CA2388663A1 - Use of quinic, shikimic acids and their derivatives for preparing mannose receptor ligands - Google Patents

Abstract

The present invention relates to the use of quinic, shikimic acids and their derivatives for the preparation of mannose receptor ligands, to the ligands thus prepared, as well as to the applications of these ligands, in particular for the preparation of medicaments and , more particularly for the preparation of vaccines and drugs intended to ensure the vectorization of an active principle towards target cells expressing the manno receptor or a receptor related thereto, the transfection of DNA and RNA sequences and its multiple uses in gene therapy, and the preparation of laboratory reagents, in particular diagnostic reagents.

Description

USE OF QUINIC, SHIHIMIC ACIDS AND THEIR
DERIVATIVES FOR THE PREPARATION OF LIGANDS OF THE RECEPTOR OF
MANNOSE
The present invention relates to (use of quinic acids, shikimic and their derivatives for the preparation of receptor ligands mannose, . to the ligands thus prepared, as well as to the therapeutic applications and diagnoses of these ligands.
Dendritic cells, which represent a particular line of leukocytes originating in the bone marrow and which are present in practically all the tissues of the organism, play a key role in the control of answers immune. Indeed, these cells appear to be the only cells capable in vivo to present antigens to naive and mature T cells, and to induce, by activation of these T cells, a T-helper immune response (J. BLANCHEREAU and RM STEINMANN, Nature, 1998, 392, 245-252).
Activation of T lymphocytes by ~ dendritic cells involves prior recognition and internalization of antigens by these past. However, it has been shown that the internalization of antigens by cells dendritics occurs not only by macropinocytosis, but also by a endocytosis mechanism mediated by a specific receptor for hydrates of carbon and called the mannose receptor (A. AVR.AMEAS et al., Eur. J.
Immunol., 1996, 26, 394-400).
The mannose receptor belongs to the family of C-lectins which represents a group of calcium-dependent glycoproteins forming connections non-covalent with carbohydrates. If, as its name (indicates, the mannose receptor binds preferentially to D-mannose, it appears bind also to other carbohydrates like L-fucose, D-glucose and N-acetyl-D-glucosamine which, like D-mannose, has two functions hydroxyls carried by two adjacent carbon atoms of the ring which constitutes. he seems to be the link between the mannose receptor and these carbohydrates established by the coordination of a calcium ion between amino acid residues of the site of recognition of this receptor and the two adjacent hydroxyl functions said carbohydrates (WI WEIS et al., Nature, 1992, 360, 127-134).

WO 01/09173

2 PCT / FR00 / 02194 Furthermore, as is the case for all lectins, the carbohydrate recognition by the mannose receptor, although specific, is weak, of the order of 10 '~ to 10'3 M. The simultaneous connection of many carbohydrate molecules at multiple recognition sites of the receiver mannose compensates for this low affinity. This phenomenon is called "
effect of group ".
It has also been shown, not only that internalization, by dendritic cells, mannosylated antigens is significantly greater than that of non-mannosylated antigens, but that a culture of clonal populations of T cells of specificity restricted to certain peptide-HLA complexes of class II, performed in the presence, on the one hand, of dendritic cells and, on the other hand, protein or mannosylated peptides results in stimulation of these lymphocytes 200 to 10,000 times higher than that obtained when these same proteins and peptides do are not mannosylated (MCAA TAN et al., Eur. J. Immunol., 1997, 27 2426-2435).
It therefore appeared possible to induce, stimulate or modulate an immune response by promoting the internalization of an antigen by the dendritic cells via the mannose receptor, by coupling this antigen with a chemical structure carrying several molecules of a carbohydrate specifically recognized by this receptor (MCAA TAN et al., ibid).
This is of particular interest in vaccinology and especially for the development of vaccines based on synthetic peptides. In effect, if the latter have (advantage of being both chemically defined, of precise composition, freed from the difficulties of organic production, purified and, finally, of easy distribution because of their stability, they present usually the major drawback of being weakly immunogenic.
This is how the inventors have proposed to modify antigens.
peptides by grafting them onto a dendrimeric structure comprising a heart of L-lysine tree type linked to several glycosyl residues and, in particular, to remains D-mannose (C. GR.ANDJEAN et al., Lleme Réunion Peptides et Proteins, 17-22 January 1999, AUSSOIS, FRANCE).
However, it appears that, from the perspective of development at a industrial scale of a drug and, in particular, of a vaccine, (use WO 01/09173

3 PCT / FR00 / 02194 natural carbohydrates like D-mannose or D-glucose, is susceptible to pose a certain number of difficulties, both technically and financial, in because these compounds are difficult to link to other structures chemical and are unstable in many reaction media - in particular, in the environments acids - and in many biological media.
The problem arises, therefore, of having compounds exhibiting, with respect to the receptor mannose, an affinity and a specificity as they can be recognized by this receptor and selectively bind to it, while being devoid of the disadvantages of natural carbohydrates.
However, the Inventors, always in the context of their research on the mannose receptor, found that, surprisingly, the acids quinique, shikimic and their derivatives which by their nature represent carbocyclic, chemically more stable compounds than natural carbohydrates, and therefore easier to use in synthesis processes, are able to mimic the D-mannose vis-à-vis its receptor, i.e. to selectively bind to this last, and therefore advantageously replace the hydrates of carbon natural in the preparation of ligands intended to promote (internalization of a substance by cells expressing this receptor or a receptor for the family of C-lectins being related to it.
The present invention therefore relates to the use of at least one compound corresponding to the general formula (Ia) and / or at least one compound respondent to the general formula (Ib) O Rs O R5 Rt0 4 OR3 RIO 4 OR3 (Ia) in which - Ri, Rz, R3 and R4 represent, independently of each other, an atom WO 01/09173

4 PCT / FR00 / 02194 hydrogen or a protecting group, - RS represents a group -OH, a group -SH, a group -NH-NH2, an atom halogen, or a linear, branched or cyclic, saturated or unsaturated, comprising from 1 to 18 carbon atoms and possibly from 1 to 12 atoms chosen from oxygen, sulfur and nitrogen, said carbon chain being optionally substituted with 1 to 16 halogen atoms and bearing eventually at least one nucleophilic group or at least one electrophilic group, as a mannose receptor ligand or any family receptor of C-lectins related to the mannose receptor, or for the preparation of such ligand.
For the purposes of the present invention, the term “group”
"Protecting" a protecting group for a hydroxyl function or a diol (in which case two groups chosen from R1, R2, R3 and R4 can be linked in a way covalent one to the other to form a cycle together), as defined in the book Protective Groups in Organic Synthesis, TW GREENE and PGM WUTS, Second Edition 1991, J. WILEY and Sons. By way of examples and without limitation, we can quote trimethylsilyl, triethylsilyl, trüsopropylsilyl groups, tertiobutyldimethylsilyl, tertiobutyldiphenylsilyl, trimethylsilylethylether, tert-butoxymethyl, methoxy-methyl, benzyloxymethyl, 2-methoxyethoxymethyl, methylthiomethyl, acetyl, or 2 ', 3'-dimethoxybutane-2', 3'-diyl.
Furthermore, the term “nucleophilic group u” is understood to mean a group having a pair of free electrons, i.e. a donor group electrons, and by group o electrophilic o a group with a molecular orbital vacant from low energy capable of interacting with the pair of free electrons of a group nucleophile. The presence of at least one nucleophilic or electrophilic group in the compounds of formulas (Ia) and (Ib) advantageously allows their incorporation in other structures, such as load-bearing structures, by reacting what group nucleophile or electrophile with corresponding reactive functions worn by said structure, as will be described in detail below.
In the context of the present invention, the nucleophilic group is, preferably chosen from amine groups, thiols, ~ 3-aminothiols, alcohols, hydrazide and hydrazide derivatives, hydrazine and hydrazine derivatives, hydroxylamine and derivatives of hydroxylamine, while the electrophilic group is preferably chosen from WO 01/09173

5 PCT / FR00 / 02194 a-oxo-aldehyde groups, -CORa (with for example Ra = H, halogen, imidazolyle, aromatic heterocycle), -COzRa (with for example Ra = H, aryl, succinimidyle, sulfosuccinimidyle, aromatic heterocycle) and its derivatives (anhydrides symmetrical, mixed anhydrides, activated esters), -OCORa (with for example Ra = halogen, imidazolyl, aromatic heterocycle), -OCOZRa (with for example Ra = aryl, heteroaryl, succinimidyl, sulfosuccinimidyl), -OCSRa (with for example Ra =
halogen, imidazolyl, aromatic heterocycle), -OCSORa (with for example Ra =
aryl, succinimidyl, sulfosuccinimidyl, aromatic heterocycle), -NRaCORb (with for example Ra = methyl and Rb = halogen, imidazolyl, heterocycle aromatic), -NRaCOORb (with for example Ra - H and Rb - aryl, succinimidyle, sulfosuccinimidyle), -NRaCSRb (with for example Ra = H and Rb = halogen, imidazolyl, aromatic heterocycle), -NR ~ CSORb (with for example Ra = H and Rb =
aryl, succinimidyl, sulfosuccinimidyl), -NCO, -NCS, maleimide and halides alkyls.
By way of examples and without limitation, in the formulas general (Ia) and (Ib) - suitable halogen atoms are bromine, chlorine and fluorine; and - suitable carbon chains are alkyl groups (methyl, ethyl, propyl, isopropyl, tert-butyl, pentyl, ...), alkenyl groups (vinyl, allyl, butenyl, pentenyl, ...), alkynyl groups (ethynyl, propynyl, butynyl, pentynyl, ...), cycloalkyl groups (cyclopentyl, cyclohexyl, ...), heterocycles (piperazine, ...), aryl groups (phenyl, cresyl, ...), groups heteroaryls (pyridine, pyrimidine, pyrazine, ...), groups polyethylene glycols or still polyamines.
The compounds corresponding to the general formulas (Ia) and (Ib) are mimics of D-mannose. They can be used, as such as D-mannose receptor ligands, or be linked to other compounds to drive to (obtaining ligands in which they will play the role of mimics of D-mannose.
Also, if it is appropriate that, in ligands such as ready to be used, the composed of general formulas (Ia) and (Ib) have at least two hydroxyl functions free vicinal, so as to be able to mimic D-mannose and allow these WO 01/09173

6 PCT / FR00 / 02194 ligands to be recognized by the mannose receptor and to bind to it last it is however, these compounds need not initially have two functions free vicinal hydroxyls.
Thus, in accordance with (Invention, it is entirely possible to use, for the preparation of a ligand, at least one compound of formula (Ia) and / or at least one compound of formula (Ib) comprising, on two carbon atoms adjacent to the ring, two hydroxyl groups protected by a protective group, the other groups carried by the ring atoms which may possibly be them also protected. This will even be desirable in a certain number of cases, in particular for avoid groups carried by the carbon atoms of the compound's cycle general formula (Ia) or (Ib) do not interfere during the coupling of said compound for lead to intramolecular cyclizations or dimerizations. Well heard, the groups protecting the two hydroxyl functions carried by the two atoms of adjacent carbon in the cycle will be removed prior to use ligand, elimination of other protecting groups being optional.
According to a preferred arrangement of (Invention, the compounds of general formulas (Ia) and / or (Ib) are chosen so that the groups -ORZ and -OR3 are in trans relation, so as to mimic the two carried hydroxyls by Ie D-mannose cycle that appear to be involved in the binding between this compound and its receiver.
Preferably, the compounds of formula (Ia) and / or (Ib) are such that RS represents a hydroxyl group (in which case the ring carries, in position 1, a group carboxyl).
Indeed, the presence of a carboxyl group on the cycle of compounds of formulas (Ia) and (Ib) offer the advantage of giving these compounds of numerous coupling possibilities, for example by (via a binding of amide, ester, thioester, hydrazide or hydroxamate type. Furthermore, these couplings can be easily achieved by simply activating this group carboxyl according to activation techniques conventionally used for synthesis peptide and, in particular, for peptide synthesis on solid support. These techniques are described, for example, in The Practice of Peptides Synthesis, M.
and A.
BODANSKY, 1984, SPRINGLER-VERLAG, Berlin.

WO 01/09173

7 PCT / FR00 / 02194 According to a particularly preferred provision of (Invention, the compound of formula (Ib) is such that R ~, RZ and R3 represent atoms hydrogen and RS represents a hydroxyl group (-OH). If, in addition, the groups -ORS and -OR2 se are in cis relationship, and the groups -ORZ and -OR3 are in relation trans, the configuration being 3R, 4S and SR, the resulting compound consists of (acid (-) - shikimique (or 3,4,5-trihydroxy-1-cyclohexene-1-carboxylic acid).
According to another preferred arrangement of (Invention, the compound of formula (Ia) is such that R ,, R2, R3 and R4 represent hydrogen atoms and RS
represents a hydroxyl group (-OH). If, in addition, the groups -ORZ and -OR3 is are in trans relation, while the -ORl and -OR4 groups are each in cis relation to the group -OR2, and the configuration is 1s ", 3R, 4s"
and SR, then the resulting compound consists of ((-) - quinic acid (or 1,3,4,5- acid tetrahydroxy-cyclohexane-carboxylic).
(-) - shikimic and (-) - quinic acids are available commercially, for example from the companies ALDRICH or ACROS.
According to yet another preferred arrangement of (Invention, the compound of formula (Ia) is such that R ,, R2, R3 and R4 represent atoms of hydrogen and RS represents a group -NH- (CHZ) 2-SH. It is then a derived from quinic acid.
In general, it is understood that the formulas (Ia) and (Ib), which represent quinic and shikimic acids and some of their derivatives, obtained by substitution of the hydroxyls carried in positions 2, 3 and 4 (and possibly in position 1) of the cycle, and by substitution of the function carbonyl located in position 1 of the cycle (RS group), are not limiting, and that other derived from quinic and shikimic acids could be used without leaving the frame of the presents Invention.
The present invention also relates to a compound useful in as a ligand for the mannose receptor or any receptor of the family of C-lectins related to the mannose receptor, or as a compound intermediate for the preparation of such a ligand, which compound being characterized in that he responds to the general formula (III) below WO 01/09173

8 PCT / FR00 / 02194 (BiM) rnW) nA ~ P ~ y (IU) in which - B 'meets one or more general formulas (IIa) or (IIb) below O W-OH

OH OH
(IIa) (IIb) where W represents a bond or a linear carbon chain, branched or cyclic, saturated or unsaturated, comprising from 1 to 18 atoms of carbon and optionally from 1 to 12 atoms chosen from oxygen, sulfur and nitrogen, said carbon chain being optionally substituted with 1 to 16 halogen atoms, - X represents the remainder of an oligopeptide X 'comprising from 1 to 6 acids amines, A represents the remainder of an at least difunctional compound A ′, - Y represents the remainder of a compound of interest Y ', - m is an integer between 1 and 32, 1 S - n is an integer between 0 and 32, - y is an integer between 1 and 4, and - M, N and P each represent a linking group, respectively between B1 and A
when n = 0 or B ~ and X when n is different from 0, between X and A when n is different from 0, and between A and Y, and include, independently of each other, a function chosen from oxime, hydrazone, amide, ester, thioester, hydrazide, hydroxamate, ether, thioether, amine, carbonate, cardamaie, thiocarbonate, thiocarbamate, urea, thiourea and thiazolidine.
In formula (III), as well as in all that follows, examples suitable halogen atoms and carbon chains are as described previously related to the compounds of general formulas (Ia) and (Ib).

WO 01/09173

9 PCT / FR00 / 02194 The compound corresponding to the general formula (III) therefore comprises * as compulsory elements ~ one or more B1 remains each responding to one of the general formulas (IIa) or (IIb), this or these remains being intended to play the role of mimes of the D-mannose vis-à-vis the mannose receptor and resulting from the bond between one or more compounds corresponding to the general formula (Ia) and / or (Ib) previously defined and the compound A ', optionally via X', and the deprotection of the hydroxyl groups, in the case where the said compound or compounds of general formula (Ia) and / or (Ib) initially has groups protected hydroxyls;
~ one or more residues Y of a compound Y 'corresponding to a compound of interest;
the remainder A of a compound A ′ having at least a double functionality, since its role is to carry both the B1 residue (s) and the remains Y;
and * as accessory element (s), the X residue (s) of a compound X ', this or these X remnants playing the role, when present, of spacer arms between BI
and A, from so as to control the distance between the different remains B1, when many of them are present in the compound of general formula (III) (i.e.
say when m is greater than 1). Although X residues of oligopeptide nature have been described, it would not go beyond the scope of the present invention to using, as compound X ', compounds of a different nature, provided they be able to play the role of spacer arm between B1 and A. As examples and but not limited to, there may be mentioned mercaptopropionic acid, thioamines, bromoamines and polyethylene glycol type spacer arms such as 4,7,10-trioxa-1,13-tridecane diamine.
For the purposes of the present invention, the term “compound”
of interest u a compound which one seeks to promote finalization by cells via the mannose receptor or a related C-lectin family receptor at this last (dendritic cells, cells of monocyte-macrophage lines, ...).
To this end, the compound of interest can be linked to the remainder A, at the rate of 1 to 4 molecules of compound of interest per molecule of compound of general formula WO 01/09173

10 PCT / FR00 / 02194 (III), by linking groups P each resulting from the reaction between two groups functional carried, for the first, by said compound of interest and for the second, by compound A '. A suitable ligand is obtained, by the presence of the residue (s) B ~, at specifically bind to the mannose receptor or any receptor related to it latest and, by its link with the compound of interest, to promote internalization of the last by cells expressing this type of receptor.
According to an advantageous arrangement of the invention, the compound A 'is formed of a chain comprising several remains of at least one compound trifunctional, linked to each other by covalent bonds, and clean to offer, to level of the free functional groups of these remains (i.e. groups which are not engaged in a covalent bond), several Site (s anchor for the compound (s) of general formula (Ia) and / or (Ib) or, when X
is present, compound X ', and for compound Y'.
According to a preferred form of this arrangement, the compound A ' includes from 2 to 32 and preferably 4 to 16 residues of a compound trifunctional.
In accordance with the invention, the compound A ′ can be presented either in a linear form, either in a branched form and, in particular, of the type tree-like (called "dendrimeric"). Examples of branched structures particularly well suited to the preparation of a receptor ligand for mannose according to (Invention are illustrated in Figures 1A and 1B which show, from way very schematic, a compound A 'in the form of two different dendrimers.
Preferably, the compound A ′ comprises a chain of acids amines, identical or different, selected from the group consisting of lysine, fhydroxylysine, serine, threonine, cysteine, fornithine, acid aspartic and (glutamic acid. This amino acid can be both of the L series and of D series and compound A 'can even include both L-series residues and of residues of series D.
A compound A 'based on lysine is preferred, (use of lysine being in fact particularly well suited to the production of a ligand of the receiver mannose according to (Invention, insofar as it has been shown that macromolecules containing lysine are devoid of intrinsic immunogenicity (TAM, Proc. Natl.
Acad Sci. USA, 1988, 85, 540).

WO 01/09173

11 PCT / FR00 / 02194 However, it is possible to use, as compound A ′, sequences of trifunctional compounds other than the amino acids mentioned this-above, provided that they have satisfactory biocompatibility. AT
title examples and in a nonlimiting manner, we can cite the 3,3'-iminobis (propylamine), the 2,2'-iminobis (ethylamine), gallic acid, tris (2-carboxyethyl) nitromethane (M.
BRETTEICH et al., Synlett, 1998, 1396), tris (hydroxymethyl) aminomethane (P.
R.
ASHTON et al., J. Org. Chem., 1998, 63, 3429) and (dihydroxy) propylamine.
Incidentally, the compound A ′ may also comprise one or more several residues corresponding to one or more different compounds of the composed at less trifunctional which constitutes its basic element. This or these remains don't have principle not intended to ensure any connection between the different elements composing the ligand. In fact, their presence is either intended to confer on this ligand additional characters such as one or more additional possibilities of coupling (for example, with a view to the subsequent production of a dimer of this ligand), is linked to the need to use, during its preparation, compounds additional, such as spacer arms. It goes without saying that these compounds, all while being different from the at least trifunctional compound constituting (basic element of the compound A ′, will preferably be of the same nature as the latter. So by example, they will preferably be amino acids in the case where the compound A ' understands an amino acid.
In accordance with (Invention, n, which represents the number of remains X
present in the compound of general formula (III), can be ~ is equal to 0, in which case the residue (s) B 1 are directly linked otherwise A, . is between 1 and 32, in which case, if n is equal to 1, the one or more remainders B ~ are linked to remainder A by (through a single remainder X, while that if n is different from 1, each remainder X can serve as a connecting element between one or many remains B 'and the rest A.
As for the linking groups M, N and P, they are chosen, independently of each other, among the liaison groups that include a function chosen from oxime, hydrazone, amide, ester, thioester, hydrazide, hydroxamate, ether, thioether, amine, carbonate, carbamate, thiocarbonate, WO 01/09173

12 PCT / FR00 / 02194 thiocarbamate, urea, thiourea and thiazolidine. As examples and so no-limiting - an oxime bond can be formed by reaction of a group O-alkylamine with a carbonyl group, - a hydrazone bond can be formed by reaction of a group hydrazine with a carbonyl group, - an amide bond (respectively, ester) can be formed by reaction of an amine group (respectively, alcohol) with an acid group activated or acid anhydride or halide, - a thioester bond can be formed by reaction of a group thiol with an activated acid group or anhydride or acid halide, - a hydrazide bond can be formed by reaction of a group hydrazine with an activated acid group, anhydride or acid halide, - an ether bond can be formed by reaction of an alcohol group with an alkyl halide group, - a thioether bond can be formed by reaction of a thiol with an alkyl halide group, - an amine bond can be formed by reaction between a group amine and an alkyl halide, - a carbonate bond can be formed by reaction of a group chloroformate with an alcohol group, - a carbamate bond can be formed by reaction of a group chloroformate with an amine group, - a thiocarbonate bond can be formed by reaction of a chlorothioformate group with an alcohol group, - a thiocarbamate bond can be formed by reaction of a chlorothioformate group with an amine group, - a urea bond can be formed by reaction of a group isocyanate with an amine group, - a thiourea bond can be formed by reaction of a group isothiocyanate with an amine group, WO 01/09173

13 PCT / FR00 / 02194 - a thiazolidine bond can be formed by reaction of a group (3-aminothiol with a carbonyl group, while - A hydroxamate bond can be formed by reaction between a activated acid group and a hydroxylamine group.
It will therefore be understood that, in the general formula (III), B ', A and X represent the remains of compounds naturally bearing groups functional suitable to lead, by reaction on each other, to obtaining groups of binding M, N and P as defined above, or of compounds having been modified from so as to carry such functional groups.
In this regard, it should be noted that, in accordance with the invention, the compound corresponding to the general formula (III) can be prepared - either by a convergent synthesis, that is to say by assembling commercially available and possibly modified compounds for needs of the preparation of the compound corresponding to the general formula (III), and / or of compounds previously synthesized.
Thus, for example, the compound of general formula (III) can be obtained by first coupling the compound (s) meeting the formula general (Ia) and / or (Ib) with compound X ', then coupling the compound resulting from this first coupling with compound A ', and finally by coupling the compound resulting from this second coupling with the compound Y ′, the latter coupling being eventually monitoring of the deprotection of the hydroxyl groups carried by the residue (s) B ~, in the case or the compound (s) of formula (Ia) and / or (Ib) carry such groups.
Alternatively, the compound of general formula (III) can be obtained by coupling a compound resulting from the coupling between one or more compounds corresponding to the general formula (Ia) and / or (Ib) and the compound X ', with a compound resulting from the coupling between compound A 'and compound Y', the latter coupling which can, again, be followed by deprotection as mentioned above.
Of course, any other assembly order can also be envisaged provided that it makes it possible to obtain a compound corresponding to the formula general (III).
- either by a recurring synthesis strategy, particularly in the case where the compounds Y ', A' and X 'consist of amino acids. So, the WO 01/09173

14 PCT / FR00 / 02194 compounds Y ', A' and X 'can be formed and joined together by addition successive amino acids that compose them, for example according to techniques of peptide synthesis on solid support derived from the method initially described by MERRIFIELD (The Chemistry of Solid Phase Peptide Synthesis, STEWART and YOLJNG Ed., 1984).
- or again by a partly convergent synthesis strategy and partly recurrent.
According to a preferred arrangement of (Invention, the compound meets the general formula (III) in which m is an integer between 4 and 16, n is an integer between 2 and 8, X represents the remainder of a oligopeptide comprising 2 to 4 amino acid residues, while A represents the remainder of a sequence comprising from 2 to 8 lysine residues and occurring under the form of a dendrimer.
According to another preferred arrangement of the invention, the compound of general formula (III) is such that B 'represents the remainder of one or more compounds chosen from ((-) - shikimic acid and (-) - quinic acid.
According to (use for which the ligand is intended, the compound of interest Y ' can be an antigen and, in particular, a vaccine antigen such as a peptide of synthesis corresponding to one or more epitopes of a protein of an agent pathogen, a protein or polysaccharide subunit antigen, a fraction purified bacterial or viral, an antibody such as an immunoglobulin or a serum antilymphocytes, a nucleic acid or a fragment of a nucleic acid (sequence RNA or DNA) coding for a protein of interest or for one or more antigenic determinants of this protein, a drug active ingredient such as immunostimulant, immunosuppressant, cytostatic or antibiotic, and can, at as such, to be as well a substance of protein, peptide nature, acidic or polysaccharide, lipid, lipoprotein, lipopolyoside, polynucleotide, one compound from chemistry.
The compound of interest can also be a detectable marker suitable for revealing the presence of the ligand in the medium where it turns found, such as a radioisotope, a fluorochrome or an enzyme.
Of course, it is also possible that the compound of interest Y 'is WO 01/09173

15 PCT / FR00 / 02194 made up of an association of several substances of different nature such as through for example, an antigen or antibody linked to a detectable label.
The present invention also relates to a compound useful as as an intermediate compound for the preparation of a receptor ligand for mannose or any receptor of the C-lectin family related to the receptor of the mannose, which compound is characterized in that it corresponds to the general formula (V) above after (B2M) rn (XN) nA (V) in which - BZ responds to one or more general formulas (IVa) or (IVb) below O W_ O W_ s 2 2 s (Na) (IVb) where R ,, R2, R3, Rd and RS are as defined above in relationship with the compounds of general formulas (Ia) and (Ib), and where W is such that defined previously related to the compounds of general formulas (11a) and (IIb), - X represents the remainder of an oligopeptide X 'comprising from 1 to 6 acids amines, A represents the remainder of an at least difunctional compound A ′, - m is an integer between 1 and 32, - n is an integer between 0 and 32, and - M and N each represent a linking group, respectively between B2 and AT
when n = 0 or BZ and X when n is different from 0, and between X and A when n is different from 0, and include, independently of each other, a function chosen from the oxime, hydrazone, amide, ester, thioester, hydrazide, hydroxamate, ether, thioether, amine, carbonate, carbamate, thiocarbonate, WO 01/09173

16 PCT / FR00 / 02194 thiocarbamate, urea, thiourea and thiazolidine.
Thus, in the general formula (V), the residue (s) BZ result from the coupling of one or more compounds corresponding to the general formula (Ia) and / or (Ib) with compound A ', optionally via X'.
In the compound of general formula (V), M, N, m, n, as well as the compounds A 'and X', are as described in relation to the general formula (III).
Furthermore, the compound of general formula (V) can be, like the compound corresponding to the general formula (III), prepared by a synthesis convergent, by a recurrent synthesis, or by a synthesis for part convergent and partly recurring.
According to a preferred provision of the invention, the compound of general formula (V) is such that Bz represents a remainder of one or more compounds chosen from ((-) - shikimic acid and ((-) - quinic acid, possibly form protected.
The present invention also relates to a compound useful as as an intermediate compound for the preparation of a receptor ligand for mannose or any receptor of the C-lectin family related to the receptor of the mannose, which compound is characterized in that it corresponds to the general formula (VI) below (B ~ M) n, A (V1) in which - B1 and M have the same meaning as in the general formula (III), - m is an integer between 50 and 500, and - A represents the remainder of a polymer A 'capable of forming a non-complex covalent with a compound of interest, such as a nucleic acid.
Within the meaning of the present invention, the term “polymer u” means a set of molecules of variable molecular weight, resulting from the sequence of several monomers.
This polymer is chosen from compounds capable of giving rise, in contact with a compound of interest, in the formation of a non-covalent complex with this composed, for example by means of ionic bonds, of bonds dipolar WO 01/09173

17 PCT / FR00 / 02194 or electrostatic interactions, the complex thus obtained then being susceptible to be used as a ligand for the mannose receptor or any receptor of the family of C-lectins related to the latter.
Preferably, the polymer A 'is a cationic polymer. As examples and without limitation, suitable cationic polymers are lysine polymers such as those marketed by SIGMA under the references P0296, P6403, P4408, P7886, P0899, P 1149 or even P0124, and polyethyleneimines, such as that marketed by FERMENTAS under the reference EXGEN 500 as a cell transfection reagent.
The present invention also relates to a compound useful in as a ligand for the mannose receptor or any receptor of the family of C-lectins related to the mannose receptor, which compound is characterized in this that it is in the form of a non-covalent complex between a compound corresponding to the general formula (VI) as defined above and a compound of interest, such as a nucleic acid.
The ligands in accordance with the invention are of great interest. Indeed, while being able to be very satisfactorily recognized by the receiver mannose and to specifically bind with it they have the advantage of could be easily prepared and at costs compatible with an operation industrial, in particular by the numerous techniques of peptide synthesis on a support solid.
These ligands are likely to find many applications as - the preparation of medicines and, more particularly, the preparation of vaccines and drugs for vectorization of a active ingredient towards target cells expressing the mannose receptor or a related receptor, - the transfection of DNA and RNA sequences and its multiples uses in gene therapy; in the latter case, we will use ligands themselves in the form of non-covalent complexes between a compound of formula general (VI) and a polynucleotide sequence, and - the preparation of laboratory reagents and, in particular, of diagnostic reagents.

WO 01/09173 1 g PCT / FR00 / 02194 The present invention also relates to the use of at least one compound corresponding to the general formula (Ia) and / or at least one compound respondent to the general formula (Ib) for the preparation of a medicament capable of link to mannose receptor or any receptor of the C-lectin family related to S mannose receptor.
The present invention also relates to a compound useful as as a mannose receptor ligand or any receptor of the C- family lectins related to the mannose receptor, as previously defined, for (use in as a medicine.
Such a drug is likely to find in the first place applications in all therapeutic indications in which research induction, modulation or suppression of immune responses and, more particularly those that are under the control of T cells.
examples and in a nonlimiting manner, we can cite - prevention and treatment of infectious diseases and, in particular, vaccinology, in which case the compound of interest will be an antigen vaccine or a nucleic acid or a nucleic acid fragment encoding a protein vaccine or for one or more antigenic determinants of such protein, and non-specific anti-infectious immunotherapy;
- treatment of autoimmune diseases such as lupus disseminated erythematosus, rheumatoid arthritis or WEGENER disease;
- the treatment of congenital or acquired immune deficiencies;
- prevention or treatment of hypersensitivity states;
- the treatment of cancer pathologies; and - prevention of rejection in organ transplants.
This medication is also likely to be used for vectorization of a non-immunomodulatory active principle towards cells targets expressing a receptor of the C-lectin family. So, for example, compounds useful as ligands according to the invention can find applications in antibiotic therapy, for the vectorization of antibiotics such as fluoroquinones or colistin to cells of the lines of infected monocyte-macrophages by bacteria or by mycobacteria, or in the treatment of diseases related to a cellular enzyme deficiency such as GAUCHER disease, in which case they will be used to vectorize the deficient enzyme to the KUPFER cells.
The present invention also relates to a compound useful as as a mannose receptor ligand or any receptor of the C- family lectins related to the mannose receptor, as previously defined, for use in vitro.
The present invention further relates to a diagnostic reagent, characterized in that it comprises a compound useful as a ligand for receiver mannose or any receptor of the C-lectin family related to receiver mannose, as previously defined.
In addition to the foregoing, the present invention includes still other provisions which will emerge from the additional description which follows, who relates to examples of the preparation of compounds useful as compounds intermediates for the preparation of mannose receptor ligands conform to (Invention, mannose receptor ligands according to the invention and demonstration of the biological properties of these ligands, as well as in the drawings annexed wherein - Figures 1A and 1B illustrate two examples of conformations branched which may be presented by the compound A ';
- Figures 2A, 2B and 2C illustrate the chemical structures of six compounds useful as intermediates for the preparation of ligands in accordance with the invention;
- Figures 3 and 4 illustrate the chemical structures of 4 ligands according to the invention prepared from the compounds shown in the Figures 2A and 2B;
- Figure 5 illustrates the chemical structures of two ligands corresponding to formula (III) in accordance with the invention, called 33F and 34F;
- Figures 6 and 7 illustrate the chemical structures of four ligands corresponding to formula (III) in accordance with the invention, called 41F, 42F, 43F
and 44F;
- Figures 8 and 9 illustrate the chemical structures of a derivative of quinic acid (compound G), two dendrimers (compounds H and I) and two peptide antigens (compounds J and K) usable to form ligands respondent to formula (III) in accordance with the invention, called 51 and 52;
- Figures 10 and 11 illustrate the chemical structures of two ligands corresponding to formula (III) in accordance with the invention, called S1 and 52;
- Figure 12 illustrates the ability of methyl quinate to inhibit internalization of compound 31F, carrying residues of D-mannose, such as describes in Example 8 below;
- Figure 13 illustrates, in the form of curves, the values average fluorescence intensities presented by cells dendritics treated with 2 ligands in accordance with (Invention (- ~ - and - ~ -) and with 2 compounds witnesses (-o- and -a-) at concentrations between 1 and 10 ~ M;
- Figure 14 illustrates, in the form of curves, the values average fluorescence intensities presented by cells dendritics treated with 2 ligands in accordance with (Invention (- ~ - and - ~ -) and with 2 compounds witnesses (-o- and -o-) at a concentration of 5 p, M after pre-incubation of the said cells in presence of mannan solutions of concentrations between 10 '~ and 10 mg / ml;
- Figure 15 illustrates the percentage of specific internalization of di-, tetra- or octavalent ligands by the mannose receptor, as described in Example 8 below;
- Figure 16 illustrates the chemical structure of a ligand called 61F, consisting of a dendrimer comprising 4 L-lysine residues carrying 4 residues of D-mannose, as well as a fluorescein molecule (FITC), as described in Example 9 below;
- Figure 17 illustrates the percentage of positive cells detected when incubating Cos-1 cells with different compounds, such as described in Example 9 below.
It should be understood, however, that these examples are given for illustrative purposes only (subject of (Invention and do not constitute in no way a limitation.
In the following, we will use the following formulas Ac20: acetyl anhydride Boc: t-butyloxycarbonyl 'Bu: tert-butyl DCC: dicyclohexylcarbodümide DCM: dichloromethane DIEA: düsopropylethylamine DMF: dimethylfonnamide eq. : equivalent EDTA: ethylene tetraacetic diamide ESI-MS: electrospray mass spectrometry Fmoc: 9-fluorenylmethoxycarbonyl HOBt: N hydroxybenzotriazole HBTU: N [(1H benzotriazol-1-yl) hexafluorophosphate oxide (dimethylamino) -methylene] -N methylmethanaminium MBHA: 4-methylbenzhydrylamine 1 S Mtt: 4-methyltrityl NMP: N methylpyrrolidone PBS: phosphate buffer Pmc: 2,2,2,5,7,8-pentamethylchroman-6-sulfonyl NMR: nuclear magnetic resonance RP-HPLC: high performance liquid chromatography in reverse phase TFA: trifluoroacetic acid TNBS: 2,4,6-trinitrobenzene sulfonic acid TOF-PDMS: time of flight spectrometry and plasma desorption Trt: trityle Furthermore, in the context of the following examples ~ the optical rotations were measured with a PERKIN polarimeter ELMER 241 and the values [a.] P are expressed in units of 10- ~ degrees.cm2 / g;
~ high performance semi-reverse phase liquid chromatography preparatory and analytical work was carried out using a SHI1 ~~ IADZU system LC-9A or LC-4A, using a BECKMANN ultrapore C-8 column (300 A, S p, m, 4.6 x 250 mm) or HYPERSIL hyperprep C-18 (300 A, 8 pm, 15 x 500 mm), a flow rate of 1 or 3 ml / minute, detection at 215 or 230 nm, and fun of the 5 following elution systems - system A: 0.05% TFA in Skin - system B: 0.05% TFA in an acetonitrile / water mixture (80:20) - system C: 0.05% TFA in an acetonitrile / water mixture (60:40) - system D: 50 mM phosphate buffer, pH 6.95 - system E: a mixture of 50 mM phosphate buffer, pH 6.95, and acetonitrile (50:50) - system F: 0.05% TFA in a water / isopropanol mixture (60:40);
~ TOF-PDMS spectra were recorded using a spectrometer mass to plasma desorption APPLIED BIOSYSTEMS Bio-Ion 20;
~ ESI-MS spectra were recorded using (using a mass spectrometer at source Micromass Quatro II electrospray ionization; while ~ 1 H and 3 C NMR spectra were recorded using a spectrometer mass BRUKER DRX 300 or DRX 600; chemical shifts are expressed in ppm, taking as internal standard, tetramethylsilane, that of the salt of sodium of deuterated trimethylsilylpropionic acid.
EXAMPLE 1: PREPARATION OF USEFUL COMPOUNDS AS
INTERMEDIATE COMPOUNDS FOR THE PREPARATION OF LIGANDS
CONFORM TO THE INVENTION
Six compounds in the form of dendrimers and corresponding to the general formula (V) in which ~ m represents an integer equal to 4, 8 or 16, ~ BZ represents an acid (-) - shikimic or (-) - quinic residue, ~ n is an integer equal to 2, 4 or 8 (in the case where m is respectively equal to 4, 8 and 16), ~ X represents the remainder of a tripeptide of sequence (S1) below Lys-Cys (tert-butylthio) -Gly-OH (S1) ~ A represents the remainder of a dendrimer comprising 2, 4 or 8 (in the case where m East equal respectively to 4, 8 and 16) residues L-lysine and having, on the one hand, 2, 4 or 8 chloroacetyl groups for its binding with an equivalent number of tripeptides and, on the other hand, a free amine group capable of allowing its binding with a molecule of interest, ~ M represents an amide group (-CO-NH-), while N represents a group thioether (-CHZ-S-CHZ-CO-), have been prepared.
The preparation of these compounds was carried out using a convergent synthesis strategy, i.e. by proceeding - firstly, the preparation of the dendrimers A ', - then, to the preparation of the tripeptides of sequence (S 1) and to their coupling with two (-) - shikimic or (-) - quinic acid molecules, and - finally, to the coupling of the dendrimers A 'and of the tripeptides carrying the residues acid (-) - shikimic or (-) - quinic, by deprotection of the thiol function of these tripeptides and by forming a thioether bond between this function thiol and a chloroacetyl group of said dendrimers A '.
The chemical structures of these compounds, which will be referred to as in the following ~ compounds 21, 23 and 25, for compounds comprising respectively 4, 8 and 16 acid (-) - shikimic, and ~ compounds 22, 24 and 26, for compounds containing respectively 4, 8 and 16 acid (-) - quinic residues, are shown in Figures 2A, ZB and 2C.
1.1 - Preparation of the dendrimers A ' Each dendrimer A 'was prepared on a scale of 0.5 mmol by synthesis on a solid support, from 15 g of an MBHA resin initially loaded at 0.4 mmol / g (SENN CHEMICALS company) and using ~ a Boc / benzyl strategy for the protection of the amino acids to be coupled, ~ a mixture of HBTU / HOBtIDIEA in DMF, as activating reagents for amino acid couplings, and ~ Ninhydrin tests - (KAISER et al., Anal. Biochem., 1970, 34, 595) and at TNBS - (HANCOCK and BATTERSBY, Anal. Biochem., 1976, 71, 261) for the control of coupling and deprotection operations.
In order to lower the charge in free amine functions of the resin to a value of 0.1 mmol / g, 0.25 equivalent (per amine function to react) of Boc- / 3-Ala-OH was initially coupled to this resin, in the presence of a HBTU / HOBt / DIEA mixture (0.25: 0.25: 0.75 eq. per amine function in front react) in DMF (reaction time: 1 hour). Then, the amino groups of the resin remained free were protected by acetylation, by reacting the resin with a Ac20 / DIEA / DCM mixture (5:10:85) for 10 minutes.
The second coupling - actually corresponds to the first coupling of an L-lysine - was produced using 4 equivalents (per amine function in front of react) of Boc-L-Lys (2-chlorobenzyloxycarbonyl) -OH, i.e. an L-lysine whose side chain is protected by a 2-chlorobenzyloxycarbonyl group, in presence of a HBTU / HOBt / DIEA mixture (4: 4: 12 eq. per amine function in front react) in DMF (reaction time: 45 minutes).
The third pairing was done using 4 equivalents of Boc-L-Lys (Boc) -OH, in the presence of a HBTU / HOBt / DIEA mixture (4: 4: 12 eq.
per amine function to react) in DMF (reaction time: 45 minutes).
At the end of this third coupling, a third of the resin was deprotected, then acylated with 8 equivalents of preformed chloroacetic anhydride (via a activation by DCC), to obtain a dendrimer (called compound 13 in this which follows) comprising 2 L-lysine residues and having on the one hand, a group free amine (which is none other than the s-amine group of the first L-lysine having been coupled with (3-alanine) and on the other hand, 2 chloroacetyl groups.
The other two thirds of the resin were coupling to obtain the attachment of two additional L-lysines. This coupling to was carried out with 4 equivalents of Boc-L-Lys (Boc) -OH, in the presence of a mixture HBTU / HOBt / DIEA (4: 4: 12 eq per amine function to react) in DMF
(reaction time: 45 minutes).
After the fourth coupling, half of the remaining resin (i.e.
1/3 of the initial resin quantity) was in turn unprotected and acylated by 8 equivalents of preformed chloroacetic anhydride to obtain a dendrimer (referred to as compound 14 below) comprising 4 Ll ~ sine residues and presenting to both a free amine group and 4 chloroacetyl groups.
The second half of the remaining resin was subjected to a fifth coupling with 4 equivalents of Boc-L-Lys (Boc) -OH, as described previously. Deprotection and acylation of the resin by chloroacetic anhydride were rolled over again to get a 8-residue dendrimer L-lysine (referred to as compound 15 below) presenting a free amine group and 8 chloroacetyl groups.
In practice, during each coupling operation, the HBTU (1 eq.
relative to the amino acid to be coupled), after dissolution in DMF, was added to a solution containing (amino acid to be coupled (0.25 eq. relative to functions -NHZ for ~ 3-alanine, and 4 eq. for the coupling of lysines), of HOBt (1 eq. through compared to the amino acid) and DIEA (3 eq. compared to the amino acid) in some DMF. The whole was mixed for 1 minute at room temperature, then added resin previously soaked in DMF. The suspension was stirred mechanically throughout the duration of the reaction.
In addition, between each coupling operation, the resin was successively filtered, washed 3 times with DMF (3 x 2 minutes) and DCM
(3 x 2 minutes), treated with a TFA / DCM mixture (50:50, 1 x 2 minutes, 1 x 20 minutes) suitable for allowing the cleavage of the Boc protective groups, washed again with DCM (2 x 2 minutes), neutralized with a DIEA / DCM mixture (5:95; 3 x 1 minute) and washed one last time with DCM (2 x 1 minute).
The cleavage of the dendrimers and their deprotection were obtained in reacting the resin with 11 ml of an HF / anisole mixture (10: 1) per gram of resin, at 0 ° C and for 60 minutes. As a result, they were successively precipitated in cold tert-butyl methyl ether, centrifuged, dissolved in Skin, lyophilized and purified by a RP-HPLC will be preparative [gradient (A / 8):
100: 0 to 50:50 in 120 minutes].
Were so obtained ~ 160 mg of dendrimer 13 (yield: 52%); TOF-PDMS spectrum: m / z 498 (M + H) +;
~ 215 mg of dendrimer 14 (yield: 42%); TOF-PDMS spectrum: m / z 928 (M + Na) +;
and ~ 182 mg of dendrimer 15 (yield: 20%); TOF-PDMS spectrum: m / z 1726 (M + H) +.

WO 01/09173 2 (PCT / FR00 / 02194 1.2 - Preparation of trineptides carrying the acid (-) - shikimiaue and (-) - guiniaue The preparation of tripeptides carrying acidic remains (-) - shikimique and (-) - quinique was carried out on solid support, in synthesizing in a first time the peptide of sequence (S1) and by coupling in a second time one molecule of this tripeptide with 2 molecules of (-) - shikimic acid or 2 molecules (-) - quinic acid derivatives, by forming a bond between amine groups N-terminal L-lysine of said tripeptide and the carboxyl group of said acids.
Were so obtained ~ on the one hand, N ", N 'di - [(3R, 4S, SR) -3,4,5-trihydroxy-1-cyclohexenecarbox-1-yl] -L-lysyl- [S- (tert-butylthio)] - L-cysteyl-glycine, which will be called compound 1 in this which follows, and ~ on the other hand, IVa, N ~ di - [(1s ~, 3R, 4s ", SR) -1,3,4,5-tetrahydroxy-1-cyclohexane carbox-1-yl] -L-lysyl- [S- (tert-butylthio)] - L-cysteyl-glycine, which will be called compound 3 in the following.
a) Preparation of compound 1 Compound 1 was prepared on a 2 mmol scale using (using a Fmoc-glycine p-benzyloxybenzyle resin (available under the name Fmoc-Gly-O-Wang with NOVABIOCHEM) responsible for 0.8 mmol / g, and using ~ an Fmoc strategy for the protection of the amino functions of amino acids at couple, ~ a mixture of HBTUIDIEA in NMP, as activating reagents for the amino acid couplings, ~ a piperidine / NMP mixture to remove the Fmoc protective groups, and ~ Ninhydrin and TNBS tests to control the reactions of coupling and deprotection.
After elimination of the Fmoc groups protecting the functions amine from the glycine residues of the resin, by treatment of the latter with a mix piperidine / NMP (20:80) for 20 minutes, coupling the residue S- (tert-butylthio) L-cysteine was produced using 2 equivalents (per amine group before react) from Fmoc-L-Cys (tert-butylthio) -OH in the presence of the HBTCJ / DIEA mixture (2: 3 eq.) In NMP (reaction time: 40 minutes). The use of a tert-butylthiol as a protective group for the thiol function of the L-cysteine residue has been chosen due to the fact that this group is both insensitive to conditions reactionary used during the coupling and cleavage operations, while being susceptible to be easily eliminated by the action of trialkylphosphines.
This first coupling was followed by a second coupling intended to fix tripeptide L-lysine, which was made using 2 equivalents of Fmoc-L-Lys (Fmoc) -OH in the presence of 5 equivalents of HBTU / DIEA mixture (2: 3 eq.) in NMP (reaction time: 40 minutes).
Then, the two acid molecules were coupled (-) - shikimic by activating 2 equivalents (per amine function to react) of this acid by, an HBTU / DIEA mixture (1: 1 eq.) in NMP for 30 seconds, and by adding said acid to the resin previously soaked in NMP (duration of the reaction: 40 minutes).
At the end of this coupling operation, the solvent was filtered, the resin was washed successively with NMP (3 x 2 minutes) and DCM (3 x 2 minutes), then dried.
The cleavage of compound 1 was carried out by treating the resin with 25 ml of a TFA / HZO / MezS mixture (95: 2.5: 2.5) at room temperature for 2 hours. The solution was concentrated under reduced pressure and the residue so got was dissolved in water and then subjected to lyophilization leading to (obtaining 1.64 g of a pale yellow powder. 107 mg of this powder has been purified by a RP-HPLC semi-preparative [gradient (AB): 100: 0 to 50:50 in 110 minutes], this who has allowed to obtain, after lyophilization, 15 mg of pure compound 1, i.e.
yield of 14%.
Compound 1 has the following characteristics Optical rotation: [a] D -117 (c 0.19, H20);
1 H NMR spectrum: 8H (300 MHz; HZO-D20 90:10) 1.16 (3 x 3 H, 3 s, C (CH3) 3), 1.14 1.35 (2 H, m, 2 lysyl yH), 1.37-1.44 (2 H, m, 2 lysyl SH), 1.62-1.75 (2 H, m, 2 lysyl (3-H), 2.00-2.11 (2 H, m, 2 6-H), 2.59 (1 H, dd, J 11.3 and J 17.4, 1 6-H), 2.60 (1 H, dd, J 11.1 and J 17.0, 1 6-H), 2.90 (1 H, dd, Ja, p 7.0 and Ja, a ~ 14.1, 1 cysteyl (3-H), 3.04-3.16 (2 H, m, 2 lysyl EH), 3.09 (1 H, dd, Ja, p 4.9 and Jp, p ~ 14, 1 cysteyl (3-H), 3.58 and 3.59 (2 H, 2 dd, J3.4 4.6 and J4.5 9.2, 2 4-H), 3.73-3.76 (2 H, m, 2 glycyl a-H), 3.83-3.88 (2 H, m, 2 5-H), 4.17 (1 H, m, lysyl aH), 4.25-4.29 (2 H, m, 2 3-H), 4.52 (1 H, m, 1 cysteyl aH), 6.22 and 6.28 (2 H, 2 br d, J2.3 4.0, 2 2-H), 7.86 (1 H, t, JE, rrH 5.4, 1 lysyl ENH), 8.02 (1 H, t, Ja, NH 5.6, 1 glycyl NH), 8.06 (1 H, d, Ja, NH 6.6, 1 lysyl ° 'NH), 8.32 (1H, d, Ja, NH 7.7, 1 cysteyl NH);
NMR spectrum ~ 3C: 8o (8I.3 MHz; H20-DZO 90:10) 22.9 (lysyl yC), 28.3 (lysyl b-VS), 29.4 (C (CH3) 3), 30.7 (lysyl (3-C), 31.7 and 31.5 (2 6-C), 39.8 (lysyl sC), 40.6 (cysteyl (3-C), 41.8 (glycyl aC), 48.7 (C (CH3) 3), 53.3 (cysteyl aC), 54.9 (lysyl a-C), 66.3 and 66.8 (2 4-C and 2 5-C), 71.9 and 72.0 (2 3-C), 131.1 and 132.1 (2 1-C), 133.1 and 133.7 (2 2-C), 170.5, 171.0, 172.7, 173.4 and 174.6 (S XC = O);
TOF-PDMS spectrum: m / z 729 (M + Na) +.
The coupling constants between the 3-H / 4-H and 4-H / 5-H protons of two (-) - shikimic acid residues present in compound 1 such that determined by 1 H NMR spectrometry are 4.6 and 9.2 Hz respectively. These constants are compatible with a pseudo-equatorial orientation of the hydroxyl functions litters by the carbon atoms located in positions 4 and 5 of said acids and are coming confirm the structural analogy between (-) - shikimic acid and D-mannose.
b) Preparation of compound 3 Compound 3 was prepared on a 2 mmol scale using (using a Fmoc-glycine o-methoxy p- (benzyloxy) benzyl resin (available under the trade name Fmoc-Gly-Sasrin with the company BACHEM) responsible at 0.8 mmol / g and using ~ an Fmoc / tert-butyl strategy for the protection of amino acids couple, ~ a mixture of HBTU / DIEA in NMP, as activating reagents for the amino acid couplings, ~ a piperidine / NMP mixture to remove the Fmoc protective groups, and ~ Ninhydrin and TNBS tests to control the operations of coupling and deprotection.
The couplings of S- (tert-butylthio) -L-cysteine and L-lysine were carried out under conditions identical to those used for the preparation of compound 1.
On the other hand, the coupling of the two molecules of (-) - quinic a was carried out according to a different operating protocol from that used for the coupling of (-) - shikimic acid. Indeed, it turned out that the activation acid (-) - quinique immediately resulted in the formation of a y-lactone bicyclic, by esterification between the carboxyl group carried by the carbon located in position 1 and the hydroxyl group carried by the carbon located in position 3 of this acid, and it was therefore necessary to protect the hydroxyl functions of ((-) - quinic acid before proceed to its coupling.
It was therefore chosen to subject (-) - quinic acid to a peracetylation with acetic anhydride, so as to obtain (acid (ls ", 3R, 4s", SR) -1,3,4,5-tetraacetoxycyclohexane-1-carboxylic which is none other that the tetraacetylated derivative of (-) - quinic acid and which will be referred to as compound 9 in what follows, then transform the latter into its acid fluoride (called compound

18 in the following) by treating compound 9 with cyanuryl fluoride.
To do this, a drop of perchloric acid was added, to at room temperature and with stirring, to a suspension containing 4 g (21 mmoles) (-) - quinic acid in 30 ml of a mixture of acetic acid / Ac20 (2: 1).
The agitation has continued for 12 hours after which the resulting mixture has been diluted with chloroform and extracted with a saturated solution in sodium bicarbonate, then with water. The organic layer was dried by of sodium sulfate, filtered and concentrated under reduced pressure. The residue has been rushed by addition of pentane to give 6.45 g of compound 9, ie a yield of 86%.
The transformation of acid 9 into its fluoride was carried out in adding, drop by drop and with stirring, 937 p, 1 (11.10 mmol) of fluoride cyanuryl to a solution comprising 500 mg (1.39 moles) of compound 9 in 6 ml of methylene chloride and 112 u1 (1.39 mmol) of pyridine, and carrying the mixed resulting at reflux for 2 hours. A white precipitate having formed, this mixture was filtered, then it was extracted with water. Elimination of solvent for the organic layer, after drying of the latter with sodium sulfate, has leads obtaining an oil corresponding to compound 18 and which has been subjected to a vacuum drying for several hours.

Compounds 9 and 18 have spectral characteristics following Compound 9 ~ ect NMR 'H: 8H (300 MHz; CDCl3) 1.87 (1 H, dd, JS, ~ 10.2 and J6,6 ~ 13.6, 6-H), 1.96, 1.98, 2.03 and 2.08 (4 x 3 H, 4 s, 4 CH3), 1.98-2.03 (1 H, m, 6'-H), 2.36 (1 H, dd, JZ, 2 ~ 15.9 and JZ, 3 3.3, 2-H), 2.50 (1 H, dd, J2,2 ~ 15.9 and JZ ~, ~ 3.2, 2'-H), 4.97 (1 H, dd, J3, a 3.4 and J4.5 9.4, 4-H), 5.31 (1 H, ddd, J4.5 9.4, J5.6 10.2 and JS, ~~ 4.2, 5-H), 5.50 (1 H, m, 3-H);
NMR spectrum ~ 3C: S ~ (81.3 MHz; CDC13) 21.0, 21.1, 21.2 and 21.4 (4 CH3), 32.0 and 36.3 (2-C and 6-C), 66.7, 67.7 and 71.3 (3-C, 4-C and SC), 78.8 (1-C), 170.1-170.5 (4 COZMe), 173.1 (COZH);
Compound 18 1 H NMR spectrum: 8H (300 MHz; CDCl3) 1.97-2.05 (1 H, m, 6-H), 2.03, 2.06, 2.08 and 2.16 (4 x 3 H, 4 s, 4 CH3), 2.45 (1 H, dd, J2.2 ~ 15.4 and J2.3 3.7, 2-H), 2.50-2.66 (2 H, m, 2'-H and 6'-H), 5.08 (1 H, dd, J3.4 3.5 and J4s 9.1, 4-H), 5.38 (1 H, ddd, Ja, s 9.1, J5.6 ~ 9.1 andJS, ~ 5.1, 5-H), 5.56 (1 H, m, 3-H);
13C NMR spectrum: 8 (81.3 MHz; CDCl3) 20.4, 20.6, 20.8 and 20.9 (4 CH3), 31.9 and 35.4 (2-C and 6-C), 66.0, 68.2 and 70.4 (3-C, 4-C and 5-C), 76.7 (J ~, F 53.2, 1-C), 160.3 (J ~ F
373.4, COF), 169.6, 169.7, 169.8 and 169.8 (4 COZMe).
The coupling of the two molecules of compound 18 was carried out in acylating the resin a first time by 2 equivalents (per amine function in front of react) of compound 18 in the presence of 3 equivalents of DIEA in NMP
(duration reaction time: 60 minutes), then, after washing the resin with NMP
(3 x 2 minutes) and DCM (3 x 2 minutes), by facylating a second time, but by using 1 equivalent of the different reagents.
At the end of this coupling operation, the solvent was filtered, the resin was washed successively with NMP (3 x 2 minutes) and DCM (3 x 2 minutes), then dried.
The cleavage of compound 3 was obtained by treating the resin with 20 ml of a CHZCIz / TFAfPr3SiH mixture (95: 2: 3) (4 x 5 minutes). The solution was concentrated under reduced pressure and the residue thus obtained was dissolved in of skin, then subjected to lyophilization leading to the production of 1.40 g of a gross compound which was dissolved in 25 ml of methanol. To the solution thus obtained, was added, dropwise and with stirring, 1 M methanolic sodium methylate until obtaining a pH of about 9, then this mixture was allowed to react for 3 hours, by monitoring the course of the reaction by RP-HPLC. The mixture was then concentrated under reduced pressure. A 100 mg sample of the residue has summer purified by semi-preparative RP-HPLC [gradient (AB): 100: 0 to 90:10 in 10 minutes, then 90:10 to 50:50 in 70 minutes] to lead to obtaining, after lyophilization, 41 mg of pure compound 3, ie a yield of 59%.
Compound 3 has the following characteristics Ontic rotation: [a] D -73 (c 0.49, Hz0);
Spectrum I ~ MN IH: 8H (300 MHz; H20-D20 90:10) 1.12 (3 x 3 H, s, 3 CH3), 1.15-1.22 (2 H, m, 2 lysyl yH), 1.34 (2 H, quintet, JY, s 7.2 and Js, E 7.2, 2 lysyl 8-H), 1.57-1.90 (10 H, m, 2 lysyl ~ iH, 4 2-H and 4 6-H), 2.83 (1 H, dd, Ja, p 8.6 and Jp, ~~ 14, 1 cysteyl (3-H), 3.02 (3 H, m, 1 cysteyl [3-H and 2 lysyl sH), 3.33 (2 H, dd, J3.4 9.6 and Ja, s 2.9, 2 4-H), 3.79 (2 H, d, Ja, NH 5.8, 2 glycyl aH), 3.80-3.90 (2 H, m, 2 5-H), 4.01 (2 H, br s, 2 3-H), 4.11 (1 H, dt, Ja, NH 7.0 and Ja, p 7.2, 1 lysyl aH), 4.54 (1 H, dt, Ja, NH
7.6 and JQ, ~ 6.0, 1 cysteyl aH), 8.02 (1 H, t, J ° "NH 5.9, 1 lysyl ENH), 8.11 (1 H, d, Ja, rrH 7.0, 1 lysyl ° 'NH), 8.21 (1 H, t, Ja, NH 5.8, 1 glycyl NH), 8.35 (1 H, d, Ja, NH
7.6, 1 IVH cysteyl);
~ 3N NMR ect: 8 (81.3 MHz; Hz0-D20 90:10) 22.7 (lysyl yC), 28.3 (lysyl SC), 29.4 (C (CH3) 3), 30.9 (lysyl (3-C), 37.5 (2 2-C), 39.4 (lysyl sC), 40.7 (cysteyl [3-C and 2 6-C), 41.8 (glycyl aC), 48.6 (C (CH3) 3), 53.1 (cysteyl aC), 54.4 (lysyl aC), 66.7 and 66.8 (2 3-C), 70.8 (2 5-C), 75.4 (2 4-C), 77.2 (2 1-C), 172.5, 173.4, 174.4, 177.1 and 177.4 (S XC = O);
ESI-MS spectrum: m / z 741 (MH) -.
The coupling constants between the 3-H / 4-H and 4-H / 5-H protons of two acid (-) - quinic residues present in compound 3 such that determined by 1 H NMR spectrometry are 9.6 and 2.9 Hz respectively. These constants are compatible with a “chair” conformation of cyclohexane placing the group amide in equatorial position, hydroxyls 3 and 4 in trans di-equatorial and hydroxyls 4 and 5 in equatorial-axial cis relationship.

1.3 - Couplings of the A 'dendrimers and of the tripentides carrying the remains (-) - shikimic and (-) - quinic acid The costs of compounds 13, 14 and 15 and of compounds 1 and 3 have were carried out according to a procedure comprising 2 stages, namely ~ a first step aimed at reducing the disulfide bond (S-SBu ') that present each of compounds 1 and 3, treating these compounds with tri-n-butylphosphine in a degassed n-propanol / water mixture (50:50), so as to obtain elimination of the tert-butylthiol protective group carried by each of the compounds 1 and 3 and therefore deprotection of their thiol function, and ~ a second step aimed at reacting the compounds 1 and 3 thus reduced with the compounds 13, 14 and 15, in a DMF / degassed water mixture (90:10) and in the presence of potassium carbonate, so as to obtain the formation of a bond thioether between the thiol function of compounds 1 and 3 and one of the functions chloroacetyls compounds 13, 14 and 15.
So, to a solution containing 1.5 equivalent (per group chloroacetyl to react in the second step) of compound 1 or compound 3 in 1 ml of a degassed n-propanol / water mixture (50:50), 1 equivalent of tri-n-butylphosphine. Each reaction mixture was left with stirring and under nitrogen, at room temperature for 24 hours, then the solvents and tert-butylmercaptan released were evaporated under reduced pressure and the residue was dried under vacuum for 15 minutes.
Each of these residues, after dissolution in a DMF / water mixture degassed (90:10), was added to 1 equivalent (1-5 p, moles) of a compound 13, 14 or 15 and 20 equivalents of potassium carbonate. Each reaction mixture was leash, with stirring and at room temperature, for 72 to 96 hours - the reactions being controlled by RP-HPLC - then diluted in water and lyophilized. The residues have were then purified by semi-preparative RP-HPLC.
Were so obtained ~ 3.39 mg of compound 21 (yield: 61%) using a gradient (A / C): 100: 0 to 70:30 in 70 minutes, ~ 5.98 mg of compound 22 (yield: 66%) using a gradient (A / C): 100: 0 to 75:25 in 25 minutes, then isocratic, ~ 2.25 mg of compound 23 (yield: 42%) using a gradient (A / C): 100: 0 to 80:20 in 35 minutes, then isocratic, ~ 5.75 mg of compound 24 (yield: 56%) using a gradient (A / C): 100: 0 to 75:25 in 25 minutes, then isocratic, ~ 3.15 mg of compound 25 (yield: 43%) using a gradient (A / C): 100: 0 to 75:25 in 40 minutes, then isocratic, and ~ 7.03 mg of compound 26 (Yield: 61%) using a gradient (A / C) 100: 0 to 80:20 in 35 minutes, then isocratic, all in the form of a white powder.
Compounds 21 to 26 have spectral characteristics following Compound 21 NMR spectrum ~ H
8H (600 MHz; Hz0-DZO 90:10) 1.31-1.46 (8 H, m, 8 lysyl yH), 1.43-1.60 (6 H, m, lysyl 8-H), 1.64 (2 H, m, 2 lysyl 8-H), 1.68-1.89 (8 H, m, 8 lysyl ~ 3-H), 2.17-2.23 (4 H, m, 4 6-H), 2.46 and 2.51 (2 x 1 H, 2 dt, Ja, a ~ 15.4 and Ja, p 6.2, 2 alanyl a-H), 2.70-2.80 (4 H, m, 4 6'-H), 2.89-2.95 (2 H, m, 2 cystéyl ~ 3-H), 2.96-3.01 (2 H, m, 2 lysyl s-H), 3.06 (2 H, dd, Ja, p 5.3 and, Ip, p ~ 14.9, 2 cysteyl Vii'-H), 3.21 (2 H, m, 2 lysyl sH),), 3.25 (2 H, m, 2 lysyl sH), 3.27 (1 H, d, J 16.5, CHH), 3.30 (1 H, d, J 16.5, CHI, 3.33 (2 H, s, CHZ), 3.41-3.50 (2 x 1 H, 2 ddt, Ja, p 6.2, JR, p ~ 12.6 and Jp, NH
6.2, 2 (3-alanyl [3-H), 3.71-3.76 (4 H, m, 4 4-H), 3.95 (4 H, d, Ja, NH 6.3, 4 glycyl a-H), 4.00-4.05 (4 H, m, 4 5-H), 4.25 (2 H, dt, Ja, p 5.6 and Ja, rrH 6.3, 2 lysyl aH), 4.32-4.38 (4 H, m, 4 lysyl aH), 4.42-4.44 (4 H, m, 4 3-H), 4.58-4.62 (2 H, m, 2 cysteyl a-H), 6.38 and 6.44 (2 x 2 H, 2 br d, JZ, 3 4.0, 2 x 2 2-H), 6.90 (1 H, s, NHFi), 7.54 (br s, ENH2), 7.60 (1 H, s, NHI ~, 8.07 (2 H, t, Ja, NH 6.8, 2 lysyl ENH), 8.15 (1 H, t, Ja, NH 6.2, (3-alanyl NH), 8.24 (1 H, t, J ~, HH 6.3, 1 lysyl ENH), 8.25-8.27 (2 H, m, 2 lysyl ° 'NH), 8.31 and 8.32 (2 x 1 H, 2 t, Ja, NH 6.3, 2 glycyl NH), 8.42 (1 H, d, Ja, NH 6.8, 1 lysyl "NH), 8.47 (1 H, d, Ja, NH 6.3, 1 lysyl ° 'NH), 8.61 (2 H, d, Ja, NH 7.6, 2 cysteyl NH);
ESI-MS spectrum: m / z 1660.4 (MH) -, 829.9 (M-2H) Z '.

Compound 22 1 H NMR spectrum 8H (300 MHz; HZO-D20 90:10) 1.22-1.28 (8 H, m, 8 lysyl yH), 1.34-1.43 (6 H, m, lysyl 8-H), 1.52 (2 H, m, 2 lysyl 8-H), 1.57-1.64 (8 H, m, 8 lysyl (3-H), 1.65-1.99 (16 H, m, 8 2-H and 8 6-H), 2.33 (2 H, t, Ja, p 6.3, 2 ~ i-alanyl aH), 2.69-2.84 (4 H, m, 2 cysteyl (3-H and 2 lysyl EH), 2.92 (2 H, dd, Ja, p 5.3 and Jp, p ~ 14, 1 2 cysteyl (3-H), 2.98-3.11 (6 H, m, 6 lysyl sH), 3.15 and 3.19 (2 x 2 H, 2 s, 2 CHZ), 3.23 and 3.34 (2 x 1 H, 2 ddt, Ja, p 5.3, Jp, p ~ 13.0 and Jp, NH 5.7, 2 (3-alanyl (3-H), 3.38 (4 H, dd, J3.4 3.1 and Ja, S 9.7, 4 4-H), 3.76 and 3.77 (4 H, 2 t, JQ, NH 6.3, 2 glycyl aH), 3.87-3.96 (4 H, m, 4 5-H), 4.06 (4 H, br s, 4 3-H), 4.10-4.16 (4 H, m, 4 lysyl aH), 4.60-4.64 (2 H, m, 2 cysteyl ~ iH), 6.73 and 7.42 (2 x 1 H, 2 s, NHH and NHl ~, 7.96 (1 H, t, Ja, ~
5.7, (3-alanyl NH), 8.07 (1 H, t, Ja, NH 6.3, 1 lysyl ENH), 8.10 (1 H, t, Ja, NH 6.9, 1 lysyl ENH), 8.11 (2 H, t, Ja, NH 6.3, 2 glycyl NH), 8.12 (1 H, t, JE, NH 6.8, 1 lysyl ~ NH), 8.13 (2H, d, Ja, NH 6.8, 2 lysyl "NH), 8.25 (1 H, d, Ja, NH 6.7, 1 lysyl ° 'NH), 8.29 (1 H, d, Ja, rrH 6.3, 1 lysyl ° 'NH), 8.39 (2H, d, Ja, NH 7.6, 2 cysteyl NH;
ESI-MS spectrum: m / z 1732.5 (MH) ', 865.6 (M-2H) 2'.
Compound 23 ESI-MS spectrum: measured: 3235.0, calculated: 3235.6, m / z 1616.4 (M-2H) 2-, 1077.3 (M-3H) 3-, 807.7 (M-4H) 4 '.
Compound 24 ESI-MS spectrum: measured: 3379.0, calculated: 3379.7, m / z 1688.3 (M-2H) z ', 1125.3 (M-3H) 3 ', 843.8 (M-4H) 4'.
Compound 25 ESI-MS spectrum: measured: 6382.0, calculated: 6383.0, m / z 1594.5 (M-4H) 4 ', 1275.5 (M-SH) 5 ', 1062.8 (M-6H) 6-, 910.8 (M-7H)''.
Compound 26 ESI-MS spectrum: measured: 6671.0, calculated: 6671.2, m / z 2221.9 (M-3H) 3 ', 1666.7 (M-4H) 4 ', 1333.1 (M-SH) 5', 1110.8 (M-6H) G ', 952.0 (M-7H)''.

EXAMPLE 2. PREPARATION OF LIGANDS CONFORMING TO
THE INVENTION
Four ligands - which will be called ligands 21F, 22F, 23F and 24F
in the following - and corresponding to the general formula (III) in which ~ m represents an integer equal to 4 or 8, B 1 represents an acid (-) - shikimic or (-) - quinic residue, ~ n is an integer equal to 2 or 4 (in the case where m is respectively equal to 4 and 8), . X represents the remainder of a tripeptide of sequence (S 1) as defined in example 1, A represents the remainder of a dendrimer A 'comprising 2 or 4 (in the case where m East equal to 4 and 8 respectively) L-lysine residues and having, on the one hand, 2 or chloroacetyl groups for its binding with an equivalent number of tripeptides and, on the other hand, a free amine group capable of allowing its bond with Y, . Y represents the remainder of a fluorescein molecule, M represents an amide group (-CO-NH-), N represents a thioether group (-CHZ-S-CHZ-CO-), while P represents a thiourea group (-NH-CS-NH-), were prepared by coupling compounds 1 and 3, after deprotection of their function thiol under conditions identical to those described in ~ 1.4 of said example 1, with compounds 13 and 14 which have previously reacted with FITC.
Thus, as described in ~ 1.4 of Example 1, to a solution containing 1.5 equivalents (per chloroacetyl group to react later) from compound 1 or compound 3 in 1 ml of a degassed n-propanol / water mixture (50:50), 1 equivalent of tri-n-butylphosphine was added. Each reaction mixture has been left, with stirring and under nitrogen, at room temperature for 24 hours and then the solvents were evaporated under reduced pressure and the residue was dried under vacuum for 15 minutes.
In addition, 1 equivalent (1-2.5 p, moles) of compound 13 or 14 was reacted with 1.1 equivalents of FITC and 4 equivalents of isopropylethylamine in 1 ml of degassed DMF, at room temperature and (darkness, for 4 hours at the end of which each reaction mixture was added to a solution comprising one of the residues previously obtained in 300 μl of degassed water.
The walls of vessels containing said reaction mixtures have been rinsed with 200 ~ .l of degassed DMF and the pH of the various reaction mixtures was brought to a value of 8-8.5 by addition of potassium carbonate.
These reaction mixtures were then left in the dark.
for 48 hours, the reactions being controlled by RP-HPLC. Then, each reaction mixture was diluted in water containing 10% acetic acid and lyophilized and the residues were purified by semi-preparative RP-HPLC.
Were so obtained ~ 1.82 mg of ligand 21F (Yield: 46%) using a gradient (A / C): 100: 0 to 75:25 in 15 minutes, then 75:25 to 65:35 in 30 minutes, ~ 1.42 mg of ligand 22F (Yield: 27%) using a gradient (A / C): 100: 0 to 75:25 in 15 minutes, then 75:25 to 65:35 in 30 minutes, ~ 4.56 mg of ligand 23F (Yield: 62%) using a gradient (A / C): 100: 0 to 85:15 in 15 minutes, then 85:15 to 70:30 in 30 minutes, ~ 3.57 mg of ligand 24F (Yield: 50%) using a gradient (A / C): 100: 0 to 85:15 in 15 minutes, then 85:15 to 70:30 in 30 minutes, all in the form of a white powder.
The chemical structures of these ligands are shown on the Figures 3 and 4.
Their ESI-MS spectra are as follows ~ ligand 21F: m / z 2123.5 (MH) -, 1062.3 (M-2H) Z ' ~ ligand 22F: measured: 2051.0, calculated: 2051.2, m / z 1026.1 (M-2H) Z ' ~ 23F ligand: measured: 3769.0, calculated: 3769.1, m / z 1254.9 (M-3H) 3 ', 941.2 (M-4H) ° -~ 24F ligand: measured: 3624.0, calculated: 3624.1, m / z 1207.0 (M-3H) 3-, 905.4 (M-4H) 4-.
EXAMPLE 3. PREPARATION OF LIGANDS CONFORMING TO
THE INVENTION
Two ligands corresponding to the general formula (III) in which ~ m is an integer equal to 3, ~ B ~ represents an acid (-) - shikimic or (-) - quinic residue, ~ n is an integer equal to 0 (so that X is absent), . A represents the remainder of a linear sequence A 'comprising 3 residues L-lysine and having 3 free amine groups for its coupling with the three molecules (-) - shikimic or (-) - quinic acid and being, on the other hand, linked to Y via a bond amide, Y represents a compound of interest formed by three Arg-Gly-Arg residues linked to a class II epitope, in this case chicken ovalbumin peptide 323-339 (who will be referred to as Ova 323-339 below), M and P represent an amide group (-CO-NH-), have been prepared.
The preparation of these ligands was carried out by proceeding ~ first, the synthesis, on a solid support, of a peptide presenting the next sequence (S2) Ac-Cys [S- (tert-butylthio)] - Lys (Mtt) -Lys (Mtt) -Lys (Mtt) -Arg (Pmc) -Gly-Arg (Pmc) -Ile-S er ('Bu) -Gln (Trt) -Ala-V al-His (Trt) -Al a-Ala-His (Trt) -Ala-Glu ('Bu) -Ile-Asn (Trt) -Glu (' Bu) -Ala-Gly-Arg (Pmc) (S2) in which - the 17 amino acid residues located at the C-terminal end correspond to the sequence of the Ova 323-339 peptide, - the group formed by the 3 Arg-Gly-Arg residues located upstream of the sequence of said Ova 323-339 peptide represents a group sensitive to endopeptidases, - the group formed by the 3 Lys-Lys-Lys residues corresponds to (chain of L-lysine, while - the residue S- (tert-butylthio) -L-acetylated cysteine located at the N end terminal is intended to allow subsequent coupling of the ligand, for example for obtaining a dimer;
~ then, when coupling the L-lysine residues of this peptide and acids (-) -shikimic and (-) - quinic (in the form of its tetra-O-acetylated derivative), by formation of a amide bond between the s-amine groups of said L-lysine residues and the groups carboxyls of said acids.
These ligands will be called ligand 25 and ligand 28 respectively.
in the following.
The peptide of sequence (S2) has been synthesized on a scale of 0.25 mmol from 337 mg of an MBHA Rink amide resin loaded with 0.74 mmol / g (SENN CHEMICALS Company), and using - an Fmoc / tert-butyl strategy for the protection of amino acids couple, a 2,2,2,5,7,8-pentamethylchroman-6-sulfonyl group as a group protector of the side chains of L-arginines, - a tert-butyl group as a protective group for the side chains of L-serine and L-glutamic acids, - a trityl group as a protective group for the side chains of the L-asparagine, L-glutamine and L-histidines, - a 4-methyltrityl group as a protective group for the side chains of L-lysines, - a mixture of HBTU / HOBt / DIEA in DMF, as activation reagents for the coupling of amino acids, a piperidine / DMF mixture to eliminate the Fmoc protective groups, and - Ninhydrin and TNBS tests to control the operations of coupling and deprotection.
The couplings of the first 20 amino acids were carried out from automatically using an APPLIED BIOSYSTEMS ABI431 synthesizer, while the couplings of L-lysines and S- (tert-butylthio) -L-cysteine have been performed manually.
In all cases, these couplings were carried out using 4 equivalents (per amine function to react) of amino acid, in the presence of HBTU / HOBtIDIEA mixture (4: 4: 12 eq per amine function to react) in DMF, and activating the amino acids for 1 minute before adding them to the resin. In practice, the HBTU (4 eq.) Was dissolved in DMF, then added to a solution containing (amino acid to be coupled, HOBt (4 eq.) and DIEA (8 eq.) in some DMF. The whole was mixed for 1 minute, then added to the resin previously soaked with DMF. The resulting reaction mixture was stirred mechanically throughout the duration of the reaction, i.e. for 40 minutes for automated pairings and 45 minutes for manual pairings.
The couplings of L-lysines and S- (tert-butylthio) -L-cysteine have all were followed by a treatment of the resin with an Ac20 / DIEA / DCM mixture (5:10:85) with a duration of 10 minutes intended to acetylate the amino groups which would not have been paired.
In addition, between each coupling operation, the resin was successively filtered, washed with DMF (3 x 2 minutes) and DCM
(3 x 2 minutes), treated with a piperidine / DMF mixture (20:80) to obtain the cleavage Fmoc groups, and washed again with DMF (2 x 2 minutes) and DCM
(3 x 1 minute).
A (from the coupling of S- (tert-butylthio) -L-cysteine and its acetylation, the 4-methyltrityl protecting groups of L-lysine residues have summer hydrolyzed by washing the resin with a TFA / DCM mixture (1:99) in continuous flow until its yellow color disappears.
Couplings of the peptide of sequence (S2) with acids (-) - shikimique and (-) - quinique were made by reacting, to ambient temperature and for 45 minutes, fractions of the resin thus washed with 4 acid equivalents (-) - shikimic or 4 equivalents of tetra-O-acetyl acid - (-) - quinic beforehand activated by 4 equivalents of HBTU and 4 equivalents of HOBt, in the presence of 12 equivalents of DIEA in DMF, and by monitoring reactions by tests to the nynhidrine and TNBS.
The cleavage of the resulting products and their deprotection have been carried out by treating the resin with 10 ml of a TFA / H20fPr3SiH mixture (95: 2.5: 2.5) with gram of resin, at room temperature and for 90 minutes. After what, they were successively precipitated in cold diethyl ether, centrifuged, dissolved in Skin, lyophilized and purified by semi-preparative RP-HPLC
[gradient (A / B): 100: 0 to 50:50 in 120 minutes].
Were so obtained ~ 48 mg of ZS ligand (Yield: 29%) using a gradient (A / B): 100: 0 to 50:50 in 100 minutes; and ~ 25 mg of compound 26 (Yield: 15%) using a gradient (A / B): 100: 0 to 60:40 in 100 minutes.
This compound 26 was subjected to deacylation to obtain the ligand 28. To do this, 23 mg of compound 28 dissolved in 10 ml of methanol distilled over magnesium, 300 μl of a 1M solution of methanolate sodium in methanol. The reaction mixture was stirred at temperature ambient, under nitrogen, for 45 minutes and the reaction was stopped by the addition of a few drops of acetic acid. The reaction crude was concentrated under pressure reduced and has been purified by RP-HPLC using a gradient (AB): 100: 0 to 80: 20 in 30 minutes, then 80: 20 to 70: 30 in 25 minutes, then isocratic for 10 minutes for lead to 15 mg of ligand 28 (yield: 75%).
The ESI-MS spectra of ligands 25 and 28 are as follows ~ ligand, 25: measured: 3228.0, calculated: 3228.7, m / z 1076.9 (M + 3H) 3+, 808.0 (M + 4H) '+, 646.7 (M + SH) 5+
~ ligand 28: measured: 3283, calculated: 3282.7, m / z 1641.9 (M + 2H) z +, 1094.9 (M + 3H) 3+, 821.4 (M + 4H) 4+, 657.4 (M + SH) 5+.
EXAMPLE 4. PREPARATION OF LIGANDS CONFORMING TO
THE INVENTION
Two ligands corresponding to the general formula (III) in accordance with the invention (Figure 5), in which ~ m is an integer equal to 2, ~ n is an integer equal to 1, ~ y is an integer equal to 1, ~ B ~ represents an acid (-) - quinic (ligand 33F) or (-) - shikimic residue (ligand 34F), ~ A represents the remainder of a dipeptide comprising 1 L-lysine residue and 1 residue (3-alanine (No. '- (chloroacetyl) -L-lysyl- ~ 3-alanine-amide, of which the chemical structure is shown in Figure 5), ~ X represents a tripeptide of formula (S 1), as defined in example 1, . Y represents the remainder of a fluorescein molecule, ~ M, N and P respectively represent an amide group (-CO-NH-), thioether (-CHZ-S-CHz-CO-) and thiourea (-NH-CS-NH-), have been prepared.
a) Preparation of the dipeptide Na- (chloroacetyll-L-lysyl-Q-alanine-amide.
This dipeptide is synthesized manually using a Rink resin amide-AM-PS (0.70 mmol / g, Senn Chemicals) using an Fmocltert-butyl, on a scale of 0.7 mmole. The coupling is followed by a TNBS test : of HBTU dissolved in NMP (4 eq.), Is added to a mixture of the amino acid (4 eq.), HOBt (4 eq.) and DIEA (8 eq.) in the NMP. After 1 minute of agitation, the reaction mixture is added to the peptidyl-resin (1 eq.) beforehand soaked with NMP comprising DIEA (4 eq.), And stirred for 40 minutes. Peptidyl-resin (i.e. the peptide sequence on solid support) is filtered, washed with NMP (3 x 2 minutes) and CHZC12 (3 x 2 minutes). Protective groups Fmoc are suppressed by treatment with 20% piperidine in NMP
(1 x 2 minutes, 1 x 20 minutes), followed by washing with NMP (2 x 2 minutes) and of CHZCIz (2 x 1 minutes). After the second coupling step, the peptidyl-resin East deprotected and acylated using an excess (4 eq.) of chloroacetic anhydride, prepare via DIC activation. Finally, the dipeptide is separated from the support and deprotected under the action of a TFA-TIS-HZO 90: 5: 5 mixture (11 ml per gram of peptidyl-resin), for 1.5 h at room temperature, precipitated in a diethyl ether mixture-heptane (1: 1) cold, centrifuged, dissolved in water and lyophilized. Dipeptide (184 mg, 65%) is obtained in the form of a white powder after purification by RP-HPLC
(with the following gradient: 100: 0 to 90:10 (A / B) in 20 minutes), followed by a lyophilization.
Its analysis is as follows: TOF-PDMS m / z 292.6 (M + H) +; 1 H NMR (D20-H20 10:90): 8 8.37 (d, J = 6.8 Hz, 1 H, lysyl "NH), 8.07 (t, J = 5.7 Hz, 1 H, (3-alanyl NH), 7.40 (broad s, 1H, NHZ), 4.11 (dt, J = 6.8 and 6.8 Hz, 1H, lysyl aH), 4.01 (s, 2H, CHZ), 3.31 (dt, J = 5.7 and 6.6 Hz, 2H, (3-alanyl (3-H), 2.84-2.81 (m, 2H, lysyl E-H), 2.33 (t, J
= 6.6 Hz, 2H, ~ 3-alanyl aH), 1.72-1.53 (m, 4H, lysyl (3- and 8-H), 1.42-1.22 (m, 2H, lysyl yH); NMR 13C (D20-Hz0 10:90): S 175.7, 172.4, and 168.6 (3 COIS, 53.2 (lysyl aC), 41.0 (CHZ), 38.4 (lysyl sC), 34.8 and 33.5 ((3-alanyl ci-C and ~ iC), 29.4 (lysyl (3 C), 25.3 (lysyl 8-C), 21.0 (lysyl yC).

b) Obtaining the ligand ds corresponding to the general formula (III
according to the invention.
The coupling of the dipeptide prepared in a) with, on the one hand, the tripeptides carrying residues (-) - shikimic (compound 1) and (-) - quinic (compound 3), the synthesis of which is described in Example 1, and on the other hand, with a molecule fluorescein, is carried out in accordance with the protocol described in Example 2.
We thus obtains ligands 33F and 34F.
5.04 mg (80% yield) of ligand 33F were obtained under the form of a white powder, after purification by RP-HPLC (gradient: 100: 0 to 85:15 (A / C) in 15 minutes, then 85:15 to 75:25 (A / C) in 30 minutes, then gradient isocratic), followed by lyophilization. Analysis of ligand 33F: ESI-MS
positive m / z 1300.3 (M + H) +, 650.8 (M + 2H) 2+; 1 H NMR (DMSO-db): 8 10.2 (broad s, 1H, NH-fluorine), 8.40-8.32 (m, 3H, H-fluorine, lysyl ° 'NH et lysyl ~ NH), 8.31 (t, J = S.1 Hz, 1 H, glycyl NH), 8.24 (d, J = 7.8 Hz, lysyl "NH), 8.10 (t, J = 5.2 Hz, 1 H, (3-alanyl NH), 7.82-7.80 (m, 2H, cysteinyl NH and lysyl ENH), 7.80 (d, J = 8.3 Hz, H-fluorine), 7.41 (broad s, 1H, NHH), 7.22 (d, J = 8.3 Hz, H-fluorine), 6.90 (broad s, 1H, NHF ~, 6.74-6.60 (m, 6H, 6H-fluorine).
2.42 mg (80% yield) of ligand 34F were obtained under the form of a white powder, after purification by RP-HPLC (gradient: 100: 0 to 85:15 (A / C) in 15 minutes, then 85:15 to 70:30 (A / C) in 50 minutes), followed of a lyophilization. Analysis of the 34F ligand: positive ESI-MS m / z 1264.4 (M + H) +, 632.8 (M + 2H) 2+; ~ 'H (DMSO-d6): 8 10.07 (s, 1H, OH), 9.81 (s broad, 1H, NH-fluorine), 8.22 (t, J = 5.7 Hz, 1 H, glycyl NH), 8.20 (broad s, 1 H, H-fluorine), 8.11 (d, J
= 8.0 Hz, 2H, 2 lysyl aNH), 8.01 (t, J = 5.6 Hz, 1H, lysyl ENH), 7.99 (t, J = 5.8 Hz, (3-alanyl NH), 7.77 (t, J = 5.6 Hz, 1 H, lysyl ENH), 7.75 (d, J = 7.6 Hz, 1 H, cysteinyl NH), 7.69 (d wide, J = 8.3 Hz, H-fluorine), 7.28 (s broad, 1H, NHH), 7.12 (d, J = 8.3 Hz, H-fluorine), 6.80 (s wide, 1H, NHI ~, 6.74-6.60 (m, 6H, 6 H-fluorine), 6.31 and 6.22 (2 s large, 2 shikimoyl H-2).

EXAMPLE 5. PREPARATION OF LIGANDS CONFORMING TO
THE INVENTION
Four ligands corresponding to the general formula (III) conforming to the invention, in which ~ m is an integer equal to 8, ~ n is an integer equal to 4, ~ y is an integer equal to 1, ~ B ~ represents an acid (-) - shikimic or (-) - quinic residue, ~ Y represents the remainder of a fluorescein molecule, . A represents the remainder of a dendrimer A 'comprising 4 L-lysine residues and having 4 chloroacetyl groups for its bond with 4 tripeptides, and a group amine, free suitable for allowing its bond with Y, ~ X represents the remainder of a tripeptide of sequence (S2) or (S3), such that they will be defined below, . M, N and P respectively represent an amide group (-CO-NH-), thioether (-CHz-S-CHZ-CO-) and thiourea (-NH-CS-NH-), were prepared according to the following protocol.
a) Preparation of compound Q, derived from 4uinic acid in which the hydroxyls located in positions 3 and 4 are protected in the form of a diacetal Fi ure 6.
Compound Q, namely the acid (1s °, 3R, 4s ", 5R, 2'S, 3'S) -3,4-D-(2 ', 3' -dimethoxybutane-2 ', 3'-diyl) -1,3,4,5-tetrahydroxycyclohexane-1-carboxylic, is obtained from 980 mg (3.06 mmol) of the methyl ester of the acid 3,4-0-(2 ', 3'-dimethoxybutane-2 ', 3'-diyl) -1,3,4,5-tetrahydroxycyclohexane-1-carboxylic (described by JL MONTCHAMP et al. in J. Org. Chem., 1996, 61, 3897-3899) dissolved in 12 ml of a 0.1 M MeOH-aq / NaOH solution (1: 1). The reaction mixture is agitated at room temperature for 2 h and concentrated under reduced pressure. The residue East separated using EtOAc and aqueous citric acid (0.1 M). The aqueous phase East extracted twice with EtOAc. The organic phases are combined, dried over NazSOa, filtered and concentrated under reduced pressure. We get 780 mg (yield 80%) of compound Q after crystallization from Et20-heptane. 1 H NMR: 8 (DMSO-dh) 4.05 (1 H, ddd, J 4.6, 10.1 and 12.6 Hz, H-5), 3.89 (1 H, s large, H-3), 3.31 (1 H, dd, J 2.8 and 10.1 Hz, H-4), 3.15 (2 x 3 H, s, 2 OCH3), 1.89 (1 H, dd, J4.6 and 12.6 Hz, H-6), 1.74-1.64 (3 H, m, 2 H-2 and H-6 '), 1.23 and 1.14 (2 x 3 H, 2 s, 2 CH3); '3C NMR
: b (DMSO-d6) 177.0 (C00), 76.0 (C-1), 73.9 (C-4), 69.0 (C-3), 63.8 (C-5), 48.1 and 48.0 (2 OCH3), 39.1 and 39.5 (C-2 and C-6), 18.6 (2 CH3).
S b) Preparation of the tripentides of sequences (S2) and (S31.
These tripeptides have the following sequences Lys-Cys (tert-butylthio) -Gly-OH (S2) Lys-Cys (tert-butylthio) -Arg-NHZ (S3) The tripeptides (S2) and (S3) are respectively neutral and charged positively in physiological medium, as opposed to the peptide (S1) described in Example 1, which is negatively charged in a physiological medium.
They are obtained according to the same protocol as that described in Example 1 for the synthesis of the peptide (S1), except that the syntheses have been made from a Rink amide-AM-PS resin (0.70 mmol / g, Senn Chemicals), at 0.2 mmole scale. The protective group of Fmoc-L-Arg-OH is a 2,2,5,7,8-pentamethylchroman-6-sulfonyl group.
c) Coupling of shikimic acid and the quinic acid derivative (compound Q) on the peptides (S21 and (S3).
0.2 mmol of each peptidyl resin (i.e. the sequence peptide on solid support), namely 0.2 mmol of sequence (S2), of a leaves, and 0.2 mmol of sequence (S3), on the other hand, are brought into contact with 123 mg (0.4 mmol, 2 eq.) Of compound Q, in the presence of BOP (177 mg, 0.4 mmol, 2 eq.) And DIEA
(140 p.1, 0.8 mmol, 4 eq.) In DMF for 1 h and at room temperature.
The peptidyl-resin is then washed with DMF (2 x 2 minutes) and CHZCIz (3 x minute). The coupling is carried out a second time.
In addition, 0.2 mmol of each peptidyl-resin are put in presence of 61 mg (0.44 mmol, 2.2 eq.) of shikimic acid, using a activation by HBTU-HOBt-DIEA [(166 mg, 0.44 mmol, 2.2 eq.), (67 mg, 0.44 mmol, 2.2 eq.), (146 ml, 0.84 mmol, 4.2 eq.) J in the NMP, for 40 minutes at temperature ambient. The peptidyl-resin is then washed with DMF (2 x 2 minutes) and of CHZCl2 (3 x 1 minute). The coupling is carried out a second time.
The peptidyl-resins are then washed with EtzO. The peptides are separated from the solid support by treatment with a TFA-Hz0- mixture iPr3SiH
95: 2.x: 2.5 (10 ml per gram of peptidyl-resin), for 1 h at temperature ambient, precipitated in a cold diethyl ether-heptane mixture, centrifuged, dissolved in water, then freeze-dried. The peptides (S2) q, (S2) s, (S3) q and (S3s), which correspond respectively to the tripeptides (S2) and (S3) carrying two remains acid quinic (peptides (S2) q and (S3) q) or two residues of shikimic acid (peptides (S2) s and (S3) s), as shown in Figures 6 and 7.
~ Peptide (S '~ q These are Na, NE-di - [(ls ", 3R, 4s °, SR) -1,3,4, x-tetrahydroxycyclo-hexane-I-carbox-1-yl] -L-lysyl- [S- (tert-butylthio)] - L-cysteinyl-glycine-amide.
We obtains 35.6 mg (24% yield) of this product in the form of a powder white, after purification by RP-HPLC [gradient: 100: 0 to 72:28 (AB) for minutes, then 72:28 to 62:38 (AB) for 30 minutes] followed by lyophilization.
1 H NMR: 8 (DZO-H20 5:95) 8.50 (1 H, d, J 6.9 Hz, Cysteinyl NH), 8.35 (1 H, t, J 5.9 Hz, Glycyl NH), 8.16 (1 H, d, J 6.9 Hz, Lysyl ° 'NH), 8.12 (1 H, d, J
6.1 Hz, Lysyl eNH), 7.30 and 7.00 (2 x 1 H, 2 s wide, NHZ), 4.52 (1 H, m, Cysteinyl aH), 4.16 (1 H, dt, J 6.9 and 5.9 Hz, Lysyl aH), 4.09-4.08 (2 H, m, 2 H-3), 3.97-3.90 (2 H, m 2 H-5), 3.85 (1 H, dd, J 5.9 and 10.4 Hz, Glycyl aH), 3.72 (1 H, dd, J 5.9 and 10.4 Hz, Glycyl aH), 3.43-3.38 (2 H, m, H-4), 3.14-3.09 (2 H, m, Lysyl sH and Cysteinyl ~ 3H), 2.94 (1 H, dd, J 8.5 and 14.0, Cysteinyl (3H), 2.05-1.6 ~ (6 H, m, 2 H-2, 2 H-6 and Lysyl [3H), 1.45-1.38 (1 H, m, Lysyl 8H), 1.23-1.19 (2 H, m, Lysyl yH), 1.20 (3 x 3 H, s, 3 CH3);
NMR 13C: s (DZO-Hz0 5:95) 17î.5, 177.2, 174.7, 174.3 and 172.8 (~ COIS, 77.1 (2 VS-1), 75. ~ and 75.4 (2 C-4), 70.8 (2 C-3), 66.8 and 66.7 (2 C-5), 54.4 (Lysyl aC), 53.5 (Cysteinyl aC), 48.6 [C (CH3);], 42.8 (Glycyl aC), 40.7 (2 C-6), 40.3 (Cysteinyl (3C), 39.4 4 (Lysyl sC), 37.5 (2 C-2), 30.8 (Lysyl (3C), 29.4 [C (CHz);], 28.3 (Lysyl 8C), 22.7 (Lysyl yC); ml ~ (TOF-PDMS) 765 (M + Na) +.
~ Peptide (S? Js These are l ~ '~, lV ~ di - [(3R.4S.SR) -3,4, x-trihydroxy-1-cyclohexene-carbox-1-yl] -L-lysyl- [S- (tert-but <~ lthio)] - L-cysteinyl-glycine-amide. We gets 56 mg (40% yield) of this product in the form of a white powder, after purification by RP-HPLC [gradient: 100: 0 to 90:10 (AB) for 10 minutes, then 90:10 to 85:15 (A / B) for 15 minutes, then isocratic gradient (A / B)]
followed by lyophilization. 1 H-NMR: b (D20-H20 5:95) 8.46 (1 H, d, J 7.1 Hz, Cysteinyl NH), 8.27 (1 H, t, J 5.8 Hz, Glycyl NH), 8.08 (1 H, d, J 6.2 Hz, Lysyl "NH), 7.87 (1 H, t, J
~. ~ Hz, Lysyl ENH), 7.26 and 7.0 (2 x 1 H, broad s, NHZ), 6.28 and 6.22 (2 x 1 H, 2 d large, J 3.7 Hz, 2 H-2), 4.50 (1 H, m, Cysteinyl aH), 4.26 (2 H, m, 2 H-3), 4.17 (1 H, dt, J 6.2 and 6.4, Lysyl aH), 3.89-3.83 (2 H, m, 2 H-5), 3.80 and 3.76 (2 x 1 H, 2 dd, J 5.8 and 15.4 Hz, 2 Glycyl aH), 3.71-3.54 (2 H, m, 2 H-4), 3.12-3.06 (2 H, m, Lysyl EH and Cysteinyl [3H), 2.90 (1 H, dd, J 8.9 and 14.0 Hz, Cysteinyl ~ 3H), 2.62-2.53 (2 H, m, 2 H-6), 2.05 (2 H, dd, J6.9 and 18.7 Hz, 2 H-6 '), 1.76-1.56 (2 H, m, 2 Lysyl [iH), 1.43-1.37 (2 H, m. 2 Lysyl 8H), 1.27-1.15 (2 H, m, 2 Lysyl yH), 1.15 (3 x 3 H, s, 3 CH3); KIDNEY
~ 3C: S (Dz0-H ~ 0 5:95) 175.0, 174.4, 172.9, 171.0 and 170.5 (~ CON), 133.7 and 133.0 (2 C-2), 132.2 and 131.0 (2 C-1), 72.0 and 71.9 (2 C-3), 66.8 and 66.3 (2 C-4 and 2 C-5), 55.0 (Lysyl aC), 53.7 (Cysteinyl aC), 48.7 [C (CH3) 3J, 42.7 (Glycyl acC), 40.2 (Cysteinyl ~ C), 39.8 (Lysyl $ C), 31.7 and 31.5 (2 C-6), 30.6 {Lysyl ~ 3C), 29.4 [C (CH3) 3J, 28.3 (Lysyl SC), 22.9 (Lysyl yC); m / z (TOF-PDMS) 729 (M + Na) +.
~ Peptide (S3) q These are Na, l ~ di - [(ls ", 3R, 4s °, SR) -1,3,4,5-tetrahydroxycyclo-hexane-1-carbox-1-yl] -L-lysyl- [S- (tert-butylthio)] - L-cysteinyl-L-arginine-amide. We obtains 45 mg (24% yield) of this product in the form of a powder white, after purification by RP-HPLC [gradient: 100: 0 to 85:15 (A / B) for 15 minutes, then 85:15 to 75:25 (A / B) for 30 minutes) followed by lyophilization. NMR
1H: S
(D20-H ~ 0 5:95) 8.39 (1 H, d, J7.0 Hz, Cysteinyl NH), 8.27 (1 H, d, J7.5 Hz, Lysyl "NH), 8.14 (1 H, d, J 6.9 Hz, Arginyl" NH), 8.12 (1 H, t, J 6.1 Hz, Lysyl ~ NI ~, 7.42 and 7.0 (2 H. 2 s, NHZ), 7.1 (1 H, t, J5.3 Hz, Arginyl iVri), 6.55 (s large, guanidinium), 4.50 (1 H, m, Cysteinyl aH), 4.30-4.10 (2 H, m, Arginyl aH and Lysyl aH), 4.09 (2 Hrs, s large, 2 H-3), 3.98-3.89 (2 H, m, 2 H-5), 3.43-3.37 (2 H, m, 2 H-4), 3.10-3.05 (5 H, m, 2 Arginyl 8H, 2 Lysyl sH and Cysteinyl ~ 3H), 2.97 (1H, dd, J 8.9 and 14.0 Hz, Cysteinyl ~ 3H), 1.95-1.54 (8 H, m, 2 Arginyl [iH, 2 Lysyl ~ iH, 2 H-2 and 2 H-6), 1.55-1.48 (2 H, m, 2: lrginyl yH), 1.49-1.32 (2 H, m, 2 Lysyl 8H), 1.27-1.15 (2 H, m, 2 Lysyl yH), 1.20 (3 x 3 H, s, 3 CH3); 3N NMR: b (D20-H ~ O 5:95) 177.5, 177.2, 176.2, 174.5 and 172.1 (5 CON), 157.3 [NH (C = NH) NHz], 77.2 (2 C-1), 75.5 (2 C-3), 70.9 and 70.8 (2 C-5), 66.8 and 66.7 (2 C-4), 55.6 (Cysteinyl aH), 53.6 (Lysyl aH and Arginyl aH), 48.8 [C (CH3) 3], 41.0 and 40.5 6 (Cysteinyl [iH and Arginyl 8H), 40.7 (2 C-6), 39.4 (Lysyl sH), 37.5 (2 C-2), 30.9 (Lysyl (3H), 29.4 [C (CH3) 3], 28.6 and 28.4 (Lysyl 8H and Arginyl ~ 3H), 24.9 (Arginyl yH), 22.7 (Lysyl yH); m / z (TOF-PDMS) 842 (M + H) +.
~ Peptide (S3) s This is lVal ~ di - [(3R, 4S, 5R) -3,4,5-trihydroxy-1-cyclohexene-carbox-1-yl] -L-lysyl- [S- (tert-butylthio)] - L-cysteinyl-L-arginine-amide. We gets 48 mg (26% yield) of this product as a white powder, after purification by RP-HPLC [gradient: 100: 0 to 90:10 (A / B) for 10 minutes, then 90:10 to 85:15 (A / B) for 12 minutes, then isocratic gradient] followed of a lyophilization. 1 H NMR: 8 (DZO-Hz0 5:95) 8.40 (1 H, d, J 6.8 Hz, Cysteinyl NH), 8.14 (1 H, d, J 7. 8 Hz, Lysyl ° 'NH), 8.11 (1 H, d, J 7.7 Hz, Arginyl ° 'NH), 7.92 (1 H, t, J 5.7 Hz, Lysyl eNH), 7.39 and 7.05 (2 x 1 H, 2 s wide, NHZ), 7.07 (1 H, t, J
5.4 Hz, Arginyl sNH), 8.40 (1 H, d, J 6.8 Hz, Cysteinyl NH), 6.55 (s wide, guanidinium), 6.36 and G.27 (2 x 1 H, 2 d wide, J3.9 Hz, 2 H-2), 4.52 (1 H, m, Cysteinyl aH), 4.33-4.28 (2 H, m, 2 H-3), 4.21-4.16 (2 H, m, Lysyl aH and Arginyl aH), 3.92-3.88 (2 H, m, 5), 3.64-3.60 (2 H, m, 2 H-4), 3.15-2.96 (6 H, m, 2 Cysteinyl (3H, 2 Lysyl sH, Arginyl 8H), 2.65-2.53 (2 H, m, 2 H-6), 2.10 (2 H, d wide, J 6.6 Hz, 2 H-6 '), 1.78-1.65 (4 H, m, 2 Lysyl (3H and 2 Arginyl (3H), 1.54-1.40 (2 H, m, 2 Lysyl 8H and Arginyl yH), 1.31 (3 x 3 H, s, 3 CH3); 13C NMR: ~ (Dz0-Hz0 5:95) 176.3, 174.9, 172.3, 171.0 and 170.5 (5 CON), 157.2 [NH (C = NH) NHZ], 133.6 and 133.0 (2 C-2), 132.4 and 131.1 (2 C-1), 72.0 (2 C-3), 66.8 and 66.2 (2 C-4 and 2 C-5), 55.2 and 54.0 (Lysyl aC
and Arginyl aC), 53.6 (Cysteinyl aC), 48.9 [C (CH3) 3], 41.0 (Cysteinyl (3C), 40.1 (Arginyl 8C), 39.7 (Lysyl sC), 31.6 (2 C-6), 30.6 (Lysyl (3C), 29.4 [C (CH3) 3], 28.5 and 28.3 (Arginyl (3C and Lysyl SC), 24.9 (Arginyl yC), 22.9 (Lysyl yC); m / z (TOF-PDMS) 805 (M + Na) +.
d) Preparation of ligand 41F (Figure 61.
6 eq. of peptide (S2) q are dissolved in a degassed mixture from nPrOH-Hz0 50:50 (1 ml). We introduce 6 eq. from nBu3P. The mixture is stirred at room temperature for 24 h, under nitrogen. The solvent is evaporated under pressure reduced and the residue obtained is further dried under vacuum for 15 minutes. We add 500 ~, l of the degassed mixture DMF-aq NaOAc 0.1 M (90:10), then 0.9 pmol (1 eq.) Of dendrimer of L-lysine 14, the preparation of which is described in Example 1.
The middle stirred for 24 h at room temperature, the progress of the reaction being followed by RP-HPLC. Fluorescein isothiocyanate (4 eq.) Is added and the medium is stirred under nitrogen and protected from light overnight. The middle east diluted in water, lyophilized and purified by RP-HPLC [gradient: 100: 0 to 85:15 (AB) for 15 minutes, then 85:15 to 80:20 (AB) for 15 minutes, then gradient isocratic] to give 2.38 mg (72% yield) of ligand 41F under the form of a white powder. His analysis is as follows: ESI-MS found 3763.0, calculated 3765.1; m / z 1883.0 (M + 2H) 2+, 1255.5 (M + 3H) 3+.
e) Preparation of ligands 42F (Figure 6), 43F and 44F (Fioure 7).
6 eq. peptide (S2) s (or peptide (S3) s, or even peptide (S3) q) are dissolved in a degassed mixture of 50:50 nPrOH-HZO (1 ml).
We introduced 6 eq. from nBu3P. The mixture is stirred at room temperature for 24 hours, under nitrogen. The solvent is evaporated under reduced pressure and the residue obtained is still dried under vacuum for 15 minutes, then dissolved in 50 p.1 of water. 1 eq. (0.5-1 p, mol) L-lysine 14 dendrimer, the preparation of which is described in the example l in 300 p, 1 of DMF comprising 4 eq. of DIEA is added to 1 eq. isothiocyanate fluorescein. The mixture is stirred under nitrogen and protected from light for 2 h. A
once the reaction is complete (according to its monitoring by RP-HPLC), the mixture is added to reaction medium containing the peptide compounds. The pH of the mixture is adjusted to 8-8.5 by addition of solid KZC03. The mixture is stirred under nitrogen and under cover of the light for 24 h, at room temperature, diluted in water, freeze-dried and purified by RP-HPLC to give ligands 42F, 43F or 44F.
1.3 mg (36% yield) of ligand 42F are obtained in the form of a white powder, after purification by RP-HPLC HPLC [gradient: 100: 0 to 82:18 (AB) for 18 minutes, then 82:18 to 72:28 (AB) for 30 minutes], followed by lyophilization. His analysis is as follows: ESI-MS found 3620.0, calculated 3621.0; m / z 1207.7 (M + 3H) 3+, 1001.8 (M + 3H-CZSH3gN50 ") 3+, 905.9 (M + 4H) 4+.
1.3 mg (36% yield) of ligand 43F are obtained in the form of a white powder, after purification by RP-HPLC HPLC [gradient: 100: 0 to 82:18 (A / B) for 18 minutes, then 82:18 to 72:28 (A / B) for 30 minutes], followed by lyophilization. His analysis is as follows: ESI-MS found 4160.0, calculated 4161.7; m / z 1387.6 (M + 3H) 3+, 1041.0 (M + 4H) 4+, 833.0 (M + SH) s +.
1.3 mg (36% yield) of ligand 44F are obtained in the form of a white powder, after purification by RP-HPLC HPLC [gradient: 100: 0 to 82:18 (A / B) for 18 minutes, then 82:18 to 75:25 (A / B) for 21 minutes, then isocratic gradient], followed by lyophilization. His analysis is the next: ESI-MS
found 4016, calculated 4017.6; m / z 1339.9 (M + 3H) 3+, 1005.2 (M + 4H) 4+, 804.4 (M + SH) s +.
EXAMPLE 6. PREPARATION OF LIGANDS CONFORMING TO
THE INVENTION
Two ligands corresponding to the general formula (III) in accordance with the invention, in which m is an integer equal to 4 or 8, ~ n is an integer equal to 0 (and therefore X is absent), . y is an integer equal to 1, . B ~ represents an acid (-) - quinic residue, . Y represents the rest of an antigen, ~ A represents the remainder of a dendrimer A 'comprising 8 (in the case of the ligand 51 shown in Figure 10) or 4 (in the case of ligand 52 shown in the Figure 11) L-lysine residues, ~ M and P respectively represent thioether groups (-CHz-S-CHZ-CO-) and hydrazone (-CO-CH = N-NH-CHZ-CO-), were prepared according to the following protocol.
These ligands are obtained by ligation of derivatives of quinic acid, namely quinic acid functionalized by a thiol function (compound G), a lysine dendrimer (H or I dendrimer) and a peptide antigen (antigen J or K), according to the protocol described below.

a) Preparation of the q-uinic acid derivative (compound G Fieure 8).
Compound G represents 2- {N, N di - [(ls ", 3R, 4s ~, 5R) - disulfide -1,3,4,5-tetrahydroxycyclohexane-1-carboxamido-1-yl]} ethyl. It is obtained in mixing 52 mg (0.3 mmol) of acid overnight at 50 ° C
quinic lactone and S 34 mg (1 eq.) Cysteamine hydrochloride, in degassed water containing of Solid KZC03 (pH 8-9). The mixture is then stirred at room temperature while 24 h, in the presence of air, before being purified by RP-HPLC [gradient: 100: 0 to 80:20 (A / C) for 15 minutes, then isocratic], then lyophilized to give the compound G (70 mg, 71%) in the form of a white powder. His analysis is as follows 1 H NMR: 8 = 8.24 (t, 3J (H, H) = 5.8 Hz, 2 H; 2 NH), 4.05 (ddd, 3J (H, H) = 3, 3 and 6.2 Hz, 2H; 2 H-3), 3.90 (ddd, 3J (H, H) = 4.7, 9 and 11.6 Hz, 2 H; 2 HS), 3.40 (t, 3J (H, H) _ 6.4 Hz, 4, H; 2 CHZ), 3.38 (dd, 3J (H, H) = 3 and 9 Hz, 2 H; 2 H-4), 2.73 (t, 3J (H, H) = 6.4 Hz, 4H; 2 CHZ), 1.94-1.83 (m, 6 H; 4 H-2 and 2 H-6), 1.77 (dd, 3J (H, H) = 11.6 and 13.4 Hz, 2H; 2 H-6 '); 13C NMR: b = 177.5 (2 COIS, 77.8 (2 C-1), 75.4 (2 C-4), 70.8 (2 C-3), 66.7 (2 C-5), 40.7 (2 C-6), 38.5, 37.5 and 37.1 (4 CHZ and 2 C-2); TOF-PDMS:
m / z 469 [M + H] +.
b) Preparation of dendrimers H (Fig_ure 81 and I (Fieure 91.
~ Functionalization of an amino-PEGA resin by an arm spacer.
Is added successively to a solution of dimethyl ester of (+) - 2,3-O-isopropylidene-D-tartaric acid (733 p1), 7.2 p1 (4 eq.) of water and 59.6 ~ l (4 eq.) Of 1,8-diazabicyclo [5,4,0] undéc-7-ene, at room temperature. The middle reaction mixture is stirred for 1 h, then added to 0.1 mmol of an amino resin PEGA
(0.4 mmol / g) soaked in a minimum amount of DMF, the resin having, at previously, was neutralized by 5% DIEA in CHZCIz (2 x 1 minute) and DMF
(3 x 1 minute). BOP (177 mg, 4 eq.) Is then added to DMF (1 ml) and the mixture obtained is stirred at room temperature for 40 minutes. Resin East washed with DMF (4 x 2 minutes) and CHZCIz (2 x Z minutes). She is soaked with DMF and acylated with 1,3-diaminopropane (0.65 ml) at room temperature while 1 hr. Finally, the resin is washed with DMF (3 x 1 minute) and CHZCl2 (2 x 2 minutes).

~ Synthesis of L-lysine-based dendrimers.
These dendrimers are synthesized on the amino-PEGA resin functionalized as indicated above (0.2 mmol / g, Senn Chemicals) in using an Fmoc / tert-Butyle strategy, on a scale of 0.1 mmol. The stages of coupling are performed using an excess of 10 (for the first two couplings) or from 4 eq.
in amino acids with respect to reaction amino groups, activation being performed by the HBTU-HOBt-DIEA mixture in the NMP. The couplings are followed, for example, by the TNBS test, as described in Example 4.
The Fmoc- (3-Ala-OH is first coupled to the resin. The residues lysines are then introduced: the first lysine introduced is protected by a Boc group, the other lysines being added in the form of their Fmoc- derivatives L-Lys (Fmoc) -OH. After the fourth and fifth coupling steps, the peptides are deprotected in solid phase and acylated, in DMF, using an excess (4 eq.) chloroacetic anhydride, prepared using DCC activation. The peptidyl-resin is washed with DMF (3 x 1 minute), with CHZCIZ (3 x 2 minutes) and by EtzO
(2 x 1 minute), then dried. The Boc groups of the E-NHZ of the first residue lysine and isopropylidene group of the spacer arm fixed on the support are eliminated by a treatment with a TFA-HZO-anisole 95: 2.5: 2.5 mixture (10 ml), for 2 h at ambient temperature. The peptidyl-resin is washed with CHZC12 (5 x 1 minute) and by EtaO (2 x 1 minute), then dried and soaked in aqueous AcOH. We add NaI04 (128.3 mg, 6 eq.) Dissolved in an aqueous AcOH (33%) / H2O 1: 1 (2 ml) mixture. The mixture is stirred for 5 minutes, then 200 μl of ethylene glycol are there added. The resin is washed with water (2 x 6 ml) and the filtrates recovered are purified by RP-HPLC.
After purification by RP-HPLC [gradient: 100: 0 to 50:50 (A / B) for 120 minutes] and lyophilization, 28 mg is obtained (yield of 31%) of dendrimer H in the form of a white powder. His analysis is as follows:
m / z (ESI-MS positive) 1038.4 (M + H20) +, 1020.3 (M + H) +.
After purification by RP-HPLC [gradient: 100: 0 to 90:10 (A / B) for 10 minutes, then 90:10 to 70:30 (A / B) for 40 minutes, then isocratic] and lyophilization, 45 mg (23.5% yield) of dendrimer I is obtained under the form of a white powder. Its analysis is as follows: m / z (ESI-MS positive) 1855.5 (M + H2O) +, 1838.3 (M + H) +, 919.5 (M + 2H) 2+.

c) Preparation of the peptide antigens J and K (Figure 81.
Epitopes (peptide sequences called HA3o ~ -3 ~ 9 and TTg3o-sab in Figure 8) are synthesized on a 0.25 mmol scale using a resin Rink amide-Nleu-AM-PS (0.38 mmol / g, Senn Chemicals), on a synthesizer of Applied Biosystems A431 peptides. The protective group of side chains of Fmoc-L-Lys-OH is the group Boc; the group protective for Fmoc-L-Ser-OH, Fmoc-L-Glu-OH, Fmoc-L-Thr-OH, Fmoc-L-Tyr-OH and Fmoc-L-Asp-OH are tert-butyl; the protective group for Fmoc-L-Asn-OH, Fmoc-L-Gln-OH and Fmoc-L-His-OH is the trityl group; the protective group for Fmoc-L-Arg-OH is the 2,2,4,6,7-pentamethyldihydrobenzofuran-5- group sulfonyl.
~ Preparation of J antigen (HZN-Gly-HA3o ~ -319).
Once the peptide synthesis has been carried out, the peptidyl-resin (0.125 mmol) is unprotected and coupled twice manually using a excess from 4 eq. bromoacetic anhydride, prepared by DIC activation (during minutes) in DMF, followed by washing with DMF (3 x 1 minute) and CHZCIz (3 x 2 minutes). 66 mg (4 eq.) Of tert-butylcarbazate are added to the peptidyl-resin soaked in DMF containing DIEA (174 p, 18 eq.). The mixture is stirred a night, then washed with DMF (3 x 1 minute), CHZC12 (3 x 2 minutes) and with EtzO (2 x 1 minutes). Finally, the peptide is separated from the support and deprotected using the mixed TFA-anisole-H2O 95: 2.5: 2.5 (10 ml), for 3 h at room temperature, precipitate in cold diethyl ether, centrifuged, dissolved in water and lyophilized. We obtains 101 mg (40% yield) of J antigen in the form of a white powder after purification by semi-preparative RP-HPLC [gradient: 100: 0 to 50:50 (A / F) during 90 minutes] and freeze-drying. ESI-MS positive analysis of J antigen is the following: m / z 1575 (M + H) +.
~ Preparation of K antigen (HZN-Gly-TTs3o-sab).
We proceed as indicated above in relation to the preparation of the J antigen, except that - the peptide is separated from the support and deprotected by means of the mixture TFA-iPr3SiH-H20 95: 2.5: 2.5 (10 ml), for 1.5 h, - the gradient, for the purification of the antigen by RP-HPLC, is the next: 100: 0 to 75:25 (A / F) for 25 minutes, then isocratic (A / F).

WO 01/09173 $ 3 PCT / FR00 / 02194 After purification by RP-HPLC and lyophilization, the K antigen is obtained in the form of a white powder, with a yield of 15%. His analysis by positive ESI-MS is as follows: calculated 2052.5, found 2052.0; m / z:
1026.7 [M + 2H] 2+, 684.8 [M + 3H] 3+, 514.0 [M + 4H] 4+.
d) Preparation of ligands 51 and 52.
~ Ligand 51 (Figure 10).
Compound G (1.5 eq. / Chloroacetic groups to be substituted), of which the preparation is described in paragraph a) above, is dissolved in the mixed nPrOH-Hz0 1: 1 (500 p, 1). Tri-n-butylphosphine (1 eq.) Is added. After one night at room temperature under a nitrogen atmosphere, the mixture is concentrated under reduced pressure for 20 minutes, taken up in 500 p1 of water and added to 0.8 p, mol (1 eq.) Of dendrimer I dissolved in 600 ~ l of DMF. The pH is adjusted to 8-8.5 through addition of solid carbonate potassium. The mixture is stirred for 24 to 48 h at room temperature and under a nitrogen atmosphere. Then add 350 p.1 of a citrate / phosphate buffer and peptide antigen J. The pH is adjusted to 5.2 through addition of 1N aqueous HCl. The mixture is stirred for 12 h at temperature ambient, diluted in water, frozen and lyophilized. 1.52 mg of ligand 51 (yield of 35%) are obtained, in the form of a white powder, after purification by RP-HPLC
[gradient: 100: 0 to 90:10 (A / B) for 10 minutes, then 90:10 to 75:25 (A / B) while 50 minutes] and lyophilization. Its positive ESI-MS analysis is as follows calculated 5114.1, found 5112; m / z: 1704.9 [M + 3H] 3+, 1278.9 [M + 4H] 4+, 1030.9 [M + SH + K] 5+, 859.3 [M + SH + K] 6+.
~ Ligand 52 (Figure 11).
Compound G (1.5 eq. / Chloroacetic groups to be substituted) is dissolved in the nPrOH-H2O 1: 1 mixture (500 p, 1). We add tri-n-butylphosphine (1 eq.). After overnight at room temperature under a nitrogen atmosphere, the mixed is concentrated under reduced pressure for 20 minutes. In parallel, 0.4 mol of dendrimer H are placed in the presence of 1 eq. of K antigen in the 'BuOH- mixture HZO (180-20 u1), at room temperature and under a nitrogen atmosphere. Once the reaction complete (followed by RP-HPLC), the reaction medium is added to compound G prepared above and the pH is adjusted to 8-8.5 by addition of potassium carbonate WO 01/09173 $ 4 PCT / FR00 / 02194 solid. Ligand 52 is then isolated as described above for the ligand 51.
0.71 mg of ligand 52 (42% yield) are obtained, in the form of a powder white, after purification by RP-HPLC [gradient: 100: 0 to 75:25 (A / B) for minutes, then 75:25 to 60:40 (A / B) for 30 minutes] and lyophilization. His analysis by positive ESI-MS is as follows: calculated 3913.6, found 3912; m / z:
1305.0 [M + 3H] 3+, 979.1 [M + 4H] 4+, 783.6 [M + SH] 5+.

INTERMEDIATE COMPOUNDS FOR THE PREPARATION OF LIGANDS
CONFORM TO THE INVENTION
Two compounds corresponding to the general formula (VI) in accordance with the invention, in which m is an integer equal to approximately 58 (for the compound called "5% QuinPEI" in the following) and approximately 116 (for the compound referred to as “10% QuinPEI” in the following), Bl represents an acid (-) - quinic residue, M represents an amide function (-CO-NH-), A represents the remainder of a linear polyethyleneimine (PET) polymer, were prepared by condensation of PEI on 1,5-lactone quinic acid (y-lactone resulting from the esterification between the carboxyl group carried by the carbon located in position 1 and the hydroxyl group located in position 5 of the acid (-) -quinique), according to following protocol.
D (-) - quinic acid (Aldrich) is transformed into I, 5-y-lactone bicyclic (also called "quinide") by heating in toluene, in presence p-toluenesulfonic acid, as described in particular by HOL FISCHER in Chem: Ber., 1921, 54, 775-785, by M. PHILIPPE et al. in J. Antibiotics, 1982, 25, 1507-1512 and by PQ: HUANG et al. in Tetrahedron Lett., 1995, 36, 8299-8302.
Lactone (10.1 mg, 58.1 pmol) reacts with 50 mg of PEI (25 kDa, Aldrich), this who corresponds to 1162 mol of amino groups, assuming a molecular mass weight average of 43 Da for the repetitive unit of the PEI. The reaction is performed in 2 ml of water, for 24 h and at 50 ° C. The reaction medium is then diluted and freeze-dried. PEI is obtained substituted, at a rate of 5%, with acid quinique (hereinafter referred to as "5% QuinPEr '). Using twice as much lactone (20.2 mg, 116.2 pmol), PEI is obtained substituted, at a rate of 10%, with acid quinique (hereinafter referred to as "10% QuinPEr ').
The completion of the reaction is indicated, by 1 H and 13 C NMR analysis.
of the reaction medium, by the absence of the signal corresponding to the quinide (the condensation of the quinide on the PEI in fact results in the opening of the cycle of lactone). The results are confirmed by quantification of the quantity acid quinic coupled to PEI, performed by 1 H NMR spectroscopy on samples products obtained, purified by ultrafiltration on amicon membranes ~
(30000 Da, Millipore). The quinic acid content of the PEI is confirmed by comparing the CHZ proton peak surfaces of the polymer backbone and protons H-4, H-5 and H-6 quinic acid residues.
PEI is a branched polymer with secondary amines and primary. Study of the 1 H NMR spectrum makes it possible to distinguish the acids quiniques linked via a secondary or tertiary amide bond (the latter being marked with a index '). Thus, 5% QuinPEI and 10% Quin PEI contain 30 and 26%
quinic acid residues connected via a tertiary amide link, the rest being secondary amide bonds.
In the following data (1 H and 13 C NMR analyzes carried out in DZO-H20 10:90), the internal reference is the sodium salt of the acid 3- (trimethylsilyl) [2,2,3,3-d4J propionic.
5% QuinPEI: RMN ~ HS 4.06 (wide, H-3 '), 3.98 (ddd, J = 1.1, 3.3 and 7 Hz, H-3), 3.95-3.85 (m, 1 H, H-5 '), 3.85 (ddd, J 4.8, 9.1 and 10.5 Hz, H-5), 3.39 (dd, J = 1.1 and 9.1 Hz, H-4 and H-4 '), 3.20, 3.06, 2.71, 2.60 and 2.51 (5 s large, CH and CHZ PEI), 1.80-1.68 (m, 4 H, H-2, H-2 ', H-6 and H-6'); NMR 13C 8 180.7 (CON '), 177.5 (CON), 77.4 (C-1), 77.1 (C-l '), 75.7 (C-4), 75.5 (C-4'), 70.9 (C-3 and C-3 '), 67.5 (C-5), 66.7 (C-S '), 56.5-46.0 (C PEI), 41.1 and 40.8 (C-6 and C-6'), 39.7-37.0 (C PEI), 38.0 (C-2 and C-2 ').
10% QuinPEI: NMR ~ H 8 4.05 (large d, H-3 '), 3.96 (ddd, J = 3.2, 3.2 and 7 Hz, H-3), 3.95-3.85 (m, H-5 '), 3.85 (ddd, J 4.8, 9.1 and 10.8 Hz, H-5), 3.37 (dd, J = 3.2 and 9.1 Hz, H-4 and H-4 '), 3.17, 3.00, 2.65, 2.57 and 2.49 (5 s wide, CH
and CHZ
PEI), 1.72-1.69 (m, 4 H, H-2, H-2 ', H-6 and H-6'); NMR 13C b 181.7 (CON '), 177.3 (CON), 77.4 (C-1), 77.1 (C-l '), 75.7 (C-4), 75.5 (C-4'), 70.9 (C-3 and C-3 '), 67.5 (C-5), 66.7 (C-5 '), 55.4-46.0 (C PEI), 41.0 (C-6 and C-6'), 40.0-38.0 (C PEI), 37.9 (C-2 and C-2 ').

THE INVENTION
The biological activity of the ligands in accordance with the invention has been verified by a series of in vitro experiments aimed at testing their aptitude to be internalized, via the mannose receptor, by dendritic cells obtained at from human peripheral blood.
The experiments reported here were carried out with following compounds ~ methyl quinate (i.e. the methyl ester of quinic acid), which East described by E. DELFOURNE et al. in J. Chem. Res., 1991, 56-57, methyl-aD-mannopyranoside, sold by the company LANCASTER, ligands linked to fluorescein 23F and 24F, the preparation of which is described in example 2 above, ~ a compound, which will be called compound 30F in what follows and is differentiating ligands 23F and 24F in that it comprises 8 chains polyhydroxylated galacto configuration instead of the 8 acid residues (-) -shikimic or (-) - quinic, this compound being intended to serve as a negative "control" in Reason to the low affinity of the mannose receptor for D-galactose and compounds related to the latter, and ~ a compound, which will be referred to as compound 31F in the following and which occurs in the form of a dendrimer consisting of 8 L-lysine residues and 8 residues D-mannose, this compound being intended to serve as a positive "control" since the mannose receptor binds preferentially to D-mannose.
1) - Protocol a) Preparation of compounds 30F and 31F.
Compound 30F was prepared according to an operating protocol consistent ~ firstly, to synthesize a tripeptide of sequence (S1) in conditions similar to those described in ~ 1.2 of (example 1 above for the preparation of compound 1;
. then, after cleavage of this tripeptide, by treatment of the resin with 25 ml of a TFA / H2O / MeZS mixture (95: 2.5: 2.5) at room temperature for 2 hours, coupling 1 molecule of said tripeptide with 2 molecules of D-galactonolactone, in reacting 160 mg (0.90 mmol) of D-galactonolactone with a solution containing 140 mg (0.22 mmol) of tripeptide and 0.79 mmol of DIEA in 5 ml methanol brought to reflux for 24 hours, then adding to this mixture reaction 80 mg (0.45 mmol) of additional D-galactonolactone and maintaining reflux for another 24 hours; and ~ e ~ n, by coupling, under conditions identical to those described in Example 2 above, N ", N ~ di - [(2R, 3R, 4S, SR) -2,3,4,5-tetrahydroxy-1-hexane-carbox-1-yl] -L-lysyl- {S- (tert-butylthio) JL-cysteyl-glycine thus obtained with a compound 14 having previously reacted with FITC.
Compound 31F was, in turn, prepared by treating, a compound 15 with 2-thioethyl-oc, D-mannopyranoside, then reacting the compound resulting with FITC.
b) Obtaining dendritic cells.
Dendritic cells were obtained by differentiation of monocytes, themselves isolated from mononuclear blood cells peripheral human, according to a protocol derived from that described by AVRAMEAS et al. (Eur. J.
Immunol., 1996, 26, 394-400).
To do this, the mononuclear cells present in human peripheral blood samples (supplied by the establishment of Transfusion Sanguine from Nord-Pas de Calais, FRANCE) have been separated from the other elements of blood by Ficoll / Passover density gradient centrifugation (Company PHARMACIA), then washed 3 times, by centrifugation, with RPMI 1640 medium (Company GIBCO) containing 3 mM EDTA.
The mononuclear cells thus isolated were suspended, at 37 ° C and under an atmosphere with 5% COZ, in a culture medium including RPMI 1640 medium supplemented with 5.10-5 M of (i-mercaptoethanol (Company MERCK), 2 mM L-glutamine (MERCK Company), 1 mM sodium pyruvate (GIBCO company), 10% fetal calf serum inactivated by heat (Company GIBCO), 100 LJI / ml of penicillin and per 100 ~ g / ml of streptomycin (all of them from SPECIA).
Then, the resulting suspension was distributed over Petri dishes of cm in diameter, at a rate of 8 to 12.10 'cells per disc. Enrichment in monocytes was obtained by letting these cells adhere to the dishes, to 37 ° C and in a humidified COZ (5%) incubator for 3 to 4 hours.
10 After elimination of the non-adherent cells, the cells adherents were returned to culture in the medium described above, but including in addition, 800 U / ml of growth factor from granulocytic lines and recombinant human macrophage (PEPRO TECH company) and 1000 U / ml of recombinant human interleukin-4 (Company PEPRO TECH), at a rate of 106 cells per ml of medium. They were then cultivated, in plates of cell culture of 24 wells and under the same temperature conditions and atmosphere than before, for 6 to 8 days during which there is differentiation of monocytes towards dendritic cells (the latter being weakly adherent).
c) Analysis by flow cytometry of non-adherent cells collected.
In order to verify that the non-adherent cells collected at (step b) above clearly present the surface antigens characteristic of cells dendritics, part of these cells were washed twice with buffer PBS
containing 0.03 mg / ml bovine serum albumin, then incubated, on ice cream and for 30 minutes, with a mouse monoclonal antibody directed against the human mannose receptor and conjugated to phycoerythrin (BECTON Company DICKINSON) and ~ or a mouse monoclonal antibody directed against (human CDIa antigen, which is one of the main cell surface antigens dendritics, ~ or a mouse monoclonal antibody directed against (human CD14 antigen, which represents one of the surface antigens specific to monocytes and macrophages, these last two antibodies being labeled with FITC (BECTON Company DICKINSON).
A (from these incubations, the cells were washed with PBS buffer and resuspended in this same buffer. Fluorescence their being associated was then analyzed by cytofluorometry, using a cytometer EPICS XL-MCL (COLTLTER CORPORATION).
d) Test for inhibition of the internalization of compound 31F by the methvl-quinate.
To show that methyl quinate is a mimic of mannose in internalization with the mannose receptor of dendritic cells, the competition between methyl quinate, or methyl-aD-mannopyranoside (serving of inhibition control), and compound 31F is studied according to the protocol following the dendritic cells are recovered, washed with PBS buffer, preincubated at 37 ° C, then incubated for 20 minutes, at 37 ° C, with concentration solutions increasing in methyl quinate and compound 31F (in final concentration of SpM).
The cells are then washed with PBS, fixed in paraformaldehyde at 1%, then analyzed by flow cytofluorometry, as described in c) above.
e) Internalization of the ligands conforming to (Invention by cells dendritics.
Tests to assess the feminization of ligands 23F and 24F
were carried out according to the following protocol: non-adherent cells collected at (step b) above were washed twice with PBS buffer containing 1 mM of calcium chloride. Following which, these cells were preincubated, at 37 ° C and for 10 minutes, then incubated at 37 ° C and for 20 minutes, with different solutions each containing (one of the compounds to be tested, at a concentration between 1 and 10 pM. The cells were then washed 3 time with cold PBS buffer and fixed for 20 minutes in paraformaldehyde at 2%.
The fluorescence associated with these cells was then assessed by a FACScan analysis using the aforementioned cytometer.
Furthermore, in order to verify that the internalization of the ligands 23F and 24F by dendritic cells is well linked to recognition of these ligands by the mannose receptor and not some other mechanism, these tests were renewed in preincubating the cells in the presence of mannan solutions extracted from Saccharomyces cerevisiae (SIGMA company), mannan being, in fact, known for be selectively internalized via the mannose receptor. In this goal, the dendritic cells are recovered, washed with PBS buffer, incubated at 37 ° C, then preincubated for 20 minutes, at 37 ° C, in the presence of solutions of mannan extract of Saccharomyces cerevisiae of concentrations between 10 '~ and mg / ml. Then, ligands 23F and 24F (in final concentration of SpM) are 10 added (incubation: 20 minutes at 37 ° C). The cells are washed PBS, fixed in 1% paraformaldehyde and analyzed by flow cytofluorometry, such as described in c) above.
f) Influence of the number of mimes of mannose ~ ortés nar les ligands in accordance with the invention on the internalization of these ligands.
In order to study what is the optimal construction, i.e. what is the ligand with the lowest number of mannose mimes (for facilitate chemical synthesis of the ligand) which is specifically internalized by the mannose receptors, dendritic cells are incubated for 20 minutes to 37 ° C with 10 μM of ligands comprising 2, 4 or 8 residues of mannose, acid quinic or shikimic acid (di-, tetra- or octavalent ligands). After fixation in paraformaldehyde, the cells are analyzed by cytofluorometry of flux, as described in c) above.
For these experiments, the following ligands were used . ligands carrying shikimic acid residues: 34F (ligand divalent), 21F (tetravalent ligand) and 23F (octavalent ligand), ligands carrying quinic acid residues: 33F (ligand divalent), 22F (tetravalent ligand) and 24F (octavalent ligand).
2) - Results a) Inhibition test for the internalization of compound 31F by the methyl quinate.
Figure 12 illustrates the ability of methyl quinate to inhibit internalization of compound 31F. The percentages are shown on the ordinate inhibition, calculated by the formula inhibition = (MnX 31F - MnX mqui) / MnX 31F
or "MnX" represents the average value of the fluorescence intensity, . "MnX 31F" represents the signal due to the internalization of the compound 31F alone, . "MnX mqui" represents the signal due to the internalization of the compound 31F in presence of different concentrations (expressed in mM) of methyl quinate, indicated on the abscissa.
The curve indicated by the symbols - ~ - represents the inhibition of internalization of compound 31F with methyl quinate, and the curve indicated by the symbols -o- represents the inhibition of the internalization of compound 31F by the methyl-aD-mannopyranoside, compound serving as inhibition control.
In the presence of methyl quinate, effective inhibition is noted at 75% of specific internalization by the mannose receptor, comparable to her of the reference ligand, methyl-aD-mannopyranoside.
b) Internalization of the ligands in accordance with (Invention by cells dendritiaues.
Figure 13 illustrates, in the form of curves, the values average fluorescence intensities presented by cells dendritics after incubation in the presence of solutions comprising from 1 to 10 pM respectively of ligand 23F (-.-), ligand 24F (- ~ -), compound 30F (-o-) and compound 31F
(-o-).
The mean values of the intensities of fluorescence (AU) and, on the abscissa, the concentrations of the solutions.
Figure 13 shows that an incubation of dendritic cells, lasting 20 minutes and carried out in the presence of a solution dosed at 1 p, M
of ligand 23F or ligand 24F is sufficient to obtain internalization of these ligands by these cells. It also shows that this internalization is specific since compound 30F (carrying polyhydroxylated chains) does not penetrate practically not in dendritic cells, regardless of the dose of this compound used for (incubation of these cells, while compound 31F
(carrier of D-mannose remains) is very widely found in said cells dendritics.

Furthermore, Figure 14 illustrates, in the form of curves, the average values of fluorescence intensities presented by cells dendritics when these have been successively subjected to a preincubation in presence of mannan solutions of concentrations between 10 ~ and 10 mg / ml, S and then to an incubation with solutions comprising 5 μM of ligand 23F (-.-), of ligand 24F (- ~ -), of compound 30F (-o-) or of compound 31F (-o-). Are expressed, in ordinates, the mean fluorescence values (AU) and, on the abscissa, the concentrations of mannan solutions.
Figure 14 shows that internalization, by cells dendritics, ligands 23F and 24F, as well as that of compound 31F decreases drastically as the concentration of mannan increases.
This sign (existence of strong competition between these ligands and the mannan vis-à-vis live of mannose receptor and confirms that the internalization of ligands conform to the invention by dendritic cells is done well through of the receiver mannose.
c) Influence of the number of mannose mimes worn by the ligands in accordance with the Invention on the internalization of these lisands.
Figure 15 shows, on the ordinate, the percentage specific internalization of di-, tetra- or octavalent ligands (abscissa) by the mannose receptor. The tetra- or octavalent ligands are internalized from way preferential by the mannose receptor, compared to the divalent ligands.
We can therefore consider that the optimal structures sought are those including 4 mannose mimes, which are more easily synthesized than those comprising 8 mannose mimes. In contrast to the natural ligand (ligand carrying residues of mannose), whose internalisation is optimal for an octaval structure, we finds that ligands carrying quinic or shikimic acid residues are internalized optimally when they are tretravalent.

EXAMPLE 9: SPECIFICITY OF THE INTERNALIZATION OF LIGANDS
CONFORM TO THE INVENTION IN DENDRITIC CELLS
VIA MANNOSE RECEPTORS
1) Analysis by confocal microscopy.
The internalization and distribution of ligands conforming to the invention in dendric cells is analyzed by microscopy confocal with double and triple labeling of the mannose receptor and the cell nucleus.
Dendritic cells, differentiated from cells mononuclear in the presence of IL-4 and the growth factor GM-CSF, are used to D + 5. They are incubated for 45 minutes at 4 ° C with 10 p, g of friend mannose human receptor (PHARMIGEI ~ and 5 ~ .M from one of the following constructs . 61F (Figure 16): dendrimer consisting of 4 L-lysine residues bearing 4 remains of D-mannose, as well as a fluorescein molecule (FITC).
The construction 61F was prepared as compound 31F (cf. example 8, paragraph 1.a), but by reacting 2-thioethyl-a, D-mannopyrannoside with a compound 14 (cf. example 1, paragraph 1.1);
. 21F, 22F and 64F: constructions carrying remains respectively shikimic acid, quinic acid and polyhydroxy chains of configuration galacto (from galactonolactone molecules). Construction 64F has summer prepared as compound 30F (cf. example 8, paragraph 1.a), but in using a compound 13 for coupling (cf. example 1, paragraph 1.1). The preparation of ligands 21F and 22F is described in Example 2 above.
Structure 61 F serves as a positive control, while structure 64F
serves as a negative control. The 21F and 22F structures represent ligands conform to the present invention.
The cells are placed at 37 ° C for 5 minutes. After incubation, they are fixed for 45 minutes at 4 ° C with a solution of following composition: PBS (0.1 mM MgClz, O.ImM Ca2), paraformaldehyde at 4%, sucrose. The fixation reaction is stopped by means of a solution of chloride ammonium (SOmM), for 10 minutes and at room temperature. Cells are washed 3 times with PBS, then deposited on poly-L-lysine slides.
The signal fluorescein is amplified by incubation at 37 ° C for 30 minutes, with a Anti-FITC rabbit IgG coupled to Alexa Fluor 488 (dilution: 1 / 200é '"~
(MOLECULAR PROBES) at the same time as the incubation with the second antibody goat, anti-mouse IgG coupled to Alexa Fluor 568 (dilution: 1 / SOOeme) (MOLECULAR PROBES), allowing the detection of the anti-mannose receptor.
Incubation with the second antibodies is carried out in PBS, BSA medium (lmg / ml), saponin (0.05%).
The cells are washed with PBS containing 1 mg / ml of BSA and 0.05% saponin, then with PBS / BSA and finally with PBS, then mounted between blade and coverslip in the middle of VECTASHIELD mounting. The observation of the blades is carried out by confocal microscopy on a LEICA TCS NT device with a laser Krypton / Argon (excitation 488, 468 and 647}. The acquisition of the two lengths wave (namely, the acquisition of the fluorescence emitted by the Alexa 568, corresponding to mannose receptor, and the acquisition of fluorescence emitted by Alexa 488, corresponding to the structures tested) is carried out simultaneously and the overlay of the two images allows you to view the two markers at the same time, on the same cell section, made along the z axis of the cell.
We notice a co-location of the two markers, and therefore a co-localization of ligands 61F, 21F and 22F labeled with fluorescein with the mannose receptor, making it possible to affirm that the ligands conform to the invention are well internalized in dendritic cells via the receptor of the mannose.
2) Analysis using Cos cells transfected with mannose receptor.
Cos-1 cells, not expressing, in the native state, the receptor for mannose, are transfected with the plasmid CD8 containing a coding sequence for the mannose receptor (MR). 48 hours after transfection, the cells are collected: 10 to 15% of them express the transgene. In the following, these cells are called "Cos MR".
Internalization tests are carried out using these cells, using the following compounds 22F: dendrimer consisting of 4 L-lysine residues carrying 4 residues quinic acid, as well as a fluorescein molecule (FITC). It is a ligand in accordance with the present invention.

64F: construction carrying polyhydroxylated chains of galacto configuration (negative control.
Internalization tests consist in incubating the cells, during 20 minutes and at 37 ° C., in the presence of 10 p, moles of the compounds 22F or 64F, then to wash the cells with cold PBS and fix them with paraformaldehyde to 1%, before perform the FACScan analysis using the cytometer described in Example 8.
The Figure 17 represents the percentages of positive cells detected (in abscissa) for the following four tests, indicated on the ordinate axis . test a): incubation of Cos MR cells with compound 64F, . test b): incubation of Cos MR cells with compound 22F, in presence of a mannan solution (5 mg / ml) as described in the example 8, the mannan. competing with compound 22F against the receptor for mannose, . test c): incubation of Cos MR cells with compound 22F, . test d): incubation of Cos-1 cells (not transfected) with the compound 22F.
According to Figure 17, the non-transfected Cos-1 cells (test d) do not give, of course, no internalization of compound 22F: the signal detected is comparable to the background noise of the device. It's the same for the Cos MR cells incubated with compound 64F, not specific for the receptor for mannose (test a). Test b) shows the inhibition of the internalization of the compound 22F by the mannan. As for the 22F construction, it is well internalized so specific by cells transfected by the MR receptor (test c).

Claims (26)

1. Use of at least one compound corresponding to the general formula (Ia) and/or at least one compound corresponding to the general formula (Ib):
in which :
- R1, R2, R3 and R4 represent, independently of each other, an atom hydrogen or a protecting group, - R5 represents an -OH group, an -SH group, an -NH-NH2 group, an atom halogen, or uses a linear, branched or cyclic, saturated or unsaturated carbon chain, comprising of 1 to 18 carbon atoms and optionally from 1 to 12 atoms chosen from oxygen, the sulfur and nitrogen, said carbon chain being optionally substituted by 1 to 16 atoms halogen and optionally bearing at least one nucleophilic group or at minus one electrophilic group, as a ligand for the mannose receptor or any receptor of the family C-lectins related to the mannose receptor, or for the preparation of such a ligand. 2. Use according to claim 1, in which the compounds of general formulas (Ia) and/or (Ib) are such that the groups -OR2 and -OR3 are Related trans. 3. Use according to claim 1 or claim 2, wherein the compounds of general formulas (Ia) and/or (Ib) are such that R5 represents a group hydroxyl. 4. Use according to any one of the preceding claims, in which the compound of formula (Ib) is such that R1, R2 and R3 represent atoms of hydrogen and R5 represents a hydroxyl group. 5. Use according to claim 4, in which the compound of formula (Ib) represents (-)-shikimic acid. 6. Use according to any one of claims 1 to 3, in which the compound of formula (Ia) is such that R1, R1, R3 and R4 represent hydrogen atoms and R5 represents a hydroxyl group. 7. Use according to claim 6, in which the compound of formula (Ia) represents (-)-quinic acid. 8. Use according to claim 1 or claim 2, wherein the compound of formula (Ia) is such that R1, R2, R3 and R4 represent atoms hydrogen and R5 is NH-(CH2)2-SH. 9. Compound useful as a ligand for the mannose receptor or any mannose receptor related C-lectin family receptor, or inasmuch as intermediate compound for the preparation of such a ligand, characterized in that that he answers the general formula (III) below:

(B1M)m(XN)n A(PY)y (III) in which:

- B1 responds to one or more general formulas (IIa) or (IIb) below:

where W represents a bond or a linear, branched or cyclic, saturated or unsaturated, comprising from 1 to 18 carbon atoms and possibly of 1 to 12 atoms selected from oxygen, sulfur and nitrogen, said chain being carbonaceous optionally substituted by 1 to 16 halogen atoms, - X represents the remainder of an oligopeptide X' comprising from 1 to 6 acids amino, - A represents the residue of an at least difunctional compound A', - Y represents the residue of a compound of interest Y', - m is an integer between 1 and 32, - n is an integer between 0 and 32, - y is an integer between 1 and 4, and - M, N and P each represent a linking group, respectively between B1 and When n = 0 or B1 and X when n is different from 0, between X and A when n is different from 0, and between A and Y, and include, independently of the uses of the others, a function chosen from the oxime, hydrazone, amide, ester, thioester, hydrazide, hydroxemate, ether, thioether, amine, carbonate, carbamate, thiocarbonate, thiocarbamate, urea, thiourea and thiazolidine. 10. A compound according to claim 9, in which compound A' is formed from a sequence comprising several remains of a compound to the monk trifunctional, linked to each other by covalent bonds, and specific to offer, at the level of groups free of these remains, several anchorage sites for the compounds of formula general (Ia) and/or (Ib) or, when X is present, the compound X', and for the compound Y'. 11. A compound according to claim 10, in which compound A' comprises from 2 to 32 and preferably from 4 to 16 residues of a compound trifunctional. 12. A compound according to any one of claims 9 to 11, in which the compound A' is either in a linear form, or in a form branched and, in particular, of the tree type. 13. A compound according to any one of claims 9 to 12, in which compound A' comprises a sequence of amino acids, identical or different, selected from the group consisting of lysine, hydroxylysine, serine, threonine, cysteine, ornithine, aspartic acid and glutamic acid. 14. A compound according to any one of claims 9 to 13, in which the compound of general formula (III) is such that m is an integer between 4 and 16, n is an integer between 2 and 8, X represents the remainder of a oligopeptide comprising from 2 to 4 amino acid residues, and A represents the residue of a sequence comprising from 2 to 8 lysine residues and in the form of a dendrimer. 15. A compound according to any one of claims 9 to 14, wherein the compound of general formula (III) is such that B1 represents the residue of a or many compounds selected from (-)-shikimic acid and (-)-quinic acid. 16. A compound according to any one of claims 9 to 14, wherein Y' is selected from antigens, antibodies, nucleic acids and acid fragments nuclei coding for a protein of interest or for one or more determinants antigens of this protein, the bacterial and viral fractions purified, the principles drug actives and detectable markers. 17. Compound useful as an intermediate compound for the preparation of a ligand of the mannose receptor or of any receptor of the C- family lectins related to the mannose receptor, characterized in that it has the formula general (V) below:
(B2M)m(XN)n A~~(V) in which:
- B2 responds to one or more general formulas (IVa) or (IVb) below:

where R1, R2, R3, R4 and R5 are as defined in any one of claims 1 to 8, and where W is as defined in claim 9, - X represents the remainder of an oligopeptide X' comprising from 1 to 6 acids amino, - A represents the residue of a compound A' which is at least difunctional and comprises a sequence of amino acids, identical or different, selected from the formed group by lysine, hydroxylyaine, serine, threonine, cyrsteine, omithine, aspartic acid and glutamic acid, - m is an integer between 1 and 32, - n is an integer between 0 and 32, and - M and N each represent a linking group, respectively between B2 and When n =
0 or B2 and X when n is different from 0, and between X and A when n is different of 0, and include, independently of each other, a function chosen from among the functions oxime, hydrazone, amide, ester, thioester, hydrazide, hydroxamate, ether, thioether, amino, carbonate, carbamate; thiocarbonate, thiocarbamate, urea, thiourea and thiazolidine. 18. Compound of general formula (V) according to claim 17, in which M, N, m, n, as well as the compounds A' and X', are as defined in one any of claims 9 to 12 and 14, 19. A compound according to claim 17 or claim 18, wherein B2 represents a residue of one or more compounds chosen from the acid (-)ahikimic acid and (-)-quinic acid, optionally in protected form. 20. Compound useful as an intermediate compound for the preparation of a ligand of the mannose receptor or of any receptor of the C- family lectins related to the mannose receptor, characterized in that it corresponds to the formula general (IV) below:
(B1M)m A (VI) in which:
- B1 and M are as defined in claim 9 or claim 15, - m is an integer between 50 and 500, and - A represents the residue of a polymer A' capable of forming a non-covalent with a compound of interest, such as a nucleic acid. 21. Compound according to claim 20, in which the polymer A′ is a cationic polymer, such as a lysine polymer or a polyethyleneimine. 22. Compound useful as a mannose receptor ligand or any receptor of the C-lectin family related to the mannose receptor, characterized in that that it is in the form of a non-covalent complex between a compound responding to the general formula (VI) as defined in claim 20 or claim 21, and a compound of interest, such as a nucleic acid. 23. Use of at least one compound corresponding to the general formula (Ia) and/or of at least one compound corresponding to the general formula (Ib) for the preparation of a drug capable of binding to the mannose receptor or any receptor of the family of C-lectins related to the mannose receptor. 24. Compound useful as a mannose receptor ligand or any receptor of the C-lectin family related to the mannose receptor according to moon any of claims 9 to 16 and 22, for use as a medication. 25. A compound useful as a ligand for the mannose receptor or any receptor of the C-lectin family related to the mannose receptor according to moon any of claims 9 to 16 and 22, for in vitro use. 26. Diagnostic reagent, characterized in that it comprises a useful compound as a ligand for the mannose receptor or any receptor of the family C-lectins related to the mannose receptor according to any one of claims 9 at 16 and 22.