NO860625L - ACYLATED HEXOS DERIVATIVES AND PROCEDURES FOR PREPARING THEREOF. - Google Patents

Description

The present invention relates to 1-O-acylated muramyl peptide and analogous D-mannose or D-galactose derivatives, methods for their production, pharmaceutical preparations containing these derivatives, and use of the preparations as medicines.

The invention relates in particular to the compounds of formula I,

in which the hexose moiety is derived from D-glucose, D-mannose or D-galactose, n stands for 0 or 1, and Ri stands for lower alkanoyl or benzoyl, R<2> stands for lower alkyl or phenyl, R<3>, R <5> and R<7> stand independently of each other for hydrogen or lower alkyl, or R^ and R^ together stand for

trimethylene and R<7> stand for hydrogen, R^ and R° independently stand for hydrogen, lower alkanoyl or benzoyl, R^ stands for hydrogen or unsubstituted or lower alkyl substituted with phenyl, hydroxy, mercapto or lower alkylthio, R<9>and R<12> independently stands for hydroxy, amino or lower alkoxy, R<10> stands for hydrogen, carboxy or lower alkoxycarbonyl and RH stands for hydrogen or unsubstituted or with amino, lower alkanoylamino, hydroxy, guanidino, lower alkanoyloxy, 2-benzyloxycarbonylaminoethylsulfinyl . and R<l>^ stands for hydrogen, R^ stands for methyl, R<9> for amino and R<l2> for hydroxy and in which the radicals Ri R<4> and R^ all have the same meaning, Ri, R^ and R° different from

acetyl and butyryl, when R<2> stands for phenyl and simultaneously R<3> for hydrogen and that R<1>, R4 and R° are different from acetyl when R<2> and R<3> both stand for methyl, and salts of such compounds with at least one salt-forming group.

The above assumption excludes N-benzoyl-1,4,6-tri-0-butyryl-desmethylmuramyl-L-alanyl-D-isoglutamine, N-benzoyl-1,4,6-tri-0-acetyl-desmethylmuramyl-L- alanyl-D-isoglutamine and N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-alanyl-D-isoglutamine from the compounds of formula I.

Preferably, the hexose moiety is derived from D-glucose.

In the case of asymmetric substitution, the configuration of the atoms C-R3 is C-R<8>resp. C-CO-R<9>(D), (L) resp. (D), as given in formula I. The configuration of C-R!! is in the case of asymmetric substitution (L) or (D), preferably (L).

Lower alkanoyl R<*>, R<4> and R° are especially C2_6_alkanoyl, e.g. n-hexa-noyl, mainly C2-5-alkanoyl and preferably C2-4-alkanoyl, e.g. propionyl, butyryl or preferably acetyl.

Lower alkyl R<2> is preferably C1_4-, especially C1_2-alkyl.

Lower alkyl R<3>, R^ and R<7> are preferably C1-3-alkyl, especially methyl.

Unsubstituted lower alkyl R<8> or R^ is preferably C1-4-alkyl, e.g. ethyl, isopropyl, 2-methylpropyl, sec-butyl or especially methyl.

With phenyl, hydroxy, mercapto or lower alkylthio, such as especially methylthio, substituted lower alkyl R<8> is preferably correspondingly substituted C1~2~ alkyl, e.g. benzyl, hydroxyethyl, 1-hydroxyethyl, mercaptomethyl or 2-methylthioethyl.

Lower oxy R<9> or R<12> is preferably C 1-4 alkoxy, e.g. methoxy, n-butyloxy or tert-butyloxy.

Lower oxycarbonyl R 1 is preferably alkoxycarbonyl with up to 5 C atoms, e.g. methoxycarbonyl, n-butyloxycarbonyl or tert-butyloxycarbonyl.

With hydroxy or lower alkanoyloxy substituted lower alkyl RH is particularly correspondingly substituted C1_2-alkyl, e.g. hydroxymethyl, 1-hydroxyethyl, lower alkanoyloxymethyl or 1-loweralkanoyloxyethyl.

With amino or lower alkanoylamino substituted lower alkyl RH is preferably 4-amino-n-butyl or 4-lower alkanoylamino-n-butyl.

With guanidino substituted lower alkyl RH is preferably 3-guanidino-n-propyl.

With 2-benzyloxycarbonylamino-ethyl-sulfinyl or -sulfonyl, or with 2-lower oxycarbonylaminoethyl-sulfinyl or -sulfonyl substituted lower alkyl RH is preferably correspondingly substituted methyl, e.g. C 5 H 5 -CH 2 -O-C(=O)-NH-CH 2 -S(=O)-CH 2 -.

The general terms used above and in the following preferably have the following meanings: Residues designated by the prefix "lower" contain up to and including 7, especially up to and including 4 carbon atoms.

Halogen is in particular chlorine or bromine, further fluorine or iodine.

Salt-forming groups in a compound of formula I are acidic groups, e.g. free carboxyl groups, or basic groups, such as especially free amino groups. Depending on the type of the salt-forming group, the compounds of formula I form metal or ammonium salts or acid addition salts. Salts of a compound of formula I are preferably pharmaceutically usable and non-toxic, e.g. alkali metal or alkaline earth metal salts, e.g. sodium, potassium, magnesium or calcium salts, or salts with ammonia or suitable organic amines, whereby primarily aliphatic, cycloaliphatic, cycloaliphatic-aliphatic or araliphatic primary, secondary or tertiary mono-, di- or polyamines, as well as heterocyclic bases are obtained in terms of salt formation, such as lower alkyl amines, e.g. triethylamine, hydroxyl lower alkylamines, e.g. 2-hydroxyethylamine, bis-(2-hydroxyethyl)amine, 2-hydroxyethyldiethylamine or tri-(2-hydroxyethyl)amine, basic aliphatic esters of carboxylic acids, e.g. 4-amino-benzoic acid-2-diethylamino-ethyl ester, lower alkylene amines, e.g. 1-ethylpiperidine, cycloalkylamine, e.g. dicyclohexylamine, or benzylamines, e.g. N,N'-dibenzylethylenediamine, further bases of the pyridine type, e.g. pyridine, collidine or quinoline. Compounds of formula I with at least one basic group can form acid addition salts, e.g. with inorganic acids, such as hydrochloric acid, sulfuric acid or phosphoric acid, or with suitable carboxylic or sulphonic acids, e.g. trifluoroacetic acid, as well as with amino acids, such as arginine and lysine. In the presence of several acidic or basic groups, mono- or poly-salts can be formed. Compounds of formula I with an acid, e.g. free carboxyl group, and a free basic, e.g. an amino group can also exist in the form of internal salts, i.e. in doubly ionic form, or part of the molecule can exist as an internal salt and another part as a normal salt.

Pharmaceutically unsuitable salts can also be used for isolation and purification. Nevertheless, only pharmaceutically usable, non-toxic salts, which are therefore preferred, are used therapeutically.

From the French Patent Application No. 74 22 909 with the publication number

2,292,486, the muramyl peptide of the type for the compound N-acetyl-muramyl-L-alanyl-D-isoglutamine ("MDP") is known. These compounds are described as immunological adjuvants, i.e. they can be used in admixture with vaccine substances to improve the vaccine result. Own investigations, e.g. with mice infected with influenza viruses, have shown that N-acetyl-muramyl-L-alanyl-D-isoglutamine per se, i.e. without the addition of vaccine substances, is ineffective in the prophylaxis and therapy of viral infections. Nor has the applicability of structurally simpler muramyl peptides and their analogues against viral infections been described so far.

The present invention is based on the task of providing structurally relatively simple and consequently relatively easy-to-produce muramyl peptide derivatives, which, when administered to warm-blooded animals, are very effective when it comes to the prophylaxis and therapy of viral infections.

According to the invention, it has surprisingly been found that the above-mentioned compounds of formula I and their pharmaceutically usable salts are excellently suitable both for prophylaxis and also for therapy of viral diseases, this is shown e.g. in the case of animal experiments, as exemplified in the example section. In these animal experiments, animals, such as mice or guinea pigs, are infected with a wide variety of virus species in a dose that is fatal for all or the great majority of untreated (control) animals, e.g. LDgO-90»°§The course of infection in the untreated control animals is observed for comparison with the animals that have been treated with one of the above-mentioned compounds or a salt thereof before, simultaneously with or after the infection.

Hereby it turns out that a prophylactic effect upon administration of the compounds of formula I occurs already several days until some, e.g. four weeks before the infection, and a therapeutic effect still occurs when administered several days, e.g. one week, after the infection.

In the above-mentioned experiment on mice, the compounds of formula I are effective already in the dose range between 0.0001 mg/kg and 0.1 mg/kg.

Also noteworthy is the broad viral spectrum against which the above-mentioned compounds are effective.

The compounds of formula I can in particular be used for prophylaxis and therapy in diseases caused by the viruses discussed in more detail below [for nomenclature see J.L. Melnick, Prog. with. Virol, 26^214-232

(1980) and 28, 208-221 (1982)]: DNA viruses with cubic symmetry and bare nucleocapsid, DNA viruses with an enveloped virion as well as RNA viruses with cubic and those with helical symmetry for the capsid.

Preferably, the compounds of formula I are used in the case of DNA viruses with an enveloped virion and cubic symmetry for the capsid, in the case of RNA viruses with cubic symmetry for the capsid and bare virion and in the case of RNA viruses with helical symmetry for the capsid, in which the nucleocapsidome envelope ling is located at the surface membrane, but also in the case of adenoviruses, poxviruses and coronaviruses, such as human coronaviruses in particular. First of all, the compounds of formula I are used in the case of herpesviruses, picornaviruses and myxoviruses, but also in the case of mastadenoviruses, such as in particular human adenoviruses, in the case of chordopoxviruses, as mainly orthopoxviruses, as in particular e.g. vacciniavirus, in the case of reoviruses, primarily (especially human) rotaviruses, as well as in the case of caliciviruses and rhabdoviruses, primarily vesiculoviruses in humans as well as horses, bulls and pigs.

The compounds of formula I are mainly used in the case of oc-herpesviruses, such as varicellavirus, e.g. human varicella-zoster viruses, rhinoviruses, cardioviruses and orthomyxoviruses, but also in the case of p-herpesviruses, such as especially human cytomegaloviruses, in the case of aphthous viruses, primarily aphthoviruses from ungulates, mainly from bulls, as well as in the case of paramyxoviruses, primarily pneumoviruses, e.g. respiratory syncytial viruses in humans, and further morbilliviruses or paramyxoviruses, such as parainfluenza viruses, e.g. human parainfluenza viruses, including sendaiviruses as well as in cases of arboviruses or vesiculoviruses, e.g. vesicular stomatitis viruses.

First of all, the compounds of formula I are used in cases of simplex viruses, e.g. human herpes simplex viruses of type 1 and 2, in the case of human encephalomyocarditis viruses, in the case of influenza viruses such as mainly influenza A and influenza B viruses, in the case of vaccinia and parainfluenza viruses and especially in the case of the viruses mentioned in the examples.

The compounds of formula I can be used for prophylaxis and therapy in viral infections, especially for warm-blooded animals including humans, by using them orally or parenterally, primarily together with suitable auxiliary or carrier substances. Preferably they are applied to the mucosa, e.g. intranasally, rectally, vaginally or conjunctivally, or orally. However, the antiviral effect also occurs by other delivery routes, e.g. subcutaneously, intravenously, intramuscularly or by application to the normal skin.

The dosage of the active substance depends, among other things, on the type of warm-blooded animal, the organism's state of defence, the method of administration and the type of virus. The dose-effect relationship is relatively weakly pronounced.

For prevention, a one-time dose of va. 0.01 mg to approx. 10 mg, preferably 0.05 to 1 mg, e.g. 0.2 mg of active substance for a warm-blooded animal with a body weight of approx. 70 kg, e.g. a human being. The prophylactic effect of this dose lasts for several weeks. If necessary, e.g. in times of increased risk of infection, the supply of this dose can<g>be increased.

The therapeutic dose for warm-blooded animals with a body weight of approx. 70 kg is between 0.1 mg and 25 mg, preferably between 01 and 1 mg, e.g. at 0.5 mg, especially by oral administration. The dosage for topical, especially intranasal administration is a factor of 10 lower. If necessary, the administration of the compound of formula I can be repeated until an improvement in the disease state occurs. Often, however, a single supply is sufficient.

The compounds of formula I also have antitumor properties. This depends on the ability of the connections, e.g. incorporated in multilamellar liposomes or in phosphate-buffered physiological saline solution (PBS), to activate macrophages in such a way that these body-specific defense cells are able to kill tumor cells (cytotoxicity) or prevent their growth (cytostasis). The induction of tumoricids and tumoristatic alveolar macrophages in rats in vitro and in situ allows e.g. show in the following experiment: Alveolar macrophages are obtained by washing the lungs with culture medium. These macrophages are activated either by injection of the test substance into the rat (intravenous or intranasal in situ activation) or by a 24-hour pre-incubation with a compound of the formula I in a CO 2 -incubator (in vitro activation). The macrophages activated in this way are then incubated with tumor cells for a further 72 hours.

To measure the tumoricidal activity of the macrophages, the tumor cells are labeled before the 72 hour incubation with<125>i_j0doc|eoxyuridine. After washing away the radioactivity released by the lysed tumor cells, the non-killed tumor cells can be measured on the basis of the remaining radioactivity.

To determine the tumoristatic activity of macrophages, <3>H-thymidine is added to the culture 8 hours before the end of the 72 hour incubation and then -thymidine incorporation into the tumor cells is measured. In vitro, the substances both dissolved in PBS and also incorporated in liposomes already in doses of 20 nanograms/0.2 ml culture can induce tumoricidal rat alveolar macrophages. In rats, a single intravenous administration of the compounds incorporated in liposomes at a dose of 160 µg/animal causes an induction of tumoricide and tumoristatic alveolar macrophages. Furthermore, a single intranasal administration of the substances in PBS in a dose of 25 µg/rat induces tumoricidal alveolar macrophages.

The compounds of formula I can therefore also be used on warm-blooded animals including humans for therapy in tumor diseases, especially e.g. to avoid the formation of metastases, e.g. by surgical removal of the primary tumor.

Preferred are compounds of formula I, in which the hexose part is derived from D2-glucose or D-mannose, n stands for 0, R<*>, R<4> and R^ stand independently of each other for C2-5-alkanoyl or -benzoyl, R<2> stands for (4.4-alkyl or phenyl, R<3>, R<5> and R<7> stand independently for hydrogen or methyl, or R^ and R<8> together stand for trimethylene and R<7>represents hydrogen, R<8>represents hydrogen, Cj_4-alkyl or Cj_2~alkyl substituted with phenyl, hydroxy, mercapto or methylthio, R<9>and R<12>represent independently of each other hydroxy, amino or lower alkoxy, and R<lO> stands for hydrogen, carboxy or lower alkoxycarbonyl, and salts of such compounds with at least one salt-forming group with the exception of N-benzoyl-1,4,6-tri-0-butyryl-desmethylmuramyl-L- alanyl-D-isoglutamine, N-benzoyl-1,4,6-tri-0-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine and N-acetyl-1,4,6-tri-0-acetyl-muramyl- L-alanyl-D-isoglutamine and salts thereof.

Particularly preferred are compounds of formula I, in which the hexose part is derived from D-glucose, n stands for 1, R^, R<4> and R^ stand independently of each other for <C>2-5-alkanoyl or benzoyl, R<2> stands for <C>i_4~alkyl or phenyl, R<3>, R<5>and R<7> independently stand for hydrogen or methyl, or R<5> and R<8> together stand for trimethylene and R <7> for hydrogen, R<8> stands for hydrogen, Cj_4-alkyl or Cj_2-alkyl substituted with phenyl, hydroxy, mercapto or methylthio, R<9> and R^<2> independently of each other stand for hydroxy, amino or lower alkoxy, R<lO> stands for hydrogen, carboxy or lower alkoxycarbonyl and RH stands for unsubstituted or with amino, guanidino, lower alkanoyloxy, 2-benzyloxycarbonylaminoethyl-sulfinyl, 2-benzyloxycarbonylaminoethyl-sulfonyl, 2-lower oxycarbonylaminoethyl-sulfinyl or 2-lower oxycarbonylaminoethyl-sulfonyl substituted lower alkyl, and salts of such compounds with at least one salt-forming group.

Very preferred are compounds of formula I in which the hexose part is derived from D-glucose, n stands for 0 or 1, and R^ stands for C2_5-alkanoyl or benzoyl, R<2> stands for Cj_4-alkyl or phenyl, R<3> stands for hydrogen or<C>j_4-alkyl, R4 stands for hydrogen,<C>2_5~alkanoyl or benzoyl, R^ stands for hydrogen, methyl or together with Rg for trimethylene, R<6>stands for C2_5~alkanoyl or benzoyl , R<7>represents hydrogen or methyl, R<8>represents Ci_4-alkyl or together with R<5>trimethylene, R<9>represents hydroxy, lower alkoxy or amino, R<lO>represents hydrogen or carboxy, RH stands for unsubstituted or with amino, guanidino or. 2-benzyloxycarbonylaminoethylsulfinyl substituted C1_4-alkyl and R<i2>represents hydroxy, lower alkoxy or amino, and salts of such compounds with at least one salt-forming group with the exception of N-benzoyl-1,4,6-tri-0-butyryl -desmethylmuramyl-L-alanyl-D-isoglutamine, N-benzoyl-1,4,6-tri-0-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine and N-acetyl-1,4,6-tri-0 -acetyl - muramyl-L-alanyl-D-isoglutamine and salts thereof.

Particularly preferred are compounds of formula I in which the hexose part is derived from D-glucose, n stands for 0 or 1, Ri stands for C2-5-alkanoyl or benzoyl, R<2> stands for Cj-2-alkyl or phenyl, R<3> stands for hydrogen or methyl, R<4> stands for hydrogen, C2_5-alkanoyl or benzoyl, R^ stands for hydrogen, R° stands for C2_5~alkanoyl or benzoyl, R<7> stands for

hydrogen, R<8>represents methyl, ethyl or 2-propyl, R<9>represents hydroxy, amino or<C>i_4~ alkoxy, Rl<O>represents hydrogen, RH represents methyl or 4-amino- n-butyl and R<i2>represents hydroxy, amino or C1_4~alkoxy, and salts of such compounds with at least one salt-forming group with

except for N-benzoyl-1,4,6-tri-O-butyryl-desmethylmuramyl-L-alanyl-D-isoglutamine, N-benzoyl-1,4,6-tri-O-acetyl-desmethylmuramyl-L-alanyl -D-isoglutamine and N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-alanyl-D-isoglutamine and salts thereof.

Particularly preferred are the compounds of formula mentioned in the examples

IN.

Particularly preferred are also the above-mentioned compounds of formula I in which Ri is different from acetyl and butyryl and/or R<2> is different from methyl and phenyl and/or R<4> is different from acetyl and butyryl and/or R< 5>is different from hydrogen and/or R° is different from acetyl and butyryl and/or R<7>is different from hydrogen and/or R<8>is different from methyl and/or R<9>is different from amino and /or R<10>is different from hydrogen and/or n is different from 0 and/or R<12>is different from hydroxy, and salts of such compounds with at least one salt-forming group.

Examples of such compounds can be mentioned the compounds in which R<8> stands for C2_4~alkyl or, together with R<5>, for trimethylene.

Most preferred are, due to their outstanding pharmacological action, the above-mentioned compounds of formula I in which at least one of the residues R<9> and R<i2> stands for lower alkoxy and the other of the residues R<9> and R<l2 >stands for hydroxy, amino or lower alkoxy, and/or in which n stands for 1 and RH is different from hydrogen, and the other substituents have the meanings mentioned above, and salts of such compounds with at least one salt-forming group.

The compounds of formula I and salts of such compounds with at least one salt-forming group are prepared in a manner known per se:

They are produced e.g. in that one

a) reacts a compound of formula II

in which at least one of the radicals R<1a>, R<2a> and, for the preparation of compounds of formula I, in which R<4> and R^ have the meanings given above with the exception of hydrogen, R<4a> and R<6a >stands for hydrogen, and the rest of these residues have the meaning of R<1>, the group R<2->C(=0)-, R<4> respectively. R<6>, and the rest of the substituents have the meanings indicated above, whereby a compound of formula II present free functional groups, with the exception of the groups which will participate in the reaction, can be protected if necessary with easily cleavable protective groups, with an acylating agent which transfers the acyl residue Ri, r<2->C(=0)-, R<4>or R° to be introduced and, if necessary, cleaves off protecting groups present, or

b) reacts with a compound of formula III

wherein the substituents have the meanings indicated above, whereby free

OH groups OR<4>and OR<0> are protected by easily cleavable protecting groups, or a reactive derivative thereof with a compound of formula IV

in which X stands for a reactive esterified hydroxy group, and the other substituents have the meanings mentioned above, whereby in a compound of formula IV present free functional groups, with the exception of X, are protected if necessary by means of easily cleavable protecting groups, and if necessary cleaves the protecting group present, or,

c) reacts a compound of formula V

in which q, r, s and t independently stand for 0 or 1 and in which

the substituents have the meanings mentioned above, whereby in a compound of formula V present free functional groups, with the exception of the groups which will participate in the reaction, are, if necessary, protected with

easily cleavable protecting groups, or a reactive carboxylic acid derivative thereof, with a compound of formula VI

in which u, v and x independently of each other stand for 0 or 1 and the other symbols and substituents have the meanings mentioned above, whereby in a compound of formula VI present free functional groups, with the exception of the groups which will participate in the reaction, if necessary is protected by easily removable protecting groups, whereby u, v and x stand for 1, when q and t stand for 0 in the reaction partner of formula V, or u stands for 0 and v and x stand for 1 when q stands for 1 and t stands for 0, or u and v stand for 0 and x stands for 1 when q, r and t stand for 1 and s stands for 0, or (for the preparation of compounds of formula I in which n stands for 1) u and x stands for 0, when q, r, s and t stand for 1, or a reactive derivative thereof, and if necessary cleaves off the protective groups present, or d) for the preparation of a compound of formula I, in which R<9>has one of the meanings given above except for hydroxy, and the other substituents have the meanings given above, ami derates or esterifies a compound of formula VII

in which R<1:*> stands for carboxy, and in which the other substituents have the meanings indicated above, whereby in a compound of formula VII present free functional groups with the exception of R<13> can, if necessary, be protected by means of easily removable protective groups, or a reactive carboxylic acid derivative thereof, and if necessary removes any protective groups present, or

e) in a compound of formula I, in which at least one of the residues OR<4>, OR^, R<8>, C(=0)-R9, R10, R<11> and C(=0)-R< 12>available in protected form, split off

those protecting groups which do not correspond to the definition of the desired end product,

and, if desired, after carrying out one of the methods a-e) transfers an obtained compound of formula I with at least one salt-forming group to its salt or converts an obtained salt of a compound of formula I to the free compound, and/or separates an obtained isomer mixture.

Methods a, c, d and e are preferred.

The implementation of the above-mentioned method variants shall be described in more detail below:

Procedure a:

Method a) is preferably used to introduce an acyl residue Rj, R4 and/or R5. This preferably starts from compounds of formula II, in which the residue R<2a> stands for the group R<2->C(=0).

For the preparation of compounds of formula I, where R<4> and/or R° stands for hydrogen, one preferably starts from compounds of formula II in which R<4a> and/or R<6a> stands for an easily or regioselectively cleavable hydroxy protecting group, whereby R<4a> and R^<a> can optionally also together stand for a bivalent hydroxy protecting group.

In cases with acylating agents that transfer large, bulky acyl residues, e.g. benzoyl, the selective acylation of hydroxyl groups in the 11- and 6-positions on the sugar part also succeeds with suitable reaction management, without a protection of the 4-OH group being imperatively required.

In a compound of formula II any free functional groups present, which are preferably protected with easily removable protective groups, are furthermore especially free hydroxy or mercapto in the residue R<8>, free carboxy R<9->C(=0)-, R< 10> or R<12->C(=0)- as well as free amino, hydroxy or guanidino in the residue R^.

Protection groups, as well as their introduction and separation are e.g. described in "Protective Groups in Organic Chemistry", Plenum Press, London, New York 1973, and in "Methoden der organischen Chemie", Houben-Weyl, 4th edition, volume 15/1, Georg-Thieme-Verlag, Stuttgart 1974 as well as in Theodora W. Greene, "Protective Groups in Organic Synthesis", John Wiley & Sons, New York 1981. Characteristic of protective groups is that they are easily cleavable, i.e. that the cleavage takes place without unwanted side reactions, e.g. solvolytic, reductive, photolytic or also under physiological conditions.

Hydroxy protecting groups are e.g. acyl residues, which optionally, such as e.g. with halogen, substituted lower alkanoyl, such as 2,2-dichloroacetyl, or acyl residues of carboxylic acid half-esters, especially tert-butyloxycarbonyl, optionally substituted benzyloxycarbonyl, e.g. 4-nitro-benzyloxycarbonyl, or diphenylmethoxycarbonyl, or 2-halo-lower oxycarbonyl, such as 2,2,2-trichloroethoxycarbonyl, further trityl or formyl, or organic silyl or stannyl residues, further easily cleavable etheric groups, such as tert.-lower alkyl, e.g. tert-butyl, 2-oxa- or 2-thia-aliphatic or -cycloaliphatic hydrocarbon residues, primarily

1-lower oxyalkyl or 1-lower alkylthio-lower alkyl, e.g. methoxymethyl, 1-methoxyethyl, 1-ethoxyethyl, methylthiomethyl, 1-methylthioethyl or 1-ethylthioethyl, or 2-oxa- or 2-thiacycloalkyl with 5-6 ring atoms, e.g. tetrahydrofuryl or 2-tetrahydropyranyl or corresponding thia analogs, as well as optionally substituted 1-phenyl lower alkyl, such as optionally substituted benzyl or diphenylmethyl, whereby as substituents on the phenyl residues e.g. halogen, such as chlorine, lower alkoxy, such as methoxy and/or nitro are mentioned. Two neighboring hydroxy groups, e.g. both hydroxy groups in the 4- and 6-position on the sugar part can also be protected with an optional, e.g. with a phenyl radical, substituted methylene radical or a cycloalkylidene, e.g. with lower alkylidene, such as especially isopropylidene or with benzylidene.

Carboxyl groups are usually protected in esterified form, whereby such ester groupings are easily cleavable under mild conditions. Carboxyl groups protected in this way contain, as esterifying groups, primarily branched in the 1-position or suitably substituted lower alkyl groups in the 1- or 2-position. Preferred carboxyl groups present in esterified form are among others tert.-lower oxycarbonyl, e.g. tert-butyloxycarbonyl, arylmethoxycarbonyl with one or two aryl residues, whereby these are made up of possibly e.g. with lower alkyl, such as tert.-lower alkyl, e.g. tert-butyl, lower alkoxy, such as methoxy, hydroxy, halogen, e.g. chlorine and/or nitro, mono- or polysubstituted phenyl residues, which optionally, e.g. as mentioned above, substituted benzyloxycarbonyl, e.g. 4-methoxybenzyloxycarbonyl or 4-nitrobenzyloxycarbonyl, or optionally, e.g. as mentioned above, substituted diphenylmethoxycarbonyl, e.g. diphenylmethoxycarbonyl or di-(4-methoxyphenyl)methoxycarbonyl, 1-lower oxycarbonyl, such as methoxy-methoxycarbonyl, 1-methoxyethoxycarbonyl or 1-ethoxymethoxycarbonyl,

1-lower alkylthiolower oxycarbonyl, such as 1-methylthiomethoxycarbonyl or 1-ethylthioethoxycarbonyl, aroylmethoxycarbonyl, in which the aroyl group optionally constitutes e.g. with halogen, such as bromine, substituted benzoyl, e.g. phenacyloxycarbonyl, 2-halolower oxycarbonyl, e.g. 2,2,2-trichloroethoxycarbonyl, 2-bromomethoxycarbonyl or 2-iodoethoxycarbonyl, or 2-(trisubstituted silyl)ethoxycarbonyl, in which the substituents independently of each other stand for optionally substituted, e.g. with lower alkyl, lower alkoxy, aryl, halogen and/or nitro substituted, aliphatic, araliphatic, cycloaliphatic or aromatic hydrocarbon residue, as corresponding, optionally substituted lower alkyl, phenyl lower alkyl, cycloalkyl or phenyl, e.g. 2-trilower alkylsilylethoxycarbonyl, 2-trimethylsilylethoxycarbonyl or 2-(di-n-butyl-methyl-silyl)-ethoxycarbonyl, or 2-triarylsilylethoxycarbonyl, such as 2-triphenylsilylethoxycarbonyl.

The organic silyl or stannyl radicals mentioned above and below preferably contain lower alkyl, especially methyl, as substituents on the silicon or tin atoms. Relevant silyl or stannyl groups are primarily trilower alkylsilyl, especially trimethylsilyl, further dimethyl-tert-butylsilyl, or similarly substituted stannyl, e.g. tri-n-butylstannyl.

Preferred protected carboxyl groups are tert.-lower oxycarbonyl, such as tert.-butoxycarbonyl, and above all possibly e.g. as mentioned above, substituted benzyloxycarbonyl, such as 4-nitrobenzyloxycarbonyl, or diphenylmethoxycarbonyl, especially 2-(trimethylsilyl)ethoxycarbonyl.

A protected amino group can e.g. present in the form of an easily cleavable acylamino, arylmethylamino, etherified mercaptoamino, 2-acyl-lower alk-1-en-yl-amino, silyl or stannylamino group or as an azido group.

In a relevant acylamino group, acyl is, for example, the acyl residue from an organic carboxylic acid with e.g. up to 18 carbon atoms, especially one optionally, e.g. with halogen or aryl, substituted alk ank yearbook syl sy re or optionally, e.g. with halogen, lower alkoxy or nitro substituted benzoic acid, or a carboxylic acid half ester. Such acyl groups are, for example, lower alkanoyl, such as formyl, acetyl or propionyl, halolower alkanoyl, such as 2-haloacetyl, especially 2-chloro-, 2-bromo-, 2-iodo-, 2,2,2-trifluoro- or 2,2,2 -trichloroacetyl, optionally, e.g. with halogen, lower alkoxy or nitro substituted benzoyl, e.g. benzoyl, 4-chlorobenzoyl, 4-methoxybenzoyl or 4-nitrobenzoyl, or in the 1-position on the lower alkyl radical branched or in the 1- or 2-position suitably substituted lower alkoxycarbonyl, especially tert.-lower oxycarbonyl, e.g. tert-butyloxycarbonyl, arylmethoxycarbonyl with one or two aryl residues which preferably optionally e.g. with lower alkyl, especially tert.-lower alkyl, such as tert.-butyl, lower alkoxy, such as methoxy, hydroxy, halogen, e.g. chlorine and/or nitro, mono- or polysubstituted phenyl, such as optionally substituted benzyloxycarbonyl, e.g. 4-nitro-benzyloxycarbonyl, or substituted diphenylmethoxycarbonyl, e.g. benzhydryloxycarbonyl or di-(4-methoxyphenyl)-methoxycarbonyl, aroylmethoxycarbonyl in which the aroyl group preferably constitutes, if necessary, e.g. with halogen, such as bromine, substituted benzoyl, e.g. phenylacyloxycarbonyl, 2-halolower oxycarbonyl, e.g. 2,2,2-trichloroethoxycarbonyl, 2-bromomethoxycarbonyl or 2-iodoethoxycarbonyl, or 2-(trisubstituted silyl)ethoxycarbonyl, in which the substituents independently of each other stand for optionally substituted, e.g. with lower alkyl, lower alkoxy, aryl, halogen or nitro substituted, aliphatic, araliphatic, cycloaliphatic or aromatic hydrocarbon residues with up to 15 C atoms, as optionally substituted lower alkyl, phenyl lower alkyl, cycloalkyl or phenyl, e.g. 2-trilower alkylsilylethoxycarbonyl, such as 2-trimethylsilylethoxycarbonyl or 2-(di-n-butylmethylsilyl)-ethoxycarbonyl or 2-triarylsilylethoxycarbonyl, such as 2-triphenylsilylethoxycarbonyl.

Other silyl residues that come into question as amino protecting groups are also corresponding residues of organic phosphoric, phosphonic or phosphinic acids, such as dilaverealkylphosphoryl, e.g. dimethylphosphoryl, diethylphosphoryl, di-n-propylphosphoryl or diisopropylphosphoryl, dicycloalkylphosphoryl, e.g. dicyclohexylphosphoryl, optionally substituted diphenylphosphoryl, e.g. diphenylphosphoryl, possibly e.g. with nitro substituted di-(phenyl lower alkyl) phosphoryl, e.g. dibenzylphosphoryl or di-(4-nitrobenzyl)-phosphoryl, optionally substituted phenyloxyphenyl-phosphonyl, e.g. phenyloxyphenyl-phosphonyl, dilaverealkylphosphinyl, e.g. diethylphosphinyl, or optionally substituted diphenylphosphinyl, e.g. diphenylphosphinyl.

In an arylmethylamino group, which constitutes a mono-, di- or especially triarylmethylamino group, the acyl residues are especially optionally substituted phenyl residues. Such groups are, for example, benzyl, diphenylmethyl and especially tritylamino.

An etherified mercapto group in an amino group protected by such a residue

is primarily arylthio or arylloweralkylthio, in which aryl is particularly optional, e.g. with lower alkyl, such as methyl or tert-butyl, lower alkoxy, such as methoxy, halogen such as chlorine, and/or nitro substituted phenyl. A relevant amino protecting group is e.g. 4-nitrophenylthio.

In a 2-acyl-lower alk-1-en-yl residue that can be used as an amino protecting group, acyl is e.g. the corresponding residue of a lower alkanecarboxylic acid, an optionally, e.g. with lower alkyl such as methyl or tert.-butyl, lower alkoxy such as methoxy, halogen such as chlorine, and/or nitro substituted benzoic acid, or in particular a carboxylic acid moiety, such as a carboxylic acid lower alkyl moiety. Relevant protecting groups are primarily 1-lower alkanoyl-prop-1-en-2-yl, e.g. 1-acetyl-prop-1-en-2-yl, or 1-lower oxycarbonyl-prop-1-en-2-yl, e.g. 1-ethoxycarbonyl-prop-1-en-2-yl.

An amino group can also be protected in protonated form; corresponding anions are primarily anions from strong inorganic acids, such as from halogen hydrogen acids, e.g. the chlorine or bromine anion, or from organic sulfonic acids, such as p-toluenesulfonic acid.

Preferred amino protecting groups are acyl residues from carboxylic acid half-esters, especially tert-butyloxycarbonyl, possibly e.g. as indicated, substituted benzyloxycarbonyl, e.g. 4-nitro-benzyloxycarbonyl, or diphenylmethoxycarbonyl, or 2-halo-lower oxycarbonyl, such as 2,2,2-trichloroethoxycarbonyl, further trityl or formyl.

A mercapto group such as e.g. in cysteine, can be particularly protected by S-alkylation with optionally substituted alkyl residues, thioacetal formation, S-acylation or by providing asymmetric disulfide groupings. Preferred mercapto-protecting groups are e.g. optionally in the phenyl residue, e.g. with methoxy or nitro, substituted benzyl, such as 4-methoxybenzyl, optionally in the phenyl part, e.g. with mercapto, substituted diphenylmethyl, such as 4,4'-dimethoxydiphenylmethyl, triphenylmethyl, trimethylsilyl, benzylthiomethyl, tetrahydropyranyl, acylaminomethyl, benzoyl, benzyloxycarbonyl or aminocarbonyl, such as ethylaminocarbonyl.

An acylating agent that transfers the residue R<1>, r<2->C(=0)-, R<4>or R<6> is in particular the corresponding carboxylic acid, i.e. Ri-OH, R<2->COOH, R<4->OH or R6 -OH, or preferably a reactive acid derivative thereof, whereby the activation of the carboxylic acid used as acylating agent can also take place in situ in the presence of the compound of formula II.

Activated carboxylic acid derivatives that can be used as acylating agents are primarily reactive activated esters or reactive anhydrides, further reactive cyclic amides.

Activated esters of acids are especially at the point of attachment of the esterifying residue unsaturated esters, e.g. of the vinyl ester type, such as actual vinyl ester (which can, for example, be obtained by transesterification of a corresponding ester with vinyl acetate; the process with activated vinyl esters), carbamoyl vinyl ester (which can, for example, be obtained by treating the corresponding acid with an isoxazolium reagent; the 1,2-oxazolium or Woodward method), or 1-lower oxy vinyl ester (which can be obtained, for example, by treating the corresponding acid with a lower alkoxyacetylene; the ethoxyacetylene method), or esters of the amidino type, as N,N'-disubstituted amidinoesters (which can, for example, be obtained by treating the corresponding acid with a suitable N,N'-disubstituted carbodiimide, e.g. N,N'-dicyclohexylcarbodiimide; the carbodiimide method), or N,N-disubstituted amidino ester (which can for example be obtained by treating the corresponding acid with an N,N-disubstituted cyanamide; the cyanamide method), suitable aryl ester, especially with electron-withdrawing substituents suitable substituted phenyl ester (s if one e.g. can be obtained by treating the corresponding acid with a suitable substituted phenol, e.g. 4-nitrophenol, 4-methylsulfonylphenol, 2,4,5-trichlorophenol, 2,3,4,5,6-pentachlorophenol or 4-phenyldiazophenol, in the presence of a condensing agent, such as N,N'-dicyclohexylcarbodiimide; the activated aryl ester method), cyanomethyl ester (which can be obtained, for example, by treating the corresponding acid with chloroacetonitrile in the presence of a base; the cyanomethyl ester method), thioester, especially optionally, e.g. with nitro, substituted phenyl thioester (which can, for example, be obtained by treating the corresponding acid with optionally, e.g. with nitro, substituted thiophenols, among other things by means of the anhydride or carbodiimide method; the method with activated thiol ester), amino or amido ester (which can, for example, be obtained by treating the corresponding acid with an N-hydroxyamino or N-hydroxyamido compound, "e.g. N-hydroxysuccinimide, N-hydroxypiperidine, N -hydroxyphthalimide or 1-hydroxybenztriazole, e.g. by the anhydride or carbodiimide method; the method with activated N-hydroxy ester) or silyl ester (which can be obtained, for example, by treating the corresponding acid with a silylating agent, f .eg hexamethyldisilizane and which reacts easily with hydroxy, with not with amino groups).

Anhydrides of acids can be symmetrical or preferably mixed anhydrides of these acids, e.g. anhydrides with inorganic acids, such as acid halides, especially acid chlorides (which can be obtained, for example, by treating the corresponding acids with thionyl chloride, phosphorus pentachloride or oxalyl chloride; the acid chloride method), azides (which can be obtained, for example, from a corresponding acid ester over the corresponding hydrazide and treatment of this with nitric acid; the azide method), anhydrides with carboxylic acid half-derivatives, such as corresponding esters, e.g. carboxylic acid lower alkyl half-esters which can be obtained by treating the corresponding acids with halogen, such as chloroformic acid lower alkyl esters or with a 1-lower oxycarbonyl-2-lower oxy-1,2-dihydro-quinoline, e.g. 1-lower oxycarbonyl-2-ethoxy-1,2-dihydroquinoline; the method with mixed O-alkylcarboxylic acid anhydrides), or anhydrides with dihalogenated, especially dichlorinated phosphoric acids (which can be obtained, for example, by treating the corresponding acids with phosphorus oxychloride; the phosphorus oxychloride method), or anhydrides with organic acids, such as mixed anhydrides with organic carboxylic acids (which can, for example, be obtained by treating the corresponding acids with an optionally substituted lower alkane or phenylalkane carboxylic acid halide, e.g. phenylacetic acid, pivalic acid or trifluoroacetic acid chloride; the method with mixed carboxylic acid anhydrides) or with organic sulphonic acids (which can, for example, be obtained by treating a salt, such as an alkali metal salt, of the corresponding acid, with a suitable organic sulphonic acid halide, such as lower alkane or aryl, e.g. methane or p-toluenesulphonic acid chloride; the process with mixed sulphonic acid anhydrides), as well as symmetrical anhydrides (which can, for example, be obtained by condensation of the corresponding render the acid in the presence of a carbodiimide or of 1-diethylaminopropyne; the method with symmetrical anhydrides).

Suitable cyclic amides are especially amides with a 5-membered diazacyclic aromatic character, such as amides with imidazoles, e.g. imidazole (which can be obtained by treating the corresponding acid with N,N'-carbonyldiimidazole; the imidazolide method), or pyrazoles, e.g. 3,5-dimethyl-pyrazole (which can be obtained via the acid hydrazide for treatment with acylacetone; pyrazolid - the first method).

As mentioned, derivatives of acids, which are used as acylating agents, can also be formed in situ. Consequently, one can e.g. form N,N'-disubstituted amidino ester in situ by bringing the mixture of starting materials of formula II and the acid used as acylating agent in the presence of a suitable N,N-disubstituted carbodiimide, e.g. N,N'-dicyclohexylcarbodiimide to reaction. Furthermore, one can form the amino or amido ester of the acid used as acylating agent in the presence of the starting material of formula II to be acylated, by reacting the mixture of corresponding acid and amino starting materials in the presence of an N,N'-disubstituted carbodiimide, e.g. N,N'-dicyclohexylcarbodiimide, and an N-hydroxyamine or N-hydroxyamide, e.g. N-hydroxysuccinimide, optionally in the presence of a suitable base, e.g. 4-Dimethylaminopyridine.

The reaction can be carried out in a manner known per se, whereby the reaction conditions primarily depend on whether and how the carboxyl group in the acylating agent is activated, usually in the presence of suitable solvents or diluents or a mixture thereof, and, if necessary, in the presence of a condensation agent, which, for example, when the carboxyl group participating in the reaction is present as an anhydride, can also be an acid binding agent, during cooling or heating, e.g.

in the temperature range from approx. -30 °C to approx. +150 "C, in particular from about 0°C to +100 "C, preferably from room temperature (about +20 *C) to +70 °C, in a closed reaction vessel and/or in an atmosphere of inert gas, f .ex. nitrogen. Common condensation agents are e.g. carbodiimides, for example N,N'-diethyl-, N,N'-dipropyl-, N,N'-dicyclohexyl- or N-ethyl-N'-(3-dimethyl-aminopropyl)-carbodiimide, suitable carbonyl compounds, for example carbonyldiimidazole, or 1,2-oxazolium compounds, e.g. 2-ethyl-5-phenyl-1,2-oxazolium-3'-sulfonate and 2-tert-butyl-5-methyl-isoxazolium perchlorate, or a suitable acylamino compound, e.g. 2-ethoxy-1-ethoxycarbonyl-1,2-

dihydroquinoline. Common acid-binding condensation agents are e.g. alkali metal carbonates or hydrogen carbonates, e.g. sodium or potassium carbonate or bicarbonate (usually together with a sulphate), or organic bases, usually sterically hindered tri-lower alkylamines, e.g. N,N-diisopropyl-N-ethylamine.

The cleavage of the protecting groups, e.g. The carboxyl, amino, hydroxy or mercapto protecting groups, which do not form a component of the desired end product of formula I, take place in a manner known per se, e.g. by means of solvolysis, especially hydroxy, alcoholysis or azidolysis, or by means of reduction, especially hydrogenolysis or chemical reduction, optionally stepwise or simultaneously, whereby enzymatic methods can also be used.

Consequently, one can transfer tert.-lower oxycarbonyl or in the 2-position with an organic silyl group, or in the 1-position with lower alkoxy or lower alkylthio substituted lower alkoxycarbonyl or optionally substituted diphenylmethoxycarbonyl, e.g. by treatment with a suitable acid, such as formic acid or trifluoroacetic acid, optionally with the addition of a nucleophilic compound, such as phenol or anisole, to free carboxyl. Optionally substituted benzyloxycarbonyl can e.g. is set free by means of hydrogenolysis, i.e. by treatment with hydrogen in the presence of a metallic hydrogenation catalyst, such as a palladium catalyst. Furthermore, one can transfer suitable substituted benzyloxycarbonyl, such as 4-nitrobenzyloxycarbonyl, also by means of chemical reduction, e.g. by treatment with an alkali metal, e.g. sodium dithionite, or with a reducing number, e.g. zinc, or metal salt, such as a chromium II salt, e.g. chromium II chloride, usually in the presence of a hydrogen-releasing agent, which together with the metal can develop masking hydrogen, such as an acid, primarily a suitable carboxylic acid, which optionally, e.g. with hydroxy, substituted lower alkanecarboxylic acid, e.g. acetic acid, formic acid, glycolic acid, diphenyl-glycolic acid, lactic acid, mandelic acid, 4-chloro-mandelic acid or tartaric acid, or an alcohol or thiol, whereby water is preferably added to the carboxyl. By treatment with a reducing metal or metal salt, as described above, one can also convert 2-halo-lower oxycarbonyl (possibly after conversion of a 2-bromo-lower oxycarbonyl group to a corresponding 2-iodo-lower oxycarbonyl group) or aroylmethoxycarbonyl to free carboxyl, whereby aroylmethoxycarbonyl can optionally be cleaved by treatment with a nucleophilic, preferably salt-forming reagent, such as sodium thiophenolate or sodium iodide. Substituted 2-silyloxycarbonyl can also be transferred by treatment with a salt of hydrofluoric acid which releases fluoride anion, such as an alkali metal fluoride, e.g. sodium or potassium fluoride, in the presence of a macrocyclic polyether ("crown ether"), or with a fluoride of an organic quaternary base, such as tetra-lower alkyl ammonium fluoride or tri-lower alkyl aryl ammonium fluoride, e.g. tetraethylammonium fluoride or tetrabutylammonium fluoride, in the presence of an aprotic polar solvent, such as dimethylsulfoxide or N,N-dimethylacetamide, to free carboxyl.

Esterified carboxyl can also be split enzymatically, e.g. esterified lysine, e.g. lysine methyl ester, using trypsin.

A protected amino group is set free in a manner known per se and depending on the type of protecting group in a different manner, preferably by means of solvolysis or reduction, 2-halogen-lower oxycarbonylamino (possibly after conversion of a 2-bromo-lower oxycarbonylamino group into a 2-iodo -lower oxycarbonylamino group), aroylmethoxycarbonylamino or 4-nitrobenzyloxycarbonylamino can e.g. is decomposed by treatment with a suitable chemical reducing agent, such as zinc in the presence of a suitable carboxylic acid, such as aqueous acetic acid. However, aroylmethoxycarbonyl-amino can also be cleaved by treatment with a nucleophilic, preferably salt-forming reagent, such as sodium thiophenolate, and 4-nitrobenzyloxycarbonylamino also by treatment with an alkali metal, e.g. sodium dithionite. Optionally substituted diphenylmethoxycarbonylamino, tert.-lower oxycarbonylamino or 2-trisubstituted silylethoxycarbonylamino can be cleaved by treatment with a suitable acid, e.g. formic acid or trifluoroacetic acid, optionally substituted benzyloxycarbonylamino, e.g. by means of hydrogenolysis, i.e. by treatment with hydrogen in the presence of a suitable hydrogenation catalyst, such as a palladium catalyst, optionally substituted triarylmethylamino or formylamino, e.g. when treated with

an acid, such as mineral acid, e.g. hydrochloric acid, or an organic acid,

in e.g. formic acid, acetic acid or trifluoroacetic acid, possibly in the presence of water, and an amino group protected with an organic silyl group can e.g. is set free by means of hydrolysis or alcoholysis. One with 2-

haloacetyl, e.g. The 2-chloroacetyl protected amino group can be set free by treatment with thiourea in the presence of a base, or with a thiolate salt, such as an alkali metal thiolate, of the thiourea and subsequent solvolysis, such as alcoholysis or hydrolysis of the resulting condensation product. A 2-substituted silylethoxycarbonyl-protected amino group can also be transferred to the free amino group by treatment with a salt of hydrofluoric acid which gives off a fluoride anion, as indicated above in connection with the release of a corresponding protected carboxyl group.

In the form of an azido group, protected amino is transferred to free amino, e.g. by reduction, for example by catalytic hydrogenation with hydrogen in the presence of a hydrogenation catalyst, such as platinum oxide, palladium or Raney nickel, or also by treatment with zinc in the presence of an acid, such as acetic acid. The catalytic hydrogenation is preferably carried out in an inert solvent, such as a halogenated hydrocarbon, e.g. methylene chloride, or also in water or a mixture of water and an organic solvent, such as an alcohol or dioxane, at approx. 20 °C to 25 °C, or also during cooling or heating.

A hydroxy or mercapto group protected with a suitable acyl group, an organic silyl group or optionally substituted 1-phenyl lower alkyl is set free analogously to a corresponding protected amino group. With optionally substituted 1-phenyl lower alkyl, e.g. benzyl, protected hydroxy is preferably split off by catalytic hydrogenation, e.g. in the presence of a palladium-carbon catalyst. A with 2,2-dichloroacetyl protected hydroxy- or mercapto group is set free, e.g. by basic hydrolysis, one with tert.-lower alkyl or with a 2-oxa- or 2-thia-aliphatic or -cycloaliphatic hydrocarbon residue etherified hydroxy- or mercapto group by acidolysis,

e.g. by treatment with a mineral acid or a strong carboxylic acid, e.g. trifluoroacetic acid. Two hydroxy groups, which are protected together by means of a preferably substituted methylene group, as in the lower alkylidene, e.g. isopropylidene, cycloalkylidene, e.g. cyclohexylidene, or benzylidene, can be set free by acid solvolysis, especially in the presence of a

mineral acid or a strong organic acid. With an organic silyl residue, e.g. in trimethylsilyl, etherified hydroxy can also be set free with a salt of hydrofluoric acid which gives off a fluoride anion, e.g. tetrabutylammonium fluoride.

Preferably, the protecting groups are selected in the presence of several protected functional groups in such a way that more than one such group can be cleaved at the same time, e.g. acidolytically, as by treatment with trifluoroacetic acid or formic acid, or reductively, as by treatment with zinc and acetic acid, or with hydrogen and a hydrogenation catalyst, such as a palladium-carbon catalyst. When the desired end compounds contain protecting groups, e.g. when R 1 means with 2-benzyloxycarbonylaminoethylsulfinyl isosubstituted lower alkyl, the protective groups which are to be removed after completion of the reaction are chosen so that they can be removed regioselectively, e.g. with an organic silyl residue etherified hydroxy in the residue R<4>, R° or R<8> can be set free with fluoride, whereby a benzyloxycarbonyl protecting group elsewhere in the molecule remains intact.

The starting material of formula II, in which R<2a> stands for hydrogen, is obtained, for example for the corresponding compound in which R<2a> stands for an easily cleavable amino protecting group, e.g. benzyloxycarbonyl, by splitting off this amino protecting group, whereby e.g. benzyloxycarbonyl can be cleaved by hydrogenation in the presence of a catalyst consisting of 10% palladium on carbon.

Procedure b

A reactive derivative of a compound of formula III is e.g. a metal-oxy compound, such as can be obtained by reacting a compound of formula III with a suitable base, such as an alkali metal hydride or amide.

Reactive esterified hydroxy X is e.g. with a strong inorganic or

organic acid esterified hydroxy, as with a mineral acid, e.g. hydrohalic acid, such as hydrochloric, hydrobromic or hydroiodic acid, further sulfuric acid, or halosulphuric acid, e.g. fluorosulphuric acid, or with a strong organic sulphonic acid, as a possibly, e.g. with halogen, such as fluorine, substituted lower alkanesulfonic acid or an aromatic

sulphonic acid, e.g. an optionally lower alkyl, such as methyl, halogen, such as bromine, and/or nitro substituted benzenesulfonic acid, e.g. a methanesulfone-,

trifluoromethanesulfonic or p-toluenesulfonic acid, esterified hydroxy, preferably a chloride, bromide or iodide.

Free functional groups in a compound of formula IV, which are preferably protected by means of easily cleavable protecting groups, are in particular hydroxy, mercapto or carboxy groups.

The reaction is preferably carried out in an inert organic solution at a temperature between -30°C and +150°C, especially 0°C and +100°C, e.g. +20"C and +70°C, if necessary in the presence of an acid-binding agent.

The removal of protective groups, which are not a component of the desired end product of formula I, takes place as described in method a).

Procedure c:

A reactive carboxylic acid derivative of a compound of formula V is primarily a reactive ether, a reactive anhydride or a reactive cyclic amide, in which the carboxyl group is activated in a similar way to the reactive acylating agents mentioned in method a), and whereby the activation can also take place in situ.

Optionally present functional groups in the compounds of formulas V or VI, which are preferably protected by means of easily removable protecting groups, are in particular amino or guanidino in residue R<11>, carboxy as residue R^ and hydroxy as residue R<9> or R12 moreover also hydroxy or mercapto in residue R<8> or hydroxy in residue R^.

In a reactive derivative of a compound of formula VI, the amino group is activated, e.g. by reaction with a phosphite, e.g. diethyl chlorophosphite, 1,1-phenylene chlorophosphite, ethyl dichlorophosphite, ethylene chlorophosphite or tetraethyl pyrophosphite or by bonding to halogenocarbonyl, e.g. chlorocarbonyl, or in the form of an isocyanate group. Preferably, the reaction is carried out in such a way that a reactive carboxylic acid derivative of a compound of formula V is reacted with a compound of formula VI, in which the amino or hydroxy groups participating in the reaction are present in free form.

The reaction and the subsequent removal of the protective groups, which do not form part of the desired end product, are carried out as described in method a).

The esterification of a compound of formula V, in which q, r and t stand for 1, can also be carried out using an esterification agent mentioned under method b).

Procedure d:

Free functional groups present in a compound of formula VII, which are preferably protected by means of easily cleavable protecting groups, are in particular free carboxy R^Og -C(=O)-R^<2>. When it is appropriate, free hydroxy OR<4> and OR<6>, free hydroxy or mercapto in the residue R<8> or free amino, hydroxy or guanidino in the residue R^ are also protected. The protecting groups and their removal are described in method a).

A reactive carboxylic acid derivative of a compound of formula VII is preferably a reactive ester, a reactive anhydride or a reactive cyclic amide, in which the carboxyl group is activated in a similar way to the reactive acylating agents mentioned in method a), and whereby the activation can also take place in situ . Preferably, the reaction is carried out in such a way that a reactive carboxylic acid derivative is reacted

of a compound of formula VII with a compound H-R<9a>, in which R<9a> has the above-mentioned meaning for R<9> with the exception of hydroxy.

Alternatively, a compound of formula VII with a free carboxyl group can be esterified by reaction with a reactive derivative of the alcohol desired as esterification component.

in

Suitable agents for esterification are e.g. corresponding diazo compounds, such as optionally substituted diazolaveralkanes, e.g. diazomethane, diazoethane or diazo-n-butane. These reagents are used in the presence of

a suitable inert solvent, such as an aliphatic, cycloaliphatic or aromatic hydrocarbon, such as hexane, cyclohexane, benzene or toluene, a halogenated aliphatic hydrocarbon, e.g. methylene chloride or an ether, such as a dilower alkyl ether, e.g. diethyl ether, or a cyclic ether, e.g.

tetrahydrofuran or dioxane, or a solvent mixture and, depending on the diazo reagent, under cooling, at room temperature or under slight heating, further, if necessary, in a closed container and/or under an atmosphere of inert gas, e.g. nitrogen.

Further suitable agents for esterification are esters of corresponding alcohols, primarily those with strong inorganic or organic acids, such as mineral acids, e.g. hydrohalic acids, such as hydrochloric, hydrobromic or hydroiodic acid, further sulfuric acid, or halosulphuric acid, e.g. fluorosulphuric acid, or strong organic sulphonic acids which, if necessary, e.g. with halogen, such as fluorine, substituted lower alkanesulphonic acids, or aromatic sulphonic acids, such as e.g. optionally with lower alkyl, such as methyl, halogen, such as bromine, and/or nitro substituted benzenesulfonic acids, e.g. methanesulfonic, trifluoromethanesulfonic or p-toluenesulfonic acid. Such esters are, among other things, lower alkyl halides, dilower alkyl sulfates, such as dimethyl sulfate, further fluorosulfonic acid esters, such as lower alkyl esters, e.g. fluorosulfonic acid methyl ester, or optionally halogen-substituted methanesulfonic acid lower alkyl esters, e.g. trifluoromethanesulfonic acid methyl ester. They are usually used in the presence of an inert solvent, such as an optionally halogenated, such as chlorinated, aliphatic, cycloaliphatic or aromatic hydrocarbon, e.g. methylene chloride, an ester, such as dioxane or tetrahydrofuran, or a mixture thereof. Hereby, suitable condensation agents are preferably used, such as alkali metal carbonates or hydrogen carbonates, e.g. sodium or potassium carbonate or hydrogen carbonate (usually together with a sulfate), or organic bases, which are usually sterically hindered trilower alkylamines, e.g. N,N-diisopropyl-N-ethylamine (preferably together with halosulfonic acid lower alkyl esters or optionally halogen-substituted methanesulfonic acid lower alkyl esters), whereby

work is carried out during cooling, at room temperature or during heating, e.g. at temperatures from approx. -20 °C to approx. 50 °C, and, if necessary, in a closed container and/or in an atmosphere of inert gas, e.g. nitrogen.

Further agents for esterification are corresponding trisubstituted oxonium salts

(so-called "Meerwein" salts), or disubstituted carbenium or halonium salts, in which the substituents are the etheric residues, for example trilavereal alkyloxonium salts, as well as dilavereal oxycarbenium or dilaverealalkylhalonium salts, especially the corresponding salts with com-

plex, fluorine-containing acids, such as the corresponding tetrafluoroborates, hexafluorophosphates, hexafluoroantimonates, or hexachloroantimonates. Such reagents are e.g. trimethyloxonium or triethyloxonium hexafluoroantimonate, -hexachloroantimonate, -hexafluorophosphate or -tetrafluoroborate, dimethoxycarbenium hexafluorophosphate or dimethylbromonium hexafluoroantimonate. These agents are preferably used in an inert solvent, such as an ether or a halogenated hydrocarbon, e.g. diethyl ether, tetrahydrofuran or methylene chloride, or a mixture thereof, if necessary, in the presence of a base such as an organic base, e.g. a, preferably sterically hindered, tri-lower alkylamine, e.g. N,N-diisopropyl-N-ethylamine, and under cooling, at room temperature or under slight heating, e.g. at approx. -20°C to approx. 50"C, if necessary, in a closed container and/or in an atmosphere of inert gas, eg nitrogen.

A preferred embodiment of method d) is the reaction of a cesium salt of a compound of formula VII with the alcohol desired as esterification component, in which the hydroxyl group is present in reactive esterified form.

Procedure e:

A compound of formula I, in which at least one of the residues R<4>, R<6>, R<8>, R<9>(RiO, R11 and R<1>2 is present in a protected form, which does not correspond to the definition of the desired end product, is in particular a compound of formula I in which a hydroxy group OR<4>or OR<6>, a hydroxy or mercapto group in residue R<8>, a carboxyl group R<10>, an amino or hydroxy group in residue RH and/or free hydroxy R<9>or R<1>2 is protected with an easily cleavable protecting group, which is not present in the desired end product.

Easily cleavable protective groups are especially the groups mentioned in method a). The removal of the protecting group is carried out as described in procedure a).

Additional operations:

Salts of compounds of formula I with salt-forming groups can be prepared in a manner known per se. Consequently, salts of compounds of formula I with acidic groups can be prepared by reaction with a suitable base, e.g. by treatment with suitable metal compounds, such as alkali metal salts of suitable organic carboxylic acids, e.g. the sodium salt of c<-ethylcaproic acid, or with suitable inorganic alkali or alkaline earth metal salts, especially salts derived from weak and preferably volatile acids, e.g. sodium hydrogen carbonate, or with ammonia or a suitable organic amine, whereby stoichiometric amounts or only a small excess of the salt-forming agent are preferably used. Acid addition salts of compounds of formula I are obtained in the usual way, e.g. by treatment with an acid or a suitable anion exchange reagent. Internal salts of compounds of formula I, such as e.g. contains a free carboxyl group or a free amino group, can e.g. is formed by neutralizing salts, such as acid addition salts, to the isoelectric point, e.g. with weak bases, or when treated with liquid ion exchangers.

Salts can normally be transferred to the free compounds, metal and ammonium salts, e.g. by treatment with suitable acids, and acid addition salts, e.g. by treatment with a suitable basic agent.

Mixtures of isomers can be divided into the individual isomers in a manner known per se, e.g. by fractional crystallization, chromatography, etc.

The methods mentioned above included the methods for removing protective groups and the other method features

is carried out, when not stated otherwise, in a manner known per se, e.g. in the presence or absence of preferably inert solvents and diluents, if necessary, in the presence of condensing agents or catalysts, at reduced or elevated temperature, e.g. in a temperature range from approx. -20"C to approx. +150 °C, especially from approx. 0°C to approx. +70°C,

preferably from approx. +10 °C to approx. +40 °C, mainly at room temperature j in a closed container and/or in an atmosphere of inert gas, e.g.

nitrogen.

In doing so, one must take into account all the substituents that are present

itself in the molecule, if necessary, e.g. in the presence of easily hydrolyzable j residues, use particularly mild reaction conditions, such as short reaction times, use of mild acids or basic agents in low

concentrations, stoichiometric quantity ratios, choice of suitable catalysts, solvents, temperature and/or pressure conditions.

The invention also relates to those embodiments of the method in which one starts from an intermediate product obtained at any step of the method and carries out the other method steps, or interrupts the method at any step, or forms a starting material under the reaction conditions, or uses it in the form of a reactive derivative or salt. This is preferably based on starting materials which, in the method according to the invention, lead to the compounds mentioned above as particularly valuable.

New starting materials and/or intermediate products as well as methods for their production are also subject to the present invention. Preferably, one starting material is used and the reaction conditions are chosen in such a way that compounds are arrived at which in the present description are designated as particularly valuable.

The invention also relates to pharmaceutical preparations containing an effective amount of the active substance together with pharmaceutically usable carriers, which are suitable for topical use, e.g. for prophylaxis and therapy in the case of viral infections. intranasal, enteral, e.g. oral or rectal, or parenteral administration, and which may be inorganic or organic, solid or liquid. Consequently, tablets or gelatin capsules are used which contain the active substance together with diluents, e.g. lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycerin, and/or lubricants, e.g. silica, talc, stearic acid or salts thereof, such as magnesium or calcium stearate, and/or polyethylene glycol. Tablets may also contain binders, e.g. magnesium aluminum silicate, starches, such as corn, wheat or rice starch, gelatin, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone, and, if desired, explosives, e.g. starch, agar, alginic acid or a salt thereof, such as sodium alginate, and/or fizzy mixes, or adsorbents, colourants, flavorings and sweeteners. Furthermore, the pharmacologically active compounds according to the present invention can be used in the form of parenterally administrable preparations or in the form of infusion solutions. Such solutions are preferably isotonic aqueous solutions or suspensions, whereby these e.g. in the case of lyophilized preparations, which contain the active substance alone or together with a carrier material, e.g. mannitol, can be prepared before use. The pharmaceutical preparations may be sterilized and/or contain preservatives, stabilizers, wetting and/or emulsifying agents, solubility mediators, salts for regulating the osmotic pressure and/or buffers. The present pharmaceutical preparations, which if desired may contain further pharmacologically active substances, such as antibiotics, are prepared in a manner known per se, e.g. by means of conventional mixing, granulating, coating, dissolving or lyophilization methods, and contains from approx. 0.001% to 20%, especially from approx. 0.01% to approx. 10%, primarily between 0.1% and 5% of that, respectively. the active substances, whereby a concentration of active substance of less than 1% is particularly suitable for preparations to be applied topically.

As administration forms for topical application, the following are preferred: creams, ointments or pastes with an active substance content of from 0.001% to 1%, especially from 0.01% to 0.1%, e.g. 0.05%, e.g. ointments for intranasal administration or lipsticks, or aqueous solutions with an active substance content of 0.001% to 1%, especially 0.05% to 0.5%, e.g. 0.1%, preferably isotonic, sterile and physiologically tolerable solutions, e.g. eye drops, preferably in microcontainers for single use, or sprays for use in the mouth and throat.

The pharmaceutical preparations mentioned in the examples are particularly suitable.

Creams are oil-in-water emulsions that contain more than 50% water. Fatty alcohols are primarily used as oily bases, e.g. lauryl, cetyl or stearyl alcohols, fatty acids, e.g. palmitic or stearic acid, liquid to solid waxes, e.g. isopropyl myristate, wool wax or beeswax, and/or hydrocarbons, e.g. petroleum jelly (petrolatum) or paraffin oil. Emulsifiers include surfactants with predominantly hydrophilic properties, such as corresponding non-ionic emulsifiers, e.g. fatty acid esters of polyalcohols or ethylene oxide addition products thereof, such as polyglycerin fatty acid esters or polyoxyethylene sorbitan fatty acid esters ("Tweens"), further polyoxyethylene fatty alcohol ethers or fatty acid esters, or corresponding ionic emulsifiers, such as alkali metal salts of fatty alcohol sulfates, e.g. sodium lauryl sulfate, sodium cetyl sulfate or sodium stearyl sulfate, which are usually used in the presence of fatty alcohols, e.g. cetyl alcohol or stearyl alcohol. Additives to the water phase include agents that reduce the drying out of the cream, e.g. polyalcohol, such as glycerin, sorbitol, propylene glycol and/or polyethylene glycols, further preservatives, fragrances, etc.

Ointments are water-in-oil emulsions that contain up to 70%, preferably from approx. 20% to approx. 50% water or aqueous phase. The fat phase is primarily hydrocarbons, e.g. petroleum jelly, paraffin oil and/or hard paraffins in speech, which preferably contain suitable hydroxy compounds, such as fatty alcohols or esters thereof, for example, cetyl alcohol or wool wax alcohols, to improve the water-binding capacity, or wool wax. Emulsifiers are similar lipophilic substances, such as sorbitan fatty acid esters ("Spans"), e.g. sorbitan oleate and/or sorbitan isostearate. Additives to the water phase are e.g. agents that retain moisture, such as polyalcohols, e.g. glycerol, propylene glycol, sorbitol and/or polyethylene glycol, as preservatives, fragrances, etc.

Grease ointments are anhydrous and contain as a basis in particular hydrocarbons, especially paraffin, vaseline and/or liquid paraffins, further natural or partially synthetic fatty substances, e.g. coconut fatty acid triglyceride, or preferably hardened oil, e.g. hydrogenated peanut or castor oil, further fatty acid partial esters of glycerol, e.g. glycerol mono- and distearate, as well as e.g. the fatty alcohols mentioned in the context of the ointments which are capable of improving hydrogen absorption, emulsifiers and/or additives.

Pastes are creams and ointments with secretion-absorbing powder ingredients, such as metal oxides, e.g. titanium oxide or zinc oxide, further talc and/or aluminum silicates, which have the task of binding the moisture or secretions present.

Foam is administered from a pressurized container and is in aerosol form available liquid oil-in-water emulsions, whereby halogenated hydrocarbons, such as chlorofluorolower alkanes, e.g. dichlorodifluoromethane and dichlorotetrafluoroethane are used as propellants. Hydrocarbons are used as the oil phase, e.g. paraffin oil, fatty alcohols, e.g. cetyl alcohol, fatty acid ester, e.g. isopropyl myristate and/or other waxes. Emulsifiers used include mixtures of those with predominantly hydrophilic properties, such as polyoxyethylene sorbitan fatty acid esters ("Tweens") and compounds with predominantly lipophilic properties, such as sorbitan fatty acid esters ("Spans"). In addition, there are the usual additives such as preservatives etc.

Tinctures and solutions mostly have an aqueous-ethanolic base, to which, among other things, polyalcohols have been added, e.g. glycerol, glycols, and/or polyethylene glycol, as agents that retain moisture to reduce evaporation, and fattening substances, such as fatty acid esters with lower polyethylene glycols, i.e. soluble in aqueous mixtures, lipophilic substance as a replacement for the fatty substances that are extracted from the skin with the ethanol , and, if necessary, other aids and additives.

The production of the topically usable pharmaceutical preparations takes place in a manner known per se, e.g. by dissolving or suspending the active substance in the base, or, if necessary, in a part thereof. When processing the active substance as a solution, this is usually dissolved before emulsification in one of the two phases; when processed as a suspension, it is mixed after emulsification with part of the base and then with the rest of the composition.

The following examples illustrate the invention. The Rf values are measured on silica gel thin-layer plates (Company Merck, Darmstadt). The ratio between the eluents in the emulsifier mixtures used is stated in volume fractions (V/V). The concentration, c, of the substance in the solvent (solvent mixture) is, in the case of optical polarization, expressed as a percentage (weight/volume).

Abbreviations:

abs. = absolutely

Boe = tert-butyloc sy carbonyl

HV = high vacuum

i.vak. = in vacuum

Me = methyl

MeOH = methanol

PTFE = polytetrafluoroethylene, "Teflon"

RT = room temperature

Temp. = melting point

Dec. = decomposition

EXAMPLE 1

A solution of 5.77 g (7.3 mmol) 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester (a,p mixture) in 80 ml methanol-tetrahydrofuran (1:1) is hydrated for one hour at room temperature and normal pressure with 311 mg of 10% palladium carbon as catalyst. The catalyst is then filtered off and the filtrate is evaporated to dryness in a vacuum at 30 °C. The yellow residue obtained in this way is distributed three times between water-saturated n-butanol and water. The butanol phases are collected and evaporated in vacuum at 40 °C. The obtained almost colorless residue is dissolved in 100 ml of double-distilled water-tert.-butanol (1:1) and the obtained solution is filtered through a millipore filter ("Fluorpore", PTFE, 0.2 pm). The clear, colorless filtrate is then lyophilized in high vacuum.

Pure 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine (cx,<p->mixture) is obtained as a colorless powder, which contains 1.16 mol of water and 0.437 moles of tert.-butanol.

C25H38N4°14'1.16H2° '°'437(CH3)3C-°H (671.87)

Calculated: C 47.82 H 6.70 N 8.34 O 37.14 H20 3.27 (CH3)3C-OH4.82 Found: C 47.70 H 6.61 N 8.51 O 36.99 H20 3 .27 (CH3)3C-OH4.82 [a]J)° = +39.0 ± 0.1° (c = 0.682; methanol),

Rf = 0.51 (methylene chloride: methanol: water = 70:30:5).

The starting material is produced in the following way:

Step 1. 1

16.0 g (31.63 mmol) of N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine containing 0.74 mol of water is dissolved in 300 ml of methanol-dimethoxyethane (1:1) and then mixed with 9.2 g (47.4 mmol) of diphenyldiazomethane. The whole is stirred for 20 hours at room temperature. After 20, 44 and 68 hours

a further 4.6 g (23.7 mmol) of diphenyldiazomethane is added in each case. After completion of the reaction (90 hours), the red solution is evaporated in vacuo at 40°C to dryness, the red resin obtained is mixed with 300 ml of absolute diethyl ether and stirred for 1/2 hour at room temperature.

Colorless crystals are obtained which are filtered off and washed with diethyl ether.

After recrystallization from 400 ml of acetone, N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester (cx, p-mixture) is obtained in the form of colorless crystals with m.p. 188-190°C) (decomposition), which contains I, 17 moles of water.

C32H42N40irl,17H20 (679.78)

Calculated: C 56.55 H 6.60 N 8.24 O 28.63 H20 3.09

Found: C 56.67 H 6.67 N 8.43 O 28.65 H2O 3.09

[a]D° = + 7.5 ± 0.1° (c = 0.898; methanol),

Rf = 0.62 (chloroform:methanol:water = 70:30:5),

Rf = 0.62 (chloroform:methanol = 7:3).

Step 1. 2

II, 4 g (16.77 mmol) of N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester (cx, p-mixture) containing 1.17 mol of water are dissolved in 120 ml of absolute pyridine. The solution obtained is mixed with 7.6 ml (80.3 mmol) of acetic anhydride and stirred for 92 hours at room temperature. The yellowish solution is then evaporated in high vacuum at 40°C. The crude product obtained is triturated with 100 ml of diethyl ether and the resulting suspension is stirred for 2 hours at room temperature. The formed crystals are then suctioned off and washed with diethyl ether.

After recrystallization twice from acetic acid ethyl ester-isopropanol-diethyl ether (100:5:40) a first fraction of chromatographically pure 1,4,6-tri-O-acetyl-N-propionyl-desmethyl muramyl-L-alanyl-D- isoglutamine-benzhydryl ester (cx,p mixture) in the form of colorless crystals with m.p. 165-166°C.

The mother liquor from the recrystallization is collected and evaporated in vacuo. The obtained residue is purified by column chromatography on 600 g of silica gel ("type 60", pure, Merck; 0.063-0.2 mm) in the system chloroform-ethanol (9:1) (10 ml fractions).

Fractions 217-310 are collected and evaporated in vacuo. A second fraction of chromatographically pure 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester (cx, p-mixture) is obtained, which together with the first fraction is dissolved in 200 ml absolute methanol. The slightly cloudy solution obtained is then filtered through a millipore filter ("Fluoropore", PTFE, 0.2 pm) and the clear filtrate is evaporated in vacuo at 40 "C. The residue is then dissolved in 50 ml of absolute ethyl acetate which has previously been filtered through a millipore filter ("Fluoropore", PTFE, 0.2 pm) and crystallized by adding 20 ml of absolute diethyl ether which has also been filtered through a millipore filter, and washed with filtered, absolute diethyl ether. You get 1,4,5-tri-O- acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester (cx, p-mixture) in the form of colorless crystals of mp 165-166° C., which still contain 0.27 mol of water.

C38H48N4O14-0.27H2O (789.68)

Calculated: C 57.80 H 6.22 N 7.10 O 28.91 H20 0.62

Found: C 57.91 H 6.20 N 7.11 O 28.77 H20 0.62

[cx]j)° = + 32.7 ± 2.1° (c = 0.486, methanol),

Rf = 0.28 (chloroform:methanol = 9:1)

Rf = 0.70 (chloroform:methanol = 4:1)

A further product is obtained from fractions 105-140 by the column chromatography described above.

The yellow residue (0.55 g, foam) obtained after evaporation in vacuo is dissolved while hot in cyclohexane-diethyl ether-methylene chloride (10:30:5) and the solution obtained is cooled to room temperature. This separates a yellow oil which, after decanting off the solvent, is stirred for 1/2 hour with 50 ml of absolute diethyl ether. The crystals obtained are filtered off and washed with absolute diethyl ether.

The crystals are then dissolved in 100 ml tert.-butanol-water (1:1). The solution obtained is filtered through a millipore filter ("Fluoropore", PTFE, 0.2 µm) and lyophilized.

La,4,6-tri-0-acetyl-2-deoxy-2-propionylamino-D-mannos-3-0-yl-acetyl-L-alanyl-D-isoglutamine-benzhydryl ester is obtained as a colorless powder containing 1, 11 moles of water.

C38H48N40l4'UlH20 (804.81)

Calculated: C 56.71 H 6.31 N 6.96 O 30.03 H20 2.48

Found: C 56.94 H 6.36 N 7.22 O 30.15 H20 2.48

[cx]D° = + 5.5 ± 2.7° (c = 0.365; methanol)

Rf = 0.49 (chloroform:methanol = 9:1),

Rf = 0.79 (chloroform:methanol = 4:1).

EXAMPLE 2

0.5 g (0.57 mmol) 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine benzhydryl ester (cx,e mixture) containing 1.33 mol of water are hydrated in 20 ml of methanol-tetrahydrofuran (1:1) with 0.1 g of 5% palladium carbon as catalyst at normal pressure and room temperature. After one hour, the catalyst is filtered off and the filtrate is evaporated in vacuo at 30°C. The residue is dissolved in 10 ml of methanol-chloroform (1:1) and the crude product is precipitated from the solution obtained with diethyl ether. The precipitated crystals are filtered off and washed with diethyl ether.

The product obtained is dissolved in 50 ml of double-distilled water, filtered

twice through a millipore filter ("Nalgene S", 0.45 and 0.2 µm) and lyophilized. 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine-L-alanine (cx, p mixture) is obtained as a colorless powder containing 1.92 mol of water;

[cx]<g>° = + 19.5 ± 2.0'(c = 0.492; water),

Rf = 0.16 (chloroform:methanol:water = 70:30:5)

The starting material is produced in the following way

Step 2. 1

Analogously to example 1, one obtains from 2.11 g (4.07 mmol) N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine, which contains 1.56 mol of water with a total of 2.14 g (11, 06 mmol) diphenyldiazomethane in 42 ml methanol-dimethoxymethane (1:1) (18 hours, room temperature) N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine benzhydryl ester as colorless powder containing 2.24 mol water .

C35H47<N>5012-2.24H20 (770.14)

Calculated: C 54.58 H 6.76 N 9.09 0 29.59 H20 5.25

Found: C 54.85 H 6.86 N 9.110 29.21 H20 5.25

[a]D° = + 2.9 ± 2° (c = 0.478; dimethylformamide),

Rf = 0.57 (chloroform:methanol = 7:3),

Rf = 0.65 (chloroform-methanol:water = 70:30:5).

Step 2. 1

Analogously to example 1, one obtains from 1.19 g (1.54 mmol) N-propionyl-des-methylmur amyl-L-alanyl-D-isoglutaminyl-L-alanine benzhydryl ester (cx.e mixture) containing 2.24 mol water, with 0.7 ml (7.4 mmol) acetic anhydride in 12 ml pyridine (20 hours, room temperature) 1,4,6-tri-O-acetyl-N -propionyl-desmethylmuramyl-L -alanyl- D-isoglutaminyl-L-alanine benzhydryl ester (a,& mixture) as colorless powder containing 1.33 mol of water.

C41H53N5015-1,33H20 (879.85)

Calculated: C 55.97 H 6.40 N 7.96 O 29.69 H20 2.72

Found: C 56.14 H 6.36 N 7.98 = 29.56 H20 2.72

[cx]D<0>= + 23.8 ± 2.1° (c = 0.478; dimethylformamide),

Rf = 0.36 (chloroform:methanol = 9:1),

Rf = 0.69 (chloroform:methanol = 4:1),

Rf = 0.87 (chloroform:methanol = 7:3).

EXAMPLE 3

Analogous to example 2, one obtains by hydration of 0.5 g (0.64 mmol) 1,4,6-tri-O-acetyl-N-propionyl-des methylmur amyl-L-alanyl-D-isoglutamic acid-( ca) - n-butyl ester-(Cy)-benzhydryl ester (mainly α-anomers) containing 0.73 mol of water, in the presence of 0.1 g of 5% palladium carbon in 20 ml of methanol-tetrahydrofuran (1:1) (la,e), 4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid-(C0()-n-butyl ester as colorless crystals with m.p. 94-96" C (decomposition, from chloroform-diethyl ether [1:6]), which contains 0.46 moles of water;

[a] 5° = + 45.6 ±1.8° (c = 0.543; methanol),

Rf = 0.39 (chloroform:methanol =4:1),

Rf = 0.53 (chloroform:methanol = 7:3),

Rf = 0.59 (chloroform:methanol:water = 70:30:5).

The starting material is obtained in the following way:

Step 3. 1

Analogous to example 1, one obtains from 1.9 g (3.3 mmol) N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamic acid-(C0()-n-butyl ester), which contains 1.03 mol of water, with a total of 2 .5 g (12.85 mmol) diphenyldiazomethane in 40 ml methanol-dimethoxyethane (1:1; 70 hours, room temperature) N-propionyl-des methyl mur amy 1- L-a lanyl-D -iso glutamic acid - (Ca ) - n-buty lester-()-benzhydryl ester as colorless powder containing 0.99 mol of water.

C36H49N3O12-0.99H2O (733.62)

Calculated: C 58.94 H 7.03 N 5.73 O 28.32 H20 2.43

Found: C 59.24 H 7.06 N 5.59 O 28.08 H20 2.43

[cx]D° = + 12.5 ± 1.9°(c = 0.522; methanol)

Rf = 0.46 (chloroform:methanol = 9:1),

Rf = 0.63 (chloroform:methanol = 4:1),

Rf = 0.88 (chloroform:methanol = 7:3).

Step 3. 2

Analogous to example 1, one obtains from 1.15 g (1.57 mmol) N-propionyl-des-methylmur amyl-L-alany 1-D-isoglutamic acid re-(Ca)-n-butyl ester-(C^ )- benzhydryl ester still containing 0.99 mol of water, with 0.70 ml (7.4 mmol) of acetic anhydride in 12 ml of pyridine (16 h, room temperature) 1,4,6-tri- O -ac ethyl -N-propionyl -des methy lmur amyl -L-alanyl-D-isoglutamins yre-(Ccx)-n-butyl ester-(C^)-benzhydryl ester (cx,g mixture) as colorless powder containing 0.73 mol of water.

C42H55N3°15"0>73H2° (855.08)

Calculated: C 58.99 H 6.60 N 4.91 0 29.50 H20 1.54

Found: C 58.96 H 6.72 N 4.76 0 29.49 H20 1.54

[a]6° = + 12.6 ± 2.3° (c = 0.443; chloroform),

Rf = 0.72 (chloroform:methanol = 9:1),

Rf = 0.96 (chloroform:methanol = 4:1).

EXAMPLE 4

4.00 g (5.28 mmol) N-acetyl-(1a,p),4,6-tri-0-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester, dissolved in 40 ml methanol-water ( 3:1) is hydrogenated analogously to example 1 in the presence of 0.8 g of palladium-on-carbon (10%). The catalyst is filtered off, the filtrate is evaporated to dryness and the residue is chromatographed on silica gel ("type 60", Merck; 0.063-0.2 mm) (1:50) in the system chloroform-methanol-water (70:30:5) (fractions 1- 30; 20 ml; from 31: 5 ml). The material contained in fractions 3-45 is collected, dissolved in 50 ml of double-distilled water and lyophilized after filtration through a millipore filter (0.2 µm). N-acetyl-1,4,6-tri-0-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine (a,p mixture) is obtained as a colorless powder, which contains 1.31 mol of water.

C24H37N4°14'1'31H2° (629.17)

Calculated: C 45.82 H 6.38 N 8.91 H20 3.74

Found: C 45.9 H 6.6 N 9.1 H 2 O 3.7

[a]6° = +42 ± 1°(c = 0.768; methanol),

Rf = 0.65 (n-butanol:pyridine:acetic acid:water = 38:24:8:30),

Rf = 0.56 (ethyl acetate: n-butanol: pyridine: acetic acid: water

= 42:21:21:6:10),

Rf = 0.39 (ethyl acetate:acetic acid:water:methanol

= 67:10:23:12).

The starting material is produced in the following way:

Step 4. 1

A solution of 22.8 g (40 mmol) of N-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine in 500 ml of a 1:1 mixture of 1,2-dimethoxyethane and methanol is mixed with 11.6 g (60 mmol) diphenyldiazomethane, and the solution is stirred for 16 hours at room temperature. The red suspension is evaporated at 20°C under reduced pressure, and the residue is triturated several times with diethyl ether, until an almost colorless product is obtained. This is dissolved in 100 ml of methanol and brought to crystallization by portionwise addition of a 2:1 mixture of diethylene petroleum ether. After stirring for several hours, first at room temperature, then in an ice bath, the crystal mass is filtered off and dried under reduced pressure. N-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester is obtained in the form of cube-shaped crystals. Temp. 170 °C (with decomposition).

[cx]D° = + 14 ± 1°(c = 1.5, methanol)

Rf = 0.40 (chloroform:methanol:water = 70:30:5)

Rf = 0.66 (ethyl acetate: n-butanol: pyridine: glacial acetic acid: water

= 42:21:21:6:10).

Step 4. 2

Analogous to step 1.2, 3.69 g (5.6 mmol) of N-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester, dissolved in 40 ml of abs. pyridine, with 2.04 g (20 mmol) of acetic anhydride (1 hour, room temperature). The clear solution is evaporated vigorously at 30 "C, the residue is taken up in 150 ml of ethyl acetic acid and the solution is extracted twice with 50 ml of water. After drying and evaporation of the solvent, N-acetyl-(lex,3),4,6-tr i-O is obtained - acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester.

[cx]D° = 33<»>± 1- (c = 1.159, methanol),

Rf = 0.70 (chloroform:methanol:water 70:30:5),

Rf = 0.48 (n-butanol:acetic acid:water = 75:7.5:21) and

Rf = 0.81 (ethyl acetate: n-butanol: pyridine: acetic acid: water

= 42:21:6:10).

EXAMPLE 5

Analogous to example 4, 0.60 g (0.76 mmol) of N-acetyl-1,4,6-tri-0-acetyl-desmethylmuramyl-L-α-aminobutyryl-D-isoglutamine-benzhydryl ester, dissolved in 20 ml of dimethoxyethane, is hydrogenated water (20:1) in the presence of 0.2 g palladium-on-carbon (10%). The residue is purified by chromatography on silica gel in the system chloroform-methanol-water (70:30:5). The fractions containing the material are collected, dissolved in 10 ml of dimethoxyethane-water (1:1) and filtered to partially remove the Na salt formed over a strongly acidic ion exchanger ("Dowex 50", X8/particle diameter 0.149-0.074 mm; 50 ml) and washed with a total of 150 ml of mixture. The combined filtrates are evaporated on a rotary evaporator at approx. 30°C, the residue is dissolved in 10 ml of a mixture of tert.-butanol-water (2:1) and lyophilized after filtration through a millipore filter ("Fluoropore", PTFE, 0.2 pm). N-acetyl-1,4,6-tri-0-acetyl-desmethylmuramyl-L-α-aminobutyryl-D-isoglutamine (cx,e mixture) is obtained as a colorless powder containing 1.94 mol of water.

C25<H>38<N>4014'1.94 mol H20 (653.54)

Calculated: C 45.95 H 6.49 N 8.57 H20 5.34

Found: C 46.2 H 6.6 N 8.3 H 2 O 5.3

[cx]D° = + 34 ± 1* (c = 0.511; chloroform),

Rf = 0.15 (chloroform:methanol:water = 70:30:5),

Rf = 0.58 (ethyl acetate: n-butanol: pyridine: acetic acid: water

= 42:21:21:6:10).

The starting material is obtained in the following way:

Step 5. 1

Analogously to step 4.2, one obtains from 0.55 g (0.85 mmol) N-acetyl-desmethylmuramyl-L-cx-aminobutyryl-D-isoglutamine-benzhydryl ester, dissolved in 5 ml abs. pyridine, and 0.281 g (2.76 mmol) acetic anhydride (30 minutes,

room temperature) N-acetyl-(lcx,3),4,6-tri-O-acetyl-desmethylmuramyl-L-cx-aminobutyryl-D-isoglutamine benzhydryl ester.

Rf = 0.63 (chloroform:methanol:water = 70:30:5) and

Rf = 0.82 (ethyl acetate: n-butanol: pyridine: acetic acid: water

= 42:21:21:6:10).

in

EXAMPLE 6

Analogous to example 5, hydrogenolysis of 1.02 g (1 mmol) of N-acetyl-1,4,6-0-acetyl-desmethylmuramyl-L-cx-amino butyryl-D-glutamic acid dibenzhydryl ester is obtained, then chromatography on silica gel and treatment with "Dowex 50" (H form) N-acetyl-1,4,6-tri-0-acetyl-desmethylmuramyl-L-cx-aminobutyryl-D-glutamic acid (cx, 3 mixture) as colorless, loose powder containing 1.38 moles of water.

<C>25<H>37N3<0>15'lm38H20 (644.44)

Calculated: C 46.59 H 6.26 N 6.52 H20 3.87

Found: C 46.8 H 5.9 N 6.5 H 2 O 3.9

[a]D° = +16 ± 1°(c = 0.666; chloroform),

Rf = 0.23 (chloroform:methanol:water = 70:50:5),

Rf = 0.43 (n-butanol:pyridine:acetic acid:water = 38:24:8:30),

Rf = 0.32 (ethyl acetate: n-butanol: pyridine: acetic acid: water

= 42:21:21:6:10).

The starting material is produced in the following way:

Step 6. 1

3.00 g (6.1 mmol) of N-acetyl-desmethylmuramyl-L-cx-aminobutyryl-D-glutamic acid, dissolved in 30 ml of a 1:1 mixture of dimethylformamide and methanol, is mixed at room temperature and under stirring with a excess of diphenyldiazomethane. After 3 days, the red suspension is evaporated under high vacuum at 30 °C, the red residue is triturated several times with diethyl ether-petroleum ether (1:3) and the supernatant liquid is decanted. The solid residue is first purified by chromatography on 600 g of silica gel ("Type 60", Merck) in the system chloroform-methanol-water (70:30:5; among other things to remove half-esters), then on a second column (100 g) first with ethyl acetate, then ethyl acetate-methanol (3:1; 5 ml fractions).

The combined material is dissolved in 4.5 ml of methanol and lyophilized after adding 70 ml of abs. dioxane and regular filtration through a millipore filter. N-acetyl-desmethylmuramyl-L-a-aminobutyryl-D-glutamic acid dibenzhydryl ester is obtained as a colorless powder containing 0.93 mol of water.

C45H51N3°12"0'93H2° (842.67)

Calculated: C 64.14 H 6.34 N 4.99 H20 1.99

Found: C 64.1 H 6.5 N 4.9 H 2 O 2.0

[a]D° = + 11.6 ± 2.9"(c = 0.344; methanol),

Rf = 0.55 (acetic acid ethyl ester methanol = 4:1) and

Rf = 0.77 (chloroform-isopropanol = 1:1).

Step 6. 2

From 2.10 g (2.54 mmol) of N-acetyl-desmethylmuramyl-L-cx-aminobutyryl-D-glutamic acid dibenzhydryl ester and 1.053 ml (11.43 mmol) of acetic anhydride in 6 ml of absolute pyridine, one obtains analogously to example 1 after 12 hour's rest at room temperature the peracetyl compound. The purification takes place on 450 g of silica gel ("Type 60", Merck; grain size 0.000063-0.200 mm; 5 ml fractions) first with chloroform, then from fraction 20 with chloroform:isopropanol (3:1). The material contained in fractions 43-106 is collected, dissolved in 5 ml of chloroform, filtered by adding 40 ml of abs. dioxane through a millipore filter ("Fluoropore", PTFE, 0.2 pm) and then lyophilized. N-acetyl-(lcx,p),4,6-tri-0-acetyl-desmethylmuramyl-L-cx-aminobutyryl-D-glutamic acid dibenzhydryl ester is obtained as a colorless, loose powder containing 0.37 mol of water.

C5l<H>57N3Oi5'0.37H2O (958.69)

Calculated: C 63.90 H 6.09 N 4.38 H20 0.69

Found: C 63.6 H 6.1 N 4.6 H 2 O 0.70

[a]5° = + 29.6 ± 1.7° (c = 0.577; methanol),

Rf = 0.69 (n-butanol:acetic acid:water = 75:7.5:21) and

Rf = 0.93 (ethyl acetate: n-butanol: pyridine: acetic acid: water

= 42:21:21:6:10).

EXAMPLE 7

Analogously to example 5, hydrogenolysis of 0.80 g (1 mmol) of N-acetyl-1,4,6-tri-O-acety 1-desmethylmuramy 1-L-valy 1-D-isoglutamin-benzhydryl ester is obtained, dissolved in 20 ml of dimethoxyethane-water (4:1) and ordinary chromatography N-acetyl-1,4,5-tri-0-acetyl-desmethylmuramyl-L-valyl-D-isoglutamine (cx,<p->mixture) as colorless lyophilisate containing 0.9 mol of water.

[cx]D° = +17 ± 1* (c = 0.352; chloroform),

Rf = 0.19 (chloroform:methanol:water = 70:30:5),

Rf a0 0.28 (n-butanol:acetic acid:water = 75:7.5:21),

Rf = 0.63 (ethyl acetate: n-butanol: pyridine: acetic acid: water

= 42:21:21:6:10).

The starting material is produced in the following way:

Step 7. 1

1.60 g (3.15 mmol) of N-acetyl-desmethylmuramyl-L-valyl-D-isoglutamine, dissolved in 30 ml of methanol, esterified with an excess of diphenyldiazomethane (2 hours, room temperature). The product is purified by precipitation several times from methanol-diethyl ether (1:8). N-acetyl-desmethylmuramyl-L-valyl-D-isoglutamine benzhydryl ester is obtained.

[a]D°= + 13°(c = 1.067; methanol),

Rf = 0.56 (chloroform:methanol:water = 70:30:5),

Rf = 0.42 (n-butanol:acetic acid:water = 75:7.5:21) and

Rf = 0.72 (ethyl acetate: n-butanol: pyridine: acetic acid: water

= 42:21:21:6:10).

Step 7. 2

Analogously to step 1.2, one obtains from 0.67 g (1 mmol) N-acetyl-desmethylmura-methyl-L-valyl-D-isoglutamine-benzhydryl ester and 0.367 g (3.6 mmol) acetic anhydride in 5 ml abs. pyridine (30 minutes, room temperature) N-acetyl-(1a,3),4,6-tri-O-acetyl-desmethylmuramyl-L-valyl-D-isoglutamine-benzhydryl ester.

[a]j)° = + 35°P 1°(c = 0.594; methanol),

Rf = 0.92 (chloroform:methanol:water = 70:30:5) and

Rf = 0.80 (ethyl acetate: n-butanol: pyridine: acetic acid: water

=42:21:21:6:10).

EXAMPLE 8

Analogously to example 5, hydrogenolysis of 0.86 g (0.87 mmol) of N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-valyl 1-D-glutamic acid e-dibenshydryl ester and ordinary chromatography N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-valyl-D-glutamic acid (α,3 mixture) as colorless lyophilisate containing 1.15 moles of water;

C27H4iN3015-1,15H20 (688.35)

i Calculated: C 48.52 H 6.56 N 6.29 H20 3.11

Found: C 48.9 H 6.5 N 6.7 H 2 O 3.1

[a]D° = +44 ± 1°(c = 0.429; water),

Rf = 0.08 (chloroform:methanol:water = 70:30:5),

Rf = 0.48 (n-butanol:pyridine:acetic acid:water = 38:24:8:30),

Rf = 0.48 (ethyl acetate:acetic acid:water:methanol = 67:10:23:12).

The starting material is produced in the following way:

Step 8. 1

1.80 g (3.34 mmol) of N-acetyl-muramyl-L-valyl-D-glutamic acid, dissolved in 30 ml of methanol, are mixed while stirring with an excess of diphenyldiazomethane. After stirring for 3 hours at room temperature, the reaction solution is evaporated and the red, oily residue is decolored by trituration several times with diethyl ether-petroleum ether (1:1). The semi-solid material is taken up in 20 ml of dimethoxyethane and precipitated with the dropwise addition of a total of 100 ml of petroleum ether. N-acetyl-muramyl-L-valyl-D-glutamic acid dibenzhydryl ester is obtained as a colorless, amorphous powder.

[cx]D° = + 33 ± 1° (c = 0.489; methanol),

Rf = 0.62 (chloroform:methanol:water = 70:30:5),

Rf = 0.66 (n-butanol:acetic acid:water = 75:7.5:21).

Step 8. 2

0.845 g (1 mmol) N-acetyl-muramyl-L-valyl-D-glutamic acid dibenzhydryl ester, dissolved in 10 ml abs. pyridine is mixed with stirring at 0°C with 0.367 g (3.6 mmol) of acetic anhydride and allowed to stand for 45 minutes. 1 ml of distilled water is added and evaporated at 25 °C to dryness. The residue is taken up in ethyl acetate, the solution is washed several times with water, dried over sodium sulphate and evaporated. N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-valyl-D-glutamic acid dibenzhydryl ester is obtained as an amorphous powder.

in

Rf = 0.76 (n-butanol:pyridine:acetic acid:water = 38:24:8:30),

Rf = 0.91 (ethyl acetate: n-butanol: pyridine: acetic acid: water

= 42:21:21:6:10).

; EXAMPLE 9

Analogously, the following compounds are obtained: N-acetyl-1,4,6-tri-0-propionyl-muramyl-L-alanyl-D-isoglutaminyl-L-alanine, N-acetyl-1,4,6-tri-0- propionyl-muramyl-L-alanyl-D-isoglutaminyl-L-lysine, N-acetyl-l,4,6-tri-0-acetyl-muramyl-L-N-methyl-alanyl-D-isoglutamine, N-acetyl-l, 4,6-tri-0-acetyl-muramyl-L-a-methyl-alanyl-D-isoglutamine, N-acetyl-l,4,6-tri-0-acetyl-muramyl-L-prolyl-D-isoglutamine, N- acetyl -1,4,6 -tri- O -acetyl -de smethylmu ramy 1-L- alanyl-D --y-carboxy-isoglutamine-methyl ester and N-ac ethyl -1,4,6- tri-O-actyl- N-benzoyl-desmethylmuramyl-L-cx-aminobutyryl-D-isoglutamine.

EXAMPLE 10

1.45 g (2.53 mmol) N-benzoyl-desmethylmuramyl-L-alanyl-D-glutamic acid dimethyl ester in 15 ml pyridine is mixed with 0.1 g 4-dimethylaminopyridine and 1.5 g (1.24 ml) benzoyl chloride . After 20 hours at room temperature, it is heated for 15 minutes to 40 °C and, after cooling, 10 ml of water is added. The solution is then evaporated in vacuo to a syrup, taken up with chloroform and extracted successively with 1N hydrochloric acid, 5% NaHCO 3 solution and water. After drying (Na2SO4) and evaporation of the organic phase, a mixture of (lcx,g)4,6-tri-0-benzoyl-N-benzoyl-desmethylmuramyl-L-alanyl-D-glutamic acid dimethyl ester and (lcx,< p>),6-di - O -benzoyl -N-b enzoyl -d esmethylm uramyl -L -alanyl - D -glutam insy r edimethyl - ester.

By chromatography on 150 g of silica gel (Merck, 0.04-0.063 mm) with CH2CI2-acetic acid ethyl ester (1:1; fractions 1-4, in each case 500 ml eluent) CH2CI2-acetic acid ethyl ester (3:7; fractions 5-8, in each case 500 ml of eluent) the mixture can be divided. The four compounds obtained show the following Rf values (in all cases in chloroform:acetone = 6:4): 0.82 (1a,4,6-tri-0-benzoyl compound), 0.72 g (l<p >,4,6-tri-O-benzoyl compound), 0.55 (lcx,6-di-O-benzoyl compound) and 0.36 (l<p>,6-di-O-benzoyl compound ), the [cx]^ values amount in the order mentioned: +40.6°, -24.8°, +33.8° and -90.1° (all in chloroform). The melting points are in the order mentioned: 92-96 °C, 177-178 °C, 151-152 °C and 134-136 °C.

EXAMPLE 11

Analogous to example 1, by hydrogenolysis of N-acetyl-1,4,6-tri-0-acetyl-muramyl-L-valyl-D-isoglutamine benzhydrylase (cx, 3-mixture) N-acetyl-

(1a,p),4,6-tri-O-acetyl-muramyl-L-valyl-D-isoglutamine-1,6 H2O as colorless powder;

[a]D° = 56 ± 1- (c = 0.943; water),

Rf = 0.4 (dichloromethane: methanol: glacial acetic acid = 20:5:1).

The starting material is obtained in the following way:

Step 11. 1

A solution of 2.0 g of N-acetyl-muramyl-L-valyl-D-isoglutamine in 25 ml of methanol and 25 ml of 1,2-dimethoxyethane is mixed with 1.1 g of diphenyldiazomethane and stirred for 20 hours at room temperature. The reaction mixture is evaporated to dryness and extracted with diethyl ether. The residue is suspended in water, stirred, filtered off and dried over NaOH pellets. The obtained N-acetyl-muramyl-D-isoglutamine-benzhydryl ester melts at 185 °C (decomposition),

Rf = 0.5 (chloroform:methanol:water = 14:6:1),

[α]D° = +38° (c = 0.912; methanol).

Step 11. 2

A solution of 1.2 g of N-acetyl-muramyl-L-valyl-D-isoglutamine-benzhydryl ester in 12 ml of absolute pyridine is mixed with stirring with 0.75 ml of acetic anhydride and allowed to stand for 24 hours at room temperature. The reaction mixture is then poured into 40 ml of ice water, whereby the product crystallizes out. The crystals are suctioned off, washed with water and dried. After recrystallization from acetic acid ethyl ester and diethyl ether, N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-valyl-D-isoglutamine-benzhydryl ester is obtained,

Rf = 0.3 (methylene chloride: methanol = 5:1).

EXAMPLE 12

Analogous to example 1, by hydrogenolysis of N-acetyl-1,4,6-tri-0-acetyl-muramyl-L-alanyl-D-isoglutamine-benzhydryl ester (a, 3-mixture) N-acetyl-(lc<, e), 4,6-tri-O-acetyl-muramyl-L-alanyl-D-isoglutamine-1,8 as colorless lyophilisate;

[a]D° = +57, ±1°(c = 0.923@; water),

Rf = 0.4 (dichloromethane: methanol: acetic acid = 20:5:1).

The starting material is produced in the following way:

Step 12. 1 A solution of 1.1 g of N-acetyl-muramyl-L-alanyl-D-isoglutamine in 12.5 ml of methanol and 12.5 ml of 1,2-dimethoxyethane is mixed with 0.42 g of diphenyldiazomethane and stirred in 20 hours at 40 "C. After evaporation of the solvent, the residue is extracted thoroughly with diethyl ether and water and dried over NaOH pellets. N-acetyl-muramyl-L-alanyl-D-isoglutamine-benzhydryl ester is obtained in the form of white crystals.

Rf = 0.6 (methylene chloride:methanol:water = 14:6:1).

Step 12. 2

A solution of 2.0 g of N-acetyl-muramyl-L-alanyl-D-isoglutamine-benzhydryl ester in 20 ml of absolute pyridine is mixed with stirring with 1.7 ml of acetic anhydride and allowed to stand for 24 hours at room temperature. The reaction mixture is then poured onto 50 ml of ice water. The precipitated material is taken up in 100 ml of acetic acid ethyl ester, washed with dilute hydrochloric acid and half-saturated sodium chloride solution and dried over magnesium sulfate. After evaporation of the solvent, N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-alanyl-D-isoglutamine benzhydryl ester is obtained.

Rf = 0.7 (methylene chloride: methanol = 5:1).

EXAMPLE 13

In an analogous way, the following compounds are obtained: N-b enzoyl lam ino -1,4,6-t ri-0 -n-h exa noyl -des methyl lmur amyl -L-a lany 1-D-isoglutamine [Rf = 0.67 (chloroform:methanol = 17:3)], N-propionyl-1,4,6-tri-0-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine, N-acetyl-1,4,6-tri-0-propionyl-muramyl -L-α-aminobutyryl-D-isoglutamine, N-ac ethyl -1,4,6-tri-O-acetyl-desmethylmuramyl-L-al anyl-D-isoglutam inic acid diamide,

1,4,6-tri-O-acetyl-N-benzoyl-desmethylmur amyl-L-alanyl-D-glutamic acid dimethyl ester,

N-Acetyl -1,4,6-tri-O-acetyl-desmethylmuramyl-L-alanyl-D-glyutamins ure-dimethyl ester,

N-ac ethyl -1,4,6-tri-0-acetyl - L-a -aminobutyryl-D-isoglutaminyl-L-arginine methyl ester and

N-Acethyl-1,4,6-tri-O-acetyl-muramyl-L-alanyl-D-isoglutaminyl-N-benzyloxycarbonyl-4-thialysine-isopropyl ester sulfoxide.

EXAMPLE 14

Female MF-2f SPF mice with a body weight of 14-16 g are infected under light anesthesia intranasally with a mixture of equal parts of diethyl ether, ethanol and chloroform with lethal doses (approx. 1 LD§q_9q; 1-4 "plaque forming units"

[PFU]) of in each case 0.05 ml of a suspension of influenza A/Texas/1/77 viruses (mouse-adapted strain).

Each mouse from groups of 10 of these mice is administered at the time indicated below [days] counted from the day of infection once (single dose) the quantity of the relevant active substance indicated in table 1 in 0.05 or in 0.2 ml of a 0.005% by weight solution of kårbok sym ethyl cellulose sodium salt in double distilled, pyrogen-free water in the case of intranasal resp. oral administration of the type indicated in Table 1. 20 of the above-mentioned infected mice for control, i.e. they receive a placebo (0.005% by weight solution of carboxymethylcellulose sodium salt).

The intranasal delivery of the active substance is carried out under light anesthesia with a mixture of equal parts of diethyl ether, ethanol and chloroform.

Compound I = 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine.

EXAMPLE 15: Non-aqueous single dose for nasal delivery Composition:

(1a,g),4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamic acid-(Ca)-n-butyl-

0.03 mg of the active substance is dissolved under aseptic conditions in 29.97 mg of "Miglyol".

This solution is filled in a commercially available disposable nasal applicator, to which a dose of fuel is added before use.

EXAMPLE 16: Nasal drops

Composition:

Sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium chloride, thiomersal and EDTA disodium salt are dissolved in part of the above-mentioned amount of demineralized water with stirring at room temperature.

The active substance and the rest of the demineralized water are then dissolved in this solution.

The solution or a part of it or a multiple thereof is filtered using a membrane filter and filled in suitable containers. Suitable containers are e.g. a) Glass or plastic containers (a 5 ml or 10 ml), which have a glass or plastic pipette with an elastomer pipette aspirator, b) Collapsible plastic bottles with a riser and a plastic syringe head,

c) Single dose containers made of plastic (content 2-3 drops) or

d) Glass or plastic bottles that are equipped with a standard

pump dosing spray made of plastic (without propellant gas).

EXAMPLE 17: Nasal ointment

Composition:

(la,p),4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid-(Ca)-n-butyl-

The fat phase, consisting of paraffin oil, vaseline and wool grease, is melted together. The aqueous solution of the active substance is incorporated into the fat phase at approx. 50 °C.

EXAMPLE 18: Preparation of 1000 tablets containing

0.5% active substance

Composition per 1000 tablets: (lcx,e),4,6-tri-0-acetyl-N-propionyi-desmethylmuramyl-L-alanyl-D-glutamic acid-(C0()-n-butyl-

The active substance and 15 g of lactose are mixed together. The obtained mixture is mixed together with 28 g of lactose and 47 g of corn starch. With the obtained mixture and an aqueous solution of "Pharmcoat" a mass ready for granulation is produced which is granulated, dried and ground. To this, 5 g of corn starch, "Aerosil" and magnesium stearate are mixed and compressed into 1000 tablets, each weighing 100 mg.

The pressed tablets can be varnished in a manner known per se with gastric juice-resistant varnish.

EXAMPLE 19

0.070 g (0.088 mml) N-acetyl-1,4,6-tri-0-acetyl-muramyl-L-N-methyl-alanyl-D-isoglutamine-benzhydryl ester (cx, p-mixture) is hydrated analogously to example 5, but in glacial acetic acid . The solution is lyophilized after removal of the catalyst. The residue is taken up in 3 ml of chloroform, poured through a millipore filter (PTFE; 0.2 pm) and the solution is lyophilized after adding 60 ml of abs. dioxane. N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-N-methyl-alanyl-D-isoglutamine (cx, p mixture) is obtained as a colorless powder;

[cx]D° = +34.9 ± 1.1°(c = 0.946; methanol)

Rf = 0.25 (chloroform:methanol:water = 70:30:5),

Rf = 0.43 (acetonitrile water = 3:1),

Rf = 0.21 (n-butanol:acetic acid:water = 75:7, 5:21).

The starting material is produced in the following way:

Step 19. 1

0.50 g (0.99 mmol) N-acetyl-muramyl-L-N-methyl-alanyl-D-isoglutamine (a, p-mixture) is transferred analogously to step 8.1 to N-acetyl-muramyl-L-N-methyl-alanyl-D -isoglutamine benzhydryl ester.

Rf = 0.76 (acetonitrile water = 3:1)

Rf = 0.63 (chloroform:methanol:water = 70:30:5).

Step 19. 2

The crude product obtained according to step 19.1 is acetylated analogously to step 8.2 and purified by chromatography on silica gel (1:180) in chloroform-isopropanol (9:1;

0.8 ml fractions). The uniform fractions are lyophilized from abs. dioxane. N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-N-methyl-alanyl-D-isoglutamine-benzhydryl ester (ot,3 mixture) is obtained as a colorless powder;

Rf = 0.61 (n-butanol:acetic acid:water = 75:7.5:21),

Rf = 0.20 (chloroform: isopropanol: glacial acetic acid = 70:8:2).

EXAMPLE 20

0.32 g (0.40 mmol) N-acetyl-1,4,6-tri-0-acetyl-α-amino-isobutyryl-D-isoglutamine-benzhydryl ester (α,3 mixture), dissolved in 8 ml of glacial acetic acid is hydrated analogously to example 5, but in glacial acetic acid. The catalyst is filtered off and the solution is filtered through a millipore filter (PTFE; 0.2 µm). After lyophilization, N-acetyl-1,4,6-tri-0-acetyl-muramyl-cx-amino-isobutyryl-D-isoglutamine (a,3 mixture) is obtained as a colorless powder containing 0.81 mol of water;

[a]&° = +55.8 ± 1.1°(c = 0.923; methanol),

Rf = 0.40 (chloroform:methanol:water = 70:30:5),

Rf = 0.70 (ethyl acetate:pyridine:acetic acid:water = 62:21:6:11),

Rf = 0.73 (ethyl acetate: n-butanol: pyridine: acetic acid: water

= 42:21:21:6:10).

The starting material is produced in the following way:

Step 20. 1

0.50 g (0.99 mmol) of N-acetyl-α-amino-isobutyryl-D-isoglutamine (cx, 3 mixture) is esterified analogously to step 8.1 with diphenyldiazomethane. The crude product [Rf = 0.48 (chloroform:methanol:water = 70:30:5)] is peracylated analogously to step 8.2. The reaction solution is evaporated in vacuo to dryness, taken up in water

and lyophilized. The purification takes place on silica gel (1:100) in chloroform-methanol

in (9:1; 1 ml fractions). The pure fractions are collected, taken up in 5 ml of chloroform, filtered (PTFE; 0.2 µm) and then evaporated to dryness. The rest is recorded in abs. dioxane and lyophilized. N-acetyl-1,4,6-tri-0-acetyl-muramyl-cx-amino-isobutyryl-D-isoglutamine-benzhydryl ester (a,3 mixture) is obtained

as colorless powder containing 0.73 mol of water.

in

[cx]D0 = +45.7 ±1°(c = 0.993; methanol),

Rf = 0.80 (chloroform:methanol:water = 70:30:5),

Rf = 0.32 (chloroform-methanol = 9:1),

Rf = 0.50 (n-butanol:acetic acid:water = 75:7.5:21).

EXAMPLE 21

0.30 g (0.37 mmol) of N-acetyl-1,4,6-tri-0-acetyl-muramyl-L-prolyl-D-isoglutamine-benzhydryl ester (cx, p-mixture) is hydrated analogously to example 20 in glacial acetic acid . After filtration (PTFE; 0.2 µm) and lyophilization, N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-prolyl-D-isoglutamine (cx, p mixture) is obtained as a colorless powder;

[cx]j}0 = +51.4 ±1°(c = 0.956; methanol),

Rf = 0.20 (chloroform:methanol:water = 70:30:5),

Rf = 0.39 (acetonitrile:water = 3:1).

The starting material is produced in the following way:

Step 21. 1

0.50 g (0.934 mmol) of N-acetyl-muramyl-L-prolyl-D-isoglutamine (cx,P mixture) is esterified analogously to step 8.1 with diphenyldiazomethane. After usual work-up, N-acetyl-muramyl-L-prolyl-D-isoglutamine-benzhydryl ester (cx,<p->mixture) is obtained as a colorless powder;

[cx]D° = +30.4 ±1.6° (c = 0.608; methanol),

Rf = 0.63 (chloroform:methanol:water = 70:30:5).

in

Step 21. 2

N-acetyl-muramyl-L-proly 1-D-isoglutamine-benzhydryl ester is peracetylated analogously to step 8.2. After lyophilization from abs. dioxane gives N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-propyl-D-isoglutamine-benzhydryl ester r-hydrate in (a,<p->mixture) as colorless powder;

[cx]D° = +32.2 ±1.6° (c = 0.645; dimethylsulfoxide),

Rf = 0.92 (chloroform:methanol:water = 70:30:5),

Rf = 0.60 (n-butanol:acetic acid:water = 75:7.5:21).

in

EXAMPLE 22

N-acetyl -1,4,6-tri-O-acet<y>l-desmet<y>lmuram<y>l-L-alan<y>l-D-7-benzhydryloxycarbonyl-isoglutamine methyl ester (cx,3-mixture ) is hydrated analogously to example 5, but in glacial acetic acid. After usual work-up, filtration (PTFE; 0.2 pm) and lyophilization, N-acetyl-1,4,6-tri-0-acetyl-desmethylmuramyl-L-alanyl-D-7-carboxy-isoglutamine methyl ester (cx ,3 mixture) as colorless powder;

Rf = 0.66 (chloroform:methanol:water:acetic acid = 55:47:13:5),

Rf = 0.18 (chloroform:methanol:water = 70:30:5).

The starting material is produced in the following way:

Step 22. 1

0.216 g (0.40 mmol) of N-acetyl-desmethylmuramyl-L-alanyl-D--Y-carboxy-isoglutamine (a,3-mixture) is esterified analogously to step 8.1 with diphenyldiazomethane (3 equivalents). The bright red suspension is filtered after stirring for 3 hours and the filtrate is evaporated to dryness. The mixture, consisting of addition product, mono- and dibenzhydryl ester as well as decomposition products, originating from diphenyldiazomethane, can be separated by chromatography on silica gel (1:100) in the system chloroform:methanol:water =

(70:30:5). After two times of chromatography, N-acetyl-desmethylmuramyl-L-alanyl-D--y-benzhydryloxycarbonyl-isoglutamine-benzhydryl ester (a, 3-mixture) is obtained as a colorless resin with the Rf values

Rf = 0.67 (chloroform:methanol:water = 70:30:5) and

Rf = 0.46 (n-butanol:acetic acid:water = 75:7.5:21)<*>

and N-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester with the Rf value

Rf = 0.13 (chloroform:methanol:water = 70:30:5).

Step 22. 2

N- ac ety 1- de sm ethyl mu ra my 1- L- al an yl -D -i so gl ut amin -benzhydr yl st is esterified in methanolic solution as usual with a solution of diazomethane in diethyl ether. After evaporation, N-acetyl-desmethyl-mur amy 1-L-alan yl-D-y-b en zhy dry lox ykar bony 1-is oglu tami n-methyl ester (cx,3 mixture) is obtained as a colorless resin;

Rf = 0.13 (chloroform:methanol:water = 70:30:5).

Step 22. 3

Analogously to step 8.2, one obtains from N-acetyl-desmethylmuramyl-L-alanyl-D-Y-benzhydryloxycarbonyl-isoglutamine methyl ester (cx, p<->mixture), N-acetyl - 1,4,6-tr i-0 -acet yl -desmethylmuramyl-L-ala nyl-D-T-benzhydryloxycarbonyl-isoglutamine methyl ester (cx,p mixture) as lyophilisate;

Rf = 0.67 (chloroform:methanol:water = 70:30:5),

Rf = 0.35 (n-butanol:acetic acid:water = 75:7.5:21).

Example 23

Analogous to example 5, but in glacial acetic acid, one obtains from N-acetyl-1,4,6-tri-O-acetyl-desmethylm uramyl-L-alanyl-D--y-benz hydroxycarbonyl-isoglutamine-benzhydryl ester (a, p -mixture), N-acetyl-1,4,6-tri-O-acetyl-desmethylmuramyl-L-alanyl-D-^-carboxy-isoglutamine as colorless loose powder;

[cx]D° = +35.7 ±1.6°(c = 0.613; methanol),

Rf = 0.095 (chloroform:methanol:water = 70:30:5)

Rf = 0.093 (n-butanol:acetic acid:water = 75:7.5:21).

The starting material is produced in the following way:

Step 23. 1

48.5 mg (0.05 mmol) of N-acetyl-desmethylmuramyl-L-alanyl-D-7-benzhydryloxycarbonyl-isoglutamine-benzhydryl ester (a, p mixture) (see step 22.1) is peracetylated analogously to step 8.2. The reaction solution is diluted with a little abs. dimethylformamide and evaporated in HV to dryness. The obtained resin crystallizes after absorption in a little methanol (approx. 10% solution). N-acetyl-1,4,6-tri-O-acetyl-desmethylmuramyl-L-alanyl-D--Y-benzhydryloxycarbonyl-isoglutamine-benzhydryl ester (a,p mixture) is obtained in the form of colorless needles; m.p. 188-190°C (decomposition),

Rf = 0.85 (chloroform:methanol:water = 70:30:5),

Rf = 0.23 (chloroform: isopropanol: glacial acetic acid = 70:8:2),

Rf = 0.49 (n-butanol:acetic acid:water = 75:7.5:21).

EXAMPLE 24

0.200 g (0.236 mmol) 1,4,6-tri-O-acetyl-N-benzoyl-desmethylmuramyl-L-cx-aminobutyryl-D-isoglutamine-benzhydryl ester (a,3 mixture) is hydrated analogously to example 5, but in glacial acetic acid . After filtration (PTFE; 0.2 pm) and lyophilization from abs. dioxane gives 1,4,6-tri-O-acetyl-N-benzoyl-desmethylmuramyl-L-cx-aminobutyryl-D-isoglutamine (cx,3 mixture) as a colorless powder containing 0.75 mol of water.

[cx]D° = +33.9 ±1.1° (c = 0.885; chloroform),

Rf =0.44 (n-butanol:acetic acid:water = 75:7.5:21),

Rf = 0.60 (acetonitrile:water = 3:1).

The starting material is produced in the following way:

Step 24. 1

0.83 g (1.44 mmol) of N-benzoyl-desmethylmuramyl-L-α-amino-butyryl-D-isoglutamine sodium salt (cx, p-mixture) is esterified analogously to step 8.1 with diphenyldiazomethane in the presence of 1.3 mmol of trifluoroacetic acid (90 % of theoretical). The crude product is taken up in 20 ml of n-butanol, the solution is extracted three times, each time with 5 ml of water, and the aqueous phases are back-extracted twice, each time with 10 ml of n-butanol. The combined upper phases are dried over Na2S04 and evaporated to approx. 5 ml. The product is precipitated by adding 150 ml of diethyl ether. N-benzoyl-desmethylmuramyl-L-cx-aminobutyryl-D-isoglutamine-benzhydryl ester (cx, p-mixture) is obtained as a colorless powder;

[cx]J)° = +20.7 ±1°(c = 0.483; dimethylformamide),

Rf = 0.71 (chloroform:methanol:water = 70:30:5).

Step 24. 2

0.40 g (0.55 mmol) of N-benzoyl-desmethylmuramyl-L-cx-aminobutyryl-D-isoglutamine-benzhydryl ester (cx, p-mixture) is peracetylated analogously to step 8.2. The crude product is purified on silica gel (1:100) in the system chloroform-isopropanol (9:1; 0.8 ml fractions). The pure fractions are recorded in abs. dioxane and lyophilized after filtration (PTFE; 0.2 µm). 1,4,6-tri-O-acetyl-N-;benzoyl-desmethylmuramyl-L-cx-aminobutyry 1-D-isoglutamine- benzhydryl ester (cx,<p->mixture) is obtained as a colorless powder containing 0 .61 moles of water.

[c<]D<0>= +40.7 ± 1.1°(c = 0.910; methanol),

Rf = 0.38 (chloroform:methanol:water = 70:30:5),

Rf = 0.90 (acetonitric water = 3:1).

EXAMPLE 25

1.29 g (1.53 mmol) of N-acetyl-1,4,6-tri-0-propionyl-muramyl-L-cx-aminobutyryl-D-isoglutamine-benzhydryl ester (a, 3-mixture) are dissolved in glacial acetic acid and is hydrated analogously to example 8. The residue obtained after freeze-drying is taken up in 30 ml of water and extracted twice with 30 ml of diethyl ether. The water phase is freed of ether in vacuum, filtered ("Naglene"; 0.2 pm) and lyophilized. N-acetyl-1,4,6-tri-O-propionyl-muramyl-L-α-aminobutyryl-D-isoglutamine (α,<p->mixture) is obtained as colorless, loose powder containing 1.26 mol of water;

[cx]D<0>= +64.5 ± 0.9°(c = 1.106; methanol),

Rf = 0.53 (chloroform:methanol:water:acetic acid = 70:40:10:5),

Rf = 0.46 (n-butanol:acetic acid:water = 75:7.5:21).

The starting material is produced in the following way:

Step 25. 1

2.03 g (4 mmol) of N-acetyl-muramyl-L-α-aminobutyryl-D-isoglutamine (α,3 mixture) is transferred to the benzhydryl ester analogously to step 8.1. After normal work-up, N-acetyl-muramyl-L-cx-aminobutyryl-D-isoglutamine-benzhydryl ester (a,3-mixture) is obtained as a colorless powder;

[a]D° = +41.8 ± 1.8° (c = 0.548; methanol),

Rf = 0.73 (chloroform:methanol:water = 70:30:5),

Rf = 0.78 (acetonitrile water = 3:1).

Step 25. 2

1.70 g (2.53 mmol) N-acetyl-muramyl-L-α-aminobutyryl-D-isoglutamine-benzhydrylase (oc,3 mixture), dissolved in 25 ml abs. pyridine, is perpropionylated analogously to step 8.2 (90 equivalents of anhydride). After standing for 3 hours at RT, 30 ml of water is added and the whole is evaporated in a large HV at 30°C to dryness. The resinous residue is taken up in 80 ml abs.

dioxane, filtered (PTFE; 0.2 µm) and lyophilized. Man N -acetyl-1,4,6-tri-O-propionyl -mu ra myl -L- a - aminobutyryl-D -isoglutamine -benzhyd ryl ester (cx,p mixture) as colorless powder containing 0.79 mol of water;

[a]D° = +52.2 ± 3.7° (c = 0.270; methanol),

Rf = 0.67 (acetonitrile water = 3:1),

Rf = 0.32 (chloroform:isopropanol:acetic acid = 70:8:2),

Rf = 0.87 (ethyl acetate:n-butanol:pyridine:acetic acid:water =

42:21:21:6:10).

EXAMPLE 26

0.50 g (1 mmol) of N-acetyl-desmethylmuramyl-L-alanyl-D-glutamic acid diamide (cx, p-mixture) is peracetylated analogously to step 8.2. After normal work-up, the crude product is purified by chromatography on silica gel (1:80) in the system chloroform-methanol-water (70:30:5). The uniform fractions are collected, taken up in 50 ml of double distilled water, the solution is filtered ("Naglene", 0.2 µm)

and lyophilized. N-acetyl-1,4,6-tri-O-acetyl-desmethylmuramyl-L-alanyl-D-glutamic acid diamide (a,p mixture) is obtained as a colorless powder containing 1.69 mol of water;

[cOj^ = +36.2 ± 1.8 (c = 0.5; dimethylformamide),

Rf = 0.49 (chloroform:methanol:water = 70:30:5),

Rf = 0.20 (n-butanol:acetic acid:water = 75:7.5:21).

EXAMPLE 27

0.50 g (0.89 mmol) of N-benzoyl-desmethylmuramyl-L-alanyl-D-glutamic acid dimethyl ester (a,<p->mixture) is peracetylated analogously to step 8.2. The crude product is purified on silica gel (1:300; 3 ml fractions). The eluent used is n-butanol-acetic acid-water (75:7.5:21). The pure fractions are collected, dissolved in 10 ml of chloroform-methanol-water (70:30:5), filtered (PTFE; 0.2 µm) and lyophilized after evaporation and addition of 60 ml of abs. dioxane. 1,4,6-tri-O-acetyl-N-benzoyl-desmethylmur amyl-L-alanyl-D-glutamic acid dimethyl ester (cx,p mixture) is obtained in the form of a white powder containing 1.31 moles of water;

[cx]D<0>= +39.2 ±1°(c = 0.615; methanol),

Rf = 0.68 (n-butanol:pyridine:acetic acid:water = 62:21:6:11),

Rf = 0.53 and 0.57 (n-butanol:acetic acid:water = 75:7.5:21).

EXAMPLE 28

0.475 g (0.91 mmol) N-acetyl-desmethylmuramyl-L-alanyl-D-glutamic acid dimethyl ester (cx, p mixture) is peracetylated analogously to step 8.2 and purified by chromatography on silica gel (1:100; 2.5 ml fractions ) in the system chloroform-isopropanol (70:8). The pure fractions are collected, taken up in 60 ml abs. dioxane, filtered (PTFE; 0.2 µm) and lyophilized. N-acetyl-1,4,6-tri-O-acetyl-desmethylmuramyl-L-alanyl-D-glutamic acid dimethyl ester (cx,<p->mixture) is obtained as a colorless powder containing 0.95 mol of water.

[cx]j}° = +20.8 ±2°(c = 0.491; chloroform),

Rf = 0.55 (chloroform:methanol = 85:15),

Rf = 0.37 (n-butanol:acetic acid:water = 75:7.5:21).

EXAMPLE 29

To a solution of 0.200 g (0.28 mmol) of N-acetyl-muramyl-L-α-aminobutyryl-D-isoglutaminyl-L-arginine methyl ester hydrochloride (cx,p mixture) in 2 ml of abs. pyridine, 0.263 ml (2.8 mmol) of acetic anhydride is added and the mixture is allowed to stand overnight at RT. The reaction solution is mixed with 15 ml of water and evaporated in vacuo at 30 "C. The crude product is purified by chromatography on silica gel (1:100) in the system chloroform-methanol-water (70:30:5). The uniform fractions are collected, taken up in 3 ml double-distilled water, filtered (0.45 µm) and lyophilized after adding 35 ml of absolute dioxane.N-acetyl-1,4,6-tri-0-acetyl-muramyl-L-cx-aminobutyryl-D-isoglutaminyl is obtained - L-arginine methyl ester hydrochloride hydrate (a, p mixture) in the form of a colorless powder containing 2 moles of water;

Rf = 0.29 (chloroform:methanol:water = 70:30:5)

Rf = 0.60 (ethyl acetate:acetic acid:water:methanol = 67:10:23:12).

The starting material is produced in the following way:

Step 29. 1

To a solution of 2.35 g (4.64 mmol) N-acetyl-muramyl-L-cx-aminobutyryl-D-isoglutamine (a,p mixture) and 1.21 g (4.64 mmol) L-arginine -methyl ester dihydrochloride in 100 ml abs. dimethylformamide, 0.84 g (5.6 mmol) of 1-hydroxy-benztriazole monohydrate, 0.51 ml of N-methylmorpholine and finally 1.15 g (5.6 mmol) of dicyclohexylcarbodiimide are successively added. After 48 hours of stirring at RT, the yellow suspension is evaporated on a rotary evaporator in a high vacuum at 30"C and the residue is stirred after adding 200 ml of water. The precipitate is filtered off and the aqueous phase is extracted four times, each time with 50 ml of n-butanol which is saturated with water. The aqueous phase is then freed from butanol in vacuo and lyophilized. The residue is purified by chromatography on silica gel (1:100; 10 ml fractions) in the system chloroform-methanol-water (70:30:5). Still present arginine-methylester- hydrochloride is removed by reverse-phase chromatography ("Opti UPC12"; 40-63 µm; 1:42) with 0.5 liters of water, the product is then eluted with the same volume of acetonitrile-water (9:l). It is evaporated strongly in vacuo, the solution is sterile filtered (PTFE; 0.2 µm) and lyophilized to give N-acetyl-muramyl-L-cx-aminobutyryl-D-isoglutaminyl-L-arginine methyl ester hydrochloride (a,p mixture) as colorless powder containing 2.35 moles of water;

[a]j)<0>= +18.5 ± 0.8°(c = 1.224; methanol),

Rf = 0.29 (ethyl acetate: n-butanol: pyridine: acetic acid: water

= 42:21:21:6:10);

Rf = 0.22 (ethyl acetate:acetic acid:water:methanol = 67:10:23:12).

EXAMPLE 30

To 50 mg (0.074 mmol) N-acetyl-1,4,6-tri-0-propionyl-muramyl-L-cx-aminobutyryl-D-isoglutamine (a,3 mixture; see Example 15) dissolved in 0.5 ml abs. dimethylformamide, 13.2 mg (0.087 mmol) of 1-hydroxybenztrial (containing 11-13% water), 17.6 mg (0.067 mmol) L-arginine methyl ester dihydrochloride, 8 pl (0.071 mmol) N-methyl-morpholine and finally 20.8 mg (0.10 mmol) of dicyclohexylcarbodiimide. After 2 days of stirring at RT, the yellow suspension is evaporated to dryness and the residue is purified on silica gel (1:100) in the system ethyl acetate-n-butanol-pyridine-acetic acid-water (42:21:21:6:10) twice. One obtains N-acetyl-1,4,6-tri-O-propiony 1-muramyl-L-cx-aminobutyryl-D-isoglutami n-L-arginine methyl ester hydrochloride (cx, p-mixture ) as colorless powder;

Rf = 0.52 (ethyl acetate:n-butanol:pyridine:acetic acid:water = 42:21:21:6:10),

Rf = 0.40 (chloroform:methanol:water = 70:30:5).

EXAMPLE 31

0.080 g (0.085 mmol) Na<->(N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-alanyl-D-isoglutaminyl)-N^-benzyloxycarbonyl-L-4-thialysine isopropyl ester (cx, 3, mixture) is dissolved in 8 ml of methanol containing 25 ml of 30% hydrogen peroxide solution, and allowed to stand for 2 days at RT. The excess of hydrogen peroxide is decomposed by adding platinum on carbon, the latter is filtered off and the filtrate is evaporated to dryness. The residue is taken up in 90% dioxane, filtered (PTFE; 0.2 µm) and lyophilized. One obtains N^-(N-acetyl - 1,4,6-tri-O-acetyl-muramyl-D-is oglutami nyl)-N^- benzyloxycarbonyl-L-4-thia-lysine-isopropylester- sulfoxide (cx, 3 mixture) as colorless powder.

Rf = 0.64 (chloroform:methanol:water:acetic acid = 70:40:10:5),

Rf = 0.75 (ethyl acetate: n-butanol: pyridine: acetic acid: water

= 42:21:21:6:10).

The starting material is produced in the following way:

Step 31. 1

To a solution of 0.31 g (0.81 mmol) of benzyloxycarbonyl-L-thialysine isopropyl ester hydrochloride and 0.089 ml (0.81 mmol) of N-methyl-morpholine in 3.5 ml of abs. 0.82 g (0.97 mmol) of N-acetyl-muramyl-L-alanyl-D-isoglutamine-N-hydrosuccinimide ester [cx, 3-mixture] is added to dimethylformamide; about. 70%, otherwise dicyclohexylurea] and the whole is stirred for 16 hours at RT. The suspension is mixed with 20 ml of acetic acid ethyl ester, stirred for one hour at 0°C and the insoluble dicyclohexylurea is suctioned off. The residue remaining after evaporation is taken up in 50 ml of n-butanol saturated with water and extracted six times, each time with 10 ml of n-butanol saturated with water. The upper phases are collected, evaporated to approx. 5 ml and the whole is lyophilized after adding 45 ml abs. dioxane. Na-(N-a cety 1-mu ramy 1-L-alanyl-D-isoglutaminyl)-N £-benzyl oxycarbonyl-L-thialysine-isopropyl ester (a,3-mixture) is obtained as a colorless powder which still contains some dicyclohexylurea;

Rf = 0.46 (ethyl acetate:acetic acid:water:methanol = 67:10:23:12),

Rf = 0.69 (acetic acid ethylase:n-butanol:pyridine:acetic acid:water

= 42:21:21:6:10).

Step 31. 2

0.38 g (0.47 mmol) Na<->(N-acetyl-muramyl-L-alanyl-D-isoglutaminyl)-N<£->benzyloxycarbonyl-L-thialysine isopropyl ester (a,p mixture), dissolved in a mixture of 0.40 ml abs. dimethylformamide and 4 ml abs. pyridine is mixed with 0.20 ml (2.1 mmol) of acetic anhydride and allowed to stand for 1 1/2 days at RT. The reaction solution is evaporated and purified by chromatography on silica gel (1:200) in the system chloroform-methanol (9:1). The unified fractions are gathered. One obtains Na<->(N-acetyl-1,4,6-tri-0-acetyl-muramyl-L-alanyl-D-isoglutaminyl)-N E -b enzylo xyc arbo nyl- L-thi alys in-i sopr opyl ester r-hy drate (cx, p mixture) as colorless powder;

Rf = 0.90 (chloroform:methanol:water:acetic acid = 55:47:13:5),

Rf = 0.81 (ethyl acetate: n-butanol: pyridine: acetic acid: water

= 42:21:21:6:10),

Rf = 0.39 (n-butanol:acetic acid:water = 75:7.5:21).

EXAMPLE 32

Analogous to example 2, one obtains from 1.35 g (1.61 mmol) N-propionyl-1,4,6-tri-O-propionyl-desmethylmuramyl-L-al anyl-D-isoglutamine-(C^) -benzhydryl ester containing 0.68 mol of water, by catalytic hydrogenation with 0.2 g of 10% palladium carbon in 50 ml of methanol-tetrahydrofuran (1:1) after lyophilization N-propionyl-1,4,6-tri-O-propiony 1 -desmethylmuramy 1-L-alanyl-D-isoglutamine (cx,p mixture) as colorless powder containing 1.55 moles of water;

[cx]D° = +41.7 ± 2.1" (c = 0.484; water),

Rf = 0.20 (chloroform:methanol = 7:3),

Rf = 0.31 (chloroform:methanol:water = 70:30:5).

EXAMPLE 33

Analogous to example 28, 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid diethyl ester (cx,<p->mixture) is obtained.

EXAMPLE 34

1.45 g (1.76 mmol) N-acetyl-1,4,6-tri-O-propionyl-muramyl-L-alanyl-D-isoglutaminyl-L-alanine benzyl ester (a,p mixture), dissolved in 100 ml of glacial acetic acid, hydrated as usual in the presence of palladium on carbon (20%), filtered and

the solution is lyophilized. The residue is taken up in 50 ml of double-distilled water and filtered ("Nalgene"; 0.45 pm). After lyophilization, N-acetyl-1,4,6-tri-O-propionyl-muramyl-L-alanyl-D-isoglutaminyl-L-alanine (cx, g mixture) is obtained as a colorless powder containing 1.52 mol of water ;

and[cx]D° = +42.2 ± 1.5°(c = 0.657; methanol),

Rf = 0.15 (chloroform:methanol:water = 70:30:5),

Rf = 0.26 (n-butanol:acetic acid:water = 75:7.5:21),

Rf = 0.42 (ethyl acetate: n-butanol: pyridine: acetic acid: water

= 42:21:21:6:10).

The starting material is obtained in the following way:

Step 34. 1

1.46 g (2.23 mmol) of N-acetyl-muramyl-L-alanyl-D-isoglutaminyl-L-alanine benzyl ester (cx, g mixture) is dissolved in 25 ml of abs. pyridine and after the addition of 1.74 g (13.4 mmol) is allowed to stand at RT for 36 hours. The solution is evaporated in high vacuum at 30 °C, the residue is taken up in 50 ml dioxane-water (1:2), filtered (PTFE; 0.2 µm) and lyophilized. N-acetyl-muramyl-L-alanyl-D-isoglutaminyl-L-alanine benzyl ester (a,<g->mixture) is obtained as a colorless powder containing 0.73 mol of water;

[cx]D° = +38 ± 1.1<*>(c = 0.900; glacial acetic acid),

Rf = 0.69 (chloroform:methanol:water = 70:30:5),

Rf = 0.33 (chloroform:methanol:water:acetic acid = 85:13:1.5:0.5).

EXAMPLE 35

1.00 g (0.99 mmol) Na<->(N-acetyl-tri-O-propionyl-muramyl-L-alanyl-D-isoglutaminyl)-N^-benzyloxycarbonyl-L-lysine benzyl ester (a, g <->mixture) is hydrated analogously to example 34 in glacial acetic acid. The lyophilisate obtained by normal processing is thoroughly triturated with 30 ml abs. diethyl ether. The precipitate is filtered off, taken up in 50 ml of double-distilled water, filtered ("Nalgene"; 0.45 µm) and lyophilized. N-acetyl-1,4,6-tri-0-propionyl-muramyl-L-alanyl-D-isoglutaminyl-L-lysine (cx, g<->mixture) is obtained as a colorless powder containing 3.1 mol of water ;

[cx]D° = +49.7 ± 1.1"(c = 0.903; methanol),

Rf = 0.33 (ethyl acetate:acetic acid:water:methanol = 67:10:23:12),

Rf = 0.21 (chloroform:methanol:water:acetic acid = 70:40:10:0.5),

Rf = 0.08 (ethyl acetate: n-butanol: pyridine: acetic acid: water

= 42:21:21:6:10).

The starting material is produced in the following way:

Step 35. 1

To a suspension consisting of 8.10 g (20 mmol) of benzyloxycarbonyl-L-lysine benzyl ester hydrochloride and 17.30 g (20 mmol) of N-acetyl-muramyl-L-alanyl-D-isoglutamine-N-hydroxy- succinimide ester (cx, p-mixture; approx. 70%, otherwise dicyclohexylurea) in 180 ml of dimethylformamide, 2.02 g (20 mmol) of triethylamine, dissolved in 20 ml of dimethylformamide, is added dropwise with good stirring over the course of 15 minutes. After 10 hours of stirring at RT, the insoluble "urea" is filtered off, the filtrate is evaporated to approx. 30 ml and the product is precipitated by portionwise addition of a total of 300 ml of water. The precipitate is suctioned off, taken up after drying in 50 ml of dimethylformamide and precipitated again in the same way with 500 ml of acetic acid ethyl ester (saturated with water). The crude product (13.4 g) is purified by chromatography on silica gel (1:100) in the system chloroform-methanol-water (70:30:5). The pure fractions give Na<->(N-acetyl-muramyl-L-alanyl-D-isoglutaminyl)-N^-benzyloxycarbonyl-L-lysine benzyl ester (cx,p mixture) containing 1.28 moles of water;

Skirn = +19 ±1° (c = 0.564; glacial acetic acid),

Rf = 0.47 (chloroform:methanol:water = 70:30:5),

Rf = 0.40 (n-butanol:acetic acid:water = 75:7.5:21),

Rf = 0.77 (ethyl acetate: n-butanol: pyridine: acetic acid: water

= 42:21:21:6:10).

Step 35. 2

2.00 g (2.37 mmol) Na<->(N-acetyl-muramyl-L-alanyl-D-isoglutaminyl)-NE<->benzyloxycarbonyl-L-lysine benzyl ester (cx,P mixture) is dissolved in 25 ml abs. pyridine and is perpropionylated analogously to step 34.1. After standing for 30 hours at RT, it is evaporated at 30 °C in HV and the residue is purified by chromatography on silica gel (1:100) in the system chloroform-methanol-water (70:30:5). The unified fractions are gathered. One obtains ^-(N-acetyl-tri-O-propionyl-muramyl-L- alanyl-D -isoglutaminyl)-N ^-benzyloxycarbonyl-L-

lysine benzyl ester (α,3 mixture) as colorless powder containing 0.56 moles of water;

[a]D° = +28.8 ± 1.0° (c = 0.983; methanol),

Rf = 0.90 (chloroform:methanol:water = 70:30:5),

Rf = 0.11 (chloroform:isopropanol:acetic acid = 70:8:2).

Claims (10)

1. Process for the preparation of compounds of formula I in which the hexose moiety is derived from D-glucose, D-mannose or D-galactose, n stands for 0 or 1 and R <*> stands for lower alkanoyl or benzoyl, R <2> stands for lower alkyl or phenyl, R <3> , R <5> and R <7> independently stand for hydrogen or lower alkyl, or R <5> and R <8> together stand for trimethylene and R <7> stand for hydrogen, R <4> and R <6 > independently stand for hydrogen, lower alkanoyl or benzoyl, R <8> stands for hydrogen or unsubstituted or lower alkyl substituted with phenyl, hydroxy, mercapto or lower alkylthio, R <9> and R <*2> stand independently for hydroxy, amino or lower alkoxy, R <lO> represents hydrogen, carboxy or lower alkoxycarbonyl and R^ represents hydrogen or unsubstituted or with amino, lower alkanoylamino, hydroxy, guanidino, lower alkanoyloxy, 2-benzyloxycarbonylaminoethyl-sulfinyl,, 2-benzyloxycarbonylaminoethyl-sulfonyl, 2- lower alkoxycarbonylaminoethylsulfinyl or 2-lower alkoxycarbonylaminoethylsulfonyl subs titrated lower alkyl, under the condition that in the compounds in which the pyranose part is derived from D-glucose and at the same time n stands for 0, R^, R <7> and R^ stand for hydrogen, R <8> for methyl, R <9> for amino and R <12> for hydroxy and in which the residues R <*> , R <4> and R <6> all three have the same meaning, R <*> , R <4> and R <6> are different from acetyl and butyryl, when R <2> stands for phenyl and at the same time R <3> stands for hydrogen, and that R <1> , R <4> and R <6> are different from acetyl when R <2> and R <3> both stand for methyl, and salts of such compounds with at least one salt-forming group, characterized in that mana) reacts with a compound of formula II in which at least one of the radicals R^ <a> , R <2a> and, for the preparation of compounds of formula I, in which R <4> and R <6> have the meanings given above with the exception of hydrogen, R <4a> and R <6a> stands for hydrogen and the other residues have the meanings of R^, the group R <2-> C(=0)-, R <4> respectively. R <6> , and the rest of the substituents have the above-mentioned meanings, whereby in a compound of formula II present free functional groups, with the exception of the groups which will participate in the reaction, are protected if necessary with easily cleavable protective groups, with an acylating agent which transfers the acyl residue R <1> , R <2-> C(=0)-, R <4> or R <6> to be introduced and, if necessary, cleaves off any protective groups present, or b) reacts with a compound of formula III in which q, r, s and t independently stand for 0 or 1 and in which the substituents have the meanings indicated above, whereby in a compound of formula V present free functional groups, with the exception of the groups which are to participate in the reaction, can, if necessary be protected with easily cleavable protecting groups, or a reactive carboxylic acid derivative thereof, with a compound of formula VI in which u, v and x independently of each other stand for 0 or 1 and the other symbols and substituents have the meanings indicated above, whereby in a compound of formula VI present free functional groups, with the exception of the groups which will participate in the reaction, if necessary may be protected by easily removable protecting groups, whereby u, v and x stand for 1 when q and t stand for 0 in the reaction partner of formula V, or u stands for 0 and v and x stand for 1 when q stands for 1 and t stands for 0, or u and v stand for 0 and x stands for 1 when q, r and t stand for 1 and s stands for 0, or (for the preparation of compounds of formula I in which n stands for 1) u and x stands for 0, when q, r, s and t stand for 1, or a reactive functional derivative thereof, and if necessary cleaves off protecting groups present, or d) for the preparation of a compound of formula I, in which R <9> has one of the above-mentioned meanings except hydroxy, and the other substituents have the above-mentioned meanings, amidates or esterifies a compound of formula VII in which R<13> stands for carboxy, and in which the other substituents have the above-mentioned meanings, whereby in a compound of formula VII present free functional groups, with the exception of R<13>, are, if necessary, protected with easily removable protective groups , or a reactive carboxylic acid derivative thereof, <1> and if necessary cleaves off any protective groups present, or e) in a compound of formula I, in which at least one of the residues OR <4> , OR <6> , R <8> , C(=0)-R 9,R 10,R 1 <1> and C( = 0)-R12 is present in a protected form, which does not correspond to the definition for the desired end product, splits off the relevant protective groups, and, if desired, after carrying out one of the methods a-e) transfers a prepared compound of formula I with at least one salt-forming group to its salt or converts a prepared salt of a compound of formula I to the free compound, and/or splits a prepared isomer mixture. 2. Method according to claim 1, characterized in that starting materials are selected in such a way that a compound of formula I is produced in which the hexose part is derived from D-glucose or D-mannose, n stands for 0, R <*> , R <4 > and R <6> stand independently of each other for C2 _5~ alkanoyl or benzoyl, R <2> stands for <C>j _4 -alkyl or phenyl, R <3> , R^ and R <7> stand independently of each other for hydrogen or methyl, or R^ and R<8> together stand for trimethylene and R<7> stands for hydrogen, R<8> for hydrogen, Cj_4 -alkyl or C1_2 -alkyl substituted with phenyl, hydroxy, mercapto or methylthio . N-benzoyl-1,4,5-tri-0-butyryl-desmethylmuramyl-L-alanyl-D-isoglutamin, N-benzoy 1-1,4,5-tri-O-acethyl-desmethyl-muramyl -L-alanyl-D-isoglutamine and N-acetyl-1 ,4,6-tri-O-acetyl-muramyl-L-alanyl-D-isoglutamine and salts thereof. 3. Process according to claim 1, characterized in that starting materials are chosen in such a way that a compound of formula I is produced, in which the hexose part is derived from D-glucose, n stands for 1, Ri, R<4> and R° stand independently independently of each other for C2-5~alkanoyl or benzoyl, R<2> stands for Cj_4 -alkyl or phenyl, R3,R^ and R<7> independently of each other stand for hydrogen or methyl, or R^ and R<8> together stand for trimethylene and R <7> stand for hydrogen, R <8> for hydrogen, Cj_4 -alkyl or with phenyl, hydroxy, mercapto or methylthio substituted C^ _2~ alkyl, R <9> and R <12> independently stand for hydroxy, amino or lower alkoxy, R <10> stands for hydrogen, carboxy or lower alkoxycarbonyl and RH stands for unsubstituted or with amino, guanidino, lower alkanoyloxy, 2-benzyloxycarbonylaminoethylsulfinyl, 2-benzyloxycarbonylaminoethylsulfonyl, 2-lower oxycarbonylaminoethylsulfinyl or 2- lower alkoxycarbonylaminoethylsulfonyl substituted lower alkyl, or a salt t of such a compound with at least one salt-forming group. 4. Method according to claim 1, characterized in that one chooses the starting material in such a way that a compound of formula I is prepared, in which the hexose part is derived from D-glucose, n stands for 0 or 1, and Ri stands for C2 -5~ alkanoyl or benzoyl, R <2> stands for Cj_4 -alkyl or phenyl, R <3> stands for hydrogen or C^ _4~ alkyl, R <4> stands for hydrogen, C2 _5~ alkanoyl or benzoyl, R^ stands for hydrogen, methyl or together with R <8> for trimethylene, R° stands for C2 _5 -alkanoyl or benzoyl, R <7> stands for hydrogen or methyl, R <8> stands for Ci_4~ alkyl or together with R <5> for trimethylene, R <9> stands for hydroxy, lower alkoxy or amino, R <10> stands for hydrogen or carboxy, RH stands for unsubstituted or with amino, guanidino or 2-benzyloxycarbonylaminoethylsulfinyl substituted Cj_4 -alkyl and R <l2> stands for hydroxy, lower alkoxy or amino, or a salt of such a compound with at least one salt-forming group with the exception of N-benzoyl-1,4,6-tri-O-butyryl-desmethylm uramyl-L-alanyl-D-isoglutamine, N-benzoyl-1,4,6-tri-0-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine and N - acetyl -1,4,6-tri-O - acetyl - mu r amyl - L - alanyl - D - isoglutamine and salts thereof. 5. Process according to claim 1, characterized in that one chooses the starting material in such a way that a compound of formula I is produced, in which the hexose part is derived from D-glucose, n stands for 0 or 1, Ri stands for C2_5 -alkanoyl or benzoyl , R <2> stands for Cj_2~ alkyl or phenyl, R <3> stands for hydrogen or methyl, R <4> stands for hydrogen, C2 _5 -alkanoyl or benzoyl, R^ stands for hydrogen, R° stands for C2 _5 ~ alkanoyl or benzoyl, R <7> stands for hydrogen, R <8> stands for methyl, ethyl or 2-propyl, R <9> stands for hydroxy, amino or Cj_4 -alkoxy, R <lO> stands for hydrogen, RH stands for methyl or 4-amino-n-butyl and R <l2> stands for hydroxy, amino of Cj_4~ alkoxy, or a salt of such a compound with at least one salt-forming group with the exception of N-benzoyl-l,4,6 -tri-0-butyryl-desmethylmuramyl-L-alanyl-D-isoglutamine, N-benzoyl-1,4,6-tri-0-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine and N-acetyl-1,4 ,6-tri-O-acetyl-muramyl-L-alanyl-D-isoglutamine and salts thereof. 6. Method according to one of the claims 1-5, characterized in that starting materials are selected in such a way that a compound of formula I is produced, in which at least one of the residues R <9> and R <*2> stands for lower alkoxy and the other of the residues R <9> and R^ <2> stand for hydroxy, amino or lower alkoxy, or a salt of such a compound with at least one salt-forming group. 7. Process according to one of claims 1-6, characterized in that the starting material is selected in such a way that a compound of formula I is produced, in which n stands for 1 and R^ is different from hydrogen, or a salt of such a compound with at least one salt-forming group. 8. Process according to one of claims 1-7, characterized in that the starting materials are selected in such a way that a compound of formula I is produced, in which R <8> stands for C2 _4~ alkyl or, together with R <5>, stands fort rimethylene, or a salt of such a compound with at least one salt-forming group. 9. Process according to claim 1, characterized in that the starting materials are selected in such a way as to produce (1a,a),4,6-tri-O-ac ethyl-N-propionyl-desmethylmuramyl-L-alany l-D-glutamic acid-(Ca ) - n-butyl ester or a pharmaceutically usable salt thereof. 10. Process according to claim 1, characterized in that the mother chooses the starting materials in such a way that N-acetyl-1,4,6-tri-O-propionyl-mur amyl-L-alany 1-D-isoglutaminyl-L-alanine is produced or a pharmaceutically acceptable salt thereof.