DD247606A5 - ACYLATED HEXOSEDERIVATES AND METHOD FOR THE PRODUCTION THEREOF - Google Patents

Abstract

Compounds of the formula I in which the hexose part is derived from D-glucose, D-mannose or D-galactose, n is 0 or 1, and R 1 are lower alkanoyl or benzoyl, R 2 is lower alkyl or phenyl, R 3, R 5 and R 7 are each independent of one another Hydrogen or lower alkyl, or R5 with R8 together trimethylene and R7 hydrogen, R4 and R6 independently hydrogen, lower alkanoyl or benzoyl, R8 is hydrogen or unsubstituted or substituted by phenyl, hydroxy, mercapto or lower alkylthio lower alkyl, R9 and R12 independently of one another hydroxy, amino or Lower alkoxy, R10 is hydrogen, carboxy or lower alkoxycarbonyl and R11 is hydrogen or unsubstituted or by amino, lower alkanoylamino, hydroxy, guanidino, lower alkanoyloxy, 2-benzyloxycarbonylaminoethyl-sulfinyl, 2-benzyloxycarbonylamino-ethyl-sulfonyl, 2-lower alkoxycarbonylamino-ethyl-sulfinyl or 2-lower alkoxycarbonylamino -ethyl-sulfonyl substituted lower alkyl, with the proviso that ss in the compounds in which the pyranose part is derived from D-glucose and at the same time n is 0, R5, R7 and R10 are hydrogen, R8 is methyl, R9 is amino and R12 is hydroxy and in which the radicals R1, R4 and R6 are all three are the same, R1, R4 and R6 are different from acetyl and butyryl when R2 is phenyl and R3 is hydrogen and that R1, R4 and R6 are different from acetyl when R2 and R3 are both methyl, and salts of such compounds having at least one salt-forming group having antiviral activity in vivo and processes for their production. DD # AP

Description

-Λ-

Process for the preparation of acylated hexose derivatives Field of application of the invention

The invention relates to a process for the preparation of 1-O-acylated muramylpeptide and analogous D-mannose or D-galactose derivatives _for pharmaceutical preparations containing these derivatives * and their use as medicaments *

Characteristic of the known technical solutions

French Patent Application No. 74 22 909, publication number 2 292 486, discloses muramyl peptides of the type of the compound N-acetyl-muramyl-L-alanyl-D-isoglutamine ("MDP"). These compounds are described as immunological adjuvants, i. they can be used in combination with vaccines to improve the vaccination success. Own investigations, z. 8. in influenza virus-infected mice have shown that N-acetylmuramyl-L-alanyl-D-isoglutamine per se, i. H. without admixture of vaccines, in the prophylaxis and therapy of viral infections is ineffective. Also, the utility of structurally simple muramyl peptides and their analogs against viral infections has not previously been described.

Object of the invention

The object of the invention is to provide structurally relatively simple and therefore relatively easy

^ -7 / / "> η ο

Murarayl peptide derivatives which are highly effective in the prophylaxis and therapy of viral infections after administration to warm-blooded animals.

Explanation of the essence of the invention

Oer invention has for its object to find new muramyl peptide derivatives having the desired properties and processes for their preparation.

According to the invention, compounds of the formula I

R ³ -CH (0) _{r 9 o}

7 X

r 'O C R

1 II II CNCC-NH-CH-CH ₀ -CH-C-

OR ³ R ^ö (O) Q

NH-CH-C H

in which the hexose part is derived from D-glucose, D-mannose or O-galactose, η is O or 1, and R is lower alkanoyl or benzoyl, R is lower alkyl or

3 5 7 phenyl, R, R and R are independently hydrogen

5 8 7

or lower alkyl, or ⁴ R with R together triraethyl and R

Hydrogen, R and R independently of one another hydrogen,

8 is lower alkanoyl or benzoyl, R is hydrogen or unsubstituted or substituted by phenyl, hydroxy, mercapto or lower alkylthio.

lower alkyl, R ⁹ and R ¹² independently of one another are hydroxy, amino or lower alkoxy, R ^{10 is} hydrogen, carboxy or lower alkoxycarbonyl and R ^{11 is} hydrogen or unsubstituted or by amino, lower alkanoylamino, hydroxy, guanidino, lower alkanoyloxy, 2-benzyloxycarbonylaminoethylsulfinyl, 2-benzyloxycarbonylaminoethyl sulfonyl, 2-lower alkoxycarbonylamino-ethyl-sulfinyl or 2-lower alkoxycarbonylamino-ethyl-sulfonyl substituted lower alkyl, with the hate that, in the compounds in which the fyranose part is derived from D-glucose and at the same time η is 0, R ⁸ , R ⁷ and R ^{10 are} hydrogen, R ^{8 is} methyl, R ^{9 is} amino and R ^{12 is} hydroxy and in which the radicals R ¹ , R ⁴ and R ⁶ all have the same meaning, R ¹ , R * and R ⁶ of Acetyl and butyryl are different when R ^{2 is} phenyl and at the same time R ^{3 is} hydrogen, and that R ¹ , R ¹ * and R ⁶ are different from acetyl, when R ² and R ^{3 are} both methyl, and salts of such compounds having at least one salt-forming group.

The above specification includes N-benzoyl-l, 4,6-tri-O-butyryl-demethylmuramyl-L-alanyl-D-isoglutamine, N-benzoyl-1,4,6-tri-O-acetyl-desmethylrauramyl-L-alanyl- D-isoglutamine and N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-alanyl-D-isoglutamine from the compounds of the formula I.

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

In the case of asymmetric substitution, the configuration at the atoms is CR ³ , CR ⁸ or C-CO-R ⁹ (D), (L) and (D), respectively, as indicated in formula I. The configuration at CR ¹¹ is in the case of asymmetric substitution (L) or (D), preferably (L).

Lower alkanoyl R ¹ , R * and R ⁶ is especially Ca-e-alkanoyl, eg n-hexanoyl, mainly C2-s-alkanoyl and preferably Ca-l ₄ -alkanoyl, for example propionyl, butyryl or preferably acetyl.

Lower alkyl R ² is preferably Ci-i »-, in particular Cj.-2-alkyl.

Lower alkyl R ³ , R ⁵ or R ⁷ is preferably C 1-3 -alkyl, in particular methyl.

Unsubstituted lower alkyl R ⁸ or R ¹¹ is preferably Ci-i »alkyl, for example ethyl, isopropyl, 2-methylpropyl, sec-butyl or especially methyl.

Substituted by phenyl, hydroxy, mercapto or lower alkylthio, in particular methylthio, lower alkyl R ⁸ is preferably correspondingly substituted Ci-2-alkyl, for example benzyl, hydroxymethyl, 1-hydroxy-ethyl, mercaptomethyl or 2-methylthio-ethyl.

Lower alkoxy R ⁹ or R ¹² is preferably C 1-4 -alkoxy, for example methoxy, n-butyloxy or tert-butyloxy.

Lower alkoxycarbonyl R ¹⁰ is preferably alkoxycarbonyl having up to 5 C atoms, for example methoxycarbonyl, n-butyloxycarbonyl or tert. Butyloxycarbonyl.

By hydroxy or lower alkanoyloxy substituted lower alkyl R ¹¹ is in particular correspondingly substituted Cj.-2-alkyl, for example hydroxymethyl, 1-hydroxy-ethyl, lower alkanoyloxymethyl or 1-lower alkanoyloxy-ethyl.

Lower alkyl R ¹ * substituted by amino or lower alkanoylamino is preferably 4-amino-n-butyl or 4-lower alkanoylamino-n-butyl.

Guanidino-substituted lower alkyl R ¹ * is preferably 3-guanidino-n-propyl.

Lower alkyl R ¹¹ substituted by 2-benzyloxycarbonylamino-ethylsulfinyl or -sulfonyl, or by 2-lower-alkoxycarbonylamino-ethylsulfinyl or -sulfonyl is preferably correspondingly substituted methyl, for example C 6 H ₅ -CH 2 -OC (»O) -NH-CH 2 - CH ₂ -S ("0) -CH ₂ -.

The general terms used above and below preferably have the following meanings:

The prefix "lower" denotes radicals up to and including 7, in particular up to and including 4, carbon atoms.

Halogen is especially chlorine or bromine, furthermore fluorine or iodine.

Salifying groups in a compound of formula I are acidic groups, e.g. free carboxyl groups, or basic groups, in particular free amino groups. Depending on the nature of the salt-forming group, the compounds of the formula I form metal or ammonium salts or acid addition salts. Salts of a compound of formula I are preferably pharmaceutically acceptable and non-toxic, e.g. Alkali metal or alkaline earth metal salts, e.g. Sodium, potassium, magnesium or calcium salts, or salts with ammonia or suitable organic amines, wherein primarily aliphatic, cycloaliphatic, cycloaliphatic-aliphatic or araliphatic primary, secondary or tertiary mono-, di- or polyamines, and heterocyclic bases for the salt formation come into question, such as lower alkylamines, eg Triethylamine, hydroxy-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-aminobenzoic acid 2-diethylaminoethyl ester, lower alkylene amines, e.g. 1-ethylpiperidine, cycloalkylamines, e.g. Dicyclohexylamine, or benzylamines, e.g. Ν, Ν'-dibenzylethenediamine, further pyridine-type bases, e.g. Pyridine, collidine or quinoline. Compounds of formula I having at least one basic group may include 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 and lysine. In the presence of several acidic or basic groups, mono or polysalts can be formed. Compounds of formula I with an acidic, e.g. free carboxyl group, and a free basic, e.g. an amino group, may also be in

Form of internal salts «d. H. in zwitterionic form "or one part of the molecule may be present as an inner salt and another part as a normal salt. -

For isolation or purification it is also possible to use pharmaceutically unsuitable salts. For therapeutic use, however, only the pharmaceutically acceptable, non-toxic salts, which are therefore preferred

According to the invention, it has surprisingly been found that the abovementioned compounds of the formula I and their pharmaceutically usable salts are outstandingly suitable both for the prophylaxis and for the therapy of viral infections, as is apparent, for example, from animal experiments, as exemplified in the examples. In these animal studies, animals, such as mice or guinea pigs, are infected with a variety of virus species in a single dose for all or the vast majority of untreated (control) animals

lethal, eg ¹ AjO-Qo ^{1 is infected and the course of} infection is observed in the untreated control animals in comparison to animals treated before, simultaneously with or after the infection with one of the abovementioned compounds or a salt thereof.

It turns out that a prophylactic effect on administration of the compounds of formula I already several days to a few, e.g. four, weeks before infection, and a therapeutic effect even when administered for several days, e.g. 1 week after the infection occurs.

The compounds of the formula I are effective in the abovementioned test on the mouse already in the dose range between 0.0001 mg / kg and 0.1 mg / kg.

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

The compounds of the formula I can be used in particular for the prophylaxis and treatment of diseases which are caused by the viruses specified below [for nomenclature cf. J.L.Melnick, prog. Med. Virol. 26, 214-232 (1980) and 28, 208-221 (1982)]:

Cubic symmetry and naked nucleocapsid DNA viruses, enveloped virion DNA viruses, and cubic and capsid helical symmetry viruses.

The compounds of the formula I are preferably used in the case of DNA viruses with enveloped virion and cubic symmetry of the capsid, in the case of RNA viruses with cubic symmetry of the capsid and naked virion and in the case of RNA viruses with a helical symmetry of the capsid, in which the nucleocapsid sheath is located at the surface membrane, but also in the case of Adenoviridae, Poxvirldae and Coronaviridae, in particular human coronaviruses.

The compounds of the formula I are used in the first place in the case of Herpesviridae, Picornaviridae and Myxoviruses, but also in the case of mastadenoviruses, in particular human adenoviruses, in the case of chordopoxvirinae, such as mainly orthopoxviruses, in particular e.g. Vaccinia viruses, in the case of Reoviridae, mainly (in particular human) rotaviruses, and in the case of Caliciviridae and Rhabdoviridae, such as primarily human vesiculoviruses and horses, cattle and pigs.

Mainly, the compounds of formula I are used in the case of alphaherpesvirinae, such as varicellaviruses, e.g. human varicella zoster viruses, rhinoviruses, cardioviruses and orthomyxoviridae, but also in the case of betaherpesvirinae, in particular human cytomegaloviruses, in the case of aphthoviruses, primarily aupiviruses of ungulates, mainly of cattle, and in the case of paramyxoviridae, as in the first place Line pneumoviruses, eg human respiratory syncytial viruses and, in addition, morbilliviruses or paramyxoviruses, such as parainfluenza viruses, e.g. human parainfluenza viruses, including the Sendai viruses as well as in the case of arboviruses or vesiculoviruses, e.g. Vesicular stomatitis viruses.

First and foremost, the compounds of formula I are used in the case of simplex viruses, e.g. human herpes simplex virus types 1 and 2, in the case of human encephalorayocarditis viruses, in the case of influenza viruses, such as mainly influenza A and influenza B viruses, in the case of vaccinia and parainfluenza viruses, and more particularly in the case of the viruses mentioned in the examples.

The compounds of the formula I can be used for the prophylaxis and therapy of viral infections, in particular in warm-blooded animals including man, by administering them enterally or parenterally, primarily together with suitable auxiliaries or excipients. Preferably, they are applied to the mucosa, e.g. intranasally, rectally, vaginally or on the conjunctiva of the eye,

or orally. However, the antiviral effect also occurs when applied by other routes, e.g. subcutaneously, intravenously, intramuscularly or when applied to normal skin.

The dosage of the active ingredient depends i.a. from the warm-blooded species, the defensive state of the organism, the mode of administration and the type of virus. The dose-response relationship is relatively weak.

For prevention, a single dose of about 0.01 mg to about 10 mg, preferably 0.05 to 1 mg, e.g. 0.2 mg, active ingredient to a warm-blooded animal of about 70 kg body weight, e.g. the people. The prophylactic effect of this dose extends over several weeks. If necessary, e.g. in times of increased risk of infection, one can repeat the administration of this dose.

The therapeutic dose for warm-blooded animals of about 70 kg body weight is between 0.1 mg and 25 mg, preferably between 0.1 and 1 mg, for example at 0.5 mg, especially when administered orally. The dosage in topical, especially intranasal administration is lower by a factor of 10. If necessary, one can repeat the administration of the compounds of formula I _v until the occurrence of an improvement of the disease. Often, however, a single application is sufficient.

The compounds of formula I also possess anti-tumor properties. These are based on their ability, e.g. incorporated in multilamellar liposomes or in phosphate buffered saline (PBS) to activate macrophages in such a way that these endogenous defense cells are able to kill tumor cells (cytotoxicity) or to prevent them from growing (cytostasis). The induction of rat tumoricidal and tumoristatic alveolar macrophages in vitro and in situ can be e.g. show with the following experiment:

Alveolar macrophages are obtained by lung lavage with culture medium. These macrophages are activated either by injection of the test substances into the rats (intravenously or intranasally, in situ activation) or by preincubation for 24 hours with a compound of the formula I in the CO 2 incubator ( in vitro activation). The thus activated macrophages are now incubated with tumor cells for a further 72 hours.

To measure tumoricidal activities of the macrophages, the tumor cells are labeled with ^12S I-iododeoxyuridine before the 72 hour incubation. The non-killed tumor cells, after washing away the radioactivity released by lysed tumor cells, can be measured by residual radioactivity.

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

The compounds of the formula I can thus also be used in the case of warm-blooded animals, including humans, for the therapy of tumor diseases, in particular e.g. to prevent the formation of metastases, e.g. at surgical removal of the primary tumor.

Preference is given to compounds of the formula I in which the hexose part is derived from D-glucose or D-mannose, η is 0, R ¹ , R * and R ⁶ independently of one another are C 9 -s-alkanoyl or benzoyl, R ^{2 is} C 1 - Alkyl or phenyl, R ³ , R ⁵ and R ^{7 are} independently hydrogen or

Methyl, or R ⁵ with R ⁸ - together trimethylene and R ^{7 is} hydrogen, R ^{8 is} hydrogen, Ci-u-alkyl or by phenyl, hydroxy, mercapto or methylthio-substituted Ci-2-alkyl, R ⁹ and R ^{12 are} independently hydroxy, Amino or lower alkoxy, and R ^{10 is} hydrogen, carboxy or lower alkoxycarbonyl, and salts of such compounds having at least one salt-forming group except for N-benzoyl-1,4,6-tri-O-butyryl-demethylmuramyl-L-alanyl-D -isoglutamine, N-benzoyl-1,4,6-tri-O-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine and N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-alanyl -D-isoglutamine and its salts.

Particular preference is given to compounds of the formula I in which the hexose part is derived from D-glucose, η is 1, R ¹ , R * and R ⁶ are each independently C 2-5 -alkanoyl or benzoyl, R ^{2 is} C 1-6 -alkyl or Phenyl, R ³ , R ⁵ and R ⁷ independently of one another are hydrogen or methyl, or R ^{5 together} with R ^{8 is} trimethylene and R ^{7 is} hydrogen, R ^{8 is} hydrogen, C 1-10 -alkyl or C 1 -C 4 -substituted by phenyl, hydroxyl, mercapto or methylthio 2-alkyl, R ⁹ and R ¹² are each independently hydroxy, amino or lower alkoxy, R ^{10 is} hydrogen, carboxy or lower alkoxycarbonyl and R ¹ * is unsubstituted or substituted by amino, guanidino, lower alkanoyloxy, 2-benzyloxycarbonylaminoethylsulfinyl, 2-benzyloxycarbonylaminoethylsulfonyl , 2-lower alkoxycarbonylamino-ethyl-sulfinyl or 2-lower alkoxycarbonylaminoethyl-sulfonyl substituted lower alkyl, and salts of such compounds having at least one salt-forming group.

Very particular preference is given to compounds of the formula I in which the hexose moiety is derived from D-glucose, η is O or 1, and R ^{1 is} C 2-5 -alkanoyl or benzoyl, R ^{2 is} C 1-8 -alkyl or phenyl, R ^{3 is} hydrogen or Ci-i »alkyl, R * is hydrogen, C2 ~ 5-alkanoyl or benzoyl, R ^{5 is} hydrogen, methyl or together with R ^{8 is} trimethylene, R ^{6 is} C2-5-alkanoyl or benzoyl, R ^{7 is} hydrogen or methyl, R ⁸ Ci-i »alkyl or together with R ^{5 is} trimethylene, R ^{9 is} hydroxy, lower alkoxy or amino, R ^{10 is} hydrogen or carboxy, R ^{11 is} unsubstituted or substituted by amino, guanidino or 2-benzyloxycarbonylamino-ethyl-sulfinyl Ci-i» alkyl and R ^{12 is} hydroxy, lower alkoxy or amino

and salts of such compounds having at least one salt-forming group except for N-benzoyl-1,4,6-tri-O-butyryl-demethylmuramyl-L-alanyl-D-isoglutamine, N-benzoyl-1,4,6 tri-O-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine and N-acetyl-1 ^, O-tri-O-acetyl-muramyl-L-alanyl-D-isoglutarain and their salts.

Particular preference is given to compounds of the formula I in which the hexose moiety is derived from D-glucose, η is 0 or 1, R ^{1 is} C 2-5 -alkanoyl or benzoyl, R ^{2 is} C 1-2 -alkyl or phenyl, R ^{3 is} hydrogen or methyl , R * is hydrogen, C 2-5 -alkanoyl or benzoyl, R ^{5 is} hydrogen, R ^{6 is} C 2-5 -alkanoyl or benzoyl, R ^{7 is} hydrogen, R ^{8 is} methyl, ethyl or 2-propyl, R ^{9 is} hydroxy, amino or Ci-1. Alkoxy, R ^{10 is} hydrogen, R ^{11 is} methyl or 4-amino-n-butyl and R ^{12 is} hydroxy, amino of Ci-i * alkoxy, and salts of such compounds having at least one salt-forming group with the exception of N-benzoyl 1,4,6-tri-O-butyryl-demethylmuramyl-L-alanyl-D-isoglutamine, N-benzoyl-1,4,6-tri-O-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine and N- Acetyl-1,4,6-tri-O-acetyl-muramyl-L-alanyl-D-isoglutamine and its salts.

Very particular preference is given to the compounds of the formula I described in the examples.

Very particular preference is also given to the abovementioned compounds of the formula I in which R ¹ is different from acetyl and butyryl and / or R ² is different from methyl and phenyl and / or R * is different from acetyl and butyryl and / or R ^{5 is} hydrogen is different and / or R ⁶ is different from acetyl and butyryl and / or R ^{7 is} different from hydrogen and / or R ^{8 is} different from methyl and / or R? is different from amino and / or R ^{10 is} different from hydrogen and / or η is other than 0 and / or R ^{12 is} other than hydroxy, and salts of such compounds having at least one salt-forming group.

As examples of such compounds, mention may be made of those in which R ^{8 is} C 2-4 -alkyl or, together with R ⁵ , is trimethylene.

Most preferred for their outstanding pharmacological activity are the abovementioned compounds of the formula I in which at least one of the radicals R ⁹ and R ^{12 is} lower alkoxy and the other of the radicals R ⁹ and R ^{12 is} hydroxyl, amino or lower alkoxy, and / or wherein η is 1 and R ¹¹ is other than hydrogen, and the remaining substituents are as defined above, and salts of such compounds having at least one salt-forming group.

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

They are e.g. made by

a) a compound of formula II, CH ₂ OR ^{6 a}

^Λ . UWWdORl (II)

¹ NH-R ²³ R ³ -Cft (D)

-NH-CH-CH ₂

lO

(L)

^Y n

NH-CH-C

R ¹²

wherein at least one of R ¹ , R ² and, for the preparation of compounds of formula I, wherein R ¹ * and R ^{6 have} the abovementioned meanings except for hydrogen, R ¹ * and R ^{60 is} hydrogen, and the remaining of these Groups have the meanings of R ¹ , the group R ² -C (* O) -, R * or R ⁶ , and the remaining substituents have the abovementioned meanings, wherein in a compound of formula II existing free functional groups with the exception Groups to participate in the reaction

if necessary, protected by easily removable protecting groups, with an acyl radical to be introduced R ¹ , R ² -C (»O) -, R * or R ⁶ transmits transferring acylating agents and eliminates any necessary protective groups, or

b) a compound of the formula III, CH ₂ OR ⁶

iH »niwuviiOR ¹ (III)

i- S O

kcR ²

in which the substituents have the abovementioned meanings, where free OH groups OR ¹ * and OR ^{6 are} protected by readily cleavable protective groups, or a reactive derivative thereof with a compound of the formula IV,

η V ° _r r *

X-CH-C-N-C-C-NH-CH-CH ₂ -CH-C-NH-Cl l ö R ⁵ R ⁸ (D) ö

R ¹² (IV)

wherein X is a reactive esterified hydroxy group, and the remaining substituents have the abovementioned meanings, which are protected in a compound of formula IV free functional groups with the exception of X, if necessary, protected by readily cleavable protective groups, reacting and releasing existing protecting groups, if necessary, or

c) a compound of the formula V, H ₂ OR ⁶

* o ~~ ι.

DlO 1

ι Χ 'NH-CH-CH ₂ -CH-C-

(D) ^Ö

(V)

f NH-CH-C-

wherein q, r, s and t are independently O or 1 and wherein the substituents have the abovementioned meanings, wherein in a compound of the formula V existing free functional groups except the group to participate in the reaction, if necessary, by light are cleavable protecting groups protected, or a reactive carboxylic acid derivative thereof with a compound of formula VI,

Ή-CH 2 -CH C-

^ ¹¹

NH-CH-C

wherein u, ν and χ are independently 0 or 1 and the remaining symbols and substituents have the abovementioned meanings, wherein in a compound of formula VI existing free functional groups except for the group to participate in the reaction, if necessary by light cleavable protecting groups are protected, where u, ν and χ are 1, if in the reactant of the formula V q and t are 0, or u are 0 and ν and χ is 1, if q is 1 and t is 0, or u and ν are 0 and χ is 1, when q, r and t are 1 and s is 0 or (for the preparation of compounds of formula I, wherein η is 1

stands) u and χ are O when q, r, s and t are 1, or reacting with a reactive derivative thereof and releasing protective groups present if necessary, or

d) for preparing a compound of the formula I in which R ^{9 has} one of the abovementioned meanings other than hydroxyl, and the other substituents have the abovementioned meanings, a compound of the formula VII

CH ₂ OR ⁶ • -0

ϊ ⁷ 8 Γ Γ

'C-N-C-C-NH-CH-CH ₂ -CH-C-O R ⁵ R ⁸ (D) ö

(D) "OIL ö J

(L)

NH-CH-C

0 η

wherein R ^{13 is} carboxy, and wherein the other substituents have the abovementioned meanings, wherein in a compound of formula VII present free functional groups except R ¹³ are protected if necessary by readily cleavable protecting groups, or a reactive carboxylic acid derivative thereof amidated or esterified and if necessary, splits off existing protective groups, or

e) in a compound of the formula I in which at least one of the radicals OR ", OR ⁶ , R ⁸ , C (O) -R ⁹ , R ¹⁰ , R ¹¹ and C ¹ -O) -R ^{12 is present} in a protected form, which does not correspond to the definition of the desired end product which splits off the corresponding protective group (s),

and, if desired, after carrying out one of the processes a-e), converting a compound of the formula I having at least one salt-forming group into its salt or converting a salt of a compound of the formula I into the free compound, and / or a mixture of isomers obtained separates.

The processes a, c, d and e are preferred.

The implementation of the abovementioned process variants is explained in more detail below:

Method a: Preferably, a) is used to introduce an acyl radical R ¹ , R * and / or R ⁶ used. This is preferably done by Connections de:

R ² -C ("O).

2a Compounds of formula II, wherein the radical R is the group

For the preparation of compounds of formula I wherein R are * and / or R ⁶ is hydrogen, it is preferable to compounds of formula II from wherein R ¹ * ^a and / or R * ^a is an easy and regioselectively cleavable hydroxy-protecting group, it being possible R ⁴ and R ^{6 may} optionally together also represent a bivalent hydroxy protecting group.

In the case of acylating agents containing large space-filling acyl radicals, e.g. Benzoyl transferred, succeeds in a suitable reaction and the selective acylation of the hydroxyl groups in 1- and 6-position of the sugar moiety, without protection of the 4-OH group is mandatory.

In a compound of formula II optionally present free functional groups, which are preferably protected by readily cleavable protecting groups, are also in particular free hydroxy or mercapto in the radical R ⁸ , free carboxy R ⁹ -C (= 0) -, R ¹⁰ or R ^12th -C ("O) - and free amino, hydroxy or guanidino in the radical R ¹¹ .

Protecting groups, their introduction and cleavage are described for example in "Protective Groups in Organic Chemistry", Plenum Press, London, New York 1973, and in "Methods of Organic Chemistry", Houben-Weyl, 4th Edition, Vol. 15/1, Georg-Thieme-Verlag, Stuttgart 1974 and in Theodora W. Greene, "Protective Groups in Organic Synthesis, John Wiley & Sons, New York 1981. It is characteristic of protecting groups that they occur easily, ie without undesirable side reactions, eg solvolytically, can be cleaved reductively, photolytically or under physiological conditions.

Hydroxy protecting groups are e.g. Acyl radicals, as appropriate, e.g. by halogen, substituted lower alkanoyl, such as 2,2-dichloroacetyl, or acyl radicals of carbonic monoesters, especially tert-butyloxycarbonyl, optionally substituted benzyloxycarbonyl, e.g. 4-nitro-benzyloxycarbonyl, or diphenylmethoxycarbonyl, or 2-halo-lower alkoxycarbonyl, such as 2,2,2-trichloroethoxycarbonyl, furthermore trityl or formyl, or organic silyl or stannyl radicals, furthermore easily cleavable etherifying groups, such as tert-lower alkyl, e.g. tert -butyl, 2-oxa or 2-thia-aliphatic or -cycloaliphatic hydrocarbon radicals, primarily 1-lower alkoxy-lower alkyl or 1-lower-alkylthio-lower alkyl, e.g. Methoxymethyl, 1-methoxy-ethyl, 1-ethoxy-ethyl, 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 Thiaanaloge, and optionally substituted 1-phenyl-lower alkyl, such as optionally substituted benzyl or diphenylmethyl, wherein as substituents of the phenyl radicals, for example. Halogen, such as chlorine, lower alkoxy, such as methoxy and / or nitro come into question. Two adjacent hydroxy groups, e.g. the two hydroxy groups in the 4- and 6-position of the sugar moiety can also be replaced by an optionally, e.g. be protected by a phenyl radical, substituted methylene radical or a cycloalkylidene radical, e.g. by lower alkylidene, in particular isopropylidene, or by benzylidene.

'47 606

Carboxyl groups are usually protected in esterified form, such ester groups being readily cleavable under mild conditions. Carboxyl groups protected in this way contain, as esterifying groups, lower-alkyl groups branched in the first position or suitably substituted in the 1- or 2-position. Preferred esterified carboxylic groups include i.a. tertiary lower alkoxycarbonyl, e.g. tert-butyloxycarbonyl, arylmethoxycarbonyl having one or two aryl radicals, these optionally being e.g. by lower alkyl, such as tert-lower alkyl, e.g. tert -butyl, lower alkoxy, such as methoxy, hydroxy, halo, e.g. Chlorine, and / or nitro, mono- or polysubstituted phenyl radicals, as appropriate, 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 alkoxy-lower alkoxycarbonyl such as methoxymethoxycarbonyl, 1-methoxyethoxycarbonyl or 1-ethoxymethoxycarbonyl, 1-lower alkylthio-lower alkoxycarbonyl such as 1-methylthiomethoxycarbonyl or 1-ethylthioethoxycarbonyl, aroylmethoxycarbonyl wherein the aroyl group is optionally substituted, e.g. by halogen, such as bromine, substituted benzoyl, e.g. Phenacyloxycarbonyl, 2-halo-lower alkoxycarbonyl, e.g. 2,2,2-trichloroethoxycarbonyl, 2-bromoethoxycarbonyl or 2-iodoethoxycarbonyl, or 2- (trisubstituted silyl) -ethoxycarbonyl, wherein the substituents each independently represent an optionally substituted, e.g. lower alkyl, lower alkoxy, aryl, halo and / or nitro substituted, aliphatic, araliphatic, cycloaliphatic or aromatic hydrocarbon radical, such as corresponding optionally substituted lower alkyl, phenyl-lower alkyl, cycloalkyl or phenyl, e.g. 2-tri-lower alkylsilylethoxycarbonyl, 2-trimethylsilylethoxycarbonyl or 2- (di-n-butyl-methyl-silyl) -ethoxycarbonyl, or 2-triarylsilylethoxycarbonyl such as 2-triphenylsilylethoxycarbonyl.

The organic silyl or stannyl radicals mentioned above and below preferably contain lower alkyl, in particular methyl, as substituents of the silicon or tin atoms. Corresponding silyl or stannyl groups are primarily tri-lower alkylsilyl, especially trimethylsilyl, further dimethyl-tert-butyl-silyl, or appropriately substituted stannyl, e.g. Tri-n-butylstannyl.

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

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

For example, in a corresponding acylamino group, acyl is the acyl radical of an organic carboxylic acid with e.g. up to 18 carbon atoms, especially an optionally, e.g. by halogen or aryl, substituted alkanecarboxylic acid or optionally, e.g. by halogen, lower alkoxy or nitro, substituted benzoic acid, or a carbonic monoester. Such acyl groups are, for example, lower alkanoyl, such as formyl, acetyl or propionyl, halo-lower alkanoyl, such as 2-haloacetyl, in particular 2-chloro, 2-bromo, 2-iodo, 2,2,2-trifluoro or 2,2,2 Trichloroacetyl, optionally, for example halo, lower alkoxy or nitro substituted benzoyl, e.g. Benzoyl, 4-chlorobenzoyl, 4-methoxybenzoyl or 4-nitrobenzoyl, or lower alkoxycarbonyl branched in the 1-position of the lower alkyl radical or suitably substituted in the 1- or 2-position, in particular tert-lower alkoxycarbonyl, e.g. tert-butyloxycarbonyl, arylmethoxycarbonyl having one or two aryl radicals, preferably optionally, e.g. by lower alkyl, especially tertiary-lower alkyl, such as tert-butyl, lower alkoxy, such as methoxy, hydroxy, halogen, e.g. Chlorine, and / or nitro, mono- or polysubstituted phenyl

247 6Q6

such as optionally substituted benzyloxycarbonyl, e.g. 4-nitro-benzyloxycarbonyl, or substituted diphenylmethoxycarbonyl, e.g. Benzhydryloxycarbonyl or di (4-methoxyphenyl) methoxycarbonyl, aroylmethoxycarbonyl, wherein the aroyl group is preferably optionally, e.g. by halogen, such as bromine, substituted benzoyl, e.g. Phenacyloxycarbonyl, 2-halo-lower alkoxycarbonyl, e.g. 2,2,2-trichloroethoxycarbonyl, 2-bromoethoxycarbonyl or 2-iodoethoxycarbonyl, or 2- (trisubstituted silyl) -ethoxycarbonyl, wherein the substituents each independently represent an optionally substituted, e.g. by lower alkyl, lower alkoxy, aryl, halogen or nitro substituted, aliphatic, araliphatic, cycloaliphatic or aromatic hydrocarbon radical having up to 15 carbon atoms, such as corresponding, optionally substituted lower alkyl, phenyl, alkyl, cycloalkyl or phenyl, e.g. 2-tri-lower alkylsilylethoxycarbonyl such as 2-trimethylsilylethoxycarbonyl or 2- (di-n-butylmethyl-silyl) -ethoxycarbonyl, or 2-triarylsilylethoxycarbonyl such as 2-triphenylsilylethoxycarbonyl.

Other acyl radicals which may be used as amino-protecting groups are also corresponding radicals of organic phosphoric, phosphonic or phosphinic acids, such as diloweralkylphosphoryl, e.g. Dimethylphosphoryl, diethylphosphoryl, di-n-propylphosphoryl or diisopropylphosphoryl, dicycloalkylphosphoryl., E.g. Dicyclohexylphosphoryl, optionally substituted diphenylphosphoryl, e.g. Diphenylphosphoryl, optionally, e.g. nitro-substituted di- (phenyl-lower alkyl) -phosphoryl, e.g. Olibenzylphosphoryl or di- (4-nitrobenzyl) -phosphoryl, optionally substituted phenyloxy-phenyl-phosphonyl, e.g. Phenyloxyphenyl phosphonyl, dilower alkyl phosphinyl, e.g. Oiethylphosphinyl, or optionally substituted diphenylphosphinyl, e.g. Diphenylphosphinyl.

In an arylmethylamino group which represents a mono-, di- or especially triarylmethylamino group, the aryl radicals are in particular optionally substituted phenyl radicals. Such groups are, for example, benzyl, diphenyl methyl and in particular tritylamino.

An etherified mercapto group in an amino group protected with such a radical is primarily arylthio or aryl-lower alkylthio, wherein aryl is especially optionally, e.g. is phenyl substituted by lower alkyl, such as methyl or tert-butyl, lower alkoxy, such as methoxy, halogen, such as chlorine, and / or nitro. A corresponding amino-protecting group is e.g. 4-nitrophenylthio.

In a 2-acyl-lower alk-1-ol-yl radical useful as an amino-protecting group, acyl is e.g. the corresponding residue of a lower alkanecarboxylic acid, an optionally, e.g. by 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 carbonic monoester, such as a carbonic acid lower alkyl. Corresponding protecting groups are primarily 1-lower alkanoyl-prop-1-en-2-yl, e.g. 1-acetyl-prop-1-en-2-yl, or 1-lower alkoxycarbonyl-prop-1-en-2-yl, e.g. 1-ethoxycarbonyl-prop-len-2-yl.

An amino group can also be protected in protonated form; suitable anions are primarily those of strong inorganic acids, such as hydrohalic acids, e.g. the chlorine or bromine anion, or of organic sulfonic acids, such as p-toluenesulfonic acid in question.

Preferred amino protecting groups are acyl radicals of carbonic monoesters, especially tert-butyloxycarbonyl, optionally, e.g. as indicated, substituted benzyloxycarbonyl, e.g. 4-nitro-benzyloxycarbonyl, or diphenylmethoxycarbonyl, or 2-halo-lower alkoxycarbonyl, such as 2,2,2-trichloroethoxycarbonyl, also trityl or formyl.

2 3 T 4 /

A mercapto group, such as in cysteine, can in particular be protected by S-alkylation with optionally substituted alkyl radicals, thioacetal formation, S-acylation or by the preparation of asymmetric disulfide groups. Preferred mercapto protecting groups are, for example, optionally substituted in the phenyl radical, for example by methoxy or nitro benzyl such as 4-methoxybenzyl, optionally in the phenyl moiety, for example by methoxy, substituted diphenylmethyl, such as ^4,4-f -Dimethoxydiphenyl methyl, triphenylmethyl, trimethylsilyl, benzyl-thiomethyl, tetrahydropyranyl, acylaminomethyl, benzoyl, benzyloxycarbonyl or aminocarbonyl, such as ethylaminocarbonyl.

An acylating agent which transfers the radical R ¹ , R ² -C (»0) -, R * or R ⁶ is, in particular, the corresponding carboxylic acid, ie R ¹ -OH, R ² -COOH, R * -OH or R ⁶ -OH, or preferably a reactive acid derivative thereof, wherein the activation of the carboxylic acid used as acylating agent can also be carried out in situ in the presence of the compound of formula II.

Activated carboxylic acid derivatives useful as acylating agents are primarily reactive activated esters or reactive anhydrides, and also reactive cyclic amides.

Activated esters of acids are especially at the linking carbon atom of the esterifying radical unsaturated esters, e.g. vinyl ester type, such as actual vinyl esters (which can be obtained, for example, by transesterification of a corresponding ester with vinyl acetate, activated vinyl ester method), carbamoylvinyl esters (which can be obtained, for example, by treating the corresponding acid with an isoxazolium reagent, 1,2-oxazolium or Woodward method), or 1-lower alkoxyvinyl ester (which can be obtained, for example, by treating the corresponding acid with a lower alkoxyacetylene; ethoxyacetylene method), or amidino-type esters, such as Ν, Ν'-disubstituted amidinoesters (which can be obtained, for example, by treatment the corresponding acid with a suitable Ν, Ν'-disubstituted

Carbodiimide, 2.B. N '' - dicyclohexylcarbodiimide; Carbodiimide method), or Ν, Ν-disubstituted amidinoesters (which can be obtained, for example, by treating the corresponding acid with an N, N-disubstituted cyanamide, cyanamide method), suitable aryl esters, in particular phenyl-substituted by electron-withdrawing substituents ( which, for example, by treating the corresponding acid with a suitably substituted phenol, for example 4-nitrophenol, 4-methylsulfonyl-phenol, 2,4,5-Trichlorpheno'l, 2,3,4,5,6-pentachloro-phenol or 4 Phenyldiazophenol, in the presence of a condensing agent, such as Ν, Ν'-dicyclohexylcarbodiimide, activated aryl ester method), cyanomethyl esters (which can be obtained, for example, by treating the corresponding acid with chloroacetonitrile in the presence of a base, cyanomethyl ester method) , Thioesters, in particular, if appropriate, for example by nitro, substituted phenyl thioesters (which can be obtained, for example, by treating the corresponding acid with thiophenols which are optionally substituted, for example by nitro, inter alia by means of the anhydride or carbodiimide method, activated thiol ester method), amino or amido esters (which, for example, by treating the corresponding acid with an N-hydroxy-amino or N-hydroxy-amido compound, for example N-hydroxy-succinimide, N-hydroxy-piperidine, N-hydroxyphthalimide or 1-hydroxy -benzotriazole, for example by the anhydride or carbodiimide method; the activated N-hydroxy ester method) or silyl esters (which can be obtained, for example, by treating the corresponding acid with a silylating agent, for example hexamethyldisilazane, and which are readily derivable with hydroxyl, but not react with amino groups).

Anhydrides of acids may be symmetrical or preferably mixed anhydrides of these acids, e.g. Anhydrides with inorganic acids, such as acid halides, in particular acid chlorides (which can be obtained, for example, by treating the corresponding acid with thionyl chloride, phosphorus pentachloride or oxalyl chloride, acid chloride method), azides (eg, from a corresponding acid ester over the corresponding hydrazide and its treatment with nitrous acid azide method), anhydrides with carbon

acid half derivatives, as with corresponding esters, e.g. Carbonic acid lower alkyl halides (which are obtained, for example, by treating the corresponding acid with halo, such as lower alkyl chloroformates or with an 1-lower alkoxycarbonyl-2-lower alkoxy-1,2-dihydro-quinoline, for example 1-lower alkoxycarbonyl-2-ethoxy-1, 2- dihydroquinoline), or anhydrides with dihalogenated, in particular dichlorinated, phosphoric acid (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 be obtained, for example, by treating the corresponding acid with an optionally substituted lower alkane or phenylalkanecarboxylic acid halide, for example phenylacetic acid, pivalic acid or trifluoroacetic acid chloride, mixed carboxylic acid anhydride method or with organic sulfonic acids (obtained, for example, by treating a salt, such as an alkali metal salt, the corresponding acid, with a suitable organic sulfonic acid halide, such as lower alkane or aryl, e.g. Methane or p-toluenesulfonyl chloride, can be obtained; Mixed sulfonic acid anhydride method) and symmetrical anhydrides (which can be obtained, for example, by condensation of the corresponding acid in the presence of a carbodiimide or of 1-diethylaminopropyne; symmetrical anhydride method).

Suitable cyclic amides are in particular amides with five-membered diazacycles of aromatic character, such as amides with imidazoles, e.g. Imidazole (which may be obtained, for example, by treating the corresponding acid with Ν, Ν'-carbonyldiimidazole, imidazolide method), or pyrazoles, e.g. 3,5-dimethyl-pyrazole (which can be obtained, for example, via the acid hydrazide by treatment with acetylacetone; pyrazolide method).

As mentioned, derivatives of acids used as acylating agents can also be formed in situ. So you can, for example Form Ν, Ν'-disubstituted amidinoesters in situ by dissolving 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. NjN'-dicyclohexylcarbodiimide. Further, one may form amino or amido esters of the acids used as acylating agent 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 a Ν, Ν'-disubstituted carbodiimides, 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 may be carried out in a manner known per se, the reaction conditions depending primarily on whether and how the carboxyl group of the acylating agent is activated, usually in the presence of a suitable solvent or diluent or a mixture of such and, if necessary, in the presence of a condensation agent which, when participating in the reaction carboxyl group is, for example, as an anhydride, may also be an acid-binding agent, with cooling or heating, for example in a temperature range of about -30 ⁰ C to about +150 ⁰ C, particularly from about 0 ⁰ C to H-IOO ⁰ C, preferably from room temperature (about +20 ⁰ C) to +70 ⁰ C, in a closed reaction vessel and / or in the atmosphere of an inert gas, for example nitrogen. Typical condensing agents are, for example, carbodiimides, for example N, N'-diethyl, Ν, Ν'-dipropyl, Ν, Ν'-dicyclohexyl or N-ethyl-N'- (3-dimethylaminopropyl) -carbodiimide, suitable carbonyl compounds, for example, carbonyldiimidazole, or 1,2-oxazolium compounds, eg, 2-ethyl-5-phenyl-1,2-oxazolium-3'-sulfonate and 2-tert-butyl-5-methylisoxazolium-perchlorate, or a suitable acylamino compound, For example, 2-ethoxy-l-ethoxycarbonyl-l, 2-dihydroquinoline. Typical acid-binding condensing agents are, for example, alkali metal carbonates or bicarbonates, for example sodium or

247 6ζ

Potassium carbonate or bicarbonate (usually together with a sulphate), or organic bases "such as usually sterically hindered tri-lower alkylamines, e.g. Ν, Ν-diisopropyl-N-ethyl-amine.

Cleavage of the protecting groups, e.g. the carboxyl, amino, hydroxy or mercapto protecting groups which are not part of the desired end product of formula I are carried out in a manner known per se, e.g. by solvolysis, in particular hydrolysis, alcoholysis or acidolysis, or by means of reduction, in particular hydrogenolysis or chemical reduction, optionally in stages or simultaneously, it also being possible to use enzymatic methods.

Thus, tert.-lower alkoxycarbonyl or lower alkoxycarbonyl or optionally substituted diphenylmethoxycarbonyl substituted in the 2-position by an organic silyl group or in the 1-position by lower alkoxy or lower alkylthio may be used. 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, in free carboxyl. Optionally substituted benzyloxycarbonyl may be e.g. by means of hydrogenolysis, i. by treatment with hydrogen in the presence of a metallic hydrogenation catalyst, such as a palladium catalyst. Further, suitably substituted benzyloxycarbonyl such as 4-nitrobenzyloxycarbonyl may also be obtained 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 donating agent capable of producing nascent hydrogen with the metal, such as an acid, primarily a suitable carboxylic acid, such as an optional, e.g. by hydroxy, substituted lower alkanecarboxylic acid, e.g. Acetic acid, formic acid, glycolic acid, diphenyl glycolic acid, lactic acid, mandelic acid, 4-chloro-mandelic acid or tartaric acid, or an alcohol or thiol, preferably adding water, convert into free carboxyl. By treating with a reducing

247 6Oi

Metal or metal salt, as described above, one can also convert 2-halo-lower alkoxycarbonyl (optionally after conversion of a 2-bromo-lower alkoxycarbonyl group to a corresponding 2-iodo-lower alkoxycarbonyl group) or aroylmethoxycarbonyl into free carboxyl, wherein aroylmethoxycarbonyl also by treatment with a nucleophilic, preferably salt-forming reagent such as sodium thiophenolate or sodium iodide can be cleaved. Substituted 2-silylethoxycarbonyl may also be obtained by treatment with a fluoride anion-providing salt of fluorohydroic acid, 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 alkylammonium fluoride or tri-lower alkylarylammonium fluoride, e.g. Tetraethylammonium fluoride or tetrabutylammonium fluoride, in the presence of an aprotic polar solvent such as dimethyl sulfoxide or Ν, Ν-dimethylacetamide, are converted into free carboxyl.

Esterified carboxyl can also be cleaved enzymatically, e.g. esterified lysine, e.g. Lysine methyl ester, by trypsin.

A protected amino group is set free in known per se and depending on the nature of the protective groups in various ways, preferably by solvolysis or reduction. 2-halo-lower alkoxycarbonylamino (optionally after conversion of a 2-bromo-lower alkoxycarbonylamino group to a 2-iodo-lower alkoxycarbonylamino group), aroylmethoxycarbonylamino or 4-nitrobenzyloxycarbonylamino can be e.g. by cleavage 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 prepared by treatment with a nucleophilic, preferably salt-forming, reagent, such as sodium thiophenolate, and 4-nitrobenzyloxycarbonylamino also by treatment with an alkali metal, e.g. Sodium dithionite, are split. Optionally substituted diphenylmethoxycarbonylamino, tert.-lower alkoxycarbonylamino or 2-trisubstituted silylethoxycarbonylamino

amino can be removed by treatment with a suitable acid, e.g. Formic or trifluoroacetic acid, optionally substituted benzyloxycarbonylamino e.g. by means of hydrogenolysis, i. by treating with hydrogen in the presence of a suitable hydrogenation catalyst such as a palladium catalyst, optionally substituted triarylmethylamino or formylamino e.g. by treating with an acid such as mineral acid, e.g. Hydrochloric acid, or an organic acid, e.g. Formic, acetic or trifluoroacetic acid, optionally in the presence of water, and an amino group protected with an organic silyl group may e.g. be released by hydrolysis or alcoholysis. One by 2-haloacetyl, e.g. 2-chloroacetyl, protected amino group may be released by treatment with thiourea in the presence of a base, or with a thiolate salt such as an alkali metal thiolate, the thioureas and subsequent solvolysis such as alcoholysis or hydrolysis of the resulting condensation product. A 2-substituted silylethoxycarbonyl-protected amino group may also be converted to the free amino group by treatment with a fluoride anion-providing salt of fluorohydroic acid, as described above in connection with the release of a suitably protected carboxyl group.

Amino protected in the form of an azido group is converted, for example, by reduction into 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 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, for example methylene chloride, or else in water or a mixture of water and an organic solvent such as an alcohol or dioxane, at about 20 ⁰ C to 25 ° C, or under cooling or heating.

. ". 247 6

A hydroxy or mercapto group protected by a suitable acyl group, an organic silyl group or by optionally substituted 1-phenyl-lower alkyl is liberated analogously to a correspondingly protected amino group. By optionally substituted 1-phenyl-lower alkyl, e.g. Benzyl, protected hydroxy is preferably prepared by catalytic hydrogenation, e.g. in the presence of a palladium-carbon catalyst, cleaved. A 2,2-dichloroacetyl protected hydroxy or mercapto group is e.g. by basic hydrolysis, a hydroxy or mercapto group etherified by tert-lower alkyl or by a 2-oxa or 2-thia-aliphatic or -cycloaliphatic hydrocarbon radical by acidolysis, e.g. by treatment with a mineral acid or a strong carboxylic acid, e.g. Trifluoroacetic acid, released. Two hydroxy groups taken together by means of a preferably substituted methylene group, such as lower alkylidene, e.g. Isopropylidene, cycloalkylidene, e.g. Cyclohexylidene, or benzylidene, can be released by acid solvolysis, especially in the presence of a mineral acid or a strong organic acid. With an organic silyl group, e.g. Trimethylsilyl, etherified hydroxy may also be reacted with a fluoride anion supplying salt of hydrofluoric acid, e.g. Tetrabutylammonium fluoride, are released.

Preferably, in the presence of multiple protected functional groups, the protecting groups are selected to simultaneously cleave more than one such group, for example acidolytically, such as by treatment with trifluoroacetic acid or formic acid, or reductive, such as by treatment with zinc and acetic acid, or with hydrogen and a hydrogenation catalyst such as a palladium-carbon catalyst. When the desired end-products contain protecting groups, for example when R ¹¹ is lower-alkyl substituted by 2-benzyloxycarbonylamino-ethyl-sulfinyl, those protecting groups which are to be split off after the reaction have been selected so that they can be cleaved off regioselectively, eg etherified ethereal silyl group

* ♦ / O I

Hydroxy in the radical R *, R * or R ^{8 are released} with fluoride, wherein a benzyloxycarbonyl protecting group is retained elsewhere in the molecule.

The starting material of the formula II in which R ^{2 is} hydrogen is obtained, for example, from corresponding compounds in which R ^{2 is} an easily cleavable amino-protecting group, for example benzyloxycarbonyl, by removal of this amino-protecting group, for example benzyloxycarbonyl by hydrogenation in the presence of a catalyst consisting of 10% palladium on carbon, can be cleaved off.

Method b:

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

Reactive esterified hydroxy X is e.g. hydroxy esterified with a strong inorganic or organic acid, such as with a mineral acid, e.g. Hydrohalic acid, such as hydrochloric, hydrobromic or hydriodic acid, also sulfuric acid, or halogenated sulfuric acid, e.g. Fluorosulfuric acid, or with a strong organic sulfonic acid such as an optional, e.g. by halogen, such as fluorine, substituted lower alkanesulfonic acid or an aromatic sulfonic acid, e.g. an optionally substituted by lower alkyl, such as methyl, halogen, such as bromine, and / or nitro substituted benzenesulfonic acid, e.g. a methanesulfonic, trifluoromethanesulfonic or p-toluenesulfonic acid, esterified hydroxy, preferably a chloride, bromide or iodide.

Free functional groups in a compound of the formula IV, which are advantageously protected by readily cleavable protective groups, are in particular hydroxyl, mercapto or carboxy groups.

247 oQ-6

The reaction is preferably carried out in an inert organic solvent at a temperature between -3O ⁰ C and +150 ⁰ C, in particular 0 ⁰ C and + 100 ⁰ C, for example +20 ⁰ C and + 70 ⁰ C, if necessary in the presence of an acid-binding agent carried out.

The cleavage of protective groups which are not part of the desired end product of the formula I is carried out as described in process a).

Method 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 anaerobic manner as in the reactive acylating agents described in method a) and wherein the activation also can be done in situ.

In compounds of formulas V or VI optionally present functional groups, which are preferably protected by readily removable protecting groups, are in particular amino and guanidino in the radical R ¹¹ , carboxy as the radical R ¹⁰ and hydroxy as the radical R ⁹ or R ¹² , but also hydroxy or mercapto in the radical R ⁸ or hydroxy in the radical R ¹¹ .

In a reactive derivative of a compound of formula VI, the amino group is e.g. by reaction with a phosphite, e.g. Diethyl chlorophosphite, 1,1-phenylene chlorophosphite, ethyl dichlorophosphite, ethylene chlorophosphite or tetraethyl pyrophosphite, or by bonding to halocarbonyl, e.g. Chlorocarbonyl, or activated in the form of an isocyanate group. Preferably, the reaction is carried out by reacting a reactive carboxylic acid derivative of a compound of formula V with a compound of formula VI wherein the amino or hydroxy group participating in the reaction is in free form.

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

The esterification of a compound of the formula V in which q, r and t are 1 can also be carried out with the aid of the esterification agents described in process d).

Method d:

Free functional groups present in a compound of the formula VII, which are preferably protected by readily cleavable protective groups, are, in particular, free carboxy R ¹⁰ and -C (O) -R ¹² . If appropriate, free hydroxy OR ⁴ and OR ⁶ , free hydroxy or mercapto in the radical R ⁸ or free amino, hydroxy or guanidino in radical R ^{11 are also} protected. The protecting groups and their cleavage are described in method a).

In particular, a reactive carboxylic acid derivative of a compound of formula VII is a reactive ester, a reactive anhydride or a reactive cyclic amide in which the carboxyl group is activated in a manner analogous to the reactive acylating agents described in method a) and wherein the activation is also in situ can. Preferably, the reaction is carried out by reacting a reactive carboxylic acid derivative of a compound of formula VII with a compound HR? in which R ^{9 has} the abovementioned meanings of R? with the exception of hydroxy.

Alternatively, it is possible to esterify a compound of the formula VII with a free carboxyl group by reaction with a reactive derivative of the alcohol desired as the esterification component.

Suitable esterification agents include, for example, corresponding diazo compounds such as optionally substituted diazo-lower alkanes, e.g. Diazomethane, diazoethane or diazo-n-butane. These reagents are dissolved in the presence of a suitable inert solvent.

- jj - efe "» ί ⁵ V ~ * 0 m

by means of such as an aliphatic, cycloaliphatic or aromatic hydrocarbon, such as hexane, cyclohexane, benzene or toluene, of a halogenated aliphatic hydrocarbon, e.g. Methylene chloride, or an ether such as a di-lower 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, with cooling, at room temperature or with gentle heating, further, if necessary, in a closed vessel and / or under an inert gas, e.g. Nitrogen atmosphere, brought to application.

Other suitable esterification agents 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 hydriodic acid, also sulfuric acid, or halogenosulfuric acid, e.g. Fluorosulfuric acid, or strong organic sulfonic acids, as appropriate, e.g. by halogen, such as fluorine, substituted lower alkanesulphonic acids, or aromatic sulphonic acids, e.g. optionally benzenesulfonic acids substituted by lower alkyl, such as methyl, halo, such as bromo, and / or nitro, e.g. Methanesulfon, trifluoromethanesulfonic or p-toluenesulfonic acid. Such esters are i.a. Lower alkyl halides, di-lower alkyl sulfates such as dimethyl sulfate, further fluorosulfate esters such as lower alkyl esters, e.g. Fluorosulfonic acid methyl ester, or optionally halo-substituted methanesulfonic acid lower alkyl esters, e.g. Trifluoromethanesulfonate. They are usually in the presence of an inert solvent, such as an optionally halogenated, such as chlorinated, aliphatic, cycloaliphatic or aromatic hydrocarbon, e.g. Methylene chloride, an ether such as dioxane or tetrahydrofuran, or a mixture thereof. It is preferable to use suitable condensing agents such as alkali metal carbonates or bicarbonates, e.g. Sodium or potassium carbonate or bicarbonate (usually together with a sulfate), or organic bases such as commonly hindered tri-lower alkylamines, e.g. Ν, Ν-diisopropyl-N-

ethyl-amine (preferably together with halosulfonic acid-lower alkyl or optionally halogen-substituted methanesulfonic lower alkyl), wherein with cooling, at room temperature or with heating, for example at temperatures of about -20 ⁰ C to about 50 ⁰ C, and, if necessary, in a closed Vessel and / or in an inert gas, eg nitrogen atmosphere, is worked.

Further agents for esterification are corresponding trisubstituted oxonium salts (so-called seaweed salts), or disubstituted carbenium or halonium salts in which the substituents are the etherifying radicals, for example tri-lower alkyloxonium salts, and di-lower alkoxycarbenium or di-lower alkyl halonium salts, in particular the corresponding salts with complex fluorine-containing acids, such as the corresponding salts Tetrafluoroborates, hexafluorophosphates, hexafluoroantimonates, or hexachloroantimonates. Such reagents are, for example, trimethyloxonium or triethyloxonium hexafluoroantimonate, hexachloroantimonate, hexafluorophosphate or tetrafluoroborate, dimethoxycarbenium hexafluorophosphate or dimethylbromonium hexafluoroantimonate. It is preferable to use these agents in an inert solvent such as an ether or a halogenated hydrocarbon such as diethyl ether, tetrahydrofuran or methylene chloride, or in a mixture thereof, if necessary, in the presence of a base such as an organic base, for example one, preferably sterically hindered Triniederalkylamins, eg Ν, Ν-diisopropyl-N-ethyl-amine, and with cooling, at room temperature or with gentle heating, for example at about -20 ⁰ C to about 50 ⁰ C, if necessary, in a closed vessel and / or in an inert gas, eg nitrogen atmosphere.

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

247 604

- 35 -

Procedure it

A compound of the formula I in which at least one of the radicals R *, R ⁶ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ and R ^{12 is present} in a protected form which does not correspond to the definition of the desired end product is in particular a compound of the formula I in which a hydroxy group OR * or OR ⁶ , a hydroxy or mercapto group in the radical R ⁸ , a carboxyl group R ¹ °, an amino or hydroxy group in the radical R ¹ * and / or free hydroxy R? or R ^{12 are} protected by a readily cleavable protecting group not included in the desired end product.

Easily removable protective groups are in particular those mentioned in process a). The cleavage of the protective groups is carried out analogously, as described in process a).

Additional operations ; Salts of compounds of the formula I with salt-forming groups can be prepared in a manner known per se. Thus, salts of compounds of formula I with acidic groups by reaction with a suitable base, for example by treatment with suitable metal compounds, such as alkali metal salts of suitable organic carboxylic acids, for example the sodium salt of a-ethyl-caproic acid, or with suitable inorganic alkali or Alkaline earth metal salts, especially those derived from a weak and preferably volatile acid, eg sodium bicarbonate, or with ammonia or a suitable organic amine, preferably using stoichiometric amounts or only a small excess of the salt-forming agent. Acid addition salts of compounds of the formula I are obtained in a customary manner, for example by treatment with an acid or a suitable anion exchange reagent. Internal salts of compounds of the formula I which contain, for example, a free carboxyl group and a free amino group can be formed, for example, by neutralizing salts, such as acid addition salts, to the isoelectric point, for example with weak bases, or by treatment with liquid ion exchangers.

'47 6Q

Salts can be converted to the free compounds in the usual way, metal and ammonium salts e.g. by treatment with suitable acids, and acid addition salts e.g. by treating with a suitable basic agent.

Mixtures of isomers may be prepared in a manner known per se, e.g. fractionated crystallization, chromatography etc. are separated into the individual isomers.

The methods described above, including the methods for deprotection and the additional process measures are, unless otherwise indicated, in a conventional manner, for example in the presence or absence of preferably inert solvents and diluents, if necessary, in the presence of Condensation agents or catalysts, at reduced or elevated temperature, for example in a temperature range from about -20 ⁰ C to about +150 ⁰ C, in particular from about 0 ⁰ C to about + 70 ⁰ C, preferably from about +10 ⁰ C to about + 40 ⁰ C, mainly at room temperature in a closed vessel and / or in an inert gas, such as nitrogen atmosphere performed.

Taking into account all the substituents in the molecule, if necessary, e.g. in the presence of easily hydrolyzable radicals, particularly mild reaction conditions, such as short reaction times, use of mild acidic or basic agents in low concentration, stoichiometric proportions, choice of suitable catalysts, solvents, temperature and / or pressure conditions to apply.

The invention also relates to those embodiments of the process which start from a compound obtainable as an intermediate at any stage of the process and which carry out the missing process steps or which terminate the process at any stage or form a starting material under the reaction conditions or in the form of a reactive one

Derivative or salt used. In this case, it is preferable to start from those starting materials which, according to the method, lead to the compounds described above as being particularly valuable.

Novel starting materials and / or intermediates and processes for their preparation are also the subject of the present invention. Preferably, such starting materials are used and the reaction conditions are chosen so as to arrive at the compounds listed in this application as being particularly preferred.

The invention also relates to pharmaceutical preparations containing an amount of active substance effective for the prophylaxis or therapy of viral infections, together with pharmaceutically acceptable excipients which are useful in the topical, e.g. intranasal, enteral, e.g. oral or rectal, or parenteral administration, and may be inorganic or organic, solid or liquid. Thus, tablets or gelatin capsules containing the active ingredient together with diluents, e.g. Lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycerin, and / or lubricants, e.g. Silica, talc, stearic acid or salts thereof, such as magnesium or calcium stearate, and / or polyethylene glycol / tablets may also be binders, e.g. Magnesium aluminum silicate, starches such as corn, wheat or rice starch, gelatin, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidone, and, if desired, disintegrants, e.g. Strong, agar, alginic acid or a salt thereof, such as sodium alginate, and / or effervescent mixtures, or adsorbents, dyes, flavorings and sweeteners. Further, the pharmacologically active compounds of the present invention may be used in the form of parenterally administrable preparations or infusion solutions. Such solutions are preferably isotonic aqueous solutions or suspensions, e.g. for lyophilized preparations containing the active ingredient alone or together with a carrier material, e.g. Mannitol, can be prepared before use. The pharmaceutical preparations may be sterilized and / or adjuvants, e.g. preservatives,

2 <* -KSf A 4 / Q - 38 -

Stabilizers, wetting agents and / or emulsifiers, solubilizers, salts for regulating the osmotic pressure and / or buffers. The present pharmaceutical compositions which, if desired, may contain other pharmacologically active substances, such as antibiotics, are prepared in a manner known per se, e.g. by means of conventional mixing, granulating, coating, dissolving or lyophilization processes, and containing from about 0.001% to 20%, in particular from about 0.01% to about 10%, primarily between 0.1% and 5% , of the active substance (s), an active substance concentration of less than 1% being particularly suitable for topically applied preparations.

Preferred topically applied dosage forms are the following: creams, ointments or pastes having an active ingredient content of 0.001% to 1%, in particular 0.01% to 0.1%, e.g. 0.05%, e.g. Ointments for intranasal administration or lipsticks, or aqueous solutions having an active ingredient content of 0.001% to 1%, in particular 0.05% to 0.5%, e.g. 0.1%, preferably isotonic, sterile and physiologically acceptable solutions, e.g. Eye drops, preferably in single-use micro-containers or sprays for use in the mouth and throat.

Particularly suitable are the pharmaceutical preparations described in the examples.

Creams are oil-in-water emulsions containing more than 50% water. As an oily basis, fatty alcohols, e.g. Lauryl, cetyl or stearyl alcohols, fatty acids, e.g. Palmitic or stearic acid, liquid to solid waxes, e.g. Isopropyl myristate, wool wax or beeswax, and / or hydrocarbons, e.g. Vaseline (petrolatum) or paraffin oil. Suitable emulsifiers are surface-active substances with predominantly hydrophilic properties, such as corresponding nonionic emulsifiers, e.g. Fatty acid esters of polyalcohols or ethylene oxide adducts thereof, such as polyglycerol fatty acid esters or polyoxyethylene sorbitan fatty acid ethers (tweens), furthermore polyoxyethylene fatty alcohol

7 OQi

ether or fatty acid esters, or corresponding ionic emulsifiers, such as alkali metal salts of fatty alcohol sulfates, e.g. Sodium lauryl sulphate, sodium cetyl sulphate or sodium stearyl sulphate, usually in the presence of fatty alcohols, e.g. Cetyl alcohol or stearyl alcohol. Additives to the water phase are u.a. Agents which reduce the drying of the creams, e.g. Polyalcohol, such as glycerol, sorbitol, propylene glycol and / or polyethylene glycols, preservatives, fragrances, etc.

Ointments are water-in-oil emulsions containing up to 70%, but preferably from about 20% to about 50% water or aqueous phase. The fatty phase is primarily hydrocarbons, e.g. Vaseline, paraffin oil and / or hard paraffins in question, the preferably water-binding capacity suitable hydroxy compounds, such as fatty alcohols or esters thereof, e.g. Cetyl alcohol or wool wax alcohols, or wool wax included. Emulsifiers are corresponding 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. Humectants, such as polyalcohols, e.g. Glycerol, propylene glycol, sorbitol and / or polyethylene glycol, such as preservatives, fragrances, etc.

Fatty ointments are anhydrous and contain, in particular, hydrocarbons, e.g. Paraffin, vaseline and / or liquid paraffins, further natural or partially synthetic fats, e.g. Coconut fatty acid triglyceride, or preferably hardened oils, e.g. hydrogenated peanut or castor oil, also fatty acid partial esters of glycerol, e.g. Glycerol mono- and distearate, as well as e.g. mentioned in connection with the ointments, the hydrogen absorbing capacity increasing fatty alcohols, emulsifiers and / or 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 to bind existing moisture or secretions.

Foams are administered from pressure vessels and are aerosolized liquid oil-in-water emulsions wherein halogenated hydrocarbons such as chlorofluoro-lower alkanes, e.g. As dichlorodifluoromethane and dichlorotetrafluoroethane, can be used as a blowing agent. As the oil phase used u. a. Hydrocarbons, eg. As paraffin oil, fatty alcohols, eg. B. CetylalkohA, fatty acid esters, eg. B. Isopropylmy ristalt, and / or other waxes. As emulsifiers u u. a. Mixtures of those having predominantly hydrophilic properties, such as polyoxyethylene sorbitan-fensäureester (Tweens), and those having predominantly lipophilic properties, such as sorbitan fatty acid ester (Spans). There are also the usual additives, such as preservatives, etc.

Tinctures and solutions usually have an aqueous-ethanolic basis, the u. a. Polyalcohols, for. As glycerol, glycols and / or polyethylene glycol, as a humectant to reduce ^ evaporation, and lubricating substances / such as fatty acid esters with low polyethylene glycols, d. H. soluble in aqueous mixture, lipophilic substances as a substitute for the skin with the ethanol extracted fatty substances, and, if necessary, other auxiliaries and additives are added.

The preparation of the topically usable pharmaceutical preparations is carried out in a conventional manner, for. By dissolving or suspending the active ingredient in the base or in a part thereof, if necessary. When the active ingredient is processed as a solution, it is usually dissolved in one of the two phases before emulsification; bsi

- 40a -

Processing as a suspension, it is mixed after emulsification with part of the base and then added to the rest of the formulation.

Embodiment .

The following examples illustrate the invention without limiting it in any way. The R 1 values are determined on silica gel thin layer plates (Merck, Darmstadt, Germany). The ratio of the eluents in the solvent mixtures used in each other in volume fractions (V / V), temperatures are in degrees

- 41 -

Celsius indicated. The concentration, c, of the substance in the solvent (mixture) is given in the case of optical rotation in percent (weight / volume).

Abbreviations ;

Section. * absolutely

Boc tert.Butyloxycarbonyl HV «high vacuum

i.Vak. »in vacuum

Me - methyl

MeOH «methanol PTFE - polytetrafluoroethylene, Teflon · RT * room temperature M.p. = * melting point Dec. "Decomposition

Example 1 ; A solution of 5.77 g (7.3 mmol) of 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester (o, g-mixture) in 80 ml Methanol tetrahydrofuran (1: 1) is hydrogenated for 1 hour at room temperature and atmospheric pressure with 311 mg of 10% palladium carbon catalyst. Then, the catalyst is filtered off and the filtrate is evaporated in vacuo at 30 ° to dryness. The resulting yellow residue is distributed three times between water-saturated n-butanol and water. The Butanolphasen are combined and evaporated in vacuo at 40 °. The resulting, almost colorless residue is dissolved in 100 ml of double-distilled water-tert-butanol (1: 1) and the resulting solution is filtered through a Millipore filter (Fluoropore®, PTFE, 0.2 μm) Filtrate lyophilized under high vacuum.

Pure ^, δ-tri-O-acetyl-N-propionyl-desmethylmuramyl-L is obtained.

alanyl-D-isoglutamine (α, β mixture) as a colorless powder containing 1.16 moles of water and 0.437 moles of t-butanol.

C ₂₅ H ₃₈ N ^ On, * !, 16 H ₂ OO, 437 _(CH3) 3C-OH (671.87) Ber. C, 47.82; H, 6.70; N, 8.34; 37.14: H ₂ O: 3.27 (CH ₃ ) ₃ C-OH, 4.82

- 42 -

Found C 47.70 H 6.61 N 8.51 O 36.99 H ₂ O 3.27 (CH ₃ ) ₃ C-OH 4.82

on [α] ρ - +39.0 ± 0.1 ° (c - 0.682, methanol),

R_ 0.51 (methylene chloride: methanol: water 70: 30: 5). The starting material is obtained as follows:

Step 1.1: 16.0 g (31.63 mmol) of N-propionyl-demethylmuramyl-L-alanyl-D-isoglutamine containing 0.74 mol of water are dissolved in 300 ml of methanol-dimethoxyethane (1: 1) and then 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 diphenyldiazomethane are added in each case. After completion of the reaction (90 hours), the red solution is evaporated to dryness in vacuo at 40 °, the resulting red resin with 300 ml of absolute diethyl ether and stirred for 1/2 hour at room temperature.

This gives colorless crystals, which are suction filtered and washed with diethyl ether.

After recrystallization from 400 ml of acetone to give N-propionyl-desmethylmuramyl-L-alanine-D-isoglutamine-benzhydryl ester (α, β mixture) in the form of colorless crystals of mp. 188-190 ° (dec.), The 1.17 Mol contain water.

· 1.17 H ₂ O (679.78) Ber. C 56.55 H 6.60 N 8.24 0 28.63 H ₂ O 3.09 Gef. C 56.67 H 6.67 N 8.43 O 28.65 H ₂ O 3.09 [<*] £ - + 7.5 + 0.1 ° (c - 0.898; methanol), R - 0.62 (chloroform: methanol: water - 70: 30: 5), R - 0.62 (chloroform: methanol - 7: 3).

Step 1.2: 11.4 g (16.77 mmol) of N-propionyl-demethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester (a, 8 mixture) containing 1.17 mol of water are dissolved in 120 ml of absolute pyridine solved. The way

247

- 43 -

7.6 ml (80.3 mmol) of acetic anhydride are added to the solution obtained and the mixture is stirred at room temperature for 92 hours. Thereafter, the yellowish solution is evaporated in a high vacuum at 40 °. The crude product thus obtained is triturated with 100 ml of diethyl ether and the resulting suspension is stirred for 2 hours at room temperature. Then, the crystals formed are filtered off with suction and washed with diethyl ether.

After two recrystallizations from ethyl acetate-isopropanol-diethyl ether (100: 5: 40) to obtain a first fraction of chromatographically pure 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamin- Benzhydrylester (α, β mixture) in the form of colorless crystals, mp. 165-166 °.

The mother liquors of the recrystallizations are combined and evaporated in vacuo. The residue thus obtained is purified by column chromatography on 600 g of silica gel (type 60, very pure, Merck, 0.063-0.2 mm) in the system chloroform-ethanol (9: 1) (10 ml fractions).

Fractions 217-310 are combined and evaporated in vacuo. This gives a second fraction of chromatographically pure 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester (α, β-mixture), which together with the 1st fraction is dissolved in 200 ml of absolute methanol. The slightly turbid solution thus obtained is then filtered through a Millipore filter (Fluoropore, PTFE, 0.2 μm) and the clear filtrate is evaporated in vacuo at 40 °. The residue is then dissolved in 50 ml of absolute ethyl acetate, which has been previously filtered through a Millipore filter (Fluoropore, PTFE, 0.2 μΐη), and crystallized by addition of 20 ml of absolute diethyl ether, which has also been filtered through a Millipore filter, and washed with filtered, absolute diethyl ether. This gives 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester (α, β mixture) in the form of colorless crystals of mp. 165-166 °, which still Contain 0.27 mol of water.

- 44 - 24

· 0.27H ₂ O (789.68) Ber. C 57,80 H 6,22 N 7,10 0 28,91 H ₂ O 0,62 Gef. C 57,91 H 6,20 N 7,11 O 28,77 H ₂ O 0,62

[α] ρ ° - + 32.7 t 2.1 ° (c - 0.486, methanol), R - 0.28 (chloroform: methanol 9: 1), R_ »0.70 (chloroform! methanol 4: 1) ,

Another product is obtained from fractions 105-140 of the above-described column chromatography.

The yellow residue obtained after evaporation in vacuo (0.55 g, foam) is dissolved in cyclohexane-diethyl ether-methylene chloride (10: 30: 5) hot and the resulting solution cooled to room temperature. In this case, a yellow OeI separates, which is stirred after decanting the solvent for half an hour with 50 ml of absolute diethyl ether. The resulting crystals are filtered off with suction and washed with absolute diethyl ether.

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

One obtains the α, ω-tri-O-acetyl-desoxy-1-propionylamino-D-mannos-3-0-yl-acetyl-L-alanyl-D-isoglutamine-benzhydryl ester as a colorless powder containing 1.11 mol of water contains.

· 1.11 H ₂ O (804.81) Ber. C, 56.71, H, 6.31, N, 6.96, 0, 30, 03, H, _2, O, 2.48; C, 56.94, H, 6.36, N, 7.22, 0, 30, 15, H, _2, O 2.48

on [α] ρ - + 5.5 + 2.7 ° (c - 0.365, methanol),

R- - 0.49 (chloroform: methanol - 9: 1), R- «0.79 (chloroform: methanol -4: 1).

Example 2 ; 0.5 g (0.57 mmol) of 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine-benzhydryl ester (α, B mixture) 1.33 mol of water are dissolved in 20 ml of methanol-tetrahydrofuran (1: 1) with 0.1 g 5proz. Palladium carbon as a catalyst at atmospheric pressure and room temperature hydrogenated. After 1 hour, the catalyst is filtered off and the filtrate is evaporated in vacuo at 30 °. The residue is dissolved in 10 ml of methanol-chloroform (1: 1) and the crude product is precipitated from the resulting solution with diethyl ether. The precipitated crystals are filtered off with suction and washed with diethyl ether.

The product thus obtained is dissolved in 50 ml of bidistilled water, filtered twice through a Millipore filter (Nalgene S, 0.45 and 0.2 μm) and lyophilized. This gives 1,4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine (α, B mixture) as a colorless powder containing 1.92 moles of water;

[a] p - + 19.5 ± 2.0 ° (c - 0.492; water), R - 0.16 (chloroform: methanol: water »70: 30: 5).

The starting material is obtained as follows:

Step 2.1: Analogously to Example 1, 2.11 g (4.07 mmol) of N-propionyl-dem-methylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine containing 1.56 mol of water are obtained, with a total of 2.14 g (11.06 mmol) of diphenyldiazomethane in 42 ml of methanol-dimethoxyethane (1: 1) (18 hours, room temperature) of N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutane. <aminyl-L-alanine benzhydryl ester as a colorless powder containing 2.24 moles of water.

· 2.24 H ₂ O (770.14) Ber. C 54.58 H 6.76 N 9.09 0 29.59 H ₂ O 5.25 Gef. C 54.85 H 6.86 N 9.11 0 29.21 H ₂ O 5.25

20 [ot] * - + 2.9 ± 2 ° (c - 0.478; dimethylformamide),

R. 0.57 (chloroform: methanol - 7: 3),

R_ »0.65 (chloroform: methanol: water 70: 30: 5).

- 46 - ^

Step 2.2: Analogously to Example 1, 1.19 g (1.54 mmol) of N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine benzhydryl ester (α, β mixture) containing 2.24 mol of water are obtained , with 0.7 ml (7.4 mmol) of acetic anhydride in 12 ml of pyridine (20 hours, room temperature) of 1,4,6-tri-O-acetyl-N-propionyl-desmethyl-muramyl-L-alanyl-D-isoglutaminyl L-alanine benzhydryl ester (α, β mixture) as a colorless powder containing 1.33 moles of water.

C4iH53N ₅ O ₁₅ · 1.33 H ₂ O (879.85)

Ber. C 55.97 H 6.40 N 7.96 0 29.69 H ₂ O 2.72 ref. C 56.14 H 6.36 N 7.98 O 29.56 H ₂ O 2.72

[et] * «+ 23.8 + 2.1 ° (c - 0.478; dimethylformamide), R _f » 0.36 (chloroform: methanol 9: 1), R_ 0.69 (chloroforra: methanol · 4: 1) , R.-0.87 (chloroform: methanol - 7: 3).

Example 3 Analogously to Example 2, hydrogenation of 0.5 g (0.64 mmol) of α, o-tri-O-acetyl-N-propionyl-demethylmuramyl-L-alanyl-D-glutamic acid (C) is obtained. -n-butyl ester (C) -benzhydryl ester (mainly a-anomeric) containing 0.73 mol of water in the presence of 0.1 g 5proz. Palladium carbon in 20 ml of methanol-tetrahydrofuran (1: 1) (Ia, B), 4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid (C) -n-butyl ester as colorless crystals from Stnp. 94-96 ° (dec. From chloroform-diethyl ether [1: 6]) containing 0.46 mole of water;

on [<*] £ »+ 45.6 ± 1.8 ° (c - 0.543, methanol),

R. "0.39 (chloroform: methanol" 4: 1), R "0.53 (chloroform: methanol 7: 3), R" 0.59 (chloroform: methanol: water - 70: 30: 5).

The starting material is obtained as follows:

Stage 3.1: Analogously to Example 1, 1.9 g (3.3 mmol) of N-propionyl-dem-methylmuramyl-L-alanyl-D-glutamic acid iC) -n-butyl ester containing 1.03 mol of water are obtained in total 2.5 g (12.85 mmol) of diphenyldiazomethane in 40 ml of methanol-dimethoxyethane (1: 1, 70 hours).

- 47 -

room temperature) N-propionyl-demethylmuramyl-L-alanyl-D-glutamic acid (C) -n-butyl ester (C) -benzhydryl ester as a colorless powder containing 0.99 moles of water.

036Hi ₁₉ N ₃ O ₁₂ x 0.99 H ₂ O (733.62) Ber. C 58.94 H 7.03 N 5.73 0 28.32 H ₂ O 2.43 Vessel C 59.24 H 7.06 N 5.59 O 28.08 H ₂ O 2.43 [o] p - + 12.5 + 1.9 ° (c - 0.522, methanol),

R. «0.46 (chloroform-methanol - 9: 1), R-« 0.63 (chloroform-methanol »4: 1), R-0.88 (chloroform-methanol» 7: 3).

Step 3.2: Analogously to Example 1, 1.15 g (1.57 mmol) of N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid iC) -n-butyl ester (C) -benzhydryl ester, which is still 0, are obtained. Contains 99 mol of water, with 0.70 ml (7.4 mmol) of acetic anhydride in 12 ml of pyridine (16 hours, room temperature) 1.4, ö-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D -glutamic acid (C) -n-butyl ester (C) -benzhydryl ester (α, β-Geraisch) as a colorless powder containing 0.73 mol of water.

CJt ₂ H ₅₅ N ₃ O ₁₅ · 0.73 H ₂ O (855.08) Ber. C 58.99 H 6.60 N 4.91 0 29.50 H ₂ O 1.54 Gef. C 58.96 H 6.72 N 4.76 O 29.49 H ₂ O 1.54 [a] p + 12.6 ± 2.3 ° (c - 0.443; chloroform),

R- "0.72 (chloroform: methanol 9: 1), R-0.96 (chloroform-methanol-4: 1).

Example 4: 4.00 g (5.28 mmol) of N-acetyl- (lo.B), 4,6-tri-O-acetyl-demethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester dissolved in 40 ml of methanol Water (3: 1) are hydrogenated analogously to Example 1 in the presence of 0.8 g of palladium-on-carbon (10%). The catalyst is filtered off, the filtrate is evaporated to dryness and the residue is chromatographed on silica gel (type 60, Merck, 0.063-0.2 mm) (1:50) in the system chloroform-methanol-water (70: 30: 5) (fractions 1-30: 20 ml, from fraction 31: 5 ml). The material contained in fractions 3-45 is collected in 50 ml of double distilled water

4 / dy

dissolved and, after filtration, lyophilized through a Millipore filter (0.2 μ). This gives N-acetyl-1,4,6-tri-O-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine (α, β mixture) as a colorless powder, containing 1.31 mol of water.

C ₂ ^ H37N _If 0i4 »l, 31 H ₂ O (629,17)

Ber. C 45.82 H 6.38 N 8.91 H ₂ O 3.74

C 45.9 H 6.6 N 9.1 H ₂ O 3.7

on [ct] £ - +42 ± Γ (c> 0.768, methanol),

R. 0.65 (n-butanol: pyridine: acetic acid: water 38: 24: 8: 30).

R. 0.56 (ethyl acetate: n-butanol: pyridine: acetic acid: water

" 42: 21: 21: 6: 10), R- * 0.39 (ethyl acetate: acetic acid: water: methanol - 67: 10: 23: 12).

The starting material is obtained as follows: Step 4.1: A solution of 22.8 g (40 mmol) of N-acetyl-demethylmuramyl-L-alanyl-D-isoglutamine in 500 ml of a 1: 1 mixture of 1,2-dimethoxy-ethane 11.6 g (60 mmol) of diphenyldiazomethane are added to methanol and the solution is stirred for 16 hours at room temperature. The red suspension is evaporated at 20 ° and under reduced pressure, and the residue is triturated with diethyl ether until an almost colorless product is obtained. This is dissolved in 100 ml of methanol and brought by the portionwise addition of a 2:! - Mixture of diethyl ether: petroleum ether to crystallize. After several hours of stirring, first at room temperature, then in an ice bath, the crystal mass is filtered off and dried under reduced pressure. This gives the N-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester in the form of cubic crystals. Mp 170 ° (with decomposition),

[o] p - + 14 ± 1 ° (c - 1.5, methanol), R. «0.40 (chloroform: methanol: water 70: 30: 5) and R. 0.66 (ethyl acetate: n-butanol : Pyridine: glacial acetic acid: water - 42: 21: 21: 6: 10).

Stage 4.2: Analogously to Stage 1.2, 3.69 g (5.6 mmol) of N-acetyl

- 49 -

desmethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester, dissolved in 40 ml abs. Pyridine, with 2.04 g (20 mmol) of acetic anhydride (1 hour, room temperature). The clear solution is concentrated at 30 °, the residue taken up in 150 ml of ethyl acetate and the solution extracted twice with 50 ml of water. After drying and evaporation of the solvent remains N-acetyl (la, ß), 4,6-tri-O-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamin benzhydrylester.

[a] £ ° - + 33 ° ± 1 ° (c - 1,159, methanol), R. 0,70 (chloroform: methanol: water - 70: 30: 5), R- «0,48 (n-butanolacetic acid: Water "75: 7.5: 21) and R-0.81 (ethyl acetate: n-butanol: pyridine: acetic acid: water - 42: 21: 21: 6: 10).

Example 5 Analogously to Example 4, 0.60 g (0.76 mmol) of N-acetyl-o-tri-O-acetyl-desmethylmuramyl-L-ce-aminobutyryl-D-isoglutaminebenzhydryl ester, dissolved in 20 ml of dimethoxyethane-water ( 20: 1) in the presence of 0.2 g of palladium on carbon (10%). The residue is purified by chromatography on silica gel in the system chloroform-methanol-water (70: 30: 5). The fractions containing the material are collected, dissolved in 10 ml of dimethoxyethane-water (1: 1), and over a strongly acidic ion exchanger (Dowex® 50, Xe / particle diameter 0.149-0.074 mm, 50 ml) to remove partially formed Na salt. filtered and washed with a total of 150 ml of mixture. The combined filtrates are evaporated on a rotary evaporator at about 30 °, the residue dissolved in 10 ml of a mixture of tert-butanol-water (2: 1) and filtered through a Millipore filter (Fluoropore ·, PTFE, 0.2 μ ) lyophilized. This gives N-acetyl-l ^ .o-tri-O-acetyl-desmethylmuramyl-La-aminobutyryl-D-isoglutamine (α, β mixture) as a colorless powder, containing 1.94 mol of water.

C ₂₅ H ₃ 8NuOi ^ l, 94 mol H ₂ O (653.54) Ber. C 45.95 H 6.49 N 8.57 H ₂ O 5.34 Gef. C 46.2 H 6.6 N 8.3 H ₂ O 5.3

[ct] p - + 34 ± 1 ° (c - 0.511; chloroform),

- 50 -

R-0.15 (chloroform: methanol: water * 70: 30: 5), R-0.58 (ethyl acetate: n-butanol: pyridine: acetic acid: water - 42: 21: 21: 6: 10).

The starting material is obtained as follows:

Stage 5.1: Analogously to Stage 4.2, 0.55 g (0.85 mmol) of N-acetyl-demethylmuramyl-L-ct-aminobutyryl-D-isoglutamine-benzhydryl ester, dissolved in 5 ml of abs. Pyridine, and 0.281 g (2.76 mmol) of acetic anhydride (30 minutes, room temperature) of N-acetyl- (1a, β), 4, o-tri-O-acetyl-demethylmuramyl-L-ct-aminobutyryl-D-iso glutamine benzhydryl ester. R ,. 0.63 (chloroform: methanol: water 70: 30: 5) and R- «0.82 (ethyl acetate: n-butanol: pyridine:

Acetic acid: water »42: 21: 21: 6: 10).

EXAMPLE 6 Analogously to Example 5, 1.02 g (1 mmol) of N-acetyl-1,4,6-tri-O-acetyl-demethylmuramyl-L-aaminobutyryl-D-glutamic acid dibenzehydryl ester are obtained by hydrogenolysis, followed by chromatography Silica gel and treatment with Dowex® 50 (H-form) N-acetyl-1,4,6-tri-O-acetyl-desmethylmuramyl-La-aminobutyryl-D-glutamic acid (o, B mixture) as a colorless, loose powder, Containing 1.38 mol of water.

C25H ₃ 7N ₃ Oi5 »l, 38 H ₂ O (644,44)

Ber. C 46.59 H 6.26 N 6.52 H ₂ O 3.87

Gef. C 46.8 H 5.9 N 6.5 H ₂ O 3.9

[α] ρ - +16 ± 1 ° (ς - 0.666, chloroform), R «0.23 (chloroform: methanol: water 70: 50: 5), R» 0.43 (n-butanol: pyridine: acetic acid : Water - 38: 24: 8: 30), R-0.32 (ethyl acetate: n-butanol: pyridine: acetic acid: Wa8ser - 42: 21: 21: 6: 10).

The starting material is obtained as follows:

Step 6.1: 3.00 g (6.1 mmol) of N-acetyl-demmethylmuramyl-Lo-aminobutyryl-D-glutamic acid, dissolved in 30 ml of a 1: 1 mixture of dimethylformamide and methanol, are treated with excess diphenyldiazomethane at room temperature and with stirring , After 3 days will be

- 51 -

The red suspension was evaporated in a high vacuum at 30 °, the red residue triturated several times with diethyl ether-petroleum ether (1: 3) and the supernatant decanted off. The solid residue is first by chromatography on 600 g of silica gel (type 60, Merck) in the system chloroform-methanol-water (70: 30: 5, inter alia, to remove half ester), then on a second column (100 g) first with ethyl acetate , then ethyl acetate-methanol (3: 1, 5 ml fractions).

The collected material is dissolved in 4.5 ml of methanol and after addition of 70 ml of abs. Dioxane and conventional filtration through a Millipore filter lyophilized. N-acetyl-desmethylmuramyl-L-α-aminobutyryl-D-glutamic acid-dibenzhydryl ester is obtained as a colorless powder containing 0.93 mol of water.

0.93 H ₂ O (842.67) Ber. C 64.14 H 6.34 N 4.99 H ₂ O 1.99 Found C 64.1 H 6.5 N 4.9 H ₂ O 2.0 [a] p ° - + 11.6 ± 2 , 9 ° (c "0.344, methanol), R" 0.55 (ethyl acetate: methanol "4: 1) and R-0.77 (chloroform: isopropanol 1: 1).

Step 6.2: From 2.10 g (2.54 mmol) of N-acetyl-desmethylmuramyl-L-aaminobutyryl-D-glutamic acid dibenzhydryl ester and 1.053 ml (11.43 mmol) of acetic anhydride in 6 ml of absolute pyridine are obtained analogously to Example 1 according to Standing at room temperature for 12 hours the peracetyl compound. Purification is carried out on 450 g of silica gel (type 60, Merck, particle size 0.063-0.200 mm, 5 ml fractions) first with chloroform, then from fraction 20 with chloroform: isopropanol (3: 1). The material contained in fractions 43-106 is collected, dissolved in 5 ml of chloroform, after addition of 40 ml of abs. Dioxan filtered through a Millipore filter (Fluoropore, PTFE, 0.2 μΐη) and then lyophilized. This gives N-acetyl (la, $), 4,6-tri-O-acetyl-desmethylmuramyl-L-α-aminobutyryl-D-glutamic acid dibenzhydrylester as a colorless, loose powder, containing 0.37 mol of water.

- 52 -

C ₅ H H ₅₇ N ₃ O ₁₅ · 0.37 H ₂ O (958.69) Ber. C 63.90 H 6.09 N 4.38 H ₂ O 0.69 Gef. C 63.6 H 6.1 N 4.6 H ₂ O 0.70 [e »] p ° - + 29.6 + 1.7 ° (c - 0.577, methanol), R - 0.69 (n-butanolacetic acid: water 75: 7.5: 21) and R - 0.93 (ethyl acetate: n-butanol: pyridine: acetic acid: water - 42 : 21: 21: 6: 10).

Example 7 Analogously to Example 5, hydrogenolysis of 0.80 g (1 mmol) of N-acetyl-1,4,4-tri-O-acetyl-demethylmuramyl-L-valyl-D-isoglutamine-benzhydryl ester, dissolved in 20% by weight, is obtained ml of dimethoxyethane-water (4: 1), and conventional chromatography of N-acetyl-1,4,6-tri-O-acetyl-demethylmuramyl-L-valyl-D-isoglutamine (α, β-mixture) as a colorless lyophilisate, 0 Containing 9 moles of water.

- +17 ± 1 ° (c - 0.352; chloroform),, 19 (Chloroforra: methanol: water 7 0.28 (n-butanolacetic acid: water 75: 7.5: 21), ssigsäureethylei 42: 21: 21: 6 : 10).

R _f - 0.19 (chloroforra: methanol: water 70: 30: 5),

R- * 0.63 (ethyl acetate: n-butanol: pyridine: acetic acid: water

The starting material is obtained as follows:

Step 7.1: 1.60 g (3.15 mmol) of N-acetyl-demethylmuramyl-L-valyl-D-isoglutamine dissolved in 30 ml of methanol are esterified with excess diphenyldiazomethane (2 hours, room temperature). The product is purified by repeated reprecipitation from methanol-diethyl ether (1: 8). N-acetyl-desmethylmuramyl-L-valyl-D-isoglutamine-benzhydryl ester is obtained.

[a] p - + 13 ° ± 1 ° (c »1.067, methanol), R» 0.56 (chloroform: methanol: water - 70: 30: 5), R_ 0.42 (n-butanolacetic acid: water) 75: 7.5: 21) and R - 0.72 (ethyl acetate: n-butanol: pyridine: acetic acid: water - 42: 21: 21: 6: 10).

- 53 -

Stage 7.2: Analogously to stage 1.2, 0.67 g (1 mmol) of N-acetyldesmethylmuramyl-L-valyl-D-isoglutaminebenzhydryl ester and 0.367 g (3.6 mmol) of acetic anhydride in 5 ml of abs. Pyridine (30 minutes, room temperature) N-acetyl- (1a, β) ↑, δ-tri-O-acetyl-desmethylmuramyl-L-valyl-D-isoglutamine-benzhydryl ester.

[a] p ° «+ 35 ° ± Γ (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).

EXAMPLE 8 Analogous to Example 5, 0.86 g (0.87 mmol) of N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-valyl-D-glutamic acid-dibenzhydryl ester are obtained by hydrogenolysis and customary Chromatography N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-valyl-D-glutamic acid (α, β-mixture) as colorless lyophilisate, containing 1.15 mol of water;

LIS H ₂ O (688,35) Ber. C 48.52 H 6.56 N 6.29 H ₂ O 3.11 Found C 48.9 H 6.5 N 6.7 H ₂ O 3.1 [ct] p ° - + 44 + 1 ° ( c .0.429; water), R "0.08 (chloroform: methanol: water" 70: 30: 5), R "0.48 (n-butanol: pyridine: acetic acid: water - 38: 24: 8: 30), R-0.48 (ethyl acetate: acetic acid: water: methanol - 67: 10: 23: 12).

The starting material is obtained as follows: Step 8.1: 1.80 g (3.34 mmol) of N-acetyl-muramyl-L-valyl-D-glutamic acid, dissolved in 30 ml of methanol, are treated while stirring with excess diphenyldiazomethane. After three hours of stirring at room temperature, the reaction solution is evaporated and the red, oily residue by repeated trituration with diethyl ether-petroleum ether (1: 1) decolorized. The semi-solid material is taken up in 20 ml of dimethoxyethane and precipitated by dropwise addition of a total of 100 ml of petroleum ether. N-acetyl-muramyl-L-valyl-D-glutamic acid dibenzehydryl ester is obtained as a colorless, amorphous powder.

606

- 54 -

I Ct]] T - +33 ± 1 ° (c - 0.489, methanol),

R _f »0.62 (chloroform: methanol: water - 70: 30: 5),

R-0.66 (n-butanolacetic acid: Waseer 75: 7.5: 21).

Step 8.2: 0.845 g (1 mmol) of N-acetyl-muramyl-L-valyl-D-glutamic acid dibenzhydryl ester, dissolved in 10 ml abs. Pyridine are added with stirring at 0 ° with 0.367 g (3.6 mmol) of acetic anhydride and allowed to stand for 45 minutes. Add 1 ml of distilled water and evaporate at 25 ° to dryness. The residue is taken up in ethyl acetate, the solution washed several times with water, dried over sodium sulfate and evaporated. N-acetyl-1,4, o-tri-O-acetyl-muramyl-L-valyl-D-glutamic acid dibenzhydryl ester is obtained as an amorphous powder.

R _f 0.76 (n-butanol: pyridine: acetic acid: water 38: 24: 8: 30), R-> 0.91 (ethyl acetate: n-butanol: pyridine: acetic acid: water - 42:21:21 6:10).

Example 9 : Analogously, the following compounds are obtained: N-acetyl-1,4,6-tri-O-propionyl-muramyl-L-alanyl-D-isoglutaminyl-

L-alanine,

N-acetyl-1,4,6-tri-O-propionyl-muramyl-L-alanyl-D-isoglutaminyl-

L-lysine,

N-acetyl-1,4, o-tri-O-acetyl-muramyl-L-N-methyl-alanyl-D-isoglutamine, N-acetyl-1,4, o-tri-O-acetyl-muramyl-L-a-methyl-alanyl-D-isoglutamine, N-acetyl-1,4,6-tri-0-acetyl-muramyl-L-prolyl-D "-isoglutamin, N-acetyl-1,4, ö-tri-O-acetyl-desmethylmuramyl-L-alanyl-D-r-carboxy-

isoglutamine methyl ester and

1,4, o-tri-O-acetyl-N-benzoyl-desmethylmuramyl-L-a-aminobutyryl-D-isoglutamine.

Example 10 1.45 g (2.53 mmol) of N-benzoyl-demmethylmuramyl-L-alanyl-D-glutamic acid dimethyl ether in 15 ml of pyridine are admixed with 0.1 g of 4-dimethylaminopyridine and 1.5 g (1.24 ml ) Benzoyl chloride. After 20 hours at room temperature, the mixture is heated for 15 minutes at 40 ° and treated after cooling to room temperature with 10 ml of water.

- 55 -

Sm,

Then the solution is evaporated to the syrup in vacuo, taken up with chloroform and extracted successively with 1N hydrochloric acid, 5% NaHCO3 solution and water. After drying (NaaSOi ») and evaporation of the organic phase, a mixture of (la, B), 4,6-tri-O-benzoyl-N-benzoyl-desmethylmuramyl-L-alanyl-D-glutaminsaureditnethylester and (la, ß ), 6-Di-O-benzoyl-N-benzoyl-desmethylmuramyl-L-alanyl-D-glutamic acid.

Chromatography on 150 g of silica gel (Merck, 0.04-0.063 mm) with CHaCla-ethyl acetate (1: 1, fractions 1-4, 500 ml each of eluent) and CH ₂ Cl ₂ -acetic acid ethyl ester (3: 7, fractions 5-8, 500 ml each of the mobile phase), the mixture obtained can be separated. The four compounds obtained have the following R values (each in chloroform: acetone 6: 4): 0.82 (Ia, 4,6-tri-O-benzoyl compound), 0.72 (I, 4 , 6-tri-O-benzoyl compound), 0.55 (Ia, 6-di-O-benzoyl compound and 0.36 (I $, 6-di-O-benzoyl compound).

20 binding). The [aI _n values are in the order named:

+ 40.6 °, -24.8 °, + 33.8 ° and -90.1 ° (each in chloroform). The melting points are in the order named: 92-96 °, 177-178 °, 151-152 ° and 134-136 °.

EXAMPLE 11 Analogously to Example 1, N-acetyl (N-acetyl-1,4, o-tri-O-acetyl-rauramyl-L-valyl-D-isoglutamine-benzhydryl ester (α, β-mixture) is obtained by hydrogenolysis lot, ß), 4,6-tri-O-acetyl-L ^ rauramyl

20 valyl-D-isoglutamine »l, 6 H ₂ O as a colorless powder; [a] tr »56 + 1 °

(c 0.943; water), R _f - 0.4 (dichloromethane! methanol: glacial acetic acid * 20: 5: 1).

The starting material is obtained as follows: Step 11.1: A solution of 2.0 g of N-acetyl-muramyl-L-valyl-D-isoglutamine in 25 ml of methanol and 25 ml of 1,2-dimethoxyethane is treated 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,

- 56 - ·

stirred, filtered off and dried over NaOH pills. The resulting N-acetyl-muramyl-L-valyl-D-isoglutamine-benzhydryl ester melts at 185 ° (dec.), R _f - 0.5 (chloroform: methanol: water - 14: 6: 1), [a] ^ + 38 ° (c 0.912, methanol).

Step 11.2: A solution of 1.2 g of N-acetyl-muramyl-L-valyl-D-isoglutamine-benzhydryl ester in 12 ml of absolute pyridine is added with stirring 0.75 ml of acetic anhydride and allowed to stand at room temperature for 24 hours. The reaction mixture is then poured onto 40 ml of ice-water, whereupon the product crystallizes out. The crystals are filtered off with suction, washed with water and dried. Recrystallization from ethyl acetate and diethyl ether gives N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-valyl-D-isoglutamine-benzhydryl ester, R ,. 0.3 (methylene chloride ^ ethanol 5: 1).

EXAMPLE 12 Analogously to Example 1, N-acetyl (N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-alanyl-D-isoglutamine-benzhydryl ester (ot, $ mixture) is hydrogenolysed by hydrogenolysis. la, ß), 4,6-tri-O-acetyl-muramyl-

L-alanyl-D-isoglutamine »l, 8 ΗσΟ as a colorless lyophilisate;

20 [ci] ^ + 57 + 1 ° (c 0.923; water), R-0.4 (dichloromethane:

Methanol: acetic acid »20: 5: 1). _v The starting material is obtained as follows:

Step 12.1: To 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 added 0.42 g of diphenyldiazomethane and allowed to react for 20 hours stirred at 40 ⁰ C. After evaporation of the solvent, the residue is extracted thoroughly with diethyl ether and water and dried over NaOH pills. N-acetyl-muramyl-L-alanyl-D-isoglutamine-benzhydryl ester is obtained in the form of white crystals. R- "0.6 (methylene chloride: ethanol: water" 14: 6: 1).

Step 12.2: A solution of 2.0 g of N-acetyl-muramyl-L-alanyl-D-isoglutamine-benzhydryl ester in 20 ml of absolute pyridine is added with stirring 1.7 ml of acetic anhydride and for 24 hours at

- 57 -

Room temperature left. The reaction mixture is then poured onto 50 ml of ice-water. The precipitated material is taken up in 100 ml of ethyl acetate, washed with dilute hydrochloric acid and half-saturated Rochsalzlösung and dried over magnesium sulfate. Evaporation of the solvent gives N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-alanyl-D-isoglutamine-benzhydryl ester. R- «0.7 (methylene chloride: methanol 5: 1).

Example 13 ; The following compounds are obtained in an analogous manner: N-benzoylamino-1,4,6-tri-on-hexanoyl-demethylmuramyl-L-alanyl-D-isoglutamine [R-0.67 (chloroform: methanol 17: 3)] , N-propionyl-1,4,6-tri-O-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine, N-acetyl-1,4,6-tri-O-propionyl-muramyl-L-ce-aminobutyryl -D-isoglutamine, N-acetyl-1,4,6-tri-O-acetyl-demethylmuramyl-L-alanyl-D-glutamic acid diamide, 1,4,6-tri-O-acetyl-N-benzoyl-demethylmuramyl -L-alanyl-D-glutamic acid dimethyl ester, N-acetyl-1,4,6-tri-O-acetyl-demethylmuramyl-L-alanyl-D-glutaric acid dimethyl ester, N-acetyl-1,4,6 -tri-O-acetyl-muramyl-La-aminobutyryl-D-isoglutaminyl-L-arginine methyl ether and N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-alanyl-D-isoglutaminyl-N benzyloxycarbonyl-4-thialysine isopropyl ester sulfoxide.

Example 14 : Female MF-2f SPF mice weighing 14-16 g are administered under mild anesthesia with a mixture of equal parts of diethyl ether, ethanol and chloroform with lethal doses (approximately one-one ng plaque forming units [PFU]. ) in the form of 0.05 ml each of a suspension of influenza A / Texas / 1/77 viruses (mouse-adapted strain) intranasally infected.

Each mouse from groups of 10 of each of these mice is given at the time indicated below [days] relative to the day of infection once (single dose) the amount of the respective active substance in 0.05 or in 0.2 ml of a 0.005 wt .% Solution of carboxymethylcellulose sodium salt in double distilled, pyrogen-free water in the case of intranasal or oral administration in the manner mentioned in Table 1.

- 58 -

Jewels of the above-mentioned infected mice are used for the purpose of rontrol, i. they receive a placebo (0.005 wt% solution of carboxymethylcellulose sodium salt).

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

Compound 1-α-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine.

Table 1:

Active substance Application type Application time [days] Percentage of mice still living 23 days after infection, depending on the active substance, amount [mg / kg], statistical significance * P <0.05, ** P £ 0.01 (Vierfel's test), n.t. «Not checked 0.01 0.001 0 = control 0.1 50 * 66 ** 5 I orally +7 n.t. 90 * 80 40 intranasally -7 100 *

- 60 -

Example 15: Non-aqueous single dose for nasal administration Composition:

(Ια, β), 4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid (C) -n-butyl ester 0.03 mg miglyol 812 30.00 mg

Production :

0.03 mg of the active ingredient is added under aseptic conditions in

29.97 mg Miglyol dissolved.

This solution is filled in a commercial disposable nose applicator, which is placed on a fuel can before use.

Example 16: Nasal drops Composition:

(1a, β), 4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid (C) -n-butyl ester thiomersal sodium monohydrogenphosphate * 2H2O sodium dihydrogen phosphate · 12H benzalkonium chloride ethylenediaminetetraacetic acid dinatrium salt (EDTA ) Sodium chloride Demineralized water

Production :

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

0.15 mg mg 0.10 mg mg 0.02 mg - mg 0.30 mg 0.30 mg 10.10 10.10 mg - mg 0.10 mg 0.50 mg 0.50 mg 3.70 mg 4.50 988.30 987.60

- 61 -

In this solution, the drug is then dissolved and supplemented with the remaining entraineralized water.

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

a) glass or plastic containers (a 5 ml or 10 ml) containing a glass or plastic pipette with an elastomeric pipettor,

b) plastic crumpled bottles with a riser and a plastic spraying head,

c) single dose plastic container (content 2-3 drops) or

d) glass or plastic bottles provided with a normalized pumped dosing spray of plastic (without propellant gas).

Example 17 : Nose Ointment

Composition ;

(La, B), 4,6-tri-O-acetyl-N-propionyldesmethylmuramyl-L-alanyl-D-glutamine

acid (C) -n-butyl ester 0.03g

Paraffin oil viscous 20,00 g

white Vaseline 30,00 g

Wool fat, anhydrous 40.00 g

demineralized water 19.97 g

Production :

The fat phase, consisting of paraffin oil, petrolatum and wool fat, is melted together. The aqueous solution of the active ingredient is incorporated at about 50 ^° C. into the fatty phase.

- 62 -

Example 18 : Preparation of 1000 tablets containing 0.5% active ingredient

Composition per 1000 tablets :

(1 ', β), 4,6-tri-O-acetyl-N-propionyl-desmethylmuramyl-L-alanyl-D-glutamic acid

(C) -n-butyl ester 0.5 g

Lactose ground 43.0 g Corn starch 52.0 g Pharmacoat 603® (hydroxypropylmethyl

cellulose containing 28-30% methoxyl groups, supplied by Shinetsu Chemical

Company, Toio, Japan) 3.0 g Aerosil® (colloidal silica,

supplied by Degussa, Frankfurt,

Federal Republic of Germany) 1.0 g Magnesium stearate 0.5 g Production :

The active ingredient and 15 g of lactose are premixed. The premix thus obtained is mixed together with 28 g of lactose and 47 g of corn starch. With the mixture thus obtained and an aqueous solution of the Pharmacoat a granulate-ready mass is prepared, which is granulated, dried and ground. To this end, 5 g of corn starch, aerosil and magnesium stearate are mixed and compressed to give 1000 tablets each weighing 100 mg.

The pellets can be coated enteric-coated in a manner known per se.

Example 19: 0.070 g (0.088 mmol) of N-acetyl-1,3-tri-O-acetyl-muramyl-L, N-methyl-alanyl-D-isoglutamine-benzhydryl ester (ot, B mixture) are obtained analogously to Example 5, but in glacial acetic acid, hydrogenated. The solution is lyophilized after removal of the catalyst. The residue is taken up in 3 ml of chloroform, passed through a Millipore filter (PTFE, 0.2 μιη) and the solution after addition of 60 ml of abs.

547 60

- 63 -

Dioxane lyophilized. This gives N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-N-methyl-alanyl-D-isoglutamine (α, β mixture) as a colorless powder;

[α] £ * - + 34.9 ± l, l ° (c - 0.946, methanol),

R _f 0,2 0.25 (chloroform: methanol: water - 70: 30: 5), R _f 0.43 (acetonitrile: water 3: 1), R. "0.21 (n-butanol." Acetic acid: water 75: 7.5: 21). The starting material is obtained as follows: Step 19.1: 0.50 g (0.99 mmol) of N-acetyl-muramyl-L-N-methyl-alanyl D-isoglutamine (o, B mixture) are prepared analogously to stage 8.1 in N-acetyl

muramyl-L-N-methyl-alanyl-D-isoglutamine-benzhydryl ester;

R_ »0.76 (acetonitrile: water -3: 1), R _f - 0.63 (chloroform: methanol: water - 70: 30: 5).

Step 19.2: The crude product obtained according to step 19.1 is acetylated analogously to step 8.2 and purified by chromatography on silica gel (1: 180) in chloroform-isopropanol (9: 1, 0.8 ml fractions). The uniform fractions are made up of abs. Dioxane lyophilized. This gives N-acetyl-1,4,6-tri-O-acetyl-muramyl-LN-methyl-alanyl-D-isoglutamine-benzhydryl ester (α, β-mixture) as a colorless powder; R _f 0.61 (n-butanol: acetic acid: water 75: 7.5: 21), R "0.20 (chloroform: 18% propanol: ice 70: 8: 2).

Example 20: 0.32 g (0.40 mmol) of N-acetyl-1,4,6-tri-O-acetyl-amino-isobutyryl-D-isoglutamine-benzhydryl ester (α, β-mixture) dissolved in 8 ml Glacial acetic acid, are hydrogenated analogously to Example 5, but in glacial acetic acid. The catalyst is filtered off and the solution is filtered through a Millipore filter (PTFE, 0.2 μm). After lyophilization, N-acetyl-1,4,6-tri-O-acetyl-muramyl-α-amino-isobutyryl-D-isoglutamine (α, β mixture) remains as a colorless powder containing 0.81 mole of water;

[a] p - + 55.8 ± 1.1 ° (c - 0.923, methanol),

R _f 0.40 (chloroform: methanol.'water 70: 30: 5),

- 64 -

R-0.70 (ethyl acetate: pyridine: acetic acid: water

62: 21: 6: 11), R "0.73 (ethyl acetate: n-butanol: pyridine: acetic acid: water - 42: 21: 21: 6: 10).

The starting material is obtained as follows:

Step 20.1: 0.50 g (0.99 mmol) of N-acetyl-muramyl-α-amino-isobutyryl-D-isoglutamine (α, β mixture) are esterified with diphenyldiazomethane analogously to step 8.1. The crude product [R-0.48 (chloroform: methanol: water »70: 30: 5)] is peracetylated analogously to step 8.2. The reaction solution is evaporated to dryness in vacuo, taken up in water and lyophilized. Purify on silica gel (1: 100) in chloroform-methanol (9: 1, 1 mL fractions). The pure fractions are combined, taken up in 5 ml of chloroform, filtered (PTFE, 0.2 μm) and then evaporated to dryness. The residue is in abs. Dioxane taken and lyophilized. N-acetyl-1,4, δ-tri-O-acetyl-muramyl-α-amino-isobutyryl-D-isoglutamine-benzhydryl ester (α, β-mixture) is obtained as a colorless powder containing 0.73 mole of water;

[a] p - + 45.7 ± 1 ° (c - 0.993, methanol), R - 0.80 (chloroform: methanol: water 70: 30: 5), R - »0.32 (chloroform: methanol 9 :1),

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

Example 21: 0.30 g (0.37 mmol) of N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-prolyl-D-isoglutamine-benzhydryl ether (α, β mixture) are prepared analogously to Example 20 hydrogenated in glacial acetic acid. After filtration (PTFE, 0.2 μΐη) and lyophilization, N-acetyl-1,4,6-tri-O-acetylmuramyl-L-prolyl-D-isoglutamine (α, β-mixture) remains as a colorless powder;

[a] p - + 51.4 + 1 ° (c - 0.956, methanol), R_ »0.20 (chloroform: methanol: water 70: 30: 5), R -« 0.39 (acetonitrile: water - 3 :1).

- 65 -

247 60-

The starting material is obtained as follows:

Stage 21.1: 0.50 g (0.934 mmol) of N-acetyl-muramyl-L-prolyl-D-isoglutamine [α, β-mixture] are esterified with diphenyldiazomethane analogously to stage 8.1. After the usual work-up, N-acetyl-muramyl-L-prolyl-D-isoglutamine-benzhydryl ester (α, β-mixture) is obtained as a colorless powder;

[a] £ ° - + 30.4 + 1.6 ° (c - 0.608, methanol),

R _f 0.63 (chloroform: methanol: water 70: 30: 5).

Stage 21.2: N-acetyl-muramyl-L-prolyl-D-isoglutamine-benzhydryl ester is peracetylated analogously to stage 8.2. After lyophilization from abs. Dioxane remains N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-prolyl-D-isoglutamine-benzhydryl ester hydrate (α, β-mixture) as colorless

Powder;

on [a] p - + 32.2 + 1.6 ° (c - 0.645, dimethyl sulfoxide),

R.-0.92 (chloroform: methanol: water 70: 30: 5), R_ - 0.60 (n-butanol: acetic acid: water 75: 7.5: 21).

Example 22: N-acetyl-1,4, o-tri-O-acetyl-desmethylmuramyl-L-alanyl-Dy-benzhydryloxycarbonyl-isoglutamine methyl ester (α, β mixture) are hydrogenated analogously to Example 5, but in glacial acetic acid. After customary work-up, filtration (PTFE, 0.2 μπι) and lyophilization remains N-acetyl-1,4,6-tri-O-acetyl-desmethylmuramyl-L-alanyl-D-7-carboxy-isoglutamin-methyl ester (α, β-mixture) as a colorless powder; ·

R _f 0.66 (chloroform: methanol: water. 'Acetic acid 55: 47: 13: 5), R-0.18 (chloroform: methanol: water 70: 30: 5).

The starting material is obtained as follows:

Step 22.1: 0.216 g (0.40 mmol) of N-acetyl-demethylmuramyl-L-alanyl-D-y-carboxy-isoglutamine (α, β mixture) are esterified analogously to Step 8.1 with diphenyldiazomethane (3 equivalents). The light reddish suspension is filtered after three hours of stirring and the filtrate is evaporated to dryness. The mixture consisting of starting material, mono- and dibenzhydryl ester and decomposition products which are derived from diphenyldiazo

- 66 -

can be separated by chromatography on Rieselgel (1: 100) in the system chloroform-methanol-water (70: 30: 5). After two chromatographs, N-acetyl-demethylmuramyl-L-alanyl-Dy-benzhydryloxycarbonyl-isoglutamine-benzhydryl ester (α, β-mixture) is obtained as a colorless resin having R-values R-0.67 (chloroform: methanol: water 70 : 30: 5) and R- «0.46 (n-butanol: acetic acid: water» 75: 7.5: 21) and N-acetyl-demethylmuramyl-L-alanyl-D-isoglutamine-benzhydryl ester with the R_- value

R _f - 0.13 (chloroform: methanol.'water 70: 30: 5).

Step 22.2: N-Acetyl-deemethylmuraroyl-L-alanyl-D-isoglutamine-benzhydryl ester is esterified in methanolic solution as usual with a solution of diazomethane in diethyl ether. After evaporation, N-acetyl-desmethylmuramyl-L-alanyl-Dy-benzhydryloxycarbonyl-isoglutamin-ethyl ester (α, β-mixture) remains as a colorless resin; R _f 0.56 (chloroform: methanol: water 70: 30: 5).

Step 22.3: Analogously to Section 8.2, N-acetyl-1-methyl-1-methylpyrrolidinyl-N-acetyl-demethylmuramyl-L-alanyl-Dr-benzhydryloxycarbonyl-isoglutamine-methyl ester (α, β-mixture) is obtained. L-alanyl-Dy-benzhydryloxycarbonyl-isoglutamine-methylbenzene (α, β-mixture) as lyophilisate;

R _f 0.67 (chloroform: methanol: water - 70: 30: 5),

R_ 0.35 (n-butanol: Es8acetic acid: water 75: 7.5: 21).

Example 23 Analogous to Example 5, but in glacial acetic acid, one obtains from N-acetyl-1,4, O-tri-O-acetyl-desmethylmuramyl-L-alanyl-Dy-benzhydryloxycarbonyl-isoglutamine-benzhydryl ester (α, β-mixture) N-acetyl-1, 4,6-tri-O-acetyl-desmethylmurarayl-L-alanyl-D-carboxy-isoglutamine as a colorless, loose powder;

on [a] p - + 35.7 ± 1.6 ° (c - 0.613; methanol),

R _f 0.095 (chloroform: methanol: water - 70: 30: 5), R- * 0.093 (n-butanolacetic acid: water 75: 7.5: 21).

- 67 -

The starting material is obtained as follows: Step 23.1: 48.5 mg (0.05 mmol) of N-acetyl-desmethylmuramyl-L-alanyl D-7-benzhydryloxycarbonyl-isoglutamine-benzhydrylester («, ß-mixture)

(see step 22.1) are peracetylated analogously to step 8.2. The

Reaction solution is with little abs. Diluted dimethylformamide and in HV evaporated to dryness. The resulting resin crystallizes Take up in a little methanol (about 10% solution). This gives N-Ace

tyl-1,4, o-tri-O-acetyl-demmethylmuramyl-L-alanyl-D-r-benzhydryloxycarbonyl-isoglutamine-benzhydryl ester (α, B mixture) in the form of colorless

needles; Mp. 198-190 ⁰ C (Zers.), R _f - 0.85 (chloroform: methanol: water - 70: 30: 5), R. »0.23 (chloroform: isopropanol: glacial acetic acid - 70: 8: 2), R_ 0.49 (n-butanol: acetic acid: water 75: 7.5: 21).

Example 24: 0.200 g (0.236 mmol) of 1,4,6-tri-O-acetyl-N-benzoyl-desmethylmuramyl-La-aminobutyryl-D-isoglutamine-benzhydryl ester (α, B mixture) are prepared analogously to Example 5, but in Glacial acetic acid, hydrogenated. After filtration (PTFE, 0.2% w) and lyophilization from abs. Dioxane remains 1,4, δ-tri-O-acetyl-N-benzoyl-desmethylmuramyl-L-O-J-aminobutyryl-D-isoglutamine (α, B mixture) as a colorless powder containing 0.75 mole of water;

on [ö] p - + 33.9 ± 1.1 ° (c - 0.885, chloroform), R »0.44 (n-butanol: acetic acid: water - 75: 7.5: 21), R-« 0.60 (acetonitrile: water - 3: 1).

The starting material is obtained as follows:

Step 24.1: 0.83 g (1.44 mmol) of N-benzoyl-desmethylmuramyl-La-aminobutyryl-D-isoglutamine sodium salt (α, β mixture) are dissolved in the presence of 1.3 mmol of trifluoroacetic acid (90% of theory). as in step 8.1, esterified with diphenyl diazomethane. The crude product is taken up in 20 ml of n-butanol, the solution extracted 3 times with 5 ml of water and the water phases are back-extracted twice with 10 ml of n-butanol. The combined upper phases are dried over Na 2 SO and concentrated to about 5 ml. The product is prepared by adding 150 ml of diethyl

247 60 *

ether precipitated. N-Benzoyl-desmethylmuramyl-L-O-aminobutyryl-D-isoglutamine-benzhydryl ester (α, β-mixture) is obtained as a colorless powder;

[α] ρ ° - + 20.7 + 1 ° (c - 0.483, dimethylformamide), R - * 0.71 (chloroform / methanol / water 70: 30: 5).

Stage 24.2: 0.40 g (0.55 mmol) of N-benzoyl-desmethylmuramyl-L-α-aminobutyryl-D-isoglutamine-benzhydryl ester (<*, 3-mixture) are peracetylated analogously to stage 8.2. The crude product is purified on trickle gel (1: 100) in the system chloroform-isopropanol (9: 1, 0.8 ml fractions). The pure fractions are in abs. Dioxane was added and after filtration (PTFE, 0.2 μιη) lyophilized. This gives 1,4,6-tri-O-acetyl-N-benzoyl-desmethylmuramyl-L-ct-aminobutyryl-D-isoglutaminbenzhydrylester (et, B mixture) as a colorless powder, containing 0.61 mol of water;

on [<*] £ + 40.7 ± 1.1 ° (c - 0.910, methanol), R_ »0.38 (chloroform: methanol: water« 70: 30: 5), R - - 0.90 (acetonitrile .'Water - 3: 1).

Example 25: 1.29 g (1.53 mmol) of N-acetyl-1,3-tri-O-propionyl-muramyl-La-aminobutyryl-D-isoglutamine-benzhydryl ester (α, B mixture) are dissolved in glacial acetic acid and hydrogenated analogously to Example 8. The residue remaining after freeze-drying is taken up in 30 ml of water and extracted twice with 30 ml of diethyl ether each time. The water phase is freed from ether in vacuo, filtered (Nalgene, 0.2 μω) and lyophilized. One obtains N-Acety1-1,4,6-tri-O-propionyl-muramyl-La-aminobutyryl-D-isoglutamine (α, β-mixture) as a colorless, loose powder, containing 1.26 moles of water;

on [ct] p - + 64.5 ± 0.9 ° (c - 1.106, methanol),

R _f 0.53 (chloroform: methanol: water: acetic acid 70: 40: 10: 5), R - 0.46 (n-butanol: acetic acid: water 75: 7.5: 21).

The starting material is obtained as follows:

- 69 -

Step 25.1: 2.03 g (4 mmol) of N-acetyl-muramyl-L-α-aminobutyryl-D-isoglutamine (α, β mixture) are converted into the benzhydryl ester analogously to step 8.1. After the usual workup remains N-acetyl-muramyl-L-ct-aminobutyryl-D-isoglutamine-benzhydryl ester (α, ß-mixture) as a colorless powder;

[α] ρ - + 41.8 ± 1.8 ° (c - 0.548, methanol),

R _f - 0.73 (chloroform: methanol: water »70: 30: 5), R.« 0.78 (acetonitrile.'water · 3: 1).

Step 25.2: 1.70 g (2.53 mmol) of N-acetyl-muramyl-L-α-aminobutyryl-D-isoglutamine-benzhydryl ester (α, B mixture) dissolved in 25 ml abs. Pyridine, are perpropionylated analogously to step 8.2 (90 equivalents of anhydride). After standing for 3 hours at RT, add 30 ml of water and evaporate the whole in HV at 30 ° to dryness. The resinous residue is abs. In 80 ml. Oioxane, filtered (PTFE, 0.2 μm) and lyophilized. N-acetyl-1,4,6-tri-O-propionyl-muramyl-L-ot-aminobutyryl-D-isoglutamine-benzhydryl ester (α, β-mixture) is obtained as a colorless powder containing 0.79 mol of water;

on [α] ρ - + 52.2 + 3.7 ° (c - 0.270, methanol),

R.-0.67 (acetonitrile: water - 3: 1),

R_ - 0.32 (chloroform: isopropanol: acetic acid - 70: 8: 2), R-0.87 (ethyl acetate: n-butanol: pyridine: acetic acid: water - 42: 21: 21: 6: 10).

Example 26: 0.50 g (1 mmol) of N-acetyl-demmethylmuramyl-L-alanyl-D-glutamic acid diamide (α, B mixture) are peracetylated analogously to step 8.2. The crude product obtained after the usual work-up is purified by chromatography on silica gel (1:80) in the system chloroform-methanol-water (70: 30: 5). The uniform fractions are collected, taken up in 50 ml of double distilled water, the solution filtered (Nalgene: 0.2 μm) and lyophilized. This gives N-acetyl-1,4, o-tri-O-acetyl-demmethylmuramyl-L-alanyl-D-glutamic acid diamide (α, β mixture) as a colorless powder, containing 1.69 moles of water;

- 70 - *

p »+ 36.2 ± 1.8 Cc - 0.5; Dimethyl sulfoxide)

R-0.49 (chloroform: methanol: water 70: 30: 5), R-0.20 (n-butanol: acetic acid: water = 75: 7.5: 21).

Example 27: 0.50 g (0.89 mmol) of N-benzoyl-demmethylmuramyl-L-alanyl-D-glutamic acid dimethyl ester (α, β mixture) are peracetylated analogously to step 8.2. The crude product is chromatographed on silica gel (1: 300 3 ml fractions). The eluant used is n-butanol-acetic acid-water (75: 7.5: 21). The pure fractions are collected, dissolved in 10 ml of chloroform-methanol-water (70: 30: 5), filtered (PTFE, 0.2 μπι) and after concentration and addition of 60 ml of abs. Dioxane lyophilized. There remains 1, 4,6-tri-O-acetyl-N-benzoyl-desmethylmuramyl-L-alanyl-D-glutamic acid dimethyl ester (a, (J-Geraisch) in the form of a white powder, containing 1.31 moles of water;

[α] £ ^υ - + 39.2 ± 1 ° (c »0.615, methanol), R-0.68 (n-butanol: pyridine: acetic acid: water *» 62: 21: 6: 11), R- « 0.53 and 0.57 (n-butanolacetic acid: water 75: 7.5: 21).

Example 28: 0.475 g (0.91 mmol) of dimethyl N-acetyl-dem-methylmuramyl-L-ayanyl-D-glutamate (α, β mixture) are peracetylated analogously to step 8.2 and purified by chromatography on silica gel (1: 100; 5 ml fractions) in the system chloroform-isopropanol (70: 8). The pure fractions are collected, in 60 ml abs. Dioxane, filtered (PTFE, 0.2 μm) and lyophilized. This gives N-acetyl-1,4,6-tri-O-acetyl-desmethylmuramyl-L-alanyl-D-glutamic acid dimethyl ester (α, β mixture) as a colorless powder, containing 0.95 mole of water;

[oilcooling + 20.8 ± 2 ° (c "0.491; chloroform), DHO tau

R- »0.55 (chloroform: methanol« 85:15), R »0.37 (n-butanol: acetic acid: water 75: 7.5: 21).

Example 29: To a solution of 0.200 g (0.28 mmol) of N-acetyl-muramyl-La-aminobutyryl-D-isoglutaminyl-L-arginine methyl ester hydrochloride (α, β-mixture) in 2 ml abs. Pyridine is 0.263 ml

-71-.

(2.8 mmol) of acetic anhydride and allowed to stand at RT overnight. The reaction solution is mixed with 15 ml of water and evaporated in vacuo at 30 °. The crude product is purified by chromatography on silica gel (1: 100) in the system chloroform-methanol-water (70: 30: 5). The uniform fractions are collected, taken up in 3 ml of double distilled water, filtered (0.45 μιη) and after addition of 35 ml of abs. Dioxane lyophilized. This gives N-acetyl-l ^ jö-tri-O-acetyl-muramyl-La-aminobutyryl-D-isoglutaminyl-L-arginine methyl ester hydrochloride hydrate (α, β mixture) in the form of a colorless powder, 2 mol Containing water;

R _f - 0.29 (chloroform: methanol: water 70: 30: 5), R-0.60 (ethyl acetate: acetic acid: water: methanol 67: 10: 23: 12).

The starting material is obtained as follows:

Step 29.1: To a solution of 2.35 g (4.64 mmol) of N-acetyl-muramyl-La-aminobutyryl-D-isoglutamine (α, β-mixture) and 1.21 g (4.64 mmol) of L- Arginine methyl ester dihydrochloride in 100 ml abs. To dimethylformamide are successively added 0.84 g (5.6 mmol) of 1-hydroxybenzotriazole monohydrate, 0.51 ml of N-methylmorpholine and finally 1.15 g (5.6 mmol) of dicyclohexylcarbodiimide. After 48 hours of stirring at RT, the yellow suspension is evaporated on a rotary evaporator under high vacuum at 30 ° and the residue is stirred after addition of 200 ml of water. The precipitate is filtered off and the aqueous phase extracted 4 times with 50 ml of water-saturated n-butanol. The aqueous phase is then freed of butanol in vacuo and lyophilized. The residue is purified by chromatography on silica gel (1: 100, 10 ml fractions) in the system chloroform-methanol-water (70: 30: 5). Still present arginine methyl ester hydrochloride is removed by reverse phase chromatography (Opti UPCi ₂ , 40-63 μπι, 1:42) with 0.5 liters of water, the product then with the same volume of acetonitrile-water (9: 1 ) elutes. It is concentrated strongly in vacuo, the solution is sterile filtered (PTFE:

J 61

- 72 -

0.2 μτη) and lyophilized. N-acetyl-muramyl-L-α-aminobutyryl-D-isoglutaminyl-L-arginine methyl ester hydrochloride (ot, (J mixture) is obtained as a colorless powder containing 2.35 mol of water;

on [ce] p - + 18.5 ± 0.8 ° (c - 1.224, methanol), R - «0.29 (ethyl acetate: n-butanol: pyridine: acetic acid: water

- 42: 21: 21: 6: 10), R _f »0.22 (ethyl acetate: ethyl acetate: water: methanol» 67: 10: 23: 12).

Example 30: To 50 mg (0.074 mmol) of N-acetyl-1,4,6-tri-O-propionylmuramyl-La-aminobutyryl-D-isoglutamine (a, f $ mixture, see Example 15) dissolved in 0.5 ml abs. Dimethylformamide, 13.2 mg (0.087 mmol) of 1-hydroxybenzotriazole (containing 11-13% water), 17.6 mg (0.067 mmol) of L-arginine methyl ester dihydrochloride, 8 μΐ (0.071 mmol) of N- Methyl-morpholine and finally 20.8 mg (0.10 mmol) of dicyclohexylcarbodiimide. After two days of stirring at RT, the yellow suspension is evaporated to dryness and the residue is purified twice on silica gel (1: 100) in the system ethyl acetate-n-butanol-pyridine-acetic acid-water (42: 21: 21: 6: 10). This gives N-acetyl-1,4,6-tri-O-propionyl-muramyl-La-aminobutyryl-D-isoglutamine-L-arginine methyl ester hydrochloride (α, B mixture) as a colorless powder; R-0.52 (ethyl acetate: n-butanol: pyridine: acetic acid: water

- 42: 21: 21: 6: 10), R_ »0.40 (chloroform: methanol: water 70: 30: 5).

Example 31: 0.080 g (0.085 mmol) of N ^a - (N-acetyl-1,4,6-tri-O-acetylmuramyl-L-alanyl-D-isoglutaminyl) -N-benzyloxycarbonyl-L-4-thialysine isopropyl ester ( α, β mixture) are dissolved in 8 ml of methanol containing 0.25 ml of 30% hydrogen peroxide solution and allowed to stand for two days at RT. The excess hydrogen peroxide is decomposed by addition of platinum on carbon, the latter is filtered off and the filtrate is evaporated to dryness. The residue is taken up in 90% dioxane, filtered (PTFE, 0.2 μπ>) and

- 73 -

lyophilized. There remains N ^a - (N-acetyl-1,4,6-tri-O-acetyl-muramyl-D-isoglutarainyl) -N-benzyloxycarbonyl-L-4-thia-lysine-isopropyl ester-sulfoxide (α, β-mixture ) as a colorless powder;

R _f * 0.64 (chloroform: methanol: water: eesacetic acid - 70: 40: 10: 5), R "0.75 (acetic acid ether: n-butanol: pyridine: acetic acid: water

- 42: 21: 21: 6: 10).

The starting material is obtained as follows: Step 31.1: To a solution of 0.31 g (0.81 mmol) of N ^e -benzyloxycarbonyl-L-thialysine isopropyl ester hydrochloride and 0.089 ml (: (0.81 mmol) of N-methylmorpholine in 3.5 ml of absolute dimethylformamide are added to 0.82 g (0.97 mmol) of N-acetyl-muramyl-L-alanyl-D-isoglutamine-N-hydroxysuccinimide ester [α, B mixture, ea The residue is combined with 20 ml of ethyl acetate, stirred for one hour at 0.degree., And the insoluble dicyclohexylurea is filtered off with suction.The residue which is left after evaporation is dissolved in 50 ml of water-saturated n.sup. The upper phases are combined, concentrated to about 5 ml, and the whole is lyophilised after the addition of 45 ml of absolute dioxane, leaving N- (N-acetyl-muralhyl) -butanol. L-alanyl-D-isoglutaminyl) -N-benzyloxycarbonyl-L-thialysine-isopropylestef (α, β-Ger aisch) as a colorless powder which still contains some dicyclohexylurea; Rf 0.46 (ethyl acetate: acetic acid: water: methanol

67: 10: 23: 12), R-0.69 (ethyl acetate: n-butanol: pyridine: acetic acid: water

- 42: 21: 21: 6: 10).

Step 31.2: 0.38 g (0.47 mmol) of N ^a - (N-acetyl-muramyl-L-alanyl-D-isoglutaminyl) -N-benzyloxycarbonyl-L-thialysine isopropyl ester (et, B mixture) in a mixture of 0.40 ml abs. Dimethylformamide and 4 ml abs. Pyridine, treated with 0.20 ml (2.1 mmol) Essigsa'ureanhydrid and allowed to stand for 1} days at RT. The reaction solution is evaporated and purified by chromatography

. ». 247 it

Silica gel (1: 200) in the system chloroform-methanol (9: 1). The

unitary fractions are united. N - (N-acetyl, 4,6-tri-O-acetyl-muramyl-L-alanyl-D-isoglutaminyl) -N-benzyloxy-carbonyl-L-thialysine isopropyl ester hydrate (α, β-mixture) as a colorless powder;

R- - 0.90 (chloroform: methanol: water: acetic acid 55: 47: 13: 5), R-0.81 (ethyl acetate: n-butanol: pyridine: acetic acid: water

«42: 21: 21: 6: 10), R _f » 0.39 (n-butanol: acetic acid: water »75: 7.5: 21).

Example 32 Analogously to Example 2, 1.35 g (1.61 mmol) of N-propionyl-1,4,6-tri-O-propionyl-desmethylmuramyl-L-alanyl-D-isoglutamine- (C) -benzhydryl ester are obtained containing 0.68 mol of water by catalytic hydrogenation with 0.2 g of 10% palladium carbon in 50 ml of methanol-tetrahydrofuran (1: 1) after lyophilization of N-propionyl-1,4,6-tri-0-propionyl- desmethylmuramyl-L-alanyl-D-isoglutamine (α, β mixture) as a colorless powder containing 1.55 moles of water; [o] J ° »+ 41.7 ± 2.1 ° (c - 0.484; water),

R "0.20 (chloroform: methanol - 7: 3), R- - 0.31 (chloroform: methanol: water »70: 30: 5). Example 33: Analogously to Example 28, 1,4,6-tri-O-acetyl-N-pro-

pionyl-desmethylmuramyl-L-alanyl-D-glutamic acid diethyl ester (α, β) mixture.

Example 34: 1.45 g (1.76 mmol) of N-acetyl-1,4,4,6-tri-O-propionyl-muramyl-L-alanyl-D-isoglutaminyl-L-alanine benzyl ester (α, β mixture ), dissolved in 100 ml of glacial acetic acid, are hydrogenated as usual in the presence of palladium on carbon (10%), filtered and the solution is lyophilized. The residue is taken up in 50 ml of double distilled water and filtered (Nalgene, 0.45 μπι). After lyophilization, N-acetyl-1,4,6-tri-O-propionyl-muramyl-L-alanyl-D-isoglutaminyl-L-alanine (α, β mixture) remains as a colorless powder containing 1.52 moles of water ;

- 75 - *

⁰ - + 42.2 ± 1.5 * - (c - 0.657, methanol),

R.-0.15 (chloroform-methanol: water 70: 30: 5), R.-0.26 (n-butanol: acetic acid: Wa8ser 75: 7.5: 21), R - 0.42 (Ethyl acetate: n-butanol: pyridine: acetic acid: water - 42: 21: 21: 6: 10).

The starting material is obtained as follows:

Step 34.1: 1.46 g (2.23 mmol) of N-acetyl-muramyl-L-alanyl-D-isoglutaminyl-L-alanine-benzyl ester (o, β-mixture) are dissolved in 25 ml abs. Pyridine and allowed to stand after addition of 1.74 g (13.4 mmol) of propionic anhydride for 36 hours at RT. The solution is evaporated at 30 ° in a high vacuum, the residue taken up in 50 ml of dioxane-water (1: 2), filtered (PTFE, 0.2 μm) and lyophilized. N-acetyl-muramyl-L-alanyl-D-isoglutaminyl-L-alanine benzyl ester (α, B mixture) is obtained as a colorless powder containing 0.73 mole of water;

[a] p - + 38 ± 1,1 ° (c - 0,900; glacial acetic acid),

R.-0.69 (chloroform: methanol: water - 70: 30: 5), R. 0.33 (chloroform: methanol: water: acetic acid 85: 13: 1.5: 0.5).

Example 35: 1.00 g (0.99 mmol) of N ^a - (N-acetyl-tri-O-propionyl-muramyl-L-alanyl-D-iaoglutaminyl) -N-benzyloxycarbonyl-L-lysine benzyl ester (α, β mixture) are hydrogenated analogously to Example 34 in glacial acetic acid. The lyophilizate obtained after the usual work-up is washed with 30 ml of abs. Diethyl ether thoroughly triturated. The precipitate is filtered off, taken up in 50 ml of double distilled water, filtered (Nalgene, 0.45 μιη) and lyophilized. N-acetyl-1,4,6-tri-O-propionyl-muramyl-L-alanyl-D-isoglutaminyl-L-lysine (a, 3-mixture) is obtained as a colorless powder containing 3.1 mol of water;

70 [a] p - + 49.7 ± 1.1 ° (c - 0.903, methanol),

R. 0.33 (ethyl acetate: acetic acid: water: methanol -.- *

67: 10: 23: 12), R _f »0.21 (chloroform: methanol: water: acetic acid 70: 40: 10: 0.5),

- 76-

R_ 0.08 (ethyl acetate: n-butanol: pyridine: acetic acid: water - 42: 21: 21: 6: 10).

The starting material is obtained as follows:

Step 35.1: To a suspension consisting of 8.10 g (20 mmol) of N-benzyloxycarbonyl-L-lysine benzyl ester hydrochloride and 17.30 g (20 mmol) of N-acetyl-rauramyl-L-alanyl-D-isoglutamine 2.02 g (20 mmol) of triethylamine, dissolved in 20 ml, are added dropwise with vigorous stirring within 15 minutes, with vigorous stirring, over a period of 15 minutes, with vigorous stirring, over a period of 15 minutes, with vigorous stirring, over a period of 15 minutes, while the N-hydroxy-succinimide ester (α, β mixture, about 70%, the residue dicyclohexylurea) is dissolved in 180 ml of dimethylformamide Dimethylformamide, a. After lOstündigem stirring at RT, the insoluble "urea" is filtered off, the filtrate was concentrated to about 30 ml and the product precipitated by portionwise addition of a total of 300 ml of water. The precipitate is filtered off with suction, taken up in 50 ml of dimethylformamide after drying and then reprecipitated analogously with 500 ml of ethyl acetate (water-saturated). The crude product (13.4 g) is purified by chromatography on silica gel (1: 100) in the system chloroform-methanol-water (70: 30: 5). The pure fractions yield N - (N-acetyl-muramyl-L-alanyl-D-isoglutaminyl) -N-benzyloxycarbonyl-L-lyene-benzyl ester (α, β-mixture) containing 1.28 moles of water;

^ + 19 ± 1 ° (c - 0.564; glacial acetic acid),

R 0.47 (chloroform: methanol: water 70: 30: 5), R _f 0.40 (n-butanol: acetic acid: water - 75: 7.5: 21), R 0.77 (ethyl acetate: n- Butanol: pyridine: acetic acid: water « 42: 21: 21: 6: 10).

Step 35.2: 2.00 g (2.37 mmol) of N ^a - (N-acetyl-muramyl-L-alanyl-D-isoglutaminyl) -N-benzyloxycarbonyl-L-lysine-benzyl ester (α, β-mixture) are dissolved in 25 ml abs. Pyridine dissolved and perpropionylated analogously to step 34.1. After standing at RT for 30 hours, it is concentrated by evaporation in a HV at 30 ° and the residue is purified by chromatography on silica gel (1: 100) in the system chloroform-methanol-water (70: 30: 5). The uniform fractions are collected. One obtains N ^ö (N-Ace)

247 SQ '

- 77 -

ε acetyl-tri-O-propionyl-muramyl-L-alanyl-D-ieoglutaminyl) -N-benzyloxycarbonyl-L-lysine benzyl ester (a, B mixture) as a colorless powder containing 0.56 moles of water;

p + 28.8 + 1.0 ° (c - 0.983; methanol),

R _f - 0.90 (chloroformate methanol: water 70: 30: 5), R- - 0.11 (chloroform: isopropanol: E8 acetic acid 70: 8: 2).

Claims (20)

invention claim 1. Process for the preparation of compounds of the formula I, CH ₂ OR ⁶
, O ^l o / o V OR I) R ³ -CH (D) R ⁷ OI II ¹⁰ C - N - C - C - NH - CH - CH ₉ - CH - CU I _c I ο * II (O) ¹¹ NH-CH-C (D wherein the hexose part is derived from D-glucose "D-mannose or D-galactose, η is O or 1" and R is lower alkanoy I or benzoyl, R is lower alkyl or phenyl, R, R and R independently of one another are hydrogen or 5 8 7 Lower alkyl, or R with R together trimethylene and R is hydrogen, R and R are independently hydrogen, Niöderalkanoyl or benzoyl, R is hydrogen or unsubstituted or by phenyl, hydroxy, mercapto or lower alkylthio substituted lower alkyl, R and 12
R independently of one another hydroxyl, amino or lower alkoxy, R hydrogen, carboxy or lower alkoxy 3.1
carbonyl and R is hydrogen or unsubstituted or by amino, lower alkanoylamino, hydroxy, guanidino, lower alkanoyloxy, 2-benzyloxycarbonylaminoethyl-sulfinyl, Z-benzyloxycarbonylaminoethyl-sulfonyl, 2-lower alkoxy - 79 - carbonylamino-ethyl-sulfinyl or 2-lower alkoxycarbonylamino-ethyl-sulfonyl mean substituted lower alkyl, with the proviso that in the compounds wherein the Pyranoseteil of D-glucose is derived and at the same time η 0, R ⁵ _# R ⁷ and R ¹⁰ is Hydrogen, R ⁸ for Q JO Methyl, R is amino and R is hydroxy and in which the radicals R, R and R all three have the same meaning, R, R and R of acetyl and butyryl 2 are different when R is phenyl and at the same time Rür hydrogen, and that R ₁ R and R of acetyl 2 3 are different when R and R are both methyl, and salts of such compounds having at least one salt-forming group, characterized in that a) a compound of the formula II, (II) R ³ -CH (D) I ⁷ 0 cT R ¹⁰ ii l l C-N-C-C-NH-CH-CH "- CH - C- ¹¹ U ¹ R « OR ^D R ^ö (D) O I NH-CH-C- in which at least one of the radicals R, R and, for the preparation of compounds of the formula I in which R and R have the abovementioned meanings except for hydrogen, R ^a and R ^{a is} hydrogen, and the rest of these radicals have the meanings of R, the
Have group R -C (sO) -, R and R, and the remaining substituents have the abovementioned meanings, wherein in a compound of formula II existing free functional groups with the exception of the groups to participate in the reaction, if necessary protected protecting groups are protected with a 12 4
reacting R _f or R transmitted acylating agent, and removing any protecting groups, if necessary, or - NEM the introduced acyl group R, R -C (= "O) b) a compound of the formula III,
CH ₂ OR ⁶ LCR ² (III) in which the substituents have the abovementioned meanings, where free OH groups OR ¹ * and OR ^{6 are} protected by readily cleavable protective groups, or a reactive derivative thereof with a compound of the formula IV, R * JO Ψ 8 Y Γ X-CH-C-N-C-β-NH-CH-CH ₂ -CH-C- (L) NH-CH-C-O R ¹² (IV) wherein X is a reactive esterified hydroxy group, and the remaining substituents have the abovementioned meanings, which are protected in a compound of formula IV free functional groups with the exception of X, if necessary protected by readily cleavable protective groups, reacting and deprotecting existing protective groups, if necessary c) a compound of formula V Ow ^UUW> · R ³ -CH (D) y j- \ NH-CH-CH ₂ -CH-C-J (D) ^O / (V) NH-CH-C wherein q, r, s and t are independently 0 or 1 and wherein the substituents have the abovementioned meanings, wherein in a compound of the formula V existing free functional groups with the exception of the group to participate in the reaction, if necessary by light removable protecting groups are protected, or a reactive carboxylic acid derivative thereof with a compound of formula VI, NH-C (D) H-CH ₂ R ¹ »
-CH-C-0 Γ NH-CH-C- R ¹² (VI) wherein u, ν and χ are independently O or 1 and the remaining symbols and substituents have the abovementioned meanings, wherein in a compound of the formula VI existing free functional groups except for the group to participate in the reaction, if necessary by light cleavable protecting groups are protected, where u, ν and χ are 1, if in the reactant of the formula V q and t are 0, or u are 0 and ν and χ is 1, if q is 1 and t is 0, or u and ν are 0 and χ is 1, when q, r and t are 1 and s is 0 or (for the preparation of compounds of formula I, wherein η is 1 - 33- 247 406 stands) u and χ are O when q, r, s and t are 1, or reacting with a reactive derivative thereof and releasing protective groups present if necessary, or d) for preparing a compound of the formula I in which R ^{9 has} one of the abovementioned meanings other than hydroxyl, and the other substituents have the abovementioned meanings, a compound of the formula VII H ₂ OR ⁶ (VII) R ³ -C Ψ ft 'C-N-C-CN öfc R ^{8 Rl3 Rl} ° H-CH-CH ₂ -CH-C- (D) 8 ^RI1 NH-CH-C R ¹² (L) wherein R ^{13 is} carboxy, and wherein the other substituents have the abovementioned meanings, wherein in a compound of formula VII present free functional groups except R ¹³ are protected if necessary by readily cleavable protecting groups, or a reactive carboxylic acid derivative thereof amidated or esterified and if necessary, splits off existing protective groups, or e) in a compound of the formula I in which at least one of the radicals OR *, OR ⁶ , R ⁸ , C ("O) -R ⁹ , R ¹⁰ , R ¹¹ and Ci-O) -R ¹² is present in a protected form which does not correspond to the definition of the desired end-product which splits off the corresponding protective group (s), 247 · * and, if desired, after carrying out any of the processes ae), converting a compound of formula I obtained into at least one salt-forming group or converting a salt of a compound of formula I into the free compound, and / or a mixture of isomers obtained separates. 2. The method according to item 1, characterized in that one selects the starting materials so that a compound of formula I, wherein the hexose part of D-glucose or D-mannose derived, η is O, R, R and R independently of one another C ", - alkanoyl or 2 3 5 7 Benzoyl, RC ₁ -alkyl or phenyl, R, R and R 5 depend on each other hydrogen or methyl, or R 8V 8 with R together trimethylene and R hydrogen, R Hydrogen, C 1-4 -alkyl or by phenyl, hydroxy, Mercapto or methylthio substituted C ₄ . ^ Alkyl, 9 12 * ^- ~ R and R are independently hydroxy, amino or Lower alkoxy, and R is hydrogen, carboxy or lower alkoxycarbonyl, or a salt of such a compound having at least one salt-forming group except for N-Benzoy1-1,4,6-tri-O-butyry1-demethylmuramyl-L-alanyl-D-isoglutamine , N-benzoyl-1,4,6-t ri-O-acety1-desmethylmuramyl-L-alanyl-D-isoglutamine and N-acetyl-l ^^ - tri-Q-acetyl-muramyl-L-alanyl-D- isoglutamine and its salts. 3. Process according to item 1, characterized in that the starting materials are chosen such that a compound of the formula I in which the hexose part of D-glucose 14 6, η stands for!., R, R and R are independent of MS " 2 each C ₀ "alkanoyl or benzoyl, RC ₄ -alkyl or 3 5 7 * "** Phenyl, R, R and R are independently hydrogen 5 8
or methyl, or R with R together trimethylene and 7 8 R is hydrogen, R is hydrogen, C 1-4 -alkyl or by Phenyl, hydroxy, mercapto or methylthio substituted 9 12
C ₁ "-alkyl, R and R independently of one another hydroxyl, IO
Amino or lower alkoxy, R is hydrogen, carboxy or lower alkoxycarbonyl and R is unsubstituted or substituted by amino, guanidines, lower alkanoyloxy, 2-benzyloxycarbonylamino-ethyl-sulfinyl, 2-benzyloxycarbonylaminoethyl-sulfonyl, 2-lower alkoxycarbonylamino-ethylsulfinyl or 2-lower alkoxycarbonylamino-ethyl-sulfonyl-substituted lower alkyl , or a salt of such a compound with at least one salt-forming group produces. 4. The method according to item 1, characterized in that one selects the starting materials so that a compound of formula I, wherein the hexose part of D-Glucoseableitet, η is 0 or 1 and R ¹ C ₀ , .- alkanoyl 2 3 or benzoyl, R is C- _4- alkyl or phenyl, R is hydrogen or C ₁ -alkyl, R is hydrogen, C ₀ , -alkanoyl or 5 A-o - Benzoyl, R is hydrogen, methyl or together with R Trimethylene, R C ₀ , alkanoyl or benzoyl, R water 8th
or methyl, RC ₁ -alkyl or together with 5 9 R is trimethylene, R is hydroxy, lower alkoxy or amino, R is hydrogen or carboxy, R is unsubstituted or substituted by amino, guanidines or 2-benzyloxycarbonylamino-ethyl-sulfinyl C ₁ alkyl and 12
R is hydroxy, lower alkoxy or amino, or a Salt of such a compound with at least one salt with the exception of N-benzoyl-l, 4,6-tri-O-butyryl-demethylmuramyl-L-alanyl-D-isoglutamine, N-benzoyl-1-tri-O-acetyl-desmethylmuramyl-L- alanyl-O-isoglutamine and N-acetyl-1,4-tri-O-acetyl-muramyl-L-alanyl-O-isoglutamine and their salts. Process according to item 1, characterized in that the starting materials are chosen such that a compound of the formula I in which the hexose portion is derived from D-glucose, η is 0 or 1, RC ₀ --alkanoyl or 2 * ~ ^a 3 Benzoyl, RC " _O- alkyl or phenyl, R is hydrogen or 4
Methyl, R is hydrogen, C _{0 is} - alkanoyl or benzoyl, R is hydrogen, RC ₀ - alkanoyl or benzoyl, R Was- 8 9 hydrogen, R is methyl, ethyl or 2-propyl, R is hydroxy, Amino or C ₁ -alkoxy, R is hydrogen, R is methyl 12
or 4-amino-n-butyl and R is hydroxy or amino _r C ₄ alkoxy, or a salt of such a compound having at least one salt-forming group, with the exception of N-benzoyl-l, 4,6-tri-O-butyryl -desmethylmuramyl-L-alanyl-D-isoglutamine, N-benzoyl-1,4,6-tri-O-acetyl-desmethylmuramyl-L-alanyl-D-isoglutamine and N-acetyl-1,4,6-tri-O-acetyl -muramyl-L-alanyl-D-isoglutamine and their salts. Process according to one of the items 1 to 5, characterized in that the starting materials are chosen such that a compound of the formula I in which at least one of the radicals R and R is lower alkoxy and the other of the radicals Q Λ Ο R and R represent hydroxy, amino or lower alkoxy, or a salt of such a compound with at least one salt-forming group. 7. The method according to any one of items 1 to 6, characterized in that one selects the starting materials such that one obtains a compound of formula I »wherein η is 1 andR is different from hydrogen, or a salt of such a compound with at least one salt-forming Group produces. 8. Method according to one of the points. 1 to 7, characterized in that one selects the starting materials so that a compound of the formula I in which R is C _{0 A} - 5
Alkyl or, together with R, is trimethylene, or a salt of such a compound with at least one salt-forming group produces. 9. The method according to item 1, characterized in that one selects the starting materials so that (IjC, ß), 4,6t tri-O-acetyl-N-propionyl-desmethylrauramyl-L-alanyl-D-glutamic acid (Q ^) - n-butyl ester or a pharmaceutically acceptable salt thereof. ^v 10. The method according to item 1, characterized in that one selects the starting materials such that N-acetyl-1,4,6-tri-O-propionyl-murarayl-L-alanyl-O-isoglutaminyl-L-alanine or a pharmaceutically acceptable salt thereof. ', 11. The method according to item 1, characterized in that one selects the starting materials such that N-acetyl-1,4,6-tri-O-propionyl-muramyl-L-alanyl-D-isoglutaminyl-L-lysine or a pharmaceutically acceptable salt thereof. 12. The method according to item 1, characterized in that one selects the starting materials so that one prepares 1,4,6-tri-Q-benzoy1-N-benzoy1-desmethylmuramy1-L-alany1-D-glutamic acid dimethyl ester. 13. The method according to item 1, characterized in that one selects the starting materials so that one prepares dimethyl 1,6-di-O-benzoy1-N-benzoy1-demmethylmu ramy1-L-alany1-0-glutarainsäure. 14. The method according to item 1, characterized in that one selects the starting materials so that one produces 1,4,6-tri-O-acety1-N-benzoy1-demmethylmuraray1-L-alany1-D-glutamic acid dimethyl ester. 15. Method / la point 1, characterized in that one selects the starting materials so that one produces N-acetyl-1,4,6-tri-0-acety1-demmethylmuramy1-L-alanyl-D-glutamic acid dimethyl ester. 16. The method according to item 1, characterized in that the starting materials are selected such that N-acetyl-1,4,6-tri-O-acetyl-muramyl-LN-methyl-alanyl-D-isoglutamine or a pharmaceutically acceptable Salt of it produces. 17. The method according to item 1, characterized in that one selects the starting materials such that N-acetyl-1,4,6-tri-O-acetyl-muramyl-L-ai-methyl-alanyl-O-isoglutaroin or a pharmaceutically acceptable salt thereof. -89- 18. The method according to item 1, characterized in that one selects the starting materials so that N-acetyl ^ .o-tri-O-acetyl-desmethylmuramyl-L-alanyl-D- ρ - carboxy-isoglutamin-methyl ester or a pharmaceutically usable salt thereof. 19. The method according to item 1, characterized in that the starting materials are selected such that N-acetyl, 4,6-tri-O-acetyl-muramyl-L-ö-arainobutyryl-D-isoglutaminyl-L-arginine methyl ester or a pharmaceutic - table usable salt thereof manufactures. 20. The method according to item 1, characterized in that the starting materials are selected such that N-acetyl-1,4,6-tert-ri-O-acetyl-muramyl-L-alanyl-D-isoglutarainyl-N-benzyloxycarbonyl- 4-thia-lysine isopropyl ester sulfoxide or a pharmaceutically acceptable salt thereof. 21. Method according to item 1, characterized in that the starting materials are selected such that N-acetyl-, δ-tri-O-propionyl-muramyl-L-o-C-aminobutyryl-D-isoglutamine-L-arginine methyl ester or a pharmaceutically acceptable acid addition salt thereof.