EP0000330B1 - Lipopeptides, process for their preparation and pharmaceutical compositions containing them - Google Patents

Description

worin R, und R₂ je einen gesättigten oder ungesättigten, aliphatischen oder gemischt aliphatischcycloaliphatischen, gegebenenfalls auch durch Sauerstoffunktionen substituierten, Kohlenwasserstoffrest mit 11-21 C-Atomen, R₃ Wasserstoff oder den Rest R₁―CO―O―CH₂― wo R, die gleiche Bedeutung hat, und X eine peptidisch gebundene natürliche aliphatische Aminosäure mit freier, veresterter oder amidierter Carboxylgruppe, oder eine Aminosäurensequenz von 2-10 natürlichen aliphatischen Aminosäuren, deren terminale Carboxylgruppe in freier, veresterter oder amidierter Form vorliegt, bedeuten, wobei die mit ^* bzw. ^** bezeichneten Asymmetrie-Zentren die absolute R- bzw. S- oder R-Konfiguration besitzen, und gegebenenfalls Gemische der an den C^**-Atomen epimeren R- und S-Verbindungen, und gegebenenfalls Salze und Komplexe dieser Verbindungen, sowie Verfahren zu ihrer synthetischen Herstellung, und pharmazeutische Präparate enthaltend einen oder mehrere dieser Lipopeptide zusammen mit einem pharmazeutischen Trägermaterial und gegebenenfalls zusammen mit anderen pharmazeutischen Verbindungen.The invention relates to new lipopeptides of the formula

wherein R, and R ₂ each represent a saturated or unsaturated, aliphatic or mixed aliphatic-cycloaliphatic, optionally also substituted by oxygen functions, hydrocarbon radical having 11-21 carbon atoms, R ₃ hydrogen or the radical R ₁ ―CO ― O ― CH ₂ - where R , has the same meaning, and X is a peptide-bonded natural aliphatic amino acid with free, esterified or amidated carboxyl group, or an amino acid sequence of 2-10 natural aliphatic amino acids whose terminal carboxyl group is in free, esterified or amidated form, the with ^* or ^** designated asymmetry centers which have the absolute R or S or R configuration, and optionally mixtures of the R and S compounds which are epimeric on the C ^** atoms, and optionally salts and complexes of these compounds, and methods for their synthetic production, and pharmaceutical preparations containing one or more of these lipopeptides together with one pharmaceutical carrier material and optionally together with other pharmaceutical compounds.

oder entsprechende Formeln mit verkürzter Polypeptid-Kette zukommen dürfte, wobei Y" Y₂, Y₃ Acylreste verschiedener höherer gesättigter und ungesättigter Fettsäuren, wie etwa solcher mit 14-19 C-Atomen und anderer, darstellen. Dieselbe Struktur liegt auch dem Murein-Protein als solchem zugrunde (loc.cit.), wobei die absolute Konfiguration an den beiden asymmetrischen Kohlenstoffatomen der obigen Formel nicht untersucht wurde. Sowohl das Murein-Lipoprotein wie seine genannten Abbauprodukte zeigen in vitro eine mitogene Wirkung gegenüber Mäuse-Lymphocyten. [Vgl. auch Z. Immun.-Forschung, Vol. 153, pp. 11-22 (1977)].Lipopeptides have already been described as degradation fragments of lipoproteins. For example, Hantke and Braun (Eur. J. Biochem. 34, 284-296 (1973) were able to isolate lipopeptides or lipopeptide mixtures in an impure form from the murein lipoprotein of the outer cell wall of Escherichia coli by enzymatic degradation following structure

or corresponding formulas with a shortened polypeptide chain, where Y "Y ₂ , Y _{3 represent} acyl residues of various higher saturated and unsaturated fatty acids, such as those with 14-19 C atoms and others. The same structure also lies in the murein protein as such (loc.cit.), the absolute configuration of the two asymmetric carbon atoms of the above formula not being investigated, and both the murein lipoprotein and its degradation products show a mitogenic effect on mouse lymphocytes in vitro. [See also Z. Immun. Research, vol. 153, pp. 11-22 (1977)].

zugrunde. Als Produkte einer enzymatischen Spaltung von Naturprodukten waren diese Fragmente keine in ihrer Zusammensetzung wohl definierten Produkte und stellten offensichtlich komplizierte Gemische von Kondensationsprodukten aus dem genannten Peptidanteil und sehr verschiedenen Acylderivaten des Glyzerylcysteins dar.As can be seen, the degradation lipopeptides obtained from murein protein contain the "glycerylcysteine"

underlying. As products of an enzymatic cleavage of natural products, these fragments were not products with a well-defined composition and were obviously complicated mixtures of condensation products from the peptide portion mentioned and very different acyl derivatives of glycerylcysteine.

The lipopeptides of the present application now differ from the named degradation products of murein proteins in several respects: they represent lipopeptides of precisely defined, uniform chemical constitution and configuration, which are synthetically accessible and are therefore suitable for therapeutic use; the group they form includes compounds in which the rest of the erythritol takes the place of the glycerol portion in the "glycerylcysteine" and / or those in which an amino acid sequence other than that given above for the known degradation-lipopeptide mixtures in formula (11), or in its place there is also only a single amino acid, and finally also derivatives such as amides and esters of the terminal carboxyl group.

In the acyl radicals R, CO, R ₂ CO in the compounds of the formula and their derivatives, R _i and R _{2 are} saturated or unsaturated, aliphatic or aliphatic-cycloaliphatic, optionally also substituted by oxygen functions, hydrocarbon radicals with 11-21 carbon atoms, ie the acyl radicals are derived from saturated or unsaturated, aliphatic or cycloaliphatic, optionally oxygenated in the hydrocarbon radical with 12-22, preferably with 14-18, carbon atoms. The saturated or unsaturated fatty acids, such as lauric acid, myristic acid, palmitic acid, margaric acid, stearic acid, arachic acid, oleic acid, elaidic acid, linoleic acid, a- and β-elaostearic acid, stearolic acid, a-linolenic acid, and also among the cycloaliphatic-aliphatic, for example, are to be mentioned as such the dihydrosterculic acid, malvalic acid, hydnocarpusic acid, and chaulmoograsic acid. Oxygenated acids of this type, which are also suitable for the acyl radicals R, CO and R ₂ CO, are, for example, those by epoxidation of the above-mentioned olefinic fatty acids and. Resulting cycloaliphatic-aliphatic acids, for example the δ, i-epoxystearic acid, furthermore derivatives of the abovementioned acids which have, for example, one or more hydroxyl groups, such as, for example, ricinoleic acid.

In the compounds of the formula (I) or their diastereomer mixtures and their salts or complexes, the groups R ₁ and R ₂ can be identical to one another or different from one another. Preference is given to compounds in which all three or four acyl radicals are identical, and in particular those in which these represent the palmitoyl, stearoyl or oleoyl radical.

The peptide sequence X in the compounds of the formula (I) or in their salts is composed of a maximum of 10 natural aliphatic amino acids, preferably at least half carrying a hydrophilic group, such as, in particular, hydroxyl, amino, carboxyl, carbamide, guanidino or imidazolyl groups. Of the ionic amino acids in this category, aspartic and glutamic and oxyglutamic acids bearing the acidic groups are to be emphasized, and lysine, ornithine, arginine and histidine are among the basic groups bearing. Amides with a neutral character are especially the amides, such as asparagine, glutamine and hydroxyl groups, primarily serine and threonine.

If X in formula (I) represents the amino acid mentioned, it is also e.g. bearing one of said hydrophilic groups, e.g. especially serine or threonine.

Among the amino acids that can go into the above sequence and that do not carry any hydrophilic groups, the unsubstituted ones should be mentioned in particular, e.g. Glycine, alanine, valine, norvaline, leucine, isoleucine, but also some substituted ones that have a non-hydrophilic character, such as methionine.

The order of the amino acids mentioned in the peptide chain X can be arbitrary, but preference is given to those sequences in which all amino acids with hydrophilic groups are bonded directly to one another, and in turn those in which those sequences of the hydrophilic amino acids are attached to the carboxyl group of the cysteine in Triacyl-Glycerylcystein or Tetraacyl-Erythritylcystein part is bound.

worin R,, R₂ die gleichen Bedeutungen besitzen und X die für Formel (I) gegebene Bedeutung hat, aber im Falle der Aminosäurensequenz 2-5 Aminosäuren aufweist, in welcher vorzugsweise mindestens die Hälfte eine hydrophile Gruppe tragen, sowie ihre Salze und Komplexe. Unter diesen Lipopeptiden seien jene hervorgehoben, deren Aminosäurensequenz in der Gruppe X derjenigen der obengenannten durch Abbau von Murein-Lipoprotein erhaltenen Lipopeptide d.h. solcher der Formel (II), oder einer sich davon ableitenden verkürzten Kette entspricht. Es seien insbesondere folgende Lipopeptid-Typen erwähnt.

• (AcGCT=Acyl-Glyzerylcystein-Teil gemäss Formel IV):
• AcGCT-Ser-Ser-Asn-Ala-Lys-OH
• AcGCT-Ser-Ser-Asn-Ala-OH
• AcGCT-Ser-Ser-Asn-OH
• AcGCT-Ser-Ser-OH,

sodann die entsprechenden Typen, in welchen Threonin, Glutamin oder Asparagin das Serin ersetzen; ferner die Verbindungstypen der Art:

• AcGCT-Ser-Ser-Asn-Ala-G lu-OH
• AcGCT-Ser-Ser-Phe-Ala-Glu-OH
• AcGCT-Ser-Ser-Phe-Ala-OH
• AcGCT-Ser-Ser-Phe-OH

und die entsprechenden mit Threonin, Glutamin oder Asparagin als Austauschaminosäuren für Serin, und die Amide und Carbonsäureester mit terminaler Carbamid- bzw. Estergruppe, wobei in erster Linie die Verbindungen in Betracht zu ziehen sind, in denen die Acylreste R,-CO, R₂―CO im Acyl-Glyzerylcystein-Teil gleich sind und Palmitoyl, Stearoyl oder Oleoyl darstellen, und alle diejenigen, in denen R,-CO und R₂―CO verschieden sind und z.B. Palmitoyl, Stearoyl oder Oleoyl bedeuten, und diese Reste in beliebiger Kombination und/oder Variation vorkommen, wie die in der folgenden Tabelle angeführten, wobei der Glyzerylcystein-teil entsprechend den N- und O-Acylsubstituenten als N-Acyl-0-0-di-Acyl-Cys abgekürzt geschrieben ist:

• N-Palmitoyl-O.O-di-Palmitoyl-Cys-Ser-Ser-Asn-Ala-Lys-OH
• N-Palmitoyl-0.0-di-Palmitoyl-Cys-Ser-Ser-Asn-Ala-OH
• N-Palmitoyl-0.0-di-Palmitoyl-Cys-Ser-Ser-Asn-OH
• N-Palmitoyl-0.0-di-Palmitoyl-Cys-Ser-Ser-OH
• N-Palmitoyl-0.0-di-Palmitoyl-Cys-Ser-OH
• N-Stearoyl-O.O-di-Stearoyl-Cys-Ser-Ser-Asn-Ala-Lys-OH
• N-Stearoyl-O.O-di-Stearoyl-Cys-Ser-Ser-Asn-Ala-OH
• N-Stearoyl-O.O-di-Stearoyl-Cys-Ser-Ser-Asn-OH
• N-Stearoyl-0.0-di-Stearoyl-Cys-Ser-Ser-OH
• N-Stearoyl-O.O-di-Stearoyl-Cys-Ser-OH
• N-Oleoyl-O.O-di-Oleoyl-Cys-Ser-Ser-Asn-Ala-Lys-OH
• N-Oleoyl-O.O-di-Oleoyl-Cys-Ser-Ser-Asn-Ala-OH
• N-Oleoyl-O.O-di-Oleoyl-Cys-Ser-Ser-Asn-OH
• N-Oleoyl-0.0-di-Oleoyl-Cys-Ser-Ser-OH
• N-Oleoyl-O.O-di-Oleoyl-Cys-Ser-OH
• N-Lauroyl-0.0-di-Lauroyl-Cys-Ser-Ser-Asn-Ala-Lys-OH
• N-Lauroyl-0.0-di-Lauroyl-Cys-Ser-Ser-Asn-Ala-OH
• N-Lauroyl-O.O-di-Lauroyl-Cys-Ser-Ser-Asn-OH
• N- Lauroyl-O.O-di-Lauroyl-Cys-Ser-Ser-OH
• N-Lauroyl-O.O-di-Lauroyl-Cys-Ser-OH

ferner die Verbindungen, die sich aus der Substitution des Serins durch Threonin, Glutaminsäure, Glutamin, Asparaginsäure oder Asparagin, aus der Substitution des Lysins durch Glutaminsäure, und solche, die sich aus der gleichzeitigen Substitution von Lysin durch Glutaminsäure und Asparagin durch Phenylalanin ergeben.An important group [group I] of lipopeptides according to the present invention are those in which the peptide chain X consists of a maximum of 5 amino acids. Among these are in particular those of the formula

wherein R ,, R _{2 have} the same meanings and X has the meaning given for formula (I), but in the case of the amino acid sequence has 2-5 amino acids, in which at least half preferably carry a hydrophilic group, and their salts and complexes. Among these lipopeptides, those are to be emphasized whose amino acid sequence in group X corresponds to that of the above-mentioned lipopeptides obtained by degradation of murein lipoprotein, ie those of the formula (II), or a shortened chain derived therefrom. The following types of lipopeptides should be mentioned in particular.

• (AcGCT = acyl-glycerylcysteine part according to formula IV):
• AcGCT-Ser-Ser-Asn-Ala-Lys-OH
• AcGCT-Ser-Ser-Asn-Ala-OH
• AcGCT-Ser-Ser-Asn-OH
• AcGCT-Ser-Ser-OH,

then the corresponding types in which threonine, glutamine or asparagine replace the serine; also the connection types of the type:

• AcGCT-Ser-Ser-Asn-Ala-Glu-OH
• AcGCT-Ser-Ser-Phe-Ala-Glu-OH
• AcGCT-Ser-Ser-Phe-Ala-OH
• AcGCT-Ser-Ser-Phe-OH

and the corresponding ones with threonine, glutamine or asparagine as exchange amino acids for serine, and the amides and carboxylic acid esters with a terminal carbamide or ester group, whereby primarily the compounds in which the acyl radicals R, -CO, R ₂ ―CO in the acyl-glycerylcysteine part are the same and represent palmitoyl, stearoyl or oleoyl, and all those in which R, -CO and R ₂ ―CO are different and mean, for example, palmitoyl, stearoyl or oleoyl, and these radicals in any combination and / or variation, such as those listed in the following table, the glycerylcysteine part being abbreviated as N-acyl-0-0-di-acyl-Cys corresponding to the N- and O-acyl substituents:

• N-palmitoyl-OO-di-palmitoyl-Cys-Ser-Ser-As-Ala-Lys-OH
• N-palmitoyl-0.0-di-palmitoyl-Cys-Ser-Ser-Asn-Ala-OH
• N-palmitoyl-0.0-di-palmitoyl-Cys-Ser-Ser-Asn-OH
• N-palmitoyl-0.0-di-palmitoyl-Cys-Ser-Ser-OH
• N-palmitoyl-0.0-di-palmitoyl-Cys-Ser-OH
• N-stearoyl-OO-di-stearoyl-Cys-Ser-Ser-As-Ala-Lys-OH
• N-stearoyl-OO-di-stearoyl-Cys-Ser-Ser-Asn-Ala-OH
• N-stearoyl-OO-di-stearoyl-Cys-Ser-Ser-Asn-OH
• N-stearoyl-0.0-di-stearoyl-Cys-Ser-Ser-OH
• N-stearoyl-OO-di-stearoyl-Cys-Ser-OH
• N-oleoyl-OO-di-oleoyl-Cys-Ser-Ser-Asn-Ala-Lys-OH
• N-oleoyl-OO-di-oleoyl-Cys-Ser-Ser-Asn-Ala-OH
• N-oleoyl-OO-di-oleoyl-Cys-Ser-Ser-Asn-OH
• N-oleoyl-0.0-di-oleoyl-Cys-Ser-Ser-OH
• N-oleoyl-OO-di-oleoyl-Cys-Ser-OH
• N-Lauroyl-0.0-di-Lauroyl-Cys-Ser-Ser-Asn-Ala-Lys-OH
• N-lauroyl-0.0-di-lauroyl-Cys-Ser-Ser-Asn-Ala-OH
• N-Lauroyl-OO-di-Lauroyl-Cys-Ser-Ser-Asn-OH
• N-Lauroyl-OO-di-Lauroyl-Cys-Ser-Ser-OH
• N-lauroyl-OO-di-lauroyl-Cys-Ser-OH

furthermore the compounds which result from the substitution of the serine by threonine, glutamic acid, glutamine, aspartic acid or asparagine, from the substitution of the lysine by glutamic acid, and those which result from the simultaneous substitution of lysine by glutamic acid and asparagine by phenylalanine.

• N-Myristoyl-O.O-di-Palmitoyl-Cys-Phe-Phe-Asn-Ala-Lys-OH
• N-Lauroyl-O.O-di-Palmotoyl-Cys-Ser-Ser-Asn-Ala-Glu-OH
• N-Stearoyl-O.O-di-Palmitoyl-Cys-Ser-Ser-Asn-Ala-Ala-OH

und die sich durch Ersatz von Ser durch Phe oder von Phe durch Ser ergebenden Verbindungen.Among the compounds with different acyl residues on N and O atoms in the glycerylcysteine residue, the following should be emphasized:

• N-myristoyl-OO-di-palmitoyl-Cys-Phe-Phe-Asn-Ala-Lys-OH
• N-lauroyl-OO-di-palmotoyl-Cys-Ser-Ser-Asn-Ala-Glu-OH
• N-stearoyl-OO-di-palmitoyl-Cys-Ser-Ser-As-Ala-Ala-OH

and the compounds resulting from replacing Ser with Phe or Phe with Ser.

A second group of compounds [Group II] according to the invention are compounds according to formula IV above, but in which the configuration at the C ^** atom is S instead of R, and the mixtures of the R and S diastereomers, as described, for example may arise in the synthetic representation. Specific types and compounds in this group also correspond to those particularly emphasized above for the "R" group (formula IV).

• lle-Asp-Glu-OH
• Asp-Glu-OH
• Spezifische Verbindungen sind z.B.
• N-Stearoyl-S-[2(R),3-Dipalmitoyloxypropyl]-Cys-Ser-Ser-Asn-Ala-Glu-lle-Asp-Glu-OH
• N-Palmitoyl-S-[2(R),3-Dipalmitoyloxypropyl]-Cys-Ser-Ser-Asn-A)a-Glu-Ile-Asp-Glu-OH
• N-Palmitoyl-S-[2(R),3-dipalmitoyloxypropyl]-Cys-Ser-Ser-Asn-Ala-Lys-Ile-Asp-Glu-OH.

Es können erfindungsgemäss auch entsprechende Verbindungen dieses Typs mit der S-Konfiguration am C^**-Atom hergestellt werden, sowie Diastereomerengemische von S- und R-Verbindungen.A third group [Group III] of the new lipopeptides comprises the compounds according to formula (IV), in which, however, X represents an amino acid sequence with 6-10 amino acids, the first 5 preferably representing the sequences already highlighted above. Amino acids 6-10 can be any of the natural ones mentioned above, such as in particular serine, asparagine, alanine, glutamic acid, lysine, phenylalanine, and can occur in the following sequences:

• ll-Asp-Glu-OH
• Asp-Glu-OH
• Specific connections are, for example
• N-Stearoyl-S- [2 (R), 3-dipalmitoyloxypropyl] -Cys-Ser-Ser-Asn-Ala-Glu-lle-Asp-Glu-OH
• N-Palmitoyl-S- [2 (R), 3-dipalmitoyloxypropyl] -Cys-Ser-Ser-Asn-A) a-Glu-Ile-Asp-Glu-OH
• N-Palmitoyl-S- [2 (R), 3-dipalmitoyloxypropyl] -Cys-Ser-Ser-Asn-Ala-Lys-Ile-Asp-Glu-OH.

Corresponding compounds of this type with the S configuration on the C ^** atom can also be prepared according to the invention, and diastereomer mixtures of S and R compounds.

insbesondere solche mit R-Konfiguration an den C^**-Atomen, worin X die für Formel (IV) angegebene Bedeutung besitzt. Die Acylreste R,-CO- und R_2-CO- sind vorzugsweise dieselben und vor allem wiederum Palmitoyl, Stearoyl oder Oleoyl; sie können aber wie oben für die anderen Gruppen besprochen. in allen Kombinationen und/oder Variationen vorhanden sein.A fourth group [Group IV] of the lipopeptides according to the invention comprises compounds of the formula (I) in which R _{3 represents} the radical R ₁ ―CO ― O ― CH ₂ -, in which the configuration on the C ^** atoms is any , in particular also mixtures of diastereomers. Subgroups are compounds of the formula

in particular those with an R configuration on the C ^** atoms, in which X has the meaning given for formula (IV). The acyl radicals R, -CO- and R _2- CO- are preferably the same and especially again palmitoyl, stearoyl or oleoyl; however, they can be discussed for the other groups as above. be available in all combinations and / or variations.

Preferred types and compounds of this class are again those in which X has the meanings specified above.

For this group too, X is preferably an amino acid or a sequence of 2-5 amino acids, in particular one of those designated as preferred for the other three groups of lipopeptides. However, X can also represent one of the amino acid sequences with 6-10 amino acids described for the third group of lipopeptides of the present application, in particular those specifically mentioned for that group.

Specific connections are e.g. which results from the table compiled above for specific compounds of group I, when in them N-acyl-0-0-di-acyl-Cys does not mean the glycerylcysteine portion, but the erythritylcysteine residue, and which is replaced by the Serine by threonine, glutamic acid, glutamine, aspartic acid or asparagine and by lysine by glutamic acid and, if desired, simultaneously by Asn by phenylalanine compounds.

• AcGCT-Phe-Ile-Ile-Phe-Ala-OH
• AcGCT-Val-Lys-Val-Tyr-Pro-OH

und ihre Amide und Ester, insbesondere die im nachfolgend besonders hervorgehobenen.A fifth group of lipopeptides according to the present invention, of those connects d _u nts according to formula (I) is formed, in which X is a peptide chain means in which the sequence of the first 5 amino acids of those in the known murein - lipoprotein - Degradation - lipopeptides present is different, and in particular those in which R _{3 is} hydrogen, that is to say corresponding to the formula (IV) above, where R, -CO and R ₂ -CO are the acyl radicals in the combinations mentioned which are preferably emphasized in the preceding or in the following , as well as their salts and complexes. The following types of lipopeptides may be mentioned (AcGCT = acyl-glycerylcysteine part according to formula (IV)

• AcGCT-Phe-Ile-Ile-Phe-Ala-OH
• AcGCT-Val-Lys-Val-Tyr-Pro-OH

and their amides and esters, especially those highlighted below.

• AcGCT-Ala-Ile-Gly-Val-Gly-Ala-PRo-OH
• AcGCT-Ser-Ser-Asn-Ala-Glu-Ile-Asp-Glu-OH

und ihre Amide und Ester, insbesondere die im nachfolgenden hervorgehobenen.Another subgroup consists of compounds according to formula (IV), in which X represents the “unnatural” sequence of amino acids in comparison to the murein-lipoprotein degradation products. and there are 6-10 amino acids. For example, the first five amino acids have the sequence given above or any other; the following lipopeptides may be mentioned, for example, where AcGCT has the above meaning:

• AcGCT-Ala-Ile-Gly-Val-Gly-Ala-PRo-OH
• AcGCT-Ser-Ser-Asn-Ala-Glu-Ile-Asp-Glu-OH

and their amides and esters, especially those highlighted below.

Compounds of this type are preferably prepared in which the configuration on the C ^** atom of the AcGCT is R, or mixtures of R and S epimers.

In the new lipopeptides according to the present invention discussed above, the terminal carboxyl group may be in amidated form. In addition to the unsubstituted amide, there are also those which are derived from a primary or secondary amine. In particular, amides of lower aliphatic amines with 1-7 C atoms are suitable, such as methylamine, ethylamine, propylamine, butylamine, dimethylamine, diethylamine or propyl- and isopropylamine, furthermore aromatic amines, especially monocyclic, such as aniline or toluidine, araliphatic, such as Benzylamine or heterocyclic amines, such as the aminopyridines. The secondary aliphatic amines can also be ring-closed nitrogen bases, e.g. Pyrrolidine, piperidine or piperazine. Specific amides are e.g. the unsubstituted or the methyl, ethyl, dimethyl or diethylamides of all of the above-mentioned specific lipopeptides according to the present invention or such amides of the compounds described in the illustrative examples.

In the esterified terminal carboxyl groups of the new lipopeptides, the alcohol component is preferably derived from lower aliphatic alcohols with 1-7 C atoms, such as methyl, ethyl, n-propyl, isopropyl or the butyl alcohols. The esterifying alcohols can also be polyvalent, such as ethylene glycol or propylene glycol or glycerin. Also araliphatic alcohols, especially monocyclic lower aliphatic with 1-7 C atoms in the aliphatic part, such as Benzyl alcohol, or heterocyclic alcohols, such as tetrahydrofuranol or tetrahydropyranol, can be used for the esterification. Specific esters of this type of lipopeptides according to the invention are e.g. the methyl, ethyl and ethylene glycol or propylene glycol esters of all of the specific lipopeptides listed above or those described in the illustrative examples.

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

Acid addition salts are particularly pharmaceutically acceptable, non-toxic acid addition salts such as those with inorganic acids, e.g. Hydrochloric, hydrobromic, nitric, sulfuric or phosphoric acids, or with organic acids such as organic carboxylic acids e.g. Acetic acid, propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid, methylmaleic acid, fumaric acid, malic acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-amino-salicylic acid, 2-phenoxybenzoic acid, niconic acid, benzoic acid or benzoic acid Isonicotinic acid, or organic sulfonic acids, e.g. Methanesulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, ethane-1,2-disulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or naphthalene-2-sulfonic acid, as well as other acid addition salts, e.g. as intermediates, e.g. for cleaning the free compounds or in the production of other sa! zes, as well as for characterization, e.g. those with picric, picrolonic, flavian, phosphotungstic, phosphomolyde, chloroplatinic, pure corner or perchloric acid.

Complexes are those with metal salts, e.g. compounds formed with heavy metal salts, such as copper, zinc, iron or cobalt salts. The phosphates, pyrophosphates and polyphosphates of these metal salts are preferably used to form such complexes, optionally in combination with acidic organic substances, e.g. acidic group-containing polysaccharides, such as carboxymethyl cellulose, tannic acid, polyglutamic acid or partially hydrolyzed gelatin, and also alkali metal polyphosphates such as e.g. "Calgon N", "Calgon 322", "Calgon 188" or "Plyron B 12".

The compounds of the present invention according to formula (1) above, their salts and complexes and mixtures have valuable pharmacological properties, in particular a pronounced immunopotentiating effect. The compounds in the dose range of 0.5-320 _{/ l} g / ml stimulate the proliferation of B lymphocytes, determined in vitro by means of thymidine incorporation, 20- to 50-fold compared to non-stimulated control lymphocytes. The extent of the stimulation corresponds to that which is achieved with the most effective known B cell mitogens [dextran sulfate, E. coli lipopolysaccharide, PPD (purified protein derivative)], with the new compounds of the present application also high concentrations not being lymphocytotoxic Act.

The new compounds of formula (1), their salts and complexes and mixtures are also able to induce the formation of antibody-producing cells in concentrations of 0.3-60 _{/ l} g / ml in spleen cell cultures of normal mice (increase in 19S- plaque-forming cells by a factor of 20 to 50 above the control value (in the absence of the stimulating substances): For example, specific antibodies against sheep erythrocytes are formed in the presence of the compounds mentioned, without adding sheep erythrocytes to the cultures for immunization. On the other hand, the substances mentioned in the same concentration range can also increase the immunological reactivity of T cell-depleted spleen cell cultures (from congenitally athymic nu / nu mice) compared to a normally thymus-dependent antigen (sheep erythrocytes) (factor 10 to 40 compared to untreated control cultures). However, the compounds mentioned not only directly or indirectly induce proliferation and synthesis performance of B-lymphocytes (ie of potentially antibody-forming cells) in vitro, but also effects on T-lymphocytes (which include regulatory active helper and suppressor cells and cytotoxic effector cells ), conveyed. For example, the compounds mentioned in a concentration range of 10-100 pg / ml can significantly (up to 10-fold) potentiate the reactivity of cortisone-resistant thymus cells with allogeneic irradiated stimulator lymphocytes.

The above-mentioned effects are probably caused indirectly by the fact that the lipopeptides activate macrophages, which in turn promote the reactivity of T and B lymphocytes. In fact, it can be shown that the compounds mentioned release large amounts of "colony stimulating activity" (CSA) from mouse macrophages even in extremely low concentrations (0.01-10 µg / ml) (induction of up to 150-200 colonies within 7 days from 10 ⁵ mouse bone marrow cells, after adding 20% supernatant from macrophage cultures incubated with substance for 24 hours, compared to 0-5 colonies when adding supernatants from untreated macrophage cultures). CSA is a biological mediator that is necessary for the differentiation of bone marrow stem cells into macrophages and polymorphonuclear leucocytes. The compounds mentioned thus bring about an increased replenishment of cells which are of central importance for the non-specific resistance and for the induction, amplification and expression of specific (lymphocyte-mediated) immune reactions.

• NMRI Mäuse werden dirch intraperitoneale Injektion von 10 µg präzipitatfreiem BSA am Tag 0 immunisiert. 9,15 und 29 Tage später werden Serumproben entnommen und auf ihren Gehalt an a_nti-BSA-Antikörpern mit einer passiven Haemagglutinationstechnik untersucht. In der verwendeten Dosis ist lösliches BSA für die Empfängertiere subimmunogen, d.h. es vermag keine oder nur eine ganz geringfügige Produktion von Antikörpern auszulösen. Zusätzliche Behandlung der Mäuse mit gewissen immunpotenzierenden Stoffen vor oder nach der Antigengabe führt zu einem Anstieg der Antikörpertiter im Serum. Der Effekt der Behandlung wird durch den erreichten Scorewert, d.h. durch die Summe der log2 Titerdifferenzen an den drei Blutungstagen ausgedrückt.

The immunopotentiating effect of the new compounds of the formula (1), their salts or complexes or mixtures can also be demonstrated in vivo: for example, the injection of a lipopeptide according to the invention leads to a high increase in the CSA concentration in the serum within 3-9 hours (up to 120 colonies per 10 ⁵ mouse bone marrow cells after adding chloroform extracted serum [5% final concentration] compared to 0-5 colonies in untreated animals). Accordingly, the administration of the same compounds in vivo significantly increases the antibody-producing ability of mice:

• NMRI mice are injected intraperitoneally with 10 µg of precipitate-free BSA on day 0 immunized. 9,15 and 29 days later, serum samples are taken and analyzed for their content of a _n ti-BSA antibodies with a passive Haemagglutinationstechnik. In the dose used, soluble BSA is subimmunogenic for the recipient animals, ie it is unable to produce any or only a very slight production of antibodies. Additional treatment of the mice with certain immunopotentiating substances before or after the antigen administration leads to an increase in the antibody titer in the serum. The effect of the treatment is expressed by the score achieved, ie by the sum of the log2 titer differences on the three bleeding days.

In this test, the compounds of the formula (I), their salts or complexes or mixtures are capable of antibody production against five days in a row after intraperitoneal or subcutaneous administration of 0.03-3 mg / kg animal before or after immunization with BSA To significantly increase BSA.

• Während Sensibilisierung von Meerschweinchen mit BSA in inkomplettem Freund'schen Adjuvans nur zu humoraler Antikörperbildung führt, induziert die Beimischung der Lipopeptide gemäss der vorliegenden Erfindung in einem Dosisbereich von 1-15 pg zur wässerigen Phase der Antigen-Oelemulsion Spättyp-Ueberempfindlichkeit gegenüber BSA: 3 Wochen nach Immunisierung führt die intrakutane Injektion von BSA bei diesen Tieren zu einer lokalen Entzündungsreaktion mit Erythem und Verdickung der Haut, die innert 24 bis 48 Stunden ihr Maximum erreicht. Diese Spättyp-Reaktionen entsprechen quantitativ und qualitativ denjenigen, die üblicherweise durch Immunisierung mit BSA in komplettem Freund' schem Adjuvans (d.h. mit Zusatz von Mykobakterien) erhalten werden. Die EDso-Werte (benötigte µg/Tier zur Induktion einer Differenz des Reaktionsvolumens (Erythemfläche x Hautdickenzunahme) bei behandelten und unbehandelten Tieren von 200 pi, 24 Stunden nach Auslösung) betragen 2-3 ug.

Manifestations of cell-mediated immunity can also be potentiated in vivo by the compounds mentioned:

• While sensitization of guinea pigs with BSA in incomplete Freund's adjuvant only leads to humoral antibody formation, the addition of the lipopeptides according to the present invention in a dose range of 1-15 pg to the aqueous phase of the antigen-oil emulsion induces late-type hypersensitivity to BSA: 3 weeks after immunization, the intracutaneous injection of BSA in these animals leads to a local inflammatory reaction with erythema and thickening of the skin, which reaches its maximum within 24 to 48 hours. These late-type reactions correspond quantitatively and qualitatively to those which are usually obtained by immunization with BSA in a complete Freund's adjuvant (ie with the addition of mycobacteria). The ED 50 values (µg / animal required to induce a difference in the reaction volume (erythema area x increase in skin thickness) in treated and untreated animals of 200 pi, 24 hours after triggering) are 2-3 µg.

The lipopeptides according to the present invention are also not very toxic: 5 times intraperitoneal application in a dose of 10 mg / kg / day on five consecutive days was apparently tolerated without symptoms by mice. Since the doses required for immunostimulation are very small, the therapeutic breadth of the new compound is very large.

The new lipopeptides according to the present invention can thus significantly increase the cellular and especially the humoral immunity, both in a mixture with the antigen itself (adjuvant effect in the narrower sense) and with a supply that is separate in time and place from the antigen injection (systemic immunopotentiation).

The new lipopeptides according to the present invention can thus be used as adjuvants in admixture with vaccines to improve the vaccination success and to improve the protection against infection mediated by humoral antibodies and / or cellular immunity against bacterial, viral or parasitic pathogens.

Finally, the compounds described, in a mixture with various antigens, are suitable as adjuvants in the experimental and industrial production of antisera for therapy and diagnosis and in the induction of immunologically activated lymphocyte populations for cell transfer processes.

In addition, the new lipopeptides can also be used without the simultaneous supply of antigen to promote immune reactions in humans and animals that are already taking place. The compounds are therefore particularly suitable for stimulating the physical defense, e.g. in chronic and acute infections or in selective (antigen-specific) immunological defects, as well as in congenital, but also in acquired general (ie non-antigen-specific) immunoligical defect states, such as those in old age, in the course of severe primary diseases and especially after therapy with ionizing radiation or with immunosuppressive hormones occur. The substances mentioned can therefore preferably also be administered in combination with antibiotics, chemotherapeutic agents or other therapeutic methods in order to counteract immunological damage. Finally, the substances described are also suitable for the general prophylaxis of infectious diseases in humans and animals.

• a) dass man in einer Verbindung der Formel
  
  worin R₁, R₂ und R₃ die für Formel (I) angegebene Bedeutung haben und Y eine X in der Formel (I) entsprechende Aminosäure oder Aminosäurensequenz darstellt, worin jedoch mindestens eine der die Aminosäuren substituierenden hydrophilen Gruppen und/oder die terminale Carboxylgruppe durch eine unter neutralen oder milden sauren Bedingungen abspaltbare Schutzgruppe geschützt ist, oder in einem Salz einer solchen Verbindung, die Schutzgruppe(n) abspaltet, oder
• b) dass man eine Verbindung der Formel
  
  worin R₁, R₂ und R₃ die für Formel (I) angegebene Bedeutung haben und W = OH oder eine Aminosäure oder eine unvollständige Sequenz von Aminosäuren gemäss X in Formel (I) darstellt, mit einer Verbindung der Formel
  
  wo Z₁ eine der Gruppe X in Formel (I) entsprechende Gruppe bzw. eine zur genannten Aminosäure oder unvollständigen Sequenz von Aminosäuren gemäss X komplementären Aminosäuresequenz bzw. Aminosäure bedeutet, in welchen Sequenzen jedoch keine freien Aminogruppen vorhanden sind, oder einem Salz derselben, umsetzt, oder
• c) eine Verbindung der Formel
  
  , worin R₂ die für Formel (I) Angegebene Bedeutung besitzt und R₃ Wasserstoff oder die Gruppe HQ-CH₂ darstellt, und Z₂ eine X in Formel (I) entsprechende Gruppe darstellt, worin jedoch keine freien Hydroxygruppen vorhanden sing, oder ein Salz dieser Verbindung, in an sich bekannter Weise acyliert, und, wenn erwünscht, in erhaltenen Verbindungen mit freier terminaler Carboxylgruppe dieselbe in eine Amidgruppe oder Estergruppe überführt, und/oder die Verbindungen in ihre Salze oder Komplexe überführt.

The new lipopeptides can be produced by methods known per se. In a preferred process, the compounds of the formula (I), their diastereomeric mixtures and their salts and complexes are prepared by

• a) that one in a compound of the formula
  
  wherein R ₁ , R ₂ and R _{3 have} the meaning given for formula (I) and Y represents an amino acid or amino acid sequence corresponding to X in formula (I), but in which at least one of the hydrophilic groups substituting the amino acids and / or the terminal carboxyl group is protected by a protective group which can be split off under neutral or mild acidic conditions, or in a salt of such a compound which closes off protective group (s), or
• b) that you have a compound of formula
  
  wherein R ₁ , R ₂ and R _{3 have} the meaning given for formula (I) and W = OH or an amino acid or an incomplete sequence of amino acids according to X in formula (I), with a compound of the formula
  
  where Z ₁ denotes a group corresponding to group X in formula (I) or an amino acid sequence or amino acid complementary to the amino acid or incomplete sequence of amino acids according to X, but in which sequences no free amino groups are present, or a salt thereof , or
• c) a compound of the formula
  
  , in which R _{2 has} the meaning given for formula (I) and R _{3 represents} hydrogen or the group HQ-CH ₂ , and Z _{2 represents} a group corresponding to X in formula (I), but in which no free hydroxyl groups are present, or a Salt of this compound, acylated in a manner known per se, and, if desired, converted the resulting compounds having a free terminal carboxyl group into an amide group or ester group, and / or converting the compounds into their salts or complexes.

The protective groups in the process according to variant a) are particularly known from the synthesis of peptides.

For example, Protecting groups for amino groups, acyl or aralkyl groups such as formyl, trifluoroacetyl, phthaloyl, benzenesulfonyl, p-toluenesulfonyl, o-nitrophenylsulfenyl, 2,4-dinitrophenylsulfenyl groups (these sulfenyl groups can also be formed by the action of nucleophilic reagents, eg sulfites, e.g. sulfites are split off), optionally substituted, such as benzyl groups, or diphenyl or triphenylmethyl groups substituted by lower alkoxy groups, especially o- or p-methoxy groups, or groups derived from carbonic acid, such as optionally in the aromatic rings, e.g. by halogen atoms such as chlorine or bromine, nitro groups, lower alkyl or lower alkoxy groups or coloring groups, e.g. Arylmethyloxycarbonyl groups substituted in azo groups, in which the methylene group can be substituted by a further aryl radical and / or one or optionally two lower alkyl radicals, such as benzyl, benzhydryl or 2-phenyl-isopropyloxycarbonyl groups, e.g. Carbobenzoxy, p-bromo- or p-chlorocarbobenzoxy, p-nitrocarbobenzoxy or p-methoxycarbobenzoxy, p-phenylazo-benzyloxycarbonyl and p- (p'-methoxy-phenylazo) -benzyloxycarbonyl, 2-tolyl-isopropyloxycarbonyl and especially 2- (p- Biphenylyl) isopropyloxycarbonyl, and aliphatic oxycarbonyl groups such as adamantyloxycarbonyl, cyclopentyloxycarbonyl, trichloroethyloxycarbonyl, tert. Amyloxycarbonyl or especially tert-butyloxycarbonyl.

The amino groups can also be formed by formation of enamines obtained by reacting the amino group with 1,3-diketones, e.g. Benzoylacetone, acetylacetone or dimedone.

Carboxyl groups are protected, for example, by amide or hydrazide formation or by esterification. The amide and hydrazide groups can be optionally substituted, the amide group e.g. by the 3,4-dimethoxybenzyl or bis- (p-methoxyphenyl) methyl group, the hydrazide group e.g. by the carbobenzoxy group, the trichloroethyloxycarbonyl group, the trifluoroacetyl group, the trityl group, the tert-butyloxycarbonyl group or the 2- (p-biphenylyl) isopropyloxycarbonyl group. Suitable for esterification are e.g. lower optionally substituted alkanols such as methanol, ethanol, cyanomethyl alcohol, benzoylmethyl alcohol or in particular tert-butanol, furthermore aralkanols such as aryl lower alkanols, e.g. benzyl alcohols or benzhydrols such as benzhydrol, p-nitrobenzyl alcohol, p-methoxybenzyl alcohol, 2,4,6-trimethylbenzyl alcohol optionally substituted by lower alkyl or lower alkoxy groups or halogen atoms, phenols optionally substituted by electron-withdrawing substituents and thiophenols such as thiophenol, thiocresol, p-nitronol 4,5- and 2,4,6-trichlorophenol, pentachlorophenol, p-nitrophenol, 2,4-dinitrophenol, p-cyanophenol or p-methanesulfonylphenol, further, for example N-hydroxysuccinimide, N-hydroxyphthalimide, N-hydroxypiperidine, 8-hydroxyquinoline.

The hydroxyl groups of the serine and threonine residues can e.g. be protected by esterification or etherification.

Suitable acyl radicals in the esterification are, in particular, radicals derived from carbonic acid, such as benzoyloxycarbonyl or ethyloxycarbonyl. Groups suitable for etherification are, for example, benzyl, tetrahydropyranyl or tert-butyl radicals. Furthermore, suitable for protecting the hydroxyl groups the in Ber. 100 (1967), 3838-3849, describes 2,2,2-trifluoro-1-tert-butyloxycarbonylamino or -1-benzyloxycarbonylaminoethyl groups (Weygand).

The tert-butyl ester group or the benzhydrol group is preferably used to protect the carboxyl group of the side chains and optionally the terminal carboxyl group, the tert-butyloxycarbonyl group to protect the amino groups of the side chains, for the hydroxyl groups of serine or threonine, the tert-butyl ether group, and if desired, the 2,2,2-trifluoro-1-tert-butyloxycarbonylaminoethyl group to protect the imino group of the histidine.

The process-related removal of the protective groups with acidic agents under mild conditions is carried out e.g. in a manner known from peptide chemistry, e.g. by treatment with trifluoroacetic acid.

worin R^l, R² und R³ je einen Kohlenwasserstoffrest bedeutet, wobei die Reste untereinander auch durch eine einfache C-C Bindung verknüpft sein können, insbesondere Alkylreste mit 1-5 C Atomen. Schutzgruppen dieses Typs sind z.B. die 2-(Dimethyl-tert.-butylsilyl)-äthyl-, die 2-(Dibutyl-methyl-silyl)-äthyl- und vor allem die 2-(Trimethylsilyl)-äthylgruppe. Obwohl diese Schutzgruppen auch basisch abgespalten werden können, ist besonders ihre Abspaltbarkeit unter neutralen Bedingungen, und zwar durch Einwirking eines Salzes der Fluorwasserstoffsäure, von Interesse. Die Abspaltung der Schutzgruppe wird vorteilhaft in einem aprotischen organischen Lösungsmittel vorgenommen; vorzugsweise wird dabei die Anwesenheit von solchen Lösungsmitteln vermieden, welche das Fluorid-Anion zu solvatisieren vermögen, wie Wasser oder niederaliphatische Alkohole.A special protective group for carboxyl groups which can be split off under neutral conditions is the group of the general formula described, for example, in German Offenlegungsschrift 27 06 490

where R ¹ , R ² and R ³ each represent a hydrocarbon radical, where the radicals can also be linked to one another by a simple CC bond, in particular alkyl radicals having 1-5 C atoms. Protecting groups of this type are, for example, the 2- (dimethyl-tert-butylsilyl) ethyl, the 2- (dibutyl-methyl-silyl) ethyl and especially the 2- (trimethylsilyl) ethyl group. Although these protective groups can also be split off under basic conditions, their splittability under neutral conditions is of particular interest, namely by the action of a salt of hydrofluoric acid. The protective group is advantageously removed in an aprotic organic solvent; the presence of solvents which are able to solvate the fluoride anion, such as water or lower aliphatic alcohols, is preferably avoided.

The condensation according to variant b) of the compound of formula (VII) with the compound of formula (VIII) takes place e.g. in such a way that the compound (VII) is reacted with (VIII) in the form of the activated carboxylic acid, or in that the acid (VII) is reacted with the compound (VIII), the amino group of which is present in activated form. If appropriate, the compounds (VII) or (VIII) are used for the condensation in the form of their salts.

The carboxyl group of the compound (VII) can be converted, for example, into an acid azide, anhydride, imidazolide, isoxazolide or an activated ester, such as cyanomethyl ester, carboxymethyl ester, thiophenyl ester, p-nitrothiophenyl ester, thiocresyl ester, p-methanesulfonylphenyl ester, p-nitro , 4-dinitrophenyl ester, 2,4,5- or 2,4,6-trichlorophenyl ester, pentachlorophenyl ester, N-hydroxysuccinimide ester, N-hydroxyphthalimide ester, 8-hydroxyquinoline ester, 2-hydroxy-1,2-dihydro-1-carbäthoxy-quinoline- esters, N-hydroxypiperidine esters or an enol ester obtained with N-ethyl-5-phenyl-isoxazolium-3-sulfonate [Woodward Reagent], or by reaction using a carbodiimide (optionally with the addition of N-hydroxysuccinimide) or an unsubstituted or For example, 1-hydroxybenzotriazole, 3-hydroxy-4-oxo-3,4-d1hydrobenzo [d] -1,2,3-triazine or N, N ^I- carbonyldiimidazole, can be activated by halogen, methyl or methoxy.

The amino group of the compound (VIII) can be activated, for example, by reaction with a phosphite.

The most common methods of condensation are: the Weygand-Wünsch method (carbodiimide in the presence of N-hydroxysuccinimide), the azide method, the N-carboxyanhydride or N-thiocarboxyanhydride method, the activated ester method and the anhydride method. In particular, these condensations can be carried out using the Merrifield method.

It is also possible to carry out the condensation of the compound (VII) by the methods just mentioned with a compound of the formula (VIII) in which the terminal carboxyl group is protected by a protective group other than an amide or ester group as defined for X. , such as one of the above, is blocked, and / or one or more of the hydrophilic groups present are in protected form, as described above. This leads to condensation products which belong to the group of starting materials of the formula (VI) used for variant a) described above, which are likewise the subject of the present invention.

According to variant c) above, the free hydroxyl groups in the glyceryl or erythrityl radical are acylated in compounds of the formula (IX) which may not contain any free hydroxyl groups in part Z _Z. This acylation can be carried out in a manner known per se, for example by reaction with a reactive functional derivative of the acid corresponding to the radical to be introduced, such as the anhydride or an acid halide, preferably in the presence of a tertiary base such as pyridine or collidine.

This process is particularly suitable for the production of process products according to formula I in which the acyl residues in the glyceryl or. Erythrityl residue and the cysteine residue are different.

This variant is used for compounds of the formula (IX) in which, in part Z _2, the terminal carboxyl group is protected by other protective groups, in addition to being protected by an amide or ester radical, as provided for X, and / or one or more of the existing hydrophilic groups in a protected form, as described above, there are also starting compounds for variant a) above.

In the lipopeptides according to formula I obtained by any of the described process variants, in which a free terminal carboxyl group is present in part X, the same can be used in a manner known per se, e.g. be converted into the amide or ester group by one of the methods customary in peptide chemistry.

An N-acyl or N, O, O-triacyl-S- (2,3-dihydroxypropyl) cysteine to be used as starting material according to formula (VII), provided that W = OH and R ₃ = H, can be found in the following Are obtained as follows: starting from 1,2,5,6-D-mannitol diacetonide, 2-O, 3-O-isopropyl-idene-D-glycerinaldehyde is obtained by periodate oxidation, the aldehyde group is reduced to the carbinol group and esterified with p-toluenesulfonic acid . The 1-0-tosyl-2 (R) -O, 3-0-isopropylidene glycerol thus obtained is condensed with N-acyl- (R) -cysteine in the presence of a basic agent, for example potassium carbonate, and the N-acyl- S- [2 (R), 3-isopropylenedioxy-propyll- (R) -cysteine, which is acidified, for example with acetic acid. The N-acyl-S- [2 (R), 3-dihydroxypropyll- (R) -cysteine thus obtained can, if desired, be in the 2- and 3-position of the glycerol portion, preferably with temporary protection of the carboxyl group, for example by means of the Trimethylsilylethyl group described above, or preferably the benzhydryl ester group, can be further acylated.

The preparation of an N-acyl or N, 0,0,0-tetraacyl-S- (2,3,4-trihydroxybutyl) cysteine to be used as the starting material according to formula (VII) can be carried out analogously to the method of Example 7.

The amino acids or peptides to be used for the production of the new lipopeptides according to the process are known or can be prepared by methods known per se.

The above starting compounds (VII) for variant b) in the event that W = OH, and their diastereomeric mixtures, salts and complexes themselves also have an immunopotentiating effect, these in vitro in the same dose range as for the compounds of the formula and their derivatives specified occurs. In vivo, the compounds mentioned show an increase in the antibody titer in the serum in the dose range of 1.0 to 3 mg / kg animal in the test for potentiating the humoral immunity in the intraperitoneal administration 5 days before the antigen administration.

These compounds can be obtained by acylating a compound of the formula (IX) in which, however, a hydroxyl group is present instead of Z ₂ , as described above for variant c). The preparation of the starting materials required for this, ie the glyceryl or erithrityl-N-acylcysteine, is described above. Compounds of the formula (IX) in which Z _{2 has} a meaning corresponding to the group X in formula (I) and but in which no free hydroxyl groups are present can be reacted with a compound by reacting the N-acyl-cysteine derivatives just mentioned of the formula NH _2- Z ₂ according to the same principles as specified for variant b) of the general process for the preparation of the lipopeptides according to the invention described above.

The lipopeptides obtained can be converted into their salts in a manner known per se, e.g. by reacting acidic compounds obtained with alkali or alkaline earth metal hydroxides or basic compounds obtained with acids.

As a result of the close relationship between the new compounds in free form and in the form of their salts and complexes, the corresponding salts are to be understood in the preceding and following as appropriate and expediently, if appropriate, the corresponding salts.

Isomer mixtures obtained can be separated in a known manner on the basis of the physico-chemical differences in the constituents, for example by chromatography and / or fractional crystallization. The more effective of the isomers is advantageously isolated.

The methods described above are e.g. carried out according to methods known per se, in the absence or preferably in the presence of diluents or solvents, if necessary, with cooling or heating, under elevated pressure and / or in an inert gas, such as a nitrogen atmosphere. Taking into account all substituents in the molecule, if necessary, especially in the presence of easily hydrolyzable O-acyl residues, particularly gentle reaction conditions, such as short reaction times, use of mild acidic agents in low concentration, stoichiometric proportions, choice of suitable catalysts, solvents, temperature and / or printing conditions.

The invention also relates to those embodiments of the process in which one starts from a compound obtainable as an intermediate at any stage of the process and carries out the missing process steps, or terminates the process at any stage, or forms a source material under the reaction conditions or in the form of a reaction capable derivative or salt used. The starting materials used are preferably those which, according to the process, lead to the compounds described above as being particularly valuable.

The present invention also relates to pharmaceutical preparations which contain the described new lipopeptides according to the invention, both those of the formula (I) and those of the formula (VII), their mixtures, salts or complexes. In the pharmaceuticals according to the invention Table preparations are those for enteral, such as oral or rectal, as well as parenteral administration to warm-blooded animals, which contain the pharmacologically active substance alone or together with a pharmaceutically usable carrier material. The dosage of the active ingredient depends on the warm-blooded species, the age and the individual condition, as well as on the mode of administration.

The new pharmaceutical preparations contain from about 10% to about 95%, preferably from about 20% to about 90% of the active ingredient. Pharmaceutical preparations according to the invention can e.g. in unit dose form, such as coated tablets, tablets, capsules, suppositories or ampoules.

The pharmaceutical preparations of the present invention are manufactured in a manner known per se, e.g. manufactured using conventional mixing, granulating, coating, solution or lyophilization processes.

Suitable carriers are in particular fillers such as sugar, e.g. Lactose, sucrose, mannitol or sorbitol, cellulose preparations and / or calcium phosphates, e.g. Tricalcium phosphate or calcium hydrogen phosphate, further binders such as starch paste using e.g. of corn, wheat, rice or potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose and / or polyvinyl pyrrolidone, and / or, if desired, disintegrants, such as the above-mentioned starches, also carboxymethyl starch, cross-linked polyvinyl pyrrolidone, agar, Alginic acid or a salt thereof, such as sodium alginate, are primarily flow regulators and lubricants, for example Silicic acid, talc, stearic acid or salts thereof, such as magnesium or calcium stearate, and / or polyethylene glycol. Dragee cores are provided with suitable, possibly enteric coatings, whereby one of the things Concentrated sugar solutions, which may contain arabic gum, talc, polyvinylpyrrolidone, polyethylene glycol and / or titanium dioxide, lacquer solutions in suitable organic solvents or solvent mixtures or, for the production of gastric juice-resistant coatings, solutions of suitable cellulose preparations, such as acetyl cellulose phthalate or hydroxypropylmethyl cellulose phthalate. Colorants or pigments, e.g. for identification or for labeling different doses of active ingredient.

The following examples illustrate the invention described above; however, they are not intended to limit their scope in any way. Temperatures are given in degrees Celsius. The abbreviations used mean:

• System 3: Essigester-Pyridin-Wasser (65:20:15)
• System 157: Chloroform-Methanol-Wasser-Eisessig (70:42:10:0.5)
• System 157c: Chloroform-Methanol-Wasser-Eisessig (75:25:5:0.5)

In DC, silica gel is used as the adsorbent and the following systems as the eluent:

• System 3: ethyl acetate-pyridine-water (65:20:15)
• System 157: chloroform-methanol-water-glacial acetic acid (70: 42: 10: 0.5)
• System 157c: chloroform-methanol-water-glacial acetic acid (75: 25: 5: 0.5)

example 1

1.13 g [N-Palmitoyl-S-2 (R), 3-dipalmitoyloxypropyl] -Cys-Ser (Bu ^t ) -Ser (Bu ^t ) -Asn-OBut are in 5 ml 90 percent. Trifluoroacetic acid was added and the solution was concentrated to about 2 ml after 45 minutes at 20 °. The product is precipitated by adding 75 ml of peroxide-free ether, filtered off and dried over potassium hydroxide. The lipopeptide [N-palmitoyl-S-2 (R), 3-dipalmitoyloxy-, propyl] -Cys-Ser-Ser-Asn-OH thus obtained shows the following RF values in a thin layer chromatogram on silica gel: Rf (chloroform-methanol-glacial acetic acid -Water 79: 25: 0.5: 4.5) = 0.25.

0.89 A [N-Palmitoyl-S-2 (R), 3-dihydroxypropyl] -Cys-Ser (Bu ^t ) -Ser (Bu ^t ) -Asn-OBu ^t are taken up in 10 ml pyridine, 0.725 ml palmitic acid chloride is added and the Solution kept at 45 ° for 24 hours. Then 200 ml of chloroform are added and the solution is extracted with 1N citric acid, 1N sodium bicarbonate and water, the organic phase is dried over sodium sulfate and the solvent is evaporated off. The residue is purified by chromatography on silica gel in the chloroform or chloroform-methanol (98: 2) system. The lipopeptide [N-palmitoyl-S-2 (R), 3-dipalmitoyloxypropyl) -Cys-Ser (Bu ^t ) -Ser (Bu ^t ) -Asn-OBu ^t shows in the system chloroform-methanol (9: 1) on silica gel an Rf value of 0.75.

To 0.90 g of [N-palmitoyl-S-2 (R), 3-dihydroxypropyl] cysteine and 0.99 g H-Ser (Bu ^t) -Ser (Bu ^t) -Asn-OBu ^t in 20 ml of dimethylformamide 0.472 g of dicyclohexylcarbodiimide and 0.309 A of N-hydroxybenzotriazole are added at 0 °. After 2 hours at 0 ° and 15 hours at 20 °, the mixture is filtered off and the filtrate is evaporated. The residue is purified by chromatography on a column of silica gel in the Chloroform-methanol system (98-2). In the thin layer chromatogram on silica gel, the lipopeptide derivative thus obtained [N-palmitoyl-S-2 (R), 3-dihydroxypropyl] -Cys-Ser (Bu ^t ) -Ser (Bu ^t ) -A _sn - _OBu t shows an Rf- Value of 0.60 (chloroform-methanol 8: 2).

6.09 g of Z-Ser (Bu ^t ) -Ser (Bu ^t ) -Asn-OBu ^t are dissolved in 60 ml of methanol and after addition of 0.5 g of Pd-carbon (10%) at room temperature for 3 hours hydrated. The catalyst is filtered off and the filtrate is evaporated. The product is a white foam. In the thin layer chromatogram on silica gel, the peptide H-Ser (Bu ^t ) -Ser (Bu ^t ) -Asn-OBu ^{t shows} an Rf value of 0.41 in the chloroform-methanol system (8: 2).

4.134 g Z-Ser (Bu ^t) -OH and 4.64 g H-Ser (Bu ^t) -Asn-OBu ^t was dissolved in 50 ml of dimethylformamide and treated at 0 ° 3.172 g of dicyclohexylcarbodiimide and 1.89 g of N-hydroxybenzotriazole were added . After 2 hours at 0 ° and 15 hours at 20 °, the precipitated dicyclohexylurea is filtered off with suction, the filtrate is evaporated, the residue is taken up in ethyl acetate and the solution is extracted with 1N citric acid, 1N sodium bicarbonate and water, dried over sodium sulfate and evaporated. The residue is recrystallized from ethyl acetate-hexane. The peptide Z-Ser (Bu ^t ) -Ser (Bu ^t ) Asn-OBu ^{t obtained} has a melting point of 96-98 °. On silica gel: Rf (chloroform-methanol 95: 5) = 0.25.

4.65 g Z-Ser (Bu ^t) -Asn-OBu ^t be in 50 ml methanol in the presence of 0.4 g of Pd-carbon hydrogenated (ig 1 O%). After filtering off the catalyst and evaporating the filtrate, the peptide H-Ser (Bu ^t ) Asn-OBu ^{t is obtained} as a white foam. On silica gel: Rf (chloroform-methanol 7: 3) = 0.48.

5.9 g of Z-Ser (Bu ^t) -OH and 3.76 g of H-Asn-OBu ^t was dissolved in 60 ml of dimethylformamide and treated at 0 ° with 3.06 g of N-hydroxybenzotriazole and 4.53 g dicyclohexylcarbodiimide was added. After 2 hours at 0 ° and 15 hours at 20 °, the mixture is filtered, the filtrate is evaporated and the residue is crystallized from ethyl acetate-petroleum ether. The peptide Z-Ser (Bu ^t ) -Asn-OBu ^{t obtained} melts at 114-115 °. [α] ²⁰ _D = + 4 ° (C = 1.5 in methanol). On silica gel: Rf (chloroform-methanol 9: 1) = 0.48.

Example 2

1.6 g of [N-palmitoyl-S-2 (R), 3-dipalmitoyloxypropyl] -Cys-Ser (Bu ^t) -Ser (Bu ^t) -Asn-Ala-OBu ^t are in 7.5 ml of 90% -Trifluoroacetic acid dissolved and the solution after 15 minutes at 20 ° concentrated to about 2 ml. The product is precipitated with 100 ml of ether, filtered and dried over sodium hydroxide. The result is a white powder with a melting point of 215-217 ° (decomposition), which is the lipopeptide [N-palmitoyl-S-2 (R), 3-dipalmotoyloxypropyl] -Cys-Ser-Ser-Asn-Ala-OH. On silica gel: Rf (157) = 0.55.

Example 3

0.95 g of [N-palmitoyl-S-2 (R), 3-dipalmotoyloxypropyl] -Cys-Ser (Bu ^t) -Ser (Bu ^t) -OBu ^t are dissolved in 4 ml of 90% strength trifluoroacetic acid. After 15 minutes at 20 °, 100 ml of ether are added, the mixture is filtered off and the residue is dried over sodium hydroxide. The lipopeptide [N-palmitoyl-S-2 (R), 3-dipalmotoyloxypropyl] Cys-Ser-Ser-OH obtained is obtained as a colorless, non-hygroscopic powder. On silica gel: Rf (chloroform-methanol-glacial acetic acid-water 79: 25: 0.5: 4.5) = 0.53.

Example 4

• 13,9 g N-Palmitoyl-(R)-cystein, 5,5 g Kaliumcarbonat und 15 g 1-0-Tosyl-2(R)-0-3-0-isopropyliden-glyzerin erhitzt man 8 Stunden bei 80° in 250 ml Aethanol unter Stickstoff. Man reinigt das nach dem Eindampfen erhaltene Reaktionsgemisch durch Chromatographie an 400 g Kieselgel Merck durch Elution, zuerst mit Chloroform/Aceton (8:2), dann mit Chloroform/Methanol (8:2) Man erhält so das Kaliumsalz des N-Palmitoyl-S-[2(R),3-isopropylidendioxypropyl]-(R)-cystein, das durch 4- stündiges Erhitzen auf 80° in 80% wässriger Essigsäure verseift wird. Man dampft zur Trockne, löst den Rückstand in Chloroform und schüttelt mit Wasser aus. Nach Trocknen und Eindampfen der Chloroform-Phase erhält man einen farblosen Rückstand, der aus Cyclohexan umkristallisiert wird und das N-Palmitoyl-S-[2(R),3-dihydroxypropyl]-(R)-cystein darstellt. Fp. 110°, [α]²⁰ _D =― 25° (c = 0.9, MeOH).

The N-palmitoyl-S-2 (R), 3-dihydroxypropyl] - (R) -cysteine used as starting material in Example 1 can be prepared as follows:

• 13.9 g of N-palmitoyl- (R) -cysteine, 5.5 g of potassium carbonate and 15 g of 1-0-tosyl-2 (R) -0-3-0-isopropylidene-glycerol are heated at 80 ° in for 8 hours 250 ml of ethanol under nitrogen. The reaction mixture obtained after evaporation is purified by chromatography on 400 g of Merck silica gel by elution, first with chloroform / acetone (8: 2), then with chloroform / methanol (8: 2). The potassium salt of N-palmitoyl-S is thus obtained - [2 (R), 3-isopropylidendioxypropyl] - (R) -cysteine, which is saponified by heating for 4 hours at 80 ° in 80% aqueous acetic acid. It is evaporated to dryness, the residue is dissolved in chloroform and shaken out with water. After drying and evaporating the chloroform phase, a colorless residue is obtained, which is recrystallized from cyclohexane and which is N-palmitoyl-S- [2 (R), 3-dihydroxypropyl] - (R) -cysteine. Mp 110 °, [α] ²⁰ _D = - 25 ° (c = 0.9, MeOH).

• 45 g (R)-Cystein (0,37 mol) suspendiert man in 350 ml Pyridin und tropft unter gutem Rühren und in einer Stickstoffatomosphäre bei Raumtemperatur eine Lösung von 140 ml (1,67 Aequivalente) Palmitinsäurechlorid in 550 ml Methylenchlorid zu. Nach 20-stündigem Rühren säuert man mit 2N-Salzsäure an und verteilt zwischen Chloroform und Wasser. Nach Trocknen und Eindampfen der Chloroform-Phase erhält man ein Kristallgemisch, bestehend aus etwa einem Drittel Dipalmitoylcystein und etwa zwei Drittel Monopalmitoylcystein. Man extrahiert dreimal mit 1 Liter heissem Aceton, aus dem ein 1 : 1-Gemisch von Di- und Mono-Palmitoylcystein auskristallisiert. Aus der Acetonlösung erhält man durch Eindampfen N-Palmitoylcystein, das aus Benzin umkristallisiert wird. Fp 65­67°. [α]²⁰ _D = + 1 ° (c = 0,8; Methanol).

The N-palmitoyl- (R) -cysteine can be produced as follows:

• 45 g of (R) -cysteine (0.37 mol) are suspended in 350 ml of pyridine and a solution of 140 ml (1.67 equivalents) of palmitic acid chloride in 550 ml of methylene chloride is added dropwise with good stirring and in a nitrogen atmosphere at room temperature. After stirring for 20 hours, acidify with 2N hydrochloric acid and partition between chloroform and water. After drying and evaporating the chloroform phase, a crystal mixture consisting of about a third of dipalmitoylcysteine and about two thirds of monopalmitoylcysteine is obtained. It is extracted three times with 1 liter of hot acetone, from which a 1: 1 mixture of di- and mono-palmitoylcysteine crystallizes out. Evaporation gives N-palmitoylcysteine from the acetone solution, which is recrystallized from gasoline. Mp 6567 °. [α] ²⁰ _D = + 1 ° (c = 0.8; methanol).

The mixtures of di- and mono-palmitoyl-cysteine are dissolved in methanolic solution with the addition of an aqueous solution of 12 g sodium sulfide with 2 ml conc. Sodium hydroxide solution treated for 15 minutes at room temperature. After evaporating off the methanol and acidifying the residue with 2N Hydrochloric acid gives the remaining N-palmitoyl-cysteine by chloroform extraction, which is also recrystallized from gasoline.

Example 5

0.80 g N-Palmitoyl-S- [2 (R), 3-dipalmitoyloxypropyl] - (R) -cysteine and 0.67 g H-Ser (Bu ^t ) -Ser (Bu ^t ) -Phe-Ala-Glu (OBu ^t ) ₂ are taken up in 10 ml of dimethylformamide and 0.22 g of dicyclohexylcarbodiimide and 0.16 g of N-hydroxybenzotriazole are added. By briefly heating to 40 ° a clear solution is obtained which solidifies to a gelatin after about an hour at 20 °. The latter is brought back into solution after 24 hours by heating to 40 ° and the N-palmitoyl-S- [2 (R), 3-dipalmitoyloxypropyl] -Cys-Ser (Bu ^t ) -Ser (Bu ^t ) -Phe- Ala-Glu (OBu ^t ) ₂ precipitated with water. The product is purified by chromatography on silica gel in a chloroform or chloroform-methanol 98: 2 system and then has an Rf value of 0.64 in chloroform-methanol-water 95: 5.

540 mg of the compound obtained in this way are dissolved in 30 ml of 90% trifluoroacetic acid and the solution is concentrated to about half after 30 minutes at 20 ° and the N-palmitoyl-S- [2 (R), 3-dipalmitoyloxypropyl] - with petroleum ether. Cys-Ser-Ser-Phe-Ala-Glu-OH precipitated. The compound is centrifuged off, washed three times with petroleum ether and dried over potassium hydroxide. It represents a white powder with the Rf value (System 157) = 0.63.

• N-Palmitoyl-S-[2(R,S),3-dipalmitoyloxypropyl]-Cys-Phe-Phe-Asn-Ala-Lys-OH, Rf (System 157 c) = 0,18;
• N-Myristoyl-S-[2(R),3-dipalmitoyloxypropyll-Cys-Glu-Gln-Asn-Ala-Lys-OH, Rf (System 157 c) = 0,12;
• N-Lauroyl-S-[2(R),3-dipalmitoyloxypropyl]-Cys-Ser-Ser-Asn-Ala-Glu-OH, Rf (System 157 c)=0,20;
• N-Stearoyl-S-[2(R),3-dipalmitoyloxypropyl]-Cys-Ser-Ser-Asn-Ala-Ala-OH, Rf

The following lipopeptides are produced in an analogous manner:

• N-Palmitoyl-S- [2 (R, S), 3-dipalmitoyloxypropyl] -Cys-Phe-Phe-Asn-Ala-Lys-OH, Rf (System 157 c) = 0.18;
• N-myristoyl-S- [2 (R), 3-dipalmitoyloxypropyll-Cys-Glu-Gln-Asn-Ala-Lys-OH, Rf (System 157 c) = 0.12;
• N-Lauroyl-S- [2 (R), 3-dipalmitoyloxypropyl] -Cys-Ser-Ser-Asn-Ala-Glu-OH, Rf (System 157 c) = 0.20;
• N-stearoyl-S- [2 (R), 3-dipalmitoyloxypropyl] -Cys-Ser-Ser-Asn-Ala-Ala-OH, Rf

• 17,22 g Z-Ala-ONp, 14,8 g HCI. HGlu(OBu^t)₂ und 6,3 ml N-Ethylmorpholin werden in 40 ml Dimethylformamid über Nachtbei 20° stehen gelassen. Die Lösung wird eingedampft, der Rückstand in Essigester aufgenommen und mit 1-N-Natriumbicarbonat und Wasser extrahiert.

The peptide described above with the protective groups mentioned and which is used as a starting material for the condensation with N-palmitoyl-S- [2 (R), 3-dipalmitoyloxypropyl] - (R) -cysteine can be prepared as follows:

• 17.22 g Z-Ala-ONp, 14.8 g HCl. HGlu (OBu ^t ) ₂ and 6.3 ml of N-ethylmorpholine are left in 40 ml of dimethylformamide overnight at 20 °. The solution is evaporated, the residue is taken up in ethyl acetate and extracted with 1N sodium bicarbonate and water.

The ethyl acetate solution is evaporated and the foam left is the Z-Ala-Glu (OBu ^t ) ₂ with the Rf value 0.60 (in chloroform-methanol 95: 5), which is directly processed further: 8.5 g are made in 60 ml of methanol dissolved and after adding 0.8 mg of Pd carbon (10%) hydrogenated at 20 ° for 2 hours. The catalyst is filtered off and the filtrate is evaporated. The H-Ala-Glu (OBu ^t ) _{2 is obtained} as a white foam which has the Rf 0.36 in chloroform-methanol 9: 1.

8.68 g of dicyclohexylcarbodiimide and 5.86 g of N-hydroxybenzotriazole are added to 11.46 g of Z-Phe-OH and 12.64 g of H-Ala-Glu (OBu ^t ) ₂ in 50 ml of dimethylformamide at 0 °. After 15 hours at 4 °, the mixture is filtered off, the filtrate is evaporated and the residue is recrystallized from ethyl acetate-hexane. The Z-Phe-Ala-Glu (OBu ^t ) ₂ of mp. 161-163 ° is obtained. Rf (toluene-acetone 1: 1) = 0.63.

6.4 g of this product are dissolved in 50 ml of methanol and, after addition of 0.5 g of Pd carbon (10%), hydrogenated at 20 ° for 20 minutes. The catalyst is filtered off and the filtrate is evaporated, the H-Phe-Ala-Glu (OBu ^t ) _{2 being obtained} as a white foam. Rf (chloroform-methanol 7: 3) = 0.75.

To 4.43 g of Z-Ser (Bu ^t ) -OH and 3.18 g of HCl.H-Ser (Bu ^t ) -OMe in 30 ml of dimethylformamide, 1.89 ml of N-ethylmorpholine, 2.29 are added at 0 ° g HOBt and 3.4 g dicyclohexylcarbodiimide. After 2 hours at 0 ° and 15 hours at 20 °, the mixture is filtered off, the filtrate is evaporated and taken up in ethyl acetate. After extraction of the solution with sodium bicarbonate, dilute hydrochloric acid and water, the mixture is evaporated to give the Z-Ser (Bu ^t ) -Ser (Bu ^t ) -CMe as an oil. Rf (toluene-acetone 1: 1) = 0.70.

7.5 ml of hydrazine hydrate are added to a solution of 6.8 g of this product in 40 ml of methanol and, after 24 hours at 20 °, evaporated to about half the volume. It is taken up in 250 ml of ethyl acetate and extracted with water. The ethyl acetate solution is concentrated and the Z-Ser (Bu ^t ) -Ser (Bu ^t ) -NH ― NH ₂ obtained is precipitated with petroleum ether. Rf (toluene-acetone 1: 1) = 0.50.

2.26 g of this product are dissolved in 20 ml of dimethylformamide and tert at -10 ° with 7.022 ml of 1.78 N-HCl in ethyl acetate and 0.635 ml. Butyl nitrite added. After 10 minutes at -10 °, a solution of 2.38 g of H-Phe-Ala-Glu (OBu ^t ) ₂ and 2.52 ml of N-ethylmorpholine is added dropwise and the mixture is kept at -10 ° for one hour and at 0 ° for 15 hours . The solution is then evaporated, the residue taken up in ethyl acetate and washed with sodium bicarbonate, dilute hydrochloric acid and water. The Z-Ser (Bu ^t ) -Ser (Bu ^t ) -Phe-Ala-Glu (OBu ^t ) ₂ obtained after evaporation of the solution is recrystallized from ethyl acetate-hexane. Mp 213-215 °, Rf (toluene-acetone 1: 1) = 0.70.

2.30 g of this product are dissolved in 40 ml of methanol and, after addition of 0.5 g of Pd carbon (10%), hydrogenated at 20 ° for 2 hours. The catalyst is filtered off and the filtrate is evaporated, the H-Ser (Bu ^t ) -Ser (Bu ^t ) -Phe-Ala-Glu (OBu ^t ) _{2 being obtained} as a white foam. Rf (toluene-acetone 1: 1) = 0.23.

• 6,3 g (2.78 mMol) N-Palmitoyl-S-[2(R),3-dipalmitoyloxypropyl]-(R)-cystein-benzhydrylester löst man in einer Mischung aus 12 ml Trifluoressigsäure und 48 ml Methylenchlorid. Nach einer Stunde bei Raumtemperatur dampft man die Lösung ein und verreibt den öligen Rückstand mit Wasser. Das auskristallisierte N-Palmitoyl-S-[2(R),3-dipalmitoyloxypropyl]-(R)-cystein wird abgesaugt, getrocknet und zur Entfernung des Benzhydrols dreimal mit Benzin verrieben. Man kristallisiert den Rückstand aus Aceton-Wasser und erhält so farblose Kristalle dieses Produkts vom Smp. 71-75° und [α]²⁰ _D =― 1 ° (Dioxan).

The N-palmitoyl-S- [2 (R), 3-dipalmitoyloxypropyl) - (R) -cysteine used as the starting material and the diastereomeric mixture of this compound with the corresponding 2 (S) compound, which is abbreviated referred to here as 2 (R, S) can be made as follows:

• 6.3 g (2.78 mmol) of N-palmitoyl-S- [2 (R), 3-dipalmitoyloxypropyl] - (R) -cysteine-benzhydryl ester are dissolved in a mixture of 12 ml of trifluoroacetic acid and 48 ml of methylene chloride. After one hour at room temperature, the solution is evaporated and the oily residue is triturated with water. The crystallized N-palmitoyl-S- [2 (R), 3-dipalmitoyloxypropyl] - (R) -cysteine is filtered off, dried and triturated three times with gasoline to remove the benzhydrol. The residue is crystallized from acetone-water and thus colorless crystals of this product of mp 71-75 ° and [α] ²⁰ _D = - 1 ° (dioxane) are obtained.

• 12 g (20 mMol) N-Palmitoyl-S-[2(R),3-dihydroxypropyl]-(R)-cystein-benzhydrylester löst man in einer Mischung aus 60 ml Pyridin und 60 ml Methylenchlorid und tropft bei 0-10° 14,5 ml (13,2 g) Palimitinsäurechlorid zu. Nach 2 Stunden bei Raumtemperatur ist die Reaktion gemäss Dünnschichtchromatogramm beendet. Man gibt zur Suspension (Pyridinhydrochlorid) 10 ml Methanol, rührt 20 Minuten bei 25° und dampft dann das Reaktionsgemisch weitgehend ein. Man nimmt den Rückstand in Chloroform auf, schüttelt je zweimal mit 0,5 N-Salsäure, 10% Natriumbicarbonat und Wasser aus, trocknet die Chloroform-Phase mit Natriumsulfat und dampft sie zum Syrup ein. Den reinen Ester erhält man durch Chromatographie dieses Syrups über Kieselgel in Chloroform. Die chromatographisch einheitlichen Fraktionen (DC-Kontrolle auf Kieselgel in Chloroform: Essigester 98:2, Rf=0,5) werden eingedampft und aus Aceton umkristallisiert. Man erhält farblose Kristalle des N-Palmitoyl-S-[2(R),3-dipalmitoyloxypropyl]-(R)-cystein-benzhydrylesters vom Smp. 69-71 °.

The above ester is obtained in the following way:

• 12 g (20 mmol) of N-palmitoyl-S- [2 (R), 3-dihydroxypropyl] - (R) -cysteine-benzhydryl ester are dissolved in a mixture of 60 ml of pyridine and 60 ml of methylene chloride and added dropwise at 0-10 ° 14.5 ml (13.2 g) of palimitic acid chloride. After 2 hours at room temperature, the reaction according to the thin layer chromatogram is complete. 10 ml of methanol are added to the suspension (pyridine hydrochloride), the mixture is stirred at 25 ° for 20 minutes and then the reaction mixture is largely evaporated. The residue is taken up in chloroform, shaken twice with 0.5 N-salic acid, 10% sodium bicarbonate and water, the chloroform phase is dried with sodium sulfate and evaporated to give the syrup. The pure ester is obtained by chromatography of this syrup over silica gel in chloroform. The chromatographically uniform fractions (TLC control on silica gel in chloroform: ethyl acetate 98: 2, Rf = 0.5) are evaporated and recrystallized from acetone. Colorless crystals of N-palmitoyl-S- [2 (R), 3-dipalmitoyloxypropyl] - (R) -cysteine-benzhydryl ester of mp 69-71 ° are obtained.

• 23,4 g (54 mMol) N-Palmitoyl-S-[2(R),3-dihydroxypropyl]-(R)-cystein löst man in 150 ml Chloroform-Aethanol-Gemisch 1:1 und tropft dazu bei Raumtemperatur eine Lösung von 13,6 g (70 mMol) Diphenyldiazomethan in 50 ml Chloroform. Nach 3 Stunden bei Raumtemperatur ist die anfänglich rote Lösung entfärbt und nach DC (Chloroform: Methanol = 9:1, Kieselgel) die Reaktion beendet.

The benzydrylester of N-palmitoyl-S- [2 (R), 3-dihydroxypropyl] - (R) -cysteine used above can be obtained, for example, in the following way:

• 23.4 g (54 mmol) of N-palmitoyl-S- [2 (R), 3-dihydroxypropyl] - (R) -cysteine are dissolved in 150 ml of chloroform-ethanol mixture 1: 1 and a solution is added dropwise at room temperature of 13.6 g (70 mmol) of diphenyldiazomethane in 50 ml of chloroform. After 3 hours at room temperature, the initially red solution is decolorized and the reaction is ended after TLC (chloroform: methanol = 9: 1, silica gel).

It is evaporated to the dry state in a vacuum and the residue is extracted cold with petroleum ether. The residue is filtered through silica gel with methylene chloride as the solvent. This gives colorless crystals of the benzhydryl ester of mp. 88-91 °.

• 50 g (139 mMol) N-Palmitoyl-(R)-cystein, 12,4 g Glycid und 45 g Kaliumcarbonat werden in 375 ml Aethanol unter Stickstoff und gutem Rühren 16 Stunden auf 80° erhitzt. Nach dem Abkühlen auf Raumtemperatur säuert man mit 2 N-Salzsäure auf pH ca. 4 an und versetzt mit Wasser; dabei fällt das Produkt aus. Man saugt ab und wäscht mit Wasser bis das Filtrat frei von Chlorionen ist. Nach dem Trocknen des Nutschgutes im Vakuum kristallisiert man aus Essigester um. Man erhält farblose Kristalle des Diastereomerengemisches des N-Palmitoyl-S-[2(R),3-dihydroxypropyl]-(R)-cysteins und N-Palmitoyl-S-[2(S),3-dihydroxypropyl]-(R)-cysteins vom Smp. 76-155°.

The procedure for the preparation of the N-palmitoyl-2- [2 (R, S), 3-dihydroxypropyl] - (R) -cysteine is as follows:

• 50 g (139 mmol) of N-palmitoyl- (R) -cysteine, 12.4 g of glycide and 45 g of potassium carbonate are heated in 375 ml of ethanol under nitrogen and with good stirring for 16 hours at 80 °. After cooling to room temperature, the mixture is acidified to pH about 4 with 2 N hydrochloric acid and water is added; the product fails. It is suctioned off and washed with water until the filtrate is free of chlorine ions. After drying the suction filter in vacuo, the product is recrystallized from ethyl acetate. Colorless crystals of the diastereomer mixture of N-palmitoyl-S- [2 (R), 3-dihydroxypropyl] - (R) -cysteine and N-palmitoyl-S- [2 (S), 3-dihydroxypropyl] - (R) are obtained. -cysteins of mp 76-155 °.

The N is obtained by reacting the N-palmitoyl-2- [2 (R, S), 3-dihydroxypropyl] - (R) -cysteine with diphenyldiazomethane, as indicated above for the R compound S, and subsequent palmitoylation, as also stated above -Palmitoyl-S- [2 (R, S), 3-dipalmitoyloxypropyl] - (R) -cysteine-benzhydryl ester: colorless crystals of mp 84-85 °.

If this mixture of diastereomers (19.8 g) is chromatographed on a column with 250 g of silica gel (Merck) with methylene chloride as solvent, the first fraction (1.55 g) gives the R-diastereomer (Rf = 0, 61, methylene chloride: ethyl acetate = 98: 2, thin layer, silica gel), in a second fraction 12.7 g of an R, S-diastereomer mixture and in a third fraction (1.1 g) of the S-diastereomer (Rf = 0.53 in methylene chloride: ethyl acetate = 98: 2).

The (S) -diastereomer is used to obtain N-palmitoyl-S- [2 (S), 3-dipalmitoyloxypropyl] (R) -cysteine in the manner described above; colorless crystals, mp. 64-65 ° [α] ²⁰ _D = + 1 ° (dioxane).

• N-Myristoyl-S-[2(R),3-dihydroxypropyl]-(R)-cystein, farblose Kristalle vom Smp. 140-150°, und das
• N-Stearoyl-S-[2(R),3-dihydroxypropyl]-(R)-cystein, farblose Kristalle vom Smp. 140-150°;
• N-Lauroyl-S-[2(R),3-dihydroxypropyl]-(R)-cystein, farblose Kristalle vom Smp. 140-150°.

The new lipopeptides to be prepared in accordance with the invention for the preparation of the new lipopeptides mentioned in this example, such as N-palmitoyl-S- [2 (R), 3-dipalmitoyloxypropyl] -Cys-Ser-Ser-Phe-Ala-Glu-OH required starting materials are, for example

• N-myristoyl-S- [2 (R), 3-dihydroxypropyl] - (R) -cysteine, colorless crystals of mp. 140-150 °, and that
• N-stearoyl-S- [2 (R), 3-dihydroxypropyl] - (R) -cysteine, colorless crystals of mp 140-150 °;
• N-Lauroyl-S- [2 (R), 3-dihydroxypropyl] - (R) -cysteine, colorless crystals of mp 140-150 °.

They can be produced according to the information in example 4.

• N-Myristoyl-S-[2(R),3-dihydroxypropyl]-(R)-cystein-benzhydrylester, farblose Kristalle vom Smp. 90-92°;
• N-Stearoyl-S-[2(R),3-dihydroxypropyl]-(R)-cystein-benzhydrylester, farblose Kristalle vom Smp. 95°-97°;
• N-Lauroyl-S-[2(R),3-dihydroxypropyl]-(R)-cystein-benzhydrylester, farblose Kristalle vom Smp. 95-97°.

The benzhydryl esters to be produced according to the above information have the following data:

• N-myristoyl-S- [2 (R), 3-dihydroxypropyl] - (R) -cysteine-benzhydryl ester, colorless crystals of mp 90-92 °;
• N-stearoyl-S- [2 (R), 3-dihydroxypropyl] - (R) -cysteine-benzhydryl ester, colorless crystals of mp 95 ° -97 °;
• N-Lauroyl-S- [2 (R), 3-dihydroxypropyl] - (R) -cysteine-benzhydryl ester, colorless crystals of mp 95-97 °.

• N-Lauroyl-S-[2(R),3-dipalmitoyloxypropyll-(R)-cystein-benzhydrylester, Smp, 65-68°;
• N-Myristoyl-S-[2(R),3-dipalmitoyloxypropyl]-(R)-cystein-benzhydrylester, Smp. 68-70°;
• N-Stearoyl-S-[2(R),3-dipalmitoyloxypropyl]-(R)-cystein-benzhydrylester, Smp. 70-72°.

The following N-acyl-S- [2 (R), 3-diacyloxypropyl] - (R) -cysteine-benzhydryl ester with different acyl groups on the N and on the O atoms can be obtained in an analogous manner to that described:

• N-Lauroyl-S- [2 (R), 3-dipalmitoyloxypropyll- (R) -cysteine-benzhydryl ester, m.p. 65-68 °;
• N-myristoyl-S- [2 (R), 3-dipalmitoyloxypropyl] - (R) -cysteine-benzhydryl ester, mp 68-70 °;
• N-stearoyl-S- [2 (R), 3-dipalmitoyloxypropyl] - (R) -cysteine-benzhydryl ester, mp 70-72 °.

• N-Lauryl-S-[2(R),3-dipalmitoyloxypropyl]-(R)-cystein, farblose Kristalle, Smp. 70-72°;
• N-Myristoyl-S-[2(R),3-dipalmitoyloxypropyl]-(R)-cystein, farblose Kristalle, Smp. 71-73°;
• N-Stearoyl-S-[2(R),3-dipalmitoyloxypropyl]-(R)-cystein, farblose Kristalle, Smp. 74-77°.

Other compounds to be used as an intermediate, which can be prepared according to the above information, are:

• N-Lauryl-S- [2 (R), 3-dipalmitoyloxypropyl] - (R) -cysteine, colorless crystals, mp 70-72 °;
• N-myristoyl-S- [2 (R), 3-dipalmitoyloxypropyl] - (R) -cysteine, colorless crystals, mp 71-73 °;
• N-Stearoyl-S- [2 (R), 3-dipalmitoyloxypropyl] - (R) -cysteine, colorless crystals, mp 74-77 °.

Example 6

• N-Palmitoyl-S-[2(R),3-dilauroyloxypropyl]-(R)-cystein-benzhydrylester, farbloses Wachs;
• N-Palmitoyl-S-[2(R),3-distearoyloxypropyl]-(R)-cystein-benzhydrylester, farblose Kristalle, Smp. 80―82°;
• N-Palmitoyl-S-[2(R),3-dioleoyloxypropyl]-(R)-cystein-benzhydrylester, farbloses Oel;
• N-Palmitoyl-S-[2(R),3-dibehenoyloxypropyl]-(R)-cystein-benzhydrylester, farblose Kristalle, Smp. 85-88°;
• N-Palmitoyl-S-[2(R),3-di-(dihydrosterkuloyloxypropyl]-(R)-cystein-benzhydrylester, farbloses Wachs.

The following starting materials for the production of lipopeptides according to the application are prepared by means of the reactions described in Example 5:

• N-palmitoyl-S- [2 (R), 3-dilauroyloxypropyl] - (R) -cysteine-benzhydryl ester, colorless wax;
• N-palmitoyl-S- [2 (R), 3-distearoyloxypropyl] - (R) -cysteine-benzhydryl ester, colorless crystals, mp. 80-82 °;
• N-palmitoyl-S- [2 (R), 3-dioleoyloxypropyl] - (R) -cysteine-benzhydryl ester, colorless oil;
• N-palmitoyl-S- [2 (R), 3-dibehenoyloxypropyl] - (R) -cysteine-benzhydryl ester, colorless crystals, mp 85-88 °;
• N-Palmitoyl-S- [2 (R), 3-di- (dihydrosterkuloyloxypropyl] - (R) -cysteine-benzhydryl ester, colorless wax.

• N-Palmitoyl-S-[2(R),3-dilauroyloxypropyl]-(R)-cystein, farbloses Oel, Rf=0,27;
• N-Palmitoyl-S-[2(R),3-distearoyloxypropyl]-(R)-cystein, farblose Kristalle, Smp. 82-85°;
• N-Palmitoyl-S-[2(R),3-dioleoyloxypropyl]-(R)-cystein, farbloses Oel, Rf in Chloroform: Methanol = 9:1 (Dünnschicht- Kieselgel-Merck) = 0,35;
• N-Palmitoyl-S-[2(R),3-dibehenoyloxypropyl]-(R)-cystein, farblose Kristalle., Smp. 87-90°;
• N-Palmitoyl-S-[2(R),3-di-(dihydrosterkuloyloxypropyl]-(R)-cystein, farbloses Wachs, Rf=0,38 (gleiche Bedingungen wie oben für das N-Palmitoyl-2,3-dioleoyloxy-Derivat angegeben).

The corresponding non-esterified substituted cysteines have the following data:

• N-palmitoyl-S- [2 (R), 3-dilauroyloxypropyl] - (R) -cysteine, colorless oil, Rf = 0.27;
• N-palmitoyl-S- [2 (R), 3-distearoyloxypropyl] - (R) -cysteine, colorless crystals, mp 82-85 °;
• N-Palmitoyl-S- [2 (R), 3-dioleoyloxypropyl] - (R) -cysteine, colorless oil, Rf in chloroform: methanol = 9: 1 (thin layer silica gel Merck) = 0.35;
• N-palmitoyl-S- [2 (R), 3-dibehenoyloxypropyl] - (R) -cysteine, colorless crystals. Mp 87-90 °;
• N-Palmitoyl-S- [2 (R), 3-di- (dihydrosterkuloyloxypropyl] - (R) -cysteine, colorless wax, Rf = 0.38 (same conditions as above for the N-palmitoyl-2,3-dioleoyloxy -Derivative specified).

• H-Phe-Phe-Asn-Ala-Lys-OH
• H-Glu-Gln-Asn-Ala-Lys-OH
• H-Ser-Ser-Asn-Ala-Glu-OH

These cysteine derivatives are condensed with the following peptides as described in Example 5:

• H-Phe-Phe-Asn-Ala-Lys-OH
• H-Glu-Gln-Asn-Ala-Lys-OH
• H-Ser-Ser-Asn-Ala-Glu-OH

The corresponding lipopeptides according to the invention are thus obtained.

Example 7

0.9 g of N-palmitoyl-S-2 (R), 3 (R), 4-tripalmitoyloxy-butyl- (R) -cysteine-benzhydryl ester in a mixture of 3 ml of trifluoroacetic acid and 12 ml of methylene chloride are left for 2 hours at room temperature stand. Then it is evaporated to dryness, the residue is triturated with ice water, suction filtered and the solid is dried. Then it is extracted with gasoline and recrystallized from acetone. Colorless crystals of N-palmitoyl-S-2 (R), 3 (R), 4-tripalmitoyloxy-butyl- (R) -cysteine with a melting point of 76-76.5 ° and a specific rotation [α] ²⁰ _{D are obtained} = ―5 ° (c = 0.819, dioxane).

• 1 g N-Palmitoyl-S-2(R),3(R),4-trihydroxybutyl-(R)-cystein-benzhydrylester werden in einer Mischung aus 6 ml Pyridin in 5 ml Methylenchlorid mit 1,83 ml Palmitinsäurechlorid acyliert. Nach 18 Stunden bei Raumtemperatur gibt man 1 ml Methanol zu und dampft zur Trockne. Den Rückstand filtriert man über 100 g Kieselgel Merck mit Chloroform als Lösungsmittel. Die nach Dünnschichtchromatographie (CHCI₃: Essigester = 98:2) reinen Fraktionen geben nach Eindampfen farblose Kristalle vom Smp. 60-62°.

The benzhydryl ester used as the starting material is obtained in the following way:

• 1 g of N-palmitoyl-S-2 (R), 3 (R), 4-trihydroxybutyl- (R) -cysteine-benzhydryl ester are acylated in a mixture of 6 ml of pyridine in 5 ml of methylene chloride with 1.83 ml of palmitic acid chloride. After 18 hours at room temperature, 1 ml of methanol is added and the mixture is evaporated to dryness. The residue is filtered through 100 g of silica gel Merck with chloroform as a solvent. The fractions which are pure after thin layer chromatography (CHCI ₃ : ethyl acetate = 98: 2) give colorless crystals of mp 60-62 ° after evaporation.

• 2,53 g N-Palmitoyl-S-2(R),3(R),4-trihydroxybutyl-(R)-cystein lässt man in einer Mischung aus 12 ml Aethanol und 32 ml Chloroform mit 1,37 g Diphenyldiazomethan 15 Stunden bei Raumtemperatur reagieren. Man dampft zur Trockne und reinigt den entstandenen Benzhydrylester an 150 g Kieselgel in Chloroform als Lösungsmittel. Rf = 0,3 (CHC1₃: Aceton = 95:5) Kieselgel Merck, Dünnschicht.

The starting material for the above reaction is obtained as follows:

• 2.53 g of N-palmitoyl-S-2 (R), 3 (R), 4-trihydroxybutyl- (R) -cysteine are left in a mixture of 12 ml of ethanol and 32 ml of chloroform with 1.37 g of diphenyldiazomethane for 15 hours react at room temperature. The mixture is evaporated to dryness and the benzhydryl ester formed is purified on 150 g of silica gel in chloroform as solvent. Rf = 0.3 (CHC1 ₃ : acetone = 95: 5) silica gel Merck, thin layer.

N-Palmitoyl-S-2 (R), 3 (R), 4-trihydroxybutyl- (R) -cysteine is obtained in the following way.

20 g of N-palmitoyl cysteine, 27.6 g of 1-toxyl-2,4-ethylidene-D-erythritol and 17 g of potash stirred under nitrogen in 240 ml of ethanol at 80 ° for 15 hours. Then the salts are filtered off and the filtrate is evaporated to dryness. The residue is taken up in 200 ml of H ₂ 0: tetrahydrofuran = 1: 1 and acidified to pH = 3 with 2 N hydrochloric acid. Then shake out three times with ether, adjust the pH of the ether phase to 4.5 with pyridine, shake out with water, dry the ether phase and evaporate to dryness. This residue is purified over 200 g of Merck silica gel with chloroform: acetone = 8: 2 (1 1) and then with CHCl ₃ : MeOH = 6: 4 (1.2 1) as the eluent.

The pure fractions are evaporated, triturated with hexane and recrystallized from ethyl acetate-petroleum ether.

Colorless crystals of N-palmitoyl-S - [(2,4-ethylidene) erythrityl] - (R) -cysteine of mp 6265 ° are obtained.

The hydrolysis of this compound is carried out with 3 percent. HBF _{4 carried out} at 80 ° for 4.5 hours.

8.2 g of ethylidene derivative in 100 ml of dimethoxyethane and 100 ml of 3 percent. HBF treated. Then it is cooled to 0 °, whereby the hydrolysis product fails. After suction and drying, one recrystallizes from ethyl acetate. Mp 160-165 °, sintering from 93 °. Rf = 0.285 (CHCl ₃ : MeOH = 6: 4) thin layer, silica gel, Merck.

Example 8

• N-Stearoyl-S-[2(R),3-dipalmitoyloxypropyl]-Cys-Ser-Ser-Asn-Ala-Glu-Ile-Asp-Glu-OH, Rf (157) = 0,32;
• N-Palmitoyl-S-[2(R),3(R),4-tripalmitoyloxybutyl]-Cys-Ser-Ser-Asn-Ala-Lys-OH, Rf (157) = 0,55;
• N-Palmitoyl-S-[2(R),3(R),4-tripalmitoyloxybutyl]-Cys-Ser-Ser Asn-Ala-Lys-Ile-Asp-Glu-OH, Rf (157) = 0,38.

Analogously to the information in Example 5, N-stearoyl-S- [2 (R), 3-dipalmitoyloxypropylJ- (R) -cysteine and N-palmitoyl-S- [2 (R), 3 (R), 4- tripalmitoyloxybutyl] - (R) -cysteine produced the following lipopeptides:

• N-stearoyl-S- [2 (R), 3-dipalmitoyloxypropyl] -Cys-Ser-Ser-Asn-Ala-Glu-Ile-Asp-Glu-OH, Rf (157) = 0.32;
• N-Palmitoyl-S- [2 (R), 3 (R), 4-tripalmitoyloxybutyl] -Cys-Ser-Ser-Asn-Ala-Lys-OH, Rf (157) = 0.55;
• N-Palmitoyl-S- [2 (R), 3 (R), 4-tripalmitoyloxybutyl] -Cys-Ser-Ser Asn-Ala-Lys-Ile-Asp-Glu-OH, Rf (157) = 0.38.

Example 9

• PC(R,S)-Ser-Ser-Asn-OH
• PC(R)-Ser-Ser-Asn-Ala-Lys-OH
• PC(R,S)-Ser-Ser-Asn-Ala-Lys-OH
• PC(R)-Ser-OH
• PC(R,S)-Ser-OH
• PC(R,S)-Ser-Ser-Asn-Ala-OH
• PC(R)-Phe-Ile-Ile-Phe-Ala-OH
• PC(R,S)-Phe-Ile-Ile-Phe-Ala-OH
• PC(R)-Val-Lys-Val-Tyr-Pro-OH
• PC(R,S)-Val-Lys-Val-Tyr-Pro-OH
• PC(R)-Ala-Ile-Gly-Val-Gly-Ala-Pro-NH₂
• PC(R,S)-Ala-Ile-Gly-Val-Gly-Ala-Pro-NH₂
• PC(R)-Ser-Ser-Asn-Ala-Lys-Ile-Asp-Glu-OH
• PC(R,S)-Ser-Ser-Asn-Ala-Lys-Ile-Asp-Glu-OH

Analogously to the information in Example 5, the lipopeptides listed below are prepared from N-palmitoyl-2 (R), 3-dipalmitoyloxypropyl- (R) -cysteine or N-palmitoyl 2 (R, S), 3-dipalmitoyloxypropyl-cysteine, the Residues of the cysteine derivatives just mentioned are referred to as PC (R) or PC (R, S):

• PC (R, S) -Ser-Ser-Asn-OH
• PC (R) -Ser-Ser-Asn-Ala-Lys-OH
• PC (R, S) -Ser-Ser-Asn-Ala-Lys-OH
• PC (R) -Ser-OH
• PC (R, S) -Ser-OH
• PC (R, S) -Ser-Ser-Asn-Ala-OH
• PC (R) -Phe-Ile-Ile-Phe-Ala-OH
• PC (R, S) -Phe-Ile-Ile-Phe-Ala-OH
• PC (R) -Val-Lys-Val-Tyr-Pro-OH
• PC (R, S) -Val-Lys-Val-Tyr-Pro-OH
• PC (R) -Ala-Ile-Gly-Val-Gly-Ala-Pro-NH ₂
• PC (R, S) -Ala-Ile-Gly-Val-Gly-Ala-Pro-NH ₂
• PC (R) -Ser-Ser-Asn-Ala-Lys-Ile-Asp-Glu-OH
• PC (R, S) -Ser-Ser-Asn-Ala-Lys-Ile-Asp-Glu-OH

Furthermore, the R, S-epimer mixtures of all lipopeptides described in the previous examples are prepared.

Claims (33)

1. Compounds of the formula

wherein R, and R₂ each represent a saturated or unsaturated aliphatic or mixed aliphatic-cycloaliphatic hydrocarbon radical which has 11-21 C atoms and which is also optionally substituted by oxygen functions, R₃ represents hydrogen or the radical R,-CO-O-CH2-, where R, has the same meaning, and X represents a natural aliphatic amino acid linked by a peptide bond and having a free, esterified or amidated carboxyl group, or an amino acid sequence of 2-10 natural aliphatic amino acids, the terminal carboxyl group of which is free or in the ester or amide form, with the asymmetric centres denoted by ^* and ^** possessing the absolute R- and S- or R-configurations, respectively; and, possibly, mixtures of the R- and S-compounds epimeric on the C^** atoms, and salts and complexes of all these compounds.

2. Compounds according to Claim 1, wherein the acyl groups R,CO and R₂CO are derived from saturated or unsaturated fatty acids having 14-18 C atoms, and wherein R, and R₂ can be identical or different.

3. Compounds according to Claim 1, wherein the acyl groups are derived from palmitic, stearic, oleic, lauric, myristic, behenic, dihydrosterculic, malvalic, hydnocarpic or chaulmoogric acid.

4. Compounds according to any one of Claims 1-3, wherein amino acids carrying hydrophilic groups in X are the ionic amino acids: aspartic acid, glutamic acid, and/or oxyglutamic acid, or: lysine, ornithine, arginine and/or histidine, or asparagine, glutamine, serine and/or threonine, and amino acids carrying no hydrophilic groups in X are glycine, alanine, valine, norvaline, leucine, isoleucine and/or methionine.

5. Compounds according to any one of Claims 1-4, wherein in the peptide chain X the amino acids having hydrophilic character are linked directly with each other.

6. Compounds according to Claim 5, wherein the sequence of the amino acids having hydrophilic character is bound directly to the carboxyl group of the cysteine in the triacyl-glycerylcysteine part or tetraacyl-erythritylcysteine part.

7. Compounds according to any one of Claims 1-6, wherein X in the formula I represents an amino acid, or a peptide chain having 2-5 amino acids.

8. Compounds according to Claim 1 of the formula

their salts and complexes, wherein X represents a natural aliphatic amino acid linked by a peptide bond and having a hydrophilic group and a free, esterified or amidated carboxyl group, or an amino acid sequence of 2-5 natural aliphatic amino acids of which half carry at least one hydrophilic group, and of which the terminal carboxyl group is free or in the ester or amide form, with the asymmetic centres denoted by ^* and ^** possessing the R-configuration.

9. Compounds according to any one of Claims 1-6, wherein X in formula I of Claim 1 represents an unnatural amino acid sequence compared with that of the known murein-lipoprotein-degradation lipopeptides.

10. Compounds corresponding to those of Claim 8, in which however the configuration on the C^** atom is S instead of R, and the mixtures of the R- and S-diastereomers.

11. Compounds according to any one of Claims 1-6 of the formula

wherein R, and R₂ have the meanings given under the formula I, and X represents a peptide chain having 6-10 amino acids, with the asymmetric centres denoted by ^* and ^** possessing the R-configuration, and also the compounds having on the C^** atom the (S)-configuration, as well as R- and S-diastereomers mixtures, and salts and complexes thereof.

12. Compounds according to any one of Claims 1-6 of the formula

wherein R_i and R₂ have the meanings given in any one of Claims 1-6, and X represents an amino acid, or a peptide chain having 2-5 amino acids, and mixtures of the compounds stereoisomeric on the C^** atoms, and salts and complexes thereof.

13. The amides and esters of the compounds according to Claims 7-12, wherein an ester radical in the terminally esterified carboxyl group of X is derived from a lower aliphatic alcohol having 1-7 C atoms, an ethylene glycol or glycerin, and a terminal amide group in X is the amide group CONH₂, or an amide group derived from a lower aliphatic amine having 1-7 C atoms, or from pyrrolidine, piperidine or piperazine.

14. The lipopeptide: N-palmitoyl-S-[2(R),3-dipalmitoyloxypropyl]-Cys-Ser-Ser-Asn-Ala-Lys-OH, according to Claim 1.

15. The lipopeptide: N-palmitoyl-S-[2(R),3-dipalmitoyloxypropyl]-Cys-Ser-Ser-Phe-Ala-Glu-OH, according to Claim 1.

16. N - palmitoyl - S - [2(R),3 - di - (dihydrosterculoyloxypropyl] - Cys - Ser - Ser - Asn - Ala - Glu - OH, according to Claim 1.

17. The lipopeptide: N-lauroyl-S-[2(R),3-dipalmitoyloxypropylJ-Cys-Ser-Ser-Asn-Ala-Glu-OH, according to Claim 1.

18. N-Myristoyl-S-[2-(R),3-dipalmitoyloxypropyl]-Cys-Glu-Gln-Asn-Ala-Lys-OH, according to Claim 1.

19. N-Palmitoyl-S-[2(R,S),3-dilauroyloxypropyl]-Cys-Phe-Phe-Asn-Ala-Lys-OH, according to Claim 1.

20. N-palmitoyl-S-[2(R),3-dilauroyloxy]-Cys-Ser-Ser-Asn-Ala-Glu-OH, according to Claim 1.

21. N-palmitoyl-S-[3(R),3-dipalmitoyloxypropyl]-Cys-Ala-Ile-Gly-Val-Gly-Ala-Pro-NH₂, according to Claim 1.

22. N-palmitoyl-S-[2(R),3-dioleoyloxypropyl]-Cys-Ser-Ser-Asn-Ala-Glu-OH, according to Claim 1.

23. N-Palmitoyl-S-[2(R,S),3-dipalmitoyloxypropyl]-Cys-Phe-Phe-Asn-Ala-Lys-OH, according to Claim 1.

24. N - Stearoyl - S - [2(R),3 - dipalmitoyloxypropyl] - Cys - Ser - Ser - Asn - Ala - Glu - lie - Asp - Glu - OH, according to Claim 1.

25. The lipopeptides corresponding to the compounds of Claims 14-15, in which instead of palmitoyl radicals being present in the diacyloxypropyl radical, there are present lauroyl, stearoyl, oleoyl, behenoyl or dihydrosterculoyl radicals.

26. A compound of Claims 14-18, 21 and 23-24 having the R-configuration on the 2-C atom of the propyl group, a corresponding epimeric compound having the S-configuration, or R- and S-diastereomers mixtures.

27. Ammonium salts, alkali metal or alkaline-earth metal salts of acid compounds, and pharmaceutically applicable nontoxic acid addition salts of basic compounds, and copper-, zinc-, iron-or cobalt-complex salts of compounds according to any one of Claims 1-26.

28. Compound of the formula

wherein R₁ and R₂ each represent a saturated or unsaturated, aliphatic or mixed aliphatic-cycloaliphatic hydrocarbon radical which has 11-21 C atoms and which is also optionally substituted by oxygen functions, and R₃ represents hydrogen or the radical R₁―CO―OCH₂―, where R, has the same meaning, with the asymmetric centres denoted by ^* and ^** having the absolute R- and S- or R-configurations, respectively, and mixtures of the compounds stereoisomeric on the C^** atoms.

29. A compound according to Claim 28, wherein R₃ represents hydrogen, and wherein the acyl groups R_i and R₂ are derived from palmitic, stearic, oleic, lauric, myristic, behenic, dihydrosterculic, malvalic, hydnocarpic or chaulmoogric acid.

30. Pharmaceutical preparations containing a compound according to any one of Claims 1-28.

31. A compound according to any one of Claims 1-28 for application in a process for the therapeutic treatment of the animal or human body.

32. Compounds according to any one of Claims 1-28 for application as immunity-stimulating therapeutic preparations, especially as adjuvants in the production of antisera for therapy and diagnosis, and in the induction of immunologically activated lymphocyte populations for cell-transfer processes, or as prophylactics or therapeutics against infectious diseases in humans and animals.

33. A process for producing compounds of the formula

wherein R, and R₂ each represent a saturated or unsaturated aliphatic or mixed aliphatic-cycloaliphatic hydrocarbon radical which has 11-21 C atoms and which is also optionally substituted by oxygen functions, R₃ represents hydrogen or the radical R₁―CO―O―CH₂―, where R, has the same meaning, and X represents a natural aliphatic amino acid linked by a peptide bond and having a free, esterified or amidated carboxyl group, or an amino acid sequence of 2-10 natural aliphatic amino acids, the terminal carboxyl group of which is free or in the ester or amide form, with the asymmetric centres denoted by ^* and ^** possessing the absolute R- and S- or R-configurations, respectively; and optionally mixtures of the R- and S-compounds epimeric on the C^** atoms, and salts and complexes thereof, whereby

a) in a compound of the formula

wherein R₁, R₂ and R₃ have the meanings given under the formula (I) of Claim 1, and Y represents an amino acid or amino acid sequence corresponding to X in the said formula I, wherein however at least one of the hydrophilic groups substituting the amino acids and/or the terminal carboxyl group is (are) protected by a protective group which can be split off under neutral or mild acid conditions, or in a salt of such a compound, the protective groups(s) is or are split off; or

b) a compound of the formula

wherein R₁, R₂ and R₃ have the meanings given under the said formula (I), and W represents OH or an amino acid or an incomplete sequence of amino acids according to X in the said formula I, is reacted with a compound of the formula

where Z₁ represents a group corresponding to the group X in the said formula (I), or an amino acid sequence or amino acid complementary to the stated amino acid or incomplete sequence of amino acids according to X, in which sequences however no free amino groups are present, or with a salt thereof; or

c) a compound of the formula

wherein R₂ has the meaning given under the said formula (I), and R3 represents hydrogen or the group H―O―CH₂―, and Z₂ represents a group corresponding to X in the said formula (I), wherein however no free hydroxyl groups are present, or a salt of this compound, is acylated in a manner known per se; and, if required, in resulting compounds having a free terminal carboxyl group this group is converted into an amide group or ester group, and/or the compounds are converted into salts or complexes thereof.