IE44190B1 - Glucosamine derivatives and a process for their manufacture - Google Patents

Abstract

Glucosamine derivatives of the formula (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 invention relates to new glucosamine derivatives, especially glucosamino-3-alkanoyl-dipeptides. The invention provides glucosamine derivatives of the general formula CONH - CH(CH9), - R„ (D) Z 3 X denotes a carbonyl or sulphonyl substituted alkvl radical or an ίο J.5 and the salts thereof in which group, R denotes an optionally optionally substituted aryl radical and, if X is the carbonyl group, also denotes an alkoxy or benzyloxy radical, Rj denotes hydrogen, alkyl or an optionally substituted benzyl radical, R2 denotes hydrogen or lower alkyl, R^ and Rg denote hydrogen, alkyl or an optionally substituted benzyl or an acyl radical, Ry denotes hydrogen, alkyl, hydroxymethyl, mercaptomethyl or phenyl, Rg denotes an optionally esterified carboxyl group or amidised carboxyl group which is Optionally substituted, and Rg denotes an optionally esterified carboxyl group or amidised carboxyl group which is optionally substituted, with the proviso (a) that the optionally substituted alkyl radical R has more than 1 carbon atom if (1) X denotes the carbonyl group and the radical R2 denotes methyl, or (2), if X denotes the carbonyl group, the radical R2.represents hydrogen and Rg and Rg each represent a carboxyl group, and also with the proviso (b) that where R2 and Ry are other,than hydrogen, the side-chain attached to oxygen in the 3-position of the glucosamine residue has the DLD configuration, d 4 ί. 9 Ο and a proceas for their manufacture.

Union!; spe.yficolly staled, residues, radicals or compounds modified by ihe term lov/er in the text which follows contain up to 7 and preferably up to 4 carbon atoms, Alkyl is, in particular, lower alkyl, for example isopropyl, straight-chain or branched butyl, pentyl, hexyl or heptyl, which are bended in any desired position, and above all methyl; ethyl or n-propyl.

Possible substituents of the optionally substituted alkyl group are free or functionally modified hydroxyl or mercapto groups, such ns etherified or esterified hydroxyl or mercapto groups, for example lower alkoxy or lower alkylmereapto groups, or halogen atoms or free or functionally modified carboxyl groups, such as carbo-lower alkoxy or carbamoyl groups. The substituted alkyl radical, such as a lower alkyl radical, can carry one, two or more identical or different substituents, especially free hydroxyl groups or halogen atoms.

Aryl radicals are, in particular, monocyclic aryl radicals and also bicyclic aryl radicals, especially phenyl, but also naphthyl- They can optionally be monosubstituted, disubstituted or pclysubstituted, for example by lower alkyl groups, free, etherified or esterified hydroxyl, for example lower alkoxy or lower alkylenedioxy, or halogen atoms and/or trifluoromethyl groups.

If the radical X is a carbonyl group, R can also represent an alkoxy radical, especially a lower alkoxy radical, such as the inethoxy or ethoxy radical, or the benzyloxy radical· - 3 44180 The grouping -NH-X-R then forms the radical of an. ester of carbamio acid.

Optionally substituted benzyl radicals are, in particular, those benzyl radicals which are optionally monosubstituted, disubstituted or polysubstituted in the aromatic nucleus, for example by lower alkyl, free, etherified or esterified hydroxyl or mercapto groups, for example lower alkoxy or lower alkylenedioxy groups, as well as lower alkylmercapto or trifluoromethyl groups and/or halogen atoms.

An optionally esterified or amidised carboxyl group is, preferably, the carboxyl group itself or a carboxyl group esterified with a lower alkanol, such as methanol or ethanol, or the carbamoyl group which, on the nitrogen atom, is unsubstituted or1 monosubstituted or disubstituted by alkyl, especially lower alkyl, aryl, preferably phenyl, or aralkyl, such as benzyl.

The carbamoyl group can, however, also carry an alkylidene radical, such as a butylidene or pentylidene radical.

The carbamoyl group Rg can also carry a carbamoyl-methyl group on the nitrogen atom.

Acyl is, in particular, an acyl radical of an organic acid, especially of an organic carboxylic acid. Thus, acyl is in particular alkanoyl, preferably with 2-18 carbon atoms, such as acetyl or propionyl, or also aroyl, such as naphthoyl1-, naphthoyl-2 and especially benzoyl^ or benzoyl or naphthoyl substituted by halogen, lower alkyl, lower alkoxy, trifluoromethyl, hydroxyl or lower alkanoyloxy, - 4 4418ο as well as carbamoyl, such as unsubstituted carbamoyl, lower alkyl-carbamoyl or aryl-carbamoyl, such as methylcarbamoy1 or phenyl-carbamoyl.

As a substituent of the abovementioned radicals, lower alkyl is preferably methyl or ethyl but may be also n-propyl, isopropyl or straight-chain or branched butyl.

As a substituent of the abovementioned radicals, lower alkoxy is, in particular, methoxy or ethoxy and also n-propoxy, isopropoxy, n-butoxy or isobutoxy.

As a substituent of the abovementioned radicals, lower alkylmercapto is above all methylmercapto or ethylmercapto, but also n-propy1-mercapto or isopropy1-mercapto.

Halogen is, for example, fluorine, chlorine or bromine. Carbo-lower alkoxy groups which may be mentioned in particular are carbomethoxy or carboethoxy, but also carbo-npropoxy or carbo-isopropoxy groups, and carbamoyl groups which may be mentioned in particular are the carbamoyl group itself.

In the abovementioned compounds, in which Rj denotes an alkyl radical, the R2-acetamide radical which is linked to the oxygen atom in the 3-position of the glucosamine radical is optically active, that is to say it is in the D-form. If Ry does not represent hydrogen, the Ry-acetamide radical is in the L-form.

Depending on the nature of their substituents, the present new compounds are neutral, acid or basic compounds. If excess acid groups are present, they form salts with bases, such as ammonium salts or salts with alkali metals or alkaline earth metals, for example sodium, potassium, calcium or magnesium. If, however, excess basic groups are present, they form acid addition salts.

Acid, addition salts are, in particular, non-toxic acid addition salts which can be used pharmaceutically, such as those with inorganic acids, for example hydrochloric acid, hydrobromic acid, nitric acid, sulphuric acid or phosphoric acid, or with organic acids, such as organic carboxylic acids, for example acetic acid, propionic acid, glycollic ac.id, succinic acid, maleic acid, hydroxymaleic acid, methylmaleic .acid, fumaric acid, malic acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-aminosalicylic acid, 2-phenoxybenzoic acid, 2-acetoxy-benzoic acid, embonic acid, nicotinic acid or isonicotinic acid, or organic sulphonic acids, for example methanesulphonic acid, ethanesulphonic acid, 2-hydroxy-ethanesulphonic acid, ethane-1,2disulphonic acid, benzenesulphonic acid, p-toluene-sulphonio acid or naphthalene-2-sulphonic acid, and also other acid addi25 tion salts which can be used, for example, as intermediate products, for example for purifying the free compounds or in the manufacture of other salts and can also be used for - 6 44180 churacteristion, for example, those with picric acid, picroionic acic., fl-avianic acid. phosphotungstic acid, phesphomolybdic acid, chloroplatinic acid, Roiuecke's aeid or perchloric acid.

The compounds of the present invention display valuable pharmacological properties, in particular a marked immunityboosting action. This can be deraonsti-ated with the aid of the test arrangement described below: 1. Boosting of the cellular irmraity in vivo: increase in the delayed-type hypersensitivity to .ovalbumin in i.nin-a. uirs Pirbright guinea pigs are immunised cn day 0 with lo rag of ovalbumin in complete Freund’s adjuvant by injecting 0.1 ml of an antigen-adjuvant mixture into each of the two back paws. vzeeks’ later skin reactions are produced by intracutanaous injection of 100 pg of ovalbumin in 0.1 ml of buffered physiological sodium chloride solution and quantified on the basis of the reaction volume calculated 24 hours later with the aid of the surface area of the erythema and of the increase in the thickness of the skin. The antigen-specific increase in the reaction volume which is observed after 24 hours (delayed-type reaction) is taken as a measure cf cell-transmitted immunity. Ovalbumin is too weak an immunogen to induce a delayed-typs reaction on its own or in a water-in-oil emulsion with incomplete Freund's adjuvant (10 parts by weight of ovalbumin solution in 0.9% by weight of NaCl mixed with 8.5 parts by weight of Bayol 1' and 1.5 parts by weight of Arlacel I Trade Mark), A), but, for effective immunisation, has to be administered in complete adjuvant, to which Mycobacteria are added (5 ng of - 7 _ 4419® M butyricum, which have been destroyed and lyophilised, per 10 ml of Bayol r/Arlacel A). In order to demonstrate the immunity-boosting action of the test substances, the latter can now be mixed, in doses of 10 to 100 ug, with the antigen/oil mixture in place of the Mycobacteria* The glucosamine peptides according to the invention are able to imitate the effect of the Mycobacteria in the test arrangement described and quantitatively to surpass this effect.

A significant boosting of the delayed-type reactivity against ovalbumin can also be achieved when compounds of the type described are not incorporated into the antigen/oil mixture but are administered subcutaneously, in sodium chloride solution, in doses of 10 to 100 jig per animal for several days after immunisation (for example on day 0, 1, 2, 5, 6 and 7).

This shows that compounds of the type described are able considerably to increase cellular immunity, both .as a mixture with the antigen itself (adjuvant effect in the narrower sense) and when supplied at times and places which differ from those of the antigen injection (systemic immunity-boosting). 2. Boosting of the humoral immunity in vivo; increase in the production of antibodies against bovine serum albumin (BSA) in mice. .

KMRI mice are immunised by intraperitoneal (i.p.) injection of 10 gg of precipitate-free BSA on day 0. Serum samples are taken S, 15 and 29 days later and tests are carried out, using a passive haemagglutination technique,to determine their content of anti-BSA antibodies. In the dosage used, soluble - 8 4418ο BSA is suh-immunogy.-nlo for the receiving animals, hhat is to say it is not able tc initiate eny production of antibodies or is able to initial·. -/:)1:/ a very si5·,-lit production of antibodies. Additional treaLment of the mice with certain immuni'ty-boosting substances before or after the admin::stration of the antigen leads to 0. rise in the antibody til-·? in the serum. The effect of the treatment is expressed by the score value, that is tc say by the sum of the log., titre differences on the three days when blood .is sampled, which is reached. lo On intraperitoneal or subcutaneous (s.c.) administration of 100-./00 mg/kg/animal on five consecutive days (day 0 to 4) after immunisation with BSA. compounds of the present invention aro able significantly to increase the production of antibodies against. BSA.

The immunity-stimulating effect of the said compounds, in contrast to that of other bacterial immuuoleptic agents (for example LPS from 3. coli), is dependent on the antigen; injection of the new compounds results in an increase in the antiBSA titre only in ESA-immunised mice, but not in mice which have not been .immunised. It is worthy cf note that the s.c. administration of the said compounds is equally as effective as the i.p. administration, that is to say tbe immunity-boosting action which is observed in systemic and does not depend on the stimulant having to be administered by the same route as the antigen or as a mixture with the antigen, as is the case with classical adjuvants.

The experiments iXhatrated show that compounds of the type - 9 441®° described are also able specifically to increase the humoral Immunity, that they improve the immunological response and that their Immunity-boosting effects are based on a systemic activation of the immune system. . 3- Boosting of the humoral immunity in vitro: T-cellreplacement effect in the antibody response of mice splenocytes to sheep erythrocytes (SE)..

In many cases, lymphocytes (T-cells) emanating from the thymus are necessary for the induction of an antibody response.

These cells co-operste with the precursors of antibody-forming lymphocytes (B-cells) and help them to react to stimulation by so-called T-dependent antigens by proliferation, differentiation and synthesis of antibodies. Suspensions of splenocytes from congenitally athymic nu/nu mice do not contain any func15 tional ϊ-cells and, for example in vitro in the presence cf SE, are not able to form any anti-SE antibodies. The compounds of the present invention are, surprisingly, able functionally to replace T-cells in such cultures and to make an antibody response to SE possible. The addition of-ftiese substances to nu/nu splenocyte cultures in the presence of SE leads, within 4 days, to a considerable increase in the number of antibodyforming cells. The findings show that the said compounds are able to increase the humoral antibody formation in vitro and to compensate a defect in the ΐ-cell system. 4, Selective mitogeneity for B-cells: proliferationpromoting effect in B-lymphocyte cultures.,.

Suspensions of highly enriched E-lymphocytes 441S0 (lymph node ceils £::0::1 ο-.>£·„:ώ tally athymic nu/nu mice) and substantially pare imuatura and mature T-lymphocytes (thymocytes and, respectively, cortisone-resistant thymocytes, that is tc say thywocyten which persist 48 hours aftsr a cortisone injection, from Baib/c mice) are incubated for· three days in the presence of the test substances. The incoa^poration of H-'~ thymidine in the lymphocytes during the final 18 hours of tne culture period is taken ss a measure of the proliferation acta•vity.

The compounds of the invention are mitogenic for B‘lymphocytes (that is to say for the precursors of the antibodyforming colls) but not for T-lymphocytes.

They are thus able to stimulate the proliferation of lymphocytes which are involved in the humoral immune response.

Although compounds of the type described already di..-.— play their* boosting action in guinea pigs after, fox· example, an individual dose of 0.0(5 mg/kg administered subcutaneously and in mice aflrr the subcutaneous administration of 5 times 10 mg/kg, no toxic effects are observed in mice even when 5 times 300 mg/kg are administered intraperitoneally. The said substances therefore possess an excellent therapeutic range.

The compounds according to the invention are able when mixed with an antigen, to increase the immunogeneity of the latter and also, on systemic administration, to increase! the immunological reactivity cf the treated organism. The said substances are able to promote - 11 441S0 both the cellular and. the humoral immunity and to activate the lymphocytes which are responsible for the formation of antibodies.

The new compounds can thus be used, as adjuvants mixed with vaccines, to improve the success rate of the vaccination and to improve the protection imparted by humoral antibodies and/or cellulax· immunity against infection by bacterial, viral or parasitic pathogens.

Finally, the compounds described are suitable, as a mixture with very diverse antigens, as adjuvants fox· the experimental and industrial production of antisera for therapy and diagnosis and for inducing iramunologically activated lymphocyte populations for cell transfer processes· Moreover, the new compounds can be used, even without the addition of an antigen at the same time, to promote immune reactions which are already taking place subliminally in humans and animals. Accordingly, the compounds are especially suitable for stimulating the body’s own resistance, for example in the case of chronic and acute infections or in the case of selective (antigen-specific) immunological defects, and also in the case of general (that is to say not antigen-specific) immunological defective states v/hich are congenital ox· acquired, such as arise in old age, in the course of severe primary diseases and, especially , after therapy with ionising rays or with hormones having an immune-suppressive action. ' The said substances can thus be administered, preferably also in combination with anti-infectious antibiotics, chemotherapeutic agents or 443.90 other· courses c.f ί. ,-, ίιί·::;ιί, in, order to '.ouritor.-rot c;, :.:9 damage. Finally, the substances described are al >'· sults;·': ·:for the general prophylaxis of infectious diseases in human? snd animals.

The invention relates in particular to compounds of the formula (I) in which X denotes a carbonyl radical and R denotes a lower alkyl radical which is optionally substituted hy hydi-S'-tyl or carboxyl groups or denotes a phenyl radical whioh is optionally substituted by lower alkyl, lower alkoxy, triiluoromethyl cn' halogen, and their salts.

Compounds in which Rg represents hydrogen and their salts are also particularly valuable, Compounds to be singled out are, in particular, compounds of the general formula (II.) (C1I2)2 - K, in which R denotes lower alkyl or? phenyl, R^ denotes hydro Ο X lower alkyl, Rg denotes hydrogen or methyl, Ry denotes hydrogen. - 15 ~ 4419® lower alkyl or hydroxymethyl, Rg denotes carbamoyl and Rg denotes carboxyl, with the proviso that the lower alkyl radical R contains more than 1 carbon atan if Rj denotes methyl, and their salts.

Compounds to be mentioned above all are those of the formula II in which Rj_ denotes hydrogen, and R? denotes hydrogen, methyl or hydroxymethyl.

Other categories of ccnpound to be singled out are:1) compounds of formula I, in which and R^ denote hydrogen, R denotes phenyl, R? denotes methyl, and Rg and Rg denote carboxyl, carbanethoxy, carbamoyl, or N-methyl-carbamoyl; 2) . compounds of formula II, in which R^ and Rj denote hydrogen, R denotes nethyl or phenyl, R? denotes hydroxymethyl, Rg is carbamoyl, and Rg is carboxyl; 3) ccnpounds of formula II, in which R^ and Rj denote hydrogen, R denotes phenyl, R? is nethyl, Rg is carboxyl, and Rg denotes N-propyl-carbamoyl or carbampyimethylcarbampyl; 4) ccnpounds of formula I, in which R^ and R2 represent hydrogen, R and R? denote methyl, and Rg and Rg denote carbanethoxy, carbamoyl, or N-metbylcarbanqyl; 5} ccnpounds of formula 1, in which R denotes phenyl, R^ denotes ethyl, Rj denotes hydrogen, R? is methyl, Rg is carbamoyl and Rg is carboxyl, or Rg and Rg denote carbanethoxy or N-methylcarbampyl; 6) ccnpounds of formula I, in which R and R? denote methyl, R^ is benzyl, Sj is hydrogen, and Rg and Rg represent carbanethoxy or carbamoyl.

Hie new ccnpounds may be obtained when a compound of the formula in which X, R and Rg possess the abovanentioned meaning and R^°, R4° and Rg° represent the radicals R^, R4 and Rg respectively or represent a protective group is condensed with a compound of the formula - 14 44190 j<7° |e° (iv) UN - CH - CONH - CH(CH2)? - Η?° in which ΗγRQ° and Rg° possess the meaning of R.·,, Rg and Rg, with ths proviso that carboxy1 groups and, if desired, free hydroxyl groups present in these radicals are protected by protective groups and protective groups which are present are split off.

The condensation reaction is carried out, for example, by reacting the compound III, in the form of the activated carboxylic acid, with the amino compound IV or by reacting the acid III with the compound IV, in which the amino group in present in an activated form. The activated carboxyl group can bo for example, an acid anhydride, preferably a mixed acid anhydride, such as an acid azide, an acid amide, such as an imiuazolide, an isoxazolide or an activated ester. Activated esters which may be mentioned in particular are: cyanomethyl esters, carboxymsthyl esters, p-nitrophenylthio esters, p-nitrophenyl esters, 2,4,5trichlorophenyl esters, pentachlorophenyl esters, N-hydroxysuccinimi.de esters, K-hydroxypthalimide esters, 8~hydroxyquino~ line esters, 2-hydi'oxy-l,2“dihydro-l-carboethoxy-Quinoline esters, K-hydroxypiperidino esters or enol esters which are obtained with M-othyl~5“phenyl-isoxazol.ium 3' -sulphonate. Activated esters can also optionally be obtained with a carbodiimide with tho addition of N-hydroxysuccinimide cr a 1-hydroxybenzotriazole cr 3--hydroxy-4-oxo-3,4-dihydro-benzo[d]-l,2,3-triazine which is - 15 44190 unsubstituted or substituted by, for example, halogen, methyl or methoxy.

The amino group is activated by, for example, reaction with a phosphite-amide.

Amongst the methods cowrising · a reaction with activated esters, those with K-ethyl-5-phenyl-isoxazolium 3‘-sulphonate (Woodward reagent K) or 2--ethoxy-l,2-dihydro-l-carboethoxyquinoline or carbodiimide are to be mentioned in particular.

Protective groups which can be split off easily are those which are known from peptide chemistry and sugar chemistry. Protective groups which should be mentioned for carboxyl groups are, in particular, tertiary butyl, benzyl or benzhydryl and for hydroxyl groups are, in particular, acyl radicals, for example lower alkanoyl radicals, such as acetyl, aroyi radicals such as benzoyl, and, especially radicals derived from carbonic acid, such as benzyloxycarbonyl or lower alkoxycarbonyl, or alkyl, especially tert,-butyl, benzyl or tetrahydropyranyl which are optionally substituted by nitro, lower alkoxy or halogen, or optionally substituted alkylidene radicals which link the oxygen atoms in the 4-position and 6-position. Such alkylidene radicals are, in particular, a lower alkylidene radical, above all the ethylidene, isopropylidene or propylidene radical, or also a benzylidene radical which is optionally substituted and preferably substituted in the p-position.

These protected groups can be split off in a conventional manner. Thus, they can be removed hydrogenolytically,· for example with hydrogen in the presence of a noble - 16 4419ο metal hydrogenation catalyst, such as a palladium or platinum catalyst, or by acid hydrolysis.

The starting materials used are known or can be manufactured in a conventional manner. Thus, compounds , of the formula III can be obtained, for example, by reacting the corresponding sugar, whioh is unsubstituted in the ^-position, with a halogeno-Rg-acetic acid, in which Rg has the abovementione: meaning, and its esters in the presence of a strong base. In these compounds, halogen is preferably bromine or especially chlorine.

Another procedure for the manufacture of the new glucosamine derivatives is to condense a compound of the formula V CONH - CH - COOH in which R, R^°, Rg, R^°, Rg° and Ry° have the abovementioned meaning, with a compound of the formula V ' . . HZN - CH - (CH2J2 - R9 (VI) - 17 44190 in which Rg° and R^0 have the abovementioned meaning, with the proviso that carboxyl groups and, if desired, free hydroxyl groups present in the radicals R?0, Rg° and Rg° are protected by protective groups which can. be split off easily, and to split off any protective groups which may be present.

The condensation reaction is carried out, for example, by reacting the compound V, in the form of the activated carboxylic acid, with the amino compound VI, or by reacting the acid V with the compound VI, in which the amino groups are in an· 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. These can be, in particular, the acid anhydrides, amides or esters mentioned above. The amino group is activated, for example, by reaction with a phosphite-amide.

The protective groups which can be split off easily also correspond to those already mentioned above. They can be split off in a conventional manner; for example .hydrogenolytically, for example with hydrogen in the presence of '20 . a noble metal hydrogenation catalyst, such as a palladium or platinum catalyst, or ..by acid hydrolysis.

The starting materials can be obtained in a conventional manner. Thus, it is possible, for example, to react corresponding sugars which are unsubstituted in the 325 position with a halogeno-R2-acetamido-acetic acid substituted by Ry> at 'the alpha carbon atom or to react a compound of the formula III in the manner indicated above with a similarly substituted amino-acetic acid in which the carboxyl group - 18 4 418ο is protected and to split off the protect!va group.

A further process method for introducing the side chai located in the 3-position of the sugar residue is to react a compound in which X, R, R^°, RZl° and R^0 have the abovementioned meaning end any hydroxyl, groups which may bo contained therein are pro tectcd by a protective group, with a coinpound of the formula - CH - COSKCH - COHH ~ CH - (CI12)2 - Rg (VIII) in which Z represents areactively esterified hydroxyl group and Ry°, R^° and R^° have the abovementioned meaning, and to split off any protective groups which may be present.

A reaclvely esterified hydroxyl group is, in particular 15 a hydroxyl group esterified with a strong inorganic or organic aeid and# especially,a hydroxyl group which is esterified with hydrogen halide acids, such as hydrochloric acid., hydrobromic - 19 ~ 44190 acid ο/? hydriodie acid.

The protective groups which can be split off easily correspond to those already mentioned above. They can be split off in a conventional manner, for example hydrogenolytically, for example with hydrogen in the presence of a noble metal -hydrogenation catalyst, suoh as a palladium or platinum catalyst, or by acid hydrolysis.

The starting materials used in this process variant are known.

However, the compound may also be obtained when the oxasoline ring and the dioxolane formula IX ring m a compound oi the CH - (CU2)2 - R9 IX possess the abovementioned in which R, Rg, Ry°, Rg° and R^° meaning and R^ is an alkylidene or cycloalkylidene group, is split open under acid conditions with formation of the glucosamine ring and any protective groups which may be present are split off and if the radical R-CO- has been removed, the X-R radical is .introduced into the amino group in the 2-position of the sugar molecule if this group has been liberated.

In this compound, alkylidene is, in particular, lover alkylidene, such as isopropylidene, and cycloalkylidene is preferably cyclopentylidene or cyclohexylidene.

This splitting is also carried out in a conventional 5 Manner, for example with an acid ion exchanger, especially those containing sulphonic acid groupe, such as Amberlite (Registered Trade Mark) IR-120 (a styrene reein with highly acid sulpho groupe) or Dcwex . — :(tode Mark) 50 (polystyreneeulphcnic acids) or a strong inorganic or organic acid, such as hydrochloric acid., hydrobromic acid, sul10 pburic acid or a sulphonic acid, for example methanesulphonic acid, or a phenylsulphonic acid which is optionally substituted in tlie aromatic ring, such as p-tolusnesulphonic acid, or trifluoroacetic acid. If the reaction is carried out in the presence of water a free hydroxyl group is obtained in the 1-positicn; ifj on the Other hand, the reaction is carried out in the presence of an alcohol of the formula 110-R^ in whic’n R^ represents an optionally substituted alkyl radical, the 1-0-R^ compound is obtained. If one of the carboxyl groups Ηθ and/or Rg is also esterified with an alcohol, especially a lower alkanol, this can be saponified with aqueous acid, especially at elevated temperature.

However, it is also possible for the amino group in the 2-position of tlie sugar molecule to be liberated during this splitting. In this case, the X-R radical has to bo intro25 duced subsequently. This is carried out An the customary manner by acylation or sulphonylation.

In the resulting compounds it· is possible subsequently to sp3.it off protective groups on the peptide radical, for example by hydrogenolysis, such as, for example, with catalytically activated hydrogen·, or by hydrolysis.

The starting materials used can be obtained, for example, . when the Rg-acetamidopeptide radical is introduced, in one or several stages, into a corresponding ozaz&line which has a free hydroxyl group in the 3-position of the sugar residue..

The resulting compounds can be converted into their salts in a conventional manner, for example by reac10 ting resulting acid compounds with alkali metal hydroxides or . alkaline earth metal hydroxides or by reacting resulting basic compounds with acids.

The processes described above are carried out by conventional - methods which are in themselves known, in the absence or, preferably, in the presence of diluents or solvents, if necessary vzith cooling or warming, under elevated pressure and/or in an inert gas atmosphere, such as a nitrogen atmosphere.

Taking into account all of the substituents present in the molecule, particularly gentle reaction conditions, such as short reaction times, the use of mild acid or basic agents in low concentration, stoichiometric ratios and the choice of suitable catalysts, solvents and temperature and/or pressure conditions, must be employed if necessary, especially in the presence of O-acyl radicals which can be hydrolysed easily.

The invention also relates to those embodiments of the process according to which -52 — a starting material is used in the form of a reactive derivative or salt. The starting materials preferably used are those Which, according to the process, lead to those compounds described above as being particularly valuable.

The present invention also relates to pharmaceutical formulations Which contain compounds of the formula I together with a pharmaceutical excipient. The formulations may also contain a vaccine. The pharmaceutical formulations according to the invention are those for enteral, such as oral or rectal, and parenteral administration to warm-blooded animals and which contain the pharmacological active compound on its own or together with an excipient which can be used pharmaceutically. The dosage of the active compound depends on the species of warm-blooded animal, on the age and on the individual state and on the mode of administration.

The new pharmaceutical formulations may contain from 10% to 95% by weight, and preferably from 20% to 90% by weight, of the active compound. Pharmaceutical formulations according to the invention can, for example, be in a dosage unit form, such as dragees, tablets, capsules, suppositories or ampoules.

The pharmaceutical formulations of the present invention are manufactured in a conventional manner, for example by swans of conventional mixing, granulating, dragee making, dissolving or lyophilising processes. Apart from the types of administration mentioned herein pharmaceutical 441»° formulations especially for oral use can also be obtained by combining the active compound with solid excipients, optionally granulating a resulting mixture and processing the mixture or granules, after adding suitable auxiliaries if desired or neces5 sary, to give tablets or dragee cores.

Suitable excipients are, in particular, fillers, such as sugars, for example lactose or sucrose, mannitol or sorbitol, cellulose formulations and/or calcium phosphates, for example tricalcium phosphate or calcium hydrogen phosphate, as well as binders, such as starch pastes using, for example, maize starch, wheat starch, rice starch or potato starch, gelatin, tragacanth, methylcellulose, hydroxypropylmethylcellulose, sodium oarboxymethylcellulose and/or polyvinylpyrrolidone, and/or, if desired, disintegrating agents, such as the abovementioned starches, and also carboxymethyl-starch, crosslinked polyvinylpyrrolidone, agar or alginic acid or a salt thereof, such as sodium alginate. Auxiliaries are, especially -, flow-regulating agents and lubricants, for example silica, talc, stearic acid or salts thereof, such as magnesium stearate or calcium stearate, and/or poly20 ethylene glycol. Dragee cores are provided with suitable coatings which, if desired, are resistant to gastric juices, and for this purpose. Inter alia, concentrated sugar solutions, which optionally contain gum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and/or titanium dioxide, lacquer solutions in suitable organic solvents or solvent mixtures or, in order to produce coatings resistant to gastric juices, solutions of suitable cellulose formulations, such as acetylcellulose phthalate or & hydroxypropylmethylcellulcse phthalate, are used. Dyestuffs or pigments can be added to the tablets or dragee coatings, for example for identification or' .i.n order to characterise different doses of the active compound.

The examples which follow illustrate the invention described above; they are not, however, intended to restrict the scope of the invention in ary way. Temperatures are given in degrees centigrade .

Example 1 Λ solution of 2.6 g of the benzyl ester of benzyl-30- (D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]carbamoyl-ethyl^-2-deoxy-2-propionylamino-«-D-glucopyranoside in 67 ml of 60% strength acetic acid is hydrogenated under normal pressure and at room temperature, using 0.6 g of 5% strength palladium-on-charcoal as the catalyst, until the reaction has ceased (hydrogenation time about 20 minutes). The catalyst is filtered off and rinsed with a little 60% strength acetic acid and the filtrate is evaporated to dryness under a waterpump vacuum. The residue is crystallised from ethanol/ether.

Mils gives benzyl-3-O-|D-2-fL-l~(D-l-carbamoyl-3-carboxy-propyl)carbamoyl-ethyl]-carbamoyl-ethyl^ -2~deoxy-2~propionylamino-«-Dglucopyranoside which has a melting point of 155-160° (decomposition) and an optical rotation.[α]^θ of « +105° -1° (dimethylformamide, c » 0.58).

The benzyl ester of benzyl-3-0~|D~l-[]>-l-(Dl~carbam©yl·3-carboxy-propyl)-carbamoyl-ethyl1-carbamoyl-ethylj-?.-deoxy-2propionylamino-cc-D-glucopyranoside which is used as the starting - 25 441®° material can be obtained as follows: 4.1 g of ben2yl-2-amino-4,6-0-bensylidene-5“0-(D-lcarboxyethyl)-2-deoxy-«-D-glucopyranoside are dissolved in 200 ml of methanol and 3 ml of triethylamine and 0.95 ml cf pro5 pionyl chloride is added dropwise at room temperature, whilst stirring. After stirring for 2 hours, the reaction mixture is evaporated, to dryness and the residue is dissolved in 100 ml of water. The pH of this solution is adjusted to 4 with icecold 2W hydrochloric acid and the product which has separated out is filtered off, rinsed with water and dried in vacuo over sodium hydroxide. The benzyl-4,6-0-benzylidene~3~0-(D-lcarboxyethyl)-2-deoxy-2-propionylamino-a-D-glucopyranoside which is thus obtained can be recrystallised from methanolj melting point 257°, [α]^θ = +132° -L° (dimethylformamide, c = 1.086). 1.5 g of N-ethyl-5-phenyl-isoxazolium 3'-sulphonate (Woodward's reagent K) are added to a solution of 3.1 g of the resulting compound in 150 ml of acetonitrile and 0.9 ml of triethylamine and the mixture is stirred at room temperature until everything has dissolved (about 60 minutes). 2.2 g of L20 aianyl-D-glutamic acid l-amide~7-ben?.yl ester-hydrochloride and 0.9 ml of triethylamine are now added and the mixture is stirred for a further 18 hours at room temperature. After distilling off the solvent under a waterpump vacuum, water is added to the residue and the insoluble matter is filtered off, washed thor25 oughly with water and dried. The resulting benzyl ester of ben2yl-4,6-0-benaylidene-3-0-^-l-[L-l-(D.-l~carbamoyl-3“0arboxypropyl)-carbamoyl-ethyl]-carbamoyl-ethy'4 ~2~deoxy-2-propionylamino- 26 44180 t'-D-glucopyranoside, which has an Rf value of 0.4(5 on silica gel thin layer platen in the system methylene chloride/methanol, 10/j, .is dissolved in 200 ml of 60% strength acetic acid and the solution is kept at S5~100°C for 1.5 hours. After cooling, the solvent is distilled off. The residue is taken up twice more in a little water and in each case is evaporated to dryness. The resulting benzyl ester of benzyl-3-0-^D-l-[L~ l-ill-l-carbxtmoyl-v-carboxy-propylj-carbainoyl-ethyl j-carbamoylethyl; “2-deoxy“.2-propionylamino-a-D~glucopyrano3idR is crystallised from methanol; melting point 189°, = +100° ^1° (dimethylformamide, c - 1.268).

Bxamp'le 2 A solution of 4 g of the benzyl ester of benzyl-2-acetamido-3-0-|rL-l-(D~l-carbamoyl-3-carboxy-propyl)-carbamoylethyl 3~carbamoyl-methy.l5 -2-deoxy-a-D-glucopyranoside in 80 ml of methanol is hydrogenated under normal pressure and at room temperature, using 0.4 g of 5% strength palladium-on-charcoal as the catalyst, until the reaction has ceased. The catalyst is filtered off and rinsed with a little methanol and the filtrate is evaporated to dryness under a waterpump vacuum. The residue is dissolved in 50 ml of distilled water and further hydrogenated under normal pressure and at room temperature, using 1 g of 5% strength palladium-on-charcoal as the catalyst, until the reaction has ceased. The catalyst is filtered off and rinsed with a little water and the filtrate is evaporated to dryness. Tho resulting 2-acetamido-3-0-^[L-l-(D-l-carbamoyl3-carboxyl-propyl)-carbamoyl-ethyl]-carbarcoyl-metbyl5-2-deoxy-D- 27 44190 glucose is dried over phosphorus pentoxide under a high vacuum? [k]q° = -10° ±1° (water, c = 0.930).

The starting material can be prepared as follows: ·3 g of N-ethyl-5-phenyl-isoxazolium 5'-sulphonate (Woodward's reagent K) are added to a solution of 9.5 g of benzyl2-acetamido-4,6-0-benzylidene-3-0-carboxymethyl-2-deoxy-a-Dglucopyranoside in 400 m3, of acetonitrile and 3 ml of triethylamine and the mixture is stirred at room temperature until a clear solution forms. 7.15 g of L-alanyl-D-glutamic acid ΙΙΟ · amide-T-benzyl ester-hydrochloride, 3 ml of triethylamine and 200 ml of acetonitrile are now added and the reaction mixture is stirred for a further 18 hours at room temperature. The benzyl ester of benzyl-2-acetamido-4,6-0-benzylidene-3-0-£[-L-l-D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethylJ~c&rbamoyl-methyl| -215 deoxy-a-D-glucopyranoside, which has crystallised out, is filtered off, washed with half-saturated sodium bicarbonate solution and water and dried; = +81° -1° (dimethylformamide, c = 0.816).

A solution of 8 g of this compound in 400 ml of 60Ji strength acetic acid is kept at 80°C for 1 hour. After cool20 ing, the solution is evaporated to dryness, a further twice 50 ml of water are added to the residue and in each case the mixture is evaporated to dryness. The resulting crystalline residue is stirred with a little water and the crystals are filtered off and dried. This gives the benzyl ester of benzyl-2-acetamido~3-025 ΐ[L-l-(D-l-carbamoyl-3-carboxy~propyl)-carbamoyl-ethyl]-aarhamoylmethylf-2-deoxy-K-D-glucopyranoside which has a melting point of 200-202°, ~ (dimethylformamide, c = 0.599). - 28 44130 Example 3 A solution of 0.9 g of the benzyl ester of benzyl-2ac etamido-3~0- [L-l- (D-l~carbamoyl--3-carboxy -propyl) -carbamoylethyl]-carbamoyl~methyl-2-deoxy-6~0~stearoyl-a-D~gluccpyranoside j.n 40 ml of methanol is hydrogenated under normal pressure and at room temperature, using 0.2 g of 5% strength palladium-oncharcoal as the catalyst, until 22.4 ml of hydrogen have been taken up. The catalyst is filtered off and rinsed with methanol and the filtrate is evaporated to dryness. This gives benzyl-2-acetaiDido-3“0-LL-l-(D-l-carbamoyl-3-carboxy-propyl)c.arbamoyl-ethylJ-oarbamoyl-methyl-2-deoxy-6~0“Stearoyl-«-Dglucopyranoside, [α]^θ = +33° -1° (chloroform, c = 1.046).

The starting material can be prepared as follows: A solution of 1.4 g of stearyl chloride in 7·5 ml of methylene chloride is added dropwise in the course of 1 hour to a solution of 2.8 g of the benzyl ester of benzyl-2-acetamido3-0-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]carbamoyl-methyl-2-deoxy-a-D-glucopyranoside in 30 ml of pyridine, whilst stirring and with the exclusion of moisture, and •the mixture is stirred at room temperature for 48 hours. The reaction mixture is poured into ice water and extracted with methylene chloride. The organic phase is washed with ice-cold 2N hydrochloric acid and water, dried over magnesium sulphate and evaporated to dryness. The residue is purified by column chromatography on silica gel by elution with ethyl acetate.

This gives the benzyl ester of benzyl-2-acetamido-3-0[L-X~(D-l~carbamoyl~3-carboxy-propyl)-carbamoyl-ethyl.] -carbamoyl- 29 441®° methyl-2-deoxy-6-0-stearoyl-a-D~glucopyranoside whic.h has an [«]^° of +30° ±1° (chloroform, c -- 1.203)· Example 4 Analogously to Example 2, benzyl-2-acetamido-4,6-Qbenzylidene--3“0-(D-l~carboxy-propyl)-2-deoxy-o;-D-glucopyr anoside is subjected to a condensation reaction with L-alanyl-D-glutamic acid l-amide-Y-benzyl ester-hydrochloride and ‘the protective groups are split off. This gives 2-acetamido~3-0~£D-l-[L~ 1-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoylpropyl^ -2-deoxy~D-glucose.

The starting material can be prepared as follows: g of sodium hydride are added to a solution of 60 g of benzyl-2-acetamido-4,6-0-benzylidene-2-deoxy-a-D-glucopyrano~ side in 600 ml of dimethylformamide and the mixture is stirred for 1.5 hours at 45°C in a nitrogen atmosphere. After · cooling to 0°, 75 ml of ethyl D,L-a-bromobutyrate are added.

The reaction mixture is stirred for 1 hour at room temperature and for 1 hour at 50°C and neutralised, with acetic acid and the solvent is evaporated under a waterpump vacuum. The residue is partitioned between methylene chloride and water, the organic phase is washed once more with water and dried over magnesium sulphate and the solvent is evaporated. The residue, which is dried under a high vacuum, is purified by column chromatography on silica gel. Elution with methylene chloride/ethyl acetate (85/15) gives the ethyl ester of benzyl-2-acetamido-4,6-0benzylidene-3-0-(D-l-carboxy-propyl)-2-deoxy-a-D-glucopyranoside which has an Γα]^° of +113° ±1° (chloroform, c = 0.5), a melting point of 15zt°C (from methylene chlori.de/ether) and an Rf value of 0.21 and tho ethyl ester of benzyl-2-acetamido-4,6-0-b®nzyli~ done-3-0-(L-l-carboxy~propyl)-2-deoxy~a-D-glucopyranoside which has an Γα'Ιρθ of +42° il° (chloroform, c = 0«511), a melting . point of 240°C (from ethyl acetate) and an Rf value of 0.04. 100 ml of IN sodium hydroxide solution are added to a solution of 38.1 g of the ethyl ester of benzyl-2-acetamido-4,6“ 0-benzylidene-3-0(D-l-caiboxy-propyl)-2--deoxy~K~D-glueopyr,anoside in 300 ml of methanol and the mixture is kept at S0J for 1 hour. Tlie solution is then cooled, concentrated to about 150 ml and diluted with 400 ml of ice water and 100 ml of a IN icecold hydrochloric acid are added. The product which has crystallised out is filtered off, washed with water and dried.

This gives benzyl-2-acetamido-4,6-0-benzyIidene~3-0-(D~l-carboxy~ propyl)~2~deoxy~K-D-glucopyranoside which has a melting point of 210-213° and an [κ)^θ of +110° -1° (dimethylformamide, c - 0.554-).

Benzyl-2-acetamido-A,6-0-benzylidene-3-0-(L-l-carboxypropyl)-2-deoxy-«-D-glucopyranoside which has a melting point of 2O5°C and an [<*3ρθ of +71° -1° (dimethylformamide, c .-.: 0-589) is obtained in an analogous manner.

Example 5 3.77 g of 2-phenyl-4,5“[3-0-(D-l-carboxyethyl)~5,6“0~ isopropylidene-D-glucofurano]-4^-oxazoline in 60 ml of acetonitrile and 15 ml of dimethylformamide are stirred together with 1.4 ml of triethylamine and 2.55 g of N-ethyl-5-phenyl-isoxazolium 3'-sulphonate for 1.5 hours at 0°, whereupon virtually everything goes into solution. 3.44 g of L-alanyl-D-glutamic acid 1- amide-'f-benzyl ester-hydrochloride and a further 1.4 Ell of triethylamine are then, added and the mixture is stirred for 24 hours at room temperature. It is then evaporated under an oil vacuum to give a syrup and the latter is chromatographed on . silica gel using a mixture of chloroform and acetone (8/2).

A colourless, firm syrup which crystallises on grinding with ether is obtained. Melting point 56-99°! Εβ]ρ° - +13° (ih chloroform).

The crystalline benzyl ester is hydrogenated, using 5% · strength palladium-on-charcoal, in dioxane at room temperature and under normal pressure and after evaporating in vacuo gives the parent acid as a syrup.

This is stirred in watei' with 10 ml of Eowex-50-H+ foils hours at room temperature. After filtering and freeze15 dryingp a colourless powder with a decomposition temperature of 140° is obtained. The 2-benzoylamino-3-0-|-D-l-[L-l-(D-lcarbamoyl“3-carboxy-propyl)-l-carbamoyl~ethyl3-carbamoyl-ethy4 · 2- deoxy-a.β-D-glucose which is obtained contains varying amount! of water of crystallisation, depending on the drying conditions and in the above case after drying at 60° and under 0.01 mm Hg for 15 hours contains 1/3 water.

Example 6 6.0 g of 2-phenyl-4,5~[3“0-(D-l-carboxypropyl)-5,6-0Λ isopropylidene-D-glucofurano]-A -oxazoline, 4.08 g of N-ethyl25 5-phenyl-isoxazolium 3'-sulphonate and 2.25 ml of triethylamine are stirred in 100 ml of acetonitrile and 25 ml of dimethylformamide for 1 hour at 0-5°C, whereupon the whole goes into - 32 44190 solution. 5«55 g of L-alanyl-D-glu’fcamic acid l-amide-γbenzyl ester-hydrochloride and a further 2·35 ml of triethylamine are then added and the- mixture is stirred at room temperature for 48 hours. It is evaporated under an oil vacuum and chromatogi-aphed. on silica gel in a mixture of chloroform and ethanol (19/1). 9 g of the colourless syrup which is thus obtained are hydrogenated in dioxane using 5% strength palladium-oncharcoal, the catalyst is filtered off and the filtrate is concentrated in vacuo and hydrolysed at room temperature in a mixture of 40 ml of tetrahydrofurane and 30 ml of water with 1.5 m'.l of trifluoroacetic acid. She water is then evaporated to dryness 4 times in vacuo and the residue is dissolved in water and lyophilised. The resulting 2-benzoylamino~3-0“^D-l-[L-l(D-l-carbamoyl~3~carboxy-propyl)-carbamoyl~ethyl]-carbamoylpropyl).-2-deoxy-a.β-D-glucose crystallises with 0.5 mol ef water, melting point 114-152°, [α]ρθ - +17° (ih methanol).

Example 7 3.63 g of 2-phenyl-4,5-[3-0-carboxymethyl~5,6-0-isopropylidene-D-glucofurano]-42-oxazoline, 3.43 g of L-alanyl-Dglutamic acid l-amide-T-benzyl ester-hydrochloride, 1.21 g of Nhydroxysuccinimide, 2.16 g of dicyclohexylcarbodiimide and 1.45 ml of triethylamine are dissolved in 40 ml of dimethylformamide and the mixture is stirred for 24 hours at room temperature.

It is evaporated under an oil vacuum, the residue is taken up in dichloroethane and water, the dicyclohexylurea which has precipitated out is filtered off and the organic phase is twice extracted by shaking with water and the aqueous phase is twice 44ΐθ° extracted by shaking with dichloroethane. After drying and evaporating the organic phases, a solid syrup is obtained and this is chromatographed on silica gel. in a mixture of chloroform/ethanol (9/1). The resulting peptide ester, which crystallises on grinding with ether, melts at 167-163°, [ce]p° = “5° (chloroform). 4-5 g of the said ester are hydrogenated in dioxane using 5% strength palladium-on-charcoal and the catalyst is filtered off and further extracted with'ethanol. The combined filtrates are evaporated and the residue is recrystailised from isopropyl alcohol. The resulting acid melts at 200-207°. 2.85 g of this acid, in a mixture of 30 ml of water and 15 ml of tetrahydrofurane, are stirred with 5 ml of I)owex~50~H+ for 15 hours at room temperature, the mixture is filtered through a hardened filter and the filtrate is evaporated to dryness in vacuo. On grinding with ether a colourless powder is obtained and this is 2-b€aizoyl-amino-3-0-|[L-l-(D-l-oarbamoyl-3-oarboxypropyl)--l-carbamoyl~ethyl-carbainoyl-methyl^ -2-deoxy-a. β-Dglucose which has a melting point of 175-177° (as the hydrate).

The starting material can be prepared as follows, using a modification of the method described in Acta Chem. Scand. 18, 185 (1964): 100 g of 2-phenyl-4,5~[5»6-0-isopropylidene-D-glucoο furano]-Δ -oxazoline are dissolved in 1 litre of acetonitrile, with the exclusion of moisture and carbon dioxide, and 15.2 g of a 55% strength NaH/mineral oil dispersion are added in portions, whilst stirring well, and the mixture is stirred for a - 34 44190 fur’'ή er one hour at room temperature. 42 ml of ethyl chloroacetate are then added, dropwise at 0° and after 1.5 hours a further 42 ml are added dropwise. After 1.5 hours, a further 11.4 g cf the Kali dispersion are added, the mixture is stirred for half an hour and a further 42 ml of the chloroacetate are added dropwise at 0°. After a further 2 hours, the mixture is allowed to warm to room 'temperature and is evaporated ir. vacuo - towards the end under an oil vacuum - to give a syrup. The latter is taken up in ether and extracted 3 times by shaking vzith water, the ether phase is dried over sodium sulphate and, after evaporating, 155 g oi a brown oil are obtained. This is dissolved in 150 ml of methanol and a solution of 30 g of potassium hydroxide in 150 ml of water is added, tlie mixture j {i twice extracted with ether and the ether phase .is washed once with water. The aqueous phases are freed from ether in vacuo and the pH is adjusted to 4 with IK hydrochloric acid, using a pH meter.

The crystalline 2-phenyl-4,5-[3-0-(earboxymethyl)-5,6p 0-isopropylidene-D-glucofurano]-4 -oxazoline which has precipitated out is filtered off, washed with water and dried. 107 g, that is 93% of theory, with a melting point of 186-188°, an cf -9° (CHCly c = 0.9) and an [a]^0 of -23° (CHCly c = 3) are obtained.

I Example 8 2.5 g of 2-benzamido-2-de0xy-3-0-[L-l-(D-l-carbamoyl~3carboxy-propyl)-carbamoylethyl]-carbamoylmethyl-D-glucopyranose (Example 8) are dissolved in a mixture of 17 ml of absolute - 35 “ 44ΐθ° pyridine and 4 ml of butyric anhydride. After 20 hours at room temperature and 2 hours at 50°j water is added and the raixture is concentrated in vacuo. The residue is dissolved in chloroform, the chloroform solution is extracted by shaking with IN hydrochloric acid and water and dried over sodium sulphate and the chloroform phase is evaporated. The oily residue is extracted several times with ether and a solid amorphous mass with a melting point of 110-120° is thus obtained; this is κ,β1,4,6-tri-buiyrcyl -2-benzamido-2-deoxy-3-0-[L-l“(D-l-carbamoyl-3~ carboxy-propyl)-carbamoyletbyl]-carbamoylmethyl-D-glucopyranose, R, - 0.J2 in CHC1,:CHXQR - 7:2 (silica gel thin layer, Merck). Example 9 2.3 g of 2~benzamido-2~deoxy-3“0~[L-l“(D-l~carbsmoyl3 carboxy-propyl)-carbamoylethyl]-carbamoylmethyl-D-glucopyranose are dissolved in 20 ml of pyridine and 5 ml of acetic anhydride. After 3 hours at room temperature, water is added and the mixture is concentrated in vacuo. It is purified over 70 g of silica gel Using CHCl^CH^OH - 3:1 and a colourless powder which has a melting point of 122-158° and an [a]^0 of +48° (CH^OII, c = 1.074) is obtained; this is a.p~l,4,6“triacetyl-2-benzamido-2-deoxy3-0-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoylethyl]carbamoylmethyl-D-glucopyranose, R^ 0.54, CHCl^CH^OH = 3:2 (silica gel thin layer, Merck).

Example 10 6·33 g of 2-phenyl-4,5“(3-0-carboxymethyl-5,6-0-isop propylidene-D-glucofurano)-A -oxazoline, 5.75 g of 2-ethoxy-Ncarbethoxy-l,2-dihydrcquinoline (EEDQ) and 9-3 ml of triethyl- 36 44190 amine are added to a solution of the trifluoroacetate of dibenzyl L-alanyl-D-glutamate (obtained from 8.3 g of dibenzyl N-tert.butoxycarbonyl-L-alanyl-D-glutamate and 5.1. ml of trifluoroacetic acid and 2.6 ml of dichloroethane by hydrolysing for 4 hours at 40°) in 70 ml of dichloroethane. The mixture is allowed to react at 40° for 15 hours and is diluted with chloroform and twice extracted by shaking with water and the aqueous phases are extracted once by shaking with chloroform. After drying ovex' sodium sulphate and evaporating the chloroform solution, 19.9 g of an oil are obtained and this is purified over 400 g of Merck sili.ca gel by elution with ether and then with chloroform:acetone, 17.‘3· This gives pure 2-phenyl-4,5“ [3-0-( 1-1,-1-D,3-dibenzyloxy-carbonyl-propylJ -carbamoylcthyl)carbarcoylraethyl-5>6-Q~i3opropylidene-D-gluco£ux,anoJ~A4'~oxa2oline which has a melting point of 113-116° and an [a]^° of -47° (CHCly c - 1.54). g of the above compound are hydrogenated in a mixture of 80 ml of tetrahydrofurane and 20 ml of water, using 1.8 g of 5% strength Pd/C, until the reaction has ceased, the catalyst is filtered off, the filtrate is evaporated in vacuo and the residue is ground with ether. This gives 4.9 g of the dicarboxylic acid as a colourless powder. 4.4 g of the above dicarboxylic acid are stirred with . ml of Dowex-50 W x 4 ion exchange!' in a mixture of 45 ml of tetrahydrofurane and 20 ml of water for 20 hours at 40°. After filtering and clarifying with charcoal (Darco G 60), the solution is freeze-dxied and colourless, amorphous 2-benzamido-2-dooxy-3.. 57 .. 44ΐ«θ O-[l-L-(l-D,3-dicarboxy~propyl)-carbamoyl-ethyl]-carbamoylmethylD-glucopyranose which has an optical rotation of ί-α3ρθ - +25° (HgO, c = 0.997), is obtained.

Example 11 . Analogously to Example 10, 5-7 g of 2-phenyl-4,5.13-0carboxymethyl-5,6-0-isopropylidene~D-glucofurano]-Δ -oxazoline are subjected to a condensation reaction with 4.9 g of L-serylD-glutamic acid ά-amide-T-tert.-butyl ester-hydrochloride with the addition of 2.3 ml of triethylamine and 5.2 g of 2-ethoxy-Ti10 · carboethoxy-1,2-dihydroquinoline in 4-5 ml of dichloroethane.

After 18 hours at 40° a crystalline product has formed. A further 5θ nil of dichloroethane are added and the mixture is cooled in ice and filtered and the crystals are washed with cold dichloroethane. The analytically pure, colourless crystals of 2-phenyl-4,5-[3-0-(1-L-£l-D-carbsmoyl-3-tert.-butyloxycarbonyl-propylj-carbamoyl-2-hydroxyethyl) -carharaoylinethyl-5,6p 0-isopropylidene-D-glucofu.rano]~A -oxazoline have a melting point of 187-183° and a [ct]^0 of +7° (CI^OH, c = 1.125). g of this compound are hydrolysed with a mixture of ml of methylene chloride and 5 ml of trifluoroacetic acid for . hours at room temperature. The mixture is evaporated under an oil vacuum, the residue is ground with ether and 2-benzamido2-deoxy-3-0-[l-L-(l-D-carbamoyl-5-carbcxypropyl)-carhamoyl-2hydroxyethyl]-carbamoylmethyl-D~glucopyranose is obtained as a beige powder with a melting point of 100-115°, = +23° (HgO, c = 0.886), which crystallises with 2 mols of water and 1 moi of trifluoroacetic acid. R.f = 0.28 CHCl^CH-OH = 1:1 - 38 44180 (silica gel thin layer, Merck).

Example 12 Analogously to Example 10, 5.25 g of 2-phenyl-4,5“-[3p O-carboxymethyl-5,6-0-isopropyli dene-D-g'J.ucoiP.ra?To]-Λ -oxazoline and the trifluoroacetic acid salt of L-alanyl-D-glutsmic acid nr-n-px'opylciinide-y-bfcnzyl ester - obtained from 6.2 g of N-tert.butoxy-carbonyl-L-alanyl-D-glutamic acid «-n-propylamide-Y-benzyl ester and 4.2 ml of trifluoroacetic acid in 2-5 ml of dichloroethane after 6 hours at 40° - in 60 ml of dichloroethane are subjected to a condensation reaction, with the addition of 7.75 ml of triethylamine and 4.8 g of 2-ethoxy-N-carboethox5f-l,2~ dihydroquinolj.ne. After 20 hours at 40°, the mixture is diltited with 50 ml of chloroform and twice extracted by shaking vzith water and the water is twice extracted by shaking with chloroform. After drying and concentrating the chloroform phase, g of un oil are obtained and this is purified over 200 g of Merck silica gel by elution with ether and then with chloroform: acetone = 7:3· 6.4 g of a colourless amorphous substance with an of Ο.35, CHClacetone = 7:3 (silica gel thin layer, Merck) are obtained.

This substance is hydrogenated in 80 ml of tetrahydrofurane and 20 ml of water, using 1.8 g of 5% strength Pd/C, until the reaction has ceased, the catalyst is filtered off and the filtrate is concentrated. The acid has an Rf of Ο.58, CHC1^:CH^OH - 3sl (silica gel thin layer, Merck). It is then stirred with 10 ml of Dowex 50-W x 4 ion exchanger, 50 ml of tetrahydrofurane and 25 ml of water for 15 hours at room temperature and for 12 hours at 40°. After filtering, clarifying the filtrate with charcoal (Dareo-G-δθ), filtering again and freeze-drying the filtrate, colourless, amorphous 2-henzamido-2-deoxy-3-0- [l-L- (l-D-N-n-pr opyL-carbamoyl-3-carboxy-propyl) 5 . carbamoylethyl]-carbamoyl-methyl-D-glucopyranose with a melting point of 65-140° and an of +28° (water, c = 1.03) and an R^ of 0.48, CHClyCH^OH ~ 1:1 (silica gel thin layer, Merck) is obtained.

Example 13 7·3 g of 2-phenyl-4,5-[3-0-carboxymethyl-5,6-0-isopropylp idene-D~glucofurano]~4 -oxazoline, 6.5 g of a-amino-isobutyroylD-glutamic acid α-amide-T-tert.-butyl ester-hydrochloride and 2.9 g of isobutyl chloroformate 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 to this solution at -150 to -10° in the course of 30 minutes.

Tlie mixture is then allowed to warm to room temperature and is stirred for a further 15 hours at room temperature. It is diluted with 50 ml of dichloroethane and extracted by shaking with water, twice by shaking with 0.5 N NaOK and three times by shaking with water and the aqueous phases are extracted by shaking twice with dichloroethane, the organic phases are dried and, after evaporating, 16.6 g of an oil are obtained. This is purified over Merck silica gel by elution with CHCl^CgH^OH - 19:1. 9·7 g of colourless, amorphous 2-phenyl-4,5methyl-l-(l-D-carbamoyl-3-tert.-butoxycarbonyl-propyl)-carbamoylethyl ]-5,6-0-isopropyiidene-3-glucofuranoj-Δ^-oxazoline which has - 40 44190 an optical rotation: - +6° (CHCl-, c ~ 1.027), a melting point of 75-89° and an Rg of 0.35, CHClyCgH^OH - 9:1 (silica gel thin layer, Merck) are obtained. 8.3 g of the above compound are allowed to stand in a mixture of 20 ml of trifluoroacetic acid, 60 ml of methylene chloride and 2 ml of water for 15 hours at room temperature.

The mixture is then evaporated in vacuo and the residue is ground with ether. The resulting pink-coloured powder is dissolved in 200 ml of water and the solution is clarified with 0.5 g of Darco-G-60 charcoal. After filtering and evaporating the filtrate, colourless, amorphous 2-benzamide-2-deoxy-3-O-Llmethyl-l~(D-l-carbamoyl-3“Carboxy-propyl)Carbamoylethyl]carbamoylmethyl-D-glucopyranose with a melting point ef 110-120°, an [g]q° of <51° (HgO, c - 0.88) and an Rf of 0.52, acetone: ethanol = 1:1 (silica gel thin layer, Merck), which crystallises with 0.6 mol of trifluoroacetic acid and 1.7 mols of water, is obtained.

Analogously to Example 10, 2-benzamido-2-dooxy-3-0carboxymethyl-^-ethyl-D-glucopyranoside and the trifluoroacetic acid salt of L-alanyl-D-glutamic acid a-amide-7-tert.-butyl ester with 2-ethoxy-N-carboethoxy-l,2-dihydroquinoline give the corresponding glycopeptide which has an [κ]ρof -23 (CH7OH, c = 1.107) and an Kf of 0.47 (CHgCl^CgH^OH - 3:2) and, after hydrogenation in tetrahydrofurane/wator using 5% strength Pd/C, the glyao pgjtide gives 2~benzamido~2-deoxy-3-0-[L-l-(D-l~carbamoyl~3“ carboxy-propyl)-carbamoylethylJ-earbamoylmethyl-p-ethyl-D- 41 441-90 glueopyranoside which has a melting point of 215-217°, an [a]^0 of -22° (CH30H, c = 0.97) and an Rf of 0.36 in CHCljZGHjOH = 1:1 (silica gel thin layer, Merck). 2-Benzamido-2-deoxy--3“0-car'boxymethyl-p-ethyl-'D-gluco·-pyranoside which is used as the starting material is obtained as follows: 2-Phenyl-4,5-[3-0-carboxymethyl-5,6-0-isopropylidenep D-glucofurano]-A -oxazoline is dissolved in 0.1 N HCl/CgH^OK and the solution is left to stand for 6 hours at room temperature. It is neutralised with sodium ethylate in ethanol and evaporated to dryness and the residue is taken up in acetone.

The solution is filtered through a layer of Merck silica gel, the eluate is evaporated to dryness and the residue is extracted twice with ether at room temperature. After recrystallisation from ethyl acetate, 2~benzamido~2-deoxy-3-0-carboethoxymethyl-pethyl-D-glucopyranoside which has a melting point of 185-188° and an [a]^0 = -35° (CH^OH, c = 1.121) is obtained. 9-4 g of this ester are saponified with a solution of 1.7 g of potassium hydroxide in 250 ml of ethanol and 25 ml of water foi’ 2 hours at room temperature. The pH value is then adjusted to 3.5 with 3. N hydrochloric acid and the mixture is concentrated in vacuo.

The residue is first ground with ether and is then ground with 3 times 20 ml of ice water and the mixture is filtered. Crystals of 2-benzamido-2-deoxy~3-0-carboxymethyl-p-ethyl~Dglucopyranoside with a melting point of 205-210° and an of -40° (CH^OH, c = 1.04) are thus obtained. - 42 44190 Example 15 2~Benzemida-2-deoxy~5~D-i (D-I-carbamoyl-jS-carboxy·. propyl)-carbamoylmethyl3-carbamoylmexhyl-D-glucose is obtained from 3 g of 2-phenyl-4,5-[5-0~j(D~l~carbamoyl-3-carboxy-propyl)5 carbamoyimethylj-carbamoylmethyl-5>6-0~isopropylidene-D~glucop furano]-Λ -oxazoline by hydrolysis with 1.5 ml of trifluoroacetic acid in a mixture of 15 ml of dimethoxyethane and 15 ml of vzater at 40° for 3 hours. The reaction mixture is concentrated to dryness in vacuo and the residue is again extracted . with ether. The residual powder is dissolved in vzater and the solution is treated with Darco-G-60 charcoal and filtered and the filtrate is freeze-dried. This gives a colourless, amorphous substance with a melting point of ΙΙ5-Ί550 and an [α]β° of +34° (water, c = 0.81), Rf = 0.28 CHCKtCHjOI-I = 1:1 (silica gel thin layer, Merck).

The starting material for this reaction is obtained as follows: 8.0 g of N-terfc.-butoxycarbonyl-glycyl-D-glutamic acid a-amide-y-benzyl ester are dissolved in a mixture of 6.3 ml of trifluoroacetic acid and 7 ml of dichloroethane and the solution is allowed to react for 2 days at room temperature and for 3 hours at 45°. 12.1 ml of triethylamine, 7.0 g of 2-ethoxy-Ncarboethoxy-l,2-dihydroquinoline (EEDQ) and 8.1 g of 2-phenylp 4,5~[3-0-carboxymethyl-5,6-0-isopropylidene-D-glucofuranol)-425 oxazoline and 20 ml of dimethylformamide are added to the solution, whilst cooling. After 20 hours at 40°, the mixture is concentrated under an oil vacuum and the residue is partitioned 44ΐθ° between methylene chloride and water. After drying and concentrating the methylene chloride phases, a solid residue is obtained and this is extracted twice with ether and recrystallised from toluene. Yield 8.25 gi melting point 157°, [a]p° , = +10° (CHClj, c = 1.48), Rf ~ 0-35 (CHCly.ethanol = 9:1) (silica gel thin layer, Merck).

The benzyl ester vzhich is thus obtained is hydrogenated in 100 ml of tetrahydrofurane and 25 ml of water, using 1 g of 5% strength Pd/C, until the reaction has ceased. After fillo tering off the catalyst and evaporating the filtrate, the substance is chromatographed on 250 g of Merck silica gel in CHCljiCH^OH = 4:1. 5.8 g of colourless, amorphous 2-phenyl4,5~[3-O-!(D-l-carbamoyl-3-carboxy-propyl)-carbamoylmethylj carbamoylmethyl-5,6-0-isopropylidene-D-glucofurano]-Δ -oxazoline which has an R^. of 0.43, CHCl^tCH^OH = 3:2 (silica gel thin layer, Merck) are obtained.

Example 16 Analogously to Example 10, 9-5 g of 2-phenyl-4,5--[3~ 0-carboxymethyl~5,6-0-isopropylidene-D-glucofuranoΙ-Δ^-oxazoline are subjected to a condensation reaction with 6-25 g of Lalanyl-D-glutamic acid α,γ-diamide-hydrochloride with the addition of 3.4 ml. of triethylamine and 7.95 g of 2-ethoxy-Ncarboethoxy-1,2-dihydroquinoline (EEDQ) in a mixture of 50 ml of dichloroethane and 150 ml of dimethylformamide. The mix25 ture is allowed to react, whilst stirring, for 2 days at room temperature and for 4 hours at 40°. It is concentrated under an oil vacuum and the residue is extracted, first -twice with - 44 44190 ether and then twice with ice water. After drying, the product can be recrystallised from dichloroethane. This gives colourless crystals which have a melting point of 170-3.64°; ta'jg0 - +3° (DMSO, c = 1.43), Rf = 0.64 CHCljSCHjOH = 3:1 (silica gel thin layer, Merck). 6.1 g of this compound are hydrolysed with 13·5 ml if Dowex 50 ion exchanger in a mixture of 60 ml of dimethoxyethane ar.d 60 ml of water for 15 hours at room temperature. After filtering and concentrating the filtrate, the residue is taken up in water, the mixture is clarified with Darco-G-60 charcoal and filtered and the filtrate is freeze-dried. This gives colourless, amorphous 2-benzamido-2-deoxy-3-0-[L-l-(D-l,3dicarbamoyl-propyl)-carbamoylethyl]-carbamoylffiethyl-D-gluco~ pyranose which has a melting point of 82-143°; [a]qC = -)-24° (II?0, c = 0.98), Rf - 0.45 CIKHyCHjOH = 1:1 (silica gel thin layer, Merck). The substance crystallises vzith 1.23 mols of water of crystallisation.

Example 17 Analogously to Example 16, ethyl-2-benzamido-2--deoxy3-0-carboxymethyl-p--D-glucopyranoside and L-alanyl-D-glutamic acid α,γ-bis-methylamide-hydrochloride give p-ethyl-2-benzamido2-decxy-3-0-[L-l-(D--l,3-bis-W-methyl-carbamoyl-propyl)-carbamoylethyl j-carbamoylmethyl-D-glucopyranoside vzhich has a melting point of 233°, an [α]^θ of -20° (CH-jOH, c = 0.937) and an Rf of Ο.38 in CHCljSethanol, 7:3 (.silica gel, thin layer plates, Merck). - 45 Example 18 Analogously to Example 16> 2-benzamido-2-deoxy-3-0carboxymethyl-D-glucopyranose gives 2-benzaraido-2-deoxy-3-0[L-1-(D-1,3-bis-N-rnethylcarbamoyl-propyl)-carbamoylsthylj5 carbamoylmethyl-D-glucopyranose which has a melting point of 125-13?-°, and [a]^° of and an Rf of 0.26 in CHCOy ethanol = 7:3 (silica gel thin layer plates, Merck).

Example 19 Analogously to Example 16, 2-benzamido-2-deoxy-3-010 · carboxymethyl-D-glucopyranose and the hydrochloride of dimethyl. L-alanyl-D-glutamate give 2-benzamido~2~deoxy-3-0-[L-l-(D-l,3biS“methoxycarbonyl-propyl)-carbamoyl~ethyl]“Carbamoylmethyl-Dglucopyranose in the form of the hydrate with a melting point of 60-90°, an [a]p° of +25° (CH-jOH, c = 1.017) and an Rf value of 0.23 in CHCl^:ethanol = 9:1 (silica gel thin layer plates, Merck).

Example 20 Analogously to Example 16, ethyl-2~benzamido-2~d.eoxy-3~0carboxyraethyl-g-D-glucopyranoside gives ethyl-2-benzamido-2~ deoxy~3~0~[L-l-(D-l,3-bis-niethoxy-carbonyl~propyl)-carbamoylethyl] carbamoylmethyl-p-D-glucopyranoside which has a melting point of 127-135°, an [a]^0 of -17° (CH^OH, c = 1.024) and an Rf of 0.26 in ethyl acetate:acetone = 2:1 (silica gel thin layer plates, Merck).

Example 21 .2-Benzamido~2~deoxy'73“0~[L-l-('D-l-N-behzyl~carbsmoyl“3 carboxy-propyl)-carbamoylethyl]*carbamoylmethyl-D-glucopyranose is obtained analogously to Example 12. - 46 44180 In this Ci-se, the l-carboxyl group cf rhe glut -.Pc ,ιοΐά i:: liberated from a tert.-bruyl estcx· by saponificatiw- with Dovex-bO-if in tetrahydrofurene/weter.

Example 2? 2-Benzamido-2-deoxy~3-0-[L-l-(D-l-R-carbaffioylmcihylcarbamoyl-3-carboxypropyl)'-earbamoylethyl]-carbrxmoylmethyl-Dglucopyranose is obtained analogously to Example 12. The starting material for the peptide part is K-tert.-butoxycarbonylL-alanyl-D-glutamic acid γ-benzyl ester-a-glycine-amide.

Example 23 Analogously to Example 12, the hydrochloride of L-kijaino-valsroyl-D-glutamic acid a-amide~7-tert.-butyl ester gives 2~benzan)ido-2~deoxy-3--0-' fL-l- (D-l-oarbamoyl-3-carboxy-propyl) carbamoylbutyl ‘J-carbamoylmethyl-D-glucopvranose.

Example ,-.4 2-Benzamido-2~deoxy~3~0-[L-l-(D-l-carbamoyl-3--carboxypropylJ-carbamoyl-propylj-carbamoylmethyl-D-glucopyranose is obtained analogously to Example 12.

Example ?3 2-Benzamido-2-deoxy-3-Ci-[L-l-(D-l“carbamoyl~3-c::irboxypropyl) -carbainoyl’2-inethylpropylj -l-carbamoylmethyl-I>.gTncciF5rraxosc is obtained analogously to Example 12.

Example 26 A solution of 10-7 g of the benzyl ester of benzyl-2acetaniido-4,6-0-benzylidene~3“0-[(D“l“Carbainoyl“3~carboxypropyl)-carbamoylmethyl-carbamoylmethyl]-2~deoxy-a-D-glucopyranoside in 200 ml cf glacial acetic acid and 100 ml of water - 47 441®° is hydrogenated under normal pressure and at room temperature for 57 hours using 2 g of 5% strength palladium-on-charcoal.

The catalyst is filtered off and rinsed with water and the filtrate is evaporated. The residue is taken up in water and the mixture is filtered through 100 ml of Amberlite IR 120 (H+ form) ion exchanger and the filtrate is freeze-dried. 2-Acetamido3-0-[(D-l-carbamoyI-3-oarboxy-propyl)-carbamoylmethyl-carbamoylmethyl]-2-deoxy-D-glucose is crystallised from methanol/ethyl acetate and dried under a high vacuum. The product, which contains l/4 mol of ethyl acetate, displays an optical rotation [aj§° = +27° (water, c = 0.944).

The starting material used can be prepared as follows: g of the benzyl ester of N-t-butoxycarbonyl-glycyl-P-isoglutamine are dissolved, at room temperature and vzith the exclusion of moisture, in a mixture of 18 ml of 1,2-dichloroethane and 8.4 ml of trifluoroacetic acid and the solution is left to stand for 16 hours. The reaction mixture is now diluted with 200 ml of tetrahydrofurane and, with external cooling, neutralised with triethylamine and a solution of 8.3 g of benzyl-2-acetamido20 4,6-0-benzylidene-3-0-carboxy-methyl-2-deoxy-a-Dglucopyranoside in 100 ml of tetrahydrofurane and 2.52 ml of triethylamine is added. After adding 5·Ο5 g of 2-ethoxy-N-ethoxycarbonyl-l,2dihydroquinoline (EEDQ), the mixture is stirred for 24 hours at room temperature. The product which has crystallised out. is filtered off, washed with tetrahydrofurane and ether and dried. The resulting benzyl ester of benzyl-2-acetamido-4,6-0-benzylidene-3-0-[(D-l-carbamoyl-3-carboxypropyl)-carbamoylmethyl- 48 44190 carbamoylmethyl ]-2-deoxy-...-D-glucepyranosi do has an opt leal rotation ~ +66° -1° (Ν,Ν-dimethylformamide, c -- 1.308).

Example 27 Λ solution of 4.5 g of the benzyl ester of methyl-2acetamido-3-0- [ [L-l- (D-l-carbanioyl-3-earboxypropyl) -carbamoylethyl ]-carbamoylmethylj -2-deoxy-a-D-glucopyranoside in 125 ml of 50% strength aqueous methanol is hydrogenated under normal pressure and at room temperature, using 1.0 g of 5% strength palladium-on-cliarcoal, until 178 ml of hydrogen has been taken up. The catalyst is filtei-ed off and. the filtrate is evaporated under a waterpump vacuum. The residua is dissolved in 50 ml of distilled water and freeze-dried. This gives methyl?.~&ce'tamido--3-0~i[L-l~(D~l~carbamoyl-3-carboxypropyl) -carbamoylethyl ]-carbamoylmethylj -2-deoxy-a-D-glucopyranoside which has an L«]p° of +49° ±1° (water, c = 0.939)· The starting material can be prepared as follows: 8.1 g of the benzyl ester of N-t-butoxycarbonyl--1.alanyl-D-isoglutamine are dissolved, at room temperature and • with the exclusion of moisture, in a mixture of 8.1 ml of 1,2dichloroethane and 8.1 ml of trifluoroacetic acid and the solution is left to stand for 16 hours. The reaction mixture is diluted with 200 ml of tetrahydrofurane and, with external cooling, neutralised with triethylamine and a solution of 7.62 g of met.hyl-2~aeetamido-4,6-0-benzylidene-3-0-carboxymethyl-2~ deoxy-a-D-glucopyranoside and 2.77 ml of triethylamine in 100 ml of tetrahydrofurane is added. After adding 5-0 g of 2-ethoxyN-ethoxycarbonyl-l,2-dihydroquinoline (EEDQ), the reaction _ 49 mixture is warmed to 40°, stirred at this temperature for 30 hours and left to stand for a further 24 hours at room temperature. The product vhich has precipitated out, that is to say the benzyl ester of methyl-2-acetamido~4,6-0-benzylidene-3-0-^[L5 l-(D--l-carbamoyl~3~carboxypropyl)-carbamoyl-ethyl]-carbamoylmethyl]-2-deoxy-oc-D-glucopyranoside, is filtered off, washed with tetrahydrofurane and ether and dried, [cc]^0 = +58° “1° (Ν,Ν-dimethylformamide, c = 1.125).

A solution of 10.5 g of the benzyl estei- of methyl-210 ac etamido-4,6-O-benzylidene-3-O-^[L-l-(D-l-carbamcyl-3-carboxypropyl)-carbamoyl-ethyl]-carbamoyl-methyl] -2-deoxy-a-D-glucopyranoside in 320 ml of glacial acetic acid is diluted with 200 ml of water, whilst stirring, and the whole is stirred at 50-55°0 for 2 hours. After cooling, the solution is evaporated to dryness, a further four times 100 ml of water are added to the residue and, each time, the mixture is evaporated to dryness.

This gives the benzyl ester of methyl-2-acetamid.O-3~0~][L-l(D-l-carbamoyl-3-cax'boxy-prqpyl)-carbamoyl-ethyl]-carbamoylmethyl]-2-deoxy-a-D-glucopyranoside which has an [σ,]^0 of +64° ^1° (Ν,Ν-dimethylformamide, c = 1.268).

Example 28 A solution of 3«0 g of the benzyl ester of methyl-2acetamido-3-0-^[L-l-(D-i-carbamoyl-3~carboxypropyl)-carbamoylethyl] -carbamoyl-methyl] -2-deoxy-6-0-stearoyl-a-D-glucopyranoside in 100 ml of ethanol and 100 ml of tetrahydrofurane is hydrogenated at room temperature and under normal pressure using 0.6 g of 5% strength palladium-on-charcoal, the catalyst is filtered - 50 44190 off ono the filtrate.· is evaporated to drynesj? under a waterpump vacuum. The crystalline residue is methyl-2-acetamido~ 3--0- J [L-l-(D-l-carbamoyl~3~earboxypropyl) -carbamoyl-ethyl ] carbamoyl-methyl} -2.-deo^-6-0~stearoyl-a-D-glucopyrano3ide which has an [α]^° of +50° ^1° (N,N-dimethylformamide, c. --- 0.921).

Tne starting material can be prepared as follows: A solution of 2.12 g of stearyl chloride in 20 ml of 1, 2- dichloroethane is added dropwise, at 0-5°, in the course of hours to a solution of 3· 98 g of the benzyl ester of methyl-2ac etamido-3-0-}[L-l~(D-l-carbamoyl-3-earboxy-propyl)-carbamoylethylj-carbamoyl-methylj-2-deoxy-a-D-glueopyranoside in 40 ml of absolute pyridine, whilst stirring and with the exclusion of iuoisiurej «nd the mixture is left to stand for 18 hours at room temperature. The reaction mixture is diluted with chloroform, washed with water, ice-cold 2 N hydrochloric acid and w&te-: dried over magnesium sulphate and evaporated to dryness. The product, that is to say the benzyl ester of methyl-2-acetamido3- 0-} (L-l- (D-l-carbamoyl-3-carboxypropyl) -carbamoyl-ethyl ‘J carbamoyl-methylj-2-deoxy-6-0-stearoyl-a-D-glucopyranoside, is crystallised from ethanol/ether; [a]^0 = +22° -1° (chloroform, c = 1.030).

Example 29 A solution of 6.8 g of benzyl-2-acetamido-3-0-}'[L~l(Π-1,3“dicarbamoylpropyl)~carbamoylethylJ-carbamoylmethylJ -2deoxy-a-D-glucopyranoside in 200 ml of 50% strength aqueous methanol is hydrogenated under normal pressure at room temperature for 60 hours using 5% strength palladium-on-chai’ccal. ~ 51 4419° The catalyst is filtered off and the filtrate is evaporated.

The residue is taken up in 50 ml of v/ater and lyophilised.

This gives 2-acetamido-3-0-^[L-l-(D-l,3-dicarbamoylpropyl)carbamoylethyl]-carbamoylmethyij-2-deoxy-D-glucose, which contains 1.24 mols of v/ater, in the form of a white powder with an [ccJpO of +7° —1° (water, c = 0.514).

The starting material can be prepared as follows: -0 g of L-alanyl-D-glutamic acid diamide-hydrochloride and 5·! g of 2-ethoxy-N-ethoxycarbonyl-l,2-dihydroquinoline (EEDQ) are added to a solution of 9.1 g of benzyl-2-acetamido4,6-0-benzylidene-3-0-carboxymethyl“2-deoxy-cc-D-glucopyranoside in 100 ml of Ν,Ν-dimethylformamide and 2-77 ml of triethylamine and the mixture is left to stand for 48 hours at room temperature. After distilling off the solvent, the residue is extracted thoroughly with ether and v/ater and the product is dried and recrystallised from chloroform/methanol: [o:]^0 +83° ±1° (Ν,Ν-dimethylformamide, c = 0.531).

A solution of 4 g of benzyl-2-acetamido-4,6-0~benzylidene3-0- [ [L-l- (D-l, 3-dicarbamoylpropyl) -carbamoylethyl J -carbamoylmethyl}-2-deoxy-a-D-glucopyranoside in 120 ml of glacial acetic acid is diluted with 80 ml of water and stirred for 3 hours at 60°. The reaction· mixture is then cooled and evaporated and a further 3 times 100 ml of water are added to the residue and the water is distilled off each time. The resulting benzyl2-acetamido-3-O- £ [L-l- (D-l,3-dicarb3moylpxpyl)-ca'bamoylethyl ]carbamoyl-methyl}-2-deoxy-a-D-glucopyranoside is recrystallised from methanol; melting point 223-225°. - 52 44190 A solution of 5.7 g of the benzyl ester of benzyl-2acetamido-3~0~/D-l-E(D-l'-carbaraoyl--3-carboxypropyl)-carbamoylmethylJ-carbamoylpropyl]~2-deoy.y~a-D-glucopyranoside in 100 ml of glacial acetic acid is hydrogenated in the presence of 5% strength palladium-on-charcoal under normal pressure and at room temperature. Ths catalyst is filtered off and. the filtrate is evaporated. The residual 2-acetaraido-3-0-^D-I-[(D-2-carbamcyl 3-carboxypropyl)-carbamoylmethyl3~carbamcylpropyl/ -2-deoxy-D10 glucose is taken up in 50 ml of water and freeze-dried; Μρθ = +46° ±1° (water, c - Ο.63Ο).

The starting material used can be prepared as follows: .1 g of the benzyl ester of N-t-butoxycarbonyl-glycyl-D-isoglutamine are dissolved, at room temperature and with the exclu15 sion of moisture, in a mixture of 5.1 ml of 1,2-dichloroethane and 5.1 ml of trifluoroacetic acid and the solution is left to stand for 16 hours. This solution is diluted with 100 ml of tetrahydrofuraue and, with external cooling, neutralised with triethylamine and a solution of 6.3 g of benzyl-2-acetamido-4,620 0~benzylidene~3“0--(D~l~carboxypropyl)-2~deoxy~tt-D-glucopyre.noside and 1.8 ml of triethylamine in 100 ml of tetrahydrofurane and 3.2 g of 2-ethoxy-N-ethoxycarbonyl-l,2-diliydroquinolins (EEDQ) is added. After Zl hours, the mixture is evaporated to dryness and the residue is partitioned between ethyl acetate and water. The organic phase is further washed with ice-cold 2 N hydrochloric acid, water, a saturated solution of sodium biear1 bonate and water and evaporated. The residual benzyl ester - 53 4419° of benzyl-2-acetaraido-4,6-0-benz\rlidene-3~0-^D-l-L(D-l-carbamoyl3-carboxypropyl)-carbamoylmethylJ-carbamoylpropylj-2-deoxy-a-Dglucopyranoside is crystallised from tetrahydrofurane/ether; [α]β° = +76° ±1° (Ν,Ν-dimethylformamide, c = 0.457).

. - Mild acid hydrolysis of this product in 60% strength aqueous acetic acid leads to the benzyl ester of benzyl-2acetamido-30-|D-l-[(D-l-carbamoyl-3~carboxypropyl)-carbamoylmethyl]-carbamoylpropylj-2-deoxy-a-D-glucopyranoside, which crystallises from methanol/efcher; melting point 180-185°, [α]^° = +87° (methanol, o = 1.035). g of the benzyl ester of N-t-butoxycarbonyl-L-a'lanylD-isoglutamine are dissolved, at room temperature and with the exclusion of moisture, in a mixture of 4 ml of trifluoroacetic acid and 4 ml of 1,2-dichloroethane and the solution is left to stand for 16 hours. This reaction mixture is now diluted with 30 ml of 1,2-dichloroethane and, with external cooling, neutralised with triethylamine and a solution of 3.7 g of benzyl 2-acetamido-3-0-carboxymethyl-2-deoxy-a-D-glucopyranoside and 1.38 ml of triethylamine in 100 ml of tetrahydrofurane is added.

After adding 2.6 g of 2-ethoxy-N-ethoxycarbonyl-l,2-dihydroquinoline (EEDQ), the mixture is left to stand foi1 24 hours at room temperature and is evaporated to dryness. The residue is dissolved in chloroform/methanol, 9/1, and this solution is washed with water, ice-cold 2 N hydrochloric acid, water, a saturated solution of sodium bicarbonate and water and the solvent is evaporated. The benzyl ester of benzyl-2-acetamido- 54 4 419 ο -C—ί t’L-l-(E-l~carb-jKeyl-J-carbpxj’propyli-carbamcyl-vi 'ij ?. ?cs rbaiaoylcethylj -2-deoxy-a-D-glucopyrsnoside, which ic thus obtained, is recrystallised from ethanol; melting point 208212°, ·“ +77° -1° (N,N-dimethylformamide, c = 0.546).

After splitting off the two benzyl radicals by hydrogenolysis, as described in Example 2, 2-acetamido-3~O-[[L-l(D-l-carbamoyl-3-carboxypropyl)-carbamoylethyl]-oarbamoylmethyl] 2-dr.oxy-D-glucoce is obtained.

Sxsinple ;52 3.8 g of the dimethyl ester of benzyl-2-acetamido-3~0£rh“l-(D~l,3“dicarboxypropyl)-carbamoyletbyl]-carbainoylirefchylJ 2-deoxy-a-D-glucopyranoside are dissolved in 100 ml of methanol and hydrogenated under1 normal pressure and at room temperature in the presence of 5% strength palladium-on-charcoal. After the absorption of hydrogen has ceased, the catalyst is filtered off and the filtrate is evaporated to dryness. The residue is taken up in 70 ml of vzater and freeze-dried. The resulting foam is the dimethyl ester of 2-acetamido~3-0-^[L~l~(Dl,3-dicarboxypropyl)-carbamoylethyl!-carbamoylmethylj-2-deoxyD-giucose which has an [a]p° of +23° -1° (vzater, c = 0.814).

The starting material used can be prepared as follows: 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) are added to a solution of 12.9 g of benzyl-2-acetamido4,6-0-benzylidene™3~0-carboxymethyl-2-deoxy--a-n-glucopyranoside and 4.0 ml of triethylamine in 100 ml of Ν,Ν-dimethylformamide and the mixture is left to stand at room temperature for 20 - 55 44190 hours. The solvent is then evaporated} the residue is taken up in chloroform and this solution is washed with water, icecold 2 N hydrochloric acid, water, a saturated solution of sodium bicarbonate and water. After drying over magnesium sulphate, the solution is evaporated to dryness. The residue is extracted with warm ethanol .and the dimethyl ester of benzyl2-acetamido-4,6-O~benzylidene~3-O- £ [L-1-(D-1,3-dicarboxypropyl) carbamoylethyl]-carbamoylmethylj-2-deoxy-a-D-glucopyranoside, xvhich has not dissolved, is filtered off and dried; [α]βυ = +22° ±1° (chloroform, c =- 1.160). .6 g of the dimethyl ester of benzyl-2-acetamido4,6-O-benzylidene~3-O~ J* [L-l- (D-l, 3-dicarboxypropyl) -carbamoylethyl]-carbamoylmethylj-2-deoxy-a-D-glucopyranoside are dissolved in 420 ml of glacial acetic acid and 280 ml of water and the solution is warmed to 10°C. After stirring for 4 hours at this temperature, the reaction mixture is cooled and evaporated to dryness. A further three times 100 ml of water are added to the residue and the mixture is evaporated to dryness each time. The residue is then taken up in chlorofoim and this solution is washed with water, dried over magnesium sulphate and evaporated to dryness. This gives the dimethyl ester of benzyl-2-acetamido-3-0-£[L-l-(D-l,3-dicarboxypropyl)carbamoylethylJ-carbamoylmeihylJ-2-deoxy-a-D-glucopyranoside as a yellowish resin .with an [a]p° of +31° -1° (chloroform, c = 1,070).

Example 35 A solution of 6.1 g of the benzyl ester of benzyl-3-0- 56 - 441S0 j[L-l-(P-l-carbamoyl-3-carbcxypropyl)-carbamoyl ethylj-carbamoylmethyl]-2-deoxy~2-propionamido-a-D-glucopyranoside in 200 ml of 2/1 tetrahydrofurane/water is hydrogenated under normal pressure and at room temperature in the presence of 0.6 g of 5% strength palladium-on-charcoal. After the absorption of hydrogen has ceased, the catalyst is filtered off and the filtrate is evaporated. 150 ml of water are added to the residue and the mixture is hydrogenated in the presence of strength palladium-on-charcoal until no further hydrogen is taken up.

The catalyst is filtered off and the filtrate is lyophilised.

This gives 3-0- j[L-l-(D~l-carbamoyl-3-carboxypropyl)-carbamoylethyl]-carbamoylmethyi|-2-deoxy-2-propionamido~D-glucose.

The starting material used can be prepared as follows; 0.3 ml of methanesulphonic acid is added to a solution of 90 g of benzyl-2-acetamido-2-deoxy-a-D-glucopyranosJde in 900 ml of Ν,Ν-dimethylformamide, whilst stirring, with the exclusion of moisture and whilst cooling with ice water. A solution of 60 ml of isopropenyl methy], ether in 240 ml of Ν,Ν-dimethylformamide is then added dropwise in the course of one hour and the mixture is stirred at room temperature for a further two hours and rendered alkaline with triethylamine. - The solvent is distilled, the residue is taken up in ethyl acetate and this solution is washed with water, dried over magnesium sulphate and evaporated to dryness. The product, that is to say benzyl2-ncetamido--2-deoxy~4,6-0-isopropylidene-a-D-glucopyranoside is crystallised from ether; melting point 136-137°, [a]^° = +103° - 57 4419 0 ίΐ° (chloroform, c = 1.125). 52·5 g of benzyl-2-acetamido~2-deo3ty-4,6~0~isopropylidene-a-D-glucopyranoside are dissolved in a solution of 225 £ of potassium hydroxide in 750 ml of ethanol and 40 ml of distilled water and the solution is boiled under reflux for 4¾ hours. After cooling, the reaction mixture is concentrated to half its volume and poured onto ice. The mixture is extracted with chloroform end the organic phase is washed vzith water, dried over magnesium sulphate and evaporated to dryness. The residue, that is to say benzyl~2-amino-2-deoxy-4,6-0-isopropylidene-a-D-glucopyranoside, is crystallised from ether; melting point 145-146°C, = +117° -1° (chlorofonn, c = 1.295).

A solution of 16.0 g of potassium bicarbonate in 192 ml of distilled water is added to a solution of 24.7 g of benzy12-amino-2-deoxy-4,6-0~isopropylidene-a-D-glucopyranoside in 192 ml of chloroform and the mixture is cooled to 0°. 8.16 g of propionyl chloride are now added dropwise in the course of 20 minutes, whilst stirring, and the mixture is stirred at this temperature for a further 50 minutes. The organic phase is now separated off, washed with water, dried over magnesium sulphate and evaporated to dryness. The product, that is to say benzyl-2-deoxy-4,6-0-isopropylidene-2-propionamido-a-D-glucopyranoside, is crystallised from ethyl acetate/petroleum ether; melting point 121-122°, [α]^θ = +112° ±1° (chloroform, c = 0.977). 1.25 g of pract. sodium hydride (Fluka) are added to a solution of 9.1 g of benzyl-2-deoxy-4,6-0-isopropylidene-2- 58 44190 propionamido-oc-D-glucopyranoside in 90 ml of acetonitrile and the mixture is stirred fox' 2 hours at 40°. It is then cooled to -5 to ~10°C and 4.2 ml of ethyl bromoacetate are added.

After a fui'ther 20 minutes, 10 ml of ethanol are added and the reaction mixture is neutralised with glacial acetic acid and evaporated to dryness· · The residue is partitioned between ether and water and the ether solution is '/ashed wi.th water, dried over magnesium sulphate and evaporated. The product, that is to say the ethyl ester of benzyl-3-0-carboxymethyl-2deoxy-4,6-0-isoprOpylidene-2-propionamido-a-D-glucopyranoside» is crystallised from ether/petroleum ether; melting point 9495°, [σ]^° = +145° ±1° (chloroform, c = 1.218). 22.5 ml of 1 N sodium hydroxide solution are added to a solution of 6.8 g of the ethyl ester of benzyl--3~O-carboxymethyl2-d.eoxy-4,6-0-isoi)ropylidene-2-propionamido-a-D-glucopyranoside in 70 ml of methanol. After hydrolysis of the ester has ended, 7.5 ml of 1 N hydrochloric acid are added and the mixture is evaporated, to dryness. The product is dissolved in 50 ml of N,N-dimethylformamide and subjected to a condensation reaction with 15 mmols of L-alanine-D-isoglutamine benzyl estertrifluoroacetate in the presence of 3 «72 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ). The reaction mixture is then evaporated to dryness and the residue is taken up in chloroform. This solution is washed with water, ice-cold 2 N hydrochloric acid, water,.a saturated solution of sodium bicarbonate and water, dried over magnesium sulphate and evaporated to dryness. The product is crystallised from dilute ethanol; - 59 % 4 41 θ Ο melting point 177-80°, [a]p° = t71° -1° (chloroform, c = 1.047). 15 ml of 1 N hydrochloric acid are added to a solution of 8.1 g of the benzyl ester of benzyl-3-O-f[L“l-(P~l-carbamoyl3-carboxypropyl)-carbamoylethyl]-earbamoyImethylj-2-deoxy-4,65 0-isopropylidene-2-propionamido-a-D-glucopyranoside in 150 ml of methanol and the mixture is left to stand for one hour at room temperature. 15 ml of 1 N sodium hydroxide solution are now added and the mixture is evaporated to dryness. The resulting benzyl ester of benzyl-3-0-£[L-l-(D-l-carbamoyl~310 carboxypropyl)-carbamoyl-ethyl]-carbamoylmethylj-2-deoxy-2propionamido-a-D-glucopyranoside is crystallised from methanol/ water and dried; melting point 208-210°, [a]£° = +75° -1° (N,W-dimethylformamide, c = 1.120).

Example 34 3-0-^[L-l-(D-l~Carbamoyl-3-carboxypropyl)-carbamoylethyl] carbamoylmethyl} -2-deoxy-2-caprinoyl-amido-D-glucose is obtained as a white foam , by catalytic hydrogenation of the benzyl ester of benzyl-3-0-([L-l-(D-l-carbamoyl3-carboxypropyl)-carbamoylethyl]-carbamoylmethylj-2-deoxy-220 caprinoylamido-a-D-glucopyranoside in N,N-dimethylformamide in the presence of 5% strength palladium-on-charcoal.

The starting material used can be prepared as follows: Analogously to the procedure described in Example 35, 24.7 g of benzyl-2-amino-2-deoxy-4,6-0-isopropylidene-a-D25 glueopyranoside are reacted with 16.7 g of caprinoyl chloride and the reaction mixture is worked up. This gives benzyl-2-deoxy9 r » 4,6-0-isopropylidene-2-caprinoylamido-a-D-glucopyranoside which - 60 44190 has an laj?° of +81° -1° (chloroform, c », 1.109). 0,·'; g of pract. sodium hydride is added to a solution of 3«8 g of benzyl-2-dee;:y-4,6-Q-isopropy'J.idene-2-caiWinoylamido~ff~D~glucopyr0no3i.de in 40 ml of acetonitrile and the mixture is stirred for 2 hours at 40°« This .mixture is then cooled 'to -10°, 1.4 ml of ethyl bromoacetate are added and the mixture is stirred for a further hour at 0°. When the mixture is worked up as described in Example 33, this gives the ethyl ester of benzyl-3-0-carboxymethyl-4,6-0~isopropylidene-2-c.aprinoylamido~K~D-glucopyranoside as a yellow-brown oil with an [a]p° of +114° ±1° (chloroform, c = 1.242).

Analogously to Example 33, the ethyl ester is hydrolysed, the product is subjected to a condensation reaction with Lalanine-D-isoglutamine benzyl ester-trifluoroacetate in the presence of EEDQ and the isopropylidene-ketal is split by mild acid hydrolysis. In this way the benzyl ester of benzyl-30-£[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoylethyl]~ carbamoylmethyl^-2~deoxy~2-caprinoylamido-a~D-glucopyranoside is obtained as a yellowish resin.

A 5% strength solution of benzyl-2~acetamido~2-deoxy-30·- £[L~l-(D~l,3-bis-methylcarbamoylpropyl)-carbamoylethyl]carbamoylmethylj -α-D-glucopyranoside in 1/1 distilled water/ methanol is hydrogenated using 5% strength palladium-oncharcoal, tlie catalyst is filtered off and the filtrate is evaporated. The product, that is to' say 2-acetamido-2~ deoxy-3-0-^ fL-l-(g-l,3-bis-methylcarbamoylpropyl)-carbamoyl- 61 4410') ethyl]-carbamoylmethylJ -D-glucose is freeze-dried; [a]^° The starting material can be prepared as follows: ·δ g of L-alanyl-D-glutamic acid bis-methylamidehydrochloride and 5-1 g of 2-ethoxy~N-ethoxycarbonyl-l,2dihydroquinoline are added to a solution of 9.1 g of benzyl-2acetamido-4,6-0-benzylidene-3-0-carboxymethyl~2-deoxy-a~Dglucopyranoside in 100 ml of Ν,Ν-dimethylformamide and 2.77 ml of triethylamine and the mixture is left to stand for /(8 hours at room temperature. The solvent is now distilled off, water is added to the oily residue and the insoluble matter 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-4,6-0-benzylidene-2-deoxy-3-0~£[L-l(D-l,3-bis-methylcarbamoylpropyl)-carbamoylethyl]-carbamoylmethylj -a-D-glucopyranoside.

By means of mild acid hydrolysis, as described in Example 29, with 60% strength acetic acid, the benzylidene group is split off and the product, that is to say benzyl-2ao etamido-2-deoxy-3-0-£[L-l-(D-l,3-bis-methylcarbamoylpropyl)carbamoylethylj-carbamoylmethylj-α-D-glucopyranoside is isolated.

Example 36 A 5% strength aqueous solution of benzyl-2-acetamido5-0-[L—l—(D-l-carbamoyl-3-carboxypropyl)-carbamoyl-2-hydroxyethylj-carbamoylmethyl]-2-deoxy-a-D-glucopyranoside is hydrogenated in the presence of Jj# strength palladium-on-charcoal, - 62 * 4 1 8 υ the mixture is filtered and tho filtrate is freeze-dried.

This gives 2-acctamido-J-O-J [L-l-(D-l-carbamcyl-3-carboxypropyl) carbaiaoyl-2-hydroxyetisyl ]-carbamoylmetliyl< -2-deoxy-D-glucobs.

The starting material can he prepared as follows: 3.26 g of L-serine-D-isoglutamine tert.-butyl esterhydrochloride and. 2.6 g of 2-ethoxy~N-ethoxycarbonyl~l,2~ dihydroquinoline (EEDQ) are added to a solution of 3.7 g of benzyl-2-acetamido~3~0-carboxymethyi-2-deoxy~o:~D-glucopyranoside and 1.3S ml of triethylamine in I50 ml of tetrahydrofurane and the mixture ia left to stand for 20 hours at room temperature. After evaporating off the solvent, the residue is dissolved in S/l ch] oiOform/methanol, tho solution is washed with water, ice-cold 2 N hydrochloric acid, water, a saturated solution of sodium bicarbonate and water and filtered and the filtrate is freed from the solvent. This gives the tert.-butyl ester of benzyl-2-acetam.ido-3-0-^ LL-l--(D-l-carbamoyl~3“Carboxypropyl)carbamoyl-2-hydroxy-ethylJ-carbamoylmethylj -2-deoxy-a-Dglucopyrano side.

By means of mild acid hydrolysis, as described in Example 11, the tert.-butyl ester is hydrolysed and benzyl-2-acetamido3-0-^[L-l~(D-l-carbamoyl-3-carboxypropyl)-carbamoyl-2-hydroxy~ ethyl]-carbamcylinethylj -2-deoxy-α-D-glucopyranoside is obtained. Example 37 A 5% strength solution of ben3yl-2-acetamido~3-0-^[L~ 1-(D-l-carbamoyl-3-carboxypropyl)-carbamoylbutyl]-ca rbamoylmethylj-2-dsoxy-ff-D-glucopyranoside in 1/]. methanol/water is - 63 44190 hydrogenated in the presence of 5% strength palladium-oncharcoal, the mixture is filtered and the filtrate is evaporated. The residue is dissolved in distilled water and freeze-dried. This gives 2-acetamido-3-O~£[L-l-(D~l~ carbamoyl-3-carboxypropyl)-carbamoylbutyl3-carbamoylmethyl| 2-deoxy-D-glucose.

The starting material can be prepared as follows: 3.28 g of L-norvaline-D-isoglutamine tert.-butyl esterhydrochloride ahd 2.6 g of 2-ethoxy-N-ethoxycarbonyl-l,2dihydroquinoline (EEDQ) are added to a solution of 3-7 g of benzyl-2-acetamido-3-0-carboxymethyl~2-deoxy-a~D-glucopyranoside and 1.38 ml of triethylamine in 100 ml of tetrahydrofurane and the mixture is left to stand for 24 hours at room temperature. The reaction mixture is worked up as in Example 38. This gives the tert.-butyl ester of benzyl~2-acetamido-3~0-£[L-l(D-l-carbamoyl-3-carboxypropyl)-carbamoyl-butylJ-carbamoylmethyl}-2-deoxy-a-D-glucopyranoside.

The tert.-butyl ester is split by mild acid hydrolysis. Example 38 A solution of 8.3 g of the benzyl ester of benzyl-30-£[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoylethyl]-carbamoylmethyl}-2-deoxy-2-stearoylamido-a-D-glucopyranoside in 200 ml of glacial acetic acid is hydrogenated under normal pressure and at room temperature in the presence of 5% strength palladium-on-charcoal. After working up, 3-0-^[L-l-(D-lcarbamoyl-3-carboxypropyl)-carbamoylethylJ-carbamoylmethyl} -2deoxy-2~stearoylamido-D-glucose is obtained as a white powder.

The starting material used can be prepared as follows: 0.75 S of pract. sodium hydride is added to a solution of 6.2 g of benzyl-4,6-O-benzylidsne-2-deoxy-2-3tearcylaffiidoa-D-gluoopyranoside in 150 al of Ν,Ν-dimothylformamide and the mixture is stirred at 40° for 1-J hours. The mixture is now cooled to -5°, 2.2 ml of ethyl bromoacetate are added and the mixture is stirred for· 4 hours at room temperaturo.

After adding 10 ml of glacial acetic acid and 400 ml of distilled water, the mixture is extracted with chloroform and the organic phase is washed with water, dried and evaporated to dryness.

The resulting ethyl ester of benzyl-4,6-O-benzylidene-3-Ocarbcxymethyl-2-deoxy-2-stearoylamido-a-D-glucopyranoside is crystallised from ethanol; melting point 151-154°, ~ •:-94° il° (chloroform, c = 1.186). 50.1 ml of 1 N sodium hydroxide solution are added to a solution of 23.4 g of the ethyl ester cf benzyl-4,6-0-bsnzylidene-3-0-carboxymethyl-2-deoxy-2-stearoylamido-tt-D-£lucopyranoside in 334 ml of methanol, and 334 ml of tetrahydrofurane and the mixture is stirred for 90 minutes at 50° and evaporated. The residue is stirred in ice water and the product is filtered off, washed with water, dried and recrystallised from ethanol. Sodium benzyl-4,6-0-benzylidene-3-0-carboxymethyl-2“deoxy-2stearoylamido-a-D-glucopyranoside.lHgO melts at 225-242° (with decomposition), -+4.5° ±1° (chloroform, c = 1.097)· mmols of L-alanine-D-isoglutamine benzyl estertrifluoroacetate and 4.95 g (20 mmols) of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) are added to a solution - 65 44190 of 14.4 g (20 mmols) of sodium benzyl-4,6-0-ben2ylidene-3-0·carboxymethyl-2-deoxy~2-stearoylamido~a-B~glucopyranoside monohydrate in 120 m3, of tetrahydrofurane and the mixture is left to stand for 20 hours at room temperature and evaporated . to dryness. The residue is dissolved in chloroform and this solution is washed with vzater, ice-cold 2 N hydrochloric acid, water, a saturated solution of sodium bicarbonate and water, dried over magnesium sulphate and evaporated. The product, that is to say the benzyl ester of benzyl-4,6-010 benzylidene-3-0-^[L-l-(D-l-carbamoyl-3~carboxypropyl)-C£rbaffloylethyl]-carbamoylmethylj -2-deoxy-2-stearoylamido-D-glucopyranoside is recrystallised from ethanol: = +62° £l° (Ν,Νdimethylformamide, c = 1.148).

Mild acid hydrolysis of this benzylidene derivative in 65% strength acetic acid leads to the benzyl ester of benzyl3~0-^[L-l-(D-l-carbamoyl~3“Carboxypropyl)-carbamoyletbyl3carbamoylmethylj -2-deoxy-2-stearoylamldo-a~D-glucopyrahoside; melting point 188-189°, [α]^0 =+59° ±1° (Ν,Ν-dimethylformamide, c = 1.022).

Example 39 A 5% strength aqueous solution of benzyl-2-acetamido-30-^[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoyl -2methyl propyl] — carbamoylmethylj-2-deoxy-a-D-glucopyranoside is hydrogenated in the presence of 5% strength palladium-on25 charcoal and filtered and the filtrate is lyophilised. This gives 2-acetamido-3-0-] [L-l-D-l-earbamoyl-3-carboxypropyl)-carbamoyl-2-methyl-propyl]-carbamoylmethylj -2-deoxy-D-glucose.

The starting mafcei’ial can be prepared as follows: 3-38 g of L-valine-D-isoglutamine tert.-butyl esterhydrochloride and 2.6 g of 2-ethoxy-N-ethoxycarbonyl-l,2dihydroquinoline are added to a solution of 3«7 g of benzyl—2acetarnido-3-0-carboxyinethyl-2-deoxy-a-D-gIucopyranocide and 1.33 m'l of triethylamine in 100 ml of tetrahydrofurane and the mixture is left to stand for 24 hours at room temperature and worked up as described in Example 38. This gives the tert.-butyl ester of benzyl-2-acetamido-3-0-^[L-l-(D-l-carbamoyl-3carboxypropyl) -carbamoyl-2 -methylpropyl ]-carbamoylmethylj -2deoxy-a-D-glucopyranoside, which is hydrolysed under mild acid conditions to give benzyl-2-acetamido~3-O“^rL-l-(D-l“Carbam0yl“ 3-carboxypropyl)-carbamoyl-2-methylpi’opyl]-carbamoylmethylj -2deoxy-c—D-glucopyranoside.

Example 40 A 5% strength solution of benzyl-2-acetamido--3-0-^[Ll-(D-l-carbamoyl-3-carboxypropyl)-carbamoyl-3-methylbutyl]-· carba oylmethylj-2-dcoxy-K-D-glucopyranoside in 50% strength aqueous methanol is hydrogenated in the presence of 5% palladium-oncharcoal and the mixture is filtered and the filtrate is evaporated. The residue is dissolved in distilled water, the solution is filtered again and the filtrate is freeze-dried.

This gives 2-acetamido-3-0-£[L-l-(D-l-carbamoyl-3-carboxypropyl]carbamoyl-3 - methylbutylj-carbamoylmethylj -2-deoxy~D-gluoose.

The starting material used can be prepared as follows: 3·52 g of L-leucine-D-isoglutamine tert.-butyl esterhydrochloride and 2.6 g of 2-ethoxy-N-ethoxycarbonyl-l,2- 67 dihydroquinoline (EEDO) are added to a solution of 3*7 g of benzyl-2-acetamido-3-o-carboxj>methyl-2-deoxy-K-D-glucopyranoside and 1.58 ml of triethylamine in 100 ml of tetrahydrofurane and the mixture is left to stand for 24 hours at room temperature and worked up as described in Example 38. This gives the tert.-butyl ester of henzyl-2-acetamido-3-0-^[L-l-(D-lcarbamoyl-3-carboxypropyl)-carbamoyl-3~methylbutyl]-carbamoylmethyl] -2-deoxy-a-D-glucopyranoside, and mild acid hydrolysis of this leads to benzyl-2-acetamido-3-0-^[L-l-(D~l-carbamoyl3-carboxypropyl)-carbamoyl-3-methylbutyl]-carbamoylmethylj -2deoxy-a-D-g]ucopyranoside.

Example 4l A 5% strength solution of benzyl-2-acetamido-3-0“^[Ll-(D-l~carhamoyl-3-carhoxypropyl)-carbamoylpropyl]-carbamoylmethylj-2-deoxy-a-D-glucopyranoside in 50% strength aqueous methanol is hydrogenated under normal pressure and at room temperature in the presence of 5% strength palladium-oncharcoal, the catalyst is filtered off and the filtrate is evaporated’. The residue is dissolved in distilled water and the solution is freeze-dried. This gives 2-acetamido3-0-[L-l-(D-l-carbamoyl-3-carhoxypropyl)-carbamoylpropyl]carbamoylmethylj-2-deoxy-D-glucose.

The starting material can be prepared as follows: 3.24 g of Ν-α-L-aminobutyryl-D-isoglutamine tert.-butyl ester-hydrochloride and 2.6 g of 2-ethoxy~N-ethoxycarbonyl1,2-dihydroquinoline are added to a solution of 3.7 g of benzyl-2-acetamido-3-0-oarboxymethyl-2-deoxy-a-D-glucopyrano68 4419ο side and 1.38 ml of tristhylamine in 100 ml of tetrahydrofux-ane and af ter the mixture lias stood for 2.4 hours at room temperature it is worked up as described in Example 36. The resulting tert.-butyl ester of benzyl-2-acetamido~2~0-^[L-l5 · (D-l-carbaraoyl-3-carboxypropyl)-carbamoylpropyl]-carbamoylmethyl}-2-deoxy-a-D-glucopyranoside is then hydrolysed under mild acid conditions to give benzyl-2~acetamido-3-0-{[L-l(D~l~carbamoyl-3-carboxypropyl)-carbamoylpropylj-carbamoylmethylj-2-deoxy-a-D-glucopyranoside· A 5% strength solution of benzyl-2-acetamido-3~0-^[Ll-(D-l-oaxlXimoyl-3-carboxypropyl)-carbcinoylphenyl-methyl]carbamoylmeihylj-2-deoxy-a-D-glucopyi'anoside in 50% strength aqueous methanol is hydrogenated under normal pressux'e and at room temperature in the presence of 5% strength palladium-on charcoal, the catalyst is filtered off and the filtrate is evaporated. The residue is dissolved in distilled water and the solution is freeze-dried and the residue is 2-acetamido3-0-^[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoylphenyl20 methyl ]-carbamoy3metbyl} -2-deoxy-D-glucose.

The starting material can be prepared as follows: 3.72 g of 1-phenylglycine-D-isoglutamine tert.-butyl ester-hydrochloride and 2.6 g of 2-ethoxy-N-ethoxycarbonyl1,2-dihydroquinoline (EEDQ) are added to a solution of 3·? g 25 of benzyl-2~acetamido-3-0-carboxymethyl-2-deoxy-o--D~glucopyranosido and 1.38 ml of triethylamine in 100 ml of tetrahydrofurane and the mixture is left to stand at room temperature for 24 - 69 5 44ΐθθ hours and worked up as described in Example 36. ' This gives tlie tert.-butyl ester of benzyl-2-acetamido-3-0-|[1-1-(11-3carbamoyl-3-carboxypropyl)-carbamoyl-phenylmethyl]~carbamoylmethylj-2-deoxy-a-D-glucopyranoside, which is hydrolysed under mild acid conditions to give benzyl-2-acetamido-3~O~^[L-l(D-l-carbamoyl-3-carboxypropyl)-carbamoylphenylmethyl]carbamoylmethylj -2-deoxy-a-D-glucopyranoside.

Example 43 ml of distilled water are added dropwise in the course of 15 minutes to a solution of 3*5 g oi benzyl-2-acetamido-30-£[L—1—(D—1,3-bis-carbamoylpropyl)-carbamoylethylj-carbamoylmethylj-2--deoxy-4,6-0-isopropylidene-p-D~glucopyranoside in 50 ml of glacial acetic acid, at 50°, whilst stirring, and the mixture is stirred for 1 hour at this temperature. After cooling, a further 100 ml of distilled water are added and the solvent is evaporated under a waterpump vacuum. 20 ml of water are added to the residue and the .mixture is freeze-dried. This gives benzyl-2-acetamido-3-0-[[L—1-(0-1,3-bis~carbamoylpropyl)-carbamoylethyl]-carbamoylmethylj-2-deoxy-p-D-glucopyranoside which has an [α]ρθ of -44° ±1° (Ν,Ν-dimethylformamide, c — 0.989).

The starting material used can be prepared as follows: 0.1 ml of methanesulphonic acid is added to a solution of 30 g of benzyl-2-acetamido-2-deoxy-p-D-glucopyranoside in 300 ml of Ν,Ν-dimethylformamide, whilst stirring and with external cooling and the exclusion of moisture. A solution of 20 ml of isopropenyl methyl ether in 60 ml of N,N-dimethyl- 70 44190 formamide is now added di-opwise in th? course of 1 hour and the mixture is stirred at roo» temperatiwe for a further two hours and rendered alkaline with triethylamine. After evaporating off the solvent, benzyl-2-s.cetamido~2-deoxy-4,65 O-isopropylidene-p-D-glucopyranos.ide is crystallised from ethylacetate; malting point 194°, [a]^ -· -104° £l° (chloroform, c ~ 0.550). 4.95 g of pract, sodium hydride are added to a solution of 36.2 g of benzyl-2~acetamido--2-deoxy~4,6“0~isopropylidene10 β-D-glucopyranoside in 400 rnl of acetonitrile and 100 ml of Ν,Ν-dimethylformamide and the mixture is stirred for 2 hours at 40°. This mixture is now cooled to -10°C, 17.2 ml of ethyl bromoacetate are added and the mixture is stirred for 30 minutes at 0°. 40 ml of ethanol are now added and the mixture is neutralised with glacial acetic acid and evaporated tc diyness. The residue is partitioned between ether and distilled water and tha ether phase is washed with water, dried over magnesium sulphate and evaporated. The product, that is to say the ethyl ester of benzyl~2-acetamido~3-G2o carboxymethyl-2-deoxy-4,6-0-.isopropylidene-p-D-glucopyranoside is crystallised from ether; melting point 93~94°, [a]^° = -49° ±1° (CHClj, c = 1.001). 22.5 ml of 1 N sodium hydroxide solution are added to a solution of 6.56 g of the ethyl ester of benzyl-2-aoetamido-325 0-carbamoylmethyl~2~deoxy-4,6-0-isopropylidene~p-D-gluco~ pyranosidc in 70 ml of methanol. After the hydrolysis of the ester has ended, 7.5 ml of 1 N hydrochloric acid are added .^4419° and. the mixture is evaporated to dryness. Tne resulting sodium salt is dissolved in 50 ml of N,N-dimethylformamide and 3.7 g of L-alanyl-D-glutamic acid-diamide-hydrochloride and 3·72 g of 2-ethoxy-N-ethoxycarbonyl-l,2-dihydroquinoline (EEDQ) are added. After standing for 24 hours at room temperature, the mixture is evaporated to dryness, the residue is dissolved in water and the solution is extracted with methylene chloride and twice with n-butanol. The product which remains after distilling off the n-butanol is benzyl-2-acetamido-3-0-^( [l1- (D-l,3-bis-carbamoylpropyl)-carbamoylethyl]-carbamoylmethylj 2- deoxy-4,6-0-isopropylidene-p-D-glucopyranoside.

Example 44 ml of distilled water are added dropwise to a solution of 5.1 g of the dimethyl ester of benzyl-2-acetamido-30-£[L-1-(D-1,3-bis-carboxypropyl)-carbamoylethyl]-carbamoylmethylj -2-deoxy-4,6-0-isopropylidene-p~D-glucopyranoside in 120 ml of glacial acetic acid, at 50°, whilst stirring, and the mixture is stirred for one hour at this temperature. After evaporating off the acetic acid, the dimethyl ester of benzyl2-acetamido-3~0-}[L-l-(D-l,3-bis-carboxypropyl)-carbamoylethyl]carbamoylmethylj.-2-deoxy-p-D-glucopyranoside is obtained.

The starting material used can be prepared as follows; 6.56 g of the ethyl ester of benzyl-2-acetamido-3-0carboxymethyl-2-deoxy-4,6-0-isopropylidene-p-D-glucopyranoside are hydrolysed as described in Example 43 with sodium hydroxide solution and the reaction product is subjected to a condensation reaction with 4.24 g of L-alanyl-D-glutamic acid dimethyl - 72 4419ο ester-hydrochloride in tho presence of 3.72 g of 2-et.rcxy-Nethoxycarbonyl-l,2-dihydroquinoline. After evaporating off the solvent, the residue is taken up in chloroform and the solution is washed with water, ice-cold 2 N hydrochloric acid, S water, a saturated solution of sodium bicarbonate and water, dried over magnesium sulphate and evaporated to dryness. Th< residue is the dimethyl ester of benzyl-2-acetamido-3-0-^LL-l(D-l,3-bic-carboxypropyl)-carbamoylethyl]-carbamoylmethylj-2deoxy-4,S-O-isopropylideno-p-D-glucopyranoside. io Example 4.5 A 5% strength solution of the benzyl ester of benzyI-2acetamido-3-0-^[L-1-(D-l-carbamoylmethylcarbamoyl-3-ci? rboxypropyl)-carbamoylcthyl]-carbamoylmethylj-2-deoxy-a-D-glucopyranouide in 2/1 tetrahydrofurane/water is hydrogenated under normal pressure and at room temperature in the presence of 5% strength palladium-on-charcoal. After about 50% of the theoretical amount of hydrogen has been taken up, the catalyst is filtered off and the filtrate is evaporated. The residue is now dissolved in distilled water and further hydrogenated.

After the theoretical amount of hydrogen Isas been taken up, the catalyst is filtered off and the filtrate is freeze-dried.

This gives 2-acetamido-3-0-^[L-l-(D-l-carbamoylmethylcarbamoyl3-carboxypropyl)-carbamoylethyl]-carbamoylmethylj -2-deoxy-Dglucosc in the form of a white povzder.

The starting material can be prepared as follows: 4.8 g of N-tert.-butoxycarbonyl-L~alanine-D-glutamine (benzyl ester)-glycine-amide monohydrate are dissolved in a - 73 44190 mixture of 5 ml of trifluoroacetic acid and 5 ml of 1,2-dichloroetbane and the solution is left to stand for 16 hours at room temperature, with the exclusion of moisture. This solution is diluted with 50 ml of tetrahydrofurane, cooled in an ice hath and neutralised with triethylamine. After adding a solution of 3.7 g of benzyl-2-acetamido~3-carboxymethyl-2deoxy-a-D-glucopyranoside and 1*38 ml of triethylamine in 100 ral of tetrahydrofurane, 2.6 g of 2-ethoxy-N-ethoxycarhonyl-l,2dihydroquinoline are added to the whole and the mixture is left to stand for 24 hours at room temperature and worked up as described in 'Example 36· This gives the benzyl ester of benzyl2- acetamido-3-0-|[L-l-(D-l-carbamoylmethylcarbamoyl-3-carboxypropyl)-carbamoylethyl]-carbamoylmethylj-2-deoxy-K-D-glucopyranoside.

Example 46 A solution of 5.2 g of the benzyl ester of benzyl-4,60-benzylidene-3-0~ £[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoylethyl]-carbamoylmethylj -2~deoxy-2-p--tolylsulphonylamino-a~ D-glucopyranoside in 120 ml of 70% strength aqueous acetic acid is hydrogenated under normal pressure and at room temperature in the presence of palladium-on-charcoal. After the theoretical amount of hydrogen has been taken up, the catalyst is filtered off and the filtrate is evaporated to dryness. The residue is dissolved in 30 ral of distilled water and the solution is freeze-dried. This gives 3-0-^[L-l-(D-l-carbamoyi3- carboxypropyl)-carbamoylethyl]-carbamoylmethylj -2™deoxy-2-ptolylsulphonylamino-D-glucose in the form of a white powder. - 74 44190 The starting material U3ed can be prepared as follows: A solution of 3.1 g of p-toluenesulphonyl chloride in 30 ml of methylene chloride is added dx-opvise, at 5°, to a solution of 6.0 g of benzyl-2-aHiino-4,6-0~benzylidene-3-0-carboxyroethyl-3“deoxy- This gives the benzyl ester of benzyl-4,6-0-benzylidene~3-0£[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoylethyl]-carbamoylmethylj-2-deoxy-2-tolylsulphon.ylamino-a~D-glucopyranoside in the form of a white powder. - 75 4419° Example 47 2-(Acetaminon!ethylcarbonylamino)-2-desoxy-3,0-^[L-]-(D·· l-c3rbamoyl-3-carhoxy-}.ropyI)-l-eur barney 1-ethyIS-cm. barneyi ~ methyl;-a,β-D-glucose is obtained analogously to Example 33.

Example 48 In a manner analogous to that described before 2-trimethylacetamido-2”desoxy-3,0- ([L-1-(D-l-carbamoyl-3-carboxypropyl)-1-carbamoyl-ethyl]-carbamoylmethy1>-α,β-D-glucose is obtained as Lyophilisate.

Claims (24)

1. A glucosamine derivative of the general formula (I) R, - CONH - CH(CH 0 ) 0 - R. (D) 2 2 and the salts thereof, in which X denotes a carbonyl or sulphonyl group, R denotes an optionally substituted alkyl radical or an optionally substituted aryl radical and, if X is the carbonyl group, R may also denote an alkoxy or benzyloxy radical, R^ denotes hydrogen, alkyl or an optionally substituted benzyl radical, R 2 denotes hydrogen or lower alkyl, R 4 and R g denote 10 hydrogen, alkyl or an optionally substituted benzyl or an acyl radical, R ? denotes hydrogen, alkyl, hydroxymethyl, mercaptomethyl or phenyl, βθ denotes an optionally esterified carboxyl group or amidised carboxyl group which is optionally substituted and Rg denotes an optionally esterified or amidised carboxyl 15 group which is optionally substituted, with the proviso (a) that the optionally substituted alkyl radical R has more than 1 carbon atom if (1) X denotes the carbonyl group and the radical R 2 denotes methyl, or (2), if X denotes the carbonyl group, the radical R 2 represents hydrogen and Rg and Rg each represent 20 a carboxyl group, and also with the proviso (b) that where R 2 and R 7 are other than hydrogen, the side-chain attached to oxygen in the 3-position of the glucosamine residue has the DLD configuration.

2. A glucosamine derivative of the formula (I), shown in Claim 1, in which X denotes a carbonyl radical and R denotes a lower alkyl radical which is optionally substituted by hydroxyl or carboxyl groups or denotes a phenyl radical which is option5 ally substituted by lower alkyl, lower alkoxy, trifluoromethyl or halogen, and its salts.

3. A glucosamine derivative according to one of Claims 1 or 2, In which Rg represents hydrogen. A. A glucosamine derivative of the general formula and the salts thereof in which R denotes lower alkyl or phenyl, R^ denotes hydrogen or lower alkyl, Rg denotes hydrogen or methyl, Ry denotes hydrogen, lower alkyl or hydroxymethyl, Rg denotes carbamoyl and Rg denotes carboxyl, with the proviso (a) that the lower alkyl radical R contains more than 1 carbon atom if Rg denotes methyl, and (b) that where R 2 and R ? are other than hydrogen, the side chain attached to oxygen in the 3-position of the glucosamine residue has the DLD configuration. - 78 44190

4. 5· A glucosamine derivative of the formula II, according to Claim 4, in which represents hydrogen, and R? represents hydrogen, methyl, or hydroxymethyl and its salts.

5. 6. Benzyl-3-0-^D-l-[L-l-(D-l-carbamoyl-3-carboxypropyl)5 carbamoyl-ethyl]-carbamoyl-ethylj-2-deoxy-2-propionylaminoα-D-glucopyranoside,

6. 7. A glucosamine derivative of the formula I, according to Claim 1, in which R^ and R 2 represent hydrogen, R denotes phenyl, Ry denotes methyl and Rg and Rg represent carboxyl, 10 carbomethoxy, carbamoyl or N-methyl-carbamoyl.

7. 8. A glucosamine derivative of thfe formula II according to Claim 4, in which R-j^ and R 2 represent hydrogen, R denotes methyl or phenyl, Ry denotes hydroxymethyl, Rg is carbamoyl and Rg carboxyl. 15

8. 9. A glucosamine derivative of the formula ll, according to Claim 4, in which R^ and R 2 represent hydrogen, R denotes phenyl, Ry is methyl, Rg represents carboxyl and Rg N-propylcarbamoyl or N-carbamoylmethylcarbamoyl.

9. 10. A glucosamine derivative, according to Claim 1, in vhich 20 R^ and R 2 represent hydrogen, R and Ry denote methyl and Rg and Rg represent carbomethoxy, carbamoyl or N-methylcarbonyl.

10. 11. A glucosamine derivative, according to claim 1, in which R represents phenyl, R^ denotes ethyl, R 2 represents hydrogen, Ry is methyl, Rg represents carbamoyl and Rg carboxyl or Rg 25 and Rg represent carbomethoxy or N-methylcarbamoyl.

11. 12. A glucosamine derivative, acoording to Claim 1, wherein R and Ry represent methyl, R^ is benzyl, Rg represent hydrogen, and Rg and R g represent carbomethoxy or carbamoyl.

12. 13. Benzyl-2-acetamido-3-0-[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoyl-ethyl]-carbamoyl-methyl-2-deoxy-6-0-stearoyla-D-glucopyranoside.

13. 14. 2-Acetamido-3-0-^D-l-[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoyl-ethyl]-carbamoyl-propylj -2-deoxy-D-glucose.

14. 15. 2-Benzoylamino-3-0-^D-l-[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoyl-ethyl3-carbamoyl-ethylj -2-deoxy-a,β-Dglucose.

15. 16. 2-Benzoylamino-3-0-^D-l-[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoyl-ethyl]-carbamoyl-propyl^-2-deoxy-a,β-Dglucose.

16. 17’. 2-Behzoylamino-3-0- £[L-l-(D-l-carbamoyl-3-carboxypropyl)-l-carbamoyl-ethyl]-carbamoyl-methyl^-2-deoxy-a,β-Dglucose·

17. 18. A pharmaceutical formulation containing one of.the compounds claimed in Claims 1 to 17. together with a pharmaceutical excipient.

18. 19. A pharmaceutical formulation containing one of the compounds claimed in Claims 1 to 17 together with a vaccine and, optionally, a pharmaceutical excipient. 8o 44190

19. 20. Process for the manufacture of a glucosamine derivative of the general formula X as defined in claim 1, wherein:(a) a compound of the formula III (III) R 2 “ CH :ooh in which X, R, and R 2 are as defined in claim 1, R^ 0 , R 4 °, and R g ° represent the radicals R^, R 4 , and R g respectively or represent a protective group, is condensed with a compound of the formula IV 10 H 2 N - CH - CONH - CH(CH 2 ) 2 - R g ° (IV) in which R ? o , Rg 0 , and Rg° possess the meanings of R ? , R g , and Rg, with the proviso that carboxyl groups, and if desired, free hydroxyl groups present in these radicals are protected by protective groups, and protective groups which are present 15 are split off, or (b) a compound of the formula V CONH - CH - COOH is condensed with a compound of the formula VI h 2 n CH - (CH 2 ) 2 (VI) and any protective groups which may be present are split off, or (c) a compound of the formula VII (VII) Is reacted with a compound of the formula VIII «7 K R I I 8 Z - CH - CONHCH - CONH - CH - (CH 2 ) 2 - R g ° (VIII) 10 in which Z represents a reactively esterified hydroxyl group, and any protective groups which may be present are split off, or (4) compound of the oxazoline and dioxolane rings formula IX in a (IX) (ch 2 ) 2 in which Rg represents an alkylidene or cycloalkylidene radical are split open under acid conditions with formation of the glucosamine ring and any protective groups which may be present are split off, and, if the radical R-COhas been removed, introducing the -X-R radical into the amino group in the 2-position of the sugar molecule, and, if desired, a compound resulting from any of the above processes is converted into a salt.

20. 21. Process according to claim 20 (c), wherein the acid of the formula III is reacted, in the form of the activated carboxylic acid, with the amino compound IV or the acid III is reacted with the compound IV in which the amino group is present in an activated form.

21. 22. Process according to claim 20 (d), wherein the acid of the formula V is reacted, in the form of the activated carboxylic acid, with the amino compound VI, or the acid V is reacted with the compound VI, in which the amino 5 group is present in activated form.

22. 23. Process according to claims 20 - 22, wherein the compounds of claims 2 to 17 are manufactured.

23.

24. Processes for the preparation, of compounds of formula I as defined in claim 1, as defined in claims 20 10 to 23, with reference to examples 1 to 48.