DK154654B - METHOD OF ANALOGUE FOR THE PREPARATION OF IMMUNO MODULATING 2-OE2-ACYLAMINO-2-DESOXY- (D) -GLUCOSYL-3-OAA-ALKANCARBOXYL ACID Peptide DERIVATIVES OR PHARMACEUTICAL ACCEPTABLE ACCEPTED - Google Patents

Description

The invention relates to an analogous process for the preparation of novel immunomodulatory 2- [2-acylamino-2-deoxy- (D) -glucosyl-3-O] -alkanecarboxylic acid peptide derivatives of the general formula

wherein X is a carbonyl group, R is an alkyl group having from 2 to 17 carbon atoms or a phenyl group which is optionally substituted by alkyl of not more than 7 carbon atoms, R 1 is hydrogen, alkyl of not more than 7 carbon atoms or benzyl, R hydrogen or alkyl having not more than 7 carbon atoms, R 4 and R g means hydrogen or alkanoyl having 2 to 18 carbon atoms, R 7 means hydrogen, alkyl having not more than 7 carbon atoms, hydroxymethyl or phenyl, R 8 means carboxyl, carbamoyl, N-C 1-7 alkyl-carbamoyl, N-benzylcarbamoyl, N - (carbamoyl-methyl) -carbamoyl or

C 1-7 alkoxycarbonyl, and R 9 represents carboxyl, carbamoyl, N-C 1-7 alkylcarbamoyl or C 1-7 alkoxycarbonyl, the dogalkyl group R containing more than one carbon atom when R 2 represents methyl or when R 2 represents hydrogen and R or pharmaceutically acceptable salts of compounds with free carboxyl groups.

The process according to the invention is characterized in that a) is reacting a compound of the formula

wherein X and R have the meanings set forth above and R1, R40 and Rg ° represent the above groups R 1, R 4 and R 2 or each a readily cleavable protecting group, having a compound of the formula

wherein Z represents a reactively esterified hydroxy group, R 2 has the meaning given above and R 70, R0 O and R 9 ° have the meanings given above for groups R7, Rq and Rg, although carboxylic and free hydroxy groups present in these groups are readily protected leaving groups protecting groups, and optionally leaving groups protecting groups present, or b) condensing a compound of the formula \

wherein X, R and R 2 have the meanings given above and R 1 °, R 40 and R g ° have the meanings indicated for the groups R 1, R 4 and R 9 or each independently means a readily cleavable protecting group or a reactive acid derivative derivative having a compound of the formula

wherein R70, R8 ° and Rg ° have the meanings set forth above for the groups R7, Rg and Rg, however, these groups present carboxyl and, if desired, free hydroxy groups are protected with readily leaving protecting groups and leaving any protecting groups or groups present) condenses a compound of the formula

wherein R 1, R 1 °, R 2, R 40, R 9 ° and R 70 have the above meanings, with a compound of the formula

wherein Rg ° and Rg ° have the above meanings, wherein, however, in the groups R70, Rg ° and Rg ° carboxyl and, if desired, free hydroxyl groups are protected by readily cleavable protecting groups and cleave off any protecting groups present or d) in a compound of the formula

wherein R, R2, R70, R8 ° and Rg ° have the above meanings, and R5 represents an alkylidene or cycloalkylidene group, under acidic conditions, the oxazoline and dioxolane ring cleaves, and optionally cleaves any protecting groups or e) in association with formula

wherein the substituents have the meanings set forth above, but functional groups present, if desired, are protected with readily cleavable protecting groups, acylating the amino group with an acylating agent, such as introducer group R-X-, and optionally leaving any protecting groups or f) in a compound of the formula

wherein R Ri, R40 og and R R ° have the meanings set forth above for R₂, R4, and R eller respectively, or signifies an easily leaving group protecting group, R70,, Rg ° and Rg ° have the meanings given above for R7, Rg and Rg respectively, as carboxylic groups present in these groups and hydroxy groups may be protected with readily cleavable protecting groups, and X, R and R 2 have the meanings set forth above, however, with one compound of formula X, there is at least one readily cleavable protecting group, deprotecting the occurring protecting groups and, if desired, transferring one of the methods a ) to f) form a compound of formula I having at least one carboxyl group to a pharmaceutically acceptable salt thereof.

N-Acetyl-muramyl-L-alanyl-D-isoglutamine is the smallest structural unit which has the characteristic immune adjuvant properties of bacterial cell walls, cf. S. Kotani et al., BikenJ. 1975, 18 (2), 105-111 (Chemical Abstr. 83 (1975), 161981z). All of the experiments described in the above-mentioned literature on structural transformation of this structural unit, which is found in the bacterial cell walls, lead to compounds of no or weak effect.

Surprisingly, it has been found that the present synthetic compounds of the above Formula I and their acid addition salts have excellent adjuvant properties in association with diminished undesirable side effects.

The above-mentioned known compound, N-acetyl-muramyl-L-alanyl-D-isoglutamine, is herein referred to as 2-acetamido-3-O- <D1- [L1- (D1-carbamoyl-3-carboxypropyl) -carbamoyl- ethyl] -carbamoyl-ethyl> -2-deoxy-D-glucose and can also be referred to as 2- (2-acetamido-2-deoxy-D-glucose-3-O-yl) -D-propionyl-L-alanyl-D -isoglutamin.

The properties of this known compound are disclosed in DE Publication No. 2,718,010 to compounds which are also partly produced by the process of the present invention.

In Table I, Example 16, of the above-mentioned German publication specification, compound 1 on page 205 corresponds to the above-mentioned known compound used as a comparison compound, compound 1 on page 206 of said DE publication specification for the final product prepared following Example 5, compound 2 on page 206 corresponds to the final product made in the following example 2, compound 3 on page 206 corresponds to the end product prepared in the following example 37, compound 4 on page 206 corresponds to the final product prepared in the following example 39, compound 9 on page 206 corresponds to the final product made in the following example 39 following Example 7, and compound 1 on page 207 corresponds to the final product prepared in the following Example 4.

It is clear from Table I of said DE publication that all the compounds prepared according to the above examples have a surprisingly good effect over the known comparative compound. This superiority of the compounds in question is particularly evident by a comparison of the undesirable side effects. For example, it appears from Examples 17B, pages 210 and 211 of the above-mentioned DE disclosure that cats administered the known comparative compound at a dose of 1500 µg react with vomiting, diarrhea, depression, appetite deficiency and high fever, whereas administration of the compound prepared in the following Example 2 at the same dose does not cause any disruption to the well-being of the cats.

Thus, the compounds of the present invention differ in structure as well as in their overall effect from decanted compounds. Furthermore, the compounds of formula X wherein R 2 represents hydrogen are much easier to prepare as a result of the lack of asymmetry center than analogous compounds wherein R 2 is lower alkyl.

Alkyl is e.g. isopropyl, straight-chain or branched, non-substituted butyl, pentyl, hexyl or heptylog especially methyl, ethyl or n-propyl.

Alkanoyl of 2-18 carbon atoms is e.g. acetyl or propionyl.

Alkyl as a substituent in the above groups is primarily methyl or ethyl, but also n-propyl, isopropyl or straight or branched butyl.

Alkoxy is especially methoxy or ethoxy, moreover n-propoxy, isopropoxy, n-butoxy or iso-butoxy.

N-Alkoxycarbonyl is especially methoxycarbonyl or ethoxycarbonyl, but also n-propoxycarbonyl or isopropoxycarbonyl. Among the carbamoyl groups, the carbamoyl group itself is particularly preferred.

In the above compounds wherein R 2 represents an alkyl group, the R 2 -acetic acid amide group associated with the oxygen atom at the 3-position iglucosamine group is optically active, i.e. it is available in D form. When R7 does not hydrogenate, the Ry aminoacetic acid is in L-form.

The compounds of formula I, according to the nature of their substituents, are neutral or acidic compounds. Therefore, if acidic groups are present, they form salts with bases, such as ammonium salts or salts with alkali or alkaline earth metals, e.g. sodium, potassium, calcium or magnesium.

The compounds of the present invention exhibit valuable pharmacological properties, in particular a pronounced immunopotentiation effect. This can be shown using the test series described below.

1. Potentialization of cellular immunity in vivo:

Increase of late-type hypersensitivity to ovalbuminho guinea pigs.

Pirbright guinea pigs are immunized on day 0 with 10 mg ovalbumin complete Freund's adjuvant by injection of 0.1 ml of enantigenic adjuvant mixture in each of the two hind legs. 4 weeks later, skin reactions are triggered by intracutaneous injection of 100µg ovalbumin in 0.1 ml buffered physiological saline, and the reactions are quantified on the basis of the erythema surface and the skin thickness-calculated reaction volume 24 hours later. The antigen-specific increase in the reaction volume observed after 24 hours (late type reaction) applies as a measure of cellular agent immunity. Ovalbumin is a weak immunogen to induce alone or ivand 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") but must be applied for effective immunization in complete adjuvant to which are added mycobacteria (5 mg killed and lyophilized Mycobacterium butyricum per 10 ml "Bayol F" / "Arlacel A"). To demonstrate the immunopotentiation of test substances, dissenu instead of the mycobacteria at doses of 10 to 100 µg can be mixed in the antigen-oil mixture.

The present glucosamine peptides are capable of mimicking the action of the mycobacteria in the series of experiments described and quantitatively exceeding them.

Significant potentiation of the sentype activity overforovalbumin can also be achieved by the fact that compounds of the kind described are not incorporated into the antigen-oil mixture, but are subcutaneously administered at doses of 10 to 100 µg per animal over a few days after immunization (e.g. e.g., on day 0, 1, 2, 5, 6 and 7) in sodium chloride solution.

Thus, it is shown that compounds of the described species are capable of significantly increasing cellular immunity, both in admixture with the antigen itself (adjuvant effect is narrower) as well as by a separate delivery (systemic immune potentiation) with respect to time and place.

2. Potential humoral immunity in vivo:

Increase in antibody production to bovine serum albumin (BSA) in mice.

NMRI mice are immunized by intraperitoneal (i.p.) injection of 10 μg of precipitate-free BSA on days 0, 9, 15, and 29 days later, serum samples are taken, and their anti-BSA antibody content is assayed by passive hemaglutination technique. X the dose used is soluble BSA subimmunogenic for the animals, which receives it, ie. that it does not or only to a small extent trigger the production of antibody. Supplemental treatment of the mice with certain immunopotentiary agents before or after antigen administration leads to an increase in the antibody titre in the serum. The effect of the treatment is expressed by the obtained score value, i.e. by the sum of log 2 titer differences on the days of bleeding.

The present compounds are capable of significantly increasing antibody production to BSA significantly by five intraperitoneal or subcutaneous (s.c.) application of 100-300mg / kg / animal on five consecutive days (days 0 to 4) after immunization with BSA.

The immunostimulatory effect of said compounds, in contrast to the other bacterial immunoleptics (e.g., LPS from Escherichia coli), is antigen dependent. Injection of denye compounds, only in BSA-immunized, but not in non-immunized mice, results in an increase in the antirBSA titre. It is worth noting that subcutaneous administration of said compounds is as effective as intraperitoneal application, ie. that the observed immunoprotective effects are systemic and do not depend on the stimulant being administered along the same route as the antigen or having to mix with the antigen as is the case with classical adjuvants.

In the depicted experiments, it is shown that compounds of the kind described also manage to enhance the humoral immunity specific, that they improve the immunological sensitivity and that their immunopotentiating effects are due to the systemic activation of the immune apparatus.

3. Potentiation of humoral immunity in vitro: T-cell-substituting effect of antibody response in mouse spleen cells to sheep erythrocytes (SE).

To induce an antibody response, in many cases, defra-thymus-derived lymphocytes (T cells) are required. Dissection cells cooperate with the precursors of antibody-forming lymphocytes (B cells) and help them, by stimulation with so-called T-dependent antigens, respond to co-proliferation, differentiation and antibody synthesis. The spleen cell suspension from congenital athymic nu / nu mice contains functional T cells and is capable of e.g. in vitro presence of SE not to generate anti-SE antibodies. Surprisingly, the compounds in question are capable of functionally replacing T cells in such cultures and enabling an antibody response to SE. Addition of these substances to the now / now spleen cell cultures during the presence of SE leads to a significant increase in the number of antibody forming cells within 4 days.

The study results show that said compounds are capable of enhancing humoral antibody formation in vitro and to compensate for a defect in the T cell system.

4. Selective Mitogenicity for B Cells: Proliferative Effect in B Lymphocyte Cultures.

Suspensions of highly enriched B lymphocytes (lymph node cells from congenital athymic nu / nu mice) as well as extensive reneumature and mature T lymphocytes (thymus cells or cortisone resistant, i.e. 48 hours after a cortisone injection, persistent thymus cells from Balb / c mice) presence of the test substances. The incorporation of H3 thymidine into the lymphocytes during the last 18 hours of the culture period is the target of proliferation activity.

The compounds of this invention are mitogenic for B lymphocytes (i.e., for the precursors of the antibody-forming cells), but not for T lymphocytes. Thus, they are capable of stimulating the proliferation of lymphocytes participating in the humoral immune response.

5. Compatibility

For example, although compounds of the type described exert their potentiating effect on guinea pigs after a single dose of 0.05 mg / kg subcutaneously, on mice after application of 5 x 10 mg / kg subcutaneously, application of 5 x 300 mg / kg interperitoneally to the mouse ring is observed. toxic effects. The said substances therefore have an excellent therapeutic breadth.

The compounds of the invention may, on the one hand, by mixing with an antigen, increase the immunogenicity thereof, on the other, by systemic application, increase the immunological reactivity of the treated organism. Thus, said substances are capable of promoting both the cellular and the humoral immunity and to activate it. the pre-antibody formation responsible lymphocytes.

The new compounds can thus be used as adjuvants in admixture with vaccines to improve the vaccination result and to improve the infection protection mediated by humoral antibodies and / or cellular immunity overbacterial, viral or parasitic pathogens.

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

Furthermore, the new compounds can also be used without concomitant antigen administration to promote already below-threshold immune responses in humans and animals. Consequently, the compounds are particularly suitable for stimulation of the body's own defense, e.g. in chronic and acute infections or in selective (antigen-specific) immunological defects and in congenital, but also in the acquired (ie, non-antigen-specific) immunological defect conditions that occur in old age, in the course of serious primary diseases and, in particular, after therapy, medionizing rays or with immunosuppressive agents. acting hormones. Accordingly, said substances may preferably also be administered in combination with anti-infectious antibiotics, chemotherapeutics or other healing methods to counteract immunological damage. Furthermore, the compounds in question are also suitable for general prophylaxis of infectious diseases in humans and animals.

Particularly preferred are compounds of formula X wherein R 2 is hydrogen and the other groups have the above meanings and their salts.

In particular, compounds of formula I wherein R is alkyl or phenyl, R 1 is hydrogen or alkyl, R 2 is hydrogen or methyl, R 4 and R 9 are hydrogen, R 7 is hydrogen, alkyl or hydroxymethyl, R 9 is carbamoyl, and R 1 is carboxyl, however the alkyl group R contains more than 1 carbon atom if R 2 is methyl and their salts.

First of all, compounds of formula I wherein R is alkyl or phenyl, R 1 is hydrogen, R 2 is hydrogen or methyl, R 4 and R 5 are hydrogen, R 7 is hydrogen, methyl or hydroxymethyl, R 9 is carbamoyl, and R 9 is carboxyl. the alkyl group R contains more than 1 carbon atom if R 2 is methyl and their salts.

In process b) the condensation is carried out e.g. either by reacting the compound III in the form of the activated carboxylic acid with the amino compound IV, or at reacting the acid III with the compound IV, whose amino group is in activated form. For example, the activated carboxyl group may be an acid anhydride, preferably a mixed acid anhydride such as an acid azide, an acid amide such as etimidazolid, isoxazolid or an activated ester. Especially activated esters should be mentioned cyano methyl ester, carboxymethyl ester, p-nitrophenylthioester, p-nitrophenyl ester, 2,4,5-trichlorophenyl ester, pentachlorophenyl ester, N-hydroxysuccinimide ester, N-hydroxyphthalimide ester, 8-hydroxyquinolinester, -1-carboethoxy-quinoline ester, N-hydroxypiperidine ester or enol esters obtained with N-ethyl-5-phenyl-isoxazolium-3'-sulfonate. Activated esters may optionally also be obtained with etcarbodiimide with the addition of N-hydroxysuccinimide or enus substituted or e.g. with halogen, methyl or methoxy-substituted 1-hydroxy-benzotriazole, 3-hydroxy-4-oxo-3,4-dihydro-benzo [d] 1,2,3-triazine.

For example, the amino group is activated by reaction with etphosphite.

Among the methods of reacting with activated ester shells, in particular, such are mentioned with N-ethyl-5-phenyl-isoxazolium-3'-sulfonate (Woodward reagent K) or 2-ethoxy-1,2-dihydro-1-carboethoxy-quinoline or carbodiimide.

Easily cleavable protecting groups are those recognized by the peptide or sugar chemistry. For carboxyl group shells, tert-butyl, benzyl or benzhydryl and prehydroxy groups in particular acyl groups, e.g. lower alkanoyl groups such as acetyl, aroyl groups such as benzoyl and, above all, carbon dioxide derivatives such as benzyloxycarbonyl or lower alkoxycarbonyl, or alkyl, especially tert-butyl optionally with nitro, low alkoxy or halogen substituted benzyl or tetrahydropyranyl substituents or optionally substituted The 4- and 6 positions. Such alkylidene groups are in particular enlavalkylidene, primarily ethylidene, isopropylidene or propylidene group or also an optionally substituted, preferably substituted, benzylidene group.

These protecting groups can be cleaved in and in known manner. Thus, one can remove them hydrogenolytically, e.g. with hydrogen in the presence of a single-metal catalyst such as palladium or platinum catalyst, or by acid hydrolysis.

The starting materials used are known or can be produced in a manner known per se. Thus, one can obtain the compounds of formula III, e.g. by reacting the correspondingly in the 3-position unsubstituted sugars with enhalogen-R 2 -acetic acid, wherein R 2 has the above meaning and their esters, in the presence of a strong base.This is preferably halogen bromine or primarily chlorine.

In process c) the condensation is carried out e.g. either by reacting the compound V in the form of the activated carboxylic acid with the amino compound VI, or at reacting the acid V with the compound VI, whose amino group is in activated form. The activated carboxyl group can be, for example, an acid anhydride, preferably a mixed acid anhydride, an acid amide or an activated ester. As such, the above mentioned acid anhydrides, amides or esters in particular come into play. For example, the amino group is activated by reaction with a phosphite.

Also the readily cleavable protecting groups correspond to the dealers mentioned above. They can be cleaved in and in known manner, e.g. hydrogenolytic, for example, with hydrogen in the presence of a precious metal catalyst, such as palladium or platinum catalyst, or by acid hydrolysis.

The starting materials can be obtained in a manner known per se. You can, for example, sell the sole. in the manner shown above, react in the 3-position unsubstituted sugars with enhhalo-R 2 -acetamido-R 7 -acetic acid or a compound of formula III with an amino-Retic acid whose carboxyl groups are protected and the deprotected group.

In process a), a reactive esterified hydroxyl group is, in particular, a hydroxyl ester esterified ester group, preferably one esterified with hydrogen halides such as hydrochloric acid or hydrogen bromide or iodide.

The readily cleavable protecting groups are similar to the dealers mentioned above. They can be cleaved in and in known manner, e.g. hydrogenolytic, for example, with hydrogen in the presence of a precious metal catalyst, such as palladium or platinum catalyst, or by acid hydrolysis.

The starting materials used are known.

In method d) oxazoline and dioxolane ring acid are cleaved and any protecting groups present are cleaved.

In particular, the alkylidene R5 is the lower alkylidene such as the isopropylidene and the cycloalkylidene R5 is primarily the cyclopentylidene or cyclohexylidene.

The cleavage is carried out in a manner known per se, for example with an acidic ion exchanger, especially those with sulfonic acid groups such as "Amberlite" ® IR-120 (a styrene resin with strong acid 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. methanesulfonic acid, or an optionally substituted phenylsulfonic acid in the aromatic ring such as p-toluenesulfonic acid or trifluoroacetic acid. If you work in the presence of water, you get a free hydroxyl group in position 1. On the contrary, if one is working in the presence of an alcohol of the formula HO-R 1, which is an optionally substituted alkyl group, the 1-O-R 2 compound is obtained. Also, if one of the carboxyl groups Rg and / or Rg is esterified with an alcohol, especially a lower alkanol, it can be hydrolyzed with an aqueous acid, especially at higher temperature.

For example, the starting materials of formula II used in process e) can be prepared by cleavage of the dioxolane ring in a compound of formula IX.

In the compounds obtained, protecting groups of the peptide moiety can later be cleaved, e.g. by hydrogenolysis, such as with catalytically activated hydrogen, or by hydrolysis.

The starting materials thus used can e.g. is obtained by introducing a corresponding oxazoline, with a free hydroxyl group, to the 3-position of the sugar group, introduces the R2-acetamidopeptide group in one or more steps.

The compounds obtained with at least one carboxyl group can be transferred in their known manner to their salts, e.g.

by reaction of obtained acidic compounds with alkali or alkaline earth metal hydroxides.

The above-described processes are carried out in and in a manner known in the art, in the absence or preferably in the presence of diluents or solvents, when necessary under cooling or heating, under high pressure and / or in an inert gas atmosphere such as nitrogen atmosphere.

Taking into account all the substituents present, where necessary, especially in the presence of light hydrolyzable O-acyl groups, particularly gentle summer reaction conditions such as short reaction times, use of low acidic mild or basic agents, stoichiometric ratios, selection of suitable catalysts, solvents, and / or pressure conditions.

In the process of the invention, the starting materials may optionally be used in the form of a salt.

The present compounds can be used in the form of pharmaceutical preparations containing compounds of formula I.

The following examples illustrate the method according to the invention.

Example 1

A solution of 2.6 g of benzyl-3-O- <D1- [L1- (D1-carbamoyl-3-carboxy-propyl) -carbamoyl-ethyl] -carbamoyl-ethyl> -2-deoxy-2-propionylamino-αD -glucopyranoside benzyl ester in 67 ml60% fs acetic acid is hydrogenated with 0.6 g of 5% palladium-carbon catalyst at normal pressure and at room temperature (hydrogenation duration about 20 minutes).

The catalyst is filtered off and washed with a little 60% acetic acid, and the filtrate is evaporated to dryness in jet air vacuum. The residue is crystallized from ethanol / ether. There is thus obtained benzyl-3-O- [D1- [L-1- (D-1-carbamoyl-3-carboxy-propyl) -carbamoyl-ethyl] -carbamoyl-ethyl) -2-deoxy-2-propionylamino-aD -glucopyranoside with mp.155-160 ° C (decomposition) and optical rotation [α] §0 = + 105 ° ± 1 ° (dimethylformamide, c = 0.58).

The benzyl-3-O- <D1- [L1- (D1-carbamoyl-3-carboxy-propyl) -carbamoyl-ethyl] -carbamoyl-ethyl> -2-deoxy-2-propionylamino-α-D-glucopyranoside used as starting material benzyl ester can be obtained as follows: 4.1 g of benzyl-2-amino-4,6-O-benzylidene-3-O- (D1-carboxyethyl) -2-deoxy-α-D-glucopyranoside are dissolved in 200 3 ml triethylamine, and 0.95 ml of propionic acid chloride is added dropwise with stirring at room temperature. After 2 hours of stirring, the reaction mixture is evaporated to dryness and the residue is dissolved in 100 ml of water. Medical cold 2 N hydrochloric acid is adjusted to pH 4, the suction is separated off, washed with water and dried ivacuum over sodium hydroxide. The thus obtained benzyl-4,6-O-benzylidene-3-O- (D-1-carboxyethyl) -2-deoxy-2-propionylamino-α-D-glucopyranoside can be recrystallized from methanol, m.p. 257 ° C, [α] 25 D = + 132 ° ± 1 ° (dimethylformamide, c «1.086).

To a solution of 3.1 g of the obtained compound in 150 ml of acetonitrile and 0.9 ml of triethylamine is added 1.5 g of N-ethyl-5-phenyl-isoxazolium-3'-sulfonate (Woodwards reagent K) and stirred at room temperature. until complete dissolution (about 60 minutes). 2.2 g of L-alanyl-D-glutamic acid-1-amide-γ-benzyl ester hydrochloride and 0.9 ml of triethylamine are now added and stirred at room temperature for an additional 18 hours. After distilling off the solvent in water-jet vacuum, water is added to the residue from which it is insoluble, washed thoroughly with water and dried. The obtained benzyl-4,6-O-benzylidene-3-O- <D1- [L1- (D1-carbamoyl-3-carboxypropyl) -carbamoyl-ethyl] -carbamoyl-ethyl) -2-deoxy-2-propionylamino-aD -glucopyranoside benzyl ester of Rf value 0.45 on silica gel thin-layer plates of the methylene chloride / methanol 10/1 system is dissolved in 200 ml of 60% acetic acid and left for 1.5 hours at 95-100 ° C. After cooling, the solvent is distilled off. The residue is taken up 2 more times in a little water and evaporated each time to dryness. The obtained benzyl-3-O- <D1- [L1- (D1-carbamoyl-3-carboxy-propyl) -carbamoyl-ethyl] -carbamoyl-ethyl> -2-deoxy-2-propionylamino-α-D-glucopyranoside benzyl ester crystalline From methanol, m.p. 189 ° C, [α] 21 D = + 100 ° ± 1 ° (dimethylformamide, c = 1.268).

Example 2

A solution of 4 g of benzyl-2-acetamido-3-O - [[1- (D1-carb-amoyl-3-carboxy-propyl) -carbamoyl-ethyl] -carbamoyl-methyl} -2-deoxy-α-D-glucopyranoside -Benzyl ester in 80 ml of methanol is hydrogenated with 0.4 g of 5% palladium coal as a catalyst at normal pressure and at room temperature until stationary. The catalyst is filtered off, washed with a little methanol and the filtrate is evaporated to dryness in water-jet vacuum. The residue is dissolved in 50 ml of distilled water and further hydrogenated with 1 g of 5% palladium-carbon as catalyst at normal pressure and room temperature until stagnant. The catalyst is filtered off, then washed with a little water and the filtrate is evaporated to dryness. The obtained 2-acetamido-3-O - <[L1- (D1-carbamoyl-3-carboxy-propyl) -carbamoyl-ethyl] -carbamoyl-methyl) -2-deoxy-D-glucose dry vacuum in high vacuum over phosphorus pentoxide, [a ] §® = + 10 ° ± 1 ° (water, c - 0.930).

The starting material can be prepared as follows:

To a solution of 9.5 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O-carboxymethyl-2-deoxy-α-D-glucopyranoside in 400 ml of acetonitrile and 3 ml of triethylamine is added 5.3 g of N -ethyl-5-phenyl-isoxazolium-3'-sulfonate (Woodward's reagent K) and stirred at room temperature until a clear solution is obtained. 7.15 g of L-alanyl-D-glutamic acid 1-amide-Y-benzyl ester hydrochloride, 3 ml of triethylamine and 200 ml of acetonitrile are then added and the reaction mixture is stirred for an additional 18 hours at room temperature. The crystallized benzyl-2-acetamido-4,6-O-benzylidene-3-O - [[1- (D1-carbamoyl-3-carboxy-propyl) -carbamoyl-ethyl] -carbamoyl-methyl> -2-deoxy-benzylidene α-D-glucopyranoside benzyl ester is sucked off, washed with semi-saturated sodium bicarbonate solution and water dried, [<x] §0 = + 81 ° ± 1 ° (dimethylformamide, c = 0.816).

A solution of 8 g of this compound in 400 ml of 60% acetic acid is left at 80 ° C for one hour. After cooling, the solution is evaporated to dryness and to the residue is added twice more 50 ml of water and evaporated each time to dryness. The crystalline residue obtained is stirred with a little water and the crystals are sucked off and dried. There is thus obtained benzyl-2-acetamido-3-O - <[L-1- (D-1-carbamoyl-3-carboxy-propyl) -carbamoyl-ethyl] -carbamoyl-methyl> -2-deoxy-α-D-glucopyranoside benzyl ester with m.p. 200-202 ° C, [α] 20 = + 77 ° ± 1 ° (dimethylformamide, c = 0.599).

Example 3

A solution of 0.9 g of benzyl-2-acetamido-3-O- [L1- (D1-carb-amoyl-3-carboxy-propyl) -carbamoyl-ethyl] -carbamoylmethyl-2-deoxy-6-o-steaoyl -aD-glucopyranoside benzyl ester in 40 ml of methanol is hydrogenated with 0.2 g of 5% palladium carbon as catalyst at normal pressure and room temperature until 22.4 ml of hydrogen is taken up. The catalyst is filtered off, washed with methanol and the filtrate is evaporated to dryness to give benzyl-2-acetamido-3-O- [L- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoyl-methyl-2 deoxy-6-O-stearoyl-α-D-glucopyranoside, [α = + 33 ° ± 1 ° (chloroform, c = 1.046).

The starting material can be prepared as follows: benzyl-2-acetamido-3-O- [L1- (D1-carbamoyl-3-carboxypropyl) - carbamoylethyl] -carbamoylmethyl-2-deoxy-α-D-glucopyranoside-benzy 1 ester x 30 ml pyridine with stirring and exclusion humidity drop dropwise a solution of 1.4 g of stearic acid chloride 7.5 ml of methylene chloride over one hour and stir for 48 hours at room temperature. The reaction mixture is poured into ice water and extracted with methylene chloride. The inorganic phase is washed with ice-cold 2N hydrochloric acid and water, dried over magnesium sulfate and evaporated to dryness. The residue is purified by column chromatography on silica gel eluting with ethyl acetate.

There is thus obtained benzyl-2-acetamido-3-O- [L1- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoyl-methyl-2-deoxy-6-o-stearoyl-α-D-glucopyranoside benzyl ester with [ α] §0 = + 30 ° ± 1 ° (chloroform, c = 1.203).

Example 4

Analogously to Example 2, benzyl-2-acetamido-4,6-O-benzylidene-3-O- (D1-carboxy-propyl) -2-deoxy-α-D-glucopyranoside is condensed with L-alanyl-D-glutamic acid-1- amide-γ-benzyl ester hydrochloride and cleaves the protecting group. There is thus obtained 2-acetamido-3-O- (D-1- [L-1- (D-1-carbamoyl-3-carb-oxy-propyl) -carbamoyl-ethyl] -carbamoyl-propyl> -2-deoxy-D-glucose.

The starting material can be prepared as follows:

To a solution of 60 g of benzyl-2-acetamido-4,6-O-benzylidene-2-deoxy- <xD -glucopyranoside in 600 ml of dimethyl formamide is added 10 g of sodium hydride and stirred for 1.5 hours in a nitrogen atmosphere at 45 ° C. After cooling to 0 ° C, 75 ml of D, L-α-bromobutyric acid ethyl ester is added. The reaction mixture is stirred for one hour at room temperature and one hour at 50 ° C, neutralized with acetic acid and evaporated the solvent in water jet vacuum. The residue is partitioned between methylene chloride and water, washed again the inorganic phase with water, dried with magnesium sulfate and evaporated the solvent. The column chromatographic column is dried in high vacuum, dried on silica gel. Eluting with methylene chloride / ethyl acetate (85:15) gives benzyl-2-acetamido-4,6-O-benzylidene-3-O- (D1-carboxy-propyl) -2-deoxy-o: -D-glucopyranoside ethyl ester with [α = + 113 ° to 1 ° (chloroform, c = 0.5), m.p. 154 ° C (from methylene chloride / ether) and Rf value 0.21 and benzyl-2-acetamido-4,6-O-benzylidene-3-O- (L1-carboxy-propyl) -2-deoxy-aD -glucopyranoside ethyl ester with [α = + 42 ° ± 1 ° (chloroform, c = 0.511), m.p. 240 ° C (from ethyl acetate) and Rf value 0.04.

To a solution of 38.1 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O- (D1-carboxy-propyl) -2-deoxy-α-D-glucopyranoside ethyl ester in 300 ml of methanol is added. 100 ml of 1 N sodium hydroxide solution, and the mixture is left for one hour at 60 ° C. Then the solution is cooled, evaporated to ca. 150 ml, dilute with 400 ml of ice water and add 100 ml of 1 Niskold hydrochloric acid. The crystallized product is extracted, washed with water and dried. Benzyl-2-acetamido-4,6-O-benzylidene-3-O- (D-1-carboxy-propyl) -2-deoxy-α-D-glucopyranoside is obtained. 210-213 ° C and [a] 2® = + H0 ° ± 1 ° (dimethylformamide, c = 0.554).

In an analogous manner, benzyl-2-acetamido-4,6-O-benzylidene-3-O- (L-1-carboxy-propyl) -2-deoxy-α-D-glucopyranoside is obtained, m.p. 285 ° C and [α] β0 = + 71 ° ± 1® (dimethylformamide, c = 0.589).

Example 5 3.77 g of 2-phenyl-4,5- [3-O- (D1-carboxyethyl) -5,6-O-isopropyl ylidene-D-glucofuranoJ-A 2 -oxazoline in 60 ml of acetonitrile and 15 ml of dimethyl formamide is stirred 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, whereby almost everything dissolves. Then mn adds 3.44 g of L-alanyl-D-glutamic acid-1-amide-7-benzyl ester hydrochloride and an additional 1.4 ml of triethylamine and stirs for 24 hours at room temperature. Evaporate the oil pump vacuum to a syrup and chromatograph on silica gel with a mixture of chloroform and acetone (8: 2). One obtains a colorless solid syrup which crystallizes by rubbing with ether. Mp. 96-99 ° C, [α] 25 D = + 13 ° (in chloroform).

The crystalline benzyl ester is hydrogenated with 5% palladium carbon in dioxane at room temperature and normal pressure and then evaporated in vacuo the corresponding acid as syrup. It is stirred in water with 10 ml of Dowex® 50-H + boiling temperature for 15 hours. . After filtration and freeze-drying, a colorless powder with decomposition point 140 DEG C. is obtained. The obtained 2-benzoylamino-3-O- <D1- [L1- (D1-carbamoyl-3-carboxy-propyl) -1-carbamoyl-ethyl] -carbamoyl -ethyl> -2-deoxy- α, β -D-glucose contains, depending on the drying conditions, varying amounts of crystal water, in the above case post-drying at 60 ° C, 0.01 mm Hg, for 15 hours: 1/3 water.

Example 6 б, 0 g of 2-phenyl-4,5- [3-O- (D1-carboxypropyl) -5,6-O-isopropyl ylidene-D-glucofurano] -A 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 dimethyl formamide for one hour at 0-5 ° C to dissolve everything. Then 5.55 g of L-alanyl-D-glutamic acid-1-amide-7-benzyl ester hydrochloride is added and an additional 2.35 ml of triethylamine is added and stirred for 48 hours at room temperature. Evaporate in oil pump vacuum and chromatograph on silica gel in a mixture of chloroform and ethanol (19: 1). 9 g of the colorless syrup thus obtained is hydrogenated in dioxane with 5% palladium carbon, filtered off the catalyst, evaporated in vacuo and hydrolyzed in a single mixture of 40 ml of tetrahydrofuran and 30 ml of water with 1.5 ml of trifluoroacetic acid at room temperature. Then, the man water evaporates in vacuo four times to dryness, dissolves in water and lyophilizes. The obtained 2-benzoylamino-3-O- <D1- [L1- (D1-carbamoyl-3-carboxy-propyl) -carbamoyl-ethyl] -carbamoyl-propyl> -2-deoxy-α, β-D-glucose crystallizes with 0.5 mol water, m.p. 114-152 ° C, [α] S + + 17 ° (in methanol).

Example 7 3.63 g of 2-phenyl-4,5- [3-O-carboxymethyl-5,6-O-isopropylidene-D-glucofurano] -O 2 -oxazoline, 3.43 g of L-alanyl-D-glutamic acid 1-Amide-Y-benzyl ester hydrochloride, 1.21 g of N-hydroxysuccin imide, 2.16 g of dicyclohexylcarbodiimide and 1.45 ml of triethylamine are dissolved in 40 ml of dimethyl formamide and stirred for 24 hours at room temperature. Evaporate in oil pump vacuum, take up dichloroethane and water, extract the precipitated dicyclohexyl urea and equip the organic phase twice with water and the aqueous phase twice with dichloroethane. After drying and evaporation of the organic phases, a solid syrup is obtained which is chromatographed on silica gel with a mixture of chloroform / ethanol (9: 1). The obtained peptide ester which crystallizes by rubbing with ether melts at 167-168 ° C, [α] δ ° = "5 ° (chloroform).

4.5 g of said ester is hydrogenated in dioxane with 5% spalladium carbon, the catalyst is filtered off and extracted with ethanol. The combined filtrates are evaporated and the residue is recrystallized from isopropyl alcohol. The acid obtained melts at 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 evacuated to dryness. Rubbing with ether gives a colorless powder, 2-benzoylamino-3-O - <[L- (D1-carbamoyl-3-carboxy-propyl) -1-carbamoyl-ethyl] -carbamoyl-methyl) -2-deoxy-α , (SD-glucose, mp 175-177 ° C (as hydrate)).

By variation of that in Acta Chem. Scand. 18, 185 (1964), the starting material can be prepared as follows: 100 g of 2-phenyl-4,5- [5,6-O-isopropylidene-D-glucofurano] -δ2-oxazoline are dissolved in the exclusion of moisture and carbon dioxide for 1 hour. per liter of acetonitrile and 15.2 g of a 55% NaH mineral oil dispersion are added portionwise with good stirring which is continued for one hour at room temperature. Then, at 0 ° C, 42 ml of chloroacetic acid ethyl ester is added dropwise and after 1.5 hours another 42 ml. After 1.5 hours, 11.4 gNaH dispersion is added again, stirred for half an hour and dripped at 0 ° C. ml of chloroacetic acid ethyl ester. After another 2 hours, allow to warm to room temperature and evaporate the evacuum, finally in the oil pump vacuum, to a syrup. This was taken up with ether, extracted 3 times with water, dried in the drying phase over sodium sulfate and, after evaporation, 155 gave a brown oil. This is dissolved in 150 ml of methanol and a solution of 30 g of potassium hydroxide is added in 150 ml of water, extracted twice with ether and the ether phase is washed once with water. The aqueous phases are freed from ether in vacuo and adjusted with 1 N hydrochloric acid with pH meter to pH 4.

The precipitated crystalline 2-phenyl-4,5- [3-O- (carboxymethyl-5-yl) -5,6-O-isopropylidene-D-glucofuranoJ-A 2 -oxazoline is sucked off, washed with water and dried. 107 g, 93% of this theoretical are obtained, m.p. 186-188 ° C, [a] §0 = -9 ° (CHCl3, c = 0.9) and + 23 ° (CHCl3, c = 3).

Example 8 2.5 g of 2-benzamido-2-deoxy-3-O- [L1- (D1-carbamoyl-3-carb-oxy-propyl) -carbamoylethyl] -carbamoylmethyl-D-glucopyranose are dissolved in a mixture of 17 ml of absolute pyridine and 4 ml butyric anhydride. After 20 hours at room temperature and 2 hours at 50 ° C, water is added and evaporated in vacuo. The residue is dissolved in chloroform, extracted with 1N hydrochloric acid and water, dried over sodium sulfate and evaporated the chloroform phase. The oily residue is extracted several times with ether to give a solid amorphous mass of m.p. 110-120 ° C, α, β, 1,4,4-tributyroyl-2-benzamido-2-deoxy-3-O- [ L- (D1-carbamoyl-3-carboxy-propyl) -carbamoylethyl] -carbamoylmethyl-D-glucopyranose, Rf = 0.52 in CHCl3 CH2OH = 7: 2 (silica gel layer, Merck).

Example 9 2.3 g of 2-benzamido-2-deoxy-3-O- [L1- (D1-carbamoyl-3-carb-oxy-propyl) -carbamoylethyl] -carbamoylmethyl-D-glucopyranose dissolve in 20 ml of pyridine and 5 ml acetic anhydride. After 3 hours at room temperature, water is added and evacuated is evaporated. Purify on 70 g of silica gel with CHCl3 / CH3 OH = 3: 1 to give a colorless powder with m.p. 122-158 ° C = + 48 ° (CH 3 OH, c = 1.074), α, 3- [1,4] triacetyl-2-benzamido-2-desoxy-3-O- [Ll- (Dl -carbamoyl-3-carboxypropyl) -carbamoyl-ethyl] -carbamoylmethyl-D-glucopyranose, R f = 0.54, CHCl 3: CH 3 OH = 3: 2 (silica gel thin layer, Merck).

Example 10 6.33 g of 2-phenyl-4,5- (3-O-carboxymethyl-5,6-O-isopropylidene-D-glucofurano-Δ-oxazoline, 5.75 g of 2-ethoxy-N-carbethoxy) 1,2-dihydroquinoline (EEDQ) and 9.3 ml of triethylamine are added to the solution of the trifluoroacetate of L-alanyl-D-glutamic acid dibenzyl ester (obtained from 8.3 g of N-tert-butoxycarbonyl-L-alanyl-D-glutamic acid dibenzyl ester with 5.1 ml of trifluoroacetic acid and 2.6 ml of dichloroethane (4 hours of hydrolysis at 40 ° C) in 70 ml of dichloroethane The reaction mixture is reacted for 15 hours at 40 ° C, diluted with chloroform, quenched twice with water and the aqueous phases once with chloroform. drying over sodium sulfate and evaporation of the chloroform solution gives 19.9 g of a single oil which is purified over 400 g of silica gel (Merck) by elution with ether, then with chloroform / acetone (17: 3). [3-O- (1L- <1D, 3-dibenzyloxycarbonyl-propyl> -carbamoyl-ethyl) -carbamoylmethyl-5,6-O-iso-propylidene-D-glucofurano JA-oxazoline, mp 113-116 ° C and [αβ ° = -47 ° (CHCl3, c = 1.54).

7 g of the above compound are hydrogenated with 1.8 g of 5% spalladium coal in a mixture of 80 ml of tetrahydrofuran and 20 ml of water to a standstill, from which the catalyst is suctioned, evaporated in vacuo and rubbed with ether. 4.9 g of the dicarboxylic acid is thus obtained as a colorless powder.

4.4 g of the above dicarboxylic acid is stirred for 20 hours at 40 ° C with 11 ml of the "Dowex-50" ® W x 4 ion exchanger in a single mixture of 45 ml of tetrahydrofuran and 20 ml of water. After-filtration and clarification with charcoal "Darco G 60", freeze-dry the man solution and obtain colorless, amorphous 2-benzamido-2-deoxy-3-O- [1L- (1D, 3-dicarboxy-propyl) -carbamoyl-ethyl] -carb. amoylmethyl-D-glucopyranose with the optical rotation [<x] 3 ° - + 25 ° (Η · ρ, c = 0.997).

Example 11

Analogously to Example 10, 5.7 g of 2-phenyl-4,5- [3-O-carboxymethyl-5,6-O-isopropylidene-D-glucofurano] -δ2-οχ-azoline are condensed with 4.9 g of L- seryl-D-glutamic acid oleamide-7-tert-butyl ester hydrochloride with the addition of 2.3 ml of triethylamine and 5.2 g of 2-ethoxy-N-carbethoxy-1,2-dihydroquinoline in 45 ml of dichloroethane. After 18 hours at 40 ° C, one crystallized. An additional 50 ml of dichloroethane is added, cooled in ice, suctioned off and the crystals are washed with cold dichloroethane. The assays, colorless crystals of 2-phenyl-4,5- [3- (1- L - <1 -D-c ar bamoy1-3-1ert-butoxycarbony1-propyl) -carbamoyl-2-hydroxyethyl) -carbamoylmethyl -5,6-O-iso-propylidene-D-glucofuranoJ-A 2 -oxazoline has a snip. 187-188 ° C and [agO = + 7 ° (CH + DH, c = 1.125).

2 g of this compound is hydrolyzed at room temperature for 20 hours with a mixture of 15 ml of methylene chloride and 5 ml of trifluoroacetic acid. Evaporate in oil pump vacuum, rubbing residue with ether to give 2-benzamido-2-deoxy-3-O- [1- [1- (1-carbamoyl-3-carboxypropyl) carbamoyl-2-hydroxyethyl] carbamoylmethyl-D-glucopyranose a beige powder with mp.100-115 ° C, [α] g ° = + 23 ° (H #, c = 0.886) crystallizing with 2 moles of water and 1 mole of trifluoroacetic acid.Rf = 0.28, CHCl : 1 (silica gel thin layer, Merck).

Example 12

Analogously to Example 10, 5.25 g of 2-phenyl-4,5- [3-O-carboxymethyl-5,6-O-isopropylidene-D-glucofuranooxazoline and the trifluoroacetic acid salt of L-alanyl-D-glutamic acid are condensed. -an-propylamide-y-benzyl ester, obtained from 6.2 g of N-tert-butoxy-carbonyl-L-alanyl-D-glutamic acid-an-propylamide-y-benzyl ester and 4.2 ml of trifluoroacetic acid in 2.5 ml of dichloroethane , 6 hours at 40 ° C in 60 ml of dichloroethane supplementation of 7.75 ml of triethylamine and 4.8 g of 2-ethoxy-N-carbethoxy-1,2-dihydroquinoline. After 20 hours at 40 ° C, dilute with 50 ml of chloroform, shake twice with water and twice with chloroform. After drying and evaporation of the chloroforph ash, 15 g of an oil is obtained which is purified over 200 g of silica gel (Merck) by elution with ether, then with chloroform / acetone = 7: 3. 6.4 g of a colorless amorphous substance are obtained with Rf = 0.35, CHCl3: acetone = 7: 3 (silica gel layer, Merck). This is hydrogenated with 1.8 g of 5% palladium coal in 80 ml of tetrahydrofuran and 20 ml of water, filtered from the catalyst and evaporated. Oxygen indicator Rf = 0.58, CHCl3 / CH2OH = 3: 1 (silica gel layer,

Merck). Then, with 10 ml of the ion exchanger "Dowex50" ® w x 4, 50 ml of tetrahydrofuran and 25 ml of water are stirred for 15 hours at room temperature and for 12 hours at 40 ° C. After filtration, clarification with charcoal "Darco-G-60", again filtration and freeze-drying, the colorless, amorphous 2-benzamido-2-deoxy-3-O- [1-L- (D1Nn-propyl-carbamoyl-3- carboxy-propyl) -carbamoylethyl] -carbamoyl-methyl-D-gluco-pyranose, m.p. 65-140 ° C and [α] β0 = + 28 ° (water, c = 1.03), Rf = 0.48, CHCl3: CH3OH = 1: 1 (silica gel layer, Merck).

Example 13 7.3 g of 2-phenyl-4,5- [3-O-carboxymethyl-5,6-O-isopropylidene-D-glucofurano] -O 2 -oxazoline, 6.5 g of α-amino-isobutyroyl-D glutamic acid α-amide-7-tert-butyl ester hydrochloride and 2.9 g of chloromyric acid isobutyl ester are dissolved in 25 ml of dimethylformamide and 50 ml of dichloroethane. A solution of 6.1 ml of triethylamine in 20 ml of dichloroethane is added dropwise at -15 to -10 ° C over 30 minutes. Then allow to warm to room temperature and stir for another 15 hours at room temperature. Manifold with 50 ml of dichloroethane, extract with water, two times with 0.5 N NaOH and three times with water, the aqueous phase two times with dichloroethane, dry the organic phases and few evaporation of 16.6 g of oil. This is purified on silica gel (Merck) by elution with CHCl3: C2H50H = 19: 1. 9.7 g of colorless amorphous 2-phenyl-4,5- <3-O- [1-methyl-1- (1-D-carb-amoyl-3-tert-butoxycarbonyl-propyl) -carbamoylethyl] -5 , 6-O-isopropylidene-D-glucofurano) -O 2 -oxazoline with optical rotation: [α = + 6 ° (CHCl 3, c = 1.027), m.p. 75-89 ° C, Rf = 0.35, CHCl3: C2H50H = 9: 1 (silica gel layer, Merck).

8.3 g of the above compound are allowed to stand for 15 hours at room temperature in a mixture of 20 ml of trifluoroacetic acid, 60 ml of methylene chloride and 2 ml of water. Then evaporate in vacuo and rub the residue with ether. The resulting rose-colored powder is dissolved in 200 ml of water and clarified with 0.5 g of charcoal "Darco G 60". After filtration and evaporation, the colorless amorphous 2-benzamido-2-deoxy-3-0- [1-methyl-1] is obtained. - (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl-D-glucopyranose, mp 110-120 ° C, [a] g ° = + 31 ° (H +, c = 0.88), Rf = 0, 52, acetone / ethanol = 1: 1 (silica gel thin layer, Merck) crystallizing with 0.6 mole of trifluoroacetic acid and 1.7 mole water.

Example 14

Analogously to Example 10, 2-benzamido-2-deoxy-3-O-carboxymethyl-3-ethyl-D-glucopyranoside and the trifluoroacetic acid salt of L-alanyl-D-glutamic acid α-ainide-Y tert-butyl ester with 2-ethoxy-N-carbethoxy-1,2-dihydroquinoline corresponding glycopeptide with [a] 2 °? -23 ° (CH 2 pH, c = 1,107) and R f = 0.47 (CH 2 Cl 2: C 2 H 50 H = 8: 2) and after hydrogenation with 5% fs palladium carbon in tetrahydrofuran / water 2-benzamido-2-deoxy 3-O- [L1- (D1-carbamoyl-3-carboxyprop-yl) -carbamoylethyl] -carbamoylmethyl-O-ethyl-D-glucopyranoside, m.p. 215-217 ° C, [α] g ° = -22 ° (CH $) H, c = 0.97), R f = 0.36 in CHCl 3: CH 3 OH = 1: 1 (silica gel thin layer, Merck).

The 2-benzamido-2-deoxy-3-O-carboxymethyl-3-ethyl-D-glucopyranoside used as the starting material is obtained as follows: 2-Phenyl-4,5- [3-O-carboxymethyl-5,6- O-isopropylidene-D-gluco-furano] -2-oxazoline is dissolved in 0.1 N HCl / C 2 H 5 OH and left for 6 hours at room temperature. The sodium ethylate is neutralized in ethanol, evaporated to dryness and with acetone. The solution is filtered over a layer of silica gel (Merck), the eluate is evaporated to dryness and the residue is extracted twice with ether at room temperature. Post-crystallization from ethyl acetate gives 2-benzamido-2-des-oxy-3-O-carbethoxymethyl- / 3-ethyl-D-glucopyranoside, m.p.185-188 ° C and [a] 2 ° = -35 ° (CH φΗ , c = 1.121).

9.4 g of this ester is hydrogenated with a solution of 1.7 g of potassium hydroxide in 250 ml of ethanol and 25 ml of water for 2 hours at room temperature. Then, adjust the pH with 1 N hydrochloric acid to 3.5 and evaporate in vacuo. Rub the residue first with ether, then 3 times, each time with 20 ml of water, and suction. There are thus obtained crystals of 2-benzamido-2-deoxy-3-O-carboxymethyl- / 3-ethyl-D-glucopyranoside, m.p. 205-210 ° C and [a] 2 ° = -40 ° (CH ΗΗ, c = 1.04).

Example 15 2-Benzamido-2-deoxy-3-O- [(D1-carbamoyl-3-carboxy-propyl) -carbamoylmethyl] -carbamoylmethyl-D-glucose gives 3 g2-phenyl-4,5- [3- O - <(D1-carbamoyl-3-carboxypropyl) -carbamoylmethyl> -carbamoylmethyl-5,6-O-isopropylidene-D-gluco-furano JA 2 -oxazoline by hydrolysis with 1.5 ml of trifluoroacetic acid in a mixture of 15 ml of dimethoxyethane and 15 ml of water at 40 ° C for 3 hours. Evaporate in vacuo to dryness and extract the residue again with ether. The residual powder is dissolved in water and treated with charcoal "Darco G 60", filtered and lyophilized. There is thus obtained a colorless amorphous substance with m.p. 115-155 ° C and [a] 2® = + 34 ° (water, c = 0.81), Rf = 0.28, CHCl 3: CH 3 OH = 1: 1 (silica gel layer, Merck).

The starting material is obtained as follows: 8.0 g of N-tert-butoxycarbonyl-glycyl-D-glutamic acid α-amide-γ-benzyl ester are dissolved in a mixture of 6.3 ml of trifluoroacetic acid and 7 ml of dichloroethane and react in 2 days at room temperature and 3 hours at 45 ° C. In addition, 12.1 ml of triethylamine, 7.0 g of 2-ethoxy-N-carbethoxy-1,2-dihydroquinoline (EEDQ) and 8.1 g of 2-phenyl-4,5- [3-O-carbene] are added under cooling. oxymethyl-5,6-O-isopropylidene-D-glucofurano JA 2 -oxazoline and 20 ml of dimethylformamide. After 20 hours at 40 ° C, the evaporator is evaporated in an oil vacuum and the residue distributes between methylene chloride and water. After drying and evaporation of the methylene chloride phase, a solid residue is obtained, extracted twice with ether and recrystallized from fratoluene. Yield 8.25 g, m.p. 1576C, [a] 3® = + 10 ° (CHCl 3 3. c = 1.48), R f = 0.35 (CHCl 3: ethanol = 9: 1) (silica gel layer, Merck).

The benzyl ester thus obtained is hydrogenated with 1 g of 5% spalladium carbon in 100 ml of tetrahydrofuran and 25 ml of water as the condition is achieved. After filtering off the catalyst and evaporation, the substance is chromatographed on 250 g of silica gel (Merck) in CHCl3 / CH2 OH = 4: 1. 5.8 g of colorless amorphous 2-phenyl-4,5- [3-O - <(D1-carbamoyl-3-carboxypropyl) -carbamoylmethyl> -carbamoylmethyl-5,6-O-isopropylidene-D-glucofu -rano JA 2 -oxazoline with R f = 0.43, CHd ^ CHOOH = 3: 2 (silica gel layer, Merck).

Example 16

Analogously to Example 10, 9.5 g of 2-phenyl-4,5- [3-O-carboxymethyl-5,6-O-isopropylidene-D-glucofurano \ -δΡ-oxazoline are condensed with 6.25 g of L-alanyl D-glutamic acid α, γ-diamide hydrochloride with the addition of 3.4 ml of triethylamine and 7.95g of 2-ethoxy-N-carbethoxy-1,2-dihydroquinoline (EEDQ) in a single mixture of 50 ml of dichloroethane and 150 ml of dimethyl formamide The mixture is reacted with stirring for 2 days at room temperature and 4 hours at 40 ° C. Evaporate in the oil pump vacuum and extract the residue, first twice with ether, then twice with ice water. After drying, the product can be recrystallized from dichloroethane. Colorless crystals are thus obtained with m.p. 170-184 ° C, [α] 20 D = + 3 ° (DMSO, c = 1.43), R f = 0.64, CHCl 3: CH 3 OH = 3: 1 (silica gel thin layer, Merck).

6.1 g of this compound are hydrolyzed with 13.5 ml of the "Dowex 50" ® exion exchanger in a mixture of 60 ml of dimethoxyethane and 60 ml of water for 15 hours at room temperature. After filtration and evaporation, the residue is taken up with water, clarified with coal "Darco -G-60 ", suction off and freeze-drying filtrate. Colorless amorphous 2-benzamido-2-deoxy-3-O- [L-1- (D-1,3-dicarbamoylpropyl) -carbamoylethyl] -carbamoyl-methyl-D-glucopyranose is obtained. 82-143 ° C, [α] 3 ° = + 24 ° (H +, c = 0.98), Rf = 0.45, CHCl3: CH3 OH = 1: 1 (silica gel thin layer, Merck). The substance crystallizes with 1.23 molar crystal water.

Example 17

Analogously to Example 16, ethyl 2-benzamido-2-des-oxy-3-O-carboxymethyl-O-D-glucopyranoside and L-alanyl-D-glutamic acid α-7-bis-methylamide hydrochloride are obtained | 3 -ethyl-2-benzamido-2-deoxy-3-O- [L1- (D1,3-bis-N-methyl-carbamoyl-propyl) -carbamoylethyl] -carbamoylmethyl-D-glucopyranoside m.p. 233-240 ° C, [α] g ° = -20 ° (CH $> H, c = 0.937), Rf = 0.38 iCHCl 3: ethanol 7: 3 (silica gel, thin-layer plates, Merck).

Example 18

Analogously to Example 16, 2-benzamido-2-deoxy-3-O-carboxymethyl-D-glucopyranose 2-benzamido-2-deoxy-3-O- [Ll- carbamoyl-propyl) -carbamoyl-ethyl] -carbamoylmethyl-D-glucopyranose, m.p. 125-132 ° C, [α] g ° = + 24 ° (H +), c = 0.93), Rf = 0.26 CHCl3: ethanol = 7: 3 (silica gel, thin-layer plates, Merck).

Example 19

Analogously to Example 16, 2-benzamido-2-deoxy-3-O-carboxymethyl-D-glucopyranose and the hydrochloride of L-alanyl-D-glutamic acid dimethyl ester 2-benzamido-2-deoxy-3-O [11- (D1,3-bis-methoxycarbonyl-propyl) -carbamoylethyl] -carbamoylmethyl-D-glucopyranose as hydrate, m.p. 80-90 ° C, [α] g ° = + 25 ° (CH cpH, c = 1.017), Rf value = 0.23 in CHCl 3: ethanol = 9: 1 (silica gel, thin layer plate, Merck).

Example 20

Analogously to Example 16, ethyl 2-benzamido-2-des-oxy-3-O-carboxymethyl- / 3-D-glucopyranoside ethyl-2-benzamido-2-deoxy-3-O- [L-1- (D-1,3-bis-methoxycarbonyl-propyl) -carbamoyl-ethyl] -carbamoyl-methyl- (3-D-glucopyranoside, m.p.

127-135 ° C, [α] g ° = -17 ° (CH $> H, c = 1.024), Rf = 0.26 in ethyl acetate: acetone = 2: 1 (silica gel, thin layer plates,

Merck).

Example 21

Analogously to Example 12, 2-benzamido-2-deoxy-3-O- [L-1- (D-1-N-benzyl-carbamoyl-3-carboxypropyl) -carbamoyl-ethyl-carbamoylmethyl-D-glucopyranose is obtained.

In this case, the γ-carboxy group in the glutamic acid is released from a tert-butyl ester by hydrolysis with "Dowex-50" ®H + itetrahydrofuran water.

Example 22

Analogously to Example 12, 2-benzamido-2-deoxy-3-O- [L-1- (D-1-N-carbamoylmethyl-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl-D-glucopyranose is obtained, m.p.163-170 ° C. The starting material for the peptide moiety is N-tert-butoxycarbonyl-L-alanyl-D-glutamic acid-7-benzyl ester α-glycine amide.

Example 23

Analogously to Example 12, the hydrochloride of La-amino-valeroyl-D-glutamic acid "o:" amide-7-tert-butyl ester 2-benzamideO "2-deoxy-3" O- [L1- (D1-carbamoyl-3) is obtained. -carboxyprop-yl) -carbamoylbutyl] -carbamoylmethyl-D-glucopyranose.

Example 24

Analogously to Example 12, 2-benzamido-2-deoxy-3-O- [L-1- (D-1-carbamoyl-3-carboxypropyl) -carbamoylpropyl] -carbamoylmethyl-D-glucopyranose is obtained.

Example 25

Analogously to Example 12, 2-benzamido-2-deoxy-3-O- [L1- (D1-carbamoyl-3-carboxypropyl) -carbamoyl-2-methyl-propyl] -1-carbamoylmethyl-D-glucopyranose, [α ] δ® = + 32 ° (water, c = 0.78).

Example 26

A solution of 10.7 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O- [(D1-carbamoyl-3-carboxypropyl) -carbamoylmethyl-carb-amoylmethyl] -2-deoxy-α-D-glucopyranoside -benzyl ester in 200ml glacial acetic acid and 100ml water are hydrogenated with 2g of 5% spalladium coal at normal pressure and at room temperature for 57 hours. The catalyst is filtered off, washed with fresh water and the filtrate is evaporated. The residue is taken up in water and filtered over 100 ml of the ion exchanger "Amberlite IR" ® 120 (H + form) and the filtrate is freeze-dried. 2-Acetamido-3-O- [(D-1-carbamoyl-3-carboxypropyl) -carbamoylmethyl-carbamoyl-methyl] -2-deoxy-D-glucose is crystallized from methanol / ethyl acetate and dried in high vacuum. The product containing 1/4 mol of ethyl acetate shows the optical rotation [a] § ° = + 27 ° ± 1 ° (water, c = 0.944).

The starting material used can be prepared as follows: 8 g of N-tert-butoxycarbonyl-glycyl-D-isoglutamiribenzyl ester are dissolved at room temperature and excluding moisture in a mixture of 18 ml of 1,2-dichloroethane and 8.4 ml of trifluoroacetic acid and left in 16 hours. The reaction mixture is diluted with 200 ml of tetrahydrofuran and neutralized externally with triethylamine, and a solution of 8.3 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O-carboxy-methyl-2-deoxy-α-D-glucopyranoside is added. in 100 ml of tetrahydrofuran and 2.52 ml of triethylamine. After the addition of 5.05g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ), stir for 24 hours at room temperature. The crystallized product is extracted, washed with tetrahydrofuran and ether and dried. The obtained benzyl-2-acetamido-4,6-O-benzylidene-3-O- [(D1-carbamoyl-3-carboxy-propyl) -carbamoylmethyl-carbamoylmethyl] -2-deoxy-α-D-glucopyranoside benzyl ester shows the optical rotation [α 8 ° - + 66 ° ± 1 ° (Ν, Ν-dimethylformamide, c = 1.308).

Example 27

A solution of 4.5 g of methyl 2-acetamido-3-O - [[1- (D-1-carbamoyl-3-carboxypropyl) -carbamoyl-ethyl] -carbamoylmethyl) -2-deoxy-αD -glucopyranoside benzyl ester in 125 ml of 50% sulfuric methanol is hydrogenated with 1.0 g of 5% palladium hydrocarbon at normal pressure and room temperature to take up 178 ml of hydrogen. The catalyst is filtered off and the evaporator filtrate in water jet vacuum. The residue is dissolved in 50 ml of distilled water and freeze dried. There is thus obtained methyl 2-acetamido-3-O - [[1- (D1-carbamoyl-3-carboxypropyl) -carbamoyl-ethyl] -carbamoymethyl) -2-deoxy-α-D-glucopyranoside with [α] §0 = + 49 ± 1 ° (water, c = 0.939).

The starting material can be prepared as follows: 8.1 g of N-tert-butoxycarbonyl-L-alanyl-D-isoglutamine-benzyl ester is dissolved at room temperature and excluding moisture in a mixture of 8.1 ml of 1,2-dichloroethane and 8, 1 ml trifluoroacetic acid and leave for 16 hours. The reaction mixture is diluted with 200 ml of tetrahydrofuran, neutralized under exterior cooling with triethylamine, and a solution of 7.62 g of methyl 2-acetamido-4,6-O-benzylidene-3-O-carboxymethyl-2-deoxy-α-D-glucopyranoside is added. and 2.77 ml of triethylamine in 100 ml of tetrahydrofuran. After adding 5.0 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ), the reaction mixture is heated to 40 ° C, stirred at this temperature for 30 hours and allowed to stand for additional 24 hours at room temperature. The precipitated product, methyl 2-acetamido-4,6-O-benzylidene-3-O - <[L-1- (D-1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl) -2-deoxy -or-D-glucopyranoside benzyl ester, extracted, washed with tetrahydrofuran and ether and dried, [a.] 2 ° - + 58 ° ± 1 ° (N, N-dimethylformamide, c = 1.125).

A solution of 10.5 g of methyl 2-acetamido-4,6-O-benzylidene-3-O - [[1- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-aD -glucopyranoside benzyl ester in 320 ml of glacial acetic acid is diluted with 200 ml of water and the whole is stirred for 2 hours at 50-55 ° C. After cooling, the solution is evaporated to dryness, to the residue is added four more times 100 ml of water and evaporated each time. There is thus obtained methyl 2-acetamido-3-O - <[L- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-α-D-glucopyranoside benzyl ester with [<x] § 0 = + 64 ° ± 1 ° (Ν, Ν-dimethylformamide, c = 1.268).

Example 28

A solution of 3.0 g of methyl 2-acetamido-3-O - [[1- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-6-o-stearoyl-aD -glucopyranoside-benzylesteii 100 ml of ethanol and 100 ml of tetrahydrofuran are hydrogenated with 0.6 g of 5% palladium carbon at room temperature and normal pressure, filtered from the catalyst and evaporated in water jet vacuum to dryness. The crystalline residue is methyl 2-acetamido-3-O - <[L- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl} -2-deoxy-6-o-stearoyl-α-D-glucopyranoside with [α] δ + = + 50 ° ± 1 ° (N, N-dime thyl formamide, c = 0.921).

The starting material can be prepared as follows:

To a solution of 3.98 g of methyl 2-acetamido-3-O - [[1- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoyl-methyl) -2-deoxy-o: -D-glucopyranoside Benzyl ester in 40 ml of absolute pyridine is added dropwise over a period of 3 hours with a solution of 2.12 g of stearic acid chloride in 20 ml of 1,2-dichloroethane and stirring for 18 hours at room temperature with stirring and exclusion of moisture at 0-5 ° C. The reaction mixture is diluted with chloroform, washed with water, ice cold 2N hydrochloric acid and water, dried over magnesium sulfate and evaporated to dryness. The product, methyl 2-acetamido-3-O - [[1- (D1-carbamoyl-3-carboxypropyl-yl) -carbamoylethyl] -carbamoylmethyl) -2-deoxy-6-O-stearoyl-α-D-glucopyranoside benzyl ester, crystallized from ethanol / ether, [α] fP = + 22 ° ± 1 ° (chloroform, c = 1.030).

Example 29

A solution of 6.8 g of benzyl-2-acetamido-3-O - <[L-1- (D-1,3-dicarbamoylpropyl) -carbamoylethyl] -carbamoylmethyl> ~ 2-deoxy-α-D-glucopyranoside in 200 ml % aqueous methanol - hydrogenated with 5% palladium coal 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. There is thus obtained 2-acetamido-3-O - <[L-1- (D-1,3-dicarbamoylpropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-D-glucose, which contains 1.24 moles of water which a white powder with [or] §0 = + 7 ° ± 1 ° (water, c = 0.514).

The starting material can be prepared as follows:

To 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 allowed to stand for 48 hours at room temperature. After distillation of the solvent, 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- (D1,3-dicarbamoylpropyl) -carbamoylethyl] -carbamoylmethyl) -2-deoxy-o: - D-glucopyranoside in 120 ml of glacial acetic acid is diluted with 80 ml of water and stirred for 3 hours at 60 ° C. Then the reaction mixture is cooled, evaporated and the residue is added three more times 100 ml of water, each of which is distilled off. The obtained benzyl-2-acetamido-3-O - [[L-1- (D-1,3-dicarbamoylpropyl) -carbamoylethyl] -carbamoyl-methyl) -2-deoxy-α-D-glucopyranoside is recrystallized from methanol, m.p. 223-225 ° C.

Example 30

A solution of 5.7 g of benzyl-2-acetamido-3-O- <D1- [(D1-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 coal at normal pressure and room temperature. The catalyst is filtered off and the filtrate is evaporated. The residual 2-acetamido-3-O- <D1- [(D1-carbamoyl-3-carboxypropyl) -carbamoylmethyl-J-carbamoylpropyl) -2-deoxy-D-glucose is taken up in 50ml of water and lyophilized, [α = +46 ° ± 1 ° (water, c = 0.630).

The starting material used can be prepared as follows: 5.1 g of N-tert-butoxycarbonyl-glycyl-D-isoglutamine benzyl ester is dissolved at room temperature and excludes humidity in a mixture of 5.1 ml of 1,2-dichloroethane and 5, 1 ml trifluoroacetic acid and leave for 16 hours. This solution is diluted with 100 ml of tetrahydrofuran, neutralized under exterior cooling with triethylamine and a single solution of 6.3 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O- (D1-carboxypropyl) -2-deoxy-aD is added. -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, evaporate to dryness and the residue is partitioned between ethyl acetate and water. The organic phase is also washed with medical cold 2N hydrochloric acid, water, a saturated sodium hydrogen carbonate solution and water and evaporated. The residual benzyl-2-acetamido-4,6-O-benzylidene-3-O- <D1- [(D1-carb-amoyl-3-carboxypropyl) -carbamoylmethyl-carbamoylpropyl> -2-deoxy-α-D-glucopyranoside benzyl ester is crystallized fratetrahydrofuran / ether, [α = + 76 ° ± 1 ° (N, N-dimethylformamide, c = 0.457).

A mild acidic hydrolysis of this product in 60% aqueous acetic acid leads to benzyl-2-acetamido-3-O- <D1- [(D1-carbamoyl-3-carboxypropyl) -carbamoylmethyl] -carbamoyl-propyl> -2-deoxy -aD-glucopyranoside benzyl ester crystallizing from methanol / ether, m.p. 180-185 ° C / [α] D = + 87 ° ± 1 ° (methanol, c = 1.035).

Example 31 4 g of N- tert -butoxycarbonyl-L-alanyl-D-isoglutamine benzyl ester are dissolved at room temperature and with the exception of humidity in a mixture of 4 ml of trifluoroacetic acid and 4 ml of 1,2-dichloroethane and leave for 16 hours. This reaction mixture is now diluted with 30 ml of 1,2-dichloroethane and neutralized under exterior cooling with triethylamine, and a solution of 3.7 g of benzyl-2-acetamido-3-O-carboxy-methyl-2-deoxy-α-D-glucopyranoside and 1 g , 38 ml triethylamine i100 ml tetrahydrofuran. After adding 2.6 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) charger mixture, stand for 24 hours at room temperature and evaporate 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. Thus, benzyl-2-acetamido-3-O - <[L-1- (D-1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-de-oxy-α-D-glucopyranoside benzyl ester is recrystallized from ethanol, m.p. 208-212 ° C, [α] 25 D = + 77 ° ± 1 ° (N, N-dimethylformamide, c = 0.546).

After the hydrogenolytic cleavage of the two benzyl groups as described in Example 2, 2-acetamido-3-O - <[L- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2 is obtained. deoxy-D-glucose.

Example 32

A solution of 3.8 g of benzyl-2-acetamido-3-O - [[1- (D-1,3-dicarboxypropyl) -carbamoylethyl] -carbamoylmethyl) -2-deoxy-or-D-glucopyranoside dimethyl ester in 100 ml of methanol is hydrogenated in the presence of 5% palladium coal at normal pressure and room temperature. After the hydrogen uptake is complete, the catalyst is filtered off and the filtrate is evaporated to dryness. The residue is taken up in 70 ml of water and lyophilized. The foam obtained is 2-acetamido-3-O - <[L- (D1,3-dicarboxy-propyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-D-glucose-dimethyl ester with [ot] §0 = + 23 ° ± 1 ° (water, c = 0.814).

The starting material used can be prepared as follows:

To a solution of 12.9 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O-carboxymethyl-2-deoxy-1 H -D-glucopyranoside and 4.0 ml of triethylamine in 100 ml of Ν, Ν- dimethylformamide is added 8.1 g of L-alanyl-D-glutamic acid dimethyl ester hydrochloride and 6.95 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ), and the mixture is allowed to stand at room temperature for 20 hours, then the solvent is evaporated off. the residue in chloroform and washing this solution with water, ice-cold 2 N hydrochloric acid, water, a saturated sodium hydrogen carbonate solution and water. After drying with magnesium sulfate, evaporation is evaporated. The residue is extracted with hot ethanol and the dissolved benzyl-2-acetamido-4,6-O-benzylidene-3-O - <[L-1- (D-1,3-dicarboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2- de-oxy-αD-glucopyranoside dimethyl ester is filtered off and dried, [a] §0 = + 22 ° ± 1 ° (chloroform, c = 1.160).

A solution of 15.6 g of benzyl-2-acetamido-4,6-O-benzylidene-3- O - <[L-1- (D-1,3-dicarboxypropyl) -carbamoylethyl] -carbamoylmethyl) -2-deoxy -o; -D-glucopyranoside dimethyl ester in 420 ml glacial acetic acid and 280 ml water is heated to 10 ° C. After 4 hours of stirring at this temperature, the reaction mixture is cooled and evaporated to dryness. To the residue is added three times 100 ml of water and evaporated each time. The residue is also taken up in chloroform. This solution is washed with water, dried with magnesium sulfate and evaporated to dryness. There is thus obtained benzyl-2-acetamido-3-O - <[L- (D1,3-dicarboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-α-D-glucopyranoside dimethyl ester as an angular resin with [α = + 31 ° ± 1 ° (chloroform, c = 1.070).

Example 33

A solution of 6.1 g of benzyl-3-O - [[1- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-2-propionamido-α-D-glucopyranoside benzyl ester in 200 ml of tetrahydrofuran Water 2: 1 is hydrogenated under the presence of 0.6 g of 5% palladium coal at normal pressure and room temperature. Upon completion of hydrogen uptake, the catalyst is suctioned off and the filtrate is evaporated. To the residue is added 150 ml of water and it is hydrogenated in the presence of 5% palladium coal until no more hydrogen is taken up. The catalyst is filtered off and lyophilized. There is thus obtained 3-O - <[L-1- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoyl-methyl) -2-deoxy-2-propionamido-D-glucose with [α] §0 = + 8 ° (water, c = 1.146).

The starting material used can be prepared as follows:

To a solution of 90 g of benzyl-2-acetamido-2-deoxy-o: -D-glucopyranoside in 900 ml of Ν, Ν-dimethylformamide is added sub-stirring, exclusion of moisture and ice-water cooling 0.3 ml of methanesulfonic acid. Then, in a one hour solution of 60 ml of isopropenylmethyl ether in 240 ml of N, N-dimethylformamide is added dropwise, stirred for another two hours at room temperature and adjusted with triethylamine for basic reaction. The solvent is distilled off and the residue is taken up in ethyl acetate. This solution is washed with water, dried over magnesium sulfate and evaporated to dryness. The product, benzyl-2-acetamido-2-deoxy-4,6-O-isopropylidene-α-D-glucopyranoside, is crystallized from ether, m.p.136-137 ° C, [α] g ° = + 103 ° ± 1 ° (chloroform, c = 1.125).

Dissolve 52.5 g of benzyl-2-acetamido-2-deoxy-4,6-O-isopropylidene-α-D-glucopyranoside in a solution of 225 g of potassium hydroxide in 750 ml of ethanol and 40 ml of distilled water and boil for 4 1/2 hours. under reflux. After cooling, the reaction mixture is evaporated to half volume and poured into ice. Extract with chloroform, wash the organic phase with water, dry with magnesium sulfate and evaporate dryness. The residue, benzyl-2-amino-2-deoxy-4,6-O-isopropylidene-α-D-glucopyranoside, is crystallized from ether, mp 145-146 ° C, [o:] §0 = + 117 ° ± 1 ° (chloroform, c = 1.295).

To 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 added a solution of 16.0 g of potassium hydrogen carbonate in 192 ml of distilled water and cooled to 0 ° C. Now, while stirring, 8.16 g of propionic acid chloride is added dropwise over 20 minutes and the mixture is stirred at this temperature for a further 50 minutes. Now the organic phase is separated, washed with water, dried with magnesium sulfate and evaporated to dryness. The product, benzyl-2-deoxy-4,6-O-isopropylidene-2-propionamido-α-D-glucopyranoside; crystallize from ethyl acetate / petroleum ether, m.p. 121-122 ° C, [α] 25 D = + 112 ° ± 1 ° (choroform, c = 0.977).

To 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 added 1.25 g of sodium hydride (Techn. Fluka) and stirred for 2 hours. hours at 40 ° C. It is then cooled to -5 to -10 ° C and 4.2 ml of bromoacetic acid ethyl ester are added. After another 20 minutes, 10 ml of ethanol is 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 is washed with water, dried with magnesium sulfate and evaporated. The product, benzyl-3-O-carboxymethyl-2-deoxy-4, 6-O-iso-propylidene-2-propionamido-α-D-glucopyranoside ethyl ester, crystallized from ether / petroleum ether, m.p. 94-95 ° C, [a] §0 = + 145 ° ± 1 ° (chloroform, c = 1.218).

To 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 added 22.5 ml of 1N sodium hydride solution. After ester hydrolysis, 7.5 ml of 1N hydrochloric acid is added and evaporated to dryness. The product is dissolved in 50 ml of N, N-dimethylformamide and condensed with 15 mmol of L-alanine-D-isoglutamine benzyl ester trifluoroacetate in the presence of 3.72 g of 2- ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ). Then, evaporate to dryness and take up the residue in chloroform. This solution is washed with water, ice cold 2 N hydrochloric acid, water, a saturated sodium bicarbonate solution and water, dried with magnesium sulfate and evaporated to dryness. The product is crystallized from diluted ethanol, mp 177-180 ° C, [α] g ° = + 71 ° ± 1 ° (chloroform, c = 1.047).

To a solution 8.1 g of benzyl-3-O - <[L- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-4,6-O-isopropylidene-2-propionamido-o : -D-glucopyranoside benzyl ester in 150 ml of methanol is added 15 ml of 1 N hydrochloric acid and the mixture is allowed to stand for one hour at room temperature. Now add 15 ml of 1 N sodium hydroxide solution and evaporator to dryness. The obtained benzyl 3-O - <[L-1- (D-1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-2-propionamido-α-D-glucopyranoside benzyl ester is crystallized from methanol / water and dried. Mp. 208-210 ° C = + 75 ° ± 1 ° (N, N-dimethylformamide, c = 1,120).

Example 34

By catalytic hydrogenation of benzyl-3-O - <[L1- (D1-carb-amoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-caprinoylamido-2-deoxy-D-glucopyranoside benzyl ester in Ν -dimethylformamide in the presence of 5% spalladium coal, 3-0 - <[L1- (D1-carb-amoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-caprinoylamido-2-deoxy-aD-glucose is obtained as white foam with [or = + 11 ° (water, c = 1.052).

The starting material used can be prepared as follows:

Analogously to Example 46, 24.7 g of benzyl-2-amino-2-deoxy-4,6-O-isopropylidene-or-D-glucopyranoside are reacted with 16.7 g of caprinoyl choride and worked up. Benzyl-2-caprinoylamido-2-deoxy-4,6-O-isopropylidene-or-D-glucopyranoside is obtained [o; = + 81 ° ± 1 ° (chloroform, c = 1.019).

To a solution of 3.8 g of benzyl-2-caprinoylamido-2-deoxy-4,6-O-isopropylidene-or-D-glucopyranoside in 40 ml of acetonitrile 0.4 g of technical grade sodium hydride is added and stirred for 2 hours. 40 ° C. Then, this mixture is cooled to -10 ° C, 1.4 ml of bromoacetic acid ethyl ester is added and stirred for another hour at 0 ° C. Worked up as Example 33, benzyl-3-O-carboxymethyl-2-caprinoylamido-4,6-O-isopropylidene-2-deoxy-or-D-glucopyranoside ethyl ester is obtained as a tan oil with [or] g = + 114 ° ± 1 ° (chloroform, c = 1.224).

Analogously to Example 33, the ethyl ester is hydrolyzed, the product is condensed with L-alanine-D-isoglutamine benzyl ester trifluoroacetate in the presence of EEDQ, and the isopropylidene ketal is cleaved by mild acidic hydrolysis. Thus, benzyl 3-O - [[L- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl) -2-caprinoylamido-2-deoxy-or-D-glucopyranoside benzyl ester is obtained as a yellow resin, snip. 192-193 ° C.

Example 35

A 5% solution of benzyl-2-acetamido-2-deoxy-3-O - <[L1- (D1,3-bis-methylcarbamoylpropyl) -carbamoylethyl] -carbamoylmethyl> -AD-glucopyranoside in distilled water methanol 1: 1 is hydrogenated with 5% palladium carbon. The catalyst is filtered off and the filtrate is evaporated. The product, 2-acetamido-2-deoxy-3- [0 - [[1- (D1,3-bis-methyl-carbamoylpropyl) -carbamoylmethyl] -carbamoylmethyl> -D-glucose, is freeze-dried, [a] = + 31 ° (water, c = 0.41).

The starting material can be prepared as follows:

To 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 Ν-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 allowed to stand for 48 hours at room temperature. The solvent is now distilled off, and to the oily residue, the insoluble filter is filtered off, washed with water and dried. This product is stirred twice more with ether, filtered off and dried and is benzyl-2-acetamido-4,6-O-benzylidene-2-deoxy-3-O - <[L1- (D1,3-bis-methylcarbonic acid). amoylpropyl) -carbamoylethyl] -carbamoylmethyl> -αD-glucopyranoside.

By mild acid hydrolysis, as described in Example 29, with 60% acetic acid, the benzylidene group is decomposed and the product, benzyl-2-acetamido-2-deoxy-3-O - <[L- (D1,3-bis-methylcarbamoylpropyl) ) -carbamoylethyl] -carbamoylmethyl> -αD-glucopyranoside, is isolated.

Example 36

A 5% aqueous solution of benzyl-2-acetamido-3-O - [[1- (D1-carbamoyl-3-carboxypropyl) -carbamoyl-2-hydroxyethyl] -carbamoylmethyl} -2-deoxy-α-D-glucopyranoside is hydrogenated in the presence of 5% palladium coal, filter and freeze-dry. There is thus obtained 2-acetamido-3-O - [[1- (D1-carbamoyl-3-carboxypropyl) -carbamoyl-2-hydroxyethyl] -carbamoylmethyl) -2-deoxy-D-glucose with [α 10 ° (water, c- 1.653).

The starting material can be prepared as follows:

To a solution of 3.7 g of benzyl-2-acetamido-3-O-carboxy-methyl-2-deoxy-G-D-glucopyranoside and 1.38 ml of triethylamine in 150 ml of tetrahydrofuran is added 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), and the mixture is left for 20 hours at room temperature. 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 solvent free. There is thus obtained benzyl-2-acetamido-3-O - [[1- (D1-carbamoyl-3-carboxylpropyl) -carbamoyl-2-hydroxyethyl] -carbamoylmethyl> -2-deoxy-or-D-glucopyranoside tert-butyl ester.

By mild acid hydrolysis, as described in Example 11, the tert-butyl ester is hydrolyzed and benzyl-2-acetamido-3-O - <[L- (D1-carbamoyl-3-carboxypropyl) -carbamoyl-2-hydroxyethyl is obtained. ] carbamoylmethyl> -2-deoxy-alpha-D-glucopyranoside.

Example 37

A 5% solution of benzyl-2-acetamido-3-O - [[1- (D1-carb-amoyl-3-carboxypropyl) -carbamoylbutyl] -carbamoylmethyl> -2-deoxy-α-D-glucopyranoside in methanol / water 1: 1 is hydrogenated under the presence of 5% palladium coal, filtered and evaporated. The residue is dissolved in distilled water and freeze dried. There is thus obtained 2-acetamido-3-O - [[L-1- (D-1-carbamoyl-3-carboxypropyl) -carbamoylbutyl] -carbamoylmethyl-2-deoxy-D-glucose, [a] §0 = + 12 ° (water, c = 0.722).

The starting material can be prepared as follows:

To a solution of 3.7 g of benzyl-2-acetamido-3-O-carboxy-methyl-2-deoxy-G-D-glucopyranoside and 1.38 ml of triethylamine in 100 ml of tetrahydrofuran is added 3.28 g of L-norvaline. D-isoglutamine-tert-butyl ester hydrochloride and 2.6 g of 2-ethoxy-N-ethoxy-carbonyl-1,2-dihydroquinoline (EEDQ), and the mixture is left at room temperature for 24 hours. The reaction mixture is worked up as in Example 36. Benzyl-2-acetamido-3-O - <[L-1- (D1-carbamoyl-3-carboxypropyl) -carbamoylbutyl] -carbamoylmethyl> -2-deoxy-o 1-D-glucopyranoside tert-butyl ester, m.p.

202-204 ° C.

The tert-butyl ester is cleaved by mild acidic hydrolysis.

Example 38

A solution of 8.3 g of benzyl-3-O - [[1- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-2-stearoylamido-α-D-glucopyranoside benzyl ester in 200 ml glacial acetic acid hydrogenated in the presence of 5% spalladium coal at normal pressure and room temperature. After reprocessing, 3-O - <[L1- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-2-stearoylamido-D-glucose is obtained as a white powder, [a] β0 = +24 ° (dimethylformamide, c = 0.663).

The starting material used can be prepared as follows:

To a solution of 6.2 g of benzyl-4,6-O-benzylidene-2-deoxy-2-stearoylamido-α-D-glucopyranoside in 150 ml of N, N-d'imethylformamide is added 0.75 g of sodium hydride (technical) and stir for 1 1/2 hours at 40 ° C. Now, the mixture is cooled to -5 ° C, 2.2 ml of bromoacetic acid ethyl ester is added and stirred for 4 hours at room temperature. After adding 10ml of glacial acetic acid and 400ml of distilled water, the chloroform is extracted, the organic phase is washed with water, dried and evaporated to dryness. The obtained benzyl 4,6-O-benzylidene-3- O-carboxymethyl-2-deoxy-2-stearoylamido-α-D-glucopyranoic acid ethyl ester is crystallized from ethanol, m.p. 151-154 ° C, [α] §0 = + 94 ° ± 1 ° (chloroform, c = 1.186).

To a solution of 23.4 g of benzyl-4,6-O-benzylidene-3-O-carboxymethyl-2-deoxy-2-stearoylamido-α-D-glucopyranoside ethyl ester in 334 ml of methanol and 334 ml of tetrahydrofuran is added 50.1 ml of 1 N sodium hydroxide solution, which is stirred for 90 minutes at 50 ° C and evaporated. The residue is stirred in ice water and the product is suctioned off, washed with water, dried and recrystallized from ethanol. Benzyl 4,6-O-benzylidene-3-O-carboxymethyl-2-deoxy-2-stearoylamido-ol-D-glucopyranoside sodium salt.1H 2 O melts at 225-242 ° C (with decomposition), [α ] §0 = + 45 ° ± 1 ° (chloroform, c = 1.097).

To a solution of 14.4 g (20 mmol) of benzyl-4,6-O-benzylidene-3-O-carboxymethyl-2-deoxy-2-stearoylamido-α-O-gluco-pyranoside sodium salt monohydrate in 120 ml tetrahydrofuran is added 20 mmol of L-alanine-D-isoglutamine benzyl ester trifluoroacetate and 4.95 g (20 mmol) of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ). The mixture is left at room temperature for 20 hours and evaporated to dryness. The residue is dissolved 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. The product, benzyl-4,6-O-benzylidene-3-O - <[1- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-2-stearoylamido-α-D-glucopyranoside benzyl ester, recrystallized from ethanol, [α] 20 = + 62 · 1 1 · (Ν, Ν-dimethylformamide, c = 1.148).

Mild acid hydrolysis of this benzylidene derivative in 65% acetic acid leads to benzyl 3-0 - <[L- (D1-carbamoyl-3-carb-oxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-2-stearoylamido-aD -glucopyranoside benzyl ester, m.p.

188-189 ° C = + 59 ° ± 1 ° (Ν, Ν-dimethylformamide, c = 1.022).

Example 39

A 5% aqueous solution of benzyl-2-acetamido-3-O - [[1- (D1-carbamoyl-3-carboxypropyl) -carbamoyl-2-methylpropyl] -carbamoylmethyl> -2-deoxy-αD-glucopyranoside is hydrogenated in the presence of 5% palladium carbon, filtered and the filtrate lyophilized. There is thus obtained 2-acetamido-3-O- [L1- (D1-carbamoyl-3-carboxypropyl) -carbamoyl-2-methylpropyl] -carbamoylmethyl-2-deoxy-D-glucose, [a W - + 11 ° (water , c = 1.096).

The starting material can be prepared as follows:

To a solution of 3.7 g of benzyl-2-acetamido-3-O-carboxy-methyl-2-deoxy-α-D-glucopyranoside and 1.38 ml of triethylamine in 100 ml of tetrahydrofuran is added 3.38 g of L-valine-D-isoglutamine. -tert-butyl ester hydrochloride and 2.6 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline. The mixture is left at room temperature for 24 hours and worked up as described in Example 38. Thus, benzyl-2-acetamido-3-O - <[L- (D1-carbamoyl-3-carboxypropyl) -carbamoyl-2-methylpropyl] - carbamoyl-methyl) -2-deoxy-or-D-glucopyranoside tert-butyl ester, - dehydrolyses mildly acid to benzyl-2-acetamido-3-O - [[1- (D1-carbamoyl-3-carboxypropyl) -carbamoyl 2-methylpropyl] -carbamoylmethyl> -2-deoxy-α-D-glucopyranoside.

Example 40

A 5% solution of benzyl-2-acetamido-3-O - [[1- (D1-carb-amoyl-3-carboxypropyl) -carbamoyl-3-methylbutyl] -carbamoyl-methyl) -2-deoxy aD-glucopyranoside in 50% aqueous methanol is hydrogenated in the presence of 5% palladium coal, filtered and evaporated. The residue is dissolved in distilled water, filtered again and freeze dried. There is thus obtained 2-acetamido-3-O - <[L- (D1-carbamoyl-3-carboxypropyl) -carbamoyl-3-methylbutyl] -carbamoylmethyl> -2-deoxy-D-glucose, [a] §0 = + 16 ° (water, c = 1.06).

The starting material used can be prepared as follows:

To a solution of 3.7 g of benzyl-2-acetamido-3-O-carboxy-methyl-2-deoxy-α-D-glucopyranoside and 1.58 ml of triethylamine in 100 ml of tetrahydrofuran is added 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). The mixture is left at room temperature for 24 hours and worked up as described in Example 38. Benzyl-2-acetamido-3-O - <[L-1- (D-1-carbamoyl-3-carboxypropyl) -carbamoyl-3-methylbutyl] - carbamoylmethyl> -2-deoxy-α-D-glucopyranoside tert-butyl ester, thereby causing mild acid hydrolysis to benzyl-2-acetamido-3-O - <[L1- (D1-carbamoy1-3-carboxypropyl) -carbamoy1-3 -methyl-butyl] -carbamoylmethyl) -2-deoxy-1 H -D-glucopyranoside.

Example 41

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

The starting material can be prepared as follows:

To a solution of 3.7 g of benzyl-2-acetamido-3-O-carboxy-methyl-2-deoxy-aDg, lucopyranoside and 1.38 ml of triethylamine in 100 ml of tetrahydrofuran is added 3.24 g of NaL-aminobutyryl-D-isoglutamine. -tert-butyl ester hydrochloride and 2.6 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline, and after 24 hours at room temperature the mixture is worked up as described in Example 36. The resulting benzyl-2-acetamido-3-O- [L- (D1-carbamoyl-3-carboxypropyl) -carbamoylpropyl] -carbamoylmethyl> -2-deoxy-α-D-glucopyranoside tert-butyl ester is then hydrolyzed mildly to benzyl-2-acetamido-3-O- [1- (D1-carbamoyl-3-carboxypropyl) -carbamoylpropyl] -carbamoylmethyl) -2-deoxy-αD-glucopyranoside.

Example 42

A 5% solution of benzyl-2-acetamido-3-O - [[L-1- (D-1-carbamoyl-3-carboxypropyl) -carbamoylphenylmethyl] -carbamoylmethyl) -2-deoxy-α-D Glucopyranoside in 50% aqueous methanol is hydrogenated in the presence of 5% spalladium coal at normal pressure and room temperature, filtered from the catalyst and evaporated. The residue is dissolved in distilled water and lyophilized and is 2-acetamido-3-O - <[L-1- (D-1-carbamoyl-3-carboxypropyl) -carbamoylphenylmethyl] -carbamoylmethyl> -2-deoxy-D-glucose.

The starting material can be prepared as follows:

To a solution of 3.7 g of benzyl-2-acetamido-3-O-carboxy-methyl-2-deoxy-α-D-glucopyranoside and 1.38 ml of triethylamine in 100 ml of tetrahydrofuran is added 3.72 g of 1-phenylglycine D-iso -glutamine-tert-butyl ester hydrochloride and 2.6 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinol ± n (EEDQ). The mixture is left at room temperature for 24 hours and worked up as described in Example 36. There is thus obtained benzyl-2-acetamido-3-O - <[L- (D1-carbamoyl-3-carboxypropyl) -carbamoylphenylmethyl] -carbamoylmethyl> 2-Deoxy-α-D-glucopyranaside tert-butyl ester, which is mildly hydrolyzed under acidic conditions to benzyl-2-acetamido-3-O - <[L- (D1-carbamoyl-3-carboxy-propyl) -carbamoylphenylmethyl] -carbamoylmethyl) -2-deoxy-alpha-D-glucopyranoside.

Example 43

To a solution of 3.5 g of benzyl-2-acetamido-3-O - <[L-1- (D-1,3-bis-carbamoylpropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-4,6- O-isopropylidene-3-D-glucopyranoside in 50 ml of vinegar is added dropwise at 55 ° C dropwise over 15 minutes 50 ml of distilled water and stirred for one hour at this temperature. After cooling, another 100 ml of distilled water is added and the solvent evaporated in vacuo is evaporated. Add 20 ml of water to the residue and freeze-dry. There is thus obtained benzyl-2-acetamido-3-O - [[1- (D1,3-bis-carbamoylpropyl) -carbamoylethyl] -carbamoylmethyl-2-deoxy-0-D-glucopyranoside with [a] §0 = -44 ° ± 1 ° (N, N-dimethylformamide, c = 0.989).

The starting material used can be prepared as follows:

To a solution of 30 g of benzyl-2-acetamido-2-deoxy- / 3-D-glucopyranoside in 300 ml of Ν, Ν-dimethylformamide is added sub-stirring, external cooling and moisture exclusion 0.1 ml of methanesulfonic acid. Now, in one hour, a solution of 20 ml of isopropenylmethyl ether in 60 ml of Ν-dimethylformamide is added dropwise over a period of two hours, stirred at room temperature and adjusted with triethylamine for basic reaction. After evaporation of the solvent, the benzyl-2-acetamido-2-deoxy-4,6-O-isopropylidene-β-D-glucopyranoside is crystallized from ethyl acetate, m.p. 194 ° C, [α] 2 ° = -104 ° ± 1 ° (chloroform, c = 0.850).

To a solution of 36.2 g of benzyl-2-acetamido-2-deoxy-4,6-O-isopropylidene-β-D-glucopyranoside in 400 ml of acetonitrile and 100 ml of Ν, dim-dimethylformamide is added 4.95 g of sodium hydride (technically ) and stir for 2 hours at 40 ° C. Now, this mixture is cooled to -10 ° C, 17.2 ml of bromoacetic acid ethyl ester is added and stirred for 30 minutes at 0 ° C. Now, 40 ml of ethanol is added, neutralized with glacial acetic acid and evaporated to dryness. The residue is partitioned between ether and distilled water, the ether phase is washed with water, dried with magnesium sulfate and evaporated. The product, benzyl-2-acetamido-3-O-carboxymethyl-2-deoxy-4,6-O-isopropylidene-β-D-glucopyranoside ethyl ester, is crystallized from ether, mp 93-94 ° C, ] g = -49 ° to 1 ° (CHCl 3 c = 1.001).

To a solution of 6.56 g of benzyl-2-acetamido-3-O-carbamoylmethyl-2-deoxy-4,6-O-isopropylidene / 3-D-glucopyranoside ethyl ester in 70 ml of methanol is added 22.5 ml of 1 N sodium hydroxide solution. After ester hydrolysis, 7.5 ml of 1 N hydrochloric acid is added and evaporated to dryness. The resulting sodium salt is dissolved in 50 ml of Ν, Ν-dimethylformamide and 3.7 g of L-alanyl-D-glutamic acid diamide hydrochloride and 3.72 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) are added. After standing for 24 hours at room temperature, evaporate to dryness, dissolve the residue in water and extract with methylene chloride and twice with n-butanol. The product remaining after distilling off the n-butanol is benzyl-2-acetamido-3-O - <[L- 1- (D-1,3-Bis-carbamoylpropyl) -carbamoylethyl] -carbamoylmethyl-2-de-oxy-4,6-O-isopropylidene- (3-D-glucopyranoside.

Example 44

To a solution 5.1 g of benzyl-2-acetamido-3-O - <[L-1- (D-1,3-bis-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-4.6-0 Isopropylidene-β-D-glucopyranoside dimethyl ester in 120 ml glacial acetic acid is added dropwise at 50 ° C dropwise to 80 ml of distilled water and stirred for 1 hour at this temperature. After evaporation of the acetic acid, benzyl 2-acetamido-3-O - <[L- (D1,3-bis-carboxypropyl) -carbamoylmethyl] -carbamoylmethyl> -2-deoxy- / 3-D-glucopyranoside dimethyl ester is obtained. mp. 195-197 ° C.

The starting material used can be prepared as follows: 6.56 g of benzyl-2-acetamido-3-O-carboxymethyl-2-deoxy-4,6-O-isopropylidene-β-D-glucopyranoside ethyl ester are hydrolyzed as described in Example 43 with sodium hydroxide solution and condensed with 4.24 g of L-alanyl-D-glutamic acid dimethyl ester hydrochloride in the presence of 3.72 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline. After evaporation of the solvent, the residue is taken up with chloroform and washed with water, ice-cold 2N hydrochloric acid, water, a saturated sodium bicarbonate solution and water, dried over magnesium sulfate and evaporated to dryness. Residue Benzyl-2-acetamido-3-O - [[1- (D1,3-bis-carboxypropyl) -carbamoylethyl] -carbamoylmethyl) -2-deoxy-4,6-O-isopropylidene-β-D-glucopyranoside -dimethylester.

Example 45

A 5% solution of benzyl-2-acetamido-3-O - [[1- (D-1-carbamoylmethylcarbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl) -2-deoxy-α-D-glucopyranoside benzyl ester itetrahydrofuran / water 2: 1 is hydrogenated in the presence of 5% palladium coal at normal pressure and room temperature. 50% of the theoretical amount of hydrogen is taken up, the catalyst is filtered off and evaporated. The residue is now dissolved in distilled water and further hydrogenated. After the theoretical amount of hydrogen is taken up, the catalyst is filtered off and the filtrate is freeze-dried. Thus 2-acetamido-3-O - <[L1- (D1-carbamoylmethylcarbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-D-glucose is obtained as a white powder, [a] §0 = + 10 ° (water, c = 0.706).

The starting material can be prepared as follows: 4.8 g of N-tert-butoxycarbonyl-L-alanine-D-glutamine-7-benzyl ester glycinamide monohydrate is dissolved in a mixture of 5 ml of trifluoroacetic acid and 5 ml of 1,2-dichloroethane and left moisture exclusion for 16 hours at room temperature. This solution is diluted with 50 ml of tetrahydrofuran, cooled in an ice bath and neutralized with triethylamine. After adding the solution of 3.7 g of benzyl-2-acetamido-3-carboxymethyl-2-deoxy-α-D-glucopyranoside and 1.38 ml of triethylamine in 100 ml of tetrahydrofuran, 2.6 g of 2-ethoxy-N-ethoxycarbonyl are added to the whole. -1,2-dihydroquinoline and the mixture is left for 24 hours at room temperature and worked up as described in Example 36. Thus benzyl-2-acetamido-3-O - <[L- (D1-carbamoylmethylcarbamoyl-3-carboxypropyl) -carbamoyl] is obtained. ethyl] -carbamoylmethyl> -2-deoxy-α-D-glucopyranoside benzyl ester.

Example 46

A solution of 5.2 g of benzyl-4,6-O-benzylidene-3-O - [[1- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoyl-methyl> -2-deoxy-2-p -tolylsulfonylamino-α-D-glucopyranoside benzyl ester in 120 ml of 70% aqueous acetic acid is hydrogenated under the presence of palladium coal at normal pressure and room temperature. After the theoretical amount of hydrogen is absorbed, the catalyst is filtered off and evaporated to dryness. The residue is dissolved in 30 ml of distilled water and lyophilized, thus obtaining 3-0 - <[L- (D1-carbamoyl-3-carb-oxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-2-p-tolylsulfonylamino -D-glucose as a white powder.

The starting material used can be prepared as follows:

To a solution of 6.0 g of benzyl-2-amino-4,6-O-benzylidene-3-O-carboxymethyl-2-deoxy-α-D-glucopyranoside in 100 ml of methylene chloride and 6.3 ml of triethylamine is added at 5 ° C dropwise. a solution of 3.1 g of p-toluenesulfonic acid chloride in 30 ml of methylene chloride and the mixture is left for 16 hours at room temperature. Then, this solution is washed with water, 2N sodium hydroxide solution, water, cold 2N hydrochloric acid and water, dried over magnesium sulfate and evaporated to dryness. The residue is repeatedly extracted with ether and dried and is benzyl-4,6-O-benzylidene-3-O-carboxymethyl-2-deoxy-2-tolylsulfonylamino-α-D-glucopyranoside, m.p.185-187 ° C, [a] $ 0 = + 75 ° ± 1 ° (chloroform, c = 0.855).

To a solution of 5.7 g of benzyl-4,6-O-benzylidene-3-O-carboxymethyl-2-deoxy-2-tolylsulfonylamino-o: -D-glucopyranoside and 1.01 g of triethylamine in 80 ml Ν , Ν-Dimethylformamide is added 10 mmol of L-alanyl-D-isoglutamine benzyl ester trifluoroacetate and 2.5 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline, and the mixture is allowed to stand for 24 hours at room temperature. to dryness, add water, filter the undissolved, wash with water and ether and dry in vacuo. There is thus obtained benzyl-4,6-O-benzylidene-3-O - [[1- (D1-carbamoyl-3-carboxypropyl) -carbamoylmethyl] -carbamoylmethyl) -2-deoxy-2-tolylsulfonylamino-α -D-glucopyranoside benzyl ester as a white powder.

Example 47 20 g of 2-amino-2-deoxy-3- (carboxymethyl) -D-glucopyranose (30% mixture with KCl) is finely powdered in 100 ml of absolute acetonitrile. With good stirring and light cooling, 26.6 ml of trimethylchiorsilane is first added, then 31.5 ml of triethylamine. After 4 hours at room temperature, in the thin layer chromatogram (silica gel, Merck), the inlet chloroform: methanol = 3: 1 shows practically only the endohydrin positive spot (Rf = 0.7) of the persilylated amino acid. An additional 3.4 ml of triethylamine and 42.5 ml of a 0.62 N solution of 2-methyloxazolinone (5) iabsolut dimethylformamide are added and the mixture is allowed to stand at room temperature for 15 hours. The solution of 2-methyloxazolinone (5) was prepared from 5.85 g of N-acetylglycine and 11.4 g of dicyclohexylcarbodiimide in 40 ml of dimethylformamide, filtering off 85% (9.6 g) of the dicyclohexylurea formed.

After filtration of the salts, partitioning between methylene chloride and water and hydrolysis of the silyl ether in the CH 2 Cl 2 phase with a few drops of acetic acid, 2- (acetaminomethyl-carbonylamino) -2-deoxy-3-carboxymethyl-D-glucopyranose is obtained as a colorless powder by rubbing with ethyl acetate.

The reaction with L-alanyl-D-glutamic acid α-amide-7-benzyl ester trifluoroacetate and 2-ethoxy-N-carbethoxy-1,2-dihydroquinoline in DMF: methylene chloride = 1: 1 occurs analogously to Example 33 After hydrogenation with 5% palladium coal, 2- (acetaminomethylcarbonylamino) -2-deoxy-3-O - <[L- (D1-carbamoyl-3-carboxypropyl) -1-carbamoylethyl] -carbamoylmethyl> - α, β-D-glucose as lyophilisate.

Example 48 3.2 g of 2-amino-2-deoxy-3-carboxymethyl-D-glucopyranose, 2.5 g of K2CO3 and 4.5 g of pivalic acid / 3-nitrophenyl ester are stirred in 100 ml of dimethyl formamide at 100 ° C for 15 hours. The salts are then filtered off, the filtrate is evaporated in oil pump vacuum and poured into an aqueous solution over the "IR-120" ion exchanger. The eluate is shaken with CHCl3 and evaporated in vacuo to a syrup. 2-des-oxy-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) -O 2 -oxazoline in 1000 ml of 2.6 N hydrochloric acid is refluxed for 4 hours. The precipitated debenzoic acid is filtered off and the filtrate is extracted three times with ether. The aqueous phase is adjusted to pH 7 by pH 3 with pH 3 and is treated with activated charcoal. 500 ml and add 3 liters of acetone, cool with ice and suck the precipitate from it. After drying at 40 ° C in vacuo, 247 gives a yellowish powder, of which 85% is KCl and 15% is said compound.

Of the mother liquor, another 22 g of a mixture of the said compound and the associated lactam is obtained.

The reaction of 2-pivaloylamido-2-deoxy-3-carboxymethyl-D-glucopyranose with L-alanyl-D-glutamic acid G; -amide-γ-benzyl-ester trifluoroacetate occurs analogously to Example 33 with 2-ethoxy-N-carbethoxy -1,2-dihydroquinoline in DMF / CH 2 Cl 2 = 1: 1. After hydrogenation with 5% palladium carbon, one obtains 2-trimethylacetamido-2-deoxy-3-O - <[L- (D1-carbamoyl-3-carboxypropyl) -1-carbamoylethyl] -carbamoylmethyl-O-D-glucose as lyophilisate . [α] β0 = -7 ° (methanol, c = 0.6).

Example 49 4.1 g of benzyl-2-acetamido-2-deoxy-4,6-benzylidene-α-D-glucopyranoside is dissolved in 30 ml of Ν, Ν-dimethylformamide and stirred at room temperature up to 40 ° C with 0 5 g of sodium hydride mineral oil dispersion, excluding moisture to form a homogeneous solution. Then, dropwise while cooling at 10 ° C, a solution of 4.76 g of N-iodoacetyl-L-alanyl-D-isoglutamine-Y-benzyl ester in 15 ml of Ν, Ν-dimethylformamide is added and the mixture is stirred at room temperature for 24 hours, then evaporated in vacuo for the syrup.

A solution of 8 g of this compound in 400 ml of 60% acetic acid is heated to 80 ° C for one hour. After cooling, the solution is evaporated to dryness, to the residue is added 2 x 50 ml of water, evaporating to dryness each time. The crystalline residue formed is stirred with a little water, the crystals are aspirated and dried. In this way, manbenzyl-2-acetamido-3-O - [[1- (D1-carbamoyl-3-carboxypropyl-yl) -carbamoylmethyl] -carbamoylmethyl) -2-deoxy-o: -D-glucopyranoside benzyl ester, m.p. 200-202 ° C, [or] 2® = +77 ± 1 °

Cdimethylformamide, c = 0.599).

A solution of 4 g of benzyl-2-acetamido-3-O - [[1- (D1-carb-amoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-α-D-glucopyranoside benzyl ester in 80 ml of methanol is hydrogenated with 0.4 g of 5% palladium coal as catalyst at normal pressure and room temperature until the reaction has ceased. The catalyst is filtered off, washed with a little methanol and the filtrate is evaporated to dryness in water-jet vacuum. The residue is dissolved in 50 ml of distilled water and then hydrogenated with 1 g of 5% palladium-carbon catalyst at normal pressure and room temperature for final reaction. The catalyst is filtered off, washed with a little water and the filtrate is evaporated to dryness. The resulting 2-acetamido-3-O - [[1- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl) -2-deoxy-D-glucose is dried in high vacuum over phosphorus pentoxide, [a] §0 = + 10 ° ± 1 ° (water, c = 0.930).

Example 50

To 5.31 g of benzyl-2-acetamido-3-O-carboxymethyl2-deoxy-G1-D-glucopyranoside dissolved in 40 ml of absolute dimethylformamide are added 3.87 g of L-alanine benzyl ester p-toluenesulfonate, 1.07 ml of N-methylmorpholine and 3.00 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline and the mixture is stirred for 24 hours at room temperature. The solvent is evaporated to give a monohydric residue which is worked up analogously to the procedure described in Example 31. After evaporation of the solvent and recrystallization from ethyl acetate, 4.5 g of benzyl-2-acetamido-3-O- [(L-carboxyethyl) -carbamoyl-methyl] -2-deoxy-α-D-glucopyranoside benzyl ester are obtained, m.p. 180-181 ° C, [α] δ ° = + 82 ° (97) ± 1 ° (methanol, c = 1), R f = 0.61 (system: acetonitrile / water 3: 1) and R f = 0. B1 (system: ethyl acetate / n-butanol / pyxidine / acetic acid / water = 42: 21: 21: 6: 10).

4.32 g of benzyl-2-acetamido-3-O- [(L-carboxyethyl) -carbamoyl-methyl] -2-deoxy-α-D-glucopyranoside benzyl ester, dissolved in 40 ml of ethylene glycol dimethyl ether / water 9: 1, are reacted for 2 hours hydrogen in the presence of 0.4 g of palladium coal. The catalyst is filtered off and the filtrate is evaporated to dryness. The residue is dried over phosphorus pentoxide to give 2.84 g, Rf = 0.47 (system: ethyl acetate / n-butanol / pyridine / acetic acid / water = 42: 21: 21: 6: 10), taken up in 40 ml of absolute dimethylformamide to which is added 2.18 g of D-iso-glutamine-Y-benzyl ester hydrochloride, 0.89 g of N-methylmorpholine and finally 2.16 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline. whereupon the mixture is stirred for 23 hours at room temperature. By working up in the same manner as described in Example 31, 3.95 g of benzyl-2-acetamido-3-O - <[L- (D1-carbamoyl-3-carboxypropyl) -carbamoylethyl] -carbamoylmethyl> -2-deoxy-α-D glucopyranoside benzyl ester m.p. 203-205 ° C, [a] §0 = + 66 ° (90) ± 1 ° (dime thyl formamide, c = 1,) Rf = 0.59 (system: acetonitrile / water = 3: 1),

Rf = 0.66 (system: methylene chloride / methanol / acetic acid).

Claims (9)

1. Analogous Process for Preparing Immunomodulatory 2- [2-Acylamino-2-Deoxy- (D) -glucosyl-3-O] -alkanecarboxylic Acid Peptide Derivatives of the General Formula wherein X represents a carbonyl group, R represents an alkyl group having from 2 to 17 carbon atoms or a phenyl group optionally substituted with alkyl of not more than 7 carbon atoms, R 2 means hydrogen, alkyl of not more than 7 carbon atoms or benzyl, R 2 hydrogen or alkyl having not more than 7 carbon atoms, R 4 and R q means hydrogen or alkanoyl having 2 to 18 carbon atoms, R 7 means hydrogen, alkyl having not more than 7 carbon atoms, hydroxymethyl or phenyl, R 8 means carboxyl, carbamoyl, N-C 1-7 alkylcarbamoyl, N-benzylcarbamoyl, N - (carbamoylmethyl) - carbamoyl or C1-7 alkoxycarbonyl, and Rg means carboxyl, carbamoyl, N-C1-7 alkylcarbamoyl or C1-7 alkoxycarbonyl, the dogalkyl group R containing more than one carbon atom when R 2 is methyl and R 2 and Rgh each represents a carboxyl group, or pharmaceutically acceptable salts of compounds with free carboxyl groups, characterized in that a) reacting a compound of the formula wherein X and R have the meanings set forth above and R 1a, R 40 and R g ° represent the above groups R 1, R 4 and R 9 or each a readily cleavable protecting group having a compound of the formula wherein Z represents a reactively esterified hydroxy group, R 2 has the meaning given above and R 70, R 8 ° and R g ° have the meanings given above for groups R 7, R 8 and R 9, however, carboxy and free hydroxy groups present in these groups are protected by easily split off protecting groups, and optionally split off any protecting groups present or b) condensing a compound of the formula wherein X, R and Rg have the meanings set forth above and Rx °, R40 and Rg ° have the meanings indicated for the groups R 1, R 4 and R g, or individually, means an easily cleavable protecting group or a reactive acid derivative thereof, with a compound having formula wherein R70, R8 ° and Rg ° have the meanings set forth above for the groups R7, Rg and Rg, however, these groups present carboxyl and, if desired, free hydroxy groups are protected with readily leaving protecting groups and leaving any protecting groups or groups present) condenses a compound of the formula wherein R4, Rx °, R2, R40, R6 ° ° R7 ° have the above meanings, having a compound of the formula wherein Rg ° and Rg ° have the above meanings, wherein, in the groups R7 °, Rg ° and Rg °, carboxyl and, if desired, free hydroxyl groups are protected with readily cleavable protecting groups, and cleave off any protecting groups present or d) in association with formula wherein R, R2, R70, 1 * 8 ° ° 9 ^ 9 ° are as defined above, and R5 is an alkylidene or cycloalkylidene group, under acidic conditions, the oxazoline and dioxolane ring cleaves, and optionally cleaves any protecting groups or e) in a compound of formula I wherein the substituents have the meanings set forth above, but functional groups present, if desired, are protected with readily cleavable protecting groups, acylating the amino group with an acylating agent such as introducer group R-X-, and optionally splitting off any present protecting groups or f) wherein R 1, R 40, and R 50 have the meanings set forth above for R 1, R 4, and R 9, respectively, or signify a readily leaving group, R 70, R 9, and R 9 have the meanings given above, respectively, R 7, R 9 and R 9, with carboxyl and hydroxy group may be protected with readily cleavable protecting groups, and X, R and R 2 have the meanings given above, however, with one compound of formula X, there is at least one readily cleavable protecting group, cleaving off the occurring protecting groups and, if desired, transferring one of the methods. a) to f) form a compound of formula I having at least one carboxyl group to a pharmaceutically acceptable salt thereof. Process according to claim 1, characterized in that starting compounds are used in which R 2 is hydrogen. Process according to claim 1 or 2, characterized in that starting compounds are used, wherein the hydrogen is hydrogen or an easily cleavable protecting group. Process according to one of Claims 1 to 3, characterized in that the starting compounds are used, wherein R 4 and R 4 represent hydrogen or an easily leaving group protecting group. Process according to one of claims 1-4, characterized in that starting compounds are used, wherein R is alkyl of 1-7 carbon atoms or phenyl. Process according to claim 1, characterized in that such starting compounds are used to prepare a compound of general formula I wherein. R is hydrogen or methyl of not more than 7 carbon atoms, hydrogen or methyl, R 4 and R 8 is hydrogen, R 7 is hydrogen, alkyl of not more than 7 carbon atoms or hydroxymethyl, R 9 is carbamoyl and R 9 is carboxy, however the alkyl group R contains more than one rona bona tom when R 2 is methyl, or a pharmaceutically acceptable salt thereof. Process according to claim 5, characterized in that such starting compounds are used to prepare a compound of formula I wherein R 1 represents hydrogen or a pharmaceutically acceptable salt thereof. Process according to claim 1 b) or c), characterized in that the starting compounds are selected to produce 2-acetamido-3-O - <[L- (D1-carbamoyl-3-carboxypropyl) -carbamoyl-ethyl ] -carbamoyl-methyl> -2-deoxy-D-glucose or a pharmaceutically useful salt thereof. Process according to claim 1 for the preparation of 2-acetamido-3-O - <[L- (D1-carbamoyl-3-carboxypropyl) -carbamoyl-ethyl] -carbamoyl-methyl> -2-deoxy-D-glucose or pharmaceutical useful salt thereof, characterized by reacting benzyl-2-acetamido-4,6-O-benzylidene-3- O-carboxymethyl-2-deoxy-α-D-glucopyranoside with L-alanyl-D-glutamic acid-1-amide-Y benzyl ester hydrochloride, decomposes the protecting groups into the compound and, if desired, transfers the compound formed into a pharmaceutically useful salt thereof.