FGF RECEPTOR-ACTIVATING 3-O-ALKYL OLIGOSACCHARIDES,
PREPARATION THEREOF AND THERAPEUTIC USE THEREOF
The present invention relates to 3-O-alkyl oligosaccharides which are agonists of
the FGFs/FGFRs system, to the preparation thereof and to the therapeutic use thereof.
Angiogenesis is a process of generation of new blood capillaries. During the
blockage of a blood vessel, angiogenesis, associated with arteriogenesis (dilation of the
capillaries), improves the revascularization of the blocked area. It has been shown in vitro
and in vivo that several growth factors, such as Vascular Endothelial Growth Factors
(VEGFs) and Fibroblast Growth Factors (FGFs), stimulate the neovascularisation process.
FGFs are a family of 23 members. FGF2 (or basic FGF) is an 18 kDa protein.
FGF2 induces, in endothelial cells in culture, their proliferation, their migration and the
production of proteases. In vivo, FGF2 promotes neovascularisation phenomena. FGF2
interacts with endothelial cells via two classes of receptors, high-affinity receptor tyrosine
kinases (FGFRs) and low-affinity receptors of heparan sulphate proteoglycan (HSPG)
type.
It is known that cell surface receptor tyrosine kinases associate in dimeric form
with a complex made up of two ligand molecules and one heparan sulphate molecule. The
formation of this complex makes it possible to trigger a cascade of intracellular signals
resulting in activation of cell proliferation and migration, which are two key processes
involved in angiogenesis.
Thus, FGF2 and its receptors represent very pertinent targets for therapies aimed
at activating or inhibiting angiogenesis processes.
We have now found novel synthetic 3-O-alkyl oligosaccharide compounds capable
of facilitating the formation of the FGF/FGFR complex and of promoting the formation of
new vessels in vitro and in vivo.
A subject of the present invention is novel oligosaccharide compounds
corres onding to formula (I):
in which:
- the wavy line denotes a bond located either below or above the plane of the
pyranose ring of the saccharide unit,
- R represents an -O-alkyl group, in which said alkyl group contains from 1 to 16
carbon atoms and is optionally substituted with one or more (for example 1 or 2) groups,
which may be identical or different, chosen from aryl and cycloalkyl groups,
- R2 represents a hydroxyl group or an -O-alkyl group,
- R3, R5, R6, R7 and R8, which may be identical to or different from one another,
represent either an -OS0 3 group or a hydroxyl group,
- R4 represents either an -NH-CO-alkyl group or an -O-alkyl group,
- R represents an -O-alkyl group, and
- n and m, which may be identical to or different from one another, represent
integers equal to 0 or 1.
In the context of the present invention, and unless otherwise mentioned in the text:
- the term "alkyl group" is intended to mean: a linear or branched saturated
aliphatic group advantageously comprising between 1 and 6 carbon atoms. By way of
examples, mention may be made of methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertbutyl,
pentyl, etc. groups;
- the term "cycloalkyl group" is intended to mean: a cyclic alkyl group comprising
from 3 to 6 carbon atoms, for example a cyclopentyl or cyclohexyl group;
- the term "aryl group" is intended to mean: a cyclic aromatic group comprising
between 5 and 10 carbon atoms, such as a phenyl group. Such an aryl group is optionally
substituted with one or more groups such as halogen atoms and alkyl, alkoxy, thioalkyl,
trifluoromethyl and phenyl groups.
The oligosaccharides according to the invention are synthetic in nature, in the
sense that they are compounds obtained by total synthesis from intermediate synthons,
as will be described in detail in the text hereinbelow. In this respect, they differ from
oligosaccharides obtained by depolymerisation or isolation from complex mixtures of
polysaccharides, such as heparins or low-molecular-weight heparins. In particular, the
compounds according to the invention have a well-defined structure resulting from their
chemical synthesis and are in the form of pure oligosaccharides, i.e. they are free of other
oligosaccharide entities.
The invention encompasses the compounds of formula (I) in acid form or in the
form of any one of the pharmaceutically acceptable salts thereof. In the acid form,
the -COO and -S0 3 functions are, respectively, in -COOH and -S0 3H form.
The expression "pharmaceutically acceptable salt of the compounds of the
invention" is intended to mean a compound in which one or more of the -COO and/or -
S0 3 functions are ionically bonded to a pharmaceutically acceptable cation. The preferred
salts according to the invention are those in which the cation is chosen from alkaline metal
cations, in particular the Na+ cation.
The compounds of formula (I) according to the invention also comprise those in
which one or more hydrogen or carbon atoms have been replaced with the radioactive
isotope thereof, for example tritium or carbon 4C. Such labelled compounds are of use in
research, metabolism or pharmacokinetic studies, as ligands in biochemical tests.
The oligosaccharides according to the invention stand out from those previously
known in that:
- the iduronic acids are substituted in position 3 with an alkoxy group, and
- the glucosamine units are substituted in position 2 with an acyl group
(-NH-CO-alkyl) or with an alkoxy group, and also with an alkoxy group in position 3.
The compounds according to the invention are advantageously octasaccharides,
i.e. compounds of formula (I) in which n = 1 and m = 0 or else n = 0 and m = 1.
Among the compounds of formula (I) which are subjects of the invention, mention
may be made of a subgroup of compounds in which R represents an -O-alkyl group,
where said alkyl group contains from 1 to 8 carbon atoms, advantageously from 1 to 5
carbon atoms (for example an -O-methyl or -O-pentyl group), and is optionally substituted
with 1 or 2 groups, which may be identical or different, chosen from aryl groups (such as
phenyl).
Among the compounds of formula (I) which are subjects of the invention, mention
may be made of another subgroup of compounds in which R2 represents a hydroxyl group
or an -O-alkyl group, where said alkyl group comprises from 1 to 4 carbon atoms.
Advantageously, the compounds of formula (I) according to the invention are such
that R2 represents a hydroxyl group.
Among the compounds of formula (I) which are subjects of the invention, mention
may be made of another subgroup of compounds in which R3, R5, R6, R7 and R8, which
may be identical to or different from one another, represent either an -OS0 3 group or a
hydroxyl group, on the condition that at least one group among R3, R5, R6, R7 and R8
represents an -OS0 3 group.
Another subgroup of compounds of formula (I) is such that at least one of the
groups R3, R5, R6, R 7 and R8 represents an -OS0 3 group and at least one of the groups
R3, R5, R6, R 7 and R8 represents a hydroxyl group.
Another subgroup of compounds of formula (I) is such that R3, R5, R6, R7 and R8 all
represent -OS0 3 groups.
Another subgroup of compounds of formula (I) is such that R3, R5 and R6 represent
-OS0 3 groups and R7 and R8 represent hydroxyl groups.
Among the compounds of formula (I) which are subjects of the invention, mention
may be made of another subgroup of compounds in which R4 represents an -NH-CO-alkyl
group, where said alkyl group comprises from 1 to 4 carbon atoms, for example a methyl,
propyl or isobutyl group.
Among the compounds of formula (I) which are subjects of the invention, mention
may be made of another subgroup of compounds in which R4 represents an -O-alkyI
group, where said alkyl group comprises from 1 to 4 carbon atoms, for example a butyl
group.
Among the compounds of formula (I) which are subjects of the invention, mention
may be made of another subgroup of compounds in which R represents an O-alkyI group,
where said alkyl group comprises from 1 to 4 carbon atoms.
Advantageously, the compounds of formula (I) according to the invention are such
that R represents a methoxy group.
Other subgroups of oligosaccharides according to the invention may have several
of the characteristics set out above for each of the subgroups previously defined.
Thus, another subgroup of oligosaccharides according to the invention may consist
of octasaccharides of formula (I), in which:
- n = 0 and m = 1,
- R represents an -O-alkyI group, in which said alkyl group contains from 1 to 5
carbon atoms (for example an -O-methyl or -O-pentyl group), and is optionally substituted
with 1 or 2 groups, which may be identical or different, chosen from aryl groups (such as
phenyl),
- R2 represents a hydroxyl group or an -O-alkyI group, in which said alkyl group
comprises from 1 to 4 carbon atoms,
- R5, R6, R7 and R8, which may be identical to or different from one another,
represent either an OS0 3 group or a hydroxyl group, on the condition that at least one of
the groups R5, R6, R 7 and R8 represents an -OSO3 group,
- R4 represents either an -NH-CO-alkyl group or an -O-alkyI group, where said
alkyl group comprises from 1 to 4 carbon atoms, and
- R represents an -O-alkyI group, in which said alkyl group comprises from 1 to 4
carbon atoms.
Such octasaccharides correspond to formula ( ) below:
Another subgroup of octasaccharides according to the invention consist of
compounds of formula (I), in which:
- n = 0 and m = 1,
- R represents an -O-methyl, -O-pentyl or -O-pentylphenyl group,
- R2 represents a hydroxyl group,
- R5, R6, R7 and R8, which may be identical to or different from one another,
represent either an -OS0 3 group or a hydroxyl group, on the condition that at least one of
the groups R5, R6, R 7 and R8 represents an -OS0 3 group,
- R4 represents either an -NH-CO-alkyl or an -O-alkyl group, where said alkyl group
comprises from 1 to 4 carbon atoms, and
- R represents an -O-alkyl group, in which said alkyl group comprises from 1 to 4
carbon atoms.
Advantageously, another subgroup of octasaccharides according to the invention
consists of compounds of formula (I), in which:
- n = 0 and m = 1,
- R represents an -O-methyl, -O-pentyl or -O-pentylphenyl group,
- R2 represents a hydroxyl group,
- R5, R6, R7 and R8, which may be identical to or different from one another,
represent either an -OS0 3 group or a hydroxyl group, on the condition that at least one of
the groups R5, R6, R7 and R8 represents an -OS0 3 group,
- R4 is chosen from the groups -NH-CO-methyl, -NH-CO-propyl, -NH-CO-isobutyl
and -O-butyl, and
- R represents an -O-methyl group.
Among the octasaccharides defined previously, mention may in particular be made
of those in which at least one of the groups R5, R6, R7 and R8 represents an -OS0 3 group
and at least one of the groups R5, R6, R7 and R8 represents a hydroxyl group.
Among the compounds of the invention, mention may in particular be made of the
following octasaccharides:
- methyl (sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-
( 1 4)-(2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a -D-glucopyranosyl)-
( 1 4)-[(sodium 3-0-methyl-2-0-sodium sulphonato-a -L-idopyranosyluronate)-(1 4)-(2-
acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a-D-glucopyranosyl)-(1 4)]2-
(sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-(1 4)-2-
acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a-D-glucopyranoside (No. 1);
- pentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-
( 1 4)-(2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a-D-glucopyranosyl)-
( 1 4)-[(sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-(1 4)-(2-
acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a-D-glucopyranosyl)-(1 4)]2-
(sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-(1 4)-2-
acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-p-D-glucopyranoside (No. 2);
- pentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-
( 1 4)-(2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a-Dglucopyranosyl)-(
1 4)-[(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1®4)-(2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-a-D-glucopyranosyl)-(1 4)]2-(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1®4)-2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-p-D-glucopyranoside (No. 3);
- 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1 4)-(2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a-
D-glucopyranosyl)-(1 4)-[(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1 4)-(2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a-
D-glucopyranosyl)-(1 4)]2-(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1 4)-2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-b-
D-glucopyranoside (No. 4);
- 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1®4)-(2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-a-D-glucopyranosyl)-(1 4)-[(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1®4)-(2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-a-D-glucopyranosyl)-(1 4)] 2-(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1 4)-2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-p-D-glucopyranoside (No. 5);
- 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1®4)-(2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-a-D-glucopyranosyl)-(1 4)-(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1®4)-(2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-a-D-glucopyranosyl)-(1 4)-(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1®4)-(2-butanoylamino-2-deoxy-3-0-methyl-a-D-glucopyranoside)-
( 1 4)-(sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-(1 4)-2-
(butanoylamino)-2-deoxy-3-0-methyl-p-D-glucopyranoside (No. 6);
- 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1®4)-(2-[(3-methylbutanoyl)amino]-2-deoxy-3-0-methyl-6-0-
sodium sulphonato-a-D-glucopyranosyl)-(1 4)-(sodium 3-0-methyl-2-0-sodium
sulphonato-a-L-idopyranosyluronate)-(1®-4)-(2-[(3-methylbutanoyl)amino]-2-deoxy-3-0-
methyl-6-O-sodium sulphonato-a-D-glucopyranosyl)-(1 4)-(sodium 3-0-methyl-2-0-
sodium sulphonato-a-L-idopyranosyluronate)-(1®4)-(2-[(3-methylbutanoyl)amino]-2-
deoxy-3-0-methyl-a-D-glucopyranoside)-(1 4)-(sodium 3-0-methyl-2-0-sodium
sulphonato-a-L-idopyranosyluronate)-(1®-4)-2-[(3-methylbutanoyl)amino]-2-deoxy-3-0-
methyl-p-D-glucopyranoside (No. 7);
- 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1 4)-(2-0-butyl-3-0-methyl-6-0-sodium sulphonato-a-Dglucopyranosyl)-(
1 4)-(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1 4)-(2-0-butyl-3-0-methyl-6-0-sodium sulphonato-a-Dglucopyranosyl)-(
1 4)-(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1 4)-(2-0-butyl-3-0-methyl-a-D-glucopyranosyl)-(1 4)-(sodium 3-
O-methyl-2-O-sodium sulphonato-a -L-idopyranosyluronate)-(1 4)-2-0-butyl-3-0-methyl-
b-D-glucopyranoside (No. 8); and
- 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a -Lidopyranosyluronate)-(
1 4)-(2-0-butyl-3-0-methyl-6-0-sodium sulphonato-a -Dglucopyranosyl)-(
1 4)-(sodium 3-0-methyl-2-0-sodium sulphonato-a -Lidopyranosyluronate)-(
1 4)-(2-0-butyl-3-0-methyl-6-0-sodium sulphonato-a -Dglucopyranosyl)-(
1 4)-(sodium 3-0-methyl-2-0-sodium sulphonato-a -Lidopyranosyluronate)-(
1 4)-(2-0-butyl-3-0-methyl-a -D-glucopyranosyl)-(1 4)-(sodium 3-
O-methyl-2-O-sodium sulphonato-a -L-idopyranosyluronate)-(1 4)-2-0-butyl-3-0-methyla-
D-glucopyranoside (No. 9).
In its principle, the process for preparing the compounds according to the invention
uses di- or oligosaccharide basic synthons prepared as previously reported in the
literature. Reference will be made in particular to the patents or patent applications
EP 0 300 099, EP 0 529 715, EP 0 621 282 and EP 0 649 854, and also to the publication
by C. Van Boeckel and M. Petitou published in Angew. Chem. Int. Ed. Engl., 1993, 32,
1671-1690. These synthons are then coupled to one another so as to provide an entirely
protected equivalent of an oligosaccharide according to the invention. This protected
equivalent is then converted into a compound according to the invention. In the coupling
reactions mentioned above, a "donor" di- or oligosaccharide, activated on its anomeric
carbon, reacts with an "acceptor" di- or oligosaccharide, bearing a free hydroxyl.
The specific synthesis schemes will be described in the detailed examples which
follow.
The present invention therefore relates to a process for preparing the
oligosaccharides of formula (I), characterized in that:
- in a first phase, a fully protected equivalent of the desired oligosaccharide (I) is
synthesized, comprising in position 2 of the glucosamine units either an amine function
precursor (carbamate or azide for example), or an alkoxy group,
- in a second phase, the positions that are to comprise sulphate groups on the final
molecule are deprotected and then O-sulphated,
- in a third phase, the whole of the compound is deprotected, and
- in a fourth phase, if necessary, the N-acyl groups are introduced (introduction of
R4 groups of acyl type).
The synthesis of the fully protected equivalent of the desired oligosaccharide (I) is
carried out according to reactions that are well known to those skilled in the art, using
methods for the synthesis of oligosaccharides (for example, G.J. Boons, Tetrahedron
(1996), 52, 1095-1 121 and patent applications WO 98/03554 and WO 99/36443), in which
a glycosidic bond-donating oligosaccharide is coupled with a glycosidic bond-accepting
oligosaccharide to give another oligosaccharide of which the size is equal to the sum of
the sizes of the two reactive entities. This sequence is repeated until the compound of
formula (I) is obtained, optionally in protected form. The nature and profile of the charge of
the desired final compound determine the nature of the chemical entities used in the
various steps of the synthesis, according to the rules well known to those skilled in the art.
Reference may be made, for example, to C. Van Boeckel and M. Petitou, Angew. Chem.
Int. Ed. Engl. (1993), 32, 1671-1690 or else to H. Paulsen, "Advances in selective
chemical syntheses of complex oligosaccharides", Angew. Chem. Int. Ed. Engl. (1982), 2 ,
155-173.
The compounds of the invention may naturally be prepared using various
strategies known to those skilled in the art of oligosaccharide synthesis. The process
described above is the preferred process of the invention. However, the compounds of
formula (I) can be prepared via other well-known methods of sugar chemistry, described,
for example, in "Monosaccharides, Their chemistry and their roles in natural products",
P.M. Collins and R.J. Ferrier, J. Wiley & Sons (1995) and by G.J. Boons in Tetrahedron
(1996), 52, 1095-1 121 .
The protecting groups, used in the process for preparing the compounds of
formula (I), are those that make it possible firstly to protect a reactive function such as a
hydroxyl or an amine during a synthesis, and secondly to regenerate the intact reactive
function at the end of the synthesis. In the present application, these protecting groups are
denoted Pg, Pg' and Pg". The protecting groups commonly used in sugar chemistry, as
described, for example, in "Protective Groups in Organic Synthesis", Green e al., 3rd
Edition (John Wiley & Sons, Inc., New YorkJ, are used to carry out the process according
to the invention. The protecting groups are chosen, for example, from acetyl, azide,
benzoyl, benzyl, substituted benzyl, benzyl carbamate, isopropylidene, levulinoyl, methyl,
tetrahydropyranyl, fe/t-butyldimethylsilyl (tBDMS) and ie/f-butyldiphenylsilyl (tBDPS)
groups.
Activating groups may also be used ; these are those conventionally used in sugar
chemistry, for example according to G.J . Boons, Tetrahedron ( 1996), 52, 1095-1 121.
These activating groups are chosen , for example, from trichloroacetimidate groups and
thioglycosides.
The process described above makes it possible to obtain the compounds of the
invention in the form of salts, advantageously in the form of the sodium salt. To obtain the
corresponding acids, the compounds of the invention in salt form may be brought into
contact with a cation-exchange resin in acidic form. The compounds of the invention in
acid form may then be neutralized with a base so as to obtain the desired salt. For the
preparation of the salts of the compounds of formula ( I) , any inorganic or organic base
that gives pharmaceutically acceptable salts with the compounds of formula (I) may be
used .
A subject of the invention is also the compounds of formula ( II) below, in which Alk
represents an alkyl group and Pg and Pg' represent protecting groups as defined
previously:
In particular, a subject of the invention is the compound ( II) in which the Alk groups
represent methyl groups and Pg and Pg' represent, respectively, acetyl and
benzyloxycarbonyl groups (compound 17 in the synthesis schemes which follow).
A subject of the invention is also the compounds of formula ( III) below, in which Alk
represents an alkyl group, R is as previously defined in relation to the compounds of
formula (I), A represents an - NH-Pg" or -O-alkyl group, and Pg, Pg' and Pg" , which may
be identical to or different from one another, represent protecting groups as previously
defined :
In particular, a subject of the invention is the compounds (III) in which the Alk
groups represent methyl groups, R represents an -O-pentyl or -O-pentylphenyl group, Pg
represents an acetyl or benzoyl group, Pg' represents an acetyl or ie/f-butyldiphenylsilyl
group, and A represents an -NH-benzyloxycarbonyl or -O-butyl group.
More particularly, a subject of the invention is the compounds (III) in which:
- either Alk represents a methyl group, R represents an -O-pentyl group, Pg and
Pg' represent acetyl groups and A represents an -NH-benzyloxycarbonyl group
(compound 44 in the synthesis schemes which follow);
- or Alk represents a methyl group, R represents an -O-pentylphenyl group, Pg
and Pg' represent acetyl groups and A represents an -NH-benzyloxycarbonyl group
(compound 55 in the synthesis schemes which follow);
- or Alk represents a methyl group, R represents an -O-pentylphenyl group, Pg
represents an acetyl group, Pg' represents a ie/f-butyldiphenylsilyl group and A
represents an -NH-benzyloxycarbonyl group (compound 72 in the synthesis schemes
which follow);
- or Alk represents a methyl group, R represents an -O-pentylphenyl group, Pg
represents a benzoyl group, Pg' represents a ie/f-butyldiphenylsilyl group and A
represents an -O-butyl group (compound 97 in the synthesis schemes which follow).
A subject of the invention is also the compounds of formula (IV) below, in which
Alk represents an alkyl group, B represents an azide (N3) or -O-alkyI group, Pg, Pg' and
Pg", which may be identical to or different from one another, represent protecting groups
as previously defined, and D represents an activating group or an -O-acetyl group:
A subject of the invention is also the compounds of formula (IV) above, in which
Alk represents an alkyl group, B represents an azide (N3) or -O-alkyI group, Pg, Pg' and
Pg", which may be identical to or different from one another, represent protecting groups
as previously defined, and D represents an activating group or an -O-acetyl group, with
the exception of the compound of formula (IV) in which Alk represents a methyl group, B
represents an azide group, Pg represents a levulinyl group, Pg' and Pg" represent acetyl
groups, and D represents a trichloroacetimidate group.
Advantageously, the compounds of formula (IV) according to the invention are
such that B represents an -O-alkyI group.
Advantageously, the compounds of formula (IV) are such that the Alk groups
represent methyl groups, B represents an -O-butyl group, Pg represents a benzyl or
levulinyl group, Pg' represents an acetyl or benzoyl group, Pg" represents an acetyl or
fe/t-butyldiphenylsilyl group and D represents an activating group such as the
trichloroacetimidate (-0-C(NH)CCI 3) group or an -O-acetyl group.
In particular, a subject of the invention is the compounds (IV) in which the Alk
groups represent methyl groups, B represents an azide (N3) group, Pg represents a
benzyl or levulinyl group, Pg' represents an acetyl or benzoyl group, Pg" represents an
acetyl or fe/t-butyldiphenylsilyl group and D represents an activating group such as the
trichloroacetimidate (-0-C(NH)CCI 3) group or an -O-acetyl group, with the exception of
the compound of formula (IV) in which Alk represents a methyl group, B represents an
azide group, Pg represents a levulinyl group, Pg' and Pg" represent acetyl groups, and D
represents a trichloroacetimidate group.
More particularly, a subject of the invention is the compounds (IV) in which:
- either Alk represents a methyl group, B represents an azide (N3) group, Pg
represents a benzyl group, Pg' and Pg" represent acetyl groups, and D represents
an -0-C(NH)CCI 3 group (compound 28 in the synthesis schemes which follow);
- or Alk represents a methyl group, B represents an azide (N3) group, Pg
represents a levulinyl group, Pg' and Pg" represent acetyl groups, and D represents an
-0-C(NH)CCI 3 group (compound 29 in the synthesis schemes which follow);
- or Alk represents a methyl group, B represents an azide (N3) group, Pg
represents a levulinyl group, Pg' represents an acetyl group, Pg" represents a tertbutyldiphenylsilyl
group, and D represents an -0-C(NH)CCI 3 group (compound 69 in the
synthesis schemes which follow);
- or Alk represents a methyl group, B represents an -O-butyl group, Pg represents
a levulinyl group, Pg' represents a benzoyl group, Pg" represents an acetyl group, and D
represents an -O-acetyl group (compound 9 1 in the synthesis schemes which follow);
- or Alk represents a methyl group, B represents an -O-butyl group, Pg represents
a levulinyl group, Pg' represents a benzoyl group, Pg" represents an acetyl group, and D
represents an -0-C(NH)CCI 3 group (compound 93 in the synthesis schemes which
follow);
- or Alk represents a methyl group, B represents an -O-butyl group, Pg represents
a levulinyl group, Pg' represents a benzoyl group, Pg" represents a ie/f-butyldiphenylsilyl
group, and D represents an -0-C(NH)CCI 3 group (compound 101 in the synthesis
schemes which follow).
Such compounds of formulae (II), (III) and (IV) are of use as synthesis
intermediates for the compounds of formula (I).
The examples which follow describe the preparation of certain compounds in
accordance with the invention. These examples are not limiting, and merely illustrate the
present invention. The starting compounds and the reagents, when their mode of
preparation is not expressly described, are commercially available or described in the
literature, or else can be prepared according to methods which are described therein or
which are known to those skilled in the art.
The following abbreviations are used:
[a]D: optical rotation
Ac: acetyl
All: allyl
Bn: benzyl
BT: benzotriazole
Bz: benzoyl
TLC: Thin Layer Chromatography
DDQ: 2,3-dichloro-5,6-dicyano-1 ,4-benzoquinone
CE : Capillary Electrophoresis
ESI: Electron Spray Ionization
ESI-MS 2 : ESI coupled to Mass Spectrometry
Et: ethyl
h: hours
LC-MS : Liquid Chromatography coupled to Mass Spectrometry
Lev: levulinyl
Me: methyl
min: minute(s)
mL: millilitre(s)
mmol: millimol(s)
p: para
Phe: phenyl
Pent: pentyl
Rf: Retardation factor (retention time measured on TLC relative to the solvent
migration front)
NMR: Nuclear Magnetic Resonance
SFC: Supercritical Fluid Chromatography
SFC-MS 4 : SFC coupled to Mass Spectrometry
tert: tertiary
TBDMS: iert-butyldimethylsilyl
TBDPS: ie/f-butyldiphenylsilyl
THP: tetrahyd ropyran
Z: benzyloxycarbonyl
The capillary electrophoresis operations are carried out using a Beckman
apparatus under the following conditions: capillary: PVA Coated 40 cm (Ldet) c 50 m h (id),
electrolyte: 4 mM 5-sulphosalicylic acid, pH 3.51 (NaOH), detection: 214 nm indirect,
voltage: -15 kV, T° = 30°C, injection: 5 sec (0.5 psi), solution 0.5 mg/ml, coinjection: 5 sec
(0.5 psi) DMSO.
2 The ESI-MS spectra are recorded using an LCT apparatus (Waters) with a TOF
(Time-Of-Flight) analyser. The introduction mode is direct by infusion, the ionization mode
is by positive-mode or negative-mode electrospray as appropriate.
'The LC-MS are performed on a Waters ZQ4000 apparatus. The column used is
a Symetry C18 3.5 mhi (2.1 c 50 mm) column. Eluent A is made up of H20 + 0.005% TFA,
pH 3.15. Eluent B is made of acetonitrile + 0.005% TFA. The gradient ranges from 0 to
90% of eluent B over 10 (or 30) min + 5 min at 90% of eluent B. The flow rate is
0.4 ml/min.
4 The SFC-MS are carried out with a Mettler Toledo apparatus using a Diol 60A
5 m column (250 c 4.6 mm) - T° = 34°C - gas: C0 2 - modifier: 50% MeOH/50%
CH3CN - flow rate: 3 ml/min - pressure: 180 bar - gradient: 5% (2 min), 3%/min 35%
( 1 min) 95%/min 5%. Run time: 16 min. Mass spectrometry: positive electrospray.
Preparation of the synthesis intermediates :
SCHEME 1: Preparation of compound 17
Methyl 6-0-benzoyl-2-[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-a -Dglucopyranoside
(1 1
Triethylamine (8.3 ml, 59.9 mmol) and then BzOBt (13.6 g, 56.7 mmol) are added,
at ambient temperature, to a solution of methyl 2-[(benzyloxy)carbonyl]amino-2-deoxy-3-
O-methyl-a -D-glucopyranoside (10) (5.38 g, 15.8 mmol; described by Akiya, Shichiro and
Osawa, Toshiaki in Yakugaku Zasshi, 1956, 76, 1276-9) in dichloromethane (240 ml).
After stirring at ambient temperature for 16 hours, the mixture is diluted with
dichloromethane (800 ml). The organic phase is washed with a 2% aqueous solution of
sodium hydrogen carbonate and then with water, dried over sodium sulphate, filtered, and
then concentrated to dryness. Purification of the residue by flash chromatography on a
silica gel column (3/2 v/v dichloromethane/ethyl acetate) gives 5.65 g of compound 1 1 .
Rf = 0.42, silica gel, 5/1 v/v dichloromethane/ethyl acetate
Methyl (2-0-benzoyl-4,6-0-isopropylidene-3-0-methyl-a -L-idopyranosyl)-(1 ®-4)-6-0-
benzoyl-2-r(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-g -D-qlucopyranoside (13)
A mixture of ethyl 2-0-benzoyl-4,6-0-isopropylidene-3-0-methyl-1-thio-a-Lidopyranoside
12 (3.61 g, 9.44 mmol; prepared according to Jaurand, G. et al. Bioorg.
Med. Chem. Lett. 1992, 2, 897-900), of compound 11 (3.50 g, 7.86 mmol) and of 4A
molecular sieve powder ( 1 .90 g) in dichloromethane (62 ml) is stirred under an argon
atmosphere for 1 h. The mixture is then cooled to 0°C and /V-bromosuccinimide (4.03 g,
22.7 mmol) and trifluoromethanesulphonic acid (182 m I_, 2.08 mmol) are successively
added. After magnetic stirring for 45 min, sodium hydrogen carbonate is added and the
reaction mixture is then filtered and diluted with dichloromethane (450 ml). The organic
phase is washed with a 1M aqueous solution of sodium thiosulphate and then with water,
dried over sodium sulphate, filtered, and then concentrated to dryness. Purification of the
residue by flash chromatography on a silica gel column (1/1 v/v cyclohexane/ethyl
acetate) gives 5.66 g of compound 13.
Rf = 0.5, silica gel, 7/5 v/v cyclohexane/ethyl acetate
Methyl (4,6-0-isopropylidene-3-0-methyl-a -L-idopyranosyl)-(1 4)-2-
[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-a -D-glucopyranoside (14)
Potassium fe/t-butoxide (829 mg, 7.38 mmol) is added to a solution of compound
13 (5.65 g, 7.38 mmol) in a methanol-dioxane mixture (74 ml, 1/1 , v/v). The reaction
mixture is then stirred for 2 h at ambient temperature and then neutralized with Dowex
AG50WX4 resin, filtered, and then concentrated to dryness. The residue obtained is
purified by flash chromatography on a silica gel column (2/3 v/v dichloromethane/acetone),
to give 3.63 g of compound 14.
Rf = 0.56, silica gel, 2/1 v/v dichloromethane/acetone
Methyl (2-0-acetyl-4,6-0-isopropylidene-3-0-methyl-a -L-idopyranosyl)-(1 ®-4)-6-0-acetyl-
2-[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-a -D-glucopyranoside (15)
Compound 14 (3.62 g, 6.51 mmol) is dissolved in dichloromethane (26 ml), and
then triethylamine (2.7 ml, 19.5 mmol), 4-dimethylaminopyridine (80 mg, 0.65 mmol) and
acetic anhydride ( 1.8 ml, 18.2 mmol) are added. After stirring at 0°C for 10 minutes, and
then at ambient temperature for 2 h, the reaction mixture is diluted with dichloromethane
(500 ml) and then successively washed with a 10% aqueous solution of potassium
hydrogen sulphate, with water and with a 2% aqueous solution of sodium hydrogen
carbonate, and the organic phase is then dried over sodium sulphate, filtered and
concentrated. The resulting residue is purified by flash chromatography on a silica gel
column (3/7 v/v cyclohexane/ethyl acetate), to give 4.19 g of compound 15.
Rf = 0.48, silica gel, 5/7 v/v cyclohexane/ethyl acetate
Methyl (2-0-acetyl-3-0-methyl-a -L-idopyranosyl)-(1 ®-4)-6-0-acetyl-2-
(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-g -D-qlucopyranoside (16)
Compound 15 (4.18 g, 6.52 mmol) is dissolved in acetic acid (65 ml). The reaction
medium is stirred at ambient temperature for 16 h. After concentration under vacuum and
codistillation with toluene (4 c 100 ml), the resulting residue is purified by flash
chromatography on a silica gel column (1/4 v/v cyclohexane/acetone), to give 3.62 g of
compound 16.
Rf = 0.47, silica gel, 2/3 v/v cyclohexane/acetone
Methyl (methyl 2-0-acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 4)-6-0-acetyl-2-
[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-a -D-glucopyranoside (17)
A saturated aqueous solution of sodium hydrogen carbonate (24 ml) is added to a
solution of compound 16 (3.62 g, 6.02 mmol) in tetrahydrofuran (80 ml), and then, at 0°C
and under argon, a 0.32 M solution of 2,2,6, 6-tetramethylpiperidin-1-oxy (376 m I,
0.12 mmol) and a solution of 1,3-dibromo-5,5-dimethylhydantoin (10.3 ml, 12 mmol) are
successively added. After stirring at ambient temperature for 4 h 30, the reaction medium
is concentrated and then coevaporated with L/,/V-dimethylformamide (4 c 50 ml). The
residue obtained is placed in solution in L/,/V-dimethylformamide (80 ml) and potassium
hydrogen carbonate (3.01 g, 30.1 mmol) and then methyl iodide (3.7 ml, 60.2 mmol) are
added at 0°C and under argon. After completion of the reaction (TLC), the reaction
medium is concentrated under vacuum, and the reaction crude is diluted with ethyl
acetate (800 ml), washed with water and then with a 1M aqueous solution of sodium
thiosulphate, dried over sodium sulphate, filtered and concentrated. The resulting residue
is purified by chromatography on a Sephadex® LH20 column (190 c 3.2 cm, 1/1
dichloromethane/ethanol) followed by flash chromatography on a silica gel column (2/3 v/v
cyclohexane/acetone) to give 3.90 g of compound 17.
Rf = 0.36, silica gel, 1/1 v/v cyclohexane/acetone
SCHEME 2: Preparation of compound 28
28 27
(4,6-0-lsopropylidene-2-0-p-methoxybenzyl-3-0-methyl-a -L-idopyranosyl)-(1 ®-4)-1 ,6-
anhvdro-2-azido-2-deoxy-3-0-methyl-B -D-glucopyranose (1 9 )
p-methoxybenzyl chloride ( 1 .3 ml, 9.48 mmol) and then 55% sodium hydride
(370 mg, 7.70 mmol) are added, at 0°C and under argon, to a solution of compound 18
(2.47 g, 5.92 mmol; WO20 10/029 185) in L/,/V-dimethylformamide (24 ml). After stirring for
16 h, methanol is added, the reaction medium is concentrated under vacuum, and the
residue is diluted with ethyl acetate (500 ml), washed with water, dried over sodium
sulphate, filtered and concentrated. The residue obtained is purified by flash
chromatography on silica gel (55/45 v/v cyclohexane/acetone), to give 3.18 g of
compound 19.
Rf = 0.42, silica gel, 3/2 v/v cyclohexane/acetone
(2-0-p-Methoxybenzyl-3-0-methyl-a -L-idopyranosyl)-(1 ^4)-1 ,6-anhvdro-2-azido-2-deoxy-
3-0-methyl-p -D-qlucopyranose (20)
Compound 19 (3. 17 g, 5.91 mmol) is dissolved in acetic acid (60 ml). The reaction
medium is stirred at ambient temperature for 16 h. After concentration under vacuum and
codistillation with toluene (4 c 100 ml), the residue obtained is purified by flash
chromatography on a silica gel column (3/7 v/v cyclohexane/acetone), to give 2.66 g of
compound 20.
Rf = 0.45, silica gel, 1/ 1 v/v cyclohexane/acetone
(6-0-te/t-Butyldimethylsilyl-2-0-p-methoxybenzyl-3-0-methyl-a -L-idopyranosyl)-(1 ^4)-
1,6-anhvdro-2-azido-2-deoxy-3-0-methyl-p -D-qlucopyranose (21)
Compound 2 0 (2.67 g, 5.34 mmol) is dissolved in dichloromethane (53 ml), and
then triethylamine ( 1 .6 ml, 11.7 mmol), 4-dimethylaminopyridine (65 mg, 0.53 mmol) and
fe/t-butyldimethylsilyl chloride (886 mg, 5.87 mmol) are added . After stirring at 0°C for
30 minutes, and then at ambient temperature for 5 h, the same amount of reactants is
added. After stirring at ambient temperature for 16 h, the reaction mixture is diluted with
dichloromethane (500 ml), and then successively washed with a 10% aqueous solution of
potassium hydrogen sulphate and with water and then the organic phase is dried over
sodium sulphate, filtered and concentrated . The resulting residue is purified by flash
chromatography on a silica gel column (7/3 v/v cyclohexane/acetone), to give 3.46 g of
compound 21.
Rf = 0.50, silica gel, 2/1 v/v cyclohexane/acetone
(4-0-Benzyl-6-0-te/t-butyldimethylsilyl-2-0-p-methoxybenzyl-3-0-methyl-a -LidopyranosvQ-
d 4)-1 ,6-anhvdro-2-azido-2-deoxy-3-0-methyl-B -D-glucopyranose (22)
Benzyl bromide ( 1 .3 ml, 26.7 mmol) and then 55% sodium hydride (385 mg,
8.01 mmol) are added, at 0°C and under argon, to a solution of compound 2 1 (3.26 g,
5.34 mmol) in L/,/V-dimethylformamide (27 ml). After stirring for 3 h, methanol (3 ml) is
added, the reaction medium is concentrated under vacuum, and the residue is diluted with
ethyl acetate (500 ml), washed with water, dried over sodium sulphate, filtered and
concentrated under vacuum. The residue then obtained is purified by flash
chromatography on a silica gel column (7/3 v/v cyclohexane/acetone), to give 3.67 g of
compound 22.
Rf = 0.54, silica gel, 5/2 v/v cyclohexane/acetone
(4-0-Benzyl-6-0-te/t-butyldimethylsilyl-3-0-methyl-a -L-idopyranosyl)-(1^4)-1 ,6-anhvdro-
2-azido-2-deoxy-3-0-methyl-p -D-qlucopyranose (23)
Water ( 10 ml) and then, at 0°C, DDQ ( 1 .78 g, 7.85 mmol) are added to a solution
of compound 22 (3.67 g, 5.23 mmol) in dichloromethane (210 ml). After stirring at 0°C for
5 h 30, the medium is diluted with dichloromethane (700 ml) and a 2% aqueous solution of
sodium hydrogen carbonate is added. The organic phase is then washed with water, dried
over sodium sulphate, filtered and concentrated. The residue obtained is purified by flash
chromatography on a silica gel column (7/3 v/v toluene/ethyl acetate), to give 2.87 g of
compound 23.
Rf = 0.45, silica gel, 2/1 v/v toluene/acetone
(2-0-Acetyl-4-0-benzyl-6-0-te/t-butyldimethylsilyl-3-0-methyl-a -L-idopyranosyl)-(1 ®-4)-
1,6-anhvdro-2-azido-2-deoxy-3-0-methyl-p -D-qlucopyranose (24)
Compound 23 (2.86 g, 4.92 mmol) is dissolved in dichloromethane (20 ml), and
then triethylamine ( 1.0 ml, 7.37 mmol), 4-dimethylaminopyridine (60 mg, 0.50 mmol) and
acetic anhydride (650 m I, 6.88 mmol) are added. After stirring at 0°C for 1 h and then at
ambient temperature for 16 h, the reaction mixture is diluted with dichloromethane (50 ml),
and then successively washed with a 10% aqueous solution of potassium hydrogen
sulphate and with water, and then the organic phase is dried over sodium sulphate,
filtered and concentrated. The resulting residue is purified by flash chromatography on a
silica gel column (7/3 v/v toluene/ethyl acetate), to give 3.46 g of compound 24.
Rf = 0.6, silica gel, 2/1 v/v toluene/ethyl acetate
(Methyl 2-0-acetyl-4-0-benzyl-3-0-methyl-a -L-idopyranosyluronate)-(1 4)-1,6-anhydro-
2-azido-2-deoxy-3-0-methyl-B -D-glucopyranose (25)
A solution of chromium trioxide ( 1 .2 g) in 3.5M sulphuric acid (5.4 ml) is added, at
0°C, to a solution of compound 24 (3.0 g, 4.83 mmol) in acetone (193 ml). After stirring at
0°C for 5 h 30, the reaction medium is diluted with dichloromethane (800 ml), washed with
water, dried over sodium sulphate, filtered and concentrated. The compound obtained is
used in the next step without purification. The residue obtained is dissolved in N,Ndimethylformamide
(63 ml), and potassium hydrogen carbonate (2.42 g, 24.1 mmol) and
also methyl iodide (3.0 ml, 48.3 mmol) are added at 0°C. The reaction mixture is stirred at
ambient temperature for 4 h, and then concentrated under vacuum. The residue is diluted
with ethyl acetate (800 ml) and then washed with water, with a saturated aqueous solution
of sodium thiosulphate and with a saturated aqueous solution of sodium chloride, and
then dried over sodium sulphate, filtered and concentrated. The resulting residue is
purified by flash chromatography on a silica gel column (3/2 v/v toluene/ethyl acetate), to
give 2.08 g of compound 25.
Rf = 0.47, silica gel, 1/1 v/v toluene/ethyl acetate
(Methyl 2-0-acetyl-4-0-benzyl-3-0-methyl-a -L-idopyranosyluronate)-(1 4)-1,6-di-Oacetyl-
2-azido-2-deoxy-3-0-methyl-g,p -D-qlucopyranose (26)
Trifluoroacetic acid (923 m I, 12 mmol) is added, at 0°C, to a solution of compound
25 (585 mg, 1.09 mmol) in acetic anhydride (10.3 ml). The reaction medium is stirred for
4 h at ambient temperature. After concentration under vacuum, the mixture is
coevaporated with toluene. Purification of the residue by chromatography on a silica gel
column (toluene/acetone) gives 694.5 mg of compound 26.
H NMR [500 MHz] (CDCI3) d of the anomeric protons:
Glc ' b 5.42 ppm, Glc ' a 6.17 ppm and IdoUA" 5.1 ppm.
(Methyl 2-0-acetyl-4-0-benzyl-3-0-methyl-a -L-idopyranosyluronate)-(1^4)-6-0-acetyl-2-
azido-2-deoxy-3-0-methyl-a,B -D-glucopyranose (27)
Benzylamine (4.5 ml, 4 1.2 mmol) is added, under an argon atmosphere at 0°C, to
a solution of compound 26 (694 mg, 1.09 mmol) in diethyl ether (32 ml). The reaction
medium is stirred for 3 h at ambient temperature and then stored at +4°C for 2 1 h. After
dilution with ethyl acetate, the reaction medium is successively washed with an aqueous
solution of hydrochloric acid ( 1M) and then with water. The organic phase is dried over
sodium sulphate, filtered and concentrated under vacuum. The resulting residue is purified
by chromatography on a silica gel column, to give 578.1 mg of compound 27.
H NMR [500MHz] (CDCI3) d of the anomeric protons:
Glc ' b 4.56 ppm, Glc ' a 5.26 ppm and IdoUA" 5.12 ppm.
(Methyl 2-0-acetyl-4-0-benzyl-3-0-methyl-a -L-idopyranosyluronate)-(1^4)-6-0-acetyl-2-
azido-2-deoxy-3-0-methyl-a,B -D-glucopyranose trichloroacetimidate (28)
Trichloroacetonitrile (476 m I, 4.74 mmol) and caesium carbonate (469 mg,
1.44 mmol) are added at 0°C to a solution of compound 27 (566.9 mg, 0.95 mmol) in
dichloromethane (19 ml) in the presence of 4 A molecular sieve powder (950 mg). After
stirring at ambient temperature for 16 h, the reaction medium is filtered through Celite®
and then concentrated. The residue is purified by chromatography on a silica gel column,
to give 608 mg of compound 28.
H NMR [500 MHz] (CDCI3) d of the anomeric protons:
Glc ' b 5.60 ppm, Glc ' a 6.30 ppm and IdoUA" 5.13 ppm.
SCHEME 3: Preparation of the octasaccharide 34
Methyl (methyl 2-0-acetyl-4-0-levulinoyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ®4 )-(6-
0-acetyl-2-azido-2-deoxy-3-0-methyl-a -D-glucopyranosyl)-(1^4)-(methyl 2-Q-acetyl-3-0-
methyl-a -L-idopyranosyluronate)-(1^4)-6-0-acetyl-2-[(benzyloxy)carbonyllamino-2-
deoxy-3-O-methyl-a -D-glucopyranoside (30)
A mixture of compound 29 (363 mg, 0.49 mmol) (described in WO201 0/0291 85),
of the glycosyl acceptor 17 (489 mg, 0.77 mmol) and of 4 A molecular sieve powder
(363 mg) in dichloromethane (40 ml) is stirred under an argon atmosphere for 1 h at 25°C.
The reaction mixture is cooled to -25°C and a 1M solution of ie/f-butyldimethylsilyl triflate
in dichloromethane (73 m I) is added to the reaction medium. After stirring for 15 minutes,
the reaction medium is neutralized by adding solid sodium hydrogen carbonate. After
filtration and concentration, the organic phase is washed with a 2% aqueous solution of
sodium hydrogen carbonate and with water, dried over sodium sulphate, filtered and then
concentrated to dryness. The residue obtained is purified by size exclusion
chromatography (Sephadex® LH20, 190 c 3.2 cm, 1/1 v/v dichloromethane/ethanol), to
give 393 mg of compound 30.
H NMR [500 MHz] (CDCI3) d of the anomeric protons
Glc ' a 4.64 ppm, IdoUA" a 5.07 ppm, Glc '" a 4.97 ppm and ldoUAlv a 5.05 ppm.
Methyl (methyl 2-0-acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ^4)-(6-0-acetyl-2-
azido-2-deoxy-3-0-methyl-a -D-qlucopyranosyl)-(1^4)-(methyl 2-0-acetyl-3-0-methyl-a -Lidopyranosyluronate)-(
1 ®-4)-6-0-acetyl-2-r(benzyloxy)carbonyllamino-2-deoxy-3-0-
methyl-g -D-qlucopyranoside (31)
Hydrazine acetate (253 mg, 2.75 mmol) is added to a solution of compound 30
(670 mg, 0.55 mmol) in a 1/2 v/v toluene/ethanol mixture (290 ml). The reaction medium is
stirred for 1 h at ambient temperature. After concentration, the residue is purified by flash
chromatography on a silica gel column (1/9 v/v toluene/ethyl acetate), to give 677 mg of
compound 31.
H NMR [500 MHz] (CDCI3) d of the anomeric protons
Glc ' a 4.64 ppm, IdoUA" a 5.08 ppm, Glc '" a 4.98 ppm and ldoUAlv a 4.98 ppm.
Methyl (methyl 2-0-acetyl-4-0-levulinoyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ^4)-[(6-
0-acetyl-2-azido-2-deoxy-3-0-methyl-a -D-qlucopyranosyl)-(1^4)-(methyl 2-Q-acetyl-3-0-
methyl-a -L-idopyranosyluronate)-(1^4)1?-6-0-acetyl-2-[(benzyloxy)carbonyl1amino-2-
deoxy-3-O-methyl-a -D-qlucopyranoside (32)
A mixture of compound 29 (442 mg, 0.59 mmol), of the glycosyl acceptor 3 1
(677 mg, 0.60 mmol) and of 4 A molecular sieve powder (442 mg) in dichloromethane
(21 ml) is stirred under an argon atmosphere for 1 h at 25°C. The reaction mixture is
cooled to -25°C and a 1M solution of ie/f-butyldimethylsilyl triflate in dichloromethane
(90 m I) is added to the reaction medium. After stirring for 15 minutes, the reaction medium
is neutralized by adding solid sodium hydrogen carbonate. After filtration and
concentration, the organic phase is washed with a 2% aqueous solution of sodium
hydrogen carbonate and with water, dried over sodium sulphate, filtered and then
concentrated to dryness. The residue obtained is purified by size exclusion
chromatography (Sephadex® LH20, 190 c 3.2 cm, 1/1 v/v dichloromethane/ethanol), to
give 564 mg of compound 32.
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc ' a
4.64 ppm, IdoUA" a 5.08 ppm, Glc '" a 5.0 ppm, ldoUA lv a 5.08 ppm,
Glc v a 4.98 ppm and ldoUAv l a 5.07 ppm.
Methyl (methyl 2-0-acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 4)-[(6-Q-acetyl-2-
azido-2-deoxy-3-0-methyl-a -D-qlucopyranosyl)-(1 4)-(methyl 2-0-acetyl-3-0-methyl-a -Lidopyranosyluronate)-(
1 ®-4 )l -6-0-acetyl-2-[(benzyloxy)carbonyllamino-2-deoxy-3-0-
methyl-a -D-glucopyranoside (33)
Hydrazine acetate (152 mg, 1.65 mmol) is added to a solution of compound 32
(564 mg, 0.33 mmol) in a 1/2 v/v toluene/ethanol mixture (66 ml). The reaction medium is
stirred at ambient temperature for 1 h. After concentration, the residue is purified by flash
chromatography on a silica gel column (1/9 v/v toluene/ethyl acetate), to give 480 mg of
compound 33.
Rf = 0.49, silica gel, 1/9 v/v toluene/ethyl acetate.
Methyl (methyl 2-0-acetyl-4-0-benzyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ^4)- G(6-0-
acetyl-2-azido-2-deoxy-3-0-methyl-a -D-qlucopyranosyl)-(1^4)-(methyl 2-Q-acetyl-3-0-
methyl-a -L-idopyranosyluronate)-(1 ®-4)ly6-0-acetyl-2 - (benzyloxy)carbonyllamino-2-
deoxy-3-O-methyl-a -D-qlucopyranoside (34)
A mixture of compound 28 (332 mg, 0.447 mmol), of the glycosyl acceptor 33
(480 mg, 0.298 mmol) and of 4 A molecular sieve powder (224 mg) in dichloromethane
( 1 1 ml) is stirred under an argon atmosphere for 1 h at ambient temperature. The reaction
mixture is cooled to -20°C and a 0.1 M solution of ie/f-butyldimethylsilyl triflate in
dichloromethane (4.5 ml) is added to the reaction medium. After 1 h 30 min, the reaction
medium is neutralized by adding solid sodium hydrogen carbonate. After filtration and
concentration under vacuum, the organic phase is washed with a 2% aqueous solution of
sodium hydrogen carbonate and with water, dried over sodium sulphate, filtered and then
concentrated to dryness. The residue obtained is purified by chromatography on a silica
gel column, to give 500 mg of compound 34.
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc ' a
4.65 ppm, IdoUA" a 5.10 ppm, Glc '" a 4.97 ppm, ldoUA lv a 5.10 ppm,
Glc v a 4.98 ppm, ldoUAv l a 5.1 0 ppm, Glc v"a 5.00 ppm and
ldoUAv l" 5.12 ppm.
SCHEME 4: Preparation of the octasaccharide 38
Methyl (methyl 4-0-benzyl-3-0-methyl-a -L-idopyranosyluronate)-(1 4)-[(2-azido-2-deoxy-
3-0-methyl-a -D-glucopyranosyl)-(1 4)-(methyl 3-O-methyl-a -L-idopyranosyluronate)-
( 1®-4)lg-2-[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-a -D-glucopyranoside (35)
A 1M solution of sodium methoxide in methanol (684 m I) is added, at 0°C, under an
argon atmosphere, to a solution of compound 34 (500 mg, 0.228 mmol) in a 2/3 v/v
dichloromethane/methanol mixture (68 ml) containing 3A molecular sieve (285 mg). After
magnetic stirring at ambient temperature for 18 h, the reaction medium is neutralized with
Dowex® 50WX4 H+ resin. After filtration and concentration under vacuum, the residue is
purified by chromatography on a silica gel column, to give 170 mg of compound 35.
Rf = 0.54, silica gel, 9/1 v/v dichloromethane/methanol.
Methyl (methyl 4-0-benzyl-3-0-methyl-2-0-triethylammonium sulphonato-g -Lidopyranosyluronate)-(
1^4)-[(2-azido-2-deoxy-3-0-methyl-6-0-triethylammonium
sulphonato-a -D-glucopyranosyl)-(1 4)-(methyl 3-0-methyl-2-0-triethylammonium
sulphonato-a -L-idopyranosyluronate)-(1 ®-4 ) l -2-[(benzyloxy)carbonyllamino-2-deoxy-3-0-
methyl-6-O-triethylammonium sulphonato-g -D-glucopyranoside (36)
Compound 35 (84.5 mg, 0.046 mmol) is dried by codistillation of anhydrous N,Ndimethylformamide
(3 x 4 ml) and is then placed in solution in anhydrous N,Ndimethylformamide
(4 ml). The sulphur trioxide-trithylamine complex (331 mg,
1.825 mmol) is added to this solution. The mixture is stirred for 16 h at 55°C in the dark
and then the excess reagent is destroyed with methanol (224 m I, 5.52 mmol). The reaction
medium is loaded onto a Sephadex ® LH20 gel column (95 c 2 cm) eluted with a
75/20/5 v/v/v methanol//V,/V-dimethylformamide/H 20 mixture, to give compound 36
(142 mg).
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc ' a
4.59 ppm, IdoUA" a 5.31 ppm, Glc '" a 5.21 ppm, ldoUA lv a 5.32 ppm,
Glc v a 5.20 ppm, ldoUA v l a 5.32 ppm, Glc v"a 5.1 7 ppm and
ldoUAv l" a 5.34 ppm.
Methyl (lithium 4-0-benzyl-3-0-methyl-2-0-lithium sulphonato-a -L-idopyranosyluronate)-
( 1 4H(2-azido-2-deoxy-3-0-methyl-6-0-lithium sulphonato-a -D-qlucopyranosyl)-(1 ®4)-
(rnethyl lithium 3-Q-methyl-2-0-lithium sulphonato-a -L-idopyranosyluronate)-(1 4 ΐ -2-
[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-6-0-lithium sulphonato-g -Dglucopyranoside
(37)
A 0.5 M solution of lithium hydroxide in water (4.1 ml, 2.075 mmol) is added, at
0°C under argon, to compound 36 (38 mg, 0.0152 mmol) dissolved in a 1: 1
methanol/tetrahydrofuran solution (8.3 ml). After stirring at 0°C for 19 h, the reaction
medium is loaded onto a column of Sephadex ® LH20 gel (95 c 2 cm) eluted with a 75/20/5
v/v/v methanol//V,/V-dimethylformamide/water mixture, to give compound 37 (161 mg).
Rf = 0.07, silica gel, ethyl acetate/pyridine/acetic acid/water (6/2/2/0.6/1 )/(5/5/1/3)
9/1 v/v.
Methyl (sodium 3-0-methyl-2-0-sodium sulphonato-a -L-idopyranosyluronate)-(1 4)-[(2-
amino-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a -D-glucopyranosyl)-(1 4)-(methyl
sodium 3-Q-methyl-2-0-sodium sulphonato-a -L-idopyranosyluronate)-(1 4)l:r2-amino-2-
deoxy-3-0-methyl-6-0-sodium sulphonato-g -D-glucopyranoside (38)
Ammonium formate (194 mg, 3.071 mmol) and 10% palladium-on-carbon
(385 mg) are added, under an inert atmosphere, to a solution of compound 37 ( 154 mg,
0.061 mmol) in 1/1 v/v fe/t-butanol/water (12 ml). After stirring at ambient temperature for
4 h 15 min, the reaction medium is filtered (Millipore ® filter LSWP 5 m h ) and concentrated
to dryness. The residue is loaded onto a column of Sephadex ® G25-fine gel (95 c 2 cm)
eluted with a 0.2 M aqueous solution of NaCI. The fractions containing the expected
compound are combined, and loaded onto a column of Sephadex ® G25-fine gel
(95 x 2 cm) eluted with water. The product 38 is obtained (85.5 mg).
H NMR [500 MHz] (D20 ) d of the anomeric protons Glc ' a
4.97 ppm, IdoUA" a 5.32 ppm, Glc '" a 5.14 ppm, ldoUA lv a 5.32 ppm,
Glc v a 5.14 ppm, ldoUA v l 5.32 ppm, Glc v " a 5.14 ppm and
ldoUAv l" a 5.21 ppm.
SCHEME 5: Preparation of the disaccharide 44
Methyl (methyl 2-0-acetyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1 ®4 -6-
0-acetyl-2-r(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-g-D-qlucopyranoside (39)
4-dimethylaminopyridine (76 mg, 0.626 mmol), 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide hydrochloride ( 1.2 g, 6.26 mmol) and levulinic acid (643 m I, 6,26 mmol)
are added successively, under an inert atmosphere, to a solution of compound 17 in
dioxane (63 ml). After stirring at ambient temperature for 5 h 45 min,
4-dimethylaminopyridine (38 mg, 0.313 mmol), 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide hydrochloride (0.6 g, 3.13 mmol) and levulinic acid (322 m I, 3.13 mmol)
are again successively added. After stirring at ambient temperature for 16 h, the reaction
medium is concentrated and the residue is placed in solution in dichloromethane. The
organic phase is washed successively with a 10% aqueous solution of potassium
hydrogen sulphate, with a 2% aqueous solution of sodium hydrogen carbonate and then
with a saturated solution of sodium chloride. The organic phase is then dried over sodium
sulphate, filtered and then evaporated to dryness. The residue is purified by flash
chromatography on a silica gel column (cyclohexane/acetone), to give 2.23 g of
compound 39.
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc ' 4.96 ppm and IdoUA"
5.07 ppm.
(Methyl 2-0-acetyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1 4)-1 ,6-di-Oacetyl-
2-[(benzyloxy)carbonyllarriino-2-deoxy-3-0-methyl-a,B-D-glucopyranose (40)
96% sulphuric acid (179 m I) previously diluted in a 1/1 acetic acid/acetic anhydride
solution ( 1 .8 ml) is added, at 0°C under an argon atmosphere, to a solution of compound
39 ( 1.29 g, 1.79 mmol) in a 1/1 acetic acid/acetic anhydride solution (27 ml). After stirring
at ambient temperature for 3 h 30 min, the progression of the reaction is stopped by
adding triethylamine (25 ml). The reaction medium is coevaporated with toluene. The
residue obtained is purified by flash chromatography on a silica gel column
(dichloromethane/acetone), to give 1.59 g of compound 40.
40a LC-MS m/z 736.2 [(M + Na)+] . TR = 8.123 min
40b LC-MS m/z 736.2 [(M + Na)+] . TR = 8.043 min
(Methyl 2-0-acetyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1^4)-6-0-acetyl-
2- (benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-g.p-D-qlucopyranose (41)
Benzylamine (2.7 ml, 25.1 mmol) and acetic acid (38 m I, 0.662 mmol) are added
successively, under an argon atmosphere, to a solution of compound 40 (500 mg,
0.662 mmol) in tetrahydrofuran (26.5 ml). After magnetic stirring for 9 h, the reaction
medium is neutralized with Dowex AG 50 WX4 H+ resin, filtered and then concentrated.
The residue is purified by flash chromatography on a silica gel column (toluene/acetone),
to give 335 mg of compound 4 1.
Rf = 0.3, silica gel, 1/1 v/v toluene/acetone
(Methyl 2-0-acetyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1^4)-6-0-acetyl-
2- (benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-g.p-D-qlucopyranose
trichloroacetimidate (42)
Trichloroacetonitrile (528 m I, 5.27 mmol) and caesium carbonate (233 mg,
1.69 mmol) are added, at ambient temperature under an argon atmosphere, to a solution
of compound 4 1 (752.5 mg, 1.05 mmol) in dichloromethane (21 ml). After stirring at
ambient temperature for 16 h, the reaction medium is filtered through Celite® and then
concentrated. The residue is purified by chromatography on a silica gel column
(toluene/acetone + 0.1% triethylamine), to give 633 mg of compound 42.
42a H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc': 6.27 ppm and
ldoUA" 5.07 ppm.
42b H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc': 5.95 ppm and
ldoUA" 5.07 ppm.
Pent-4-ene (methyl 2-0-acetyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-
(1^4)-6-0-acetyl-2-[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-B-D-glucopyranoside
143}
A mixture of trichloroacetimidate 42 (130.9 mg, 0.153 mmol), of 4-penten-1-ol
(78 m I_, 0.763 mmol) and of 4 A molecular sieve powder ( 130 mg) in dichloromethane
(6.9 ml) is stirred under an argon atmosphere for 1 h 45 min at ambient temperature. The
reaction mixture is cooled to -20°C and a 1M solution of ie/f-butyldimethylsilyl triflate in
dichloromethane (30.5 m I_, 0.0305 mmol) is added dropwise. After stirring at -20°C for
35 min, a further addition of solution of ie/f-butyldimethylsilyl triflate in dichloromethane
( 15 m I, 0.01 5 mmol) is carried out. After stirring at -20°C for 10 min, the reaction is
neutralized by adding solid sodium hydrogen carbonate. The reaction medium is filtered
through Celite® and then evaporated. The residue is purified by chromatography on a
silica gel column (dichloromethane/acetone), to give 379 mg of compound 43.
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc ': 4.63 ppm and IdoUA"
5.04 ppm.
Pent-4-ene (methyl 2-0-acetyl-3-0-methyl-a-L-idopyranosyluronate)-(1 ^4)-6-Q-acetyl-2-
(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-p-D-qlucopyranoside (44)
Hydrazine acetate (220 mg, 2.4 mmol) is added to a solution of compound 43 in a
1/2 toluene/ethanol mixture (96 ml). The reaction medium is stirred for 40 min at ambient
temperature. After concentration, the residue is taken up in dichloromethane and then
washed with water. After drying over sodium sulphate, filtration and then concentration,
the residue is chromatographed on a silica gel column (dichloromethane/acetone), to give
315 mg of compound 44 .
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc ' 4.66 ppm and IdoUA"
4.99 ppm
SCHEME 6: Preparation of the octasaccharide 49
Pent-4-ene (methyl 2-0-acetyl-3-0-methyl-4-0-levulinoyl-a -L-idopyranosyluronate)-
( 1®-4)-(6-0-acetyl-2-azido-2-deoxy-3-0-methyl-a -D-glucopyranosyl)-(1 4)-(methyl 2-0-
acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ®-4)-6-0-acetyl-2-
[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-B -D-glucopyranoside (45)
Compound 29 (427 mg, 0.57 mmol) and compound 44 (300 mg, 0.439 mmol) are
processed according to the same procedure as that described for the preparation of 30, to
give compound 45 (340 mg).
H NMR [500 MHz] (CDCI3) d of the anomeric protons
Glc': 4.68, IdollA": 5.06, Glc : 4.98 and ldoUA lv : 5.08.
Pent-4-ene (methyl 2-0-acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 4)-(6-0-acetyl-2-
azido-2-deoxy-3-0-methyl-a -D-qlucopyranosyl)-(1 4)-(methyl 2-0-acetyl-3-0-methyl-a -Lidopyranosyluronate)-(
1 ®-4)-6-0-acetyl-2-[(benzyloxy)carbonyllamino-2-deoxy-3-0-
methyl-B-D-glucopyranoside (46)
Compound 45 (374 mg, 0.294 mmol) is processed according to the same
procedure as that described for the preparation of 3 1, to give compound 46 (386 mg).
H NMR [500 MHz] (CDCI3) d of the anomeric protons
Glc': 4.68, IdollA": 5.06, Glc'": 4.99 and ldoUA lv : 5.00.
Pent-4-ene (methyl 2-0-acetyl-4-0-levulinoyl-3-0-methyl-a -L-idopyranosyluronate)-
( 1®-4)-(6-0-acetyl-2-azido-2-deoxy-3-0-methyl-a -D-qlucopyranosyl)-(1 4)-(methyl 2-0-
acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ®-4)-(6-0-acetyl-2-azido-2-deoxy-3-0-
methyl-a -D-qlucopyranosyl)-(1 4)-(methyl 2-0-acetyl-3-0-methyl-a -Lidopyranosyluronate)-(
1 ®-4)-6-0-acetyl-2-[(benzyloxy)carbonyl1amino-2-deoxy-3-0-
meth l- -D-qlucopyranoside (47)
Compound 29 (75.5 mg, 0.101 mmol) and compound 46 (91 mg, 0.0776 mmol)
are processed according to the same procedure as that described for the preparation of
32, to give compound 47 (388 mg).
H NMR [500 MHz] (CDCI3) d of the anomeric protons
Glc': 4.67, IdoUA": 5.07, Glc'": 4.98, ldoUA lv : 5.10, Glcv : 5.02 and ldoUAv l: 5.08.
Pent-4-ene (methyl 2-0-acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 4)-(6-0-acetyl-2-
azido-2-deoxy-3-0-methyl-a -D-qlucopyranosyl)-(1 4)-(methyl 2-0-acetyl-3-0-methyl-a -Lidopyranosyluronate)-(
1 ®-4)-(6-0-acetyl-2-azido-2-deoxy-3-0-methyl-a -Dqlucopyranosyl)-(
1 4)-(methyl 2-0-acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ®-4)-6-
0-acetyl-2-[(benzyloxy)carbonyl1amino-2-deoxy-3-0-methyl-B -D-qlucopyranoside (48)
Compound 47 (382 mg, 0.217 mmol) is processed according to the same
procedure as that described for the preparation of 33, to give compound 48 (327 mg).
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc': 4.67 IdoUA": 5.07,
Glc'": 4.98, ldoUA lv : 5.09, Glcv : 5.02, ldoUAv l: 5.00.
Pent-4-ene (methyl 2-0-acetyl-4-0-benzyl-3-0-methyl-a -L-idopyranosyluronate)-(1 4)-
(6-0-acetyl-2-azido-2-deoxy-3-0-methyl-a -D-glucopyranosyl)-(1 4) (methyl 2-0-acetyl-3-
0-methyl-a -L-idopyranosyluronate)-(1^4)-(6-0-acetyl-2-azido-2-deoxy-3-0-methyl-a -Dglucopyranosyl)-(
1 4)-(methyl 2-0-acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ®-4)-
(6-0-acetyl-2-azido-2-deoxy-3-0-methyl-a -D-glucopyranosyl)-(1 ®-4)-(methyl 2-Q-acetyl-3-
0-methyl-a -L-idopyranosyluronate)-(1^4)-6-0-acetyl-2-[(benzyloxy)carbonyllamino-2-
deoxy-3-0-methyl-B -D-glucopyranoside (49)
Compound 28 (185.9 mg, 0.251 mmol) and compound 48 (321 mg, 0.193 mmol)
are processed according to the same procedure as that described for the preparation of
34, to give compound 49 (209 mg).
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc': 4.67 ppm, IdollA":
5.07 ppm, Glc"': 4.98 ppm, ldoUA lv : 5.09 ppm, Glcv : 5.02 ppm, ldoUAv l: 5.08 ppm,
Glcv " : 5.00 ppm and ldoUAv l" : 5.13 ppm.
SCHEME 7: Preparation of the octasaccharide 53
Pent-4-ene (methyl 4-0-benzyl-3-0-methyl-a -L-idopyranosyluronate)-(1 4)-(2-azido-2-
deoxy-3-0-methyl-a -D-glucopyranosyl)-(1 4)-(methyl 3-O-methyl-a -Lidopyranosyluronate)-(
1 ®-4)-(2-azido-2-deoxy-3-0-methyl-a -D-glucopyranosyl)-(1 ®-4)-
(methyl 3-0-methyl-a -L-idopyranosyluronate)-(1^4)-(2-azido-2-deoxy-3-0-methyl-a -DglucopyranosvQ-
d 4)-(methyl 3-0-methyl-a -L-idopyranosyluronate)-(1 4)-2-
[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-B -D-glucopyranoside (50)
A 0.5M solution of sodium methoxide in methanol (67 m I) is added, at 0°C, under
an argon atmosphere, to a solution of compound 49 (50 mg, 0.0223 mmol) in a 2/3 v/v
dichloromethane/methanol mixture (6.7 ml) containing 3A molecular sieve (29 mg). After
magnetic stirring at 0°C for 3 h, at ambient temperature for 4 h 45 min, at -18°C for 16 h,
and then at ambient temperature for 2 h, the reaction medium is neutralized with Dowex
50WX4 H+ resin. After filtration and concentration under vacuum, the residue is purified by
size exclusion chromatography (Sephadex ® LH20, 120 c 3 cm, 7/2/1 methanol//V,/Vdimethylformamide/
water), to give 38.7 mg of compound 50.
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc': 4.69, IdoUA": 5.16,
Glc'": 5.05, ldoUA lv : 5.18, Glcv : 5.06, ldoUAv l: 5.18, Glcv " : 5.04 and ldoUAv l" : 5.19.
Pent-4-ene (methyl 4-0-benzyl-3-0-methyl-2-0-triethylammonium sulphonato -g-Lidopyranosyluronate)-(
1^4)-(2-azido-2-desoxy-3-0-methyl-6-0-triethylammonium
sulphonato -g-D-glucopyranosyl)-(1 4)-(methyl 3-0-methyl-2-0-triethylammonium
sulphonato-a -L-idopyranosyluronate)-(1 ®-4)-(2-azido-2-desoxy-3-0-methyl-6-0-
triethylammonium sulphonato-a -D-glucopyranosyl)-(1 4)-(methyl 3-0-methyl-2-0-
triethylammonium sulphonato -g-L-idopyranosyluronate)-(1 4)-(2-azido-2-desoxy-3-0-
methyl-6-O-triethylammonium sulphonato -g-D-glucopyranosyl)-(1 4)-(methyl 3-O-methyl-
2-O-triethylammonium sulphonato-a -L-idopyranosyluronate)-(1^4)-2-
(benzyloxy)carbonyllamino-2-desoxy-3-0-methyl-6-0-triethylammonium sulphonato- b- -
qlucopyranoside (51)
Compound 50 (37.5 mg, 0.0197 mmol) is processed according to the same
procedure as that described for the preparation of 36, to give compound 5 1 (56.1 mg).
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc': 4.39, IdoUA": 5.23,
Glc'": 5.15, ldoUA lv : 5.30, Glcv : 5.15, ldoUAv l: 5.30, Glcv " : 5.13 and ldoUAv l" : 5.32.
Pent-4-ene (lithium 4-0-benzyl-3-0-methyl-2-0-lithium sulphonato -g-Lidopyranosyluronate)-(
1^4)-(2-azido-2-deoxy-3-0-methyl-6-0-lithium sulphonato -g-Dglucopyranosyl)-(
1 4)-(lithium 3-Q-methyl-2-0-lithium sulphonato -g-Lidopyranosyluronate)-(
1 4)-(2-azido-2-deoxy-3-0-methyl-6-0-lithium sulphonato -g-Dglucopyranosyl)-(
1 4)-(lithium 3-0-methyl-2-0-lithium sulphonato -g-Lidopyranosyluronate)-(
1^4)-(2-azido-2-deoxy-3-0-methyl-6-0-lithium sulphonato -g-Dglucopyranosyl)-(
1 4)-(lithium 3-Q-methyl-2-0-lithium sulphonato -g-Lidopyranosyluronate)-(
1^4)-2-[(benzyloxy)carbonyl1amino-2-deoxy-3-0-methyl-6-0-
lithium sulphonato -B-D-glucopyranoside (52)
Compound 5 1 (71 .3 mg, 0.0212 mmol) is processed according to the same
procedure as that described for the preparation of 37, to give compound 52 (58.7 mg).
Rf = 0.29, silica gel, ethyl acetate/pyridine/acetic acid/water (6/2/2/0.6/1 )/(5/5/1/3)
1/8 v/v.
Pentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a -L-idopyranosyluronate)-(1 4)-(2-
amino-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a -D-glucopyranosyl)-(1 4)-(sodium
3-Q-methyl-2-0-sodium sulphonato-a -L-idopyranosyluronate)-(1 4)-(2-amino-2-deoxy-3-
O-methyl-6-O-sodium sulphonato-a -D-glucopyranosyl)-(1 4)-(sodium 3-Q-methyl-2-0-
sodium sulphonato-a -L-idopyranosyluronate)-(1^4)-(2-amino-2-deoxy-3-0-methyl-6-0-
sodium sulphonato-g -D-glucopyranosvD-d 4)-(sodium 3-Q-methyl-2-0-sodium
sulphonato-a -L-idopyranosyluronate)-(1^4)-2-amino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-B -D-glucopyranoside (53)
Ammonium formate (27 mg, 0.426 mmol) and 10% palladium-on-carbon (54.5 mg)
are added, under an inert atmosphere, to a solution of compound 52 (21 .8 mg,
0.0085 mmol) in a 1/1 v/v fe/t-butanol/water mixture ( 1 .7 ml). After stirring at ambient
temperature for 3 h 30 min, the reaction medium is filtered (Millipore® LSWP 5 m h filter)
and concentrated to dryness. The residue is loaded onto a column of Sephadex ® G25-fine
gel (95 c 2 cm) eluted with a 0.2 M aqueous solution of NaCI. The fractions containing the
expected compound are combined, and loaded onto a column of Sephadex ® G25-fine gel
(95 x 2 cm) eluted with water. The resulting crude product 53 (17.3 mg) is used as it is in
the next step.
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc': 4.75, IdoUA": 5.24,
Glc'": 5.43, ldoUA lv : 5.26, Glcva : 5.43 ldoUAv l: 5.26, Glcv " : 5.43 and ldoUAv l" : 5.18.
SCHEME 8: Preparation of the disaccharide 55
5-Phenylpentyl (methyl 2-0-acetyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-
(1^4)-6-0-acetyl-2-[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-B-D-glucopyranoside
54)
Compound 42 (519.8 mg, 0.606 mmol) is processed according to the same
procedure as that described for the preparation of 43, to give compound 54 (483.1 mg).
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc': 4.61 ppm and IdollA":
5.03 ppm.
5-Phenylpentyl (methyl 2-0-acetyl-3-0-methyl-a-L-idopyranosyluronate)-(1 ®4)-6-0 -
acetyl-2-[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-B-D-glucopyranoside (55)
Compound 54 (396.4 mg, 0.461 mmol) is processed according to the same
procedure as that described for the preparation of 44, to give compound 55 (383.4 mg).
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc': 4.69 ppm and IdollA":
5.0 ppm.
SCHEME 9: Preparation of the octasaccharide 60
5-Phenylpentyl (methyl 2-0-acetyl-3-0-methyl-4-0-levulinoyl-a -L-idopyranosyluronate)-
( 1®-4)-(6-0-acetyl-2-azido-2-deoxy-3-0-methyl-a -D-glucopyranosyl)-(1 4)-(methyl 2-0-
acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ®-4)-6-0-acetyl-2-
[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-B -D-glucopyranoside (56)
Compound 29 ( 1.17 g, 1.22 mmol) and compound 55 ( 1.50 g, 1.59 mmol) are
processed according to the same procedure as that described for the preparation of 30, to
give compound 56 ( 1 .79 g).
H NMR [500 MHz] (CDCI3) d of the anomeric protons
QI ' b : 4.5, IdollA": 5.00.
5-Phenylpentyl (methyl 2-0-acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ^4)-(6-Qacetyl-
2-azido-2-deoxy-3-0-methyl-a -D-glucopyranosyl)-(1^4)-(methyl 2-Q-acetyl-3-0-
methyl-a -L-idopyranosyluronate)-(1^4)-6-0-acetyl-2-[(benzyloxy)carbonyllamino-2-
deoxy-3-0-methyl-B -D-glucopyranoside (57)
Compound 56 (495 mg, 0.367 mmol) is processed according to the same
procedure as that described for the preparation of 3 1, to give compound 57 (442 mg).
H NMR [500 MHz] (CDCI3) d of the anomeric protons
Glc' b : 4.65 and IdollA": 5.07 and Glc'" a : 4.99 and ldoUA lv : 5.01 .
5-Phenylpentyl (methyl 2-0-acetyl-4-0-levulinoyl-3-0-methyl-a -L-idopyranosyluronate)-
( 1®-4)-(6-0-acetyl-2-azido-2-deoxy-3-0-methyl-a -D-qlucopyranosyl)-(1 4)-(methyl 2-0-
acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ®-4)-(6-0-acetyl-2-azido-2-deoxy-3-0-
methyl-a -D-qlucopyranosyl)-(1 4)-(methyl 2-0-acetyl-3-0-methyl-a -Lidopyranosyluronate)-(
1 ®-4)-6-0-acetyl-2 - (benzyloxy)carbonyllamino-2-deoxy-3-0-
meth l- -D-qlucopyranoside (58)
Compound 57 (433.7 mg, 0.347 mmol) and compound 29 (338 mg, 0.45 mmol)
are processed according to the same procedure as that described for the preparation of
32, to give compound 58 (534 mg).
LC-MS m/z 1860 [(M + Na)+] . TR = 17.02 min
5-Phenylpentyl (methyl 2-0-acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ^4)-(6-0-
acetyl-2-azido-2-deoxy-3-0-methyl-a -D-qlucopyranosyl)-(1^4)-(methyl 2-Q-acetyl-3-0-
methyl-a -L-idopyranosyluronate)-(1^4)-(6-0-acetyl-2-azido-2-deoxy-3-0-methyl-a -Dqlucopyranosyl)-(
1 ®-4)-(methyl 2-0-acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ®-4)-6-
0-acetyl-2-[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-B -D-qlucopyranoside (59)
Compound 58 (447 mg, 0.243 mmol) is processed according to the same
procedure as that described for the preparation of 33, to give compound 59 (386 mg).
LC-MS m/z 1762 [(M + Na)+] TR 1 = 18.32 min
5-Phenylpentyl (methyl 2-0-acetyl-4-0-benzyl-3-0-methyl-a -L-idopyranosyluronate)-
( 1®-4)-(6-0-acetyl-2-azido-2-deoxy-3-0-methyl-a -D-qlucopyranosyl)-(1 4)-(methyl 2-0-
acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ®-4)-(6-0-acetyl-2-azido-2-deoxy-3-0-
methyl-a -D-qlucopyranosyl)-(1^4)-(methyl 2-0-acetyl-3-0-methyl-a -Lidopyranosyluronate)-(
1 ®-4)-(6-0-acetyl-2-azido-2-deoxy-3-0-methyl-a -Dglucopyranosyl)-(
1 4)-(methyl 2-0-acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ®-4)-6-
0-acetyl-2-[(benzyloxy)carbonyllarriino-2-deoxy-3-0-methyl-B -D-glucopyranoside (60)
Compound 59 (95.2 mg, 0.0547 mmol) and compound 28 (52.7 mg, 0.071 mmol)
are processed according to the same procedure as that described for the preparation of
34, to give compound 60 (389 mg).
LC-MS m/z 1180.5 [(M + 2H +CH3CN)2+] TR 1 = 18.27 min
SCHEME 10: Preparation of the octasaccharide 64
5-Phenylpentyl (methyl 3-0-methyl-4-0-benzyl-a -L-idopyranosyluronate)-(1 4)-(2-azido-
2-deoxy-3-0-methyl-a -D-glucopyranosyl)-(1 4)-(methyl 3-O-methyl-a -Lidopyranosyluronate)-(
1 ®-4)-(2-azido-2-deoxy-3-0-methyl-a -D-glucopyranosyl)-(1 ®-4)-
(methyl 3-0-methyl-a -L-idopyranosyluronate)-(1^4)-(2-azido-2-deoxy-3-0-methyl-a -DqlucopyranosvQ-
d 4)-(methyl 3-0-methyl-a -L-idopyranosyluronate)-(1 4)-2-
[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl -B-D-glucopyranoside (61)
A 0.5 M solution of sodium methoxide in methanol (129 m I) is added, at 0°C, under
an argon atmosphere, to a solution of compound 60 (100 mg, 0.0431 mmol) in a 1/1 v/v
dichloromethane/methanol mixture (15.6 ml) containing 3A sieve (54 mg). After magnetic
stirring at 0°C for 4 h 50 min, at ambient temperature for 3 h 50 min and at - 18°C for 15 h,
the reaction medium is neutralized with Dowex® 50WX4 H+ resin. After filtration and
concentration under vacuum, the residue is purified by size exclusion chromatography
(Sephadex ® LH20, 120 c 3 cm, 75/20/5 methanol//V,/V-dimethylformamide/water), to give
88.1 mg of compound 6 1.
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc' b : 4.52, IdoUA": 5.18,
Glc'" a : 5.05 , ldoUA lv : 5.18, Glcv : 5.05, ldoUAv l: 5.18, Glcv " a : 5.05 and
ldoUAv l" : 5.14.
5-Phenylpentyl (methyl 3-0-methyl-4-0-benzyl-2-0-triethylammonium sulphonato -g-Lidopyranosyluronate)-(
1^4)-(2-azido-2-deoxy-3-0-methyl-6-0-triethylammonium
sulphonato -g-D-qlucopyranosyl)-(1 4)-(methyl 3-0-methyl-2-0-triethylammonium
sulphonato-a -L-idopyranosyluronate)-(1 ®-4)-(2-azido-2-deoxy-3-0-methyl-6-0-
triethylammonium sulphonato-a -D-qlucopyranosyl)-(1 4)-(methyl 3-0-methyl-2-0-
triethylammonium sulphonato -g-L-idopyranosyluronate)-(1 4)-(2-azido-2-deoxy-3-0-
methyl-6-O-triethylammonium sulphonato-a -D-qlucopyranosyl)-(1 4)-(methyl 3-O-methyl-
2-O-triethylammonium sulphonato-a -L-idopyranosyluronate)-(1^4)-2-
r(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-6-0-triethylammonium sulphonato- b- -
qlucopyranoside (62)
Compound 6 1 (175 mg, 0.0882 mmol) is processed according to the same
procedure as that described for the preparation of 36, to give compound 62 (229 mg).
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc' b : 4.39, IdoUA": 5.32,
Glc'" a : 5.20, ldoUA lv : 5.32, Glcv : 5.20, ldoUAv l: 5.32, Glcv " a : 5.20 and ldoUAv l" :
5.32.
5-Phenylpentyl (lithium 3-0-methyl-4-0-benzyl-2-0-lithium sulphonato -g-Lidopyranosyluronate)-(
1 4)-(2-azido-2-deoxy-3-0-methyl-6-0-lithium sulphonato -g-Dqlucopyranosyl)-(
1 4)-(lithium 3-Q-methyl-2-0-lithium sulphonato -g-Lidopyranosyluronate)-(
1^4)-(2-azido-2-deoxy-3-0-methyl-6-0-lithium sulphonato -g-Dqlucopyranosyl)-(
1 ^4)-(lithium 3-Q-methyl-2-0-lithium sulphonato -g-Lidopyranosyluronate)-(
1^4)-(2-azido-2-deoxy-3-0-methyl-6-0-lithium sulphonato-g -Dglucopyranosyl)-(
1 4)-(lithium 3-Q-methyl-2-0-lithium sulphonato-g -Lidopyranosyluronate)-(
1^4)-2-[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-6-0-
lithium sulphonato-B -D-qlucopyranoside (63)
Compound 62 (52.1 mg, 0.019 mmol) is processed according to the same
procedure as that described for the preparation of 37, to give compound 63 (44.3 mg).
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc' b : 4.49, IdollA": 5.16,
Glc'" a : 5.30, ldoUA lv : 5.18, Glcv : 5.31 , ldoUAv l: 5.18, Glcv " a : 5.29, and
ldoUAv l" : 5.14.
5-Phenylpentyl (sodium 3-Q-methyl-2-0-sodium sulphonato-a -L-idopyranosyluronate)-
( 1 4)-(2-amino-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a -D-qlucopyranosyl)-(1 4)-
(sodium 3-Q-methyl-2-0-sodium sulphonato-a -L-idopyranosyluronate)-(1 4)-(2-amino-2-
deoxy-3-0-methyl-6-0-sodium sulphonato-a -D-qlucopyranosyl)-(1 4)-(sodium 3-0-
methyl-2-O-sodium sulphonato-a -L-idopyranosyluronate)-(1^4)-(2-amino-2-deoxy-3-0-
methyl-6-O-sodium sulphonato-a -D-qlucopyranosyl)-(1 4)-(sodium 3-Q-methyl-2-0-
sodium sulphonato-a -L-idopyranosyluronate)-(1^4)-2-amino-2-deoxy-3-0-methyl-6-0-
sodium sulphonato-p -D-qlucopyranoside (64)
Compound 63 (43.1 mg, 0.0152 mmol) is processed according to the same
procedure as that described for the preparation of 38, to give compound 64 (31 .6 mg).
ESI-MS m/z 565.07[(M - 4H)4 ] .
SCHEME 11: Preparation of the disaccharide 69
(Methyl 2-0-acetyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1 4)-1 -O-acetyl-
2-azido-2-deoxy-3-0-methyl-a-D-glucopyranose (66)
[ie -Bu2SnCI(OH)] 2 (451 mg, 1.58 mmol), prepared according to A. Orita et al.,
Chem. Eur. J. (2001 ) 7, 3321 , is added, at ambient temperature under an inert
atmosphere, to a solution of compound 65 (7.31 g, 11.28 mmol; WO201 0/0291 85) in 1/1
methanol/tetrahydrofuran (144 ml). After magnetic stirring at 35°C for 5 h, the reaction
medium is concentrated and then the residue is purified by chromatography on a silica gel
column (cyclohexane/acetone), to give 4.33 g of compound 66.
H NMR [500 MHz] (CDCI3) d of the anomeric protons IdoUA": 5.18
Glc' a:6.18.
(Methyl 2-0-acetyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1 4)-1 -O-acetyl-
2-azido-2-deoxy-3-0-methyl-6-0-te/f-butyldiphenylsilyl-g-D-qlucopyranose (67)
Imidazole (109 mg, 1.60 mmol) and ie/f-butyldiphenylsilyl chloride (22 m I,
0.08 mmol) are added, at ambient temperature, to a solution of compound 66 (47.1 mg,
0.077 mmol) in L/,/V-dimethylformamide ( 1 ml). After magnetic stirring at 35°C for 5 h and
at ambient temperature for 17 h, the progression of the reaction is stopped by adding
methanol, and the reaction medium is diluted with dichloromethane and then successively
washed with a 2% potassium hydrogen sulphate solution and a saturated solution of
sodium chloride. The organic phase is dried over sodium sulphate, filtered and then
concentrated under vacuum. The resulting residue is purified by flash chromatography on
a silica gel column (toluene/acetone), to give 62.5 mg of compound 67.
H NMR [500 MHz] (CDCI3) d of the anomeric protons IdoUA": 5.25,
Glc' : 6.16.
(Methyl 2-0-acetyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1^4)-2-azido-2-
deoxy-3-0-methyl-6-0-te/t-butyldiphenylsilyl-a,B-D-glucopyranose (68)
Benzylamine (29.7 ml, 272 mmol) is added, under an argon atmosphere at
ambient temperature, to a solution of compound 67 (6.03 g, 7.14 mmol) in tetrahydrofuran
(293 ml). After magnetic stirring for 14 h, the progression of the reaction is stopped at 0°C
by adding a 1M aqueous solution of hydrochloric acid. The organic phase is washed with
water, dried over sodium sulphate, filtered and then concentrated under vacuum. The
residue is purified by flash chromatography on a silica gel column (toluene/acetone), to
give 3.94 g of compound 68.
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc' b : 4.40, IdoUA": 5.18,
Glc' a : 5.19 and IdoUA": 5.25.
(Methyl 2-0-acetyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1^4)-2-azido-2-
deoxy-3-0-methyl-6-0-te/t-butyldiphenylsilyl-a,B-D-glucopyranose trichloroacetimidate
69)
Compound 68 (4.34 g, 5.41 mmol) is processed according to the same procedure
as that described for the preparation of 28, to give compound 69 (4.36 g).
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc' b : 5.60, Glc' a : 6.37 and
IdoUA": 5.23.
SCHEME 12: Preparation of the disaccharide 72
72
5-Phenylpentyl (methyl 2-0-acetyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-
( 1®-4)-2-[(benzyloxy)carbonyl1amino-2-deoxy-3-0-methyl-p-D-qlucopyranoside (70)
Compound 54 ( 1 .01 g, 1.18 mmol) is processed according to the same procedure
as that described for the preparation of 66, to give compound 70 (917 mg).
LC-MS m/z 840.2 [(M + Na)+] . TR = 9.478 min.
5-Phenylpentyl (methyl 2-0-acetyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-
(1^4)-2-[(benzyloxy)carbonyl1amino-2-deoxy-3-0-methyl-6-0-te/f-butyldiphenylsilyl-B-Dglucopyranoside
(71)
Compound 70 (941 mg, 1.15 mmol) is processed according to the same procedure
as that described for the preparation of 67, to give compound 7 1 ( 1.45 g).
LC-MS m/z 1078.2 [(M + Na)+] . TR = 11.585 min.
5-Phenylpentyl (methyl 2-0-acetyl-3-0-methyl-a-L-idopyranosyluronate)-( 1®-4)-2-
[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-6-0-te/f-butyldiphenylsilyl-B-Dglucopyranoside
(72)
Compound 7 1 ( 1 .45 g, 1.37 mmol) is processed according to the same procedure
as that described for the preparation of 44, to give compound 72 ( 1 .18 g).
LC-MS m/z 980.2 [(M + Na)+] . TR = 11.571 min.
SCHEME 13: Preparation of the octasaccharide 77
5-Phenylpentyl (methyl 2-0-acetyl-3-0-methyl-4-0-levulinoyl-a -L-idopyranosyluronate)-
(1^4)-(2-azido-2-deoxy-3-0-methyl-6-0-te -butyldiphenylsilyl-a-D-glucopyranosyl)-
( 1 4)-(methyl 2-0-acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1®4 -2-
[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-6-0-te/f-butyldiphenylsilyl-B -Dglucopyranoside
(73)
Compound 72 ( 1 .17 g, 1.22 mmol) and compound 69 ( 1 .50 g, 1.59 mmol) are
processed according to the same procedure as that described for the preparation of 30, to
give compound 73 ( 1 .79 g).
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc' b : 4.54 IdollA": 5.17,
Glc'" a : 4.94 and ldoUA lv : 5.23.
5-Phenylpentyl (methyl 2-0-acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ^4)-(2-azido-
2-deoxy-3-0-methyl-6-0-te/t-butyldiphenylsilyl-a -D-glucopyranosyl)-(1^4)-(methyl 2-0-
acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1^4)-2-[(benzyloxy)carbonyllamino-2-
deoxy-3-0-methyl-6-0-te/t-butyldiphenylsilyl-B -D-glucopyranoside (74)
Compound 73 ( 1 .78 g, 1.02 mmol) is processed according to the same procedure
as that described for the preparation of 3 1, to give compound 74 ( 1 .66 g).
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc' b : 4.54, IdollA": 5.17,
Glc'" a : 4.94 and ldoUA lv : 5.23.
5-Phenylpentyl (methyl 2-0-acetyl-3-0-methyl-4-0-levulinoyl-a -L-idopyranosyluronate)-
( 1®-4)-(2-azido-2-deoxy-3-0-methyl-6-0-acetyl-a -D-qlucopyranosyl)-(1 4)-(methyl 2-0-
acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1^4)-(2-azido-2-deoxy-3-0-methyl-6-0-te/fbutyldiphenylsilyl-
a -D-glucopyranosyl)-(1 4)-(methyl 2-0-acetyl-3-0-methyl-a -Lidopyranosyluronate)-(
1^4)-2-[(benzyloxy)carbonyl1amino-2-deoxy-3-0-methyl-6-0-te/fbutyldiphenylsilyl-
p -D-qlucopyranoside (75)
Compound 74 ( 1 .65 g, 1.02 mmol) and compound 29 (980 mg, 1.31 mmol) are
processed according to the same procedure as that described for the preparation of 32, to
give compound 75 ( 1 .75 g).
H NMR [500 MHz] (CDCI3) d of the anomeric protons
Glc' b : 4.54, IdoUA": 5.17, Glc'" a : 4.93, ldoUAlv : 5.30, Glcv a : 5.02 and ldoUAv l:
5.09.
5-Phenylpentyl (methyl 2-0-acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ^4)-(2-azido-
2-deoxy-3-0-methyl-6-0-acetyl-a -D-glucopyranosyl)-(1 ®-4)-(methyl 2-Q-acetyl-3-0-
methyl-a -L-idopyranosyluronate)-(1^4)-(2-azido-2-deoxy-3-0-methyl-6-0-te/fbutyldiphenylsilyl-
a -D-glucopyranosyl)-(1 4)-(methyl 2-0-acetyl-3-0-methyl-a -Lidopyranosyluronate)-(
1^4)-2-[(benzyloxy)carbonyl1amino-2-deoxy-3-0-methyl-6-0-te/fbutyldiphenylsilyl-
B-D-glucopyranoside (76)
Compound 75 ( 1 .74 g, 0.78 mmol) is processed according to the same procedure
as that described for the preparation of 33, to give compound 76 ( 1 .53 g).
H NMR [500 MHz] (CDCI3) d of the anomeric protons
QI ' b : 4.54, IdollA": 5.17, Glc'" a : 4.92, ldoUA lv : 5.30, Glcv : 5.01 and ldoUAv l:
5.01 .
5-Phenylpentyl (methyl 2-0-acetyl-4-0-benzyl-3-0-methyl-a -L-idopyranosyluronate)-
( 1®-4)-(2-azido-2-deoxy-3-0-methyl-6-0-acetyl-a -D-glucopyranosyl)-(1 4)-(methyl 2-0-
acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1^4)-(2-azido-2-deoxy-3-0-methyl-6-0-
acetyl-a -D-glucopyranosyl)-(1 4)-(methyl 2-0-acetyl-3-0-methyl-a -Lidopyranosyluronate)-(
1^4)-(2-azido-2-deoxy-3-0-methyl-6-0-te/f-butyldiphenylsilyl-a -Dglucopyranosyl)-(
1 4)-(methyl 2-0-acetyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ®-4)-2-
[(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-6-0-te/f-butyldiphenylsilyl-B -Dqlucopyranoside
(77)
Compound 76 (200.2 mg, 0.094 mmol) and compound 28 (90.5 mg, 0.122 mmol)
are processed according to the same procedure as that described for the preparation of
34, to give compound 77 (626.3 mg).
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc' b : 4.54 IdoUA": 5.17,
Glc'" a : 4.93, ldoUA lv : 5.29, Glcv : 5.01 , ldoUAv l: 5.10, Glcv " a : 4.99 and ldoUAv l"
5.14.
SCHEME 14: Preparation of the octasaccharide 82
5-Phenylpentyl (methyl 4-0-benzyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ^4)-(2-azido-
2-deoxy-3-0-methyl-a -D-glucopyranosyl)-(1^4)-(methyl 3-O-methyl-a-Lidopyranosyluronate)-(
1®-4)-(2-azido-2-deoxy-3-0-methyl-a-D-glucopyranosyl)-(1®-4)-
(methyl 3-0-methyl-a -L-idopyranosyluronate)-(1^4)-(2-azido-2-deoxy-3-0-methyl-6-0-
te/t-butyldiphenylsilyl-a-D-glucopyranosyl)-(1^4)-(methyl 3-O-methyl-a-Lidopyranosyluronate)-(
1 4)-2-[(benzyloxy)carbonyl1amino-2-deoxy-3-0-m
butyldiphenylsilyl-B-D-glucopyranoside (78)
Compound 77 ( 130 mg, 0.0479 mmol) is processed according to the same
procedure as that described for the preparation of 35, to give compound 78 ( 106 mg).
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc' b : 4.56, IdollA": 5.24,
Glc'" a : 5.06, ldoUAlv : 5.19, Glcv : 5.07, ldoUAv l: 5.20, Glcv " a : 5.00 and ldollA v "
5.11.
5-Phenylpentyl (methyl 4-0-benzyl-3-0-methyl-2-0-triethylammonium sulphonato-g -Lidopyranosyluronate)-(
1 ^4)-(2-azido-2-deoxy-3-0-methyl-6-0-triethylammonium
sulphonato-g -D-glucopyranosyl)-(1 4)-(methyl 3-0-methyl-2-0-triethylammonium
sulphonato-a -L-idopyranosyluronate)-(1 ®-4)-(2-azido-2-deoxy-3-0-methyl-6-0-
triethylammonium sulphonato-a -D-qlucopyranosyl)-(1 4)-(methyl 3-0-methyl-2-0-
triethylammonium sulphonato-a -L-idopyranosyluronate)-(1 ^4)-(2-azido-2-deoxy-3-0-
methyl-6-0-te/f-butyldiphenylsilyl-a -D-qlucopyranosyl)-(1 ^4)-(methyl 3-0-methyl-2-0-
triethylammonium sulphonato-a -L-idopyranosyluronate)-(1 ®4 )-2-
(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-6-0-te/f-butyldiphenylsilyl-p -Dqlucopyranoside
(79)
Compound 78 (205 mg, 0.0833 mmol) is processed according to the same
procedure as that described for the preparation of 36, to give compound 79 (234. 1 mg).
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc' b : 4.3 1, IdoUA": 5.36,
Glc'" a : 5.26, ldoUAlv : 5.39, Glcv : 5.25, ldoUAv l: 5.34, Glcv " a : 5.18 and ldollA v "
5.34.
5-Phenylpentyl (methyl 4-0-benzyl-3-0-methyl-2-0-ammonium sulphonato-g -Lidopyranosyluronate)-(
1 4)-(2-azido-2-deoxy-3-0-methyl-6-0-ammonium sulphonato-g-
D-glucopyranosvD-d 4)-(methyl 3-Q-methyl-2-0-ammonium sulphonato-g -Lidopyranosyluronate)-(
1 ^4)-(2-azido-2-deoxy-3-0-methyl-6-0-ammonium sulphonato-g-
D-glucopyranosvD-d ^4)-(methyl 3-Q-methyl-2-0-ammonium sulphonato-g -Lidopyranosyluronate)-(
1 ®-4)-(2-azido-2-deoxy-3-0-methyl-g -D-glucopyranosyl)-(1 ®-4)-
(rnethyl 3-Q-methyl-2-0-ammonium sulphonato-g -L-idopyranosyluronate)-(1 ®4 )-2-
[(benzyloxy)carbonyl1amino-2-deoxy-3-0-methyl-B -D-glucopyranoside (80)
Ammonium fluoride (221 mg, 80 molar equivalents) is added to a solution of
compound 79 (230 mg, 0.0748 mmol) previously obtained in methanol (9.7 ml). After
magnetic stirring at 55°C for 20 h, the reaction mixture is purified using a Sephadex ® G25-
fine gel column (800 ml) eluted with a 0.2 M aqueous solution of NaCI. The fractions
containing the expected compound are combined , and loaded onto a Sephadex ® G25-fine
gel column (800 ml) eluted with water. The fractions containing the product are then
concentrated under strong vacuum, to give compound 80 ( 195.7 mg).
H NMR [500 MHz] (CD3OD) d of the anomeric protons Glc' b : 4.51 IdollA": 5.26,
Glc'" a : 5.38, ldoUA lv : 5.27, Glcv : 5.38, ldoUAv l: 5.26, Glcv " a : 5.38 and ldoUAv l" :
5.23.
5-Phenylpentyl (lithium 4-0-benzyl-3-0-methyl-2-0-lithium sulphonato-g -Lidopyranosyluronate)-(
1 ^4)-(2-azido-2-deoxy-3-0-methyl-6-0-lithium sulphonato-g -Dqlucopyranosyl)-(
1 4)-(lithium 3-0-methyl-2-0-lithium sulphonato-g -Lidopyranosyluronate)-(
1 ^4)-(2-azido-2-deoxy-3-0-methyl-6-0-lithium sulphonato-g -Dqlucopyranosyl)-(
1 ®-4V(lithium 3-Q-methyl-2-0-lithium sulphonato-g -Lidopyranosyluronate)-(
1 ®-4)-(2-azido-2-deoxy-3-0-methyl-g -D-qlucopyranosyl)-(1 ®-4)-
(lithium 3-Q-methyl-2-0-lithium sulphonato-g -L-idopyranosyluronate)-(1 ®4 )-2-
(benzyloxy)carbonyllamino-2-deoxy-3-0-methyl-p -D-qlucopyranoside (81)
Compound 80 ( 193 mg, 0.0743 mmol) is processed according to the same
procedure as that described for the preparation of 37, to give compound 8 1 ( 178.5 mg).
H NMR [600 MHz] (CD3OD) d of the anomeric protons Glc' b : 4.54, IdoUA": 5.20,
Glc'" a : 5.32, ldoUA lv : 5.24, Glcv : 5.33, ldoUAv l: 5.23, Glcv " a : 5.34 p and
ldoUAv l" : 5.18.
5-Phenylpentyl (sodium 3-Q-methyl-2-0-sodium sulphonato-g -L-idopyranosyluronate)-
( 1 4)-(2-amino-2-deoxy-3-0-methyl-6-0-sodium sulphonato-g -D-qlucopyranosyl)-(1 4 )-
(sodium 3-Q-methyl-2-0-sodium sulphonato-g -L-idopyranosyluronate)-(1 4)-(2-amino-2-
deoxy-3-0-methyl-6-0-sodium sulphonato-g -D-qlucopyranosyl)-(1 4)-(sodium 3-0-
methyl-2-O-sodium sulphonato-g -L-idopyranosyluronate)-(1 4)-(2-amino-2-deoxy-3-0-
methyl-g -D-qlucopyranosvD-d 4)-(sodium 3-Q-methyl-2-0-sodium sulphonato-g -Lidopyranosyluronate)-(
1 ®-4)-2-amino-2-deoxy-3-0-methyl-B -D-qlucopyranoside (82)
Compound 8 1 (23 mg, 0.00875 mmol) is processed according to the same
procedure as that described for the preparation of 38, to give compound 82 ( 16.2 mg).
H NMR [600 MHz] (CD3OD) d of the anomeric protons Glc' b : 4.48, IdoUA": 5.34,
Glc'" a : 5.46, ldoUA lv : 5.31 , Glcv : 5.46, ldoUAv l: 5.26, Glcv " a : 5.46 p and
ldoUAv l" : 5.23.
SCHEME 15: Preparation of the disaccharide 92
9 1
1,6-Anhvdro-2-0-butyl-4-0-tetrahvdropyranyl-p-D-qlucopyranose (84)
Butan-1-ol (16.1 ml, 176 mmol), dropwise, and then 55% sodium hydride (3.5 g,
88 mmol), in several fractions, are successively added at 0°C to a solution of compound
83 (2 g, 8.8 mmol, described in Carbohydrate Research, 64 (1978) 339-364) in ethylene
glycol dimethyl ether (88 ml). At the end of the addition, the temperature is gradually
increased to 85°C and the reaction mixture is stirred magnetically for 5 h 15 min. The
mixture is then diluted at 0°C with ethyl acetate. The organic phase is washed with water,
dried over sodium sulphate, filtered and then evaporated under vacuum. The residue is
purified by chromatography on a silica gel column (toluene/acetone), to give 1.81 g of
compound 84.
H NMR [500 MHz] (CDCI3) d of the anomeric proton: 5.5 Glc'.
1,6-Anhvdro-2-0-butyl-3-0-methyl-4-0-tetrahvdropyranyl-B-D-glucopyranose (85)
55% sodium hydride (400 mg, 10 mmol) is added at 0°C to a solution of compound
84 (2.02 g, 6.7 mmol) in L/,/V-dimethylformamide (67 ml). After stirring at ambient
temperature for 20 min, iodomethane (830 m I, 13.4 mmol) is added dropwise at 0°C. After
stirring at ambient temperature for 1 h, methanol ( 1 .7 ml) is added at 0°C and, after
stirring at ambient temperature for 1 h, the mixture is concentrated under vacuum. The
compound obtained is used in the next step without purification or characterization.
1,6-Anhvdro-2-0-butyl-3-0-methyl-B-D-qlucopyranose (86)
The residue previously obtained is dissolved in methanol (37 ml) and then a 1M
aqueous solution of hydrochloric acid (7.4 ml) is added dropwise at 0°C. After stirring at
ambient temperature for 1 h 30 min, a 1M aqueous solution of sodium hydroxide (7 ml) is
added at 0°C, and then the mixture is concentrated under vacuum. The residue obtained
is purified by chromatography on a silica gel column (toluene/acetone), to give 1.37 g of
compound 86.
SFC-MS m/z 255 [(M + Na)+] . TR = 8.21 min
(2-0-Benzoyl-4,6-0-isopropylidene-3-0-methyl-a-L-idopyranosyl)-(1 ®-4)-1 ,6-anhvdro-2-
0-butyl-3-0-methyl-p-D-qlucopyranose (87)
A mixture of thioglycoside 12 (2.9 g, 7.7 mmol), of the glycosyl acceptor 86 ( 1 .37 g,
5.9 mmol) and of 4 A molecular sieve powder (3.9 g) in dichloromethane (88 ml) is stirred
under an argon atmosphere for 1 h 30 min at ambient temperature. The reaction mixture
is cooled to -20°C and /V-iodosuccinimide ( 1 .85 g, 8.26 mmol), in solution in a 1/1
dioxane/dichloromethane mixture (30 ml), and a 1M solution of triflic acid in a 1/1
dioxane/dichloromethane mixture ( 1.16 ml) are successively added. After stirring for
15 min, the reaction medium is neutralized by adding solid sodium hydrogen carbonate
and then filtered through Celite®. The filtrate is then washed with a saturated solution of
sodium thiosulphate. The organic phase is dried over sodium sulphate, filtered and then
evaporated under vacuum. The residue is purified by chromatography on a silica gel
column (heptane/ethyl acetate), to give 2.36 g of compound 87.
H NMR [500 MHz] (CDCI3) d of the anomeric protons: IdoUA" 5.22 and Glc': 5.38.
(2-0-Benzoyl-3-0-methyl-a-L-idopyranosyl)-( 1®4)-1 ,6-anhydro-2-0-butyl-3-0-methyl-BD-
glucopyranose (88)
Aqueous acetic acid (70%) (8.6 ml) is added at ambient temperature to a solution
of compound 87 (2.36 g, 4.3 mmol) in 1,2-dichloroethane ( 1 .7 ml). After stirring at 60°C for
2 h, the reaction medium is concentrated under vacuum. The residue is coevaporated with
toluene and then purified by chromatography on a silica gel column (toluene/acetone), to
give 2.06 g of compound 88.
H NMR [500 MHz] (CDCI3) d of the anomeric protons: IdollA" 5.21 and Glc': 5.43.
(Methyl 2-0-benzoyl-3-0-methyl-a-L-idopyranosyluronate)-(1 4)-1 ,6-anhydro-2-0-butyl-
3-Q-methyl-B-D-glucopyranose (89)
A solution of 2,2,6, 6-tetramethylpiperidin-1-oxy (13 mg, 0.0804 mmol) in
tetrahydrofuran (270 m I) and a solution of 1,3-dibromo-5,5-dimethylhydantoin (2.3 g,
8.04 mmol) in tetrahydrofuran (6.9 ml) are added successively, at 0°C, to a solution of
compound 88 (2.06 g, 4.02 mmol) in tetrahydrofuran (14.1 ml) and of saturated sodium
hydrogen carbonate (16.1 ml). After stirring at ambient temperature for 3 h 15 min, the
reaction medium is concentrated. The residue is coevaporated with L/,/V-dimethylformamide
and the compound obtained is used in the next step without purification. The
residue obtained is dissolved in L/,/V-dimethylformamide (28 ml) and then solid potassium
hydrogen carbonate (2.0 g) and iodomethane (2.5 ml) are added at 0°C. After magnetic
stirring at ambient temperature for 16 h, the reaction mixture is concentrated. The residue
obtained is dissolved in dichloromethane and is then washed with a saturated aqueous
solution of sodium thiosulphate, dried over sodium sulphate, filtered and then evaporated
under vacuum. A brief purification was carried out (toluene/acetone). Compound 89 was
obtained with sufficient purity to be used in the next step.
H NMR [500 MHz] (CDCI3) d of the anomeric protons Ido: IdoUA" 5.18 and
Glc': 5.32.
(Methyl 2-0-benzoyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1 4)-1 ,6-
anhvdro-2-0-butyl-3-0-methyl-B-D-glucopyranose (90)
4-dimethylaminopyridine (98 mg, 0.804 mmol), 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide hydrochloride ( 1 .5 g, 8.04 mmol) and levulinic acid (827 m I, 8.04 mmol)
are added successively to a solution of compound 89 in dioxane (48.2 ml). After stirring at
ambient temperature for 16 h, the reaction mixture is diluted with dichloromethane. The
organic phase is washed successively with a 10% aqueous solution of potassium
hydrogen sulphate, a saturated aqueous solution of sodium hydrogen carbonate and a
saturated aqueous solution of sodium chloride, and then dried over sodium sulphate,
filtered and evaporated to dryness. A brief purification was carried out (toluene/acetone).
Compound 90 was obtained with sufficient purity to be used in the next step.
H NMR [500 MHz] (CDCI3) d of the anomeric protons Ido: IdollA" 5.31 and
Glc': 5.4.
(Methyl 2-0-benzoyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1 4)-1 ,6-di-Oacetyl-
2-0-butyl-3-0-methyl-g, b-D-glucopyranose (91)
Trifluoroacetic acid (3.5 ml, 44.2 mmol) is added, at 0°C, to a solution of
compound 90 in acetic anhydride (38 ml). The reaction medium is stirred for 16 h at
ambient temperature. After concentration, the mixture is coevaporated with toluene.
Purification of the residue by chromatography on a silica gel column (toluene/acetone)
gives 2.4 g of compound 9 1.
Rf = 0.48, silica gel, 4/1 v/v toluene/acetone
SCHEME 16: Preparation of the disaccharide 97
(Methyl 2-0-benzoyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1^4)-6-0-
acetyl-2-0-butyl-3-0-methyl-a,B-D-glucopyranose (92)
Acetic acid (8.7 m I, 0.15 mmol) and then morpholine (2.7 ml, 30.5 mmol) are added,
at 0°C, to a solution of compound 9 1 (2.26 g, 3.05 mmol) in toluene (6.1 ml). After stirring
at ambient temperature for 6 h 15 min, the progression of the reaction is stopped by
adding, at 0°C, a 1M aqueous solution of hydrochloric acid (31 .5 ml). The aqueous phase
is extracted with ethyl acetate. The combined organic phases are dried over sodium
sulphate, filtered and then concentrated to dryness. The residue is chromatographed on a
silica gel column (toluene/acetone), to give 2.0 g of compound 92.
Rf = 0.26, silica gel, 4/1 v/v toluene/acetone.
(Methyl 2-0-benzoyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1^4)-6-0-
acetyl-2-0-butyl-3-0-methyl-a,B-D-glucopyranose trichloroacetimidate (93)
Trichloroacetonitrile ( 1 .4 ml, 14.3 mmol) and caesium carbonate ( 1.49 g,
4.6 mmol) are added, at 0°C, to a solution of compound 92 (2.0 g, 2.86 mmol) in
dichloromethane (54 ml). After stirring at ambient temperature for 1 h 30 min, the reaction
medium is filtered through Celite® and then concentrated. The residue is purified by
chromatography on a silica gel column (toluene/acetone + 0.1% triethylamine), to give
2.17 g of compound 93.
Rf = 0.46, silica gel, 4/1 v/v toluene/acetone
5-Phenylpentyl (methyl 2-0-benzoyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-
(1®4V6-0-acetyl-2-0-butyl-3-0-methyl-g.B-D-qlucopyranoside (94)
A mixture of trichloroacetimidate 93 (4.73 g, 5.6 mmol), of 5-phenylpentan-1-ol
(4.7 ml, 28 mmol) and of 4 A molecular sieve powder (7.3 g) in dichloromethane (252 ml)
is stirred under an argon atmosphere for 1 h 30 min at ambient temperature. The reaction
mixture is cooled to -20°C and fe/t-butyldimethylsilyl triflate (296 m I, 1.12 mmol) is added
dropwise. After stirring at -20°C for 45 min, the progression of the reaction is stopped by
adding solid sodium hydrogen carbonate. The reaction medium is filtered through Celite®
and then the filtrate is washed with a 2% aqueous solution of sodium hydrogen carbonate.
The organic phase is dried over sodium sulphate, filtered and then evaporated under
vacuum. The residue is purified by chromatography on a silica gel column
(cyclohexane/acetone), to give 4.55 g of compound 94.
H NMR [500 MHz] (CDCI3) d of the anomeric protons IdoUA" 5.19 ppm,
Glc'a: 4.86 ppm and QI 'b: 4.23 ppm.
5-Phenylpentyl (methyl 2-0-benzoyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-
( 1®4)-2-0-butyl-3-0-methyl-B-D-glucopyranoside (95b) and 5-phenylpentyl (methyl 2-0-
benzoyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1^4)-2-0-butyl-3-0-methylg-
D-glucopyranoside (95a)
[tBu2SnCI(0H)] 2 (220 mg, 0.773 mmol), prepared according to A. Orita et al.,
Chem. Eur. J. (2001 ) 7, 3321 , is added, at ambient temperature under an argon
atmosphere, to a solution of compound 94 (4.35 g, 5.15 mmol) in a 1/1
methanol/tetrahydrofuran mixture (62 ml). After magnetic stirring at ambient temperature
for 40 h, the reaction medium is concentrated under vacuum and then the residue is
purified by chromatography on a silica gel column (cyclohexane/acetone), to give 1.44 g
of compound 95b , 1.02 g of compound 95a and 580 mg of 95a b mixture.
Rf (95b) = 0.25 and (95a) 0.13, silica gel, 4/1 v/v diisopropyl ether/ethyl acetate
5-Phenylpentyl (methyl 2-0-benzoyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-
( 1®-4)-2-0-butyl-3-0-methyl-6-0-te/f-butyldiphenylsilyl-B-D-qlucopyranoside (96)
Triethylamine (345 m I, 2.5 mmol), 4-dimethylaminopyridine (61 mg, 0.5 mmol) and
fe/t-butyldiphenylsilyl chloride (520 m I, 2.0 mmol) are added, under an argon atmosphere
at 0°C, to compound 95b (800 mg, 1.0 mmol) dissolved in dichloromethane. The reaction
medium is stirred at ambient temperature for 22 h and then ie/f-butyldiphenylsilyl chloride
(130 m I, 0.5 mmol) is again added. After stirring at ambient temperature for 3 days, the
progression of the reaction is stopped by adding methanol (122 m I, 2.75 mmol). After
magnetic stirring for 30 min, the organic phase is washed with a 10% solution of
potassium hydrogen sulphate, dried over sodium sulphate, filtered and then evaporated
under vacuum. The residue is briefly purified by flash chromatography on a silica gel
column (cyclohexane/acetone + 0.1% triethylamine). Compound 96 is obtained with
sufficient purity to be used in the next step.
Rf= 0.29, silica gel, cyclohexane/acetone 3/1 v/v + 0.1% triethylamine.
5-Phenylpentyl (methyl 2-0-benzoyl-3-0-methyl-a-L-idopyranosyluronate)-(1 ^4)-2-Qbutyl-
3-0-methyl-6-0-te/t-butyldiphenylsilyl-p-D-qlucopyranoside (97)
Hydrazine acetate (460 mg, 5.0 mmol) is added to a solution of compound 96 in a
1/2 toluene/ethanol mixture (200 ml). The reaction medium is stirred for 2 h at ambient
temperature. After concentration under vacuum, the residue is taken up in
dichloromethane and then washed with water. After drying over sodium sulphate, filtration
and then concentration, the residue is chromatographed on a silica gel column
(cyclohexane/acetone + 0.1% triethylamine), to give 850 mg of compound 97.
Rf = 0.28, silica gel, cyclohexane/acetone 3/1 v/v + 0.1% triethylamine.
SCHEME 17: Preparation of the disaccharide 101
10 1
(Methyl 2-0-benzoyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1 4)-1 -Oacetyl-
2-0-butyl-3-0-methyl-g-D-qlucopyranose (98)
[tBu2SnCI(OH)] 2 (610 mg, 2.39 mmol), prepared according to A. Orita et al., Chem.
Eur. J. (2001 ) 7, 3321 , is added to a solution of compound 9 1 ( 1 1.8 g, 15.94 mmol) in a
1/1 methanol/tetrahydrofuran mixture (191 ml). After magnetic stirring at ambient
temperature for 8 h 30 min, the reaction mixture is concentrated under vacuum and then
purified by chromatography on a silica gel column (toluene/acetone), to give compound 98
(8.26 g).
Rf=0.27, silica gel, 4/1 v/v toluene/acetone
(Methyl 2-0-benzoyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1 4)-1 -Oacetyl-
2-0-butyl-3-0-methyl-6-0-te/f-butyldiphenylsilyl-a-D-glucopyranose (99)
Compound 98 (8.26 g, 11.82 mmol) is placed in solution in dichloromethane
(95 ml). 4-dimethylaminopyridine (722 mg, 5.91 mmol), triethylamine (4.1 ml, 29.55 mmol),
and ie/f-butyldiphenylsilyl chloride (6.1 ml, 23.6 mmol) are successively added at 0°C and
under argon. After stirring at ambient temperature for 2 1 h, the progression of the reaction
is stopped by adding methanol ( 1 .2 ml, 26 mmol). After magnetic stirring for 1 h, the
organic phase is washed with a 10% solution of potassium hydrogen sulphate, dried over
sodium sulphate, filtered and then evaporated. The residue is purified by flash
chromatography on a silica gel column (toluene/acetone + 0.1% triethylamine), to give
10.13 g of compound 99.
H NMR [500 MHz] (CD3OD) d of the anomeric protons:
IdoUA" 5.36 and Glc'a 6.28 ppm.
(Methyl 2-0-benzoyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1^4)-2-0-
butyl-3-0-methyl-6-0-te/t-butyldiphenylsilyl-a,B-D-glucopyranose (100)
Acetic acid (9.4 m I, 0.165 mmol) and then morpholine (2.9 ml, 33 mmol) are added,
at 0°C, to a solution of compound 99 (3.1 g, 3.3 mmol) in toluene (6.6 ml). After stirring at
ambient temperature for 24 h, the reaction is stopped by adding, at 0°C, a 1M aqueous
solution of hydrochloric acid (33.6 ml). The aqueous phase is extracted with ethyl acetate.
The combined organic phases are dried over sodium sulphate, filtered and then
concentrated to dryness. The residue is chromatographed on a silica gel column
(toluene/acetone + 0.1% triethylamine), to give 2.7 g of compound 100.
Rf = 0.53 and 0.46, silica gel, toluene/acetone 4/1 v/v + 0.1% triethylamine.
(Methyl 2-0-benzoyl-4-0-levulinoyl-3-0-methyl-a-L-idopyranosyluronate)-(1^4)-2-0-
butyl-3-0-methyl-6-0-te/t-butyldiphenylsilyl-g.p-D-qlucopyranose trichloroacetimidate
(101)
Trichloroacetonitrile ( 1 .5 ml, 15 mmol) and caesium carbonate ( 1 .6 g, 4.8 mmol)
are added, at 0°C, to a solution of compound 100 (2.7 g, 3 mmol) in dichloromethane
(57 ml). After stirring at ambient temperature for 3 h, the reaction medium is filtered
through Celite® and then concentrated under vacuum. The residue is purified by
chromatography on a silica gel column (toluene/acetone + 0.1% triethylamine), to give
3.18 g of compound 101 .
H NMR [500 MHz] (CDCI3) d of the anomeric protons IdoUA": 5.34 ppm,
Glc'a: 6.48 ppm and Glc' b: 5.66 ppm.
SCHEME 18: Preparation of the octasaccharide 106
5-Phenylpentyl (methyl 2-0-benzoyl-4-0-levulinoyl-3-0-methyl-a -L-idopyranosyluronate)-
( 1®-4)-(2-0-butyl-3-0-methyl-6-0-te/f-butyldiphenylsilyl-a-D-glucopyranosyl)-(1 ®-4)-
(methyl 2-0-benzoyl-3-0-methyl-a -L-idopyranosyluronate)-(1^4)-2-0-butyl-3-0-methyl-6-
O-te/f-butyldiphenylsilyl-B -D-glucopyranoside (102)
A mixture of the glycosyl acceptor 97 (850 mg, 0.9 mmol), of compound 101
( 1 .15 g, 1. 1 mmol) and of 4 A molecular sieve powder (825 mg) in dichloromethane
(39 ml) is stirred under an argon atmosphere for 1 h at ambient temperature. The reaction
mixture is cooled to -20°C and fe/t-butyldimethylsilyl triflate (38 m I, 0.165 mmol) is added.
After stirring at -20°C for 1 h, the reaction medium is neutralized by adding solid sodium
hydrogen carbonate, and filtered through Celite®. The filtrate is washed with a 2%
aqueous solution of sodium hydrogen carbonate. The organic phase is dried over sodium
sulphate, filtered and then concentrated under vacuum. The residue obtained is
chromatographed on a silica gel column (cyclohexane/acetone), to give 1.09 g of
compound 102.
Rf = 0.33, silica gel, 3/1 v/v cyclohexane/acetone
5-Phenylpentyl (methyl 2-0-benzoyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ^4)-(2-0-
butyl-3-0-methyl-6-0-te/t-butyldiphenylsilyl-a -D-qlucopyranosyl)-(1^4)-(methyl 2-0-
benzoyl-3-0-methyl-a -L-idopyranosyluronate)-(1^4)-2-0-butyl-3-0-methyl-6-0-te/fbutyldiphenylsilyl-
p -D-qlucopyranoside (103)
Hyrazine acetate (276 mg, 3.0 mmol) is added to a solution of compound 102
( 1 .09 g, 0.6 mmol) in a 1/2 toluene/ethanol mixture (120 ml). The reaction medium is
stirred at ambient temperature for 2 h. After concentration under vacuum, the residue is
placed in solution in dichloromethane and then washed with water. After drying over
sodium sulphate, filtration and then concentration, the residue is purified on a silica gel
column (cyclohexane/acetone), to give 1.02 g of compound 103.
H NMR [600 MHz] (CD3OD) d of the anomeric protons
5.37 ldoUA lv , 4.83 Glc'", 5.35 IdoUA", 4.18 Glc'.
5-Phenylpentyl (methyl 2-0-benzoyl-4-0-levulinoyl-3-0-methyl-a -L-idopyranosyluronate)-
( 1®-4)-(6-0-acetyl-2-0-butyl-3-0-methyl-a -D-glucopyranosyl)-(1 ®-4)-(methyl 2-O-benzoyl-
3-0-methyl-a -L-idopyranosyluronate)-(1 ®-4)-(2-0-butyl-3-0-methyl-6-0-te/fbutyldiphenylsilyl-
a -D-glucopyranosyl)-(1 4)-(methyl 2-0-benzoyl-3-0-methyl-a -Lidopyranosyluronate)-(
1^4)-2-0-butyl-3-0-methyl-6-0-te/f-butyldiphenylsilyl-B -Dglucopyranoside
(104)
A mixture of the glycosyl acceptor 103 ( 1 .02 g, 0.592 mmol), of compound 93
(600 mg, 0.71 mmol) and of 4 A molecular sieve powder (444 mg) in dichloromethane
(20.7 ml) is stirred under an argon atmosphere for 1 h at ambient temperature. The
reaction mixture is then cooled to -20°C and ie/f-butyldimethylsilyl triflate (20.4 m I,
0.089 mmol) is added. After stirring at -20°C for 1 h 15 min , the reaction medium is
neutralized by adding solid sodium hydrogen carbonate, and is then filtered through
Celite®. The filtrate is washed with a 2% aqueous solution of sodium hydrogen carbonate.
The organic phase is dried over sodium sulphate, filtered and then concentrated under
vacuum. The residue obtained is chromatographed on a silica gel column
(cyclohexane/acetone), to give 1.05 g of compound 104.
Rf = 0.31 , silica gel, cyclohexane/acetone 7/3 v/v + 0.1% triethylamine
5-Phenylpentyl (methyl 2-0-benzoyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ®4)-(G-0-
acetyl-2-0-butyl-3-0-methyl-a -D-qlucopyranosyl)-(1 ^4)-(methyl 2-Q-benzoyl-3-0-methyla-
L-idopyranosyluronate)-(1 ^4)-(2-0-butyl-3-0-methyl-6-0-te/f-butyldiphenylsilyl-g -Dqlucopyranosyl)-(
1 4)-(methyl 2-0-benzoyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ®-4 -
2-0-butyl-3-0-methyl-6-0-te/t-butyldiphenylsilyl-p -D-qlucopyranoside (105)
Hydrazine acetate (20 1 mg, 2.18 mmol) is added to a solution of compound 104
( 1 .05 g, 0.44 mmol) in a 1/2 toluene/ethanol mixture (88 ml). After magnetic stirring for
1 h 30 min and concentration, the residue is placed in solution in dichloromethane and
then washed with water. After drying over sodium sulphate, filtration and then
concentration of the organic phase, the residue is purified on a silica gel column
(cyclohexane/acetone), to give 975 mg of compound 105.
H NMR [600 MHz] (CDCI3) d of the anomeric protons: 5.12 ldoUAv l, 4.89 Glcv ,
5.42 ldoUA lv , 4.83 Glc'", 5.34 IdoUA", 4.18 Glc'.
5-Phenylpentyl (methyl 2-0-benzoyl-4-0-levulinoyl-3-0-methyl-a -L-idopyranosyluronate)-
( 1®-4)-(6-0-acetyl-2-0-butyl-3-0-methyl-a -D-qlucopyranosyl)-(1 ®-4)-(methyl 2-O-benzoyl-
3-0-methyl-a -L-idopyranosyluronate)-(1 ^4)-(6-0-acetyl-2-0-butyl-3-0-methyl-a -Dqlucopyranosyl)-(
1 4)-(methyl 2-0-benzoyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ®-4 )-
(2-0-butyl-3-0-methyl-6-0-te/t-butyldiphenylsilyl-a -D-qlucopyranosyl)-(1 ^4)-(methyl 2-0-
benzoyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ^4)-2-0-butyl-3-0-methyl-6-0-te/f-butyldiphenylsilyl-
B -D-glucopyranoside (1 06)
A mixture of the glycosyl acceptor 105 (975 mg, 0.423 mmol), of compound 93
(535 mg, 0.634 mmol) and of 4 A molecular sieve powder (3 17 mg) in toluene ( 15 ml) is
stirred under an argon atmosphere for 1 h at ambient temperature. The reaction mixture is
cooled to -20°C and iert-butyldimethylsilyl triflate ( 15 m I, 0.089 mmol) is added. After
stirring at -20°C for 35 min, the reaction medium is neutralized by adding solid sodium
hydrogen carbonate and filtered through Celite®. The filtrate is washed with a 2% aqueous
solution of sodium hydrogen carbonate. The organic phase is dried over sodium sulphate,
filtered and then concentrated. The residue obtained is chromatographed on a silica gel
column (cyclohexane/acetone), to give 1.25 g of compound 106.
Rf = 0.24, silica gel, 7/3 v/v cyclohexane/acetone
SCHEME 19: Preparation of the octasaccharide 11 1
5-Phenylpentvl (methvl 2-Q-benzoyl-3-0-methyl-a-L-idopyranosyluronateVn ®4Vffi-Qacetyl-
2-0 -butvl-3-0-methvl-a-D-alucopvr anosylV(1®4V(methyl 2-Q-henzoyl-3-0-methyla-
L-idopyranosyluronate)-(1^4)-(6-0-acetyl-2-0-butyl-3-0-methyl-a-D-aluconyrann l)-
( 1 4)-(methyl 2-0-benzoyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ®-4)-(2-0-butyl-3-0-
methyl-6-0-te/f-butyldiphenylsilyl-a -D-glucopyranosyl)-(1^4)-(methyl 2-Q-benzoyl-3-0-
methyl-a -L-idopyranosyluronate)-(1^4)-2-0-butyl-3-0-methyl-6-0-te/f-butyldiphenylsi
b-D-glucopyranoside (107)
Hydrazine acetate (193 mg, 2.09 mmol) is added to a solution of compound 106
( 1 .25 g, 0.419 mmol) in a 1/2 toluene/ethanol mixture (84 ml). The reaction medium is
stirred at ambient temperature for 1 h 45 min. After concentration, the residue is taken up
in dichloromethane and then washed with water. After drying over sodium sulphate,
filtration and then concentration, the residue is purified on a silica gel column
(cyclohexane/acetone), to give 0.99 g of compound 107.
SFC-MS m/z 1463 [(M + 2H + CH3CN)2+]/2. TR 1 = 9.23 min
5-Phenylpentyl (methyl 2-0-benzoyl-3-0-methyl-4-0-te/f-butyldiphenylsilyl-a -Lidopyranosyluronate)-(
1 ®-4)-(6-0-acetyl-2-0-butyl-3-0-methyl-a -D-qlucopyranosyl)-
( 1 4)-(methyl 2-0-benzoyl-3-0-methyl-a -L-idopyranosyluronate)-(1 4)-(6-0-acetyl-2-0-
butyl-3-0-methyl-a -D-qlucopyranosyl)-(1 4)-(methyl 2-0-benzoyl-3-0-methyl-a -Lidopyranosyluronate)-(
1^4)-(2-0-butyl-3-0-methyl-6-0-te/f-butyldiphenylsilyl-a -Dqlucopyranosyl)-(
1 4)-(methyl 2-0-benzoyl-3-0-methyl-a -L-idopyranosyluronate)-(1 ®-4)-
2-0-butyl-3-0-methyl-6-0-te/t-butyldiphenylsilyl-p -D-qlucopyranoside (108)
Imidazole ( 1.3 g, 19.25 mmol) and ie/f-butyldiphenylsilyl chloride (2.5 ml,
9.52 mmol) are successively added, at ambient temperature and under an argon
atmosphere, to a solution of compound 107 ( 1 .09 g, 0.35 mmol) in L/,/V-dimethylformamide
(4.2 ml). After stirring at 60°C for 22 h, the progression of the reaction is
stopped by adding methanol (425 m I, 10.47 mmol). The organic phase is washed with a
10% aqueous solution of potassium hydrogen sulphate, dried over sodium sulphate,
filtered and then evaporated under vacuum. The residue is purified by flash
chromatography on a silica gel column (cyclohexane/acetone), to give 860 mg of
compound 108.
SFC-MS m/z 1582 [(M + 2H + CH3CN)2+]/2. TR 1 = 8.52 min
5-Phenylpentyl (methyl 3-0-methyl-4-0-te/f-butyldiphenylsilyl-a -L-idopyranosyluronate)-
( 1®-4)-(2-0-butyl-3-0-methyl-a -D-qlucopyranosyl)-(1 ®-4)-(methyl 3-O-methyl-a -Lidopyranosyluronate)-(
1 4)-(2-0-butyl-3-0-methyl-a -D-glucopyranosyl)-(1 4)-(methyl 3-
0-methyl-a -L-idopyranosyluronate)-(1^4)-(2-0-butyl-3-0-methyl-6-0-te/fbutyldiphenylsilyl-
a -D-glucopyranosyl)-(1 4)-(methyl 3-O-methyl-q -Lidopyranosyluronate)-(
1^4)-2-0-butyl-3-0-methyl-6-0-te/f-butyldiphenylsilyl-B -Dglucopyranoside
(109)
Potassium fe/t-butoxide (4.1 mg, 0.034 mmol) is added, at 0°C under an argon
atmosphere, to a solution of compound 108 (350 mg, 0.1 12 mmol) in 1/1
methanol/dioxane (728 m I) . After stirring at 0°C for 77 h, the reaction medium is
neutralized by adding Dowex AG 50 WX4 H+ resin, filtered and then concentrated. The
residue is chromatographed on a silica gel column (diisopropyl ether/acetone). The
mixture obtained is again processed under the conditions described above (1/1
methanol/dioxane (460 m I) , potassium ie/f-butoxide (2.4 mg, 0.021 1 mmol), stirring at 0°C
for 48 h, then neutralization by adding Dowex AG 50 WX4 H+ resin, filtration and then
concentration). The residue is chromatographed on a silica gel column (diisopropyl
ether/acetone), to give 147 mg of compound 109.
SFC-MS m/z 1332 [(M + 2H + CH3CN)2+]/2. TR = 10.58 min.
5-Phenylpentyl (methyl 3-0-methyl-4-0-te/f-butyldiphenylsilyl-2-0-triethylammonium
sulphonato-a -L-idopyranosyluronate)-(1^4)-(2-0-butyl-3-0-methyl-6-0-triethylammonium
sulphonato-a -D-qlucopyranosyl)-(1 4)-(methyl 3-0-methyl-2-0-triethylammonium
sulphonato-a -L-idopyranosyluronate)-(1^4)-(2-0-butyl-3-0-methyl-6-0-triethylammonium
sulphonato-a -D-qlucopyranosyl)-(1 4)-(methyl 3-0-methyl-2-0-triethylammonium
sulphonato-a -L-idopyranosyluronate)-(1 ®-4)-(2-0-butyl-3-0-methyl-6-0-te/fbutyldiphenylsilyl-
a -D-qlucopyranosyl)-(1 4)-(methyl 3-0-methyl-2-0-triethylammonium
sulphonato-a -L-idopyranosyluronate)-(1^4)-2-0-butyl-3-0-methyl-6-0-te/fbutyldiphenylsilyl-
p -D-qlucopyranoside (1 10)
Compound 109 (147 mg, 0.056 mmol) is codistilled with anhydrous N,Ndimethylformamide
(3 x 5 ml) and is then placed in solution in anhydrous N,Ndimethylformamide
(5 ml). The sulphur trioxide-triethylamine complex (304 mg,
1.68 mmol) is added to this solution. The mixture is stirred at 55°C for 16 h in the dark and
then the excess reagent is destroyed with methanol (273 m I, 4.98 mmol). The reaction
medium is loaded onto a Sephadex ® LH20 gel column (95 c 2 cm) eluted with a 9/1 v/v
methanol//V,/V-dimethylformamide mixture, to give compound 110 (172 mg).
H NMR [600 MHz] (CD3OD) d of the anomeric protons: 5.38 ldoUA v l" , 5.24 Glcv " ,
5.34 ldoUA v l, 5.29 Glcv , 5.36 ldoUA lv , 5.31 Glc'", 5.38 IdoUA", 4.25 Glc'.
5-Phenylpentyl (methyl 3-Q-methyl-2-0-ammonium sulphonato-a -L-idopyranosyluronate)-
( 1 4)-(2-0-butyl-3-0-methyl-6-0-ammonium sulphonato-a-D-glucopyranosyl)-(1 4)-
(methyl 3-0-methyl-2-0-ammonium sulphonato-g-L-idopyranosyluronate)-(1 4)-(2-0-
butyl-3-0-methyl-6-0-ammonium sulphonato-a-D-glucopyranosyl)-(1 4)-(methyl 3-0-
methyl-2-O-ammonium sulphonato-g-L-idopyranosyluronate)-(1 4)-(2-0-butyl-3-0-
methyl-g-D-glucopyranosvD-d 4)-(methyl 3-Q-methyl-2-0-ammonium sulphonato-g-LidopyranosyluronateVd
4)-2-0-butyl-3-0-methyl-B-D-glucopyranoside (1 11
Ammonium fluoride (77 mg, 2.07 mmol) is added, under argon, to a solution of
compound 110 (44.8 mg, 0.0 172 mmol) in methanol (2.2 ml). After stirring at 55°C for 24 h,
the reaction medium is loaded onto a Sephadex ® LH20 gel column (95 c 2 cm) eluted with
9/1 v/v methanol//V,/V-dimethylformamide, to give compound 111 (38 mg).
H NMR [600 MHz] (CD3OD) d of the anomeric protons: 5.32 ldoUAv l" , 5.29 Glcv " ,
5.37 ldoUAv l, 5.28 Glcv , 5.26 ldoUAlv , 5.33 Glc'", 5.28 IdoUA", 4.28 Glc'.
Examples of compounds according to the invention:
EXAMPLE 1: Methyl (sodium 3-0-methyl-2-0-sodium sulphonato-g-Lidopyranosyluronate)-(
1 ^4)-(2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-g-
D-qlucopyranosyl)-(1 4H(sodium 3-Q-methyl-2-0-sodium sulphonato-g-Lidopyranosyluronate)-(
1 ^4)-(2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-g-
D-qlucopyranosyl)-(1 4) l -(sodium 3-Q-methyl-2-0-sodium sulphonato-g-Lidopyranosyluronate)-(
1 ^4)-2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-g-
D-qlucopyranoside (compound No. 1)
Sodium hydrogen carbonate (84 mg, 1 mmol) and then acetic anhydride (47 m I,
0.5 mmol) are added , at 0°C under an argon atmosphere, to compound 38 ( 15 mg,
0.0063 mmol) dissolved in a saturated aqueous solution of sodium hydrogen carbonate
(8 13 m I) . After stirring at 0°C for 3 h and at ambient temperature for 14 h, the reaction
medium is loaded onto a Sephadex® G25-fine gel column (95 c 2 cm) eluted with a 0.2 M
aqueous solution of NaCI. The fractions containing the expected compound are combined,
and loaded onto a Sephadex® G25-fine gel column (95 c 2 cm) eluted with water.
Compound 1 ( 12 mg) is obtained ( 12 mg) after concentration under vacuum.
H NMR [600 MHz] (D20 ) d of the anomeric protons: 5.15 ldoUA v l" , 5.10 Glcv " ,
5.17 ldoUA v l, 5.1 1 Glcv , 5.16 ldoUA lv , 5.1 1 Glc'", 5.17 IdoUA", 4.72 Glc'.
ESI-MS m/z 574.06 [(M - 4H)4 ]
CE: TR= 4.70 min.
EXAMPLE 2: Pentyl (sodium 3-Q-methyl-2-0-sodium sulphonato-g -Lidopyranosyluronate)-(
1 4)-(2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-g-
D-glucopyranosvD-d 4)-[(sodium 3-Q-methyl-2-0-sodium sulphonato-g -Lidopyranosyluronate)-(
1^4)-(2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-g-
D-glucopyranosvD-d 4)]?-(sodium 3-0-methyl-2-0-sodium sulphonato-g -Lidopyranosyluronate)-(
1^4)-2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato- b-
D-qlucopyranoside (compound No. 2)
Sodium hydrogen carbonate (48 mg, 0.567 mmol) and then acetic anhydride (27 m I,
0.284 mmol) are added, at 0°C under an argon atmosphere, to compound 53 (8.7 mg,
0.00355 mmol) dissolved in a saturated aqueous solution of sodium hydrogen carbonate
(355 m I) . After stirring at ambient temperature for 16 h, the reaction medium is loaded onto
a Sephadex ® G25-fine gel column (95 c 2 cm) eluted with a 0.2 M aqueous solution of
NaCI. The fractions containing the expected compound are combined, and loaded onto a
Sephadex ® G25-fine gel column (95x2 cm) eluted with water. The residue obtained is
reprocessed under the same conditions, to give 9.2 mg of compound 2.
H NMR [600 MHz] (D20 ) d of the anomeric protons: 5.15 ldoUA v l" , 5.09 Glcv " ,
5.15 ldoUA v l, 5.09 Glcv , 5.15 ldoUA lv , 5.09 Glc'", 5.15 IdoUA", 4.54 Glc'.
ESI-MS m/z 588.06 [(M - 4H)4 ]
CE: TR=4.69 min.
EXAMPLE 3: Pentyl (sodium 3-0-methyl-2-0-sodium sulphonato-g -Lidopyranosyluronate)-(
1^4)-(2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-g -D-glucopyranosyl)-(1 4)-[(sodium 3-Q-methyl-2-0-sodium sulphonato-g -Lidopyranosyluronate)-(
1^4)-(2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-g -D-glucopyranosvD-d 4)1?-(sodium 3-0-methyl-2-0-sodium sulphonato-g -Lidopyranosyluronate)-(
1 4)-2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-B -D-glucopyranoside (compound No. 3)
A solution of L/,/V-diisopropylethylamine ( 1 1 m I, 0.063 mmol) in L/,/V-dimethylformamide
(33 m I) and /V-butyroxysuccinimide (9 mg, 0.048 mmol), prepared according to
Naito et al. Journal of Antibiotics, 29; 1976, 1286, dissolved in L/,/V-dimethylformamide
(33 m I) , are added, at 0°C under an argon atmosphere, to compound 53 (8.9 mg,
0.00363 mmol) dissolved in L/,/V-dimethylformamide (471 m I) and water (290 m I) . After
magnetic stirring at ambient temperature for 3.5 h, two further additions of reagents are
carried out (same amounts), 3 h 30 min apart. After stirring at ambient temperature for
16 h, the reaction medium is loaded onto a Sephadex ® G25-fine gel column (95 c 2 cm)
eluted with a 0.2 M aqueous solution of NaCI. The fractions containing the expected
compound are combined, and loaded onto a Sephadex ® G25-fine gel column (95 c 2 cm)
eluted with water. The mixture obtained is reacted under the same conditions, to give
10.3 mg of compound 3.
H NMR [500 MHz] (D20 ) d of the anomeric protons Glc' b : 4.45 ppm, IdoUA": 5.06
ppm, Glc'" a : 4.98 ppm, ldoUA lv : 5.06 ppm, Glcv : 4.98 ppm, ldoUAv l: 5.06 ppm, Glcv " a :
4.98 ppm and ldoUAv l" : 5.05 ppm.
ESI-MS m/z 588.06 [(M - 4H)4 ] .
EXAMPLE 4: 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato-g -Lidopyranosyluronate)-(
1^4)-(2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-g-
D-glucopyranosyl)-(1 4)-[(sodium 3-0-methyl-2-0-sodium sulphonato-g -Lidopyranosyluronate)-(
1^4)-(2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-g-
D-glucopyranosyl-d 4)l?-(sodium 3-0-methyl-2-0-sodium sulphonato-g -Lidopyranosyluronate)-(
1 4)-2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-BD-
glucopyranoside (compound No. 4)
Compound 64 (6.0 mg, 0.0024 mmol) is processed according to the same
procedure as that described for the preparation of Example 1, to give compound 4
(6.3 mg).
H NMR [500 MHz] (D20 ) d of the anomeric protons Glc' b : 4.59 ppm, IdollA": 5.20
ppm, Glc'" a : 5.15 ppm, ldoUA lv : 5.20 ppm, Glcv : 5.15 ppm, ldoUAv l: 5.21 ppm, Glcv "a :
5.15 ppm and ldoUAv l" : 5.19 ppm.
ESI-MS m/z 494.25[(M - 5H)5 ] .
EXAMPLE 5: 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato-g -Lidopyranosyluronate)-(
1^4)-(2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-g -D-qlucopyranosvD-d 4H(sodium 3-Q-methyl-2-0-sodium sulphonato-g -Lidopyranosyluronate)-(
1^4)-(2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-g -D-qlucopyranosyl)-(1 4)1 (sodium 3-Q-methyl-2-0-sodium sulphonato-g -Lidopyranosyluronate)-(
1^4)-2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-p -D-qlucopyranoside (compound No. 5)
Compound 64 (25.8 mg, 0.0102 mmol) is processed according to the same
procedure as that described for the preparation of Example 3, to give compound 5
(23.7 mg).
H NMR [500 MHz] (CDCI3) d of the anomeric protons Glc' b : 4.51 ppm, IdoUA":
5.18 ppm, Glc'" a : 5.07 ppm, ldoUA lv : 5.19 ppm, Glcv : 5.07 ppm, ldoUAv l: 5.20 ppm,
Glcv " a : 5.07 ppm and ldoUAv l" : 5.15 ppm.
ESI-MS m/z 507.88[(M - 5H)5 ] .
EXAMPLE 6: 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato-g-Lidopyranosyluronate)-(
1^4)-(2-butanoylarriino-2-deoxy-3-0-methyl-6-0-sodiurri
sulphonato-g-D-glucopyranosyl)-(1 4)-(sodium 3-Q-methyl-2-0-sodium sulphonato-g-Lidopyranosyluronate)-(
1^4)-(2-butanoylarriino-2-deoxy-3-0-methyl-6-0-sodiurri
sulphonato-g-D-glucopyranosyl)-(1 4)-(sodium 3-0-methyl-2-0-sodium sulphonato-g-Lidopyranosyluronate)-(
1^4)-(2-butanoylarriino-2-deoxy-3-0-methyl-g-D-glucopyranoside)-
( 1 4)-(sodium 3-Q-methyl-2-0-sodium sulphonato-g-L-idopyranosyluronate)-(1 4)-2-
(butanoylamino)-2-deoxy-3-0-methyl-B-D-glucopyranoside (compound No. 6)
Compound 82 (55.5 mg, 0.0239 mmol) is processed according to the same
procedure as that described for the preparation of Example 1, to give compound 6
(66.2 mg).
H NMR [600 MHz] (D20 ) d of the anomeric protons Glc' b : 4.58 IdollA": 5.23, Glc'
a : 5.10, ldoUAlv : 5.24, Glc v a : 5.10, ldoUAv l: 5.23, Glcv " a : 5.13 and ldoUAv l" : 5.20
CE: TR=3.40 min.
EXAMPLE 7: 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato-g-Lidopyranosyluronate)-(
1^4)-(2 - (3-methylbutanoyl)aminol-2-deoxy-3-0-methyl-6-0-
sodium sulphonato-g-D-glucopyranosyl)-(1 4)-(sodium 3-Q-methyl-2-0-sodium
sulphonato-g-L-idopyranosyluronate)-(1 ®-4)-(2-[(3-methylbutanoyl)aminol-2-deoxy-3-0-
methyl-6-O-sodium sulphonato-g-D-glucopyranosyl)-(1 4)-(sodium 3-0-methyl-2-0-
sodium sulphonato-g-L-idopyranosyluronate)-(1^4)-(2-[(3-methylbutanoyl)aminol-2-
deoxy-3-0-methyl-g-D-glucopyranoside)-(1 4)-(sodium 3-Q-methyl-2-0-sodium
sulphonato-g-L-idopyranosyluronate)-(1 ®-4)-2-[(3-methylbutanoyl)aminol-2-deoxy-3-0-
methyl-B-D-glucopyranoside (compound No. 7)
Compound 82 (49.8 mg, 0.0214 mmol) is processed according to the same
procedure as that described for the preparation of Example 1, to give compound 7
(34.0 mg).
H NMR [600 MHz] (D20 ) d of the anomeric protons Glc' b : 4.57, IdollA": 5.23,
Glc'" a : 5.09, ldoUA lv : 5.27, Glcv : 5.09, ldoUAv l: 5.25, Glcv " a : 5.12 and ldoUAv l" : 5.21 .
ESI-MS m/z 642.32 [(M - 4H)4 ] .
EXAMPLE 8: 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato -g-Lidopyranosyluronate)-(
1 4)-(2-0-butyl-3-0-methyl-6-0-sodium sulphonato -g-Dqlucopyranosyl)-(
1 4)-(sodium 3-Q-methyl-2-0-sodium sulphonato -g-Lidopyranosyluronate)-(
1 4)-(2-0-butyl-3-0-methyl-6-0-sodium sulphonato -g-Dqlucopyranosyl)-(
1 4)-(sodium 3-Q-methyl-2-0-sodium sulphonato -g-LidopyranosyluronateVd
4)-(2-0-butyl-3-0-methyl -g-D-qlucopyranosyl)-(1 4)-(sodium 3-
O-methyl-2-O-sodium sulphonato -g-L-idopyranosyluronate)-(1^4)-2-0-butyl-3-0-methyl-
b-D-qlucopyranoside (compound No. 8)
A 4.2 M aqueous solution of lithium hydroxide (290 m I, 1.216 mmol) and a 30%
hydrogen peroxide solution (373 m I, 3.648 mmol) are added, at 0°C under argon, to
compound 1 11 (38 mg, 0.0152 mmol) dissolved in water (555 m I) . After stirring at 0°C for
2 h, at ambient temperature for 17 h and at 45°C for 24 h, the reaction medium is loaded
onto a Sephadex ® G-25 fine column (90 c 3 cm) eluted with a 0.2 M aqueous solution of
sodium chloride. The fractions containing the product are concentrated and desalified
using the same column eluted with water. After concentration to dryness, compound 8
(33 mg) is obtained.
H NMR [600 MHz] (CD 3OD) d of the anomeric protons: 5.17 ldoUA v l" , 5.41 Glc
5.17 ldo UUAAvvil
, 55..4400 GGllccv
, 55..1188 ldoUA'\ 5.40 Glc"', 5.15 IdoUA", 4.50 Glc'
[a] D 14° (c O.25, H20).
EXAMPLE 9 : 5-phenylpentyl (sodium 3-Q-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1 4)-(2-0-butyl-3-0-methyl-6-0-sodium sulphonato -g -Dglucopyranosyl)-(
1 4)-(sodium 3-Q-methyl-2-0-sodium sulphonato -g -Lidopyranosyluronate)-(
1 4)-(2-0-butyl-3-0-methyl-6-0-sodium sulphonato -g -Dglucopyranosyl)-(
1 4)-(sodium 3-Q-methyl-2-0-sodium sulphonato - -Lidopyranosyluronate)-(
1 ®-4)-(2-0-butyl-3-0-methyl -g -D-glucopyranosyl)-(1 ®-4)-(sodium
3-Q-methyl-2-0-sodium sulphonato -g -L-idopyranosyluronate)-(1 4)-2-0-butyl-3-0-
methyl -g -D-qlucopyranoside (compound No. 9)
A n analogous sequence was performed using compound 95a to give compound 9.
H NMR [600 MHz] (CD 3OD) d of the anomeric protons: 5.17 ldoUA v l" , 5.41 Glc v " ,
5.15 ldoUA v l, 5.39 Glc v , 5.17 ldoUA lv , 5.40 Glc'", 5.15 IdoUA", 5.09 Glc'
[g]D 58° (c O.16, H20).
in vitro Anqioqenesis model: specific activity towards FGF2
The in vitro angiogenesis model corresponds to a rearrangement of human vein
endothelial cells o n a biological matrix. The matrix is prepared by distributing, into each
well of a 96-well plate (Becton Dickinson 353872), 6 0 m I of Matrigel ® diluted to 1/3 (Growth
factor reduced Matrigel ® : Becton Dickinson 356230) in collagen (rat Tail collagen, type I:
Becton Dickinson 354249). The biological matrix hardens after 1 hour at 37°C.
Human vein endothelial cells (HUVEC ref: C-12200 - Promocell) are seeded onto
the biological matrix at 7800 cells/well in 120 m I of EBM ® medium (Endothelial Basal
Medium, Lonza C3121) + 2% FCS (foetal calf serum - Lonza) + 10 mg/ml hEGF
(Recombinant Human Epidermal Growth Factor - Lonza). The cells are stimulated with
10 ng/ml FGF2 (R&D Systems/234 - FSE - 0 50) o r with the products of the invention for
18 hours a t 37°C in the presence of 5% C0 2 . After 2 4 hours, the cells are observed under
a microscope (c4 objective) and the length of the pseudo-tubules is analysed using image
software (Biocom Visiolab 2000 software).
In this in vitro angiogenesis test, the compounds of the invention mostly exhibit a
specific activity of between 10 6 M and 10 0 M. For example, compounds No. 4 and 6 are
active at 10 0 M.
Model of cellulose implant in mice
This model is an adaptation of the model described by Andrade et al.
(Microvascular Research, 1997, 54, 253-61) for testing pharmacological products capable
of activating the onset of angiogenesis.
The animals (white inbred BALB/c J mice) are anaesthetized with a xylazine
(Rompun®, 10 mg/kg)/ketamine (Imalgene® 1000, 100 mg/kg) mixture intraperitoneally.
The back of the animal is shaved and disinfected with Hexomedine ®. A pocket of air is
created subcutaneously on the back of the mouse, by injecting 5 ml of sterile air. An
incision of approximately 2 cm, on the top of the back of the animal is made in order to
introduce a sterile cellulose implant (disc 1 cm in diameter, 2 mm thick, Cellspon®
ref. 0501) impregnated with 50 m I of sterile solution containing the test product. The
incision is then sutured and cleaned with Hexomedine®.
On the days following the insertion of the implant, the mice can receive the product
into the implant via an injection through the skin (50 m I/implant/day) under gas
anaesthesia (5% isoflurane (Aerrane®, Baxter)).
Seven days after the insertion of the sponge, the mice are sacrificed by means of
a lethal dose of sodium pentobarbital (CEVA Sante Animale), administered
intraperitoneally. The skin is then excised, approximately 1 cm around the sponge, while
avoiding the scar, so as to release the skin and the sponge. The sponge is then cut into
several pieces and placed in a Ribolyser® tube containing 1 ml of lysis buffer (Cell Death
Detection ELISA, Roche). The tubes are shaken 4 times consecutively, for 20 seconds, at
force 4, using a cell mill (FastPrep® FP 120). The tubes are then centrifuged for
10 minutes at 2000 g at 20°C and the supernatants are frozen at -20°C until the time of
the haemoglobin assay. On the day of the assay, the tubes are again centrifuged after
thawing and the haemoglobin concentration is measured with the Drabkin reagent (Sigma,
volume per volume) by reading on a spectrophotometer at 405 nm against a standard
range of bovine haemoglobin (Sigma).
The haemoglobin concentration in each sample is expressed in mg/ml according
to the polynomial regression produced from the range. The results are expressed as a
mean value (± sem) for each group. The differences between the groups are tested with
an ANOVA followed by a Dunnett test on the square root of the values.
In this in vivo test, the compounds of the invention that were tested demonstrated
a specific activity at 45 ng/site. For example, compounds No. 1 and 3 are active at
45 ng/site.
Thus, the compounds according to the invention increase the formation of new
vessels in vitro and in vivo and post-ischaemic revascularization. The compounds
according to the invention can therefore be used for the preparation of medicaments that
are of use for the treatment of diseases requiring activation of FGF receptors and more
generally in pathological conditions requiring activation of angiogenesis, such as
cicatrisation or post-ischaemic revascularization.
According to another of its aspects, a subject of the invention is therefore
medicaments which comprise a compound of formula (I) according to the invention, or a
pharmaceutically acceptable salt thereof.
These medicaments find their use in therapy, in the treatment of ischaemia
(cardiac ischaemia, lower limb ischaemia), the treatment of diseases associated with
narrowing or obstruction of the arteries or arteritis, the treatment of angina pectoris, the
treatment of thromboangiitis obliterans, the treatment of atherosclerosis, and cicatrisation.
It is also possible to envisage the use of the compounds of the invention for the treatment
of post-angioplasty or post-endarterectomy restenosis; for these pathological conditions,
the use of stents impregnated with the compounds of the invention can be envisaged.
FGFs have been shown to be protective factors in a certain number of pathological
conditions such as: chronic ulcer and refractory ulcer in diabetic or nondiabetic patients,
chronic or nonchronic perforations of the eardrum, periodontitis, muscle regeneration and
myoblast survival, peripheral neuropathy, post-operative nerve damage, nerve
deficiencies such as Parkinson's disease, Alzheimer's disease, prion disease and
neuronal degeneration in alcoholics, dementia, bioartificial pancreas graft survival in
diabetic patients, retinal degeneration, stromal keratitis, pigmentary retinitis, osteoarthritis,
pre-eclampsia, vascular lesions and acute respiratory distress syndrome, post-traumatic
cartilage and bone repair, the repair and protection of hair follicles, and the protection and
regulation of hair growth.
Thus, a subject of the invention is the compounds of formula (I) defined above, for
use thereof in the treatment of the pathological conditions described above.
A subject of the invention is also the use of the compounds of formula (I) defined
above, for the production of a medicament intended for the treatment of the pathological
conditions described above.
According to another of its aspects, the present invention relates to pharmaceutical
compositions comprising, as active ingredient, a compound according to the invention.
These pharmaceutical compositions contain an effective dose of at least one compound
according to the invention, or a pharmaceutically acceptable salt of said compound, and
also at least one pharmaceutically acceptable excipient. Said excipients are chosen
according to the pharmaceutical form and the method of administration desired, from the
usual excipients that are known to those skilled in the art.
In the pharmaceutical compositions of the present invention for oral, sublingual,
subcutaneous, intramuscular, intravenous, topical, local, intratracheal, intranasal,
transdermal or rectal administration, the active ingredient of formula (I) above or salt
thereof can be administered in a unit administration form, as a mixture with conventional
pharmaceutical excipients, to animals and to human beings, for the prevention or
treatment of the above disorders or diseases.
The appropriate unit administration forms include oral forms such as tablets, soft
or hard gel capsules, powders, granules and oral solutions or suspensions, sublingual,
buccal, intratracheal, intraocular or intranasal administration forms, forms for
administration by inhalation, topical, transdermal, subcutaneous, intramuscular or
intravenous administration forms, rectal administration forms, and implants. For topical
application, the compounds according to the invention can be used in creams, gels,
ointments or lotions.
The injectable administration forms are particularly advantageous, conventionally
comprising the active compound placed in solution in water for injection, in the presence
of sodium chloride. The unit dose of active compound should be suitable for the desired
therapeutic effect; it may, for example, be between 0.1 and 100 mg of active ingredient.
According to another of its aspects, the present invention also relates to a method
for treating the pathological conditions indicated above, which comprises the
administration to a patient of an effective dose of a compound according to the invention
or a pharmaceutically acceptable salt thereof.
CLAIMS
1. Oligosaccharide compounds corresponding to formula (I):
in which:
- the wavy line denotes a bond located either below or above the plane of the
pyranose ring of the saccharide unit,
- R represents an -O-alkyI group, in which said alkyl group contains from 1 to 16
carbon atoms and is optionally substituted with one or more groups, which may be
identical or different, chosen from aryl and cycloalkyl groups,
- R2 represents a hydroxyl group or an -O-alkyI group,
- R3, R5, R6, R7 and R8, which may be identical to or different from one another,
represent either an -OS0 3 group or a hydroxyl group,
- R4 represents either an -NH-CO-alkyl group or an -O-alkyI group,
- R represents an -O-alkyI group, and
- n and m, which may be identical to or different from one another, represent
integers equal to 0 or 1,
in acid form or in the form of any one of the pharmaceutically acceptable salts
thereof.
2. Compounds according to Claim 1, in which n = 1 and m = 0 or else n = 0 and
m = 1.
3. Compounds according to Claim 1 or Claim 2, in which R represents an -O-alkyI
group, where said alkyl group contains from 1 to 8 carbon atoms and is optionally
substituted with 1 or 2 groups, which may be identical or different, chosen from aryl
groups.
4. Compounds according to any one of the preceding claims, in which R
represents an -O-methyl or -O-pentyl group and is optionally substituted with 1 or 2 phenyl
groups.
5. Compounds according to any one of the preceding claims, in which R3, R5, R6,
R7 and R8, which may be identical to or different from one another, represent either
an -OSO 3 group or a hydroxyl group, on the condition that at least one group among R3,
R5, R6, R7 and R8 represents an -OSO3 group.
6. Compounds according to any one of the preceding claims, in which R3, R5, R6,
R7 and R8 all represent -OSO3 groups.
7. Compounds according to any one of Claims 1 to 5, in which at least one of the
groups R3, R5, R6, R7 and R8 represents an -OS0 3 group and at least one of the groups
R3, R5, R6, R7 and R8 represents a hydroxyl group.
8. Compounds according to any one of Claims 1 to 5 and 7, in which R3, R5 and R6
represent -OS0 3 groups and R7 and R8 represent hydroxyl groups.
9. Compounds according to any one of the preceding claims, in which R4
represents an -NH-CO-alkyl group, where said alkyl group comprises from 1 to 4 carbon
atoms.
10. Compounds according to any one of the preceding claims, in which R
represents a methoxy group.
11. Compounds according to any one of the preceding claims, chosen from the
following compounds:
- methyl (sodium 3-0-methyl-2-0-sodium sulphonato-a -L-idopyranosyluronate)-
( 1 4)-(2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a -D-glucopyranosyl)-
( 1 4)-[(sodium 3-0-methyl-2-0-sodium sulphonato-a -L-idopyranosyluronate)-(1 4)-(2-
acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a -D-glucopyranosyl)-(1 4)]2-
(sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-(1 4)-2-
acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a-D-glucopyranoside (No. 1);
- pentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-
( 1 4)-(2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a-D-glucopyranosyl)-
( 1 4)-[(sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-(1 4)-(2-
acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a-D-glucopyranosyl)-(1 4)] 2-
(sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-(1 4)-2-
acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-p-D-glucopyranoside (No. 2);
- pentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-
( 1 4)-(2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a-D-glucopyranosyl)-(
1 4)-[(sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-
( 1 4)-(2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a-D-glucopyranosyl)-(
1 4)] 2-(sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-
( 1 4)-2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium sulphonato-p-D-glucopyranoside
(No. 3);
- 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1 4)-(2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a-
D-glucopyranosyl)-(1 4)-[(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1 4)-(2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-a-
D-glucopyranosyl)-(1 4)] 2-(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1 4)-2-acetamido-2-deoxy-3-0-methyl-6-0-sodium sulphonato-b-
D-glucopyranoside (No. 4);
- 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1®4)-(2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-a-D-glucopyranosyl)-(1 4)-[(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1®4)-(2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-a-D-glucopyranosyl)-(1 4)] 2-(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1 4)-2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-p-D-glucopyranoside (No. 5);
- 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1 4)-(2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-a-D-glucopyranosyl)-(1 4)-(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1®4)-(2-butanoylamino-2-deoxy-3-0-methyl-6-0-sodium
sulphonato-a-D-glucopyranosyl)-(1 4)-(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1®4)-(2-butanoylamino-2-deoxy-3-0-methyl-a -D-glucopyranoside)-
( 1 4)-(sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-(1 4)-2-
(butanoylamino)-2-deoxy-3-0-methyl-p -D-glucopyranoside (No. 6);
- 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1®4)-(2-[(3-methylbutanoyl)amino]-2-deoxy-3-0-methyl-6-0-
sodium sulphonato-a-D-glucopyranosyl)-(1 4)-(sodium 3-0-methyl-2-0-sodium
sulphonato-a-L-idopyranosyluronate)-(1®-4)-(2-[(3-methylbutanoyl)amino]-2-deoxy-3-0-
methyl-6-O-sodium sulphonato-a-D-glucopyranosyl)-(1 4)-(sodium 3-0-methyl-2-0-
sodium sulphonato-a-L-idopyranosyluronate)-(1®4)-(2-[(3-methylbutanoyl)amino]-2-
deoxy-3-0-methyl-a -D-glucopyranoside)-(1 4)-(sodium 3-0-methyl-2-0-sodium
sulphonato-a-L-idopyranosyluronate)-(1®-4)-2-[(3-methylbutanoyl)amino]-2-deoxy-3-0-
methyl-p-D-glucopyranoside (No. 7);
- 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1 4)-(2-0-butyl-3-0-methyl-6-0-sodium sulphonato-a-D-glucopyranosyl)-(
1 4)-(sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-
( 1 4)-(2-0-butyl-3-0-methyl-6-0-sodium sulphonato-a-D-glucopyranosyl)-(1 4)-(sodium
3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-(1 4)-(2-0-butyl-3-0-
methyl-a-D-glucopyranosyl)-(1 4)-(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1®-4)-2-0-butyl-3-0-methyl-p -D-glucopyranoside (No. 8); and
- 5-phenylpentyl (sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1 4)-(2-0-butyl-3-0-methyl-6-0-sodium sulphonato-a-D-glucopyranosyl)-(
1 4)-(sodium 3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-
( 1 4)-(2-0-butyl-3-0-methyl-6-0-sodium sulphonato-a-D-glucopyranosyl)-(1 4)-(sodium
3-0-methyl-2-0-sodium sulphonato-a-L-idopyranosyluronate)-(1 4)-(2-0-butyl-3-0-
methyl-a-D-glucopyranosyl)-(1 4)-(sodium 3-0-methyl-2-0-sodium sulphonato-a-Lidopyranosyluronate)-(
1 4)-2-0-butyl-3-0-methyl-a -D-glucopyranoside (No. 9).
12. Medicament, characterized in that it comprises a compound according to any
one of Claims 1 to 11, or a pharmaceutically acceptable salt thereof.
13. Pharmaceutical composition, characterized in that it comprises a compound
according to any one of Claims 1 to 11, or a pharmaceutically acceptable salt thereof, and
also at least one pharmaceutically acceptable excipient.
14. Compound according to any one of Claims 1 to 11, for use thereof in the
treatment of pathological conditions requiring activation of FGF receptors.
15. Compound according to Claim 11, for use thereof in the treatment of ischaemia,
such as cardiac ischaemia or lower limb ischaemia, the treatment of diseases associated
with narrowing or obstruction of the arteries or arteritis, the treatment of angina pectoris,
the treatment of thromboangiitis obliterans, the treatment of atherosclerosis, cicatrisation,
the treatment of post-angioplasty or post-endarterectomy restenosis; the treatment of
chronic ulcer and refractory ulcer in diabetic or nondiabetic patients, chronic or nonchronic
perforations of the eardrum, periodontitis, muscle regeneration and myoblast survival,
peripheral neuropathy, post-operative nerve damage, nerve deficiencies such as
Parkinson's disease, Alzheimer's disease, prion disease and neuronal degeneration in
alcoholics, dementia, bioartificial pancreas graft survival in diabetic patients, retinal
degeneration, stromal keratitis, pigmentary retinitis, osteoarthritis, pre-eclampsia, vascular
lesions and acute respiratory distress syndrome, post-traumatic cartilage and bone repair,
the repair and protection of hair follicles, and the protection and regulation of hair growth.
16. Compound of formula (II), in which Alk represents an alkyl group and Pg and
Pg' represent protecting groups:
17. Compound according to Claim 16, in which the Alk groups represent methyl
groups and Pg and Pg' represent, respectively, acetyl and benzyloxycarbonyl groups.
18. Compounds of formula (III), in which Alk represents an alkyl group, R is as
defined in any one of Claims 1 to 11, A represents an -NH-Pg" or -O-butyl group, and Pg,
Pg' and Pg", which may be identical to or different from one another, represent protecting
groups:
19. Compounds according to Claim 18, in which the Alk groups represent methyl
groups, R represents an -O-pentyl or -O-pentylphenyl group, Pg represents an acetyl or
benzoyl group, Pg' represents an acetyl or ie/f-butyldiphenylsilyl group, and A represents
an - NH-benzyloxycarbonyl or -O-butyl group.
20. Compounds of formula (IV), in which Alk represents an alkyl group, B
represents an azide or -O-alkyl group, Pg, Pg' and Pg", which may be identical to or
different from one another, represent protecting groups, and D represents an activating
group or an -O-acetyl group:
2 1. Compounds according to Claim 20, with the exception of the compound of
formula (IV) in which Alk represents a methyl group, B represents an azide group, Pg
represents a levulinyl group, Pg' and Pg" represent acetyl groups, and D represents a
trichloroacetimidate group.
22. Compounds according to either one of Claims 20 and 21, in which B
represents an -O-alkyl group.
23. Compounds according to any one of Claims 20 to 22, in which the Alk groups
represent methyl groups, B represents an -O-butyl group, Pg represents a benzyl or
levulinyl group, Pg' represents an acetyl or benzoyl group, Pg" represents an acetyl or
fe/t-butyldiphenylsilyl group and D represents a trichloroacetimidate or -O-acetyl group.
24. Compounds according to any one of Claims 20 to 22, in which the Alk groups
represent methyl groups, B represents an azide group, Pg represents a benzyl or levulinyl
group, Pg' represents an acetyl or benzoyl group, Pg" represents an acetyl or tertbutyldiphenylsilyl
group and D represents a trichloroacetimidate or -O-acetyl group, with
the exception of the compound of formula (IV) in which Alk represents a methyl group, B
represents an azide group, Pg represents a levulinyl group, Pg' and Pg" represent acetyl
groups, and D represents a trichloroacetimidate group.