NO860624L - ACYLATED HEXOS DERIVATIVES AND PROCEDURES FOR PREPARING THEREOF. - Google Patents

Description

The present invention relates to 1,4,6-tri-O-acylated mura-mylpeptide 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

wherein the hexose moiety is derived from D-glucose, D-mannose or D-galactose, n stands for 0 or 1, and R 1 , R 4 and R < 6> independently stand 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 mean trimethylene and R 7 stand for hydrogen,

R Q for hydrogen or unsubstituted or with phenyl, hydroxy, mercapto or lower alkylthio-substituted lower alkyl, R<9>

and R 12 independently of each other stand for hydroxy, amino, C^_-^Q-alkoxy, aryllaverealloxy, alkanoyloxyllaverealloxy with up to 16 C atoms, aroyloxyllaverealloxy, 3-cholesteryloxy or 2-trimethylamino-ethyloxy, R stands for hydrogen, carboxy, lower alkoxycarbonyl1 or aryl or lower alkoxycarbonyl and R stands for hydrogen or unsubstituted or with amino, hydroxy, lower alkanoylamino, lower alka-

noyloxy, 2-benzyloxycarbonylamino-ethyl-sulfonyl, 2-benzyloxycarbonylamino-ethyl-sulfonyl, 2-lower oxycarbonyl-1-amino-ethyl-sulfinyl, 2-lower oxycarbonylamino-ethyl-sulfonyl or guanidino substituted lower alkyl, provided that at least one of the residues R and R<12> are different from hydroxy, amino and C-12-Alkoxy or R from hydrogen, carboxy and alkoxycarbonyl with up to 7 C atoms in the alkoxy part, and salts of such compounds with at least one salt-forming group .

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

In cases.asymmetric substitution is the configuration

3 8 9

at the atoms C-R , C-R respectively C-CO-R (D), (L) respectively (D), as indicated in formula I. The configuration at C-R^ is in the case of asymmetric substitution (L) or

(D), preferably (L).

14 6

Lower alkanoyl R , R and R are especially C 2-6 alkanoyl, e.g. n-hexanoyl, preferably C 2-4 -alkanoyl, e.g. butyryl, propionyl or preferably acetyl.

Lower alkyl R 2 is preferably C 14 , especially C 12 alkyl.

3 5 7

Lower alkyl R , R or R is preferably C-^_^-alkyl, especially methyl.

8 11

Unsubstituted lower alkyl R or R preferably

alkyl, e.g. ethyl, isopropyl, 2-methylpropyl, sec.butyl or especially methyl.

With phenyl, hydroxy, mercapto or lower alkylthio, such as methylthio in particular, lower alkyl R substituted preferably gives correspondingly substituted C 2 -alkyl, e.g. benzyl, hydroxymethyl, 1-hydroxyethyl, mercaptomethyl or 2-methylthioethyl.

9 12

Alkoxy R or R is preferably C 1-3 alkoxy, e.g. methoxy, n-butyloxy or tert-butyloxy.

Aryl as part of aryl lavereal oxy, aryl lavereal oxycarbonyl or aroyloxylavereal oxy is particularly unsubstituted or with one or more, e.g. 1-3 of the aryl substituents mentioned below substituted phenyl or naphthyl, preferably unsubstituted phenyl.

Aryl substituents are especially lower alkyl, e.g. methyl, phenyl, halogen, hydroxy, lower alkoxy, e.g. methoxy, lower alkanoyloxy, e.g. acetoxy, amino, mono- or di-lower alkylamino, e.g. mono- or dimethylamino or lower alkanoylamino, e.g. acetylamino.

9 12

Aryl lavereal oxy R and/or R is with one or more, e.g. 1-3, "preferably 1 or 2, aryl radicals substituted by lower alkoxy, e.g. arylmethoxy, such as especially benzyloxy or benzhydryloxy.

Aryl lavereal oxycarbonyl R is with one or more, e.g. 1-3 preferably 1 or 2, aryl residues substituted lower alkoxycarbonyl, e.g. arylmethoxycarbonyl, such as especially benzyloxycarbonyl or benzhydryloxycarbonyl.

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

With amino or lower alkanoylamino substituted lower alkyl' R 1 is preferably 4-amino-n-butyl 1 or 4-lower alkanoylamino-n-butyl. With 2-benzyloxycarbonylamino-ethyl-sulfinyl

-or -sulfonyl or with 2-lower oxycarbonylamino-ethyl-sulfinyl or -sulfonyl substituted lower alkyl R"^, is preferably correspondingly substituted methyl, e.g. CgH5-CH2-0-C(=0)-NH-CH2-CH2- S(=0)-CH2- or CgHg-CI^-O-C(=0)-NH-CH2-CH2-SO2-CH2-. With guanidino substituted

lower alkyl R is preferably 3-guanidiono-n-propyl.

Alkanoyloxy methoxy R9 : or R 12 is particularly lower alkanoyloxy methoxy, e.g. pivaloyloxymethoxy.

9 12

Aroyloxyvareal oxy R or R is especially aroyloxy-methoxy, e.g. benzoyloxymethoxy.

With hydroxy or lower alkanoyloxy substituted lower alkyl R~ is especially correspondingly substituted C 2 -alkyl, e.g. hydroxymethyl, 1-hydroxy-ethyl, lower alkanoyloxymethyl rents 1-loweralkanoyloxyethyl.

3-cholesteryloxy is the radical formed by the removal of hydrogen from the hydroxy group of cholesterol.

The general designations used in the above and following preferably have the following meanings: Residues denoted by the prefix "lower" preferably 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 or guanidino groups. Compounds of formula I which exhibit a trimethylammonio group exist in salt form. Depending on the type of the salt-forming group, the compounds of formula I form with sodium 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 sulphites, 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 come into question for the formation of salts, such as lower alkylamines, 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-diethylaminoethyl ester, lower alkylene amines, e.g. 1-ethylpiperidine, cycloalkylamines, e.g. dicyclohexylamine, or benzylamine, e.g. N,N'-dibenzylethylenediamine, further bases of the pyridine type, e.g. idine, 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 organic carboxylic or sulphonic acids, e.g. trifluoroacetic acid, as well as with amino acids,

such as arginine or lysine. In the presence of several acidic or basic groups, mono- or polysalts can be formed. Compounds of formula I with an acid, e.g. free carboxyl group, a free basic, e.g. an amino group, can also exist in the form of smaller salts, i.e. in doubly ionic form. Or part of the molecule can exist as an inner salt and another part as a normal salt.

Pharmaceutically unsuitable salts can also be used for isolation or purification. However, only the pharmaceutically usable, non-toxic salts, which are therefore preferred, come to therapeutic use.

From the French patent application 74 22 909 with the publication number . 2,292,486, muramyl peptides of the type for the compound N-acetyl-muramyl-L-alanyl-D-isoglutamine ("MDP") are known. These compounds are described as immunological adjuvants, i.e. they can be used in a mixture with vaccination substances to improve the outcome of the vaccination. Own investigations, e.g. with mice infected with influenza virus,

have shown that N-acetyl-muramyl-L-alanyl-D-isoglutamine per

see, i.e. without the addition of vaccination substances is ineffective in the prophylaxis and therapy of viral infections. Furthermore, the applicability of the structurally simpler muramyl peptides

ene and their analogous compounds against viral infections not yet described.

The present invention is based on that task

to provide structurally relatively simple and consequently easily produced muuramyl peptide derivatives, which after administration to warm-blooded mammals are very effective when it comes to prophylaxis and therapy for 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 for therapy in viral infections, which e.g. appears from animal experiments, as stated in the example section. In these animal experiments, animals, such as mice or guinea pigs, are infected with different virus species in a dose which for all or the great majority of untreated (control) animals is fatal, e.g. LDgQ_<g>Q, and the course of the infection is observed in untreated control animals in comparison with animals which before, together with or after the infection have been treated with one of the above-mentioned compounds or a salt thereof.

Thereby it turns out that a prophylactic effect occurs

when administering the compounds of formula I already several days up to several, e.g. four weeks before the infection and a therapeutic effect is achieved by administration over several days, e.g. 1 week after the infection.

The compounds of formula I are in the above-mentioned test

on mice already effective in the dose range between 0.001 mg/kg and 0.1 mg/kg.

Also noteworthy is the broad viral spectrum, on the other hand

the above-mentioned compounds are effective.

The compounds of formula I can in particular be used in the prophylaxis and therapy of diseases caused by the viruses mentioned below (for nomenclature see J.L. Melnick, Prog. med. Virol, 2j>, 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 also 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 enveloped and bare virion, and in the case of RNA viruses with helical symmetry for the capsid, in which the nucleocapsidome envelope is located at the surface membrane, but also in the case of adenoviruses, poxviruses and coronaviruses, especially in the case of human coronaviruses.

First of all, the compounds of formula are used

In the case of herpesviruses, picornaviruses and myxoviruses, but also in the case of mastadenoviruses, especially human-like adenoviruses, in the case of chordopoxviruses, mainly orthopoxviruses, which especially e.g. vacciniavirus, in the case of reoviruses, primarily (especially human) rotaviruses, similar to caliciviruses and rhabdoviruses, primarily human vesiculoviruses, as well as horses, bulls and pigs.

The compounds of formula I are mainly used in conjunction with alpha herpesviruses, such as varicella viruses, e.g. human varicella-zoster viruses, rhinoviruses, cardioviruses and orthomyxoviruses, but also in cases of betaherpesviruses, such as especially human cytomegaloviruses, in the case of aphtoviruses, primarily ungulate aptoviruses, mainly bulls, as well as in cases of paramycoviruses, primarily pneumoviruses, f .ex. respiratory syncytial viruses in humans, and also morbilliviruses and paramyxoviruses, such as parainfluenza viruses, e.g. human parainfluenza virus, including sendaivirus as well as in the case of arboviruses or vesiculoviruses, e.g. vesicular stornatitis virus.

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

The compounds of formula I can be used for prophylaxis and therapy in viral infections, especially in warm-blooded mammals including humans, by being administered enterally or parenterally, primarily together with suitable auxiliary and carrier substances. Preferably they are applied to the mucous membrane, e.g. intranasally, rectally, vaginally or on the conjunctiva of the eye. However, the antivirus effect also occurs with other types of administration, e.g. subcutaneously, intravenously, intramuscularly, or by application to the normal skin.

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

For prevention, a single dose of approx. 0.01

mg to approx. 10 mg, preferably 0.05 to 1 mg, e.g. 0.2

mg, 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 extends over several weeks. If necessary, e.g. in times of high risk of infection, the administration of this dose can be repeated.

The therapeutic dose for warm-blooded animals with approx. 70 kg body weight is between 0.1 mg and 25 mg, preferably between 0.1 and 1 mg, e.g. at 0.5 mg, especially when administered orally. 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 occurs. Often, however, a single supply is sufficient.

The compounds of formula I also have anti-tumor properties. These depend on the ability of the connections, e.g. incorporated in aminar liposomes or in phosphate-buffered physiological saline solution (PBS), to activate macrophages in such a way that the body's own defense cells are able to kill tumor cells (cytotoxicity) or to prevent their growth (cytostasis). The induction of tumoricides and tumoristatic alveolar macrophages in rats in vitro and in situ can be 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 (intravenously or intranasally, in situ activation), or by a 24-hour pre-incubation with a compound of formula I in CO 2 incubators (in vitro activation). The activated macrophages are incubated for a further 72 hours with tumor cells.

To measure the tumoricidal activity of the macrophages, mer-125 the parent cells before the 72 hour incubation with I-iodo-deoxyuridine. The tumor cells that have not been killed can be measured after washing away the radioactivity released by the lysed tumor cells on the basis of the remaining radioactivity.

To determine the tumoristatic activity of the macrophages, 3H-thymidine is added to the cultures 8 hours before the deposition of the 72-hour incubation and then the 3H-thymidine incorporation into the tumor cells is measured. In vitro, the substances, both dissolved in TBS and also incorporated in liposomes already in doses of 20 ng/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 tumoricidal and tumoristatic alveolar macrophages. Furthermore, a single intranasal administration of the substance in PBS at a dose of 25 µg/rat causes the induction of tumoricidal alveolar macrophages.

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

The invention relates in particular to such compounds of formula I in which the hexose part is derived from D-glucose, D-mannose

o 14 or D-galactose, where n stands for 0 or 1, and R , R

and R 6 independently represent lower alkanoyl or benzoyl, R represents lower alkyl or phenyl, R 3 , R 5 and R 7 independently represent hydrogen or lower-5 8 7

alkyl, or R and R together for trimethylene and R for hydrogen, R g for hydrogen or unsubstituted or with phenyl, hydroxy, mercapto or lower alkylthio substituted lower alkyl, R<9> and R 12 stand independently of each other for hydroxy, amino, C^ _^q- alkoxy, aryl lavereal oxy, alkanoyloxylavereal oxy with up to 16 C atoms, aroyloxylavereal oxy or 3-cholesteryloxy, R stands for hydrogen, carboxy, lower alkoxycarbonyl or aryl lavereal oxycarbonyl,

and R^ stands for hydrogen or unsubstituted or amino- or hydroxy-substituted lower alkyl, under the ratio

rule that at least one of the residues R and R is different from hydroxy, amino and C^_^-alkoxy, or R"^ is different from hydrogen, carboxy and alkoxycarbonyl with up to 7 C atoms in the alkoxy part, and salts of such compounds with at least one salt-forming group.

Preferred are the above-mentioned compounds of formula

In which R 9 and R 10 independently of each other stand for hydroxy, amino, C^_^Q-alkoxy, alkanoyloxylaverealalkoxy with up to 16 C-atoms, 3-cholesteryloxy or unsubstituted in the phenyl part. pea or with lower alkyl, e.g. methyl, phenyl, halogen, hydroxy, lower alkoxy, e.g. methoxy, lower alkanoyloxy, e.g. acetoxy, amino, mono- or dilower alkylamino, e.g. mono- or dimethylamino, or lower alkanoylamino, e.g. acetylamino, substituted phenyl- or benzoyloxy-lower alkoxy, and R1(^ stands for hydrogen, carboxy, lower alkoxycarbonyl or in the phenyl part unsubstituted or with lower alkyl, e.g. methyl, phenyl, halogen, hydroxy, lower alkoxy, e.g. methoxy, lower alkanoyloxy, e.g. acetoxy, amino, mono- or dilower alkylamino, e.g. mono- or dimethylamino, or lower alkanoylamino, e.g. acetylamino, substituted phenyl lower real oxycarbonyl, and the other substituents have the meanings mentioned above, provided that that at least one of the residues R 9 and R 12 is different from hydroxy, amino and lower alkoxy or R 1 from hydrogen, carboxy and lower alkyl carbony 1, and salts of such phor compounds with at least one salt-forming group.

Particularly preferred are compounds of formula I in which the hexose moiety: is derived from D-glucose or D-mannose, n stands for 0 or 1, r\ R^ and R^ stand independently of each other for C__.-alkanoyl or benzoyl, R stands for for C 1 -alkyl

357o<14>

or phenyl, R , R and R independently stand for

5 8

hydrogen or methyl, or R and R together form trimethylene and R 7 stands for hydrogen, R<8> for hydrogen, C^_^-alkyl or C^_2-alkyl substituted with phenyl, hydroxy, mercapto or methylthio, R 9 and R 12 independently of each other stands for hydroxy, amino, lower alkoxy, phenyl lower oxy, lower alkanoyloxy lower oxy, benzoyloxy lower oxy or tri-cholesteryloxy, R"^ stands for hydrogen, carboxy, lower alkoxycarbonyl or phenyl lower oxycarbonyl and R^ stands for hydrogen or unsubstituted or with amino or hydroxy substituted C, .-alkyl, below the predicted-1 12

nmg that at least one of the residues R and R is different from hydroxy, amino or lower alkoxy, or R"^ from hydrogen,

carboxy and lower alkoxycarbonyl, 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 or D-mannose, n stands for 0 or 1, R 1, R 4 and R 6 stand independently of each other for C»-.-alkanoyl or benzoyl, R 2 stands for C, -alkyl

357 o

or phenyl, R , R and R independently stand for

9 12 hydrogen or methyl, R for C-^^-alkyl, R and R independently of each other for hydroxy, amino, C^_^- alkoxy, phenylmethoxy, lower alkanoyloxymethoxy, benzoyloxymethoxy or 3-cholesteryloxy, R"^ stands for hydrogen, carboxy, alkoxycarbonyl with up to 5 C atoms or phenylmethoxycarbonyl and R stands for C, .-alkyl, on the condition that at least 9 12 one of the residues R and R is different from hydroxy, amino and C^_^-alkoxy, or R^~ ® from hydrogen, carboxy and alkoxycarbonyl with up to 5 C atoms, 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, r\ R^ and R^ stand for acetyl or butyryl, R2 for C,__-3 5 7 alkyl or phenyl, R for hydrogen or methyl, R and R 8 9 for hydrogen, R for C1-3-alkyl, R for amino, alkoxy, pivaloyloxymethoxy or mono- or diphenylmethoxy, R10 for hydrogen, R for methyl and R12 for mono- or diphenylmethoxy or 3-cholesteryloxy. Particularly preferred are also compounds of formula I in which the hexose part is derived from D-glucose, n stands for 0 or 1, r\ R^ and R^ stand for C„_,-alkanoyl, R<2>stands for C, .- 3 5 7 o alkyl or phenyl, R , R and R stand independently of each other 8 o 9

for hydrogen or methyl, R stands for C^_^-alkyl, R for amino, lower alkoxy, pivaloyloxymethoxy, diphenylmethoxy, benzyloxy or 2-trimethylammonio-ethoxy, R stands for hydrogen or lower alkoxycarbonyl, R"^ stands for C^_^- alkyl, lower alkanoyloxymethyl or (2-benzyloxycarbonyl-

amino-ethyl)-sulfony1-methyl and R 12 stands for amino, lower alkoxy, pivaloyloxymethoxy, diphenylmethoxy, benzyloxy, 2-trimethylaminoethoxy, 3-cholesteryloxy or benzoyloxymethoxy, on the condition that at least one of the residues R<g>

12

and R is different from amino and lower alkoxy, and salts of such compounds which are capable of salt formation.

Particularly preferred are, above all, compounds of

9 12

formula I in which at least one of the radicals R and R represents pivaloyloxymethoxy, diphenylmethoxy, benzyloxy, 2-trimethylammonioethoxy, 3-cholesteryloxy or benzoyloxymethoxy

9 12

and the other of the residues R and R has the above meaning, and salts of such compounds which are capable of salt formation.

Most preferred are the compounds of formula I mentioned in the examples.

The compounds of formula I can be prepared in a manner known per se:

They are produced e.g. in that one

a) reacts a compound of formula II,

i. • • x. x. nla r^2a _4a _6a « ,.

in which at least one of the residues R , R , R and R stand for

hydrogen, and the other residues have the meaning of R"*",

2 4 6

the group R -C(=0)-, R respectively R , and the rest of the substituents have the meanings indicated above, whereby in a

compound of formula II present free, functional groups with the exception of those groups which are to participate in the reaction, if necessary protected with easily removable protective groups, with a transferring acylating agent for the residue r\ R<2->C(=0)-, R ^ or R^, and if necessary cleaves off protecting groups present, or

b) reacts a compound of formula

in which the substituents have the above meanings, or a reactive derivative thereof, with a compound of formula IV,

where X stands for a reactive, . esterified hydroxy group, and the other substituents have the meanings indicated above, whereby in a compound of formula IV present, free, functional groups with the exception of X are, if necessary, protected with easily cleavable groups, and if necessary then cleave off the protective groups present, or

c) reacts a compound of formula V,

in which q, r, s and t independently of each other stand for 0 or 1 and where the substituents have the meanings indicated above, whereby in a compound of formula V present, free, functional groups with the exception of the groups that 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,

where u, v and x independently stand for 0 or 1

and the other symbols and substituents have the meanings given above, in which a compound of formula VI present, free, functional groups, with the exception of the groups which are to participate in the reaction, can, if necessary, be protected with easily removable protective groups, whereby u, v and x stands for 1, when q and t in the reaction partner of the formula V stand for 0, or u stands for 0 and v and x stands 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. (for the preparation of compounds of formula I, in which n stands for 1), u and x stand for 0, when q, r, s and t stand for 1, or with a reactive derivative thereof, and then if necessary cleaves protecting 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

hydroxy and amino, and/or R"^ stands for lower alkoxycarbonyl or aryllower oxycarbonyl and the other substituents have the meanings indicated above, esterifies a compound of formula VII,

in which at least one of the residues R"^a and R"^ stands for carboxy and the other of the residues R"*"^a and R"^ has the above indicated 10.9 meaning of R , respectively of the group R -C(= 0)-, 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 the groups which will participate in the reaction, are, if necessary, protected with easily removable protective groups, or a reactive carboxylic acid derivative thereof, and if necessary cleaves off protective groups present, or e) in a compound of formula I, in which at least one of the residues R8, C(=0)-R9, R10, R11 and C(=0)-R12 is present in protected form, which does not correspond to the definition of the desired end-product, cleaves off the relevant protective groups, or f) for the production of a compound of formula I, in which R 9 stands for amino and the other substituents have the meanings indicated above, amidates a compound of formula VIII,

14

in which the residue R stands for carboxy and the other substituents have the above-mentioned meanings, whereby in a compound of formula VIII present, free, functional groups with the exception of the groups which are to participate in the reaction, can, if necessary, be protected with easily removable protective groups, or a reactive carboxylic acid derivative thereof, and if necessary cleaves off protecting groups present,

and, if desired, by carrying out one of the methods a-f) 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 divides a prepared mixture of isomers.

Methods a, c, d and e are preferred, further method f.

The implementation of the above-mentioned method variants is described in more detail in the following:

Procedure a:

Method a) is preferably used for the introduction of

14 6 o

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

In a compound of formula II, optionally present free functional groups which are preferably protected by means of easily cleavable protecting groups, in particular free hydroxy or mercapto in the residue R, are free carboxy

9 10 12

R -C(=0)-, R or R -C(=0)- as well as free amino, hydroxy or guanidino in residue R"*"^. Optional is the protection of free carbamoyl R<9->C(=0)- or R<12->C(=0)-.

Protective groups, their introduction and cleavage are described, for example, 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 are

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 may 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 diphenylmethoxycarbonyl1, or 2-halo-lower oxycarbonyl, such as 2,2,2-trichloroethoxycarbonyl, further trityl or formyl, or organic silyl or stannyl residues, further easily cleavable etherified groups, such as tert.-lower alkyl , e.g. tert.-butyl, 2-oxa- or 2-thia-aliphatic or cycloaliphatic hydrocarbon residues, primarily 1-lower-alkyllower-alkyl 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 analogues, as well as optionally substituted 1-phenyl lower alkyl, such as optionally substituted benzyl or diphenylmethyl, whereby as substituents on the phenyl residue e.g. halogen, such as chlorine, lower alkoxy, such as methoxy and/or nitro are mentioned.

Carboxyl groups are usually protected in esterified form, whereby such ester groupings are slightly 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 i.a. tert-lower oxycarbonyl, e.g. tert-butyloxycarbonyl, arylmethoxycarbonyl with 1 or 2 aryl residues, whereby these possibly constitute 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 poly-substituted 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 methoxymethoxycarbonyl, 1-methoxyethoxycarbonyl or 1-ethoxymethoxycarbonyl, 1-lower alkylthio-lower oxycarbonyl, such as 1-methylthiomethoxycarbonyl or 1-ethylthiomethoxycarbonyl, aroylmethoxycarbonyl, in which the aroyl group possibly amount to e.g. with halogen, such as bromine, substituted benzoyl, e.g. phenazyloxycarbonyl, 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-butylmethylsilyl)ethoxycarbonyl, or 2-triarylsilylethoxycarbonyl, such as 2-triphenylsilylethoxycarbonyl.

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

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

A protected amino group can e.g. available 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 alkanecarboxylic acid or optionally, e.g. with halogen, lower alkoxy or nitro, substituted benzoic acid, or 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, possibly e.g. with halogen, lower alkoxy or nitro substituted benzoyl, e.g. benzoyl, 4-chloro-benzoyl, 4-methoxybenzoyl or 4-nitrobenzoyl, or in the 1-position on the lower alkanyl radical branched or in the 1- or 2-position suitably substituted lower alkoxycarbonyl, especially tert.-lower oxycarbonyl, e.g. tert.-butyloxycarbonyl, arylmethoxycarbonyl 1 with 1 or 2 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 poly-substituted 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 optionally constitutes e.g.

with halogen, such as bromine, substituted benzoyl, e.g. phenazyloxycarbonyl, 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 represent an optionally substituted, e.g. with lower alkyl, lower alkoxy, aryl, halogen or nitro substituted aliphatic, araliphatic, cycloaliphatic or aromatic hydrocarbon residue with up to 15 C atoms, as corresponding, 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 acyl residues which come into consideration as amino protecting groups are also corresponding residues of organic phospho-, 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 phenyloxy-phenyl-phosphoryl, e.g. phenyloxyphenyl-phosphoryl, 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 aryl 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 with such a residue is primarily arylthio or aryllower alkylthio, in which aryl in particular stands for possibly 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. Such an amino protecting group is e.g. 4-nitrophenylthio.

In a 2-acyl-lower alk-l-en-l-yl residue usable as an amino protecting group, acyl is e.g. the corresponding residue from a lower alkane carboxylic 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 half-ester, such as a carboxylic acid-lower alkyl half-ester. Relevant protecting groups are primarily l-lower alkanoyl-prop-l-en-2-

howl, 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; as corresponding anions, primarily anions from strong inorganic acids, such as halogen hydrogen acids, e.g. the chlorine or bromine anion, or from organic sulphonic acids, such as p-toluenesulphonic acid in question.

Preferred amino protecting groups are acyl residues of carboxylic acid half-esters, especially tert-butyloxycarbonyl, possibly e.g. as above, 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 methoxy, substituted diphenylmethyl, such as 4,4<1->dimethoxy-diphenyl-methyl, triphenylmethyl, trimethylsilyl, benzyl-thiomethyl, tetrahydropyranyl, acylaminomethyl, benzoyl, benzyloxycarbonyl or aminocarbonyl, such as ethylaminocarbonyl.

Primary carboxylic acid amide groups (carbamoyl groups, -CONI^) are e.g. protected in the form of N-(9-xanthenyl) derivatives or in the form of N-(mono, di- or tri-arylmethyl) derivatives, whereby aryl in particular stands for unsubstituted or with up to 5 identical or different substituents substituted phenyl. Such phenyl substituents are preferably lower alkyl, such as methyl, or lower alkoxy, such as methoxy. As examples of such arylmethyl protecting groups can be mentioned: 4-methoxy-benzyl, 2,4,6-trimethoxy-benzyl, diphenyl-methyl, Di-(4'-methoxy-phenyl)-methyl and di-(4-methylphenyl) -methyl.

The protection of the carbamoyl groups is optional, i.e. under suitable reaction conditions, e.g. by using suitable condensation agents, not necessarily required. Guanidino groups are e.g. protected in the form of acid addition salts, in particular as hydrogen chloride or as toluene sulphonate.

An acyl extender which transfers the residue R 1 , R 2 -C(=0)-,

R 4 or R 6 is in particular the corresponding carboxylic acid,

i.e. R<1->OH, R<2->COOH, R<4->OH or R<6->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.

The invention relates in particular to the embodiments of the method a) whereby an acyl residue R1 R4 and/or R1 is introduced.

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 particularly esters which are unsaturated on the linking carbon atom in the esterifying residue, 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 for activated vinyl ester), carbamoyl vinyl ester (which can, for example, be obtained by treating the corresponding acid with an isoxazolium reagent; 1,2-oxazolium or Woodward method), or 1-lower oxyvinyl ester (which can be obtained, for example, by treating the corresponding acid with a lower alkoxyacetylene; ethoxyacetylene method) or ester of the amidino type , as N,N<1->disubstituted amidino ester (which can, for example, be obtained by treating the corresponding acid with a suitable N,N<1->disubstituted carbodiimide, e.g. N,N'-dicyclohexylcarbodiimide; carbodiimide method), or N,N-disubstituted amidino ester (which can be obtained, for example, by treating the corresponding acid with an N,N<1->disubstituted cyanamide; the cyanamide method), suitable aryl ester, especially by electron -attracting substituents

suitable substituted phenyl ester (which can, for example, be obtained by treating the corresponding acid with a suitable substituted phenol, e.g. 4-nitrophenol, 4-methylsulfonyl-phenol, 2,4,5-trichlorophenol, 2,3, 4,5,6-pentachlorophenol or 4-phenyl-diazophenol, in the presence of a condensing agent, such as N,N'-dicyclohexylcarbodiimide; the activated aryl ester method), cyanomethyl ester, which one e.g. can be obtained by treating the corresponding acid with chloroacetonitrile in the presence of a base; the cyanomethyl ester method), thioester, especially possibly 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, e.g. by means of the anhydride or carbodiimide method; the method with activated thiol ester), amino or amido ester, which one e.g. can be achieved by processing the equivalent

the acid, with an N-hydroxy-amino or N-hydroxy-amido compound, e.g. N-hydroxy-succinimide, N-hydroxy-piperidine, N-hydroxy-phthalimide or 1-hydroxy-benztriazole, e.g. by the anhydride or carbodiimide method; the method with activated N-hydroxy ester) or silyl ester), which one e.g.

can be obtained by treating the corresponding acid with a silylating agent, e.g. hexamethylsilazane, and which readily reacts with the hydroxy but not with the amino group).

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 acid with thionyl chloride, phosphorus pentachloride or oxalyl chloride; the acid chloride method), acids (which can be obtained, for example, from the corresponding acid ester over the corresponding hydrazide and treating this with nitric acid; the acid method), anhydrides with carbonic acid half-derivatives, as with corresponding esters, e.g. carboxylic acid lower alkyl semi-esters (which can, for example, be obtained by treating the corresponding acid with halogen, such as chloroformic acid lower alkyl esters or with a l-lower alkylcarbonyl-2-lower alkyloxy-1,2-dihydro-quinoline, e.g. l -lower oxycarbonyl-2-ethoxy-1,2-dihydroquinoline; the method with mixed O-alkyl carboxylic anhydrides), or anhydrides with dihalogenated, especially dichlorinated phosphoric acids (which can be obtained, for example, by treating the corresponding acid with phosphorus oxychloride; phosphorus oxychloride -method), or anhydrides with organic acids, such as mixed anhydrides with organic carboxylic acids (which can e.g. be obtained by treating the corresponding acid 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 one e.g. can be obtained by treating a salt, such as an alkali metal salt, of the corresponding acid, with a suitable organic sulfonic acid halide, such as lower alkane or alkyl, e.g. methane or p-toluenesulfonic acid chloride; the method with mixed sulphonic acid anhydrides), as well as symmetrical anhydrides (which can, for example, be obtained by condensation of the corresponding acid in the presence of a carbodiimide or by 1-diethylaminopropyne; the method with symmetrical anhydrides).

Suitable cyclic amides are especially amides with a 5-membered diaza-cyclic aromatic character, such as amides with imidazoles, e.g. imidazole (which can, for example, be obtained by treating the corresponding acid with N,N<1->carbonyldiimidazole; the imidazolide method), or pyrazoles, e.g. 3,5-dimethylpyrazole (which can, for example, be obtained via the acid hydrazide by treatment with acetylacetone; the pyrazolid method).

As mentioned, derivatives of acids, which are used as acylating agents, can also be formed in situ. This is how you can e.g. form N,N'-disubstituted amido ester in situ by reacting the mixture of the starting material of formula II and the acid used as acylating agent in the presence of a suitable N,N'-disubstituted carbodiimide, e.g. N,N<1->dicyclohexylcarbodiimide. Furthermore, amino or amido esters of the acids used as acylating agent can be formed in the presence of the starting material of formula II to be acylated, by reacting the mixture of the corresponding acid and amino starting materials in the presence of an N,N<1 ->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-dimethylamino-pyridine.

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 on the acylating agent is activated, usually in the presence of a suitable solvent or diluent or a mixture thereof, and, if necessary, in the presence of a condensation agent, such as also

can be an acid-binding agent, when the carboxyl group participating in the reaction is present as an anhydride, during cooling or heating, e.g. in a temperature range from approx. -30°C to approx. +150°C, especially from approx. 0 to 100°C, preferably from room temperature (approx. 20°C) to +70°C, in a closed reaction vessel and/or in an inert gas atmosphere, e.g. nitrogen. Common condensation agents are e.g. carbodiimides, e.g. N,N'-diethyl-, N,N<1->dipropyl-, N,N'-dicyclohexyl- or N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide, suitable carbonyl compounds, for example carbonyl-diimidazole, or 1,2-oxazolium compounds, e.g. 2-ethyl-5-phenyl-1,2-oxazolium-3'-sulphonate 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 sulfate), or organic bases, which are 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 are not a component of the desired end product of formula I are carried out in a manner known per se, e.g. by means of solvolysis, especially hydrolysis, 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 alkyl carbonyl 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 also transfer suitable substituted benzyloxycarbonyl, such as 4-nitrobenzyloxycarbonyl, by means of chemical reduction, e.g. by treatment with an alkali metal, e.g. sodium dithionite, or with a reducing metal, 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 evolve nascent hydrogen,

as an acid, primarily a suitable carboxylic acid, as an optional, e.g. with hydroxy, substituted lower alkanecarboxylic acid, e.g. acetic acid, formic acid, glycolic acid, diphenylglycolic acid, lactic acid, mandelic acid, 4-chloromandelic acid or tartaric acid, or an alcohol or thiol, whereby water is preferably added, to free carboxyl. By treatment with a reducing metal or metal salt, as described above, one can also convert the 2-halogen-lower oxycarbonyl (possibly after conversion of a 2-bromo-lower oxycarbonyl group to the corresponding 2-iodo-lower alkoxycarbonyl group), or aroylmethoxycarbonyl to free carboxyl , whereby aroylmethoxycarbonyl optionally

can be cleaved by treatment with a nucleophilic, preferably salt-forming reagent, such as sodium thiophenolate or sodium iodide. Substituted 2-silylethoxycarbonyl can also be transferred by treatment with a salt of hydrofluoric acid which releases a fluorine 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. pre-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 protection group in various ways, preferably by means of solvolysis or reduction. 2-halogen-lower oxycarbonylamino (optionally after conversion of a 2-bromo-lower oxycarbonylamino group to a 2-iodo-lower oxycarbonylamino group), aroylmethoxycarbonylamino or 4-nitrobenzyloxycarbonylamino can e.g. is split by treatment with a suitable chemical reducing agent, such as zinc, in the presence of a suitable carboxylic acid, such as aqueous acetic acid. Aroylmethoxycarbonylamino can also be cleaved by treatment with a nucleophilic, preferably salt-forming reagent, such as sodium thiophenolate, and 4-nitro-benzyloxycarbonylamino also by treatment with an alkali metal, e.g. sodium dithionite. Optionally substituted diphenylmethoxycarbonylamino, tert.-lower oxycarbonylamino or 2-tri-substituted 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. by treatment with an acid, such as mineral acid, e.g. hydrochloric acid, or an organic acid, 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 formed 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 emits fluoride anions, as described above in connection with the release of a corresponding protected carboxyl group.

In the form of an azido group protected amino is transferred, e.g. by reduction to free amino, 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 protected hydroxy or mercapto group 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 set free by catalytic hydrogenation, e.g.

in the presence of a palladium-on-charcoal catalyst. A with 2,2-dichloroacetyl protected hydroxy- or mercapto-

group, is set free e.g. by basic hydrolysis, an etherified hydroxy or mercapto group with a tert.-lower alkyl or with a 2-oxa- or 2-thia-aliphatic or -cycloaliphatic hydrocarbon residue is set free by azidolysis, e.g. by treatment with a mineral acid or a strong carboxylic acid, e.g. trifluoroacetic acid. Two hydroxy groups, which together are removed by means of a preferably substituted methylene group, as with 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. 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 protective groups are selected in the presence of several protected, functional groups so that more than one such group can be cleaved at the same time, for example azidolytically as in treatment with trifluoroacetic acid or formic acid, or reductively, as in treatment with zinc and acetic acid, or with hydrogen and a hydrogenation catalyst, as a palladium-charcoal catalyst. When the desired end products contain protecting groups, e.g. when R<12> stands for arylmethoxy, such as benzyloxy, the protective groups which are to be removed after completion of the reaction are selected, so that they can be removed regioselectively, e.g. can one with an organic

8 11

silyl residue etherified hydroxy in the residue R or R is set free with fluoride, whereby an arylmethoxy protecting group

9 12

remains in the remainder R or R

The starting substances of formula II in which R<2a> stands for hydrogen, can e.g. are prepared from corresponding compounds, in which R<2a> stands for an amino protecting group, e.g. benzyloxycarbonyl.

Procedure b:

A reactive derivative of a compound of formula III is e.g. a metaloxy compound such as can be obtained by reacting a compound of formula III with a suitable base,

as an alkali metal hydride or amide.

Reactive esterified hydroxy X is e.g. with a strong inorganic or organic esterified hydroxy, such as a mineral acid, e.g. hydrohalic acid, such as hydrochloric acid, hydrobromic acid or hydroiodic acid, further sulfuric acid or halosulfuric acid, e.g. fluorosulphuric acid, or with a strong organic sulphonic acid, as a possibly e.g. with halogen, such as fluorine, substituted lower alkanesulphonic 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, such as a methanesulfonic acid, trifluoromethanesulfonic acid 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 with easily cleavable protecting groups, especially hydroxy, mercapto or carboxy groups.

The reaction is preferably carried out in an inert organic solvent 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 that do not form a component of the desired final substance of formula I takes place as described under method a).

Procedure c:

A reactive carboxylic acid derivative of a compound of formula V is primarily a reactive ester, a reactive anhydride or a reactive cyclic amide, in which the carboxyl group is activated in an analogous manner 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 compound of formulas V or VI, which are preferably protected with easily removable protective groups, are in particular amino and guanidino in the residue R11, carboxy as residue R and hydroxy as residue R 9 or R<12>, further also hydroxy or mercapto

8 11

in residue R or hydroxy in residue R

In a reactive derivative of a compound of formula VI, the amino group is e.g. activated by reaction with a phosphite, e.g. diethyl chlorophosphite, 1,1-phenylene chlorophosphite, ethyl dichlorophosphite, ethylene chlorophosphite or tetraethylpyrophosphite, or by bonding to halocarbonyl, 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 group participating in the reaction is present in free form.

The reaction and the subsequent cleavage of the protecting group groups, which do not form a component of the desired end product, are carried out as described in method a).

The esterification of a compound of formula V, wherein q, r

and t stands for 1, can also be carried out using the esterification agent mentioned in method d).

Progress d:

Any free functional groups present in a compound of formula VII, which are protected with slightly cleavable protecting groups, are in particular carboxyl groups which must not be esterified, and further hydroxy, mercapto, guanidino and amino groups. Suitable protecting groups and their removal are described in method a), A reactive carboxylic acid derivative of a compound of formula VII is in particular a reactive ester, a reactive anhydride or a reactive cyclic amide, in which the carboxyl group is activated in a similar way as in those in method a) mentioned reactive acylating agents, 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 of a compound of formula VII is converted into esterification

13 9a 9a

of carboxy R with a compound H-R , in which R has the meanings given above for R 9 with the exception of hydroxy and amino, or, for esterification of carboxy R^^<a>, with a lower alkanol or aryl lower alkanol.

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

Suitable agents for esterification are, for example, corresponding diazo compounds, such as optionally substituted diazolavere-alkanes, e.g. diazomethane, diazoethane, diazo-n-butane or diphenyldiazomethane. These reagents are employed 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 halo-sulfuric acid, e.g. fluorosulphuric acid, or strong organic sulphonic acids, which may e.g. with hydroxy, 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 i.a. 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 of an ether, 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 sulphate), or organic bases, usually sterically hindered tri-lower 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 under cooling, at room temperature or under heating, e.g. at temperatures of 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 atmosphere.

Further agents for esterification are corresponding trisubstituted oxonium salts (so-called "Meerwein" salts), or disubstituted carbenzium or halonium salts, in which the substituents are the etheric residues, for example trilavereal alkyloxonium salts, as well as dilavereal oxycarbenium or dilavereal alkylhalonium salts, especially the corresponding salts with complex , 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 dimethylbrommonium 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 in a mixture thereof, if necessary, in the presence of a base, such as an organic base, e.g. a, preferably partially hindered, trilower alkyl amine, e.g. N,N-diisopropyl-N-ethylamine, and under cooling, at room temperature or under gentle 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, e.g. a nitrogen atmosphere.

A preferred embodiment of method d) is the preparation of a cesium salt of a compound of formula VII

with the desired alcohol as esterification component, in which the hydroxyl group is present in reactive esterified form.

Procedure e:

A compound of formula I, wherein at least one of the radicals R g,

R<9>, R"^, R^ and R^<2> are in protected form, which does not correspond to the definition for the desired end product, is in particular a compound of formula I in which a hydroxy or mercapto group in the residue R g, a amino or hydroxy group in the residue R 11 , free hydroxy R 9 or R 12 , and/or carboxy R is protected with an easily cleavable protective group, which is not contained in the desired end product.

Easily cleavable protecting groups are especially those which

is mentioned in procedure a). The removal of the protecting groups is carried out as described in the procedure

a) .

Procedure f:

Possibly present free, functional groups i

a compound of formula VIII, which is protected by easily removable protective groups, in particular carboxyl groups which must not be amidated, and further hydroxy, mercapto, guanidino and amino groups.

Suitable protecting groups and their removal are described by method a).

A reactive carboxylic acid derivative of a compound of formula VIII is in particular 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 carry out the reaction in such a way that a reactive carboxylic acid derivative of a compound of formula VIII is reacted with ammonia. Alternatively, you can also react a compound of formula VIII with free carboxyl groups with a reactive ammonia derivative, whereby the activation of the amino group e.g. can be carried out as described in procedure c).

The reaction and the subsequent removal of the protecting group, which is not a component of the desired end product, is carried out as described by method

a) .

Additional operations:

Salts of the compound of formula I with salt-forming groups can be prepared in a manner known per se. Consequently, one can form salts of the compounds of formula I with acidic groups by the 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 alpha-ethyl-caproic acid, or with suitable inorganic alkali or alkaline earth metal salts, especially salts derived from a weak or preferably volatile acid, e.g. sodium bicarbonate, 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 the compounds of formula I, such as e.g. contains a free carboxyl group and 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 by treatment with liquid ion exchangers.

Salts can be transferred in the usual way 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 separated in a manner known per se, e.g. by fractional crystallization, chromatography, etc., to the individual isomers.

The methods mentioned above, including the methods for cleaving off the protecting groups and the additional method precautions, are 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, in a closed container and/or in an atmosphere of inert gas, e.g. a nitrogen atmosphere.

Thereby, taking into account all substituents present in the molecule, if necessary, e.g. in the presence of easily hydrolyzable residues, particularly gentle reaction conditions are used, such as short reaction times, use of mild acids or basic agents in low concentrations, stoichiometric quantity ratios, selection of suitable catalysts, solvents, temperature and/or pressure conditions.

The invention also relates to those embodiments of the process whereby one starts from the compound obtained as an intermediate at any stage of the process and carries out the remaining process steps, or interrupts the process at any stage or uses a starting material under the reaction conditions or 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 indicated above as particularly valuable.

New starting materials and/or intermediate products as well as methods for their production are also the subject of the present invention. Such starting materials are preferably used and the reaction conditions are chosen in such a way that compounds are provided which are indicated in the present description as particularly valuable.

The invention also relates to pharmaceutical preparations which contain an effective amount, in particular an effective amount for the prophylaxis or therapy of viral infections or an effective amount for the therapy of tumor diseases, of the active substance together with pharmaceutically usable carrier substances, which are suitable for topical use, e.g. 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. diatomaceous earth, 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. starches, 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 excipients, e.g. preservatives, stabilisers, wetting agents and/or emulsifiers, solubility mediators, salts for regulating the osmotic pressure and/or buffers. Present pharmaceutical preparations which, if desired, can contain other pharmacologically active substances, such as antibiotics, are prepared in a manner known per se, e.g.

using conventional mixing, granulating, coating, dissolving or lyophilizing methods,

and contains from approx. 0.001% to 20%, especially from approx. 0.01% to approx. 10%, primarily between 0.1% and 5% of the active substance(s), whereby a concentration of active substance of less than 1% is particularly suitable for preparations for topical application.

The following are preferred as topically applicable administration forms: 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 use or lipstick, or aqueous solutions with an active substance content of 0.001% to 1%, especially 0.05% to 0.5%, preferably isotonic, sterile and physiologically tolerable solutions, e.g. eye drops, preferably in micro containers for single use, or spray forms 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. As an oily base, fatty alcohols are primarily used, 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 bisox, 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 foil alcohols or ethylene oxide addition products thereof, such as polyglycerol fatty acid esters or polyoxyethylene sorbitan fatty acid esters (Tweens), further polyoxyethylene fatty alcohol ether or fatty acid ester, 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. fetyl alcohol or stearyl alcohol. Additives to the water phases are i.a. agents which prevent drying 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. As a fatty phase, primarily hydrocarbons, e.g. petroleum jelly, paraffin oil and/or hard paraffin, which to improve the water-binding capacity preferably contains suitable hydroxy compounds, such as fatty alcohols or esters thereof, e.g. cetyl alcohol or wool wax alcohol, according to 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. glycerin, propylene glycol, sorbitol and/or polyethylene glycol, as preservatives, fragrances, etc.

Fatty ointments are anhydrous and contain hydrocarbons in particular as a basis, e.g. paraffin, vaseline and/or liquid paraffins, further natural or partly synthetic fatty substances, e.g. coconut fatty acid triglyceride, or preferably hardened oils, e.g. hydrogenated peanut, or castor oil, further fatty acid partial esters of glycerin, e.g. glycerine mono- or distearate, as well as e.g. the fatty alcohols mentioned in connection with the ointments which increase the ability to absorb hydrogen, emulsifiers and/or other additives.

Pastes are creams and ointments with secretion-absorbing powder components, 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.

Foams are administered from pressurized containers and are aerosol form present liquid oil-in-water emulsions, whereby halo-generated hydrocarbons, such as chlorine, fluorine, lower alkanes, e.g. dichlorodifluoromethane and dichlorotetrafluoroethane are used as propellants. As an oil phase, e.g. hydrocarbons, e.g. castor oil, fatty alcohols, e.g. cetyl alcohol, fatty acid esters, e.g. isopropyl meristate, and/or other waxes. As emulsifiers, e.g. mixtures of these with predominantly hydrophilic properties, such as poly-oxyethylene 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 food preservatives etc.

Tinctures and solutions mostly have an aqueous-ethanolic basis, which i.a. has added polyalcohols, e.g. glycerin, glycols, and/or polyethylene glycol, as an agent that retains moisture to reduce evaporation, and relubricating substances, such as fatty acid esters with lower polyethylene glycols, i.e. soluble in aqueous mixtures, hypophilic substances, as a replacement for the fatty substances extracted from the skin with the ethanol, and, if necessary, other auxiliaries and additives.

The preparation of the topically applicable pharmaceutical preparations is carried out in a manner known per se, e.g. by dissolving or suspending the active substance in the base, or if necessary, in a part of it. When processing the active substance as a solution, this is usually dissolved before emulsification in one of the two phases; when processing as a su suspension, it is added after emulation with part of the base and then the rest of the composition is added.

The following examples illustrate the present invention. The R^ values are measured on kieselgel thin-layer plates (company Merck, Darmstadt, Germany). The ratio of the eluents

in the eluent mixtures used are given in volume proportions (V/V), the temperatures are given in °C. The concentration, c, of the substance in the solvent (mixture) is in the case of optical rotation indicated in % (weight/volume).

Abbreviations:

abs. = absolutely

Boe = tert.butyloxycarbonyl

HV = high vacuum

i.vak. = in vacuum

Me = methyl

MeOH = methanol

PTFE = polytetrafluoroethylene, "Teflon"

RT = room temperature

Temp. = melting point

Dec. = decomposition

Example 1.

11.4 g (16.77 mmol) of N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester (a,B mixture), which contains 1.17 mol of water, is 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 stirred 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 is obtained -D-isoglutamine-benzhydryl ester (a, B - mixture) in the form of colorless crystals with m.p. 165-166°C.

The mother liquors from the recrystallization are collected and evaporated

in vacuum. The residue thus obtained 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 is obtained chromatographically pure 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester (a,B mixture), which together with the first fraction is dissolved in 200 ml of absolute methanol. The slightly cloudy solution obtained is then filtered through a millipore filter ("Fluoropore", PTFE, 0.2 µm) and the clear filter is evaporated in vacuo at 40°C. The residue is then dissolved in 50 ml of absolute acetic acid ethyl ester, which has previously been filtered through a Millipore filter ("Fluoropore", PTFE, 0.2

Hm), and is 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. 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-L-isoglutamine-benzhydryl ester (a,3 mixture) is obtained in the form of colorless crystals with m.p. 165-166°C, which still contains 0.27 mol of water.

C38H48<N>4°14"0.27 H2° <<'789 '68)

Calculated C 57.80 H 6.22 N 7.10 0 28.91 H20 0.62

Found C 57.91 H 6.20 N 7.11 0 28.77 H„0 0.62

--2 0

/a/D = +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 fraction 105-140 from the column chromatography mentioned above.

It after evaporation in vacuo obtained the yellow residue (0.55

g, foam) is dissolved in a warm state in cyclohexane-diethylether-methylene chloride (10:30:5) and the resulting solution is cooled to room temperature. Thereby a yellow oil separates which, after decanting off the solvent, is stirred for 1/2 hour with 50 ml of absolute diethyl ether. The cross numbers obtained are filtered off and washed with absolute diethyl ether. a..-.

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.

One obtains la,4,6,-tri-0-acetyl-2-deoxy-2-propionylamino-D-mannos-3-O-y1-acetyl-L-alanyl-D-isoglutamine benzhydryl ester as colorless powder, which contains 1.11 moles of water.

<C>38<H>48<N>4°14'<1>,11H2°(804'81)

Calculated C 56.71 H 6.31 N 6.96 0 30.03 H20 2.48

Found C 56.94 H 6.36 N 7.22 0 30.15 H-0 2.48

— 20

/a/^ = +5.5 + 2.7° (c=0.365; methanol) Rf=0.49 (chloroform:methanol = 9:1), Rf= 0.79 (chloroform:methanol=4:1) .

The starting material is obtained in the following way:

Step 1.1: 16.0 g (31.63 mmol) of N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine, which contains 0.74 mol of water, is dissolved in 300 ml of methanol-dimethoxyethane (1:1) and is 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, 4.6 g (23.7 mmol) of diphenyldiazamethane are added each time. After completion of the reaction (90 hours), the red solution is evaporated in vacuo at 40°C to dryness, the red resin thus 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-propionyldesmethylmuramyl-L-alanine-D-isoglutamine-benzhydryl ester is obtained in the form of colorless crystals with m.p. 188-190°C (dec.), which contains 1.117 mol of water.

C32H42<N>4°11* 1.17 H2° (679'78)

Calculated C 56.55 H 6.60 N 8.24 0 28.63 H20 3.09

Found C 56.67 H 6.67 N 8.43 0 28.65 H~0 3.09

A/q - +7.5 + 0.1° (c=0.898; methanol), Rf=0.62 (chloroform; methanol: water'(70 : 30 : 5 ) , R^=0.62 ( chloroph orm:methanol=7 , 3 ) .

Example 2.

2.33 g (3.8 mmol) of N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzyl ester which still contains 1.59 mol of water is dissolved in 24 ml of absolute pyridine, then 1.4 ml (14.8 mmol) of acetic anhydride and the resulting solution is stirred for 16 hours at room temperature. The colorless solution is then evaporated in high vacuum at 30°. You get a light yellow foam that is cleaned (10 ml fractions)

by column chromatography on 400 g of silica gel ("Type 60", pure, Merck; 0.063-2 mm) in the system chloroform-ethanol (9:1), whereby successively the a-ammoners, the a,8-anomer mixture and the Q-anomer of 1 ,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-

The L-alanyl-D-isoglutamine benzyl ester is eluted.

Fractions 112-126 (B anomers), 127-190 (a,B mixture)

and 191-260 (α-anomers) are all purified and evaporated in high vacuum at 30°C.

The residue from fractions 112-126 is crystallized from 60 ml of chloroform:diethyl ether (1:5). One obtains IB, 4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine benzyl ester in the form of colorless crystals with m.p. 189-190°C. The crystals are then dissolved in 100 ml of double-distilled water-tert.-butanol (1:1). The solution obtained is filtered through a Millipore filter ("Fluoropore", PTFE,

0.2 µm) and lyophilized in high vacuum. One obtains 1B,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine benzyl ester as a colorless powder, which contains 1.21

moles of water:; .

<C>32<H>44N4°14 "<1>,21H2°(730'52)

Calculated C 52.62 H 6.43 N 7.67 0 33.30 H20 2.98

Found C 52.97 H 6.27 N 7.86 0 33.24 H^O 22598 /a/D=+14.3+4.2° (c=0.237; dimethylformamide),

Rf=0.25 (chloroform:ethanol=9:1), Rf=0.67 (chloroform:ethanol=4:1), Rf=0.45 (chloroform:methanol=9:1).

The residue from fractions 127-190 is crystallized from 120 ml of chloroform-diethyl ether (1:5). One obtains 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzyl ester (a B mixture, according to "'"H-NMR a: B*6 :1) in form of colorless crystals with m.p. 156-157°C.

The crystals are then dissolved in 150 ml of double-distilled water-tert.-butanol (1:1). The solution obtained is filtered through a Millipore filter ("Fluoropore", PFTE,

0.2 µm) and lyophilized in high vacuum. 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine benzyl ester (rv, B mixture) is obtained as a colorless powder,

which contains 1.00 mol of water.

C32H44<N>4°14* 1.00 H2° t726'73)

Calculated C 52.89, H 6.41 N 7.71 O 33.02 H20 2.48 Found C 53.18 H.6.42 N 7.49 0 32.81 ?H~0.2.48

ia/=+56.0 +1.1° (c=0.948; dimethylformamide),

Rf=0.17 + 0.24 (chloroform:ethanol = 1:1; double spot), R^=0.45 (chloroform:methanol = 1:1), R^=0.67 (chloroform:methanol=4 :1=.

The residue from fraction 191-260 is crystallized from 90 ml of chloroform-diethyl ether (1:15). One obtains 1a,4,6,-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine benzyl ester in the form of colorless crystals with m.p. 147-149°.

The crystals are then dissolved in 100 ml double distilled water - tert.-butanol (1:1). The solution obtained is filtered through a Millipore filter ("Fluoropore", PTFE,

0.2 µm) and lyophilized in high vacuum. Ia,4,6-tri-O-acetyl-desmethyl-muramyl-N-propionyl-L-alanyl-D-isoglutamine benzyl ester is obtained as a colorless powder, which still contains 1.12 mol of water.

C32H44<N>4°14* 1.12 H2° <728'89)

Calculated C 52.74 H 6.43 N 7.69 O 33.18 H20 2.76

Found C 52.96 H 6.54 N 7.62 O 33.15 H_0 2.76

ia/*=+36.9 + 1.7° (c=0.593; methylene chloride:methanol=1:1), Rf=0.17 (chloroform:ethanol = 9:1), Rf = 0.45 (chloroform: methanol = 9:1), Rf = 0.67 (chloroform:methanol = 4:1).

Example 3.

Analogous to example 1, one obtains from 1.19 g (1.54 mmol) N-propionyl-desmethylmuramyl-L-alanyl-isoglutaminyl-L-alanine benzhydryl ester (a,B mixture) containing 2.24 mol of 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- benzhy dry ester (a , B mixture) as colorless powder containing 1.3 3 moles of water.

C„ nH^N.-O, c . 1.33 H_0 (879.85)

Calculated C 55.97 H 6.40 N 7.96 0 29.69 H20 2.72

Found C 56.14 H 6.36 N 7.98 0 29.56 H~0 2.72

— 20

/ a/* = +23.8 +2.1° (c=0.478; dimethylformamide), Rf=0.36 (chloroform:methane1=9:1), R^=0.69 (chloroform:methane1=4: 1), R^=0.87 (chloroform:methanol=7:3).

The starting point is obtained in the following way:

Step 3. 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-dimethoxyethane (1:1) (18 hours, room temperature), N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine benzhydryl residues as colorless powder, containing 2, 24 moles of water.

C35H47<N>5°12'2.24 H2° <770'14)

Calculated C 54.58 H 6.76 N 9.09 O 29.59 H20 5.25

Found C 54.85 H 6.86 N 9.11 0 29.21 H^O 5.25

— 20

la/* = +2.9 + 2° (c=0.478; dimethylformamide), Rf=0.57 (chloroform:methanol = 7:3), R^= 0.65 (chloroform:methanol:water = 70:30 :5).

Example 4.

Analogous to example 1, one obtains from 0.767 g (1.09 mmol) N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid-(Ca)-methyl ester-(C^)-benzhydryl ester, which still contains 1.42 mol of water , with 0.49 ml (5.2 mmol) of acetic anhydride in 8.7

ml pyridine (44 hours, room temperature) 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid-(C )-methyl ester-(Cy-benzhydryl ester (a,B mixture ) as colorless powder, if contained 0.71 mol of water: C39<H>49<N>3°150.71 H2° <812'62)

Calculated C 57.65 H 6.28 N 5.17 0 30.92 H20 1.57

Found C 58.09 H 6.23 N 5.28 0 20.71 HO 1.57

-> n

/a/p = +53.9 + 1.9° (c=0.519; dimethylformamide), Rf=0.55 (chloroform:methanol=9:1), Rf=0.87 (chloroform:methanol=4:1 ), R^ = 0.95 (chloroform/methanol = 7:3).

The starting material is obtained in the following way:

Step 4.1:

Analogously to example 1, one obtains from 1.32 g (2.5 mmol) N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid (Ca)-methyl ester, which still contains 0.68 mol of water, with a total of 1.4 g (7.2 mmol) dimethyldiazomethane in 27 ml methanol-dimethoxyethane (1:1) (18 hours, room temperature) N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid-(Cq)-methyl ester-(C^)- benzhydryl ester as colorless powder containing 1.42 moles of water.

C33<H>43<N>3012. 1.42H20(699.30.)!

Calculated C 56.69 H 6.63 N 6.01 0 30.70 H20 3.65

Found C 56.75 H 6.73 N 6.21 O 30.74 H^O 3.65

--20

/a/p = +10.3 + 2.3° (c=0.435; water), Rf= 0.73 (chloroform; methanol:water = 70:30:5), Rf = 0.91 (chloroform:methanol = 7:3).

Example 5.

Analogous to example 1, one obtains from 1.15 g (1.57 mmol) N-propiony1-desmethylmuramyl-L-alanyl-D-glutamic acid-(Ca)-n-butyl ester-()-benzhydryl ester which still contains 0.9'9 'mol vvahn. with'0.TfDml ( 7 , 4 mmol) of acetic anhydride in 12 ml of pyridine (16 hours, room temperature), 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid-( Ca)-n-butyl ester-(C^)-benzhydryl ester (a,B mixture) as a colorless powder containing 0.73 moles of water.

C42H55<N>3°15-0.73 H2° (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 O 29.49 H20 1.54

o

/a/p" = +12.6 + 2.3° (c=0.443; chloroform), Rf=0.72 (chloroform/methanol = 9:1), R^=0.96 (chloroform/methanol = 4:1).

Starting material-light is obtained in the following way:

Step 5. 1:

Analogous to example 1, one obtains from 1.9 g (3.3 mmol) of N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid-(C )-n-butyl ester, which contains 1.03 mol of water, with a total of 2, 5g (12.85 mmol) diphenyldiazomethane in 40 ml methanol-dimethoxyethane (1:1; 70 hours, room temperature) N-propionyl-desmethyl-muramyl-L-alanyl-D-glutamic acid-(Ca)-n-butyl ester-(C ^)-Benzhydryl ester as colorless powder, containing 0.99 mol of water..

C34<H>49<N>3°12'0.99 H2° <733'62)

Calculated C 58.94 H 7.03 N 5.73 0 28.32 H20 2.43

Found C 59.24 H 7.06 N 5.59 0 28.08 H~0 2.43

/a/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 = 3:3).

Example 6.

Analogously to example 1, one obtains from 1.0 g (1.43 mmol) crude N-acetyl-muramyl-L-alanyl-D-glutamic acid-{C^)-pivaloyloxymethyl-ester-()-benzyl ester and 0.57 ml ( 6.0 mmol) acetic anhydride in 10 ml absolute pyridine (20 hours, room temperature) N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-alanyl-D-glutamic acid-()-pivaloyloxymethyl ester-(C ^)-butyl ester (a,3 mixture) as colorless powder containing 0.79 mol of water.

<C>36<H>49<N>3012. 0.99 H2O (733.62)

Calculated C 54.47 H 6.60 N 5.02 0 33.96 H20 1.69

Found C 54.52 H 6.36 N 5.12 0 33.71 HO 1.69

--2 0

/a/p = +12.5 + 1.9° (c = 0.522; methanol), Rf = 0.46 (chloroform:methanol = 9:1), R^ = 0.63 (chloroform:methanol = 4: 1),

R 2 = 0.88 (chloroform:methanol = 7:3).

The starting material is obtained in the following way:

Step 6:1:

20.2 g (60 mmol) of N-tert.-butyloxycarbonyl-D-glutamic acid-(Cy)-benzyl ester are dissolved in a mixture of 300 ml of tetrahydrofuran and 50 ml of water. 19.55 g (60 mmol) of cesium carbonate (purum, Fluka) is added to this solution,

dissolved in 100 ml of water, and let it all stand for 1/2 hour at room temperature.". Then evaporate in high vacuum at 30°C

and the obtained residue is evaporated successively with 200 ml of methanol and twice with 200 ml of dimethylformamide in high vacuum at 40°C.

The obtained cesium salt of N-tert-butyloxycarbonyl-D-glutamic acid-(C )-benzyl ester (28.1 g; 60 mmol) is suspended in 500 ml of dimethylformamide. To this suspension, at room temperature, add 18.0 g (17.4 ml; 120 mmol) pivalic acid chloromethyl ester (purum, Fluka) and 18.0 g (120 mmol) sodium iodide. The resulting mixture is stirred for 17

hours at room temperature. It is then filtered and file-

the funnel is evaporated in high vacuum at 40°C. The achieved res-

is taken up in 500 ml of acetic acid ethyl ester and washed successively 3 times with 150 ml of 0.1N sodium thiosulphate solution and water. After drying the acetate phase over disodium sulphate, it is re-evaporated in a vacuum. The still slightly yellow residue is recrystallized from 450 ml methanol-water (1:2).

N-tert-butyloxycarbonyl-D-glutamic acid-(C )-pival-oyloxymethyl ester-(C )-benzyl ester is obtained in the form of colorless crystals with m.p. 76-77°C.

C23H33NOg (451.52)

Calculated C 61.18 H 7.37 N 3.10 O 28.35

Found C 61.08 H 7.25 N 3.33j 0 28.58

/a/p = + 18.4 + 0.1° (c=1.034; acetic acid ethyl ester),

R f = 0.85 (methylene chloride: methanol: 9:1).

Step 6. 2:

20.9 g (4.4, 3 mmol) of N-tert.-butyloxycarbonyl-D-glutamic acid-(C a )-pivaloyloxymethyl ester-(C )-benzyl ester are stirred at 0°C in a mixture of 400 ml of trifluoroacetic acid-methylene chloride, (1 :1) for 1.5 hours. The reaction solution is then evaporated in a high vacuum at 30°C and the residue obtained is taken up in methylene chloride and evaporated again. You get a yellow oil which is taken up in a mixture of 100 ml of water and 300

ml of diethyl ether and cool to 0°C. 80 ml IN caustic soda (pH 7.5) is added to the solution with vigorous stirring

and separates the ester phase. The ethereal solution is extracted once with 100 ml of water, the aqueous phase is extracted once with 200 ml of diethyl ether. The ether fractions are collected, dried over sodium sulfate and filtered. The reddish filtrate is then mixed with 8.43 g (44.3 mmol) of p-toluenesulfonic acid monohydrate (puriss., Fluka), dissolved in 100 ml of diethyl ether and stirred for 1 hour at room temperature, then 1/2 hour at 0°C. The crystals obtained are suctioned off and washed with diethyl ether. The crude product obtained is recrystallized from 400 ml of acetic acid ethyl ester. You get D-glutamic acid-

(C a )-pivaloyloxymethyl ester-(C )-benzyl ester tosylate in the form of colorless crystals with m.p. 135-137°C.

C25H33N09S (523.60)

Calculated C 57.35 H 6.35 N 2.68 0 27.50 S 6.12

Found C 57.39 H 6.52 N 2.72 0 27.53 S 6.12

— 20

/a/*=+8.8 + 0.1° (c=0.924; methylene chloride),

Rf = 0.78 (methylene chloride:methanol = 9:1; double spot),

Rf = 0.84 (methylene chloride:methanol = 5:1; double spot).

Step 6:3:

2.6 g (6.5 mmol) N-acetyl-muramyl-rL-alanine sodium salt,

which contains 0.63 mol of water, is dissolved in 50 ml of dimethylformamide. To this solution is added at room temperature 1.47 g (7.14 mmol) of N,N-dicyclohexylcarbodiimide,

0.74 g (7.14 mmol) of N-hydroxysuccinimide and 3.41 g (6.5 mmol) of D-glutamic acid-(C )-pivaloyloxymethyl ester-(C )-benzyl ester tosylate and stirring the resulting clear, colored -

dissolve the solution for 14 hours at room temperature.

The crystals precipitated during the reaction (N,N-dicyclohexylurea) are then suctioned off and the filtrate is evaporated in high vacuum at 40°. The residue is taken up in 300 ml of acetic acid ethyl ester, filtered, and the solution obtained is washed 4 times in succession, each time with 100 ml of water, 3 times with 100 ml of saturated aqueous sodium sulphate solution and 2 times with 100 ml of water.

The acetate fractions are collected, dried over sodium sulfate, filtered and re-evaporated.

A faint yellow foam is obtained which is purified by column chromatography on 440 g of silica gel ("type 60", pure, Merck; 0.063-0.200 mm) in the system acetone-acetic acid ethyl ester (7:3) (10 ml fractions). Fractions 20-70 are collected and evaporated in vacuum at 40°C. Slightly contaminated N-acetyl-muramyl-L-alanyl-D-glutamic acid-(C^)-pivaloyloxymethylester-(C^)-benzyl ester is obtained as a colorless foam, which is processed without further purification.

Rf = 0.74 (chloroform:methanol:water = 70:50:5), Rf = 0.85 (chloroform:methanol = 7:3).

Example 7.

10 g (15.86 mmol) of N-benzoyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzyl ester are dissolved in 100 ml of pyridine and mixed with 9 ml of acetic anhydride. After 24 hours at room temperature, it is evaporated under high vacuum, the residue is taken up in methylene chloride and the organic phase is successively extracted with 300 ml 1N hydrochloric acid, water, 5% NaHCO 3 solution and water. The aqueous phases are extracted twice with 150 ml of methylene chloride.

The combined organic phases are dried with sodium sulphate and evaporated.

For crystallization, dissolve in 200 ml of ethanol and add 30 ml of acetic acid ethyl ester and then diethyl ether until cloudiness (approximately 60 ml). Colorless crystals of α,6-1,4,6-tri-O-acety1-N-benzoyl1-desmethylmuramy1-L-alany1-D-isoglutamine benzyl esters are obtained with m.p. 168-169°C. Additional crystals can be extracted from the mother liquor.

lg./*=+44.9° + 1.2° (c = 0.813, CHC13),

Rf = 0.54 (CHCl 3 :methanol = 9:1).

Example 8.

4.5 g (6.4 mmol) of 1,4,6-tri-O-acetyl-N-benzoyl-desmethylmuramyl-L-alanyl-D-isoglutamine, dissolved in 60 ml of acetonitrile, are mixed with 3.7 g ( 19.1 mmol) of diphenyldiazomethane in 30 ml of acetonitrile. After 5 hours at room temperature, the reaction is finished; evaporated to dryness in vacuum, the residue is extracted 4 times, each time with 20 ml of diethyl ether and purified chromatographically over 250 g of silica gel (Merck>0.04-0.6 mm). Elute step by step with one liter of methylene chloride-acetone (9:1, fraction 1), per fraction 500 ml of CP^Cl^-acetone (7:3, fractions 2 and 3) and per fraction 1 liter of Cr^C^-acetone ( 1:1, fractions 4, 5 and 6). One obtains from fraction 1 impure α-anomer, from fraction 3 pure α-anomer and from fractions 4 and 5 a,B-anomer mixture of 1,4,6-tri-O-acety1-N-benzoyl1-desmethylmuramy1-L-alany1 -D-isoglutamine benzhydryl ester.

a-anomer: m.p. 108-110°,

/a/p = +36.3 + 1 (c== 1.01, CHC13), Rf = 0.27 (CHC13;acetone = 1.1)•

The starting material, 1,4,6-tri-O-acetyl-N-benzoyl-desmethyl-muramyl-L-alany1-D-isoglutamine, is obtained by catalytic hydrogenation of the corresponding benzyl ester (Example 7)

with 5% palladium charcoal in dimethoxyethane as anomer mixture. Temp. 114-120°,

--2 0

ga/p = +51.9° +1° (c = 0.963, H 2 O), Rf = 0.16 (CHCl 3 :methanol:H 2 O = 85:15:2).

Example 9.

1.5 g (2 mmol) N-benzoyl-1a, 6), 4, 6-tri-B-buty±yl-desmethyl-

muramy1-L-alanyl-D-isoglutamine is reacted analogously to example 8 with diphenyldiazomethane. After analogous processing, a crude product is obtained which is chromatographed on 50 g of silica gel (Merck, 0.04-0.6 mm). Elute step by step with 1 liter of CF^C^-acetone (9:1, fraction 1), per fraction 1 liter of CHCl^C^-acetone (8:2, fractions 2 and 3) and per fraction 500

ml of Ci^C^-acetone (7:3, fractions 4 and 5).

From fraction 2 you get -anomer, from fraction 4 impure e-anomer of N-benzoyl-1,4,6-tri-O-butyryl-1-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester. The g-anomer gets

one crystalline from solution in acetone (0.7 ml/g) and addition of diethyl ether (22 ml/g). Temp. 109-113°,

/α/2°= +45.1° + 1.1° (c 0 0.92, CHCl 3 ), Rf = 0.62 (CHC 13 : acetone - 1:1).

The B-anomer is obtained crystalline by dissolving in 8 ml of acetone at 50°C and adding 20 ml of diethyl ether at room temperature.

Temp. 169-171°, /α/<2>°= +13.8° + 1.3° (c.=_0.741, CHCl3: methanol # 1:1), Rf = 0.52 (CHC13: acetone = 1:1).

The starting material is obtained in the following way:

Step 9:1: N-benzoyl-(1a,8),4,6-tri-O-butyryl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzyl ester is obtained in a known manner from N-benzoyl-desmethylmuramyl- L-alanyl-D-isoglutamine-benzyl ester in pyridine with 6 equivalents of succinic anhydride with the addition of catalytic amounts of 4-dimethylaminopyridine. After normal work-up, crystals of m.p. are obtained by stirring with diethyl ether. 166-171°, /a/<2>°= +43.2° +1° (c = 0.954, CHC13), a-anomer: Rf = 0.42 (CHC13:acetone = 1:1), g- anomers: Rf = 0.28 (CHC13:acetone = 1:1).

Step 9, 2:

By catalytic hydrogenation of N-benzoyl-(1a,8),4,6-tri-O-butyry 1-desmethylmuramyl-L-alanyl-D-isoglutamine benzyl ester with 5% palladium-carbon in tetrahydrofuran, N-benzoyl- (la , B £, 4 , 6-tri-O-butyryl-desmethylmuramyl-L-alanyl-D-isoglutamine as hemihydrate with m.p. 100-105°,

— ? o

/a/p = +54.2° +1° (c = 0.96, methanol), Rf = 0.45 (CHCl 3 ; methanol = 8:2).

Example 10.

Analogous to example 1, 3.69 g (5.6 mmol) of N-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester, dissolved in 40 ml of absolute pyridine, are reacted with 2.04 g (20 mmol) of acetic anhydride (1 hour, room temperature). The clear solution is evaporated at 30°C, the residue is taken up in 150 ml of acetic acid ethyl ester and the solution is extracted twice, each time with 50 ml of water. After drying and evaporation of the solvent, N-acetyl-(1a,8)-4,6-tri-O-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine benzhydryl ester is obtained.

/a-/p = +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 R^ = 0.81 (ethyl acetate:n-butanol:pyridine: acetic acid : water = 421.21:21:6:10).

N-acety1-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydry1-ester can be prepared in the following way:

Step 10. 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) of 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 a solid, 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 diethyl ether: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. In this way N-acetyl-desmethylmuramy1-L-alanyl-D-isoglutamine benzhydryl ester is obtained in the form of cube-shaped crystals.

Temp. 170°C (with decomposition),

/a/D = + 14_+ 1° (c = 1.5, methanol), Rf = 0.40 (chloroform: methanol: water = 70:30:5) and Rf = 0.66 (ethyl acetate: n-butanol :pyridine:éåd&Sik:water = 42:21:21:6:10).

Example 11.

Analogously to example 10, one obtains from 0.55 g (0.85 mmol) N-acetyl-desmethylmuramyl-L-α-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-(1a,B)-4,6-tri-O-acetyl-desmethylmuramyl-L-a-amino-butyryl-D-isoglutamine- benzhydryl ester; Rf = 0.63 (chloroform?-methanol:water = 70:30:5) and Rf = 0.82 (acetic acid ethyl ether: n-butanol:pyridine:acetic acid:water = 42:21:21:6:10).

Example 12:

From 2.10 g (2.54 mmol) of N-acetyl-desmethylmuramyl-L-α-amino-butyryl-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; crown size 0.063-0.200

etc.; 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 after the addition of 40 ml of abs. dioxane through a millipore filter ("Fluorpore", PTFE, 0.2 µm)

and then lyophilized. N-acetyl-(lat}B)-4,6-tri-O-acetyl-desmethylmuramyl-L-a-aminobutyric-D-glutamic acid dibenzhydryl ester is obtained as a colorless, light powder containing 0.37 mol of water.

C^Hg-N-O,,.<0>,37 Ho0 (958.69)

Dl 3/ J lbZ

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/p = + 29.6 + 1.7° (c = 0.577; methanol), Rf = 0.69 (n-butanol:acetic acid :water = 75:7.5:21) and R f = 0.93 (acetic acid ethyl ester : n-butanol : pyridine : acetic acid : water = 42:21:21:6:10).

The starting material is obtained analogously to example 1 in the following way: 3.00 g (6.1 mmol) N-acetyl-desmethylmuramyl-L-α-aminobutyryl-D-glutamic acid, dissolved in 30 ml of a 1:1 mixture of dimethylformamide and methanol , is mixed at room temperature and with stirring with an excess of diphenyldiazomethane. After 3 days, the red suspension is evaporated in high vacuum at 30°C, the red residue is triturated several times with diethyl ether-petroleum ether (3:1) and the supernatant liquid is decanted off. 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); blue. to remove the half-ester), 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-desmethyl-muramyl-L-a-aminobutyryl-D-glutamic acid dibenzhydryl ester is obtained as a colorless powder, 0.93 mol of water.

C45H51<N>3°12"0.93 H2° t842'67)

Calculated C 64.14 H 6.34 N 4.99 H20 1.99

Found C 64.1 H6.5 N4.9 H~0 2.0

/a/p = +11.6 + 2.9° (c = 0.344; methanol), Rf = 0.55 (acetic acid-ethyl ester: methanol = 4:1) and R^ = 0.77 (chloroform: isopropanol = 1:1).

Example 13.

Analogously to example 1, one obtains from 0.67 g (1 mmol) N-acetyl-desmethylmuramyl-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,8)-4,6-tri-O-acetyl-desmethylmuramyl-L-valyl-D-isoglutamine benzhydryl ester.

/a/20= 0 35° +1° (c = 0.594; methanol), R 0.92 (chloroform : methanol : water = 70:30:5) and R^ = 0.80 (ethyl acetate :n-butanol: pyridine.acetic acid:water = 42:21:21:6:10).

The starting material is obtained analogously to example 1 in the following way:

Step 13. 1:

1.60 g (3.16 mmol) of N-acetyl-desmethylmuramyl-L-valyl-D-isoglutamine, dissolved in 30 ml of methanol, esterified with excess 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/p = +13° + 1° (c = 01.067; methanol), Rf = 0.56 (chloroform: methanol: water = 70:30:5), Rf = 0.42 (n-butanol: acetic acid:water = 75:7.5:21) and R^= 0.72 (acetic acid ethyl ester: n-butanol:pyridine:acetic acid:water = 42:21:21:6:10).

Example 14:

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 onto 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, R^= 0.3 (methylene chloride: methanol = 5: 1).

The starting material used is produced in the following way:

Step 14:1:

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-L-valyl-D-isoglutamine-benzhydryl ester melts at 185°C (dec.), Rf = ©.5 (chloroform:methanol:water - 14:6:1), /a/* 0 +38° (c = 0.912; methanol).

Example 15:

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 common salt solution, and dried over magnesium sulfate. After evaporation of the solvent, N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-alany1-D-isoglutamine benzhydryl ester is obtained. R^ = 0.7 (methylene chloride:methanol = 5:1).

The starting material used is produced as follows

manner:

Step 15. 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 for 20 hours at 40 °C. After evaporation of the solvent, the residue is thoroughly extracted with diethyl ether and water and dried over NaOH pellets. N-acetyl-muramy1-L-alanyl-D-isoglutamine-benzhydryl ester is obtained in the form of white crystals. R^ = 0.6 (methylene chloride:methanol:water = 14:6:1).

Example 16:

1.45 g (1.99 mmol) N-acetyl-muramyl-L-alanyl-D-isoblutaminyl-L-alanjm-benzhydryl ester, dissolved in 10 ml abs. pyridine, is mixed with 0.73 g (7.15 mmol) of acetic anhydride and allowed to stand for 35 hours at room temperature. The yellowish solution is evaporated in high vacuum at 30° to dryness, the residue is mixed with abs. dioxane and lyophilized. The freeze-dried material is taken up in 10 ml chloroform-methanol (1:1) and precipitated with 100 ml abs. diethyl ether (3 times). The obtained powder is dissolved in 100 ml abs. dioxane-water (1:1), filtered as usual through a millipore filter (PTFE; 0.2 (im) and lyophilized. N-acetyl-(1a,B),4 , 6-tri-O-acety1 is obtained -muramyl-L-alanyl-D-isoglutaminyl-L-alanine-benzhydry1-ester as colorless powder, containing 1.42 mol of water and 0.7 mol of dioxane.

<C>41<H>51<N>5°15<*><0>,<7><C>4<H>0°2*<1,4>2 H2°

Calculated C 55.90 H 6.37 N 7.44 H20 2.72

Found C 55.98 H 6.42 N 7.76 H90 2.71

/a-/p = +25.5 + 2.1° (c = 0.475; dimethylsulfoxide), Rf %

0.39 (n-butanol:acetic acid:water = 75:7.5:21), R f = 0.83 (chloroform:methanol:water = 70:30:5) and R^= 0.85 (chloroform: methanol:water:acetic acid = 70:40:9:1).

The starting material is obtained in the following way:

Step 16. 1:

3.25 g (5.6 mmol) of N-acetyl-muramyl-L-alanyl-D-isoglutaminyl-L-alanine in 50 ml of methanol and esterified while stirring with excess diphenyldiazomethane. After 6 hours, the violet solution is evaporated and the yellowish residue is purified by chromatography on 400 g of silica gel ("type 60", Merck) in the system chloroform:methanol:water (70:30:5) (13 ml fractions).

The material obtained after 600 ml in fractions 42 to 60 is collected, taken up in 100 ml of chloroform, filtered through a millipore filter (PTFE; 0.2 (im) and evaporated. The remainder is taken up in 150 ml of abs. dioxane and lyophilized N-acety1-muramyl-L-alanyl-D-isoglutaminy1-L-alanine benzhydryl ester is obtained as a colorless, loose powder.

C35H47<N>5°12• 1.51 H2° <756'99)

Calculated C 55.54 H 6.69 N 9.25 H20 3.59

Found C 55.68 H 6.39 N 9.04 H90 3.59

Z«/d +16'3 ± 10 (c = 0/979; dimethylsulfoxide), Rf = 0.31 (n-butanol:acetic acid:water = 70:7.5:21), R^= 0.57 ( chloros.. form:methanol:water = 70:30:5) and R^ = 0.33 (chloroform:methanol noi:water:acetic acid = 75:27:5:0.5).

Example 17:

Analogously to example 16, one obtains from 1.00 g (1.53 mmol) N-acetyl-muramyl-L-alanyl-D-isoglutaminyl-L-alanine benzyl ester, dissolved in 18 ml abs. pyridine, and 0.66 g (5.97 mmol) of the acetic anhydride peracetyl compound (6 hours, room temperature). The purification takes place by chromatography on 150 g of silica gel ("type 60", Merck) in the system chlorine-methanol-water (70:30:5; 5 ml fractions). N-acety1-(1a,8),4,6-tri-O-acetyl-muramyl-L-alanyl-D-isoglutaminyl-L-alanine benzyl ester is obtained as a colorless lyophilisate containing 1.84 mol of water.

<C>35<H>49<N>5°15<*><1>,84H2°<812'94)

Calculated C 51.72 H 6.56 N 8.62 H20 4.07

Found C 51.5 H 6.9 N 8.3 H„0 4.1

Za/p = +38.5 + 3.5° (c = 0.282; dimethylformamide), Rf=

0.63 (acetonitrile:water = 3:1), R^ = 0.39 (n-butanol: acetic acid: water = 75:7.5:21), R^ = 0.83 (ethyl acetate : n-butanol : pyridine : acetic acid : water = 42:21:21:6:10).

Example 18.

1.45 g (1.5 mmol) Nacetyl-muramyl-L-alanyl-D-isoglutaminyl-L-alanine cholesteryl-3-ester and 0.72 g (7.1 mmol) acetic anhydride are dissolved in 16 ml of a mixture of abs.pyridine and dimethylformamide (6:1) and allowed to stand for 22 hours at room temperature. The clear solution is evaporated in high vacuum at 30°C to approx. 4 ml, the gel-like residue formed is triturated with 50 ml ethyl acetate and the supernatant liquid is decanted (4 times). The solid residue is taken up in 5 ml of dimethylformamide, 50 ml of abs. dioxane is added, the whole is filtered through a millipore filter (PTFE; 0.2 µm) and

lyophilized. N-acetyl-(1a,8),4,6-tri-0-acetyl-muramy1-L-alanyl-D-isoglutaminyl-L-alanine cholestery1-3 ester is obtained as a colorless powder containing 0.62 mol water.

<C>55<H>g7<N>5<0>15.<0>,62H20(1069.49)

Calculated C 61.77 H 8.33 N 6.55 H20 1.04

Found C 61.8 H 8.4 N 6.8 H^O 1.0

/a/p = = +7.7 +1° (c = 0.977; dimethylsulfoxide), Rf =

0.41 (n-butanol:acetic acid:water = 75:7.5:21), Rf = 0.87 (acetic acid ethyl ester:n-butanol:pyridine:acetic acid:water = 42:21:21:6:10).

Example 19.

Analogous to example 1, one obtains from N-acetyl-muramyl-L-alany1-D-glutamine pivaloyloxymethyl ester and acetic anhydride

in absolute pyridine N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-alanyl-D-glutamine pivaloyloxymethyl ester.

The starting material is obtained in the following way:

Step 19. 1:

3.0 g (8.54 mmol) of N-benzyloxycarbonyl-L-alanyl-D-glutamine are dissolved in a mixture of 300 ml of tetrahydrofuran and 20 ml of water. 1.39 is added to this solution

g (4.27 mmol) cesium carbonate (purum, Fluka), dissolved in

7 ml of water, and let it all stand for 1/2 hour at room temperature. It is then evaporated in a high vacuum at 30°C, and the residue obtained is evaporated with 250 ml of methanol and twice with 250

ml of dimethylformamide in high vacuum at 40°C.

The obtained cesium salt of the N-benzyloxycarbonyl-L-alanyl-D-glutamine is suspended in 300 ml of dimethylformamide. At room temperature, 2.57 g (2.48 ml; 17.08 mmol) of pivalic acid chloromethyl ester (purum, Fluka) and 2.56 g (17.08 mmol) of sodium iodide are added to this suspension. The resulting mixture is stirred for 42 hours at room temperature. It is then evaporated in a high vacuum at 40°C. The yellow, crystalline residue obtained is taken up in 500 ml of ethyl acetate

(suspension) and washed 3 times with 150 ml of water. The clear acetate phases are collected, dried over sodium sulfate and evaporated in vacuo at 30°C. The residue is crystallized from acetic acid ethyl ester: n-pentane = 1:15 (20:300 ml). After recrystallization twice, N-benzyloxycarbonyl-L-alanyl-D-glutamine pivaloyloxymethyl ester is obtained in the form of colorless crystals with m.p. 109-110°C.

C22H31N3°8 <465'50)

Calculated C 56.76 H 6.71 N 9.03 0 27.49

Found C 56.42 H 6.69 N 8.82 0 27.73

/a/D = +5.5 + 0.1° (c = 0.973;methanol), Rf = 0.62 (methylene chloride:methanol = 5:1), R^0.86 (methylene chloride:methanol:water = 70 :30:5).

Step 19, 2:

A solution of 6.7 g (14.4 mmol) of N-benzyloxycarbonyl-L-alanyl-D-glutamine-pivaloyloxymethyl ester in 200 ml of dimethoxyethane is hydrated at a constant pH value of 5.5 (titration with IN hydrochloric acid) with 1.0 g 10% palladium charcoal as a catalyst for 40 minutes at room temperature and under normal pressure.

The catalyst is then filtered off and the filtrate is evaporated

in high vacuum at 30°C. The residue obtained is suspended in 30 ml of diethyl ether. The colorless crystals obtained are filtered off with suction and washed with diethyl ether. L-alanyl-D-glutamine pivaloyloxymethyl ester hydrochloride is obtained in the form of colorless crystals with m.p. 110-111°C (dec.).

C14H26C1N3°6 <467'83)

Calculated C 45.72 H 7.13 Cl 9.64 N 11.42 O 26.10

Found C 45.51 H 7.36 Cl 9.34 N 11.47 0 26.30

/a/p" = +25.5 0.1° (d = 1.128; methanol), Rf = 0.07 (methylene chloride: methanol =)= 5:1), Rf = 0.37 (methylene chloride: methanol: water = 70:30;5), R^ = 0.59 (methylene chloride:methanol:water = 5:5:1).

Step 19. 3:

5.18 g (13.0 mmol; 2.52 mmol/g) of N-acetyl-4-0,6-O-isopropylidene-muramic acid sodium salt are suspended in 150 ml of dimethylformamide. 4.8 g (13.0 mmol) L-alanyl-D-glutamine pivaloyloxymethyl ester hydrochloride, 2.95 g (14.3 mmol) dicyclohexylcarbodiimide and 1.64 g (14.3 mmol) are then added at room temperature N-Hydroxy-succinimide. The slightly yellow suspension obtained is stirred for 22 hours at room temperature. The formed dicyclohexylurea is then suctioned off from the main quantity and the filtrate obtained is evaporated in high vacuum at 40°C. The remaining yellow oil is taken up in 20 ml of methanol, a further fraction of dicyclohexylurea is sucked off and it is re-evaporated under high vacuum.

The crude product obtained, which still contains some dicyclohexylurea, is purified by column chromatography on 1000 g silica gel ("type 60", pure, Merck; 0.063-0.2 mm)

in the system methylene chloride:methanol = 9:1 (10 ml fractions).

Fractions 380-1210 are collected, whereby fraction 850 is eluted with the system methylene chloride:methanol:water = 70:30:5.

After evaporation of the eluate in high vacuum at 30°C, a mixture is obtained which mainly consists of N-acetyl-4-0,6-0-isopropylidene-muramyl-L-alanyl-D-glutamine-pivaloyloxymethyl-ester and some N-acetyl -muramyl-L-alanyl-D-glutamine-pivaloyl-oxymethyl ester, in the form of a faint yellow foam, which is processed without further purification.

Rf = 0.33 /N-acetyl-4-0,6-o-isopropylidene-muramyl-L-alanyl-D-glutamine pivaloyloxymethyl ester/ (methylene chloride: methanol: water = 70:30:5).

Step 19, 4:

A solution of 5.5 g (approx. 8.5 mmol) of crude N-acetyl-4-0,6-o-isopropylidene-muramy1-L-alanyl-D-glutamine-pivaloyloxy-methyl ester in 100 ml of glacial acetic acid-water ( 3:2), stirred for 4.5 hours at room temperature. The slightly yellow solution is then evaporated in high vacuum at 40°C, and the residue obtained is purified by column chromatography on 500 g silica gel ("type 60", pure, Merck; 0.063-0.2 mm) in the system methylene chloride:methanol:water (70:30:5) (10 ml fractions). Fractions 52-65 are collected and evaporated in high vacuum at 30°C. N-acetyl-muramyl-L-alanyl-D-glutamine pivaloyloxymethyl ester is obtained, which dissolves

in 60 ml double distilled water. The solution is filtered through a millipore filter ("Nalgen S"; 0.2 µm) and then lyophilized under high vacuum. N-acetyl-muramyl-L-alany1-D-glutamine-pivaloyloxymethyl ester is obtained as a colorless powder which still contains 1.21 mol of water.

C25H42<N>4°13* 1.21 H2° (628'5°)

Calculated C 47.78 H 7.16 N 8.91 0 36.18 H20 3.48

Found C 48.09 H 7.15 N 9.02 0 36.61 H20 3.48

/a/20** ^39.5 + 0.1° (c=0.443; water),

Rf = 0.61o (methylene chloride:methanol:water = 70:30:5).

Example 20:

By the methods described in the present description

the following connections are obtained:

a) N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-N-methyl-alanyl-D-isoglutamine-benzhydryl ester, b) N-acetyl-1,4,6-tri-O-acetyl- muramyl-α-amino-isobutyryl-D-isoglutamine-benzhydryl ester and c) N-acetyl-1,4,6-tri-O-acety1-desmethylmuramyl-L-alanyl-D~y-methoxycarbonyl-isoglutamine-benzhydryl ester.

Example 21:

Female MF-2f SPF mice with a body weight of 14-16 g are infected intranasally under light anesthesia with a mixture of equal parts of diethyl ether, ethanol and chloroform at lethal doses (approximately one LDg0_<g>Q<;><1->4 "plaque forming units" (PFU)) in the form of 0.05 ml of a suspension of influenza A/Texas/1/77 (virus A) or of influenza B/Hong Kong 15-72 (virus B) viruses ( mouse-adapted strains).

Each mouse from groups of a total of 10 of these mice is supplied

at the time indicated below (days) starting on the day of infection 1 time (single dose) of the amount of the relevant active substance indicated in Table 1 in 0.05 or 0.2 ml of a 0.005% by weight solution of carboxy- methylcellulose sodium salt in doubly distilled, pyrogen-free water in the case of intranasal or oral administration of the type specified in Table 1. 20 of the above-mentioned infected mice serve as controls, i.e. they received a placebo (0.005% by weight solution of carboxymethylcellulose sodium salt).

The intranasal supply 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-desmethyl-muramyl-L-alanyl-D-isoglutamine-benzhydryl ester.

Example 22:

Non-aqueous single dose for nasal administration: Composition: 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-

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 common single-dose nasal applicator, to which a propellant dose is attached before use.

Example 23:

Nasal drops:

Composition:

1,4,6-tri-O-acetyl-N-propionyl-desmethyl-muramyl-L-alanyl-D-isoblutamine-benz-

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 part thereof or multiple thereof is filtered through a membrane filter and filled into a cleaned container. Suitable containers are e.g.

a) glass or plastic containers (to 5 ml or 10 ml), which have a glass or plastic pipette with a

elastomer pipette suction,

b) collapsible bottles made of plastic with a riser and a spray head made of plastic, c) single-dose containers made of plastic (content 2-3 drops) or d) glass or plastic bottles equipped with a standardized pump dosing spray made of plastic (without propellant gas).

Example 2 4:

Nasal ointment.

Composition:

1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-iso-

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

Example 25:

Production of 1000 tablets, containing 0.5% active substance.

Composition per 1000 tablets:

1,4,6-tri-O-acetyl-N-propionyl-des-methylmuramyl-L-alanyl-D-isoglutamine-

The active substance and 15 g of lactose are mixed together. The obtained premix is mixed with 28 g of lactose and 47 g of corn starch. With the obtained mixture and an aqueous solution of "Pharmacoat" a granulation-ready mass is prepared which is granulated, dried and ground. To this, 5 g of corn starch, aerosil and magnesium stearate are mixed and the mixture is 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 26:

The methods mentioned in the present description yield the following compounds: N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-alanyl-D-isogluta-minyl-L-a-(/2-benzoyloxy-carbonylamino- ethyl ()-sulfony1-methyl)-glycine benzyl ester, N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-alanyl-D-isoglutami-nyl-L-O-acetyl-serine benzyl ester,

1,4,6-tri-O-acetyl-N-benzoyl-desmethylmuramy1-L-alanyl-D-glutamic acid dicholinester,

N-propionyl-1,4,6-tri-O-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester, 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl- D-glutamic acid α-n-butyl ester γ-benzyl ester, 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid dibenzyl ester, N-acetyl-1,4,6- tri-O-propionyl-muramyl-L-α-aminobutyryl-D-isoglutamine-benzhydryl ester,

1,4,6-tri-O-acetyl-N-benzoyl-desmethylmuramyl-L-alanyl-D-isoglutamine benzyl ester and

1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid dipivaloyloxymethyl ester.

Example 27:

Analogous to example 1, you get from 250 mg (approx. 0.098 mmol)

crude 4,6-di-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid dipivaloyloxymethyl ester (a,B mixture)

and 2.0 mL (21.0 mmol) acetic anhydride in 10 mL absolute pyridine (18 hours, room temperature) 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alany1-D-glutamic acid dipival -oyloxy methyl ester (a , B mixture);

Rf = 0.43 (chloroform:ethanol = 95:5),

Rf = 0.56 (chloroform:methanol = 9al),

Rf = 0.72 (chloroform:methanol = 4:1).

The starting material is obtained in the following way:

Step 27. 1:

To a solution of 9.78 g (34.1 mmol) of N-tert-butyloxy-carbonyl-L-alanine-N-hydroxy-succinimide ester in 200 ml of absolute N,N-dimethylformamide is added at room temperature with stirring 15.9 g (34.1 mmol) D-glutamic acid-(Ca)-pivaloyloxymethyl ester-()-benzyl ester tosylate and 3.75

ml (34.1 mmol) N-methylmorpholine. After 18 hours of heating at room temperature, it is evaporated in vacuum at 40°C. The crude product obtained (yellow oil) is purified by column chromatography on 800 g silica gel ("type 60", pure, Merck; 0.063-0.2 mm)

in the system methylene chloride-acetic acid ethyl ester (85:15) (10

ml fractions). Fractions 126-230 are collected and evaporated in vacuum at 30°C. This gives N-tert.-butyloxycarbonyl-L-

alanyl-D-glutamic acid-(C^)-pivaloyloxymethyl ester-(C )-benzyl ester as yellowish oil:

/a/p = -10.6 + 0.1° (c = 1.35; methylene chloride),

Rf = 0.29 (methylene chloride; ethyl acetate = 85:15),

Rf = 0.73 (methylene chloride:methanol = 9:1),

Rf = 0.78 (methylene chloride:methanol = 5:1).

Step 27, 2:

To a solution of 4.0 g (7.65 mmol) of N-tert-butyloxy-carbonyl-L-alanyl-D-glutamic acid-(C )-pivaloyloxymethyl-ester-(C )-benzyl ester in 50 ml of absolute acetic acid ethyl ester man at 0°C 100 ml approx. 5N hydrochloric acid in acetic acid ethyl ester and stir for 1 hour at 0°C. It is then evaporated in a high vacuum at 30°C and the residue obtained is taken up several times in acetic acid ethyl ester and evaporated again.

A colorless foam is obtained which is dissolved in 50 ml of acetic acid ethyl ester. After adding 250 ml of pentane and cooling to -10°C, L-alanyl-D-glutamic acid-(Ca)-pivaloyl-oxymethyl ester-(C)-benzyl ester hydrochloride is obtained as colorless crystals with m.p. 73-74°C, which still contains 0.63 moles of water;

/a/p = +23.5+0.1° (c=1.150; methanol),

Rf = 0.20 (methylene chloride-methanol =9:1),

Rf = 0.49 (methylene chloride-methanol = 5:1),

Rf = 0.87 (methylene chloride: methanol: water = 70:30:5).

Step 27, 3:

To a suspension of 2.99 g (5.0 mmol, still containing sodium chloride; 1.673 mmol/g) of 1-a-O-benzy1-4,6-0,0-isopropylidene-N-propionyl-desmethylmuramic acid sodium salt in 50 ml absolute N,N-dimethylformamide, 1.34 g (6.5 mmol) N,N-dicyclohexylcarbodiimide, 1.03 g (6.5 mmol) 1-hydroxytriazole and 2.35 g ( 5.0 mmol) L-alanyl-D-glutamic acid-(C )-pivaloyloxymethyl ester-(C )-benzyl ester hydrochloride and the mixture is then stirred for 22 hours at room temperature. The suspension is then filtered (N,N-dicyclohexylurea) and the filtrate is evaporated in high vacuum at 40°C. The residue, a yellowish foam, is taken up in 100 ml of ethyl acetate and the solution obtained is washed successively with 50 ml of 2N citric acid, 10% sodium bicarbonate solution and water. The acetate fractions are collected, dried over sodium sulfate and evaporated in vacuo. The crude product obtained is purified by column chromatography on 400 g silica gel ("type 60", pure, Merck; 0.063-0.2 mm) first with chloroform (course 2 liters), then in the system chloroform-ethanol (95:5) (10 ml fractions). Fractions 93-122 are collected and evaporated in vacuo. Weakly contaminated l-α-0-benzyl-4,6-0,0-isopropylidene-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid-(C ot )-pivaloyloxymethyl ester-(C )-benzyl ester is still obtained as a colorless foam which is processed without further purification;

Rf = 0.55 (chloroform:ethanol = 9:1),

Rf = 0.63 (chloroform:methanol = 9:1).

Step 27, 4:

To a solution of 830 mg (1.0 mmol) 1-α-O-benzyl-4,6-0,O-isopropylidene-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid-(Cq)-pivaloyloxymethyl ester-(C y)-benzyl ester

in 28.5 ml of absolute methylene chloride is added at 0°C

1.5 ml of trifluoroacetic acid and stir the mixture for 1.5 hours at 0°C. The colorless solution obtained is then evaporated in high vacuum at 40°C to dryness. The residue is crystallized from chloroform-methanol-diethyl ether (5:1:70). 1-a-O-benzyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid-(Cq)-pivaloyloxymethylester-(C )-benzyl ester is obtained as colorless crystals; m.p. 148-149°C;

Rf = 0.33 (chloroform:methanol =9:1),

Rf = 0.67 (chloroform:methanol =4:1),

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

Step 27, 5:

400 mg (0.5 mmol) of 1-α-O-benzyl-N-propionyl-desmethylmuramyl-L-alany1-D-glutamic acid-(Ca)-pivaloyloxymethyl ester-(C y)-benzyl ester are hydrated in 80 ml of tetrahydrofuran-methanol (4: 1) with 200 mg of 10% palladium charcoal as catalyst at room temperature and under normal pressure. After complete hydration (1 hour), the catalyst is filtered off and the filtrate is evaporated under high vacuum. The residue, a colorless foam, is dissolved in a mixture of 30 ml of tetrahydrofuran and 1 ml of water, and the resulting solution is mixed at room temperature with a solution of 96 mg (0.25 mmol) of cesium carbonate. After standing for 1/2 hour at room temperature, the colorless solution is evaporated in vacuo. The residue is then mixed twice with 100 ml of methanol and once with 100 ml of N,N-dimethylformamide and evaporated again. Slightly contaminated 1-α-O-benzyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid-(C )-pivaloyloxymethyl ester cesium salt is still obtained as a colorless foam, which is worked up without further purification;

Rf = 0.15 (chloroform:methanol =4:1),

Rf = 0.35 (chloroform:methanol = 7:3),

Rf = 0.45 (chloroform:methanol:water = 70:30:5).

Step 27, 6:

To a solution of 420 mg (0.5 mmol) of 1-α-O-benzyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid-(C )-pivaloyloxymethyl ester cesium salt in 15 ml of N,N-dimethylformamide is added at room temperature with stirring 150

mg (145 (1.0 mmol) pivalic acid chloromethyl ester and 150

mg (1.0 mmol) sodium iodide. The resulting mixture is stirred for 18 hours at room temperature, then 145 µl (1.0 mmol) of pivalic acid chloromethyl ester and 150 mg (1.0 mmol) of sodium iodide are added again and stirred for a further 48 hours. The mixture is then evaporated under high vacuum at 30°C The crude product is purified by column chromatography twice on 200 and 250 silica gel ("type 60", pure, Merck; 0.062-0.2 mm) in the system chloroform-methanol (9:1) and chloroform:methanol (95:5). The fractions containing the desired product is collected and evaporated in vacuo to obtain 1-a-O-benzyl-N-propionyl-desmethylmuramyl-L-alanyl-D-g.lutamic acid dipivaloyloxymethyl ester;

Rf = 0.25 (chloroform:methanol = 9:1),

Rf = 0.35 (chloroform:ephanolI= 9:1),

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

Step 27, 7:

Analogously to example 1, one obtains from 80 mg (0.098 mmol) 1-α-O-benzyl 1-N-propionyl-desmethylmuranyl-L-alanyl-D-glutamic acid dipivaloyloxymethyl ester and 0.8 ml (8.4 mmol) acetic anhydride in 8 ml absolute pyridine (18 hours at room temperature) 4,6-di-O-acetyl-1-a-O-benzyl-N-peopionyl-desmethylmura-myl-L-alanyl-D-glutamic acid re-dipivaloyloxymethyl ester;

Rf = 0.50 (chloroform:ethanol = 95:5),

Rf = 0.65 (chloroform:ethanol = 9:1),

Rf = 0.77 (chloroform:methanol = 9:1).

Step 27, 8:

140 mg (0.098 mmol) of 4,6-di-O-acetyl-1-α-O-benzyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid dipivaloyloxymethyl ester is hydrated in 80 ml of dimethoxyethane-water (9:1) with palladium charcoal ( Degussa, "E 101 N") as a catalyst for 20 hours at room temperature and under normal pressure. The catalyst is then filtered off and the filtrate is evaporated in high vacuum at 30°C. One obtains crude 4,6-di-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid dipivaloyloxymethyl ester (a,B mixture), which is processed without further purification;

R 2 = 0.38 (chloroform:methanol = 19 µl),

Rf = 0.64 (chloroform:methanol:water = 70:30:5),

Rf = 0.68 (chloroform:methanol = 4:1).

Example 28:

Analogous to example 1, one obtains from 174.0 mg (0.25 mmol) N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine benzoyloxymethyl ester, which still contains traces of N-methyl-morpholine hydrochloride, in 1.7 ml absolute pyridine with 0.1 ml (1.04 mmol) acetic anhydride while adding

of a little 4-dimethylamino-pyridine 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine-

benzoyloxymethyl ester (a , B mixture) as colorless crystals;

Temp. 177-178°C (from methylene chloride:diethyl ether = 1:1),

Rf = 0.45 (chloroform:methanol = 4:1),

Rf = 0.64 (chloroform:methanol ) 5:3).

The starting material is obtained in the following way:

Step 28. 1:

A solution of 18.9 g (0.1 mol) of N-tert-butyloxycarbonyl-L-alanine in 50 ml of N,N-dimethylacetamide is mixed with 14.9 ml (0.1 mol) of 1,8-diazabicyclo/5.4 .0/undec-7-ene and 36.2 ml (0.45 mol) of diiodomethane and stirred for 7 hours at 22°C. The solution is evaporated under high vacuum and a suspension of 36.6 g (0.3 mol) benzoic acid, 6.1 g (0.05 mol) 4-dimethylaminopyridine and 37.3 ml 1,8-diazabicyclo-1_ 5 is added to the residue. . 4. 0/undec-7-ene in 75 ml of N,N-dimethylacetamide. The clear solution that occurs after 40 minutes is allowed to stand for 16 hours at 22°C. The mixture is decanted with a mixture of -0.3 molar dipotassium hydrogen phosphate solution and acetic acid ethyl ester, the organic layer is separated, washed with further phosphate solution, dried over sodium sulphate and evaporated. Digestion of the residue with pentane, then pentane-ethyl ether (3:1) gives N-tert-butyloxycarbonyl-L-alanine-benzoyloxymethyl ester;

R^ = 0.68 (chloroform: ethyl acetate = 9:1).

Step 28, 2:

3.72 g (11.5 mmol) of N-tert.-butyloxycarbonyl-L-alanine-benzyloxy-methyl ester are dissolved to a clear solution at 0°C in 100 ml of a 5 molar hydrogen chloride solution in acetic acid ethyl ester and allowed to stand for 2 hours at 0°C. At 0°C, the solution is added to 200 ml of diethyl ether-pentane (1:1)

and after 20 minutes of stirring at 0°C, the precipitated gel-like crude product is filtered off. It is stirred for 2 hours at 22°C in 350 ml acetic acid ethyl ester-diethyl ether-pentane (1:1:1), and then filtered off. L-alanine benzoyloxymethyl ester hydrochloride is obtained; m.p. 99-100°C.

Step 28, 3:

To a solution of 506 mg (1.0 mmol) of N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine, containing 0.77

mol of water, in 10 ml of absolute N,N-dimethylformamide, 268 mg (1.3 mmol) of N,N-dicyclohexylcarbodiimide and 207 mg (1.3 mmol) of 1-hydroxy-benzotriazole are added at room temperature with stirring. After 2 hours, the obtained colorless suspension is mixed step by step with 269 mg (1.0 mmol) of L-alanine benzoyloxymethyl ester hydrochloride and 110 (1.0 mmol) of N-methylmorpholine and is then stirred further at room temperature. After 2 hours, the precipitated N,N-dicyclohexylurea is filtered off and the filtrate is evaporated in high vacuum at 30°C.

The obtained slightly yellow residue (foam) is suspended

in 25 ml ethyl acetate and this suspension is stirred for 1 hour at room temperature. The crude product obtained in the form of colorless crystals is further purified by column chromatography on 150 g silica gel ("type 60", pure, Merck; 0.063-0.2 mm) in the system chloroform-methanol-water (70:30:5)

(5 ml fractions). Fractions 25-35 are collected and evaporated under high vacuum. Almost pure N-propionyl-desmethyl-muramyl-L-alanyl-D-isoglutaminyl-L-alanine benzoyloxyrnetyl ester (a,B mixture) is obtained as a colorless foam, which still contains traces of N-methyl-morpholine hydrochloride and which are processed without further purification;

Rf =0.36 (chloroform:methanol = 7:3),

Rf = 0.41 (chloroform:methanol:water = 70:30:5),

Rf = 0.74 (acetonitrile:water = 3:1).

Example 29:

1.95 g (2.18 mmol) of N-acetyl-muramyl-L-alanyl-D-isoglutaminyl-L-(C<a>)-(/2-benzyloxycarbonylamino-ethyl7_sulfonylmethyl)-glycine-benzyl ester are dissolved in 40 ml abs . pyridine and is peracetylated with 30 times the molar amount of acetic anhydride (30 hours, RT). The gel-like mass is brought into solution with a little water, the whole is evaporated on a rotary evaporator in high vacuum at 30°C to a small volume, and lyophilized after adding abs. dioxane. The crude product is subjected to flame chromatography (145:1; 25 ml fractions; 0.5 bar on silica gel (grain size 0.04-0.063 mm) first with chloroform, then with chloroform-methanol mixtures (98:2 to 1:1). The fractions which contains the product collects, dissolves in

90% dioxane (80 ml) and lyophilized after sterile filtration (0.2 µm; PTFE). N-acetyl-1, 4, 6-tri-O-acety 1-muramyl-L-alanyl-D-isoglutaminyl-L-(C<a>)-(/2-benzyloxycarbonylamino-ethyl/-sulfonyl-methyl) is obtained )-glycine benzyl ester (a , B - mixture) as slightly yellowish powder, containing 1.27 moles of water;

/a/D = +36.6 +1° (c = 1.030; dimethylformamide),

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

Rf = 0.53 (n-butanol:acetic acid:water = 70:7.5:21),

Rf = 0.61 (acetonitrile:water = 3:1).

The starting material is obtained in the following way:

Step 29, 1:

21.5 g (100 mmol) of a 31.4% hydrogen peroxide solution. After 30 hours of stirring, 1.5 g of palladium charcoal (5%) are introduced to decompose excess peroxide, whereby a strong evolution of gas occurs. After stirring for 3 hours, the catalyst is removed and the reaction solution is evaporated to dryness on a rotary evaporator at 30°C. The oily residue is dissolved in 11 ml of acetone and brought to crystallization by adding 20 ml of diethyl ether. After recrystallization from the same mixture, Ne<->benzyloxycarbonyl-L-thialysine-S,S-dioxide-benzyl ester hydrochloride is obtained in the form of colorless needles; m.p. 141-142° (dec.); m.p. for the p-toluene sulphonate 130-134°C (dec.);

Rf = 0.50 (chloroform:methanol:water = 70:30:5),

Rf:= 0.89 (methyl ethyl ketone:pyridine:water = 65:5:20),

Rf 0.71 (n-butanol:acetic-water = 3:1:1).

Step 29, 2:

2.95 g (6 mmol) of N-acetyl-muramyl-L-alanyl-D-isoglutamine and 2.74 g (6 mmol) of Ne<->benzyloxycarbonyl-L-thialysine-S,S-dioxide benzyl ester hydrochloride are connected in ordinary manner (see step 30.1) together with each other by the dicyclohexylcarbodiimide-1-hydroxy-benztriazole method. After a total of 51 hours of stirring at RT, the reddish suspension is mixed with 200 ml of acetic acid ethyl ester, the insoluble components are filtered off after 1 hour of stirring and the filtrate is evaporated to dryness on a rotary evaporator in high vacuum at 30°C. The residue is mixed with 100 ml of acetic acid ester and water and stirred for 1 hour. The gel-like material is filtered off, dried and then distributed between 80 ml of the upper and lower phases of mutually saturated n-butanol-water (1:1). After 1 hour of stirring, the insoluble material is sucked off, washed and dried. N-acetyl-muramyl-L-alanyl-D-isoglutaminyl-L-(C<a>)-(/2-benzyloxycarbonylamino-ethyl/-sulfonylmethyl)-glycine-benzyl ester (a , 8-mixture) is obtained in the form of a white powder containing 1.63 moles of water;

Rf = 0.60 (chloroform:methanol:water = 70:30:5),

Rf = 0.83 (acetonitrile:water = 3:1).

Example 30:

1.50 g (2.24 mmol) of N-acetyl-muramyl-L-alanyl-D-isoglutaminyl-L-serine benzyl ester are suspended in 40 ml of abs. pyridine and is peractylated with 40 times the molar amount of acetic anhydride. The gel-like material obtained after 24 hours of stirring at RT is dissolved by the addition of 5 ml of water, the whole is evaporated on a rotary evaporator in high vacuum (30°C) to approx. 5 ml, mixed with double-distilled water and lyophilized. The raw product is absorbed in 50

ml of warm acetic acid ethyl ester and is precipitated by the addition of 5 parts by volume of diethyl ether (2 times). The precipitate is dissolved in chloroform-methanol (1:1), sterile-filtered (0.2 (im, PTFE),

is recorded after evaporation of the solvent in abs. dioxane and freeze-dried. N-acetyl-1,4,6-O-triacetyl-muramyl-L-alanyl-D-isoglutaminyl-L-O-acetyl-serine-benzyl ester hydrate is obtained

(a,6 mixture) as colorless powder;

--2 0

/a/D= +26.5 + 0.9° (c = 1.096; chloroform:methanol = 1:1),

Rf = 0.10 (chloroform:isopropanol = 7:2),

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

Rf = 0.33 (n-butanol:acetic acid:water = 75:7.5=21).

The starting material is obtained in the following way:

Step 30, 1:

5.00 g (10.15 mmol) N-acetyl-muramyl-L-alanyl-D-isoglutamine, 2.00 g (13.2 mmol) 1-hydroxybenztriazole (containing 12% water) and 2.35 g (10 .15 mmol) L-serine benzyl ester hydrochloride is dissolved in 50 ml of dimethylformamide. Then 1.17 ml (10.66 mmol) of N-methylmorpholine and finally 2.72 g (13.2 mmol) of N,N'-dicyclohexylcarbodiimide are added. After stirring for 24 hours at room temperature, the reddish suspension is mixed with 100 ml ethyl acetate and stirred for 1 hour. The insoluble dicyclohexylurea is filtered off and the filtrate is evaporated in high vacuum on a rotary evaporator at 30°C. The yellow oil obtained is purified on silica gel (60:1; 15 ml fractions) in the system chloroform:methanol:water = 70:30:5. The fractions containing the product are collected and lyophilized after evaporation of the solvent from tert.-butanol-water (9:1). N-acetyl-muramyl-L-alanyl-D-isoglutaminyl-L-serine benzyl ester (α,6 mixture) is obtained as a colorless powder containing 1.87 mol of water;

--70

/a/546nm= 12.4 +1° (c = 0.978; water),

R^ = 0.59 (ethyl acetate: n-butanol: pyridine-acetic acid: water = 42:21:21:6:10).

R^ = 0.41 (chloroform:methanol:water = 70:30:5).

Example 31.

0.36 g (0.535 mmol) of N-acetyl-muramy-1-L-N-methylalany-1-D-isoglutamine-benzhydryl ester (a,B mixture) is dissolved in 4

ml abs. pyridine and after the addition of 0.15 ml (16 mmol) of acetic anhydride is allowed to stand for 3 hours at room temperature. The yellowish solution is evaporated under high vacuum at 30°C, taken up in 10

ml of water and lyophilized. The crude product is purified by chromatography

on silica gel (1:180) in the system chloroform-isopropanol (9:1)

(0.8 ml fractions). The uniform fractions are pooled and lyophilized from dioxane. N-acetyl-1,4,6-tri-O-acety1-muramyl-L-N-methyl-alanyl-D-isoglutamine-benzhydryl ester (a,B mixture) is obtained as a colorless powder;

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

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

The starting material is obtained in the following way:

Step 31, 1:

0.50 g (0.99 mmol) of N-acetyl-muramyl-L-N-methyl-alanyl-D-isoglutamine (a,B mixture), dissolved in methanol, is esterified with an excess of diphenyldiazomethane. After standing for 2 hours, evaporate to dryness and the residue is triturated several times with petroleum ether. It is suctioned off, the residue is taken up in a little methanol and the material is precipitated with tenfold the amount of diethyl ether:petroleum ether (1:1). N-acetyl-muramyl-L-N-methylalanyl-D-isoglutamine-benzhydryl ester is obtained as a colorless powder;

Rf = 0.76 (acetonitrile:water = 3:1),

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

Example 32:

0.50 g (0.99 mmol) of N-acetyl-muramyl-a-aminoisobutyryl-D-isoglutamine (a,B mixture) is esterified analogously to step 31.1

with diphenyldiazomethane. The crude product /Rf = 0.48 (chloroform: methanol:water = 70: 30: 5]_/ is peracetylated analogously to example 31. The reaction solution is evaporated in vacuo to dryness, taken up in water and lyophilized. Purification takes place on silica gel (1:100) in chloroform-methanol (9:1; 1 ml fractions).

The purified fractions are pooled, 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-O-acetyl-muramyl-a-aminoisobutyryl-D-isoglutamine-benzhydryl ester (a,B mixture) is obtained as a colorless powder, containing 0.73 mol of water;

--7 0

/^/q +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 33:

33 mg of N-acetyl-desmethylmuramyl-L-alanyl-D-y-methoxycarbonyl-isoglutamine-benzhydryl ester (a,B mixture) is peracetylated analogously to example 31. After lyophilization from abs. dioxane, N-acetyl-1,4,6-tri-O-acetyl-desmethylmura-myl-L-alanyl-D-y-methoxycarbonyl-isoglutamine-benzhydryl ester (a,B mixture) is obtained as a colorless powder:

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

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

The starting material is obtained in the following way:

Step 33, 1:

0.216 g (0.40 mmol) of N-acetyl-desmethylmuramyl-L-alanyl-D-Y-carboxy-isoglutamine (a,B mixture) is esterified analogously to step 31.1 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 and decomposition products originating from diphenyldiazomethane is separated by chromatography on silica gel (1:100) in the system chloroform-methanol-water (70:30:5). After two rounds of chromatography, N-acetyl-desmethylmuramyl-L-alanyl-D-y-benzhydryloxycarbonyl-isoglutamine-benzhydryl ester (a,8 mixture) is obtained

as color resin with the R^ 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

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

Step 3 3. 3: N-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester esterified in methanolic solution as usual with a solution of diazomethane in diethyl ether. After evaporation, N-acetyl-desmethylmuramyl-L-alanyl-D-y-methoxycarbonyl-isoglutamine-benzhydryl ester (a,B mixture) is obtained as a colorless resin;

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

Example 34:

1.70 g (02.53 mmoL) N-acetyl-muramyl-L-a-aminobutyryl-D-isoglutamine-benzhydryl ester (a,B mixture), dissolved in 25 ml abs. pyridine, is perpropionylated analogously to example 31 (90 equivalents of anhydride). After standing for 3 hours at RT, 30 ml of water is added and the whole is evaporated in HV at 30°C to dryness. The resinous residue is taken up in 80 ml abs. dioxane, filtered PTFE; 0.2 µm) and lyophilized. N-acetyl-1,4,6-tri-O-propionyl-muramyl-L-a-amino-butyryl-D-isoglutamine-benzhydryl ester (a,B mixture) is obtained as a colorless powder, which contains 0.79 mol of water;

/a/p = +52.5 + 3.7° (c = 0.270; methanol),

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

Rf = 0+32 (chloroform:isopropanol:acetic acid = 30:8:2),

Rf = 0.87 (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:

2.03 g (4 mmol) of N-acety1-muramy1-L-α-aminobutyryl1-D-isoglutamine (a,B mixture) is transferred to the benzhydryl ester analogously to step 31.1. After normal work-up, N-acetyl-muramyl-L-a-aminobutyryl-D-isoglutamine-benzhydryl ester (a,B mixture) is obtained as a colorless powder:

/a/p = +41.8 + 1.8° (c = 0.548; methanol),

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

R^ = 0.78 (acetonitrile:water = 3:1).

Example 35.

Analogously to example 1, one obtains from 2.0 g (2.94 mmol) N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester (a,B mixture), which contains 1.17 mol of water, in 20 ml abs. pyridine with 1.68 mL (13.06 mmol) propionic anhydride (puris-simum; d = 1.012) 1,4,6-tri-O-propionyl-N-propionyl-desmethyl-muramyl-L-alanyl-D-isoglutamine- benzhydryl ester (a,B mixture) as a colorless lyophilisate, which still contains 0.68

moles of water; m.p. 156-157°, -

--20

/a/p = + 39.02 + 2.1° (c = 0.467; methanol),

Rf = 0.40 (chloroform:ethanol = 9:1),

R^= 0.48 (chloroform:methanol = 9:1),

Rf = 0.88 (chloroform:methanol = 4:1).

Example 36:

Analogously to example 1, one obtains from 2.0 g (3.1 mmol) N-propionyl-1-desmethylmuramyl-L-alanyl-D-glutamic acid-(C )-n-butyl ester-(C )-benzyl ester (mainly a -anomer, which still contains 0.41 mol of water, in 20 ml of abs. pyridine with 1.14

ml (12.0 mmol) acetic anhydride 1,4,6-tri-O-acetyl-N-propiony1-desmethylmuramyl-L-alanyl-D-glutamic acid-(C )-n-butyl-a

ester-(C )-benzyl ester as a colorless lyophilisate which

1

divided contains 0.73 mol of water. According to the H-NMR spectrum (360 MHz), the compound exists almost exclusively in the form of the α-anomer; "mp". 62-64;

/a/p = +36.4 + 3.6° (c = 0.280; methanol),

R^ = 0.43 (chloroform:methanol = 9:1),

R^= 0.55 (chloroform:ethanol = 9:1),

Rf = 0.85 (chloroform:methanol = 4:1).

Example 37:

Analogous to example 1, one obtains from 0.57 g (0.83 mmol) N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid dibenzyl ester (a,B mixture) containing 0.89 mol of water, in 8 ml of abs . pyridine with 0.36 ml (3.807 mmol) acetic anhydride 1,4,6-tri-O-acetyl-N-propiony1-desmethylmuramyl-L-alany1-D-glutamic acid dibenzyl ester (a,B mixture) in the form of colorless crystals ; m.p. 141-142° (from chloroform:diethyl ether = 1:10), /a/p°= +22.7 + 2.2° (c = 0.454; methylene chloride),

Rf = 0.42 (chloroform:methanol = 9:1),

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

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

The starting material is produced in the following way:

Step 37, 1:

0.74 g (1.95 mmol) of 4,6-0,0-isopropylidene-N-propionyl-desmethylmuramic acid sodium salt is suspended in 20 ml of N,N-dimethylformamide, then 0.342 g (2.145 mmol) of 1-

hydroxybenzotriazole, 0.442 g (2.145 mmol) N,N-dicyclohexylcarbodiimide and 0.85 (1.95 mmol) L-alanyl-D-glutamic acid dibenzyl ester hydrochloride and the mixture is stirred overnight at room temperature. The precipitated N,N-dicyclohexylurea is then filtered off and the filtrate obtained is evaporated in high vacuum to dryness at 30°C. The residue is taken up in 150 ml of ethyl acetate and washed successively with 50 ml of saturated sodium bicarbonate solution,

2N citric acid, saturated sodium bicarbonate solution and water. The organic phases are collected, dried over sodium sulfate and evaporated under high vacuum at 30°C. 4,6-0,O-isopropylidene-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid dibenzyl ester (a,B mixture) is obtained, which contains some N-propionyl-desmethylmuramyl-L-alanyl-D- glutamic acid dibenzyl ester (a,B mixture) as well as N,N-dicyclohexylurea, and is further processed without further purification.

Step 37, 2:

3.3 g of crude 4,6-0,O-isopropylidene-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid dibenzyles (a,B mixture) gives

stand overnight in 30 ml 50% acetic acid. The resulting solution is then filtered and the filtrate is evaporated in high vacuum at 30°C. The residue is purified by column chromatography on

150 g silica gel ("type 60", pure, Merck; 0.063-0.2 mm)

in the system chloroform:ethanol (9:1; 10 ml fractions). Fractions 56-95 are collected and evaporated under high vacuum. The residue is then dissolved in 80 ml of methanol and the slightly cloudy solution thus obtained is filtered through a millipore filter ("Fluoropore", PTFE, 0.2 µm). The clear filtrate is evaporated in vacuo at 40°C. The residue is then dissolved in 10 ml abs. methanol previously filtered through a millipore filter ("Fluoropore", PTFE, 0.2 (im), and cry-

tally by adding 100 ml of abs. diethyl ether as well

is filtered through a millipore filter, and washed with filtered, abs. diethyl ether. N-propionyl-desmethylmuramy1-L-alany1-D-glutamic acid dibenzyl ester (a,B mixture) is obtained in the form of colorless crystals containing 0.89 mol of water; m.p. 146-147°,

--2 0

/a/p = 14.6+2.1° (c = 0.478; methanol),

Rf = 0.20 (chloroform:methanol = 9:1),

Rf = 0.65 (chloroform:methanol = 4:1),

Rf = 0.81 (chloroform:methanol 0 7:3).

Example 38:

Analogous example 1 is obtained with pyridine and acetic anhydride

from N-benzoy1-desmethylmuramy1-L-alanyl-D-glutamic acid--dicholinester dichloride, 1,4,6-tri-O-acetyl-N-benzoy1-desme-ty1-muramy1-L-alany1-D-glutamic acid- dicholinester dichloride. The starting material is obtained in the following way:

Step 38, 1:

As described in Synthesis 1982, 138, one obtains from D-glutamic acid dimethyl ester hydrochloride and excess choline chloride with titanium tetraethyl ester as catalyst in acetonitrile at 80°C, D-glutamic acid dicholine ester dichloride hydrochloride.

Step 38, 2:

Analogously to step 6.3, from N-benzoyl-desmethyl-muramyl-L-alanine and D-glutamic acid dicholinester dichloride hydrochloride with dicyclohexylcarbodiimide N-benzoyl-desmethylmuramyl-L-alany1-D-glutamic acid dicholinester dichloride is obtained.

Example 3 9:

Analogous to example 1, one obtains from N-benzoyl-desmethyl-muramyl-L-alanyl-D-isoglutamine-benzhydryl ester with n-hexanoyl chloride, pyridine and 4-dimethylamino-pyridine N-benzoyl-1,4,6-tri-O-n -hexoyl-L-alanyl-D-isoglutamine benzhydryl ester.

Claims (10)

1. Process for the preparation of compounds of formula I, wherein the hexose moiety is derived from D-glucose, D-mannose or D-galactose, n represents 0 or 1, and r\ R<4> and R^ independently represent 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> stands for hydrogen, R g for hydrogen or unsubstituted or with phenyl, hydroxy, mercapto or lower alkylthio substituted lower alkyl, R 9 and R 12 stand independently of each other for hydroxy, amino, C^ _^Q - alkoxy, aryl lavereal oxy, alkanoyloxylavereal oxy with up to 16 C atoms, aroyloxylavereal oxy, 3-cholesteryloxy or 2-trimethylammonioethyloxy, R" ^ represents hydrogen, carboxy, lower alkoxycarbonyl or aryllower oxycarbonyl, and R"'""'" represents hydrogen or unsubstituted or with amino, hydroxy, lower alkanoylamino, lower alkanoyloxy, 2-benzyloxycarbonylamino- ethylsulfinyl, 2-lower-alkoxycarbonylamino-ethylsulfinyl, 2-lower-alkoxycarbonylamino-ethylsulfinyl, 2-lower-alkoxycarbonylamino-ethylsulfonyl or guanidino-substituted lower alkyl, under the condition that at least one of the residues one R and R is for different from hydroxy, amino and C^ _^ - alkoxy or R"^ is different from hydrogen, carboxy and alkoxycarbonyl with up to 7 C atoms in the alkoxy part, and salts of such compounds with at least one salt-forming group, characterized in that mana) reacts a compound of formula II v, x. t,12 „2a „4a „6a . « c in which at least one of the residues R , R , R and R stands for hydrogen and the rest of these residues have the meaning of r\ the group R2-C(=0)-, R4 respectively R^, and the rest of the substituents have the above given meanings, whereby in a compound of formula II present free functional groups, with the exception of groups which are to participate in the reaction, can, if necessary, be protected with easily cleavable protective groups, with an acylating agent which transfers the residue r\ R2-C(=0) -, R4 or R^ to be introduced, and then, if necessary, cleave off the protective groups present, orb) react a compound of formula III, in which the substituents have the meanings indicated above, or a reactive derivative thereof, with a compound of formula IV, where X stands for a reactive, esterified hydroxy group, and the other substituents have the above-mentioned meanings, whereby in a compound of formula IV present free-functional groups, with the exception of X, can, if necessary, be protected with easily removable protective groups, and then if necessary cleaves off protecting groups present, orc) reacts a compound of formula V in which q, r, s and t independently stand for 0 or 1 and where 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 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 means of 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 with a reactive derivative thereof, and if necessary cleaves off any protective groups present, ord) for the preparation of a compound of formula I in which R 9 has one of the above meanings , except hydroxy and amino, and/or R stands for lower alkoxycarbonyl or a ryllavereal oxycarbonyl and the other substituents have the meanings indicated above, esterifies a compound of formula VII, in which at least one of the residues R"^ <a> and R" <*> " <3> stands for carboxy and the other of the residues R"^a and R"*"3 has the above-mentioned 10 9 significance for R respectively for the group R -C(=0)-, 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 the groups which must participate in the reaction, are, if necessary, protected with light cleavable protecting groups, or a reactive carboxylic acid derivative thereof, and then if necessary cleaves off protecting groups present, or e) in a compound of formula I, in which at least one of the residues R8, C(=0)-R9, R10, R11 and C(=0 )-R12 exists in a protected form that does not correspond to the definition of the desired final substance, cleaves off the relevant protecting groups, or f) for the production of a compound of formula I, in which R 9 stands for amino and the other substituents have the meanings indicated above, amidates a compound of formula VIII, in which the residue R 14 stands for carboxy and the other substituents have the above-mentioned meanings, whereby in a compound of formula VIII present free functional groups, with the exception of the groups which will participate in the reaction, can, if necessary, be protected with easily cleavable protective groups, or a reactive carboxylic acid derivative thereof and, if necessary, cleaves off the protective groups present, and, if desired, after carrying out one of the methods a-f) converts a prepared compound of formula I with at least one salt-forming group into its salt, or an obtained salt of a compound of formula I to the free compound, and/or separates an isomer mixture obtained. 2. Process according to claim 1, characterized in that the starting materials are selected in such a way that a compound of formula I is produced in which the exo part is derived from D-glucose, D-mannose or D-galactose, n stands for 0 or 1, and R 1 , R 4 and R stand independently of each other for lower alkanoyl or benzoyl, R <2> stands for lower alkyl or phenyl, R <3> , R <5> and R <7> stand for <r> independently of each other for hydrogen or lower alkyl, or R <5> and R 8 together represent trimethylene and R 7 represents hydrogen, R <8> represents hydrogen or unsubstituted or lower alkyl substituted with phenyl, hydroxy, mercapto or lower alkylthio, R 9 and R 12 independently represent hydroxy, amino, C1 _a .Q -alkoxy, aryl lavereal oxy, alkanoyloxylavereal oxy with up to 16 C atoms, aroyloxylavereal oxy or 3-cholesteryloxy, R10 stands for hydrogen, carboxy, lower alkoxycarbonyl or aryl lavereal oxycarbonyl and R^"*" stands for hydrogen, or unsubstituted or with amino or hydroxy substituted lower alkyl, 9 12 under the condition that at least one of the residues R and R is different from hydroxy, amino or lower alkoxy, or R"^ is different from hydrogen, carboxy or lower alkoxycarbonyl, or a salt of such a compound with at least one salt-forming group. 3. Method according to claim 1 or 2, characterized in that the starting materials are selected in such a way that a compound 9 12 is produced Else of formula I in which R and R independently of each other stand for hydroxy, amino, C1_1Q - alkoxy, alkanoyloxylaeraloxy with up to 16 C atoms, 3-cholesteryloxy or unsubstituted in the phenyl part or with lower alkyl, phenyl, halogen, hydroxy, lower alkoxy, lower alkanoyloxy . di-lower alkylamino or lower alkanoylamino substituted phenyl lower alkyl oxycarbonyl, and the other substituents have the meanings given above, under the condition that at least one of the residues R and R is for different from hydroxy, amino or lower alkoxy, or R 1 is different from hydrogen, carboxy and lower alkoxycarbonyl, or a salt of such a compound with at least one salt-forming group. 4. Process according to claim 1, characterized in that the 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 or 1, R , R and R stand independently for C-.-alkanoyl or benzoyl, R 2 stands for C,.-alkyl or phenyl, R , R and R stand independently for hydrogen or methyl, or R and R together stand for trimethylene and R 7 stands for hydrogen, R 8 for hydrogen, C 1 -alkyl, or with phenyl, hydroxy, mercapto or methylthio substituted C 1 -2 -alkyl, R 9 and R 12 stand independently of each other for hydroxy, amino, lower alkoxy, phenyl-lower-alkoxy, lower-alkanoyloxy-lower-alkoxy, bnezoyloxy-lower-alkoxy or 3-cholesteryloxy, R^ represents hydrogen, carboxy, lower-alkoxycarbonyl or phenyl-lower-alkoxycarbonyl and R"^ stands for hydrogen,.or unsubstituted or with amino or hydroxy substituted C,.-alkyl, under 9 12 the condition that at least one of the residues R and R is different from hydroxy, amino and lower alkoxy, or R is different from hydrogen, carboxy and lower alkoxycarbonyl, or a salt of such a compound with at least one salt-forming group. 5. Method according to claim 1, characterized in that the 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 or 1, R"*" R and R stand independently of each other for C 1 -alkanoyl or benzoyl, R 2 stands for C 1 -alkyl or phenyl, R 3 , R 5 and R 7 stand independently of each other for hydrogen or methyl, R 8 stands for for C 1 -alkyl, R and R 12 independently stand for hydroxy, amino, C 1 . -Alkoxy, phenylmethoxy, lower alkanoyloxymethoxy, benzoyloxymefcoxy or 3-cholesteryloxy, R <10> stands for hydrogen, carboxy, alkoxycarbonyl with up to 5 carbon atoms or phenylmethoxycarbonyl and R stands for C, .-alkyl, on the condition that at least one of the residues R<9> and R 12 are different from hydroxy, amino and C 1 - alkoxy or R 10 is different from hydrogen, carboxy and alkoxycarbonyl with up to 5 C atoms, or a salt of such a compound with at least one salt-forming group. 6. Process according to claim 1, characterized in that the 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 0 or 1, R , R and R stand for acetyl or butyryl , R 2 stands for C, --alkyl or phenyl, R <3> stands for hydrogen or methyl, R <5> and R <7> stand for hydrogen, R <8> stands for C,.,-alkyl, R 9 represents amino, C₁ _4 -alkoxy, pivalyloxymethoxy or mono- or di-phenylmethoxy, R,_^ stands for hydrogen, R 11 stands for methyl and R <12> stands <o> r for mono- or diphenylmethoxy or 3- cholesteryloxy. 7. Process according to claim 1, characterized in that the 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, n stands for 0 or 1, R , R and R stand for C__ ,-alkanoyl, R stands for C. -alkyl or phenyl, R 3 , R 5 and R<7> independently stand for hydrogen or methyl, R 8 stands for C, alkyl, R 9 stands for amino, lower alkoxy, pivaloyloxymethoxy , diphenylmethoxy, benzyloxy or 2-trimethylammonioethoxy, R"^ stands for hydrogen or lower alkoxycarbonyl, R^ stands for C, .-alkyl, lower alkanoyloxymethyl or (2-benzyloxycarbonylamino-ethyl)-sulfonyl-methyl and R 12 stands for amino, lower alkoxy, pivalyloxymethoxy, diphenylmethoxy, benzyloxy, 2-trimethylammonio -ethoxy, 3-cholesteryloxy or benzoyloxymethoxy, provided that 9 12 at least one of the residues R and R is different from amino and lower alkoxy, or a salt of such a compound which is capable of salt formation. 8. Process according to claim 1 or 7, characterized in that the 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 <12> stands for pivaloyloxymethoxy, diphenylmethoxy, benzyloxy, 2-trimethylammonioethoxy, 3-cholesteryloxy or benzoyloxymethoxy and the 9 12 other of the residues R and R have the meaning indicated above, or a salt of such a compound which is capable of salt formation. 9. Process according to claim 1, characterized in that the starting materials are selected in such a way that 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramy1-L-alany1-D-isoglutamine-benzhydry1-ester is produced. 10. Process according to claim 1, characterized in that the starting materials are selected in such a way that 1,4,6-tri-O-acetyl-N-benzoyl-desmethylmuramy1-L-alany1-D-isoglutamine-benzhydryl ester is produced.