WO1990006322A1

WO1990006322A1 - Peptidoglucan dimers, process for producing them and their use as immunomodulators

Info

Publication number: WO1990006322A1
Application number: PCT/EP1989/001454
Authority: WO
Inventors: Johanna Uschkoreit; Gerd Petrik
Original assignee: Helopharm W. Petrik Gmbh & Co., Kg
Priority date: 1988-12-01
Filing date: 1989-12-01
Publication date: 1990-06-14
Also published as: DE3840538A1; AU4655989A

Abstract

Peptidoglucan dimers of general formula (I), in which R?1, R2, R3 and R4¿ have the meaning given in claim 1. Also described is a process for producing these peptidoglucan dimers and immunomodulating agents containing these peptidoglucan dimers.

Description

"Peptidoglucan dimers, process for their preparation and their use as immunomodulators"

The invention relates to peptidoglucan dimers and immunomodulating agents which contain these peptidoglucan dimers.

Different immunodulators are known which influence non-specific immune reactions. These include bacterial cell walls and building blocks thereof, the tracking of which enables the cellular or humoral immune response to be increased unspecifically; see. Adam et al., Biochem. Bophys. Res. Comm. 56: 561 (1974), Ellouz et al. , Biochem. Biophys. Res. Com un. 59 (1974), 1317, Drews, Immunpharmako¬ logy, principles and perspectives, Springer-Verlag, Berlin 1986, pp 171 - 174. bacterial The ^'at least required for the immunomodulatory effect block cell walls is the N-acetyl-L-alanyl -D-isoglutamine (MDP for Muramyl peptide). Various chemical modifications and analogues of MDP have also already been produced, on the one hand to increase its immunomodulatory activity and on the other hand to reduce the side effects that occur when MDP is administered, such as pyrogenicity and athritogenicity, cf. e.g. Matsu oto et al., Inf. Immun. 39 (1983), 1029 - 1040. Experiments with MDP and the MDP modifications have shown that the immunomodulating effects of MDP depend on the balance of lipophilic and hydrophilic structures in the molecule. If the more lipophilic peptidyl portion in the aphipatic MDP structure is extended by aliphatic groups, this reduces the pyrogenicity and other side effects and leads to an extension of the in vivo half-life and to an increased affinity for the target cells.

Muramyl dipeptides, the monomers of which are connected by C ₁ -y-alkyl residues, are described in JP-A 55-28 933.

MDP can be used as an immunomodulator for infection prophylaxis, for combating bacterial and viral infections and for the treatment of tumor diseases and leukopenia after chemo and / or radiation therapy. However, the previously known MDP derivatives still have side effects which are too strong in relation to their immunomodulating activity, so that the application results achieved so far are not yet satisfactory.

The technical problem underlying the invention is therefore to provide MDP derivatives and analogs which have high immunomodulating activity and fewer side effects than those known hitherto.

The solution to this technical problem is achieved by providing the peptidoglucan dimers according to the invention which contain MDP, derivatives or analogues thereof as monomers which are linked by a C ₁₀ _ ₃₀ -α, ώ? -Diaminoalkylene radical .

The invention thus relates to peptidoglucan dimers of the general formula I:

R ¹ -R ² -R ³ -R ⁴ -R ³ -R ² -R ¹ (I) in which the leftovers

R ¹ , which can be the same or different, glucopyranose compounds derived from galacturon, mannuronic or glucuronic acid, or ribo-, arabino-, xylo- or lyxo-2-hexulopyranos-l-oyl-verbin ¬ dung or the corresponding 2-hexulofuranos-l-oyl compounds or N-acetylmuramic acid, N-acetylgalactosamic acid or N-acetylglucosamic acid, (1,2: 5,6-di-O-isopropylidene-D- mean glucofuranosyl-3-O) -acetic acid or (2-D-glucopyranosyl-3-O) -acetic acid, the above compounds optionally being provided with protective groups customary in the field of sugar chemistry and / or the 6-hydroxy group optionally being an acyl - rest with up to 90 carbon atoms, preferably a Smeg amycoloyl, Mycomycolyl, Corynomycoloyl, Mycoloyl, Nocardomycoloyl, 2-tetradecylhexadecanoyl, 3-Hydroxy-2-tetradecyloctadecanoyl-, 2-Docosyl-tetraco -, 3-Hydroxy-2-docosylhexacosanoyl-, isopentadeca- noyl-, acetyl-, butyryl-, octanoyl-, decanoyl-, Lauroy1-,

Myristoyl, palmitoyl, heptadecanoyl, stearoyl, non-decanoyl, eicosanoyl, tetracosanoyl or triacononoyl radical;

R ² , which may be the same or different, is a natural. A ionic acid, such as a glycine, L-alanine, L-arginine, L-asparagine, L-cysteine, L-glutamine, L-histidine, L-hydroxyproline, L-isoleucine, L-leucine, L-methionine, L-proline, L-serine, L-threonine or L-valine residue or a / 3-alanine residue or an N-methyl derivative of the above amino acids or L-α-aminobutyric acid , preferably a glycine, L-alanine, L-serine, L-valine or an L-α-aminobutyric acid residue and in particular a glycine or L-alanine residue;

R ³ , which may be the same or different, represent a D-glutamine, D-iso-glutamine or D-glutamic acid residue; R ^{4 is} a straight-chain or branched-chain α, 47-diaminoalkylene radical, the number of carbon atoms being at least 10, preferably 12, particularly preferably 14, in particular 16 and at most 30, preferably 25, particularly preferably 23, in particular 20 and in which Alkylene chain optionally up to 10 carbon atoms, preferably up to 8 carbon atoms, particularly preferably up to 6 carbon atoms, are substituted by nitrogen, oxygen and / or sulfur atoms, oxygen being preferred.

Particularly preferred as radical R ¹ is an acyl radical which is derived from N-acetylmuramic acid, N-acetylglactosamic acid, N-acetylglucosamic acid, glucuronic acid, galacturonic acid, mannuronic acid, (1,2: 5,6-di- 0-isόpropylidene-D-glucofuranosyl-3-O) -acetic acid, (2-D-glucopyranosyl-3-0) -acetic acid or (2,3: 4,6-di-0-isopropylidene-α: -L-xylo-2-hexulofuranose) -1-carboxylic acid.

Preferred protecting groups are: Formula Name

Acetyl

Benzoyl

Benzyl benzylidene (bound to two oxygen)

ethyl

Ethylidene (bound to two oxygen)

Isopropylidene

Methylsulfonyl methyl

Methylene (bound to two oxygen)

Nitro p-tolylsulfonyl

Trifluoromethylsulfonyl

-Tetrahydropyranyl Particularly preferred protecting groups are benzyl, benzylidene, acetyl and isopropylidene residues, especially isopropylidene residues. Preferred acyl radicals on the 6-hydroxy group of the radicals R ¹ are myσoloyl, myristoyl and stearoyl radicals.

According to the invention / 3 peptidoglucan dimers are preferred in which both radicals R ^{3 are} D-iso-glutamine radicals.

Examples of preferred monomers contained in the peptidoglucan dimers of the general formula I according to the invention

N-acetylmuramyl-L-alanyl-D-isoglutamine,

N-acetylmuramyl-L-valyl-D-isoglutamine,

N-acetylmuramyl-L-seryl-D-isoglutamine, N-acetylmuramyl-glycine-D-isoglutamine,

6-O-stearoyl-N-acetylmuramyl-L-alanyl-D-isoglutamine,

6-O-stearoyl-N-acetylmuramyl-L-valyl-D-isoglutamine,

6-O-stearoyl-N-acetylmuramyl-L-seryl-D-isoglutamine,

6-O-stearoyl-N-acetylmuramyl-glycine-D-isoglutamine, 6-O-mycoloyl-N-acetylmuramyl-L-alanyl-D-isoglutamine,

6-O-mycoloyl-N-acetylmuramyl-L-valyl-D-isoglutamine,

6-O-mycoloyl-N-acetylmuramyl-L-seryl-D-isoglutamine,

6-O-mycoloyl-N-acetylmuramyl-glycyl-D-isoglutamine,

N-acetylglucosaminyl-L-valyl-D-isoglutamine, N-acetylglucosaminyl-L-seryl-D-isoglutamine,

N-acetylglucosaminyl-glycyl-D-isoglutamine,

N-acetylglucosaminyl-L-alanyl-D-isoglutamine,

N-acetylgalactosaminyl-L-alanyl-D-isoglutamine,

N-acetylgalactosaminyl-L-valyl-D-isoglutamine, N-acetylgalactosaminyl-L-seryl-D-isoglutamine,

N-acetylgalactosaminyl-glycyl-D-isoglutamine,

(2,3: 4,6-di-0-isopropylidene-α-L-xylo-2-hexulofuranos-l-oyl) -

L-alanyl-D-isoglutamine,

(2,3: 4,6-di-0-isopropylidene-α-L-xylo-2-hexulofuranos-l-oyl) - L-seryl-D-isoglutamine,

(2,3: 4,6-di-0-isopropylidene-α-L-xylo-2-hexulofuranos-l-oyl) -

L-valyl-D-isoglutamine, (2,3: 4,6-di-0-isopropylidene-α-L-xylo-2-hexulofuranos-1-oyl) - glycyl-D-isoglutamine,

N- (1,2: 5,6-di-O-isopropylidene-D-glucofuranosyl-3-O-acetyl) -

L-alanyl-D-isoglutamine, N- (1,2: 5.6-di-0-isopropylidene-D-glucofuranosyl-3-0-acetyl) -

L-valyl-D-isoglutamine,

N- (1, 2: 5,6-di-0-isopropylidene-D-glucofuranosyl-3-0-acetyl) -

L-seryl-D-isoglutamine,

N- (1,2: 5,6-di-0-isopropylidene-D-glucofuranosyl-3-0-acetyl) glycyl-D-isoglutamine,

N- (2-D-glucopyranosyl-3-0-acetyl) -L-alanyl-D-isoglutamine,

N- (2-D-glucopyranosyl-3-0-acetyl) -L-valyl-D-isoglutamine,

N- (2-D-glucopyranosyl-3-0-acetyl) -L-seryl-D-isoglutamine,

N- (2-D-glucopyranosyl-3-0-acetyl) glycyl-D-isoglutamine,

N-acetylmuramyl-L-alanyl-D-glutamine,

N-acetylmuramyl-L-valyl-D-glutamine,

N-acetylmuramyl-L-seryl-D-glutamine,

N-acetylmuramylglycine-D-glutamine, 6-O-stearoyl-N-acetylmuramyl-L-alanyl-D-glutamine,

6-O-stearoyl-N-acetylmuramyl-L-valyl-D-glutamine,

6-O-stearoyl-N-acetylmuramyl-L-seryl-D-glutamine,

6-O-stearoyl-N-acetylmuramyl-glycine-D-glutamine,

6-O-mycoloyl-N-acetylmuramyl-L-alanyl-D-glutamine, 6-O-mycoloyl-N-acetylmuramyl-L-valyl-D-glutamine,

6-0-mycoloyl-N-acetylmuramyl-L-seryl-D-glutamine,

6-O-mycoloyl-N-acetylmuramyl-glycyl-D-glutamine,

N-acetylglucosaminyl-L-valyl-D-glutamine,

N-acetylglucosaminyl-L-seryl-D-glutamine, N-acetylglucosaminyl-glycyl-D-glutamine,

N-acetylglucosaminyl-L-alanyl-D-glutamine,

N-acetylgalactosaminyl-L-alanyl-D-glutamine,

N-acetylgalactosaminyl-L-valyl-D-glutamine,

N-acetylgalactosaminyl-L-seryl-D-glutamine, N-acetylgaϊactosaminyl-glycyl-D-glutamine,

(2,3: 4,6-di-0-isopropylidene-α-L-xylo-2-hexulofuranos-l-oyl)

L-alanyl-D-glutamine, (2, 3: 4, 6-di-0-isopropylidene-o: -L-xylo-2-hexulofuranos-l-oyl) -

L-seryl-D-glutamine,

(2, 3: 4, 6-di-0-isopropylidene-α-L-xylo-2-hexulo £ uranos-l-oyl) -

L-valyl-D-glutamine, (2, 3: 4, 6-di-0-isopropylidene-α-L-xylo-2-hexulofuranos-l-oyl) glycyl-D-glutamine,

N- (1,2: 5,6-di-O-isopropylidene-D-glucofuranosyl-3-O-acetyl) -

L-alanyl-D-glutamine,

N- (1,2: 5.6-di-0-isopropylidene-D-glucofuranosyl-3-0-acetyl) - L-valyl-D-glutamine,

N- (1,2: 5,6-di-O-isopropylidene-D-glucofuranosyl-3-O-acetyl) -

L-seryl-D-glutamine,

N- (1,2: 5,6-di-0-isopropylidene-D-glucofuranosyl-3-0-acetyl) - glycyl-D-glutamine, N- (2-D-glucopyranosyl-3-0-acetyl) - L-alanyl-D-glutamine,

N- (2-D-glucopyranosyl-3-0-acetyl) -L-valyl-D-glutamine,

N- (2-D-glucopyranosyl-3-0-acetyl) -L-seryl-D-glutamine,

N- (2-D-glucopyranosy1-3-O-acety1) glycyl-D-glutamine,

N-acetylmuramyl-L-alanyl-D-glutamic acid, N-acetylmuramyl-L-valyl-D-glutamic acid, N-acetylmuramyl-L-seryl-D-glutamic acid, N-acetylmuramyl-glycine-D-glutamic acid, 6-O- Stearoyl-N-acetylmuramyl-L-alanyl-D-glutamic acid, 6-O-Stearoy1-N-acetylmuramyl-L-valyl-D-glutamic acid, 6-0-stearoyl-N-acetylmuramyl-L-seryl-D-glutamic acid, 6-O-stearoyl-N-acetylmuramyl-glycine-D-glutamic acid, 6-0-mycoloyl-N-acetylmuramyl-L-alanyl-D-glutamic acid, 6-O-mycoloyl-N-acetylmuramyl-L-valyl-D- glutamic acid, 6-O-mycoloyl-N-acetylmuramyl-L-seryl-D-glutamic acid, 6-0-mycoloyl-N-acetylmuramyl-glycyl-D-glutamic acid, N-acetylglucosaminyl-L-valyl-D-glutamic acid, N- Acetylglucosaminyl-L-seryl-D-glutamic acid, N-acetylglucosaminyl-glycyl-D-glutamic acid, N-acetylglucosaminyl-L-alanyl-D-glutamic acid, N-acetylgalactosaminyl-L-alanyl-D-glutamic acid, N-acetylgalactyl valyl-D-glutamic acid, N-acetylgalactosaminyl-L-seryl-D-glutamic acid,

N-acetylgalactosaminyl-L-glycyl-D-glutamic acid,

(2,3: 4,6-di-0-isopropylidene-α-L-xylo-2-hexulofur-anos-l-oyl) -

L-alanyl-D-glutamic acid, (2,3: 4,6-di-0-isopropylidene-o_-L-xylo-2-hexulofuranos-l-oyl) -

L-seryl-D-glutamic acid,

(2,3: 4,6-di-0-isopropylidene-α-L-xylo-2-hexulofuranos-1-oyl) -

L-valyl-D-glutamic acid,

(2,3: 4,6-di-0-isopropylidene-α-L-xylo-2-hexulofuranos-l-oyl) - glycyl-D-glutamic acid,

N- (1,2: 5,6-di-0-isopropylidene-D-glucofuranosyl-3-0-acetyl) -

L-alanyl-D-glutamic acid,

N- (1,2: 5,6-di-0-isopropylidene-D-glucofuranosyl-3-0-acetyl) -

L-valyl-D-glutamic acid, N- (1,2: 5,6-di-0-isopropylidene-D-glucofuranosyl-3-0-acetyl) -

L-seryl-D-glutamic acid,

N- (1,2: 5,6-di-0-isopropylidene-D-glucofuranosyl-3-0-acetyl) glycyl-D-glutamic acid

N- (2-D-glucopyranosyl-3-0-acetyl) -L-alanyl-D-glutamic acid, N- (2-D-glucopyranosyl-3-0-acetyl) -L-valyl-D-glutamic acid,

N- (2-D-glucopyranosyl-3-0-acetyl) -L-seryl-D-glutamic acid,

N- (2-D-glucopyranosyl-3-O-acetyl) glycyl-D-glutamic acid.

The peptidoglucan dimers according to the invention consisting of alternating L- and D-amino acid residues, sugar residues and aliphatic building blocks are not degradable by conventional proteolytic enzymes. They also have improved biological activity over the monomers due to the doubling of the biologically active ingredient. They are therefore more suitable as medicinal substances than the corresponding monomers.

According to the invention, particularly preferred peptidoglucan dimers are N ^α , N ° bis - [(2,3: 4,6-di-0-isopropylidene-α-L-xylo-2-hexulofuranos-1-oyl -) - L-alanyl -D-isoglutaminyl)] -diaminode- can, N ^α , N -Bis- (N-acetylmuramyl-L-alanyl-D-isoglutaminyl) - diaminodecane, N ^α , N ^G5 -Bis- (N-acetylmuramyl-L-alanyl -D- isoglutaminyl) -diaminooctadecan, N ^α , N -Bis- (N-acetylmuramyl-L-alanyl-D-isoglutaminyl) -diaminopentaethylene glycol and N ^α - (N-acetylmuramyl-L-alanyl-D-isoglutaminyl) -N ^ (6- O-mycoloyl-N-acetylmuramyl-L-alanyl-D-isoglutaminyl) diaminodecane. The peptidoglucan dimers according to the invention can be prepared by first synthesizing the monomers in a manner known per se and then combining them in a manner known per se together with the α, -diaminoalkylene radical to give dimers. The linkage reaction can be described, for example, by Koenig et al., Chem. Ber. 103 (1970), 2024-2033.

Pyran derivatives can be prepared by first synthesizing the corresponding .. furan derivatives and then converting them into the pyran derivatives in a manner known per se; see. Chikashita et al., J. Heterocyclic Chem. 19 (1982), 981.

The invention further relates to immunomodulating agents or medicaments which contain at least one of the peptidoglucan dimers according to the invention. These agents can be used for the prophylaxis of infections, for the control of bacterial and viral infections and for the treatment of tumor diseases and leukopenia after Che o- and / or radiation therapy. These agents can also be used as adjuvants.

The agents or medicaments according to the invention are preferably tablets or gelatin capsules which contain the peptidoglucan dimers together with diluents, such as lactose, dextrose, sucrose, mannitol, sorbitol or cellulose, and / or lubricants, such as silica, tallow, stearic acid or salts of which, such as magnesium or calcium stearate, and / or contain polyethylene glycol. Tablets may contain binders such as magnesium aluminum silicate, starches such as corn, wheat, rice or arrowroot starch, gelatin, tragacanth, methyl cellulose, sodium carboxymethyl cellulose cellulose and / or polyvinylpyrrolidone and optionally also disintegrants, such as starches, agar, alginic acid or a salt thereof, such as sodium alginate, and / or effervescent mixtures, or adsorbents, colors, flavors and sweeteners. Suppositories, ointments or creams are mainly fat emulsions or suspensions. “The medicaments according to the invention can be sterilized and / or contain auxiliaries, for example preservatives, stabilizers, wetting agents and / or emulsifiers, solubilizers, salts for regulating the osmotic pressure and / or buffers. These pharmaceutical preparations, which may optionally contain further pharmaceutically valuable substances, are produced in a manner known per se, for example by conventional mixing, granulating or coating processes, and contain about 0.1 to about 75 %, in particular about 1 to 50%, of the peptidoglucan dimers according to the invention.

In addition, the peptidoglucan dimers according to the invention can be administered with the aid of conventional liposomes.

The examples illustrate the invention.

example 1

Preparation of N ^α , N ^ -Bis- [N-acetylmuramyl-L-alanyl-D-isoglutaminyl] diaminodecane

Method A

a) Preparation of N ^α , N ° -Bis- [L-alanyl-D-iso-glutaminyl] - diaminodecane

344.6 mg (2 mmol) of diaminodecane are dissolved in 10 ml of dimethylformamide in the presence of 200 μl of triethylamine. 0.5 g (2.4 mmol) of dicyclohexylcarbodiimide (DCC) and 0.32 g (2.4 mmol) of hydroxybenzotriazole (HOBT) are added to the solution. After the reactants are completely dissolved, that becomes The reaction mixture was cooled to -15 ° C. and added dropwise within 72 hours with 1.7 g (5 mmol) of ZL-Ala-D-Glu-NH ₂ (Z = benzyloxycarbonyl) in 5 ml of dimethylformamide / dimethyl sulfoxide in a ratio of 1 : 1 offset. The reaction mixture is then left to stand at room temperature for 3 hours and the precipitated dicyclohexylurea is filtered off. The reaction mixture is concentrated to 6 ml under reduced pressure and left to stand for 48 hours at room temperature. The protective group Z is removed by a known method. Thin-layer chromatography (silica gel; butanol / acetic acid / H ₂ 0 in a volume ratio of 4: 2: 2 or isopropanol / H ₂ 0 in a volume ratio of 7: 3) shows that the coupling reaction has proceeded quantitatively.

b) Preparation of N ^α , N ⁵ -Bis- [N-acetylmuramyl-L-alanyl-D-isoglutaminyl) diaminodecane

5 g (10.27 mmol) of N-acetyl-α-benzyl-4,6-0-benzylidene-muraminic acid are dissolved in 10 ml of dimethylformamide and with 2 g (10 mmol) of dicyclohexylcarbodiimide and 1.5 g (10 mmol ) Hydroxybenzotriazole added. The reaction mixture is left to stand at room temperature for 3 hours. The precipitated dicyclohexylurea is filtered off and the activated carboxyl compound is added dropwise to the amino compound obtained in a). Dicyclohexylurea precipitates again, which is filtered off. The total volume of the reaction mixture is then concentrated to 10 ml under reduced pressure and left to stand for 48 hours at room temperature. After concentration again, the residue is dissolved in tetrahydrofuran (200 ml) and hydrogenated in the presence of palladium black at room temperature. The title compound is obtained by HPLC.

Method B

Preparation of N ^α , N ^ö -Bis- [N-acetylmuramyl-L-alanyl-D-isoglutaminyl] diaminodecane 2 g (4.1 mmol) of N-acetyl-α-benzyl-4,6-O-benzylidene-muraminic acid are dissolved in 10 ml of dimethylformamide and mixed with 1 g (4.85 mmol) of dicyclohexylcarbodiimide and 600 mg (4, 4 mmol) of hydroxybenzotriazole were added. Dicyclohexylurea forms in the reaction mixture, which is filtered off after about 1 hour. As indicated in method A, the reaction mixture is cooled to -15 ° C. and then 868 mg (4 mmol) of HL-Ala-D-isoglutamine in 7 ml of diethyl formamide are added in portions within 7 hours. The reaction mixture is left to stand at room temperature for 1 day and the dimethylformamide is concentrated to a residual volume of 6 ml under reduced pressure. 344.6 mg (2 mmol) of diaminodecane, 824 mg. (4 mmol) dicyclohexylcarbodiimide and 540 mg (4 mmol) hydroxybenzotriazole were added. The reaction mixture is then left to stand at room temperature for 2 days. The precipitated dicyclohexylurea is filtered off and the solvent is evaporated off under reduced pressure. It leaves a syrupy residue. After hydrogenolysis as in method A, the title compound is obtained by means of HPLC.

Example 2

Preparation of N ^α , KT-bis - [(2,3: 4,6-di-0-isopropylidene-o: -L-xylo-2-hexulofuranos-l-oyl) -L-alanyl-D-isoglutaminyl] - diaminodecane

Method A

a) ^Preparation of N ^α , N ^ÄJ -Bis- [L-al.anyl-D-iso-glutaminyl] - diaminodecane

The title compound is prepared as described in Example 1, Method A a).

b) Representation of N ^α , N ^ω -Bis - [(2,3: 4,6-di-0-isopropylidene-α-L-xylo-2-hexulofuranos-l-oyl) -L-alanyl-D-isoglutami - nyl)] diaminodecane

3 g (10.27 mmol) of 2,3: 4,6-di-θ-isopropylidene-2-keto-L-gulonic acid are dissolved in 10 ml of dimethylformamide and with 2 g (io mmol) of dicyclohexylcarbodiimide and 1.5 g (10 mmol) of hydroxybenzotriazole were added. The reaction mixture is left to stand at room temperature for 3 hours. The precipitated dicyclohexylurea is filtered off and the activated carboxyl compound is added dropwise to the amino compound obtained in a). Dicyclohexylurea precipitates again, which is filtered off. The total volume of the reaction mixture is then concentrated to 10 ml under reduced pressure and left to stand for 48 hours at room temperature. The title compound is isolated by HPLC.

Method B

N ^α , N ^ bis - [(2,3: 4,6-di-0-isopropylidene-α-L-xylo-2-hexulofuranos-1-oyl) -L-alanyl-D-isoglutaminyl)] - diaminodecane

1 2 g, (4.1 mmol) of diacetone-2-keto-L-gulonic acid are dissolved in 10 ml of dimethylformamide and 1 g (4.85 mmol) of dicyclohexylcarbodiimide and 600 mg (4.4 mmol) of hydroxybenzotriazole are added. Dicyclohexylurea is formed in the reac. tion mixture, which is filtered off after about 1 hour. As indicated in Example 1, the reaction mixture is cooled to -15 ° C. and then 868 mg (4 mmol) of HL-Ala-D-isoglutamine in 7 ml of dimethylformamide are added in portions within 7 hours. The reaction mixture is left to stand at room temperature for 1 day and the dimethylformamide is removed under reduced pressure, so that a residual volume of 6 ml remains. 344.6 mg (2 mmol) of diaminodecane, 824 mg (4 mmol) of dicyclohexylcarbodiimide and 540 mg (4 mmol) of hydroxybenzotriazole are added. The reaction mixture is then left to stand at room temperature for 2 days. The precipitated dicyclohexylurea is filtered off and the solution evaporated medium under reduced pressure. The following remains: a syrup-like residue which is analyzed by thin-layer chromatography. The title compound is obtained from this by HPLC.

Example 3

Preparation of N ^α , N -Bis- [N-acetylmuramyl-L-alanyl-D-isoglutaminyl] diaminooctadecane

The title compound is prepared as indicated in Example 1.

Example 4

Preparation of N ^ N ^ -Bis-fN-acetylmuramyl-L-alanyl-D-isoglutaminyl] -diaminopentaethylene glycol

4.8 g (10.27 mmol) of N-acetyl-α: -benzyl-4,6-O-benzylidene-muramic acid are dissolved in 10 ml of dimethylformamide and with 2 g (10 mmol) of dicyclohexylcarbodiimide and 1.5 g (10 mmol) of hydroxybenzotriazole added. Dicyclohexylurea forms in the reaction mixture, which is filtered off after about 1 hour. The reaction mixture is cooled to -10 ° C. and 868 mg (4 mmol) of HL-Ala-D-isoglutamine in 7 ml of dimethylformamide are then added in portions within 7 hours. The reaction mixture is left to stand at room temperature for 1 day and the dimethylformamide is concentrated to a residual volume of 6 ml under reduced pressure. There are 472 mg (7 mmol) of α, ^* ü-diaminopentaethylene glycol (H ₂ N- (CH ₂ -CH ₂ -0) ₄ -CH ₂ -CH ₂ -NH ₂ ), 824 mg (4 mmol) of dicyclohexylcarbodiimide and 540 mg (4 mmol) hydroxybenzotriazole added. The reaction mixture is then left to stand at room temperature for 2 days. The precipitated dicyclohexylurea is filtered off and the solvent is evaporated off under reduced pressure. It leaves a syrup residue. After hydrogenation analogously to Example 1 Method A, the title compound is obtained by means of HPLC.

Example 5

Representation of N ^α - (N-acetylmuramyl-L-alanyl-D-isoglutamininyl-N ⁰⁰ - (6-O-mycoloyl-N-acetylmuramyl-L-alanyl-D-isoglutamininyl) -diaminodecane

a) Preparation of N ^α -Fmoc-diaminodecane

To produce N ^α -Fmoc-diaminodecane (Fmoc. = Fluorenylethyloxycarbonyl-), 1 equivalent of α, <U-diaminodecane is reacted with 0.7 equivalents of Fmoc-Cl in 5% NaHC0 ₃ solution and tetrahydrofuran for 4 hours. The solution is concentrated and the product is obtained by means of chromatography on silica gel.

b) Preparation of l-benzyl-6-O-mycoloyl-N-acetyl-muramyl-L-alanyl-D-isoglutamine

The title compound is prepared analogously to S. Kusumoto, S. Okada and T. Shiba, Tetrahedron Letters 1976, 4287.

c) Preparation of N ^α - (N-acetyl-1 ^α -benzyl-4, 6-O-benzylidene-muramyl-L-alanyl-D-isoglutaminyl) diaminodecane

• N-acetyl-l r ^u r-benzyl-4,6-O-benzylιden-muramyl-L-alanyl-D-glutamine ιso- is reacted with N ^α -Fmoc-diaminodecane as follows: 2.8 g of (4 1 mmol) of N-acetyl-1 ^α -benzyl-4,6-0-benzylidene-muramyl-L-alanyl-D-isoglutamine are mixed with 1.57 g (4 mmol) of N ^α -Fmoc-diaminodecane, 824 mg (4 mmol) of dicyclohexylcarbodiimide and 540 mg (4 mmol) of hydroxybenzotriazole dissolved in 10 ml of dimethylformamide.

The reaction mixture is then left to stand at room temperature for 2 days. The precipitated dicyclohexylurea is filtered off and the solvent evaporated under reduced pressure. The Fmoc group is removed using piperidine using a known method and the product is purified by HPLC.

d) Preparation of N ^α - (N-acetylmuramyl-L-alanyl-D-isoglutaminyl-IT ³ - (6-O-mycoloyl-N-acetylmuramyl-L-alanyl-D-isoglytaminyl) diaminodecane

2 mmol each of compound b) and compound c) are dissolved in tetrahydrofuran and coupled in the presence of equimolar amounts of DCC / HOBT overnight at -10 ° C. and at room temperature for two more days. After removal of the dicyclohexylurea, the solvent is reduced under reduced pressure The protective group is removed by hydrogenation (see ref. In b) and the title compound is isolated by means of HPLC.

Claims

Patent claims

1. Peptidoglucan dimers of the general formula I:

R ¹ -R ² -R ³ -R ⁴ -R ³ -R ² -R ¹ (I)

in which the leftovers

R ¹ , which may be the same or different, glucopyranose compounds which differ from the galacturon. Derive mannuronic or glucuronic acid, or ribo-, arabino-, xylo- or lyxo-2-hexulopyranos-1-oyl compounds or the corresponding 2-hexoxofuranos-1-oyl compounds or N-acetylmuramic acid, N-acetylgalactosamic acid or N-acetylglucosamic acid, (1,2: 5,6-di-O-isopropylidene-D-glucofuranosyl-3-0) -acetic acid or (2-D-glucopyranosyl-3-0) - mean acetic acid, where the above compounds are optionally provided with protective groups customary in the field of sugar chemistry and / or the 6-hydroxy group optionally carries an acyl radical with up to 90 carbon atoms; R, which may be the same or different, a natural amino acid, such as a glycine, L-alanine, L-arginine, L-asparagine, L-cysteine, L-glutamine, L-histidine, L-hydroxyproline, L-isoleucine, L-leucine, L-methionine, L-proline, L-serine, L-threonine or L-valine residue or a ß-alanine residue or an N-methyl derivative above amino acids or L-α-aminobutyric acid; R ³ , which may be the same or different, have a D

Mean glutamine, D-iso-glutamine or D-glutamic acid residue;

R ^{4 is} a straight-chain or branched-chain α, ö-

Diaminoalkylenerest having 10 to 30 carbon atoms.

2. Peptidoglucan dimer according to claim 1, in which R ⁴ is substituted by oxygen atoms.

3. peptidoglucan dimer according to claim 1 or 2, wherein the number of carbon atoms is at most 25.

4. peptidoglucan dimer according to one of claims 1 to 3, in which R ¹ N-acetylmuramic acid, N-acetylgalactosamic acid, N-acetylglucosamic acid, glucuronic acid, galacturonic acid, mannuronic acid, (1,2: 5,6-di-O-isopropylidene) D-glucofuranosyl-3-0) -acetic acid, (2-D-glucopyranosyl-3-0) -acetic acid or (2,3: 4,6-di-0-isopropylidene-α: -L-xylo -2-hexulofuranose) -1-carboxylic acid.

5. Peptidoglucan dimer according to one of claims 1 to 4, in which the protective group is a benzyl, benzylidene, acetyl or isopropylidene radical.

6. Peptidoglucan dimer according to one of claims 1 to 5, in which the acyl radical on the 6-hydroxy group is a mycoloyl, myristoyl or stearoyl radical.

7. Peptidoglucan dimer according to one of claims 1 to 6, in ddeemm RR eeiinn GGllyycciinn-- ,, LL - AAllaanniinn-- ,, is L-serine, L-valine or an L-α-aminobutyric acid residue.

8. peptidoglucan dimer according to any one of claims 1 to 7, in which R ^{3 is} a D-iso-glutamine residue.

9. peptidoglucan dimer according to one of claims 1 to 8, characterized in that it is N ^α , N -Bis (N-acetylmura-myl-L-alanyl-D-isoglutaminyl) diaminodecane.

10. peptidoglucan dimer according to one of claims 1 to 8, characterized in that it is N ^α , N ^α; -Bis - [(2,3: 4,6-di-0-isopropylidene-α-L-xylo-2-hexulofuranos-1-oyl) -L-alanyl-D-isoglutaminyl] diaminodecane.

11. Peptidoglucan dimer according to one of claims 1 to 8, characterized in that it is N ^α , N ^ω- bis (N-acetylmura-myl-L-alanyl-D-isoglutaminyl) diaminooctadecane.

12. Peptidoglucan dimer according to one of claims 1 to 8, characterized in that it is N ^α , N ^CJ- bis (N-acetylmura-myl-L-alanyl-D-isoglutaminyl) -dia inopentaethylene glycol.

13. Peptidoglucan dimer according to one of claims 1 to 8, characterized in that it is N ^α - (N-acetylmuramyl-L-alanyl-D-isoglutaminyl) -N ^w - (6-O-mycoloyl-N-acetylmuramyl -L-alanyl-D-isoglutaminyl) diaminodecane.

14. A process for the preparation of the peptidoglucan dimers according to one of claims 1 to 13, in which the monomers are first prepared in a manner known per se and these are linked together in a manner known per se to form dimers together with the α C'-diaminoalkylene radicals.

15. Immunomodulating agent containing at least one of the peptidoglucan dimers according to one of claims 1 to 13.

16. A method for modulating the immune system of mammals, in which a patient who needs this modulation of the immune system is provided with an amount of a peptidoglucan dimer according to one of claims 1 to 13 which is sufficient for modulating the immune system, optionally together with pharmaceutically compatible carriers and additives.