FR2664813A1 - Compositions based on hydrophobic substances and which can be solubilised in an aqueous solvent, process for obtaining them and their uses, in particular in the pharmaceutical field - Google Patents

Abstract

The invention relates to any composition which comprises: - on the one hand a water-insoluble pharmacologically active substance, this substance being substituted with at least one lipophilic group, and - on the other hand a glycoside derivative, at least one of the hydroxyl functions of which is substituted with one or more phosphate or sulphate group(s), this compound also being substituted with a lipophilic group, this composition being capable of being solubilised or dispersed homogeneously in an aqueous solvent. The invention also relates to the use of such compositions, in particular as pharmaceutical compositions in solution in a physiologically acceptable aqueous solvent.

Description

COMPOSITIONS BASED ON HYDROPHOBIC SUBSTANCES, SOLUBILIZABLE IN AQUEOUS SOLVENT, PROCESS THEREOF
OF OBTAINING, AND THEIR USES IN PARTICULAR IN THE
PHARMACEUTICAL FIELD.

 The invention relates to compositions comprising hydrophobic substances, however, solubilizable in an aqueous solvent.

 The invention also relates to the process for obtaining such compositions, and their uses in particular as pharmaceutical compositions.

 Many products used in the pharmaceutical field, or likely to be, can not be solubilized in water.

 This insolubility in water is particularly disadvantageous for products intended to be administered in an aqueous physiological solution of the type usually used for injectable preparations.

 Among these pharmacologically active products which are insoluble in water, mention may be made in particular of lipophilic derivatives of muramyl peptides, such as those described in European Patent No. 0 165 123 of May 10, 1985, or in the patent 4512 of 20/03/1979.

 In fact, these products are immunomodulators capable of increasing the specific immune response (vaccine adjuvant), or of enhancing non-specific resistance (anti-infective agent).

 Being of a very hydrophobic nature, they give aqueous suspensions difficult to disperse in a completely satisfactory manner (especially with regard to the level of dispersion compatible with the possibility of injecting them into a patient), by using the various means of dissolution or aqueous dispersion based on known techniques, including mechanical treatments, or the use of organic solvents or surfactants.

 The present invention is specifically intended to provide means for the solubilization or homogeneous dispersion of hydrophobic products, such as lipophilic muramyl-peptides, in an aqueous solvent.

 The invention generally relates to any composition characterized in that it comprises - on the one hand a pharmacologically active substance, insoluble (in whole or in part) in water, this substance being substituted by at least one group lipophilic, and - on the other hand, a saccharide derivative at least one of the hydroxyl functions is substituted by one or more phosphate or sulfate group (s), this derivative also being substituted by at least one lipophilic group, this composition being capable of being solubilized or dispersed homogeneously in an aqueous solvent.

 The pharmacologically active substance is non-covalently associated with the osidic derivative via their respective lipophilic group (s).

 Advantageously, the two lipophilic groups ensuring this non-covalent association are such that one of them is composed of a linear hydrocarbon structure, while the other is a branched hydrocarbon structure.

 The branched hydrocarbon structure is preferably chosen from those consisting of a branched hydrocarbon chain, whose branches originate on two adjacent carbon atoms, or at most separated from a carbon atom.

The linear hydrocarbonaceous structure must have, in order to associate with the branched hydrocarbon chain, a sufficient length allowing the introduction of non-covalent bonds, in particular of the Van der type.
Walls, between the linear hydrocarbon structure and at least one branch of the branched hydrocarbon structure.

 Thus, the combination of the pharmacologically active substance and the osidic derivative is effected via their respective lipophilic groups, provided that non-covalent bonds are established which are strong enough to allow the linear hydrocarbon structure to be maintained in a position called "sandwich" between the two branches of the branched hydrocarbon structure.

 Preferably, the linear hydrocarbon structure has a number of carbon atoms at most equal to the number of carbon atoms of the branches of the branched hydrocarbon structure.

 The combination of the saccharide derivative as described above, of an amphiphilic and water-soluble nature, with the hydrophobic and water-insoluble pharmaco logically active substance, allows the homogeneous dispersion of the latter in an aqueous solvent.

The invention more particularly relates to any composition capable of being solubilized or dispersed homogeneously in an aqueous solvent, characterized in that it comprises (1) a pharmacologically active substance, insoluble in water, this substance being substituted
or by at least one linear hydrocarbonaceous structure of 4 to 20 carbon atoms, optionally comprising an ether, ester, thioether, thioester or amide linkage, this structure being designated hereinafter as a lipophilic group of type A,
or at least one branched hydrocarbon structure of 10 to 50 carbon atoms, optionally comprising one or more ether, ester, thioether, thioester or amide linkage, this structure being designated hereinafter by lipophilic group of type B, (2) a compound consisting of a saccharide derivative non-covalently associated with the substance (1), this osidic derivative being itself substituted
or by at least one lipophilic group of type A when said pharmacologically active substance is substituted with a lipophilic group of type B,
or by at least one lipophilic group of type B when said pharmacologically active substance is substituted by a lipophilic group of type A, at least one of the hydroxyl functions of the above-mentioned saccharide derivative being substituted with one or more groups ( ) phosphate or sulphate.

 In general, any osidic derivative as defined above is usable in the compositions of the invention, since this derivative does not cause a modification of the pharmacological properties of the aforementioned active substance, such that the latter becomes inactive or toxic.

dans laquelle - R représente un groupement lipophile du type A ou du type B, - R2 représente H, ou OH, ou un groupement acetamido, sulfate, sulfamido, phosphate, phosphamido, - R3 et R4, identiques ou différents, représentent soit H, soit OH, ou OCH3, ou encore un groupement sulfate ou phosphate, - R1 représente soit H, soit OH, ou OCH3, ou encore un résidu ose ou osamine dont le carbone anomérique en position 1 (C1) est engagé dans une liaison glycosidique, les positions C2, C3 et C4 étant respectivement substituées par des groupes R'2, R'3, et R'4 ayant respectivement les significations données ci-dessus pour R2, R3, et R4, et la position C6 par un groupe R' ayant la signification donnée ci-dessus pour
R, l'un au moins des groupes R2, R'2, R3, R'3, R4 ou R'4 représentant un résidu sulfate ou phosphate.Particularly preferred compositions of the invention are those comprising osidic derivatives corresponding to the following formula (I)

in which - R represents a lipophilic group of type A or of type B, - R2 represents H, or OH, or a group of acetamido, sulphate, sulphonamido, phosphate, phosphamido, - R3 and R4, which are identical or different, represent either H, either OH, or OCH3, or else a sulphate or phosphate group, - R1 represents either H, or OH, or OCH3, or a dose or osamine residue whose anomeric carbon in position 1 (C1) is engaged in a glycosidic bond, the C2, C3 and C4 positions being respectively substituted by R'2, R'3, and R'4 groups respectively having the meanings given above for R2, R3, and R4, and the C6 position by a group R '; having the meaning given above for
R, at least one of R2, R'2, R3, R'3, R4 or R'4 representing a sulphate or phosphate residue.

By way of example of the saccharide derivatives corresponding to the above-mentioned formula (I), mention may be made of the following compounds: α- or β-methyl-glycoside N-acetyl-4-sulfate-6-OR-2-desoxy-glucosamine N-acetyl 4-sulphate-6-OR-2-desoxy-glucosamine, α- or 8-methyl-glycoside 4-sulphate-6-OR-glucopyranose, 4-sulphate-6-OR-glucopyranose, -6 , 6'-di-OR-4,4a-di-sulfate-a, a'-D-trehalose,
R having the meaning indicated above.

Preferred compositions of the invention are those comprising the following osidic derivatives of formula (I) -a or P-methyl-glycoside N-acetyl-4-sulphate-6-octanoyl-2-desoxy-glucosamine, -a or ss-methyl glycoside-N-acetyl-4-sulphate-6- (2,3-dipalmitoyl) glyceroyl-2-desoxy-glucosamine, -N-acetyl-4-sulphate-6-octanoyl-2-desoxy-glucosamine, -N acetyl-4-sulphate-6- (2,3-dipalmitoyl) -glyceroyl-2-desoxy-D-glucosamine, α- or β-methyl-glycoside 4-sulphate-6-octanoyl-glucopyranoside, -α or β- methyl-glycoside 4-Sulfate-6- (2,3-dipalmitoyl) -glyceroyl-D-glucopyranoside, 4-Sulfate-6-octanoyl-D-glucopyranose, 4-Sulfate-6 (2,3-di-palmitoyl) ) -glyceroylglucopyranose - 4,4'-6,6-di-octanoyl-α, α'-D-trehalose, -4,4'di-6-sulphate 6 '(2,3-dipalmitoyl) - glyceroyl-a-a'
D-Trehalose.

In general, any pharmacologically active principle of an insoluble nature in an aqueous solvent, and on which is capable of being grafted at least one lipophilic group of type A or
B above-mentioned, may be used as a pharmacologically active substance in the compositions of the invention.

dans laquelle R, représente H ou CH3 - R6 représente un groupe -NH2, -OH ou un groupe -OW1,
W1 étant un groupe hydrocarboné comportant de 1 à 10 atomes de carbone - X représente un résidu aminoacyle du groupe comprenant alanyle, valyle, isoleucyle, norleucyle, leucyle, thréonyle, prolyle, glutaminyle, asparaginyle, méthionyle, tryptophanyle, phénylalanyle, tyrosyle, glycyle - Y représente soit OH, soit NH2, ou un groupe -OW2, W2 étant un groupe hydrocarboné de 1 à 4 atomes de carbone, ou encore un groupement lipophile du type A ou du type B, - Z représente soit H, soit un groupement lipophile du type A ou du type B, étant cependant entendu que l'un au moins des deux groupes Y et Z est toujours constitué par un groupement lipophile du type A ou du type B - n1 et n2, identiques ou différents, représentent zéro ou 1, - A1 et B1 sont soit des groupes respectivement identiques ou différents comprenant de 1 à 3 résidus aminoacyles, eux-mêmes identiques ou différents entre eux, ou encore un groupe -NH-(CH2)p-CO- avec des valeurs de p comprises entre 2 et 10.Among the pharmacologically active substances used in the compositions of the invention, mention may be made of the lipophilic derivatives of muramyl-peptide mentioned above, and corresponding to the following formula (II)

wherein R, is H or CH3 - R6 is -NH2, -OH or -OW1,
W 1 being a hydrocarbon group having from 1 to 10 carbon atoms - X represents an aminoacyl residue of the group comprising alanyl, valyl, isoleucyl, norleucyl, leucyl, threonyl, prolyl, glutaminyl, asparaginyl, methionyl, tryptophanyl, phenylalanyl, tyrosyl, glycyl - Y represents either OH or NH2, or a group -OW2, W2 being a hydrocarbon group of 1 to 4 carbon atoms, or a lipophilic group of type A or type B, - Z represents either H or a lipophilic group of type A or type B, it being understood, however, that at least one of the two groups Y and Z is always constituted by a lipophilic group of type A or of type B - n1 and n2, which are identical or different, represent zero or 1 And A1 and B1 are either respectively identical or different groups comprising from 1 to 3 aminoacyl residues, themselves identical or different from each other, or alternatively a group -NH- (CH 2) p -CO- with values of p included between 2 and 10.

A particularly preferred composition of the invention comprises compounds of the above-mentioned formula (II), in which - n1 and n2 represent zero, - R5 represents CH3, - R6 represents -OH, -NH2 or -On (CH2) H with x1 being x1 between 1 and 6, with a preference for x1 = 4, -X is L or D-alanyl, L-threonyl, L-valyl, - when Y is -O-CH2-CHO (R7) -CH2O ( R8)
Z represents H, and when Z represents O
|
-C-CHO (R7) -CH2O (R8), Y is -OH, -NH2 or a group -OW2I W2 having the meaning indicated above, and R7 and R8 are palmitoyl groups.

 Another preferred composition of the invention comprises compounds of formula (II) above-mentioned, wherein - n1 and n2 represent zero, - R5 represents CH3 - R6 represents -OH, or -NH2 or On (CH2) xrH with x1 between 1 and 6, with a preference for x1 = 4, -X is L or D-alanyl, L-threonyl, L-valyl, - when Y is -O- (CH2) x2H with x2 = 8, Z is H and when Z is -CO (CH2) x2H, where x2 = 8, Y is -OH, -NH2, or -OW2, W2 having the meaning indicated above.

 The subject of the invention is also the osidic derivatives of formula (I) and also their process for obtaining according to methods which are conventionally used in synthetic chemistry of oses and oligosaccharides.

 As an illustration, this methodology proceeds in successive steps from the saccharide or oligosaccharide to be chemically modified, to obtain the desired derivative, specifically substituted on the desired position. For this purpose, the functions in positions C1, C2, C3, C4 and C6 of a saccharide and those corresponding to an oligosaccharide are protected temporarily by protecting groups (such as benzyl, benzylidene, acetyl, etc.) introduced specifically. and sequentially on the positions that will not ultimately be the subject of the proposed chemical modification, taking advantage of the differences in reactivity of said functions.

 Then the introduction of the lipophilic groups of the type A or B mentioned above, is advantageously carried out in the following manner the unprotected primary hydroxyl groups (or la), can react either in the form substituted with a tosyl group with an acid salt in the presence of crown ether, either in free form with a fatty acid chloride, or with the carboxylic function of a fatty acid in the presence of a coupling reagent.

 After introduction of the (or) chemical modifications envisaged on the (or) free positions, the derivative obtained is released (deprotection step) of all its protective groups to result in the desired osidic or oligosaccharide derivative.

Where appropriate, the introduction of (or) phosphate groups is carried out prior to the aforementioned deprotection step, by the so-called phosphite triester method described in Perich, JW et al,
Tetrahedron Lett., (1988), 29, 2369.

 The introduction of (or of) sulphate groups is carried out after the above-mentioned deprotection step, in particular by using the sulfuric trimethylamineanhydride complex according to the method especially described in Ichikawa Y. et al., Carbohydrate Research, 172, (1988). , 37-64.

 The invention also relates to pharmaceutical compositions comprising a composition of the type of those described above, or a mixture of these compositions, dispersed in a physiologically acceptable aqueous solvent.

 Advantageously, the compositions of the invention may be lyophilized from their solution or aqueous dispersion, and are in the form of a powder.

 These dry powder compositions are readily solubilized or dispersed again in a suitable volume of aqueous solvent.

 The invention also relates to a method for modifying the solubility properties of a pharmacologically active substance of the type (1) described above, this process consisting of the combination of the substance (1) with a type 2 (2) ) as described above, under conditions such that when this combination of (1) and (2) is suspended in water, a solution (or dispersion) is obtained which can be used especially as an injectable preparation, when the aforementioned water is sterile.

 The solubilization of the pharmacologically active substance (1) is such that a fluid characterized by a certain opalescence (measurable by nephelemetry) is obtained.

 This fluid behaves like a solution and can be injected into the body just like an injectable solution.

 The subject of the invention is also a process for obtaining the compositions described above comprising the solubilization of the osidic derivative (s) in a given amount of an aqueous solvent, the stirring addition to the solution obtained previously. , a determined amount of one (or more) pharmacologically active substance, - if appropriate, lyophilization of the previous solution.

 In a variant, the process for obtaining the compositions of the invention is carried out by adding, with agitation, the osidic derivative (s) to an aqueous suspension comprising a determined quantity of one (or more) pharmacologically active substance.

 The invention is further illustrated in the detailed description which follows, without the latter being limited in nature.

1) Description of the synthesis of the sodium salt of 6,6'-octanoyl-4,4'-disulfonyl-α, α'-D-trehalose (or compound P1) .6,6-octanoyl-2,3- 2,3 ', tetra-O-benzyl-α, α'-D-trehalose
Dissolution in dry THF (50 ml), under argon, protected from the light of 3 mmol of 2,3-2 ', 3' tetra-O-benzyl-α, α'-D-trehalose (AF Hadfield ,
L. Hough, and AC Richardson - Carbohyd. Res. 63, (1978), 51) of 7.5 mmol of octanoic acid, and 7.87 mmol of triphenylphosphine. To the cooled solution at 0 ° C, addition in 15 minutes of a solution of diisopropylazodicarboxylate (8.02 mmol) in the
Dry THF (25 ml). Purification on silica 60 H under pressure in toluene / CH 2 Cl 2 / AcOET 30/10 / 5. Oil: m = 2.58 g (90%).

 Sodium salt of 6,6'-dioctanoyl-2,3-2 ', 3'-tetra-O-benzyl-4,4'-di-sulfonyl-α, α'-D-trehalose.

 To 1.5 mmol of the above product dissolved in dry DMF (15 ml), addition of 15 mmol of trimethylamine / sulfur trioxide complex (2.1 g) and incubation at 50 ° C under anhydrous conditions for 2 days. CH 3 OH (10 ml), then extension with chloroform (10 ml) and percolation on Sephadex LH 2 O in CHCl 3 -CH 3 OH: 1 / 1. After concentration, purification on 60H silica column under pressure, in CHCl 3 -CH 3 OH mixture -H 2 O 15/5/05 (1.7 g: oil) Dissolution in the DMF-H 2 O: I / I mixture (80 ml) and percolation on a Dowex 50 WX4 H + resin column (85 ml) by elution with the same solvent mixture: neutralization with 0.1N NaOH: lyophilization m = 1.51 g (87%).

6,61-Dioctanoyl-4,41-di-O-sulfonyl-α, α-D-trehalose sodium salt (C28H48Na2 19S2: 798.79)
1.5 g of the preceding derivative, solubilized in 150 ml of a water / alcohol mixture (6.5 / 10), is hydrogenated in the presence of 750 mg of Pd / C at 10%. After filtration, concentration, lyophilization in aqueous solution.

Purification on Silica 60-G in CHC13-CH30H-CH3COOH
H20: 40/20/8/2. Concentration, coevaporation with toluene, lyophilization in aqueous solution.

Percolation on Sephadex LH in methanol.

Concentration desiccation, lyophilization in solution in water m = 525 mg (51%).

Sulphate content: 2.7 Eq / g (conductimetry).

octanoic acid content: 1.97 Eq / mole (CPV).

2> Description of the use of the aqueous dispersion of MDP-GDP, that is to say (N-acetyl-muramyl-L-alanyl-D-isoglutamine) -P, -dipalmitoyl-sn-Gly - cerol, with the compound P1
10 mg of P1 are dissolved in 10 ml of distilled water. To the solution are added 10 mg of CDM
GDP, under heavy agitation. The aqueous dispersion obtained is opalescent in appearance and does not sediment or very little, (further stirring makes it possible to obtain the homogeneous dispersion again). After lyophilization, it is possible, by addition of water or physiological solute, to reconstitute a dispersion of identical appearance, the concentration of derivatives depending on the volume of the addition.

 The method allows an easy galenic formulation of the muramyl peptides corresponding to the general formula (II). This results in preparations which retain the pharmacological activities of these products, particularly as a regulator of the immune response, as claimed in European Patent No. 0 165 123 3) Influence of the use of P1 on the adjuvant activity of lipophilic muramyl-peptides studied in guinea pigs.

a) Preparation of the emulsion
1 mg of P1 is dissolved in 0.5 ml of saline, this solution is added dropwise to 1 mg of powdered MDP-GDP.

The dispersion obtained is mixed with 0.5 ml of an isotonic solution of ovalbumin at 20 mg / ml. It is added 1 ml of mineral oil (Adjuvant
Incomplet de Freund, AIF) and proceed to the emulsion.

This preparation therefore contains, per ml, 0.5 mg of adjuvant, 0.5 mg of P1 and 5 mg of ovalbumin.

b) Methodology
The control batches receive a) the water / mineral oil emulsion containing the antigen alone, b) the same emulsion containing the adjuvant molecules alone (MDP-GDP). The experimental batches receive the emulsions containing the osidic drift P1. Injections are done subcutaneously in the rear plantar pads of male guinea pigs
Hartley weighing 350 g. After 3 weeks the animals receive intradermally 0.1 ml of saline alone or containing 0.1 ml of ovalbumin.

After 48 hours, the induration diameter is measured which is the sign of the establishment of a state of delayed hypersensitivity. After 24 hours, the animals are sacrificed by cardiac puncture and the sera collected. The titration of the circulating antibodies is carried out by the Elisa method according to the conventional methods, in particular those described in
F.AUDIBERT, L. CHEDID, P. LEFRANCIER and J. CHOAY,
Immunol. (1976), (21), 143; and M. JOLIVET, F.

AUDIBERT, H. GRAS-MASS, A. TARTAR, D. SCHLESSINGER,
R. WITZ and L. CHEDID, Infect. Immunol. (1987), (55), 1498.

c) Results
The results obtained are recorded in the following table
IMMUNIZATION * HYPERSENSITIVITY TITLE ANTI
DELAYED - BODY ELISA
diameter of indu
ration mm
AIF 0.0.0.0 22,000, 37,000,
65,000, 95,000
AIF + P1 0.0.0.0 7.000, 8.000, 100 pg 21,000.

AIF + MDP-GDP 100g 8,14,15,17 241,000,295,000
325.000,641.000.

AIF + MDP-GDP 5,16,20,25 157,000,275,000, 100pg + P1 275,000,318,0000.

100 Ag *: All animals received 1 mg ovalbumin.

The figures represent the maximum dilution of the serum making it possible to obtain an optical density equal to that obtained using a normal serum diluted 50 times.

 These results show that MDP-GDP is a strong adjunct to the cell-mediated response and the humoral response and that the addition of P1 did not modify their activity.

4) Influence of the P1 osidic derivative on the immunostimulatory properties of muramyl peptides.

a - Anti-infectious activity
Swiss mice received P1, MDP-GDP, or
MDP-GDP + P1 by venous route 24 hours before infection with 1 x 104 Klebsiella pneumoniae. The P1 solution is added to the MDP-GDP to obtain a preparation containing an equivalent amount of 2 products (weight / weight).

The results in Table I show that P1 alone has no protective action against infection. The comparison of the activity of the different doses of
MDP-GDP, alone or with P1, does not show significant differences.

It can be concluded from these experiments that the compound P1 does not inhibit the anti-infectious activity of the
MDP-GDP.

b - Action of NDP-GDP on TNF production
flowing
TNF tumor necrosis factor is a protein synthesized by cells of the monocyte / macrophage lineage when stimulated. The
LPS (bacterial lipopolysaccharide) is a very potent inducer of TNF production. Muramylpeptides stimulate TNF production in vitro but are significantly less effective than LPS. In vivo, serum TNF levels produced after the administration of muramyl-peptides are below or at the limit of the detection possibilities.

 MDP-GDP, and the other derivatives, which have an anti-infectious action, have the property of increasing the response to the LPS injected afterwards, causing a considerable increase in the level of serum TNF. Table II shows the results of an experiment of this type. It can be seen that the pretreatment is more effective 6 hours before the LPS, in particular in the case of the MDP-GPD + P1 combination, but that 16 hours before, the injection of MDP-GDP still has a potentiating effect on the reply.

c - Action of MDP-GDP in vitro on monocytes
human blood.

 Blood monocytes are isolated and incubated for 24 hours with different doses of product.

The supernatants are removed to assay for TNF and IL-6, cytokines that are produced by activated monocytes.

 Table III shows that the production of TNF is significant only with 10 μg / ml of MDP-GDP and that the product solubilized with compound P1 has a slightly higher activity. The IL-6 assay gives clearer results and also shows comparable monocyte stimulation between MDP-GDP and solubilized MDP-GDP in association with P1.

d - Action on human polynuclear
The muramyl-peptides have a direct action on the polynuclear which is demonstrated in particular by the phagocytosis and the destruction of Candida albicans.

 The estimation of this action is made by the ability to incorporate glucose treated by the viable mushroom cells.

In the two experiments reported on the
Table IV, unstimulated control cells already have a relatively high basal activity.

However, the addition of muramyl-peptide, with or without
P1, produces an increase in cell inhibitory activity on the growth of the fungus.

In conclusion, whether in experiments made in vivo in mice or in vitro with human cells, the solubilization of MDP-GDP by compound P1 never decreases its activity. The compound
P1 alone is still inactive.

TABLE I
Protective action of MDP-GDP solubilized by compound P1 in the mouse infected with Klebsiella pneumoniae
IV Treatment, Survival at D + 15% Protection *
at J-1
Witnesses 1/16
P1 100 yg 0/8
MDP-GDP 3 pg 3/8 37.5 ns
10 3/8 37.5 ns
30 12/16 68.7 P <0.01
100 4/8 37.5 ns
CDM-PDM / P1
3 / 3g 3/8 37.5 ns
10/10 3/8 37,5 ns
30/30 8/16 43.7 P <0.02
100/100 7/8 75 P <0.02 *: Statistical calculations are made taking into account
account for each batch of their witnesses
respectively.

TABLE II
Influence of P1 solubilized MDP-GDP pretreatment on the LPS-induced TNF response
Treatment with 100g LPS iv TNF activity in iv / mouse before LPS 25 g serum
U / ml pg / ml h-6 P1 + 189 3.4 h-6 MDP-GDP / P1 - <10 <0.18 h-6 MDP-GDP + 11.475 206.55 h-6 MDP-GDP / P1 + 19,038 342.68 h-16 MDP-GDP + 1.899 34.18 h-16 MDP-GDP / P1 4.296 77.33 *: 2 hours after challenge injection with LPS
Activity expressed in U / ml and murine recombinant TNF equivalent in the cytotoxicity assay of the L292 cell line.

TABLE III
Action of MDP-GDP in vitro on human monocytes
Stimulant g / ml Activity in the supernatant
TNF U / ml IL-6 U / ml
P, O <10
MDP-GDP 0.1 0 <10
0.1 0 <10
1 0.3 <10
10 6.9 94.9
MDP-GDP / P1 0.01 0 <10
0.1 0 <10
1 5.9 83
10 10.5 594
Cells at 2 x 10 6 / ml, incubated for 24 hours with the products.

TABLE IV
MDP-GDP action on human polynuclear
Stimulant Zg / ml Inhibition of growth of
Candida (%)
1st exp. 2nd exp.

Witnesses 35.3 41.5 P1 10 34.6 38.7
MDP-GDP 0.1 44.1 47.2
75.1 78.1
10 86.2 86.1
MDP-GDP / P1 0.1 32.2 77.2
1 41.4 93.6
10 65.9 95.1
The polynuclear cells (1 x 10 6 / ml) are incubated with the stimulants for 30 minutes and distributed in the wells of the microplates with a suspension of C.

albicans at 1 x 104 / ml. After 18 hours, the culture medium is removed and the incorporation of 3H-D-glucose by the Candida cells is measured after a 3 hour incubation.

5) Influence of the P1 glycidyl derivative on the pyrogenic effect of MDP-GDP.

 The pyrogenicity tests were carried out on New Zealand rabbits of 3 to 4 kilograms maintained in a box of contention. The animals received a dose of product in a volume of 0.5 ml / kg by the venous route at zero time and their temperature recorded for 5 hours continuously.

1. Evaluation of the Polypropenicity of P1
Two batches of three rabbits were constituted group I: 10 g / kg of P1; group II: 100 μg / kg P1.

The recorded temperature differences were for group I: 0.15, 0.25; 0.25 (C), an average of: 0.22 "C + 0.06, for group II: 0.40, 0.15, 0.30 (" C), an average of: 0.28 C + 0.12.

 This experiment indicates that P1 is totally devoid of pyrogenic effect in rabbits and in the drastic conditions of a recording carried out for 5 hours.

2. Evaluation of the pyrogenicity of MDP-GDP in
presence of P1
The minimum pyrogenic dose of MDP-GDP previously determined is equal to: 2 Ag / kg.

 A dose of 10 Ag / kg was chosen which gives approximately a temperature rise of 1 C.

 Two batches of three rabbits were constituted group I: 10 Sg / kg of MDP-GDP; group II: 10 μg / kg of MDP-GDP + 10 μg / kg of P1.

 The recorded temperature differences are for group I: 0.70, 0.50, 0.85 (C), an average of: 0.68 C + 0.18; for group II: 1.15, 0.90, 0.65 (C), an average of: 0.900C + 0.25.

 A Student statistical test (unpaired twotailed test, p = 0.2866) indicates that this difference is not significant and therefore that P1 does not synergize the pyrogenic effect of another product such as MDP-GDP.

Claims (12)

 1. A composition capable of being solubilized or dispersed homogeneously in an aqueous solvent, characterized in that it comprises (1) a pharmacologically active substance, insoluble in water, this substance being substituted  or by at least one linear hydrocarbonaceous structure of 4 to 20 carbon atoms, optionally comprising an ether, ester, thioether, thioester or amide linkage, this structure being designated hereinafter as a lipophilic group of type A,  or at least one branched hydrocarbon structure of 10 to 50 carbon atoms, optionally comprising one or more ether, ester, thioether, thioester or amide linkage, this structure being designated hereinafter by lipophilic group of type B, (2) a compound consisting of a saccharide derivative non-covalently associated with the substance (1), this osidic derivative being itself substituted:  or by at least one lipophilic group of type A when said pharmacologically active substance is substituted with a lipophilic group of type B,  or by at least one lipophilic group of type B when said pharmacologically active substance is substituted by a lipophilic group of type A, at least one of the hydroxyl functions of the above-mentioned saccharide derivative being substituted with one or more groups ( ) phosphate or sulphate.  2. Composition according to claim 1, characterized in that the osidic derivative is represented by the following formula (I)

 in which - R represents a lipophilic group of type A or of type B, - R2 represents H, or OH, or a group of acetamido, sulphate, sulphonamido, phosphate, phosphamido, - R3 and R4, which are identical or different, represent either H, either OH, or OCH3, or else a sulphate or phosphate group, - R1 represents either H, or OH, or OCH3, or a dose or osamine residue whose anomeric carbon in position 1 (C1) is engaged in a glycosidic bond, the C2, C3 and C4 positions being respectively substituted by R'2 groups, R'3 r and R'4 respectively having the meanings given above for R2, R3, and R4, and the C6 position by a group R '; having the meaning given above for R, at least one of R2, R '2 R3, R'3, R4 or R'4 representing a sulphate or phosphate residue.  3. Composition according to Claim 1 or Claim 2, characterized in that the saccharide derivative is chosen from the following compounds - α or β-methyl-glycoside N-acetyl-4-sulphate-6-OR-2-deoxy-glucosamine, N-acetyl-4-sulphate-6-OR-2-desoxy-glucosamine, α- or β-methyl-glycoside 4-sulphate-6-OR-glucopyranose, -4-sulphate-6-OR-glucopyranose, -6, 6'-di-OR-4,4'-di-Sulfate- &alpha;, &alpha; '- D-trehalose, R having the meaning indicated in claim 2.  4. Composition according to one of claims 1 to 3 characterized in that the osidic derivative is selected from the following compounds: - a or P-methyl-glycoside N-acetyl-4-sulphate-6octanoyl-2-desoxy-glucosamine, - &alpha; or ss-methyl-glycoside-N-acetyl-4-sulphate-6- (2,3-dipalmitoyl) glyceroyl-2-desoxy-glucosamine, - N-acetyl-4-sulphate-6-octanoyl-2-desoxy-glucosamine N-acetyl-4-sulphate-6- (2,3-dipalmitoyl) -glyceroyl-2-desoxy-D-glucosamine, α-β-methyl-glycoside 4-sulphate-6-octanoyl-glucopyranoside, - or p-methyl-glycoside 4-sulphate-6- (2,3-dipalmitoyl) -glyceroyl-D-glucopyranoside, 4-sulphate-6-octanoyl-D-glucopyranose, 4-sulphate-6 di-palmitoyl) -glyceroylglucopyranose-4,4'-6,6-di-octanoyl-α, α'-D-trehalose, 4,4'di-6,6-diisulfate (2,3-di-octanoyl) dipalmitoyl) does -glyceroyl D-Trehalose.  5. Composition according to claim 1, characterized in that the pharmacologically active substance is a lipophilic derivative of muramyl-peptide that can be used as a vaccine adjuvant or an anti-infective agent.  6. Composition according to Claim 5, characterized in that the pharmacologically active substance is chosen from compounds corresponding to the following formula (II)

 in which - N represents H or CH3; - R6 represents a group - NH2, -OH or a group -OW1, W1 being a hydrocarbon group having from 1 to 10 carbon atoms; X represents an aminoacyl residue of the group comprising alanyl, valyl, isoleucyl, norleucyl, leucyl, threonyl, prolyl, glutaminyl, asparaginyl, methionyl, tryptophanyl, phenylalanyl, tyrosyl, glycyl-Y represents either OH or NH2, or a -0W2 group; W 2 being a hydrocarbon group of 1 to 4 carbon atoms, or a lipophilic group of type A or type B, Z represents either H or a lipophilic group of type A or type B, it being understood, however, that at least one of the two groups Y and Z is always constituted by a lipophilic group of type A or of type B; - n1 and n2, identical or different, represent zero or 1, - A1 and B1 are either respectively identical or different groups comprising from 1 to 3 aminoacyl residues, themselves identical or different from each other, or a group -NH- (CH2) p-CO- with p values between 2 and 10.  7. Composition according to Claim 6, characterized in that the pharmacologically active substance is chosen from compounds of formula (II) in which - n1 and n2 represent zero, - R5 represents CH3, - R6 represents -OH, -NH2 or -On (CH2) XH with x1 between 1 and 6, with a preference for x1 = 4, - X is L or D-alanyl, L-threonyl, L-valyl, - when Y is -O-CH2-CHO (R7) -CH2O (R8),  Z represents H, and when Z represents

 Y Y represents -OH, -NH 2 or a group -OW 2 W 2 having the meaning indicated in claim 6, and R 7 and R 8 are palmitoyl groups.  8. Composition according to claim 6, characterized in that the pharmacologically active substance is chosen from compounds of formula (II) in which - n1 and n2 represent zero, - R5 represents CH3, - R6 represents -OH, or -NH2 or -On (CH2) 1H with x ranging from 1 to 6, with a preference for x1 = 4, - X is L or D-alanyl, L-threonyl, L-valyl, - when Y is - - (CH2) x2H with x2 = 8, Z 2 represents H, and when Z represents -CO (CH2) x2H, with x2 = 8, Y represents -OH, -NH2, or a group -OW2, W2 having the meaning indicated in claim 6.  9. Pharmaceutical composition characterized in that it comprises a composition according to any one of claims 1 to 8 dispersed in a physiologically acceptable aqueous solvent.  Derivative having the following general formula (I):

 in which - R represents a group of type A or type B, - R2 represents H, or OH, or a group of acetamido, sulphate, sulphonamido, phosphate, phosphamido, - R3 and R4, which are identical or different, represent either H or OH, or OCH3, or a sulphate or phosphate group, - R1 represents either H, or OH, or OCH3, or a dsose or osamine residue whose anomeric carbon in position 1 (C1) is engaged in a glycosidic bond, the positions C2, C3 and C4 being respectively substituted by groups R 1 2 R'3, and R'4 respectively having the meanings given above for R2, R3, and R4, and the position C6 by a group R 'having the meaning given above for R, at least one of R2, R'2, R3, R'3, R4 or R'4 representing a sulphate or phosphate residue.  11. Compounds according to claim 10 corresponding to the following formulas: α-β-methyl-glycoside N-acetyl-4-sulphate-6-OR2-desoxy-glucosamine, N-acetyl 4-sulphate-6-OR-2 desoxyglucosamine, α- or β-methylglycoside 4-Sulfate-6-OR-glucopyranose, 4-sulphate-6-OR-glucopyranose, 6,6-di-OR-4,4'-di-sulphate -a, a '-D-Trehalose, R having the meaning indicated in claim 9.  Compounds according to claim 10 or claim 11 corresponding to the following formulas: α- or β-methyl-glycoside N-acetyl-4-sulphate-6-octanoyl-2-deoxyglucosamine, α-or-p-methyl-glycoside-N acetyl-4-sulphate-6- (2,3-dipalmitoyl) glyceroyl-2-desoxy-glucosamine, N-acetyl-4-sulphate-6-octanoyl-2-desoxy-glucosamine, N-acetyl-4- Sulfate-6- (2,3-dipalmitoyl) -Glyceroyl-2-desoxy-D-glucosamine, α- or β-methyl-glycoside 4-Sulfate-6-octanoyl-glucopyranoside, α- or β-methyl-glycoside Sulphate-6- (2,3-dipalmitoyl) -glyceroyl-D-glucopyranoside, 4-sulphate-6-octanoyl-D-glucopyranose, 4-sulphate-6 (2,3-di-palmitoyl) -glyceroyl glucopyrano 4,4'-6,6-di-octanoyl-α, α'-D-Trehalose, 4,4'di-sulphate-6,6 '(2,3-dipalmitoyl) -glyceroyl-aa ' D-Trehalose.