EP1817322A2

EP1817322A2 - Silanizing agents comprising a saccharide end group and uses thereof, such as for the functionalization of solid supports

Info

Publication number: EP1817322A2
Application number: EP05817494A
Authority: EP
Inventors: Véronique ROBERT; Françoise Vinet
Original assignee: Commissariat a lEnergie Atomique CEA
Current assignee: Commissariat a lEnergie Atomique et aux Energies Alternatives CEA
Priority date: 2004-11-16
Filing date: 2005-11-15
Publication date: 2007-08-15
Also published as: JP2008520965A; US20090142854A1; WO2006053972A3; WO2006053972A2

Abstract

The invention relates to silanizing agents comprising a saccharide end group and to the use thereof for the functionalization of solid supports. The invention also relates to solid supports that have been functionalized by said silanizing agents (glycochips) and to the use of same, such for biological analysis and, in particular, for screening saccharide molecules or proteinaceous ligands of interest.

Description

SILANIZING AGENTS WITH SACCHARIDE GROUPING

TERMINAL AND THEIR USES, IN PARTICULAR FOR THE

FUNCTIONALIZATION OF SOLID SUPPORTS

The present invention relates to silanizing agents comprising a terminal saccharide group and to their use for the functionalization of solid supports. The present invention also relates to the solid supports functionalized by these silanisation agents (sugar chips), as well as to their use, in particular for the biological analysis and in particular for the screening of saccharide molecules or protein ligands of interest . The development of microarray technologies has led to a significant advance in programs related to functional genomics. Indeed, the miniaturization of DNA deposition or synthesis techniques has led to the realization of parallelized analyzes, thus multi-parameterizable, of DNA on chips. More recently, the emergence of proteomics has given rise to the concept of protein chips (Zhu and Sydner, Current Op, in Chem Biol., 2003, 7, 55-63). These allow parallel analysis of protein / ligand interactions.

Even more recently, research in biology has focused on "glycomics", that is, the systematic study of carbohydrate / protein interactions. Indeed, glycoconjugates (that is to say any molecule having a glycan-type domain, such as glycoproteins, glycolipids, proteoglycans, and more generally any molecule containing carbohydrates), have a particularly broad functional repertoire. Chemically, these carbohydrates are molecules formed by the assembly of simple monomeric blocks. These assemblages can be of natural origin, and possibly fractionated, or of synthetic origin. The various functions of molecules belonging to the carbohydrate family rely on the ability of carbohydrate structures to interact with a very large number of molecules. The analysis of the mechanisms of recognition between carbohydrates and other molecules is a field of research in full emergence. In particular, it should enable the development of new therapeutic molecules and a better appreciation of the toxicological risks of certain molecules. On time Currently, there are few systematic methods for producing saccharide molecules. As a result, the determination of the structural characteristics involved in an interaction between a molecule and a carbohydrate, as well as the characterization of the interaction itself, implies undertaking long and tedious work. It is therefore necessary, in order to progress in the knowledge of the interaction mechanisms between saccharide-type molecules and their ligands, to be able to screen libraries of saccharide-type molecules with respect to a particular ligand, for example.

That is why we now see that a new type of biochip emerges: different types of sugar chips or "Glycochip", "carbohydrate array" or even "oligosaccharide array" in English, which constitute an evolution of the DNA or protein chip discussed above, have therefore been proposed by different authors.

These sugar chips are either the result of a deposit on a given substrate of a natural or synthetic saccharide substance (ex situ synthesis), or the result of a supported multiparallel synthesis (combinatorial chemistry) of different oligosaccharide sequences (synthesis in vitro). situ), representative of the molecular diversity of some large families of endogenous glycoconjugates such as heparans for example. The invention which will be described below is part of this technology being particularly well suited to the manufacture of sugar chips, including allowing the attachment of saccharide molecules on solid supports in a preparation process easier to put than the known methods of the prior art. Indeed, in the majority of biochips, a spacer arm makes the link between the surface of the solid support and the terminal functional unit characterizing the biochip: oligopeptides, oligonucleotides (Osborn HMI et al, Tetrahedron, 1999, 55, 1807-1850; Stetsenko DA et al, Bioconjugate Chemistry, 2001, 12, 576-586), or oligosaccharides (U.S. Patent No. 6,579,725). This spacer can play several roles at once: it is a binding molecule, that is to say that it makes it possible to connect the surface of the solid support with a functional molecule (probe);

it is an arm of spatial separation, that is to say that it makes it possible to move the probe molecule away from the surface of the solid support. The proximity of the solid support with the recognition sites of the target molecules by the probe molecules can indeed hinder or prevent the probe / target recognition from taking place, and thus adversely affect the fineness and analytical quality of the biochips. This is particularly true when the probe molecules are small, especially in the case of sugar chips.

Many spacer arms have been proposed to date, but these are not entirely satisfactory in practical terms because they have a number of unresolved disadvantages:

their structure often implies a severe limitation as to the choice of the chemical processes allowing them to be fixed on the surface of the solid support,

they do not all make it possible to fix probe molecules of saccharide nature,

they are generally so stable after fixation on the surface of the solid support that their cleavage to recover the biological molecule can not be done easily and may lead to the deterioration of the latter or of the support,

- The steps required for their chemical synthesis, their attachment to the surface of the solid support and their functionalization are multiple, sometimes difficult to achieve and therefore often a high cost.

US Pat. No. 6,579,725 describes in particular a spacer arm capable of fixing probe molecules of oligosaccharide nature. However, this spacer arm, although more effective than those presented in a still older prior art, does not solve all the drawbacks mentioned above at the same time. It can be noted that its length, its functionality, its responsiveness and its size can not always be generated at will.

The inventors have therefore set themselves the goal of providing new silanization agents which make it possible to overcome all the disadvantages set out above. In particular, they aimed to provide new silanization agents for functionalizing, in a single step, the surface of a solid support with molecules of saccharide nature and this in a simple, reliable, flexible method as to the nature and length of the spacer arm and finally less expensive than the methods of the prior art. It is on this occasion that the Inventors have developed what is the subject of the present invention.

The present invention therefore firstly relates to a silanization agent with a saccharide terminal function, characterized in that it corresponds to the following formula (I): A-X-B (I) in which:

A represents a probe molecule of saccharide nature;

X represents a spacer arm consisting of a carbon or heterocarbon chain comprising two ends, one of its two ends connecting, covalently, said spacer arm X to A and the other end covalently connecting said arm; spacer X to B, said chain having at least one ethylenic unsaturation between its two ends; it being understood that said chain can not contain several acetylenic unsaturations;

- B is a silanized group. The present invention thus provides a silanized saccharide molecule, which can act as a modular spacer arm, the different structures of which influence the reactivity of the arm, that is to say its chemical, electrochemical and / or steric behavior.

According to the invention, the probe molecule of saccharide nature constituting unit A of the compounds of formula (I) above may be of natural or synthetic origin, and may optionally be protected by one or more protective groups.

This probe molecule may in particular be chosen from all the saccharide molecules to be fixed on a support, for example for reasons

- analytical - or diagnostic. ^~ Elte ^~ may in particular ^" be -synthetized in order to represent a molecule or biomolecule saccharide biological interest such as a heparan sulfate for example, or for the purpose of representing a saccharide chain acting itself as a spacer between a surface and a molecule or biomolecule of biological interest.

According to an advantageous embodiment of the invention, the probe molecule of saccharide nature has a molecular weight of between 180 and 10 000 g / mol and even more preferably between 360 and 900 g / mol. It is preferably chosen from: i) monosaccharides and in particular from glucosamine, azidoglucosamine, D-ribose, D-xylose, L-arabinose, D-glucose, D-galactose, D-mannose , 2-deoxyribose, L-fucose, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, N-acetylneuraminic acid, D-glucuronic acid, acid

L-iduronic, D-sorbitol, D-mannitol, etc. ii) Oligosaccharides and in particular among sucrose, lactose, heparan sulphate fragments, saccharide fragments of heparin, chondroitin and sulphate dermatans, Lewis antigens, etc ..., iii) poly-oligosaccharides and in particular among the saccharide fractions of heparan sulphates, heparin and chondroitin, sulphate dermatans, etc., iv) glycoconjugates , and especially among heparan sulphates, heparin, chondroitin, dermatan sulphates, etc ..., v) glycoproteins such as immunoglobulin G, hyaluronic acid, etc., vi) glycolipids such as galactosyl-ceramides, gangliosides and cerebrosides, etc., viii) glycolipoproteins such as the glycoprotein G90 extracted from tissues of earthworm Eisenia foetida (Family Lumbricidae) or the glycoprotein MPB83 supplied by the laboratory "Veterinary Labora tories Agency "(VLA, Weybridge, UK).

... One, or more than one, of the hydroxyl and / or amine functions of the saccharide moieties of the probe molecule may be protected by one or more protective groups. These protecting groups are well known to those skilled in the art and are amply described in TW Greene et al, "Protective Groups in Organic Chemistry", Second Edition, A Wiley-Interscience Publication 1991.

According to an advantageous form of the present invention, these protective groups are chosen from acetyl groups; benzyl; aryl and in particular aryl groups substituted with a radical R chosen from alkyl chains having from 1 to 40 carbon atoms; 2,2,2-trichloroethyloxycarbonyl (Troc); benzyloxycarbonyl (Z); trichloroacetamidate (TCA); tert-butyloxycarbonyl (BOC) and fluoranylmethoxycarbonyl (Fmoc). According to another advantageous embodiment of the invention, one or more of the hydroxyl and / or amine functions of the saccharide entities of the probe molecule may be substituted by one or more hydrophobic groups making it possible to make the spacer arm more specific and / or more selective towards the target molecule that will be fixed on the probe molecule and / or its role during the use of the spacer arm.

In particular, it is possible to mention the case where by protecting the hydroxyl functions by protective groups such as the benzyl, acetate, benzylidene, isopropylidene, phthalimide, etc. groups, the saccharide part can be rendered more or less hydrophobic. According to yet another advantageous embodiment of the invention, the anomeric part of the saccharide entities may be functionalized, like any glycosidic donor, by a group which will preferably be chosen according to the nature of the covalent bond ensuring the attachment of the Saccharide-type probe molecule at one of the two ends of the spacer arm X. According to an advantageous embodiment of the invention, the covalent bonds ensuring the attachment of each of the ends of the chain constituting the spacer arm X to the units A and B are derived from the reaction between a chemical function initially carried by the precursor of the spacer arm X and a complementary chemical function carried, on the one hand, by the probe-A molecule and on the other hand by the group silanized B. There are, of course, a large number of couples Potential donors / acceptors well known to those skilled in the art of sugar chemistry (Khan SH et al., "Modern Methods in Carbohydrate Synthesis", Perkin Elmer, Applied Biosystems Division, Foster City, California, USA, 1996). ) and which can be used to form said covalent bond. By way of example, mention may in particular be made of the covalent bonds resulting from the reaction between a hydroxyl radical and a group chosen from halogen atoms such as chlorine, bromine, iodine or fluorine, as well as from phosphite, trichloroacetamidate, thioalkyl, phosphate, pentenyl, sulfoxide and xanthate groups.

The spacer arm X of the compounds of formula (I) according to the invention may be of variable length and structure. As has been seen above, it nevertheless still comprises at least one ethylenic unsaturation on the chain directly connecting A and B.

According to a particularly preferred embodiment of the invention, the spacer arm X represents a linear or branched C ₂ -C ₄₀ alkyl or C ₆ -C ₄₀ aryl chain, said chain comprising at least one ethylenic unsaturation and which may optionally be interrupted by one or more heteroatoms selected from oxygen, nitrogen, sulfur and silicon, and / or one or more functions such as amide, oximes and tertiary amine functions and / or optionally substituted by one or more substituents ( preferably from 1 to 10 substituents) chosen from linear or branched C ₂ -C ₂₀ alkyl or C ₆ -C ₂₀ aryl chains, said chains optionally possibly being also interrupted by one or more heteroatoms chosen from oxygen, nitrogen, sulfur and silicon.

According to the invention, the silanized group B is preferably chosen from the groups -Si (Ri) ₃ , -SiRi (R ₂ ) ₂ and -SiRiR ₂ R ₃ in which the radicals R], R ₂ and R ₃ , independently of each other, represent a halogen atom such as fluorine or chlorine, Ci-C ₄ alkoxy radical, a Ci-C ₄ alkyl, amino or an ester radical.

Of the alkoxy radicals defined for the radicals R ₁ , R ₂ and R ₃ , the methoxy and ethoxy radicals are particularly preferred and the alkyl radicals defined for the radicals R 1, R ₂ and R ₃ , the methyl and ethyl radicals are most particularly preferred.

Among the ester functions defined for the radicals R ₁ , R ₂ and R ₃ , mention may in particular be made of the acetoxy radical. As the silanized terminal unit B, there may be mentioned trimethoxysilyl, triethoxysilyl, trimethylsilyl and triethylsilyl groups.

Among the silanizing agents of formula (I) above, those in which:

A is chosen from monosaccharides, oligosaccharides and polysaccharides, and even more particularly from oligomers such as Glc-Glc-Glc; Lac-Lac or Gal-Gal-Gal-Gal-Gal in which the abbreviations GIc, Lac and GaI respectively denote glucose, lactose and galactose;

X represents a carbon chain having from 2 to 40 carbon atoms, comprising at least one ethylenic unsaturation, said chain being linear or branched, and optionally interrupted by one or more rings and / or one or more functions such as the amide functions, oximes and tertiary amines;

B represents a trimethoxysilyl or triethoxysilyl group. As a particularly preferred compound of formula (I), there may be mentioned the compounds corresponding to formulas (1-1) and (1-2) below:

(M) in which Ac represents the acetyl group.

The silanizing agents of formula (I) above can be easily prepared according to the principles of organic synthesis well known to those skilled in the art depending on the nature of the units A, X and B.

In particular, these silanizing agents can generally be prepared by a simple assembly of the units A, X and B, said units being either previously prepared or commercially available, it being understood that said units contain the chemical functions suitable for the formation of a covalent bond on the one hand between the unit A and one end of the spacer arm X and, on the other hand, between the unit B and the other end of the spacer arm X.

The reactions used are generally conventional glycosylation reactions of A on X followed by hydrosilylation reactions (of Karstedt for example) to hang B. The silanizing agents of formula (I) according to the invention can be used to the functionalization of solid supports.

The present invention therefore relates to the use of at least one silanizing agent of formula (I) as defined above, for the functionalization of solid supports, and in particular for the manufacture of sugar chips. The use of silanization agents of formula (I) advantageously makes it possible to rapidly modify the surface of solid supports by a stable layer carrying easily sacrificial saccharide-type probe molecules of the support, given the presence of at least one ethylenic unsaturation on the spacer arm ^"X of the compounds of formula (I) according to the invention. When used in particular for the preparation of sugar chips, the compounds of formula (I) according to the invention have the following advantages:

- They play first of all as a spacer arm by allowing to move the saccharide chain away from the surface of the solid support that supports this chain. The unit X of the compounds of formula (I) according to the invention allows in particular the connection with a very wide range of saccharides, oligosaccharides or polysaccharides. In particular, the compounds of formula (I) can be used either as spacer arm of a first saccharide unit, for example an oligosaccharide unit, which will be attached to unit X and then it is possible to grow (combinatorial chemistry on solid support) or for the attachment of presynthesized saccharide probe molecules (attachment to the X unit, in the anomeric position of their reducing part).

- The spacer arm is cleavable: thanks to the presence of at least one ethylenic unsaturation on the unit X compounds of formula (I) that can be easily opened and targeted to isolate the sugar from the solid phase, and this under conditions that do not alter the integrity of the saccharide probe molecule. In particular, ozonolysis, Grubbs metathesis or dihydroxylation followed by oxidative cleavage of diol-osmylation (OsO ₄ , NaIO ₄ ) as well as other known mild chemical cleavage reactions can be used as the cleavage method. the skilled person.

Finally, because of their chemical functionalities and the choice of substituents on their saccharide backbone, the compounds of formula (I) according to the invention remain inert under many experimental conditions during the reactions carried out.

The present invention also relates to a method for preparing a solid support functionalized by probe molecules of saccharide nature, characterized in that it comprises at least one step of silanization of at least one surface of a -solid support -with a solution of at least one silanizing agent of formula (I) in an organic solvent. The organic solvent is preferably chosen from trichlorethylene, toluene, and lower alcohols such as ethanol or methanol, these solvents optionally being supplemented with a basic compound such as triethylamine or N ₅ N-diisopropylethylamine (DIEA). ). The contacting of the solid support with the solution of the silanizing agent of formula (I) is preferably carried out at a temperature of between 4 and 80 ° C., for about 1 to 48 hours.

The substrate is then rinsed with the reaction solvent or with chloroform and dried, preferably with nitrogen. This method has the advantage of being simple to implement and to combine the silanization step with the step of functionalization of the solid support, whereas the methods known from the prior art required at least three successive steps; that is to say a first step of silanization of the surface of the solid support with a molecule having a functional group allowing the attachment of a spacer arm in a second step and finally the attachment of a saccharide probe molecule in a third step, for example according to a glycosylation reaction. In this case, a step of inactivation ("capping" non-glycosylated sites) was then necessary, which is not the case according to the method of the present invention.

The solid supports that can be functionalized with the silanizing agents of formula (I) according to the invention are preferably chosen from glass, silica or other materials known to those skilled in the art as being capable of being used. silanized.

These solid supports have at least one flat surface or not, smooth or structured, and may be for example in the form of blade, flat or well plate, capillary or porous ball or not.

The present invention therefore also relates to solid supports characterized in that they comprise at least one functionalized surface with one or more silanizing agents of formula (I) as defined above.

Such carriers constitute sugar chips which are for example capable of being used for the identification, by screening, of saccharide molecules and in particular oligosaccharide sequences recognizing a particular protein of interest, for example using the method described in international application WO-A-03/008927.

In contrast, the sugar chips according to the present invention can also be used for the identification, by screening, of ligands, for example protein ligands recognizing a saccharide of interest.

Consequently, the subject of the present invention is finally a method for screening saccharide molecules, and in particular oligosaccharide sequences, or, respectively, protein ligands, characterized in that it comprises at least one contacting step of a solid support comprising at least one surface functionalized with at least one silanizing agent of formula (I) as defined previously with a solution containing one or more potential oligosaccharide molecules or, respectively, one or more potential protein ligands. In these particular applications, the functionalized solid supports in accordance with the present invention make it possible to optimize the screening methods and thus to dispose more efficiently and more rapidly of molecules for therapeutic or biotechnological purposes.

In addition to the foregoing, the invention also comprises other provisions which will emerge from the description which follows, which refers to an example of preparation of a compound of formula (I) according to the invention as well as an example of functionalization of a solid support with a compound of formula (I) according to the invention.

It should be understood, however, that these examples are given solely by way of illustration of the subject of the invention, of which they in no way constitute a limitation. EXAMPLE 1 Preparation of the Compound of Formula (1-1)

in which Ac represents acetyl.

1) First Step: Preparation of a spacer arm (3) and a glucose derivative (5)

The preparation of the spacer arm (3) is carried out according to the following reaction scheme A:

SmI ₂ , THF

SCHEME A

The spacer arm (3) is obtained in two successive steps: 2-vinyl-3-chloro-tetrahydrofuran (2) is accessible from 2,3-dichlorotetrahydrofuran (1) by treatment with a Grignard reagent according to the method described by L. Crombie, and RD

Wyvill, Journal of the Chemical Society, 1985, Perkin Trans 1, 1971 and references cited. The compound. (2) is then refluxed in tetrahydrofuran in the presence of 4-7 equivalents of Samarium diiodide (SmI 2) for 5 to 165 hours, according to the method described by L. Crombie, and LJ Rainbow, 1988, Tetrahedron Letters, 29 (49), 6517.

The compound (3) is obtained in a yield of 93%. The preparation of the glucose derivative (5) is carried out according to the following reaction scheme B:

SCHEME B

The glucose derivative (5) (thioglycoside) is obtained from D-Glucose in two successive stages which are conventional reactions of the chemistry of sugars:

a first step of peracetylation in an acetic anhydride / sodium acetate medium (Ac ₂ OZAcONa) at 120 ^° C. for 1 hour (the product obtained after precipitation in ice water is recrystallized in 95% ethanol) according to the process described by K. Takeo, Carbohydrate Research, 1980, 87, 147. The peracetylated by-product is obtained with a yield of 90%, the configuration of the anomeric position is predominantly β;

a second step of activating the anomeric position in anhydrous dichloromethane in the presence of thiophenol and a Lewis acid, in this case boron trifluoride etherate (BF ₃ -Et ₂ O), with at room temperature for one hour according to the method described by A. K Choudhury, N. Roy, Synthetic Communications, 1996, 26, 3937. This thioalkylation makes it possible to reach the thioglycoside (5), after recrystallization in methanol, and quantitatively . The thiophenyl group is in position β because of the presence of a group participating in position 2. 2) Second step: Preparation of a glycosylated spacer arm (6)

This glycosylated spacer arm is prepared according to the following Reaction Scheme C:

SCHEME C

The coupling of these two molecules is a glycosylation reaction of a protected glucose on an unsaturated chain.

The thioglycoside (5) (500 mg, 1.13 mmol, 1 eq.) And the unsaturated spacer arm (3) (127 mg, 1.13 mmol, 1 eq.) Are dissolved in anhydrous dichloromethane, in the presence of sieves. Molecular (630 mg). This mixture is stirred for 30 minutes at room temperature, then the reaction medium is brought to a temperature of -30 ^° C. N-iodosuccinimide (NIS) (510 mg, 2.26 mmol, 2 eq.) Is then added, followed by trifluoromethanesulfonic acid (30 μl, 0.34 mmol, 0.15 eq / NIS). The mixture is then slowly cooled to room temperature with stirring for 30 minutes. The reaction medium is then neutralized with a saturated aqueous solution of sodium hydrogencarbonate and then filtered on celite. The organic phase is extracted, washed successively with water, with an aqueous saturated solution of sodium thiosulfate and with an aqueous saturated solution of sodium chloride. The combined organic phases are dried over magnesium sulphate, filtered and concentrated under empty. The solid residue obtained is purified by chromatography on a column of silica gel. The product (6) is obtained with a yield of 70%. 2) Third step: Preparation of the compound of formula fl-1)

This step is carried out according to the following Scheme D:

SCHEMA D

During this step, 100 mg of the compound (5) obtained above in the preceding step (2.26 × 10 ^-4 mol, 1 eq.) Were reacted with 71 μl (3.62 × 10 ^-4 mol), 6 eq.) of triethoxysilane in the presence of 2.84 .mu.l (4.5.10 ^"4 mole) of Karstedt catalyst for 3 hours at a temperature of about 50 ⁰ C to yield a colorless liquid compound of formula (6), with a yield of 60%.

EXAMPLE 2 PREPARATION OF A FUNCTIONALIZED SOLID SUPPORT WITH A FORMULA SILANIZATION AGENT (I)

In this example, a planar SiO2 substrate was used. I) Substrate rehydration (Brown rehydration)

An SiO 2 substrate was soaked for 2 hours at room temperature in a mixture of deionized water (15 ml) and absolute ethanol (20 ml) containing 5 g of NaOH.

The substrate then _. It was washed with deionized water and then soaked for 1 hour in 0.2 N hydrochloric acid. was again washed with deionized water and then dried in an oven at a temperature of 80 ^° C. for 30 minutes

2) Silanization of the Substrate with the Compound of Formula (1-1)

The rehydrated substrate was soaked in 10 ml of a solution of trichlorethylene (TCE) containing 10 mM (21 mg) of compound of formula (I-1) as prepared in Example 1 above. After 1 night at ambient temperature, the substrate was silanized with the compound of formula (I-1) according to the invention.

The thus functionalized substrate was then washed with TCE, with ethanol and finally with chloroform. It was then dried in an oven for 30 minutes at a temperature of 50 ^° C.

For later use, the functionalized substrate was stored under an inert atmosphere (argon or nitrogen).

This substrate can then be used for the preparation of a sugar chip (growth of oligosaccharides on the silanized substrate thus prepared) or of any other molecule chip or biomolecule (in this case, the sweet silane which functions the substrate becomes a spacer for the attachment of any new molecule or biomolecule, natural or synthetic).

Claims

A silanization agent with a saccharide terminal function, characterized in that it corresponds to the following formula (I): A-X-B (I) in which:

the unit A represents a probe molecule of saccharide nature;

the unit X represents a spacer arm consisting of a carbon or heterocarbon chain comprising two ends, one of its two ends connecting, covalently, said spacer arm X to A and the other end connecting, covalently said spacer arm X has B, said chain having at least one ethylenic unsaturation between its two ends; it being understood that said chain can not contain several acetylenic unsaturations;

- B is a silanized group. 2. silanization agent according to claim 1, characterized in that the probe molecule of saccharide nature has a molecular weight of between 180 and 10 000 g / mol.

3. silanization agent according to claim 1 or 2, characterized in that the probe molecule of saccharide nature is chosen from monosaccharides, oligosaccharides, polysaccharides, glycoconjugates, glycoproteins, glycolipids and glycolipoproteins.

4. silanization agent according to claim 3, characterized in that the monosaccharides are chosen from glucosamine, azidoglucosamine, D-ribose, D-xylose, L-arabinose, D-glucose, D-galactose , D-mannose, 2-deoxyribose, L-fucose, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, N-acetylneuraminic acid, D-glucuronic acid, L-iduronic acid, D-sorbitol and D-mannitol.

Silanization agent according to claim 3, characterized in that the. -are- oligosaccharides selected -among sucrose, lactose, fragments ^" heparan sulphates, saccharide fragments of heparin, chondroitin or sulphate dermatans and Lewis antigens.

6. silanization agent according to claim 3, characterized in that the poly-oligosaccharides are chosen from the saccharide fractions of heparan sulphates, heparin or chondroitin and dermatan sulphates.

7. silanization agent according to claim 3, characterized in that the glycoconjugates are selected from heparan sulphates, heparin, chondroitin and dermatan sulphates.

8. silanization agent according to claim 3, characterized in that the glycoproteins are selected from immunoglobulin G and hyaluronic acid.

9. silanization agent according to claim 3, characterized in that the glycolipids are chosen from galactosyl-ceramides, gangliosides and cerebrosides.

10. silanization agent according to any one of the preceding claims, characterized in that one or more hydroxyl functions and / or leads saccharide entities of the probe molecule are protected by one or more protective groups selected from acetyl groups benzyl and aryl, 2,2,2-trichloroethyloxycarbonyl, benzyloxycarbonyl, trichloroacetamidate, tert-butyloxycarbonyl, and fluoranylmethoxycarbonyl. Silanization agent according to any one of the preceding claims, characterized in that one or more of the hydroxyl and / or amine functions of the saccharide entities of the probe molecule are substituted with one or more hydrophobic groups chosen from benzyl groups. acetate, benzylidene, isopropylidene and phthalimide. 12. silanization agent according to any one of the preceding claims, characterized in that the covalent bonds ensuring the attachment of each of the ends of the chain constituting the spacer arm X units A and B are derived from the reaction between a function initially chemically carried by the precursor of the X-spacer arm and a complementary chemical function carried on the one hand by the probe molecule A and on the other hand by the silanized group B. 13. silanization agent according to claim 12, characterized in that said covalent bonds result from the reaction between a hydroxyl radical and a group selected from halogen atoms and phosphite groups, trichloroacetamidate, thioalkyl, phosphate, pentenyl, sulfoxide and xanthate. 14. silanization agent according to any one of the preceding claims, characterized in that the spacer arm X represents a linear or branched alkyl chain C ₂ -C 4 _O or aryl C ₆ -C ₄₀ , said chain comprising at least an ethylenic unsaturation which may optionally be interrupted by one or more heteroatoms chosen from oxygen, nitrogen, sulfur and silicon, and / or one or more functions chosen from amide, oxime and tertiary amine functions and / or possibly substituted by one or more substituents chosen from linear or branched C ₂ -C ₂₀ alkyl or C ₆ -C ₂₀ aryl chains, said chains optionally possibly being also interrupted by one or more heteroatoms chosen from oxygen, nitrogen and , sulfur and silicon. 15. silanisation agent according to any one of the preceding claims, characterized in that the silanized group B is chosen from the groups -Si (Rj) ₃ , -SiR] (R ₂ ) ₂ and -SiRjR ₂ R ₃ in which the radicals R ₁ , R ₂ and R ₃ , independently of one another, represent an atom of halogen, alkoxy-C ₄ alkyl Ci-C _4, amino or an ester radical. 16. silanization agent according to claim 15, characterized in that the silanized group is selected from trimethoxysilyl groups, triethoxysilyl, trimethylsilyl and triethylsilyl.

17. silanization agent according to any one of the preceding claims, characterized in that it is chosen from compounds of formula (I) in which:

A is chosen from monosaccharides, oligosaccharides and polysaccharides,

- X represents a carbon chain having 2 to 40 carbon atoms, comprising -at -moins- a éthyléniqύë unsaturated ^"said chain being linear or branched and optionally interrupted by one or more rings and / or one or more functions such as amide, oximes and tertiary amino functions;

B represents a trimethoxysilyl or triethoxysilyl group. 18. silanization agent according to any one of the preceding claims, characterized in that it is chosen from the following compounds of formulas (I-1) and (1-2):

in which Ac represents the acetyl group. 19. Use of at least one silanizing agent of formula (I) as defined in any one of claims 1 to 18 for the functionalization of solid supports.

20. Use of at least one silanizing agent of formula (I) as defined in any one of claims 1 to 18, for the manufacture of sugar chips. 21. A process for preparing a solid support functionalized by probe molecules of saccharide nature, characterized in that it comprises at least one step of silanizing at least one surface of a solid support with a solution of at least one silanising agent of formula (I) as defined in any one of claims 1 to 18 in an organic solvent. _ .. 20 - -22-. -Strong support characterized by the fact that ^" it comprises at least one surface functionalized with one or more silanisation agents of formula (I) as defined in any one of claims 1 to 18.

23. Use of a solid support as defined in claim 22 for the identification, by screening, of oligosaccharide sequences recognizing a protein of interest or ligands recognizing a saccharide of interest.

24. A method for screening saccharide molecules or, respectively, protein ligands, characterized in that it comprises at least one step of contacting a solid support as defined in claim 22 with a solution containing one or several potential oligosaccharide molecules or, respectively, one or more potential protein ligands.