Title of Invention

AZASUGAR DERIVATIVES, HEPARANASE INHIBITORS AND PHARMACEUTICAL COMPOSITIONS THEREOF

Abstract The invention concerns heparanase inhibiting compounds of general formula (I) wherein R represents a hydrogen atom, an hydroxyl radical, a -OSO3- radical, a -O-(C1-C5 alkyl radical or an -O- aralky) radival; Z represents a COO radical or a hydroxy) radical; X represents -OH or a saccharide unit of formula A, Y represents H, C1-C3 alkyl or a saccharide unit of formula D; in fire form or in the form of pharmaceutically aceeptable salts formed with a base or an acid as well as in the form of solvates or hydrates. The derivatives of the invention are useful as medicines.
Full Text The subject of the invention is azasugar derivatives, inhibitors of heparanases, their
preparation, compositions containing them and their
therapeutic application..
Heparanases are enzymes of the
endoglucuronidase type which have as substrate
polysaccharides of the heparin/heparan sulphate (HS)
family. Type I and II heparanases are known (McKenzie
et al., Biochem. Biophys. Res. Commun. (2000), Vol.
276, p. 1170-1177) . They hydrolyse specifically β-l->4
type bonds between a saccharide unit of the
D-glucuronic acid type and a saccharide unit of the
D-glucosamine type and release HS fragments of about 10
to 20 saccharide units (Pikas, D.S. et al., J. Biol.
Chem., (1998), Vol. 2 73, p. 18770-18777.). Heparanases
break down in the same manner the polysaccharide chains
of heparan sulphate proteoglycans (HSPGs) (Vlokavsky
and Friedmann, J. Clin Invest., (2001), Vol. 108, p.
341-347). HSPGs consist of a core protein to which
linear HS chains are covalently attached (Kjellen et
al., Annu. Rev. Biochem., (1991), Vol. 60, p. 443-475).
HSPGs are ubiquitous macromolecules. Like HSs, HSPGs
are present at the surface of numerous cell types and in the extracellular matrix (ECM) (Kjellen et al.,

(1991), ibid.} Bernfield et al., Annu Rev. Biochem.,
(1999), Vol. 68, p. 729-777; David et al., FASEB J.,
(1993), Vol. 7, p. 1023-1030; Lozzo et al., Annu. Rev.
Biochem. (1998), Vol. 67, p. 609-652). The ECM, a major
component of the connective tissues of vertebrates and
invertebrates, occupies the extracellular environment.
It envelops the organs and surrounds the endothelia, in
particular the capillary endothelia (Wight et al.,
Arteriosclerosis, (1989), Vol. 9, p. 1-20), thus
playing a role of maintenance and of barrier for
protection of the organs and endothelia (McKenzie et
al.r Biochem. J.; 2003; Vol. 373, p. 423-435). The ECM
is also a key modulator involved in various cellular
mechanisms, in particular cell differentiation and
repair (Folkman et al., Adv. Exp. Med. Biol., (1992),
Vol. 313, p. 355-364).
Heparanase inhibiting compounds have been
described in the prior art. For example, International
Patent Application WO 02/0600374 describes benz-1,3-
azole derivatives, International Patent Application
WO 03/074516 relates to phthalimidecarboxylic acid and
benzoxazole derivatives, International Patent
Application WO 04/013132 describes furanthiazole
derivatives or International Patent Application
WO 04/046123 describes benzoxazole, benzothiazole and
benzimidazole derivatives.
The synthesis of short-chain (2 units)

azasugar derivatives in which the nitrogen atom
replaces the oxygen atom at the 5-position has been
described in Takahashi et al., Chem. Lett., (1994),
Vol. 11, p. 2119; Takahashi et al., Tetrahedron,
(2001), Vol. 57, p. 6915-6926). However, their
activities in vivo have not been identified.
Azasugar derivatives with a single unit, of
the following formula:

have already been described (Patent US 6,583,158;
Ichikawa et al., J. Amer. Chem. Soc., (1998), Vol. 12 0,
p. 3007) .
A need still exists to find and to develop
products having good activity in vitro and in vivo.
It has now been found, surprisingly, that
synthetic azasugar derivatives exhibit good activity as
heparanase inhibitors. The present invention therefore
relates to novel azasugar derivatives which are
heparanase inhibitors. These novel compounds exhibit
good heparanase inhibiting activity.
The subject of the invention relates to
compounds corresponding to the general formula (I):


in which:
R represents a hydrogen atom, a hydroxyl radical, an
-OSO3- radical, an -O- (C1-C5) alkyl radical or an
O-aralkyl radical;
Z represents a COO- radical or a hydroxyl radical;
X represents a hydroxyl radical or a saccharide unit of
formula A:

in which:
- R1 represents an oxygen atom, allowing A to bind
to the azasugar unit or to another saccharide
unit,
- R2 represents an -NH2 radical, an -NHCO (C1-C5) -
alkyl radical, an -NHCOaryl radical, an -NHSO3-
radical, a hydroxyl radical, an -O-(C1-C5)alkyl
radical, an -O-aralkyl radical or an -0SO3"
radical,
- R3 represents a hydroxyl radical, an -0SO3"
radical, an -O- (C1-C5) alkyl radical or an
-O-aralkyl radical,
- R4 represents a hydroxyl radical, an -0SO3"


radical, an -O-(C1-C5) alkyl radical, an O-aralkyl
radical or a saccharide unit of formula B:

in which:
- R6 represents an oxygen atom, allowing B to
bind to another saccharide unit of formula A,
- R7 and R8 have the same definition as R3 as
defined above,
- R9 represents a hydroxyl radical, an -0SO3-
radical, an -O-(C1-C5)alkyl radical, an
-O-aralkyl radical or a saccharide unit of
formula A ,as defined above,
- R5 has the same definition as R3 as defined
above;
Y represents a hydrogen atom, a (C1-C5)alkyl radical or
a saccharide unit of formula D

in which:
- R10, R12 and R13 have the same definitions as R5,
R3 and R2 respectively as defined above,
- R11 represents:

. a (C1-C3)alkylene radical allowing D to
attach to the azasugar unit, or
. an oxygen atom allowing D to attach to
another saccharide unit,
- R14 represents an. -O-(C1-C5) alkyl radical or an
-O-E radical in which E represents a radical of
the following formula:

in which:
- R15 represents an -O- (C1-C5) alkyl radical, an
-O-aralkyl radical or a saccharide unit of
formula D in which R11 represents an oxygen
atom,
- R16 and R17 have the same definition as R3 as
defined above,
provided, however, that when X and R each,represent a
hydroxyl radical, Y does not represent a hydrogen atom,
and it being understood that the number of saccharide
units of which the compound of formula (I) is composed
is between 1 and 10,
in free form or in the form of salts formed with a
pharmaceutically acceptable base or acid, and in the
form of solvates or hydrates.
According to one of its preferred aspects,

the invention relates to the compounds of general
formula (I):
in which:
R represents a hydroxyl radical;
Z represents a COO- radical or a hydroxyl radical;
X represents a hydroxyl radical or a saccharide unit of
formula A:
in which:
- R1 represents an oxygen atom,
- R2 represents an -NHCOCH3 radical, an -NHSO3"
radical, an -0SO3" radical,
- R3 represents a hydroxyl radical or an
-O-(C1-C5) alkyl radical,
- R4 represents a hydroxyl radical, an -O-aralkyl
radical or a saccharide unit of formula B:

in which:

- R6 represents an oxygen atom,
- R7 represents an -OSO3- radical,
- R8 represents a hydroxyl radical, an
-O- (C1-C5) alkyl radical or an -O-aralkyl
radical,
- R9 represents an -OSO3- radical, an
-O-aralkyl radical, an -O-(C1-C5)alkyl radical
or a saccharide unit of formula A as defined
above,
- R5 represents an -0SO3- radical;
Y represents a hydrogen atom or a saccharide unit of
formula D:
in which:
- R10 has the same definition as R5 as defined
above,
- R12 represents a hydroxyl- radical or an -0SO3"
radical,
- R13 represents an -NHSO3- radical,
- R11 represents a methylene radical linked to an
azasugar unit or an oxygen atom linked to E,
- R14 an -OCH3 radical or a radical of formula -O-E
in which E represents a radical of formula:


in which:
- R15 represents a D unit in which R11
represents an oxygen atom allowing E to be
linked to D,
- R16 represents an -OSO3~ radical,
- R17 represents a hydroxyl radical,
it being understood that the number of saccharide units
of which the compound of formula (I) is composed is
between 2 and 10,
in free form or in the form of salts with a
pharmaceutically acceptable base or acid, and in the
form of solvates or hydrates.
Particularly preferred compounds are
compounds of formula I in which Y is a hydrogen atom.
The invention encompasses azasugar
derivatives in their acid form or in the form of any
one of their pharmaceutically acceptable salts. In the
acid form, the -COO- and -SO3~ functional groups are in
the -COOH and -SO3H forms respectively.
According to one of its particularly
preferred aspects, the present invention relates to the
following compounds:
• (2,4-di-O-sodium sulphonato-α-L-idopyranosyl-

uronate of sodium)-(1-4)-(2-acetamido-2-deoxy-6-
O-sodium sulphonato-α-D-glucopyranosyl)-(1-4)-
(2-O-sodium sulphonato-α-L-idopyranosyluronate
of sodium)-(1-4)-(2-acetamido-2-deoxy-6-O-sodium
sulphonato-α-D-glucopyranosyl)-(1-4)-
(5-(hydroxy)-4-oxypiperidine-3-carboxylate of
sodium (3S, 4R, 5R)) (compound No. 20)
(2,4-di-O-sodium sulphonato-α-L-idopyranosyl-
uronate of sodium)-(1-4)-(2-N-sodium sulphonato-
2-deoxy-6-O-sodium sulphonato-α-D-gluco-
pyranosyl)-(1-4)-(2-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(2-N-sodium
sulphonato-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranosyl)-(1-4)-(5-(hydroxy)-4-oxy-
piperidine-3-carboxylate of sodium (3S, 4R, 5R))
(compound No. 2 7)
(3-O-methyl-2,4-di-O-sodium sulphonato-ce-L-
idopyranosyluronate of sodium)-(1-4)-(3-O-
methyl-2,6-di-O-sodium sulphonato-α-D-gluco-
pyranosyl)-(1-4)-(3-O-methyl-2-O-sodium
sulphonato-α-L-idopyranosyluronate of sodium)-
(1-4) - (3-O-methyl-2, 6-di-O-sodium sulphonato-α;-
D-glucopyranosyl)-(1-4)-(5-(hydroxy)-4-oxy-
piperidine-3-carbpxylate of sodium (3S, 4R, 5R))
(compound No. 47)
(2,4-di-O-sodium sulphonato-α-L-idopyranosyl-
uronate of sodium)-(1-4)-(2,6)-di-O-sodium

sulphonato-α-D-glucopyranosyl)-(1-4)-(2-O-sodium
sulphonato-α-L-idopyranosyluronate of sodium)-
(1-4)-(2,6-di-O-sodium sulphonato-α-D-gluco-
pyranosyl)-(1-4)-(5-(hydroxy)-4-oxypiperidine-3-
carboxylate of sodium (3S, 4R, 5R)) (compound
No. 69)
(4-O-propyl-2-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(2,6-di-O-
sodium sulphonato-α-D-glucopyranosyl)-(1-4)-(2-
O-sodium sulphonato-α-L-idopyranosyluronate of
sodium) - (1-4) - (2 , 6-di-O-sodium sulphonato-α-D-
glucopyranosyl)-(1-4)-(5-(hydroxy)-4-
oxypiperidine-3-carboxylate of sodium (3S, 4R,
5R)) (compound No. 74)
(2,4-di-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(2-
acetamido-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranosyl) - (1-4) - (2-O-sodium sulphonato-o;-
L-idopyranosyluronate of sodium)-(1-4)-(2-
acetamido-2-deoxy-6-O-sodium sulphonato-ce-D-
glucopyranosyl)-(1-4)-(3-(hydroxy)-5-
hydroxymethyl-4-oxypiperidine (3R, 4R, 5R))
(compound No. 123)
(4-O-phenylpropyl-2-O-sodium sulphonato-ce-L-
idopyranosyluronate of sodium)-(1-4)-(2-
acetamido-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranosyl)-(1-4)-(2-O-sodium sulphonato-α-

L-idopyranosyluronate of sodium)-(1-4)-(2-
acetamido-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranosyl)-(1-4)-(5-(hydroxy)-4-oxy-
piperidine-3-carboxylate of sodium (3S, 4R, 5R))
(compound No. 124).
In the context of the present invention:
a (C1-C5)alkyl radical represents a linear or
branched saturated aliphatic radical which may
contain from 1 to 5 carbon atoms. By way of
examples, there may be mentioned methyl, ethyl,
propyl, isopropyl, butyl, isobutyl and tert-butyl
radicals and the like,
an -O-aralkyl radical represents an alkyl group as
defined above but substituted with an aromatic
radical which may itself carry substituents. By
way of example, there may be mentioned
p-methoxybenzyl, phenylmethyl, phenylethyl,
phenylpropyl and the like,
a (C1-C3)alkylene radical represents a divalent
alkyl group containing from 1 to 3 carbon atoms.
The expression pharmaceutically acceptable
salt is understood to mean azasugar derivatives of the
invention, an azasugar derivative in which one or more
of the -COO- or/and -SO3- functional groups are
ionically linked to a pharmaceutically acceptable
cation. The preferred salts according to the invention
are those whose cation is chosen from alkali metal

cations and still more preferably those whose cation is
Na+ or K+.
In its principle, the method for preparing
the compounds according to the invention uses mono-,
di- or oligosaccharide parent synthons prepared as
previously reported in the literature and chosen taking
into account in particular the orthogonality of the
protecting groups. Reference may be made in particular
to EP 300099, EP 529715, EP 621282 and EP 649854 and to
C. van Boeckel, M. Petitou, Angew. Chem. Int. Ed.
Engl., (1993), Vol. 32, p. 1671-1690. These synthons
are then coupled to each other so as to provide a
completely protected equivalent of a compound according
to the invention. This protected equivalent is then
converted to a compound according to the invention
using methods well known to the person skilled in the
art.
As the compounds of the invention
additionally contain an azasugar (or substituted
piperidine) unit, their synthesis requires the
preparation of a precursor of this unit carrying
protecting groups compatible with subsequent couplings
to mono-, di- or oligosaccharides. The azasugar
precursors are prepared according to methods described
in the literature. Reference may be made in particular
to the book "Iminosugars as Glycosidase Inhibitors",
AE Stiitz, Wiley-VCH, 1999.

When the synthons necessary for the assembly
of the chain are available, couplings of these synthons
with each other are carried out. In the coupling
reactions mentioned, a "donor" synthon, activated on
its anomeric carbon, reacts with an "acceptor" synthon,
possessing at least one free hydroxyl.
The present invention relates to a method for
the preparation of the compounds of formula (I),
characterized in that: in a first step, a completely
protected, equivalent of the desired compound (I) is
prepared; in a second step, the negatively charged
groups (carboxylates, sulphonates) and the free
hydroxyls are introduced and/or unmasked.
The synthesis of the precursor is carried out
according to reactions well known to persons skilled in
the art using in particular the methods of saccharide
synthesis (G.J. Boons, Tetrahedron, (1996), Vol. 52, p.
1095-1121; WO 98/03554 and WO 99/36443) according to
which a glycosidic bond donor oligosaccharide is
coupled with a glycosidic bond acceptor 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 desired compound of
formula (I) is obtained. The structure of the desired
final compound determines the nature of the chemical
entities used in the various steps of the synthesis, so
as to control the stereochemistry and the

regioselectivity, according to rules well known to
persons skilled in the art.
The compounds according to the invention are
obtained from their completely protected polysaccharide
precursors using in general the following sequence of
reactions:
release and substitution of the functional groups
which have to be converted to N- and/or O-
sulphonate groups;
release of the other functional groups and
optional assembly of the fragments with each
other, for example by a reductive amination
reaction.
The release of the carboxylic acid functional groups
may be carried out at various stages of this method.
The compounds of the invention may naturally
be prepared using various strategies known to persons
skilled in the art for saccharide and organic
synthesis.
The method described above is the preferred
method of the invention. However, the compounds of
formula (I) may be prepared by other methods well known
in sugar chemistry which are 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 in G.J. Boons, Tetrahedron,
(1996), Vol. 52, p. 1095-1121.

The protecting groups used in the method for
preparing the compounds (I) are those commonly used in
sugar chemistry, for example in Protective Groups in
Organic Synthesis, TW Greene, PGM Wuts, John Wiley &
Sons, New York, 1999.
The protecting groups are advantageously
chosen for example from acetyl, halomethyl, benzoyl,
levulinyl, benzyl, substituted benzyl, optionally
substituted trityl, carbamate, tetrahydropyranyl,
allyl, pentenyl, tert-butyldimethyl silyl (tBDMS) or
trimethylsilylethyl groups.
The activating groups are those
conventionally used in sugar chemistry according to for
example G.J. Boons, Tetrahedron, (1996), Vol. 52, p.
1095-1121. These activating groups are chosen for
example from imidates, thioglycosides, pentenyl-
glycosides, xanthates, phosphites or halides.
The method described above makes it possible
to obtain compounds of the invention in the form of
salts. To obtain the corresponding acids, the compounds
of the invention in the form of salts are brought into
contact with a cation exchange resin,in acid form.
The compounds of the invention in the form of
acids may then be neutralized with a base in order to
obtain the desired salt. For the preparation of the
salts of the compounds of formula (I), it is possible
to use any inorganic or organic base which gives

pharmaceutically acceptable salts with the compounds of
formula (I). Sodium, potassium, calcium or magnesium
hydroxide is preferably used as base. The sodium and
calcium salts of the compounds of formula (I) are the
preferred salts.
The compounds according to the invention have
been the subject of biochemical and pharmacological
studies. The following nonlimiting tests illustrate the
present invention.
The following terms are defined:
PET: polyethylene terephthalate,
AM: acetoxymethyl,
DMEM: Dulbecco's modified Eagle's medium,
EDTA: ethylenediaminetetraacetic acid,
Tris: tris(hydroxymethyl)aminomethane,
AT: antithrombin III,
nkat: nanokatal = enzymatic unit of measurement (given
by the manufacturer) representing the quantity of
substrate catalysed per unit of time.
1. Evaluation of the activity of the heparanase
inhibitors in an enzymatic system (determination of the
IC50 values of the compounds according to the invention)
The heparanase activity is demonstrated by
its. capacity to degrade fondaparinux. The concentration
of fondaparinux is determined by means of its anti-
factor Xa activity.
A. Materials and methods

The heparanase is produced by Sanofi-
Synthelabo (Labege, France).
The reagents for assaying factor Xa are
marketed by Chromogenix (Montpellier, France).
Increasing concentrations of a compound
according to the invention, an inhibitor of heparanases
(varying dilutions: from 1 nM to 10 µM) are mixed at a
fixed heparanase concentration (for each batch,
preliminary experiments make it possible to determine
the enzymatic activity which is sufficient for the
degradation of 0.45 µg/ml of added fondaparinux) . After
5 minutes at 3 7°C, the mixture is exposed to
fondaparinux and left for 1 hour at 37°C. The reaction
is stopped by heating at 95°C for 5 minutes. The
residual fondaparinux concentration is finally -measured
by adding factor Xa and its specific chromogenic
substrate (Ref. S2222).
The various mixtures are produced according
to the following procedure:
a) Reaction mixture
50 µl of sodium acetate buffer (0.2 M,
pH 4.2) are mixed with 50 µl of fondaparinux
(0.45 µg/ml) and 59 µ1 of a heparanase solution. The
mixture is incubated for 1 hour at 3 7°C and then for
5 minutes at 95°C. 100 µl of the reaction mixture are
then mixed with 50 µl of 50 mM Tris buffer containing
175 mM NaCl, 75 mM EDTA, pH 14. The pH thus passes from

4.2 to 7.
"The anti-factor Xa activity of fondaparinux
is measured in the following manner:
b) Assay of the anti-factor Xa activity of
fondaparinux
100 µl of the solution obtained in step a)
are mixed with 100 µl of AT (0.5 µ/ml) . The mixture is
kept for 2 minutes at 37°C and 100 µl of factor Xa
(7 nkat/ml) are then added. The mixture is kept for
2 minutes at 3 7°C and 10 0 µl of chromogenic substrate
(Ref.: S2222) (1 mM) are then added. The mixture is
kept for 2 minutes at 37°C and 100 µl of acetic acid
(5 0%) are then added.
The optical density is read at 405 nm.
A percentage inhibition is determined
relative to control without inhibitor. A curve of the
percentage inhibition makes it possible to calculate an
IC50.
B. Results
The compounds according to the present
invention have IC50 values of between 10 nM and 10 /zM.
For example, compound No. 27 has an IC50 of
11+4 nM (mean + SD, done on two assays).
2. Effect of the heparanase inhibitors on the invasion
of HT1080 tumour cells
The effect of the compounds of formula (I),
inhibitors of heparanases, was tested in vitro on the

invasion of HT1080 tumour cells.
A. Materials and methods:
a) Cell culture:
The cells derived from human fibrosarcomas,
HT1080 (ATCC CCL-121) are cultured in a DMEM medium
(Ref.: GIBCO 1196O-044) containing 5% Foetal Calf
Serum, glutamine (2 mM) (Ref.: GIBCO 2503O-024), sodium
pyruvate (1 mM) (Ref.: GIBCO 1136O-039) and
nonessential amino acids (1X) (Ref.: GIBCO 1114O-035),
on collagen coated flasks (Becton Dickinson 75 cm2; Ref.
354523), to 50 to 80% confluence.
b) Cell invasion test
The measurements of invasion of the HT1080
cells are carried out on a Becton Dickinson falcon HTS
Fluoroblock Multiwell Insert System kit in 24-well
plates (Ref.: 351158). These measurement chambers give
the cells the conditions which make it possible to
evaluate their invasive properties in vitro.
The kit is composed of a plate combined with
culture inserts containing a PET membrane pierced with
8 micron pores on which a uniform Matrigel Matrix
(Becton Dickinson; Ref. 354230) layer is deposited.
Matrigel is a soluble basal membrane
extracted from the EHS (Engelbreth-Holm-Swarm) tumour
which, by virtue of its composition, forms, upon
solidifying, a structure equivalent to a basal
membrane.

The Matrigel layer blocks the pores of the
membrane, thus blocking the migration of the
noninvasive cells across the membrane. By contrast, the
invasive (tumour or nontumour) cells will be capable of
becoming detached and of invading the Matrigel layer
before migrating across the membrane.
The quantification of the cell migration is
carried out by labelling with calcein AM (Molecular
Probes C-3100). The fluorescent signal emitted is
measured with the aid of a Perkin Elmer Wallac VICTOR 3
reader and may be directly correlated with the number
of cells which have invaded the Matrigel gel. By
comparing them with controls made in the same
experiment as the products studied (response in the
presence of 0% and 5% foetal calf serum), it is
possible to determine a percentage inhibition of cell
invasion in the presence of the products.
B. Results
A series of independent determinations
(varying from 2 to 4) made it possible to show that at
a concentration of between 1 nM and 10 jiM, the
compounds according to the invention inhibit cell
invasion on average by a percentage of between 8 and
60%.
For example, a series of four independent
determinations made it possible to show that 10 /iM of
compound No. 2 0 inhibit cell invasion on average by a

percentage equal to 40.3 ± 8.3% (mean ± standard
deviation).
Furthermore, compound No. 2 0 has a dose-
dependent effect on cell invasion.
Indeed:
at a concentration of 0.3 µM, compound No. 20
inhibits cell invasion by 26 ± 3%,
at a concentration of 1 µM, compound No. 2 0
inhibits cell invasion by 53 + 11%.
These results demonstrate an increase in the
inhibition of cell invasion as a function of the dose
of compound No. 20.
The compounds of formula (I) according to the
present invention therefore exhibit good affinity for
heparanases and exhibit a heparanase inhibiting effect.
It has been demonstrated in animals and in
humans that the increase in the secretion of
heparanases and cancerous progression are correlated
(Goldschmit et al., PNAS, (2002), Vol. 99(15) , p.
10031-10036). For example, a high heparanase level has
been detected in the serum of animals having metastatic
tumours (Vlodavsky et al., Isr. J. Med. Sci. (1988),
Vol. 24(9-10), p. 464-470) or in the urine of patients
suffering from cancer who have developed numerous
metastases (Vlodavsky et al., Curr. Biol., (1997), Vol.
7(1), p.43-51). Tumour biopsies have shown the same
correlation (Vlodavsky et al., Isr. J. Med. Sci.,

(1988), Vol. 24(9-10), p. 464-470). A correlation
therefore exists between the increase in secretion of
heparanases and the metastatic potential of tumour
cells (Vlodavsky et al. , Invasion Metastasis, (1994),
Vol.14, p. 29O-302; Nature Medicine, (1999), Vol. 5, p.
793-802) .
Heparanases, which are secreted by tumour
cells, degrade the HSPGs and HS, which are major
components of the ECM. The ECM thus perforated allows
the tumour and metastatic cells to circulate and also
allows the invasion of newly-formed blood vessels
(angiogenesis) (Suzanne A. Eccles, Nat. Med., (1999),
Vol. 5 (7),p. 793-809). Angiogenesis is a process for
generating new capillary vessels from preexisting
vessels or by mobilization and differentiation of bone
marrow cells. Thus, both an uncontrolled proliferation
of the endothelial cells and a mobilization of
angioblasts from the bone marrow are observed in the
new vascularization processes of tumours.
Thus, heparanases represent relevant targets
for therapies aimed at inhibiting the processes of
invasion of cancer cells and of metastasization, on the
one hand, and of angiogenesis, on the other. The
expression cancer (or carcinoma) is understood to mean
any malignant cell growth of the epithelium, present in
the skin but also and especially in the wall of the
organs and the appearance of metastatic tumour cells

such as melanomas, mesothelioma, lymphoma, leukaemia,
fibrosarcoma, rhabdomyosarcoma, mastocytoma, but also
carcinomas affecting a tissue such as the colon, the
rectum, the prostate, the lungs, the breasts, the
pancreas, the intestine, the kidneys, the ovaries, the
uterus, the cervix, the bladder, the liver and the
stomach. The carcinomas infiltrate the adjacent tissues
and spread (metastasize) to other distant organs, for
example the liver, the lungs, the brain or the bones. A
compound possessing a heparanase inhibiting activity,
such as the compounds of the invention, may therefore
be useful for the treatment of such cancers (Fang J et
al., Proc. Natl. Acad. Sci. USA, (2000), Vol. 97(8), p.
3884-3889; Kondraganti et al., Cancer Res., (2000),
Vol. 60 (24) , p. 6851-6855), and in particular
colorectal, prostate, lung, breast, pancreatic, kidney,
bladder and ovarian cancer.
The p75 receptor, a receptor for molecules of
the neurotrophin (NT) family, has been identified as
being a representative marker of the phenomenon of
metastasization in the brain. It has furthermore been
demonstrated that the secretion of NT involves an
increase in the secretion of heparanases (Marchetti D.
et al. J. Cell. Biochem., (2004), Vol. 91(1), p. 206-
215). Thus, a compound possessing a heparanase
inhibiting activity, such as the compounds of the
invention, may therefore be useful for reducing

metastasization in the central nervous system by
inhibiting the action of the activation of the p75
receptor (Marchetti D. et al., Pathol. Oncol. Res.,
(2003) Vol. 9(3), p. 147-158 and Epub, (2003), Review;
Walch ET. et al., Int. J. Cancer, (1999), Vol. 82 (1),
p. 112-120; Menter DG. et al., Invasion Metastasis,
(1994-95), Vol. 14 (1-6) , p. 372-384; Marchetti D. et
al., Int. J. Cancer, (1993), Vol. 55 (4) , p. 692-699).
The activity of heparanases on the HSPGs of
the ECM also appears to be correlated with the onset of
inflammatory.and autoimmune reactions (Vlodavsky et
al., Invasion metastasis, (1994), Vol.. 4, p. 29O-302;
Goldschmit et al., Med. Sci. (2002), Vol. 99 (15), p.
10031-10036). The interaction of the platelets, the
granulocytes, the B and T lymphocytes, the macrophages
and the mastocytes with the ECM is associated with the
degradation of the HSs by the heparanases (Vlodavsky et
al., Invasion Metastasis, (1992), Vol. 12, p. 112-127)..
Thus, a compound such as the compounds of the
invention, possessing a heparanase inhibiting activity,
may therefore be useful for the treatment of
inflammatory diseases, in particular chronic
inflammatory diseases such as rheumatoid arthritis or
IBD (Inflammatory Bowel Disease), comprising two forms
of chronic inflammatory bowel diseases: UC (ulcerative
colitis) and Crohn's disease (CD) or autoimmune
diseases.

Other studies suggest that heparanases could
play a role in the treatment of cardiovascular diseases
{Journal of Pharmacological Sciences, (2004), _94, No.
Supplement 1, pp. 160P, print.; and Meeting Info.: 77th
Annual Meeting of the Japanese Pharmacological Society,
Osaka, Japan. March 08-10, 2004. Japanese
Pharmacological Society; and Miller, Heather Ann, Diss.
Abstr. Int., (1984), B 2003, 64 (5) ; Demir et al. , Clin.
Appl. Thromb. Hemost., (2001), Vol. 7(2), p. 131-140),
such as post-angioplasty restenosis, diseases linked to
the vascular complications of diabetes such as diabetic
retinopathies, atherosclerosis (Atherosclerosis,
(1999), Vol. 145, p. 97-10.6; J. Clin. Invest., (1997),
Vol. 10 0, p. 867-874) or thromboembolic diseases and
arterial thromboses. Thus, a heparanase inhibiting
compound according to the invention may represent a
therapy of choice in these pathologies.
Moreover, heparanases are known to be
involved in certain cases of renal insufficiency where
the renal filtration and reabsorption functions may be
impaired (FASEB J, (2004), Vol. 18, p. 252-263).. Thus,
a heparanase inhibiting compound according to the
invention may represent a therapy of choice in such
pathologies.
The HSPGs of the ECM also appear to play a
role as major regulators of cell growth and activation
via the modulation of growth factors, in particular of

FGFs (Fibroblast Growth Factors). For example, the
activity of heparanases involves the release of growth
factors such as the FGFs, which stimulate in particular
angiogenesis, and promotes tumour progression (Bashkin
et al., Biochemistry, (1989), Vol. 28, p. 1737-1743).
Thus, heparanases represent relevant targets for the
treatment of diseases in which the FGFs are involved.
In general, the FGFs are involved to a great
extent via autocrine, paracrine or juxtacrine
secretions in the phenomena of deregulation of the
stimulation of the growth of cancer cells. Furthermore,
the FGFs initiate tumour angiogenesis which plays a
predominant role both on tumour growth and also on the
phenomena of metastasization.
Angiogenesis is a process for generating new
capillary vessels from preexisting vessels or by
mobilization and differentiation of bone marrow cells.
Thus, both uncontrolled proliferation of the
endothelial cells and mobilization of angioblasts from
the bone marrow are observed in the processes of
neovascularization of tumours. It has been shown in
vitro and in vivo that several growth factors stimulate
endothelial proliferation, and in particular FGF-1 or
a-FGF and FGF-2 or b-FGF.
a-FGF and b-FGF play for example an important
role in the growth and the maintenance of prostate
cells (Doll JA, et al., Prostate, (2001), Vol. 305, p.

49-293.
Several research studies show the presence of
a-FGF and b-FGF and of their receptors (FGFRs) both in
human breast tumour lines (in particular MCF7) and in
tumour biopsies.
Glioma cells produce a-FGF and b-FGF in vitro
and in vivo and possess various FGF receptors at their
surface.
More recently, the potential role of
proangiogenic agents, and in particular of FGFs, in
leukaemias and lymphomas has been documented (Thomas DA
et al., Acta Haematol, (2001), Vol. 207, p. 106-190).
A correlation exists between the process of
bone marrow angiogenesis and extramedullar diseases in
CMLs (chronic myelomonocytic leukaemia).
The proliferation and migration of vascular
smooth muscle cells contribute to the intimal
hypertrophy of the arteries and thus plays a
predominant role in atherosclerosis and in restenosis
following angioplasty and endarterectomy. Studies in
vivo show a local production of a-FGF and b-FGF after
lesion of the carotid by balloon injury.
Vascular disorders due to diabetes are
characterized by an impairment of the vascular
reactivity and of the blood flow, a hyperpermeability,
an increased proliferative response and an increase in
matrix protein deposits. More precisely, a-FGF and

b-FGF are present in the preretinal membranes of
patients with diabetic retinopathies, in the membranes
of the underlying capillaries and in the vitreous
humour of patients suffering from proliferative
retinopathies.
Rheumatoid arthritis (RA) is a chronic
disease with unknown etiology. While it affects
numerous organs, the most severe form of RA is a
progressive synovial inflammation of the joints
resulting in destruction. Angiogenesis appears to
greatly affect the progression of this pathology. Thus,
a-FGF and b-FGF have been detected in the synovial
tissue and in the joint fluid of patients suffering
from RA, indicating that this growth factor is involved
in the initiation and/or the progression of this
pathology. In AIA models (adjuvant-induced model of
arthritis) in rats, it has been shown that the
overexpression of b-FGF increases the severity of the
disease whereas an anti b-FGF neutralizing antibody
blocks the progression of RA (Yamashita et al. , J.
Immunol., (2002), Vol. 57, pl 168-450; Manabe et al.,
Rheumatol, (1999), Vol. 20, p. 38-714).
Angiogenesis and inflammation are also major
phenomena which occur in the processes involved in
osteoarthritis leading to destruction of the joint
accompanied by pain. Angiogenesis may also play a role
in chondrocytic hypertrophy and ossification, thus

contributing to the modifications of the joint (Bonnet
CS et al, Rheumatology. (1) 7-16 2005) .
IBDs (inflammatory bowel diseases) comprise
two forms of chronic inflammatory bowel diseases: UC
(ulcerative colitis) and Crohn's disease (CD). IBDs are
characterized by an immune dysfunction resulting in an
inappropriate production of inflammatory cytokines
inducing the establishment of a local microvascular
system. The consequence of this angiogenesis of
inflammatory origin is an intestinal ischaemia induced
by vasoconstriction. High circulating and local b-FGF
levels have been measured in patients suffering from
these pathologies (Kanazawa et al., American Journal of
Gastroenterology, (2001), Vol. 28, p. 96-822; Thorn et
al., Scandinavian Journal of Gastroenterology, (2000),
Vol. 12, p. 35-408) .
A compound possessing a heparanase inhibiting
activity, such as the compounds of the invention, may
therefore be useful for the treatment of diseases
linked to an up regulation of the FGFs and/or of their
receptors.
By virtue of their low toxicity and their
pharmacological and biological properties, the
compounds of the present invention find application in
the treatment of any carcinoma having a high degree of
vascularization (lung, breast, prostate, oesophagus) or
inducing metastases (colon, stomach, melanoma) or

sensitive to a-FGF or to b-FGF in an autocrine manner
or finally in pathologies of the lymphoma and leukaemia
type. These compounds represent a therapy of choice
either alone or in combination with a suitable
chemotherapy or radiotherapy or in combination with a
treatment with antiangiogenic agents. The compounds
according to the invention also find application in the
treatment of cardiovascular diseases such as
atherosclerosis, post-angioplasty restenosis, in the
treatment of diseases linked to the complications which
appear following the fitting of endovascular prostheses
and/or aortocoronary bypass surgery or vascular
complications of diabetes such as diabetic
retinopathies. The compounds of the invention also find
application in the treatment of chronic inflammatory diseases such as rheumatoid arthritis or IBDs.
The products according to the invention also
find application in the treatment of macular
degeneration. A major characteristic of the loss of
vision in adults is the neovascularization and the
subsequent haemorrhages which cause considerable
functional disorders in the eye and which result in
early blindness. Recently, the study of the mechanisms
involved in the phenomena of ocular neovascularization
has made it possible to demonstrate the involvement of

the proangiogenic factor in these pathologies.
The compounds of the invention may also be

used in combination with one or more anticancer
treatments, such as surgical treatments, radiotherapy
or in combination with compounds which block
angiogenesis. For example, the compounds of the
invention may be used alone or in combination with
another active ingredient such as cisplatine,
cyclophosphamide, methotrexate, 5-fluorouracil,
paclitaxel, docetaxel, vincristine, vinblastine,
vinorelbine, doxorubicin, tamoxifen, toremifene,
megestrol acetate, anastrozole, goserelin, capecitabine
and raloxifene or molecules having an antiangiogenic
activity, for the treatment of cancer.
According to another of its aspects, the
subject of the present invention is therefore a
pharmaceutical composition containing, as active
ingredient, a compound of formula (I) in free form or
in the form of salts formed with a pharmaceutically. acceptable base or acid, according to the invention,
optionally in combination with one or more inert and
appropriate excipients.
The said excipients are chosen according to
the desired pharmaceutical dosage form and mode of
administration: oral, sublingual, subcutaneous,
intramuscular, intravenous, transdermal, transmucosal,
local or rectal.
In each dosage unit, the active ingredient is
present in quantities appropriate for the daily doses

envisaged in order to obtain the desired prophylactic
or therapeutic effect. Each dosage unit may contain
from 0.1 to 100 mg of active ingredient, preferably 0.5
to 50 mg.
The pharmaceutical compositions of the
invention may be intended for oral, sublingual,
subcutaneous, intramuscular, intravenous,
intratracheal, topical, intranasal, transdermal,
rectal, intraocular and vaginal administration. The
unit forms for administration may be for example
tablets, gelatin capsules, granules, powders, oral or
injectable solution or suspensions, patches, injector
pens and suppositories. For local administration,
ointments, creams, lotions, eyedrops, gels, sprays and
oil may be envisaged.
The said unit forms contain doses which allow
a daily administration of 1 to 100 mg of active
ingredient per kg of body weight, according to the
galenic form used.
To prepare tablets, a pharmaceutical vehicle
which may be composed of diluents, such as, for
example, lactose, microcrystalline cellulose and
starch, ,,-and formulation adjuvants, such as binders
(polyvinylpyrrolidone, hydroxypropylmethylcellulose,
and the like), glidants such as silica, and lubricants
such as magnesium stearate, stearic acid, glyceryl
tribehenate and sodium stearylfumarate, is added to the

active ingredient, micronized or not. Wetting agents or
surfactants such as sodium lauryl sulphate may also be
added.
The production techniques may be direct
compression, dry granulation, wet granulation or hot-
melt.
The tablets may be uncoated, coated for
example with sucrose, or coated with various polymers
or other appropriate materials. They may be designed to
allow rapid, delayed or prolonged release of the active.
ingredient by virtue of the polymer matrices or the
specific polymers used in the coating.
To prepare gelatin capsules, the active
ingredient is mixed with dry pharmaceutical vehicles
(simple mixture, dry or wet granulation, or hot-melt),
liquid pharmaceutical vehicles or semi-solid
pharmaceutical vehicles.
The gelatin capsules may be hard or soft, film-coated or not, so as to have a rapid, prolonged or
delayed activity (for example for an enteric form).
A composition in syrup or elixir form or for
administration in the form of droplets may contain the
active ingredient together with a sweetener, preferably
a calorie-free sweetener, methylparaben or
propylparaben as antiseptic, a saliva modifier and a
colouring.
The water-dispersible powders and granules

may contain the active ingredient in the form of a
mixture with dispersing agents or wetting agents, or
dispersing agents such as polyvinylpyrrolidone, and
with sweeteners and flavour corrigents.
For rectal administration, suppositories
prepared with binders which melt at the rectal
temperature, for example cocoa butter or polyethylene
glycols, are used.
For parenteral administration, aqueous
suspensions, isotonic salt solutions or injectable
sterile solutions containing pharmacologically
compatible dispersing agents and/or wetting agents, for
example propylene glycol or butylene glycol, are used.
The active ingredient may also be formulated
in the form of microcapsules, optionally with one or
more carriers or additives, or with a polymer matrix or
with a cyclodextrin (patches, prolonged-release forms).
The compositions for local administration
according to the invention comprise a medium compatible
with the skin. They may be provided in particular in
the form of aqueous, alcoholic or aqueous-alcoholic
solutions, gels, water-in-oil or oil-in-water emulsions
having the appearance of a cream or a gel,
microemulsions, aerosols, or even in the form of
vesicular dispersions containing ionic and/or nonionic
lipids. These galenic forms are prepared according to
the customary methods of the fields considered.

The active ingredient may also be formulated
in the form of microcapsules, optionally with one or
more carriers or additives, or with a polymer matrix or
with a cyclodextrin (patches, prolonged-release forms).
The local compositions according to the
invention comprise a medium compatible with the skin.
They may be provided in particular in the form of
aqueous, alcoholic or aqueous-alcoholic solutions,
gels, water-in-oil or oil-in-water emulsions having the appearance of a cream or of a gel, microemulsions,
aerosols, or in the form of vesicular dispersions
containing ionic and/or nonionic lipids. These galenic
forms are prepared according to the customary methods
of the fields considered.
The examples which follow, given without
limitation, illustrate the preparation of compounds
according to the present invention.
Abbreviations used in the text which follows,:
Ac = acetyl
All = allyl
Bn = benzyl
Me = methyl
Ph = phenyl
PMB = (4-methoxy)benzyl
PMBBr = (3-bromo-4-methoxy)benzyl
Z = benzyloxycarbonyl
TLC = thin-layer chromatography

The examples which follow illustrate the
preparation of compounds according to the invention,
without limiting it. Before embarking on the
preparation of these various examples, there is
described below the preparation of compounds
(PREPARATION) useful for the production of compounds of
the invention or for other preparations.
For the preparations and examples which
follow, the following experimental, methods are used:
METHOD 1
Oxidation of the primary alcohols to an acid, and then
conversion to a benzyl ester
TEMPO (0.02 molar equivalent) and a saturated
aqueous sodium hydrogen carbonate solution (4 1/mol)
are added to a solution of compound to be oxidized
(1 molar equivalent) in tetrahydrofuran (THF)
(3.5 1/mol). After cooling to 0°C, Bromodan (2 molar
equivalents) is added dropwise over 2 0 min. After 3 h
of magnetic stirring, the reaction mixture is
concentrated and the residue is dried by repeated
evaporation of dimethylformamide (DMF) (4.95 1/mol).
The crude compound thus obtained is used as it is in
the next step.
A solution of the preceding compound in
dimethylformamide (13.1 1/mol) is treated at room
temperature (l-15h) with benzyl bromide (10 molar
equivalents), and potassium hydrogen carbonate (5 molar

equivalents). The reaction mixture is concentrated and
then the residue is dissolved in ethyl acetate
(35 1/mol), washed with water, dried (sodium sulphate)
and concentrated. Column chromatography gives the
expected benzyl ester.
METHOD 2
Coupling to imidates catalysed by tert-butyldimethyl-
silyl triflate
A solution of tejrt-butyldimethylsilyl
triflate in dichloromethane (0.1M, 0.15 mol per mole of
imidate) is added, under argon, at -20°C, to a solution
of the imidate and of the glycosyl acceptor in a
dichloromethane/diethyl ether mixture (1:2,
22.5-45 1/mol) in the presence of 4 A molecular sieves.
After 1O-45 minutes (TLC), solid sodium hydrogen
carbonate is added. The solution is filtered, washed
with water, dried and evaporated to dryness. METHOD 3
Method for saponification of the esters
Hydrogen peroxide.(H2O2) at 3 0% (7.16 1/mol
ester) and an aqueous 0.7N lithium hydroxide solution
(2.3 mol per mole of ester) are successively added, at
-5°C, to a solution of compound to be saponified in
tetrahydrofuran (160 1/mol). After stirring for 1 h at
-5°C, the reaction medium is placed for 4 h at 0°C and
then stirred at room temperature until the esters have
been consumed. The crude reaction product is then

optionally purified on an LH-20 column.
METHOD 4
Sulphonation
Triethylamine/sulphur trioxide complex (5 mol
per hydroxyl functional group) is added to a solution
in dimethylformamide (90 1/mol) of the compound to be
sulphated. After 12 to 22 hours at 55°C, methanol or an
aqueous sodium hydrogen carbonate solution is added at
0°C, and after stirring for 0.5-24 h at room
temperature, the reaction medium is purified with the
aid of an LH-2 0 column, or of two Sephadex" G-25 columns
(eluted successively with a 0.2M aqueous sodium
chloride solution, and then with water). The fractions
containing the product are then concentrated under a
high vacuum to give the desired product.
METHOD 5
Hydrogenolysis of the benzyl ethers and/or of the
benzyl esters
A solution of the compound in glacial acetic
acid/water/tert-butanol mixture is kept stirring for
6-16 h (TLC) under a hydrogen atmosphere (3-15 bar) in
the presence of 5 or 10% palladium on carbon
(equivalent to 0.7-3 times the mass of the compound).
After filtration, the solution is deposited at the top
of a Sephadex G-25 column, eluted with 0.2M sodium
chloride. The fractions containing the product are
concentrated and desalted using the same column eluted

lwith water. The final compound is obtained after
freeze-drying.
Preparations useful for producing the
compounds according to the invention are described
below.

Step 1.a: Preparation of (3R,4R,5R)-1-benzyl-3-
(benzyloxy)-5-(hydroxymethyl)piperidin-4-ol (No. 2)
The synthesis of compound 1 is described in
T.M. Jespersen and M. Bois, Tetrahedron (1994) 5J3 (47) ,
13449-13460 and in Patent.No. US 5,844,102. The
synthesis of compound 2 is described in Patent
WO 98/50359.
To a solution of compound 1 (10.8 g,
42.8 mmol) in methanol (590 ml) are successively added
sodium cyanoborohydride (5.38 g, 2 molar equivalents),
followed by acetic acid (7.4 ml, 3 molar equivalents)

at -10°C and a benzylamine solution (5.1 ml,
1.1 equivalent), in methanol (100 ml). After returning
to room temperature, the reaction mixture is heated at
50°C for 2 h. After returning to room temperature, a 2%
sodium hydrogen carbonate solution (85 ml) is added.
The methanol is concentrated under vacuum and then the
residue is diluted with dichloromethane and the organic
phase is washed with water and then with an aqueous
sodium chloride solution, dried (Na2SO4) and then
concentrated under vacuum. The residue is used directly
in the next step without purification.
Step l.b: Preparation of (3R,4R,5R)-methyl [4-(acetyl-
oxy)-1-benzyl-5-(benzyloxy)piperidin-3-yl]acetate
(No. 3)
Triethylamine (13.5 ml, 2.25 molar
equivalents), 4-(dimethylamino)pyridine (DMAP) (7.84 g,
1.5 equivalents and acetic anhydride (61 ml, 15 molar
equivalents) are successively added to a solution of
the crude compound 2 (10.8 g) obtained in step l.a in
dichloromethane (345 ml). The temperature is kept at
0°C for 10 min and then the reaction medium is placed
at room temperature for 16 h. The reaction mixture is
then concentrated under vacuum and the residue purified
on silica gel to give compound 3 (7.65 g, 43%,
2 steps).
XH NMR (CDC13) δ7.36-7.18 (m, 10H, Ar) , 4.6O-4.42 (dd,
2H, OCH2Ph) , 2.02, 1.99 (2s, 6H, 2CH3CO) .

Step l.c: Preparation of (3R, 4R, 5R) -benzyl 4-(acetyl-'
oxy)-3-acetyloxymethyl)-5-(benzyl oxy)piperidine-1-
carboxylate. (No. 4)
Benzyloxycarbonyl chloride (2.4 ml, 3 molar
equivalents) is added under argon, at -10°C, to a
solution of compound 3 (2.31 g, 5.6 mmol) obtained in
step l.b in tetrahydrofuran (28 ml), and then the
reaction medium is left stirring at room temperature
for 18 h. The reaction mixture is then concentrated
under vacuum and the residue is purified on silica gel
(1:9 diethyl, ether - diisopropyl ether) to give
compound 4 (2.14 g, 84%).
Mass spectrum (ESI) m/z 478.3 [ (M + Na)+] .
Step l.d: Preparation of (3R,4R,5R)-benzyl 3-(benzyl-
oxy)-4-hydroxy-5-(hydroxymethyl)piperdine-1-carboxylate
(No. 5)
A solution of 0.84M lithium hydroxide
monohydrate (25 ml, 5 molar equivalents) is added, at
G°C,.to a solution of compound 4 (1.9 g, 4.2 mmol)
obtained in step l.c, in dioxane (25 ml). The reaction
medium is kept at 0°C for 5 minutes and is then placed
at room temperature for 3 0 min. After neutralizing with
hydrochloric acid (HC1:3N), the reaction medium is
diluted in dichloromethane, washed with water, dried
(Na2SO4) , filtered and concentrated. The residue is
purified on silica gel (3:1 ethyl acetate-cyclohexane)
to give compound 5 (1.59 g, 90%)

Mass spectrum (ESI) m/z 394.4 [ (M+Na)+] .
Step l.e: Preparation of (3S,4R,5R)-dibenzyl 5-
(benzyloxy)-4-hydroxypiperidine-1,3-dicarboxylate
(No. 6)
Compound 5 (3.18 g, 8.6 mmol) obtained in
step l.d is treated according to METHOD 1 to give
compound 6 (2.92 g, 71%).
Mass spectrum (ESI) m/z 476.5 [ (M+Na)+] .
PREPARATION 2
Synthesis of (benzyl 2,4-di-O-acetyl-3-O-benzyl-a;-L-
idopyranosyluronate)-(1-4)-(6-O-acetyl-2-azido-3-O-
benzyl-2-deoxy-or, /S-D-glucopyranose trichloro-
acetimidate) (No. 11)

Step 2.a: Preparation of (benzyl 2-Q-acetyl-3-O-benzyl-
q-L-idopyranosyluronate)-(1-4)-(1,6-anhydro-2-azido-3-
O-benzyl-2-deoxy-ff-D-glucopyranose) (No. 8)
Compound 7 in 2'-O-acetylated form (18.0 g,
31.48 mmol), prepared in the same manner as the 2'-O-
benzoylated compound described in Y. Ichikawa et al. ,
Tetrahedron Lett. (1986) 27 (5) 611-614, is treated

according to METHOD 1 to give, after purification on
silica gel (3:7 ethyl acetate-cyclohexane), compound
8 (1.6.4 g, 77%) .
Mass spectrum (ESI) m/z 698.3 [ (M+Na)+]
Step 2.b: Preparation of (benzyl 2,4-di-O-acetyl-3-Q-
benzyl-α-L-idopyranosyluronate)-(1-4)-(1,6-di-O-acetyl-
2-azido-3-O-benzyl-2-deoxy-α, β-D-glucopyranose) (No . 9)
Trifluoroacetic acid (TFA) (4.7 ml, 11 molar
equivalents) is added, at 0°C,.to a solution of
compound 8 (3.74 g, 5.54 mmol) obtained in step 2.a, in
acetic anhydride (52 ml, 100 molar equivalents). After
returning to room temperature, the reaction mixture is
stirred for 16 h and is then concentrated, coevaporated
with toluene and purified on silica gel (4:1 toluene-
ethyl acetate) to give compound 9 (4.33 g, 95%).
Mass spectrum (ESI) m/z 842.2 [ (M+Na)+] .
Step 2.c: Preparation of (benzyl 2,4-di-O-acetyl-3-O-
benzyl-g-L-idopyranosyluronate)-(1-4)-(6-O-acetyl-2-
azido-3-Q-benzyl-2-deoxy-α, β-D-glucopyranose), (No. 10)
Benzylamine (BnNH2) (22 ml, 38 molar
equivalents) is added, at 0°C, to a solution of
compound 9 (4.3: g, 5.24 mmol) obtained in step 2.b, in
diethyl ether (210 ml). After stirring for 4.5 h at
room temperature, the medium is acidified with IN HC1
and is then extracted with ethyl acetate, dried
(Na2SO4) , concentrated and purified on silica gel (35:65
ethyl acetate-cyclohexane) to give 10 (3.4 g, 83%).

Mass spectrum (ESI) m/z 800.2 [ (M+Na)+] .
Step 2.d: Preparation of (benzyl 2,4-di-O-acetyl-3-O-
benzyl-α-L-idopyranosyluronate)-(1-4)-(6-O-acetyl-2-
azido-3 -O-benzyl-2-deoxy-α, /3-D-glucopyranose
trichloroacetimidate) (No. 11)
Caesium carbonate (Cs2CO3) (2.26 g, 1.6 molar
equivalents) and then trichloroacetonitrile (CC13CN)
(1.74 ml, 5.0 molar equivalents) are added, under
argon, to a solution of compound 10 (3.38 g, 4.35 mmol)
obtained in step 2.c, in dichloromethane (82 ml) . After
stirring for 1.5 h, the reaction mixture is filtered
and then concentrated. The residue is purified on
silica gel (3:7 ethyl acetate-cyclohexane) to give 11
(2.96 g, 74%) .
1H NMR (CDC13) δ6.43 (d, H-lce GlcI), 5.64 (d, H-1β
GlcI) , 5.17 (d, IdoUAII).
PREPARATION 3:
Synthesis of (3-O-benzyl-2,4-di-O-sodium sulphonato-α-
L-idopyranosyluronate of sodium)-(1-4)-(2-acetamido-3-
O-benzyl-2-deoxy-6-O-sodium sulphonato-or-D-
glucopyranosyl)-(1-4)-(3-O-benzyl-2-O-sodium
sulphonato-α-L-idopyranosyluronate of sodium)-(1-4)-(2-
acetamido-3-O-benzyl-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranosyl)-(1-4)-(5-(benzyloxy)-4-oxypiperidine-1-
carboxylate of benzyl-3-carboxylate of sodium
(3S,4R,5R)) (No. 19)


Step 3 .a: Preparation of (benzyl 2 , 4-di-'O-acetyl-3-O-
benzyl-g-L-idopyranosyluronate) - (1-4) - (6-O-ac.etyl-2-
azido-3-O-benzyl-2-deoxy-α-D-glucopyranosyl) - (1-4) -
(benzyl 2-O-acetyl-3-O-benzyl-α-L-idopyranosyluronate) -
(1-4) - (1, 6-anhydro-2-azido-3-O-benzyl-2-deoxy-β-D-
glucopyranose) (No. 12)
Compound 11(455 mg, 0.50 mmol) obtained in
step 2.d and compound 8 (675 mg, 1 mmol) obtained in

step 2. a are treated according to method 2 to give,
after purification, compound 12 (385 mg, 54%).
Mass spectrum (ESI) m/z 1457.6 [ (M+Na)+] .
Step 3..b: Preparation of (benzyl 2,4-di-O-'acetyl-3-O-
benzyl-α-L-idopyranosyluronate)-(1-4)-(6-O-acetyl-2-
azido-3-O-benzyl-2-deoxy-α-D-glucopyranosyluronate-(1-
4)-(benzyl 2-O-acetyl-3-O-benzyl-α-L-
idopyranosyluronate)- (1-4)-(1,6-di-O-acetyl-2-azido-3-
O-benzyl-2-deoxy-a?, )6-D-glucopyranose) (No. 13)
Compound 12 (365 mg, 0.254 mmol) obtained in
step 3.a is treated as for the synthesis of compound 9
(step 2.b) to give, after purification on silica gel
(1:1 Et2O-diisopropyl ether), 13 (376 mg, 97%).
Mass spectrum (ESI) m/z 1560.7 [ (M+Na)+] .
Step 3.c: Preparation of (benzyl 2,4-di-O-acetyl-3-O-
benzyl-α-L-idopyranosyluronate)-(1-4)-6-O-acetyl-2-
azido-3-O-benzyl-2-dedxy-α-Drglucopyranosyl)-(1-4)-
(benzyl 2-O-acetyl-3-O-benzyl-α-L-idopyranosyluronate) -
(1-4) - (6-O-acetyl-2-azido-3-O-benzyl-2'-deoxy-α, β-D-
glucopyranose) (No. 14)
Compound 13 (3 64 mg, 0.23 7 mmol) obtained in
step 3.b is treated as for the synthesis of compound 10
(step 2.c) to give, after purification on silica gel
(7:3 Et2O-diisopropyl ether), compound 14. (310 mg, 87%).
Mass spectrum (ESI) m/z 1518.8 [(M+Na)+] .
Step 3.d: Preparation of (benzyl 2,4-di-O-acetyl-3-O-
benzyl-α-L-idopyranosyluronate) - (1-4) -,(6-O-acetyl-2-


azido-3-O-benzyl-2-deoxy-α-D-glucopyranosyl)-(1-4)-
(benzyl .2-O-acetyl-3-Q-benzyl-α-L-idopyranosyluronate) -
(1-4) - (6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-α,β-D-
glucopyranose trichloroacetimidate) (No. 15)
Compound 14 (279 mg, 0.187 ramol) obtained in
step 3.c is treated as for the synthesis of compound 11
(step 2.d) to give, after purification on silica gel
(1:1 Et2O-diisopropyl ether), 15 (230 mg, 75%).
Mass spectrum (ESI) m/z 1660.6 [ (M+Na)+] .
Step 3.e: Preparation of (benzyl 2,4-di-O-acetyl-3-O-
benzyl-α-L-idopyranosyluronate) - (1-4) - (6-O-acetyl-2-
azido-3-O-benzyl-2-deoxy-α-D-glucopyranosyl) - (1-4) -
(benzyl 2-O-acetyl-3-O-benzyl-α-L-idopyranosyluronate) -
(1-4) - (6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-α-D-
glucopyranosyl)-(1-4)-((3S,4R,5R)-dibenzyl 5-(benzyl-
oxy)-4-oxypiperidine-1,3-carboxylate) (No. 16)
Compounds 15 (217 mg, 0.132 mmol) obtained in
step 3.d, and 6 (126 mg, 0.264 mmol) obtained in step
l.e, are treated according to METHOD 2 to give, after
purification, compound 16 (168 mg, 66%).
Mass spectrum (ESI) m/z 1976.0 [(M+Na)+].
Step 3.f: Preparation of (benzyl 2,4-di-O-acetyl-3-O-
benzyl-α-L-idopyranosyluronate)-(1-4)-(6-O-acetyl-2-
acetamido-3-O-benzyl-2-deoxy-α-D-glucopyranosyl) - (1-4) -
(benzyl 2-O-acetyl-3-O-benzyl-α-L-idopyranosyluronate) -
(1-4) -(6-O-acetyl-2-acetamido-3-O-benzyl-2-deoxy-α-D-
glucopyranosyl)-(1-4)-((3S,4R,5R)-dibenzyl 5-(benzyl-

oxy)-4-oxypiperidine-1,3-dicarboxylate) (No. 17)
Compound 16 (138.5 mg, 70.9 µmol) obtained in
step 3.e is dissolved in pyridine. (1.16 mL) and then
thioacetic acid (1.14 mL, 225 molar equivalents) is
added at 0°C. The reaction medium is stirred for 14 h
at room temperature and is then concentrated and
purified on silica gel (3:2 cyclohexane-ethyl acetate)
to give compound 17 (118 mg, 84%).
Mass spectrum (ESI) m/z 2007.7 [ (M+Na)+] .
Step 3.g: Preparation of (3-O-benzyl-α-L-
idopyranosyluronic acid)-(1-4)-(2-acetamido-3-O-benzyl-
2-deoxy-α-D-glucopyranosyl) - (1-4) - (3-O-benzyl-α-L-
idopyranosyluronic acid)-(1-4)-(2-acetamido-3-O-benzyl-
2-deoxy-α-D-glucopyranosyl) - (1-4) - ( (3S,4R, 5R) benzyl
5-(benzyloxy)-4-oxypiperidine-3-carboxylic acid-1-
carboxylate) (No. 18)
Compound 17 (101 mg, 50.9 µmol.) obtained in
step 3.f is treated according to METHOD 3 and then the
reaction medium is acidified with 6N hydrochloric acid
(pH 1) and extracted with dichloromethane . The organic
phase is washed with 5% sodium sulphite (Na2SO3) and
then with water. After drying, filtration and
concentration, the residue is used in the crude state
in the next step.
Mass spectrum (ESI) m/z 1505.6 [(M+H)+] .
Step 3.h: Preparation of (3-O-benzyl-2,4-di-O-sodium
sulphonato-α-L-idopyranosyluronate of sodium)-(1-4)-(2-

acetamido-3-O-benzyl-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranosyl)-(1-4)-(3-O-benzyl-2-O-sodium
sulphonato-α-L-idopyranosyluronate of sodium)-(1-4)-(2-
acetamido-3-O-benzyl-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranosyl)-(1-4)-(5-(benzyloxy)-4-oxypiperidine-1-
carboxylate of benzyl-3-carboxylate of sodium
(3S,4R,5R)) (No. 19)
The crude compound 18 obtained in step 3.g is
treated according to METHOD 4 to give compound 19
(50 mg, 54% (2 steps)).
Mass spectrum (ESI) m/z 2014 [ (M-3Na+3H) -] .
PREPARATION 4:
Synthesis of (3~O-benzyl-2,4-di-O-sodium sulphonato-α-
L-idopyranosyluronate of sodium)-(1-4)-(3-O-benzyl-2-
deoxy-2-N-sodium sulphonato-6-O-sodium sulphonato-α-D-
glucopyranosyl)-(1-4)-(3-O-benzyl-2-O-sodium
sulphonato-α-L-idopyranosyluronate of sodium)-(1-4)-(3-
O-benzyl-2-deoxy--2-N-sodium sulphonato-6-O-sodium
sulphonato-α-D-glucopyranosyl)-(1-4)-(5-(benzyloxy)-4-
oxypiperidine-1-carboxylate of benzyl-3-carboxylate of
sodium (3S,4R,5R)) (No- 26)


Step 4.a: Preparation of (3-O-benzyl-α-L-
idopyranosyluronic acid) - (1-4) - (2-azido-3-0,-benzyl-2-
deoxy-α-D-glucopyranosyl) - (1-4) - (3-O-benzyl-α-L-
idopyranosyluronic acid)- (1-4)- (2-azido-3-O-benzyl-2-
deoxy-α-D-glucopyranosyl)-(1-4)-(5-(benzyloxy)-4-
oxypiperidine-1-carboxylate of benzyl-3-carboxylate of
sodium (3S, 4R,5R)) (No. 24)
Compound 16 (175 mg, 89.6 µmol) obtained in

step 3.e is treated according to METHOD 3 and then the.
reaction medium is acidified with 6N hydrochloric acid
®
(pH 2). The mixture is then purified on a Sephadex
LH-20 column (1:1 dichloromethane-ethanol) to give
compound 24 (100 mg, 76%).
Mass spectrum (ESI) m/z 1474.1 [ (M+H)+] .
Step 4.b: Preparation of (3-O-benzyl-α-L-
idopyranosyluronic acid)-(1-4)-(2-amino-3-O-benzyl-2-
deoxy-α-D-glucopyranosyl)-(1-4)-(3-O-benzyl-α-L-
idopyranosyluronic acid)-(1-4)-(2-amino-3-O-benzyl-2-
deoxy-α-D-glucopyranosyl)-(1-4)-(5-(benzyloxy)-4-
oxypiperidine-1-carboxylate of benzyl-3-carboxylic acid
(3S,4R,5R)) (No. 25)
A 10% Pd/C/ethylenediamine complex prepared
according to the method described in H. Sajiki et-al.,
J. Org. Chem. (1998), Vol. 63, p. 799O-7992) (107 mg)
is added to a solution of compound 24 (3 5 mg) obtained
in step 4.a, in a 1:1 methanol-tetrahydrofuran mixture
(1 ml) . The medium is then placed under a H2 pressure
(3 bar) at room temperature for 16 h. After filtration
and concentration, the crude reaction product is
reacted again under the same conditions and is then
purified on silica gel (ethyl acetate-pyridine-acetic
acid-water, 6:2:0.6:1) to give compound 25 (12 mg,
44%) .
Mass spectrum (ESI) m/z 1421.4 [(M+H)+]
Step 4.C: Preparation of (3-O-benzyl-2,4-di-O-sodium

sulphonato-α-L-idopyranosyluronate of sodium)-(1-4)-(3-
O-benzyl-2-deoxy-2-N-sodium sulphonato-6-OTsodium Sulphonato-α-D-glucopyranosyl) - (1-4) -(3-O-benzyl-2-O-
sodium sulphonato-α-L-idopyranosyluronate of sodium)-
(1-4)-(3-O-benzyl-2-deoxy-2-N-sodium,sulphonato-6-O-
sodium sulphonato-α-D-glucopyranosyl)-(1-4) -(5-
(benzyloxy)-4-oxypiperidine-1-carboxylate of benzyl-3-
carboxylate of sodium (3S,4R,5R)) (No. 26)
Compound 25 (8.5 mg, 5.98 µmol) obtained in
step 4.b is treated according to method 4 to give
compound 2 6 (7 mg, 56%).
Mass spectrum (ESI) m/z 2133.8 [ (M-H)-] .
PREPARATION 5:
Synthesis of (benzyl 2-O-acetyl-4-O-levulinoyl-3-O-
methyl-α-L-idopyranosyluronate)-(1-4)-(6-O-acetyl-2-O-
(3-bromo-4-methoxy)benzyl-3-O-methyl-α, β-D-gluco-
pyranose trichloroacetimidate) (No. 40)


Step 5.a: Preparation of l,,6-anhydro-4-O-
(tetrahydropyran-2-yl)-2-O-(4-methoxy)benzyl-β-D-
glucopyranose (No. 29)
Sodium (1.37 g, 0.33 molar equivalent) is
added, at 40°C, to a solution of compound 28 (41 g,
18 0 mmol) (prepared according to H. Paulsen and W.
Stenzel, Chem. Ber. (1978) 111, 2348-57) in para-
methoxybenzyl alcohol (25 ml, 1.1 molar equivalent).
The reaction mixture is then heated at 110°C for 20 min
and then methanol (2 0 ml) is added at 0°C and the .
stirring is maintained for 16 h at room temperature.
After concentration, purification on silica gel (3:7
ethyl acetate-diisopropyl ether) gives 29 (20.1 g,
31%) .

Mass spectrum (ESI) m/z 384.2 [(M+Na)+].
Step 5.b: Preparation of 1,6-anhydro-4-O-
(tetrahydropyran-2-yl)-2-O-(4-methoxy)benzyl-3-O-
methyl-β-D-glucopyranose (No. 30)
Methyl iodide (2.67 ml) and sodium hydride
(2.68 g) are successively added, at 0°C and under an
argon atmosphere, to a solution of compound 29 (13.1 g,
35.8 mmol) obtained in step 5.a, in dimethylformamide
(10 7 ml). After returning to room temperature, the
reaction mixture is stirred for 16 h and then methanol
is added at 0°C, the medium is extracted with ethyl
acetate, dried (Na2SO4) , filtered and concentrated to
give 30 which is directly used in the next step.
Mass spectrum (ESI) m/z 403.3 [ (M+Na)+] .
Step 5.c: Preparation of 1,6-anhydro-2-O-(4-
methoxy)benzyl-3-O-methyl-β-D-glucopyranose (No. 31)
A 0.25M methanolic solution of camphor
sulphonic acid (CSA) (1 molar equivalent) is added to a
solution of the crude compound 30 obtained in step 5.b,
in methanol (143 ml). After 3 0 min of magnetic
stirring, the medium is diluted with dichloromethane
and then washed with water, with a 2% aqueous sodium
hydrogen carbonate solution, with water, dried (Na2SO4) ,
filtered and concentrated. Purification on silica gel
(3:7 ethyl acetate-cyclohexane) gives compound 31
(9.0 g, 85%).
Mass spectrum (ESI) m/z 319.1 [(M+Na)+].

Step 5.d: Preparation of ethyl 2-Q-acetyl-4,6-O-
isopropylidene-3-O-methyl-1-thio-α, β-L -idopyranoside
(No. 33)
Triethylamine (2.2 ml), DMAP (173 mg) and
acetic anhydride (1.34 ml) are successively added, at
0°C, under argon, to a solution of the crude compound
32 (1.99 g, 7.1 mmol) (prepared according to
P. Duchaussoy et al., Carbohydr. Res. (1999), 317,
63-84) in dichloromethane (37 ml). The temperature is
kept at 0°C for 10 min and then the reaction medium is
placed at room temperature for 16 h. The reaction
mixture is then concentrated under vacuum and the
residue purified on silica (1:1 Et2O-cyclohexane) to
give compound 33 (2.1 g, 92%).
Mass spectrum (ESI) m/z 343.3 [(M+Na)+] .
Step 5.e: Preparation of (2-O-acetyl-4,6-O-isoprop-
yl idene-3-O-methyl-α-L-idopyrandsyl) -(1-4) - (1,6-
anhydro-2-O-(4-methoxy)benzyl-3-O-methyl-β-D-gluco-
pyranose) (No. 34a?) and (2-O-acetyl-4,6-O-isoprop-
ylidene-3-O-methyl-β-L-idopyranosyl) - (1-4) - (1,6-
anhydro-2-O- (4-methoxy) benzyl-3-O-methyl-β-D-gluco-
pyranose) (No. 34/3)
A solution of N-iodosuccinimide (1.38 g) and
trifluoromethanesulphonic acid (63.5 µl) in a 1:1
dichloromethane-dioxane mixture (16.5 ml) is added, at
-2 0°C, to a mixture, under argon, of compound 33
(1.86 g, 5.79 mmol) obtained in step 5.d, and of

compound 31 (1.46 g, 4.94 mmol) obtained in step 5.c,
in the presence of 4 A molecular sieves (2.89 g) in
toluene (50 ml). After stirring for 45 min, solid
sodium hydrogen carbonate is added to the reaction
medium, and after filtration, the mixture is diluted
with dichloromethane, washed with 10% aqueous sodium
thiosulphate (Na2S2O3) solution and a saturated aqueous
sodium chloride solution. After drying and
concentrating, the residue is directly used in the next
step.
Mass spectrum (ESI) m/z 577.4 [ (M+Na)+] .
Step 5.f: Preparation of (2-O-acetyl-3-O-methyl-α-L-
idopyranosyl)-(1-4)-(1,6-anhydro-2-O-(4-methoxy)benzyl-
3-O-methyl-β-D-glucopyranose) (No. 35a) and (2-O-
acetyl-3-O-methyl-β-L-idopyranosyl) - (1-4) - (1, 6-anhydro-
2-O- (4-methoxy) benzyl-3-O-methyl-β-D-glucopyranose)
(No. 35/3)
Acetic acid at 70% (55 ml) is added to a
solution of the mixed compound 34a and 34/? obtained in
step 5.e, in 1,2-dichloroethane (12 ml). The mixture is
heated at 60°C for 50 min and then concentrated under
vacuum and the residue obtained is purified on silica
gel (3:2 toluene-acetone) to give compound 35a (1.56 g,
61%, two steps) , and compound 35/? (0.36 g, 14%, two
steps).
Mass spectrum (ESI) m/z 537.5 [(M+Na)+] .
Step 5.g: Preparation of (benzyl 2-O-acetyl-3-O-methyl-

α-L-idopyranosyluronate)- (1-4)-(1,6-anhydro-2-O-(3-
bromo-4-methoxy)benzyl-3 -O-methyl-fi-D-glucopyranose)
(No. 36)
Compound 35a (0.975 g, 1.89 mmol) obtained in
step 5f is treated according to METHOD 1 to give, after
purification on silica gel (1:1 ethyl acetate-
cyclohexane), compound 36 (1.09 g, 83%).
1H NMR (CDC13) δ7.54-6.86 (m, 8H, Ar).
Step 5.h: Preparation of (benzyl 2-O-acetyl-4-O-
levulinoyl-3-O-methyl-α-L-idopyranosyluronate) - (1-4) -
(1, 6-anhydro-2-O- (3-bromo-4-methoxy) benzyl-3-O-me,thyl-
β-D-glucopyranose) (No. 37)
DMAP (43 mg, 0.2 molar equivalent), 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
(EDC1) (0.675 g, 2 molar equivalents) and levulinic
acid (0.361 ml, 2 molar equivalents) are successively
added to a solution, under argon, of compound 36
(1.09 g, 1.56 mmol) obtained in step 5.g, in dioxane
(35 ml), and after stirring for 16 h, the reaction
medium is successively washed with a 10% potassium
hydrogen sulphate (KHSO4) solution, water, 2% sodium
hydrogen carbonate and then the solution is dried
(Na2SO4) , filtered and concentrated to give a residue
which is purified on silica gel (ethyl acetate-
dichloromethane 2:3) giving compound 37 (1.07 g, 86%).
Mass spectrum (ESI) m/z 819.4 [ (M+Na)+] .
Step 5.i: Preparation of (benzyl 2-O-acetyl-4-O-

levulinoyl-3-O-methyl-α-L-idopyranosyluronate) - (1-4) -
(1,6-di-O-acetyl-2-O-(3-bromo-4-methoxy)benzyl-3-O-
methyl-α, β-D-glucopyranose) (No. 38)
Compound 37 (1.07 g, 1.34 mmol) obtained in
step 5.h is treated as for the synthesis of compound 9
(step 2.b) to give, after purification on silica gel
(ethyl acetate-dichloromethane 2:3), compound 38
(1.04 g, 87%).
Mass spectrum (ESI) m/z 921.4 [ (M+Na)+] .
Step 5.j: Preparation of (benzyl 2-O-acetyl-4-O-
levulinoyl-3-O-methyl-α-L-idopyranosyluronate) - (1-4) -
(6-O-acetyl-2-O-(3-bromo-4-methoxy)benzyl-3-O-methyl-
a, β-D-glucopyranose) (No. 39)
Compound 38 (1.04 g, 1.16 mmol) obtained in
step 5.i is treated as for the synthesis of compound 10
(step 2.c) to give, after purification on silica gel
(ethyl acetate-dichloromethane 1:1), compound 39
(740 mg, 74%).
Mass spectrum (ESI) m/z 877.3 [(M+Na)+] .
Step 5.k: Preparation of (benzyl 2-O-acetyl-4-O-
levulindyl-3-O-methyl-α-L-idopyranosyluronate) - (1-4) -
(6-O-acetyl-2-O-(3-bromo-4-methoxy)benzyl-3-O-methyl-
a, β-D-glucopyranose trichloroacetimidate) (No. 40)
Compound 39 (74 0 mg, 8 60 µmol) obtained in
step 5.j is treated as for the synthesis of compound 11
to give, after purification on silica gel (ethyl
acetate-dichloromethane 3:7), compound 40 (714 mg,

8.3%) .
1H NMR (CDCl3) δ6.39 (d, H-1α GlC1) , 5.76 (d, H-lβ GlC1) . PREPARATION 6:
Synthesis of (3-O-methyl-2,4-di-O-sodium sulphonato-α-
L-idopyranosyluronate of sodium)-(1-4)-(3-O-methyl-2,6-
di-O-sodium sulphonato-α-D-glucopyranosyl)-(1-4)-(3-O-
methyl-2-O-sodium sulphonato-α-L-idopyranosyluronate of
sodium)-(1-4)-(3-O-methyl-2,6-di-O-sodium sulphonato-α-
D-glucopyranosyl) -(1-4)-(5-(benzyloxy)-4-oxypiperidine-
1-carboxylate of benzyl-3-carboxylate of sodium
(3S,4R,5R)) (compound No. 46)


Step 6.a: Preparation of (benzyl 2-O-acetyl-4-O-
levulinoyl-3-O-methyl-α-L-idopyranosyluronate) - (1-4) -
(6-O-acetyl-2-O- (3-bromo-4-methoxy) benzyl-3-O-methyl-α-
D-glucopyranosyl) - (.1-4) - ( (3S,4R,5R) -dibenzyl 5- (benzyl-
oxy)-4-oxypiperidine-1,3-dicarboxylate) (No. 41)
A mixture of compound 40 (94 mg, 0.094 mmol,
1.0 molar equivalent) obtained in step 5.k, and of
compound 6 (111 mg, 0.234 mmol, 2.5 molar equivalents)
obtained in step l.e, is treated according to METHOD 2
to give, after purification on Sephadex LH-20 column,
and then on silica gel (dichloromethane-ethyl acetate
9:1), compound 41 (62 mg, 50%)...
Mass spectrum (ESI) m/z 1336.5 [ (M+Na)+] .
Step 6.b: Preparation of (benzyl 2-O-acetyl-3-O-methyl-
α-L-idopyranosyluronate)-(1-4)-(6-O-acetyl-2-O-(3-
bromo-4-methoxy)benzyl-3-O-methyl-α-D-glucopyranosyl)-
(1-4)-((3S,4R,5R)-dibenzyl 5-(benzyloxy)-4-
oxypiperidine-1,3-dicarboxylate) (No. 42)
Hydrazine acetate (21 mg, 10 molar
equivalents) is added to a solution of compound 41
(60 mg, 45.7 µmol) obtained in step 6.a, in a 1:2

toluene/ethanol mixture (9 ml). After 3 h of magnetic
stirring, the mixture is concentrated under vacuum and
the residue is diluted with dichloromethane, washed
with a 2% sodium hydrogen carbonate solution, with
water, and the organic phase is dried (Na2SO4) , filtered
and then concentrated. The residue is purified on

silica gel (ethyl acetate-cyclohexane 3:2) to give
compound 42 (48 mg, 88%).
Mass spectrum (ESI) m/z 1238.5 [(M+Na)+] .
Step 6.c: Preparation of (benzyl 2-O-acetyl-4-O-
levulinoyl-3-O-methyl-α-L-idopyranosyluronate)-(1-4) -
(6-O-acetyl-2-O-(3-bromo-4-methoxy)benzyl-3-O-methyl-α-
D-glucopyranosyl) - (1-4) - (benzyl 2-O-acetyl-3-O-methyl-
α-L-idopyranosyluronate) - (1-4) - (6-O-acetyl-2-O- (3-..
bromo-4-methoxy) benzyl-3-O-methyl-α-D-glucopyranosyl) -
(1-4)-((3S,4R,5R)-dibenzyl 5-(benzyloxy)-4-
oxypiperidine-1,3-dicarboxylate) (No. 43)
A mixture of compound 40 (197.7 mg,
197.7 µmol, 1.14 molar equivalents) obtained in
step 5.k, and of compound 42 (210.5 mg, 173.2 µmol,
1.0 molequiv.) obtained in step 6.b, is treated
according to method 2 to give, after purification on a
Sephadex LH-20 column, and then on silica gel (toluene-
acetone 1:1), compound 43 (176 mg, 50%).
Mass spectrum (ESI) m/z 2075.0 [ (M+Na)+] .
Step 6.d: Preparation of (benzyl 2-O-acetyl-4-Q-
levulinoyl-3-O-methyl-α-L-idopyranosyluronate)-(1-4)-
(6-O-acetyl-3-O-methyl-α-D-glucopyranosyl) - (1-4) -
(benzyl 2-O-acetyl-3-O-methyl-α-L-idopyranosyluronate) -
(1-4) - (6-O-acetyl-3-O-methyl-α-D-glucopyranosyl) - (1-4) -
((3S,4R,5R)-dibenzyl 5-(benzyloxy)-4-oxypiperidine-1,3-
dicarboxylate) (No. 44)
Zirconium tetrachloride (ZrCl4) (39 mg,

5 molar equivalents) is added, at 0°C and under an
argon atmosphere, to a solution of compound 43 (69 mg,
33.6 µmol) obtained in step 6.c, in acetonitrile
(3 ml). After stirring for 45 min at room temperature,
the mixture is concentrated under vacuum and the
residue is diluted with ethyl acetate, washed with
water, and after drying, filtration and concentration,
the residue is purified on silica gel (3:7 acetone-
toluene) to give compound 44 (52 mg, 89%).
Mass spectrum (ESI) m/z 1676.7 [(M+Na)+].
Step 6.e: Preparation of (3-O-methyl-α-L-
idopyranosyluronic acid)-(1-4)-(3-O-methyl-α-D-
glucopyranosyl)-(1-4)-(3-O-methyl-α-L-
idopyranosyluronic acid)-(1-4)-(3-O-methyl-α-D-
glucopyranosyl)-(1-4)-(5-(benzyloxy)-4-oxypiperidine-1-
carboxylate of benzyl-3-carboxylic acid (3S,4R,5R))
(No. 45)
Compound 44 (18 mg, 11 µmol) obtained in
step 6.d is treated according to METHOD 3 to give,
after purification, compound 45 (5.8 mg, 47%) which may
be partially esterified on the carboxylic acid groups.
Mass spectrum (ESI) m/z 1118.4 [(M+H)+] .
Step 6.f: Preparation of (3-O-methyl-2,4-di-O-sodium
sulphonato-α-L-idopyranosyluronate of sodium)-(1-4) -(3 -
O-methyl-2,6-di-O-sodium sulphonato-α-D-gluco-
pyranosyl)-(1-4)-(3-O-methyl-2-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(3-O-methyl-2,6-

di-O-sodium sulphonato-α-D-glucopyranosyl)-(1-4)-(5-
(benzyloxy)-4-oxypiperidine-1-carboxylate of benzyl-3-
carboxylate of sodium (3S,4R,5R)) (No. 46)
Compound 45 (7 mg, 6.26 µmol) obtained in
step 6.e is treated according to.METHOD 4 to give,
after purification, compound 46 (10 mg, 77%) which may
be partially esterified on the carboxylic acid groups.
Mass spectrum (ESI) m/z 189.7.0 [ (M+Na-H)+] .
PREPARATION 7:
Synthesis of (benzyl 2-O-acetyl-4-O-levulinoyl-3-O-
benzyl-ot-L-idopyranosyluronate) - (1-4) - (6-O-acetyl-2-O-
(4-methoxy) benzyl-3 -O-benzyl-a, β-D-glucopyranose
trichloroacetimidate) (No. 57)


(tetrahydropyran-2-yl)-2-O-(4-methoxy)benzyl-3-O-
benzyl-β-D-glucopyranose (No. 48)
Benzyl bromide (5.1 ml) and then sodium
hydride (4.6 g) are added, at 0°C, to a solution of 29
(19.96 g, 54.5 mmol) obtained in step 5.a, in DMF
(300 ml). At the end of the addition, the mixture is
placed at room temperature for 16 h and then methanol
(18 ml) is added at 0°C, and after stirring for 1 h at
room temperature, the medium is diluted with ethyl
acetate (600 ml), washed with water (300 ml), dried
(Na2SO4) , filtered and concentrated. The residue is
purified by flash chromatography (5:95 ethyl acetate-
diisopropyl ether) to give 48 (20.6 g, 83%).
Mass spectrum (ESI) m/z 479.3 t(M+Na)+].
Step 7.b: Preparation of 1,6-anhydro-2-O-(4-methoxy)-
benzyl-3-O-benzyl-β-D-glucopyranose (No. 49)
Compound 48 (20.6 g, 45.2 mmol) obtained in
step 7.a is treated as for the synthesis of compound 31
(step 5.c) to give,, after purification on silica (3:7
ethyl acetate-cyclohexane), 49 (14.4 g, 86%).
Mass spectrum (ESI) m/z 395.4 [ (M+Na)+] .
Step 7.c: Preparation of (2-O-acetyl-4,6-O-
isopropylidene-3-O-benzyl-ce, β-L-idopyranosyl) - (1-4) -
(1, 6-anhydro-2-O- (4-methoxy) benzyl -3 -O-benzyl-β-D-
glucopyranose) (No. 51)
Compound 50 (prepared according to the method
described by C. Tabeur et al. for the 2-O-benzoylated

derivative, Carbohydr. Res. (1996), 281, 253-276)
(16.91 g, 42.6 mmol) and compound 49 (14.44 g,
38.8 mmol) obtained in step 7.b are reacted as for the
synthesis of 34 (step 5.e) to give, after purification
on silica (15:85 ethyl acetate-diisopropyl ether),
compound 51 (17.05 g, 62% (56% alpha-L)).
Mass spectrum (ESI) m/z 729.3 [ (M+Na)+] .
Step 7.d: Preparation of (2-O-acetyl-3-O-benzyl-α-L-
idopyranosyl)-(1-4)-(1,6-anhydro-2-O-(4-methoxy)benzyl- 3-O-benzyl-β-D-glucopyranose) (No. 52)
Compound 51 (12.38, 17.51 mmol) obtained in
step 5.c is treated as for the synthesis of 35
(step 5.f) to give, after purification on silica (4:1
toluene-acetone), 52 (10.85 g, 93%).
Mass spectrum (ESI) m/z 689.3 [(M+Na)+].
Step 7.e: Preparation of (benzyl 2-O-acetyl-3-O-benzyl-
α-L-idopyranosyluronate)-(1-4)-(1,6-anhydro-2-O-(4-
methoxy)benzyl-3-O-benzyl-β-D-glucopyranose) (No. 53)
Compound 52 (10.85 g, 16.2 7 mmol) obtained in
step 7.d is treated according to METHOD 1 to give,
after purification on silica (3:7 acetone-cyclohexane),
53 (8.44 g, 61%).
Mass spectrum (ESI) m/z 793.3 [(M+Na)+.
Step 7.f: Preparation of (benzyl 2-O-acetyl-3-O-benzyl-
4-O-levulinoyl-α-L-idopyranosyluronate) - (1-4) -.(1,6-
anhydro-2-O-(4-methoxy)benzyl-3-O-benzyl-β-D-
glucopyranose) (No. 54)

Compound 53 (3.2 g, 3.77 mmol) obtained in
step 7.e is treated as for the synthesis of 37
(step 5.h) to give 54 (3.17 g, 89%) after purification
on silica (acetone-toluene 1:4).
Mass spectrum (ESI) m/z 891.3 [ (M+Na)+] .
Step 7.g: Preparation of (benzyl 2-O-acetyl-4-O-
levulinoyl-3-O-benzyl-α-L-idopyranosyluronate)-(1-4)-
(1, 6-di-O-acetyl-2-O- (4-methoxy) benzyl-3-O-benzyl-α, β-
D-glucopyranose) (No. 55)
Compound 54 (1.45 g, 1.53 mmol) obtained in
step 7.f is treated as for the synthesis of 9 but at
0°C for 1 h to give, after purification on silica
(toluene-acetone 85:15), 55 (1.25 g, 78%).
Mass spectrum (ESI) m/z 993.3 [(M+Na)+].
Step 7.h: Synthesis of (benzyl 2-O-acetyl-4-O-
levulinoyl-3-O-benzyl-g;-L-idopyranosyluronate) - (1-4) -
(6-O-acetyl-2-O-(4-methoxy)benzyl-3-O-benzyl-g,β-D-
glucopyranose) (No. 56)
Compound 55 obtained in step 7.g is treated
as for the synthesis of 10 (step 2.c) to give, after
purification on silica (diethyl ether-dichloromethane
25:75), 56 (429 mg, 56%).
Mass spectrum (ESI) m/z 951.3 [ (M+Na)+] .
Step 7.i: Preparation of (benzyl 2-O-acetyl-4-O-
levulinoyl-3-O-benzyl-α-L-idopyranosyluronate) - (1-4) -
(6-O-acetyl-2-O-(4-methoxy)benzyl-3-O-benzyl-a,β-D-
glucopyranose trichloroacetimidate) (No. 57)

Compound 56 (429 mg, 426 µmol) obtained in
step 7.h is treated as for the synthesis of 11
(step 2.d) to give, after purification on silica (ethyl
acetate-cyclohexane 1:1), the derivative 57 (396 mg,
80%) .
1H NMR (CDC13) 8 6.45 (d, H-la) , 5.89 (d, H-1β.
PREPARATION 8:
Preparation of (benzyl 2-O-acetyl-4-O-allyl-3-O-benzyl-
α-L-idopyranosyluronate)-(1-4)-(6-O-acetyl-2-O-(4-
methoxy)benzyl-3-O-benzyl-α,β-D-glucopyranose
trichloroacetimidate) (No. 62)

Step 8.a: Preparation of (benzyl 2-O-acetyl-4-O-
allyloxycarbonyl-3-O-benzyl-α-L-idopyranosyluronate)-
(1-4) - (1, 6-anhydro-2-O- (4-methoxy) benzyl-3-O-benzyl-β-
D-glucopyranose)(No. 58)
Pyridine (759 µ1, 9.41 mmol) , DMAP (115 mg,
0.94 mmol) and allyl chloroformate (995 µl, 9.41 mmol)
in. solution in THF (2.35 ml) are added, at 0°C and
under an argon atmosphere, to a solution of 53
(0.800 g, 0.9414 mmol) obtained in step 7.e, in THF

(9.4 ml). The stirring is maintained overnight and then
the reaction mixture is diluted with ethyl acetate,
washed with 10% KHSO4, with 2% sodium hydrogen
carbonate, with water, dried (Na2SO4) , filtered and
concentrated. The residue is purified by flash
chromatography (1:9 acetone-toluene), to give
58 (0.809 g, 92%).
Mass spectrum (ESI) m/z 877.3 [ (M+Na)+] .
Step 8.b: Preparation of (benzyl 2-O-acetyl-4-O-allyl-
3-O-benzyl-α-L-idopyranosyluronate) - (1-4) - (1, 6-anhydro-
2-O- (4-methoxy) benzyl-S-O-benzyl-β-D-glucopyranose)
(No. 59)
Palladium diacetate (3.9 mg, 0.017 mmol) and
triphenylphosphine (22.6 mg, 0.086 mmol) are
successively added, under an argon atmosphere, to a
solution, in THF (6 ml), of compound 58 (0.805 g,
0.8 62 mmol) obtained in step 8.a. The temperature of
the mixture is brought to 90°C for 15 min and then the
medium is concentrated under vacuum and purified on
silica (15:85 acetone-toluene) to give 59 (0.587 g,
66%) .
Mass spectrum (ESI) m/z 833.4 [(M+Na)+].
Step 8.c: Preparation of (benzyl 2-O-acetyl-4-O-allyl-
3-O-benzyl-α-L-idopyranosyluronate)-(1-4)-(1,6-di-O-
acetyl-2-O- (4-methoxy) benzyl-3-O-benzyl-a?, β-D-
glucopyranose) (No. 60)
Compound 59 (0.587 g, 0.660 mmol) obtained in

step 8.b is treated as for the synthesis of compound 55
(step 7.g) to give, after purification on silica (9:1
acetone-toluene), 60 (0.37 g, 57%).
Mass spectrum (ESI) m/z 936.4 [ (M+Na)+] .
Step 8.d: Preparation of (benzyl 2-O-acetyl-4-O-allyl-
3-O-benzyl-α-L-idopyranosyluronate) - (1-4) - (6-O-acetyl-
2-O- (4-methoxy) benzyl-3-O-benzyl-a, β-D-glucopyranose)
(No. 61)
Compound 60 obtained in step 8.c is treated
as for the synthesis of 10 (step 2.c) to give, after
purification on silica gel (cyclohexane-ethyl acetate
2:3), 61 (240 mg, 70%).
Mass spectrum (ESI) m/z 893.4 [ (M+Na)+] .
Step 8.e: Preparation of (benzyl 2-O-acetyl-4-allyl-3-
O-benzyl-α-L-idopyranosyluronate) - (1-4) - (6-O-acetyl-2-
O-(4-methoxy)benzyl-3-O-benzyl-a,β-D-glucopyranose
trichloroacetimidate) (No. 62)
Compound 61 (234 mg, 246 µmol) obtained in
step 8.d is treated as for the synthesis of 11
(step 2.d) to give, after purification on silica (86:14
acetone-toluene), the derivative 62 (249 mg, 93%).
1H NMR (CDC13) 8 6.40 (d, H-lα) , 5.81 (d, H-1β) .
The syntheses of PREPARATIONS 9 and 10 may be
schematically represented as follows:


PREPARATION 9:
Synthesis of (3-O-benzyl-2,4-di-O-sodium sulphonato-α-
L-idopyranosyluronate of sodium)-(1-4)-(3-O-benzyl-2,6-
di-O-sodium sulphonato-α-D-glucopyranosyl) - (1-4) - (3-O-
benzyl-2-O-sodium sulphonato-α-L-idopyranosyluronate of
sodium) - (1-4) - (3-O-benzyl-2, 6-di-O-sodium sulphonato-α-
D-glucopyranosyl)-(1-4)-(5-(benzyloxy)-4-oxypiperidine-
1-carboxylate of benzyl-3-carboxylate of sodium
(3S,4R,5R)) (No. 68)
Step 9.a: Preparation of (benzyl 2-O-acetyl-4-O-

levulinoyl-3-O-benzyl-α-L-idopyranosyluronate)-(1-4) -
(6-O-acetyl-2-O- (4-methoxy) benzyl-3-O-benzyl-α-D-
glucopyranosyl)-(1-4)-((3S,4R,5R)-dibenzyl 5-
(benzyloxy)-4-oxypiperidine-1,3-dicarboxylate) (No. 63)
Compound 57 (396 mg, 0.34 mmol) obtained in
step 7.i, and compound 6 (405 mg, 0.85 mmol) obtained
in step l.e, are treated according to METHOD 2 to give,
after purification, 63 (369 mg, 73%).
1H NMR (CDC13) δ5.29 (d, H-l GlcII) , 5.13 (d, H-1
IdoUAIII) ...
Step 9.b: Preparation of (benzyl 2-O-acetyl-3-O-benzyl-
α-L-idopyranosyluronate)-(1-4)-(6-O-acetyl-2-O-(4-
methoxy) benzyl-3-O-benzyl-α-D-glucopyranosyl) - (1-4) -
( (3S,.4R, 5R) -dibenzyl 5- (benzyloxy) -4-oxypiperidine-1, 3-
dicarboxylate) (No. 64)
Compound 63 (372 mg, 0.254 mmol) obtained in
step 9.a is treated as for the synthesis of 42
(step 6.b) to give, after purification on silica (ethyl
acetate-cyclohexane 2:3), compound 64 (301 mg, 87%).
XH NMR (CDCl3) δ 5.29 (d, H-l GlC11), 5.10 (d, H-l
IdoUA111), 3.97 (dd, H-4 IdoUA111).
Step 9.c: Preparation of (benzyl 2-O-acetyl-4-O-
levulinoyl-3-O-benzyl-α-L-idopyranosyluronate) - (1-4) -
(6-O-acetyl-2-O- (4-methoxy) benzyl-3-O-benzyl-α-D-
glucopyranosyl)-(1-4)-(benzyl 2-O-acetyl-3-O-benzyl-α-
L-idopyranosyluronate)-(1-4)-(6-O-acetyl-2-O-(4-
methoxy)benzyl-3-O-benzyl-α-D-gIucopyranosyl)-(1-4)-

((3S,4R,5R)-dibenzyl 5-(benzyloxy)-4-oxypiperidine-1,3-
dicarboxylate) (No. 65)
Compounds 57 (124 mg, 0.108 mmol) obtained in
step 7.i, and 64 (150 mg, 0.110 mmol) obtained in
step 9.b, are treated according to METHOD 2 to give,
after purification, 65 (90 mg, 35%).
1H NMR (CDC13) 8 5.28 (d, H-l GlcII) , 5.16 (d, H-1
IdoUAIII) , 5.13 (d, H-l IdoUAv) , 4.74 (d, H-l GlcIV) .
Step 9.d: Preparation of (benzyl 2-O-acetyl-4-O-
levulinoyl-3-O-benzyl-α-L-idopyranosyluronate) - (1-4) -
(S-O-acetyl-3-O-benzyl-α-D-glucopyranosyl)-(1-4)-
(benzyl 2-Q-acetyl-3-O-benzyl-α-L-idopyranosyluronate)-
(1-4)-(6-O-acetyl-3-O-benzyl-α-D-glucopyranosyl)-(1-4)-
((3S,4R,5R)-dibenzyl 5-(benzyloxy)-4-oxypiperidine-1,3-
dicarboxylate) (No. 66)
Compound 65 (45 mg, 0.19 mmol) obtained in
step 9.c is treated as for the synthesis of 44
(step 6.d) to give, after purification on silica (ethyl
acetate-cyclohexane 3:2), compound 66 (34 mg, 91%).
Mass spectrum (ESI) m/z 1982.0 [(M+Na)+] .
Step 9.e: Preparation of (3-O-benzyl-α-L-idopyranosyl-
uronic acid)-(1-4)-(3-O-benzyl-α-D-glucopyranosyl)-
(1-4) - (3-O-benzyl-α-L-idopyranosyluronic acid) - (1-4) -
(3-O-benzyl-α-D-glucopyranosyl)-(1-4)-(5-(benzyloxy)-4-
oxypiperidine-1-carboxylate of benzyl-3-carboxylic acid
(3S,4R,5R)) .(No. 67)
Compound 66 (25 mg, 12.8 µmol) obtained in

step 9.d is treated according to METHOD 3. The reaction
mixture is acidified with 6N hydrochloric acid (pH 2)
and then deposited on an LH-20 column (100 ml)
equilibrated in a 9:1 DMF/water mixture. The fractions
containing the product are then concentrated and
purified on silica (dichloromethane-methanol 7:3) to
give 67 (10.4 mg, 59%) which may be partially
esterified on the carboxylic acid groups.
Mass spectrum (ESI) m/z 1422.7 [ (M+H)+] .
Step 9.f: Preparation of (3-O-benzyl-2,4-di-O-sodium
sulphonato-α-L-idopyranosyluronate of sodium)-(1-4)-(3 -
O-benzyl-2,6-di-O-sodium sulphonato-α-D-gluco-
pyranosyl)-(1-4)-(3-O-benzyl-2-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(3-Q-benzyl-2,6-
di-O-sodium sulphonato-α-D-glucopyranosyl)-(1-4)-(5-
(benzyloxy)-4-oxypiperidine-1-carboxylate of benzyl-3-
carboxylate of sodium (3S,4R,5R))(No.68)
Compound 67 (10.4 mg, 9 µmol) obtained in
step 9.e is treated according to METHOD 4 to give 68
which is used directly in the next step.
PREPARATION 10:
Synthesis of (4-O-allyl-3-O-benzyl-2-O-sodium
sulphonato-or-L-idopyranosyluronate of sodium)-(1-4)-(3-
O-benzyl-2,6-di-O-sodium sulphonato-cx-D-
glucopyranosyl)-(1-4)-(3-O-benzyl-2-O-sodium
sulphonato-α-L-idopyranosyluronate of sodium)-(1-4)- (3 -
O-benzyl-2,6-di-O-sodium sulphonato-α-D-

glucopyranosyl)-(1-4)-(5-(benzyloxy)-4-oxypiperidine-1-
carboxylate of benzyl-3-carboxylate of sodium
(3S,4R,5R)). (No. 73)
Step 10.g: Preparation of (benzyl 2-O-acetyl-4-O-allyl-
3-O-benzyl-α-L-idopyranosyluronate)-(1-4)-(6-O-acetyl-
2-O-(4-methoxy)benzyl-3-O-benzyl-α-D-glucopyranosyl)-
(1-4)-(benzyl 2-O-acetyl-3-O-benzyl-α-L-idopyranosyl-
uronate)-(1-4)-(6-O-acetyl-2-O-(4-methoxy)benzyl-3-O-
benzyl-α-D-glucopyranosyl)-(1-4)-((35,4R,5R)-dibenzyl-
5-(benzyloxy)-4-oxypiperidine-1,3-dicarboxylate)
(No. 70)
Compound 62 (244 mg, 0.223 mmol) obtained in
step 8.e and compound 64 (138 mg, 0.101 mmol) obtained
in step 9.b are treated according to METHOD 2 to give,
after purification, 70 (100 mg, 43%).
1H NMR (CDC13) δ 5.28 (d, H-1 GlC11) , 5.25 (d, H-1
IdoUAv) , 5.16 (d, H-1 IdoUAIII) , 4.72 (d, H-1 GlcIV) .
Step 10.h: Preparation of (benzyl 2-O-acetyl-4-O-allyl-
3-O-benzyl-α-L-idopyranosyluronate) - (1-4) - (6-O-acetyl-
3-O-benzyl-α-D-glucopyranosyl)-(1-4)-(benzyl 2-O-
acetyl-S-O-benzyl-α-L-idopyranosyluronate)-(1-4) -(6-O-
acetyl-3-Q-benzyl-α-D-glucopyranosyl)-(1-4)-
((3S,4R,5R)-dibenzyl 5-(benzyloxy)-4-oxypiperidine-1,3-
dicarboxylate)(No. 71).
Compound 70 (92 mg, 40.0 µmol) obtained in
step 10.g is treated as for the synthesis of compound
44 (step 6.d) to give, after purification on silica

(acetone-toluene 17:83), compound 71 (44 mg, 59%).
Mass spectrum (ESI) m/z 1924.0 [ (M+Na)+] .
Step 10. i: Preparation of (4-O-allyl-3-O-benzyl-α-L-
idopyranosyluronic acid) - (1-4) - (3-O-benzyl-α-D-
glucopyranosyl) - (1-4) - (3-O-benzyl-α-L-idopyranosyl-
uronic acid)-(1-4)-(3-O-benzyl-α-D-glucopyranosyl)-(1-
4) -(5-(benzyloxy)-4-oxypiperidine-1-carboxylate of
benzyl-3-carboxylic acid (3S,4R,5R)) (No. 72)
Compound 71 (41 mg, .21.6 µmol) obtained in
step 10.h is treated according to method 3. The
reaction mixture is deposited on an LH-20 column
(210 ml) equilibrated in a 1:1 dichloromethane-ethanol
mixture. The fractions containing the product are then
concentrated and purified on silica to give 72
(30.3 mg, 96%) which may be partially esterified on the
carboxylic acid groups.
Mass spectrum (ESI) m/z 1462.4 [ (M+H)+] .
Step 10.j: Preparation of (4-O-allyl-3-O-benzyl-2-
sodium sulphonato-α-L-idopyranosyluronate of sodium)-
(1-4)-(3-O-benzyl-2,6-di-O-sodium sulphonato-α-D-gluco-
pyranosyl) - (1-4) - (3-O-benzyl-2-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(3-O-benzyl-2,6-
di-O-sodium sulphonato-α-D-glucopyranosyl)-(1-4)-(5-
(benzyloxy)-4-oxypiperidine-1-carboxylate of benzyl-3-
carboxylate of sodium (3S,4R,5R))(No. 73)
Compound 72 (10.0 mg, 6.44 µmol) obtained in
step 10.i is treated according to method 4 to give 73

which is used directly in the next step.
PREPARATION 11:
Synthesis of methyl (2-N-sodium sulphonato-2,4-dideoxy-
4-formyl-3, 6-di-O-sodium sulphonato-α-D-gluco-
pyranosyl)-(1-4)-(sodium 2-O-sodium sulphonato-α-L-
idopyranosyluronate)-(1-4)-(2-N-sodium sulphonato-2-
deoxy-6-O-sodium sulphonato-α-D-glucopyranoside)


(prop-1-en-1-yl) -β-D-mannopyranose. (No. 76)
Rhodium trichloride monohydrate (2 02 mg,
0.15 molar equivalent) is added, under argon, to a
solution of epoxide 75 (1.2 g, 7.14 mmol) (prepared
according to AG Kelly and JS Roberts, J. Chem. Soc,
Chem. Commun., (1980), Vol 288) in ethanol (56.5 ml).
After stirring for 1 h 25 min at 75°C, the reaction
medium is poured over 250 ml of ice-cold water, and
then after stirring for 5 min, the product is extracted
with diethyl ether, dried (Na2SO4) and concentrated. The
residue is then purified on silica (diisopropyl ether-
cyclohexane 45:55) and the fractions containing
compound 76 are partially concentrated (76 is
volatile.) .
1H NMR (CDC13) δ 3.42 (dd, H-2) , 3.00 (dd, H-3) , 2.64H
(dd, H-4).
Step 11.b: Preparation of 1,6-anhydro-2-azido-2,4-
dideoxy-4-(prop-1-en-1-yl)-β-D-glucopyranose (No. 77)
Compound 76 obtained in step 11.a is
dissolved in a dimethylformamide-water mixture (40 ml,
4:1) then sodium azide (7.0 g) is added, and the
mixture is heated under reflux for 10.5 h. The reaction
medium is then extracted with ethyl acetate, washed
with water and then with a saturated aqueous sodium
chloride solution, dried (Na2SO4) , concentrated and
purified on silica gel to give compound 77 (674 mg,
48%) .

1H NMR (CDCl3) δ 5.8-5.6 (m, 2H, CH=CH).
Step 11.c: Preparation of 1,3,6-tri-O-acetyl-2-azido-
2, 4-d'ideoxy-4- (prop-1-en-1-yl) -a, β-D-glucopyranose
(No. 78)
Compound 77 (3.5 g, 16.57 mmol) obtained in
step 11.b is treated as for the synthesis of compound . 9
(step 2.b) to give, after purification, compound 78
(5.88 g, 100%).
Mass spectrum (ESI) m/z 378 [ (M+Na)+] .
Step 11.d: Preparation of 3,6-di-O-acetyl-2-azido-2 , 4-
dideoxy-4-(prop-1-en-1-yl)-a,β-D-glucopyranose (No. 79)
Ethanolamine (4.0 ml, 4 molar equivalents) is
added, at 0°C, to a solution of compound 78 (5.88 g,
16.5 mmol) obtained in step 11.c, in tetrahydrofuran
(140 ml). After 16 h at +4°C, the medium is diluted
with ethyl acetate, acidified (1N HCl), washed with
water, dried (Na2SO4) and concentrated to give, after
purification, compound 79 (4.66 g, 90%).
Mass spectrum (ESI) m/z 336 [ (M+Na)+] .
Step 11.e: Preparation of 3,6-di-O-acetyl-2-azido-2,4-
dideoxy-4-(prop-1-en-1-yl)-a,β-D-glucopyranose
trichloroacetimidate (No. 80)
Potassium carbonate (K2CO3) (3.34 g, 1.6 molar
equivalents) and then CC13CN (7.6 ml, 5 molar
equivalents) are added, under argon, to a solution of
compound 79 (4.66 g, 14.9 mmol) obtained in step ll.d,
in dichloromethane (285 ml). After 17 h of magnetic

stirring, the reaction mixture is filtered,
concentrated and purified on silica to give compound 80
(5.65 g, 83%).
1H NMR (CDC13) δ8.77 (s, NH (isomer a)), 5.70 (dd, H-
3) , 2.64 (d, H-1β) .
Step 11.f: Preparation of methyl (3,6-di-O-acetyl-2-
azido-2,4-dideoxy-4-(prop-1-en-1-yl)-α-D-gluco-
pyranosyl) - (1-4) - (methyl 2-O-acetyl-3-O-benzyl-α-L-
idopyranosyluronate)-(L1-4)-(6-O-acetyl-3-O-benzyl-2-
benzyloxycarbonylamino-2-deoxy-α-D-glucopyranoside)
(No. 82)
Compound 80 (5.65 g, 12.3 mmol) obtained in
step 11.e and compound 81 (prepared according to J.C.
Jacquinet et al., Carbohydr. Res. 130 (1984), 221-241)
(11.54 g, 1.2 molar equivalents) are reacted according
to METHOD 2 to give, after purification, compound 82
(9.39 g, 71%).
Mass spectrum (ESI) m/z 1077.5 [(M+H)+] .
Step 11.g: Preparation of methyl (3,6-di-O-acetyl-2-
azido-2,4-dideoxy-4-(1,2-dihydroxypropyl)-α-D-
glucopyranosyl)-(1-4)-(methyl 2-O-acetyl-3-O-benzyl-α-
L-idopyranosyluronate)-(1-4)-(6-O-acetyl-3-O-benzyl-2-
benzyl oxycarbonyl amino-2-deoxy-α-D-glucopyranoside)
(No. 83)
N-Methylmorpholine N-oxide monohydrate (NMO)
(1.11 g, 20 molar equivalents) and 4% osmium tetraoxide
(OsO4) in water (8.35 ml, 1 molar equivalent) are added

to a solution of compound 82 (513. mg, 0.476 mmol)
obtained in step 11.f, in a 1:1 tetrahydrofuran-
dichloromethane mixture (8 ml). After stirring for
3 days at room temperature, a 1:1 dichloromethane-water
mixture is added as well as a 37.5% sodium hydrogen
sulphite (NaHSO3) solution, and the stirring is
maintained for an additional 30 min. The reaction
mixture is extracted with dichloromethane, purified on
silica and the fraction containing the starting
material is allowed to react under the above conditions
until it is completely consumed. After purification,
compound 83 (293 mg, 66%) is finally obtained.
Mass spectrum (ESI) m/z 1111.4 [ (M+H)+] .
Step 11.h: Preparation of methyl(3,6-di-O-acetyl-2-
azido-2,4-dideoxy-4-(5-methyl-2-phenyl-1,3-dioxolan-4-
yl) -a(-D-glucopyranosyl) - (1-4) - (methyl 2-O-acetyl-3-O-
benzyl-α-L-idopyranosyluronate)-(1-4)-(6-O-acetyl-3-O-
benzyl-2-benzyloxycarbonylamino-2-deoxy-α-D-
glucopyranoside) (No. 84)
CSA (21.6 mg, 0.2 mol equiv.) and benzylidene
dimethyl acetal (160 µl, 2.3 molar equivalents) are
added, under argon, to a solution of compound 83
(518 mg, 0.466 mmol) obtained in step 11.g, in
acetonitrile. After stirring for 1 h 30 min, the medium
is neutralized with triethylamine, concentrated to
dryness and then purified on silica to give compound 84
(454 mg, 74%) .

Mass spectrum (ESI) m/z 1199.5 [ (M+H)+] .
Step 11.1; Preparation of methyl (2-azido-2,4-dideoxy-
4- (5-methyl-2-phenyl-1, 3-dioxolan-4-yl) -α-D-gluco-
pyranosyl)-(1-4)-(3-O-benzyl-α-L-idopyranosyluronic
acid)-(1-4)-(3-O-benzyl-2-benzyloxycarbonylamino-2-
deoxy-α-D-glucopyranoside) (No. 85)
Compound 84 (215 mg, 0.18 mmol) obtained in
step 11.h is treated according to method 3. After 16 h
of magnetic stirring at room temperature, the medium is
diluted with methanol (12.5 ml) and then a 4N aqueous
sodium hydroxide solution (11.5 ml) is added at 0°C.
The mixture is stirred for 4 h at 0°C, acidified (pH 5)
with 6N hydrochloric acid and is extracted with
dichloromethane, washed with 5% Na2SO3 and finally with
saturated sodium chloride. After drying and
concentrating, the residue is purified on silica to
give compound 85 (157 mg, 86%) .
Mass spectrum (ESI) m/z 1017.3 [ (M+H)+] .
Step 11.j: Preparation of methyl (2-azido-2,4-dideoxy-
4-(5-methyl-2-phenyl-1,3-dioxolan-4-yl)-3,6-di-O-sodium
sulphonato-q-D-glucopyranosyl)-(1-4)-(sodium 3-O-
benzyl-2-O-sodium sulphonato-α-L-idopyranosyluronate)-
(1-4)-(3-O-benzyl-2-benzyloxycarbonylamino-2-deoxy-6-O-
sodium sulphonato-α-D-glucopyranoside)(No. 86)
Compound 85 (160 mg, 0.157 mmol) obtained in
step 11.i is treated according to METHOD 4 to give
compound 86 (215 mg, 95%) .

Mass spectrum (ESI) m/z 689.1 [ (M+H-3Na) 2-] .
Step 11.k; Preparation of methyl (2-amino-2,4-dideoxy-
4- (1, 2-dihydroxypropyl) -3 , 6-di-O-sodium sulphonato-α-D-
glucopyranosyl) - (1-4) - (sodium 2-O-sodium sulphonato-α-
L-idopyranosyluronate)-(1-4)-(2-amino-2-deoxy-6-O-
sodium sulphonato-α-D-glucopyranoslde) (No. 87)
Compound 86 (210 mg, 0.145 mmol) obtained in
step 11.j is treated according to method 5 without
acetic acid to give compound 87 (108 mg, 73%). If
necessary, the reaction is repeated several times until
the benzyl protons disappear completely by NMR.
Mass spectrum (ESI) m/z 996.1 [ (M+H-Na) ] .
Step 11.1: Preparation of methyl (2-N-sodium
sulphonato-2,4-dideoxy-4-(1,2-dihydroxypropyl)-3,6-di-
O-sodium sulphonato-α-D-glucopyranosyl)-(1-4)-(sodium
2-O-sodium sulphonato-α-L-idopyranosyluronate)-(1-4)-
(2-N-sodium sulphonato-2-deoxy-6-O-sodium sulphonato-α-
D-glucopyranoside) (No. 88)
Pyridine.SO3 complex (662 mg, 4.16 mmol) is
added, at 0°C, to a solution of compound 87 (106 mg,
0.104 mmol) obtained in step 11.k, in water (7 ml),
while the pH is kept at 9.3 with IN sodium hydroxide.
The temperature is then increased to room temperature,
the reaction medium is stirred for 16 h while the pH is
kept at 9.3, and is then purified on a Sephadex G-25
gel column equilibrated with a 0.2M sodium chloride
solution. After combining the fractions containing the

product and concentrating, the residue is purified by
the same Sephadex G-25 column eluted with water, to
give compound 88 (116 mg, 91%).
Mass spectrum (ESI) m/z 1199.8 [ (M+H-Na) -] .
Step 11.m: Preparation of methyl (2-N-sodium
sulphonato-2,4-dideoxy-4-formyl-3,6-di-O-sodium
sulphonato-α-D-glucopyranosyl)-(1-4)-(sodium 2-O-sodium
sulphonato-α-L-idopyranosyluronate)-(1-4)-(2-N-sodium
sulphonato-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranoside) (No. 89)
Sodium periodate (22.1 mg, 1.1 molar
equivalents) is added to a solution of compound 88
(115 mg, 94 µmol) obtained in step 11.1, in water
(1.9 ml). After 1 h of magnetic stirring, the reaction
medium is purified on a Sephadex G-15 gel column
equilibrated in water to give compound 89 (107 mg,
96%) .
PREPARATION 12:
Synthesis of (2-acetamido-3,4-di-O-benzyl-2-deoxy-6-O-
sodium sulphonato-α-D-glucopyranosyl)-(1-4)-(5-
(benzyloxy)-4-oxypiperidine-1-carboxylate of benzyl-3-
carboxylate of sodium (3S,4R,5R)) (No. 97)


Step 12.a: Preparation of (6-O-acetyl-2-azido-3,4-di-O-
benzyl-2-deoxy-α-D-glucopyranosyl) - (1-4) - ( (3S, 4R, 5R) -
dibenzyl 5-(benzyloxy)-4-oxypiperidine-1,3-
dicarboxylate) (No'. 94)
Compound 93 (prepared according to R. Verduyn
et al., Reel. Trav. Chim. the Netherlands, 109 (1990),
12, 591) (361 mg, 0.631 mmol) and compound 6 (200 mg,
0.421 mmol) obtained in step l.e are treated according
to method 2 to give, after purification, compound 94
(224 mg, 60%).
Mass spectrum (ESI) m/z 885.4 [ (M+H)+] .
Step 12.b: Preparation of (6-O-acetyl-2-acetamido-3,4-
di-O-benzyl-2-deoxy-α-D-glucopyranosyl)-(1-4)-
((3S,4R,5R)-dibenzyl 5-(benzyloxy)-4-oxypiperidine-1,3-
dicarboxylate) (No. 95)
Compound 94 (56.3 mg, 63.6 µmol) obtained in
step 12.a is treated as for the synthesis of 17
(step 3.f) to give compound 95 (54.5 mg, 95%).

Mass spectrum (ESI) m/z 901.2 [ (M+H)+] . Step 12.c: Preparation of (2-acetamido-3,4-di-O-benzyl-
2-deoxy-α-D-glucopyranosyl)-(1-4)- (3S,4R,5R)benzyl 5-
(benzyloxy)-4-oxypiperidine-1-carboxylate-3-carboxylic
acid (No. 96)
Compound 95 (101 mg, 50.9 jiimol) obtained in
step 12.b is treated according to METHOD 3. The residue
is used in the raw state in the next step.
Step 12.d: Preparation of (2-acetamido-3,4-di-O-benzyl-
2-deoxy-6-O-sodium sulphonato-α-D-glucopyranosyl)-(1-
4) -(5-(benzyloxy)-4-oxypiperidine-1-carboxylate of
benzyl-3-carboxylate of sodium (3S,4R,5R))(No. 97)
The crude compound 96 obtained in step 12.c
is treated according to METHOD 4, to give compound 97
(35 mg, 88% (2 steps)), which may be partially
esterified on the carboxylic acid functional group.
Mass spectrum (ESI) m/z 847.2 [(M-H)"].
PREPARATION 13:
Synthesis of (3R,4R,5R)-benzyl 3-(benzyloxy)-5-
[(benzyloxy)methyl]-4-hydroxypiperidine-1-carboxylate
(No. 100)

Step 13.a: Preparation of (4aR,8R,8aR)benzyl 8-
(benzyloxy)-2-phenyltetrahydro-4H-[1,3] dioxino[5,4-

c]pyridine-6(5H)carboxylate (No. 99)
Camphorsulphonic acid (31 mg, 0.2 mol. equiv.)
and then benzaldehyde dimethyl acetal (0.23 ml, 2.3 mol
equiv.) are added to a solution of compound 5 (250 mg,
0.67 mmol) in acetonitrile (13.4 ml). After 1 h of
magnetic stirring at room temperature, the reaction
medium is neutralized with triethylamine, concentrated
and purified on silica gel (15:85 ethyl acetate-
cyclohexane) to give compound 99 (281 mg, 91%).
Mass spectrum (ESI) m/z 482.2 [ (M+Na)+] .
Step 13.b: Preparation of (3R,4R,5R)-benzyl 3-
(benzyloxy)-5-[(benzyloxy)methyl]-4-hydroxypiperidine-
1-carboxylate (No. 100)
Triethylsilane (0.20 ml, 4 molar
equivalents), trifluoroacetic acid (0.09 ml, 4 molar
equivalents) and trif luoroacetic anhydride (3 µl,
0.07 molar equivalent) are successively added, at 0°C,
under argon, to a solution of compound 99 (141 mg,
0.31 mmol) obtained in step 13.a, in dichloromethane
(1.2 ml). The temperature is kept at 0°C for 5 min and
then the reaction medium is placed at room temperature
for 3.5 h. The reaction mixture is then neutralized
with an aqueous sodium hydrogen carbonate solution,
with water, and the organic phase is dried (Na2SO4) ,
filtered and concentrated under vacuum. The residue is
purified on silica gel to give compound 100 (76 mg,
54%) .

Mass spectrum (ESI) m/z 462.3 [ (M+H)+] .
PREPARATION 14:
Synthesis of (3-O-benzyl-2,4-di-O-sodium sulphonato-α-
L-idopyranosyluronate of sodium)-(1-4)-(2-acetamido-3-
O-benzyl-2-deoxy-6-O-sodium sulphonato-α-D-gluco-
pyranosyl)-(1-4)-(3-O-benzyl-2-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(2-acetamido-3-O-
benzyl-2-deoxy-6-O-sodium sulphonato-or-D-gluco-
pyranosyl)-(1-4)-((3R,4R,5R)-benzyl 3-(benzyloxy)-5-
[(benzyloxy)methyl]-4-oxypiperidine-1-carboxylate)
(No. 104)


Step 14.a: Preparation of (benzyl-2,4-di-O-acetyl-3-O-
benzyl-α-L-idopyranosyluronate)-(1-4)-(6-O-acetyl-2-
azido-3-O-benzyl-2-deoxy-α-D-glucopyranosyl) - (1-4) -
(benzyl 2-Q-acetyl-3-Q-benzyl-α-L-idopyranosyluronate)-
(1-4) - (6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-α-D-gluco-
pyranosyl)-((3R,4R,5R)-benzyl 3-(benzyloxy)-5-[(benzyl-
oxy)methyl]-4-oxypiperidine-1-carboxylate) (No. 101)
Compounds 15 (123 mg, 0.075 mmol) and 100
(69 mg, 0.149 mmol) are treated according to METHOD 2
to give, after purification, compound 101 (117 mg,

80%) . a/β ratio 55/45.
Mass spectrum (ESI) m/z 1962 [ (M+Na)+] .
Step 14.b: Preparation of (benzyl 2,4-di-O-acetyl-3-O-
benzyl-α-L-idopyranosyluronate)-(1-4)-(6-O-acetyl-2-
acetamido-3-O-benzyl-2-deoxy-α-D-glucopyranosyl) - (1-4) -
(benzyl 2-O-acetyl-3-O-benzyl-α-L-idopyranosyluronate)-
(1-4) - (6-O-acetyl-2-acetamido-3-O-benzyl-2-deoxy-α-D-
glucopyranosyl) - (1-4).- ( (3R,4R,5R) -benzyl 3- (benzyloxy) -
5-[(benzyloxy)methyl]-4-oxypiperidine-1-carboxylate)
(No. 102)
Compound 101 (117 mg, 60 µmol) is dissolved
in pyridine (1 ml) and then thioacetic acid (1 ml,
225 molar equivalents) is added at 0°C. The reaction
medium is stirred for 17 h at room temperature and is
then concentrated and purified on silica gel (4:96
ethanol-toluene) to give compound 102 (50 mg, 42%).
Mass spectrum (ESI) m/z 1971.9 [ (M+H)+] .
Step 14.c: Preparation of 3-O-benzyl-α-L-
idopyranosyluronic acid-(1-4)-(2-acetamido-3-O-benzyl-
2-deoxy-a?-D-glucopyranosyl) - (1-4) - (3-O-benzyl-α-L-
idopyranosyluronic acid)-(1-4)-(2-acetamido-3-O-benzyl-
2-deoxy-α-D.-glucopyranosyl) - (1-4) - ( (3R, 4R, 5R) -benzyl 3-
(benzyloxy)-5-[(benzyloxy)methyl]-4-oxypiperidine-1-
carboxylate) (No. 103)
Compound 102 (50 mg, 25 µmol) is treated
according to METHOD 3 to give the derivative 103.
Mass spectrum (ESI) m/z 1581.7 [ (M+H)+] .

Step 14.d: Preparation of (3-O-benzyl-2,4-di-O-sodium
sulphonato-α-L-idopyranosyluronate of sodium)-(1-4)-(2-
acetamido-3-O-benzyl-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranosyl)-(1-4)-(3-O-benzyl-2-O-sodium
sulphonato-α-L-idopyranosyluronate of sodium)-(1-4)-(2-
acetamido-3-O-benzyl-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranosyl)-(1-4)-((3R,4R,5R)-benzyl 3-(benzyloxy)-
5-[(benzyloxy)methyl]-4-oxypiperidine-1-carboxylate)
(No. 104)
Compound 103 is treated according to
METHOD 4, to give compound 104 (43 mg, 80%, (2 steps)).
Mass spectrum (ESI) m/z 2134.3 [(M-Na) -] .
PREPARATION 15:
Synthesis of (benzyl 2-O-acetyl-3-O-benzyl-4-O-phenyl-
propyl-α-L-idopyranosyluronate)-(1-4)-(6-O-acetyl-2-
azido-3-O-benzyl-2-deoxy-a, β-D-glucopyranose trichloro-
acetimidate) (No. 114)


Step 15.a: Preparation of (4,6-O-isopropylidene-3-O-
benzyl-2-O- (4-methoxy) benzyl-α-L-idopyranosyl) - (1-4) -
(1, 6-anhydro-2-azido-3-O-benzyl-2-deoxy-β-D-
glucopyranose) (No. 106)
NaH (6.93 g, 1.3 molar equivalents) and then
parα-methoxybenzyl chloride (24 ml, 1.6 molar
equivalents) are added, at 0°C and under argon, to a
solution of compound 105 (63.2 g, 111 mmol) (prepared
according to C.A.A. van Boeckel et al., J. Carbohydrate
Chemistry (1985) 4 (3), 293-321) in DMF (445 ml). After
2 h of magnetic stirring, methanol is added (9 ml) , the
reaction medium is concentrated under vacuum, the crude

reaction product is diluted with ethyl acetate, washed
with water, dried (Na2SO4) , filtered and concentrated.
The residue obtained is purified on silica (ethyl
acetate-cyclohexane 15:85) to give 106.
Mass spectrum (ESI) m/z 707.3 [(M+NH4)+].
Step 15.b: Preparation of (3-O-benzyl-2-O-(4-
methoxy)benzyl-α-L-idopyranosyl)-(1-4)-(1,6-anhydro-2-
azido-3-O-benzyl-2-deoxy-β-D-glucopyranose) (No. 107)
Compound 106 obtained in the preceding step
is exposed to acetic acid at 80% in water. After 15 h
of magnetic stirring, the reaction mixture is cooled
with ice, diluted (dichloromethane) and neutralized
with sodium hydrogen carbonate. The organic phase is
dried (Na2SO4) , filtered and concentrated. The residue
obtained is purified on silica (ethyl acetate-
cyclohexane 3:7) to give 107 (63.2 g, 88%, 2 steps).
Mass spectrum (ESI) m/z 672.3 [(M+Na)+].
Step 15.c: Preparation of (3-O-benzyl-6-O-tert-
butyldimethylsilyl-2-O-(4-methoxy)benzyl-α-L-
idopyranosyl)-(1-4)-(1,6-anhydro-2-azido-3-O-benzyl-2-
deoxy-p-D-glucopyranose) (No. 10 8)
Compound 107 (64.2 g) is dissolved in
dichloromethane (2 00 ml). Triethylamine (30.3 ml,
2.2 molar equivalents), 4-dimethylaminopyridine-
(1.21 g, 0.1 molar equivalent) and tert-
butyldimethylsilyl chloride (17.04 g, 1.1 molar
equivalents) are successively added at 0°C and under

argon. After 4 h of magnetic stirring, 10% tert-
butyldimethylsilyl chloride is added and after one
hour, the reaction medium is diluted with
dichloromethane, washed with water, dried (Na2SO4) ,
filtered and concentrated. The residue obtained is
purified on silica (ethyl acetate-cyclohexane 15:85) to
give 108.
Mass spectrum (ESI) m/z 786.3 [(M+Na)+] .
Step 15.d: Preparation of (3-O-benzyl-6-O-tert-
butyldimethylsilyl-2-O-(4-methoxy)benzyl-4-O-
phenylpropyl-α-L-idopyranosyl)-(1-4)-(1,6-anhydro-2-
azido-3-O-benzyl-2-deoxy-β-D-glucopyranose) (No. 109)
Phenylpropyl bromide (74 ml, 5 molar
equivalents) and then NaH (7 g, 1.5 molar equivalents)
are added, at 0°C and under argon, to a solution of
compound 108 in dimethylformamide (485 ml). After 5.5 h
of magnetic stirring, methanol is added (50 ml), the
reaction medium is concentrated under vacuum, the crude
reaction product is diluted with ethyl acetate, washed
with water, dried (Na2SO4) , filtered and concentrated.
The residue obtained is purified on silica (ethyl
acetate-cyclohexane 15:85) to give 109 (49.3 g, 58%,
2 steps).
Mass spectrum (ESI) m/z 904.3 [(M+Na)+].
Step 15.e-f: Preparation of (2-O-acetyl-3-O-benzyl-6-O-
tert-butyldimethylsilyl-4-O-phenylpropyl-α-L-
idopyranosyl)-(1-4)-(l,6-anhydro-2-azido-3-O-benzyl-2-

deoxy-β-D-glucopyranose) (No. 110)
Water (112 ml) is added to a solution of 109
(49.3 g, 55.9 mmol) in dichloromethane (2.2 1)
followed, at 0°C, by DDQ (19.03 g, 1.5 molar
equivalents). After stirring for 3 h at 0°C, a sodium
hydrogen carbonate solution is added. The organic phase
is dried (Na2SO4) , filtered and concentrated. The
residue obtained is dissolved in pyridine (335 ml) and
then acetic anhydride (28 ml) and 4-
dimethylaminopyridine (682 mg) are added. After 16 h of
magnetic stirring, the reaction mixture is concentrated
under vacuum and the residue obtained is purified on
silica (ethyl acetate-cyclohexane 15:85) to give 110
(34.4 g, 77%, 2 steps).
Mass spectrum (ESI) m/z 826.4 [(M+Na)+].
Step 15.d-h: Preparation of (benzyl 2-O-acetyl-3-O-
benzyl-4-O-phenylpropyl-α-L-idopyranosyluronate) - (1-4) -
(1,6-anhydro-2-azido-3-O-benzyl-2-deoxy-β-D-
glucopyranose) (No. Ill)
A 3.5M aqueous sulphuric acid solution
(45 ml) containing chromic anhydride (10 g) is added,
at 0°C, to a solution of 110 (25.17 g, 31.3 mmol) in
acetone (1.46 1). After 3 h of magnetic stirring at
0°C, the reaction medium is diluted with
dichloromethane, washed with water, dried, filtered and
concentrated to give a crude reaction product which is
used directly in the next step. The residue obtained

above is dissolved in dimethylformamide (23 0 ml) and
potassium hydrogen carbonate (16.7 g, 5 molar
equivalents) and benzyl bromide (39.8 ml, 10 molar
equivalents) are added. The reaction mixture is stirred
for 16 h at room temperature and is then diluted with
ethyl acetate, washed with water, dried, filtered,
concentrated and purified on silica gel (ethyl acetate-
toluene 1:4) to give compound 111 (22.6 g, 91%,
2 steps).
Mass spectrum (ESI) m/z 811.3 [(M+NH4)+].
Step 15.i: Preparation of (benzyl 2-O-acetyl-3-O-
benzyl-4-O-phenylpropyl-α-L-idopyranosyluronate)-(1-4)-
(1, 6-di-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a?, β-D-
glucopyranose) (No. 112)
Trifluoroacetic acid (TFA) (1.14 ml, 11 molar
equivalents) is added, at 0°C, to a solution of
compound 111 (1.11 g, 1.39 mmol) in acetic anhydride
(13.2 ml, 100 molar equivalents). After returning to
room temperature, the reaction mixture is stirred for
3.5 h and is then concentrated, coevaporated with
toluene and purified on silica gel (85:15 toluene-ethyl
acetate) to give compound 112 (1.15 g, 93%).
Mass spectrum (ESI) m/z 918.3 [ (M+Na)+] .
Step 15.j: Preparation of (benzyl 2-O-acetyl-3-O-
benzyl-4-O-phenylpropyl-α-L-idopyranosyluronate)-(1-4)-
(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-g,β-D-
glucopyranose) (No. 113)

Benzylamine (BnNH2) (5.25 ml, 38 molar
equivalents) is added, at 0°C, to a solution of
compound 112 (1.13 g, 1.2 6 mtnol) in diethyl ether
(51 ml). After stirring for 5 h 15 min at room
temperature, the medium is acidified with 1N HCl and is
then extracted with diethyl ether, dried (Na2SO4) ,
concentrated and purified on silica gel (35:65 ethyl
acetate-cyclohexane) to give 113 (0.97 g, 90%).
Mass spectrum (ESI) m/z 854.3 [ (M+H)+] .
Step 15.k: Preparation of (benzyl 2-O-acetyl-3-O-
benzyl-4-O-phenylpropyl-α-L-idopyranosyluronate) -(1-4)-
(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-a, β-D-
glucopyranose trichloroacetimidate) (No. 114)
Caesium carbonate (Cs2CO3) (0.583 g,1.6 molar
equivalents) and then trichloroacetonitrile (CCl3CN)
(0.56 ml, 5.0 molar equivalents) are added, under
argon, to a solution of compound 113 (0.95 g,
1.12 mmol) in dichloromethane . (21.2 ml). After stirring
for 35 min, the reaction mixture is filtered and then
concentrated. The residue is purified on silica gel
(25:75 ethyl acetate-cyclohexane) to give 114 (995 mg,
90%) .
Mass spectrum (ESI) m/z 1021.5 [ (M+Na)+] .
PREPARATION 16:
Synthesis of (benzyl 3-O-benzyl-4-O-phenylpropyl-2-O-
sodium sulphonato-α-L-idopyranosyluronate) - (1-4) - (2-
acetamido-3-O-benzyl-2-deoxy-6-O-sodium sulphonato-α-D-

glucopyranosyl)-(1-4)-(benzyl 3-O-benzyl-2-O-sodium
sulphonato-α-L-idopyranosyluronate)-(1-4)-(2-acetamido-
3-O-benzyl-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranosyl)-(1-4)-((3S,4R,5R)-dibenzyl 5-
(benzyloxy)-4-oxypiperidine-1,3-dicarboxylate)
(No. 122)



Step 16..a: Preparation of (benzyl 2-O-acetyl-3-O-
benzyl-4-O-phenylpropyl-α-L-idopyranosyluronate) -(1-4)-
(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-α-D-
glucopyranosyl)-(1-4)-(benzyl 2-O-acetyl-3-O-benzyl-α-
L-idopyranosyluronate)-(1-4)-(1,6-anhydro-2-azido-3-O-
benzyl-2-deoxy-β-D-glucopyranose) (No. 115)
Compound 114 (990 mg, 0.99 mmol) and compound
8 (1.15 g, 1.7 mmol) are treated according to method 2
to give, after purification, compound 115 (623 mg,
42%) .
Mass spectrum (ESI) m/z 1533.8 [(M+Na)+] .
Step 16.b: Preparation of (benzyl 2-O-acetyl-3-O-
benzyl-4-O-phenylpropyl-α-L-idopyranosyluronate)-(1-4)-
(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-α-D-
glucopyranosyl)-(1-4)-(benzyl 2-O-acetyl-3-O-benzyl-α-
L-idopyranosyluronate)-(1-4)-(1,6-di-O-acetyl-2-azido-
3-O-benzyl-2-deoxy-g,β-D-glucopyranose) (No. 116)
Compound 115 (590 mg, 0.39 mmol) is treated
as for the synthesis of compound 112 to give, after
purification on silica gel (7:3 cyclohexane-ethyl
acetate), 116 (609 mg, 97%).
Mass spectrum (ESI) m/z 1636.2 [ (M+Na)+] .
Step 16.c: Preparation of (benzyl 2-O-acetyl-3-O-
benzyl-4-O-phenylpropyl-α-L-idopyranosyluronate)-(1-4)-
(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-α-D-
glucopyranosyl)-(1-4)-(benzyl 2-O-acetyl-3-O-benzyl-α-
L-idopyranosyluronate)-(1-4)-(6-O-acetyl-2-azido-3-O-

benzyl-2-deoxy-g, β-D-glucopyranose) (No. 117)
Compound 116 (592 mg, 0.367 mmol) is treated
as for the synthesis of compound 113 to give, after
purification on silica gel (65:35 cyclohexane-ethyl
acetate), compound 117 (530 mg, 92%).
Mass spectrum (ESI) m/z 1593.9 [ (M+Na)+] .
Step 16.d: Preparation of (benzyl 2-O-acetyl-3-O-
benzyl-4-O-phenylpropyl-α-L-idopyranosyluronate)-(1-4)-
(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-α-D-
glucopyranosyl)-(1-4)-(benzyl 2-O-acetyl-3-O-benzyl-α-
L-idopyranosyluronate)-(1-4)-(6-O-acetyl-2-azido-3-O-
benzyl-2-deoxy-α, (3-D-glucopyranose
trichloroacetimidate) (No. 118)
Compound 117 (511 mg, 0.325 mmol) is treated
as for the synthesis of compound 114 to give, after
purification on silica gel (7:3 cyclohexane-ethyl
acetate), 118 (495 mg, 89%).
Elemental analysis calculated for C85H90Cl3N7O25 :
C, 59.49; H, 5.29; N, 5.71.
Found: C, 59.49; H, 5.50; N, 5.48.
Step 16.e: Preparation of (benzyl 2-O-acetyl-3-O-
benzyl-4-O-phenylpropyl-α-L-idopyranosyluronate)-(1-4)-
(6-O-acetyl-2-azido-3-O-benzyl-2-deoxy-α-D-
glucopyranosyl)-(1-4)-(benzyl 2-O-acetyl-3-O-benzyl-α-
L-idopyranosyluronate)-(1-4)-(6-O-acetyl-2-azido-3-O-
benzyl-2-deoxy-α-D-glucopyranosyl)-(1-4)-((3S,4R,5R)-
dibenzyl 5-(benzyloxy)-4-oxypiperidine-1,3-

dicarboxylate) (No. 119)
Compounds 118 (497 mg, 0.279 mmol) and 6
(255 mg, 0.536 mmol) are treated according to METHOD 2
to give, after purification, compound 119 (375 mg,
66%) .
Elemental analysis calculated for C111H117N7O30: C, 59.4 9;
H, 5.29; N, 5.71.
Found: C, 59.49; H, 5.50; N, 5.48.
Step 16.f: Preparation of (benzyl 2-O-acetyl-3-O-
benzyl-4-O-phenylpropyl-α-L-idopyranosyluronate)-(1-4)-
(6-O-acetyl-2-acetamido-3-O-benzyl-2-deoxy-α-D-
glucopyranosyl)-(1-4)-(benzyl 2-O-acetyl-3-O-benzyl-α-
L-idopyranosyluronate)-(1-4)-(6-O-acetyl-2-acetamido-3-
O-benzyl-2-deoxy-α-D-glucopyranosyl)-(1-4)-((3S,4R, 5R)-
dibenzyl 5-.(benzyloxy) -4-oxypiperidine-1, 3-
dicarboxylate) (No. 12 0)
Compound 119 (180 mg, 88.7 µmol) is dissolved
in pyridine (1.4 ml) and then thioacetic acid (1.4 ml,
225 molar equivalents) is added at 0°C. The reaction
medium is stirred for 17 h at room temperature and is
then concentrated and purified on silica gel (4:1
toluene-acetone) to give compound 120 (153 mg, 84%).
Mass spectrum (ESI) m/z 2084.8 [ (M+Na)+] .
Step 16.g: Preparation of (benzyl 3-O-benzyl-4-O-
phenylpropyl-α-L-idopyranosyluronate)-(1-4) -(2-
acetamido-3-O-benzyl-2-deoxy-α-D-glucopyranosyl)-(1-4)-
(benzyl 3-O-benzyl-α-L-idopyranosyluronate)-(1-4)-(2-

acetamido-3-O-benzyl-2-deoxy-α-D-glucopyranosyl)-(1-4)-
((3S,4R,5R)-dibenzyl 5-(benzyloxy)-4-oxypiperidine-1,3-
dicarboxylate) (No. 121)
Compound 120 (190 mg, 93.6 µmol) is treated
according to METHOD 3. The polyol obtained is dissolved
in dimethylformamide (4.4 ml), and potassium hydrogen
carbonate (85 mg, 10 molar equivalents) and benzyl
bromide (202 µl, 20 molar equivalents) are added at
0°C. The reaction mixture is stirred at room
temperature for 16 h and is then purified on an LH-2 0
column.
Purification on silica gel (ethyl acetate-
cyclohexane 2:3) makes it possible to obtain 121
(108 mg, 62% (2 steps)).
Mass spectrum (ESI) m/z 1884.2 [(M+Na)+] .
Step 16.h: Preparation of (benzyl 3-O-benzyl-4-O-
phenylpropyl-2-Q-sodium sulphonato-α-L-
idopyranosyluronate)-(1-4)-(2-acetamido-3-O-benzyl-2-
deoxy-6-O-sodium sulphonato-α-D-glucopyranosyl)-(1-4)-
(benzyl 3-O-benzyl-2-O-sodium sulphonato-α-L-
idopyranosyluronate)-(1-4)-(2-acetamido-3-O-benzyl-2-
deoxy-6-O-sodium sulphonato-α-D-glucopyranosyl)-(1-4)-
((3S,4R,5R)-dibenzyl 5-(benzyloxy)-4-oxypiperidine-1,3 -
dicarboxylate) (No. 122)
Compound 121 (41 mg, 21.6 µmol) is treated
according to METHOD 4 to give compound 122 (49 mg, 99%).
Mass spectrum (ESI) m/z 2301.0 [(M-H)-] .

The examples which follow illustrate the
preparation of compounds of the invention without
limiting it. The mass and NMR spectra confirm the
structures of the compounds obtained.
EXAMPLE 1:
Synthesis of (2 , 4-di-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(2-acetamido-2-
deoxy-6-O-sodium sulphonato-oc-D-glucopyranosyl) - (1-4) -
(2-O-sodium sulphonato-α-L-idopyranosyluronate of
sodium)-(1-4)-(2-acetamido-2-deoxy~6-O-sodium
sulphonato-α-D-glucopyranosyl)-(1-4)-(5-(hydroxy)-4-
oxypiperidine-3-carboxylate of sodium (3S,4R,5R))
(compound No. 20)

Compound 19 of PREPARATION 3 (50 mg,
24.02 µmol) is treated according to method 5 to give
compound 20. (26 mg, 72%) .
1H NMR (D20) δ5.23 (d, H-1 GlcII) , 5.13 (d, H-1
IdoUAIII) , 5.10 (d, H-1 IdoUAv) , 5.09 (d, H-1 GlcIV) ,
3.62, 3.04, 2.64, 2.47 (4m, 4H, H-2, H-2', H-6, H-6'
pipI) .

EXAMPLE 2;
Synthesis of (2-acetamido-2-deoxy-6-O-sodium
sulphonato-α-D-glucopyranosyl)-(1-4)-(5-(hydroxy)-4-
oxypiperidine-3-carboxylate of sodium (3S,4R,5R))
(compound No. 21)

Compound 97 of PREPARATION 12 is treated
according to method 5 to give compound 21.
1H NMR (D20) δ5.21 (d, H-1 GlcII) , 3.40, 3.40, 3.40,
3.10 (4m, 4H, H-2, H-2', H-6, H-6' pipI).
EXAMPLE 3;
Synthesis of (2,4-di-O-sodium sulphonato-α-L-
idopyrariosyluronate of sodium)-(1-4) -(2-acetamido-2-
deoxy-6-O-sodium sulphonato-α-D-glucopyranosyl)-(1-4)-
(5-(hydroxy)-4-oxypiperidine-3-carboxylate of sodium
(3S,4R,5R)) (compound No. 22)
?

Compound 22 was prepared in the same manner.
1H NMR (D2O) S 5.19 (d, H-1 GlC11) , 5.15 (d, H-1
IdoUAIII) , 3.27, 3.23, 3.09, 2.89 (4m, 4H, H-2, H-2', H-
6, H-6' pipI) .
EXAMPLE 4;
Synthesis of (2-acetamido-2-deoxy-6-O-sodium
sulphonato-α-D-glucopyranosyl)-(1-4)-(2-O-sodium
sulphonato-α-L-idopyranosyluronate of sodium)-(1-4)-(2-
acetamido-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranosyl) -(1-4)-(5-(hydroxy)-4-oxypiperidine-3-
carboxylate of sodium (3S,4R,5R)) (compound No. 23)

Compound 23 was prepared in the same manner.
1H NMR (D2O) δ 5.26 (d, H-1 GlcII), 5.14 (d, H-1
IdoUA111), 5.09 (d, H-1 GlcIV) .
EXAMPLE 5;
Synthesis of (2,4-di-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(2-N-sodium
sulphonato-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranosyl) -(1-4)-(2-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(2-N-sodium

sulphonato-2 -deoxy- 6-O- sodium sulphonato-α-D-
glucopyranosyl)-(1-4)-(5-(hydroxy)-4-oxypiperidine-3-
carboxylate of sodium (3S,4R,5R)) (compound No. 27)

Compound 2 6 of PREPARATION 4 (7.0 mg,
3.2 8 µmol) is treated according to method 5 to give
compound 27- (2.9 mg, 55%).
1H NMR (D2O) δ5.53 (d, H-1 GlC11) , 5.44 (d, H-1 GlcIV) ,
5.23 (d, H-1 IdoUAIII) , 5.21 (d, H-1 IdoUAv) , 3.09, 3.07,
2.58, 2.44, (4m, 4H, H-2, H-2', H-6, H-6' pipI).
EXAMPLE 6:
Synthesis of (3-O-methyl-2,4-di-O-sodium sulphonato-α-
L-idopyranosyluronate of sodium)-(1-4)-(3-O-methyl-2,6-
di-O-sodium sulphonato-α-D-glucopyranosyl)-(1-4)-(3-O-
methyl-2-O-sodium sulphonato-α-L-idopyranosyluronate of
sodium)-(1-4)-(3-O-methyl-2,6-di-O-sodium sulphonato-α-
D-glucopyranosyl) -(1-4)-(5-(hydroxy)-4-oxypiperidine-3-
carboxylate of sodium (3S,4R,5R)) (compound No. 47)


Compound 46 of PREPARATION 6 is treated
according to method 5 to give, after purification,
compound 47(5 mg, 57%).
Mass spectrum (ESI) m/z 1650.9 [(M-Na+H)"] .
EXAMPLE 7:
Synthesis of (2,4-di-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(2,6-di-O-sodium
sulphonato-α-D-glucopyranosyl)-(1-4)-(2-O-sodium
sulphonato-α-L-idopyranosyluronate of sodium)-(1-4)-
(2,6-di-O-sodium sulphonato-α-D-glucopyranosyl)-(1-4)-
(5-(hydroxy)-4-oxypiperidine-3-carboxylate of sodium
(3S,4R,5R)) (compound No. 69)

The crude compound 68 of PREPARATION 9 is
treated according to METHOD.5 to give 69 (3.8 mg, 25%,
two steps).
1H NMR (D20) 8 5.62 (d, H-1 GlcIV) , 5.52 (d, H-1 GlcII) ,
5.21 (d, H-1 IdoUAv) , 4.99 (d, H-1 IdoUAIII) .
EXAMPLE 8;
Synthesis of (4-O-propyl-2-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(2,6-di-O-sodium
sulphonato-α-D-glucopyranosyl)-(1-4)-(2-O-sodium
sulphonato-α-L-idopyranosyluronate of sodium)-(1-4)-

(2,6-di-O-sodium sulphonato-α-D-glucopyranosyl)-(1-4)-
(5-(hydroxy)-4-oxypiperidine-3-carboxylate of sodium
(3S,4R,5R)) (compound No. 74)

The crude compound 73 of PREPARATION 10 is
treated according to METHOD 5 to give 74 (6.0 mg, 62%,
two steps).
1H NMR (D2O) S 5.68 (d, H-1 GlcIV) , 5.55 (d, H-1 GlC11) ,
5.21 (d, H-1 IdoUAv) , 5.18 (d, H-1 IdoUAIII) .
EXAMPLE 9;
Synthesis of methyl (2,4-di-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(2-acetamido-2-
deoxy-6-O-sodium sulphonato-α-D-glucopyranosyl)-(1-4)-
(5-(hydroxy)-4-oxypiperidine-1-yl-3-carboxylate of
sodium (3S,4R,5R))-(1-4)-(2-N-sodium sulphonato-2,4-
dideoxy-4-methyl-3,6-di-O-sodium sulphonato-α-D-
glucopyranosyl) - (1-4) - (sodium 2-O-sodium sulphonato-α-
L-idopyranosyluronate)-(1-4)-(2-N-sodium sulphonato-2-
deoxy-6-O-sodium sulphonato-α-D-glucopyranoside)
(compound No. 90)


Sodium cyanoborohydride (4.4 mg, 2.4 mol
equiv.) is added, at 0°C, to a solution of compound 89
(24 mg, 26.9 µmol) of PREPARATION 11, and of compound
22 (61 mg, 1.9 molar equivalents) of EXAMPLE 3 in ..
phosphate buffer (pH 7). After stirring for 8 h at 0°C,
the reaction medium is placed at room temperature for
16 h and then successively purified on a Sephadex® G-25
gel column eluted with 0.2M sodium chloride followed by
the same Sephadex® G-25 column eluted with water to
give compound 90 (17 mg, 31%).
Mass spectrum (ESI) m/z 2029.3 [(M+H-Na)-] .
EXAMPLE 10:
Synthesis of methyl (2,4-di-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(2-acetamido-2-
deoxy-6-O-sodium sulphonato-oc-D-glucopyranosyl) - (1-4) -
(5-(hydroxy)-4-oxypiperidine-l~yl-3-carboxylate of
sodium (3S,4R,5R))-(1-4)-(2-N-sodium sulphonato-2,4-
dideoxy-4-methyl- 6 -O-sodium sulphonato-α-D-
glucopyranosyl) - (1-4) - (sodium 2-O-sodium sulphonato-α-
L-idopyranosyluronate)-(1-4)-(2-N-sodium sulphonato-2-
deoxy-6-O-sodium sulphonato-α-D-glucopyranoside)
(compound No. 91)


Compound 91 was prepared in the same manner.
Mass spectrum (ESI) m/z 1927.1 [ (M+H-Na) -] .
EXAMPLE 11:
Synthesis of methyl (5-(hydroxy-4-oxypiperidine-1-yl-3-
carboxylate of sodium (3S,4R,5R))-(1-4)-(2-N-sodium
sulphonato-2,4-dideoxy-4-methyl-3,6-di-O-sodium
sulphonato-α-D-glucopyranosyl)-(1-4)-(sodium 2-O-sodium
sulphonato-oc-L-idopyranosyluronate)-(1-4)-(2-N-sodium
sulphonato-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranoside) (compound No. 92)

Compound 92 was prepared in the same manner. .
Mass spectrum (ESI) m/z 1321.1 [(M+H-Na)"].
EXAMPLE 12:
Preparation of (2-acetamido-2-deoxy-6-O-sodium
sulphonato-α-D-glucopyranosyl)-(1-4)-(5-(hydroxy-4-

oxypiperidine-1-methyl-3-carboxylate of sodium
(3S,4R,5R)) (compound No. 98)

Compound 97 of PREPARATION 12 (20 mg,
21.3 µmol) is treated according to METHOD 5 in a 3:1:1
methanol-acetic acid-water mixture under a hydrogen
stream to give compound 98 (6.0 mg, 57%)
Mass spectrum (ESI) m/z 456.8 [ (M-H)"] .
EXAMPLE 13:
Synthesis of (2,4-di-O-sodium sulphonato-oc-L-
idopyranosyluronate of sodium)-(1-4)-(2-acetamido-2-
deoxy-6-O-sodium sulphonato-oc-D-glucopyranosyl) - (1-4) -
(2-O-sodium sulphonato-α-L-idopyranosyluronate of
sodium)-(1-4)-(2-acetamido-2-deoxy-6-O-sodium
sulphonato-α-D-glucopyranosyl)-(1,4)-(5-(hydroxy)-3-
hydroxymethyl-4-oxypiperidine-(3R,4R,5R))
(compound No. 123)


Compound 104 (41 mg, 19 µmol) is treated
according to method 5 to give compound 12 3 (10.7 mg,
38%) .
1H NMR (D2O) δ5.28 (d, H-1 GlcII) , 5.20 (d, H-1 IdoUAIII) ,
5.17 (d, H-1 IdoUAv) , 5.15 (d, H-1 GlcIV) , 3.21, 3.19,
2.68, 2.59 (4m, 4H, H-2, H-2', H-6, H-6' pipI).
EXAMPLE 14:
Synthesis of (4-O-phenylpropyl-2-O-sodium sulphonato-α-
L-idopyranosyluronate of sodium) -(1-4)-(2-acetamido-2-
deoxy-6-O-sodium sulphonato-α-D-glucopyranosyl)-(1-4)-
(2-O-sodium sulphonato-α-L-idopyranosyluronate of
sodium)-(1-4)-(2-acetamido-2-deoxy-6-O-sodium
sulphonato-α-D-glucopyranosyl)-(1,4)-(5-(hydroxy)-4-
oxypiperidine-3-carboxylase of sodium (3R,4R,5R))
(compound No. 124)


Compound 122 is treated according to method 5
to give compound 124.
1H NMR (D2O) 8 5.28 (d, H-1 GlcII)., 5.21 (d, H-1 IdoUAv) ,
5.16 (d, H-1 IdoUAIII 5.16 (d, H-1 GlcIV) , 3.23, 3.20,
2.93, 2.75 (4m, 4H, H-2, H-2', H-6, H-6' pipI).

WE CLAIM :
1. Compounds of general formula (I):

in which:
R represents a hydrogen atom, a hydroxyl radical, an
-OSO3- radical, an -O- (C1-C5) alkyl radical or an
-O-aralkyl radical;
Z represents a COO- radical or a hydroxyl radical;
X represents a hydroxyl radical or a saccharide unit of
formula A:
in which:
- R1 represents an oxygen atom, allowing A to bind
to the azasugar unit or to another saccharide
unit,
- R2 represents an -NH2 radical, an -NHCO (C2-C5)-
alkyl radical, an -NHCOaryl radical, an -NHSO3-
radical, a hydroxyl radical, an -O-(C1-C5)alkyl
radical, an -O-aralkyl radical or an -0SO3-

radical,
- R3 represents a hydroxyl radical, an -0SO3-
radical, an -O-(C1-C5) alkyl radical or an
-O-aralkyl radical,
- R4 represents a hydroxyl radical, an -0SO3-
radical, an -O-(C1-C5)alkyl radical, an -O-aralkyl
radical or a saccharide unit of formula B:

in which:
- R6 represents an oxygen atom, allowing B to
bind to another saccharide unit of formula A,
- R7 and R8 have the same definition as R3 as
defined above,
- R9 represents a hydroxyl group, an -0SO3"
radical, an -O- (C1-C5) alkyl radical, an
-O-aralkyl radical or a saccharide unit of
formula A as defined above,
- R5 has the same definition as R3 as defined
above;
Y represents a hydrogen atom, a (C1-C5)alkyl radical or
a saccharide unit of formula D


in which:
- R10, R12 and R13 have the same definitions as R5, R3
and R2 respectively as defined above,
- R11 represents:

• a (C1-C3) alkylene radical allowing D to
attach to the azasugar unit, or
• an oxygen atom allowing D to attach to
another saccharide unit,
- R14 represents an -O- (C1-C5) alkyl radical or a
radical of formula -O-E in which E represents a
radical of formula:

in which:
- R15 represents an -O- (C1-C5) alkyl radical, an
-O-aralkyl radical or a saccharide unit of
formula D in which R11 represents an oxygen
atom,
- R16 and R17 have the same definition as R3 as
defined above,
provided, however, that when X and R each represent a

hydroxyl radical, Y does not represent a hydrogen atom,
and it being understood that the number of saccharide
units of which the compound of formula (I) is composed
is between 1 and 10,
in free form or in the form of salts formed with a
pharmaceutically acceptable base or acid, and in the
form of solvates or hydrates.
2. Compounds as claimed in Claim 1, of
general formula (I):

in which:
R represents a hydrogen atom, a hydroxyl radical, an
-OSO3- radical, an -O-(C1-C5) alkyl radical or an
-O-aralkyl radical;
Z represents a COO- radical or a hydroxyl radical;
X represents a hydroxyl radical or a saccharide unit of
formula A:
in which:
- R1 represents an oxygen atom,

- R2 represents an -NHCOCH3 radical, an -NHSO3-
radical, an -0SO3- radical,
- R3 represents a hydroxyl radical or an
-O-(C1-C5) alkyl radical,
- R4 represents a hydroxyl radical, an -O-aralkyl
radical or a saccharide unit of formula B:

in which:
- R6 represents an oxygen atom,
- R7 represents an -0SO3- radical,
- R8 represents a hydroxyl radical, an
-O-(C1-C5) alkyl radical or an -O-aralkyl
radical,
- R9 represents an -0SO3- radical, an
-O-aralkyl radical, an -O-(C1-C5)alkyl radical
or a saccharide unit of formula A as defined
above,
- R5 represents an -0SO3- radical;
Y represents a hydrogen atom or a saccharide unit of
formula D:
in which:

- R10 has the same definition as R5 as defined
above,
- R12 represents a hydroxyl radical or an -OSO3-
radical,
- R13 represents an -NHSO3- radical,
- R11 represents a methylene radical linked to an
azasugar unit or an oxygen atom linked to E,
- R14 an -OCH3 radical or a radical of formula -O-E
in which E represents a radical of formula:

in which:
- R15 represents a D unit in which R11
represents an oxygen atom allowing E to be
linked to D,
- R16 represents an -OSO3- radical,
- R17 represents a hydroxyl radical,
it being understood that the number of saccharide units
of which the compound of formula (I) is composed is
between 2 and 10,
in free form or in the form of salts with a
pharmaceutically acceptable base or acid, and in the
form of solvates or hydrates.
3. Compounds of general formula (I) as
claimed in either of Claims 1 and 2, in which Y is a

hydrogen atom and Z a COO- radical.
4. Compounds as claimed in any one of
Claims 1 to 3, chosen from:
• (2,4-di-O-sodium sulphonato-α-L-idopyranosyl-
uronate of sodium)-(1-4)-(2-acetamido-2-deoxy-6-
O-sodium sulphonato-α-D-glucopyranosyl)-(1-4)-
(2-O-sodium sulphonato-α-L-idopyranosyluronate
of sodium)-(1-4)-(2-acetamido-2-deoxy-6-O-sodium
sulphonato-α-D-glucopyranosyl)-(1-4)-(5-(hydroxy
)-4-oxypiperidine-3-carboxylate of sodium (3S,
4R, 5R))
• (2,4-di-O-sodium sulphonato-α-L-idopyranosyl-
uronate of sodium)-(1-4)-(2-N-sodium sulphonato-
2-deoxy-6-O-sodium sulphonato-α-D-gluco-
pyranosyl) - (1-4)-(2-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(2-N-sodium
sulphonato-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranosyl) -(1-4)-(5-(hydroxy)-4-oxy-
piperidine-3-carboxylate of sodium (3S, 4R, 5R))
• (3-O-methyl-2,4-di-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(3-O-
methyl-2,6-di-O-sodium sulphonato-α-D-gluco-
pyranosyl) -(1-4)-(3-O-methyl-2-O-sodium
sulphonato-α-L-idopyranosyluronate of sodium)-
(1-4)-(3-O-methyl-2,6-di-O-sodium sulphonato-α-
D-glucopyranosyl) -(1-4)-(5-(hydroxy)-4-oxy-
piperidine-3-carboxylate of sodium (3S, 4R, 5R))

(2,4-di-O-sodium sulphonato-α-L-idopyranosyl-
uronate of sodium)-(1-4)-(2,6)-di-O-sodium
sulphonato-α-D-glucopyranosyl)-(1-4)-(2-O-sodium
sulphonato-α-L-idopyranosyluronate of sodium)-
(1-4)-(2,6-di-O-sodium sulphonato-α-D-gluco-
pyranosyl)-(1-4)-(5-(hydroxy)-4-oxypiperidine-3-
carboxylate of sodium (3S, 4R, 5R))
(4-O-propy1-2-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(2,6-di-O-
sodium sulphonato-α-D-glucopyransyl-(1-4)-(2-O-
sodium sulphonato-α-L-idopyranosyluronate of
sodium)-(1-4)-(2,6-di-O-sodium sulphonato-α-D-
glucopyranosyl)-(1-4)-(5-(hydroxy)-4-
oxypiperidine-3-carboxylate of sodium (3S, 4R,
5R))
(2,4-di-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(2-
acetamido-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranosyl)-(1-4)-(2-O-sodium sulphonato-α-
L-idopyranosyluronate of sodium)-(1-4)-(2-
acetamido-2-deoxy-6-O-sodium sulphonato-cx-D-
glucopyranosyl)-(1-4)-(3-(hydroxy)-5-
hydroxymethyl-4-oxypiperidine (3R, 4R, 5R))
(4-O-phenylpropyl-2-O-sodium sulphonato-α-L-
idopyranosyluronate of sodium)-(1-4)-(2-
acetamido-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranosyl)-(1-4)-(2-O-sodium sulphonato-α-

L-idopyranosyluronate of sodium)-(1-4)-(2-
acetamido-2-deoxy-6-O-sodium sulphonato-α-D-
glucopyranosyl)-(1-4)-(5-(hydroxy)-4-oxy-
piperidine-3-carboxylate of sodium (3S, 4R,
5R)) .
5. Pharmaceutical compositions containing,
as active ingredient, a compound of general formula (I)
as claimed in any one of Claims 1 to 4, optionally
combined with one or more inert and appropriate
excipients.
6. Pharmaceutical composition as claimed in
Claim 5, capable of being used in the treatment of
carcinomas having a high degree of vascularization,
cardiovascular diseases, chronic inflammatory diseases
and macular degeneration.


(57) Abstract: The invention concerns heparanase inhibiting compounds of general formula (I) wherein R represents a hydrogen
atom, an hydroxyl radical, a -OSO3- radical, a -O-(C1-C5 alkyl radical or an -O- aralky) radival; Z represents a COO radical or a
hydroxy) radical; X represents -OH or a saccharide unit of formula A, Y represents H, C1-C3 alkyl or a saccharide unit of formula
D; in fire form or in the form of pharmaceutically aceeptable salts formed with a base or an acid as well as in the form of solvates
or hydrates. The derivatives of the invention are useful as medicines.

Documents:

03268-kolnp-2006 abstract.pdf

03268-kolnp-2006 assignment.pdf

03268-kolnp-2006 claims.pdf

03268-kolnp-2006 correspondence other.pdf

03268-kolnp-2006 description(complete).pdf

03268-kolnp-2006 form-1.pdf

03268-kolnp-2006 form-3.pdf

03268-kolnp-2006 form-5.pdf

03268-kolnp-2006 international publication.pdf

03268-kolnp-2006 international search authority report.pdf

03268-kolnp-2006 pct other document.pdf

03268-kolnp-2006-assignment-1.1.pdf

03268-kolnp-2006-correspondence-1.1.pdf

3268-KOLNP-2006-(28-09-2011)-CORRESPONDENCE.pdf

3268-KOLNP-2006-(28-09-2011)-ENGLISH TRANSLATION.pdf

3268-KOLNP-2006-(28-09-2011)-PETITION UNDER RULE 137.pdf

3268-KOLNP-2006-ABSTRACT-1.1.pdf

3268-KOLNP-2006-AMANDED CLAIMS.pdf

3268-KOLNP-2006-ASSIGNMENT.pdf

3268-KOLNP-2006-CORRESPONDENCE-1.1.pdf

3268-KOLNP-2006-CORRESPONDENCE.pdf

3268-KOLNP-2006-DESCRIPTION (COMPLETE)-1.1.pdf

3268-KOLNP-2006-ENGLISH TRANSLATION.pdf

3268-KOLNP-2006-EXAMINATION REPORT.pdf

3268-KOLNP-2006-FORM 1-1.1.pdf

3268-KOLNP-2006-FORM 18-1.1.pdf

3268-kolnp-2006-form 18.pdf

3268-KOLNP-2006-FORM 2.pdf

3268-KOLNP-2006-FORM 3.pdf

3268-KOLNP-2006-FORM 5.pdf

3268-KOLNP-2006-GPA.pdf

3268-KOLNP-2006-GRANTED-ABSTRACT.pdf

3268-KOLNP-2006-GRANTED-CLAIMS.pdf

3268-KOLNP-2006-GRANTED-DESCRIPTION (COMPLETE).pdf

3268-KOLNP-2006-GRANTED-FORM 1.pdf

3268-KOLNP-2006-GRANTED-FORM 2.pdf

3268-KOLNP-2006-GRANTED-SPECIFICATION.pdf

3268-KOLNP-2006-OTHERS.pdf

3268-KOLNP-2006-OTHERS1.1.pdf

3268-KOLNP-2006-PETITION UNDER RULE 137.pdf

3268-KOLNP-2006-REPLY TO EXAMINATION REPORT.pdf

3268-KOLNP-2006-TRANSLATED COPY OF PRIORITY DOCUMENT.pdf


Patent Number 253108
Indian Patent Application Number 3268/KOLNP/2006
PG Journal Number 26/2012
Publication Date 29-Jun-2012
Grant Date 26-Jun-2012
Date of Filing 08-Nov-2006
Name of Patentee SANOFI-AVENTIS
Applicant Address 174, AVENUE DE FRANCE FR-75013 PARIS FRANCE
Inventors:
# Inventor's Name Inventor's Address
1 PETITOU MAURICE 65 RUE DU JAVELOT FR-75645 PARIS CEDEX 13 FRANCE
2 DRIGUEZ PIERRE ALEXANDRE 16 RUE AUGUSTE DIDE FR-31500 TOULOUSE FRANCE
PCT International Classification Number C07H 3/06,A61K31/702
PCT International Application Number PCT/FR2005/001851
PCT International Filing date 2005-07-20
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 04/08,160 2004-07-23 France