CA1262400A - Process for the manufacture of glucosamine derivatives - Google Patents

Abstract

PROCESS FOR THE MANUFACTURE OF GLUCOSAMINE DERIVATIVES
Abstract of the Disclosure A glucosamine derivative of the general formula (I) in which X denotes a carbonyl or sulphonyl group, R denotes an optionally substituted alkyl radical or an optionally substi-tuted carbocyclic aryl radical and, if X is the carbonyl group, also denotes an alkoxy or benzyloxy radical, R1 denotes hydro-gen, alkyl or an optionally substituted benzyl radical, R2 denotes hydrogen or Lower alkyl, R4 and R6 denote hydrogen, alkyl or an optionally substituted benzyl or an acyl radical, R7 denotes hydrogen, alkyl, hydroxymethyl, mercaptomethyl or phenyl, R8 denotes an optionally esterified or amidised carboxyl group and R9 denotes an optionally esterified or amidised carboxyl group, with the proviso that the optionally substituted alkyl radical R has more than 1 carbon atom if X denotes the carbonyl group and the radical R2 denotes methyl; or, if X denotes the carbonyl group, the radical R2 represents hydrogen and R8 and R9 each represent a carboxyl group, and its salts.
They have immunity-boosting action.

Description

The invention rel.ates to new glucosamine derivatives, i.e. glucosarnino-3-alkanoyl-dipep-tides of the general formula I

CH20?~6 f~) > ~ ~1 (I) / NK
/
R~

(D~
CO - MH - CH - CO~H ~ CH(CH2)2 ~9 in which X denotes a carbonyl or sulphonyl group, R denotes lower alkyl, lower alkyl substituted by hydroxyl or carboxyl, phenyl, lower alkyl phenyl, lower alkoxy phenyl, halophenyl or tri1uoro-methylphenyl, Rl denotes lower alkyl or benzyl/ R2 denotes hy-drogen or lower alkyl, R~ and R6 denote hydrogen, alkyl or alka-noyl, R7 denotes hydrogen, lower alkyl, hydroxymethyl, mercapto-methyl or phenyl, R8 denotes carboxyl, lower alkyl esterifi?d carboxyl, carbamoyl, lower alkyl carbamoyl, ben~yl carbamoyl or carbamoylmethyl carbamoyl, and Rg denotes carboxyl, lower alkyl esterified carboxyl, benzyl esterified carboxyl, carbamoyl and lower alkyl carbamoyl, with the proviso that the optionally substituted alkyl radical R has more than one carbon atom if X

denotes a carbonyl group and the radical R2 denotes methyl, or, if X denotes a carbonyl group, the radical R2 represents hydrogen and R8 and Rg each represent a carboxyl group, and their salts.

Unless specifically stated) residues~ radicals or com-pounds modified by -the -term "lower" in -the text which follows con-tain preferably up to 7 and above all up to 4 carbon atoms.
Alkyl is5 in particular, lower alkyl, for example iso~
propyl, straight-chain or branched butyl, pentyl, hexyl or heptyl, ~lich are bonded in any desired posi.-tion, and above all methyl, ethyl or n-propyl.

The substituted alkyl radical, such as a lower alkyl radical, can carry one, two or more identical or different sub-stituents, especially free hydroxyl groups or halogen atoms.

Alkanoyl has preferably 2-18 carbon atoms and i5 e.g. acetyl or propior,yl.

As a substituent of ~the abovemen-tioned radicals, lower alkyl is above all methyl or ethyl but also n-propyl, iso-propyl or straigh-t-chain or branched butyl.
As a substituent of the above~entioned radicals, lower alkoxy is, in particular, methoxy or ethoxy and also n-propoxy, isopropoxy, n-butoxy or isobutoxy.

Halogen is, for example, fluorine, chlorine or bromine.
Carbo-lower alkoxy groups which may be mentioned in particular are carbomethoxy or carboe-thoxy, but also carbo-n-propoxy or carbo-isopropoxy groups, and carbamoyl groups which may be mentioned in particular are the carbamoyl group itself.
In the abovementioned compounds, in which R2 denotes an alkyl radical, the R2-acetamide radical which is linked to the oxygen ato~ in the 3-position of the glucosamine radical is optically active, that is to say it is in the D-form. If R7 does not represent hydrogen, the R7-aminoace-tic acid is in the L-form.

f 3~ f~

Depending on the nature of their substituents, the present new compounds are neutral or acid 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.
Salts are, in particular, non-toxic salts which can be used pharmaceutically. Other salts 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 characterisation.
The cornpounds of the present inventior display valuable pharmacological properties, in particular a marked immunity-boosting action. This can be demonstra-ted with the aid of the test arrangement described below:

1. Boosting of the cellular imrnuni-ty in ViVG: increase in the dela~ed--tyr~e hlpersensitivity to ovalbumin in ~uinea r~iF~s Pirbright guinea pigs are immunised on day O with 10 rng of ovalbumin in complete Freund's adjuvant by injecting O.l ml of an antigen-adjuvant mixture into each of the two back paws.
4 weeks' later skin reactions are produced by intracutaneous injection OI 100 ~g of ovalbumin in O.l ml of buffered physio-logical 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 of cell-transmitted immunity.
Ovalbumin is too weak an immunogen to induce a delayed-type reaction on its own or in a water-in-oil emulsion with incom-plete Freund's adjuvant (10 parts of ovalburnin solution in 0.9%
of NaCl mixed with 8.5 parts of Bayol F and 1.5 parts of Arlacel A), but, for effective immunisation9 has to be administered in complete adjuvant, to which Mycobacteria are added (5 mg of ., .

M butyricum, which have been destroyed ancl lyophilised 9 per 10 ml of Bayol F/Arlacel A). In order to demonstra-te the immunity-boosting action OI the test subs-tances, the latter can now be mixed, in doses of 10 to 100 ~g, with the an~tigen/oil mixture in place of the Mycobacteria.
The glucosamine peptides according to the invention are able to imitate the effect of the Mycobacteria i~ 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 mix-ture but are administered subcutaneously~ in sodium chloride solution, in doses of 10 to 100 ug 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._ _ _ _ NMRI mice are imm~nised by intraperitoneal (i.p.) injec-tion of 10 ~g of precipitate-free BSA on day 0. Serum samples are taken 9, 15 and 29 days later and tests are carried out~
using a passive haemagglutination technique;to determine their content of an-ti-BSA antibodies. In the dosage used, soluble BSA i.s su~)-immunoLel-lic for ~the receivin.g animals, that is to say it is not able -to initia-te any production of antibodies or is able to ini-tia~te only a very slight production of an-tibodies Additional trea~tment of the mice with certain immuni-ty-boosting subs~tances before or after -the adminis-tration of the antigen leads to a rise in the antibody ~titre in the serumO The effect of ~he treatmen-t is expressed by the score value, tha~
is to say by the su.m of the log2 titre differences on the three days when blood is sampled, which is reached.
On in-traperitoneal or subcutaneous (s.c.) admi.nistra-tion of 100-300 mg/kg/animal on five consecutive days (day O to 4) after immunisation with BSA, compounds of the present inven-tion are able significantly to increase the production of anti-bodies against BSA.
The immunity-stimulating effect of the sai.d compounds, in contrast to that of other bacterial immunoleptic agents (for example LPS from E. coli), is dependent on the antigen: injec-tion of the new compounds results in an increase in the anti-BSA titre only in BSA-immunised mi.ce, but not in mice which have not been immunised. It is worthy of note ~that the s.c.
administration of the said compounds is equally as effective as the i.p. administration, that is to say the immunity-boosting action which is observed is systemic and does not depend on the s-timulant 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.
Tne experiments i~strated show that compounds of the type ~,~$~ g ~

described are âlsO able specifical~Ly -to increase -the humoral immurit~, -that -they improve the i~munological response and that their immunity-boosting effec~ts are based on a systemic ac-tiva-tion of the immune system.

3. Boosting of the humoral irnmunity in vitro: T~cell-replacemen-t effect in ~the antibody response of mice sple~ocytes to sheeP ~r~throc tes (SE).
In many cases, lyrnphocytes (T-cells) emanating from -the thymus are necessary for the induction of an antibody response~
m ese cells co-operate with the precursors of antibody-form:ing 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 frorn congeni-tally athymic nu/nu mice do not contain any func-tional T-cells and, for example in vitro in the presence of SE, are not able -to form any an-ti-SE an-tibodies. m e compounds of the present invention are, surprisingly, able functionally to replace T-cells in such cul-tures and to make an antibody response to SE possible. The addition ofthese substances to nu/nu splenocyte cultures in the presence of SE leads, within

4 days, to a considerable increase in the number of antibody-forming cells. me findings show that the said cornpounds are able -to increase the humoral antibody formation in vitro and -to compensa-te a defect in the T-cell system.
4. Selective mitogeneity for B-cells: proliferation-promotin~ effect in B-l~rnphocyte cultures. _ _ _ Suspensions of highly enriched B-lymphocy-tes (lymph n~e cells from congeni-tally athymic nu/nu mice) and sub-stantially ure immature and mature T-lymphocytes (thymocytes and, respectively, cortisone-resistant thymocytes, that is to say thymocytes which persist 48 hours after a cortisone injec-tion, from Balb/c mice) are incubated for three days in the presence OI thetest subs-tances. The incorporation of H3-thymidine in the lymphocytes during the final 18 hours of the culture period is taken as a measure of the prolifera-tion acti-vity.
The compounds of the inven-tion are mitogenic for B-lymphocytes (tha-t is to say for the precursors of the antibody-forming cells) but not for T-lymphocy-tes.
They are thus able to stimulate the proliferation of lymphocytes which are invo]ved in the humoral immune response.

5. Tolerance.
-Although compounds of the type described already dis-play their boosting action in guinea pigs after, for example, an individual dose of 0.05 mg/kg administered subcutaneously,and in mice a~rthesubcu-taneous 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 there-fore possess an excellent therapeutic range.
The compounds according to the invention are able, on the one hand, when mixed with an antigen7 to increase the immuno-genei-ty of the latter and, on the other hand, on systemic administration, to increase the immunological reactivi-ty of the trea-ted organismO The said substances are able to promote both the cellular and -the humoral immunity and to activate the lymphocytes .~hich are responsibie for the formation of anti-bodies.
rne new compounds can -thus be used, as adjuvants mixed with vaccines, to improve ~the success rate of the vaccina-tion and to improve the protection impar-ted by humoral antibodies and/or cellular immunity agains-t infection by bacterial, viral or parasi-tic pathogens.
Finally, the compounds described are suitableg as a mixture with very diverse antigens, as adjuvants for the experl-mental and industrial production of antisera for therapy and diagnosis and for inducing i~munologically 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 suit-able 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) immuno-logical defective states which are congenital or acquired, such as arise in old age, in the course of severe primary diseases and, above all, after therapy with ionising rays or with hor-mones having an immuno-suIpressive action. The said sub-stances can -thus be administered, preferably also in combination witln anti-infectious antibiotics, chemotherapeutic agents or other courses of treatment,in order to counteract immlmological damage. Finally, -the subs-tances described are also suitable for the general prophylaxis of infectious diseases in humans and animals.
The invention relateS in particular -to compounds of the formula (I) in which X denotes a carbonyl radical and R deno~tes a lower alkyl radical which is optionally substituted by hyclroxyl or carboxyl groups or denotes a phenyl radical l,lhich is option-ally subs-ti-tuted by lower alkyl, lower alkoxy, trifluoromethyl g , d R1, R2, R4, R6, R7, R8 and Rg have the above-mentioned meaning, and their salts.
Compounds in which R~ represents hydrogen and the other radicals possess the abovementioned meaning, and their salts are also particularly valuable.
Compounds to be singled out are, in particular, com-pounds of the general formula CH20}~ -~0 ~' ~\~-~1 ~H - CO - R (II) R - C - H

(D) \ IR7 lR8 CO - ~1 - CH - CO~ - CH - (CH2)2 9 (L) (D3 in which R denotes lower alkyl or phenyll Rl denotes ben~yl or lower alkyl, R2 denotes hydrogen or methyl, R7 denotes hydrogen~ lower alkyl or hydroxymethyl, R8 denotes carbamoyl and Rg denotes carboxyl, wlth the proviso that the lower alkyl radical R contains more than 1 carbon atom if R~
denotes methyl, and their salts.
Compounds to be mentioned above all are those of the formula II in which R denotes lower al~yl or phenyl, R
denotes benzyl or lower alkyl, R2 denotes hydrogen or methyl, R7 denotes hydrogen, rnethyl or hydroxymethy]., ~8 denotes carbamoyl and Rg denotes carboxyl, with the proviso that the lower alkyl radical R contains more than l carbon atom if R2 denotes methyl, and their salts.
~ he new compounds can be obtained when a compound of -the formula C~l2~6 4 ~ 0 - Rl (III) NH - X - R

R2 ~ CH

(D) COOI-I

in which X, R and R2 possess the abovementioned meaning and Rl 9 R4 and R6 represent the radicals Rl, R4 and R6 respec-tively or represent a protective group which can be split off easily, or a derivative thereof is subjected, in a manner rhich is in itself known, to a condensation reaction with a compound of the formula '~ ~' ?~ r~l R7e~ R8O ~ IV) O
C~ - Co.`~ C~ 2 ) ~- 9 (L) (D) in which R7, R~ and Rg possess -the meaning of ~7, R8 arld Rg, with the proviso that carboxyl groups and, lf desired, free hydroxyl groups present in -these radicals are protected by pro~
tective groups which can be split off easily, and any protective ~rollps which may be prese~t and ~o not belong -to the ~esired end product of the formula I are split o~f.
The condensation reaction is carried out, for example, by reacting the compound IIl 9 in ~the form of the activated car-boxylic acid, with the amino compound IV or by reacting the acid III with the compound IV, in which the amino group is present in an activated form. The activated carboxyl group can be for example, an acid anhydride, preferably a mixed acid anhydride, such as an acid azide, an acid amide9 such as an imidazolide, an isoxazolide or an activated ester. Activated esters which may be mentioned in particular are: cyanomethyl esters, carboxymethyl esters,p-nitrophenyl thio esters, p-nitrophenyl esters, 2,4,5-trichlorophenyl esters, pentachlorophenyl esters 9 N-hydroxy-succinimide esters, N-hydroxypthalimide esters, 8-hydroxyquino-line esters, 2-hydroxy-1,2-dihydro-1-carboe-thoxy-quinoline esters, N-hydroxypiperidine esters or enol esters which are obtained with N-ethyl-5--phenyl-isoxazolium 3'-sulphonate. Activated esters can also optionally be obtained with a carbodiimide with the addition of N-hydroxysuccinimide or a l-hydroxybenzotriazole or 3~hydroxy-4-oxo-3,4-dihydro-berzo~d]-1,2,3-triazine which is - 15 ~

~ J:~u unsubsti u~ed or substituted by, for examp]e, halogen, methyl or metho~
The arnirlo group is activated by, for example, reaction with a phosphite-arnide~
.I-~ongst -the me-thodscomprising a reac-tion with activated esters, those with N-ethyl-~-phenyl isoxazolium 3'-sulphonate (Woodward reagent K) or 2-ethoxy-1,2-dihydro-l-carboethoxy-qui.noline or carbodiimide are to be mentioned in p~rticular.
Protective groups which can be split off easily are those which are known from peptide chemistry and sugar chemistry.
Protec-tive 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 acetyl9 aroyl radicals such as benzoyl, and, above all, radicals derived from carbonic acid, such as benzyloxycarbonyl or lower alkoxycarbonyl, or alkyl, especially tert.-butyl, benzyl or tetrahydropyranyl which are optionally substitu-ted by nitro, lower alkoxy or halogen, or optional]y substi-tuted 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 optionall.y substituted and preferably substituted in the p-position.
These pro-tected groups can be split off in a manner which is in itself kno~m. Thus, ~they can be removed hydrogeno-lytically, for exarnple with hydrogen in the presence of a noble _ 16 _ r ~

metal c~-tal,--t, such as a palladium or pla-tinum catalys-t, or by acid hydrol.~-sis.
The startlng ma~erials used are known or can be manufac-tured in a mar~ler which is in i-tself :known. Thus, compounds of the formula III can be obtained, for example, by reacting the corresponding sugar, which is unsubstitu-ted in the 3-posi-tion, with a halogeno-R2-acetic acid, in which R2 has the abovementioned meaning, and its esters in th.e presence of a strong base. In these compounds J halogen is preferably bromine or above all chlorine.
Another procedure for the manufacture of the new glucos-amine derivatives is to subject a compound of the formula V

,: ~~0 4 _ O~ O - Rl (V) / ~H - ~ - R

(D)\ R7 co.n~ - CH - COOH
(L) in which R, Rl, R2, R4, R6 and R7 have the abovementioned meaning, in a manner which is in itself known~to a condensation reaction with a compound of the formula ~, o H2N - ~H - (CH2)2 - R9 (VI) (D) in which R8 and Rg have the abovementioned meaning, with the proviso tha~t carboxyl groups and, if desired,free hydroxyl groups present in the radicals R7, R8 and Rg are protected by pro-tective 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 reac-ting the compound V, in -the form of the activated car-boxylic acid, with the amino compound VI, or by reacting the acid V with the compound VI 5 in which -the amlno groups are in an activated form. The ac-tivated 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 par-ticular, 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 manner which is in itself known; for example hydrogenolytically, for example with hydrogen in the presence of a noble metal catalyst, such as a palladium or platinum catalyst, or by acid hydrolysis.
The starting materials can be obtained in a manner which is in itself known. Thus, it is possible, for example, to react corresponding sugars which are unsubstituted in the 3-position with a halogeno-R2-acetamido-R7~acetic acid, or to react a compound of the formula III in the manner indicated above with an amino-R7-acetic acid in which the carboxyl group - 18 _ is protected and to split off the protec-tive group.
The invention relates also to a process for -the manufacture o~ a glucosamine deri~ative of the general formula I
Cr~ 2 R 6 L--o ~,,,0 - Rl ` 0 / ( I ) NH - Y -- R

R 2 - C I Ij' 7 R ~3 CO ~ -- Cll -- CO~ Cll (CH2) 2 ~ R~3 (D) (L) (D) in which X denotes a carbonyl or sulphonyl group, R denotes lower alkyl, lower alkyl substituted by hydroxyl or carboxyl, phenyl, lower alky]. phenyl, lower alkoxy phenyl, halophenyl or trifluoro-methylphenyl, Rl denotes hydrogen, lower alkyl or benzyl, R2 denotes hydrogen or lower alkyl, R4 and R6 denote hydrogen, alkyl or alkanoyl, R7 denotes hydrogen, lower alkyl, hydroxymethyl, mercaptomethyl or phenyl, R8 denotes carboxyl, lower alkyl ester-ified carboxyl, carbamoyl, lower alkyl carbamoyl, benzyl carbamoyl or carbarnoylmethyl carbamoyl, and Rg denotes carboxyl, lower alkyl esterified carboxyl, benzyl esterified carboxyl, carbamoyl and lower alkyl carbamoyl, with the proviso that the optionally substituted alkyl radical R has more than one carbon atom if X
deno-tes a carbonyl group and the radical R2 denotes methyl, or, if X den.otes a carbonyl group, the radical R2 represents hydrogen and R8 and R9 each represent a carboxyl group, and its salts, wherein (a) a compound of the formula C~ O -- R
,~-0 ~ \ G - R (VII) R4 ~ ~
~H - X - P
i.n which X, R, P~l , R4 and R6 possess the abovementioned meanings of X, R, Rl, R4 and R6, provided that any hydroxyl groups which may be present -therein are protected by a protective group which can be split off easily, is reacted with a compound of the formula r~ R
17 18 (VIII) Z - CH - CONHCH - CONH - CH - (CH2)2 - R9 R2 (L) (D) in which Z represents a reactive esterifled hydroxyl group and R7 , R8 and R9 have the abovementioned meanings of R7, R8 and Rg provided that any protective groups which may be present therein are split off hydrogenolytically or by acid hydrolysis; or (b) for the manufacture of a compound of the formula I whereln R4 and R6 represent hydrogen, the oxazoline and dioxolane ring in a compound of the formula O

O \

,~ O / \~ (I'~) ,,,?<~1 1 N = C - R
(D) R C~ R7 R o CO~HCH - CONH -- C~i - (CH2)2 - Rg (L) (D) } ~

in ihich 1', R2, R7, 1~8 and Rg possess the abo~ementioned meanin~ and P~5 re2resents an alkylidene or cycloa]kylidene rad cal, is spli~ open under acid conditions and ~ny protective groups which ma~ be present are spl.it of~ hydrogenolytically or by acid hydrolvsis and the X-R radical is introduced into the amino group in the ~-position of the sugar molecule if this group has been liberated and, if desired, a resultir.g compound is con-verted into its sal-ts.
In the following, the ahove-mentioned process variants a) and b) are described in more detail:
Process a):
A reactively esterified hydroxyl group is, in particular, a hydroxyl group esterified with a strong inorganic or organic acid and above all a hydroxy] group which is esterified with hydrogen halide acids, such as hydrochloric acid, hydrobromic acid or hydriodic acid.
The protective groups which can be split off easily correspond to those already mentioned above. They can be split off in a manner which is in itself knowm, for example hydrogenolytically, for example with hydrogen in the presence of a noble metal catalyst, such as a palladium or platinum cata-lyst, or by acid hydrolysis.
The starting materials used in this process variant are kno~m.
Process b~:
In the compound of the formula IX, alkylidene is, in particular, lower alkylidene, such as isopropylidene, and cycloalkylidene is, above all, cyclopentylidene or cyclohexylidene.

~ 3~

This s?litting is also carried out in a ma~ne-r which is in i-tself kno~rn, for e~arnple with an acid ion exchanger, especi-ally those containing sulphonic acid grroups, such as ~berlite IR-120 (a styrene resin wi-th highly acid sulpho groups) or Dowex 50 (polystyrenesulphonic acids) or a strong inorganic or organic acid, such as hydrochloric acid, hydrobromic acid, sul-phuric acid or a sulphonic acid, for example methanesulphonic acid, or a phenylsulphonic acid which is optionally substituted in the aromatic ring, such as p-toluenesulphonic acid, or tri-fluoroacetic acid. If the reactiorl is carried out in the pre-sence of water a free hydroxyl group is obtained in the l-position;
if, on the other hand, the reaction is carried out in the pre-sence of an alcohol of the formula H0-Rl in which Rl represents lower alkyl or benzyl, the l-0-Rl compound is obtained. If one of the carboxyl groups R8 and/or Rg is also esterified with a lower alkanol, this can be saponi~ied with aqueous acid, especially at elevated temperature.
However, it is also possible for the amino group in the 2-position of the sugar molecule to be liberated during this splitting. In this case, the X-R radical has to be intro-duced subsequently. This is carried out in the customary manner by acylation or sulphonylation.

In the resulting compounds it is possible subsequently to split off protective groups on the peptide radical, for example by hydrogenolysis, such as, for example 9 wi-th catalyti-cally activated hydrogen, or by hydrolysis.
m e starting ma-terials used can be obtained, for example, when the R2-ace-tamidopeptide radical is introduced, in one or several stages 3 into a corresponding oxazoline which has a free ~ 3~
hydroxyl group in -the ~-posi-tion of -the sugar r~sidue.
The resulting compounds can be converted into -their salts in a manner which is in i~tself kno~, îor exarnple by reac--ting resul-ting acid compounds wi-th a:lkali me-tal hydro~.ides or alkaline earth metal hydroxides or by reacting resulting basic compounds with acids The processes described above are carried out in accor-dance with methods which are in themselves known, in the absence or, preferably, in the presence of diluents or solvents, if necessary wi-th cooling or warming, under eleva-ted pressure and/or in an inert gas atmosphere, such as a nitrogen atmosphere.
Ta~ing into account all of the substituents presen~t 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 suit~
able 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 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 pro cess, 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. The pharmaceutical fo-rmulations according to the invention are those for enteral, such as oral or rectal, and parenteral administra-tion to warm-blooded 2nimals and which contain ~the pharmacological active compound on its o~m or ~together wi-th an excipient which can be used pharmaceutically. The dosage of the active compound depends on the species of warm-blooded ani-mal, on the age and on the individual state and on the mode of administration.
The new pharmaceutical fo~mula-tions contain from about 10% to abou~t 95~6, and preferably from about 20% to about 90%, of the active compound. Pharmaceutical formulations accor-ding 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 manner which is in itself knol~m, for example by means of conventional mixing, granulating, dragee-making, dissolving or lyophilising processes. Apart from the types of administration mentioned on this page, pharmaceutical z4~

~ / 'S.o'r r)~ C;~ !,a~ 8.:1.so '~ b-t;~ *.~.r-J~
combi.nii~S, th~-- ^ct,,~.rr eOr,.po"-f!l Witl.l soli.~ e-~icipier.~.ts, optlo.~a.!:Ly gl~nl1la-t,ing a re,l.l]..~~-;ing r~.iY.ture and ~prcicessirlæ trl~ mix~t;urc or grOi-lules~ alter addir10 suit.3.ble aw~iliar:ies i.~ desired 5r .~'e~ s sary~ to gl~re -ta.b3ets or dr~ee cores.
Suitable e~.ci.p:ients arc, in par-tict!~l.ar. fil:Lerc.~ 5UC,"i as sugar~ for~ ar~lpl.e 1--ic~tc~s~ c,r s~!c~roC;~ rn~r~Lltol or ~;o~b:i-'ol, celLu'oie formt11a-tions a.~.~d/or calcll.lm phc~soh.ate~,for ex~rnpl.e t;rica:Lcit1~n phospha~te or calci~m 'rli:-drcgen ~hosphate, as well as b.inders, such ~s starch pastes u~,:ing, or ~xaL~Iple9 n-iaj.ze st?rch, ~heat s~varch, rice starch or pota~o starch, gelati.-~l5 traOacan-th~

nmet~lylcellulose, hyclro.~ ropy:lme~ lcellulo.se~ sc~clitlm carboxy~-methylcellulose and/or po'yvinylpyrro:lidoxle$ a-!~d/ors i~ desired~
disin-tegI~ating agents, suc.h as the abovementioned StarGheS 7 ancl also carboxymethyl-s-tarch~ crosslin~ed polyvi.nylPyrrolJ.done~
agar or alginic ac.id or a salt -thereof, such as sod:ium algina-te~
Auxiliaries are, above all, Mow~re~.la-ting agen-ts and l~abri--cants, .~or e~a~lple silica, talc ? stear.ic acid or sal-ts t,hereof, such as magnesiu~ s-tear.ite or ca].cil~ stearate~ and~or poly--ethylene glycol~ Dragee cores are pro~-ided ~,lth suit,able ccatings whichr if desired, are resistant to ~astri.c Juices5 ann For this pLl~pose~ inter alia, concen-trated suga-.. solutions, whic'h optionally cont:ain gum arab?c, talc, polyvinylpyrrolidonej poly~
ethylenQ glycol and/or ti+anium dio~ide, 1.ac~u&f solutions in suitable orga-rli.c solv~nts or solvent mi~tures or, in order to producecoatings resist~nt togastric jujces5 solutions oF suitab'!.e cellulose formula-ti.ons? such as ace-i:,ylcel]1l10se phthalate or hydro~-ypropylrnethylcellulose phthala-teJ are used. Dyestuffs or pigmen~ts can be added to the tablets or dragee coatings, for example for identification or in order -to characterise different doses of the active compound.
The e~amples which follow illus-trate the invention des-cribed above; they are not, however, intended to restrict the scope of the i~ven-tion in any way.Temperatures are given in degrees centi-grade.
E am~le 1 A solution of 2.6 g of the benzyl ester of benzyl-3-O~ - [L-l- (D-l-carbamoyl-3-carboxy~propyl)-carbamoyl-ethyl]-carbamoyl-ethyl~-2-deoxy-2-propionylamino-~-D-glucopyranoside in 67 ml of 60% strength ace~tic 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)0 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.
This gives benzyl-3-0-~_-1-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl~ carbamoyl-ethyl~-2-deoxy-2-propionylamino-~-D-glucopyranoside which has a melting point of 155-160 (decompo-sition) and an optical rotation [~]DO cf = +105 -1 (dimethyl-formamide, c = 0.5~).
The benzyl ester of benzyl-3-0-~D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyl-ethyl~-2-deoxy-2-propionylamino-~-D-glucopyranoside which is used as the starting material can be obtained as fol1ows:
4 1 g of benzyl-2-amino-4,6-0-benzylidene-3-0-(D-l-carboxye-thyl)-2-deoxy-~-D-glucopyranoside are dissolved in 200 ml of me-thanol and 3 ml of triethylamine and 0.95 ml of pro-pionyl chloride is added dropwise at room ~temperature, whilst stirring~ After stlrring 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 ice-cold 2N hydrochloric acid and the produc-t which has separa-ted out is filtered off, rinsed with water and dried in vacuo over sodium hydroxide. The benzyl-4,6-0-benzylidene-3-0-(D-l-carboxyethyl)-2-deoxy-2-propionylamino-a-D-glucopyranoside which is thus Gb ained can be recrystallised from methanol; melting point 257, [a~D = +132 1 (dimethylformamide, c = 1.086).
1.5 g of N-e-thyl-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 OI tri-ethylamine and the mixture is stirred at room temperature until everything has dissolved (about 60 minutes). 2.2 g of L-alanyl-D-glutamic acid l-amide-~-benzyl 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 thor-oughly with water and dried. The resulting benz-yl ester of benzyl-4,6-0-benzylidene-3-0- ~-1-[L-1-(D-1-carbamoyl-3-carboxy-propyl) carbamoyl-ethyl~-carbamoyl-ethyl~-2-deox-y-2-propionylamino-- 26 _ ~-D-glucopyranoside, which has an Rf value Gf 0.45 on silica gel thirl layer plates in ~the system methylene chloridej~.etlr.2nol, 10/1, is dissolved in 200 ml of 60b strength acetic acid and the solution is kep-t at 95-100C for 1.5 hours. After cooling, the solvent is distilled off. The residue is taken up twice more in a li-ttle water and in each case is evaporated to dr~ness. The resulting benzyl ester Gf benz~l-3-0-~D-l-rL-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carba~.oyl-ethyl~-2-deoxy--2-propionylamino-a-D-glucopyranoside is crystal-lised from methanol; melting point 189, [~]20 = +100 +1 (dimethylformamide9 c = 1.268).
Exam~le 2 A solution of 4 g of the benzyl ester of benzyl-2-acet-amido-3-0-~[L l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyl-methyl~ -2-deoxy-~-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 resi-due 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, urltil the reaction has ceased. The catalyst is filtered off and rinsed with a lit-tle water and the filtrate is evaporated to dryness. The resulting 2-acetamido-3-0-~L-l-(D-l-carbamoyl-3-carboxyl-propyl)-carbamoyl-ethyl]-carbamoyl-methyl~-2-deoxv-D-~, r ~ Q

glucose is dried over phosphorus pen~toxide ~mder a high vacuum;[a]20 -= -10 ~1 (wa-ter~ c = 0.93()).
The s-tar~ting ma-terial can be prepared as follows:
5.3 g of N-eth-yl-5-phenyl-isoxazolium 3'-sulphonate (Woodward's reagent K3 are added -to a solution of 9.5 g of benzyl-2 acetamido-4,6-0-benzylidene-3-0-carboxyme-thyl-2-deoxy-a-D-glucopyranosidein 400 ml of acetonitrile and 3 ml of triethyl-amine and the mixture is stirred at room ~temperature un-til a clear solution forms. 7.15 g of L-alanyl-D-glutamic acid 1--amide-r-benzyl ester-hydrochloride, 3 ml of triethylamine and 200 ml of acetonitrile are now added and the reaction mixJure 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-1-car-bamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyi-methyl~-2-deoxy-a-D-glucopyranoside, which has crystallised out, is filtered off, washed with half-saturated sodium bicarbonate solution and water and dried; ~a]D = +81 +1 (dimethylformamide, c = 0.816).
A solution of 8 g of this compound in 400 ml of 60%
strength acetic acid is kept at 80C for 1 hour. After cool-ing, the solu-tion is eva orated to dryness, a further t~ice 5G ml of wa-ter are added to the residue and in each case the mixture is evaporated to dryness. The resulting crystalline residue is stirred with a li-t-tle water and the crystals are filtered off and dried. This gives thebenzyl ester of benzyl-2-acetamido-3-0-~[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyl-methyl~-2-deoxy-a-D-glucopyranoside which has a melting point of 200-202, ~a]D = ~77 +l (dimethy~formamide, c = 0.599).

E;~m~le ~
~ solu-tion of 0.9 g of the benzyl ester of benzyl-2-acetamido-3-0-[L-l-(D-_ carbamoyl-3-carboxy-propyl)-carbamoyl-eth-yl~-carbamoyl-methyl-2-deoxy-~-0-stearoyl-~-D-glucopyranoside in 40 ml of methanol is hydrogena-ted under normal pressure and at room teMpera~tUre~ using 0.2 g of ~% strength palladium-on-charcoal as the catalyst, until 22.4 ml of hydrogen have been taken up. The catalyst is filtered off and rinsed wi-th me-tha-nol and the filtrate is evaporated to dryness. This gives benzyl-2-aceta~ido-3-0-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-e-thyl]-carbamoyl-methyl-2-deoxy-6-0-stearoyl-~-D-glucopyranoside, ~]20 = ~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-acetamido-3-0-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyl-methyl-2-deoxy-~-D-glucopyranoside in 30 ml of pyri-dine, 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-LL-l-(D-l-carbamoyl-3-carboxy-propyl)~carbamoyl-ethyl]-carbamoyl-,~d~

methyl-2-deoxy 6-0-stearoyl-a-D-glucopyranoside which has an [a]20 of +30 +1 (chloroform, c - 1.203).
Exam~le ~, Analogously to Example Z, benzyl-2-acetamido-4,6-0-benzylidene-3-0-(D-l-carbocy~propyl)-2-deoxy-a-D-glucopyranoside is subjected to a condensation reaction with L-alanyl-D-glutamic acid l-amide-r-benzyl ester-hydrochloride and the protective groups are split off. This gives 2-acetamido-3-0-~ [L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl~-carbamoyl-propyl~-2-deoxy-D-glucose.
The starting material can be prepared as follows:
10 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 60o ml of dimethylformamide and the mixture is stirred for 1.5 hours at 45C in a nitrogen atmosphere. After cooling to 0, 75 ml of ethyl D,L-a-bromobutyrate are added.
The reaction mixturc is stirred for 1 hour at room temperature and for 1 hour at 50C 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-4,6-0-benzylidene-3-0-(D-l-carboxy-propyl)-2-deoxy-a-D-glucopyranoside which has an ~]DO of ~113 +1 (chloroform, c = 0.5), a melting point of 15~C (from me-thyl2ne chloride/ether) and an Rf value of 0.21 and the ethyl ester of benzyl-2-ace~tamido-4,6-0-ben~Jli-dene-3-0-(L-l-carboxy propyl)-2-deoxy-Q-D-glucopyranoside which has an [a~D of +42 +1 (chloroform, c = 0.511), a mel-ting point of 240C (from ethyl aceta-te) and an Rf value of 0.04~
100 ml of lN sodium hydroxide solution are added -to a solution of 3$.1 g of the ethyl ester of benzyl-2-acetarnido-4,6-0-benzylidene-3-0-(D-l-carboxy-propyl)-2-deoxy-~-D-glucopyrano-side in 300 ml of methanol and the mixture is kept at 60 for 1 hour. The solution is then cooled, concen-trated to about 150 ml and diluted with 400 ml of ice water and 100 ml of a lN ice-cold 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-benzylidene-3-0-(D-l-carboxy-propyl)-2-deoxy-a-D-glucopyranoside which has a melting point of 210-213 and an [a]D of +110 -1 (dimethylformamide, c = 0.554).
Benzyl-~-acetamido-4,6-0-benzylidene-3-0-~L-l-carboxy-propyl)-2-deoxy-a-D-glucopyranoside which has a melting point of 285C and an [a]20 of +71 +1 (dimethylformamide, c = 0.589) is obtained in an analogous manner.

3.77 g of 2-phenyl-4,5-[3-0-(D-l-carboxyethyl)-5,6-0-isopropylidene-D-glucofurano~-~2-oxazoline in 60 ml of aceto-nitrile and 15 ml of dimeth-ylformamide are stirred together with 1.4 ml of triethylamine and 2.55 g of N-e-thyl-5-phenyl-isoxazolium 3'-sulphonate for 1.5 hours at 0, whereupon virtually everything goes into solution. 3.44 g of L-alanyl-D-glu-tamic acid 1-amide-~-benzyl ester-hydrochloride and a further 1.4 ml of triethylamine are then added and ~the mixture is s-tirred 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 7 firm syrup which crystallises on grinding with ether is obtained. Melting poin-t 96-99, Ca]D - -~13 (in chloroform).
The crystalline benzyl es-ter is hydrogenated, using 5,~
streng-th palladium-on-charco~l, 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 water with 10 ml of Dowex-50-H+ for 15 hours a-t room temperature. After filtering and freeze-drying, a colourless powder with a decomposition temperature of 140 is obtained. The 2-benzoylamino-3-0-~-D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-1-carbamoyl-ethyl]-carbamoyl-ethyl~-2-deoxy-~.~-D-glucose which is obtained contains varying amounts 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.
Exam~le 6

6.o g of 2-phenyl-4,5-~3-0-(D-l-carboxypropyl)-5,6-o-isopropylidene-D-glucofurano]-~2-oxazoline, 4.08 g of N-ethyl-5-phenyl-isoxazolium 3'-sulphonate and 2.25 ml of ~triethylamine are stirred in 100 ml of acetonitrile and 25 ml of dimethyl-formamide for 1 hour at 0-5C, whereupon the whole goes into solu-tion. 5.55 g of L-alanyl-D-glu~tamic acid l-amide-~-benzyl ester-hyclrochloride and a fur-ther 2.35 ml of triethylam:ine are then added and the mixture is stirred a-t room temperature for 48 hours. I-t is evaporated under an oil vacu~m and chromato-graphed on silica gel in a mic-ture of chloroform and ethanol (19/1). 9 g of the colourless syrup which is -thus o~tai~led are hydrogenated in dioxane using 5% strength palladium-on-charcoal, the catalys-t is filtered olf and the filtrate is con-centrated in vacuo and hydrolysed at room temperature in a ~i~ture of Lio ml of tetrahydrofurane and ~0 ml of water with 1.5 ml of trifluoroacetic acid. The water is -then evaporated to dryness 4 times in vacuo and the residue is dissolved in water and lyophilised. me r~slJlting 2-benzoylamino-3-0-~D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyl-propyl~-2-deoxy-~ D-glucose crystallises with 0.5 mol of water, melting point 114-152, [a~20 = ~17 (in me-thanol).
ExamPle 7 3.63 g of 2-phenyl-4,5-[3-0-carboxymethyl-5,6-o-iso-propylidene-D-glucofurano]-L~2-oxazoline, 3.43 g of L-alanyl-D-glutamic acid l-amide-~-benzyl ester-hydrochloride, 1.21 g of N-hydroxysuccinimide, 2.16 g of dicyclohexylcarbodiimide and 1.45 ml of triethylamine are dissolved in 40 ml of dime-thylformamide and the mixture is stirred for 24 hours at room temperature.
It is evaporated under an oil vacuum, the residue is ~taken up in dichloroe-thane and water, the dicyclohexylurea which has pre-cipitated ou-t is filtered off and the organic phase is twice extrac-ted by shaking wi-th ~;~ater and the aqueous phase is t~ice extrac-ted by shaking with dichloroethane~ Af~ter drying and evaporatirl~r the organic phases, a solid syrup is obtained and this is chroma~tographed on silica gel in a mix~ture of chloro-forrn/e-thanol (9/1). The resul-ting peptide ester, which crystallises on grinding ~rith e-ther, melts a~t 167-168, [~]20 = -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 fil-trates are evaporated and -the residue is recrystallised 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 Dowex-50-H+
for 15 hours at room temperature, the mix-ture 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-benzoyl-amino-3-0-~[L-l-(D-l-carbamoyl-3-carboxy-propyl)-l-carbamoyl-ethyl-carbamoyl-methyl~-2-deoxy-~ D-glucose which has a melting point of 175-177 (as the hydrate)~
The star-ting material can be prepared as follows, using a rnodification of the method described in Acta Chem. Scand. 18, 185 (1964):
100 g of 2-phenyl-4,5-[5,6-0-isopropylidene-D-gluco-furano]-~2-oxazo]ine are dissolved in 1 li-tre of acetonitrile, with the exclusion of moisture and carbon dioxide~ and 15.2 g of a 55,~ streng-th ~iaH/mineral oil dispersion are added in por-tions, whils-t s-tirring well, and the mixture is stirred for a _ ~4 _ further one hour at room tempera-ture. 42 ml of e~thyl chloro~
acetate are then added dropwise a~t 0 ancl after 1.5 hours a further 42 rnl are added dropwise. After 1.5 hours, a fur-ther 11.4 g OI -the NaH dispersion are added, ~the mix-ture is stirred fcr half an hour and a further 42 ml of the chloro-acetat~ are added dropwise at 0 ~ Af~ter a further 2 hours, the mix-ture is allowed -to warm to room temperature and is evapor-ated in vacuo - towards the end under an oil vacuum - to give a syrup. The lat-ter is taken up in ether and extracted 3 times by shaking with wa~ter, the ether phase is dried over sodium sul-phate and, after evaporating, 155 g of a brown oil are obtained.
This is dissolved in 150 ml of methanol and a solution of 30 g of potassium hydroxide in 150 ml of water is added, the mixture is 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 lN hydroGhloric acid, using a pH meter.
The crystalline 2-phenyl-495-[3-0-(carboxymethyl)-5,6-0-isopropylidene-D-glucofurano]-~ -oxazoline which has precipi-tated out is filtered off, washed with water and dried. 107 g, that is 93% of theory, with a melting point of 186-188, an []DO of -9 (CHC13, c = 0.9) and an [a]D of -23 (CHC13, c = 3) are obtained.
Exam~le 8 2.5 g of 2-benzamido-2-deoxy-3-0-[~-1-(D-l-carbamoyl-3-carboxy-propyl)-carbamoylethyl]-carbamoylmethyl-D-glucopyranose (Example 8) are dissolved in a mixture of 17 ml of absolute pyridine and 4 ml of butyric anhyclride. After 20 hours at room temperature and 2 hours at 50, water is added and the mix-ture is concentrated in vacuo. The residue is dissolved in chloroform, the chloroform solution is extracted by shaking with lN 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 polnt of 110-120 is thus obtained; this is a,~-1,4,6-tri-butyrGyl-2-benzamido-2-deoxy-3-0-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-car~amoylethyl~-carbamoylmethyl-D-glucop~ranose, Rf = 0.52 in CHC13:CH30H = 7:2 (silica gel thin layer, Merck).
Exam~le 9 2.3 g of 2-benzamido-2 deoxy-3-0-[L-1-(D-1 carbamoyl-3-carboxy-propyl)-carbamoylethyl]-carbamoylmethyl-D-glucopyranose are dissolved in 20 ml of pyridine and 5 ml of aeetic 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 CHC13:CH30H = 3:1 and a eolourless powder whieh has a melting point of 122-158 and an ~a~D of +48 (CH30H, c = 1.074) is obtained; this is a.~-1,4,6-triacetyl-2-benzamido-2-deoxy-3-0-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoylethyl~-carbamoylmethyl D-glueopyranose, Rf = 0.54, CHC13:CH30H = 3:2 (silica gel thin layer, Merck).
Exam~le 10 6.33 g of 2-phenyl-4,5-(3-0-carboxymethyl-5,6-o-iso-propylidene-D-glucofurano)-~2-oxazoline, 5.75 g of 2-ethoxy-N-carbethoxy-1,2-dihydroquinoline (EEDQ) and 9~3 ml of triethyl-~ 7~s amine are a-lded -to a solution of -the trifluoroacetate of dibenzyl L-alanyl-D-glutarna~te (obtained from 8.3 g of dibenzyl N-tert.-butoxycarbonyl-L-al.anyl-D-glutama-te and 5.1 ml. of trifluoro-acetic acid and 2.6 ml of dichloroe-thane by hydrolysing for 4 hours at LrO ) in 70 ml of dichloroe-thane. The mixture is allowed -to react at 40 for 15 hours and is diluted with chloro-form and twice extracted by shaking with water and the aqueous phases are extracted once by shaking with chloroformO After drying over sodium sulphate and evaporating the chloroform solu-ti.on, 19.9 g of an oil are obtained and this is purified over 400 g of Merck silica gel by elution with ether and then with chloroform:acetone, 17:3. ~liS gives pure 2-phenyl 4,5-[3-0-(1-L-~1-D,3-dibenzyloxy~carbonyl-propyl~ -carbamoylethyl)-carbamoylmethyl-5,6-0-isopropylidene-D-glucofurano]-~2-oxazoline which has a melting point of 113-116 and an [~]DO of 47 (CHC13, c = 1-54)-

7 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 of~`, 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 11 ml of Dowex-50 I,r x 4 ion exchanger in a mixture of 45 ml of tetrahydrofurane and 20 ml of water for 20 hours at 40. Af-ter filtering and clarifying with charcoal (Darco G 60), the solution is freeze-dried and colourless, amorphous 2-benzamido-2-deoxy-3-0~ L~ D,3-dicarboxy-propyl)-carbamoyl-ethyl]-carbamoylme-thyl-D-glucopyranose which has an optical rotation of [a]D = +25 (H20, c = 0.997), is obtained.

Ex,~ 1 Q 1 1 ~ nalogously to Example 10, 5.7 g of 2-phenyl-4,5[3-0-carbo~me~hyl-5,6-0-isopropylidene-D-glucofurano~-~ -o~azoline are subjected -to a condensa-tion reac~tion with 4.9 g of L-seryl-D-glutamic acid a-amide-r-tert.-butyl ester-hydrochloride wi-th the addition of 2.3 ml of ~triethylamine and 5.2 g of 2-etho~y-N-carboethoxy-1,2-dihydroquinoline in 45 ml of dichloroethane.
After 18 hours at 40 a crystalline product has formed. A
fur-ther 50 ml of dichloroethane are added and the mixture is cooled in ice and filtered and the crystals are washed with cold dichloroe-thane. The analytically pure, colourless crystals of 2-phenyl-4,5-[3-0-(1-L-~l-D-carbamoyl-3-tert.-butyloxy-carbonyl-propyl~-carbamoyl-2-hydro~xyethyl~-carbamoylmethyl-5,6-0-isopropylidene-D-glucofurano]-~2-oxazoline have a melting point of 187-188 and a [~]DO of +7 (CH30H, c = 1-125)-2 g of this compound are hydrolysed with a mixture of15 ml of methylene chloride and 5 ml of -trifluoroacetic acid for 20 hours at room temperature. The mixture is evaporated under an oil vacuum, the residue is ground with ether and 2-benzamido-2-deoxy-3-0-[1-L-(l-D-carbamoyl-3-carboxypropyl)-carbamoyl-2-hydroxyethyl]-carbamoylmethyl-D-glucopyranose is obtained as a beige powder with a melting point of 100-115, [~]DO = +23 (H20, c = 0 886), which crystallises with 2 mols of water and 1 mol of trifluoroacetic acid. f 3 3 (silica gel thin layer, Merck).
Exam~le 12 Analogously to Example 10, 5.25 g of 2-phenyl-4,5-[3-0--carboxymethyl-5,6-0-isopropylidene-D-glucofurano~ oxazoline and the trifluoroace-tic acid salt of L-alanyl-D-glutamic acid a-n-propylamide~-benzyl ester - obtained from 6.2 g of N-ter~t.-butoxy-carbonyl-L-alanyl-D-glutamic acid a-n-propylamide-~-benzyl ester and 4.2 ml of trifluoroace-tic acid in 2.5 ml of dichloro-ethane af-ter 6 hours a~t 40 - in 60 ml of dichloroethane are subjected -to a condensation reaction, with the addition of 7.75 ml of trie-thylamine and 4.8 g of 2-ethox-y-N-carboethoxy-1,2-dihydroquinoline. After 20 hours at 40, the mixture is dil-uted with 50 ml of chloroform and twice extracted by shaking with water and the water is twice extracted by shaking with chloro-form. After drying and concentrating the chloroform phase, 15 g of an 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 Rf of 0.35, CHC13:acetone = 7:3 (silica gel thin layer, Merck) are obtained.
This substance is hydrogenated in 80 ml of ~tetrahydro-furane 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 0.58, CHC13:CH30H = 3;1 (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 ~ternpera-ture and for 12 hours at 40 . ~fter filtering, clari-fying -the fil-trate with charcoal (Darco-G-60), fil~tering again and freeze--drying the filtrate 7 colourless, amorphous 2-benz-amido-2-deo~y-~-0-[1-L (1-D~T-r ~rGr~l-carbamoyl-3-carboxy-propyl) -carbamoylethyl]-carbamoyl-methyl-D-glucopyranose with a mel-ting point of 65-140 and an [a]20 of ~28 (water, c = 1.03) and an Rf of 0.48, CHC13:CH~OH = 1:1 ~silica gel thin layer, Merck' is obtained.
Exam~le 13 7.3 g of 2-phenyl-4,5-~3-0-carboxymethyl-5,6-G-isopropyl-idene-D-glucofurano]-Q2-oxazoline, 6.5 g of ~-amino-isobut~Jroyl-~-glutamic acid a-amide-1-tert.-butyl ester-hydrochloride and 2.9 g of isobutyl chloroformate are dissolved in 25 ml of dimethyl-formamide 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 -15 to -10 in the course of 30 minutes.
The 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 NaOH 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. ~`his is purified over Merck silica gel by elution with CHC13 C2H50H
= 19:1. 9.7 g of colourless, amorphous 2-phenyl-4,5-¦3-0-~1-methyl-l-(l-D-carbamoyl-3-tert.-butoxycarbonyl-propyl)-carbamoyl-e-thyl]-5,6-0-isopropylidene-3-glucofurano~-Q2-oxazoline which has an op-tica:L rota~tion: ~a]D = +6 (CHC13, c = 1.027), a melting poin-t of 75-89 and an R~ of 0.35, CHCl~:C2H50H - 9:1 (silica gel thin layer, Merck) are ob-tained.

8.3 g of the above compound are allowed to stand in a mixture of 20 ml of trifluoroacetic acid9 60 ml of methylene chloride and Z ml of water for 15 hour~ at room temperature.
The mix-ture is then evaporated in vacuo and the residue is ground with ether. 1~1e resultlng 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 filtra-te, colourless, amorphous 2-benzamido-2-deoxy-3-0-[1-methyl-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoylethyl]-carbamoylmethyl-D-glucopyranose with a melting point of 110-120, an [a]D of f31 (H20, c = 0.88) and an Rf of 0.52, acetone:
ethanol = 1:1 (silica gel thin layer, Merck), which crystallises with o~6 mol of trifluoroacetic acid and 1.7 mols of water~is obtained.
Exam~le 14 Analogously to Example 10, 2-benzamido-2-deoxy-3-0-carboxymethyl-a-ethyl-D-glucopyranoside and the trifluoroacetic acid salt of L-alanyl-D-glutamic acid a-amide-~-tert.-butyl es-ter with 2-ethoxy-N-carboethoxy-1,2-dihydroquinoline give the corresponding glycopeptide which has an [a~Dof -23 (CH30H, c =
1.107) and an Rf of o.L~7 (CH2C12:C2H50H = 8:2~ and, after hydro-genation ln tetrahydrofurane/water using 5~ strength Pd/C, the glyco-peptide gives 2-benzamido-2-deoxy-3-0-[L-lr-(D-l-carbamoyl-3-carboxy--propyl)-carbamoylethyl~-carbamoylmethyl-~-ethyl-D-~ a ~ ~ ~ 7 ~

glucopyranoside which has a melting psint of 215-217, an L~1~
of -22 (CH30H, c = 0.97) and an Rf of 0.36 in CHC13:CH30H = 1:1 (silica gel thin layer, Merck).
2-Benzamido-2-deoxy-3-0-carboxymethyl-~-ethyl-D-gluco-pyranoside which is used as the s-tar-ting material is obtained as follows:
2-Phenyl-4,5-~3=-0-carboxymethyl-5,6-0-isopropylidene-D-glucofurano]-~ -oxazoline is dissolved in 0.1 N HCl/C2H50H
and the solu-tion is left to stand for 6 hours at room tempera-tureO I-t is neutralised with sodium e~thylate in ethanol and evaporated to dryness and the residue is taken up in acetone.
m e solution is filtered through a layer of Merck silica gel, the eluate is evaporated to dryness and the residue is extracted twice with e-ther at room tempera-ture. After recrystallisa-tion from ethyl acetate~ 2-benzamido-2-deo~y-3-0-carboethoxymethyl-~-ethyl-D-glucopyranoside which has a melting point of 185-188 and an [a]D = -35 (CH30H, c = 1.121) is obtained.

9.4 g of this ester are saponified with a solu~tion of 1.7 g of po-tassium hydroxide in 250 ml of ethanol and 25 ml of water for 2 hours at room temperature. The pH value is then adjusted to 3.5 with 1 N hydrochloric acid and the mixture is concentrated in vacuo.
The residue is first ground with e-ther and is then ground wi-th 3 times 20 ml of ice water and the mixture is filtered.
Crystals of 2-benzamido-2-deoxy-3-0-carboxymethyl-~-e~thyl-D-glucopyranoside with a mel-ting point of 205-210 and an [~]DO
of -40 (CH30H, c = 1.04) are thus obtained.

2-Benzamido-2-deoxy-3 0-[~D-l-carbamoyl~3-carboxy-propyl)-carbamoylme-thyl]-carbamoy:Lmethyl-D-glllcose is obtained from 3 g of 2-phenyl-4,5-[3-0-~(D-l-carbamoyl-3-carboxy-propyl)-carbamoylmethyl3-carbamoylmethyl-5,5-0-isopropylidene-D-gluco-furano~-~2--oxazoline by hydrolysis wi-th ].5 ml of trifluoro-acetic acid in a mixture of 15 ml of dimethoxyethane and 15 ml of water at 40 for 3 hours. m e reaction mixture is concen-trated to dryness in vacuo and the residue is again extracted with ether. The residual powder is dissolved in water and the solution is trea-ted with Darco-G-60 charcoal and filtered and the filtrate is freeze-dried. This gives a colourless, amorphous substance with a melting point of 115-155 and an [a]D of ~34 (water, c = 0.81), Rf = 0.28 CHC13:CH30H = 1:1 (silica gel thin layer, Merck).
m e starting material for this reac-tion is obtained as ~
follows:
8.0 g of N-tert.-butoxycarbonyl-glycyl-D-glutamic acid ~-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-N-carboethox~-1,2-dihydroquinoline (EEDQ) and 8.1 g of 2-phenyl-4,5-[3-0-carboxymethyl-5,6-0-isopropylidene-D-glucofuranol~-~2-oxazoline and 20 ml of dimethylformamide are addedto the solution, whilst cooling. After 20 hours at 40, the mixture is con-centrated under an oil vacuum and the residue is partitioned , ~_ f 3 ~ ' ? ,~ Q

between metnylerle chloride and wa-t;er. After drying and cGncentra-ting the me-thylene chloride phase5, a solid resldue is o~,tained and this is ex~tracted twlce with ether and recrystal-lised from toluene. Yield 8.2'j g; mel-ting poin-t 1579 [a~20 = +10 (CHC13, c = 1.48), Rf = 0.35 (CHC13:e-thanol = 9;1) (silica gel thin layer, Merck).
The benzyl es~ter which 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 fil tering off the catalyst and evaporating the filtrate, the substance is chromatographed on 250 g of Merck silica gel in CHC13:CH30H = 4:1- 5.8 g of colourless, amorphous 2-phenyl-4,5-~3~0-`(D-l-carbamoyl-3-carboxy-propyl) carbamoylmethyl~ -carbamoylme-thyl-5,6-0-isopropylidene-D-glucofurano]-~2-oxazoline which has an Rf of 0.43, CHC13~CH30H = 3:2 (silica gel thin layer, Merck) are obtalned.
Exam~le_16 Analogously to Example 10, 9.5 g of 2-phenyl-4,5-C3-0-carboxymethyl-5,6-0-isopropylidene-D-glucofurano~-~2-oxazoline are subjected to a condensation reaction with 6.25 g of L-alanyl-D-glutamic acid ~,~-diamide-hydrochloride with the addi-tion of 3.4 ml of triethylamine and 7.95 g of 2-ethoxy-N-carboethoxy-1,2-dihydroquinoline (EEDQ) in a mixture of 50 ml of dichloroethane and 150 ml of dimethylformamide. The mix-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 e-tner an~i ~then twice ~ith ice -~Jater. ~f-ter drying, -the pro-duc-c can be recrys-tallised from dlchloroethane~ This gives colourless crystals which have a melting point of 170-184;
[~]D0 _ +3 (DMS0, c = 1.43), Rf -- 0~64 CHC13:CH30H = 3:1 (silica gel thin layer, Merck).
6.1 g of this compound are hydrolysed with 13.5 ml of Dowex 50 ion exchanger in a mixture of 60 m] of dimethoxyethane and 60 ml of water for 15 hours at room -temperature. ~f-ter filtering and concentrating the filtrate, the residue is taken up in water, the mix~ure 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-1,3-dicarbamoyl-propyl)-carbamoylethyl]-carbamoylmethyl-D-gluco-pyranose whic'n has a melting point of 82-143; [~]DO = +24 (H20, c = 0.98) 9 Rf = 0.45 CHC13:GH30H = 1:1 (silica gel thin layer, Merck). m e substance crystallises with 1.23 mols of~
water of crystallisation.
Exam~le 17 Analogously to Example 16, ethyl-2-benzamido-2-deoxy-3 0-carboxymethyl-~-D-glucopyranoside and L-alanyl-D-glutamic acid ~,~-bis-methylamide-hydrochloride give ~-ethyl-2-benzamido-2-deoxy-3-0-[L-l-(D-1,3-bis-N-methyl-carbamoyl-propyl)-carbamoyl-ethyl]-carbamoylmethyl-D-glucopyranoside which has a melting point of 233, an [~]DO of -20 (CH30H, c = 0.937) and an Rf of 0.38 in CHC13:ethanol, 7:3 (silica gel 9 thin layer plates, Merck).

.

:~ ~d~
~,~cam~le 1 ~
Analogollsly to Example 16, 2-benzamido-2-deoxy-3-0-carboxyme~thyl-D-glu50pyranose gives 2-benzamido-2 deoxy-3-0-[L-l-(D-1,3-bis-N-m2thylcarbamoyl-propyl)-carbamoylethyl]-carbamoylmethyl-D-glucopyranose which has a melting point of 125-132, and [~]20 of and an Rf of 0.25 in CHC13:ethanol = 7:3 (silica gel -thin layer pla-tes 9 Merck).
Example 19 Analogously to Example 16, 2-benzamido-2-deoxy-3-0-carboxyrne~thyl-D-glucopyranose and the hydrochloride of dimethyl L-alanyl-D~glutamate gi~e 2-benzamido-2-deoxy-3-0-[L-l-(D-1,3-bis-methoxycarbonyl-propyl)-carbamoyl-ethyl]-carbamoylmethyl-D-glucopyranose in the form of the hydrate with a melting point of 80-90, an [~]DO of ~25 (CH30H, c = 1.017) and an Rf value of 0.23 in CHC13:e~thanol = 9:1 (silica gel thin layer plates, Merck).
Exam~le 20 Analogously to Example 16, ethyl-2-benzamido-2-deoxy-3-0-carboxymethyl-~-D-glucopyranoside gives ethyl-2-benzamido-2-deoxy-3-0-[L-l-(D-1,3-bis-methoxy-carbonyl-propyl)-carbamoylethyl]-carbamoylmethyl-~-D-glucopyranoside which has a melting point of 127-135, an [~]DO of -17 (CH30H, c = 10024) and an Rf of 0.26 in ethJl acetate:acetone = 2:1 (silica gel thin layer plates, Merck).
Exam~le 21 2-Benzamido-2-deoxy-3-0-~L-l-(D-l-N-benzyl-carbamoyl-3-carboxy-propyl)-carbamoyle-thyl]-carbamoylmethyl-D-glucopyranose is ob-tained analogously to Example 12.

In this case, the ~-carbo~yl group of the glutamic acid is liberated from a ter-t.-butyl es-ter by saponification ~ith Do~i~ex-50-H' in tetrah~drofurane/wa-ter.
Exam~le ~2 2-Benzamido-2-deoxy-3-0-[L-l-(D-l-N~carbamoylmethyl-carbariloyl-3-carboxypropyl)-carbamoyle-thyl]-carbamoylmethyl-D-glucopyranose is obtained analogously to Example 12. The star-ting ma-terial for the peptide part is N-tert.-bu-to,~ycarbonyl-L-alanyl-D-glutamic acid ~-benzyl es~ter-~-glycine-amide.
Exam~le 23 Analogously to Example 12, the hydrochloride of L-~-amino-valeroyl-D-glutamic acid ~-amide-r tert.-butyl ester gives 2-benzamido-2-deoxy-3-0- CL~1_ (D-l-carbamoyl-3-carboxy-propyl)-carbamoylbutyl]-carbamoylmethyl-D-glucopyranose.
Exam~le 24 2-Benzamido-2-deoxy-3-0-[L-l-(D~l-carbamoyl-3-carboxy-propyl)-carbamoyl-propyl]-carbamoylmethyl-D-glucopyranose is obtained analogously to Example 12.
Exam~le 25 2-Benzamido-2-deoxy-3-0-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-2-methylpropyl3-1-~bamoylme~hyl-D-gluc~nose is obtained analogously to Example 12.
Example 26 A solution of 10.7 g OI the benzyl ester of benzyl-2-acetamido-4,6-0-benzylidene-3-0-[(D-l-carbamoyl-3-carbox-y-propyl)-carbamoylmethyl-carbamoylmethyl]-2-deoxy-~-D-gluco-pyranoside in 200 ml of glacial acetic acid and lO0 ml of water J ~

is hydrogenated under normal pressure and a~t 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 fil-trate is evaporated. The resiclue is taken up in water and-the mixture is filtered throu~h lOG ml of ~nberlite IR 120 (H+ form) ion exchanger and the filtrate is freeze-dried. 2-~cetamido-3-0-~(D-l-carbamoyl-3-Garboxy-propyl)-carbamoylmet~lyl~carbamoyl-methyl]-2-deoxy-D-g]ucose is crystallised from methanol/ethyl acetate and dried under a high vacuum. The product, which contains 1/4 mol of ethyl acetate, displays an optical rota-tion [~]D0 = +27 +1 (water, c = 0.944).
The starting material used can be prepared as follows:
8 g of the benzyl ester of N-t-butoxycarbonyl-glycyl-D-isoglu-tamine are dissolved, at room temperature and with the exclusion of moisture, in a mixture of 18 ml of 1,2-dichloroethane and 8.4 ml of trifluoroacetic acid and 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-acetamido-4,6-0-benzylidene-3-0-carboxy-methyl-2-deoxy-~-D-glucopyranoside in 100 ml of tetrahydrofurane and 2.52 ml of triethylamine is added. After adding 5.05 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ), the mixture is stirred for 24 hours at room tempera-ture. 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-benzyl-idene-3-0-~(D-l-carbamoyl-3-carboxypropyl)-carbamoylmethyl-carba~oyl~,e~thyl] 2-deoxy-~-D-glucopyranoside has an optical rotation ~]20 = +66 ~1 (N,N-dimethylformaMide, c = 1.308).
F~am~le Z7 A solution of 4.5 g of the benzyl ester of methyl-2-acetamido-3-0-¦[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoyl-ethyl]-carbamoylmethyl}-2-deoxy-a-D-glucopyranoside in 125 ml of 50a~ strength aqueous methanol is hydrogena-ted under normal pres-sure and at room tempera-ture, using 1.0 g of 5% streng-th palladium-on-charcoal, until 178 ml of hydrogen has been taken up. I'he catalyst is filtered off and the filtrate is evapor-ated under a waterpurnp vacuum. The residue is dissolved in 50 ml of dis~tilled water and freeze-dried. This gives methyl-2-acelamido-3-0-~[L-l-(D-l-carbamoy].-3-carboxypropyl)-carbamoyl-ethy]]-carbamoylmethyl~-2-deoxy-~-D-glucopyranoside which has an [~D0 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-L-alanyl-D-isoglutamine are dissolved, at room temperature and with the exclusion of moisture, in a mixture of 8.1 ml of 1,2-dichloroethane and 8.1 ml of trifluoroacetic acid and the solu-tion is left to stand for 16 hours. The reaction mixture is diluted with 200 ml of tetrahydrofurane and, with external cool-ing, neutralised with triethylamine and a solution of 7.62 g of methyl-2~acetamido-4,6-o-benzylidene-3-0-carboxymethyl-2-deoxy-~-D-glucopyranoside and 2.77 ml of triethylamine in 100 ml of tetrahydrofurane is added. After adding 5.0 g of 2-ethoxy-N-e-thoxycarbonyl-1,2-dihydroquinoline (EEDQ), ~the reaction rnixture is warmed -to 40 , s-tirred at this temperature for 30 hours and left to stand for a further 24 hours at room tempera-ture. The product whlch has precipitated ou-t7 tha~t is to say the benzyl ester of methyl-2-acetamido-4,6-0-benzylidene-3-0-~L-1-(D-l-carbamoyl-3-carbo~propyl)-carbamoyl-ethyl]-carbamoyl-methyl~-2-deoxy-a-D-glucopyranoslde 7 iS filtered off, washed with tetrahydrofurane and ether and dried9 ~a]20 = +58 ~1 (N,N-dime-thylformamide, c = 1.125).
A solution of 10.5 g of the benzyl ester of methyl-2-acetamido-4,6-0-benzylidene-3-0-~[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyl-methyl~ -2-deoxy-a-D-gluco-pyranoside in 320 ml of glacial acetic acid is diluted with 200 ml of water, whilst stirring, and -the whole is stirred at 50-55C
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 es~ter of methyl-2-acetamido-3-0-~[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyl-methyl~-2-deoxy-a-D-glucopyranoside which has an [a]20 of +64 -1 (N,N-dimethylformamide, c = 1.26~).
Exam~le 28 A solution of 3.0 g of the benzyl ester of me-thyl-2-acetamido-3-0-~[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoyl-ethyll-carbamoyl-me-thylJ-2-deoxy-6-0-stearoyl-a-D-glucopyranoside in 100 rnl of ethanol and 100 ml of tetrahydrofurane is hydrogen-ated at room temperature and under normal pressure using 0.6 g of 5% strength palladium-on-charcoal, the catalyst is filtered ~ 3~

off and the fil-trate is evaporatecL to dryness under a water pump vacuum. The crystalline residue is methyl-2-acetamido-3-0-~[L-l-(D-1-carbamoyl-3-carboxypropyl~-carbamoyl-e-thyl]-carbamoyl-methyl~-2-deoxy-6-0-stearoyl-a-D-glucopyranoside which has an [a~20 of +50 ~1 (N,N-dime-thylformamide, c = 0.921).
The 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 3 hours to a solution of 3.98 g of the benzyl ester of methyl-2-acetamido-3-0-~[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyl-methyl~-2-deoxy-a-D-glucopyranosidein40 ml of absolute pyridine~ whilst s-tirring and with the exclusion of moisture7 and the mixture is lef~t to stand for 18 hours at room temperature. The reaction mixture is diluted with chloro-form, washed with water, ice-cold 2 N hydrochloric acid and water, dried over magnesium sulphate and evaporated to dryness. The product, that is to say the benzyl ester of methyl-2-acetamido-3-0-~L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoyl-ethyl]-carbamoyl-methyl)-2-deoxy-6-0-stearoyl-a-D-glucopyranoside7 is crystallised from ethanol/ether; [a]D = +22 +1 (chloroform, c = 1.030).
Exam~le 29 A solution of 6.8 g of benzyl-2-acetamido-3-0-~[~-1-(D-1,3-dicarbamoylpropyl)-carbamoylethyl]-carbamoylmethyl~-2-deoxy-a-D-glucopyranoside in 200 ml of 50% strength aqueous methanol is hydrogenated under normal pressure at room tempera-ture for ~0 hours using 5,~ strength palladium-on charcoal.

.~ A~ J ~

The catalys-t is fil-tered off and the filtrate is evaporatedO
The re.cidue is taken up in 50 ml of water and lyophilised.
This gives 2-acetamido-3-0-~[L-l-~D-1~3-dicarbamoylpropyl)-carbamoylethyll-carbamoylmethyl~-2-deoxy-D-glucose, which con-tains 1~24 mols of water, in the form of a white powder with an [~l20 O~ ~7 +1 (water~ c = 0.514).
The starting material can be prepared as follows:
5.0 g of L-alanyl-D-glutamic acid diamide-hydrochloride and 5.1 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) are added to a solution of 9.1 g of benzyl-2-acetamido-4,6-0-benzylidene-3-0-carboxymethyl-2-deoxy-a-D-glucopyranoside in 100 ml of N,N-dimethylformamide and 2 77 ml of triethyl-amine 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 water and the product is dried and recrystallised from chloroform/methanol; [a]23 =
+83 -1 (N,N-dimethylformamide, c = 0.531~.
A solution of 4 g of benzyl-2-acetamido-4,6-0-benzylidene-3-0-~L-l-(D-1,3-dicarbamoylpropyl)-carbamoylethyl~-carbamoyl-methyl3-2-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 resul-ting benzyl-2-acetamido-3-0-~L-l-(D-1,3-dicarb3moylpropyl~calbamoylethyl]-carbamoyl-methyl~-2-deoxy-~-D-glucopyranoside is recrystallised from methanol; melting point 223-225.

~ ~ ~ Z ~

Exam~le 30 A solution of 5.7 g of ~the benzyl ester of benzyl-2-ace-tamido-3-0-~D-l-[(D-1-carbamoy]~3-carbox-ypropyl)-carbamoyl-methyl]-carbamoylpropyl~-2-deoxy-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. The catalyst is filtered off and the filtrate is evaporated. The resldual 2-acetamido-3~0-fD-l-~(D-l-carbamoyl-3-carboxypropyl)-carbamoylmethyl]-carbamoylpropyl3-2-deoxy-D-glucose is taken up ln 50 ml of water and freeze-dried; [a]D
= +46 T1 (water, c = 0.630).
The starting material used can be prepared as follows:
5~1 g of the benzyl ester of N-t-butoxycarbonyl-glycyl-D-iso-glutamine are dissolved, at room temperature and with the exclu-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 tetrahydrofurane and, with external cooling, neutralised with triethylamine and a solution of 6.3 g of benzyl-2-acetamido-4,6-0-benzylidene-3-0-(D-l-carboxypropyl)-2-deoxy-a-D-glucopyranoside and 1.8 ml of triethylamine in 100 ml of tetrahydrofurane and 3.2 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) is added. After 24 hours, the mixture is evaporated to dry-ness 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 bicar-bonate and wa-ter and evaporated. The residual benzyl ester of benzyl-2-acetar.~ido-Li,6-O~benzy:Lidene-3-0-~D-l-[(D-l-carbamoyl-3-carboxypropyl)-carbamoylmethyi]--carbamoylpropyl} -2-deoxy-~-D-glucopyranoside is crystallised from te-trahydrofurane/ether~
~]20 +76 +1 (N,N-dimethylformamide, c = 00457).
Mild acid hydrolysis of this product in 60,~ strength aqueou~ acetic acid leads to the benzyl ester of benzyl-2-acetamido-3-0--~D-l-[(D-l-carbamoyl-3-carboxyprop~,rl)-carbamoyl-methyl]-carbamoylpropyl3 -2-deoxy-a-D-glucopyranoside) which crystallises from methanol/ether; melting poin-t 180-185, ~OL]D = -~87 +1 (methanol, c = 1.035).
Exam~le 31 4 g of the benzyl ester of N-t-butoxycarbonyl-L-alanyl-D-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-c~-D-glucopyranoside and 1.38 ml of trie~thylamine in 100 ml of tetrahydrofurane is added.
After adding 2.6 g of 2-ethoxy-N-e~thoxycarbonyl-19 2-dihydro-quinoline (EEDQ), the mixture is left to stand for 24 hours at room temperature and is evaporated -to dryness. The residue is dissolved in chloroform/me-thanol, 9/1, and this solution is washed with water, ice-cold 2 N hydrochloric acid, water, a sa-turated solution of sodium bicarbonate and water and the sol-vent is evaporated. The benzyl ester of benzyl-2-acetamido-t~

3-0-~l_-1-(D-]-carbamoyl-3-carboxypropyl~-carbamoyl-ethyl]-carbamoylmet.h~l~-2-deoxy-~-D-glucc)pyranoside, which is thus Gb-tained, is recrys~tallised from ethanol; melting point 208-212~ ~c~]20 = +77 +1 (N,N-dimethylformamide, c = 0.546).
Af-ter spli-tting off the t~Jo benzyl radicals by hydro-genolysis, as described in ExampLe 2, 2-ace-tamldo-~-0-~L-l-(D-l-carbamoyl-3-carbocypropyl)-carbamoylethyl]-carbamoylmethyl3-2-deoxy-D-glucose is ohtained.
Exam~le 32 3.8 g of the dimethyl ester of benzyl-2-acetamido-3-0-~[L l-(D-1,3-dicarboxypropyl)-carbamoylethyl]-carbamoylmethylJ-2-deoxy-~-D-glucopyranoside are dissolved in 100 ml of methanol and hydrogenated under normal pressure and at room temperature in the presence of 5% strength palladium-on-charcoal After the absorption of hydrogen has ceased, the catalys~t is filtered off and the filtrate is e-~aporated to dryness. The residue is taken up in 70 ml of water and freeze-dried. The resul-ting foam is the dimethyl ester of 2-acetamido-3-0-~[L-l-(D-1,3-dicarboxypropyl)-carbamoylethyl]-carbamoylmethyl~-2-deoxy-D-glucose which has an [a]D of +23 +1 (water, 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 6095 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) are added to a solution of 12.9 g of benzyl-2-acetamido-4,6-0-benzylidene-3-0-carboxymethyl-2-deoxy-~-D-glucopyranoside and 4.0 ml of triethylamine in 100 ml of N,N-dimethylformamide and the mic-ture is lef-t to stand at room temperature for 20 ~ t~

hours. The solvent is then evaporated, the residue is taken up ln chloroform and this solution is washed with water, ice-cold 2 N hydrochloric acid, wa-ter, a sa-turated solu-tion of sodium bicarbonate and wa~ter. After drying over magnesium sulphate~ the solution is evaporated -to dryness. The residue is extrac~ted with warm e~thanol ancl the dimethyl ester of benzy]-2-acetamido-4,6-0-benzylidene-3-0-~[L-l-(D-1,3-dicarbo~ypropyl)-carbamoylethyl]-carbamoylmethyl~-2 deoxy-a-D-glucopyranoside, which has no-t dissolved, is filtered off and dried; [a]D =
+22 -1 (chloroform, c = 1.160).
15~6 g of the dimethyl ester of benzyl-2 acetamido-4,6-0-benzylidene-3-0-~[L-l~(D-1,3-dicarboxypropyl)-carbamoyl-ethyl]-carbamoylme-thyl3-2-deoxy-a-D-glucopyranoside are dis-solved in 420 ml of glacial acetic acid and 280 ml of water and the solution is warmed to 10C. After stirring for 4 hours at this temperature, the reaction mixture is cooled and evapor~
ated to dryness. A further three times 100 ml of wa~ter are added to the residue and the mixture is evaporated to dryness each time. The residue is ~then taken up in chloroform and this solution is washed with water, dried over magnesium sul-phate and evaporated to dryness. This gives the dimethyl ester of benzyl-2-acetamido-3-0-¦[L-l-(D-1,3-dicarboxypropyl)-carbamoyle~thyl]-carbamoylmethyl}-2-deoxy-a-D-glucopyranoside as a yellowish resin with an [a]D of +31 +1 (chloroform, c = 1.070).
Exam~le 33 A solution of 6.1 g of the benzyl ester of benzyl-3-0-~L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoyle~thyl]-carbamoyl-methyl~-2-deoxy-2-propionamido-~-D-glucopyranoside in 200 ml of 2/1 tetrahydrofurane/water is hydrogenated under normal pres-sure 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 ca-talys-t is filtered off and the fil-trate is evaporated. 150 ml of water are added to the resi-due and the mixture is hydrogenated in the presence of 5~ strength palladium-on-charcoal until no further hydrogen is taken up.
The catalys-t is filtered off and the filtrate is lyophilised.
This gives 3-0-~[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoyl-ethyl~-carbamoylmethyl}-2-deoxy-2-propionamido-D-glucose.

The startlng material used can be prepared as follows:
0.3 ml OI methanesulphonic acid is added to a solution of 90 g of benzyl-2-acetamido-2-deoxy-~-D-glucopyranoside in 900 ml of N,N-dimethylformamide~ whilst stirring, with the exclu-sion of moisture and whilst cooling with ice water. A solution of 60 ml of isopropenyl methyl ether in 2~0 ml of N,N-dimethyl-formamide 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 benzyl-2~acetamido-2--deoxy-L~,6-0-isopropylidene-~-D-glucopyranoside is crystallised from ether; mel-ting point 136-137, [a~D = +103 ~ ~''3~

+1 (chloroform, c = 1.125) 52.5 g of benzyl-2-acetamido-2-deoxy-4,6-0-isopropyli-dene-~-~-glucopyranoside are dissolved in a solution of 225 g of potassium hydroxide in 750 ~1 of ethanol and 40 ml of dis-tilled water and the solution is boiled under reflux for 42 hours. After cooling, the reac-tion mixture is concentrated to half its volume and poured onto ice. The mixture is extracted with chloroforrn and the organic phase is washed .~ith water, dried over magnesium sulphate and evaporated -to dryness The residue, that is to say benzyl-2-amino-2-deoxy-4,6-0-iso-propylidene-a-D-glucopyranoside, is crystallised from ether;
melting point 145-146CJ [~20 = +117 +1 (chloroform, 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 benzyl-2-amino-2-deoxy-4,6-0-isopropylidene-~-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 sul-phate and evaporated to dryness. The product, that is to say benzyl-2-deoxy-4,6-0-isopropylidene-2-propionamido-~-D-gluco~
pyranoside~ is crystallised from ethyl acetate/petroleum ether;
melting point 121-122, [a]D = +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-O~isopropylidene-2-propionamido-a-D-o~lucopyranoside in 90 ml of acetoni-trile and the mixture is stirred for 2 hours at 40 . It is then cooled to 5 to -10C and 4.2 ml of ethyl bromoacetate are added.
After a further 20 minu-tes, 10 ml of ethanol are added and the reaction mixture is neutralised with glacial acetic acid and evaporated to dryness. The residue is parti-tioned between e-ther and water and the ether solution is washed wi-th water, dried over magneslum sulphate and evaporated. The produc-t, that is to say the ethyl ester of benzyl-3-0-carbo~Jmethyl-2-deoxy-4,6-0-isopropylidene-2-propionamido-a-D-glucopyranoside, is crystallised from ether/petroleum e-ther; melting point 94-95~ Ca]20 = +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-3-0-carboxymethyl-2-deoxy-4,6-0-isopropylidene-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 reac-tion with 15 mmols of L-alanine-D-isoglutamine benzyl ester-trifluoroacetate in the presence of 3.72 g of 2-ethoxy-N~ethoxy-carbonyl-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 bicar-bonate and water, dried over magnesium sulphate and evaporated to dryness. The produc-t is crystallised from dilute ethanol, melting poin-t 177-~0, [~]20 = +7:l ~1 (chloroform, c = 1.047).
15 ml of 1 N hydrochloric acld are added -to a solution of ~.1 g of the benzyl ester of benz-yl-3-0-~rL-l-(D-l-carbamo~l-3-carboxypropyl)-carbamoylethyl~-carbamoylmethyl}-2-deo~y-4,6-0-isopropylidene-2-propionamido--~-D-glucopyranosi.de in 150 ml of methanol and the mixture is l.ef-t to s~tand for one hour at room tempera-ture. 15 ml of 1 N sodium hydroxide solution are now added and the mix-ture is evaporated -to dryness. The resulting benzyl ester of benzyl-3-0-~[L-l-(D-l-carbamo~l.-3-carbo~ypropyl)-carbamoyl-ethyl~-carbamoylmethyl3-2-deoxy-2-propionamido-a-D-glucopyranoside is crystallised from me-thanol/
water and dried; melting point 20~-210, [~]DO = ~75 +1 (N,N-dimethylformamide, c = 1.120)~
Exam~le 34 3-0-~[L-l-(D-1 Carbamoyl-3-carboxypropyl)~carbamoylethyll-carbamoylmethyl~-2-deoxy-2-caprinoyl-amido-D-glucose is obtained as a white foam , by catalytic hydro-genation of the benzyl ester of benzyl-3-0-~[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoylethyl~-carbamoylmethyl~-2-deoxy-2-caprinoylamldo-~-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-D-glucopyranoside are reacted with 16O7 g of caprinoyl chloride and the reac~tion mixture is worked up. This gives benzyl-2-deoxy-4,6-0-isopropylidene-2-caprinoylamido-a-D-glucopyranoside which has an [a~20 of +81 ~1 (chloroform, c = 1.109).
o . L~ g of pract. sodium hyclride is added to a solution of 3.8 g of benzyl-2-deoxy-4,6-0-isopropylidene-2-caprinoyl-amido-a-D-glucopyranoside in 40 ml of ace-tonitrile and the mixture is stirred for 2 hours at 40O This mixture is then cooled -to -10, 1.4 ml o~ ethyl bromoacetate are added and the mixture is stirred for a fur-ther hour at 0. ~en the mixture is worked up as described in Example 33, this gives the ethyl ester of benzyl-~-O-carboxymethyl-4,6-0-isopropyli~
dene-2-caprinoylamido-a-D-glucopyranoside as a yellow-brown oil with an [a]20 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 L-alanine-D-isoglutamine benzyl ester-trifluoroacetate in the pre-sence of EEDQ and the isopropylidene-ketal is split by mild acid hydrolysis. In this way the benzyl ester of benzyl-3-0-~[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoylethyl]-carbamoylmethyl~-2-deoxy-2-caprinoylamido-a-D-glucopyranoside is obtained as a yellowish resin.
ExamPle 35 A 5% strength solution of benzyl-2-acetamido-2-deoxy-3-0-~[L-l-(D-1,3-bis-methylcarbamoylpropyl)-carbamoylethyl]-carbamoylmethyl3-a-D-glucopyranoside in 1/1 distilled water/
methanol is hydrogenated using 5~0 strength palladium-on-charcoal, -the catalyst is filtered off and the filtrate is evaporated. The product~ that is to say 2 acetamido-2-deoxy-3-0-~[L-l-(D-1,3-bis-methylcarbamoylpropyl)-carbamoyl-ethyl]-carbamoylmethyl~-D-glucose is freeze-dried; Ca~D =

The starting material can be prepared as ~ollows:
5.5 g of L~alanyl-D-glutamic acid bis-me-thylamide-hydrochloride and 5.1 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline are added to a solution of 9.1 g of benzyl-2-acetamido-4,6-0-benzylidene-3-0-carbo~Jmethyl-2-deoxy-a-D-glucopyranoside in 100 ml of N,N-dime-thylformamide and 2.77 ml of triethyl~mine and~he ~ ture~sleft to stand for 48 hours at room temperature. The solvent is now distilled ofî, l~ater is added to the oily residue and the insoluble matter is filtered off, washed with water and dried. This product is stirred twice mor~ with ether, filtered off and dried and is benzyl-2-acetamido-4,6-0-benzylidene-2-deoxy-3-0-~[L-l-(D-1~3-bis-methylcarbamoylpropyl)-carbamoylethyl]-carbamoyl-methyl3-a-D-glucopyranoside.
By means of mild acid hydrolysis, as described in Example 29, with 60,6 strength acetic acid, the benzylidene group is split of and the product, that is to say benzyl-2-acetamido-2-deoxy-3-0-~L-l-(D-1,3-bis-methylcarbamoylpropyl)-carbamoylethyl]-carbamoylme-thyl~-a-D-glucopyranoside is iso-lated.
Exam~le 36 A 5,6 strength aqueous solu-tion of benzyl-2-acetamido-3-0-~[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoyl-2-hydroxy-ethyl]-carbamoylmethyl~-2-deoxy-a-D-glucopyranoside is hydro-genated in the presence of 5% strength palladium-on-charcoal, ~3~

-the mixture is filtered and the fil-trate is freeze-dried.
This gives 2-acetamido-3-0-~L-l--(D-1 carbamoyl-3-carbo~Yypropyl) carbamo~l-2-hydroxye~thyl]-carbamoylmethyl}-2-deoxy-D-glucose, The s-tarting material can be prepared as follows:
3.26 g of L-serine-D-isoglutamine tert.-butyl ester-hydrochloride and 2.6 g of 2-ethoxy-N-ethoxycarbonyl-1,2 dlhydroquinoline (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 150 ml of tetrahydrofurane and the mix-ture is left to stand for 20 hours at room temperature.
After evaporating off the solvent, the residue is dissolved in 9/1 chloroform/methanol, the solution is washed with water, ice-cold 2 N hydroch]oric acid, water, a saturated solution of sodium bicarbonate and water and filtered and the filtrate is freed from the solven-t. This gives the tert.-butyl ester of benzyl-2-acetamido-3-0-~[L-l-(D-l~carbamoyl-3-carboxypropyl)-carbamoyl-2-hydroxy-ethyl~-carbamoylmethyl~-2-deoxy-a-D-glucopyranoside~
By means of mild acid hydrolysis, as described in Example 11, -the tert.-butyl ester is hydrolysed and benzyl-2-acetamido-3-0-~L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoyl-2-hydroxy-ethyl]-carbamoylmethyl3-2-deoxy-a-D-glucopyranoside is obtained.
Exam~le 37 A 5% strength solution of benzyl-2-acetamido-3-0-~L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoylbutyl]-carbamoyl-methyl3-2-deoxy-a-D-glucopyranoside in 1/1 methanol/water is c~

hydrogena-ted in the presence of 5,~ strength palladium-on-charcoal, ~the mix-ture is filtered and the filtra-te is evapor-ated. The residue is dissoLved in distilled water and freeze-dried~ This gives 2-acetamido-3-0-~[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoylbutyl]-carbamoylmethyl~-2-deoxy-D-glucose.
The s-tarting material can be prepared as follows:
3.28 g of L-norvaline-D-isoglutamine tert. bu-tyl ester-hydrochloride and 2.6 g of 2-ethoxy-N-ethoxycarbonyl-l,Z-dihydroquinoline (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 lef-t 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-butyl~-carbamoyl-methyl}-2-deoxy-a-D-glucopyranoside.
The ~tert.-butyl ester is split by mild acid hydrolysis.
Exam_le 38 A solution of 8.3 g of the benzyl ester of benzyl-3-0-~L-1-(D-l-carbamoyl-3-carboxypropyl)-carbamoylethyl]-car-bamoylmethyl3-2-deoxy-2-stearoylamido-~-D-glucopyranoside in 200 ml of glacial acetic acid is hydrogenated under normal pressure and at room temperature in the presence of 5~ streng~th palladium-on-charcoal. After working up, 3-0-~[L-1-(D-l-carbamoyl-3-carboxypropyl)-carbamoylethyl]-carbamoylmethyl3-2-deoxy-2-stearoylamido-D-glucose is ob~ained as a white powder.

- 6~+ -The starting ma-terial used can be prepared as follows:
0.75 g of pract. sodium hydride is added to a solu-tion of 6.2 g of benzyl-476-0-benzylidene-2-deoxy 2-stearoylamido-a-D-glucopyranoside in 150 ml of N3N-dimethylformamide and the mixture is stirred at 40 for 1-2 hours. The mixture is now cooled to -5, 2.2 ml of ethyl bromoacetate are added and -the mix-ture is stirred for 4 hours a-t room temperature.
~fter adding 10 ml of glacial acetic acid and 400 ml of distilled water~ the mixture is extracted with chloro~orm and the or~an_c phase is washed with water, dried and evapora-ted to dryness.
The resulting ethyl ester of benzyl-4,6-0-benzylidene-3-0-carboxymethyl-2-deoxy-2-stearoylamido~a-D glucopyranoside is crystallised from ethanol; melting point 151-154, [a]D
+94 +1 (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 of benzyl-4,6-0-benzyli-dene-3-0-carboxymethyl-2-deoxy-2-stearoylamido-a-D-gluco-pyranoside in 334 ml of methanol and 334 ml of -te-trahydrofurane 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-4J6-0-benzylidene-3-0-carboxymethyl-2-deoxy-2-stearoylamido-a-D-glucopyranoside.lH20 melts at 225-242 (with decomposition), [a]D =t45 +l (chloroform) c = 1.097).
20 mmols of L-alanine-D-isoglutamine benzyl ester-trifluoroace-tate and 4.95 g (20 mmols) of 2-etho~y-N-ethoxy-carbonyl-1,2-dihydroquinoline (EED0) are added to a solution d ~

o~ 14.4 ~ (20 mmols) of sodium benzyl-4,6-0-benzylidene-3-0-carboxymethyl 2-deoxy-2-stearoylamido-~-D glucopyranoside monohydra-te in 120 ml of tetrahydrof`urane and the mixture is left to s-tand for 20 hours at room -temperature and evaporated to dryness. The residue is dissolved in chloroform and this solution is washed with water, ice cold 2 N hydrochloric acid, water, a satura-ted solution of sodium bicarbonate and water, dried over magnesium sulphate and evapora~ted. The product, that is to say the benzyl es-ter of benzyl-4,6-0-benzylidene-3-0-~[L-l-(D-l-carbamoyl-3-carboxyprop~fl)-carbamoyl-ethyl]-carbamoylmethyl}-2~deoxy-2-stearoylamido-D-glucopyrano-side is recrystallised from ethanol; [a]D = +62 +1 (N,N-dimethylformamide, c = 1.148).
Mild acid hydrolysis of this benzylidene derivative in 65,b strength ace-tic acid leads to the benzyl ester of benzyl-3-0-~L-l-(D-l-carbamoyl~3-carboxypropyl)-carbamoylethyl]-carbamoylme~thyl~-2-deoxy-2-stearoylamido-a-D-glucopyranoside;
melting point 188-189, [~]DO =+59 +1 (N,N-dimethylformamide, c = 1.022).
Exam~le 39 A 5% s-trength aqueous solution of benzyl-2-acetamido-3-0-~[L-1-(3-1-carbamoyl-3-carboxypropyl)-carbamoyl-2-methylpropyli- carbamoylmethyl~-2-deoxy-~-D-glucopyranoside is hydrogenated in the presence of 5% strength palladium-on-charcoal and filtered and the fil-trate is lyophilised. This gives 2-acetamido-3-0-~[L-l--D-l-carb2moyl-3-carbox~ropyl)-car-bamoyl-2-me~thyl-propyl]-carbamoylmethyl3-2-deoxy-D-glucose.

The s~tarting material can be prepared as follows:
3.38 g of L-valine-D-isoglutamine tert.-bu-tyl ester-hydrochloride and 2.6 g of 2-ethoxy-N-ethox~Jcarbonyl-1,2-dihydroquinoline are added to a solution of 3.7 g of benzyl-2-acetamido-3-0-carboxymethyl-2-deoxy-~-D-glucopyranoside and 1038 ml of triethylamine in 100 ml of tetrahydrofurane and the mix-ture is left to s-tand for 24 hours a-t 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-carbamo-~1-3-carbo~propyl)-carbamoyl-2-methy].propyl ]-carbamoylmethyl¦-2-deoxy-~D-glucopyranoside, which is hydrolysed under mild acid condi-tions to give benzyl-2-acetamido-3-0-~[L-l-(D-l-carbamoyl-3-carboxypropyl)~carbamoyl-2-methylpropyl]-carbamoylmethy13-2-deoxy-~-D-glucopyranoside.
Example 40 A 5% strength solution of benzyl-2-acetamido-3-0-~[L- ~
l-(D-l-carbamoyl-3-carboxypropyl)-carbamoyl-3-me~thylbu~tyll-carb2m-oylmethyl¦-2-deoxy-~-D-glucopyranoside in 50% strength aqueous methanol is hydrogenated in the presence of 5% palladium-on-charcoal and ~the mixture is filtered and the filtrate is evap-orated. The residue is dissolved in distilled water, the solution is filtered ag2in and the filtrate is freeze-driedO
This gives 2-acetamido-3-0-~[L-l-(D-l-carbamoyl-3-carboxypropyl~-carbamoyl-3 -methylbutyl~-carbamoylme~thyl~-2-deoxy-D-glucose.
The starting material used can be prepared as follows:
3.52 g OI L-leucine-D-isoglutamine tert.-butyl ester-h~drochloride and 2.6 g of 2-ethoxy-N-e~thoxycarbonyl-1,2-dihy(lroquinoline (EEDQ) are added to a solution of 3.7 g of benzy]-2-acetamido-3-0-carboxyme~thyl-2-deoxy--a-~-glucopyranoside and 1.58 ml o~ trie-thylamine in ]00 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-ace-tamido-3-0-~L-l-(~
carbamoyl-3-carboxypropyl)-carbamoyl-3-methylbutyl]-carbamoyl-methyl¦-2-deoxy-a-D-glucopyranoside, and mild acid hydrolysis of this leads to ben~yl-2-acetamido-3-0-~[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoyl-3-me-thylbutyl]-carbamoylmethyl3-2-deoxy-a-D-glucopyranoside.
Exam~le 4l A 5% strength solution of benzyl~2-acetamido-3-0-~[L-l-(D-l-carbamoyl-3-ca-rboxypropyl)-carbamoylpropyl]-carbamoyl-methyl~-2-deoxy-~-D-glucopyranoside in 50~'0 strength aqueous methanol is hydrogenated under normal pressure and at room temperature in the presence of 5% strength palladium-on-charcoal, the catalyst is filtered off and the fil-trate is evapora-ted. The residue is dissolved in distilled water and the solution is freeze-dried. This gives 2-acetamido-3-0-~L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoylpropyl]-carbamoylmethyl3-2-deoxy-D-glucose.
The star~ting material can be prepared as follows:
3.24 g of N-a-L-aminobutyryl-D-isoglutamine tert.-butyl ester-hyd-rochloride and 2.6 g of 2-e-thoxy-N-ethoxycarbonyl-1,2-dihydroquinoline are added to a solution of 3.7 g of benzyl-2-acetamido-3-0-carboxymethyl-2-deoxy-a-D-glucopyrano-side and 1038 ml of triethylamine in 100 ml of tetrahydro-furane and after ~the mixture has s-tood for 2L~ hours at room tempera-ture i-t is worked up as described in Example 36. The resulting -ter-t.-butyl ester of benzyl-2-acetamido-3-0-l~L-]-~D-l-carbamoyl-3-carboxypropyl)-carbamoylpropyl~-carbamoyl-methyl~-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 ) ~carbamoylpropyl ~ -carbamo~
methyl~-2-deoxy-~-D-glucopyranoside.
Exam-~le l-~2 A 5~0 strength solution of benzyl-2-acetamido-3-0-~[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoylphenyl-methyl]-carbamoylmethyl~-2-deoxy-~-D-glucopyranoside in 50% strength aqueous methanol is hydrogenated under normal pressure and at room temperature in the presence of 5% strength palladium-on charcoal, the catalyst is filtered off and the filtra-te is evaporated. The residue is dissolved in distilled water and the solution is freeze-dried and the residue is 2-acetamido-3-0-~[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoylphenyl-methyl~-carbamoylmethyl~-2-deo~y-D-glucose.
The starting material can be prepared as follows:
3.72 g of l-phenylglycine-D-isoglutamine tert.-butyl ester-hydrochloride and 2.6 g of 2-ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline ~EEDQ) are added to a solution of 3.7 g of benzyl-2-acetamido-3-0-carboxymethyl-2-deoxy-~-D-glucopyrano-side and 1.38 ml of triethylamine in 100 ml of tetrahydrofurane and the mix-t-ure is left to stand at room temperature for 24 hours and worked up as described in Example 36. This glves the ter-t.-bu~tyl es-ter of benzyl-2-acetarnido-3-0-~ (D-l-carbamoyl-3~carboxypropyl)-carbamoyl-phenylmethyl]-carbamoyl-me-thyl~-2-deoxy-a-D-glucopyranoside, which is hydrolysed urlder mild acid conditions to give benzyl-2-acetamido-3-0-~ CL_1-(D~l-carbamoyl-3-carboxypropyl)-carbamoylphenylmethyl]-carbamoylmethylJ-2-deoxy-a-D-glucopyranoside.
Exam~le L~
50 ml of distilled water are added dropwise in the course of 15 minutes to a solution of 3.5 g of benzyl-2-acetamido-3-0-~L-l-(D-1,3-bis-carbamoylpropyl)-carbamoylethyl]-carbamoyl-methyl3-2-deoxy-4,6-0-isopropylidene-~-D-glucopyranoside in 50 ml of glacial acetic acid, at 50, whilst stirring, and the mix-ture is stirre~ 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-~ CL~ 1-(D-1,3-bis-carbamoyl-propyl)-carbamoylethy ]-carbamoylmethyl3-2-deoxy-~-D-gluco-pyranoside which has an Ca]D of -44 +1 (N,N-dimethylformamide7 c = 0.989).
The s-tarting 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 -D-g~ucopyranoside ln 300 ml of N7N-dimethylformamide, whils-t stirring and with external cooling and the exclusion of moistureO A solution of 20 ml of isopropenyl methyl e-ther in 60 ml of N~N-dimethyl-formamide is now added dropwise in the course of 1 hour and the mixture is stirred at room temperature for a further two hours and rendered alkaline wi-th trie-thylamine. After evapora-ting off the solvent, benzyl-2-acetamido-2-deoxy-4,6-0-isopropylidene-~-D-glucopyranoside is crystal]ised from e-thylace-tate; melting point 194, [a]D = -104 +1 (chloro-forrn, c = 0.850).
4.95 g of prac-t. sodium hydride are added to a solu-tion of 36.2 g of benzyl-2-ace~amido-2-deoxy-4,6-0-isopropylidene-~-D-glucopyranoside in 400 ml of acetonitrile and 100 ml of N,N-dime-thylformamide and the mixture is stirred for 2 hours at 40. This mixture is now cooled to -10C, 17.2 ml of e-thyl bromoacetate are added and the mixture is s-tirred for 30 min-utes at 0. 40 ml of ethanol are now added and ~the mix-ture is neutralised with glacial acetic acid and evaporated to dryness. The residue is partitioned between ether and distilled water and the ether phase is washed with water, dried over magnesium sulphate and evaporated~ The product, that is to say the ethyl ester of benzyl-2-ace-tamido-3-0-carboxymethyl-2-deoxy-4,6-0-isopropylidene-~-D-glucopyranoside is crystallised from ether; melting point 93~94~ D0 =
-49 +1 (CHC13, c _ loOOl)~
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-acetamldo-3-0-carbamoylmethyl-2-deoxy-4,6-0-isopropylidene-~-D-gluco-pyranoside in 70 ml of methanol. After the hydrolysis of -the ester has ended, 7.5 ml of 1 N hydrochloric acid are added and the mixture is evapora-ted to dryness. The resulting sodi~n 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-e-thoxy-M-ethoxycarbonyl-~L,2-dihydroquinoline (EEDQ) are added. After standing for 24 hours at room temperature, the mixture is evapora~ted to dr-yness, the residue is dissolved in water and the solu-tion 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-~ CL-l-(D-1,3-bis-carbamoylpropyl)-carbamoylethyl]-carbamoylmethy]3-2-deoxy-4,6-0-isopropylidene-~-D-glucopyranoside.
Exam~le 44 $0 ml of distilled water are added dropwise to a solu-tion of 5.1 g of the dimethyl ester of benzyl-2-acetamido-3-0-~[L-l-(D-1,3-bis-carboxypropyl)-carbamoylethyl~-carbamoyl-methyl~-2-deoxy-4,6-0-isopropylidene-~ D-glucopyranoside in 120 ml of glacial acetic acid, at 50, whilst stirring, and the mix-ture is stirred for one hour at this temperature. After evaporating off the acetic acid, the dimethyl ester of benzyl-2-ace-tamido-3-0-~[L-l-(D-1,3-bis-carboxypropyl)~carbamoylethyl]-carbamoylmethylJ-2-deoxy-~-D-glucopyranoside is obtained.
The starting material used can be prepared as follows:
6.56 g of the ethyl ester of benzyl-2-acetamido-3-0-carboxymethyl-2-deoxy-4,6-0-isopropylidene-~-D-glucopyranoside are hydrolysed as described in Example 43 with sodium hydroxide solution and the reaction product is subjected to a condensa-tion reaction with 4.24 g of L-alanyl-D-glutamic acid dimethyl ester-hydrochloride in ~the presence of 3.72 g of 2-ethoxy-N-ethox~carbonyl-1,2~dihydroquinoline. After evaporating off the solven~t, the residue is taken up in chloroform and -the solution is ~Jashed wi-th water, ice-cold 2 N hydrochloric acid9 water, a saturated solution of sodium bicarbonate and water, dried ov-er magnesium sulphate and evaporated to drynessO The residue is the dimethyl es~ter of benzyl-2-acetamido-3-0-~L-l-(D-1,3-bis-carboxypropyl)-carbamoylethyl]-carbamoylmethyl3-2-deoxy~4,6-0-isopropylidene-~-D-glucopyranoside.
Exam~le 45 A 5% streng-th solution of the benzyl ester of benzyl-2-ace-tamido-3-0-~[L-l-(D l-carbamoylmethylcarbamoyl-3-carboxy-propyl)-carbamoylethyl]-carbamoylmethyl¦-2-deoxy-a-D-gluco-pyranoside 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 ca~talyst is filtered off and ~the filtrate is evaporated. The residue is now dissolved in distilled water and further hydrogenatedA
After the theoretical amount of hydrogen has been taken up, the catalyst is filtered off and the filtrate is freeze-dried.
This gives 2-acetamido-3-0-~[L-1-(D-1-carbamoylmethylcarbamoyl-3-carboxypropyl)-carbamoylethyl]-carbamoylme~thyl3-2-deoxy-D-glucose in ~the form of a white powder.
The starting ma-terial can be prepared as follows:
4.8 g of N-tert.-butoxycarbonyl-L-alanine-D-glutamine (benzyl es~ter)-glycine-amide monohydrate are dissolved in a b ~

mix-ture of 5 ml of trifluoroacetic acid and ~ ml o~ 1,2-dichloro--ethane and -the solution is lef-t to stand for 16 hours at room tempera-ture, wi-th ~the exclusion of moisture. This solution is dilu~ted with 50 ml of tetrahydrofurane, cooled in an ice ba~th and neu-tralised with trie~thylamine. Af-ter adding a solution of 3.7 g of benzyl-2-ace-tamido-3-carboxymethyl-2-deoxy-~-D--glucopyranoside and 1.38 ml of triethylamine in ]00 ml of ~tetrahydrofurane, 2.6 g of 2-ethoxy-~-ethoxycarbonyl-1,2-di~ydroquinoline are added to the whole and -the mixture is left to stand for 24 hours at room temperature and worked up as des-cribed in Example 36. This gives the benzyl ester of benzyl-2 acetamido-3-0-~L-l-(D-1-carbamoylmethylcarbamoyl-3-carboxy-propyl)-carbamoylethyl]-carbamoylmethyl~-2-deoxy-~-D-gluco-pyranoside.
Exam~le 46 -A solution of 5.2 g of the benzyl ester of benzyl-4,6-0-benzylidene-3-0-~[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbam-oylethyl]-carbamoylmethyl3-2-deoxy-2-p-tolylsulphonylamino-~-D-glucopyranoside in 120 ml of 70,/0 streng~th aqueous acetic acid is hydrogena~ted under normal pressure and at room temperature in the presence of palladium-on-charcoal. After the theo-retical amount of hydrogen has been ta~en up, the catalyst is filtered off and the filtrate is evaporated to dryness. The residue is dissolved in 30 ml of distilled water and the solu-tion is freeze-dried. This gives 3-0-~[L-l-(D-l-carbamoyl-3-carboxypropyl)-carbamoylethyl]-carbamoylmethyl~-2-deoxy-2-p-tolylsulphon~lamino-D-glucose in the form of a white powder.

The starting material used can be prepared as follows:
A solu-tion of 3.1 g of p-~toluenesulphonyl chloride in 30 Iml of methylene chloride is added dropwise, at 5, -to a solu-tion of 6.0 g of benzyl-2-amino~4,6-0-benzylidene-3-0-carboxy-methyl-3-deoxy-a-D-glucopyranoside in 100 ml of methylene chloride and 6.3 ml of triethylamine and -the mixture is left to stand for 16 hours at room tempera-ture. This solu-tion is then washed with wa-ter, 2 N sodium hydroxide solution, water, cold 2 r~ hydrochloric acid and water, dried over mag~esium sul~
phate and evaporated to dryness. Theresidue isex-tracted severl-times with ether and dried and is benzyl-4,6-0-benzylidene-3-0-carbo~.y-methyl-2-deoxy-2-tolylsulphonylamino-a-D-glucopyranoside which has a me]ting point of 185-187C and an [a~D of -~75 +1 (chloroform, c = 0.855).

10 mmols of L-alanyl-D-isoglutamine benzyl ester-tri-fluoroacetate and 2.5 g of 2- ethoxy-N-ethoxycarbonyl-1,2-dihydroquinoline are added to a solution of 5.7 g of benzyl-4,6-0-benzylidene-3-0-carboxymethyl-2-deoxy-2-tolylsulphonyl-amino-a-D-glucopyranoside and 1.01 g of triethylamine in 80 ml of N,N-dimeth~lformamide and the mixture is left to stand for 24 hours at room temperature. It is then evaporated to dry-ness, water is added to the residue and the insoluble matter is filtered off, rinsed with water and ether and dried in vacuo.
This gives the benzyl ester of benzyl-4,6-0-benzylidene-3-0-~[L-l~(D-~L-carbamoyl-3 carboxypropyl)-carbamoylethyl]-carbamoyl-methyl}-2-deoxy-2-tolylsulphonylamino-a-D-glucopyranoside in the forrrl of a white po~lder.

- 75 ~

Example 47 2-(Acetaminomethylcarbonylamino)-2-desoxy-3,0-~[L~ D-l-carbamoyl-3-carboxy-propyl)-1.-carbamoyl-ethyll-carbamoyl-methyl~ -D-glucose is obtained analogously to Example 33.
ExamT~le 48 In a manner analogous to that described before 2-tri-methylacetamido-2-desoxy-3,0- ~L-l- (D-l-carbamoyl-3-carbo~sy-propyl)-l-carbamoyl-ethyl3-carbamoylmethyl~-,~-D-glucose is ob~ained as Lyophilisate.

- 75a -

Claims (40)

THE EMBODIMENTS OF THE INVENTION IN WHICH AN EXCLUSIVE
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:

1. A glucosamine derivative of the general formula I
(I) in which X denotes a carbonyl or sulphonyl group, R denotes lower alkyl, lower alkyl substituted by hydroxyl or carboxyl, phenyl, lower alkyl phenyl, lower alkoxy phenyl, halophenyl or trifluoro-methylphenyl, R1 denotes lower alkyl or benzyl, R2 denotes hy-drogen or lower alkyl, R4 and R6 denote hydrogen, alkyl or alka-noyl, R7 denotes hydrogen, lower alkyl, hydroxymethyl, mercapto-methyl or phenyl, R8 denotes carboxyl, lower alkyl esterified carboxyl, carbamoyl, lower alkyl carbamoyl, benzyl carbamoyl or carbamoylmethyl carbamoyl, and R9 denotes carboxyl, lower alkyl esterified carboxyl, benzyl esterified carboxyl, carbamoyl and lower alkyl carbamoyl, with the proviso that the optionally substituted alkyl radical R has more than one carbon atom if X

denotes a carbonyl group and the radical R2 denotes methyl, or, if X denotes a carbonyl group, the radical R2 represents hydrogen and R8 and R9 each represent a carboxyl group, or a salt thereof.

2. A glucosamine derivative of formula I according to claim 1, in which X denotes a carbonyl radical and R, R1, R2, R4, R6, R7, R8 and R9 have the meanings defined in claim 1, or a pharmaceutically acceptable salt thereof.

3. A compound according to claim 2, in which R2 represents hydrogen or a pharmaceutically acceptable salt thereof.

4. A compound according to claim 1, of the general formula:

(II) in which R denotes lower alkyl or phenyl, R1 denotes benzyl or lower alkyl, R2 denotes hydrogen or methyl, R7 denotes hydrogen, lower alkyl or hydroxymethyl, R8 denotes carbamoyl and R9 denotes carboxyl, with the proviso that a lower alkyl radical R contains more than 1 carbon atom if R2 denotes methyl, or a salt thereof.

5. A compound according to claim 4, in which R1 represents lower alkyl, or a pharmacelltically acceptable salt thereof.

6. A compound according to claim 4 or 5, in which R2 represents hydrogen, R denotes phenyl, R7 denotes methyl and R8 and R9 represent carboxyl, carbomethoxy, carbamoyl or N-methyl-carbamoyl.

7. A compound according to claim 4 or 5, in which R2 represents hydrogen, R denotes methy] or phenyl, R7 denotes hydroxymethyl, R8 is a carbamoyl and R9 is carboxyl.

8. A compound according to claim 4 or 5, in which R2 represents hydrogen, R denotes phenyl, R7 is methyl, R9 represents carboxyl and R8 denotes N-propylcarbamoyl or carbamoylmethyl-carbamoyl.

9. A compound according to claim 4 or 5, in which R2 represents hydrogen, R and R7 denote methyl and R8 and R9 represent carbomethoxy, carbamoyl or N methyl-carbamoyl.

10. A compound according to claim 5, in which R represents phenyl, R1 denotes ethyl, R2 represents hydrogen, R7 is methyl, R8 represents carbamoyl and R9 represents carboxyl or R8 and R9 represent carbomethoxy or N-methylcarbamoyl.

- 78a -

11. A compound according to claim 4, wherein R and R7 represent methyl, R1 is benzyl, R2 represents hydrogen, R7 is methyl and R8 and R9 represent carbomethoxy or carbamoyl.

12. A glucosamine derivative according to claim 1, of the formula in which R denotes ethyl, R1 denotes benzyl, and R2 and R7 denote methyl.

13. A glucosamine derivative according to claim 1, of the formula in which R and R7 denote methyl, R1 denotes benzyl, R2 denotes hydrogen and R6 denotes stearoyl.

14. A compound according to claim 4 or 5, in which R
denotes phenyl, R2 and R7 denote methyl, R8 denotes carbamoyl and R9 denotes carboxyl.

15. A process, according to claim 4 or 5, in which R denotes phenyl, R2 denotes hydrogen, R7 denotes methyl, R8 denotes carba-moyl and R9 denotes carboxyl.

16. Benzyl-3-0-{D-1-[L-1-(D-1-carbamoyl-3-carboxypropyl)-carbamoylethyl]-carbamoyl-ethyl}-2-deoxy-2-propionylamino-.alpha.-D-glucopyranoside.

17. Benzyl-3-0-{D-1[L-1-(D-1-carbamoyl-3-carboxypropyl)-car-bamoylethyl]-carbamoyl-ethyl}-2-deoxy-2-propionylamino-.alpha.-D-gluco-pyranoside.

18. Benzyl-2-acetamido-3-0-[L-1-(D-1-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyl-methyl-2-deoxy-6-O-stearoyl-.alpha.-D-glucopyranoside.

19. A compound according to claim 1 which is the benzyl ester of benzyl-2-acetamido-3-0-{[L-1-(D-1-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyl-methyl}-2-deoxy-.alpha.-D-glucopyra-noside.

20. Process for the manufacture of a glucosamine derivative of the general formula I
(I) in which X denotes a carbonyl or sulphonyl group, R denotes lower alkyl, lower alkyl substituted by hydroxyl or carboxyl, phenyl, lower alkyl phenyl, lower alkoxy phenyl, halophenyl or trifluoro-methylphenyl, R1 denotes hydrogen, lower alkyl or benzyl, R2 denotes hydrogen or lower alkyl, R4 and R6 denote hydrogen, alkyl or alkanoyl, R7 denotes hydrogen, lower alkyl, hydroxymethyl, mercaptomethyl or phenyl, R8 denotes carboxyl, lower alkyl esteri-fied carboxyl, carbamoyl, lower alkyl carbamoyl, benzyl carbamoyl or carbamoylmethyl carbamoyl, and R9 denotes carboxyl, lower alkyl esterified carboxyl, benzyl esterified carboxyl, carbamoyl and lower alkyl carbamoyl, with the proviso that the optionally sub-stituted alkyl radical R has more than one carbon atom if X
denotes a carbonyl group and the radical R2 denotes methyl, or, if X denotes a carbonyl group, the radical R2 represents hydrogen and R8 and R9 each represents a carboxyl group, and its salts, wherein (a) a compound of the formula (VII) in which X, R, R?, R? and R? possess the abovementioned meanings of X, R, R1, R4 and R6 provided that any hydroxyl groups which may be present therein are protected by a protective group which can be split off easily, is reacted with a compound of the formula (VIII) in which Z represents a reactive esterified hydroxyl group and R?, R? and R? have the abovementioned meanings of R7, R8 and R9 provided that any protective groups which may be present therein are split off hydrogenolytically or by acid hydrolysis; or (b) for the manufacture of a compound of the formula I wherein R4 and R6 represent hydrogen, the oxazoline and dioxolane ring in a compound of the formula (IX) in which R, R2, R?, R? and R? possess the abovementioned meaning and R5 represents an alkylidene or cycloalkylidene radical, is split open under acid conditions and any protective groups which may be present are split off hydrogenolytically or by acid hydrolysis and the X-R radical is introduced into the amino group in the 2-position of the sugar molecule if this group has been liberated and, if desired, a resulting compound is converted into its salts.

21. Process according to claim 20, wherein in process variant (b) the oxazoline and dioxolane rings are split open by means of trifluoroacetic acid in a suitable solvent.

22. A process according to claim 20, wherein in the starting materials X denotes a carbonyl radical and R, R?, R2, R?, R?, R?, R?, and R? have the meanings defined in claim 20.

23. A process according to claim 22, in which R2 represents hydrogen.

24. A process according to claim 20, in which in the starting materials, X, if present, denotes carbonyl, R denotes lower alkyl or phenyl, R?, if present, denotes a group or lower alkyl, R2 denotes hydrogen or methyl, R?, if present, denotes a hydroxy-protecting group, R? denotes hydrogen, lower alkyl or protected hydroxymethyl, R? denotes a hydroxy-protecting group, R?
denotes carbamoyl and R? denotes protected carboxyl, with the proviso that a lower alkyl radical R contains more than 1 carbon atom if R2 denotes methyl.

25. A process according to claim 24, in which R?
represents a hydroxy-protecting group.

26. A process according to claim 24, in which R?, if present, represents a hydroxy-protecting group, R2 represents hydrogen, R denotes phenyl, R7 denotes methyl. and R? and R? represent protected carboxyl, carbomethoxy, carbamoyl or N-methyl-carbamoyl.

27. A process according to claim 24, in which R?, if present, represents a hydroxy-protecting group, R2 represents hydrogen, R denotes methyl or phenyl, R? denotes protected hydroxymethyl, R? is a carbamoyl and R? is protected carboxyl.

28. A process according to claim 24, in which R?, if present, represents a hydroxy-protecting group, R2 represents hydrogen, R denotes phenyl, R? is methyl, R? represents protected carboxyl and R? denotes N-propylcarbamoyl or carbamoylmethylcarbamoyl.

29. A process according to claim 24, in which R?, if present, represents a hydroxy-protecting group, R2 represents hydrogen, R and R? denote methyl and R? and R? represent carbomethoxy, carbamoyl or N-methyl-carbamoyl.

30. A process according to claim 24, in which R represents phenyl, R? denotes ethyl, R2 represents hydrogen, R? is methyl, R? represents carbamoyl and R? represents protected carboxyl or R? and R? represent carbomethoxy or N-methylcarbamoyl.

31. A process according to claim 24, wherein R and R/
represent methyl, R? is benzyl, R2 represents hydrogen, R? is methyl and R? and R? represent carbomethoxy or carbamoyl.

32. A process according to claim 20, in which in the starting materials, X denotes carbonyl R denotes ethyl, R?
denotes benzyl, R2 and R? denote methyl, R? and R? denote hydroxy protecting groups and R? denotes carbamoyl.

33. A process according to claim 20, in which in the starting materials, X denotes carbonyl, R and R? denote methyl, R? denotes benzyl, R2 denotes hydrogen, R? denotes a hydroxy-protecting group, R? denotes stearoyl, R? denotes carbamoyl and R? denotes protected carboxyl.

34. A process according to claim 20, in which in the starting materials, X, if present, denotes carbonyl, R and R? denote methyl, R?, if present, denotes a hydroxy-protective group, R2 denotes ethyl, R? and R?, if present, denote hydroxy-protective groups, R? denotes carbamoyl and R?
denotes protected carboxyl.

35. A process according to claim 24, in which R denotes phenyl, R?, if present, denotes a hydroxy-protective group, R2 and R? denote methyl, R? denotes carbamoyl and R? denotes protected carboxyl.

36. A process according to claim 20, in which in the starting materials, X, if present, denotes carbonyl, R denotes phenyl, R? denotes a hydroxy-protective group, R2 denotes ethyl, R? and R? both denote hydroxy-protective groups, R? denotes methyl, R? denotes carbamoyl and R? denotes protected carboxyl.

37. A process according to claim 24, in which R denotes phenyl, R?, if present, denotes a hydroxy-protective group, R2 denotes hydrogen, R? denotes methyl, R? denotes carbamoyl and R? denotes carboxyl.

38. A process according to claim 20, wherein the oxazoline and dioxolane rings in a compound of the formula in which R5 denotes isopropylidene, R denotes phenyl, R2 denotes ethyl, R? denotes methyl, R? denotes carbamoyl and R?
denotes benzyloxycarbonyl, are split open under acid conditions and the benzyl ester group is split off under acid conditions or hydrogenolytically.

39. A process according to claim 20, wherein a compound of the formula in which R5 denotes isopropylidene, R denotes phenyl, R2 denotes hydrogen, R? denotes methyl, R? denotes carbamoyl and R?
denotes benzyloxycarbonyl, is split open under acid conditions and the benzyl ester group is split off under acid conditions or hydrogenolytically.

40. A pharmaceutical composition for stimulating immunity comprising an immunity stimulating amount of a glucosamine deriva-tive of the formula 1 as claimed in any one of claims 2 to 4 or a pharmaceutically acceptable salt thereof together with a pharmaceutical carrier.