Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
  • C07K7/04—Linear peptides containing only normal peptide links
  • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
  • C07K14/24—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
  • C07K14/245—Escherichia (G)
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/06—Dipeptides
  • C07K5/06008—Dipeptides with the first amino acid being neutral
  • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
  • C07K5/06026—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atom, i.e. Gly or Ala
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/06—Dipeptides
  • C07K5/06008—Dipeptides with the first amino acid being neutral
  • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
  • C07K5/0606—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing heteroatoms not provided for by C07K5/06086 - C07K5/06139, e.g. Ser, Met, Cys, Thr
  • C07K5/06069—Ser-amino acid
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/08—Tripeptides
  • C07K5/0802—Tripeptides with the first amino acid being neutral
  • C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
  • C07K5/081—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing O or S as heteroatoms, e.g. Cys, Ser
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/08—Tripeptides
  • C07K5/0802—Tripeptides with the first amino acid being neutral
  • C07K5/0812—Tripeptides with the first amino acid being neutral and aromatic or cycloaliphatic
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/10—Tetrapeptides
  • C07K5/1002—Tetrapeptides with the first amino acid being neutral
  • C07K5/1005—Tetrapeptides with the first amino acid being neutral and aliphatic
  • C07K5/1013—Tetrapeptides with the first amino acid being neutral and aliphatic the side chain containing O or S as heteroatoms, e.g. Cys, Ser
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides

Definitions

• the present invention relates to new lipopeptides and in particular to compounds of the formula wherein R 1 and R 2 are each 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 2nd where R 1 has the same meaning, and X is a peptide-bound natural aliphatic amino acid with 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, esterified ter or amidated form, mean, the asymmetry centers denoted by * or ** being the absolute S or.
• R and S compounds and optionally salts and complexes of these compounds, as well as processes for their synthetic production, and pharmaceutical preparations containing one or more of these lipopeptides together with a pharmaceutical carrier material and optionally together with other pharmaceutical compounds.
• Lipopeptides have already been described as degradation fragments of lipoproteins.
• Hantke and Braun (Eur. J. Biochem. 34, 284 - 296 (1973) were able to isolate lipopeptides or lipopeptide mixtures from the murein lipoprotein of the outer cell wall of Escherichia coli by means of enzymatic degradation following structure or corresponding formulas with a shortened polypeptide chain
• Y 1 , Y 2 ' Y 3 are acyl residues of various higher saturated and unsaturated fatty acids, such as those with 14-19 C atoms and others.
• the degradation lipopeptides obtained from murein protein contain the "glycerylcysteine" underlying.
• 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 (II), 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.
• R 1 and R 2 are saturated or unsaturated, aliphatic or aliphatic-cycloaliphatic, optionally also substituted by oxygen functions, hydrocarbon radicals with 11-21 C atoms, ie the acyl radicals are derived from saturated or unsaturated, aliphatic or cycloaliphatic carboxylic acids with 12-22, preferably with 14-18, carbon atoms, optionally oxygenated in the hydrocarbon radical.
• 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 ⁇ -elaostearic acid, stearolic acid, a-linolenic acid, and also among the cycloaliphatic-aliphatic, for example, are to be mentioned as such Dihydrosterkulkladre, Malvalic acid, Hydnocarpuskladre and Chaulmoograklare.
• Oxygenated acids of this type which are also suitable for the acyl radicals R 1 CO and R 2 CO, are, for example, those resulting from epoxidation of the abovementioned olefinic fatty acids and cycloaliphatic-aliphatic acids, for example ⁇ , l-epoxystearic acid, and also derivatives of these Acids mentioned above, which have, for example, one or more hydroxyl groups, such as ricinoleic acid.
• 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 formula (I) or. their salts consist of a maximum of 10 natural aliphatic amino acids, preferably at least half of which are hydrophilic Group, such as especially hydroxyl, amino, carboxyl, carbamide, guanidino or imidazolyl groups.
• hydrophilic Group such as especially hydroxyl, amino, carboxyl, carbamide, guanidino or imidazolyl groups.
• 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.
• Amino acids with a neutral character are particularly the amides, such as asparagine, glutamine and hyaroxy groups, primarily serine and threonine.
• 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.
• 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 among these in turn those in which this sequence of the hydrophilic amino acids to the carboxyl group of the cysteine in the Triacyl-Glycerylcystein or Tetraacyl-Erythritylcystein part is bound.
• lipopeptides are those in which the peptide chain X consists of a maximum of 5 amino acids.
• these are in particular those of the formula wherein R 1 , 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.
• 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.
• AcGCT acyl-glycerylcysteine part according to formula IV: then the corresponding types in which threonine, glutamine or asparagine replace the serine; also the connection types of the type: and the corresponding ones with threonine, glutamine or asparagine as exchange amino acids for serine, and the amides and carboxylic acid esters with terminal carbamide or ester groups, the compounds primarily being considered hen are in which the acyl radicals R 1 -CO, R 2 -CO in the acyl-glycerylcysteine part are the same and represent palmitoyl, stearoyl or oleoyl, and all those in which R 1 -CO and R 2 -CO are different and for example palmitoyl, stearoyl or oleoyl, and these residues occur in any combination and / or variation, such as those listed in the following table, the glyl radicals R 1 -CO, R 2 -CO in the
• 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 e.g. 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).
• a fifth group of lipopeptides according to the present invention is formed from those compounds according to formula (I) in which X denotes a peptide chain in which the sequence of the first 5 amino acids of those present in the known murein lipoprotein degradation lipopeptides is different, and in particular those in which R 3 is hydrogen, that is to say corresponding to the formula (IV) above, where R 1 -CO and R 2 -CO are the acyl radicals in the combinations mentioned which are preferably emphasized above or below, and their salts and complexes.
• Another subgroup consists of compounds according to formula (IV), in which X represents the “unnatural” sequence of amino acids compared to the murein-lipoprotein degradation products, and 6-10 Amino acids are present.
• X represents the “unnatural” sequence of amino acids compared to the murein-lipoprotein degradation products
• 6-10 Amino acids are present.
• 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: and their amides and esters, particularly those highlighted below.
• the terminal carboxyl group may be in amidated form.
• 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, such as pyrrolidine, piperidine or piperazine.
• Specific amides are, for example, the unsubstituted or the methyl, ethyl, dimethyl or diethyl amides of all the specific lipopeptides according to the present invention mentioned above or such amides of the compounds described in the illustrative examples.
• 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.
• 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.
• 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 are mentioned.
• the present new lipopeptides are neutral, acidic or basic compounds depending on the nature of their substituents. If excess acidic groups are present, they form salts with bases such as ammonium salts or salts with alkali or alkaline earth metals, e.g. Sodium, potassium, calcium or magnesium; however, if excess basic groups are present, they form acid addition salts.
• bases such as ammonium salts or salts with alkali or alkaline earth metals, e.g. Sodium, potassium, calcium or magnesium; however, if excess basic groups are present, they form acid addition salts.
• Acid addition salts are particularly pharmaceutically usable, non-toxic acid addition salts, such as those with inorganic acids, for example hydrochloric, hydrobromic, nitric, sulfuric or phosphoric acids, or with organic acids, such as organic carboxylic acids, for example 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, 2-acetoxybenzoic acid, embonic acid, nicotinic acid or isonicotinic acid, or organic sulfonic acids, for example methanesulfonic acid, ethanesulfonic acid, 2-hydroxy-ethanesulfonic acid, ethane-1,2-disulfonic acid, benzenesulf
• 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 (I) 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 pg / 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 with the most effective B cell mitogens known [dextran sulfate, E. coli lipopolysaccharide, PPD (purified protein derivative)] is achieved, with the new compounds of the present application even high concentrations having no lymphocytotoxic effect.
• the new compounds of the formula (I), their salts and complexes and mixtures are also able to induce the formation of antibody-producing cells in concentrations of 0.3-60 ⁇ g / ml in spleen cell cultures of normal mice (increase in the 19S-plaque-forming cells by a factor of 20 to 50 above the control value (in the absence of the stimulating substances):
• specific antibodies against sheep erythrocytes are formed without adding sheep erythrocytes to the cultures for immunization.
• 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).
• 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.
• the compounds mentioned in a concentration range of 10-100 ⁇ g / ml can significantly (up to 10 times) potentiate the reactivity of cortisone-resistant thymus cells with allogeneic irradiated stimulator lymphocytes.
• 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 immune potentiation).
• the new lipopeptides according to the present invention can thus be used as adjuvants in a mixture 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.
• 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.
• the new lipopeptides can also be used to promote immune reactions in humans and animals that are already taking place, even without simultaneous antigen supply.
• the connections are therefore suitable especially for the stimulation of the body's defense, e.g. with chronic and acute infections or with selective (antigen-specific) immunological defects, as well as with congenital, but also with acquired general (ie non-antigen-specific) immunological 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.
• the substances mentioned can thus preferably also be administered in combination with antibiotics, chemotherapeutic agents or other healing methods in order to counteract immunological damage.
• the substances described are also suitable for the general prophylaxis of infectious diseases in humans and animals.
• the new lipopeptides can be made according to methods known per se or their new methods described here. getting produced.
• the protective groups in the process according to variant a) are in particular those known from the synthesis of peptides.
• Protective groups for amino groups are, for example, acyl or aralkyl groups such as formyl, trifluoroacetyl, phthaloyl, benzenesulfonyl, p-toluenesulfonyl, o-nitrophenylsulfenyl, 2,4-dinitrophenylsulfenyl groups (these sulfenyl groups can also be reacted with nucleophilic reagents, e.g. sulfophilic reagents, e.g.
• Thiosulfates optionally substituted, such as by lower alkoxy groups, especially o- or p-methoxy groups
• benzyl, or diphenyl or triphenylmethyl 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.
• azo groups substituted arylmethyloxycarbonyl 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, for example carbobenzoxy, p-bromo or p-chlorocarbobenzoxy, p-nitrocarbobenzoxy or p-methoxycarbobenzoxy , p-Phenylazo-benzyloxycarbonyl and p- (p'-methoxy-phenylazo) -benzyloxycarbonyl, 2-tolyl-isopropyloxycarbonyl and in particular 2- (p-biphenylyl) -isopropyloxycarbonyl, as well as aliphatic oxycarbonyl groups such as adamantyloxycarbonyl, cyclopentyloxycarbonyl, trichloroeth
• 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.
• 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, for example, by the 3,4-dimethoxybenzyl or bis- (p-methoxyphenyl) methyl group, the hydrazide group, for example 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 the esterification are, for example, lower, optionally substituted alkanols such as methanol, ethanol, Cyanomethyl alcohol, benzoylmethyl alcohol or in particular tert-butanol, furthermore aralkanols such as aryl-lower alkanols, for example benzyl alcohols optionally substituted by lower alkyl or lower alkoxy groups or halogen atoms or benzhydroles such as benzhydrol, p-nitrobenzyl alcohol, p-methoxybenzyl alcohol, 2,4,6-trimethylbenzyl.
• alkanols such as methanol, ethanol, Cyanomethyl alcohol, benzoylmethyl alcohol or in particular tert-butanol
• aralkanols such as aryl-lower alkanols
• benzyl alcohols optionally substituted by lower alkyl or lower alkoxy groups or halogen atoms or benzhydroles
• alcohol phenols and thiophenols optionally substituted by electron-withdrawing substituents, such as thiophenol, thiocresol, p-nitrothiophenol, 2,4,5- 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.
• substituents such as thiophenol, thiocresol, p-nitrothiophenol, 2,4,5- 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 residues during the esterification are, in particular, residues derived from carbonic acid, such as benzoyloxycarbonyl or ethyloxycarbonyl.
• Groups suitable for etherification are e.g. Benzyl, tetrahydropyranyl or tert-butyl radicals.
• Also suitable for protecting the hydroxyl groups are those described in Ber. 100 (1967), 3838-3849 described 2,2,2-trifluoro-1-tert-butyloxycarbonylamino or -1-benzyloxycarbonylaminoethyl groups (Weygand).
• the tert-butyl ester group or the benzhydrol group is 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-l-tert-butyloxycarbonylamino-ethyl 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.
• 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 wherein R 1 9 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.
• protective groups can also be split off under basic conditions, their ability to split off under neutral conditions, in particular by the action of a salt of hydrofluoric acid, is of interest.
• 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) is carried out, for example, by reacting compound (VII) in the form of the activated carboxylic acid with (VIII), or by reacting the Acid (VII) with the compound (VIII), whose amino group 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-l, 2-dihydro-1-carbäthoxy- quinoline ester, N-hydroxypiperidine ester or an enol ester obtained with N-ethyl-5-phenyl-isoxazolium-3-sulfonate [W
• the amino group of the compound (VIII) can be activated, for example, by reaction with a phosphite.
• condensations are: the Weygand-WUnsch 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.
• these condensations can be carried out using the Merrifield method.
• 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 2 .
• 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 on the cysteine residue are different.
• 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 carried out in a manner known per se, for example according to one of those customary in peptide chemistry Methods in which amide or ester groups are converted.
• the 1-0-tosyl-2 (R) -0.3-0-isopropylidene glycerol thus obtained is condensed with N-acyl- (S) -cysteine in the presence of a basic agent, for example potassium carbonate, and the N- Acyl-S- [2 (R), 3-isopropylenedioxy-propyl] - (S) -cysteine, which is acidified, for example with acetic acid.
• a basic agent for example potassium carbonate
• N-acyl-S- [2 (R), 3-dihydroxypropyl] - (S) -cysteine obtained in this way 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.
• 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 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.
• the 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 crystals station. 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.
• an inert gas such as a nitrogen atmosphere.
• 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 starting material under the reaction conditions or in the form of a reactive one 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 corresponding to the formula (I) and those of the formula (VII), their mixtures, salts or complexes.
• Pharmaceutical preparations according to the invention 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. produced by means of conventional mixing, granulating, coating, dissolving or lyophilizing processes.
• Suitable carriers are in particular fillers, such as sugar, for example lactose, sucrose, mannitol or sorbitol, cellulose preparations and / or calcium phosphates, for example tricalcium phosphate or calcium hydrogenphosphate, and also binders, such as starch paste using, for example, corn, wheat, rice or potato starch, gelatin , Tragacanth, methyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose and / or polyvinyl pyrrolidone, and / or if desired.
• fillers such as sugar, for example lactose, sucrose, mannitol or sorbitol, cellulose preparations and / or calcium phosphates, for example tricalcium phosphate or calcium hydrogenphosphate
• binders such as starch paste using, for example, corn, wheat, rice or potato starch, gelatin , Tragacanth, methyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose
• Disintegrants such as the starches mentioned above, carboxymethyl starch, crosslinked polyvinylpyrrolidone, agar, alginic acid or a salt thereof, such as sodium alginate.
• Auxiliaries are primarily flow regulators and lubricants, for example silica, talc, stearic acid or salts thereof, such as magnesium or calcium stearate, and / or poly ethylene glycol.
• Dragee cores are provided with suitable, optionally gastric juice-resistant coatings, including 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 .
• suitable cellulose preparations such as acetyl cellulose phthalate or hydroxypropyl methyl cellulose phthalate, are used.
• Dyes or pigments can be added to the tablets or dragee coatings, for example for identification or for labeling different doses of active ingredient.
• the lipopeptide thus obtained shows in a thin layer chromatogram on silica gel derivative [N-palmitoyl-S-2 (R), 3-dihydroxypropyl] -Cys-Ser-Bu t ) -Ser (Bu t ) -Asn-OBu t an Rf value of 0.60 (chloroform-methanol 8: 2).
• 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, the remaining N-palmitoyl-cysteine is obtained by chloroform extraction, which is also recrystallized from gasoline.
• the ethyl acetate solution is evaporated and the Z-Ala-Glu (OBut) 2 with the Rf value 0.60 (in chloroform-methanol 95: 5) remains as foam, which is directly processed further: 8.5 g in 60 ml Dissolved methanol 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 (OBut) 2 is obtained as a white foam which has the Rf 0.36 in chloroform-methanol 9: 1.
• N-palmitoyl-S- [2 (R, S), 3-dihydroxypropyl] - (S) -cysteine With diphenyldiazomethane, as indicated above for the R compound, the N-palmitoyl-S- [2 ( R, S), 3-dipalmitoyloxypropyl] - (S) -cysteinbenzhydrylester: colorless crystals of mp. 84-85 °.
• 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
• the starting materials are, for example, N-myristoyl-S- [2 (R), 3-dihydroxypropyl] - (S) -cysteine, colorless crystals with a melting point of 140-150 °, and N-stearoyl-S- [2 (R) , 3-dihydroxypropyl] - (S) -cysteine, colorless crystals of mp 140-150 °; N-Lauroyl-S- [2 (R), 3-dihydroxypropyl] - (S) -cysteine, colorless crystals of mp 140-150 ?

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• 1978
  • 1978-06-14 DE DE7878100149T patent/DE2860893D1/de not_active Expired
  • 1978-06-14 EP EP78100149A patent/EP0000330B1/fr not_active Expired
  • 1978-06-16 FI FI781933A patent/FI781933A/fi not_active Application Discontinuation
  • 1978-06-16 CA CA000305645A patent/CA1139305A/fr not_active Expired
  • 1978-06-19 DK DK275178A patent/DK275178A/da not_active Application Discontinuation
  • 1978-06-19 AT AT0445178A patent/AT364095B/de not_active IP Right Cessation
  • 1978-06-19 DD DD78206107A patent/DD139257A5/xx unknown
  • 1978-06-19 IE IE1227/78A patent/IE46957B1/en unknown
  • 1978-06-19 PT PT68188A patent/PT68188A/pt unknown
  • 1978-06-19 ZA ZA00783504A patent/ZA783504B/xx unknown
  • 1978-06-19 AU AU37230/78A patent/AU525005B2/en not_active Expired
  • 1978-06-19 NZ NZ187611A patent/NZ187611A/xx unknown
  • 1978-06-19 IL IL54950A patent/IL54950A/xx unknown
  • 1978-06-19 NO NO782138A patent/NO148921C/no unknown
  • 1978-06-20 JP JP7382878A patent/JPS549224A/ja active Granted
  • 1978-06-20 PL PL21673978A patent/PL216739A1/xx unknown
  • 1978-06-20 GR GR56557A patent/GR71681B/el unknown
  • 1978-06-20 ES ES470961A patent/ES470961A1/es not_active Expired
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• 1979
  • 1979-03-16 ES ES478718A patent/ES478718A1/es not_active Expired
  • 1979-03-16 ES ES478717A patent/ES478717A1/es not_active Expired
• 1987
  • 1987-05-21 JP JP62122684A patent/JPS6372671A/ja active Granted

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