosvlH1?4Hsodium 2-O-sodium sulfonato-g-L-
idopyranosyluronateMI—»4)-(2-amino-2-deoxy-6-O-sodium_______su Ifonato-a-D-ql u co-
pyranosvlH1?4V(sodium 2-O-sodium sulfonato-o-L-idopvranosvluronate)-(1?4)-2-
amino-2-deoxy-6-O-sodium sulfonato-B--D-qlucopvranoside (No. 53)
To a solution in a 1/1 mixture of ferf-butanol/water (9.2 mL) of compound 51
obtained previously are successively added ammonium formate (407 mg, 6.47 mmol)
and then 10% Pd/C (1.05 g). After 4 hours 5 minutes of vigorous stirring at room
temperature, the reaction medium is filtered and partially concentrated under vacuum,
and the solution is then applied to a Sephadex® G-25 column eluted with 0.2 M sodium
chloride. The fractions containing the product are concentrated and desalified using the
same column eluted with water. The fractions containing the product are then
concentrated under high vacuum to give the desired compound 53 (79 mg, 72%
(2 steps)).
"ESI" method, negative mode: multicharged ion detected m/z 537.0713 [M-4H]4"
(acid form).

Preparation of methyl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-o-L-
idopvranosvluronate)-(1?4)V6-O-acetyl-3-O-benzyl-2-benzyloxycarbonylamino-2-deoxy-
g--D-qlucopyranoside (No. 55)
To a solution of compound 54 (23.5 g, 30 mmol; Carbohydrate Research (1987),
167, 67-75) in dichloromethane (600 mL) are added, at 0°C and under an inert
atmosphere, 4-dimethyIaminopyridine (733 mg, 6 mmol), 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide hydrochloride (11.5 g, 60 mmol) and levulinic acid (6.2 mL; 60 mmol).
After stirring for 16 hours at room temperature, the mixture is diluted with
dichloromethane (1.5 L). The organic phase is washed successively with aqueous 10%
potassium hydrogen sulfate solution, with water, with saturated sodium hydrogen
carbonate solution and then with water, dried over sodium sulfate, filtered and then
evaporated to dryness. The residue is purified by flash chromatography on a column of
silica gel (1/3 cyclohexane/ethyl acetate) to give compound 55 (22.6 g).
Rf = 0.37, silica gel, 1/3 cyclohexane/ethyl acetate
Preparation of methyl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-
idopvranosvluronate)-(1?4)-3-O-benzyl-2-benzyloxycarbonylamino-2-deoxy-a-D-qluco-
pyranoside (No. 56)
To a solution of compound 55 (20.2 g, 23 mmol) in a 1/1
tetrahydrofuran/methanol mixture (140 mL) is added, under an inert atmosphere,
[tBu2SnCI(OH)]2 (226 mg, 0.79 mmol) prepared according to A. Orita et al., Chem. Eur.
J. (2001) 7, 3321. The reaction medium is stirred for 38 hours at 35°C. After
concentrating, the residue (20.8 g) is used without purification in the following step.
Rf = 0.23, silica gel, 1/3 cyclohexane/ethyl acetate
Preparation of methyl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-
idopvranosvluronate)-(1?4)-3-O-benzyl-2-benzyloxycarbonylamino-6-O-tert-butvl-
diphenvlsilvl-2-deoxy-a-D-qlucopvranoside (No. 57)
To a solution of the crude compound 56 (23 mmol) in dichloromethane (190 mL)
are added, at 0°C and under an inert atmosphere, triethylamine (8 mL, 57.5 mmol), 4-
dimethylaminopyridine (1.4 g, 11.5 mmol) and terf-butyldiphenylsilyl chloride (12 mL,
46.0 mmol). The reaction medium is stirred for 16 hours at room temperature. The
reaction mixture is diluted with dichloromethane. The organic phase is washed
successively with saturated sodium chloride solution and then with water, dried over
sodium sulfate, filtered and then evaporated. The residue is purified by flash
chromatography on a column of silica gel (2/1 cyclohexane/ethyl acetate) to give
compound 57 (24.4 g).
Rf = 0.42, silica gel, 2/1 cyclohexane/ethyl acetate
Preparation of methyl (methyl 2-O-acetyl-3-O-benzyl-a-L-idopvranosvluronate)-
(1?4)-3-O-benzyl-2-benzyloxycarbonylamino-6-O-feAf-butvldiphenylsilvl-2-deoxy-a-D-
glucopyranoside (No. 58)
To a solution of compound 57 (22.6 g, 20.0 mmol) in a 1/2 toluene/ethanol
mixture (2.5 L) is added hydrazine acetate (9.21 g, 100.0 mmol). The reaction medium
is stirred for 30 minutes at room temperature. After concentrating, the residue is purified
by flash chromatography on a column of silica gel (2/1 cyclohexane/ethyl acetate) to
give 17.6 g of compound 58.
Rf = 0.40, silica gel, 2/1 cyclohexane/ethyl acetate.
Chemical shifts of anomeric protons (500 MHz, CDCI3) 5 5.19 IdoUA", 4.63 Glc'
LC-MS m/z 978.5 [(M + H)+]. TR = 12.29 min
Preparation of (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl--D-i_-idopyranosvl-
uronate)-(1?4)-1-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranose (No. 60)
To a solution of 59 (11 g, 13.7 mmol) (C. Tabeur, et a/., Carbohydr. Res., 281
(1996) 253-276) in a 1/1 mixture of methanol/tetrahydrofuran (80 mL) is added
[tBu2SnCI(OH)]2 (0.55 g, 0.14 molar equivalent) prepared according to A. Orita et al.,
Chem. Eur. J. (2001) 7, 3321. After stirring at 35°C for 5.5 hours, then at room
temperature for 16 hours and then again at 35°C for 4 hours, the reaction mixture is
concentrated under vacuum and then purified by chromatography to give compound 60
(5.97 g, 57%).
LC-MS m/z 780.2 [(M + Na)+]. TR = 9.14 min
Preparation of (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-idopyranosvl-
uronate)-(1?4)-1-O-acetyl-2-azido-3-O-benzyl-6-O-te/t-butvldiphenvlsilvl-2-deoxy-a-D-
qlucopyranose (No. 61)
Compound 60 (5.97 g, 7.88 mmol) is dissolved in dichloromethane (63 mL). 4-
Dimethylaminopyridine (0.481 g, 0.5 molar equivalent), triethylamine (2.7 mL, 2.5 molar
equivalents), and terf-butyldiphenylsilyl chloride (4 ml_, 2 molar equivalents) are
successively added at 0°C under argon. After magnetic stirring for 4 hours, the reaction
medium is diluted with dichloromethane, washed with aqueous 10% potassium
hydrogen sulfate solution, with water, dried (Na2SO4), filtered and concentrated. The
residue obtained is purified on silica (ethyl acetate - heptane) to give 61 (7 g, 90%).
LC-MS m/z 1018.3 [(M + Na)+]. TR = 12.33 min
Preparation of (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-idopyranosvl-
uronate)-(1?4))-2-azido-3-O-benzyl-6-O-fe/t-butvldiphenvlsilyl-2-deoxy-a,B--D-gluco-
pyranose (No. 62)
To a solution of compound 61 (7 g, 7.03 mmol) in diethyl ether (70 mL) is added,
at 0°C, benzylamine (BnNH2) (29 mL, 38 molar equivalents). After 15 minutes of stirring
at 0°C and then 6 hours at room temperature, the reaction mixture is diluted with ethyl
acetate, and then neutralized with cold 1N HCI (0-4°C), washed with water, dried
(Na2SO4), filtered and concentrated, and purified on silica gel (ethyl acetate - toluene) to
give 62 (5.86 g, 87%).
LC-MS m/z 976.3 [(M + Na)+]. TR = 27.6/27.8 min
Preparation of (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-idopyranosyl-
uronate)-(1?4)-2-azido-3-O-benzyl-6-O-ferf-butvldiphenvlsilyl-2-deoxy-a,B--D-gluco-
pvranose trichloroacetimidate (No. 63)
To a solution of compound 62 (6.5 g, 6.81 mmol) in dichloromethane (140 mL)
and in the presence of powdered 4 A molecular sieves (7 g), is added, under argon,
caesium carbonate (Cs2C03) (3.5 g, 1.6 molar equivalents), followed by addition, at
0°C, of trichloroacetonitrile (CCI3CN) (3.4 mL, 5.0 molar equivalents). After stirring for
15 minutes 0°C and then for 5 hours at room temperature, the reaction mixture is
filtered and then concentrated. The residue is purified on silica gel (1/4 ethyl
acetate/toluene + 0.1% triethylamine) to give 63 (6.33 g, 85%).
LC-MS m/z 1119.1 [(M + Na)+]. TR = 31.2 min
Preparation of methyl (methyl 2-O-acetyl-3-O-benzyl-4-O-levulinoyl-a-L-
idopvranosvluronate)-(1?4)-(2-aziclo-3-O-benzyl-6-O-tert-butvldiphenvlsilvl-2-deoxy--D-
D-qlucopyranosvl)-(1?4Hmethvl_____2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronateV
(1?4)-3-O-benzyl-2-benzyloxycarbonylamino-6-O-ferf-butvldiphenvlsilvl-2-deoxy-a-D-
qlucopyranoside (No. 64)
A mixture of the glycosyl acceptor 58 (8.80 g, 9.00 mmol), imidate 63 (6.58 g,
6.00 mmol) and powdered 4 A molecular sieves (4.50 g) in dichloromethane (210 mL) is
stirred under an argon atmosphere for 1 hour at 25°C. The reaction mixture is cooled to
-20°C and a 1 M solution of terf-butyldimethylsilyl triflate in dichloromethane (900 uL) is
added. After 1 hour 20 minutes, the reaction medium is neutralized by addition of solid
sodium hydrogen carbonate. After filtering through Celite® and concentrating, the
residue obtained is chromatographed on a Sephadex® LH-20 column (190 x 3.2 cm,
1/1 dichloromethane/ethanol) to give 8.26 g of compound 64.
Rf = 0.30, silica gel, 2/1 cyclohexane/ethyl acetate
Preparation of methyl (methyl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-
(1?4)-(2-azido-3-O-benzyl-6-O-tert-butvldiphenvlsilvl-2-deoxy-a-D-qlucopvranosyl)-
(1?4)-(methyl 2-O-acetyl-3-O-benzyl-a-L-idopvranosvluronate)-(1?4)-3-O-benzyl-2-
benzyloxycarbonylamino-6-O-fe/t-butvldiphenvlsilvl-2-cleoxy-a-D-alucopvranoside (No.
65)
Compound 64 (8.26 g, 4.31 mmol) is converted into compound 65 (6.41 g)
according to the same procedure as that described for the synthesis of 58.
Rf = 0.34, silica gel, 2/1 cyclohexane/ethyl acetate
Preparation of methyl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-
idopvranosvluronate)-(1?4))-(2-azido-3-O-benzyl-6-O-te/t-butvldiphenvlsilyl-2-deoxy--D-
D-qlucopyranosvl)-(1?4Hmethvl_____2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-
(1?4)-(2-azido-3-O-benzyl-6-O-fe/t-butvldiphenvlsilvl-2-deoxy-g--D-qlucopyranosyll)-
(1?4)-(methyl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-(1?4)-3-O-benzyl-2-
benzyloxycarbonylamino-6-O-fe/t-butvldiphenvlsilyl-2-deoxy-a-D-qlucopyranoside (No.
66)
A mixture of the glycosyl acceptor 65 (7.42 g, 4.09 mmol), imidate 63 (6.73 g,
6.1 mmol), and powdered 4 A molecular sieves (4.60 g) in dichloromethane (215 ml_) is
stirred under an argon atmosphere for 1 hour at 25°C. The reaction mixture is cooled to
-20°C and a 1 M solution of tert-butyldimethylsilyl triflate in dichloromethane (920 uL) is
added. After 1 hour 30 minutes, the reaction medium is neutralized by addition of solid
sodium hydrogen carbonate. After filtering through Celite®, the reaction medium is
diluted with dichloromethane (800 mL). The organic phase is washed successively with
2% sodium hydrogen carbonate solution, with water and then dried over sodium sulfate,
filtered and evaporated to dryness. The residue obtained is purified by chromatography
on a Sephadex® LH-20 column (190 x 3.2 cm, 1/1 dichloromethane/ethanol), followed
by chromatography on a column of silica gel (6/1 toluene/ethyl acetate) to give 6.13 g of
compound 66.
Rf = 0.46, silica gel, 4/1 toluene/ethyl acetate
Preparation of methyl (methyl 2-O-acetyl-3-O-benzyl-a-L-idopvranosvluronate)-
(1?4))-(2-azido-3-O-benzyl-6-O-ferf-butvldiphenvlsilvl-2-deoxy-a-D-qlucopyranosyll)-
(1?4)-(methyl 2-O-acetyl-3-O-benzyl--D-i_-idopvranosvluronateM1 -»4)-(2-azido-3-O-
benzyl-6-O-teff-butvldiphenvlsilvl-2-deoxy-a-D-qlucopyranosyll)-(1?4))-(methyl_____2-O-
acetyl-3-O-benzyl-a-L-idopvranosvluronate)-(1?4)-3-O-benzyl-2-benzyloxycarbonyl-
amino-6-O-ferf-butvldiphenvlsilvl-2-deoxy-a-D-qlucopvranoside (No. 67)
Compound 66 (7.14 g, 2.59 mmol) is converted into compound 67 (6.07 g)
according to the same procedure as that described for the preparation of compound 58.
Rf = 0.37, silica gel, 2/1 cyclohexane/ethyl acetate

Preparation of (methyl 2-O-acetyl-3,4-di-O-benzyl-a-L-idopvranosvluronate)-
(1?4)-1 -O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qlucopvranose (No. 68)
To a solution of 36 (5.05 g, 6.3 mmol) in a 1/1 mixture of
methanol/tetrahydrofuran (76 mL) is added [tBu2SnCI(OH)]2 (0.25 g, 0.14 molar
equivalent) prepared according to A. Orita et al., Chem. Eur. J. (2001) 7, 3321. After
stirring at room temperature for 72 hours, the reaction mixture is concentrated under
vacuum and then purified by chromatography to give compound 68 (2.89 g, 64%).
LC-MS m/z 772.4 [(M + Na)+]. TR = 10.23 min
Preparation of (methyl 2-O-acetyl-3,4-di-O-benzyl-a-L-idopyranosvluronate)-
(1?4)-1-O-acetyl-2-azido-3-O-benzyl-6-O-ferf-butyldiphenylsilvl-2-cleoxy-a-D-gluco-
pyranose (No. 69)
Compound 68 (2.89 g, 3.86 mmol) is dissolved in dichloromethane (31 mL).
Triethylamine (1.3 mL, 2.5 molar equivalents), 4-dimethylaminopyridine (0.235 g,
0.5 molar equivalent), and terf-butyldiphenylsilyl chloride (2 mL, 2 molar equivalents) are
successively added at 0°C under argon. After magnetic stirring for 3 hours, the reaction
medium is diluted with dichloromethane, washed with aqueous 10% potassium
hydrogen sulfate solution, with water, dried (Na2SO4), filtered and concentrated. The
residue obtained is purified on silica (ethyl acetate/cyclohexane) to give 69 (3.4 g, 90%).
LC-MS m/z 1010.6 [(M + Na)+]. TR = 13.10 min
Preparation of (methyl 2-O-acetyl-3,4-di-O-benzyl-a-L-idopyranosvluronate)-
(1?4)-2-azido-3-O-benzyl-6-O-terf-butvldiphenvlsilvl-2-deoxy-a.B--D-glucopyranose (No.
70)
To a solution of compound 69 (3.44 g, 3.48 mmol) in diethyl ether (35 mL) is
added, at 0°C, benzylamine (BnNH2) (14.5 mL, 38 molar equivalents). After stirring for
8 hours at room temperature, the reaction mixture is placed at -18°C for 16 hours, and
then again for 2.5 hours at room temperature. The medium is then diluted with ethyl
acetate, and then neutralized with cold 1N HCI (0-4°C), washed with water, dried
(Na2SO4), filtered and concentrated, and purified on silica gel (ethyl acetate/cyclohexane
15/85) to give 70 (3.83 g, 91%).
LC-MS m/z 963.6 [(M + NH4)+]. TR = 12.37, 12.47 min
Preparation of (methyl 2-O-acetyl-3.4-di-O-benzyl-a-L-idopyranosvluronate)-
(1?4)-2-azido-3-O-benzyl-6-O-fe/f-butvldiphenvlsilvl-2-deoxy-a,B--D-qlucopyranose
trichloroacetimidate (No. 71)
To a solution of compound 70 (2.99 g, 3.16 mmol) in dichloromethane (60 mL)
and in the presence of powdered 4 A molecular sieves (3 g), are added, at 0°C under
argon, caesium carbonate (Cs2C03) (1.6 g, 1.6 molar equivalents) and then
trichloroacetonitrile (CCI3CN) (1.6 mL, 5.0 molar equivalents). After 20 minutes of
stirring at 0GC, and 7 hours at room temperature, storing at -18°C for 16 hours, and then
magnetic stirring for 8 hours at room temperature, storing at -18°C for 16 hours, and
finally magnetic stirring for 1 hour at room temperature, the reaction mixture is filtered
and then concentrated. The residue is purified on silica gel (15/85 ethyl acetate/cyclohexane
+ 0.1% triethylamine) to give 71 (2.69 g, 78%).
LC-MS m/z 1113.4 [(M + Na)+]. TR = 14.58 min
Preparation of methyl (methyl 2-O-acetyl-3,4-di-O-benzyl-a-L-idopyranosvl-
uronate)-(1?4)-(2-azido-3-O-benzyl-6-O-teAf-butvldiphenvlsilyl-2-deoxy-a-D-gluco-
pyranosylHI -»4)-f (methyl 2-O-acetyl-3-O-benzyl-g-L-idopyranosvluronate)-(1?4)-(2-
azido-3-O-benzyl-6-O-terf-butvldiphenvlsilvl-2-deoxy-g--D-glucopvranosvl)-(1?4Yl2-
(methyl 2-O-acetyl-3-O-benzyl-g-L-idopvranosvluronate)-(1?4))-3-O-benzyl-2-benzyl-
oxycarbonylamino-6-O-fe/t-butvldiphenvlsilvl-2-deoxy-a-D-glucopvranoside (No. 72)
A mixture of the glycosyl acceptor 67 (3.50 g, 1.32 mmol), imidate 71 (2.16 g,
1.98 mmol) and powdered 4 A molecular sieves (1.48 g) in dichloromethane (69 ml_) is
stirred under an argon atmosphere for 1 hour at room temperature. The reaction
mixture is cooled to -20°C and a 1 M solution of te/f-butyldimethylsilyl triflate in
dichloromethane (297 uL) is added. After 2 hours 30 minutes, the reaction medium is
neutralized by addition of solid sodium hydrogen carbonate. After filtering through
Celite®, the reaction medium is diluted with dichloromethane (400 ml_). The organic
phase is washed successively with 2% sodium hydrogen carbonate solution, with water
and then dried over sodium sulfate, filtered and then evaporated to dryness. The
residue obtained is purified by chromatography on a Sephadex® LH-20 column (190 x
3.2 cm, 1/1 dichloromethane/ethanol), followed by chromatography on a column of silica
gel (cyclohexane/ethyl acetate 4/1) to give 3.04 g of compound 72.
Rf = 0.30, silica gel, 3/1 cyclohexane/ethyl acetate
Preparation of methyl (methyl 3.4-di-O-benzyl-g-i_-idopvranosvluronate)-(1?4)-
(2-azido-3-O-benzyl-6-O-tert-butvldiphenvlsilvl-2-deoxy-a-D-glucopvranosvl)-(1?4)-
r(methyl_____3-O-benzyl-a-L-idopvranosvluronate)-(1?4))-(2-azido-3-O-benzyl-6-O-fert-
butvldiphenvlsilvl-2-deoxy-a-D-glucopyranosvl)-(1?4)l7-(methvl________3-O-benzyl-o-L-
idopvranosyluronate)-(1?4))-3-O-benzyl-2-benzyloxycarbonylamino-6-O-terf-
butvldiphenvlsilvl-2-deoxy-g--D-glucopyranoside (No. 73)
To a solution of compound 72 (2.23 g, 0.623 mmol) in a 2/3
dichloromethane/methanol mixture (187 ml_) containing 3A sieves (78 mg) is added,
under an argon atmosphere and at 0°C, a 1 M solution of sodium methoxide in
methanol (99.7 uL). After 24 hours at room temperature, the reaction medium is
neutralized with Dowex AG 50 WX4 H+ resin. After filtering and concentrating, the
residue is chromatographed on a Sephadex® LH-20 column (120 x 3 cm, 1/1
dichloromethane/ethanol), followed by flash chromatography on a column of silica gel
(1/0-^66/34 cyclohexane/ethyl acetate) to give 1.80 g of compound 73.
Rf = 0.38, silica gel, 3/1 cyclohexane/ethyl acetate
Preparation of methyl (methyl 3,4-di-O-benzyl-a-L-idopvranosyluronate)-(1?4)-
(2-azido-3-O-benzyl-2-deoxy-a-D-qlucopvranosyl)-(1?4)-r(methyl______3-O-benzyl-a-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-a-D-glucopyranosvl)-( 1?4)V
(methyl 3-O-benzyl-O-L-idopvranosvluronate)-(1?4)-3-O-benzyl-2-benzyloxycarbonyl-
amino-2-deoxy-a-D-qlucopyranoside (No. 74)
To a solution of compound 73 (44,0 mg, 0,013 mmol) in methanol (1,7 ml_) is
added ammonium fluoride (38.0 mg, 1.03 mmol). After 140 hours of stirring at room
temperature, the reaction mixture is applied to a Sephadex® LH-20 column (95 x 2 cm,
1/1 dichloromethane/ethanol), followed by flash chromatography on a column of silica
gel (3/2 toluene/acetone) to give compound 74 (24.2 mg).
Rf = 0.46, silica gel, 3/2 toluene/acetone
Preparation of methyl (methyl 3,4-di-O-benzyl-2-O-triethvlammonium sulfonato-
a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-triethvlammonium
sulfonato-a-D-glucopvranosvlH(1?4Hmethyl_______3-O-benzyl-2-O-triethvlammonium
sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-triethyl-
ammonium sulfonato-a-D-glucopyranosvl)1?-(1?4)-(methyl 3-O-benzyl-2-O-triethvl-
ammonium sulfonato-a-L-idopvranosvluronate)-(1?4)-3-O-benzyl-2-benzyloxycarbonyl-
amino-2-deoxy-6-O-triethvlammonium sulfonato-a-D-glucopyranoside (No. 75)
Compound 74 (24.2 mg, 9.96 umol) is dried by codistillation of N,N-
dimethylformamide (3 x 1.0 mL) and is then dissolved in A/,/V-dimethylformamide
(1,0 mL). To this solution is added triethylamine-sulfur trioxide complex (72.2 mg,
0.4 mmol). The mixture is stirred for 16 hours at 55°C sheltered from light and the
excess reagent is then destroyed with methanol (16.2 uL). The reaction medium is
applied to a Sephadex® LH-20 column (120 x 3 cm) eluted with a 1/1 mixture of
dichloromethane/ethanol to give compound 75 (29.0 mg).
Rf = 0.26, silica gel, ethyl acetate/pyridine/acetic acid/water 28/16/3.8/9.
Preparation of methyl (lithium 3,4-di-O-benzyl-2-O-lithium sulfonato-a-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-a-D-
glucopyranosvlW1?4)-(lithium 3-O-benzyl-2-O-Hthium sulfonato-O-L-idopyranosyl-
uronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-lithium_________sulfonato-a-D-gluco-
pyranosvM7-(1?4Hlithium 3-O-benzyl-2-O-lithium sulfonato-o-L-idopyranosvluronate)-
(1?4)-3-O-benzyl-2-benzyloxycarbonylamino-2-deoxy-6-O-lithium sulfonato-a-D-gluco-
pyranoside (No. 76)
To a solution of compound 75 (78.5 mg, 30.1 umol) in a 1/1
tetrahydrofuran/methanol mixture (3.2 mL) is added, at 0°C, a 0.7 M solution of lithium
hydroxide in water (440 uL). After 1 hour at 0°C and then 16 hours at room temperature,
the solution is applied to a Sephadex® LH-20 column (3 x 120 cm) eluted with a 50/50/1
dichloromethane/ethanol/water mixture to give compound 76 (60.3 mg).
[a]D 13.1° (c 1.0; MeOH)
Preparation of methyl (sodium 2-O-sodium sulfonato-a-L-idopvranosvluronate)-
(1?4H2-amino-2-deoxy-6-O-sodium sulfonato-g--D-qlucopyranosvO-rd—>4)-(sodium 2-
O-sodium sulfonato-a-L-idopvranosvluronate)-(1^4M2-amino-2-deoxy-6-O-sodium
sulfonato-g--D-glucopyranosvDVd—>4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosvl-
uronate)-(1?4)-2-amino-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranoside (No. 77)
To a solution of compound 76 (19.3 mg, 6.16 umol) in a 1/1 terf-butanol/water
mixture (1.2 mL) are successively added ammonium formate (50.8 mg) and Pd/C 10%
(125 mg). After 4 hours at room temperature, the reaction mixture is filtered (Millipore®
LSWP 5 urn filter) and then applied to a column of fine Sephadex® G-25 gel (95 x 2 cm)
eluted with aqueous 0.2 M NaCI solution. The fractions containing the expected product
are pooled and applied to a column of fine Sephadex® G-25 (95 x 2 cm) eluted with
water, to give 11.2 mg of compound 77.
Mass: "ESI" method, negative mode: theoretical mass = 2285.47; experimental
mass: 2197.20 + 0.34 a.m.u. (iduronic acids observed in COOH form)
Preparation of methyl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-o-L-
idopvranosvluronate)-(1?4))-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qluco-
pvranosvl)-(1?4Hmethyl 2-O-acetyl-3-O-benzyl-a-L-idopvranosvluronate)-(1?4)-(2-
azido-3-O-benzyl-6-O-tert-butyldiphenvlsilvl-2-deoxy-a-D-glucopyranosvl)-(1?4))-(methvl
2-O-acetyl-3-O-benzyl-g-L-idopvranosvluronate)-(1?4)-3-O-benzyl-2-benzyloxy-
carbonylamino-6-O-re/t-butvldiphenvlsilvl-2-deoxy-g--D-glucopyranoside (No. 78)
A mixture of the glycosyl acceptor 65 (1.82 g, 1.00 mmol), imidate 27 (1.35 g,
1.50 mmol) and powdered 4 A molecular sieves (1.12 g) in dichloromethane (52 mL) is
stirred under an argon atmosphere for 1 hour at 25°C. The reaction mixture is cooled to
-20°C and a 1 M solution of terf-butyldimethylsilyl triflate in dichloromethane (224 uL) is
then added. After 50 minutes, the reaction medium is neutralized by addition of solid
sodium hydrogen carbonate. After filtering through Celite®, the reaction medium is
diluted with dichloromethane (500 mL). The organic phase is washed successively with
aqueous 2% sodium hydrogen carbonate solution, with water and then dried over
sodium sulfate, filtered and then evaporated to dryness. The residue obtained is
chromatographed on a column (Sephadex® LH-20, 190 x 3.2 cm, 1/1
dichloromethane/ethanol), followed by flash chromatography on a column of silica gel
(1/0—>3/1 toluene/acetone) to give 1.57 g of compound 78.
Rf = 0.47, silica gel, 3/1 toluene/acetone
Preparation of methyl (methyl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-
(1?4H6-O-acefyl-2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosvlH1^4Hrnethvl 2-O-
acetyl-3-O-benzyl-a-L-idopyranosvluronate)-(1?4)-(2-azido-3-O-benzyl-6-O-te/-f-butvl-
diphenvlsilvl-2-deoxy-a-D-qlucopyranosvl)-(1?4)-(methvl 2-O-acetyl-3-O-benzyl-a-L-
idopvranosvluronate)-(1?4))-3-O-benzyl-2-benzyloxycarbonylamino-6-O-teAf-butvl-
diphenvlsilvl-2-deoxy-a-D-qlucopyranoside (No. 79)
Compound 78 (1.56 g, 0.61 mmol) is converted into compound 79 (1.44 g)
according to the same procedure as that described for the preparation of compound 58.
Rf = 0.52, silica gel, 1/1 toluene/acetone
Preparation of methyl (methyl 2-O-acetyl-3,4-di-O-benzyl-a-L-idopyranosvl-
uronateV(1?4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosyll)-(1?4)-
(methyl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-(1?4)-(6-O-acetyl-2-azido-3-
0-benzyl-2-deoxy-a-D-qlucopyranosyll)-(1?4)-(methvl_______2- O-acetyl-3- O-benzyl-a-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-6-O-teAf-butvldiphenvlsilvl-2-deoxy--D-
D-glucopvranosyl)-(1?4)-(methvl_____2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-
(1?4)-3-O-benzyl-2-benzyloxycarbonylamino-6-O-teAf-butvldiphenvlsilyl-2-deoxy-a-D-
qlucopyranoside (No. 80)
A mixture of the glycosyl acceptor 79 (1.0 g, 0.41 mmol), imidate 38 (543 mg,
0.61 mmol) and powdered 4 A molecular sieves (455 mg) in dichloromethane (21 mL) is
stirred under an argon atmosphere for 1 hour at room temperature. The reaction
mixture is cooled to -20°C and a 1 M solution of fe/f-butyldimethylsilyl triflate in
dichloromethane (91.0 uL) is added. After 1 hour 10 minutes, the reaction medium is
neutralized by addition of solid sodium hydrogen carbonate. After filtering through
Celite®, the reaction medium is diluted with dichloromethane (250 mL). The organic
phase is washed successively with aqueous 2% sodium hydrogen carbonate solution,
with water and then dried over sodium sulfate, filtered and then evaporated to dryness.
The residue obtained is purified by chromatography on a Sephadex® LH-20 column
(190 x 3.2 cm, 1/1 dichloromethane/ethanol), followed by flash chromatography on a
column of silica gel (1/0—>0/1 toluene/ethyl acetate) to give compound 80 (822.4 mg).
Rf = 0.44, silica gel, 3/1 toluene/ethyl acetate
Preparation of methyl (methyl 3.4-di-O-benzyl-g-L-idopyranosyluronate)-(1?4)-
(2-azido-3-O-benzyl-2-deoxy-g--D-qlucopyranosylD-M->4Hmethvl______3-O-benzyl-a-L-
idopvranosyluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosyll)-( 1?4)-
(methyl______3-O-benzyl-a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-6-O-te/t-
butvldiphenvlsilvl-2-deoxy-a-D-qlucopyranosyllH1?4)-(methy[_________3-O-benzyl-a-L-
idopvranosvluronateH1?4)-3-O-benzyl-2-benzyloxycarbonylamino-6-O-terf-
butvldiphenvlsilvl-2-deoxy-a-D-qlucopvranoside (No. 81)
Compound 80 (820 mg, 0.257 mmol) is saponified according to the method used
for the preparation of compound 73, to give the octasaccharide 81 (593.8 mg) after
flash chromatography on a column of silica gel (1/0—>0/1 toluene/acetone).
Rf = 0.57, silica gel, 2/1 toluene/acetone
Preparation of methyl (methyl 3,4-di-O-benzyl-2-O-triethylammonium sulfonato-
a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-triethylammonium
sulfonato-g--D-glucopyranosvO-(1?4)-(methvl_______3-O-benzyl-2-O-triethylammonium
sulfonato-a-L-idopvranosvluronate)-(1?4))-(2-azido-3-O-benzyl-2-deoxy-6-O-triethvl-
ammonium sulfonato-g--D-glucopvranosvl)-(1?4)-(methyl 3-O-benzyl-2-O- triethvl-
ammonium sulfonato-g-L-idopvranosvluronateM1?4)-(2-azido-3-O-benzyl-6-O-ferf-
butvldiphenvlsilvl-2-deoxy-g--D-glucopvranosvO-(1?4)-(methvl 3-O-benzyl-2-O- triethyl-
ammonium sulfonato-g-L-idopyranosvluronate)-(1?4)-3-O-benzyl-2-benzyloxycarbonyl-
amino-6-O-ferf-butvldiphenvlsilvl-2-deoxy-g--D-glucopvranoside (No. 82)
Compound 81 (130 mg, 44.3 umol) is converted into compound 82 (164.1 mg)
according to the same procedure as that described for the preparation of compound 75.
Mass: "ESI" method, negative mode: theoretical mass = 4024.90; experimental
mass: 4024
Preparation of methyl (methyl 2-O-ammonium sulfonato-3,4-di-O-benzyl-g-L-
idopvranosvluronate)-(1?4)-(6-O-ammonium sulfonato-2-azido-3-O-benzyl-2-deoxy-g-
D-glucopyranosvl)-(1?4)-(methyl______2-O-ammonium______sulfonato-3-O-benzyl-g-L-
idopyranosyluronateVd -»4)-( 6-O-ammonium sulfonato-g-2-azido-3-O-benzyl-2-deoxy-
D-glucopyranosvl)-(1?4)-(methyl______2-O-ammonium______sulfonato-3-O-benzyl-g-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-g--D-qlucopyranosvl)-(1?4)-
(methyl 2-O-ammonium sulfonato-3-O-benzyl-g-L-idopvranosyluronate)-(1?4)-3-O-
benzyl-2-benzyloxycarbonylamino-2-deoxy-g--D-qlucopvranoside (No. 83)
To a solution of compound 82 (91.8 mg, 22.8 umol) in methanol (3.0 mL) is
added ammonium fluoride (67.6 mg, 1.82 mmol). After 48 hours of stirring at 55°C, the
reaction mixture is applied to a Sephadex® LH-20 column (120 x 3 cm, 1/1
dichloromethane/ethanol), followed by flash chromatography on a column of silica gel
(95/5 methanol/water) to give compound 83 (62.7 mg).
Rf = 0.32, silica gel, 17/9/2.2/5 ethyl acetate/pyridine/acetic acid/water.
Preparation of methyl (lithium 3,4-di-O-benzyl-2-O-lithium sulfonato-g-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-g--D-
qlucopyranosvl)-(1?4)-(lithium 3-O-benzyl-2-O-lithium sulfonato-g-L-idopyranosyl-
uronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-g--D-glucopyranosyl)-
(1?4)-(lithium 3-O-benzyl-2-O-lithium sulfonato-g-L-idopvranosyluronate)-(1?4)-(2-
azido-3-O-benzyl-2-deoxy-o--D-glucopvranosylH1?4Hlithium 3-O-benzyl-2-O-lithium
sulfonato-a-L-idopvranosvluronate)-(1?4)-3-O-benzyl-2-benzyloxycarbonylamino-2-
deoxy-g--D-qlucopyranoside (No. 84)
To a solution of compound 83 (86 mg, 24.2 umol) in a 1/1
tetrahydrofuran/methanol mixture (3.9 mL) is added, at 0°C, a 1 M solution of lithium
hydroxide in water (780 uL). After 1 hour at 0°C and then 16 hours at room temperature,
the solution is applied to a column of Sephadex® LH-20 gel (95 x 2 cm) eluted with a
95/5 methanol/water mixture to give compound 84 (65.3 mg).
Rf = 0.27, silica gel, 17/9/2.2/5 ethyl acetate/pyridine/acetic acid/water.
Preparation of methyl (sodium 2-O-sodium sulfonato-g-L-idopyranosvluronate)-
(1?4)-(2-amino-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosvl)-(1?4)-(sodium 2-
O-sodium sulfonato-a-L-idopvranosyluronate)-(1?4M2-amino-2-deoxy-6-O-sodium
sulfonato-a-D-qlucopyranosyll)-(1?4Hsodium 2-O-sodium sulfonato-a-L-idopyranosvl-
uronate)-(1?4)-(2-amino-2-deoxy-O--D-qlucopvranosyl)-(1?4)-(sodium 2-O-sodium
sulfonato-g-L-idopvranosvluronate)-(1?4)-2-amino-2-deoxy-g--D-qlucopyranoside (No.
85)
Compound 84 (95.0 mg, 32.3 umol) is treated according to the same procedure
as that described for the preparation of compound 77, to give compound 85 (47.3 mg).
Mass: "ESI" method, negative mode: theoretical mass = 2081.38; experimental
mass: 1993.15 ± 0.11 a.m.u. (iduronic acids observed in COOH form).
Preparation of methyl (methyl 3,4-di-O-benzyl-a-L-idopvranosyluronate)-(1?4)-
(2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosyn-(1?4))-(methvl_______3-O-benzyl-g-L-
idopvranosvluronate)-(1?4))-(2-azido-3-O-benzyl-2-deoxy-a-D-glucopyranosyl)-(1?4))-
(methyl______3-O-benzyl-a-L-idopyranosvluronate)-(1?4)-(2-azido-3-c3-benzyl-6-O-fert-
butvldiphenvlsilyl-2-deoxy-a-D-qlucopyranosvl)-(1?4)-(methyl_________3-O-benzyl-a-L-
idopyrariosvluronate)-(1?4)-3-O-benzyl-2-benzyloxycarbonylamino-2-de6xv-a-D-gluco-
pyranoside (No. 86)
To a solution of compound 81 (50 mg, 0.017 mmol) in methanol (2.2 mL) is
added ammonium fluoride (25 mg, 0.68 mmol). After 7 hours of stirring at room
temperature and then 16 hours at 20°C, the reaction mixture is diluted with
dichloromethane. The organic phase is washed with water, dried over sodium sulfate,
filtered and then evaporated to dryness. The residue is purified by flash chromatography
on a column of silica gel (100/0->82/18 toluene/methanol) to give 27.7 mg of compound
86.
Rf = 0.16, silica gel, 85/15 toluene/methanol
Preparation of methyl (methyl 3,4-di-O-benzyl-2-O-triethvlammonium sulfonato-
a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-triethvlammonium
sulfonato-a-D-qlucopyranosvl)-(1?4)-(methvl_______3-O-benzyl-2-O-triethylammonium
sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-triethvl-
ammonium sulfonato-O--D-qlucopyranosyll)-(1?4)-(methyl 3-O-benzyl-2-O-triethvl-
ammonium sulfonato-O-L-idopvranosvluronateH1?4)-(2-azido-3-O-benzyl-6-O-terf-
butvldiphenvlsilyl-2-deoxy-a-D-qlucopyranosyll)-(1-*4Mmethvl 3-O-benzyl-2-O-triethvl-
ammonium sulfonato-O-L-idopvranosvluronate)-(1?4)-3-O-benzyl-2-benzyloxycarbonyl-
amino-2-deoxy-6-O-triethvlammonium sulfonato-O--D-glucopyranoside (No. 87)
Compound 86 (94 mg, 34.8 umol) is treated according to the same procedure as
that described for the preparation of compound 75, to give compound 87 (131.9 mg).
Rf = 0.37, silica gel, 17/9/2.2/5 ethyl acetate/pyridine/acetic acid/water.
Preparation of methyl (methyl 2-O-ammonium sulfonato-3,4-di-O-benzyl-a-L-
idopvranosvluronate)-(1?4)-(6-O-ammonium sulfonato-2-azido-3-O-benzyl-2-deoxy-o-
D-glucopvranosvl)-(lH>4Mmethvl______2-O-ammonium______sulfonato-3-O-benzyl-a-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-O--D-qlucopyranosvl)-(1?4))-
(methvl 2-O-ammonium sulfonato-3-O-benzyl-O-i_-idopvranosvluronate)-(1?4H2-azido-
3-O-benzyl-2-deoxy-a-D-qlucopyranosyll)-(1?4)-(methyl 3-O-benzyl-a-L-idopvranosvl-
uronate)-(1?4)-6-O-ammonium sulfonato-3-O-benzyl-2-benzyloxycarbonylamino-2-
deoxy-O--D-qlucopyranoside (No. 88)
Compound 87 (130 mg, 32.8 umol) is treated according to the same procedure
as that described for the preparation of compound 74, to give compound 88 (102.0 mg).
Rf = 0.33, silica gel, 17/9/2.2/5 ethyl acetate/pyridine/acetic acid/water.
Preparation of methyl (lithium 3,4-di-O-benzyl-2-O-lithium sulfonato-O-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-a-D-
qlucopyranosyll)-(1?4Hlithium 3-O-benzy[-2-O-lithium sulfonato-a-L-idopyranosyl-
uronateH1^4H2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-a-D-qlucopyranosylO-
(1—»4)-(lithium 3-O-benzyl-2-O-lithium sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-
azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosyll)-(1?4Hlithium 3-O-benzyl-2-O-lithium
sulfonato-a-L-idopvranosvluronate)-(1?4)-3-O-benzyl-2-benzyloxycarbonylamino-2-
deoxy-6-O-lithium sulfonato-a-D-glucopyranoside (No. 89)
Compound 88 (100 mg, 26.8 umol) is treated according to the same procedure
as that described for the preparation of compound 76, to give crude compound 89,
which is used without further purification in the following step.
Rf = 0.25, silica gel, 17/9/2.2/5 ethyl acetate/pyridine/acetic acid/water.
Preparation of methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-
(1?4)-(2-amino-2-deoxy-6-O-sodium sulfonato-a-D-qlucopyranosvP-(1-»4Hsodium 2-
O-sodium sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-amino-2-deoxy-6-O-sodium
sulfonato-a-D-qlucopyranosvlM1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronate)-(1-H>4H2-amino-2-deoxy-a-D-qlucopyranosyllH1-»4)-(sodium 2-O-sodium
sulfonato-a-L-idopvranosvluronate)-(1?4)-2-amino-2-deoxy-6-O-sodium sulfonato--D-p-
qlucopyranoside (No. 90)
Crude compound 89 (75 mg, 24.8 umol) is treated according to the same
procedure as that described for the preparation of compound 77 to give compound 90
(48.2 mg).
Mass: "ESI" method, negative mode: theoretical mass = 2183.42; experimental
mass: 2095.07 ± 0.24 a.m.u. (iduronic acids observed in COOH form)
Preparation of methyl (methyl 2-O-acetv[-3-O-benzyl-4-O-leyulinoyl-o-L-
idopvranosvluronateV(1?4)H6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-O--D-qluco-
pyranosylHI ->4Hmethvl 2-O-acetyl-3-O-benzyl-a-L-idopvranosyluronate)-(1?4)-3-O-
benzyl-6-O-terf-butvldiphenvlsilvl-2-deoxy-2-r(benzyloxy)carbonvnamino-a-D-qluco-
pyranoside (No. 91)
A solution of ferf-butyldimethylsilyl triflate in dichloromethane (1M, 0.15 mol per
mole of imidate) is added, under argon and at -20°C, to a solution of the imidate 27
(3.26 g, 3.62 mmol) and of the glycosyl acceptor 58 (3.22 g, 3.29 mmol) in
dichloromethane (115 mL) in the presence of 4 A molecular sieves (2.5 g). After 1 hour
40 minutes at -20°C (TLC), solid sodium hydrogen carbonate is added. After filtering,
washing with aqueous 2% sodium hydrogen carbonate solution, with aqueous sodium
chloride solution, followed by drying (Na2SO4) and evaporating to dryness, the residue is
purified on silica gel (toluene-acetone) to give 91 (4.66 g, 83%).
Rf = 0.49 (7/3 toluene/acetone).
Preparation of methyl (methyl 2-O-acetyl-3-O-benzyl-g-i_-idopvranosvluronate)-
(1?4M6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-g--D-qlucopyranosyll)-(1?4)-(methvl 2-O-
acetyl-3-O-benzyl-a-L-idopvranosvluronate)-(1?4)-3-O-benzyl-6-O-fe/t-butvldiphenvl-
silvl-2-deoxy-2-r(benzyloxy)carbonvnamino-a-D-qlucopyranoside (No. 92)
To a solution of compound 91 (4.66 g, 2.71 mmol) in a 1/2 mixture of
toluene/ethanol (542 mL) is added hydrazine acetate (1.2 g, 5 molar equivalents). After
magnetic stirring for 2 hours, the same amount of reagent is added and after 2 hours,
the mixture is concentrated under vacuum and then purified on silica gel (toluene-
acetone) to give compound 92 (3.83 g, 87%).
Rf = 0.42 (3/1 toluene-acetone).
Preparation of methyl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-
idopvranosvluronate)-(1?4))-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qluco-
pvranosvl)-(1?4)-(methvl 2-O-acetyl-3-O-benzyl-O-L-idopvranosvluronateM1 -»4)-(6-O-
acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosyll)-(1?4)-(methvl 2-O-acetyl-3-O-
benzyl-a-L-idopvranosvluronate)-(1?4)-3-O-benzyl-6-O-ferf-butvldiphenvlsilvl-2-deoxy-
2-r(benzyloxy)carbonyllamino-a-D-qlucopyranoside (No. 93)
A solution of terf-butyldimethylsilyl triflate in dichloromethane (1M, 0.15 mol per
mole of imidate) is added, under argon and at -20°C, to a solution of the imidate 27
(3.2 g, 3.55 mmol) and of the glycosyl acceptor 92 (3.83 g, 2.36 mmol) in
dichloromethane (106 mL) in the presence of 4 A molecular sieves (1.8 g). After
2.5 hours at -20°C (TLC), solid sodium hydrogen carbonate is added. After filtering,
washing with aqueous 2% sodium hydrogen carbonate solution, with aqueous sodium
chloride solution, drying (Na2SO4) and evaporating to dryness, the residue is purified on
silica gel (ethyl acetate/toluene) to give 93 (3.36 g, 61%).
Rf = 0.47 (1/1 toluene/ethyl acetate).
Preparation of methyl (methyl 2-O-acetyl-3-O-benzyl-o-L-idopyranosvluronate)-
(1?4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-g--D-glucopvranosv0-(1?4Hmethyl 2-O-
acetyl-3-O-benzyl-a-L-idopvranosyluronate)-(1—»4)-(6-O-acetyl-2-azido-3-O-benzyl-2-
deoxy-g--D-qlucopyranosylO-(1?4)-(methvl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvl-
uronate)-(1^-4)-3-O-benzyl-6-O-te/t-butvldiphenvlsilvl-2-deoxy-2-r(benzyloxy)carbonvn-
amino-g--D-glucopvranoside (No. 94)
To a solution of compound 93 (3.36 g, 1.42 mmol) in a 1/2 mixture of
toluene/ethanol (280 mL) is added hydrazine hydrate (0.645 g, 4.93 molar equivalents).
After 2.5 hours of magnetic stirring, the mixture is concentrated under vacuum and then
purified on silica gel to give compound 94 (3.01 g, 96%).
Rf = 0.43 (4/1 toluene/acetone).
Preparation of methyl (methyl 2-O-acetyl-3,4-di-O-benzyl-O-L-idopyranosvl-
uronate)-(1?4)-(2-azido-3-O-benzyl-6-O-terf-butvldiphenvlsilyl-2-deoxy-g--D-gluco-
pyranosvlHI —>4Hmethvl 2-O-acetyl-3-O-benzyl-g-L-idopvranosvluronate)-(1?4)-(6-O-
acetyl-2-azido-3-O-benzyl-2-deoxy-g-p-glucopyranosvl)-(1?4)-(methvl 2-O-acetyl-3-O-
benzyl-g-L-idopvranosvluronate)-(1?4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-
qlucopyranosyll)-(1?4)-(methvl 2-O-acetyl-3-O-benzyl-g-L-idopvranosvluronateH1 -*4)-
3-O-benzyl-6-O-fe/f-butvldiphenvlsilvl-2-deoxy-2-r(benzyloxy)carbonvnamino-g--D-gluco-
pyranoside (No. 95)
A 0.1 M solution of terf-butyldimethylsilyl triflate in dichloromethane (1 mL) is
added, under argon and at -20°C, to a solution of the imidate 71 (0.29 g, 0.3 mmol) and
of the glycosyl acceptor 94 (3.01 g, 1.33 mmol) in dichloromethane (60 mL) in the
presence of 4 A molecular sieves (0.93 g). After magnetic stirring for 10 minutes at
-20°C (TLC), imidate 71 is again added at time intervals of between 10 minutes and
2.5 hours, up to a total amount of 3.19 g. After 20 hours at -20°C (TLC), solid sodium
hydrogen carbonate is added to neutralize, and after filtering, washing with aqueous 2%
sodium hydrogen carbonate solution, with aqueous sodium chloride solution, drying
(Na2SO4) and evaporating to dryness, the residue is purified on silica gel to give 95
(2.58 g, 61%).
Rf = 0.45 (4/1 toluene/acetone).
Preparation of methyl (methyl 3,4-di-O-benzyl-g-L-idopvranosvluronateH1?4)-
(2-azido-3-O-benzyl-6-O-fert-butvldiphenvlsilvl-2-deoxy-g--D-glucopyranosvl)-(1?4)-
(methyl 3-O-benzyl-a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-a-D-
qlucopyranosvlH1?4Hmethvl 3-O-benzyl-a-L-idopvranosvluronateH1?4H2-azido-3-
0-benzyl-2-deoxy-a-D-qlucopyranosyllH1?4Hmethvl 3-O-benzyl-g-L-idopvranosvl-
uronate)-(1^-4)-3-O-benzyl-6-O-terf-butvldiphenvlsilvl-2-deoxy-2-f(benzyloxy)carbonvn-
amino-g--D-qlucopyranoside (No. 96)
To a solution of 95 (1.18 g, 0.37 mmol) in a 3/2 mixture of
methanol/dichloromethane (111 mL) is added a 1M a methanolic solution of sodium
methoxide (1.1 mL), in the presence of 3 A molecular sieves (463 mg). After stirring for
20 hours at room temperature, the mixture is neutralized with H+ Dowex 50WX4 resin.
After filtering and concentrating, the residue is purified on silica gel with a
toluene/acetone mixture to give compound 96 (773 mg, 77%).
Rf = 0.51 (4/1 toluene/acetone).
Preparation of methyl (methyl 3,4-di-O-benzyl-2-O-triethylammonium sulfonato-
q-L-idopvranosvluronateM1?4)-(2-azido-3-O-benzyl-6-O-te/t-butvldiphenvlsilyl-2-
deoxy-O--D-qlucopyranosyllV(1 -»4Mmethyl 3-O-benzyl-2-O-triethylammonium sulfonato-
a-L-idopyranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-triethvlammonium
sulfonato-O--D-qlucopyranosylO-(1?4)-(methvl_______3-O-benzyl-2-O-triethylammonium
sulfonato-O-L-idopvranosvluronateM1?4H2-azido-3-O-benzyl-2-deoxy-6-O-triethvl-
ammonium sulfonato-a-D-qlucopyranosyllH1?4Mmethvl 3-O-benzyl-2-O-triethyl-
ammonium_______sulfonato-a-L-idopyranosvluronate)-(1?4)-3-O-benzyl-6-O-teff-butvl-
diphenvlsilvl-2-deoxy-2-[(benzyloxy)carbonvnamino-O--D-qlucopyranoside (No. 97)
Triethylamine/sulfur trioxide complex (761 mg, 5 mol per hydroxyl function) is
added to a solution in A/,A/-dimethylformamide (11 ml_, 90 L/mol) of compound 96
(324 mg, 120 umol). After 17 hours of magnetic stirring at 55°C sheltered from light,
methanol is added at 0°C and, after stirring for 30 minutes at 0°C and then for
30 minutes at room temperature, the reaction medium is diluted with methanol and then
purified by means of an LH-20 column, using a 1/1 mixture of
methanol/dichloromethane as eluent. The fractions containing the product are then
concentrated under high vacuum and, if necessary, reacted again under the same
conditions. Compound 97 obtained is then used directly in the following step.
Rf = 0.5 (11/7/1.6/4 EtOAc/pyridine/AcOH/H20).
Preparation of methyl (methyl 2-O-ammonium sulfonato-3,4-di-O-benzyl-O-L-
idopyranosyluronateMI -»4M2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosvl)-( 1 -»4V
(methyl 2-O-ammonium sulfonato-3-O-benzyl-O-i_-idopvranosyluronate)-(1?4)-(6-O-
ammonium sulfonato-2-azido-3-O-benzyl-2-deoxy-O--D-qlucopyranosvD-( 1?4)-(methvl 2-
O-ammonium sulfonato-3-O-benzyl-O-L-idopvranosvluronate)-(1?4)-(6-O-ammonium
sulfonato-2-azido-3-O-benzyl-2-deoxy-O--D-qlucopyranosvlM1?4)-(methvl________2-O-
ammonium sulfonato-3-O-benzyl-O-L-idopyranosvluronate)-(1?4)-3-O-benzyl-2-deoxy-
2-r(benzyloxy)carbonvnamino-O--D-qlucopvranoside (No. 98)
To a solution of compound 97 obtained previously in methanol (16 ml_) is added
ammonium fluoride (361 mg, 80 molar equivalents). After magnetic stirring at 55°C for
48 hours, the reaction mixture is purified with the aid of an LH-20 column, using a
75/20/5 mixture of methanol/A/,A/-dimethylformamide/water as eluent. The fractions
containing the product are then concentrated under high vacuum to give compound 98
(410 mg, 95%, 2 steps).
Rf = 0.28 (57/29/7.2/16 EtOAc/pyridine/AcOH/H20).
Preparation of methyl (lithium 3.4-di-O-benzyl-2-O-lithium sulfonato-O-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-O--D-qlucopvranosyl)-(1?4)-
(lithium 3-O-benzyl-2-O-lithium sulfonato-O-L-idopvranosvluronateH1?4)-(2-azido-3-O-
benzyl-2-deoxy-6-O-lithium sulfonato-O--D-alucopvranosvlV( 1 -»4 V(3-O-benzyl-2-O-
lithium sulfonato-g-L-idopvranosvluronateM1?4H2-azido-3-O-benzyl-2-deoxy-6-O-
lithium sulfonato-g--D-qlucopyranosyllH1?4V(lithium 3-O-benzyl-2-O-lithium sulfonato-
a-L-idopvranosvluronateV(1?4)V3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonvnamino-a-D-
qlucopvranoside (No. 99)
To a solution of compound 98 (202 mg, 57 umol) in a 1/1 mixture of
methanol/tetrahydrofuran (9 mL) is added, dropwise and at 0°C, aqueous 1N LiOH
solution (1.8 mL, 7.9 equivalents/methyl ester function). After stirring for 1 hour at 0°C,
and then for 16 hours at room temperature, the reaction medium is purified by means of
an LH-20 column, using a 75/20/5 mixture of methanol/W,W-dimethylformamide/water as
eluent. The fractions containing the product are then concentrated under high vacuum
to give the desired compound 99, which is used directly in the following step.
Rf = 0.41 (57/29/7.2/16 EtOAc/pyridine/AcOH/H20)
Preparation of methyl (lithium 2-O-lithium sulfonato-O-L-idopvranosvluronate)-
(1?4)-(2-amino-2-deoxy-O--D-qlucopyranosyll)-(1?4)-(lithium 2-O-lithium sulfortato-a-L-
idopvranosvluronateH1^4M2-amino-2-deoxy-6-O-lithium________sulfonato-g--D-qluco-
pvranosvlM1?4)-(lithium 2-O-lithium sulfonato-a-L-idopyranosvluronateV1?4)-(2-
, amino-2-deoxy-6-O-lithium sulfonato-o--D-qlucopyranosyll)-(1?4)-(lithium 2-O-lithium
sulfonato-O-L-idopvranosvluronate)-(1?4)-2-amino-2-deoxy-g--D-qlucopvranoside (No.
100)
To a solution in a 1/1 mixture of terf-butanol/water (13 mL) of compound 99
obtained previously are successively added ammonium formate (529 mg, 8.4 mmol)
and then 10% Pd/C (1.24 g). After 4 hours 15 minutes of vigorous stirring at room
temperature, the reaction medium is filtered and partially concentrated under vacuum,
and the solution is then applied to a Sephadex® G-25 column eluted with 0.2 M sodium
chloride. The fractions containing the product are concentrated and desalified using the
same column eluted with water. The fractions containing the product are then
concentrated under high vacuum to give the desired compound 100 (78 mg, 66%,
2 steps).
Mass: "ESI" method, negative mode: theoretical mass = 1992.8; experimental
mass [M - Na - H]: 1971.03 ± 0.10 a.m.u. (iduronic acids observed in COOH form).
Preparation of methyl (methyl 3,4-di-O-benzyl-a-L-idopyranosvluronateV(1?4))-
(2-azido-3-O-benzyl-6-O-fe/-f-butvldipherivlsilvl-2-deoxy-a-D-qlucopyranosyln-(1—»4)-
(methyl 3-O-benzyl-a-L-idopvranosvluronateV(1?4)-(2-azido-3-O-benzyl-2-deoxy-a-D-
qlucopyranosylHI —>4Mmethyl 3-O-benzyl-a-L-idopyranosyluronate)-(1?4M2-azido-3-
0-benzyl-2-deoxy-a-D-qlucopyranosvn-(1—»4V(methyl 3-O-benzyl-a-L-idopvranosvl-
uronateV(1?4)V3-O-benzyl-2-deoxy-2-r(benzyloxy^carbonvnamino-a-D-alucoDvranoside
(No. 101)
To a solution of compound 96 (570 mg, 194 umol) in methanol (66 mL) is added
ammonium fluoride (575 mg, 80 molar equivalents). After magnetic stirring at room
temperature for 14 hours, the reaction mixture is concentrated under vacuum and then
purified on a column of silica (toluene/acetone) to give compound 101 (282 mg, 54%).
Rf = 0.37 (7/3 toluene/acetone).
Preparation of methyl (methyl 3,4-di-O-benzyl-2-O-triethvlammonium sulfonato-
q-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-6-O-terf-butvldiphenvlsilvl-2-
deoxy-a-D-qlucopyranosyllMI —»4Hmethvl 3-O-benzyl-2-O-triethvlammonium sulfonato-
a-L-idopvranosvluronate)-(1^4V(2-azido-3-O-benzyl-2-deoxy-6-O-triethylammonium
sulfonato-g--D-qlucopyranosvIM 1-*4 Hmethvl_______3-O-benzyl-2-O-triethylammoniurn
sulfonato-a-L-idopvranosvluronate)-(1?4))-(2-azido-3-O-benzyl-2-deoxy-6-O-triethvl-
ammonium sulfonato-O--D-qlucopyranosv0-(1 -»4Hmethyl 3-O-benzyl-2-O-triethvl-
ammonium sulfonato-O-L-idopyranosvluronateM1?4)-3-O-benzyl-2-deoxy-2-r(benzyl-
oxy)carbonyllamino-6-O-triethylammonium sulfonato-a-D-glucopyranoside (No. 102)
Triethylamine/sulfur trioxide complex (376 mg, 5 mol per hydroxyl function) is
added to a solution in A/,A/-dimethylformamide (5.3 mL, 90 L/mol) of compound 101
(160 mg, 59 umol). After 17 hours of magnetic stirring at 55°C sheltered from light,
methanol is added at 0°C and, after stirring for 30 minutes at 0°C and then for
30 minutes at room temperature, the reaction medium is diluted with methanol and then
purified by means of an LH-20 column, using a 1/1 mixture of ethanol/dichloromethane
as eluent. The fractions containing the product are then concentrated under high
vacuum. The compound obtained is then used directly in the following step.
Rf = 0.52 (11/7/1.6/4 EtOAc/pyridine/AcOH/H20).
Preparation of methyl (methyl 2-O-ammonium sulfonato-3.4-di-O-benzyl-a-L-
idopvranosvluronate)-(1?4))-(2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosvl)-( 1?4)-
(methvl 2-O-ammonium sulfonato-3-O-benzyl-a-L-idopvranosyluronate)-(1?4)-(6-O-
ammonium sulfonato-2-azido-3-O-benzyl-2-deoxy-a-D-glucopyranosvl)-(1?4)-(methyl 2-
O-ammonium sulfonato-3-O-benzyl-a-L-idopyranosvluronate)-(1?4)-(6-O-ammonium
sulfonato-2-azido-3-O-benzyl-2-deoxy-O--D-qlucopyranosvl)-(1-»-4)-(methvl________2-O-
ammonium______sulfonato-S-O-benzyl-O-L-idopyranosvluronateVd—»4)-6-O-ammonium
sulfonato-3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonvnamino-o--D-qlucopyranoside (No.
103)
To a solution of compound 102 obtained previously in methanol (2.5 mL) is
added ammonium fluoride (42 mg, 60 molar equivalents). After magnetic stirring at
room temperature for 3 hours, at 45°C for 18 hours, and then at 55°C for 4 hours,
ammonium fluoride (14 mg, 20 molar equivalents) is again added, and the reaction
mixture is stirred for 24 hours at 55°C and then at room temperature for 60 hours. The
reaction mixture is then purified by means of an LH-20 column, using a 75/20/5 mixture
of methanol/N,N-dimethylformamide/water as eluent. The fractions containing the
product are then concentrated under high vacuum to give compound 103 (35 mg, 54%).
Rf = 0.54 (11/7/1.6/4 EtOAc/pyridine/AcOH/H20).
Preparation of methyl (lithium 3,4-di-O-benzyl-2-O-lithium sulfonato-O-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosvl)-(1?4)-
(lithium 3-O-benzyl-2-O-lithium sulfonato-a-L-idopyranosvluronate)-(1?4)-(2-azido-3-O-
benzyl-2-deoxy-6-O-lithium sulfonato-a-D-qlucopyranosyllM1?4Hlithium 3-O-benzyl-2-
O-lithium sulfonato-O-L-idopyranosvluronate)-(1?4M2-azido-3-O-benzyl-2-deoxy-6-O-
lithium sulfonato-a-D-qlucopyranosyllM1—»4Mlithium 3-O-benzyl-2-O-lithium sulfonato-
a-L-idopvranosvluronate)-(1?4)V3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonvnamino-6-O-
lithium sulfonato-g--D-qlucopyranoside (No. 104)
To a solution of compound 103 (155 mg, 41.6 umol) in a 1/1 mixture of
methanol/tetrahydrofuran (7 ml_) is added, dropwise and at 0°C, aqueous 1N LiOH
solution (1.4 mL, 8.4 equivalents/methyl ester function). After stirring for 1 hour at 0°C
and then for 15 hours at room temperature, the reaction medium is purified by means of
an LH-20 column, using a 95/5 methanol/water mixture as eluent. The fractions
containing the product are then concentrated under high vacuum to give the desired
compound 104 (114 mg, 90%).
Rf = 0.25 (11/7/1.6/4 EtOAc/pyridine/AcOH/H20).
Preparation of methyl (sodium 2-O-sodium sulfonato-O-L-idopyranosyluronate)-
(1?4H2-amino-2-deoxy-O--D-qlucopyranosvlH1?4>(sodium 2-O-sodium sulfonato-g-
L-idopvranosyluronate)-(1?4H2-amino-2-deoxy-6-O-sodium______sulfonato-O--D-gluco-
PVranosvl)-(1?4)-(sodium 2-O-sodium sulfonato-O-L-idopyranosvluronateH1?4)-(2-
amino-2-deoxy-6-O-sodium sulfonato-g--D-qlucopyranosvP-d—>4)-(sodium 2-O-sodium
sulfonato-g-L-idopvranosvluronateH1?4)-2-amino-2-deoxy-6-O-sodium sulfonato-O-p-
qlucopyranoside (No. 105)
To a solution in a 1/1 mixture of terf-butanol/water (8 mL) of compound 104
(110 mg, 36 umol) are successively added ammonium formate (298 mg, 4.72 mmol)
and then 10% Pd/C (715 mg). After 4 hours 15 minutes of vigorous stirring at room
temperature, the reaction medium is filtered and partially concentrated under vacuum,
and the solution is then applied to a Sephadex® G-25 column eluted with 0.2 M sodium
chloride. The fractions containing the product are concentrated and desalified using the
same column eluted with water. The fractions containing the product are then
concentrated under high vacuum to give the desired compound 105 (63 mg, 80%).
Chemical shifts of the anomeric protons (600 MHz, D20) 8 5.08 ldoUAvl", 5.23*
Glcv", 5.14 ldoUAvl, 5.24* Glcv, 5.14 ldoUAlv, 5.24* Glc'", 5.14 IdoUA", 4.73 Glc1
*: The signals may be interchanged.
Preparation of methyl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-
idopvranosvluronateV(1?4)V(2-azido-3-O-benzyl-2-deoxy-6-O-feff-butvldiphenvlsilvl--D-
D-alucopvranosvn-(1?4Hmethvl 2-O-acetyl-3-O-benzyl-g-L-idopvranosvluronate)-
(1 -»4V(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-alucopvranosvlV(1 -»4Mmethvl 2-O-
acetyl-3-O-benzyl-a-L-idopvranosvluronateV(1^4V3-O-benzyl-6-O-terf-butvldiphenvl-
silvl-2-deoxy-2-f(benzyloxy)carbonyllamino-a-D-qlucopyranoside(No. 106)
A solution of fert-butyldimethylsilyl triflate (1M, 0.2 mol per mole of imidate) in
dichloromethane is added, under argon and at -15°C, to a solution of the imidate 63
(0.59 g, 0.54 mmol) and of the glycosyl acceptor 92 (0.58 g, 0.36 mmol) in
dichloromethane (25 mL) in the presence of 4 A molecular sieves (0.4 g). After
45 minutes at -15°C (TLC), further imidate 63 (0.39 g, 0.35 mmol) is added, followed by
addition of fert-butyldimethylsilyl triflate in dichloromethane (1M, 1.1 mL). After 1 hour
10 minutes at -15°C (TLC), solid sodium hydrogen carbonate is added to neutralize.
After filtering, washing with aqueous 2% sodium hydrogen carbonate solution, drying
(Na2SO4) and evaporating to dryness, the residue is purified by means of an LH-20
column, using a 1/1 ethanol/dichloromethane mixture as eluent. The fractions containing
the product are then concentrated under high vacuum and the residue is then purified
on silica gel (ethyl acetate/toluene) to give 106 (0.69 g, 75%).
Rf = 0.16 (9/1 toluene/acetone).
Preparation of methyl (methyl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-
(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-fe/t-butvldiphenvlsilvl-a-D-qlucopyranosvl)-
(1?4)-(methvl 2-O-acetyl-3-O-benzyl-g-L-idopvranosvluronateH1?4)-(6-O-acetyl-2-
azido-3-O-benzyl-2-deoxy-g--D-glucopvranosvlH 1?4Mmethvl 2-O-acetyl-3-O-benzyl-g-
L-idopvranosvluronate)-(1^-4)-3-O-benzyl-6-O-fert-butvldiphenvlsilvl-2-deoxy-2-r(benzyl-
oxy)carbonvriamino-g--D-glucopvranoside (No. 107)
To a solution of compound 106 (1.41 g, 0.552 mmol) in a 1/2 mixture of
toluene/ethanol (110 mL) is added hydrazine acetate (0.254 g, 2.76 molar equivalents).
After 2 hours 40 minutes of magnetic stirring, the mixture is concentrated under vacuum
and then purified on silica gel to give compound 107 (1.15 g, 85%).
Rf = 0.16 (9/1 toluene/acetone).
Preparation of methyl (methyl 2-O-acetyl-3.4-di-O-benzyl-g-L-idopyranosvl-
uronate)-(1?4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-g--D-glucopyranosvn-(1?4)-
(methvl 2-O-acetyl-3-O-benzyl-g-L-idopvranosvluronate)-(1^-4)-(2-azido-3-O-benzyl-2-
deoxy-6-O-ferf-butvldiphenvlsilvl-g--D-glucopyranosvl)-(1^4Mmethvl 2-O-acetyl-3-O-
benzyl-g-L-idopvranosvluronate)-(1^4V(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-g--D-
glucopvranosv0-(1?4)-(methvl 2-O-acetyl-3-O-benzyl-g-L-idopvranosvluronate)-(1?4')-
3-O-benzyl-6-O-fert-butvldiphenvlsilvl-2-deoxy-2-r(benzyloxy)carbonvnamino-a-D-gluco-
pyranoside (No. 108)
A 0.1 M solution of tert-butyldimethylsilyl triflate in dichloromethane (1.1 mL) is
added, under argon and at -20°C, to a solution of the imidate 38 (0.67 g, 0.75 mmol)
and of the glycosyl acceptor 107 (1.23 g, 0.49 mmol) in dichloromethane (32 mL) in the
presence of 4 A molecular sieves (0.55 g). After 1.5 hours at -20°C (TLC), solid sodium
hydrogen carbonate is added to neutralize, and after filtering, washing with aqueous 2%
sodium hydrogen carbonate solution, drying (Na2SO4) and evaporating to dryness, the
residue is purified by means of an LH-20 column, using a 1/1 ethanol/dichloromethane
mixture as eluent. The fractions containing the product are then concentrated under
high vacuum and the residue is then purified on silica gel to give 108 (0.93 g, 59%).
Rf = 0.24 (7/3 cyclohexane/acetone).
Preparation of methyl (methyl 3,4-di-O-benzyl-a-L-idopvranosvluronateH 1?4V
(2-azido-3-O-benzyl-2-deoxy-a-D-glucopyranosvl)-(1?4)-(methvl_______3-O-benzyl-a-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-6-O-terf-butyldiphenvlsilyl-2-deoxy--D-
D-qlucopyranosvl)-(1?4)-(methvl 3-O-benzyl-O-L-idopvranosvluronateH1-»4M2-azido-
3-O-benzyl-2-deoxy-a-D-qlucopyranosylD-(1?4))-(methvl 3-O-benzyl-a-L-idopyranosvl-
uronate)-(1?4))-3-O-benzyl-6-O-fe/t-butvldiphenvlsilvl-2-deoxy-2-r(benzyloxy)carbonvn-
amino-g--D-qlucopyranoside (No. 109)
To a solution of 108 (1.17 g, 0.36 mmol) in a 3/2 mixture of
methanol/dichloromethane (109 mL) is added a 1M a methanolic solution of sodium
methoxide (1.6 mL), in the presence of 3 A molecular sieves (458 mg). After stirring for
18 hours at room temperature, the mixture is neutralized with H+ Dowex 50WX4 resin.
After filtering and concentrating, the residue is purified on silica gel with a
cyclohexane/ethyl acetate mixture to give compound 109 (680 mg, 69%).
Rf = 0.34 (7/3 toluene/acetone).
Preparation of methyl (methyl 3,4-di-O-benzyl-2-O-triethvlammonium sulfonato-
g-L-idopvranosyluronate)-(1—»4)-(2-azido-3-O-benzyl-2-deoxy-6-O-triethylammonium
sulfonato-o--D-glucopvranosvO-(1?4Hmethvl_______3-O-benzyl-2-O-triethvlammonium
sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-6-O-fe/t-butvldiphenyl-
silvl-g--D-glucopyranosvl)-(1?4)-(methvl 3-O-benzyl-2-O-triethylammonium sulfonato--D-
L-idopyranosvluronate)-(1—»4H2-azido-3-O-benzyl-2-deoxy-6-O-triethylammonium
sulfonato-a-D-glucopvranosvl)-(1?4)-(methyl_______3-O-benzyl-2-O-triethylammonium
sulfonato-g-L-idopvranosyluronate)-(1?4V3-O-benzyl-2-deoxy-2-r(berizvloxy)carbonvn-
amino-6-O-fert-butvldiphenvlsilvl-g--D-glucopyranoside (No. 110)
Triethylamine/sulfur trioxide complex (267 mg, 5 mol per hydroxyl function) is
added to a solution in A/,A/-dimethylformamide (4.4 mL, 90 L/mol) of compound 109
(144 mg, 49 umol). After 16 hours of magnetic stirring at 55°C sheltered from light,
methanol is added at 0°C and, after stirring for 30 minutes at 0°C and then for
30 minutes at room temperature, the reaction medium is diluted with methanol and then
purified by means of an LH-20 column, using a 1/1 mixture of
methanol/dichloromethane as eluent. The fractions containing the product are then
concentrated under high vacuum. The compound obtained is then used directly in the
following step.
Rf = 0.42 (57/29/7.2/16 EtOAc/pyridine/AcOH/H20).
Preparation of methyl (methyl 3.4-di-O-benzyl-2-O-ammonium sulfonato-a-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-ammonium sulfonato--D-
D-alucopvranosvlH1?4Mmethvl______3-O-benzyl-2-O-ammonium______sulfonato-a-L-
idopyranosyluronateV1?4)-(2-azido-3-O-benzyl-a-D-qlucopyranosyllH1?4)-fmethyl 3-
O-benzyl-2-O-ammonium_____sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-
benzyl-2-deoxy-6-O-ammonium sulfonato-a-D-qlucopyranosyll)-(1?4)-(methvl 3-O-
benzyl-2-O-ammonium sulfonato--D-i.-idopvranosvluronate)-(1?4)-3-O-benzyl-2-deoxy-
2-r(benzyloxy)carbonvnamino-a-D-qlucopyranoside (No. 111)
To a solution of compound 110 obtained previously in methanol (5 mL) is added
ammonium fluoride (133 mg, 80 molar equivalents). After magnetic stirring at 55°C for
48 hours, the reaction mixture is purified with the aid of an LH-20 column using a 95/5
methanol/water mixture as eluent. The fractions containing the product are then
concentrated under high vacuum. If necessary, the crude reaction product is used and
treated again under the same conditions to give compound 111 (117 mg, 85%, 2 steps).
Rf = 0.31 (57/29/7.2/16 EtOAc/pyridine/AcOH/H20).
Preparation of methyl (lithium 3,4-di-O-benzyl-2-O-lithium sulfonato-a-L-
idopvranosvluronate)-(1?4H2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-a-D-
qlucopyranosvlHI ->4)-(lithium 3-O-benzyl-2-O-lithium sulfonato-a-L-idopyranosyl-
uronate)-(1^-4)-(2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosyll)-(1?4)-(lithium 3-O-
benzyl-2-O-lithium sulfonato-a-L-idopvranosvluronate)-(1?4H2-azido-3-O-benzyl-2-
deoxy-6-O-lithium sulfonato-a-D-qlucopyranosylP-(1?4Hlithium 3-O-benzyl-2-O-lithium
sulfonato-a-L-idopvranosvluronate)-(1?4)-3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonyll-
amino-g--D-qlucopyranoside (No. 112)
To a solution of compound 111 (80 mg, 22.3 umol) in a 1/1 mixture of
methanol/tetrahydrofuran (3.6 mL) is added, dropwise and at 0°C, aqueous 1N LiOH
solution (0.7 mL, 7.8 equivalents/methyl ester function). After 2 hours 45 minutes of
stirring at 0°C, and then 17.5 hours at room temperature, the reaction medium is
purified by means of an LH-20 column, using a 75/20/5 mixture of methanol/A/./V-
dimethylformamide/water as eluent. The fractions containing the product are then
concentrated under high vacuum to give the desired compound 112 (64 mg, 98%).
Rf = 0.25 (56/32/7.6/18 EtOAc/pyridine/AcOH/H20).
Preparation of methyl (sodium 2-O-sodium sulfonato-a-L-idopvranosvluronate)-
(1?4H2-amino-2-deoxy-6-O-sodium sulfonato-a-D-a;lucopvranosvlH1?4Hsodium 2-
O-sodium_____sulfonato-a-L-idopvranosvluronateH1^4H2-amino-2-deoxy-a-D-gluco-
pyranosvlM1?4Msodium 2-O-sodium sulfonato-g-L-idopyranosvluronatey-d—»4H2-
amino-2-deoxy-6-O-sodium sulfonato-a-D-qlucopyranosylM1?4Msodium 2-O-sodium
sulfonato-a-L-idopvranosvluronate)-(1?4)V2-amino-2-deoxy-a-D-qlucopyranoside (No.
113)
To a solution in a 1/1 mixture of terf-butanol/water (4.3 ml_) of compound 112
(64 mg, 21.7 umol) are successively added ammonium formate (178 mg, 2.8 mmol) and
then 10% Pd/C (0.42 g). After 4 hours 30 minutes of vigorous stirring at room
temperature, the reaction medium is filtered and partially concentrated under vacuum,
and the solution is then applied to a Sephadex® G-25 column eluted with 0.2 M sodium
chloride. The fractions containing the product are concentrated and desalified using the
same column eluted with water. The fractions containing the product are then
concentrated under high vacuum to give the desired compound 113 (29.5 mg, 65%).
Chemical shifts of the anomeric protons (600 MHz, D20) 5 5.18 ldoUAvl", 5.43
Glcv", 5.24 ldoUAvl, 5.45 Glcv, 5.25 ldoUAlv, 5.43 Glc'", 5.26 IdoUA", 5.01 Glc1
Preparation of methyl (methyl 3,4-di-O-benzyl-a-L-idopyranosvluronate)-(1?4)-
(2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosyll)-(1^-4V(methvl_______3-O-benzyl-a-L-
idopyrariosvluronate)-(1^-4)-(2-azido-3-O-benzyl-6-O-fe/f-butvldiphenvlsilvl-2-deoxy--D-
D-qlucopyranosvl)-(1?4)-(methyl 3-O-benzyl-a-L-idopvranosvluronateM1—»4)-(2-azido-
3-O-benzyl-2-deoxy-g--D-olucopvranosvlH1?4Hmethvl 3-O-benzyl-a-L-idoDvranosvl-
uronateV(1^4V3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonvnamino-a-D-alucoDvranoside
(No. 114)
To a solution of compound 109 (135 mg, 46 umol) in methanol (16 ml_) is added
ammonium fluoride (136 mg, 80 molar equivalents). After magnetic stirring at room
temperature for 9 hours, and then at -25°C for 15 hours, the reaction mixture is
concentrated under vacuum and then purified on a column of silica (toluene/acetone) to
give compound 114 (57 mg, 46%).
Rf = 0.30 (toluene-methanol 85/15).
Preparation of methyl (methyl 3,4-di-O-benzyl-2-O-triethvlammonium sulfonato-
a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-triethylammonium
sulfonato-a-D-glucopvranosvlH1?4)-(methyl_______3-O-benzyl-2-O-triethvlammonium
sulfonato-a-L-idopyranosvluronate)-(1?4M2-azido-3-O-benzyl-6-O-ferf-butvldiphenvl-
silvl-2-deoxy-o--D-glucopvranosvO-(1?4)-(methyl 3-O-benzyl-2-O-triethvlammonium
sulfonato-g-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-triethyl-
ammonium sulfonato-g--D-glucopvranosvlH1 -»4)-(methyl 3-O-benzyl-2-O-triethyl-
ammonium sulfonato-g-L-idopvranosyluronateH1?4)-3-O-benzyl-2-deoxy-2-r(benzyl-
oxy)carbonvnamino-6-O-triethvlammoniumsulfonato-g--D-glucopyranoside (No. 115)
Triethylamine/sulfur trioxide complex (317 mg, 5 mol per hydroxyl function) is
added to a solution in A/,/V-dimethylformamide (4.5 mL, 90 L/mol) of compound 114
(135 mg, 50 umol). After 16 hours of magnetic stirring at 55°C sheltered from light,
methanol is added at 0°C and, after stirring for 30 minutes at 0°C and then for
30 minutes at room temperature, the reaction medium is diluted with methanol and then
purified by means of an LH-20 column, using a 1/1 mixture of ethanol/dichloromethane
as eluent. The fractions containing the product are then concentrated under high
vacuum. Compound 115 (168 mg, 86%) is then obtained.
Rf = 0.29 (56/32/7.6/18 EtOAc/pyridine/AcOH/H20).
Preparation of methyl (methyl 2-O-ammonium sulfonato-3,4-di-O-benzyl-o-L-
idopvranosvluronateM1?4H6-O-ammonium sulfonato-2-azido-3-O-benzyl-2-deoxy-g-
D-glucopyranosvD-(1?4Hmethvl______2-O-ammonium______sulfonato-3-O-benzyl-g-L-
idopvranosvluronate)-(1^-4V(2-azido-3-O-benzyl-2-deoxy-g--D-glucopvranosvl)-(1?4)-
(methyl 2-O-ammonium sulfonato-3-O-benzyl-a-L-idopyranosvluronate)-(1-»4V(6-O-
ammonium sulfonato-2-azido-3-O-benzyl-2-deoxy-a-D-glucopyranosvO-(1?4Hmethyl 2-
O-ammonium sulfonato-3-O-benzyl-g-L-idopvranosvluronateH1?4)-6-O-ammonium
sulfonato-3-O-benzyl-2-deoxy-2-[(benzyloxy)carbonvnamino-g--D-glucopyranoside (No.
116)
To a solution of compound 115 (168 mg, 43 umol) in methanol (5.6 ml_) is added
ammonium fluoride (128 mg, 80 molar equivalents). After magnetic stirring at 55°C for
48 hours, the reaction mixture is purified with the aid of an LH-20 column using a 95/5
methanol/water mixture as eluent. If necessary, the residue may be used under the
same conditions, while monitoring the reaction progress (TLC), and may be purified in
the same manner. The fractions containing the product are then concentrated under
high vacuum to give compound 116 (109 mg, 81%).
Rf = 0.21 (56/32/7.6/18 EtOAc/pyridine/AcOH/H20).
Preparation of methyl (lithium 3,4-di-O-benzyl-2-O-lithium sulfonato-a-L-
idopyranosvluronate)-(1?4M2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-a-D-
qlucopyranosyllM1?4)-(lithium 3-O-benzyl-2-O-lithium sulfonato-a-L-idopyranosyl-
uronateMI ->4)-(2-azido-3-O-benzyl-2-deoxy-a-D-qlucopvranosyl)-(1?4)-(lithium 3-O-
benzyl-2-O-lithium sulfonato--D-i_-idopvranosvluronate)-(1?4H2-azido-3-O-benzyl-2-
deoxy-6-O-lithium sulfonato-a-D-qlucopyranosyllH1?4Hlithium 3-O-benzyl-2-O-lithium
sulfonato-a-L-idopvranosvluronate)-(1?4))-3-O-benzyl-2-deoxy-2-f(benzyloxy)carbonvn-
amino-6-O-lithiumsulfonato-a-D-qlucopyranoside (No. 117)
To a solution of compound 116 (89 mg, 28.5 umol) in a 1/1 mixture of
methanol/tetrahydrofuran (3.8 mL) is added, dropwise and at 0°C, aqueous 1N LiOH
solution (0.7 mL). After stirring for 3 hours at 0CC, and then for 19 hours at room
temperature, the reaction medium is purified by means of an LH-20 column, using a
75/20/5 mixture of methanol/N, N-dimethylformamide/water as eluent. The fractions
containing the product are then concentrated under high vacuum and if necessary, the
reaction is repeated and then worked up in the same manner to give the desired
compound 117 (70 mg, 81%).
Rf = 0.12 (56/32/7.6/18 EtOAc/pyridine/AcOH/H20).
Preparation of methyl (sodium 2-O-sodium sulfonato-a-L-idopvranosvluronate)-
(1—*4H2-amino-2-deoxy-6-O-sodium sulfonato-a-D-qlucopyranosyllM1?4Hsodium 2-
O-sodium______sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-amino-2-deoxy-a-D-qluco-
PvranosvlH1?4)-(sodium 2-O-sodium sulfonato-a-L-idopvranosvluronateH1?4H2-
amino-2-deoxy-6-O-sodium sulfonato-a-D-qlucopyranosyll)-(1?4)-(sodium 2-O-sodium
sulfonato-a-L-idopvranosvluronateH1?4)-2-amino-2-deoxy-6-O-sodium sulfonato-a-D-
qlucopyranoside (No. 118)
To a solution in a 1/1 mixture of te/t-butanol/water (2.3 mL) of compound 117
(35 mg, 11.6 umol) are successively added ammonium formate (95 mg, 1.51 mmol) and
then 10% Pd/C (229 mg). After 4 hours 30 minutes of vigorous stirring at room
temperature, the reaction medium is filtered and partially concentrated under vacuum,
and the solution is then applied to a Sephadex® G-25 column eluted with 0.2 M sodium
chloride. The fractions containing the product are concentrated and desalified using the
same column eluted with water. The fractions containing the product are then
concentrated under high vacuum to give the desired compound 118 (11.1 mg, 45%).
Chemical shifts of the anomeric protons (600 MHz, D20) 8 5.17 ldoUAvl", 5.42
Glcv", 5.25 ldoUAvl, 5.45 Glcv, 5.24 ldoUAlv, 5.44 Glc'", 5.26 IdoUA", 5.03 Glc'
Preparation of 5-phenvlpentyl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-
idopvranosvluronate)-(1?4)-6-O-acetyl-3-O-benzyl-2-deoxy-2-f(benzyloxy)carbonvn-
amino-3--D-qlucopyranoside (No. 119)
ferf-Butyldimethylsilyl triflate (1.13 mL, 0.2 mol per mole of imidate) in
dichloromethane is added, under argon and at -20°C, to a solution of the imidate 24
(24.9 g, 24.6 mmol) and 5-phenylpentanol (8.3 mL, 2 molar equivalents) in
dichloromethane (1.1 L) in the presence of 4 A molecular sieves (31 g). After
10 minutes (TLC), saturated aqueous sodium hydrogen carbonate solution at 0°C is
added, and stirring is continued for 1 hour. The solution is filtered, washed with aqueous
2% sodium hydrogen carbonate solution, with saturated aqueous sodium chloride
solution, dried (Na2SO4) and evaporated to dryness. Purification on silica gel gives 119
(21.9 g, 88%).
LC-MS m/z 1034.3 [(M + Na)+]. TR = 1.80 min
Preparation of 2-propylpentyl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-
idopyranosvluronate)-(1->'4)-6-O-acetyl-3-O-benzyl-2-deoxy-2-r(benzyloxy')carbonyll-
amino-B--D-alucopvranoside (No. 120)
A 0.1 M solution of terf-butyldimethylsilyl triflate (2.78 mL, 0.2 mol per mole of
imidate) in dichloromethane is added, under argon, at -20°C, to a solution of the imidate
24 (1.2 g, 1.19 mmol) and 2-propylpentanol (1 mL, 5 molar equivalents) in
dichloromethane (63 mL) in the presence of 4 A molecular sieves (1.8 g). After
10 minutes (TLC), saturated aqueous sodium hydrogen carbonate solution is added at
0°C. The solution is filtered, washed with water, dried (Na2SO4) and evaporated to
dryness. Purification on silica gel gives 120 (0.91 g, 76%).
LC-MS m/z 1000.3 [(M + Na)*]. TR = 1.86 min
Preparation of 3-cyclohexylpropyl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-
a-L-idopvranosvluronate)-(1?4)-6-O-acetyl-3-O-benzyl-2-deoxy-2-r(benzyloxyV
carbonvnamino-B--D-qlucopvranoside (No. 121)
Compound 24 (1.2 g, 1.19 mmol) is treated according to a protocol similar to that
described for the synthesis of 120 to give compound 121 (1.09 g, 91 %).
LC-MS m/z 1012.3 [(M + Na)+]. TR = 1.88 min
Preparation of 3,3-diphenylpropvl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-
a-L-idopvranosvluronate)-(1?4)-6-O-acetyl-3-O-benzyl-2-deoxy-2-f(benzyloxy)-
carbonyllamino-g--D-qlucopyranoside (No. 122)
Compound 24 (1.2 g, 1.19 mmol) is treated according to a protocol similar to that
described for the synthesis of 120 to give compound 122 (1.02 g, 82%).
Rf = 0.34 (7/3 cyclohexane/acetone).
Preparation of 5-phenvlpentvl (methyl 2-O-acetyl-3-O-benzyl-4-O-levulinoyl--D-i_-
idopvranosvluronate)-(1^-4)-3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonyl1amino-8--D-
qlucopyranoside (No. 123)
To a solution of 119 (21.9 g, 21.6 mmol) in a 1/1 mixture of
methanol/tetrahydrofuran (260 mL) is added [tBu2SnCI(OH)]2 (0.926 g, 0.15 molar
equivalent) prepared according to A. Orita et al., Chem. Eur. J. (2001) 7, 3321. After
stirring at 35°C for 7.5 hours and then at room temperature for 17 hours, the reaction
mixture is concentrated under vacuum and then used directly in the following step.
LC-MS m/z 992.3 [(M + Na)+]. TR = 1.74 min
Preparation of 2-propylpentvl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-g-L-
idopvranosvluronate)-(1?4)-3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonvnamino-3--D-
glucopyranoside (No. 124)
To a solution of 120 (0.9 g, 0.93 mmol) in a 1/1 mixture of
methanol/tetrahydrofuran (12 mL) is added [tBu2SnCI(OH)]2 (0.037 g, 0.14 molar
equivalent) prepared according to A. Orita et al., Chem. Eur. J. (2001) 7, 3321. After
stirring at room temperature for 16 hours and then at 35°C for 4 hours, the reaction
mixture is concentrated under vacuum and then used directly in the following step.
LC-MS m/z 959.2 [(M + Na)+]. TR = 1.79 min
Preparation of 3-cvclohexvlpropvl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-
a-L-idopvranosvluronate)-(1?4))-3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonvnamino-3--D-
qlucopyranoside (No. 125)
Compound 121 (1.09 g, 1.10 mmol) is treated according to a protocol similar to
that described for the synthesis of 124 to give compound 125, which is used directly in
the following step.
LC-MS m/z 970.3 [(M + Na)+]. TR = 10.75 min
Preparation of 3,3-diphenylpropyl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-
a-L-idopvranosvluronate)-(1^4V3-O-benzyl-2-deoxy-2-[(benzyloxy)carbonyl1amino-3--D-
qlucopvranoside (No. 126)
Compound 122 (0.844 g, 0.80 mmol) is treated according to a protocol similar to
that described for the synthesis of 124. In the event of the reaction being incomplete,
the same amount of reagent may optionally be added again. After workup, compound
126 is used directly in the following step.
Rf = 0.41 (3/2 cyclohexane/acetone).
Preparation of 5-phenylpentyl (methyl 2-O-acetyl-3-O-benzyl-4-O-levulinoyl-a-L-
idopvranosvluronate)-(1?4))-3-O-benzyl-6-O-fe/,f-butvldiphenvlsilvl-2-deoxy-2-r(benzyl-
oxy)carbonvnamino-3--D-qlucopyranoside (No. 127)
Compound 123 obtained previously is dissolved in dichloromethane (173 mL).
Triethylamine (7.6 mL, 2.5 molar equivalents), 4-dimethylaminopyridine (1.3 g, 0.5
molar equivalent), and ferf-butyldiphenylsilyl chloride (11.2 mL, 2 molar equivalents) are
successively added at 0°C under argon. After 5.5 hours of magnetic stirring, the
reaction medium is diluted with dichloromethane, washed with aqueous 10% potassium
hydrogen sulfate solution, with saturated aqueous sodium chloride solution, dried
(Na2SO4), filtered and concentrated. The residue obtained is partially purified on silica
(acetone - toluene) which is used in the following step.
LC-MS m/z 1231.2 [(M + Na)+]. TR = 2.13 min
Preparation of 2-propylpentyl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl--D-i_-
idopvranosvluronateM1^4V3-O-benzyl-6-O-terf-butvldiphenvlsilvl-2-deoxy-2-r(benzyl-
oxy)carbonyllamino-S--D-qlucopyranoside (No. 128)
Compound 124 obtained previously is dissolved in dichloromethane (7.4 mL). 4-
Dimethylaminopyridine (57 mg, 0.5 molar equivalent), triethylamine (0.32 mL, 2.5 molar
equivalents), and fe/t-butyldiphenylsilyl chloride (0.48 mL, 2.0 molar equivalents) are
successively added at 0°C under argon. After 18 hours of magnetic stirring, the reaction
medium is diluted with dichloromethane, washed with aqueous 10% potassium
hydrogen sulfate solution, with aqueous 2% sodium hydrogen carbonate solution and
then with saturated aqueous sodium chloride solution, dried (Na2SO4), filtered and
concentrated. The residue obtained is purified on silica (acetone - cyclohexane) to give
128 (962 mg, 88% (2 steps)).
Rf = 0.46 (3/2 cyclohexane/acetone).
Preparation of 3-cvclohexvlpropyl (methyl 2-O-acetyl-3-O-benzyl-4-O-levulinoyl-
a-L-idopvranosvluronate)-(1^-4)-3-O-benzyl-6-O-teAt-butvldiphenylsilvl-2-deoxy-2-
r(benzyloxy)carbonvnamino-S--D-qlucopyranoside (No. 129)
Compound 125 obtained previously is treated according to a protocol similar to
that described for the synthesis of 128, to give compound 129 (1.12 g, 86% (2 steps)).
LC-MS m/z 1208.4 [(M + Na)+]. TR = 13.01 min
Preparation of 3.3-diphenylpropyl (methyl 2-O-acetyl-3-O-benzyl-4-O-levulinoyl-
a-L-idopvranosvluronate)-(1^-4)-3-O-benzyl-6-O-fe/t-butvldiphenvlsilvl-2-deoxy-2-
r(benzyloxy)carbonvnamino-B--D-qlucopyranoside (No. 130)
Compound 126 obtained previously is treated according to a protocol similar to
that described for the synthesis of 128, to give compound 130 (0.97 g, 81% (2 steps)).
LC-MS m/z 1278.3 [(M + Na)+]. TR = 12.72 min
Preparation of 5-phenvlpentvl (methyl 2-O-acetyl-3-O-benzyl-a-L-idopvranosvl-
uronate)-(1?4)-3-O-benzyl-6-O-te/t-butvldiphenvlsilvl-2-deoxy-2-r(benzyloxy)carbonyll-
amino-3--D-qlucopyranoside (No. 131)
To a solution of compound 127 obtained previously in a 1/2 mixture of
toluene/ethanol (1.01 L) is added hydrazine acetate (9.3 g, 5 molar equivalents). After
1 hour 10 minutes of magnetic stirring, the mixture is concentrated under vacuum and
then purified on silica gel to give compound 131 (16.7 g, 70% (3 steps)).
LC-MS m/z 1133.3 [(M + Na)+]. TR = 2.12 min
Chemical shifts of the anomeric protons (500 MHz, CDCI3) 8
5.19 IdoUA", 4.63 Glc'
Preparation of 2-propvlpentvl (methyl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvl-
uronate)-(1?4))-3-O-benzyl-6-O-terf-butvldiphenvlsilyl-2-deoxy-2-[(benzyloxy)carbonvn-
amino-B--D-qlucopvranoside (No. 132)
To a solution of compound 128 (0.96 g, 0.82 mmol) in a 1/2 mixture of
toluene/ethanol (164 ml_) is added hydrazine acetate (0.38 g, 5 molar equivalents). After
1 hour of magnetic stirring, the mixture is concentrated under vacuum and then purified
on silica gel to give compound 132 (0.84 g, 96%).
Rf = 0.42 (7/3 cyclohexane/acetone).
Mass: "ESI" method, positive mode: theoretical mass = 1076.3326; experimental
mass: 1098.5109 [M+Na]+.
Chemical shifts of the anomeric protons (500 MHz, CDCI3) 8
5.15 IdoUA", 4.61 Glc1
Preparation of 3-cvclohexvlpropyl (methyl 2-O-acetyl-3-O-benzyl-a-L-
idopvranosvluronate)-(1?4)-3-O-benzyl-6-O-fe/t-butvldiphenvlsilvl-2-deoxy-2-r(benzyl-
oxy)carbonyllamino-3--D-qlucopyranoside (No. 133)
Compound 129 (1.12 g, 0.94 mmol) is treated according to a protocol similar to
that described for the synthesis of 132, to give compound 133 (1.03 g, 98%).
LC-MS m/z 1110.4 [(M + Na)+]. TR = 13.05 min
Chemical shifts of the anomeric protons (500 MHz, CDCI3) 8
5.22 IdoUA", 4.64 Glc'
Preparation of 3,3-diphenvlpropvl (methyl 2-O-acetyl-3-O-benzyl-a-L-
idopvranosvluronate)-(1-»4V3-O-benzyl-6-O-teAt-butvldiphenvlsilvl-2-deoxy-2-r(benzyl-
oxy)carbonvnamino-B--D-qlucopyranoside (No. 134)
Compound 130 (0.96 g, 0.77 mmol) is treated according to a protocol similar to
that described for the synthesis of 132, to give compound 134 (833 mg, 94%).
LC-MS m/z 1180.5 [(M + Na)+]. TR = 12.22 min
Chemical shifts of the anomeric protons (500 MHz, CDCI3) 5
5.21 IdoUA", 4.54 Glc'
Preparation of 5-phenylpentyl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-
idopyranosvluronate)-(1—»4)-(2-azido-3-O-benzyl-6-O-fe/t-butvldiphenvlsilyl-2-deoxy--D-
D-glucopyranosyl)-(1?4)-(methvl_____2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-
(1?4)-3-O-benzyl-6-O-fe/t-butvldiphenvlsilvl-2-deoxy-2-f(benzyloxy)carbonyl1amino-B--D-
qlucopyranoside (No. 135)
te/t-Butyldimethylsilyl triflate (0.51 mL, 0.15 mol per mole of imidate) in
dichloromethane is added, under argon and at -20°C, to a solution of the imidate 63
(24.7 g, 22.5 mmol) and of the glycosyl acceptor 131 (16.7 g, 15 mmol) in
dichloromethane (788 mL) in the presence of 4 A molecular sieves (16.9 g). After
2 hours 45 minutes at -20°C (TLC), solid sodium hydrogen carbonate is added. After
filtering, washing with aqueous 2% sodium hydrogen carbonate solution and with
saturated aqueous sodium chloride solution, followed by drying (Na2SO4) and
evaporating to dryness, the residue is purified on silica gel to give 135 (20.6 g, 67%).
Chemical shifts of the anomeric protons (500 MHz, CDCI3) 8 5.35 ldoUAlv, 4.77
Glc'", 5.30 IdoUA", 4.61 Glc'
Rf = 0.41 (9/1 toluene/acetone).
Preparation of 2-propylpentvl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-6-O-te/t-butvldiphenylsilvl-2-deoxy--D-
D-qlucopyranosyll)-(1?4Mmethyl_____2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-
(1?4)-3-O-benzyl-6-O-terf-butvldiphenvlsilvl-2-deoxy-2-r(benzyloxy)carbonvnamino-B--D-
qlucopyranoside (No. 136)
A 0.1 M solution of ferf-butyldimethylsilyl triflate in dichloromethane (1.1 mL,
0.15 mol per mole of imidate) is added, under argon and at -20°C, to a solution of the
imidate 63 (0.81 g, 0.73 mmol) and of the glycosyl acceptor 132 (0.84 g, 0.78 mmol) in
dichloromethane (26 mL) in the presence of 4 A molecular sieves (0.55 g). After 2 hours
30 minutes at -20°C (TLC), solid sodium hydrogen carbonate is added. After filtering,
washing with aqueous 2% sodium hydrogen carbonate solution and with saturated
aqueous sodium chloride solution, followed by drying (Na2SO4) and evaporating to
dryness, the residue is purified on silica gel to give 136 (0.66 g, 47%).
Rf = 0.23 (4/1 cyclohexane/acetone).
Preparation of 3-cyclohexvlpropyl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-
a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-6-O-te/t-butvldiphenvlsilvl-2-
deoxy-g--D-qlucopyranosylD-(1?4Hmethvl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvl-
uronate)-(1?4)-3-O-benzyl-6-O-ter(-butvldiphenvlsilvl-2-deoxy-2-[(benzyloxy)carbonvn-
amino-3--D-qlucopyranoside (No. 137)
The glycosyl donor 63 (0.63 g, 0.57 mmol) and the glycosyl acceptor 133
(1.03 g, 0.95 mmol) are treated according to a protocol similar to that described for the
synthesis of 136 to give compound 137 (716 mg, 62%).
Rf = 0.20 (3/1 cyclohexane/acetone).
Preparation of 3,3-diphenylpropyl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-
g-L-idopvranosyluronate)-(1—»4)-(2-azido-3-O-benzyl-6-O-fe/t-butvldiphenvlsilyl-2-
deoxy-a-D-qlucopyranosylP-(1—»4Mmethyl_____2-O-acetyl-3-O-benzyl-a-L-idopvranosvl-
uronate)-(1?4)-3-O-benzyl-6-O-fe/f-butvldiphenvlsilvl-2-deoxy-2-f(benzyloxy)carbonyll-
amino-3--D-glucopyranoside (No. 138)
The glycosyl donor 63 (0.63 g, 0.57 mmol) and the glycosyl acceptor 134
(0.83 g, 0.72 mmol) are treated according to a protocol similar to that described for the
synthesis of 136, to give compound 138.
Rf = 0.34 (7/3 cyclohexane/acetone).
Preparation of 5-phenylpentyl (methyl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvl-
uronate)-(1?4)-(2-azido-3-O-benzyl-6-O-te/t-butvldiphenvlsilyl-2-deoxy-a-D-qluco-
pyranosvlMI ->4)-(methvl 2-O-acetyl-3-O-benzyl-g-L-idopyranosvluronate)-(1?4)-3-O-
benzyl-6-O-tert-butvldiphenvlsilyl-2-deoxy-2-r(benzyloxy)carbonvnamino-6--D-gluco-
pyranoside (No. 139)
To a solution of compound 135 (20.6 g, 10.0 mmol) in a 1/2 mixture of
toluene/ethanol (500 mL) is added hydrazine acetate (4.6 g, 5 molar equivalents). After
1 hour 20 minutes of magnetic stirring, the mixture is concentrated under vacuum and
then purified on silica gel to give compound 139 (16.8 g, 86%).
Rf = 0.37 (9/1 toluene/acetone).
Preparation of 2-propvlpentvl (methyl 2-O-acetyl-3-O-benzyl-O-i_-idopyranosvl-
uronate)-(1?4)-(2-azido-3-O-benzyl-6-O-fe/t-butvldiphenvlsilyl-2-deoxy-g--D-qluco-
pyranosvlM1?4)-(methyl 2-O-acetyl-3-O-benzyl-g-L-idopyranosvluronateH1?4)-3-O-
benzyl-6-O-terf-butvldiphenvlsilvl-2-deoxy-2-f(benzyloxy)carbonvnamino-3--D-gluco-
pyranoside (No. 140)
To a solution of compound 136 (20.6 g, 10.0 mmol) in a 1/2 mixture of
toluene/ethanol (500 mL) is added hydrazine acetate (4.6 g, 5 molar equivalents). After
1 hour 20 minutes of magnetic stirring, the mixture is concentrated under vacuum and
then purified on silica gel to give compound 140 (16.8 g, 86%).
Rf = 0.36 (95/5 toluene/methanol).
Preparation of 3-cyclohexylpropvl (methyl 2-O-acetyl-3-O-benzyl-a-L-
idopyrariosvluronate)-(1?4)-(2-azido-3-O-benzyl-6-O-fe/t-butvlcliphenvlsilvl-2-deoxy--D-
D-qlucopyranosvl)-(1?4)-(methyl_____2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-
(1?4)-3-O-benzyl-6-O-tert-butvldiphenvlsilvl-2-deoxy-2-r(benzyloxy)carbonvnamino-3--D-
qlucopyranoside (No. 141)
Compound 137 (0.71 g, 0.35 mmol) is treated according to a protocol similar to
that described for the synthesis of 140, to give compound 141 (0.65 g, 96%).
Rf = 0.37 (7/3 cyclohexane/acetone).
Preparation of 3,3-diphenylpropyl (methyl 2-O-acetyl-3-O-benzyl-a-L-
idopvranosvluronate)-(1?4))-(2-azido-3-O-benzyl-6-O-ferf-butvldiphenvlsilyl-2-deoxy--D-
D-qlucopyranosyllH1?4Mmethvl_____2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-
(1?4)-3-O-benzyl-6-O-teAf-butvldiphenvlsilvl-2-deoxy-2-r(benzyloxy)carbonvnamino-B--D-
glucopyranoside (No. 142)
Compound 138 obtained in the preceding step is treated according to a protocol
similar to that described for the synthesis of 140, to give compound 142 (0.518 g, 45%
(2 steps)).
Rf = 0.37 (9/1 toluene/acetone).
Preparation of 5-phenylpentvl (methyl 2-O-acetyl-3-O-benzyl-4-O-leyulinoyl-a-L-
idopyranosvluronate)-(1?4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-gluco-
pyranosyD-1?4Hmethyl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-(1?4V(2-
azido-3-O-benzyl-6-O-teAf-butvldipherivlsilvl-2-deoxy-a-D-qlucopyranosyln-(1?4)-(methyl
2-O-acetyl-3-O-benzyl-a-L-idopvranosvluronate)-(1^-4)-3-O-benzyl-6-O-fe/t-butyl-
diphenvlsilvl-2-deoxy-2-r(benzyloxy)carbonvnamino-S--D-qlucopyranoside (No. 143)
tert-Butyldimethylsilyl triflate (0.42 mL, 0.15 mol per mole of imidate) in
dichloromethane is added, under argon and at -20°C, to a solution of the imidate 27
(11.1 g, 12.3 mmol) and of the glycosyl acceptor 139 (16.8 g, 8.6 mmol) in
dichloromethane (430 mL) in the presence of 4 A molecular sieves (10.4 g). After
3 hours of magnetic stirring at -20°C (TLC), solid sodium hydrogen carbonate is added.
After filtering, washing with aqueous 2% sodium hydrogen carbonate solution and with
saturated aqueous sodium chloride solution, followed by drying (Na2SO4) and
evaporating to dryness, the residue is purified on silica gel to give 143 (14.71 g, 64%).
Rf = 0.31 (85/15 toluene/acetone).
Preparation of 5-phenvlpentvl (methyl 2-O-acetyl-3-O-benzyl-a-L-idopvranosyl-
uronateH1^4H6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosyllH1?4)-
(methvl 2-O-acetyl-3-O-benzyl-a-L-idopvranosvluronate)-(1-»4V(2-azido-3-O-benzyl-6-
O-ferf-butvldiphenvlsilvl-2-deoxy-a-D-q)ucopvranosv0-(1?4Hmethvl 2-O-acetyl-3-O-
benzyl-a-L-idopvranosvluronateH1-»4)-3-O-benzyl-6-O-fe/f-butvldiphenvlsilvl-2-deoxy-
2-[(benzyloxy)carbonvnamino-3--D-glucopvranoside (No. 144)
To a solution of compound 143 (13.51 g, 5.0 mmol) in a 1/2 mixture of
toluene/ethanol (252 ml_) is added hydrazine hydrate (2.31 g, 5 molar equivalents).
After 30 minutes of magnetic stirring, the mixture is concentrated under vacuum and
then purified on silica gel to give compound 144 (9.41 g, 72%).
Rf = 0.18 (85/15 toluene/acetone).
Preparation of 5-phenvlpentvl (methyl 2-O-acetyl-3.4-di-O-benzyl--D-i_-
idopvranosvluronateVd—>4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qluco-
PVranosvlH1?4)-(methvl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-(1?4)-(6-O-
acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosyll)-(1—»4)-(methvl 2-O-acetyl-3-O-
benzyl-a-L-idopvranosvluronateH1-»4)-(2-azido-3-O-benzyl-6-O-te/t-butvldiphenylsilvl-
2-deoxy-a-D-qlucopyranosylD-(1?4)-(methvl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvl-
uronate)-(1^-4)-3-O-benzyl-6-O-ferf-butvldiphenvlsilvl-2-deoxy-2-f(benzyloxy)carbonvn-
amino-B--D-glucopvranoside (No. 145)
A 0.1 M solution of terf-butyldimethylsilyl triflate (7.2 ml_, 0.15 mol per mole of
imidate) in dichloromethane is added, under argon, at -20°C, to a solution of the imidate
38 (4.29 g, 4.8 mmol) and of the glycosyl acceptor 144 (10.33 g, 4.0 mmol) in
dichloromethane (170 mL) in the presence of 4 A molecular sieves (3.7 g). After
19 hours 45 minutes at -20°C (TLC), solid sodium hydrogen carbonate is added. After
filtering, washing with aqueous 2% sodium hydrogen carbonate solution, with saturated
aqueous sodium chloride solution, followed by drying (Na2SO4) and evaporating to
dryness, the residue is purified on silica gel to give 145 (7.74 g, 58%).
Rf = 0.4 (85/15 toluene/acetone).
Preparation of (methyl 2-O-acetyl-3-O-benzyl-a-L-idopvranosvluronateH1?4)-
1,6-anhvdro-2-azido-3-O-benzyl-2-deoxy-3--D-qlucopyranose (No. 147)
To a solution of compound 146 (11.1 mmol) (WO 2006/021 653) in N,N-
dimethylformamide (78 mL) is added, at 0°C and under argon, potassium hydrogen
carbonate (5.56 g, 5 molar equivalents), followed by addition of methyl iodide (6.9 mL,
10 molar equivalents). After 17 hours of magnetic stirring, the reagents may be added
once again, if necessary. Once the reaction is complete (TLC), the reaction medium is
concentrated under vacuum, the crude reaction product is diluted with ethyl acetate,
washed with water and then with saturated aqueous sodium thiosulfate solution, dried
(Na2SO4), filtered and concentrated. The residue obtained is used in the following step
without purification.
LC-MS m/z 465.3 [(M + NH4)+]. TR = 0.8 min
Preparation of (methyl 2-O-acetyl-3,4-di-O-benzyl-a-L-idopvranosvluronate)-
(1?4H6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosyllH1?4)-(methyl 2-
0-acetyl-3-O-benzyl-a-L-idopvranosvluronate)-(1?4)-1,6-anhvdro-2-azido-3-O-benzyl-
2-deoxy-3--D-qlucopyranose (No. 148)
A 1 M solution of terf-butyldimethylsilyl triflate (0.95 mL, 0.15 mol per mole of
imidate) in dichloromethane is added, under argon and at -20°C, to a solution of the
imidate 38 (5.68 g, 6.35 mmol) and of the glycosyl acceptor 147 (4.2 g, 7 mmol) in
dichloromethane (222 mL) in the presence of 4 A molecular sieves (4.8 g). After
1.5 hours at -20°C (TLC), catalyst is optionally added until the reaction is complete
(TLC), and then solid sodium hydrogen carbonate is added. After filtering, washing with
aqueous 2% sodium hydrogen carbonate solution and with saturated sodium chloride
solution, drying (Na2SO4), filtering and evaporating to dryness, the residue is purified on
silica gel to give 148 (3.15 g, 38%).
Rf = 0.41 (3/2 cyclohexane/ethyl acetate).
Preparation of (methyl 2-O-acetyl-3,4-di-O-benzyl-a-L-idopvranosvluronate)-
(1?4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-glucopvranosyl)-(1?4Hmethyl 2-
0-acetyl-3-O-benzyl-a-L-idopvranosvluronate)-(1?4)-1,6-di-O-acetyl-2-azido-3-O-
benzyl-2-deoxy-a,B--D-qlucopvranose (No. 149)
The residue obtained in the preceding step is dissolved in acetic anhydride
(13 ml_), followed by addition, at 0°C over 15 minutes, of trifluoroacetic acid (TFA)
(1.3 ml_). The reaction mixture is stirred for 10 minutes at 0CC and for 18 hours at room
temperature, and is then concentrated, co-evaporated with toluene, and purified on
silica gel (heptane/ethyl acetate), to give compound 149 (2.99 g, 88%).
LC-MS m/z 1456.0 [(M + Na)+]. TR = 1.96 min
Preparation of (methyl 2-O-acetyl-3,4-di-O-benzyl-a-L-idopyranosvluronate)-
f1^-4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosyll)-(1?4)-(methvl 2-
0-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-(1?4)-6-O-acetyl-2-azido-3-O-benzyl-2-
deoxy-a,B--D-qlucopvranose (No. 150)
To a solution of compound 149 (2.99 g, 2.09 mmol) in diethyl ether (42 mL) is
added, at 0°C, benzylamine (BnNH2) (8.2 mL, 36 molar equivalents). After 1.5 hours of
stirring at 0°C, and then 4.5 hours at room temperature, the reaction mixture is
neutralized with cold (0-4°C) 1N HCI, washed with water, dried (Na2SO4), filtered and
concentrated, and purified on silica gel (heptane/ethyl acetate) to give 150 (2.6 g, 90%).
Rf = 0.21 (3/2 cyclohexane/ethyl acetate).
Preparation of (methyl 2-O-acetyl-3.4-di-O-benzyl-a-L-idopyranosvluronate)-
(1?4H6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qlucopvranosyl)-(1?4Hmethyl 2-O-
acetyl-3-O-benzyl-a-L-idopvranosvluronate)-(1?4)-6-O-acetyl-2-azido-3-O-benzyl-2-
deoxy-a,B--D-qlucopvranose trichloroacetimidate (No. 151)
To a solution of compound 150 (2.6 g, 1.9 mmol) in dichloromethane (36 mL) is
added, under argon and at 0°C, caesium carbonate (Cs2C03) (0.97 g, 1.6 molar
equivalents), followed by addition of trichloroacetonitrile (CCI3CN) (0.94 mL, 5.0 molar
equivalents). After stirring for 1 hour at room temperature, the reaction mixture is filtered
and then concentrated. The residue is purified on silica gel (ethyl acetate/cyclohexane +
0.1% triethylamine) to give 151 (2.25 g, 80%).
Preparation of 2-propylpentvl (methyl 2-O-acetyl-3,4-di-O-benzyl-g-i_-
idopvranosvluronate)-(1?4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qluco-
pvranosvlH1?4)-(methvl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvluronate)-(1?4)-(6-O-
acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-glucopyranosvl)-(1?4)-(methvl 2-O-acetyl-3-O-
benzyl-a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-6-O-te/t-butvldiphenvlsilvl-
2-deoxy-g--D-glucopvranosvlH1?4Hmethvl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvl-
uronate)-(1?4))-3-O-benzyl-6-O-ferf-butvldiphenvlsilvl-2-deoxy-2-f(benzyloxy)carbonyll-
amino-3--D-glucopyranoside (No. 152)
A 0.1 M solution of terf-butyldimethylsilyl triflate (0.46 mL, 0.15 mol per mole of
imidate) in dichloromethane is added, under argon and at -20°C, to a solution of the
imidate 151 (0.56 g, 0.36 mmol) and of the glycosyl acceptor 140 (0.59 g, 0.31 mmol) in
dichloromethane (11 mL) in the presence of 4 A molecular sieves (0.27 g). After 5 hours
at -20°C (TLC), solid sodium hydrogen carbonate is added. After filtering, washing with
aqueous 2% sodium hydrogen carbonate solution and with saturated aqueous sodium
chloride solution, followed by drying (Na2SO4) and evaporating to dryness, the residue is
purified on silica gel to give 152 (0.82 g, 82%).
Rf = 0.32 (7/3 cyclohexane/acetone).
Preparation of 3-cvclohexvlpropyl (methyl 2-O-acetyl-3,4-di-O-benzyl-o-L-
idopvranosvluronate)-(1?4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-gluco-
pvranosvlH1?4)-(methvl 2-O-acetyl-3-O-benzyl-a-L-idopvranosvluronateM1?4)-(6-O-
acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosvl)-(1?4)-(methvl 2-O-acetyl-3-O-
benzyl-g-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-6-O-ferf-butvldiphenvlsilvl-
2-deoxy-a-D-qlucopyranosyllH1?4)-(methvl 2-O-acetyl-3-O-benzyl-a-L-idopyranosvl-
uronate)-(1^-4)-3-O-benzyl-6-O-feAf-butvldiphenvlsilvl-2-deoxy-2-r(benzyloxy)carbonvn-
amino-g--D-qlucopyranoside (No. 153)
The glycosyl donor 151 (0.56 g, 0.36 mmol) and the glycosyl acceptor 141
(0.65 g, 0.34 mmol) are treated according to a protocol similar to that described for the
synthesis of 152, to give compound 153 (808 mg, 73%).
Rf = 0.36 (3/2 cyclohexane/acetone).
Preparation of 3,3-diphenvlpropvl (methyl 2-O-acetyl-3,4-di-O-benzyl-a-L-
idopvranosvluronate)-(1?4)-(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a-D-gluco-
pvranosv0-(1?4)-(methvl 2-O-acetyl-3-O-benzyl4)-(methvl 3-O-benzyl-a-L-idopvranosvluronate)-(1 ->-4)-(2-azido-3-O-
benzyl-6-O-ferf-butvldiphenvlsilvl-2-deoxy-a-D-qlucopyranosvn-(1?4)-(methvl_____3-O-
benzyl-a-L-idopvranosvluronate)-(1?4)-3-O-benzyl-6-O-te/t-butyldiphenvlsilvl-2-deoxy-
2-f(benzyloxy)carbonvnamino-S--D-qlucopyranoside (No. 157)
Compound 153 (402 mg, 0.122 mmol) is treated according to a protocol similar
to that described for the synthesis of 155, to give compound 157 (276 mg, 75%).
Rf = 0.40 (7/3 cyclohexane/acetone).
Preparation of 3,3-diphenylpropyl (methyl 3,4-di-O-benzyl-a-L-idopvranosvl-
uronateH1^4H2-azido-3-O-benzyl-2-deoxy-O--D-qlucopyranosyllH1-+4)-(methvl 3-O-
benzyl-a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-a-D-qluco-
pyranosylVd—»4)-(methyl 3-O-benzyl-a-L-idopyranosvluronate)-(1?4)-(2-azido-3-O-
benzyl-6-O-terf-butvldiphenvlsilyl-2-deoxy-O--D-qlucopyranosyllM1^4)-(methvl_____3-O-
benzyl-O-L-idopvranosvluronate)-(1?4)-3-O-benzyl-6-O-ferf-butvldiphenvlsilvl-2-deoxy-
2-[(benzyloxy)carbonvnamino-B--D-qlucopyranoside (No. 158)
Compound 154 (315 mg, 0.093 mmol) is treated according to a protocol similar
to that described for the synthesis of 155, to give compound 158 (244 mg, 84%).
Rf = 0.13 (7/3 cyclohexane/acetone).
Preparation of 5-phenvlpentyl (methyl 3,4-di-O-benzyl-2-O-triethvlammonium
sulfonato-a-L-idopvranosvluronate)-(1?4))-(2-azido-3-O-benzyl-2-deoxy-6-O-triethyl-
ammonium sulfonato-O--D-qlucopyranosyll)-(1?4)-(methvl 3-O-benzyl-2-O-triethvl-
ammonium sulfonato-o-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-
O-triethylammonium sulfonato-O--D-qlucopyranosyll)-(1?4)-(methvl 3-O-benzyl-2-O-
triethylammonium sulfonato-O-L-idopvranosvluronateH1?4H2-azido-3-O-benzyl-6-O-
tert-butvldiphenvlsilvl-2-deoxy-O--D-qlucopyranosvn-(1?4)-(methvl_____3-O-benzyl-2-O-
triethylammonium______sulfonato-O-i_-idopvranosvluronateM1?4y3-O-benzyl-6-O-ferf-
butvldiphenvlsilvl-2-deoxy-2-f(benzyloxy)carbonyllamino-B--D-qlucopyranoside (No. 159)
Triethylamine/sulfur trioxide complex (775 mg, 5 mol per hydroxyl function) is
added to a solution in A/,/V-dimethylformamide (12.8 mL, 90 L/mol) of compound 155
(438 mg, 142 umol). After 17 hours of magnetic stirring at 50°C sheltered from light,
methanol is added at 0°C and, after stirring for 30 minutes at 0°C and then 1 hour at
room temperature, the reaction medium is diluted with methanol and then purified by
means of an LH-20 column, using a 9/1 mixture of methanol/N,N-dimethylformamide as
eluent. The fractions containing the product are then concentrated under high vacuum
to give the desired compound 159 (571 mg, 96%).
Rf = 0.48 (29/13/3.4/7 EtOAc/pyridine/AcOH/H20).
Preparation of 2-propylpentyl (methyl 3,4-di-O-benzyl-2-O-triethvlammonium
sulfonato-O-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-triethvl-
ammonium sulfonato-a-D-qlucopyranosyll)-(1?4)-(methvl 3-O-benzyl-2-O-triethyl-
ammonium sulfonato-O-L-idopvranosvluronateH1?4)-(2-azido-3-O-benzyl-2-deoxy-6-
O-triethylammonium sulfonato-O--D-qlucopyranosvn-(1?4)-(methvl 3-O-benzyl-2-O-
triethylammonium sulfonato-O-L-idopvranosvluronate)-(1?4M2-azido-3-O-benzyl-6-O-
ferf-butvldiphenvlsilvl-2-deoxy-O--D-qlucopyranosvn-(1^-4)-(methvl_____3-O-benzyl-2-O-
triethylammonium sulfonato-a-L-idopyranosvluronate)-(1^-4)-3-O-benzyl-6-O-fe/t-butvl-
diphenvlsilvl-2-deoxy-24(benzyloxy)carbonvnamino-B--D-qlucopvranoside (No. 160)
Compound 156 (178 mg, 0.054 mmol) is treated according to a protocol similar
to that described for the synthesis of 159, to give compound 160 (196 mg, 75%).
Rf = 0.48 (57/29/7.2/16 EtOAc/pyridine/AcOH/H20).
Preparation of 3-cvclohexylpropvl (methyl 3,4-di-O-benzyl-2-O-triethylammonium
sulfonato-a-L-idopvranosvluronate)-(1^-4)-(2-azido-3-O-benzyl-2-deoxy-6-O-triethvl-
ammonium sulfonato-a-D-glucopyranosvl)-(1-+4Hmethvl 3-O-benzyl-2-O-triethvl-
ammonium sulfonato-O-L-idopvranosvluronate)-(1-*4H2-azido-3-O-benzyl-2-deoxy-6~
O-triethylammonium sulfonato-O--D-qlucopyranosyll)-(1?4)-(methvl 3-O-benzyl-2-O-
triethylammonium sulfonato-a-L-idopvranosvluronate)-(1?4))-(2-azido-3-O-benzyl-6-O-
te/t-butvldiphenvlsilvl-2-deoxy-a-D-qlucopyranosvl)-(1?4)-(methvl_____3-O-benzyl-2-0~
triethylammonium______sulfonato--D-i_-idopvranosvluronate)-(1?4)-3-O-benzyl-6-O-terf-
butvldiphenvlsilvl-2-deoxy-2-[(benzyloxy)carbonvnamino-3--D-qlucopyranoside (No. 161)
Compound 157 (345 mg, 0.113 mmol) is treated according to a protocol similar
to that described for the synthesis of 159, to give compound 161 (332 mg, 70%).
Rf = 0.48 (57/29/7.2/16 EtOAc/pyridine/AcOH/H20).
Preparation of 3,3-diphenvlpropvl (methyl 3,4-di-O-benzyl-2-O-triethvlammonium
sulfonato-O-L-idopvranosvluronate)-(1^-4)-(2-azido-3-O-benzyl-2-deoxy-6-O-triethvl-
ammonium sulfonato-a-D-qlucopvranosyl)-(1?4)-(methvl 3-O-benzyl-2-O-triethvl-
ammonium sulfonato--D-i_-idopvranosvluronate)-(1?4H2-azido-3-O-benzyl-2-deoxy-6-
O-triethylammonium sulfonato-O--D-qlucopyranosyll)-(1?4)-(methvl 3-O-benzyl-2-O-
triethvlammonium sulfonato-O-L-idopyranosvluronate)-(1?4))-(2-azido-3-O-benzyl-6-O-
ferf-butvldiphenvlsilvl-2-deoxy-a-D-qlucopyranosvl)-(1—»4)-(methyl_____3-O-benzyl-2-O-
triethylammonium______sulfonato-O-i_-idopvranosyluronate)-(1?4)-3-O-benzyl-6-O-ferf-
butvldiphenvlsilvl-2-deoxy-2-f(benzyloxy)carbonyl1amino-3-P-qlucopyranoside (No. 162)
Compound 158 (300 mg, 0.096 mmol) is treated according to a protocol similar
to that described for the synthesis of 159. If necessary, the mixture obtained may be
purified on an RP-18 reverse-phase column using a methanol/water mixture as eluent,
to give compound 162 (211 mg, 52%).
Rf = 0.47 (29/13/3.4/7 EtOAc/pyridine/AcOH/H20).
Preparation of 5-phenvlpentyl (methyl 3,4-di-O-benzyl-2-O-ammonium sulfonato-
a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-ammonium
sulfonato-a-D-qlucopyranosylO-(1—»4)-(methyl 3-O-benzyl-2-O-ammonium sulfonato-a-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-ammonium sulfonato-O-
D-qlucopyranosvD-d -»4Hmethvl______3-O-benzyl-2-O-ammonium______sulfonato-a-L-
idopvranosvluronateH1-^4M2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosvlH1?4)-
(methyl 3-O-benzyl-2-O-ammonium sulfonato-a-L-idopyranosvluronate)-(1?4)-3-O-
benzyl-2-deoxy-2-[(benzyloxy)carbonvnamino-B--D-qlucopyranoside (No. 163)
To a solution of compound 159 (520 mg, 125 umol) in methanol (16 mL) is
added ammonium fluoride (370 mg, 80 molar equivalents). After magnetic stirring at
55°C for 24 hours and then at room temperature for 16 hours, the reaction mixture is
purified with the aid of an LH-20 column using a 9/1 mixture of methanol/N,/V-
dimethylformamide as eluent. The fractions containing the product are then
concentrated under high vacuum and the residue is purified on silica gel with an ethyl
acetate/pyridine/acetic acid/water mixture as eluent. The fractions containing the
product are then partially concentrated and the residue is purified by means of an LH-20
column, using a 9/1 mixture of methanol/W,N-dimethylformamide as eluent. The
fractions containing the product are then concentrated under high vacuum to give
compound 163 (295 mg, 74%).
Rf = 0.45 (29/13/3.4/7 EtOAc/pyridine/AcOH/H20).
Preparation of 2-propylpentvl (methyl 3.4-di-O-benzyl-2-O-ammonium sulfonato-
a-L-idopvranosvluronateH1^4M2-azido-3-O-benzyl-2-deoxy-6-O-ammonium
sulfonato-a-D-qlucopyranosyllM1?4)-(methyl 3-O-benzyl-2-O-ammonium sulfonato-a-L-
idopvranosvluronate)-(1?4M2-azido-3-O-benzyl-2-deoxy-6-O-ammonium sulfonato--D-
D-glucopyranosvl)-(1?4)-(methyl______3-O-benzyl-2-O-ammonium______sulfonato-a-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosyl)-(1?4)-
(methyl 3-O-benzyl-2-O-ammonium sulfonato--D-i_-idopvranosvluronateM1?4)-3-O-
benzyl-2-deoxy-2-f(benzyloxy)carbonvnamino-B--D-qlucopyranoside (No. 164)
Compound 160 (196 mg, 0.048 mmol) is treated according to a protocol similar
to that described for the synthesis of 163. The reaction time may optionally be
shortened according to the TLC monitoring. Moreover, reverse-phase purification may
prove to be necessary in order to obtain compound 164 (104 mg, 59%).
Rf = 0.32 (57/29/7.2/16 EtOAc/pyridine/AcOH/H20).
Preparation of 3-cyclohexvlpropyl (methyl 3,4-di-O-benzyl-2-O-ammonium
sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-ammonium
sulfonato-a-D-qlucopyranosyllM1 -»4)-(methyl 3-O-benzyl-2-O-ammonium sulfonato-a-L-
idopvranosvluronate)-(1^4M2-azido-3-O-benzyl-2-deoxy-6-O-ammonium sulfonato--D-
D-qlucopyranosyll)-(1?4Mmethyl______3-O-benzyl-2-O-ammonium______sulfonato-a-L-
idopvranosvluronateH1-+4M2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosvlH1?4)-
(methvl 3-O-benzyl-2-O-ammonium sulfonato-a-L-idopyranosvluronate)-(1?4)-3-O-
benzyl-2-deoxy-2-r(benzyloxy)carbonyllamino-S--D-qlucopyranoside (No. 165)
Compound 161 (230 mg, 0.055 mmol) is treated according to a protocol similar
to that described for the synthesis of 164, to give compound 165 (115 mg, 56%).
Rf = 0.57 (57/29/7.2/16 EtOAc/pyridine/AcOH/H20).
Preparation of 3,3-diphenylpropvl (methyl 3,4-di-O-benzyl-2-O-ammonium
sulfonato-a-L-idopvranosvluronate)-(1^-4)-(2-azido-3-O-benzyl-2-deoxy-6-O-ammonium
sulfonato-a-D-qlucopyranosvl)-(1?4)-(methvl 3-O-benzyl-2-O-ammonium sulfonato-O-L-
idopvranosvluronateH1^4M2-azido-3-O-benzyl-2-deoxy-6-O-ammonium sulfonato--D-
D-qlucopyranosyllM1?4)-(methyl______3-O-benzyl-2-O-ammonium______sulfonato-g-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-a-D-qlucopyranosvl)-(1?4)-
(methyl 3-O-benzyl-2-O-ammonium sulfonato-g-L-idopyranosyluronateHI —>4)-3-O-
benzyl-2-deoxy-2-r(benzyloxy)carbonvnamino-B--D-qlucopyranoside (No. 166)
Compound 162 (43 mg, 10.2 umol) is treated according to a protocol similar to
that described for the synthesis of 163, to give compound 166.
Rf = 0.32 (29/13/3.4/7 EtOAc/pyridine/AcOH/H20).
Preparation of 5-phenylpentvl (lithium 3,4-di-O-benzyl-2-O-lithium sulfonato-a-L-
idopyranosvluronate)-(1?4M2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-O--D-
qlucopyranosylHI ->4Hlithium 3-O-benzyl-2-O-lithium sulfonato-O-L-idopyranosyl-
uronateVd—»4)-(2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-a-D-qlucopyranosvl)-
(1?4)-(lithium 3-O-benzyl-2-O-lithium sulfonato--D-i_-idopvranosvluronate)-(1?4)-(2-
azido-3-O-benzyl-2-deoxy-g--D-qlucopyranosvn-(1?4)-(lithium 3-O-benzyl-2-O-lithium
sulfonato-a-L-idopvranosvluronate)-(1?4)-3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonvfl-
amino-B--D-qlucopyranoside (No. 167)
To a solution of compound 163 (289 mg, 91 umol) in methanol (3.59 ml_) are
added, at 0°C, aqueous 30% hydrogen peroxide solution (2.23 mL), followed by
dropwise addition of aqueous 5N LiOH solution (1.46 mL). After stirring for 3 hours at
0°C, for 16 hours at room temperature and then for 27 hours at 45°C, the reaction
medium is neutralized at 0°C by addition of 3N hydrochloric acid, and then purified by
means of an LH-20 column, using a 9/1 mixture of methanol/ty/V-dimethylformamide as
eluent. The fractions containing the product are then concentrated under high vacuum
to give the desired compound 167 (267 mg, 95%).
Rf = 0.25 (57/29/7.2/16 EtOAc/pyridine/AcOH/H20).
Preparation of 2-propylpentyl (lithium 3,4-di-O-benzyl-2-O-lithium sulfonato-a-L-
idopvranosvluronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxyT6-O-lithium sulfonato-g--D-
qlucopyranosyllH1?4)-(lithium 3-O-benzyl-2-O-lithium sulfonato-o-L-idopyranosvl-
uronate)-(1?4H2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-a-D-qlucopyranosvl)-
(1—»4)-(lithium 3-O-benzyl-2-O-lithium sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-
azido-3-O-benzyl-2-deoxy-a-D-glucopyranosvlH1?4)-(lithium 3-O-benzyl-2-O-lithium
sulfonato-a-L-idopvranosvluronate)-(1?4)-3-O-benzyl-2-deoxy-2-[(benzyloxy)carbonyl1-
amino-3--D-qlucopyranoside (No. 168)
Compound 164 (104 mg, 0.029 mmol) is treated according to a protocol similar
to that described for the synthesis of 167, to give compound 168.
Rf = 0.28 (57/29/7.2/16 EtOAc/pyridine/AcOH/H20).
Preparation of 3-cvclohexylpropyl (lithium 3,4-di-O-benzyl-2-O-lithium sulfonato-
a-L-idopvranosvluronateH1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-a-D-
glucopvranosvD-(1?4Hlithium 3-O-benzyl-2-O-lithium sulfonato-a-L-idopyranosyl-
uronate)-(1—»4)-(2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-a-D-qlucopyranosvl)-
(1?4)-(lithium 3-O-benzyl-2-O-lithium sulfonato-a-L-idopyranosvluronate)-( 1 —^V-te-
azido-S-O-benzyl^-deoxy-O--D-qlucopyranosyllV-d—»4)-(lithium 3-O-benzyl-2-O-lithium
sulfonato-g-L-idopvranosvluronate)-(1?4)-3-O-benzyl-2-deoxy-2-r(benzyloxy)carbonyll-
amino-B--D-glucopyranoside (No. 169)
Compound 165 (110 mg, 0.030 mmol) is treated according to a protocol similar
to that described for the synthesis of 167, to give compound 169 (76 mg, 83%).
Rf = 0.20 (57/29/7.2/16 EtOAc/pyridine/AcOH/H20).
Preparation of 3,3-diphenvlpropyl (lithium 3,4-di-O-benzyl-2-O-lithium sulfonato-
Q-L-idopvranosvluronate)-(1—»4)-(2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-O--D-
qlucopyranosylH1?4Mlithium 3-O-benzyl-2-O-lithium sulfonato-O-L-idopyranosyl-
uronate)-(1?4)-(2-azido-3-O-benzyl-2-deoxy-6-O-lithium sulfonato-O--D-qlucopyranosyl)-
(1?4)-(lithium 3-O-benzyl-2-O-lithium sulfonato-g-L-idopyranosvluronate)-(1?4)-(2-
azido-3-O-benzyl-2-deoxy-g--D-glucopvranosyl)-(1^-4)-(lithium 3-O-benzyl-2-O-lithium
sulfonato-g-L-idopvranosvluronate)-(1?4)-3-O-benzyl-2-deoxy-2-f(benzyloxy)carbonvn-
amino-B--D-qlucopyranoside (No. 170)
Compound 166 is treated according to a protocol similar to that described for the
synthesis of 167. If necessary, the residue obtained may be used under the same
conditions to give compound 170 (27 mg, 87% (2 steps)).
Rf = 0.39 (57/29/7.2/16 EtOAc/pyridine/AcOH/H20).
Preparation of 5-phenvlpentvl (sodium 2-O-sodium sulfonato-O-L-idopyranosvl-
uronate)-(1?4)-(2-amino-2-deoxy-6-O-sodium sulfonato-a-D-qlucopyranosyll)-( 1?4 )-
(sodium 2-O-sodium sulfonato-O-L-idopvranosvluronateH1^4M2-amino-2-deoxy-6-O-
sodium sulfonato-a-D-qlucopyranosyll)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopvranosvluronateH1^4H2-amino-2-deoxy-O--D-qlucopvranosyl)-(1?4)-(sodium 2-
O-sodium______sulfonato-a-L-idopvranosvluronateM1?4)-2-amino-2-deoxy-3--D-qluco-
pyranoside (No. 171)
To a solution in a 1/1 mixture of terf-butanol/water (16.1 mL) of compound 167
obtained previously are successively added ammonium formate (658 mg, 10.4 mmol)
and then 10% Pd/C (1.61 g). After stirring vigorously for 4 hours at room temperature,
the reaction medium is filtered and partially concentrated under vacuum, and the
solution is then applied to a Sephadex® G-25 column eluted with 0.2 M sodium chloride.
The fractions containing the product are concentrated and desalified using the same
column eluted with water. The fractions containing the product are then concentrated
under high vacuum to give the desired compound 171 (109 mg, 62%).
"ESI" method, negative mode: multicharged ion detected m/z 663.08 [M-3H]3"
(acid form).
Preparation of 2-propylpentvl (sodium 2-O-sodium sulfonato-a-L-idopyranosvl-
uronate)-(1?4)-(2-amino-2-deoxy-6-O-sodium sulfonato-O--D-qlucopvranosyl)-(1 -+4)-
(sodium 2-O-sodium sulfonato-a-L-idopyranosvluronateH1?4)-(2-amino-2-deoxy-6-O-
sodium sulfonato-a-D-qlucopyranosyll)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopyranosvluronateHI —»4)-(2-amino-2-deoxy-a-D-qlucopyranosvl)-(1?4)-(sodium 2-
O-sodium______sulfonato-a-L-idopvranosvluronateH1—»4)-2-amino-2-deoxy-B--D-qluco-
pyranoside (No. 172)
Compound 168 is treated according to a protocol similar to that described for the
synthesis of 171, to give compound 172 (47 mg, 74% (2 steps)).
Chemical shifts of the anomeric protons (500 MHz, D20) 5 5.29* ldollAvl", 5.50**
Glcv", 5.29* ldoUAvl, 5.48** Glcv, 5.29* ldoUAlv, 5.48** Glc'", 5.22* IdoUA", 4.78** Glc1
* and **: The signals may be interchanged.
Preparation of 3-cvclohexvlpentyl (sodium 2-O-sodium sulfonato-a-L-
idopyranosvluronateM1?4V(2-amino-2-deoxy-6-O-sodium_______sulfonato-a-D-qluco-
pyranosvD-d—»4Hsodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-(1?4)-(2-
amino-2-deoxy-6-O-sodium sulfonato-o--D-qlucopyranosyll)-(1?4)-(sodium 2-O-sodium
sulfonato-O-L-idopvranosvluronateM1?4)-(2-amino-2-deoxy-a-D-qlucopyranosylO-
(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-(1?4)-2-amino-2-
deoxy-8--D-qlucopyranoside (No. 173)
Compound 169 (88 mg, 29 umol) is treated according to a protocol similar to that
described for the synthesis of 171, to give compound 173 (63 mg, 99%).
Chemical shifts of the anomeric protons (500 MHz, D20) 5 5.29* ldollAvl", 5.50**
Glcv", 5.29* ldoUAvl, 5.48** Glcv, 5.29* ldoUAlv, 5.48** Glc"', 5.22* IdoUA", 4.78** Glc1
* and **: The signals may be interchanged.
Preparation of 3,3-diphenylpentyl (sodium 2-O-sodium sulfonato-a-L-
idopvranosvluronateM1?4H2-amino-2-deoxy-6-O-sodium_______sulfonato-a-D-gluco-
pyranosvlV1?4)-(sodium 2-O-sodium sulfonato-a-L-idopvranosvluronateH1?4H2-
amino-2-deoxy-6-O-sodium sulfonato-g--D-qlucopvranosyl)-(1?4Hsodium 2-O-sodium
sulfonato-a-L-idopyranosvluronate)-(1—»4)-(2-amino-2-deoxy-a-D-qlucopyranosvl)-
(1—»4Hsodium 2-O-sodium sulfonato-a-L-idopvranosvluronateM1?4)-2-amino-2-
deoxy-B--D-glucopyranoside (No. 174)
Compound 170 (17.4 mg, 5.6 umol) is treated according to a protocol similar to
that described for the synthesis of 171, to give compound 174 (5.5 mg, 44%).
Chemical shifts of the anomeric protons (600 MHz, D20) 6 5.15 ldoUAvl", 5.42
Glcv", 5.23 ldoUAvl, 5.40 Glcv, 5.23 ldoUAlv, 5.38 Glc'", 5.19 IdoUA", 4.39 Glc'
Examples of compounds according to the invention:
EXAMPLE 1: Pentvl (sodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-
(1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-qlucopyranosyllH1?4)-(sodium
2-O-sodium_____sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-
sodium sulfonato-a-D-qlucopyranosyll)-(1?4)-(sodium 2-O-sodium sulfonato-o-L-
idopvranosvluronateH1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-qluco-
pvranosvl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosvluronateH1?4>2-
acetamido-2-deoxy-6-O-sodium sulfonato-B--D-glucopyranoside (Compound 1)

To a saturated aqueous sodium hydrogen carbonate solution (2.1 ml_) of
compound 43 (39 mg, 16.6 umol) is added sodium hydrogen carbonate (223 mg,
2.66 mmol), followed by dropwise addition, at 0°C, of acetic anhydride (126 uL,
1.33 mmol). After stirring vigorously for 3 hours at 0°C and then for 15 hours at room
temperature, the reaction medium is applied to a Sephadex® G-25 column eluted with
0.2 M sodium chloride. The fractions containing the product are concentrated and
desalified using the same column eluted with water. The fractions containing the
product are then concentrated under high vacuum to give the desired compound 1
(30 mg, 73%).
Chemical shifts of the anomeric protons (500 MHz, D20) 5 5.14 ldoUAvl", 5.13
Glcv", 5.16 ldoUAvl, 5.13 Glcv, 5.15 ldoUAlv, 5.14 Glc'", 5.17 IdoUA", 4.51 Glc1
Mass: "ESI" method, negative mode: theoretical mass = 2509.72; experimental
mass: 2508.82 ± 0.34 a.m.u.
EXAMPLE 2: Pentvl (sodium 2-O-sodium sulfonato-g-L-idopyranosvluronate)-
(1?4H2-deoxy-2-(1-oxobutv0amino-6-O-sodium sulfonato-a-D-qlucopyranosvl)-(1?4)-
(sodium 2-O-sodium sulfonato-a-L-idopvranosvluronateH1?4M2-deoxy-2-(1-oxobutyl)-
amino-6-O-sodium sulfonato-a-D-glucopyranosvlH1^>4)-(sodium 2-O-sodium sulfonato-
a-L-idopyranosvluronate)-(1 H>4H2-deoxy-2-(1 -oxobutyl)amino-6-O-sodium sulfonato--D-
D-glucopyranosvD-(1?4Hsodium 2-O-sodium sulfonato-a-L-idopvranosvluronate)-
(1?4)-2-deoxy-2-(1-oxobutyl)amino-6-O-sodium________sulfonato-B--D-glucopvranoside
(Compound 2)

To a solution of compound 43 (26 mg, 11.1 umol) in a 7/3 mixture of N,N-
dimethylformamide/water (2.9 ml_) are added dropwise, at 0°C, diisopropylethylamine
(33 uL, 17 molar equivalents) dissolved in A/,A/-dimethylformamide (100 pL) and N-
hydroxysuccinimide butyrate (27.4 mg, 13 molar equivalents) dissolved in N,N-
dimethylformamide (100 uL). Stirring is continued at room temperature for 4 hours, and
the same amount of reagent is then added twice more under the same conditions. After
a total of 18 hours of stirring at room temperature, the reaction medium is applied to a
Sephadex® G-25 column eluted with 0.2 M sodium chloride. The fractions containing
the product are concentrated and desalified using the same column eluted with water.
The fractions containing the product are then concentrated under high vacuum to give
the desired compound 2 (26 mg, 89%).
Chemical shifts of the anomeric protons (600 MHz, D20) 8 5.14 ldoUAvl", 5.12*
Glcv", 5.14 ldoUAvl, 5.11* Glcv, 5.14 IdoUA^, 5.10* Glc'", 5.15 IdoUA", 4.50 Glc1
*: The signals may be interchanged.
"ESI" method, negative mode: multicharged ion detected m/z 588.1095 [M-4H]4*
(acid form).
EXAMPLE 3: Pentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-
(1?4)-(2-deoxy-2-(1-oxohexvDamino-6-O-sodium sulfonato-a-D-qlucopyranosvlV(1?4)-
(sodium 2-O-sodium sulfonato-a-L-idopyranosvluronateH1?4M2-deoxy-2-(1-
oxohexvOamino-6-O-sodium sulfonato-a-D-qlucopyranosvO-(1?4)-(sodium 2-O-sodium
sulfonato-a-L-idopvranosvluronateH1^4M2-deoxy-2-(1-oxohexv0amino-6-O-sodium
sulfonato-a-D-qlucopyranosylO-(1?4)-(sodium 2-O-sodium sulfonato-o-L-idopyranosyl-
uronateH1?4y-2-deoxy-2-(1-oxohexvDamino-6-O-sodium________sulfonato-B--D-qluco-
pyranoside (Compound 3)

To a solution of compound 43 (10 mg, 4.27 umol) in water (0.3 ml_) are added
dropwise, at OX, diisopropylethylamine (20 uL, 26.8 molar equivalents) dissolved in
A/,/v-dimethylformamide (200 uL) and N-hydroxysuccinimide hexanoate (18.2 mg,
20 molar equivalents) dissolved in A/,A/-dimethylformamide (500 uL). Stirring is
continued at 0°C for 15 minutes and then at room temperature for 16 hours. The
reaction medium is then applied to a Sephadex® G-25 column eluted with 0.2 M sodium
chloride. The fractions containing the product are concentrated and desalified using the
same column eluted with water. The fractions containing the product are then
concentrated under high vacuum to give the desired compound 3 (4 mg, 34%).
Chemical shifts of the anomeric protons (600 MHz, D20) 8 5.11 ldoUAvl", 5.08
Glcv", 5.14 ldoUAvl, 5.09 Glcv, 5.13 ldoUAlv, 5.10 Glc"', 5.14 IdollA", 4.47 Glc1
"ESI" method, negative mode: multicharged ion detected m/z 627.6120 [M-4H]4"
(acid form).
EXAMPLE 4: Pentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-
(1?4H2-r(cvclopentvlcarbonv0amino1-2-deoxy-6-O-sodium_______sulfonato-a-D-qluco-
pyranosvlH1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-(1?4H2-
r(cvclopentvlcarbonyl)aminol-2-deoxy-6-O-sodium sulfonato-a-D-qlucopyranosvD-(1?4V
(sodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-(1 -^4)-(2-r(cvclopentyl-
carbonyl)aminol-2-deoxy-6-O-sodium sulfonato-O--D-qlucopyranosyll)-(1?4)-(sodium 2-
O-sodium sulfonato-a-L-idopvranosvluronateH1^4V2-r(cvclopentvlcarbonvnamino1-2-
deoxy-6-O-sodium sulfonato-B--D-glucopyranoside (Compound 4)
To a solution of compound 43 (9 mg, 3.84 umol) in water (0.3 mL) are added
dropwise, at 0°C, A/,A/-dimethylformamide (0.5 mL), diisopropylethylamine (7 uL,
2.6 molar equivalents per free amine) dissolved in A/,/v"-dimethylformamide (50 uL) and
N-hydroxysuccinimide cyclopentanecarboxylate (6.6 mg, 2 molar equivalents per free
amine) dissolved in A/,A/-dimethylformamide (50 uL). Stirring is continued at room
temperature for 2 hours, and the same amount of reagent is then added four more
times under the same conditions. After a total of 18 hours of stirring at room
temperature, the reaction medium is applied to a Sephadex® G-25 column eluted with
0.2 M sodium chloride. The fractions containing the product are concentrated and
desalified using the same column eluted with water. The fractions containing the
product are then concentrated under high vacuum to give the desired compound 4
(5.6 mg, 55%). In the event of an incomplete reaction, the product obtained may be
reused under the same conditions. If necessary, it may be purified by ion-exchange
chromatography using a semi-preparative Dionex CarboPac® PA100 column (9 x
250 mm) with a gradient starting from an eluent A to an eluent B, eluent A consisting of
a water/acetonitrile mixture (4/1) + 0.01% of dimethyl sulfoxide, and eluent B consisting
of a mixture of a 2N sodium chloride solution and acetonitrile (4/1). The fractions
containing the product are then concentrated and desalified using a Sephadex® G-25
column eluted with water. The fractions containing the product are then concentrated
under high vacuum to give the desired compound 4.
Chemical shifts of the anomeric protons (600 MHz, D20) 8 5.06* ldoUAvl", 5.05**
GlcVM, 5.08* ldoUAvl, 5.03** Glcv, 5.06* ldoUAlv, 5.05** Glc"', 5.08* IdoUA", 4.44** Glc1
* and **: The signals may be interchanged.
[a]D 15.4° (c 0.25; H20).
EXAMPLE 5: Undecvl (sodium 2-O-sodium sulfonato-a-L-idopvranosvluronate)-
(1?4M2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-qlucopyranosyllMW4)-(sodium
2-O-sodium_____sulfonato-a-L-idopyranosvluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-
sodium sulfonato-g--D-qlucopyranosyllM1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopyranosvluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-gluco-
pvranosvD-(1?4V(sodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-(1?4)-2-
acetamido-2-deoxy-6-O-sodium sulfonato-B--D-glucopyranoside (Compound 5)
To a saturated aqueous sodium hydrogen carbonate solution (2.3 mL) of
compound 52 (32 mg, 13.2 umol) is added sodium hydrogen carbonate (174 mg,
2.08 mmol), followed by dropwise addition, at 0°C, of acetic anhydride (100 uL,
1.05 mmol). After stirring vigorously for 2.5 hours at 0°C and then for 15 hours at room
temperature, the reaction medium is applied to a Sephadex® G-25 column eluted with
0.2 M sodium chloride. The fractions containing the product are concentrated and
desalified using the same column eluted with water. The fractions containing the
product are then concentrated under high vacuum to give the desired compound 5
(31 mg, 91%).
Chemical shifts of the anomeric protons (500 MHz, D20) 5 5.14 idoUAvl", 5.13*
Glcv", 5.14** ldoUAvl, 5.13* Glcv, 5.15** ldoUAlv, 5.13* Glc'", 5.16 IdoUA", 4.50 Glc1
* and **: The signals may be interchanged.
[a]D18.6°(c1;H20).
EXAMPLE 6: 5-phenylpentvl (sodium 2-O-sodium sulfonato-a-L-idopyranosvl-
uronateH1?4H2-acetamido-2-deoxy-6-O-sodium______sulfonato-a-D-glucopyranosvl)-
(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopvranosyluronateM1?4M2-acetamido-2-
deoxy-6-O-sodium sulfonato-a-D-glucopvranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-
a-L-idopvranosvluronateH1?4M2-acetamido-2-deoxy-6-O-sodium_____sulfonato-a-D-
qlucopyranosvl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopvranosvluronate)-(1^4V
2-acetamido-2-deoxy-6-O-sodiumsulfonato-3--D-qlucopyranoside (Compound 6)

To a saturated aqueous sodium hydrogen carbonate solution (2.7 mL) of
compound 53 (36 mg, 14.8 umol) is added sodium hydrogen carbonate (199 mg,
2.38 mmol), followed by dropwise addition, at 0°C, of acetic anhydride (112 uL,
1.19 mmol). After stirring vigorously for 2.5 hours at 0°C and then for 15 hours at room
temperature, the reaction medium is applied to a Sephadex® G-25 column eluted with
0.2 M sodium chloride. The fractions containing the product are concentrated and
desalified using the same column eluted with water. The fractions containing the
product are then concentrated under high vacuum to give the desired compound 6
(25.6 mg, 68%). If necessary, it may be purified by ion-exchange chromatography using
a semi-preparative Dionex CarboPac® PA100 column (9 x 250 mm) with a gradient
starting from an eluent A to an eluent B, eluent A consisting of a water/acetonitrile
mixture (4/1) + 0.01% of dimethyl sulfoxide, and eluent B consisting of a mixture of a 2N
sodium chloride solution and acetonitrile (4/1). The fractions containing the product are
then concentrated and desalified using a Sephadex® G-25 column eluted with water.
The fractions containing the product are then concentrated under high vacuum to give
the desired compound 6.
Chemical shifts of the anomeric protons (500 MHz, D20) 5 5.14 ldoUAvl", 5.12
Glcv", 5.16 ldoUAvl, 5.12 Glcv, 5.16 IdoUA^, 5.12 Glc'", 5.17 IdoUA", 4.48 Glc'
"ESI" method, negative mode: multicharged ion detected m/z 579.0867 [M-4H]4"
(acid form).
EXAMPLE 7: Methyl (sodium 2-O-sodium sulfonato-O-L-idopyranosyluronate)-
(1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-qlucopvranosyl)-f(1?4)-
(sodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-(1?4M2-acetamido-2-deoxy-
6-O-sodium sulfonato-O--D-qlucopyranosylM7-(1?4Hsodium 2-O-sodium sulfonato-a-L-
idopvranosvluronate)-(1?4)-2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-qluco-
pyranoside (Compound 7)

To a solution of compound 77 (10.8 mg, 4.73 umol) in saturated sodium
hydrogen carbonate solution (472 uL) are successively added, at 0°C and under an
inert atmosphere, solid sodium hydrogen carbonate (64 mg) and acetic anhydride
(36 uL). After stirring for 18 hours at room temperature, the reaction mixture is filtered
(Millipore® LSWP 5 urn filter) and then applied to a column of fine Sephadex® G-25 gel
eluted with aqueous 0.2 M NaCI solution. The fractions containing the expected product
are pooled and applied to a column of fine Sephadex® G-25 (95 x 2 cm) eluted with
water, to give 6.7 mg of compound 7.
1H NMR (D20) 5 of the anomeric protons: 5.20; 5.18 (2 H); 5.17 (2H); 5.16; 5.15;
4.78 ppm.
Mass: "ESI" method, negative mode: theoretical mass = 2453.62; experimental
mass: 2453.22 + 0.17 a.m.u..
EXAMPLE 8: Methyl (sodium 2-O-sodium sulfonato-g-L-idopyranosyluronate)-
(1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-g--D-glucopvranosvl)-(1?4)-(sodium
2-O-sodium_____sulfonato-g-L-idopvranosvluronate)-(1?4H2-acetamido-2-deoxy-6-O-
sodium sulfonato-a-D-qlucopvranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopvranosvluronate)-(1?4)-(2-acetamido-2-deoxy-a-D-glucopyranosvn-(1?4)-(sodium
2-O-sodium______sulfonato-a-L-idopyranosvluronate)-(1?4)-2-acetamido-2-deoxy-6-O-
sodiumsulfonato-g--D-glucopyranoside (Compound 8)

Compound 85 (23.0 mg, 11.1 umol) is treated according to the same procedure
as that described for the preparation of compound 7, to give compound 8 (25.2 mg).
1H NMR (D20) 6 of the anomeric protons: 5.45; 5.42 (2 H); 5.22; 5.21; 5.20;
5.18; 5.03 ppm.
Mass: "ESI" method, negative mode: theoretical mass = 2249.53; experimental
mass: 2249.19 ± 0.18 a.m.u.
EXAMPLE 9: Methyl (sodium 2-O-sodium sulfonato-a-L-idopvranosvluronate)-
(1?4M2-acetamido-2-deoxy-6-O-sodium sulfonato-O--D-qlucopyranosyllV(1?4Hsodium
2-O-sodium_____sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-
sodium sulfonato-a-D-glucopyranosvl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopvranosvluronateV(1?4)-(2-acetamido-2-deoxy-O--D-glucopyranosvl)-(1?4)-(sodium
2-O-sodium sulfonato-g-L-idopvranosvluronateH1^4)-2-acetamido-2-deoxy-g--D-qluco-
pyranoside (Compound 9)

Compound 90 (52.1 mg, 23.9 umol) is treated according to the same procedure
as that described for the preparation of Example 7, to give compound 9 (50.7 mg).
1H NMR (D20) 8 of the anomeric protons: 5.18; 5.16 (2H); 5.14; 5.15; 5.13; 5.09;
4.75 ppm.
Mass: "ESI" method, negative mode: theoretical mass = 2351.57; experimental
mass: 2350.88 ± 0.10 a.m.u.
EXAMPLE 10: Methyl (sodium 2-O-sodium sulfonato-a-L-idopvranosvluronate)-
(1?4)-(2-acetamido-2-deoxy-a-D-glucopvranosvlH1?4Hsodium_________2-O-sodium
sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-
a--D-glucopyranosvl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-
(1?4H2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-qlucopyranosyllH1 -^4)-(sodium
2-O-sodium sulfonato-a-L-idopvranosvluronate)-(1?4)-2-acetamido-2-deoxy-a-D-qluco-
pyranoside (Compound 10)

To a saturated aqueous sodium hydrogen carbonate solution (3.7 ml_) of
compound 100 (76 mg, 37 umol) is added sodium hydrogen carbonate (497 mg,
5.92 mmol), followed by dropwise addition, at 0°C, of acetic anhydride (283 uL,
3.0 mmol). After stirring vigorously for 3 hours at 0°C and then for 15 hours at room
temperature, the reaction medium is applied to a Sephadex® G-25 column eluted with
0.2 M sodium chloride. The fractions containing the product are concentrated and
desalified using the same column eluted with water. The fractions containing the
product are then concentrated under high vacuum to give the desired compound 10
(61 mg, 73%).
Chemical shifts of the anomeric protons (600 MHz, D20) 8 5.13 ldoUAvl", 5.13
Glcv", 5.15 ldoUAvl, 5.13 Glcv, 5.16 ldoUAlv, 5.10 Glc'", 5.16 IdoUA", 4.75 Glc1
Mass: "ESI" method, negative mode: theoretical mass = 2249.53; experimental
mass: 2249.02 ± 0.35 a.m.u.
EXAMPLE 11: Methyl (sodium 2-O-sodium sulfonato-a-L-idopvranosyluronate)-
(1?4)-(2-acetamido-2-deoxy-O--D-qlucopyranosyll)-(1?4)-( sodium_________2-O-sodium
sulfonato-a-L-idopvranosvluronateH1?4H2-acetamido-2-deoxy-6-O-sodium sulfonato-
q--D-qlucopyranosyll)-(1?4Hsodium 2-O-sodium sulfonato-a-L-idopvranosvluronate)-
(1?4M2-acetamido-2-deoxy-6-O-sodiunn sulfonato-a-D-qlucopyranosyllH1?4)-(sodium
2-O-sodium______sulfonato-a-L-idopvranosvluronate)-(1?4)-2-acetamido-2-deoxy-6-O-
sodium sulfonato-g--D-qlucopyranoside (Compound 11)

To a saturated aqueous sodium hydrogen carbonate solution (2.6 mL) of
compound 105 (56 mg, 26 umol) is added sodium hydrogen carbonate (349 mg,
4.16 mmol), followed by dropwise addition, at 0°C, of acetic anhydride (194 uL,
2.05 mmol). After stirring vigorously for 3 hours at 0°C and then for 15 hours at room
temperature, the reaction medium is applied to a Sephadex® G-25 column eluted with
0.2 M sodium chloride. The fractions containing the product are concentrated and
desalified using the same column eluted with water. The fractions containing the
product are then concentrated under high vacuum to give the desired compound 11
(45 mg, 74%).
Chemical shifts of the anomeric protons (600 MHz, D20) 8 5.13 ldoUAvl", 5.12
Glcv", 5.16 ldollAvl, 5.12 Glcv, 5.15 ldoUAlv, 5.13 Glc'", 5.17 IdoUA", 4.72 Glc'
Mass: "ESI" method, negative mode: theoretical mass = 2351.57; experimental
mass: 2351.13 ±0.06 a.m.u.
EXAMPLE 12: Methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-
(1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-qlucopyranosv0-(1-»4V-(sodium
2-O-sodium_____sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-acetamido-2-deoxy-a-D-
qlucopyranosvlMI ->4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosvluronate')-(1 -»4V
(2-acetamido-2-deoxy-6-O-sodium sulfonato-O--D-qlucopyranosvl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopvranosyluronate)-(1—»4)-2-acetamido-2-deoxy-a-D-qluco-
pyranoside (Compound 12)
To a saturated aqueous sodium hydrogen carbonate solution (1.4 mL) of
compound 113 (29.5 mg, 14 umol) is added sodium hydrogen carbonate (188 mg,
2.24 mmol), followed by dropwise addition, at 0°C, of acetic anhydride (105 uL,
1.12 mmol). After stirring vigorously for 3 hours at 0°C and then for 15 hours at room
temperature, the reaction medium is applied to a Sephadex® G-25 column eluted with
0.2 M sodium chloride. The fractions containing the product are concentrated and
desalified using the same column eluted with water. The fractions containing the
product are then concentrated under high vacuum to give the desired compound 12
(29 mg, 92%).
Chemical shifts of the anomeric protons (600 MHz, D20) 8 5.12 ldoUAvl", 5.06
Glcv", 5.14 ldoUAvl, 5.11 Glcv, 5.13 ldoUAlv, 5.09 Glc'", 5.14 IdoUA", 4.73 Glc'
[a]D 115.3° (c 0.3; H20).
EXAMPLE 13: Methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-
(1?4H2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-qlucopyranosylM1?4Msodium
2-O-sodium_____sulfonato-a-L-idopvranosvluronateM1?4M2-acetamido-2-deoxy-a-D-
qlucopyranosvlM1?4Hsodium 2-O-sodium sulfonato-a-L-idopvranosvluronateM1?4)-
(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-qlucopyranosyll)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopvranosvluronate)-(1?4)-2-acetamido-2-deoxy-6-O-sodium
sulfonato-g--D-qlucopyranoside (Compound 13)

To a saturated aqueous sodium hydrogen carbonate solution (1.26 mL) of
compound 118 (27 mg, 12.6 umol) is added sodium hydrogen carbonate (169 mg,
2.02 mmol), followed by dropwise addition, at 0°C, of acetic anhydride (95 uL,
1.01 mmol). After stirring vigorously for 3 hours at 0°C and then for 16 hours at room
temperature, the reaction medium is applied to a Sephadex® G-25 column eluted with
0.2 M sodium chloride. The fractions containing the product are concentrated and
desalified using the same column eluted with water. The fractions containing the
product are then concentrated under high vacuum to give the desired compound 13
(26.5 mg, 89%).
Chemical shifts of the anomeric protons (600 MHz, D20) 5 5.12 ldoUAvl", 5.05
Glcv", 5.13 ldoUAvl, 5.10 Glcv, 5.13 ldoUAlv, 5.10 Glc'", 5.14 IdoUA", 4.72 Glc1
Mass: "ESI" method, negative mode: theoretical mass = 2351.57; experimental
mass: 2350.96 ± 0.20 a.m.u.
EXAMPLE 14: 5-phenylpentvl (sodium 2-O-sodium sulfonato-a-L-idopvranosvl-
uronate)-(1?4M2-deoxy-2-(1-oxobutv0amino-6-O-sodium________sulfonato-a-D-qluco-
pyranosylM1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosvluronateM1-»4M2-
deoxy-2-(1-oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium
2-O-sodium sulfonato-a-L-idopvranosvluronateH1 -^4M2-deoxy-2-(1 -oxobutyl)amino--D-
D-qlucopvranosyl)-(1?4Msodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-
(1?4)-2-deoxy-2-(1-oxobutyl)amino-B--D-qlucopyranoside (Compound 14)

To a solution of compound 171 (126 mg, 56.9 umol) in a 7/3 mixture of N,N-
dimethylformamide/water (12.6 ml_) are added dropwise, at 0°C, diisopropylethylamine
(148 uL, 14.9 molar equivalents) dissolved in /V,/v"-dimethylformamide (100 uL), and N-
hydroxysuccinimide butyrate (116 mg, 11 molar equivalents) dissolved in N,N-
dimethylformamide (100 uL). Stirring is continued at room temperature for 3 hours, and
the same amount of reagent is then added three times under the same conditions. After
a total of 23 hours of stirring at room temperature, the reaction medium is applied to a
Sephadex® G-25 column eluted with 0.2 M sodium chloride. The fractions containing
the product are concentrated and desalified using the same column eluted with water.
The fractions containing the product are then concentrated under high vacuum to give
the desired compound 14 (133 mg, 93%).
Chemical shifts of the anomeric protons (600 MHz, D20) 8 5.07 ldoUAvl", 4.99
Glcv", 5.09 ldoUAvl, 5.04 Glcv, 5.10 ldoUAlv, 5.01 Glc'", 5.06 IdoUA", 4.38 Glc'
[a]D50.5°(c1;H2O).
EXAMPLE 15: 2-propvlpentvl (sodium 2-O-sodium sulfonato-a-L-idopyranosvl-
uronate)-(1?4M2-deoxy-2-(1-oxobutyl)amino-6-O-sodium________sulfonato-a-D-qluco-
pyranosyl)(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
-deoxy-2-(1-oxobutyl)amino-6-O-sodium sulfonato-O--D-qlucopvranosyl)-(1?4)-(sodium
2-O-sodium sulfonato-a-L-idopvranosvluronateV(1?4)-(2-deoxy-2-(1-oxobutyl)amino--D-
D-qlucopvranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopvranosvluronate)-
(1?4)-2-deoxy-2-(1-oxobutyl)amino-ß-D-glucopvranoside (Compound 15)

Compound 172 (47 mg, 22 umol) is treated according to a protocol similar to that
described for the synthesis of 14, to give compound 15 (46 mg, 85%).
Chemical shifts of the anomeric protons (600 MHz, D20) 5 5.33 ldoUAvl", 5.30
Glcv", 5.35 ldoUAvl, 5.24 Glcv, 5.36 ldoUAlv, 5.27 Glc'", 5.32 IdoUA", 4.63 Glc1
[a]D60°(c1;H2O).
EXAMPLE 16: 3-cvclohexylpropyl (sodium 2-O-sodium sulfonato-a-L-
idopvranosvluronate)-(1?4)-(2-deoxy-2-acetamido-6-O-sodium sulfonato-a-D-qluco-
pyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-(1?4))-(2-
deoxy-2-acetamido-6-O-sodium sulfonato-a-D-qlucopyranosvl)-(1?4)-(sodiurn 2-O-
sodium sulfonato-a-L-idopvranosyluronate)-(1?4)-(2-deoxy-2-acetamido-a-D-qluco-
pyranosvlM1?4Msodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-(1?4)-2-
deoxy-2-acetamido-ß-D-qlucopvranoside (Compound 16)

Compound 172 (10 mg, 4.56 umol) is treated according to a protocol similar to
that described for the synthesis of 13, to give compound 16 (8.8 mg, 82%).
Chemical shifts of the anomeric protons (600 MHz, D2O) 8 5.15 ldoUAvl", 5.08
Glcv", 5.17 ldoUAvl, 5.13 Glcv, 5.17 ldoUAlv, 5.11 Glc'", 5.16 IdoUA", 4.50 Glc1
[a]D +40.9° (c 0.88; HzO).
EXAMPLE 17: 3-cvclohexvlpropyl (sodium 2-O-sodium sulfonato-a-L-
idopvranosvluronate)-(1?4)-(2-deoxy-2-(1-oxobutyl)amino-6-O-sodium sulfonato-a-D-
qlucopvranosyl)1?4)-(sodium 2-O-sodium sulfonato-a-L-idopvranosvluronate)-(1?4)-
(2-deoxy-2-(1-oxobutyl)amino-6-O-sodium_______sulfonato-a-D-qlucopvranosyl)-(1?4)-
(sodium 2-O-sodium sulfonato-a-L-idopvranosvluronate)-(1?4M2-deoxy-2-(1-oxobutvO-
amino-a-D-qlucopvranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronate)-(1?4)-2-deoxy-2-(1-oxobutyl)amino-ß--D-qlucopyranoside (Compound 17)

Compound 173 (10 mg, 4.56 umol) is treated according to a protocol similar to
that described for the synthesis of 14, to give compound 17 (7.7 mg, 69%).
Chemical shifts of the anomeric protons (600 MHz, D20) 5 5.23 ldoUAvl", 5.20
Glcv", 5.25 ldoUAvl, 5.15 Glcv, 5.26 ldoUAlv, 5.17 Glc'", 5.22 IdoUA", 4.57 Glc1
[a]D +53.8° (c 0.77; H20).
EXAMPLE 18: 3.3-diphenvlpropyl (sodium 2-O-sodium sulfonato-a-L-
idopvranosyluronate)-(1?4)-(2-deoxy-2-(1 -oxobutyl)amino-6-O-sodium sulfonato-g-p-
qlucopvranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosvluronate)-(1?4)-
(2-deoxy-2-(1-oxobutvBamino-6-O-sodium_______sulfonato-a-D-qlucopyranosyll)-(1?4)-
(sodium 2-O-sodium sulfonato-a-L-idopvranosvluronate)-(1?4)-(2-deoxy-2-(1 -oxobutyl)-
amino-a-D-qlucopyranosyll)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronate)-(1?4)-2-deoxy-2-(1-oxobutyl)amino-ß--D-qlucopvranoside (Compound 18)

To a solution of compound 174 (5.5 mg, 2.4 umol) in a 2/1 mixture of N,N-
dimethylformamide/water (4.8 ml_) are added dropwise, at 0°C, diisopropylethylamine
(63 uL, 15 molar equivalents) dissolved in N,N-dimethylformamide (43 uL), and N-
hydroxysuccinimide butyrate (49 mg, 11 molar equivalents) dissolved in N,N-
dimethylformamide (43 uL). Stirring is continued at room temperature for 3 hours, and
the same amount of reagent is then added three times under the same conditions. After
a total of 24 hours of stirring at room temperature, the reaction medium is applied to a
Sephadex® G-25 column eluted with 0.2 M sodium chloride. The fractions containing
the product are concentrated and desalified using the same column eluted with water.
The fractions containing the product are then concentrated under high vacuum to give
the desired compound 18 (6 mg, 98%).
Chemical shifts of the anomeric protons (600 MHz, D20) 5 5.22 ldoUAvm, 5.19
Glcv", 5.23 ldoUAvl, 5.13 Glcv, 5.23 ldoUAlv, 5.17 Glc'", 5.20 IdoUA", 4.48 Glc1
[alD +24° (c 0.48, H20).
EXAMPLE 19: 5-phenvlpentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosvl-
uronate)-(1?4)-(2-deoxy-2-(3-methyl-1-oxobutyl)amino-6-O-sodium_____sulfonato-a-D-
qlucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopvranosvluronate)-(1?4)-
(2-deoxy-2-(3-methvl-1-oxobutv0amino-6-O-sodium sulfonato-a-D-qlucopyranosyQ-
(1?4Hsodium 2-O-sodium sulfonato-a-L-idopvranosvluronateH1?4M2-deoxy-2-(3-
methyl-1-oxobutyl)amino-a-D-qlucopyranosylD-(1?4Hsodium 2-O-sodium sulfonato--D-
L-idopvranosvluronate)-(1?4)-2-deoxy-2-(3-methvl-1-oxobutyl)amino-ß--D-qluco-
pyranoside (Compound 19)

A solution of compound 171 (84 mg, 37.8 umol) in a 7/3 mixture of N,N-
dimethylformamide/water (8.4 mL) is treated with 1-[(3-methylbutanoyl)oxy] pyrrolidine-
2,5-dione according to a process similar to the synthesis of compound 14, to give the
desired compound 19 (41.8 mg, 43%). If necessary, it may be purified by ion-exchange
chromatography using a semi-preparative Dionex CarboPac® PA100 column (9 x
250 mm) with a gradient starting from an eluent A to an eluent B, eluent A consisting of
a water/acetonitrile mixture (4/1) + 0.01% of dimethyl sulfoxide, and eluent B consisting
of a mixture of a 2N sodium chloride solution and acetonitrile (4/1). The fractions
containing the product are then concentrated and desalified using a Sephadex® G-25
column eluted with water. The fractions containing the product are then concentrated
under high vacuum to give the desired pure compound 19.
Chemical shifts of the anomeric protons (600 MHz, D20) 8 5.23 ldoUAvl", 5.19
Glcv", 5.24 ldoUAvl, 5.13 Glcv, 5.25 ldoUAlv, 5.17 Glc'", 5.22 IdoUA", 4.55 Glc1
"ESI" method, negative mode: multicharged ion detected m/z 581.3517 [M-4H]4"
(acid form).
The compounds according to the invention underwent pharmacological trials to
determine their agonist effect on the FGF receptors and their activity on angiogenesis
and also on post-ischaemic revascularization.
Model of in vitro angiogenesis; specific activity towards FGF2
The in vitro angiogenesis model corresponds to a rearrangement of human
venous endothelial cells on a biological matrix. The matrix is made by dispensing, into
each well of a 96-well plate (Becton Dickinson 353872), 60 pi of Matrigel® diluted to 1/3
(Growth factor reduced Matrigel®: Becton Dickinson 356230) in collagen (rat tail
collagen, type I: Becton Dickinson 354249). The biological matrix hardens after 1 hour
at 37°C.
Human venous endothelial cells (HUVEC ref: C-12200 - Promocell) are seeded
onto the biological matrix at 7800 cells/well in 120 pi of EBM® medium (Endothelial
Basal Medium, Lonza C3121) + 2% FCS (foetal calf serum - Lonza) + hEGF
(Recombinant Human Epidermal Growth Factor - Lonza) 10 ug/ml. The cells are
stimulated with FGF2 (R&D Systems / 234 - FSE - 0 50) 10 ng/ml or with the products of
the invention for 18 hours at 37°C in the presence of 5% CO2. After 24 hours, the cells
are observed under a microscope (X4 objective lens) and analysis of the length of the
pseudo-tubules is performed with the aid of image software (Biocom VisioLab 2000
software).
In this test of in vitro angiogenesis, the compounds of the invention have a
specific activity of between 10-6 M and 10-12M. For example, compounds 1 and 2 are
active at 10-11M.
Model of cellulose implant in mice
This model is an adaptation of the model described by Andrade et al.
(Microvascular Research, 1997, 54, 253-61) for testing pharmacological products
capable of activating the onset of angiogenesis.
The animals (white consanguineous BALB/c J mice) are anaesthetized with a
xylazine (Rompun®, 10 mg/kg)/ketamine (Imalgene® 1000, 100mg/kg) mixture
intraperitoneally. The animal's back is shaved and disinfected with Hexomedine®. An air
pocket is created subcutaneously on the mouse's back by injecting 5 ml of sterile air. An
incision of about 2 cm at the top of the animal's back is made in order to introduce a
sterile cellulose implant (disk 1 cm in diameter, 2 mm thick, Cellspon® ref. 0501)
impregnated with 50 µl of sterile solution containing the test product. The incision is then
sutured and cleaned with Hexomedine®.
On the days following the insertion of the implant, the mice can receive the
product into the implant via an injection through the skin (50 µl/implant/day) under
gaseous anaesthesia (5% isoflurane (Aerrane®, Baxter)).
Seven days after inserting the sponge, the mice are sacrificed by means of a
lethal dose of pentobarbital sodium (CEVA Sante Animate), administered
intraperitoneally. The skin is then excised, about 1 cm around the sponge, while
avoiding the scar, so as to release the skin and the sponge. The sponge is then cut into
several pieces and placed in a Ribolyser® containing 1 ml of lysis buffer (Cell Death
Detection ELISA, Roche). The tubes are shaken four times consecutively, for
20 seconds, at force 4, using a cell mill (FastPrep® FP 120). The tubes are then
centrifuged for 10 minutes at 2000xg at 20°C and the supernatants are frozen at -20°C
until the time of the haemoglobin assay. On the day of the assay, the tubes are again
centrifuged after thawing and the haemoglobin concentration is measured with the
Drabkin reagent (Sigma, volume for volume) by reading on a spectrophotometer at
405 nm against a standard range of bovine haemoglobin (Sigma).
The haemoglobin concentration in each sample is expressed in mg/ml from the
polynomial regression produced from the range. The results are expressed as a mean
value (± sem) for each group. The differences between the groups are tested with an
ANOVA followed by a Dunnett test on the square root of the values.
In this in vivo test, the compounds of the invention revealed a specific activity of
between 5 and 45 ng/site. For example, compounds 1 and 2 are active at 45 ng/site,
and compound 7 is active at 15 ng/site.
For comparative purposes relative to a compound analogous to compound 7
according to the invention, but in which all the -NH-acyl groups are replaced with -NH-
SO3- groups, compound 7 according to the invention has in this test an activity of
greater than 21% relative to the said fully sulfated analogue, which shows the
importance of the N-acyl groups in the compounds according to the invention.
Pharmacoki netics
The pharmacokinetic profile of the compounds according to the invention is
evaluated after cutaneous administration, to OF1 or C57/BL6 mice, at a concentration of
30 mg/kg. The systemic exposure of the compounds is evaluated after a single
administration to the animal anaesthetized beforehand with pentobarbital. The blood is
collected from the vena cava and then transferred into plastic tubes containing lithium
heparin. After centrifugation (2000xg for 10 minutes at 8°C), the concentration of the
compounds in the plasma is quantified by LC/MS-MS.
In general, the test compounds have an improved pharmacokinetic profile
(especially higher AUC and plasmatic concentration, and longer half-lives) relative to
fully sulfated analogous compounds (replacement of the N-acyl groups with -NH-SO3-
groups). By way of example, the pharmacokinetic parameters of compounds 1, 6 and 7
according to the invention are described in the table that follows, in comparison with a
fully sulfated analogue No. 175 having the following formula:
These results show the importance of the N-acyl groups in the compounds
according to the invention, which make them particularly suitable for use as
medicaments.
are useful in pathologies requiring activation of angiogenesis post-ischaemic
revascularization.
Thus, according to another of its aspects, a subject of the invention is thus
medicaments that comprise a compound of formula (l)/(l') according to the invention, or
a pharmaceutically acceptable salt thereof.
These medicaments find their use in therapy, especially in the treatment of
ischaemia (cardiac ischaemia, arterial ischaemia of the lower limbs), the treatment of
diseases associated with narrowing or obstruction of the arteries or arterites, the
treatment of angina pectoris, the treatment of thromboangitis obliterans, the treatment
of atherosclerosis, the treatment of inhibition of restenosis after angioplasty or
endoarterectomy, the treatment of cicatrization, muscle regeneration treatment,
treatment for the survival of myoblasts, the treatment of peripheral neuropathy, the
treatment of post-operative nerve damage, the treatment of nerve deficiencies such as
Parkinson's disease, Alzheimer's disease, prion disease and neuronal degeneration in
alcoholics, the treatment of dementias, treatment for improving the survival of a
bioartificial pancreas graft in the case of diabetics, treatment for improving the
revascularization of grafts and the survival of grafts, the treatment of retinal
degeneration, the treatment of pigmentary retinitis, the treatment of osteoarthritis, the
treatment of pre-eclampsia or the treatment of vascular lesions and of acute respiratory
distress syndrome, treatment for cartilage repairing, treatment for repairing and
protecting bones, treatment for repairing and protecting hair follicles and for protecting
and regulating hair growth.
Ischaemia is a decrease in arterial circulation in an organ, leading to a decrease
in oxygen concentration in the damaged tissues. In the mechanisms of post-ischaemic
revascularization, two main mechanisms are involved: angiogenesis and arteriogenesis.
Angiogenesis is the process of generating new blood capillaries from pre-existing
vessels. Arteriogenesis contributes towards the development (increase in size and
calibre) of the collateral vessels around the ischaemic or avascular area.
Among the growth factors involved in these revascularization processes, the
FGF family and especially FGF-2 has been the most widely described (Post, M. J.,
Laham, R., Sellke, F. W. & Simons, M. Therapeutic angiogenesis in cardiology using
protein formulations. Cardiovasc. Res. 49, 522-31, 2001). Thus, FGF2 and its receptors
represent very pertinent targets for therapies directed towards inducing angiogenesis
and arteriogenesis processes (Khurana, R. & Simons, M. Insights from angiogenesis
trials using fibroblast growth factor for advanced arteriosclerotic disease. Trends
Cardiovasc. Med. 13, 116-22, 2003).
One of the applications of the compounds of the invention is post-ischaemic
treatment after heart occlusion or occlusion of the peripheral arteries. As regards the
treatment of cardiac ischaemia, one of the most promising clinical tests is a clinical test
in which FGF-2 was sequestered in alginate microspheres in the presence of heparin
(Laham, R. J. et al. Local perivascular delivery of basic fibroblast growth factor in
patients undergoing coronary bypass surgery: results of a phase I randomized, double-
blind, placebo-controlled trial. Circulation 100, 1865-71, 1999). These microspheres
were implanted close to the ischaemic locus in the myocardium. After 90 days, all the
patients treated with FGF2 showed no ischaemic cardiac symptoms. In comparison, in
the control group, three of the seven patients had persistent symptoms at 90 days and
two patients required vascular surgery. Interestingly, the therapeutic benefit was
maintained after 3 years of monitoring. These observations suggest that compounds
that mimic FGF2 may represent a therapy of choice for treating the consequences of
cardiac ischaemia.
Three clinical tests on the injection of FGF2 into the coronary artery were
performed during treatment of narrowing of the coronary arteries (Laham, R. J. et al.
lntracoronary basic fibroblast growth factor (FGF-2) in patients with severe ischemic
heart disease: results of a phase I open-label dose escalation study. J. Am. Coll.
Cardiol. 36, 2132-9, 2000; Simons, M. et al. Pharmacological treatment of coronary
artery disease with recombinant fibroblast growth factor-2: double-blind, randomized,
controlled clinical trial. Circulation 105, 788-93, 2002; Linger, E. F. et al. Effects of a
single intracoronary injection of basic fibroblast growth factor in stable angina pectoris.
Am. J. Cardiol. 85, 1414-9, 2000). The result of these three tests shows that intra-
coronary infusions of FGF2 are well tolerated and significantly improve the condition of
the patients. Thus, the compounds described in the invention may find an application in
the treatment of diseases associated with narrowing of the coronary arteries and
especially in the treatment of angina pectoris.
Diseases of the distal arteries and especially arteritis of the lower limbs are
caused by chronic obstruction of the arterioles that irrigate the extremities. These
pathologies mainly affect the lower limbs. In a phase I clinical trial, patients with
peripheral artery pathologies leading to claudication received injections of FGF2
(Lazarous, D. F. et al., Basic fibroblast growth factor in patients with intermittent
claudication: results of a phase I trial. J. Am. Coll. Cardiol. 36, 1239-44, 2000). In this
context, FGF2 was well tolerated in these patients and the clinical data suggest a
beneficial effect of FGF2 and especially on improving walking. These clinical data
suggest that the compounds of the invention represent a therapeutic tool of choice for
the treatment of diseases associated with obstruction of the distal arteries.
Buerger's disease or thromboangitis obliterans affects the distal vascular
structures and is characterized by distal arteritis of the legs, with pain and ulceration. In
this context, an induction of angiogenesis and of vasculogenesis would represent a
therapy for this pathology. The compounds of the said invention represent a therapy of
choice for thromboangitis obliterans.
Peripheral neuropathy is an axonal or demyelinizing attack of the motor and/or
sensory peripheral nerve which leads to desensitization of the distal limbs. One of the
major secondary complications of diabetes is the chronic development of peripheral
neuropathy. In this context, it has been demonstrated that FGF2 induces axonal
regeneration, which might be a therapy of choice in the treatment of peripheral nerve
lesion and thus in peripheral neuropathy (Basic fibroblast growth factor isoforms
promote axonal elongation and branching of adult sensory neurons in vitro.
Klimaschewski L, Nindl W, Feurle J, Kavakebi P, Kostron H. Neuroscience.
2004;126(2):347-53). By virtue of the agonist activity on the FGF receptors, the
compounds of the said invention would represent a treatment of choice in peripheral
neuropathy in the case of healthy or diabetic patients.
It is clearly established that FGF2 is an activator of nerve cells during
development. Recent results suggest that FGF2 is also a pivotal factor for promoting
the regeneration of neurons in adults (Sapieha PS, Peltier M, Rendahl KG, Manning
WC, Di Polo A., Fibroblast growth factor-2 gene delivery stimulates axon growth by
adult retinal ganglion cells after acute optic nerve injury. Mol. Cell. Neurosci. 2003 Nov.;
24(3):656-72.). By virtue of their agonist activities on the FGF receptors, the compounds
of the said invention would represent a treatment of choice in repairing post-operative
nerve damage, in repairing nerve deficiencies such as Parkinson's disease, Alzheimer's
disease, prion disease and neuronal degeneration in alcoholics or in the case of
dementia.
The proliferation and migration of vascular smooth muscle cells contributes
towards intimal hypertrophy of the arteries and thus plays a predominant role in
atherosclerosis and in restenosis after angioplasty and endoarterectomy. It has been
demonstrated that an angiogenic factor, VEGF, significantly reduces the thickening of
the intima by accelerating re-endothelialization (Van Belle, E., Maillard, L, Tio, F. O. &
Isner, J. M. Accelerated endothelialization by local delivery of recombinant human
vascular endothelial growth factor reduces in-stent intimal formation. Biochem. Biophys.
Res. Commun. 235, 311-6, 1997). Thus, the compounds of the present invention, with
pro-angiogenic activity, may be useful in treatment of atherosclerosis and in inhibiting
restenosis after angioplasty or endoarterectomy.
The vascular network is essential to the development and maintenance of
tissues. By promoting the delivery of nutrients, oxygen and cells, the blood vessels help
to maintain the functional and structural integrity of tissues. In this context, angiogenesis
and vasculogenesis make it possible to preserve and to perfuse tissues after ischaemia.
The angiogenic growth factors such as VEGF and FGF2 thus promote revascularization
for tissue regeneration. The compounds presented in the invention could represent a
treatment of choice in muscle regeneration treatment.
The processes of muscle regeneration on dystrophic or normal muscles depend
on the supply of cytokines and of angiogenic growth factors at the local level (Fibbi, G.,
D'Alessio, S., Pucci, M., Cerletti, M. & Del Rosso, M. Growth factor-dependent
proliferation and invasion of muscle satellite cells require the cell-associated fibrinolytic
system. Biol. Chem. 383, 127-36, 2002). It has been proposed that the FGF system is a
critical system of muscle regeneration and of myoblast survival and proliferation
(Neuhaus, P. et al. Reduced mobility of fibroblast growth factor (FGF)-deficient
myoblasts might contribute to dystrophic changes in the musculature of
FGF2/FGF6/mdx triple-mutant mice. Mol. Cell. Biol. 23, 6037-48, 2003). FGF2 and the
compounds of the said invention could be exploited in order to promote cardiac
regeneration. They would thus improve the perfusion of the myocardium after ischaemia
(Hendel, R. C. et al. Effect of intracoronary recombinant human vascular endothelial
growth factor on myocardial perfusion: evidence for a dose-dependent effect.
Circulation 101, 118-21, 2000) and also the survival and progress of transplanted
myoblasts, especially in Duchenne's muscular dystrophy.
Angiogenesis is an essential phenomenon during cutaneous cicatrization. The
new vessels formed supply the oxygen and nutrients required for tissue repair. In the
case of diabetics, cicatrization is a slow and difficult process presenting angiogenesis
defects. FGFs are among the growth factors that are the most involved in angiogenesis
processes during the cicatrization phase. Certain FGFs are highly overregulated in
dermal cells after a cutaneous injury. On account of their agonist activity on the FGF
receptors, the compounds of the said invention would represent a therapy of choice for
the treatment of cicatrization in healthy or diabetic patients.
Bioartificial pancreas transplantation is a very promising technique for the
treatment of certain types of diabetes. It has been demonstrated, in diabetic rats, that
vascularisation in bioartificial pancreases is much greater when the pancreases are
impregnated with microspheres bearing FGF2 (Sakurai, Tomonori; Satake, Akira, Sumi,
Shoichiro, Inoue, Kazutomo, Nagata, Natsuki, Tabata, Yasuhiko. The Efficient
Prevascularization Induced by Fibroblast Growth Factor 2 With a Collagen-Coated
Device Improves the Cell Survival of a Bioartificial Pancreas. Pancreas. 28(3):e70-e79,
April 2004). This revascularization thus improves the survival of implanted bioartificial
pancreases and consequently the survival of the graft. By virtue of their agonist
activities on the FGF receptors, the compounds of the said invention would represent a
therapy of choice in improving the survival of bioartificial pancreas grafts in diabetics
and more generally in improving the revascularization of grafts and consequently the
survival of the grafts.
Pigmentary retinitis is a pathology involving progressive degeneration of the
retina characterized by degeneration of the photoreceptors and obliteration of the retinal
vessels. Lahdenranta et al. (An anti-angiogenic state in mice and humans with retinal
photoreceptor cell degeneration. Proc. Natl. Acad. Sci. USA 98, 10368-73, 2001) have
proposed that angiogenic growth factors regulate the neural coordination and the
associated vascularization of the retina by simultaneously functioning as photoreceptor
survival factors and as endothelial cell regulators. In this context, the intra-vitreal
injection of FGF2 retards the degeneration of the photoreceptors by acting on retinal
survival and retinal angiogenesis (Faktorovich, E. G., Steinberg, R. H., Yasumura, D.,
Matthes, M. T. & LaVail, M. M. Basic fibroblast growth factor and local injury protect
photoreceptors from light damage in the rat. J. Neurosci. 12, 3554-67,1992). These
observations demonstrate the interest of the compounds described in the invention as a
therapy in retinal degeneration and especially in pigmentary retinitis.
In the field of osteoarthritis, many studies have been performed for restoring
destroyed articular cartilage. In this context, it has been reported that the proliferation
and differentiation of chondrocytes were stimulated by FGF2 in vitro (Kato Y,
Gospodarowicz D. Sulfated proteoglycan synthesis by confluent cultures of rabbit costal
chondrocytes grown in the presence of fibroblast growth factor. J. Cell Biol. 1985 Feb.;
100(2):477-85). Furthermore, Cuevas et al. have shown that FGF2 induces cartilage
repair in vivo (Cuevas P, Burgos J, Baird A. Basic fibroblast growth factor (FGF)
promotes cartilage repair in vivo. Biochem. Biophys. Res. Commun. 1988 Oct.
31;156(2):611-8). Takafuji et al. have also shown that FGF2 implants significantly
improve temporo-mandibular cartilage in rabbits suffering from osteoarthritis (Takafuji
H, Suzuki T, Okubo Y, Fujimura K, Bessho K Regeneration of articular cartilage defects
in the temporomandibular joint of rabbits by fibroblast growth factor-2: a pilot study. Int.
J. Oral Maxillofac. Surg. 2007 Oct.; 36(10):934-7). These observations demonstrate the
interest the compounds described in the invention as a therapy in treatment of
osteoarthritis and cartilage repair.
In the field of bone repair, one of the essential needs is to find agents that
stimulate bone formation. Among the main growth factors, it is established that the
systemic administration of FGF2 facilitates bone repair (Acceleration of fracture healing
in nonhuman primates by fibroblast growth factor-2. Kawaguchi H, Nakamura K, Tabata
Y, Ikada Y, Aoyama I, Anzai J, Nakamura T, Hiyama Y, Tamura M. J. Clin. Endocrinol.
Metab. 2001 Feb.; 86(2), 875-880). The local application of FGF2 in gelatin matrices
accelerates bone repair in primates, suggesting the clinical utility of FGF2 in the
treatment of fractures. By virtue of their agonist properties for the FGF receptors, the
compounds of the said invention would represent a treatment of choice in bone repair.
Pre-eclampsia is a pathology of the placenta associated with a vascularization
defect (Sherer, D. M. & Abulafia, O. Angiogenesis during implantation, and placental
and early embryonic development. Placenta 22, 1-13, 2001). These vascularization
defects are thought to be due to an angiogenesis defect and lead to placental
disruptions that may result in the death of the foetus. The compounds of the invention
may be a treatment of choice for overcoming an angiogenesis defect in pre-eclamptic
placentas.
In addition to angiogenesis-inducing effects, growth factors such as VEGF or
FGF2 protect endothelial cells against intrinsic and extrinsic apoptosis inducers. The
intrinsic signalling pathway is activated by the mitochondria in response to a stress such
as deprivation or DNA damage, whereas the extrinsic signalling pathway is induced by
the binding of pro-apoptotic factors such as TNF-a or Fas. It is now clearly described
that VEGF and FGF2 are two factors of endothelial cell survival (Role of Raf in Vascular
Protection from Distinct Apoptotic Stimuli: A Alavi, J.D. Hood, R. Frausto, D. G.
Stupack, DA Cheresh: Science 4 July 2003: Vol. 301. No. 5629, pp. 94-96). Acute
respiratory distress syndrome (ARDS) is characterized by cardiovascular and
neuropsychiatric problems. In the context of the cardiovascular problems, the patients
present major vascular lesions and especially a high induction of apoptosis of
endothelial cells. Recently, Hamacher et al. have demonstrated that the
bronchoalveolar lavage fluids of patients suffering from ARDS showed pro-apoptotic
activity against lung microvascular endothelial cells (Tumor necrosis factor-alpha and
angiostatin are mediators of endothelial cytotoxicity in bronchoalveolar lavages of
patients with acute respiratory distress syndrome. Am. J. Respir. Crit. Care Med. 2002
Sep. 1;166(5):651-6: Hamacher J., Lucas R., Lijnen H.R., Buschke S., Dunant Y.,
Wendel A., Grau G.E., Suter P.M., Ricou B.). By virtue of their activity on endothelial
cell survival, the products of the invention might be a treatment of choice in the vascular
improvement of patients suffering from vascular lesions and especially patients
suffering from ARDS.
The endogenous overregulation of FGF7 (or KGF) and of FGF18 appears to be
an important mechanism for promoting the proliferation, migration and protection of hair
follicles in pathological cases or after a tumoral treatment (Comprehensive Analysis of
FGF and FGFR Expression in Skin: FGF18 Is Highly Expressed in Hair Follicles and
Capable of Inducing Anagen from Telogen Stage Hair Follicles. Mitsuko Kawano, Akiko
Komi-Kuramochi, Masahiro Asada, Masashi Suzuki, Junko Oki, Ju Jiang and Toru
Imamura). By virtue of their agonist activity on the FGF receptors, the compounds of
the said invention might be a treatment of choice for repairing and protecting hair
follicles and for protecting and regulating hair growth.
According to another of its aspects, the present invention relates to
pharmaceutical compositions comprising, as active principle, a compound according to
the invention. These pharmaceutical compositions contain an effective dose of at least
one compound according to the invention, or a pharmaceutically acceptable salt of the
said compound, and also at least one pharmaceutically acceptable excipient. The said
excipients are chosen, according to the pharmaceutical form and the desired mode of
administration, from the usual excipients known to those skilled in the art.
In the pharmaceutical compositions of the present invention for oral, sublingual,
subcutaneous, intramuscular, intravenous, topical, local, intratracheal, intranasal,
transdermal or rectal administration, the active principle of formula (l)/(l') above or the
salt thereof may be administered in a unit administration form, as a mixture with
standard pharmaceutical excipients, to man and animals for the prevention or treatment
of the above disorders or diseases.
The appropriate unit administration forms include oral forms such as tablets, soft
or hard gel capsules, powders, granules and oral solutions or suspensions, sublingual,
buccal, intratracheal, intraocular, intranasal or inhalation administration forms, topical,
transdermal, subcutaneous, intramuscular or intravenous administration forms, rectal
administration forms and implants. For topical application, the compounds according to
the invention may be used in creams, gels, ointments or lotions.
The injectable administration forms are particularly advantageous, conventionally
comprising the active compound dissolved in water for injection, in the presence of
sodium chloride. The unit dose of active compound should be suited to the desired
therapeutic effect; it may be, for example, between 0.1 and 100 mg of active principle.
According to another of its aspects, the present invention also relates to a
method for treating the pathologies indicated above, which comprises the administration
to patients of an effective dose of a compound according to the invention or a
pharmaceutically acceptable salt thereof.
CLAIMS
1. Octasaccharide compounds corresponding to formula (I):

in which:
- the wavy line denotes a bond located either below or above the plane of
pyranose ring of the glucosamine unit,
- R1 represents a group O-alkyl, in which the said alkyl group comprises fror
to 16 carbon atoms and is optionally substituted with one or more groups, which may
identical or different, chosen from aryl and cycloalkyl groups,
- R2 represents either a group OSO3" or a hydroxyl group,
- R3 represents an alkyl, cycloalkyl or alkyl-cycloalkyl group, and
- R4 represents a disaccharide of formula (II):

in which:
- R3 is as defined above,
- R5 represents either a group OSO3" or a hydroxyl group, and
- R6 represents a disaccharide of formula (III):

in which:
- R3 is as defined above,
- R7 represents either a group OSO3" or a hydroxyl group, and
- R8 represents a disaccharide of formula (IV):
in which:
- R3 is as defined above,
- R9 represents either a group OSO3- or a hydroxyl group,
in acid form or in the form of any pharmaceutically acceptable salt thereof.
2. Compounds according to Claim 1, in which R1 represents:
- either a group O-alkyl, in which the said alkyl group comprises from 5 to 16
carbon atoms,
- or a group O-alkyl, in which the said alkyl group comprises from 1 to 8 carbon
atoms and is substituted with 1 or 2 groups, which may be identical or different, chosen
from aryl and cycloalkyl groups.
3. Compounds according to Claim 1 or Claim 2, in which R1 represents a group
O-alkyl, in which the said alkyl group comprises from 5 to 12 carbon atoms.
4. Compounds according to any one of Claims 1 to 3, in which R1 represents a
group O-alkyl, in which the said alkyl group comprises from 3 to 6 carbon atoms and is
substituted with 1 or 2 groups, which may be identical or different, chosen from phenyl
and cycloalkyl groups.
5. Compounds according to any one of Claims 1 to 4, in which R3 represents
either an alkyl group comprising from 2 to 6 carbon atoms, or a cycloalkyl group.
6. Compounds according to any one of Claims 1 to 5, in which at least one of the
groups R2, R5, R7 and Rg represents a hydroxyl group.
7. Compounds according to any one of Claims 1 to 6, in which at least one of the
groups R2, R5, R7 and R9 represents a hydroxyl group and at least one of the groups R2,
R5, R7 and R9 represents a group OSO3-.
8. Compounds according to any one of Claims 1 to 7, characterized in that they
correspond to formula (I'), in which R1, R2, R3, R5.R7 and R9 are as defined in any one
of Claims 1 to 7:
9. Compounds according to any one of Claims 1 to 8, chosen from the following
compounds:
- Pentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium
sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-
(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-acetamido-2-
deoxy-6-O-sodium sulfonato-ß-D-glucopyranoside (No. 1);
- Pentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
deoxy-2-(1-oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium
2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1 -oxobutyl)amino-6-
O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopyranosyluronate)-(1?4)-(2-deoxy-2-(1 -oxobutyl)amino-6-O-sodium sulfonato-a-D-
glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-
2-deoxy-2-(1-oxobutyl)amino-6-O-sodium sulfonato-ß-D-glucopyranoside (No. 2);
- Pentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4))-(2-
deoxy-2-(1-oxohexyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium
2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1 -oxohexyl)amino-6-
O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopyranosyluronate)-(1?4)-(2-deoxy-2-(1-oxohexyl)amino-6-O-sodium sulfonato-a-D-
g!ucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-
2-deoxy-2-(1-oxohexyl)amino-6-O-sodium sulfonato-ß-D-glucopyranoside (No. 3);
- Pentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
[(cyclopenty!carbonyl)amino]-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-
(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-[(cyclopentyl-
carbonyl)amino]-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-
O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4))-(2-[(cyclopentylcarbonyl)amino]-2-
deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-
a-L-idopyranosyluronate)-(1?4)-2-[(cyclopentylcarbonyl)amino]-2-deoxy-6-O-sodium
sulfonato-p--D-glucopyranoside (No. 4);
- Undecyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium
sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-
(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-acetamido-2-
deoxy-6-O-sodium sulfonato-ß-D-glucopyranoside (No. 5);
- 5-phenylpentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-
(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium
sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-
(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-acetamido-2-
deoxy-6-O-sodium sulfonato-ß-D-glucopyranoside (No. 6);
- Methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-[(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium
sulfonato-a-D-glucopyranosyl)]2-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronate)-(1?4)-2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranoside (No. 7);
- Methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium
sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronate)-(1?4)-(2-acetamido-2-deoxy-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium
sulfonato-a-L-idopyranosyluronate)-(1?4)-2-acetamido-2-deoxy-6-O-sodium sulfonato-
a-D-glucopyranoside (No. 8);
- Methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodiurn 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4))-(2-acetamido-2-deoxy-6-O-sodium
sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyl-
uronate)-(1?4)-(2-acetamido-2-deoxy-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium
sulfonato-a-L-idopyranosyluronate)-(1?4)-2-acetamido-2-deoxy-6-O-sodium sulfonato-
a-D-glucopyranoside (No. 9);
- Methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-gluco-
pyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4))-2-acetamido-2-deoxy-a-D-gluco-
pyranoside (No. 10);
- Methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-6-O-sodium sulfonato-a-D-gluco-
pyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-acetamido-2-deoxy-6-O-sodium
sulfonato-a-D-glucopyranoside (No. 11);
- Methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-a-D-gluco-
pyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-acetamido-2-deoxy-a-D-gluco-
pyranoside (No. 12);
- Methyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-acetamido-2-deoxy-a-D-gluco-
pyranosyI)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-
acetamido-2-deoxy-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-
sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-acetamido-2-deoxy-6-O-sodium
sulfonato-a-D-glucopyranoside (No. 13);
- 5-phenylpentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-
(2-deoxy-2-(1-oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-
(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1-
oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium
sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1-oxobutyl)amino-a-D-gluco-
pyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-
deoxy-2-(1-oxobutyl)amino-ß-D-glucopyranoside (No. 14);
- 2-propylpentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-
(2-deoxy-2-(1-oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-
(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1-
oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium
sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1-oxobutyl)amino-a-D-gluco-
pyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-
deoxy-2-(1-oxobutyl)amino-ß-D-glucopyranoside (No. 15);
- 3-cyclohexylpropyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-
(1?4)-(2-deoxy-2-acetamido-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium
2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-acetamido-6-O-
sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopyranosyluronate)-(1?4)-(2-deoxy-2-acetamido-a-D-glucopyranosyl)-(1?4)-(sodium
2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-deoxy-2-acetamido-|3--D-gluco-
pyranoside (No. 16);
- 3-cyclohexylpropyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-
(1?4)-(2-deoxy-2-(1-oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-
(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1-
oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium
sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1-oxobutyl)amino-a-D-gluco-
pyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-
deoxy-2-(1-oxobutyl)amino-ß-D-glucopyranoside (No. 17);
- 3,3-diphenylpropyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-
(1?4)-(2-deoxy-2-(1 -oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-
(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1 -
oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium
sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(1-oxobutyl)amino-a-D-gluco-
pyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-2-
deoxy-2-(1-oxobutyl)amino-ß-D-glucopyranoside (No. 18); and
- 5-phenylpentyl (sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-
(2-deoxy-2-(3-methyl-1-oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-
(1?4)-(sodium 2-O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(3-
methyl-1-oxobutyl)amino-6-O-sodium sulfonato-a-D-glucopyranosyl)-(1?4)-(sodium 2-
O-sodium sulfonato-a-L-idopyranosyluronate)-(1?4)-(2-deoxy-2-(3-methyl-1-
oxobutyl)amino-o-a-D-glucopyranosyl)-(1?4)-(sodium 2-O-sodium sulfonato-a-L-
idopyranosyluronate)-(1?4)-2-deoxy-2-(3-methyl-1-oxobutyl)amino-(3--D-gluco-
pyranoside (No. 19).
10. Medicament, characterized in that it comprises a compound according to any
one of Claims 1 to 9, or a pharmaceutically acceptable salt thereof.
11. Pharmaceutical composition, characterized in that it comprises a compound
according to any one of Claims 1 to 9, or a pharmaceutically acceptable salt thereof,
and also at least one pharmaceutically acceptable excipient.
12. Compound according to any one of Claims 1 to 9, for its use in the treatment
of pathologies requiring activation of the FGF receptors.
13. Compound according to Claim 12, for its use in the treatment of pathologies
requiring angiogenesis activation and post-ischaemic revascularization.
14. Compound according to Claim 12 or Claim 13, for its use in the treatment of
ischaemia, such as cardiac ischaemia and arterial ischaemia of the lower limbs, the
treatment of diseases associated with narrowing or obstruction of the arteries or
arterites, the treatment of angina pectoris, the treatment of thromboangitis obliterans,
the treatment of atherosclerosis, the treatment of inhibition of restenosis after
angioplasty or endoarterectomy, the treatment of cicatrization, muscle regeneration
treatment, treatment for the survival of myoblasts, the treatment of peripheral
neuropathy, the treatment of post-operative nerve damage, the treatment of nerve
deficiencies such as Parkinson's disease, Alzheimer's disease, prion disease and
neuronal degeneration in alcoholics, the treatment of dementias, treatment for
improving the survival of a bioartificial pancreas graft in the case of diabetics, treatment
for improving the revascularization of grafts and the survival of grafts, the treatment of
retinal degeneration, the treatment of pigmentary retinitis, the treatment of
osteoarthritis, the treatment of pre-eclampsia or the treatment of vascular lesions and of
acute respiratory distress syndrome, treatment for repairing cartilage, treatment for
repairing and protecting bones, treatment for repairing and protecting hair follicles and
for protecting and regulating hair growth.
15. Compound of formula (V), in which Alk represents an alkyl group, R1 is as
defined in any one of Claims 1 to 4, and Pg, Pg' and Pg", which may be identical or
different, represent protecting groups:
16. Compound according to Claim 15, in which Pg, Pg' and Pg" represent,
respectively, benzyl, tBDPS and acetyl groups, Alk represents a methyl group and R1 is
as defined in any one of Claims 1 to 4.
17. Compound according to Claim 15 or 16, in which Pg, Pg' and Pg" represent,
respectively, benzyl, tBDPS and acetyl groups, Alk represents a methyl group and R1 is
chosen from the following groups: -O-methyl, -O-(CH2)5-phenyl, -O-CH2-CH(C3H7)2, -O-
(CH2)3-C6H11 and -O-(CH2)2-CH(phenyl)2.
18. Compound of formula (VI), in which Alk represents an alkyl group and Pg,
Pg' and Pg", which may be identical or different, represent protecting groups:
19. Compound according to Claim 18, in which Alk represents a methyl group
and Pg, Pg' and Pg" represent, respectively, benzyl, levulinyl and acetyl groups.

Abstract:

The invention relates to FGF receptor-activating N-acyl octasaccharides having Formula (I), wherein: R1 is an O-
alkyl group optionally replaced by one or more aryl or cycloalkyl groups, R2 is an OSO3- or hydroxyl group, R3 is an alkyl,
cycloalkyl, or alky-cycloalkyl group, R4 is a disaccharide having Formula (II), R6 is a disaccharide having Formula (III), and R8 is
a disaccharide having Formula (IV), where R5, R7, and R9 have OSO3 or hydroxyl groups. The invention further relates to the
preparation of said octasaccharides and to the therapeutic use thereof.