CS205025B2 - Process for preparing derivatives of glukosamine - Google Patents

Abstract

Glucosamine derivatives of the formula <IMAGE> (X, R, R1, R2, R4, R6-R9 have the meaning stated in Claim 1) are prepared by condensation of corresponding 3-O-carboxymethylglucopyranosides with corresponding dipeptides. These glucosamine derivatives have the ability to increase the immunogenicity of an antigen on admixture therewith and, on systemic administration, to enhance the immunological reactivity of the treated organism. They can thus be used as adjuvants to increase the vaccine protection from bacterial, viral or parasitic pathogens.

Description

The present invention relates to a process for the preparation of novel glucosamine derivatives, in particular glucosamine-3-alkanoyl dipeptides, in particular compounds of formula I,

\ < ^{r X} C0-NH-CH-C0NH-CH (0) (I)

R 2 is hydrogen, C 1 -C 5 alkyl, hydroxymethyl or phenyl,

Re optionally an esterified or amidated carboxyl group a

An optionally esterified or amidated carboxyl group, provided that the alkyl moiety of R is more than 1 carbon atom when X is carbonyl and R2 is methyl, or when X is carbonyl, R2 is hydrogen and and R 9 represent a carboxyl group, as well as pharmaceutically acceptable acid addition salts thereof.

In the following, the term "lower" for radical or compound radicals means, unless otherwise indicated, preferably radicals containing up to 7, in particular up to 4, carbon atoms.

Alkyl is, for example, isopropyl, straight or branched, bonded in any position with butyl, pentyl, hexyl or heptyl, and in particular methyl, ethyl or n-propyl.

The optionally esterified or amidated carboxyl group is preferably the carboxyl group itself or the carboxyl group esterified with a lower alkanol such as methanol or ethanol in which

X a carbonyl group or a sulfonyl group,

R is C 1 -C 18 alkyl or phenyl,

R 1 is hydrogen or (C 1 -C 5) -alkyl or benzyl,

R 2 is hydrogen or C 1 -C 5 alkyl,

R4 and R6 are hydrogen or an acyl radical having 2 to carbon atoms,

Or a carbamoyl group which is unsubstituted on the nitrogen atom or is monosubstituted or disubstituted by an alkyl group, in particular a lower alkyl group, an aryl group, in particular a phenyl group, or an aralkyl group such as a benzyl group.

The carbamoyl group may also contain an alkylidene group such as a butylidene group or a pentylidene group. The carbamoyl group Ra may also contain a carbamoylmethyl group on the nitrogen atom.

In particular, the acyl group is an acyl group of an organic acid, in particular an organic carboxylic acid. Thus, the acyl group is in particular an alkanoyl group, in particular having 2 to 18 carbon atoms, such as acetyl or propionyl, or also an aroyl group such as 1-naphthoyl, 2-naphthoyl, and in particular benzoyl or halogen, lower alkyl, lower alkoxy, trifluoromethyl, a benzoyl or naphthoyl group substituted by a hydroxy or lower alkanoyloxy group, or also an acyl radical of an organic sulfonic acid, for example an alkanesulfonic acid, especially a lower alkanesulfonic acid such as methanesulfonic acid or ethanesulfonic acid, or an arylsulfonic acid, especially phenylsulfonic acid optionally substituted by lower alkyl benzenesulfonic acid or p-toluenesulfonic acid.

In the above compounds in which R 2 is an alkyl group, the acetic acid amide residue R 2 attached to the oxygen atom at the 3-position of the glucosamide moiety is optically active, i.e., occurs in the D-form. If R 7 is not hydrogen, the amino acetic acid R 7 is present in the L-form.

Depending on the nature of their substituents, the novel compounds according to the invention are neutral, acidic or basic compounds. When excess acid groups are present in these compounds, these compounds form salts with bases, such as ammonium or alkali metal or alkaline earth metal salts, for example sodium, potassium, calcium or magnesium. However, if excess basic groups are present, such compounds form an acid addition salt.

Acid addition salts are especially pharmaceutically usable, nontoxic acid addition salts such as inorganic acids such as hydrochloric, hydrobromic, nitric, sulfuric, or phosphoric acids, or organic acids such as organic carboxylic acids such as acetic acid , propionic, glycolic, amber, maleic, hydroxymalein, methylmalein, fumaric, cider, tartaric, lemon, benzoic, cinnamon, almond, salicylic, 4-aminosalicylic, 2-phenoxybenzoic, 2-acetoxybenzoic, em25bonic, nicotine or nicotine organic sulfonic acids such as methanesulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, ethane-1,2-disulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or naphthalene-2-sulfonic acid, as well as other acid addition salts, for example those which may be used as intermediates, for example ad for purification of the free compounds or for the production of other salts, as well as for characterization purposes, such as salts with picric acid, picrolonic acid, flavianic acid, phosphotungstic acid, fosfomolybdic acid, chloroplatic acid, Reinecke salt or perchloric acid salt.

The compounds according to the invention have valuable pharmacological properties, in particular they have a marked effect in potentiating immunity. This effect can be demonstrated by the following experiments: _r

1. In vivo potentiation of cellular immunity:

increased delayed type hypersensitivity to ovalbumin in guinea pigs

Guinea pigs (Pirbright) are immunized on day 0 with 10 mg of ovolbumin in complete Freund's adjuvant by injecting 0.1 ml of the mixture containing the adjuvant and antigen into both hind paws. Four weeks later, a skin reaction is induced by intracutaneous injection of 100 μ% ovalbumin in 0.1 ml of buffered saline and a quantitative evaluation is made after 24 hours based on the reaction volume calculated using the area of erythema and the increase in skin thickness. The antigen-specific increase in reaction volume observed after 24 hours (delayed reaction) is a measure of cellular immunity. Ovalbumin is too weak an immunogen to induce a delayed reaction either alone or in a water-in-oil emulsion with incomplete Freund's adjuvant (10 parts ovalbumin solution) in 0.9% sodium chloride solution mixed with 8.5 parts Bayol F (mineral oil) and 1.5 parts of Arlacel A (mannitol monooleate), and for effective immunization, must be added in complete adjuvant to which mycobacteria (5 mg killed and lyophilized M. butyricum per 10 ml Bayol F /) have been added. To demonstrate efficacy in potentiating the immunity of test substances, these substances may be admixed with mycobacteria at doses ranging from 10 to 100 μξ to the antigen-in-oil mixture.

The glucosamine peptides of this invention have the ability to perfectly mimic the effect of mycobacteria in the described experimental setup and quantitatively exceed it.

Statistically significant potentiation of the delayed response to ovalbumin can also be achieved by not adding the compounds of the described type to the antigen-in-oil mixture, but administering subcutaneously at doses of 10 to 100 μξ per animal within a few days after immunization (e.g. , 2, 5, 6 and 7) in sodium chloride solution.

This demonstrates that compounds of the type are capable of substantially enhancing cellular immunity, both in admixture with the antigen alone (adjuvant effect in the strict sense) and in a time and locally separate delivery from the antigen injection (systemic potentiation of immunity).

2. In vivo potentiation of humoral immunity: increased production of bovine serum albumin antibodies in mice

Mice (NMRI strain) are immunized by intraperitoneal (ip) injection of 10 μ% clot-free bovine serum albumin on day 0. For the 9, 15 and 29 days was sampled and found serum antibodies to albumin serum by _e vězího passive hemagglutination. At the dose used, the soluble bovine serum albumin is subimmunogenic to the animals to which it is administered, i.e., it is unable to elicit any, or induce only very low production of antibodies. Additional administration of certain immunopotentiating agents to mice before or after antigen administration results in an increase in serum antibody titer.

The effect of this administration is expressed by the value of the degree achieved, ie the sum of the log2 titer differences on the three days of blood collection.

The compounds of the invention are capable of increasing statistically significant production of antibodies against bovine serum albumin when administered intraperitoneally or subcutaneously at a dose of 100-300 mg / kg animal weight for five consecutive days (i.e., days 0-4) after immunization with bovine serum albumin.

The immunostimulatory effect of these compounds is antigen-dependent in contrast to other bacterial immunoleptics (e.g. LPS from Escherichia coli). Injection of the new compounds causes an increase in antibody titer against bovine serum albumin only in mice that have been immunized with bovine serum albumin, but not in mice that have not been immunized. It is noteworthy that the subcutaneous dose of the compounds is as effective as the intraperitoneal dose, i.e. the observed immunopotentiating effect is systemic and is not dependent on whether the stimulant is administered in the same way as the antigen or must be administered. together with the antigen, as is the case with classical adjuvants.

The experiments described above demonstrate that compounds of the type described are capable of specifically enhancing humoral immunity, that they improve the immunological response to irritation, and that their immunopotentiating effects are based on systemic activation of the immune system.

3. In vitro potentiation of humoral immunity:

effect substituting effect of T-cells in response to antibodies in mouse spleen cells on sheep erythrocytes (SE)

In many cases, thymus lymphocytes (called T-cells) are required to induce antibody responses. These cells cooperate with emerging antibody-forming lymphocytes (B-cells) to help them respond to stimulation by so-called T-dependent antigens while simultaneously proliferating, differentiating, and synthesizing antibodies. Spleen cell suspensions of congenitally athymic nu / nu mice do not contain any functional T cells and, for example, cannot produce any antibodies against sheep erythrocytes in vitro in the presence of sheep erythrocytes. Surprisingly, the compounds of the present invention are able to functionally replace T cells in such cultures and thus allow antibody response against sheep erythrocytes by antibody production. The addition of these substances to nu / nu cultures of spleen cells in the presence of sheep erythrocytes leads to a significant increase in the number of antibody-forming cells within 4 days. The results show that the compounds are able to increase the production of humoral antibodies in vitro and to compensate for T cell deficiency in the system.

4. Selective B-cell mitogenicity: effect on · stimulation of proliferation in B-lymphocyte cultures

Suspension of highly enriched B cells (lymph node cells of congenitally athymic nu / nu mice), as well as largely pure immature or mature T cells (thymus cells or thymus cells from Balb / c mice, cortisone-resistant cells, i.e. cells). persistent 48 hours after cortisone injection) are incubated for 3 days in the presence of test compounds. Incorporation of H ³ -thymine into lymphocytes during the last 18 hours of culture is considered a measure of proliferative activity.

The compounds of the invention are mitogenic to B cells (ie, precursors to antibody-forming cells), but not to T cells.

Thus, they are capable of promoting the proliferation of lymphocytes involved in humoral immunological responses.

5. Tolerance

Although the compounds of the described type show potentiating effect in guinea pigs, for example after a single dose of 0.05 mg per kg by subcutaneous administration, and in mice given five doses of 10 mg per kg subcutaneously, they were not observed in mice even at five doses after administration. 300 mg / kg intraperitoneally no toxic effects. They therefore have an excellent therapeutic range.

The compounds of the present invention are capable of being mixed with antigen 205025 on the one hand

R ~ C ~ H \ -.

10-NH-CH-CONH-CH (CCyf enhances its immunogenicity, on the other hand, increases the immunological reactivity of the treated organism when administered systemically, while the compounds are capable of promoting both cellular and humoral immunity and activate lymphocytes that form antibodies .

Accordingly, the novel compounds may be used as adjuvants in admixture with vaccines to improve the vaccination result and thereby protect against infection mediated by humoral and / or cellular immunity against bacteria, viruses or parasites.

Furthermore, the compounds are useful in admixture with a variety of antigens as adjuvants in the experimental and industrial production of antisera for therapy and diagnosis and for inducing immunologically activated lymphocyte populations in cellular transfer.

In addition, the novel compounds can be used without the simultaneous supply of antigen to support already weak immunological reactions in humans and animals. Accordingly, these compounds are particularly useful for stimulating the body's own defense, for example in chronic and acute infections or in selective (antigen-specific) immunological defects, as well as inherited, but also after generally acquired [i.e. not antigen In general, immunological defective conditions as may be encountered in old age, during more severe primary diseases, and in particular after ionizing radiation therapy or hormones that act to suppress immunity. These substances can therefore be s. advantageously also given in combination with antiinfective. antibiotics, chemotherapeutics or others. methods of treatment to counteract immunological damage.

Finally, the compounds are also useful in the general prophylaxis of infectious diseases in humans and animals.

In particular, the invention relates to a process for the preparation of compounds of formula (I), in which:

X is a carbonyl group,

R is lower alkyl or phenyl and

R1, R2, R4, R6, R7, Rb and Ro are as defined above, and salts thereof.

Of particular interest are also compounds in which R2 is hydrogen and the other radicals are as defined above, and salts thereof.

Particular emphasis should be placed on the compounds of formula (ii).

(II) in which

R is lower alkyl or. phenyl group,

R 1 is hydrogen or lower alkyl,

Rz ·. hydrogen or methyl,

R 7 is hydrogen, C 1 -C 5 lower alkyl or hydroxymethyl,

Re is a carbamoyl group and a

Ra is a carboxyl group, provided that the lower alkyl moiety R is. contains. more than 1 carbon atom when R 2 is methyl; salts. ;

In particular, the compounds of the general formula (II) in which they are represented are to be mentioned

R is a lower alkyl or phenyl group,

R 1 is hydrogen,

R 2 is hydrogen or methyl;

R 7 is hydrogen, methyl or. hydroxymethyl group,

Ra is a carbamoyl group and a

R 9 is a carboxyl group, provided that the lower alkyl moiety of R has more than 1 carbon atom when R 2. represents a methyl group, and salts thereof.

The novel compounds of formula (I) and their pharmaceutically acceptable acid addition salts. according to the invention are produced by the compound of formula III,

CHgORt

^-R 1 (II) nh-xR

COOH in which

X, R, and R2 are as defined above and

R 10, R 9 and R 6 have the meanings of the radicals R 1, R d and R 6, respectively, or are readily cleavable capping groups, or a derivative of this compound is condensed with a compound of formula IV,

R7 ° · Rs °

II

IHN-CH-CONH-CH (CH2) 2-.Rg ^of (IV) in which

R ⁷⁰ , R 8 and R 9 are as defined in R 7, R 8 and R 9, except in these radicals. . the carboxyl groups present and optionally the free hydroxyl groups are protected by readily cleavable capping groups, whereupon the optionally present capping groups are cleaved, whereupon any salts obtained are converted into free compounds or the salts obtained are converted into their physiologically used acid addition salts.

The coupling is carried out, for example, by reacting the compound of formula III in the form of an activated carboxylic acid with an amino derivative of formula IV or reacting the acid of formula III with a compound of formula IV whose amino group is present in the activated form. The activated carboxyl group may be, for example, an acid anhydride, preferably a mixed acid anhydride, such as an acid azide, an acid amide, such as an imidazolide, -isoxazolid or an activated ester. Activated esters include, but are not limited to: (cyanomethyl ester, carboxymethyl ester, p-nitrophenylthioester, -p-nitrophenyl ester, 2,4,5-richlorphenyl ester, pentachlorophenyl ester, N-hydroxysuccinimide ester, N-hydroxyphthalimide ester, 8-hydroxyquinolmester, 2-hydroxy- 1,2-dihydro-1-ethoxycarbonylquinolinester, N-hydroxypiperidinester or enol esters obtained with N-ethyl-5-phenylisoxazole-3'-sulfonate.

Activated esters may also be obtained optionally by treatment with carbodiimide with the addition of N-hydroxysuccinimide or unsubstituted or, for example, halogen-substituted methyl or methoxy-substituted 1-hydroxybenzotriazole-3-hydroxy-4-oxo-3,4-dihydrobenzo [d]. -1,2,3-triazine.

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

Among the methods of reaction with '-activated esters, mention may be made in particular of N-ethyl-5-phenyl-soxazolium-3'-sulfonate (Woodward's reagent K) or 2-ethoxy-1,2-dihydro-1-ethoxy (carbonylquinoline or carbodiimide.

Easily cleavable capping groups are those known from peptide chemistry or sugar chemistry. For carboxyl groups, in particular, tert-butyl, benzyl or benzhydryl and, for hydroxyl groups, in particular acyl groups, for example lower alkanoyl groups such as acetyl, aroyl groups such as benzyl, and in particular residues derived from - carbides such as benzyloxycarbonyl or alkyl, especially tert-butyl, optionally nitro, lower alkoxy or halogen substituted benzyl or tetrahydropyranyl, or optionally substituted alkylidene radicals, which are bonded to the oxygen atoms at the 4 and 6 positions. Such alkylidene radicals are in particular a lower alkylidene radical, in particular an ethylidene radical, an isopropylidene radical or a propylidene radical or alternatively an optionally substituted, preferably in the p-position, substituted benzylidene radical.

These capping groups can be removed in a manner known per se. Thus, they can be removed by hydrogenolysis, for example with hydrogen in the presence of:. a noble metal catalyst such as a palladium or platinum catalyst, or acidic hydrolysis.

The starting materials used are known or can be prepared in a manner known per se. Thus, compounds of formula III can be obtained, for example, by reacting the corresponding 3-position of unsubstituted sugar with halo-R 2 -acetic acid, wherein R 2 is as defined above, and its esters in the presence of a strong base. Halogen is preferably - bromine - or especially chlorine.

The compounds obtained can be converted into their salts in a known manner, for example by reacting the acid compounds obtained with alkali metal hydroxides or alkaline earth metal hydroxides or by reacting the obtained basic compounds with acids.

The processes described above are carried out by methods known per se, in the absence or preferably in the presence of a diluent or solvent, optionally with cooling or heating, under elevated pressure and / or under an inert gas atmosphere such as nitrogen.

In this connection, all substituents present in the molecule, in particular in the presence of readily hydrolyzable O-acyl radicals, can be used in particular: - gentle reaction conditions, - such as short reaction times, use of slightly acidic or basic reagents at lower concentrations, stoichiometric ratios, select suitable catalysts, solvents, temperatures or pressure conditions.

The invention also relates to those embodiments of the process, wherein the starting material is a compound obtained as an intermediate at any of the steps of the process, and the missing reaction steps are carried out, or the starting material is formed under the reaction conditions, or used in the form of a reactive derivative or a salt which, according to the invention, lead to the compounds described above as being particularly valuable.

The invention also relates to pharmaceutical compositions comprising compounds of the formula

0'5‘0'2 r

I. The pharmaceutical preparations according to the invention are enteral preparations; as orally or rectally, as well as for parenteral administration to a warm-blooded animal, which comprise a pharmacologically active agent alone or together with a pharmaceutically acceptable carrier. The dose of the active ingredient depends on the species of warm-blooded individual, age and individual condition; as well as the route of administration:

The novel pharmaceutical preparations contain from about 10 to about 95%, preferably from about 20 to about 90%, of the active ingredient. The pharmaceutical compositions of this invention may be in the form of dosage units, such as dragees, tablets, capsules, suppositories, or ampoules.

The pharmaceutical compositions of this invention also has prepared by processes which are known per se ^1, for example using conventional · Mixing, granulating; by draining, dissolving, or lyophilizing processes. In addition to the methods of using the above-mentioned page 13, pharmaceutical compositions for oral administration may be prepared; when the active ingredient is mixed with a solid carrier, the mixture obtained is granulated, after which the mixture or granulate, if desired or necessary, is further processed after addition of suitable excipients to tablets or dragee bases;

Suitable carriers are, in particular, fillers, such as sugars, for example lactose, sucrose, mannitol or D-glucitol, cellulose and / or calcium carbonates, such as tricalcium phosphate or calcium hydrogen phosphate ¹ , as well as binders such as starch wax using e.g. maize, wheat, rice or potato starch, gelatin, tragacanth, · methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose and β-pilyl vinylpyrrolidone; and / or - if desired ¹ - disintegrators, such as the abovementioned starches, furthermore karboxyrtiethyiškrob, crosslinked polyviiiylpyrrolidon, agar, alginic acid or salts thereof such as sodium salt and the auxiliary 'means jsdu primarily regulating agents fluidity; Furthermore, the pharmaceutical preparations contain ^one lubricant 'as' kyseliúa silica; talc, stearic acid or salts thereof such as magnesium or calcium salt and / or polyethylene glycol. The dragee bases are coated with a suitable coating or gastric juice-resistant coating, using, inter alia, concentrated sugar solutions which optionally contain gum arabic, talc, polyvinylpyrrolidine; polythylene glycol and / or titanium dioxide, as well as solutions of lacquers in suitable organic solvents or mixtures of orthopharmic solvents or in the preparation of gastric juice-resistant coatings, solutions of suitable cellulose preparations such as cellulose acetyl phthalate or liydroxypropylmethyl phthalate methyl phthalate are used. Dyestuffs or pigments may be added to the tablets and dragee coatings, for example for identification purposes or to indicate different doses of the active ingredient.

The following examples are intended to further describe the invention without, however, limiting it thereto. Temperature values are given in degrees Celsius;

Example 1

A solution of 2.6 g of benzyl 3-O- {D1- [L1- [D1-carbamoyl-3-carboxypropyl) carbamoylethyl] carbamoylethyl} -2-deoxy-2-proplonylamino-α-D-glucopyranoside benzyl ester in 67 ml 60% acetic acid is hydrogenated at normal pressure and at room temperature after addition of 0.6% palladium on carbon as catalyst for 0.6 hours until hydrogen consumption ceases (about 20 miri.). The catalyst is filtered off, washed with a small amount of 60% · acid. The residue is recrystallized from a mixture of ethanol and ether to give benzyl-3O-N-1- [L1- (D1-carbamoyl-3-carboxypropyl) carbamoylethyl] carbamoylethyl] carbamoyl-ethyl. Z-deoxy-Z-propionylamino-α + D-glucopyranoside, mp 155-160 ° C (decomposed) and * of optical rotation [α] D = + 10 5 + 1 ° (dimethylformamide, c = 0, 58).

Benzyl 3-O- [D-1- [L-1- (D-1-carbamoyl-3-carboxypropyl)] carbamoyl-letlyl] benzyl-carbamoylethyl-Z-deoxy + Z-propiohylamino-α-D-glucopyranoside used as the starting compound; can- prepare! thus:

4.4 g of benzyl-4-amino-.alpha.-benzyl-S-O + D-4-carboxyethyl) -2-deoxy-.alpha.-D-glucopyranoside are dissolved in 200 ml of methanol in the presence of 3 ml of triethylamine. with stirring at room temperature, 0.95 ml of propionic acid chloride is added dropwise to the solution. The reaction mixture is stirred for 2 hours, then concentrated to dryness and the residue is dissolved in 100 ml of water. Add 2 N hydrochloric acid solution, cooled in ice; The solution is adjusted to pH 4, the precipitated product is filtered off with suction, washed with water and dried in vacuo over sodium hydroxide. The thus prepared benzyb4-6-Q-benzylidene-3-Q- (D1-carboxyethyl) -2-deoxy-2-proplonylamino-1H-glucopyranoside can be recrystallized from methanol, mp 257 ° a * [1a] _D ²⁰ = = -1-132 ^ + 1 ^ (dimethyiformamide; c = 1; 086)

To a solution of 3.1 g of the compound thus obtained in 150 ml of atetonitrile and 0.9 ml of triethylamine, 1.5 g of N-ethyl-5-phenyl-isoxazolium-3'-sulfonate (Woodward reagent К) was added and The mixture is stirred at room temperature until complete dissolution of all components, which takes about 60 minutes. 2.2 g of L-alanyl-D-glutamic acid β-benzyl ester 1-amide hydrochloride and 0.9 ml of trlethylamine were added and the reaction mixture was stirred at room temperature for 18 hours. The solvent is distilled off under vacuum of a water pump, water is added to the residue, the insoluble matter is filtered off with suction, thoroughly

015: 0> 2 ^! 5>

washed with water and dried; Thus obtained · · BEM zyleste r · BE džyI ^ ^ - ^, ^(j ^ (^ -t ^ e ^ r ^ 2 ^ z ^ Lider-3-O- {I ^ --.- [L 14D-β-Bamyl-3-carboxypropylcarbamoyl-ethyl] carbamoylethyl-2-deoxypropylpropionylamino-di-glucopyranoside having an Rf value of 0.45 on thin layers of silica gel using a 10/1 methylene chloride / methanol mixture. 60% acetic acid and the solution is heated at 95-100 ° C for 1.5 hours After cooling, the solvent is distilled off, the residue is dissolved twice in a small amount of water each time. was distilled off · · · · each to-dry) and then a · thus - obtained 'benzyl benzyl-S ^ O-Jd ^1! - · [· i Ί- (D where ^ З Ikaгbamoy ⁱ karbotfУpropyУ-ethyl jlaτbamoylI Carbamoylethyl-2-videoxy-12-propionyl amino-2D-glucopyranotide after crystallization from methanol, mp 189 ° and [α] D ²⁰ - + 10 (0 + 1 ° [d R methylformamide, c = 0.268).

Example 2

Solution · 4 · g · Benzyl ester · Benzyl-2-acetonitrile to jiji--1-ID (carbamoyl-3-carboxypropyl-1-yl) -carbamethylethyl} -carbamoylmethyl} -deoxyde-ilucopyranosicide in 80 ml of methanol with 80 ml of methanol The addition of O-4 'g' 5% 'palladium on carbon as a catalyst hydrogenates at normal pressure and temperature ^; room until hydrogen is consumed. The catalyst is then filtered off, washed with a small amount of methanol and the filtrate is concentrated to dryness in a vacuum pump. The residue is dissolved in 50 ml of distilled water, 1 g of 5% palladium on carbon catalyst is added and the solution is hydrogenated at normal pressure and at normal temperature until the consumption of hydrogen is complete. The catalyst is filtered off with suction, washed with a small amount of water and the filtrate is concentrated to dryness. The thus-obtained 2-acetamido [10-a] {[1-li [D: 14 (arbamoyl-3-carboxypropyl)] carbonyl] -yl-methyl} -cyclohexyl} -cyclohexyl-2-acetamido-3-carboxypropylene The I / O is dried under high vacuum over phosphorus pentoxide. · The substance has a specific rotation Mn ²⁰ = з-00 ^о + 1о (water, c = 0.930).

The starting compound is prepared as follows: To a solution of 9.5 g of renzyl 12 -acetamido-4,6-α 1 -ellyl-1-yl-3-O-carboxymethyl-12-deoxy-α-D-glucocyraacetide in 400 ml of acetic acid. ii ^ i ^^^ of clay and 3 ml of triethylamine · · are added 5.3 g hyR5 Nieto-fenylieoxazollumi3 ^of ItulfOnátu [..WcCwardovo reagent · · K), and the reaction mixture · · stirred 'za-' at room temperature until · 7.15 g of hydrochloride, 1-amide, 1-amide, 1-allyl R-glutamic acid, 3 ml of triethylamiil and 200 ml of acetomtril are added to the solution, and the reaction mixture The mixture was stirred for an additional 18 hours at room temperature. The portion of rehzyl-1,2-acetamido-4,6-O-benzylidene-3-O-1- [L, -1- [1-D-1-carbamoyl-3-larroxypropyl] carbamoylethyl] carramoylmethyl] -2-oxyoxy-D-g-lucopytanoside, which crystallizes; aspirate, wash with half-saturated sodium bicarbonate solution in water and dry; the substance has a specific rotation [α] _D 2 ° = + 8Г + Г (dimethyfiormamide, c = 0.816).

A solution of 8 g of this compound in 409 ml of 60% acid solution. The acetic acid was heated at 89 ° C for an hour. After cooling to ·· · '· sodium · concentrated to dryness, the residue was twice · ¹ · · added with 50 ml of water each and the reaction mixture was concentrated, · · to the' dry ·· · ,, residue which follows · · yield in the crystalline state, it is stirred into a small amount of water, the crystals are suctioned off and dried. · benzyl ester of benzyii2iaceti amid0-3iOi {[· L · lkarbamoyRЗ ^{14D-i-propyl-lj} kar.boxyi carbamoyl-y ^ · ^ methylethyl karramoylmethyllI I2> de oxy-Dggukopyyranotl · · · du ol mp 200 DEG · [AJD ' ^{2 (} 0 = · + 77 ° + 1'-dimethylformamide, c = -0,599): ”

Snakes 3 ·

A solution of 0.9 g of benzyl ester rehzyl-2-acetyl-rnido-3-Ο- [+ 1- · 1D + carbamoyl-3-larboxypropyl) carbamoylthyl] carbamoylmethyl-2-deoxy-6-O-ether -oyl-α-D-glucopyranoside in 40 ml of methanol, add 0.2 g of palladium? on; The coal as a catalyst is hydrogenated at normal pressure and at normal temperature until consumption of 22.4% by weight of hydrogen. After filtering off the catalyst, which is washed with methanol; The liquid fraction is concentrated to dryness to give benzylhazzamido-3-ol [L-11 D + carbamoyl-3-carboxypropyl] carbamoylthyl] -carbamethylmethyl-2-deo-1-iodo-stearyl-IaD- 3-Όpyranosl · cl-o [:]] d 2 ° ~ = + 33 ° + 1 ° (chloroform, c = 1.046).

The starting compound is prepared as follows: 2.8 g <behzyletter ^, benton-2-acetamido * 3iO1- [^^^ 1] 1D-1 ^ '- 1 ^^^ 1 ^ 1 ^^ allyl propyl (i) carbamethylethyl] carbamoylmethyl-2-deo-xyza-D-ghopylgahOSide in 30 ml of pyridine was stirred with stirring. aiza exclusion of moisture! a solution of 1.4 is added dropwise over an hour; grams of chloride. Stearic acid in 7.5 ml of methylene chloride and the reaction mixture is stirred for 40 hours at room temperature. The reaction mixture was then poured into ice water and extracted with methylene chloride. The organic portion is washed with 2 N aqueous hydrochloric acid solution, cooled with water, then dried with water; with anhydrous magnesium sulfate and the filtrate is concentrated to dryness. The residue is purified by chromatography on a column of silica gel, eluting with a silica gel; of acetic acid.

There was thus obtained benzyl ester of benzyl-2-acetamycin-2-oxo-2-di-1-larbamoyl-3-carboxypropyl) carbamoylmethyl-carbamoylmethyl-1,2-deoxy-4-thienoid-a-D-glucopyranoside o. rotation · [- + ²⁰ = ^; +30 ^ο + 1 ° (chloroform, c = 1.203).

Example 4

Similarly, according to the procedure of Example 2, rehzyl-1,2-acetamido-4,6,6-O-benzylidene-10- (D-1-methyl-4-propyl) - 12-oxyoxy-1-D-glucopyranoside was condensed with hydrochloride. 1-amide z-ester of L.-glym-D-glutamic acid and the protecting groups are cleaved off; Gets.

2-acetamido-3-O- (D 1 - [(D 1 -carbamoyl-3-carboxypropyl) carbamoylmethyl] carbamoylpropyl] -2-deoxy-D-glucose. [α] d ²⁰ = = -1-460 + 1 ° (H 2 O, c = 0.630).

The starting compound is prepared as follows:

To a solution of 60 g of benzyl-2-acetamido-4,6-O-benzylidene-2-deoxy-α-D-glucopyranoside in

600 ml of dimethylformamide were added with 10 g of sodium hydride and the reaction mixture was stirred under nitrogen at 45 ° C for 1.5 hours. After cooling to 0 ° C, 75 ml of DL-α-bromobutyrate are added, and the reaction mixture is stirred at room temperature for 1 hour, then at 50 ° C for one hour, neutralized. followed by acetic acid, whereupon the solvent was distilled off under water pump vacuum. The residue was partitioned between methylene chloride and water, the organic phase was washed once more with water, and after drying over magnesium sulfate, the solvent was distilled off. The residue, dried under high vacuum, is purified by silica gel column chromatography. Elution with mixtures of methylene chloride and ethyl acetate (85/15) affords ethylbenzyl-2-acetamido-4,6-O-benzylidene-3-O- [D1-carboxypropyl] -2-deoxy- and D-glucopyranoside ethyl ester of specific rotation [α] D = = 113 ° + 1 ° (chloroform, c = 0.5). After crystallization from methylene chloride / ether, mp 154 ° C, Rf 0.21; furthermore gave ethyl benzyl 2-acetamido-4,6-O-benzylidene-3-O- (Ll-karboxypropyy) -2-deoxy-D-glucopyranoside of the value of specific rotation [ «) d ² 0 - + 42 °° +1 (chloroform, c = 0.511), mp 240 (crystallized from ethyl acetate): Rf value = 0.04.

To a solution of 38.1 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O- [D-1-carboxypropyl-2-deoxy-α-D-glucopyranoside ethyl ester in

300 ml of methanol are added with 100 ml of 1 N sodium hydroxide solution and the reaction mixture is kept at 60 ° C for 1 hour. After cooling, the solution is concentrated to a volume of approximately 150 ml. The concentrate was diluted by the addition of 400 ml of ice-water and 100 ml of 1N ice-cooled hydrochloric acid was added. The product which crystallizes is filtered off with suction, washed with water and dried. There was thus obtained benzyl-2-acetamido-4,6-O-benzodiazole-3-O-D-carboxymethyl-2-deoxy-α-D-glucopyranoside of m.p. 210 DEG-213 DEG C. and (a) _n ²⁰ = + 11 (0 ± 1 ° (dimethylformamide, c = 0.55-4 ')).

Similarly, benzyl-2-acetamido-4,6-O-benzylidene-3-O- (11-carboxy-propyl) -2-dnoxy-α-D-glucopyranoside is obtained having a melting point of 285 ° C and [α] ο ²⁰ = + 71 DEG (dimethylformamide, c = 0.589).

Example 5

Similar to Example 1, ethyl-2-benzamido-2-deoxy-3-O-carboxymethyl- + D-glucopyranoside and α, γ-bis-methyl-4-aminohexanoic acid L-alanyl-D-glutaimic acid hydrochloride were obtained from ethyl 1-2-eenzamide-2-deoxy-3-O- [11- [D 1,3-bis-N-methylcarbamoylpropyl) carbamoylethyl] carbamoylmethyl-β-D-glucopyranoside, mp 233-240 ° C, [a) D ²⁰ -20 ° (methanol, c = 0.937). TLC (silica gel, 7: 3) _Rf 0.38.

Example 6

Analogously to Example 1, 2-benzamido-2-dnoxy-0-O- [L-1- (D1, -is-N-methylcareamoylpropyl)] was obtained from 2-benzamido-2-deoxy-3-O-carboxymethyl-D-glucopyranose. kareamoylnthyl-D-glucopyranose by? · 125 to 132 DEG C. [л] - ^Я). = + 24 DEG (H2O, c = 0.93), _Rf value = 0.26 when chromatographed on a thin pad of silica ^ agel in chloroform-ethanol (7: 3).

Example 1 a d 7

Similarly to Example 1, 2-benzamido-2-deoxy-3-O- [L1-] is obtained from 2-eenzamido-2-deoxy-3-O-carboxymethyl-D-glucopyranose and L-alanyl-D-glutamic acid dl-methyl ester hydrochloride. (Dl, 3-bis-methoxykareonylpropyl) carbonyl bamoylmethyl] carbamoylmethyl-d-glucopyranose, as a hydrate, mp 80-90 °, [ «] d ² 0 = + 25 ° (methanol, c = 1.017). TLC in silica gel (9: 1) showed Rf = 0.23.

Example8

Analogously to Example 1, ethyl 2-benzamido-2-deoxy-3-O- [L1- (D-) -O-carboxymethyl-N-D-glucopynganoside is obtained from ethyl-2-enzamido-2-dnoxy-0-O-carboxymethyl-N-D-glucopyranoside. 13-bis-methoxycarbonylpropyl) carbamoylethyl] carbamoylmethyl- + D-glucopyranoside mp 127-135 ° C; [.Alpha.] _D @ 20 = -17 DEG (CH3 OH, c = 1.0-24). The substance had an Rf of 0.26 when thin-layer chromatography on silica gel with ethyl acetate-acetone (2: 1).

Example 9

Analogously to Example 1, 2-benzamido-2-deoxyl-2- [L-1- (D-1-N-carbamoylmethylcareamoyl-6-carboxyoxypropyl) carbamoylmethyl] carbamoylmethyl-D-glucopyranose was obtained. The N-tert-butoxycarbonyl-L-alanyl-D-glutamic acid 7-benzyl ester and α-glycinamide of the peptide moiety are used as starting compounds. M.p. 163-170 ° C.

Example 10

Similarly to Example 1, 2-benzamido-2-deoxy-3-O- [L1- (D1-careamoyl-0-carboxypropyl) 2-benzamido-2-deoxy-3-O- [L1- (D1-careamoyl-0-carboxypropyl)] was obtained from α-amide α-amide x-tert-butyl ester hydrochloride. carbamoylbutyl] carbamoylmethyl-D-glucopyranose. Mp 114-115 ° C;

[Α] -2 ° = + 34 + 4 ° (c = 0.89, H 2 O).

Example 11

Analogously to Example 1, 2-benzamido-2-deoxy-3-O- [11- (D1-carbainoyl-3-carboxypropyl) carbamoyl-2-methylpropyl] -1-carbamoyl ^} methyl] -D [glucopyranose was obtained.

Rf = 0.3-5; (Chloroform / ethanol 1: 1).

Example 12

To a solution of 10.7 g of benzyl 2-acetanildo-1-O-benzylidene-4-Of (D 1 -carbamoyl) -3-carboxypropyl) carbamoylmethylcarbamoylmethyl iodoxy-α-D-glucopyranoside benzyl ester in 200 ml glacial acetic acid and 100 ml 2 g of 5% palladium on carbon are added to the water, and the reaction mixture is hydrogenated for 57 hours at atmospheric pressure and room temperature. The catalyst was filtered off, washed with water and the filtrate was concentrated. The residue was dissolved in water, the solution was filtered through 100 ml of Amberlit IR 120 ion exchange resin in the H + -cycle, and the filtrate was lyophilized. 2-Acetamido-3-O - [(D1-carbamoyl-3-carboxypropyl) carbamoylmethylcarbamoylmethyl] -2-deoxy-D-glucose was crystallized from a mixture of methanol and ethyl acetate. Drying under high vacuum yields a product containing 1/4 mol of ethyl acetate having a specific rotation of [.alpha.] D @ ²⁰ = + 27 DEG. + -. 1 DEG (water, c = 0.944).

The starting compound used in the process is prepared as follows:

g of N-tert-butyloxycarbonylglyeyl-D-isoglutamic acid benzyl ester is dissolved at room temperature and free of moisture in a mixture of 18 ml of 1,2-chloroethane and 8.4 ml of trifluoroacetic acid, and the reaction mixture is allowed to stand for 16 hours. The reaction mixture is diluted with 200 ml of tetrahydrofuran, neutralized with external triethylamine under external cooling, and a solution of 8.3 g of benzyl-2-acetamido-4,6 [O-benzylides-P-O-carboxymethyl-4-deoxy] is added. -az-D-gliicopyranoside in 100 ml of tetrahydrofuran and 2.52 ml of triethylamine. After a further addition of 5.05 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline, the reaction mixture was stirred at room temperature for 24 hours. The product which crystallizes is filtered off with suction, washed with tetrahydrofuran and ether and then dried. The benzyl 2-acetamido-4,6-O-benzylidene-3-O - [(D1-carbamoyl-3-carboxypropyl) carbamoylmethylcarbamoylmethyl] -2-deoxy- _[ ac] _D -glucopyranoside benzyl ester thus obtained has a specific rotation of [α] _D 2 [Α] D = + 66 ° C + 1 ° [N, N-dimethylformamide, - c = 1.380].

Example 13

A solution of 4.5 g of methyl 2-acetamide benzyl ester [3-O- [[1- (D1 [carbamoyl-3-carboxypropyl) carbamoylethyl] carbamoylmethyl] -4-deoxy-α-D-glucopyranoside in 125 ml of 50% aqueous methanol is added. addition of 1.0 g of 5% palladium on carbon is hydrogenated under normal pressure at room temperature until 178 ml of hydrogen are consumed. The catalyst is then filtered off and the filtrate is concentrated under a water pump vacuum. The residue was dissolved in ~ 50 ml of distilled water and the solution was lyophilized. Yield - was thus obtained methyl 2-acetamido-3-O - {[L-l (DL-kaτbamoylc -karboxypr-PROPYL) carbamoylethyl] carbamoylmethyl | -2-deoxy-CRD-glllkopyranosid axis ^P ec ^{i a} l FIC rotation [ ] ⁰²⁰ = + 4 ° + ⁹ ° l ^[vO'da, c = 0.939).

The starting compound is prepared as follows:

8.1 g of N-tert-butyloxycarbonyl [L-alanyl [D-isoglutamic acid benzyl ester] are dissolved in a mixture of 8.1 ml of tert-butylacetate and 8.1 ml of trifluoroacetic acid at room temperature and free of moisture. 16 hours. The reaction mixture was diluted with 203 ML of tetrahydrofuran, neutralized with triethylamine, with external cooling and the reaction -Mixtures -se solution of 7.62 g of methyl 2-acetamido-4,6-O-benzyli'de! Nc3cO- karboxχmethyll2 ^L-ιdeoxy α-D-glUc of copyranoside and 2.77 ml of triethylamine in 100 ml of tetrahydrofuran. After addition of 5.0 grams of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline, the reaction mixture is heated to 40 ° C, stirred at this temperature for 30 hours, and then allowed to stand for an additional 24 hours at rooms. Excluded product, ie methyl 2- (a) etamido-4,6-O-benzylidene-3-O - {[L1- [D4-carbamoyl-3-carboxypropyl) carbamoylethyl] carbamoylmethyl} -2-benzyl ester The deoxy- [alpha] -D-glucopyranoside is filtered off with suction, washed with tetrahydrofuran and ether and then dried. The substance has [α] ²⁰ D = + 58 ° + 1 ° (N, N-dimethylformamide, c = 1.125).

A solution of 10.5 g of methyl 4-acetamide O [4,6-O-benzylidene [3 [O - [{L-11 (D1-carbamoyl [3 [carboxypropyl] carbamoylethyl] carbamoylmethyl) -2-dsoxy-αD] glucopydanoside in 320 ml glacial acetic acid is diluted with stirring by the addition of 200 ml of water and the reaction mixture is stirred for 2 hours at a temperature of 50 to 55 [deg.] C. After cooling, the solution is concentrated to dryness. which is always evaporated to give methyl 2-acetamido-3-{- {[11- (D 1 -carbamoyl-3-carboxyphyl) carbamoylethyl] carbamoylmethyl-1 H -deoxy-1 H -glucopyranoside benzyl ester of specific rotation [a] d ²⁰ - + - 64 ¹⁰ + ¹⁰ ° (N, N-diimethylformamide, c = 1.268).

Example 14

A solution of 3.0 g of methyl 2-acetamido-3 [O- [[L-11 [D-carbamoyl-3-carboxy-propyl) carbamoylethyl] carbamoylmethyl] -2-deoxy-6-O-secearyl benzyl ester [ α-D-Glucopyranoside in 100 ml of ethanol and 100 ml of tetrahydrofuran is hydrogenated at atmospheric pressure and at room temperature by addition of 0.6 g-5% palladium on carbon, the catalyst is filtered off and the filtrate is concentrated to dryness under a water pump vacuum. . The crystalline residue is methyl-2-acetamido-3-O - ([L1- (D1-carbamoyl-3-carboxypropyl) carbamoylethyl] carbamoylmethyl} -2-deoxy-6-O-stearoyl-α-D-glucopyranoiside with a specific rotation [a] _D ²⁰ = + + 50 ° + 1 ° (Ν, Ν-dimethylformamide, c = 0.921).

The starting compound is prepared as follows:

To a solution of 3.98 g of methyl 2-acetamido-3-O - {[L1- (D1-carbamoyl-3-carboxypropyl) carbamoylethyl] carbamoylmethyl] -2-deoxy-α-D-glucopyranoside benzyl ester in 40 ml of absolute pyridine was stirred and Exclusion of moisture access dropwise with cooling at 0 to 5 ° over 3 hours a solution of 2.12 g of stearic acid chloride in 20 ml of 1,2-dichloroethane, then the reaction mixture is allowed to stand at room temperature for 18 hours. The reaction mixture was diluted with chloroform, the solution was washed with water, ice-cold 2 N hydrochloric acid solution and again with water, dried with magnesium sulfate and concentrated to dryness. The product, methyl 2-acetamido-3-O - ([L1- (D1-carbamoyl-3-carboxypropyl) carbamoylethyl] carbamoylmethyl] -2-deoxy-6-O-stearoyl-α-D-glucopyranoside benzyl ester, crystallized from ethanol and ether, [α] _D ²⁰ = + 22 ° + 1 ° (chloroform, c = 1.030).

Example 15

A solution of 6.8 g of benzyl-2-acetamido-3-O - ([L-1- (D1,3-dicarbamoylpropyl) carbamoylethyl] carbamoylmethyl) -2-deoxy-α-D-glucopyranoside in 200 ml of 50% aqueous methanol is hydrogenated at room temperature and atmospheric pressure for 60 hours in the presence of 5% palladium on carbon. The catalyst is filtered off and the filtrate is concentrated. The residue was dissolved in 50 ml of water and lyophilized. There was thus obtained 2-Quetaimldo-3-O - [(L1- (D1,3-diicaribamoylpropyl) carbamoylethyl] carbamoylmethyl] -2-deoxy-D-glucose containing 1.24 mol of water as a white powder of [a] D ²⁰ = + 7 ° + ± 1 ° (water, c = 0.514).

The starting compound 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-o-D-glucopyranoside in 100 ml of Ν, Ν-dimethylformamide and 2.77 ml of triethylamine is added 5 ml. L-alanyl-D-glutamic acid diamine hydrochloride and 5.1 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline were allowed to stand at room temperature for 48 hours. After distilling off the solvent, the residue was extracted thoroughly with ether and water and, after drying, recrystallized from a mixture of chloroform and methanol, [ _.alpha. ] _D ^{@ 20} = + + 83 DEG + 1 DEG (.delta., Δ-dimethylformamide, c = 0.531).

A solution of 4 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O - [[L1- (D1,3-dicarbamoylpropyl) carbamoylethyl] carbamoylmethyl] -2-deoxy-α-D-glucopyranoside in 120 ml glacial acid The acetic acid was diluted with 80 ml of water and stirred at 60 ° C for 3 hours. The reaction mixture is cooled, concentrated and the residue is diluted three times in succession by adding 100 ml of water, which is distilled off each time. The benzyl-2-acetamido-3-O - ([L-1,3-dicarbamoylpropyl) carbamoylethyl] carbamoylmethyl} -2-deoxy-α-D-glucopyranoside thus obtained was recrystallized from methanol. Mp 223-225 ° C.

Example 16

A solution of 5.7 g of benzyl 2-acetamido-3-O - {[D1- (D1-carbamoyl-2-carboxypropyl) carbamoylmethyl] carbamoylpropyl] -2-deoxy-α-D-glucopyranoside benzyl ester in 100 ml of glacial acetic acid is hydrogenated in the presence of 5% palladium on carbon at atmospheric pressure and at room temperature. After filtering off the catalyst, the filtrate is concentrated, the remaining 2-acetamido-3-O- (D1 - [(D1-carbamoyl-3-carboxypropyl) carbamoylmethyl] carbamoylpropyl] -2-deoxy-D-glucose is dissolved in 50 ml of water and the solution The substance thus obtained has a specific rotation [α] _D ²⁰ = + 46 ° + 1 ° (water, c = 0.630).

The starting compound used is prepared as follows:

In a mixture of 5.1 ml of 1,2-dichloroethane and 5.1 ml of trifluoroacetic acid, dissolve 5.1 g of N-tert-butyloxycarbonylglycyl-D-isoglutamine benzyl ester and leave the solution to stand for 16 hours. The solution is diluted by adding 100 ml of tetrahydrofuran, the diluted solution is neutralized under external cooling with triethylamine, and a solution of 6.3 g of benzyl-2-acetamido-4,6-O-benzylidene-3-O- (D-carboxypropyl) -2 is added. -deoxy-α-D-glucopyranoside and 1.8 ml of triethylamine in 100 ml of tetrahydrofuran and 3.2 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline. After 24 hours, the reaction mixture was concentrated to dryness and the residue was partitioned between ethyl acetate and water. The organic phase was further washed with 2N ice-cold hydrochloric acid, water, saturated sodium bicarbonate solution and again with water and evaporated. The remaining benzyl 2-acetamido-4,6-O-benzylidene-3-O- [D 1 - [(D 1 -carbamoy 1-3-carboxy propyl) carbamoylmethyl] carbamoylpropyl] -2-deoxy-α D-glucopyranoside benzyl ester was recrystallized from a mixture of tetrahydrofuran and ether. The substance has a specific rotation [α] _ο ²⁰ = + 76 ° + 1 ° (N, N-dimethylformamide, c = 0.457).

A slight hydrolysis in a weakly acidic medium yields the substance in the medium

60% acetic acid Benzyl 2-acetamide benzyl ester-3-O- (D1 - [(D1-carbamoyl-3-carboxypropyl) carbamoylmethyl] carbamoylpropyl) -2-deoxy- and D-glucopyranoside, which is recrystallized from a mixture of methanol and ether. Mp 180-185 ° C [α] _D ²⁰ = + 87 ° + 1 ° (methanol, c = 1.035).

Example 17

4 g of N-tert-butyloxycarbonyl-L-alanyl-D-isoglutamine benzyl ester are dissolved in a mixture of 4 ml of trifluoroacetic acid and 4 ml of 1,2-dichloroethane at room temperature and the air mixture is allowed to stand for 16 hours. . The reaction mixture is then diluted by the addition of 30 ml of 1,2-dichloroethane, neutralized by external cooling with triethylamine, and a solution of 3.7 g of benzyl-2-acetamido-3-O-carboxymethyl-2-deoxy-QD-glucopyranoside with 1 38 ml of triethylamine in 100 ml of tetrahydrofuran. After addition of 2.6 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline, the reaction mixture is allowed to stand at room temperature for 24 hours and then concentrated to dryness. The residue was dissolved in a 9: 1 mixture of chloroform and methanol, and this solution was stirred overnight. washed with water, 2N solution; hydrochloric acid, chilled in ice, water, saturated aqueous sodium bicarbonate solution, and then the solvent is distilled off. The benzyl 2-acetamido-3-O ([11-carbamoyl-3-carboxypropyl) carbamoylethyl] carbamoylmethyl] -2-deoxy-α-D-glucopyranoside benzyl ester was recrystallized from ethanol. Mp 208-212 ° C, [α] _D ²⁰ = + 77 ° ± 7 ° (N, N -dimethylformamide, c = 0.546).

After cleavage of both benzyl residues by hydrogenolysis as described in Example 2, 2-acephainido-3-oxo-1-carbamoyl-3-carboxypropylcarbamoyl-4-ethyl] carbamoylmethyl-2-dodoxy is obtained. -D-glycose. [a] D ^{2 <=} 0 4-Lo ° ± 1 ° (water, c = 0.892).

Example 18

A solution of 3.8 g of benzyl-2-acetamido-3-O - [[L1- (D1,3-dicaboxypyl) -carbamoylethyl] carbamoylmethyl-2-deoxy-o-glucopyranoside dimethyl ester in 100 ml of methanol is treated with The catalyst is filtered off and the filtrate is concentrated to dryness, the residue is dissolved in 70 ml of water, and the solution is lyophilized. acetamido-3-O - [[L1- (D1,3-dicarboxypropyl) carbamoylethyl] carbamoylmethyl] -2-dimethoxy-D-glucose o-specific rotation [α] D ²⁰ = + 23 ° + 1 ° (water) c = 0.814).

D-glutamic acid and 6.95 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dl-zincolizate were then allowed to stand at room temperature for 20 hours. After distilling off the solvent, the residue was dissolved in chloroform and the organic solution was washed with water, 2N ice-cold hydrochloric acid solution, again with water, saturated sodium bicarbonate solution and again with water. After drying over magnesium sulfate, the filtrate is concentrated to dryness. The residue - extracted with hot ethanol - insoluble benzyl-2-acetamide o-4,6-O-benzyl-3-O - {[L-1-

(D1,3-Dicarboxypropyl) carbamozlmethyl-2-deoxy-.alpha.-D-glucopranoside is filtered off and dried; [α] D ²⁰ = + 22 ^p + + Γ (chloroform, c = 1.160).

A solution of 15.6 g of benzyl-2-acetamido-4,6-O-benzidine-3-O - {[L1- (D-1,3-dicarboxypropyl) carbamoylethyl] carbamoylmethyl] dimethylethyl ester | The 2-deoxy-α-D-glucopranoside in 420 ml of glacial acetic acid and 280 ml of water is allowed to warm to -10 DEG C., the reaction mixture is stirred for 4 hours at this temperature, then cooled and concentrated to dryness. 100 ml of water are added to the residue, which is distilled off to dryness, the residue is dissolved in chloroform, washed with water, dried over anhydrous magnesium sulphate and concentrated to dryness. There was thus obtained benzyl-2-acetamide dimethyl-3-O - ([L-11] dimethyl ester).

- (D1, 3-long (-boxpropyl (carbamoylethyl) carbamoylmethyl) -2-deoxz-α1D-glcoprazrazide as a yellowish resin mass, [α] _D ²⁰ = - + 31 ° + 1 ° (chloroform, c = 1,070 ).

Ex - k 1 and -d 1 9

A solution of 6,1 g benzyl-3-O-1 (L-1-

- (D1-Carbamoyl-3-carboxzpropyl) carbamethylmethyl] carbamylmethyl) -2-deoxz-21-propionamido-α-glucopyranoside in -200 ml of a mixture of tetrahydrofuran and water (2: 1) is hydrogenated in the presence of 0.6 g 5% palladium on carbon at atmospheric pressure and room temperature. After the hydrogen uptake is complete, the catalyst is filtered off with suction - and the filtrate is evaporated. 150 ml of water are added to the residue and the reaction mixture is hydrogenated in the presence of 5% palladium on carbon until the consumption of hydrogen ceases. The catalyst is then filtered off and the filtrate is lyophilized. There was thus obtained 3-O - ((L1- (D 11 -carbamoz 13-aaboxylpropyl) carbamoylethyl] acarbamoylmethyl) -2-deoxy-2-propionamido-D-glucose α [α] _D 20 = + 8 ° + 10 ° (water, c = 1.146).

The starting material used can be prepared as follows:

To a solution of 90 g of benzyl-2-acetamido-2-deoxZ-α-D-glucopyranotide in 900 ml of N, N-dimethylformamide was added 0.3 ml of methanesulfonic acid while stirring, eliminating the ingress of moisture and cooling with ice water. solution of 60 ml itopropezzlmethyl ether - in 240 ml N, N-dimethylformamide, reaction

The starting compound used can be prepared as follows:

To a solution of 12.9 g of benzo-2-acetamido-4,6-O-benzzepez-3-O-aarboxymethyl-2-deoxy-α-D-glucorsoroside and 4.0 ml of methylamine in 100 ml Ν, Ν1DisplayZzlform.amide is added

8.1 g of L-alanyl205025 dimethyl ester hydrochloride were stirred at room temperature for 2 hours and basified by addition of triethylamine. The solvent was distilled off, the residue was dissolved in ethyl acetate, this solution was washed with water, dried over magnesium sulfate and the filtrate was concentrated to dryness. The product, i.e. benzyl-2-acetamido-2-deoxy-4,6-O-isopropylidene-α-D-glucopyranoside, is crystallized from ether; mp 136-137 °, [α] _D 20 = + 103 + 1 ° (chloroform, c = 1.125).

In a solution of 225 g of potassium hydroxide in 750 ml of ethanol and 40 ml of distilled water, 52.5 g of benzyl-2-acetamido-2-deoxy-4-bromo-4-oxo-piperidine-α-glucopyranoside are dissolved and the reaction mixture is heated for 4.5 hours. reflux. After cooling, the reaction mixture is concentrated to half its original volume, the concentrate is poured onto ice, extracted with chloroform, the organic phase is washed with water, dried over anhydrous magnesium sulphate and the filtrate is concentrated to dryness. The residue, i.e., benzyl-2-amino-2-deoxy-4,6-O-isopropylidene-α-D-glucopyranoside, is crystallized from ether. The substance has a melting point of 145-146 °, [α] ₀ 2β = + 117 * 04-1 ° (chloroform, c = 1.295).

To a solution of 24.7 g of benzyl-2-amino-2-deoxy-4,6-O-sopropylidene-α-D-glucopyranoside in 192 ml of chloroform is added a solution of 16.0 g of potassium potassium carbonate in 192 ml of distilled water and the reaction mixture was cooled to 0 °. Subsequently, 8.16 g of propionic acid chloride are added with stirring over 20 minutes and the reaction mixture is stirred at this temperature for a further 50 minutes. The organic phase is separated, washed with water, dried over anhydrous magnesium sulphate and the filtrate is concentrated to dryness. The product, i.e. benzyl-2-deoxy-4,6-O-soprorylidene-2-rrorionamido-α-D-glucoryranoside, crystallized from a mixture of ethyl acetate and petroleum ether, m.p. 121-122 ° and [+] _D 20 = + 1120 + 1 ° (chloroform, 0 = 0.977),

To a solution of 9.1 g of benzyl-2-deoxy-4,6-O-isopropylidene-2-rrorionamido-α-D-glucoryranoside in 90 ml of aceeonitrile was added 1.25 g of sodium hydride (Fluka) and the reaction mixture was stirred 2 hours at 40 °. The reaction mixture was then cooled to -5 to -10 ° and added thereto

4.2 ml ethyl bromoacetate. After a further 20 minutes, ethanol (10 ml) was added, the reaction mixture was neutralized with glacial acetic acid and concentrated to dryness. The residue was partitioned between ether and water, the ether solution was washed with water, dried over anhydrous magnesium sulfate, and the filtrate was concentrated. The product, benzyl-3-O-carboxymethyl ethyl-2-deoxy-4,6-O-isoprorylidene-2-propionamido-α-D-glucopyranoside, is crystallized from ether / petroleum ether, mp 94-95 ° C. , [α] D ²⁰ = +. 145 + 1 [deg.] (Chloroform, c = 1.218).

To a solution of 6.8 g of benzyl 3-O-carboxymethyl-2-deoxy-4,6-O-sopropylidene-2-propionamido-α-D-glucopyranoside ethyl ester in 70 ml of methanol is added 22.5 ml of 1N sodium hydroxide solution. After complete ester hydrolysis, 7.5 ml of 1N hydrochloric acid solution are added and the reaction mixture is concentrated to dryness. The product was dissolved in 50 ml of Ν, Ν-dimethylformamide and condensed with 15 mmol of L-alanine-D-isoglutamine benzyl ester with trifluoroacetic acid in the presence of 3.72 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline. . The reaction mixture was concentrated to dryness and the residue was dissolved in chloroform. The solution obtained is washed with water, 2N ice-cold hydrochloric acid solution, water, saturated sodium bicarbonate solution and water, and then the organic solution is concentrated to dryness after drying over anhydrous magnesium sulphate. The product obtained is crystallized from dilute ethanol and then has a melting point of 177-180 °, [α] _D ²⁰ = + 710 + 1 ° (chloroform, 0 = 1.047).

To a solution of benzyl 3-O - {[L-1- (D1-carbamoyl-3-carboxypropyl) carbamyl] benzyl ester, 8,1 g of benzyl-3-O - {[L-1- (D1-carbamoyl-3-carboxypropyl) carbonyl]]. Ethyl-2d-4-ol-4,6-O-isopropyltdene-2-propylamino-α-D-glucopyranoside in 150 ml of methanol is added 15 ml of 1N hydrochloric acid solution and the reaction mixture is allowed to stand. hour from room temperature. Upon addition of 15 ml of 1N sodium hydroxide solution, the reaction mixture was evaporated to dryness and the benzyl 3-O {[L-1- (D1-carbamoyl-3-carboxyrropyl) carbamoylmethyl] carbomoylmethyl] -2-deoxy- benzyl ester obtained. The 2-propionamido-.alpha., .Gamma.-coryranoside is recrystallized from a mixture of methanol and water. After drying, the solid mp 208-210 °, [-jD ^ ² = + 75 ° + ° (Ν, Ν-dimethylformamide, c =. 1120).

Example 2 0

Catalytic hydrogenation of benzyl 3-O- [1,1'-1- (D-carbamoyl-5-carboxypropyl) carbamoylethyl] carbamoylmethyl [-2-caprooylamido-2-deoxy-α-D-glucoryranoside benzyl ester in Ν, Ν- dimethylformamide in the presence of 5% palladium on carbon gave 3-O - ([L-1- (D1-carbamcyl-3-carboxyproryl) carbamethylethyl] carbamoylmethyl] -2-carrinoylamido-2-deoxy-D-glucose white foam, [ _.alpha. ] _D @ 20 = + 11 @ + 1 DEG (water, c = 1.052).

The starting compound used is prepared as follows:

Analogously to Example 1, 16.7 g of capric acid chloride are added to 24.7 g of benzyl-2-amino-2-deoxy-O-isorropylidene-α-Dg-ukoryrancside, and the reaction mixture is likewise treated. . There was thus obtained benzyl-2-capinoylamido-2-deoxy-4,6-O-isopropylidene-α-D-glucoryranostd [.alpha.] D @ 20 = + 81 ++ + 1 DEG (chloroform, c = 1.019).

To a solution of 3.8 g of benzyl-2-caprinoylamido-2-deoxy-4,6-O-sproprene-α-D-glucopyranoside in 40 ml of acetone is added 0.4 g of sodium hydride and the reaction mixture is treated with was stirred at 40 ° for 2 hours. Then the reaction mixture was cooled to -10 °, added. with 1.4 ml of ethyl bromoacetate, and the reaction mixture is stirred for an additional hour at 0 °. Workup as described in Example 19 affords benzyl-3-O-carboxymethyl-2-kaiprinoylamino-4, 6-O-isopropylpyridine-2-deoxy-α-D-glucopyranoside ethyl ester as a tan oil of [ α] D, 2 ° = + (chloroform, c = 1.242).

Analogously to Example 19, the ethyl ester is hydrolyzed, the product is condensed with L-amino-D-isoglutamine benzyl ester with trifluoroacetic acid in the presence of 2-ethoxy-N-sthoxycarbonyl-1,2-dihydroquinoline, and the isopropyl propyl ketal is cleaved by hydrolysis in a slightly acidic medium. . Benzyl 3-O- ([L, Z1- (D-carbamoyl-3-carboxypropyl) carbamoyl-ethyl] carbamoylmethyl] -2-cap [inoylamido-2-deco-α-D-glucopyranoside benzyl ester is obtained. 192 DEG-193 DEG.

Example 2 1

5% solution of benzyl-2-acetamido-2-deoxy-3-O - [[L1- (D1-bis-methylcarbamoylpropyl) carbamoylethyl] carbamoylmethyl-.alpha.-D-glucopyranoside in distilled water / methanol (1: 1) is hydrogenated using 5% palladium on carbon catalyst and the filtrate is evaporated. The product, i.e. 2-acetamido-2-deoxy-3-O - ([Ll- (l, 3-bis-methylcarbamoylpropyl) carbamoylmethyl jkarbamoylmethy 1} -D-glucose. Lyophilized to afford, [a] _D ² 0 = = + 31 ° + 1 ° (water, c = 0.410).

The starting compound can be prepared as follows:

To a solution of 9.1 g of benzyl-2-acetamido-4,6-ol-benzylidene-3-O-carboxymethyl-2-deoxyla-D-glucopyranoside in 100 ml of N, N-dimethylformamide and 2.77 5.6 ml of L-alanyl-D-glutamic acid bis-methylamide hydrochloride and 5.1 g of 2-ethoxy-N-ethoxycarbonylb + dihydroquinoline are added to triethylamine and the reaction mixture is allowed to stand at room temperature for 48 hours. The soap was then distilled off, water was added to the oily residue, the insolubles were filtered off, washed with water and dried. This product was stirred twice more with ether, filtered off and dried. Benzyl-2-acetamido-4,6,10-benzylidene-2-deoxy-3- (O [L-1- (D-1,3-bis-methylcarbamoylpropylcarbamoylethyl) carbamoylmethyl-α-D-glucopyranoside) was obtained.

Slight acid hydrolysis, as described in Example 15, cleaves the benzylidene group and the product, i.e. benzyl-4-acetamido-4-deoxy-3-O - [[L (D1,3-bis)] using 60% acetic acid. (methylcarbamoylpropylcarbamoylethyl) carbamoylmethyl-.alpha.-D-glucopyranoside, m.p. 280 DEG.

Example 2 2

A 5% aqueous solution of benzyl-2-acetamido-3-O - ([L1- (D1-carbamoyl-1-lkabooχpropy-) car26-bamoyl-2-halo-γ-ω-ethyl] carbamoylethyl] -2,2'-deo-α-D-glucopyranoside was added. hydrogenated in the presence of 5% palladium on carbon, after filtration, the filtrate was lyophilized to give 2-acetamido-3-O-11-l- (D1-carbamoyl-3-carboxyphyl) carbamoyl-2-hydroxyoxyethylcarbamoylmethyl] - 2-deo-x-D-glucose [α] D ²⁰ = + 10 + -1 ° (water, c = 1.053).

The starting material can be prepared as follows:

To a solution of 3.7 g of benzyl-2-acetamide-1,3-O-carboxymethyl-2-deoxyla-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 grams of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline The reaction mixture was allowed to react for 20 hours at room temperature, then the solvent was distilled off and the residue was dissolved in a 9: 1 mixture of chloroform and methanol. The solution was washed with water, ice-cold 2N hydrochloric acid, water, saturated sodium bicarbonate solution and water, and the solvent was distilled off after filtration. There was thus obtained benzyl 2-acetamido-3-O- ([L1- (D1-caramoyl-3-carboxypropyl) carbamoyl-2-hydroxyethyl] carbamoylmethyl) -2-deexyl-α-D-glucopyranoside tert-butyl ester . [.alpha.] D @ 20 = + 77 DEG + +1 (methanol, c = 0.946).

Mild acid hydrolysis as described in Example 15 was hydrolyzed · tert give benzyll2 acetamíl-to-3-O- ([Ll- (dl-carbamoyl-3-carboxypropyl) ka ^ι rbamoyll2lhydr oxyethyl] karbamoylmeLhyl 12-deo: γ-α-D-β -opyropyranoside ([α] D = + 65 ° + 1 ° (water, c = 0.947).

Example 2 3

A 5% solution of benzyl-4-acetamide-SO-HL-1- (D1-carbamoyl-3-carboxypropyl) carbamoylbutyl] carbamoylmethyl-2-deoxy-ol D1 glucopyranoside in methanol / water (1: 1) is hydrogenated in the presence of 5 % palladium on carbon; After filtration, the filtrate is concentrated and the residue is dissolved in distilled water and the solution is lyophilized. There was thus obtained 2-acetamido-3-O - ([L1- (D1-carbamoyl-3-carboxypropyl) carbamoylbutyl] carbamoylmethyl] -2-deoxy-D1-glucose. [.Alpha.] D @ ²⁰ = + 12 DEG + 1 DEG (water) , c = 0, 722).

The starting compound can be prepared as follows:

To a solution of 3.7 g of benzyl 2-acetyl-1,3,10-carboxymethyl-2-phenoxy-α-D-glucopyranotide and 1.38 ml of triethylamine in 100 ml of tetrahydrofuran is added 3.28 g of L-norvaline-1-lisoglutamine tert-butyl ester hydrochloride and 2.6 g. The 2-ethoxy-N-leoxycarbonyl-1,2-dihydroquinoline, and the reaction mixture is left at room temperature for 24 hours. Thereafter, the reaction mixture was worked up as in Example 24. This gave butyl-2-acetamido-3O205025 tert-butyl ester.

- {[L-1- (D-1-carbamoyl-3-carboxypropyl) carbamoylbutyl] carbamoylmethyl} -2-deoxy-α-D-glucopyranoside. Mp 202-204 °.

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

Example 24

A solution of 8.3 g of benzyl 1-3-O- ([L1- (D1-carbamoyl-3-carboxypropyl) carbamoylethyl] carbamoylmethyl} -2-deoxy-2-stearoylamido-of-D-glucopyranoside benzyl ester in 200 ml glacial acetic acid ® Hydrogenation is carried out in the presence of 5% palladium on carbon at atmospheric pressure and at room temperature After working up of the reaction mixture, 3-0 - ((L1- (D1-carbamoyl-1H-carboxypropyl) carbamoylethyl] carbamoylmethyl) - is obtained as a white powder. 2-deoxy-2'-stearoylamido-D-glucose [α] _D ²⁰ = + 24 ° + δ (dimethylformamide, c = 0.663).

The starting compound 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 Ν, Ν-dimethylpharmamide was added 0.75 g of sodium hydride and the reaction mixture was stirred for 90 minutes at 40 °. Then the reaction mixture was cooled to -5 °, 2.2 ml of ethyl bromoacetate were added and the reaction mixture was stirred at room temperature for 4 hours. After addition of 10 ml of glacial acetic acid and 400 ml of distilled water, the mixture is extracted with chloroform, the organic phase is washed with water, dried and concentrated to dryness. The benzyl-4,6-O-benzylidene-3-O-carboxyimethyl-2-deoxy-2-stearroylaimido-α-D-glucopyranoside ethyl ester thus obtained is crystallized from ethanol. Mp 151-154 ° [α] _D ²⁰ = H-94 ± 10 ° (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 was added 50.1 ml. 1N sodium hydroxide solution, the reaction mixture was stirred at 50 ° C for 90 minutes, then evaporated. The residue is stirred in ice water, the product is filtered off with suction, washed with water, dried and recrystallized from ethanol. Benzyl-4,6-O-benzylidene-3-O-carboxymethyl-2-deoxy-2-stearoylamido-α-D-glucopyranoside sodium salt, crystallizing with 1 mole of water, melts at 225-242 ° (with decomposition). The substance has a specific rotation [α] ο ²⁰ = 4-45 ° + 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-D-glucopyranoside monohydrate in 120 ml of tetrahydrofuran was added 20 mmol of the benzyl ester salt. Of L-alanine-D-isoglutamine with trifluoroacetic acid and 4.915 g (20 mmol) of 2-ethoxy-N-ethoxycarbonyl-1,2-dichloroquinoline, allowed to stand at room temperature for 20 hours, then concentrated to dryness. The residue was dissolved in chloroform, washed with water, 2N ice-cold hydrochloric acid, water, saturated aqueous sodium bicarbonate solution and again with water, then the organic solution was dried over anhydrous magnesium sulfate and the filtrate was concentrated. The product, ie benzyl ester of 1-4,6-O-benzylidene-3-O - ([L1- (D1-carbamoy 1-3-carboxypropyl) carbamoyl] ethylcarbamoylmethyl (-2-deoxy-2-stearoylamido-α-D-glucopyranoside) , was recrystallized from ethanol, [ _.alpha. ] _D ^{@ 20} = + 62 DEG + 1 DEG (N, N-dimethylformamide, c = 1.148).

Slight hydrolysis of this benzylidene derivative in 65% acetic acid gives benzyl 3-O - ([L1- (D1-carbamoyl-3-carboxypropyl) carbamoylethyl] carbamoylmethyl) -2-deoxy-2-stearoylamido-α-D-glucopyranoside benzyl ester, temperature mp 188-189 °, [α] D ²⁰ = + 59 ° + 1 ° (N, N-dimethylformamide, c = 1,022).

Example 25

A 5% aqueous solution of benzyl-2-acetamido-3-O - ([L1- (D1-carbamoyl-3-carboxypropyl) carbamoyl-2-methylpropyl] carbamoylmethyl] -2-deoxy-? D-glucopyranoside is hydrogenated in the presence of 5% palladium to The catalyst was filtered off and the filtrate was lyophilized to give 2-acetamido-3-O - ([L1- (D1-carbamoyl-3-carboxypropyl) carbamoyl-2-methylpropyl] carbamoylmethyl) -2-deoxy-D-glucose [¢] 0 ^ = + 11¾ (water, c = 1.096).

The starting compound for this process can be prepared as follows:

3.38 g of L-valine-D tert-butyl ester hydrochloride are added to a solution of 3.7 g of benzyl 2-acteamido-3-O-carboxymethyl-2-deoxy-α-D-glucopyranoside and 1.38 ml of triethylamine in 100 ml of tetrahydrofuran. -isoglutamine and 2.6 g

Of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline and the reaction mixture is allowed to stand at room temperature for 24 hours. It is then treated in a similar manner as described in Example 24. This affords benzyl-2-acetamido-3-O - ([L1- (D1-carbamoyl-3-carboxypropyl pcarbamoy 1-2-methylpropyl) carbamoylmethyl] tert-butyl ester | -2-deoxy-α-D-glucopyranoside which, by hydrolysis in a mildly acidic environment, affords benzyl-2-acetamido-3-O- | [L1- (D1-carbamoy 1-3-carboxypropyl) carbamoy 1-2-methylpropyl] carboxy (methyl) -2-deoxy-α-D-glucopyranoside [α] _D ²⁰ = + 76 ° + 1 ° (methanol, c = 1.052).

Example 26

5% solution of benzyl-2-acetamido-3-O - ((L1- (D1-carbamoyl-3-carboxypropylcarbamoyl-3-methylbutyl) carbamoylmethyl) -2-deoxy-α-D-glucopyranoside in methanol / water (1: 1) is hydrogenated in the presence of palladium on carbon and after filtration of the catalyst, the filtrate is evaporated, the residue is dissolved in distilled water, the solution is again filtered, lyophilized to give 2-acetamido-3-O - {[L · -1- ( D-carbamoyl-3-carboxylate of boxypr-propyl) carbamoyl-3-methyl-lbutyl] bamoylmethyJ carboxy-2-deoxy-D-glucose. [?] _D ²⁰ = + 16 ^o + 1 ° (water, c = 1.060 ).

The starting compound used can be prepared as follows:

To a solution of 3.7 g of benzyl-2-acetamido-3-O-carboxymethyl-2-deoxy-α-D-glucopyranoside and 1.58 ml of triethylamine in 100 ml of tetrahydrofuran is added 3.52 g of L-tert-butyl ester hydrochloride. leucine-D-isoglutamine and 2.6 grams of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline. The reaction mixture was allowed to stand for 24 hours at room temperature and worked up as described in Example 24. This afforded the tert-butyl ester. benzyl-2-acetamido-3-O - [[L1- (D1-carbanoyl-3-carboxypropyl-3-methylbutyl) carbamoylmethyl] -2-deoxy-α-D-glucopyranoside, and hydrolyzed in mildly acidic medium to prepare this compound . 2-benzyl-ACEEE IMCD ^ ^ - ^ - C ^ ^j) L - L (dI-carbamoyl-3-carboxy propylamino i) carbamoyl-3-methylbutyl] karbamoylmethylj-2-deoxy-D-glucopyranoside. _[a] 20 = + 73a ± l ° (methanol, c = 0.611).

Example 2 7

A 5% solution of benzyl-2-acetamido-3-O - [[L- (D-carbamoyl-3-carroxypropyl) carbamoylpropyl] carbamoylmethyl] -2-deoxyca-D-glucopyranoside in 50% aqueous methanol is hydrogenated in the presence of 5% palladium on carbon at atmospheric at room temperature. After filtering off the catalyst, the filtrate was evaporated. The residue was dissolved in distilled. water and the solution is lyophilized. There was thus obtained - [2- (D-carbamoyl-3-carboxypiperidylcarbamoyl) propyl] carbamoylmethyl] - [2-dtoxy-D-glucose] - [.alpha.] - [.alpha.] D ²⁰ = + 11 ° + Γ · (water, c = 1.019).

The starting compound can be prepared as follows:

To a solution of 3.7 g of t -benzyl [2- [acttamido-3-carboxymethyl-2-dtoxy-α-Dcg-ukopyranotide] and 1.38 ml of triethylamine in 100 ml of tetrahydrofuran is added 3.24 g of tert-hydrochloride. NaL-Aminobutyryl-D-isoglutamine and 2.6 g of 2-ethoxy-N-ethoxycarbonyl [1,2] dihydroquinoline and after standing at room temperature for 24 hours, the reaction mixture is worked up as described above. in Example 22. The resulting benzyl-2-acetamido-3cO {[L1- (D1 [carbamoyl-3-carboxy] propyl) carbamoylpropyl] carramoylmethyl [-2-deoxy-? D-glucopyranoside tert-butyl ester is then hydrolyzed in slightly acidic. medium to form benzyl-2-acetamido-3-O - {[L- (D 1 - (D 1 - (R) -benzoyl-4-carboxyyroyyl) carbamoylpropyl) carbamoylmethyl] -2-deoxy-α.-D-glucopynganoside . [α] ο ²⁰ = = + 85 · ^ο ° + 1 ° (dimethylformamide, c = 0.725).

Example 28

To the solution - 3.5 g of benzyl 2-acetamidoc3-O - {[Ll- (D-LCL 3 ritckaгramoy lyroyyl) karbamoylethyllkarbamoylmethylj-2-deoxy-4,6-O-itoplΌpyliclen- ₁ 3-D-glukopyranot- In 50 ml of glacial acetic acid, 50 ml of distilled water are added dropwise with stirring at 50 DEG for 15 minutes, and the reaction mixture is stirred at this temperature for one hour. After cooling, 100 ml of distilled water are added and the solvent is removed by distillation in a water pump vacuum. Water (20 ml) was added to the residue and the solution was lyophilized. There was thus obtained benzyl-2-acetamidooc3cO - {[L1- (D1,3-bis-carbamoylpropylcarbamoylethyl) carbamoylmethyl] carbamoylmethyl] -deoxy-.beta.-D-glucopyranoside o - [.alpha.] D @ ²⁰ = -44 DEG. + -. Ν, Ν-dimethylformamide, c = 0.989).

The starting compound used can be prepared as follows:

To a solution of 30 mg of renzylc2acetamoc2-deoxy- [beta] -D-glucopyranoside in 300 ml of N, N-dimethylformamide was added, with stirring, external cooling and exclusion of air humidity, -0.1 ml of methanesulfonic acid. A solution of 20 ml of isopropenyl methyl ether in 60 ml of Ν, Ν-dimethylformamide is then added dropwise over 1 hour, the reaction mixture is stirred for 2 hours at room temperature, basified by addition of triethylamine and the benzylacetate ami'dO'-2 crystallized. deoxy-4,6-O-isoproyylidene-3-D-gl ^, Ucopyranoside from ethyl acetate. M.p. = 194 DEG; [.alpha.] D @ ²⁰ = -104 DEG. + -. 1 DEG (chloroform, c = 0.850).

To a solution of 36.2 g of benzyl 2-acetamido-2-dec oxyc4,6-0 - sopropylidenc _| 3.95 g of sodium hydride are added to the 3-D-glucopyanotide in ~ 400 ml of actethyl ether and 100 ml of Ν, Ν-dimethylformamide and the reaction mixture is stirred for 2 hours at 40 °. Thereafter, the reaction mixture was cooled to -10 DEG C., 17.2 ml of n-butyl bromoacetate was added to the cooled reaction mixture, and the reaction mixture was stirred for 30 minutes while cooling to -0 DEG. After addition of 40 ml of ethanol, the reaction mixture is neutralized with glacial acetic acid, concentrated to dryness, the residue is partitioned between ether and distilled water, the ether solution is washed with water, dried over anhydrous magnesium sulphate and evaporated. The product, i.e. ethyleste.r renzylc2cacetamidoc3-0 ka ^ι гboxymethyl 2cdeoxy-4,6-O-isopropyl-identi- / 3-D-glukoyyгanosidu, SE- recrystallized from ether. Solid, mp 93-94 ° -a [a] D @ ²⁰ = --4 ^с ° ± 1 ° (CHCl, -c = 1 .001).

To a solution of -6.56 g of benzyl-2-acetamido-3-α-cargamoylmethyl-2-deoxy-4,6-O-itoyroyyl-cidene-β-D-glucopyranotide in 70 ml of methanol is added 22.5 ml of 1N sodium hydroxide solution. After ester hydrolysis is complete, 7.5 ml of 1N are added. The solution is concentrated and the reaction mixture is evaporated to dryness. The sodium salt obtained is dissolved in -50 ml of Ν, Ν-dimethylformamide - as a solution - 3.7 g of La-diamide hydrochloride - D-utam acid 205025 selines and 3.72 g of 2-ethoxy-N-ethoxycarbonyl are added. -1,2-dihydroquinoline. The reaction mixture was then allowed to stand at room temperature for 24 hours, concentrated to dryness, dissolved in water and extracted with methylene chloride and twice with n-butanol. The product obtained as the residue after distilling off n-butanol is benzyl-2-acetamido-3-Oi [L1- (D1,3-bis-carbamoylpropylcarbamoylethyl) carbamoylmethyl] -2-deoxy-4,6-O-isopropylidene - / SD-glucopyranoside.

Example 29

To a solution of 5.1 g of benzyl-2-acetamido-3-O - {[L1- (D1,3-bis-carboxypropyl) carbamoylethyl] carbamoylmethyl] -2-deoxy-4,6-O-isopropyl dimethyl ester. 80 ml of distilled water was added dropwise with stirring at 50 DEG and the reaction mixture was stirred at the same temperature for one hour. Evaporation of acetic acid gave benzyl-2-acetamido-3-O- ([L-1-

- (D1,3,3,16-carboxypropylcarbamoylethyl) carbamoylmethyl] -2-deoxy-3-D-glucopyranoside, m.p. 195 DEG-197 DEG, [ _.alpha. ] _D @ ²⁰ = -31 DEG + 1 DEG (methanol, c) = 0.533).

The starting compound used can be prepared as follows:

As in Example 28, 6.56 g of benzyl-2-acetamido-3-O-carboxmethyl-2-deoxy-4,6-O-isopropylidene-3-D-glucopyranoside ethyl ester is hydrolyzed with sodium hydroxide and condensation is carried out with 4.times. 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 distilling off the solvent, the residue was dissolved in chloroform, washed with water, 2N ice-cold hydrochloric acid solution, water, saturated sodium bicarbonate solution and water and, after drying over anhydrous magnesium sulfate, the filtrate was concentrated to dryness. Benzyl-2-acetamido-3-O- [L-1-] dimethyl ester was obtained as a residue.

- (D1,3-bis-carboxypropyl) carbamoylethyl] carbamoylmethyl) -2-deoxy-4,6-O-isopropylidene-3-D-glucopyranoside. [α] D ²⁰ = -81 ° ± 1 ° (chloroform, c = 0.752).

Example 30

5% benzyl 2-acetamido-3-O- [[1- (D1-carbamoylmethylcarbamoyl-3-carboxypropyl) carbamoylethyl] carbamoylmethyl] -2-deoxy-α-D-glucopyranoside benzyl ester in tetrahydrofuran / water (2: 1) is hydrogenated in the presence of 5% palladium on carbon at atmospheric pressure and at room temperature. After uptake of about 50% of the theoretically required amount of hydrogen, the catalyst was filtered off and the filtrate was evaporated. The residue was now dissolved in distilled water and further hydrogenation was performed. After the theoretical amount of hydrogen has been taken up, the catalyst is filtered off again, and the filtrate is freeze-dried. There was thus obtained 2-acetamido-3-O - [[Ll- (Dl karbamoylmeťhylkarbamoyl-3-carboxypropyl Jkarbamoylethyl] carbamoylmethyl] -2-deoxy-D-glucose as a white powder. _[A] - 20 = 4. -H1F + r (water, c = 0.706).

The starting compound can be prepared as follows:

4.8 g of n-tert-butoxycarbonyl-L-alanine-D-glutamine γ-benzyl ester monohydrate are dissolved in a mixture of 5 ml of trifluoroacetic acid and 5 ml of 1,2-dichloroethane and the solution is allowed to stand at room temperature for 16 hours. no access to moisture. The reaction solution was then diluted with 50 ml of tetrahydrofuran, cooled in an ice bath and neutralized with triethylamine. It is then added to the neutralized solution

3.7 g benzyl-2-acetamido-3-carboxymethyl-2- ₍

-deoxy-α-D-glucopyranoside and 1.38 ml of triethylamine in 100 ml of tetrahydrofuran and 2.6 g are added to the obtained solution.

2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline. The reaction mixture was allowed to stand at room temperature for 24 hours and worked up as described in Example 22. This gave benzyl 2-acetamido-3-O - {[L1- (D1-carbamoylmethylcarbamoyl-3-carboxypropyl) carbamoylethyl benzyl ester]. ] carbamoylmethyl | -2-deoxy-α-D-glucopyranoside. Melting point 190-195 ° C.

Example 31

A solution of 5.2 g benzyl-4,6-O-benzylidene-3-O- benzyl ester [L1- (D1-carbamoyl-3-carboxypropyl) carbamoylethyl] carbamoylmethyl) -2-deoxy-2-p-tolylsulfonylamino-α-D-glucopyranoside was hydrogenated in an atmosphere of palladium on charcoal in 120 ml of 70% aqueous acetic acid at atmospheric pressure and at room temperature; After the theoretical amount of hydrogen has been consumed, the catalyst is filtered off and the filtrate is concentrated to dryness. The residue is dissolved in 30 ml of distilled water and freeze-dried. It is obtained in the form of a white powder

3- O- | [L-1- (D-1-Carbamoyl-3-carboxypropyl) carbamoylethyl] carbamoylmethyl-2-deoxy-1'-p-tolylsulphonylamido-D-glucoe.

The starting compound 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 dropwise while cooling to 5 °. solution of 3.1 g of p-toluenesulfonic acid chloride in 30 ml of methylene chloride and the reaction mixture was allowed to stand at room temperature for 1-6 hours.

Then the solution is washed successively with water,

2N sodium hydroxide solution, water, cold 2N hydrochloric acid solution and water, dried over magnesium sulfate and the filtrate concentrated to dryness. The residue is extracted several times with ether, dried to give benzyl 4,6-O-benzylidene-3-O-carboxymethyl-2-deoxy-2-p-tolylsulphonylamino-α-D-glucopyranoside, m.p. 187 °, [α] ^{20 =} η ^o + l + 75 DEG (chloroform, c = 0.855).

To a solution of 5.7 g of benzyl 4,6-O-benzylidentyl 3-O-carboxymethyl-2-deoxy-2-p-tolylsulfonylamino-α-D-glucopyranosulfide with 1.01 ml of triethyl is added 10 mmol of L-alanyl-D benzyl ester salt. isoglutamine with trifluoroacetic acid and 2.5 g of 2-thioxy-N-ethoxycarbonyl-1,2-dihyclruquinoline and the reaction mixture is allowed to stand at room temperature for 24 hours. The reaction mixture is then evaporated to dryness, water is added to the residue, the insoluble materials are filtered off, washed with water and ether and dried in vacuo. Benzyl-4,6-O-benzylidene-3-O-benzyl ester was obtained as a white powder [L-1- (D-1-carbamoyl-3-carboxypropyl) -carbamoyl] ethyl, rhairolinoethyl-2-deexy-2-p-tolylsulfonylamino-α-D-glucopyirosinide.

Example 32

A mixture of 3.2 g of 2-amino-2-deoxy-3-carboxymethyl-D-glucopyranase, 2.5 g of potassium carbonate and 4.5 g of p-nitrophenyl ester of pivalic acid is stirred in 100 ml of dimethylformamide for 15 hours with heating to 100 g. °. After the salts were filtered off, the filtrate was concentrated in an oil pump vacuum and the residue was filtered through IR-120 as an aqueous solution. Shake the eluate with chloroform, concentrate in vacuo to a syrup. Spreading this syrup with ethyl acetate yields a slightly yellowish powder, ie. 2-pivaloylamido-2-deoxy-3-carboxymethyl-glucose pyranose, m.p. 89-93 °; the sodium salt melts at 80 to 90 °.

The starting compound, i.e., 2-amino-2-deoxy-3-carboxymethyl-D-glucopyrrole, is prepared as follows:

A solution of 10.0 g of 2-phenyl-4,5- (3-O-carb) oxyimelhyl-5,6'-sopropylidene-D-glucoiurano-2-oxaline in 1000 ml of a 2.6N hydrochloric acid solution was heated for 4 hours The precipitated benzoic acid was filtered off and the filtrate was extracted three times with ether. The aqueous portion was adjusted to pH = 7 using a pH meter by adding 3N potassium hydroxide solution, clarified with activated carbon, the filtrate was concentrated in vacuo to a volume of about 500 ml, and 3 liters of acetone were added. After cooling with ice, the precipitate formed is filtered off with suction and dried in vacuo at -40 DEG to give 247 g of a yellowish powder which contains about 85% of potassium chloride and 15% of the title compound.

An additional 22 g of a mixture of the title compound and the corresponding lactam is obtained from the mother liquor.

Reaction of 2-pivaloylamido-2-deoxy-3-carboxymethyl-D-glucopyranose with the .alpha.-amide salt of .beta.-benzyl ester. The L-alanyl-D-glutamic acid was prepared analogously to Example 31 using 2-ethoxy-2-ethoxycarbonyl-1,2-dihydroquinoline in a mixture of dimethylformamide and methylene chloride (1). : 1). Hydrogenation in the presence of 5% palladium on carbon gave 2-trimethylacetamido-2-deoxy-3-O- [11- (D1-carbamoyl-3-carboxypropyl) -1-carbba] after lyophilization. moylethyl] carbamoyl-methyl-1-α, β-D-glucose, [α] D 20 = -7 ° (c = 0.6, methanol).

The following compounds are obtained in an analogous manner:

(a) 2-benzoylamino-3-O- {D-1 - [[1- (D-1-carbonyl) -1 H -carboxyyl] -1-carbamoylethyl] carbamoyl-ethyl-2-deoxy-α, 3-glucose having a decomposition temperature of 140 °;

b) 2-benzoylamino-3-O- | DL- [karbamo- Ll-yl-3-carboxypropyl) carbamoyl methylethyl JkarbanioylpiOpyl - ^ - deoxy-.alpha.-D-glukcsa, mp 114-152 °, [п] о ²⁰ = + 17 ° (in methanol);

c) 2-benzoylamino-3-O - {[L1- (D1-carbamoyl-3-cianboxypropyl) -1-carbamoylmethyl] carbamoylmethyl-2-deoxyta, [3'-D-glucose], melting point 175-177 ° (as hydrate);

(d) 2-benzoyl-β-O- (1-Ь- (1D, 3-dicarboxypropyloxycarbonylethyl) carbonylmethyl-2-deynyl-D-glucopyridine OO 6 ', [α] _D 20 = + 25 ° (water) c = 0.997);

e) 2-Benzoylamino-3-O- [1- [1- (1-D-carbamyyl-3-carboxypropyl) carbamyl-2-hydromethyl] carbamoylmethyl-2-deoxy-D-glucopyratratose, m.p. 100 up to 115 °, [α] _D 2 = + 23 ° (water, c = 0.886);

f) 2- (60207100100-3-0- [1-Ь- (1-DNn-propylcarbamoyl-3-carboxypropyl) -carbamoylmethyl] carbamoylmethyl-2-deoxy-D-glucopyraoyl, m.p. 65-140 ° C; _{(a) D} 2> = + 28 ° (water, c = 1.03);

(g) 2-Bromoylamino-3-O-11-methyl-l- (D-tert-butyl-3-carboxypropyl) carbamoylethyl] carbamylmethyl-3-methyl-d-glucose pyranose, m.p. 110-120 ° C, [π] _λ ²⁰ = + 31 ° (water, c = 0.88);

(h) 2-Benzoyl-3-O- (L1- (D1-carbamoyl-3-carbylo -yprypyl) -carbamoylethyl] -carbamoyl-ethyl-2-deoxy-3-ethyl-D-glucosopyranstd, m.p. 215-217 ° C; ²⁰ = -22 [deg.] (Methanol, c = 0.97);

(i) 2-benzoylmethoxy-3-O - [(D1-carbamoyl-3-carboxypropyl) carbamoylmethyl] carbamoylmethyl-2-deoxy-D-glucose, m.p. 115-155 ° C; (a) _D ^{20 =!} + 34 ° (water, c = 0.81);

38

j) 2-benzoylamino-3-O- [11- (D1, 3-diene) m.p. 82 DEG-143 DEG C., ( _.alpha. ) _D @ ²⁰ = + 24 DEG (water, carbampylpropyl) carbamoylethyl] carboxy = 0 , 98).

bamoylmiethyl-2-deoixy-D-glucopyranose,

Claims (2)

OBJECT OF THE INVENTION A process for the production of glucosamine derivatives of the general formula (I) in which: X a carbonyl group or a sulfonyl group, R is C 1 -C 18 alkyl or phenyl, R 1 is hydrogen or (C 1 -C 5) -alkyl or benzyl, R 2 is hydrogen or C 1 -C 5 alkyl, R 1 and R 6 are hydrogen or an acyl radical having 2 to 18 carbon atoms, R 7 is hydrogen, C 1 -C 5 alkyl, hydroxymethyl or phenyl, Re optionally an esterified or amidated carboxyl group a R9 is an optionally esterified or amidated carboxyl group, provided that the alkyl moiety of R is more than 1 carbon atom when X is carbonyl and R2 is methyl, or when X is carbonyl, R2 is hydrogen and Ra is and R 6 represents a carboxyl group, as well as their pharmaceutically usable acid addition salts, characterized in that the compound of formula III, COOH (III) in which X, R, and R2 are as defined above, and R 10, R ⁴⁰ and R 6 ^Q have the meanings given for the radicals R 1, R 4 and R 6, respectively, or are a readily cleavable capping group, or a derivative of this compound is condensed with a compound of formula IV, R7 ⁰ Re “ H2N — CH — CONH — CH (CH2) z — Ro (IV) wherein 7 ^0, Re and R9 ⁰ have the meanings of the symbols R7, Re and R9, with the carboxylic acid and, optionally, free hydroxy groups present in these radicals are protected by easily cleavable protective groups and optionally present protective groups are cleaved, whereupon the optionally obtained the salts are converted into aqueous compounds or the resulting salts are converted into their physiologically acceptable acid addition salts. 2. A process according to claim 1, wherein the acids of the formula III in the form of an acid anhydride or a reactive ester are condensed with an amino compound of the formula IV or an acid of formula III with a compound of the formula IV.