DE2655500C2 - - Google Patents

Description

The invention relates to muramyl peptide derivatives, processes for the preparation of these compounds, and their use for Defense of the physical defense.

In J. Med. Chem. 9, 971-973 (1966) are without specific information use some N-acetyl-muramic acid derivatives described that are not within the scope of the present Claims fall. German Offenlegungsschrift 24 50 355 describes vaccines containing muramic acid derivatives, including MDP, with 2 to 8 amino acids in the peptide chain, which are not the subject of the present application belong. From Z. Immun. Forsch. 149, 341-348 (1975), Biochem. Biophys. Res. Comm. 59: 1317-1325 (1974) and ibid. 66, 1316-1322 (1975) is the compound N-acetyl-muramyl-L- alanyl-D-isoglutamine (abbreviated: MDP) known which also not within the scope of the present claims falls. This compound can also be called 2-acetamido-3-O- {l- D- [l-L- (1-D-carbamoyl-3-carboxypropyl) carbamoylethyl] - carbamoylethyl} -2-deoxy-D-glucose. The structure of MDP is part of one in nature, namely structure found in bacterial cell walls.  

In the last three literature references mentioned noted that MDP is the smallest adjuvant active Fragment of the natural structure. The state of the Technology thus suggests the expectation that all components of MDP are essential for the biological effect. This impression is given by Chem. Abstr. 83, 161,981 (1975), and the reference cited therein, S. Kotani et al., Biken J. 18 (2), 105-111 (1975), reinforced, from which shows that z. B. neither N-acetyl-muramyl-L-alanyl-L- isoglutamine or N-acetyl-muramyl-L-alanyl-D-isoasparagin are active in adjuvants. The latter finding confirmed only the expectations at the time of priority the professional world. It is common to peptides announced that the exchange of a single Amino acid in the sequence essential for the effect against another amino acid usually the biological one Effect changed profoundly. It is also of pharmacological active substances known that the absolute Configuration at existing asymmetry centers for the biological effect is essential, like that above mentioned example of the replacement of D-isoglutamine in the MDP by L-isoglutamine shows what leads to the loss of adjuvant activity leads. As from German Offenlegungsschrift 26 45 610 results in the replacement of L-alanine in the MDP D-Alanine for immunosuppressive properties. So it was also unpredictable what impact the change will have  the absolute configuration at one of the 8 asymmetry centers of MDP or the loss of an asymmetry center would have.

The object of the present invention was to be synthetic relatively easy to manufacture immunopotentiating To provide sugar derivatives that are better are tolerated as MDP and / or an increased immune potentiation cause.

The invention relates to general glucosamine derivatives formula

wherein RC _1-4 alkyl or phenyl, R₂ hydrogen or C _1-4 alkyl, R₇ hydrogen, C _1-4 alkyl, hydroxymethyl, mercaptomethyl or phenyl and R₈ and R₉ independently carboxyl, (C _1-4 alkoxy ) -carbonyl, carbamoyl or N-methyl-carbamoyl, with the proviso that the alkyl radical R has more than 1 carbon atom if the radical R₂ is methyl, or if the radical R₂ is hydrogen and R₈ and R₉ each represent a carboxyl group, and their salts.

Modified below with the expression "down" Contain residues, radicals or compounds, if not specified, preferably up to 7, primarily up to 4 carbon atoms.

C _1-4 alkyl is e.g. B. iso-propyl, straight-chain or branched, butyl bound in any position, and especially methyl, ethyl or n-propyl.

The lower alkylene dioxy group is in particular the To name methylenedioxy group.

Hologists are e.g. B. fluorine, chlorine or bromine. As C _1-4 alkoxycarbonyl groups in particular methoxycarbonyl or ethoxycarbonyl, but also n-propoxycarbonyl or isopropoxycarbonyl groups are to be mentioned and as carbamoyl groups in particular the carbamoyl group itself.

In the above compounds in which R₂ is a Means alkyl radical, is the one with the oxygen atom in 3- Position of the glucosamine residue linked R₂-acetic acid residue optically active, d. H. it is in D-shape. If R₇ is not Represents hydrogen, the R₇-aminoacetic acid is in L-form in front.

The new connections present are dependent on the nature of their substituents neutral or acidic compounds. If acidic groups are present, the salts also form Bases, such as ammonium salts or salts with alkali or alkaline earth metals, e.g. As sodium, potassium, calcium or magnesium.  

Pharmaceuticals are used only as pharmaceuticals usable, non-toxic salts.

The compounds of the present invention have valuable pharmacological properties, especially one pronounced immunopotentiating effect. This can be done on hand the experimental setup described below:

1. Potentiation of cellular immunity in vivo: Increase in late-type hypersensitivity to ovalbumin in guinea pigs.

Pirbright guinea pigs are given 10 mg on day 0 Ovalbumin in complete Freund's adjuvant by injection of 0.1 ml of an antigen-adjuvant mixture in each of the two Hind paws immunized. 4 weeks later there are skin reactions by intracutaneous injection of 100 µg ovalbumin in 0.1 ml buffered physiological saline solution and triggered the 24 hours later based on the erythema area and the Skin thickness increase quantified calculated reaction volume. The antigen-specific observed after 24 hours (late-type reaction)  Increase in the reaction volume is considered a Measure of cell-mediated immunity. Ovalbumin is one too weak immunogen, by itself or in a water oil emulsion with incomplete Freund's adjuvant (10 parts Ovalbumin solution in 0.9% NaCl mixed with 8.5 parts bayol F and 1.5 parts Arlacel A) to induce a late type reaction, but must be complete for effective immunization Adjuvant, mycobacteria are added (5 mg killed and lyophilized M butyricum per 10 ml Bayol® F / Arlacel® A) can be applied. For the detection of immunopotentiating The effects of test substances can now take the place the mycobacteria in doses of 10 to 100 µg the antigen-oil mixture be added.

The glucosamine peptides according to the invention are in capable of the effect of mycobacteria described in the Imitating the experimental set-up and surpassing it quantitatively.

A significant potentiation of late-type reactivity against ovalbumin can also be achieved in that Compounds of the type described are not in the antigen-oil mixture incorporated, but in doses of 10 to 100 µg each Animal for a few days after immunization (e.g. a day 0, 1, 2, 5, 6 and 7) administered subcutaneously in saline will.  

This shows that compounds of the described Are able to significantly increase cellular immunity, both in a mixture with the antigen itself (adjuvant effect in the narrower sense), as well as in terms of time and location separate supply from the antigen injection (systemic Immunopotentiation).

2. Potentiation of humoral immunity in vivo: increase of antibody production against bovine serum albumin (BSA) in the mouse.

NMRI mice are identified by intraperitoneal (i.p.) Injection of 10 µg of precipitate-free BSA immunized on day 0. 9, 15 and 29 days later, serum samples are taken and on their content of anti-BSA antibodies with a passive Hemagglutination technique examined. In the dose used soluble BSA is subimmunogenic for the recipient animals, d. H. it is capable of little or no production of antibodies. Additional treatment of the Mice with certain immunopotentiating substances before or after antigen administration leads to an increase in antibody titers in serum. The effect of the treatment is through the score achieved, d. H. by the sum of the log₂ titer differences expressed on the three bleeding days.  

Compound of the present invention are in the Position, with intraperitoneal or subcutaneous (see c.) Application from 100-300 mg / kg / animal on five consecutive days (Day 0 to 4) antibody production after immunization with BSA against BSA significantly.

The immune-stimulating effect of the compounds mentioned is in contrast to that of other bacterial immunoleptics (e.g. LPS from E. coli) antigen dependent: injection of the new compounds has only been immunized in BSA, not but in non-immunized mice an increase in the anti BSA titer result. Remarkably, the s. c. Gift of the compounds just as effective as the i. p. Application, d. that is, the immunopotentiating effect observed systemic and does not depend on the stimulant via the same route as the antigen or with the antigen must be administered mixed, as in classic Adjuvants is the case.

The experiments described show that Compounds of the type described also humoral immunity are able to specifically increase that they are immunological Improve stimulus response, and that its immunopotentiating Effects on systemic activation of the immune system are based.  

3. Potentiation of humoral immunity in vitro: T- Cell-substituting effect on the antibody response of Mouse spleen cells against sheep erythrocytes (SE).

For inducing an antibody response in many cases lymphocytes (T cells) from the thymus needed. These cells cooperate with the precursors antibody-forming lymphocytes (B cells) and help them on stimulation with so-called T-dependent antigens To respond to proliferation, differentiation and antibody synthesis. Spleen cell suspensions congenitally athymic nu / nu Mice contain no functional T cells and are capable of e.g. B. in vitro in the presence of SE no anti-SE antibodies to build. The compounds of the present invention are Surprisingly able to take T cells in such cultures functionally replace and an antibody response against SE to enable. Addition of these substances to nu / nu spleen cell cultures in the presence of SE leads to a significant within 4 days Increase in the number of antibody-producing cells. The findings show that the compounds mentioned humoral antibody formation increase in vitro and a defect of the T- Zell Systems are able to compensate.  

4. Selective mitogenicity for B cells: Proliferation-promoting Effect in B-lymphocyte cultures.

Suspensions of highly enriched B lymphocytes (Lymph node cells of congenitally athymic nu / nu mice), as well largely pure immature and mature T lymphocytes (Thymus cells or cortisone resistant, i.e. 48 hours after a cortisone injection of persistent Balb / c- thymus cells Mice) are in the presence of the test substances for three days incubated. The incorporation of H³-thymidine into the lymphocytes applies during the last 18 hours of the culture period as a measure of proliferation activity.

The compounds according to the invention are for B- Lymphocytes (i.e. for the precursors of antibody-producing Cells, but not mitogenic for T lymphocytes.

You are thus able to prevent the proliferation of To stimulate lymphocytes involved in the humoral immune response are.

compatibility

Although compounds of the type described are potentiating Effect on guinea pigs, for example after a single dose of 0.05 mg / kg s. c., on the mouse after  Application of 5 times 10 mg / kg s. c. unfold, too with application of 5 times 300 mg / kg i. p. no toxic to mice Effects observed. The substances mentioned have therefore over an excellent therapeutic range. The properties of the structurally most closely related connection from the prior art, d. H. from the above Compound N-acetyl-muramyl-L-alanyl-D-isoglutamine (MDP), are in DE-OS 27 18 010 with Compounds compared, some of which are also in the present Registration are described. In Table I in Example 16 correspond to that DE-OS:

Compound 1 on p. 205 of the previously known comparative compound MDP,
Compound 1 on p. 206 the end product of Example 3,
Compound 2 on p. 206 the end product of Example 1,
Compound 3 on p. 206 the end product of Example 26,
Compound 4 on p. 206 the end product of Example 27,
Compound 9 on p. 206 the end product of Example 5, and
Compound 1 on p. 207 the end product of Example 2.

The table shows that all of the above. Examples described Connections of the prior art Comparative connection are superior. Especially clear this superiority occurs when comparing the undesirable Side effects. For example, derives from Example 17B., pages 210 and 211 of the above. DE-OS, that cats at a dose of 1500 mg at the state of the Technology-related comparison with vomiting,  Diarrhea, depression, poor appetite and high fever responded while given the same dose of none of the substances described in Example 1 Disorders of well-being have been identified.

The article by K. Kamisango et al., Chem. Pharm. Bull. 29, 1644-1654 (1981) proves (see in particular the Summary and Table IV) that the glutamic acid derivative preceding amino acid of the MDP, d. H. Alanine, by numerous other amino acids can be replaced without the biological effect decreases.

The article by P. Lefrancier et al., J. Med. Chem. 25, 87-90 (1982), cf. Tables II and I in particular on page 88 show that the muramylpeptide C _1-4 alkyl esters are superior to the comparative compound MDP which belongs to the prior art in terms of adjuvant activity and, moreover, have significantly fewer undesirable side effects, as shown by the example of pyrogenicity becomes.

The compounds according to the invention have the ability on the one hand, when mixed with an antigen, its immunogenicity to increase, on the other hand with systemic application the immunological reactivity of the treated organism increase. The substances mentioned are able to both to promote cellular as well as humoral immunity and the lymphocytes responsible for antibody formation activate.  

The new compounds can thus be used as adjuvants in Mixture with vaccines can be used to increase vaccination success to improve and by humoral antibodies and / or cellular Immunity mediated infection protection against bacterial, improve viral or parasitic pathogens.

Finally, the compounds described are suitable in admixture with various antigens as adjuvants in the experimental and industrial production of antisera for therapy and diagnostics and for the induction of immunologically activated lymphocyte populations for cell transfer processes.

In addition, the new connections can also can be used without simultaneous antigen supply subliminal immune reactions in humans and animals to promote. The compounds are therefore particularly suitable for the stimulation of the body's defense, e.g. B. in chronic and acute infections or for selective (antigen-specific) immunological defects, as well as in congenital, however also for acquired general (i.e. non-antigen-specific) immunological defects, such as those in old age, in Course of severe primary diseases and especially after therapy with ionizing rays or with immunosuppressive ones Hormones occur. The substances mentioned can thus preferably also in combination with anti-infectious antibiotics, Chemotherapeutic agents or other healing methods are administered  to counteract immunological damage. After all, they are described substances also for the general prophylaxis of infectious diseases suitable for humans and animals.

Compounds of the formula I are particularly valuable, in which R₂ represents hydrogen and the other radicals have the above meaning, and their salts.

Particularly noteworthy are compounds of the general formula I, in which RC _1-4 alkyl or phenyl, R₂ is hydrogen or methyl, R₇ is hydrogen, C _1-4 alkyl or hydroxymethyl, R₈ carbamoyl and R₉ are carboxyl, with the proviso that the lower alkyl radical R has more than 1 carbon atom if R₂ is methyl, and their salts.

The invention also relates to the following methods for the preparation of the compounds of the general Formula I.

The new compounds can be obtained if a compound of the formula is known per se

wherein R and R₂ have the meaning given above and R₁ °, R₄ ° and R₆ ° stand for an easily removable protective group, or a derivative thereof with a compound of the formula

wherein R₇ °, R₈ ° and R₉ ° have the meaning of R₇, R₈ and R₉, with the proviso that carboxyl present in these residues and, if desired, free hydroxyl groups by easily removable Protecting groups are protected, condensed and, if necessary splits off existing protecting groups.

The condensation takes place z. B. in the way that the compound III in the form of the activated carboxylic acid with the amino compound IV or that the Acid III with the compound IV, whose amino group in activated Form is present, implemented. The activated carboxyl group For example, an acid anhydride, preferably a mixed one Acid anhydride like an acid azide, an acid amide like an imidazolide, isoxazolide or an activated ester. As activated esters may be mentioned in particular: cyanomethyl ester, Carboxymethyl ester, p-nitrophenyl thioester, p-nitrophenyl ester, 2,4,5-trichlorophenyl ester, pentachlorophenyl ester, N-hydroxy succinimide ester, N-hydroxyphthalimide ester, 8-hydroxyquinoline ester, 2-hydroxy-1,2-dihydro-1-carboethoxy-quinoline ester, N- Hydroxypiperidine esters or enol esters with N-ethyl-5-phenyl  Isoxazolium-3'-sulfonate can be obtained. Activated esters can also optionally with a carbodiimide with the addition of N- Hydroxysuccinimide or an unsubstituted or z. B. by Halogen, methyl or methoxy substituted 1-hydroxybenzotriazole, 3-Hydroxy-4-oxo-3,4-dihydro-benzo [d] -1,2,3-triazine obtained will.

The amino group is, for example, by reaction activated with a phosphitamide.

Among the methods of reaction with activated esters are especially those with N-ethyl-5-phenyl-isoxazolium-3'- sulfonate (Woodward Reagens K) or 2-ethoxy-1,2-dihydro-1- carboethoxy-quinoline or carbodiimide to mention.

Easily removable protective groups are those that are known from peptide or sugar chemistry. For carboxyl groups are especially tertiary-butyl, benzyl or benzhydryl and for hydroxyl groups in particular acyl radicals, e.g. B. Lower alkanoyl residues such as acetyl, aroyl residues such as benzoyl and especially carbon-derived residues such as benzyloxycarbonyl or lower alkoxycarbonyl, or alkyl, in particular tert. Butyl, optionally by nitro, lower alkoxy or Halogen substituted benzyl or tetrahydropyranyl or optionally substituted alkylidene residues containing the oxygen atoms connect in 4- and 6-position. Such Alkylidene radicals are in particular a lower alkylidene, in the first Line of the ethylene, isopropylidene or propylidene residue or  also an optionally substituted, preferably in the p-position substituted, benzylidene radical.

These protective groups can be known per se Way to be split off. So you can hydrogenolytically e.g. B. with hydrogen in the presence of a noble metal, such as Palladium or platinum catalyst or by acid hydrolysis remove.

The starting materials used are known, or can be produced in a manner known per se. So you can Compounds of formula III z. B by implementing the corresponding in the 3-position of unsubstituted sugar with a halogen R₂-acetic acid, where R₂ has the meaning given above, and obtained their esters in the presence of a strong base. In this halogen is preferably bromine or primarily chlorine.

Another way of making the new glucosamine derivatives is that one in itself known manner, a compound of formula V.

wherein R, R₁ °, R₂, R₄ °, R₆ ° and R₇ ° have the meaning given above have with a compound of the formula  

wherein R₈ ° and R₉ ° have the meaning given above, with the Provided that carboxyl- in the residues R₇ °, R₈ ° and R₉ ° and, if desired, free hydroxy groups by easily removable Protecting groups are protected, condensed and, if necessary splits off existing protecting groups.

The condensation takes place z. B. in such a way that to compound V in the form of the activated carboxylic acid Amino compound VI, or that one with the acid V the compound VI, the amino groups of which are in activated form, implements. The activated carboxyl group can, for example an acid anhydride, preferably a mixed acid anhydride, be an acid amide or an activated ester. As such come especially the above acid anhydrides, Amides or esters in question. The amino group is, for example activated by reaction with a phosphitamide.

The easily removable protective groups also correspond the ones already mentioned. You can in itself are split off in a known manner; e.g. B. hydrogenolytic for example with hydrogen in the presence of a precious metal such as palladium or patine catalyst or by acid hydrolysis.  

The starting materials can be known Way get. So you can z. B. corresponding in 3-position unsubstituted sugar with a halogen R₂-acetamido Implement R₇ ° -acetic acid, or a compound of formula III with an amino-R₇-acetic acid, whose carboxyl group is protected implement in the manner shown above, and split off the protective group.

Another method of implementing the in the 3-position of the sugar residue side chain in being a connection

wherein R, R₁ °, R₄ ° and R₆ ° have the meaning given above, or a derivative thereof with a compound of the formula

where Z is a reactive esterified hydroxy group represents and R₂, R₇ °, R₈ ° and R₉ ° have the meaning given above have, and possibly existing protective groups splits off.  

A reactive esterified hydroxy group is especially one with a strong inorganic or organic Acid esterified hydroxy group, primarily one which is combined with hydrohalic acids, such as chlorine, Bromic or hydroiodic acid is esterified.

The easily removable protective groups correspond the ones already mentioned. You can in known per se Be split off, e.g. B. hydrogenolytically, for example with hydrogen in the presence of a precious metal such as palladium or platinum catalyst or by acid hydrolysis.

The starting materials used in this process variant are known.

You can also get the new connections, if you are in a compound of formula

wherein R, R₂, R₇ °, R₈ ° and R₉ ° have the meaning given above have, and R₅ is an alkylidene or cycloalkylidene group, acidic splits the oxazoline and dioxolane ring and  any existing protective groups are split off and in the optionally released amino group in 2- Position of the sugar molecule introduces the -CO-R residue.

Alkylidene is especially lower alkylidene, such as isopropylidene, and cycloalkylidene primarily cyclopentylidene or cyclohexylidene.

This split also takes place in a manner known per se Way, e.g. B. with an acidic ion exchanger, in particular those with sulfonic acid groups, such as Amberlite® IR-120 (a Styrene resin with strongly acidic sulfo groups) or Dowex® 50 (polystyrene sulfonic acids) or a strong inorganic or organic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid or a sulfonic acid, e.g. B. methanesulfonic acid, or a phenylsulfonic acid optionally substituted in the aromatic ring, such as p-toluenesulfonic acid or trifluoroacetic acid. If you work in the presence of water, you get in 1-position a free hydroxy group; is also one of the carboxyl groups R₈ and / or R₉ with an alcohol, in particular a lower alkanol, esterified, it can, in particular saponify with aqueous acid at a higher temperature.

But it is also possible that with this split the amino group is released in the 2-position of the sugar molecule becomes. In this case the -CO-R residue must be introduced subsequently will. This is done in the usual way by acylation or Sulfonylation.  

Protecting groups can be found in the compounds obtained subsequently on the peptide residue, e.g. B. by hydrogenolysis, such as B. with catalytically excited hydrogen or hydrolysis, split off.

The starting materials used can be z. B. obtained if you put in an appropriate oxazoline, with a free hydroxy group in the 3-position of the sugar residue the R₂- Introduces acetamidopeptide residue in one or more stages.

The compounds obtained can be known per se Be converted into their salts, e.g. B. by Reacting acidic compounds obtained with alkali or alkaline earth metal hydroxides or obtained basic compounds with Acids.

The procedures described above are based on known methods carried out in the absence or preferably in the presence of diluents or solvents, if necessary, under cooling or warming, under increased pressure and / or in an inert gas, such as nitrogen atmosphere.

They are taking into account everyone in the molecule substituents located, if necessary, in particular in the presence of easily hydrolyzable O-acyl residues, especially gentle reaction conditions, such as short reaction times,  Use of mild acidic or basic agents in low concentration, stoichiometric proportions, Choice of suitable catalysts, solvents, temperature and / or printing conditions.

The invention also relates to those embodiments of the process in which a starting material in the form of a reactive derivative or salt used. Such starting materials are preferred from the procedural to the above as special lead valuable connections.

The present invention also relates to pharmaceutical Preparations which contain compounds of formula I. In the pharmaceutical preparations according to the invention are those for enteral, such as oral or rectal, as well as parenteral administration to warm-blooded animals, which the pharmacological agent alone or together with one Contain pharmaceutically applicable carrier material. The dosage of the active ingredient depends on the warm-blooded species, the age and individual condition, as well as the method of application from.  

The new pharmaceutical preparations contain from about 10% to about 95%, preferably from about 20% to about 90% of the active ingredient. Pharmaceutical preparations according to the invention can e.g. B. in unit dosage form, such as drages, tablets, Capsules, suppositories or ampoules are available.

The pharmaceutical preparations of the present Invention are known in a conventional manner, for. B. means conventional mixing, granulating, coating, solution or Lyophilization process produced. Except for the above mentioned types of application can also pharmaceutical Preparations, in particular for oral use, are obtained, by combining the active ingredient with solid carriers, a mixture obtained is optionally granulated, and the mixture or granules, if desired or necessary after addition from suitable auxiliaries, to tablets or drag'e- Cores processed.

Suitable carriers are in particular fillers, like sugar, e.g. B. lactose, sucrose, mannitol or sorbitol, cellulose preparations and / or calcium phosphates, e.g. B. Tricalcium phosphate or calcium hydrogen phosphate, furthermore binders how starch paste using e.g. B. of corn, wheat, Rice or potato starch, gelatin, tragacanth, methyl cellulose,  Hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose and / or polyvinylpyrrolidone, and / or if desired Disintegrants, such as the above strengths, further Carboxymethyl starch, cross-linked polyvinylpyrrolidone, agar, Alginic acid or a salt thereof, such as sodium alginate, aid are primarily flow regulators and lubricants, e.g. B. silica, talc, stearic acid or salts thereof, such as Magnesium or calcium stearate, and / or polyethylene glycol. Drag´e cores are made with suitable, if necessary gastric juice resistant Provide coatings, where u. a. concentrated Sugar solutions, which may contain Arabic gum, Talc, polyvinylpyrrolidone, polyethylene glycol and / or titanium dioxide contain paint solutions in suitable organic solvents or solvent mixtures or, for production of gastric juice-resistant coatings, solutions of suitable ones Cellulose preparations, such as acetyl cellulose phthalate or hydroxypropyl methyl cellulose phthalate, used. The tablets or Drag'e coatings can contain dyes or pigments, e.g. B. for Identification or labeling of different doses of active ingredients, be attached.

The following examples illustrate the above described invention; however, they should include these in their scope do not restrict in any way. Temperatures are in Celsius degrees indicated.  

Example 1

A solution of 4 g of benzyl-2-acetamido-3-O - {[1-L-1-D-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl} - 2-deoxy- α -D-glucopyranoside-benzyl ester in 80 ml of methanol is hydrogenated to a standstill with 0.4 g of 5% palladium on carbon as a catalyst at normal pressure and room temperature. The catalyst is filtered off, washed with a little methanol and the filtrate is evaporated to dryness in a water jet vacuum. The residue is dissolved in 50 ml of distilled water and further hydrogenated to a standstill with 1 g of 5% palladium on carbon as a catalyst at normal pressure and room temperature. The catalyst is filtered off, washed with a little water and the filtrate is evaporated to dryness. The 2-acetamido-3-O - {[1-L-1-D-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl} - 2-deoxy-D-glucose obtained is dried in a high vacuum over phosphorus pentoxide, [ α ] = + 10 ° ± 1 ° (water, c = 0.930).

The starting material can be produced as follows will:

A solution of 9.5 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O-carboxymethyl-2-deoxy- a- D-glucopyranoside in 400 ml of acetonitrile and 3 ml of triethylamine is mixed with 5.3 g N-ethyl-5-phenyl-isoxazolium-3'-sulfonate (Woodwards Reagens K) are added and the mixture is stirred at room temperature until a clear solution is formed. 7.15 g of L-alanyl-D-glutamic acid-1-amide- γ- benzyl ester hydrochloride, 3 ml of triethylamine and 200 ml of acetonitrile are then added and the reaction mixture is stirred for a further 18 hours at room temperature. The crystallized benzyl-2-acetamido-4,6-O-benzylidene-3-O - {[1-L-1-D-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl} - 2-deoxy- α -D -glucopyranoside-benzyl ester is filtered off, washed with semi-saturated sodium bicarbonate solution and water and dried, [ α ] = + 81 ° ± 1 ° (dimethylformamide, c = 0.816).

A solution of 8 g of this compound in 400 ml of 60% acetic acid is kept at 80 ° C for 1 hour. After cooling, the solution is evaporated to dryness, the residue is mixed twice with 50 ml of water and evaporated to dryness. The crystalline residue obtained is stirred with a little water, the crystals are suction filtered and dried. The benzyl-2-acetamido-3-O - {[1-L-1-D-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl} - 2-deoxy- α- D-glucopyranoside-benzyl ester is thus obtained from Mp 200-202 °, [ a ] = + 77 ° ± 1 ° (dimethylformamide, c = 0.599).

Example 2

Analogously to Example 1, the benzyl-2-acetamido-4,6-O-benzylidene-3-O- (1-D-carboxypropyl) -2-deoxy- a - D-glucopyranoside is condensed with L-alanyl-D-glutamic acid. 1-amide γ- benzyl ester hydrochloride and cleaves off the protective groups. The 2-acetamido-3-O- {1-D-1-L-1-D-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylpropyl} -2-deoxy-glucose is thus obtained.

The starting material can be produced as follows:

A solution of 60 g of benzyl-2-acetamido-4,6-O-benzylidene-2-deoxy- α- D-glucopyranoside in 600 ml of dimethylformamide is mixed with 10 g of sodium hydride and 1.5 hours in a nitrogen atmosphere at 45 ° C. touched. After cooling to 0 ° C., 75 ml of D, L- α- bromobutyric acid ethyl ester are added. The reaction mixture is stirred for 1 hour at room temperature and 50 ° C., neutralized with acetic acid and the solvent is evaporated off in a water jet vacuum. The residue is partitioned between methylene chloride and water, the organic phase is washed again with water, dried over magnesium sulfate and the solvent is evaporated off. The residue, which was dried in a high vacuum, is purified by column chromatography on silica gel. Elution with methylene chloride / ethyl acetate (85/15) gives the ethyl benzyl-2-acetamido-4,6-O-benzylidene-3-O-1-D-carboxypropyl) -2-deoxy- α- D-glucopyranoside of [ α ] = + 113 ° ± 1 ° (chloroform, c = 0.5), mp. 154 ° C (from methylene chloride / ether) and Rf value 0.21 and the benzyl-2-acetamido-4,6 -O-benzylidene-3-O- (1-L-carboxypropyl) -2-deoxy- α -D-glucopyranoside ethyl ester of [ α ] = + 42 ° ± 1 ° (chloroform, c = 0.511), mp. 240 ° C (from ethyl acetate) and Rf value 0.04.

A solution of 38.1 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O- (1-D-carboxypropyl) -2-deoxy- α- D-glucopyranoside- ethyl ester in 300 ml of methanol is with 100 ml of 1N sodium hydroxide solution were added and the mixture was kept at 60 ° C. for 1 hour. The solution is then cooled, concentrated to about 150 ml, diluted with 400 ml of ice water and mixed with 100 ml of 1N ice-cold hydrochloric acid. The product which has crystallized out is filtered off with suction, washed with water and dried. The benzyl-2-acetamido-4,6-O-benzylidene-3-O- (1-D-carboxypropyl) -2-deoxy- α- D-glucopyranoside of mp 210-213 ° and [ α ] = + 110 ° ± 1 ° (dimethylformamide, c = 0.554).

The benzyl-2-acetamido-4,6-O-benzylidene-3-O-1-L-carboxypropyl) -2-deoxy- α- D-glucopyranoside of mp 285 ° C. and [ α ] = + 71 ° ± 1 ° (dimethylformamide, c = 0.589).

Example 3

3.77 g 2-phenyl-4,5- [3-O- (1-D-carboxyethyl) -5,6-O-isopropylidene-D-glucofurano] - Δ ²-oxazoline in 60 ml of acetonitrile and 15 ml of dimethylformamide is stirred together with 1.4 ml of triethylamine and 2.55 g of N-ethyl-5-phenyl-isoxazolium-3'-sulfonate for 1.5 hours at 0 ° C, almost everything goes into solution. Then 3.44 g of L-alanyl-D-glutamic acid-1-amide- γ- benzyl ester hydrochloride and a further 1.4 ml of triethylamine are added and the mixture is stirred for 24 hours at room temperature. It is evaporated to the syrup in an oil vacuum and chromatographed on silica gel with a mixture of chloroform and acetone (8/2). A colorless solid syrup is obtained, which crystallizes when triturated with ether. Mp 96-99 ° [ α ] = + 13 ° (in chloroform).

The crystalline benzyl ester is treated with 5% palladium Hydrogenated coal in dioxane at room temperature and normal pressure and delivers the corresponding after evaporation in vacuum Acid as a syrup.

It is stirred in water with 10 ml of Dowex® 50 H⁺ at room temperature for 15 hours. After filtration and freeze-drying, a colorless powder with a decomposition point of 140 ° C. is obtained. The 2-benzoylamino-3-O- {1-D- [1-L- (1-D-carbamoyl- 3-carboxypropyl) -l-carbamoyläthyl] -carbamoyläthyl} - 2-deoxy- α , β- D- Depending on the drying conditions, glucose contains varying amounts of water of crystallization, in the above case after drying at 60 ° C., 1.33 × 10 ^-2 mbar (0.01 Torr), at 15 hours: ½ water.

Example 4

6.0 g of 2-phenyl-4,5- [3-O- (1-D-carboxypropyl) -5,6-O-isopropylidene-D-glucofurano] - Δ ²-oxazoline, 4.08 g of N-ethyl - 5-phenyl-isoxazolium-3'-sulfonate and 2.25 ml of triethylamine are stirred in 100 ml of acetonitrile and 25 ml of dimethylformamide for 1 hour at 0-5 ° C, everything is dissolved. Then 5.55 g of L-alanyl-D-glutamic acid-1-amide- γ- benzyl ester hydrochloride and a further 2.35 ml of triethylamine are added and the mixture is stirred for 48 hours at room temperature. It is evaporated in an oil vacuum and chromatographed on silica gel in a mixture of chloroform and ethanol (19/1). 9 g of the colorless syrup thus obtained are hydrogenated in dioxane with 5% palladium-carbon, filtered off from the catalyst, concentrated in vacuo and hydrolyzed in a mixture of 40 ml of tetrahydrofuran and 30 ml of water with 1.5 ml of trifluoroacetic acid at room temperature. Then the water is evaporated to dryness 4 times in a vacuum, dissolved in water and lyophilized. The 2-benzoylamino-3- O- {1-D- [1-L- (1-D-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylpropyl} -2-deoxy- α , β- D-glucose crystallized with 0.5 mol water, mp 114-152 °, [ α ] = + 17 ° (in methanol).

Example 5

3.63 g 2-phenyl-4,5- [3-O-carboxymethyl-5,6-O-isopropylidene-D-glucofuranol- Δ ²-oxazoline, 3.43 g of L-alanyl-D-glutamic acid 1- amide- γ- benzyl ester hydrochloride, 1.21 g of N-hydroxysuccinimide, 2.16 g of dicyclohexylcarbodiimide and 1.45 ml of triethylamine are dissolved in 40 ml of dimethylformamide and stirred for 24 hours at room temperature. It is evaporated in an oil vacuum, taken up in dichloroethane and water, suctioned off from the precipitated dicyclohexylurea and the organic phase is shaken twice with water and the aqueous phase twice with dichloroethane. After drying and evaporating the organic phases, a solid syrup is obtained which is chromatographed on silica gel in a mixture of chloroform / ethanol (9/1). The peptide ester obtained, which crystallizes on trituration with ether, melts at 167-168 ° C, [ α ] = -5 ° (chloroform).

4.5 g of the ester mentioned are in dioxane 5% palladium-carbon hydrogenated, filtered off from the catalyst and this is extracted with ethanol. The combined filtrates are evaporated and the residue isopropyl alcohol recrystallized. The acid obtained melts 200-207 ° C.

2.85 g of this is stirred in a mixture of 30 ml of water and 15 ml of tetrahydrofuran with 5 ml of Dowex® 50 H⁺ for 15 hours at room temperature, filtered through a hard filter and evaporated to dryness in vacuo. Trituration with ether gives a colorless powder, the 2-benzoylamino-3-O - {[1-L-1-D-carbamoyl-3-carboxypropyl) -1-carbamoylethyl] - carbamoylmethyl} -2-deoxy- α , β -D-glucose, mp 175-177 ° (as hydrate).

In a variation of the one in Acta Chem. Scand. 18, 185 (1964) described method, the starting material manufacture as follows:

100 g 2-phenyl-4,5 [5,6-O-isopropylidene-D-glucofuranol] - Δ ²-oxazoline is dissolved under exclusion of moisture and carbon dioxide in 1 liter of acetonitrile and added in portions 15.2 g of a 55% NaH mineral oil dispersion with good stirring and stirring for an hour at room temperature. Then 42 ml of ethyl chloroacetate are added dropwise at 0 ° C. and another 42 ml after 1.5 hours. After 1.5 hours, another 11.4 g of NaH dispersion are added, the mixture is stirred for half an hour and another 42 drops at 0 ° C. ml of chloroacetate. After a further 2 hours, the mixture is allowed to warm to room temperature and evaporated to a syrup in vacuo - towards the end in an oil vacuum. This is taken up with ether, shaken 3 times with water, the ether phase is dried over sodium sulfate and, after evaporation, 155 g of a brown oil are obtained. This is dissolved in 150 ml of methanol and a solution of 30 g of potassium hydroxide in 150 ml of water is extracted, extracted twice with ether and the ether phase is washed once with water. The water phases are freed from the ether in vacuo and adjusted to pH = 4 with 1N hydrochloric acid on a pH meter.

The precipitating crystalline 2-phenyl-4,5- [3-O- (carboxymethyl) -5,6-O-isopropylidene-D-glucofuranol] - Δ ²- oxazoline is suction filtered, washed with water and dried. 107 g, 93% of theory are obtained. Th., Mp. 186-188 ° C, [ α ] = -9 ° (CHCl₃, c = 0.9), and [ α ] = -23 ° (CHCl₃, c = 3).

Example 6

6.33 g of 2-phenyl-4,5- (3-O-carboxymethyl-5,6-O-isopropylidene-D-glucofurano) - Δ ²-oxazoline, 5.75 g of 2-ethoxy-N-carbäthoxy- 1 , 2-dihydroquinoline (EEDQ) and 9.3 ml of triethylamine are added to a solution of the trifluoroacetate of dibenzyl L-alanyl-D-glutamic acid (obtained from 8.3 g of N-tert-butyloxycarbonyl-L-alanyl-D- dibenzyl glutamic acid with 5.1 ml of trifluoroacetic acid and 2.6 ml of dichloroethane by hydrolysis at 40 ° C. for 4 hours in 70 ml of dichloroethane. The mixture is allowed to react at 40 ° C. for 15 hours, diluted with chloroform, shaken twice with water and the water phases once with chloroform. After drying over sodium sulfate and evaporating the chloroform solution, 19.9 g of an oil are obtained, which is purified over 400 g of silica gel by elution with ether, then with chloroform to acetone 17: 3. This gives pure 2-phenyl-4,5- [3-O- (1-L- {1-D, 3-dibenzyloxycarbonylpropyl} - carbamoylethyl) -carbamoylmethyl-5,6-O-isopropylidene-D-glucofurano] - Δ ²-oxazoline, mp. 113-116 ° C and [ α ] = 47 ° (CHCl₃, c = 1.54).

7 g of the above compound are hydrogenated with 1.8 g 5% Pd / C in a mixture of 80 ml of tetrahydrofuran and 20 ml of water to a standstill, sucks off the catalyst, evaporates in a vacuum and rubs the residue with ether. 4.9 g of dicarboxylic acid are thus obtained as a colorless powder.

4.4 g of the above dicarboxylic acid are stirred for 20 hours at 40 ° C. with 11 ml of ion exchanger Dowex® 50 W × 4 in a mixture of 45 ml of tetrahydrofuran and 20 ml of water. After filtering and clarifying with charcoal (Darco® G 60), the solution is freeze-dried and colorless, amorphous 2-benzamido-2-deoxy-3-O- {1-L- (1-D, 3-dicarboxypropyl) carbamoylethyl] is obtained. - carbamoylmethyl-D-glucopyranose with the optical rotation [ a ] = + 25 ° (H₂O, c = 0.997).

Example 7

Analogously to Example 6, 5.7 g of 2-phenyl-4,5 [3-O-carboxymethyl-5,6-O-isopropylidene-D-glucofurano] - Δ 2 -oxazoline are condensed with 4.9 g of L-seryl-D -glutamic acid- α- amide- γ- tert-butyl ester hydrochloride with the addition of 2.3 ml of triethylamine and 5.2 g of 2-ethoxy-N-carbäthoxy-1,2-dihydroquinoline in 45 ml of dichloroethane. After 18 hours at 40 ° C, crystals formed. Another 50 ml of dichloroethane are added, the mixture is cooled in ice, filtered off with suction and the crystals are washed with cold dichloroethane. The analytically pure, colorless crystals of 2-phenyl-4,5- [3-O- (1-L- {1-D-carbamoyl-3-tert-butyloxycarbonylpropyl} - carbamoyl-2-hydroxyethyl) -carbamoylmethyl-5, 6-O-isopropylidene-D-glucofurano] - Δ ²-oxazoline have mp 187-188 ° C and [α] = + 7 ° (CH₃OH, c = 1.125)..

2 g of this compound are hydrolyzed at room temperature for 20 hours with a mixture of 15 ml of methylene chloride and 5 ml of trifluoroacetic acid. It is evaporated in an oil vacuum, the residue is triturated with ether and the 2-benzamido-2-deoxy-3-O- [1-L- (1-D-carbamoyl-3-carboxypropyl) carbamoyl-2-hydroxyethyl] is obtained. carbamoylmethyl-D-glucopyranose as a beige powder, mp. 100-115 ° C, [ a ] = + 23 ° (H₂O, c = 0.886), which crystallizes with 2 moles of water and 1 mole of trifluoroacetic acid. R _F = 0.28 CHCl₃ to CH₃OH = 1: 1 (thin layer of silica gel).

Example 8 (Reference example)

Analogously to Example 6 condensed 5.25 g of 2-phenyl-4,5- [3-O-carboxymethyl-5,6-O-isopropylidene-D-glucofurano] - Δ ²-oxazoline and the trifluoroacetic acid salt of L-alanyl D-glutamic acid α- n-propylamide γ- benzyl ester - obtained from 6.2 g of N-tert-butyloxy-carbonyl-L-alanyl-D-glutamic acid- α -n-propylamide γ- benzyl ester and 4.2 ml trifluoroacetic acid in 2.5 ml dichloroethane for 6 hours at 40 ° C - in 60 ml dichloroethane with the addition of 7.55 ml triethylamine and 4.8 g 2-ethoxy-N-carbäthoxy-1,2-dihydroquinoline. After 20 hours at 40 ° C., the mixture is diluted with 50 ml of chloroform, shaken twice with water and the water twice with chloroform. After drying and concentrating the chloroform phase, 15 g of an oil are obtained, which is purified over 200 g of silica gel, eluting with ether, then with chloroform to give acetone = 7: 3. 6.4 g of colorless, amorphous substance of R _F 0.35 CHCl₃ to acetone = 7: 3 (thin layer of silica gel) are obtained.

This is hydrogenated to a standstill with 1.8 g of 5% Pd / C in 80 ml of tetrahydrofuran and 20 ml of water, filtered off from the catalyst and concentrated. The acid shows R _F = 0.58 CHCl₃ to CH₃OH = 3: 1 (thin layer of silica gel). Then stirred with 10 ml of Dowex® 50 W × 4 ion exchanger, 50 ml of tetrahydrofuran and 25 ml of water for 15 hours at room temperature and 12 hours at 40 ° C. After filtering, clarifying with charcoal (Darco® G 60), filtering again and freeze-drying, the colorless, amorphous 2-benzamido-2-deoxy-3-O- [1- L- (1-DNn-propyl-carbamoyl-3 is obtained -carboxypropyl) -carbamoyläthyl] - carbamoylmethyl-D-glucopyranose of mp. 65-140 ° C and [ α ] = + 28 ° (water, c = 1.03), R _F = 0.48 CHCl₃ to CH₃OH = 1: 1 (thin layer of silica gel).

Example 9

The 2-benzamido-2-deoxy-3-O- (1-D-carbamoyl-3-carboxypropyl) carbamoylmethyl] -carbamoylmethyl-D-glucose is obtained from 3 g of 2-phenyl-4,5- [3-O - {(1-D-carbamoyl-3-carboxypropyl) - carbamoylmethyl} carbamoylmethyl-5,6-O-isopropylidene-D-glucofurano] - Δ ²-oxazoline by hydrolysis with 1.5 ml of trifluoroacetic acid in a mixture of 15 ml Dimethoxyethane and 15 ml water at 40 ° C for 3 hours. The mixture is concentrated to dryness in vacuo and the residue is extracted again with ether. The remaining powder is dissolved in water, treated with Darco® G 60 charcoal, filtered and freeze-dried. A colorless, amorphous substance of mp 115-155 ° C. and [ α ] = + 34 ° (water, c = 0.81), R _F = 0.28 CHCl₃ to CH₃OH = 1: 1 (thin layer of silica gel) is thus obtained. .

The starting material for this is obtained as follows: 8.0 g of N-tert-butyloxycarbonyl-glycyl-D-glutamic acid- α- amide- γ- benzyl ester is dissolved in a mixture of 6.3 ml of trifluoroacetic acid and 7 ml of dichloroethane and 2 React for days at room temperature and 3 hours at 45 ° C. 12.1 ml of triethylamine, 7.0 g of 2-ethoxy-N-carbäthoxy-1,2-dihydroquinoline (EEDQ) and 8.1 g of 2-phenyl-4,5- [3-O-carboxymethyl -5,6-O-isopropylidene-D-glucofurano] - Δ ²-oxazoline and 20 ml of dimethylformamide. After 20 hours at 40 ° C., the mixture is concentrated in an oil vacuum and the residue is partitioned between methylene chloride and water. After drying and concentrating the methylene chloride phases, a solid residue is obtained, which is extracted twice with ether and recrystallized from toluene. Yield 8.25 g; Mp 157 ° C, [ α ] = + 10 ° (CHCl₃, c = 1.48), R _F = 0.35 (CHCl₃ to ethanol = 9: 1) (thin layer of silica gel).

The benzyl ester thus obtained is hydrogenated to a standstill with 1 g of 5% Pd / C in 100 ml of tetrahydrofuran and 25 ml of water. After filtering off the catalyst and evaporating, the substance is chromatographed on 250 g of silica gel, in CHCl₃: CH₃OH = 4: 1. 5.8 g of colorless, amorphous 2-phenyl-4,5- [3-O - {(1- carbamoyl-3-carboxypropyl) - carbamoylmethyl} -carbamoylmethyl-5,6-O-isopropylidene-D-glucofurano] - Δ ²-oxazoline from R _F = 0.43, CHCl₃ to CH₃OH = 3: 2 (thin layer of silica gel).

Example 10

Analogously to Example 6 condenses 9.5 g of 2-phenyl-4,5- [3-O-carboxymethyl-5,6-O-isopropylidene-D-glucofurano] - Δ ²-oxazoline with 6.25 g of L-alanyl D-glutamic acid α , γ- diamide hydrochloride with the addition of 3.4 ml of triethylamine and 7.95 g of 2-ethoxy-N-carbäthoxy-1,2-dihydroquinoline (EEDQ) in a mixture of 50 ml of dichloroethane and 150 ml Dimethylformamide. The mixture is allowed to react at room temperature for 2 days and at 40 ° C. for 4 hours. The mixture is concentrated under an oil vacuum and the residue is extracted first twice with ether, then twice with ice water. After drying, the product can be recrystallized from dichloroethane. This gives colorless crystals of mp. 170-184 ° C; [ α ] = + 3 ° (DMSO, c = 1.43) R _F = 0.64 CHCl₃ to CH₃OH = 3: 1 (thin layer of silica gel).

6.1 g of this compound are hydrolyzed with 13.5 ml of Dowex® 50 ion exchanger in a mixture of 60 ml of dimethoxyethane and 60 ml of water for 15 hours at room temperature. After filtering and concentrating, the residue is taken up in water, clarified with Darco® G 60 charcoal, suction filtered and the filtrate freeze-dried. Colorless, amorphous 2-benzamido-2-deoxy-3-O- [1-L-1-D-dicarbamoylpropyl) - carbamoylethyl] -carbamoylmethyl-D-glucopyranose of mp 82-143 ° C .; [ α ] = + 24 ° (H₂O, c = 0.98), R _F = 0.45 CHCl₃ to CH₃OH = 1: 1 (thin layer of silica gel). The substance crystallizes with 1.23 mol water of crystallization.

Example 11

Analogously to Example 10, 2-benzamido-2-deoxy-3-O- [1-L-1-D-bis-N-methylcarbamoylpropyl is obtained from 2-benzamido-2-deoxy-3-O-carboxymethyl-D-glucopyranose ) - carbamoylethyl] -carbamoylmethyl-D-glucopyranose of mp 125-132 ° C and [ α ] = + 24 ° (H₂O, c = 0.93) R _F = 0.26 to ethanol = 7: 3 (silica gel thin-layer plates) .

Example 12

Analogously to Example 10, 2-benzamido-2-deoxy-3-O- [2-benzamido-2-deoxy-3-O-carboxymethyl-D-glucopyranose and the hydrochloride of L-alanyl-D-glutamic acid dimethyl ester are obtained. 1-L-1-D-bis-methoxycarbonylpropyl) - carbamoylethyl] -carbamoylmethyl-D-glucopyranose as hydrate, mp 80-90 ° C, [ α ] = + 25 ° (CH₃OH, c = 1.017), R _F - Value = 0.23 in CHCl₃ to ethanol = 9: 1 (silica gel thin-layer plates).

Example 13

Analogously to Example 8, the hydrochloride of L- α- amino-valeroyl-D-glutamic acid- α- amide γ- tert-butyl ester gives the 2-benzamido-2-deoxy-3-O- [1-L- ( 1-D-carbamoyl-3-carboxypropyl) -carbamoylbutyl] -carbamoylmethyl-D-glucopyranose.

Example 14

Analogously to Example 8, the 2-benzamido-2- deoxy-3-O- [1-L-1-D-carbamoyl-3-carboxypropyl) carbamoylpropyl] - carbamoylmethyl-D-glucopyranose, m.p. 114-155 ° C.

Example 15

Analogously to Example 8, 2-benzamido-2-deoxy-3-O- [1-L-1-D-carbamoyl-3-carboxypropyl) -carbamoyl-2-methylpropyl] -1-carbamoylmethyl-D-glucopyranose, [ α ] = = 32 ° ( c = 0.78; water).

Example 16

A solution of 10.7 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O- [1-D-carbamoyl-3-carboxypropyl) -carbamoylmethyl-carbamoylmethyl] -2-deoxy- α -D- Glucopyranoside benzyl ester in 200 ml glacial acetic acid and 100 ml water is hydrogenated with 2 g 5% palladium on carbon at normal pressure and room temperature for 57 hours. The catalyst is filtered off, washed with water and the filtrate is evaporated. The residue is taken up in water, filtered through 100 ml of ion exchanger Amberlite® IR 120 (H⁺ form) and the filtrate freeze-dried. The 2-acetamido-3-O - [(1-D-carbamoyl-3-carboxypropyl) -carbamoylmethyl-carbamoylmethyl] - 2-deoxy-D-glucose is crystallized from methanol / ethyl acetate and dried under high vacuum. The product, which contains ¼ mol of ethyl acetate, shows the optical rotation [ α ] = + 27 ° ± 1 ° (water, c = 0.944).

The starting material used can be as follows getting produced:

8 g of Nt-butyloxycarbonyl-glycyl-D-isoglutamine benzyl ester is dissolved at room temperature and with the exclusion of moisture in a mixture of 18 ml of 1,2-dichloroethane and 8.4 ml of trifluoroacetic acid and left to stand for 16 hours. The reaction mixture is then diluted with 200 ml of tetrahydrofuran, neutralized with triethylamine with external cooling and with a solution of 8.3 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O-carboxymethyl-2-deoxy- α - D-glucopyranoside in 100 ml of tetrahydrofuran and 2.52 ml of triethylamine. After the addition of 5.05 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ), the mixture is stirred for 24 hours at room temperature. The product which has crystallized out is filtered off with suction, washed with tetrahydrofuran and ether and dried. The benzyl-2-acetamido-4,6-O-benzylidene-3-O- [1-D-carbamoyl-3-carboxypropyl) -carbamoylmethyl-carbamoylmethyl] -2-deoxy- a- D-glucopyranoside-benzyl ester obtained has this optical rotation [ α ] = + 66 ° ± 1 ° (N, N-dimethylformamide, c = 1.308).

Example 17

A solution of 6.8 g of benzyl-2-acetamido-3-O - {[1-L- (1-D-1,3-dicarbamoylpropyl) carbamoylethyl] carbamoylmethyl} - 2-deoxy- α- D-glucopyranoside in 200 ml of 50% aqueous methanol is hydrogenated with 5% palladium on carbon at normal pressure and room temperature for 60 hours. The catalyst is filtered off and the filtrate is evaporated. The residue is taken up in 50 ml of water and lyophilized. This gives 2-acetamido-3-O - {[1-L-1-D- (1,3-dicarbamoylpropyl) carbamoylethyl] carbamoylmethyl} -2-deoxy-D-glucose, which is 1.24 mol of water contains, as a white powder of [ α ] = + 7 ° ± 1 ° (water, c = 0.514).

The starting material can be produced as follows will:

A solution of 9.1 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O-carboxymethyl-2-deoxy- α- D-glucopyranoside in 100 ml of N, N-dimethylformamide and 2.77 ml of triethylamine 5.0 g of L-alanyl-D-glutamic acid diamide hydrochloride and 5.1 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) are added and the mixture is left to stand at room temperature for 48 hours. After the solvent has been distilled off, the residue is extracted thoroughly with ether and water, dried and recrystallized from chloroform / methanol, [ α ] = + 83 ° ± 1 ° (N, N-dimethylformamide, c = 0.531).

A solution of 4 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O - {[1-L-1,3-D-dicarbamoylpropyl) carbamoylethyl] - carbamoylmethyl} -2-deoxy- α - D-glucopyranoside in 120 ml glacial acetic acid is diluted with 80 ml water and stirred at 60 ° C for 3 hours. The reaction mixture is then cooled, evaporated, 100 ml of water are added to the residue three times and this is distilled off in each case. The benzyl-2-acetamido-3-O - {[1-L- (1,3-D-dicarbamoylpropyl) carbamoylethyl] -carbamoylmethyl} -2-deoxy- a- D-glucopyranoside obtained is recrystallized from methanol, mp. 223-225 ° C.

Example 18

A solution of 5.7 g benzyl-2-acetamido-3-O- {1-D- [(1-D-carbamoyl-3-carboxypropyl) carbamoylmethyl] carbamoylpropyl} - 2-deoxy- α- D-glucopyranoside -benzyl ester in 100 ml glacial acetic acid is hydrogenated in the presence of 5% palladium on carbon at normal pressure and room temperature. The catalyst is filtered off and the filtrate is evaporated. The remaining 2-acetamido-3-O- {1-D [(1-D-carbamoyl-3-carboxypropyl) carbamoylmethyl] -carbamoylpropyl} -2-deoxy-D-glucose is taken up in 50 ml of water and freeze-dried, [ α ] = + 46 ° ± 1 ° (water, c = 0.630).

The starting material used can be as follows getting produced:

5.1 g of Nt-butyloxycarbonyl-glycyl-D-isoglutamine-benzyl ester are dissolved at room temperature and with exclusion of moisture in a mixture of 5.1 ml of 1,2-dichloroethane and 5.1 ml of trifluoroacetic acid and left to stand for 16 hours. This solution is diluted with 100 ml of tetrahydrofuran, neutralized with external cooling with triethylamine and a solution of 6.3 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O- (1-D-carboxypropyl) -2 -deoxy- α -D-glucopyranoside and 1.8 ml of triethylamine in 100 ml of tetrahydrofuran and 3.2 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ). After 24 hours, the mixture is evaporated to dryness and the residue is partitioned between ethyl acetate and water. The organic phase is washed with ice-cold 2N hydrochloric acid, water, a saturated sodium hydrogen carbonate solution and water and evaporated. The remaining benzyl-2-acetamido-4,6-O-benzylidene-3-O- {1-D-1-D-carbamoyl-3-carboxypropyl) -carbamoylmethyl] - carbamoylpropyl} -2-deoxy- α -D- Glucopyranoside benzyl ester is crystallized from tetrahydrofuran / ether, [ α ] = + 76 ° ± 1 ° (N, N-dimethylformamide, c = 0.457).

A mild acid hydrolysis of this product in 60% aqueous acetic acid leads to benzyl-2-acetamido-3-O {1-D- [(1-D-carbamoyl-3-carboxypropyl) -carbamoylmethyl] -carbamoylpropyl} - 2-deoxy- α- D-glucopyranoside-benzyl ester, which crystallizes from methanol / ether, mp. 180-185 ° C. [ α ] = + 87 ° ± 1 ° (methanol, c = 0.457).

Example 19

4 g of Nt-butyloxycarbonyl-L-alanyl-D-isoglutamine-benzyl ester is dissolved at room temperature and with exclusion of moisture in a mixture of 4 ml of trifluoroacetic acid and 4 ml of 1,2-dichloroethane and left to stand for 16 hours. This reaction mixture is now diluted with 30 ml of 1,2-dichloroethane, neutralized with external cooling with triethylamine and with a solution of 3.7 g of benzyl-2-acetamido-3-O-carboxymethyl-2-deoxy- α- D-glucopyranoside and 1.38 ml of triethylamine in 100 ml of tetrahydrofuran. After the addition of 2.6 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ), the mixture is left to stand at room temperature for 24 hours and evaporated to dryness. The residue is dissolved in chloroform / methanol 9/1 and this solution is washed with water, ice-cold 2N hydrochloric acid, water, a saturated sodium hydrogen carbonate solution and water and the solvent is evaporated off. The benzyl-2-acetamido-3-O - {[1-L- (1-D-carbamoyl-3-carboxypropyl) - carbamoylethyl] -carbamoylmethyl} -2-deoxy- α- D-glucopyranoside benzyl ester thus obtained is converted from Recrystallized ethanol, mp. 208-212 ° C, [ α ] = + 77 ° ± 1 ° (N, N-dimethylformamide, c = 0.546).

After the two benzyl radicals have been split off hydrogenolytically, as described in Example 1, the 2-acetamido-3-O - {[1-L- (1-D-carbamoyl-3-carboxypropyl) -carbamoylethyl] - carbamoylmethyl} -2 is obtained -deoxy-D-glucose, [ α ] = + 10 ° (water: c = 0.892).

Example 20

A solution of 3.8 g of benzyl-2-acetamido-3-O - {[1-L- (1-D-1,3-dicarboxypropyl) carbamoylethyl] carbamoylmethyl} -2-deoxy- α- D-glucopyranoside -dimethyl ester in 100 ml of methanol is hydrogenated in the presence of 5% palladium on carbon at normal pressure and room temperature. When the uptake of hydrogen has ended, the catalyst is filtered off and the filtrate is evaporated to dryness. The residue is taken up in 70 ml of water and freeze-dried. The foam obtained is the 2-acetamido-3-O - {[1-L- (1,3-D-dicarboxypropyl) carbamoylethyl] carbamoylmethyl} -2-deoxy-D-glucose dimethyl ester with [ α ] = + 23 ° ± 1 ° (water, c = 0.814).

The starting material used can be produced as follows will:

A solution of 12.9 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O-carboxymethyl-2-deoxy- α- D-glucopyranoside and 4.0 ml of triethylamine in 100 ml of N, N-dimethylformamide 8.1 g of dimethyl L-alanyl-D-glutamic acid hydrochloride and 6.95 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) are added and the mixture is left to stand for 20 hours at room temperature. The solvent is then evaporated off, the residue is taken up in chloroform and this solution is washed with water, ice-cold 2N hydrochloric acid, water, a saturated sodium hydrogen carbonate solution and water. After drying over magnesium sulfate, the mixture is evaporated to dryness. The residue is extracted with warm ethanol, the undissolved benzyl-2-acetamido-4,6-O-benzylidene-3-O - {[1-L-1,3-D-dicarboxypropyl) - carbamoylethyl] -carbamoylmethyl} -2 -deoxy- α -D- glucopyranoside dimethyl ester filtered off and dried, [ α ] = + 22 ° ± 1 ° (chloroform, c = 1.160).

A solution of 15.6 g benzyl-2-acetamido-4,6-O-benzylidene-3-O - {[1-L- (1,3-D-dicarboxypropyl) carbamoylethyl] carbamoylmethyl} -2-deoxy - α -D-glucopyranoside dimethyl ester in 420 ml of glacial acetic acid and 280 ml of water is heated to 10 ° C. After 4 hours of stirring at this temperature, the reaction mixture is cooled and evaporated to dryness. The residue is mixed three times with 100 ml of water and evaporated to dryness. The residue is taken up in chloroform, this solution washed with water, dried over magnesium sulfate and evaporated to dryness. The benzyl-2-acetamido-3-O - {[1-L- (1,3-D-dicarboxypropyl) -carbamoylethyl] -carbamoylmethyl} -2-deoxy- α -D-glucopyranoside dimethyl ester is thus obtained as a yellowish resin from [ α ] = + 31 ° ± 1 ° (chloroform, c = 1.070).

Example 21

A solution of 6.1 g benzyl-3-O - {[1-L- (1-D-carbamoyl-3-carboxypropyl) carbamoylethyl] carbamoylmethyl} -2-deoxy-2-propionamido- α- D-glucopyranoside -benzyl ester in 200 ml of tetrahydrofuran: water 2: 1 is hydrogenated in the presence of 0.6 g of 5% palladium on carbon at normal pressure and room temperature. When the uptake of hydrogen has ended, the catalyst is suctioned off and the filtrate is evaporated. The residue is mixed with 150 ml of water and hydrogenated in the presence of 5% palladium on carbon until no more hydrogen is taken up. The catalyst is filtered off and lyophilized. The 3-O - {[1-L- (ID-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl} 2-deoxy-2-propionamido-D-glucose of [ α ] = + 8 ° (water ; c = 1.146).

The starting material used can be as follows getting produced:

A solution of 90 g of benzyl-2-acetamido-2-deoxy- α- D-glucopyranoside in 900 ml of N, N-dimethylformamide is mixed with 0.3 ml of methanesulfonic acid with stirring, moisture exclusion and ice-water cooling. A solution of 60 ml of isopropenylmethyl ether in 240 ml of N, N-dimethylformamide is then added dropwise over an hour, the mixture is stirred at room temperature for two hours and made alkaline with triethylamine. The solvent is distilled off, the residue is taken up in ethyl acetate, this solution is washed with water, dried over magnesium sulfate and evaporated to dryness. The product, the benzyl-2-acetamido-2-deoxy-4,6-O-isopropylidene- α- D-glucopyranoside is crystallized from ether, mp. 136-137 ° C, [ α ] = + 103 ° ± 1 ° (Chloroform, c = 1.125).

52.5 g of benzyl-2-acetamido-2-deoxy-4,6-O-isopropylidene- α- D-glucopyranoside are dissolved in a solution of 225 g of potassium hydroxide in 750 g of ethanol and 40 ml of distilled water and refluxed for 4½ hours heated. After cooling, the reaction mixture is concentrated to half and poured onto ice. It is extracted with chloroform, the organic phase is washed with water, dried over magnesium sulfate and evaporated to dryness. The residue of the benzyl-2-amino-2-deoxy-4,6-O-isopropylidene- α- D-glucopyranoside is crystallized from ether, mp. 145-146 ° C, [ α ] = + 117 ° ± 1 ° ( Chloroform, c = 1.295).

A solution of 24.7 g of benzyl-2-amino-2-deoxy-4,6-O-isopropylidene- α- D-glucopyranoside in 192 ml of chloroform is mixed with a solution of 16.0 g of potassium hydrogen carbonate in 192 ml of distilled water and cooled to 0 ° C. 8.16 g of propionic acid chloride are then added dropwise over the course of 20 minutes, and the mixture is stirred at this temperature for a further 50 minutes. Now the organic phase is separated off, washed with water, dried over magnesium sulfate and evaporated to dryness. The product, the benzyl-2-deoxy-4,6-O-isopropylidene-2-propionamido- α- D-glucopyranoside, is crystallized from ethyl acetate / petroleum ether, mp. 121-122 ° C, [ α ] = + 112 ° ± 1 ° (chloroform, c = 0.977).

A solution of 9.1 g of benzyl-2-deoxy-4,6-O-isopropylidene-2-propionamido- α- D-glucopyranoside in 90 ml of acetonitrile is practically mixed with 1.25 g of sodium hydride. added and stirred at 40 ° C for 2 hours. Then it is cooled to -5 to -10 ° C and mixed with 4.2 ml of bromoacetic ester. After a further 20 minutes, 10 ml of ethanol are added, the reaction mixture is neutralized with glacial acetic acid and evaporated to dryness. The residue is partitioned between ether and water, the ether solution washed with water, dried over magnesium sulfate and evaporated. The product, the benzyl-3-O-carboxymethyl-2-deoxy-4,6-O-isopropylidene-2-propionamido-- α- D-glucopyranoside ethyl ester is crystallized from ether / petol ether, mp. 94-95 ° C , [ α ] = + 145 ° ± 1 ° (chloroform, c = 1.218).

A solution of 6.8 g of benzyl-3-O-carboxymethyl-2-deoxy-4,6-O-isopropylidene-2-propionamido- α- D-glucopyranoside ethyl ester in 70 ml of methanol is mixed with 22.5 ml of 1N Sodium hydroxide solution added. When the ester hydrolysis has ended, 7.5 ml of 1N hydrochloric acid are added and the mixture is evaporated to dryness. The product is dissolved in 50 ml of N, N-dimethylformamide and condensed with 15 mmol of L-alanine-D-isoglutaminobenzyl ester trifluoroacetate in the presence of 3.72 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ). The mixture is then evaporated to dryness and the residue is taken up in chloroform. This solution is washed with water, ice-cold 2N hydrochloric acid, water, a saturated sodium hydrogen carbonate solution and water, dried over magnesium sulfate and evaporated to dryness. The product is crystallized from dilute ethanol, mp. 177-80 ° C, [ α ] = + 71 ° ± 1 ° (chloroform, c = 1.047).

A solution of 8.1 g of benzyl-3-O - {[1-L-1-D-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl} -2-deoxy-4,6-O-isopropylidene-2- 15 ml of 1N hydrochloric acid are added to propionamido- α- D-glucopyranoside benzyl ester in 150 ml of methanol and the mixture is left to stand for one hour at room temperature. 15 ml of 1N sodium hydroxide solution are now added and the mixture is evaporated to dryness. The benzyl-3-O - {[1-L-1-D-carbamoyl-3-carboxypropyl) - carbamoylethyl] -carbamoylmethyl} -2-deoxy-2-propionamido- α- D-glucopyranoside-benzyl ester obtained is obtained from methanol / Water crystallized and dried, mp. 208-210 ° C, [ α ] = + 75 ° ± 1 ° (N, N-dimethylformamide, c = 1.120).

Example 22

A 5% solution of benzyl-2-acetamido-2-deoxy-3-O - {[1-L-1,3-D-bis-methylcarbamoylpropyl) carbamoylethyl] - carbamoylmethyl} - α -D-glucopyranoside in distilled Water / methanol 1: 1 is hydrogenated with 5% palladium on carbon, the catalyst is filtered off and the filtrate is evaporated. The product, the 2-acetamido-2-deoxy-3-O - {[1-L-1,3-D-bis-methyl-carbamoylpropyl) -carbamoylethyl] -carbamoylmethyl} -D-glucose is freeze-dried, [ α ] = + 31 ° (water: c = 0.41).

The starting material can be produced as follows will:

A solution of 9.1 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O-carboxymethyl-2-deoxy- α- D-glucopyranoside in 100 ml of N, N-dimethylformamide and 2.77 ml of triethylamine 5.6 g of L-alanyl-D-glutamic acid bis-methylamide hydrochloride and 5.1 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline are added and the mixture is left to stand at room temperature for 48 hours. The solvent is then distilled off, water is added to the oily residue, the insolubles are filtered off, washed with water and dried. This product is stirred twice more with ether, filtered off and dried and provides the benzyl-2-acetamido-4,6-O-benzylidene-2-deoxy-3-O - {[1-L-1,3-D-bis -methylcarbamoylpropyl) -carbamoyläthyl] -carbamoylmethyl} - α -D-glucopyranoside.

By mild acid hydrolysis, as described in Example 17, with 60% acetic acid, the benzylidene group is split off and the product, the benzyl-2-acetamido-2-deoxy-3-O - {[1-L-1,3-D -bis-methylcarbamoylpropyl) -carbamoyläthyl] - carbamoylmethyl} - α -D-glucopyranoside isolated.

Example 23

A 5% aqueous solution of benzyl-2-acetamido-3-O - {[1-L-1-D-carbamoyl-3-carboxypropyl) -carbamoyl-2-hydroxyethyl] - carbamoylmethyl} -2-deoxy- α - D-glucopyranoside is hydrogenated in the presence of 5% palladium on carbon, filtered and freeze-dried. The 2-acetamido-3-O - {[1- L-1-D-carbamoyl-3-carboxypropyl) -carbamoyl-2-hydroxyethyl] -carbamoylmethyl} -2-deoxy-D-glucose of [ α ] = + 10 ° (water: c = 1.653).

The starting material can be produced as follows will:

A solution of 3.7 g of benzyl-2-acetamido-3-O-carboxymethyl-2-deoxy- α- D-glucopyranoside and 1.38 ml of triethylamine in 150 ml of tetrahydrofuran is mixed with 3.26 g of L-serine-D- Isoglutamine tert-butyl ester hydrochloride and 2.6 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) were added and the mixture was left to stand at room temperature for 20 hours. After evaporation of the solvent, the residue is dissolved in chloroform: methanol 9: 1, washed with water, ice-cold 2N hydrochloric acid, water, a saturated sodium bicarbonate solution and water, filtered and freed from the solvent. The benzyl-2-acetamido-3-O - {[1-L-1-D-carbamoyl-3-carboxypropyl) - carbamoyl-2-hydroxyethyl] -carbamoylmethyl} -2-deoxy- a- D-glucopyranoside is thus obtained - tert-butyl ester.

By mild acidic hydrolysis as described in Example 7, the tert-butyl ester is hydrolyzed and the benzyl-2-acetamido-3-O - {[1-L-1-D-carbamoyl-3-carboxypropyl) carbamoyl-2 -hydroxyethyl] -carbamoylmethyl} -2-deoxy- α -D-glucopyranoside obtained.

Example 24

A 5% solution of benzyl-2-acetamido-3-O - {[1- L-1-D-carbamoyl-3-carboxypropyl) carbamoylbutyl] carbamoylmethyl} - 2-deoxy- α- D-glucopyranoside in methanol : Water 1: 1 is hydrogenated in the presence of 5% palladium on carbon, filtered and evaporated. The residue is dissolved in distilled water and freeze-dried. The 2-acetamido-3-O - {[1-L-1-D-carbamoyl-3-carboxypropyl) -carbamoylbutyl] -carbamoylmethyl} -2-deoxy-D-glucose, [ α ] (subsequently submitted) = + 12 ° (water, c = 0.722).

The starting material can be produced as follows will:  

A solution of 3.7 g of benzyl-2-acetamido-3-O-carboxymethyl-2-deoxy- α- D-glucopyranoside and 1.38 ml of triethylamine in 100 ml of tetrahydrofuran is mixed with 3.28 g of L-norvaline-D- Isoglutamine tert-butyl ester hydrochloride and 2.6 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) were added and the mixture was left to stand at room temperature for 24 hours. The reaction mixture is evaporated to dryness and the residue is dissolved in chloroform. The solution obtained is washed with water, ice-cold 2N hydrochloric acid, water, a saturated sodium hydrogen carbonate solution and water, dried over magnesium sulfate and evaporated. The benzyl-2-acetamido-3-O - {[1-L- (1-D-carbamoyl-3-carboxypropyl) - carbamoylbutyl] -carbamoylmethyl} -2-deoxy- α -D-glucopyranoside- tert.- butyl ester, m.p. (subsequently) 202-204 ° C.

The tert-butyl ester is by mild acid hydrolysis split.

Example 25

A 5% aqueous solution of benzyl-2-acetamido-3-O - {[1-L- (1-D-carbamoyl-3-carboxypropyl) carbamoyl-2-methylpropyl] carbamoylmethyl} -2-deoxy- α -D-glucopyranoside is hydrogenated in the presence of 5% palladium on carbon, filtered and the filtrate lyophilized. This gives the 2-acetamido-3-O - {[1-L- (1-D-carbamoyl-3-carboxypropyl) carbamoyl-2-methylpropyl] carbamoylmethyl-2-deoxy-D-glucose, [ α ] (submitted later) = + 11 ° (water, c = 1,096).

The starting material can be produced as follows will:

A solution of 3.7 g of benzyl-2-acetamido-3-O-carboxymethyl-2-deoxy- α- D-glucopyranoside and 1.38 ml of triethylamine in 100 ml of tetrahydrofuran is mixed with 3.38 g of L-valine-D- Isoglutamine tert-butyl ester hydrochloride and 2.6 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline were added, left to stand at room temperature for 24 hours and worked up as described in Example 24. This gives benzyl-2-acetamido-3-O - {[1-L-1-D-carbamoyl-3-carboxypropyl) -carbamoyl-2-methylpropyl] -carbamoylmethyl} -2-deoxy- a -D-glucopyranoside- tert-butyl ester, which is mildly acidic to benzyl-2-acetamido-3-O - {[1-L-1-D-carbamoyl-3-carboxypropyl) -carbamoyl-2-methylpropyl] -carbamoylmethyl} -2-deoxy- α -D-glucopyranoside is hydrolyzed.

Example 26

A 5% solution of benzyl-2-acetamido-3-O - {[1- L- (1-D-carbamoyl-3-carboxypropyl) -carbamoyl-3-methylbutyl] - carbamoylmethyl} -2-deoxy- α - D-glucopyranoside in 50% aqueous methanol is hydrogenated in the presence of 5% palladium on carbon, filtered and evaporated. The residue is dissolved in distilled water, filtered again and freeze-dried. The 2-acetamido-3-O- {[1-L- (1-D-carbamoyl-3-carboxypropyl) -carbamoyl-3-methylbutyl] carbamoylmethyl} -2-deoxy-D-glucose, [ α ] (submitted later) = + 16 ° (water, c = 1.06).

The starting material used can be as follows getting produced:  

A solution of 3.7 g of benzyl-2-acetamido-3-O-carboxymethyl-2-deoxy- α- D-glucopyranoside and 1.58 ml of triethylamine in 100 ml of tetrahydrofuran is mixed with 3.52 g of L-leucine-D- Isoglutamine tert-butyl ester hydrochloride and 2.6 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) were added, left to stand at room temperature for 24 hours and worked up as described in Example 24. The benzyl-2-acetamido-3-O - {[1-L- (1-D-carbamoyl-3-carboxypropyl) -carbamoyl-3-methylbutyl] -carbamoylmethyl} -2-deoxy- α- D-glucopyranoside is obtained - tert-butyl ester, its mild acid hydrolysis to benzyl-2-acetamido-3-O - {[1-L- (1-D-carbamoyl-3-carboxypropyl) - carbamoyl-3-methylbutyl] -carbamoylmethyl} -2 -deoxy- α -D-glucopyranoside leads.

Example 27

A 5% solution of benzyl-2-acetamido-3-O - {[1- L- (1-D-carbamoyl-3-carboxypropyl) carbamoylpropyl] carbamoylmethyl} - 2-deoxy- α- D-glucopyranoside in 50% aqueous methanol is hydrogenated in the presence of 5% palladium on carbon at normal pressure and room temperature, filtered off from the catalyst and evaporated. The residue is dissolved in distilled water and freeze-dried. The 2-acetamido-3-O - {[1-L- (1-D-carbamoyl-3-carboxypropyl) -carbamoylpropyl] - carbamoylmethyl} -2-deoxy-D-glucose, [ α ] (subsequently submitted) is thus obtained. = + 11 ° (water, c = 1.019).

The starting material can be produced as follows will:

A solution of 3.7 g of benzyl-2-acetamido-3-O-carboxymethyl-2-deoxy- α- D-glucopyranoside and 1.38 ml of triethylamine in 100 ml of tetrahydrofuran is mixed with 3.24 g of N- α -L- Aminobutyryl-D-isoglutamine tert-butyl ester hydrochloride and 2.6 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline were added and worked up after 24 hours at room temperature, as described in Example 23. The resulting benzyl-2-acetamido-3-O - {[1-L- (1-D-carbamoyl-3-carboxypropyl) -carbamoylpropyl] -carbamoylmethy-l} - 2-deoxy- α -D-glucopyranoside-tert. -butyl ester is then mildly acidic to benzyl-2-acetamido-3-O - {[1-L- (1- D-carbamoyl-3-carboxypropyl) -carbamoylpropyl] -carbamoylmethyl} - 2-deoxy- α -D-glucopyranoside hydrolyzed.

Example 28

A 5% solution of benzyl-2-acetamido-3-O - {[1-L- (1-D-carbamoyl-3-carboxypropyl) -carbamoyl-phenylmethyl] -carbamoylmet-hyl} - 2-deoxy- α - D-glucopyranoside in 50% aqueous methanol is hydrogenated in the presence of 5% palladium on carbon at normal pressure and room temperature, filtered off from the catalyst and evaporated. The residue is dissolved in distilled water and freeze-dried and the 2-acetamido-3-O - {[1-L- (1-D-carbamoyl-3-carboxypropyl) -carbamoyl-phenylmethyl] -carbamoylmethyl} -2-deoxy- D-glucose.

The starting material can be produced as follows will:

A solution of 3.7 g of benzyl-2-acetamido-3-O-carboxymethyl-2-deoxy- α- D-glucopyranoside and 1.38 ml of triethylamine in 100 ml of tetrahydrofuran is mixed with 3.72 g of l-phenylglycine-D- Isoglutamine tert-butyl ester hydrochloride and 2.6 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) were added, left to stand at room temperature for 24 hours and worked up as described in Example 23. The benzyl-2-acetamido-3-O - {[1-L- (1-D-carbamoyl-3-carboxypropyl) - carbamoyl-phenylmethyl] -carbamoylmethyl} -2-deoxy- α- D-glucopyranoside is thus obtained. tert-butyl ester, which is mildly acidic to benzyl-2-acetamido-3-O - {[1-L- (1-D-carbamoyl-3-carboxypropyl) -carbamoyl-phenylmethyl] -carbamoylmethyl} -2-deoxy- α -D-glucopyranoside is hydrolyzed.

Example 29

3.2 g of 2-amino-2-deoxy-3-carboxymethyl-D-glucopyranose, 2.5 g of K₂CO₃ and 4.5 g of pivalic acid β- nitrophenyl ester are stirred in 100 ml of dimethylformamide at 100 ° C for 15 hours. The salts are then filtered off, the filtrate is evaporated in an oil vacuum and, in aqueous solution, passed through Amberlite® IR-120 ion exchanger. The eluate is shaken out with CHCl₃ and evaporated to a syrup in vacuo. Trituration with ethyl acetate produces a pale yellow powder of 2-pivaloylamido-2-deoxy-3-carboxymethyl-D-glucopyranose, mp = 89-93 ° C.

The starting substance, 2-amino-2-deoxy-3-carboxymethyl-D-glucopyranose, is obtained as follows: 100 g of 2-phenyl-4,5- (3-O-carboxymethyl-5,6-isopropylidene-D-glucofurano ) - Δ ²-oxazoline in 1000 ml of 2.6 N hydrochloric acid is refluxed for 4 hours. The precipitated benzoic acid is filtered off and the filtrate is extracted 3 times with ether. The water phase is adjusted on the pH meter with 3 N KOH to pH = 7 and clarified with activated carbon. The mixture is then concentrated in vacuo to about 500 ml and mixed with 3 l of acetone, cooled with ice and suctioned off from the precipitate. After drying at 40 ° C. in vacuo, 247 g of a yellowish powder are obtained, which consists of 85% KCl and 15% title compound.

22 g of a mixture are obtained from the mother liquor from the title compound and the associated lactam.

The reaction of 2-pivaloylamido-2-deoxy-3-carboxymethyl-D-glucopyranose with L-alanyl-D-glutamic acid- α- amide- γ -benzyl ester-trifluoroacetate is carried out analogously to Example 21 with 2-ethoxy-N-carbäthoxy-1 , 2-dihydroquinoline in DMF: CH₂Cl₂ = 1: 1. After hydrogenation with 5% Pd-C, 2-trimethylacetamido-2-deoxy-3-O - {[1-L- (1-D-carbamoyl-3 -carboxypropyl) -1-carbamoylethyl] - carbamoylmethyl} - α , β -D-glucose as lyophilisate. [ α ] (submitted later) = -7 ° ( c = 0.6; methanol).

Claims (5)

1. Glucosamine derivatives of the general formula wherein RC _1-4 alkyl or phenyl, R₂ hydrogen or C _1-4 alkyl, R₇ hydrogen, C _1-4 alkyl, hydroxymethyl, mercaptomethyl or phenyl and R₈ and R₉ independently carboxyl, (C _1-4 alkoxy ) -carbonyl, carbamoyl or N-methyl-carbamoyl, with the proviso that the alkyl radical R has more than 1 carbon atom if the radical R₂ is methyl, or if the radical R₂ is hydrogen and R₈ and R₉ each represent a carboxyl group, and their salts. 2. 2-acetamido-3-O - {[l-L-l-D-carbamoyl-3-carboxypropyl) - carbamoylethyl] -carbamoylmethyl} -2-deoxy-D-glucose or a pharmaceutically acceptable salt thereof according to claim 1. 3. A process for the preparation of glucosamine derivatives and their salts according to claim 1, characterized in that in a manner known per se

• a) a compound of the formula wherein R has the abovementioned meaning and R₁ °, R₄ ° and R₆ ° represent an easily removable protective group, or a derivative thereof, with a compound of the formula VIII wherein Z represents a reactive esterified hydroxyl group, R₂ has the meaning given above and R₇ °, R₈ ° and R₉ ° have the meaning of R₇, R₈ and R₉, with the proviso that carboxyl and, if desired, free hydroxyl groups present in these radicals are protected by easily removable protective groups, implemented and, if necessary, existing protective groups are split off, or
• b) in a compound of the formula in which R, R₂, R₇ °, R₈ ° and R₉ ° have the abovementioned meaning, and R₅ represents an alkylidene or cycloalkylidene radical, splits off the oxazoline and dioxolane ring with acid, splits off any protective groups which may be present and into the optionally released amino group in 2- Position of the sugar molecule introduces the -CO-R residue, or
• c) a compound of the formula wherein R and R₂ have the meaning given above, and R₁ °, R₄ ° and R₆ ° represent an easily removable protective group, or a derivative thereof, with a compound of the formula wherein R₇ °, R₈ ° and R₉ ° have the meaning of R₇, R₈ and R₉, with the proviso that carboxyl and, if desired, free hydroxyl groups present in these radicals are protected by easily removable protective groups, condensed and any protective groups present are split off , or
• d) a compound of the formula wherein R, R₁ °, R₂, R₄ °, R₆ ° and R₇ ° have the meaning given above, with a compound of the formula in which R₈ ° and R₉ ° have the abovementioned meaning, with the proviso that carboxyl and, if desired, free hydroxyl groups in the radicals R₇ °, R₈ ° and R₉ ° are protected by easily removable protective groups, are condensed and any protective groups present are split off ,

and, if desired, one of the methods a) -d) Compound of formula I obtained with an acidic group in transferred a salt.   4. Use of a compound of general formula I according to Claim 1 or 2 or a pharmaceutically usable Salt of such a compound with an acidic group to stimulate the physical defense of humans and Animal.