DE3811193A1 - Peptides, process for their preparation and pharmaceutical compositions containing these peptides - Google Patents

Abstract

Peptides of the general formula R1-U-X-Y-Z-Ile-Asn-Leu-Ile-Thr-X-W-X-Z-NH2 (the meaning of the various symbols is explained in the description) can be used for the therapy of cardiovascular disorders, coronary, cerebral and renal vasospasms as well as obesity.

Description

The invention relates to compounds of general formula I

R₁-U-X-Y-Z-Ile-Asn-Leu-Ile-Thr-X-W-X-Z-NH₂ (I)

their pharmaceutically acceptable salts, processes their preparation and containing these compounds pharmaceutical compositions.

In formula I stands

R₁ for R₂-CO or for a peptide-bound di-, tri-, tetra- or pentapeptide radical of the composition HZ-Pro-SXV, HZ-Pro-SV, HZ-Pro-V, HZV or acetyl Z-Pro-SXV-,
S for Ser, Glu, Cys or a bond,
U is an aliphatic amino acid such as Leu, Ile, Val, Ala or a bond,
X represents a basic amino acid such as Arg, Lys, Orn and Homo-Arg or a basic amino acid in which the amino group of the side chain is acetylated (eg Lys (Ac), Orn (Ac)), or a bond,
Y is an amino acid such as His, Trp, Phe and Tyr or a bond,
Z for an amino acid such as Tyr, Phe, His, Trp or an unnatural amino acid such. As naphthylalanine or substituted phenylalanine derivatives or if Z is not C-terminal for Cys,
W for an amino acid such as Gln, Asn, Glu, Asp,
V for a ω-aminoalkanoic acid radical such as ε-aminocaproic acid radical or a radical of the formula NH- (CH₂) _n -CO or NH- (CH₂) _n -CONH- (CH₂) _n CO with n = 2 to 15, or an amino acid sequence with turn structure such. Pro-Gly-T, Asn-Ser-T and Cys-Cys or Homo-Cys-Homo-Cys, which may each be disulphidically ring-closed with each other via their side chains,
T for one of the natural amino acids,
R₂ is C₁ to C₇ alkyl or benzyl.

If S for Glu and the non-C-terminal Z for Tyr stands, the side chain carboxyl group with the phenolic OH group of tyrosine ester-like be linked.

If S for Cys and the non-C-terminal Z for Cys can stand, the side chains of these amino acids be linked disulfide with each other.

In the sequence part U-X-Y-Z, amino acids are preferred, that in conjunction with the pentapeptide Ile-Asn-Leu-Ile-Thr alphahelical conformations favor.

Preference is given to compounds in which the symbols of the Formula I have the following meanings:

R₁: R₂-CO, HZ-Pro-SXV, HZ-Pro-SV, HZ-Pro-V, HZV or acetyl-Z-Pro-SXV-,
S: Ser or Glu,
U: Leu or a bond,
X: Arg, Lys, Orn or Homo-Arg,
Y: His or Trp,
Z: Tyr or Phe,
W: Gln or Asn,
V: ω- aminoalkanoic acid radical such as HN- (CH₂) _n -CO and NH- (CH₂) _n -CONH- (CH₂) _n -CO with n = 2 to 10,
R₂: for C₁ to C₇-alkyl or benzyl.

Possess compounds of general formula I. Acid groups, mainly carboxyl groups, or phenolic hydroxy groups and / or basic groups such as B. guanidino or amino functions. links of the general formula I can therefore either as internal salts, as salts with pharmaceutically acceptable Acids such as hydrochloric acid, sulfuric acid, phosphoric acid, Sulfonic acid or organic acids or as salts with pharmaceutically usable bases such as alkali or Alkaline earth metal hydroxides or carbonates, zinc or Ammonium hydroxides or organic amines such. B. Diethylamine, triethylamine, triethanolamine u. a. available.

Compounds of general formula I included Amino acids, either in their L-form or D-form can be present. Compounds of the formula I can therefore either from amino acid residues with L configuration can be constructed, or single or several of the amino acid residues of the compounds are in the D configuration.  

Preferred compounds of the invention are

1) H-Tyr-Pro-Ser-Lys-NH- (CH₂) ₅-CO-Arg-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
2) H-Tyr-Pro-Ser-Lys-NH- (CH₂) ₅-CO-Leu-Arg-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
3) H-Tyr-Pro-Ser-NH- (CH₂) ₇-CO-Arg-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
4) Ac-Tyr-Pro-Ser-Lys-NH- (CH₂) ₅-CO-Arg-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
5) H-Tyr-Pro-NH- (CH₂) ₁₀-CO-Arg-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
6) H-Tyr-Pro-NH- (CH₂) ₁₀-CO-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
7) H-Tyr-NH- (CH₂) ₁₀-CO-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Gln-Arg-Tyr-NH₂,
8) H-Tyr-Pro-Ser-NH- (CH₂) ₅-CO-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
9) H-Phe-Pro-Ser-Lys-NH- (CH₂) ₅-CO-Arg-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
10) H-Tyr-Pro-Ser-Lys-NH- (CH₂) ₇-CO-Arg-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
11) H-Tyr-Pro-Ser-NH- (CH₂) ₁₀-CO-Arg-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
12) H-Tyr-NH- (CH₂) ₁₂-CO-Arg-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
13) H-Tyr-Pro-Ser-Lys-NH- (CH₂) ₅-CO-Lys-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
14) H-Tyr-Pro-Ser-Lys-NH- (CH₂) ₅-CO-Lys-His-Tyr-Ile-Asn-Leu-Ile-Thr-Lys-Gln-Arg-Tyr-NH₂,
15) H-Tyr-Pro-Ser-Lys-NH- (CH₂) ₅-CO-Lys-His-Tyr-Ile-Asn-Leu-Ile-Thr-Lys-Gln-Lys-Tyr-NH₂,
16) H-Tyr-Pro-Ser-Lys-NH- (CH₂) ₅-CO-Arg-Trp-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
17) H-Tyr-Pro-Ser-Lys-NH- (CH₂) ₇-CO-Arg-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
18) H-Tyr-Pro-Ser-Lys-NH- (CH₂) ₅-CO-Arg-His-Phe-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
19) CH₃-CO-Arg-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
20) CH₃-CO-Lys-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂,
21) CH₃-CO-Lys-His-Tyr-Ile-Asn-Leu-Ile-Thr-Lys-Gln-Arg-Tyr-NH₂,
22) CH₃-CO-Lys-His-Tyr-Ile-Asn-Leu-Ile-Thr-Lys-Gln-Lys-Tyr-NH₂,
23) CH₃-CO-Lys (Ac) -His-Tyr-Ile-Asn-Leu-Ile-Thr-Lys (Ac) - Gln-Lys (Ac) -Tyr-NH₂,
24) CH₃-CO-Orn-His-Tyr-Ile-Asn-Leu-Ile-Thr-Orn-Gln-Orn-Tyr-NH₂.

The compounds of general formula I provide Segments, discontinuous interlinked Segments and analogues of the neuropeptide Y represent.

Neuropeptide Y (NPY) is one of 36 amino acids existing peptide, first in the brain of 1982 Pigs has been detected (K. Tatemoto et al., Nature 296, 659 [1982]). Meanwhile, NPY has been in all so far studied animal species and also in humans (M.M. Lundberg et al., Neurosci. Lett., 42, 167 [1983]). It is almost ubiquitous in the organism Before, particularly high concentrations are in the brain and in organs that are of the sympathetic nervous system innervated (heart, kidneys, adrenals, Vascular system), proven (T.L. O'Donohue et al., Peptides, 6, 755 [1985]). In many cases NPY is together with Norepinephrine (NA) located in the nerve endings and is released upon nerve impulse (J.M. Lundberg u. a., Regulatory Peptides, 13, 41 [1985]). Salient feature of exogenous in the Body periphery supplied NPY is a striking Increase in blood pressure. Together with NA, vasopressin (VP) and angiotensin II (AII) is one of the strongest NPY known autologous vascular contractors, d. H. hypertensive substances (P.C. Emson et al., TINS, 1984, 31). Local injection of NPY into the brain causes lower blood pressure (similar to adrenaline and NA) as well as a drastic increase in food intake (T.S. Gray et al., Life Sci., 38, 389 [1986]).  

The main attack of NPY in the periphery is that Heart (strong coronary constriction), kidneys (reduced blood flow) and the arterial vascular system (increase of peripheral resistance). In addition to his NPY also potentiates vasoconstrictor self-efficacy the effect of the above neurotransmitters (NA, VP, AII) as well as other body's own vasopressoric substances (L. Edvinsson et al., Br. Pharacol., 83, 519 [1984]).

From these effects as well as the accumulated occurrence of NPY in tissues relevant to blood pressure can be concluded that NPY at the Blood pressure regulation is involved.

Surprisingly, the inventive Compounds NPY-agonistic, but especially NPY antagonist activity.

An NPY agonist, d. H. hypertensive effect was given to anesthetized, despinalized rats demonstrated. After intravenous administration different doses of the respective test substance was the blood pressure increase registered and ED₂₀ values determined. The ED₂₀ value here denotes the dose the test substance which raises the blood pressure by 20 mm Hg increases.  

There were the following ED₂₀ values:

NPY 6.3 × 10 ^-10 M / kg Connection Example 1 5 × 10 ^-8 M / kg Connection Example 2 2 × 10 ^-8 M / kg Connection Example 19 2 × 10 ^-7 M / kg Connection Example 20 2 × 10 ^-7 M / kg

The NPY antagonist effect was anesthetized Rats determined. Was registered the inhibition of Blood pressure increase triggered by a defined dose of NPY after pretreatment with the test substance compared to the control values of non-pretreated Animals.

The following inhibition values were obtained at a dosage of 1 to 2 × 10 ^-6 M / kg test substance:

Compound Example 1 83%,
Compound Example 2 84%,
Compound Example 19 94%,
Compound Example 20 62%.

Because of the above-mentioned effectiveness can Compounds of the general formula I for therapy cardiovascular diseases such as hypertension, coronary, cerebral and renal vasospasm as well Obesity can be used. In addition, these can Compounds as valuable tools for generation and purification (affinity chromatography) of Antibodies and in RIA or ELISA assays Find.  

The invention therefore also relates to the use of Compounds of the invention as a remedy and pharmaceutical preparations containing these compounds contain. The application on humans is preferred. to parenteral administration, the compounds of the Formula I or its physiologically acceptable salts, possibly with the usual substances such as Solubilizers, emulsifiers or other auxiliaries brought in solution, suspension or emulsion. When Solvents come z. B. in question: water, physiological saline solutions or alcohols, eg. B. Ethanol, propanediol or glycerine, sugar solutions like Glucose or mannitol solutions or else a mixture from different solvents.

In addition, the compounds by implants, z. B. from polylactide, polyglycolide or Polyhydroxybutyric acid or intranasal preparations be applied.

The above-mentioned compounds may be general known methods of peptide chemistry, such as. In Houben-Weyl, Methods of Organic Chemistry, Vol. 15/2, described or preferred according to the Solid Phase Peptide Synthesis (e.g., B. Barany, R. B. Merrifield in The Peptides, Analysis, Synthesis, Biology, Vol. 2, 2-284 [1980], Academic Press, New York, or R.C. Sheppard, Int. J. Pept. Prot. Res., 21, 118 [1983]) or equivalent known methods getting produced. As amino protecting groups, the in Houben-Weyl, Methods of Organic Chemistry, Vol. 15/1, used, are preferred Urethane protective groups such. B. the Fluorenylmethoxycarbonyl or the tert-butyloxycarbonyl protecting group applied. To prevent Side reactions are usually present  functional groups in the side chains of the Amino acids by suitable protecting groups (see eg. T.W. Greene, Protective Groups in Organic Synthesis) additionally protected. Preferably be used Arg (NO₂), Arg (Mtr), Arg (di-Z), Arg (Pmc), Tyr (t-Bu), Tyr (Bzl), Tyr (Br-Z), Ser (t-Bu), Ser (Bzl), Glu (t-Bu), Glu (Bzl), Lys (BOC), Lys (Z), His (Trt), His (Bum), His (Dnp), His (Z), Thr (t-Bu), Thr (Bzl), Cys (Acm), Cys (4-Me-Bzl), Cys (t-Bu), Asp (t-Bu), Asp (Bzl), Orn (BOC), Orn (Z).

For the synthesis of the compounds of the general formula I according to the solid-phase synthesis are suitable as anchor groups all those which provide Peptidamide in the cleavage of the peptide from the polymeric carrier. In combination with the BOC-amino protecting group, preference is given to the benzhydrylamino anchor group (PG Pietta et al., J. Org. Chem., 39, 44 [1974]) and the 4-methylbenzhydrylamine anchor group (GR Matsueda et al., Peptides, 2, 45 [1981] ) used. In combination with the Fmoc protecting group different anchor groups are applicable:
p-aminomethyl-phenoxymethyl group (P. Pietta et al., J. Org. Chem., 40, 2995 [1975]),
Bis- and trialkoxybenzylamine linkers (F. Albericio, G. Barany, Int J. Pept. Prot. Res., 30, 206 [1987]) Benzhydryl alcohol linkers (G. Breipohl et al., Tetrah. Lett., 28, 5651 [1987]).

Especially the dimethoxyphenoxyvaleric acid anchor, the on alanine-loaded benzhydrylamine polystyrene over the carboxy group has been attached, has proven itself. To Coupling of the C-terminal Fmoc-protected amino acid with standard methods, e.g. DCCI or DIC / HOBt Room temperature overnight.  

After cleavage of the amino protecting group of the resin coupled amino acid with a suitable reagent, z. B. trifluoroacetic acid in dichloromethane in the case of BOC group or a 20-50% solution of piperidine in DMF in the case of the Fmoc group, which are suitable coupled with protected amino acids. This cycle will Run through one after the other as often as you want Resin-bound peptide has emerged.

For coupling, those known in peptide chemistry Methods (see Houben-Weyl, Methods of Organic Chemie, Vol. 15/2). Preferably used carbodiimides, such as. Dicyclohexylcarbodiimide, Diisopropylcarbodiimide or ethyl (3-dimethylaminopropyl) - carbodiimide or O-benzotriazole tetramethyl uronium hexafluorophosphate or benzotriazol-1-yl-oxy- tris- (dimethylamino) phosphonium hexafluorophosphate. By addition of 1-hydroxybenzotriazole (HOBt) or 3-hydroxy-4-oxo-3,4-dihydrobenzotriazine (HOObt) optionally suppresses the racemization or the Reaction rate can be increased. The Amino acids Asn and Gln are preferred in the form of their N-protected p-nitrophenyl ester coupled. The Clutches are usually equipped with a 2- to 5-fold excess of N-protected amino acid and Coupling reagent in solvents such as dichloromethane, Dimethylformamide, N-methylpyrrolidone (NMP) or Mixtures of these are performed. The course of the Coupling reaction is confirmed by the Kaiser test (E. Kaiser u. a., anal. Biochem., 34, 595 [1970]) or by the TNBS test tracked. If an incomplete Acylation is detected, the coupling is up to Completeness repeated. The solid phase synthesis can both manually and automatically with the help of a Peptidsynthesizers done.  

Preferred are solid phase syntheses using of the dimethoxyvaleric acid linker, Fmoc-amino protecting groups and the automatic synthesizer from ABI self-developed synthesis programs. It will Double clutches carried out; the first with DCC / HOBt in DMF, the second with symmetrical anhydrides in N-methylpyrrolidone. After the first clutch is a Washing step with E-ethylmorpholine in NMP interposed. After the second clutch takes place Capping with acetic anhydride and N-ethylmorpholine. The resin is washed with each coupling step DMF, NMP and isopropanol.

The part in the sequences of the present Invention occurring N-terminal acetyl group is by acetylating the peptide on the resin with acetic anhydride or acetylimidazole in DMF or NMP with eventual Addition of pyridine introduced as a base.

After construction of the peptides on the resin, the peptide with suitable reagents such as liquid hydrogen fluoride in Case of the built-up by the BOC strategy peptides or trifluoroacetic acid in the case of after Fmoc strategy constructed peptides from the polymeric carrier be split off. When using the above Anchor groups you get directly the corresponding Peptide amides. With these reagents are usually and using the appropriate side chain protecting groups also the functional, during the Synthesis protected groups of the peptide released.

Usually, in the cleavage of the peptide of the resin as cation scavenger substances such as phenol, Cresol, anisole, thiocresol, thioanisole, indole, Dimethyl sulfide or ethyl methyl sulfide or a Mixture of these substances to the hydrogen fluoride or added to the trifluoroacetic acid.  

The structure of the disulfide bridge occurs after cleavage the peptides from the resin by conventional methods, for. B. Houben-Weyl, Vol. 15/1, or B. Kamber u. a., Helv. Chim. Acta, 61, Fasc 4, 899 (1980), preferably with atmospheric oxygen or iodine in methanol or acetic acid.

The described homoarginine containing peptides are due to sales of the corresponding lysine derivatives made with O-methylisourea or by direct use suitably protected Homoarginine derivatives.

The purification of the crude products obtained takes place by gel chromatography z. To Sephadex G25 (MR <1400) or G15 (MR <1400) with 1% or 5% Acetic acid. If necessary, the others Purification by preparative RP-HPLC with methanol or acetonitrile-water gradient with the addition of 1 to 2% trifluoroacetic acid. For cleaning can also Sephadex or polystyrene based cation exchangers be applied.

analytics

The compounds of general formula I can be checked for purity by RP-HPLC. There is an amino acid analysis on each Ion exchanger (LKB) and on the gas chromatograph Chiral column for additional racemization control carried out. In addition, 13C NMR spectra (Bruker 400 MHz) and FAB mass spectra (MAT711, Varian). Testing for peptide amides takes place by means of resistance determination Carboxypeptidase Y.

The sequence determination is carried out by means of Gas-phase sequencing.  

The abbreviations used for the amino acids correspond to the usual three-letter code as z. B. in Europ. J. Biochem., 138, 9 (1984). The remaining abbreviations are explained below.

Ac = Acetyl Acm = Acetamidomethyl BOC = tert-butyloxycarbonyl t-Bu = tert-butyl bang = tert-butyloxymethyl Bzl = Benzyl Cl-Z = 4-chlorobenzyloxycarbonyl DCC = Dicyclohexylcarbodiimide DIC = Diisopropylcarbodiimide DMF = N, N-dimethylformamide dnp = 2,4-dinitrophenyl Fmoc = 9-fluorenylmethoxycarbonyl HOBt = 1-hydroxybenzotriazole 4-MeBzl = 4-methylbenzyl mtr = 4-methoxy-2,3,6-trimethylphenylsulfonyl mts = Mesitylene-2-sulfonyl pmc = 2,2,5,7,8-pentamethylchroman-6-sulfonyl pNP = 4-nitrophenyl Trt = Trityl

Example 1 H-Tyr-Pro-Ser-Lys-NH- (CH₂) ₅-CO-Arg- His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg- Tyr-NH₂

The synthesis was carried out with a peptide synthesizer Model 430A from Applied Biosystems under application the Fmoc strategy using the Dimethoxyvalerianoyloxybenzylamide anchor group (loading about 0.5 mmol / g resin), starting with the condensation of Fmoc-Tyr (t-Bu) -OH to the resin via DIC / HOBt in DMF (double clutch of 2 molar equivalents each). The Synthesis was via a modified control program carried out. Following amino acid derivatives were used: Fmoc-Arg (Mtr) -OH, Fmoc-Gln-OpNP, Fmoc-Thr (t-Bu) -OH, Fmoc-Ile-OH, Fmoc-Leu-OH, Fmoc-Asn-OH, Fmoc-His (Trt) -OH, Fmoc-NH- (CH₂) ₅-COOH, Fmoc-Lys (BOC) -OH, Fmoc-Ser (t-Bu) -OH, Fmoc-Pro-OH and BOC-Tyr (t-Bu) -OH.

The cleavage of the Fmoc protecting group was 25% Piperidine solution in DMF (2 × 10 min), subsequent washes with DMF. The first clutch each with Fmoc-amino acid / DIC / HOBt with 2 molar equivalents (30 min pre-activation), the Coupling time 60 or 90 min. The pre-activation time for Fmoc-Gln-OH / pNP-OH and Fmoc-Asn-OH was for 10 min reduced. Solvent was DMF, in the case of Fmoc-Gln-OpNP NMP. Thereafter, with DMF, NMP, Isopropanol and N-ethylmorpholine in NMP; the second coupling with the symmetrical anhydrides of Fmoc-amino acids were made in NMP. Then with DMF, Washed NMP and isopropanol and the cycle with the connected to the next amino acid. The splitting off of Peptides from the polymeric carrier were involved Trifluoroacetic acid (4h). The complete separation the Mtr-Schutzgruppe served the 30-minute treatment of the crude peptide with trifluoroacetic acid at 50 ° C. The Purification was by gel chromatography on Sephadex G15 carried out with 1% acetic acid. The clean Peptide was lyophilized.  

HPLC on Nucleosil C18.5 u, 4.6 x 250 mm; Detection at 220 nm gradient: 15% B to 60% B in 30 min, linear (A = water / trifluoroacetic acid 100: 0.1, B = acetonitrile / trifluoroacetic acid 100: 0.1) flow 1 ml / min;
Retention time 22.3 min;
Amino acid analysis: measured (theoretically)
Arg 3.00 (3), Asn 1.07 (1), Thr 1.04 (1), Gln 0.99 (1), Leu 0.93 (1), Ile 1.91 (2), His 1.03 (1), Tyr 2.77 (3), Pro 0.82 (1), Ser 0.86 (1), Lys 0.85 (1);
Racemization <3.5%;
no digestion with carboxypeptidase Y;
Gas phase sequencing: Tyr-Pro-Ser-Lys end of Edman degradation.

example 2 H-Tyr-Pro-Ser-Lys-NH- (CH₂) ₅-CO-Leu- Arg-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln- Arg-Tyr-NH₂

As described in Example 1, the synthesis was carried out.

HPLC: retention time 24.6 min;
Amino acid analysis:
Arg 3.00 (3), Asn 1.02 (1), Thr 1.05 (1), Gln 1.02 (1), Leu 2.11 (2), Ile 2.04 (2), His 0.94 (1), Tyr 2.71 (3), Pro 0.95 (1), Ser 1.05 (1), Lys 0.97 (1);
Racemization <3.4%;
no digestion with carboxypeptidase Y;
Gas phase sequencing: Tyr-Pro-Ser-Lys end of Edman degradation.

example 4 Ac-Tyr-Pro-Ser-Lys-NH- (CH₂) ₅-CO-Arg-His Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr- NH₂

As described in Example 1, the synthesis was carried out. Deviations: The coupling of the N-terminal tyrosine was reported as Fmoc-Tyr (t-Bu) -OH subsequent Fmoc cleavage and acetylation with Acetylimidazole in DMF (5 molar equivalents, 30 min) carried out.

HPLC: retention time 18.5 min;
Amino acid analysis:
Tyr 2.71 (3), Pro 0.96 (1), Ser 0.86 (1), Lys 1.00 (1), His 1.09 (1), Ile 1.90 (2), Leu 1.04 (1), Asn 0.97 (1), Thr 1.00 (1), Gln 0.95 (1), Arg 3.23 (3).

example 6 H-Tyr-Pro-NH- (CH₂) ₁₀-CO-Tyr-Ile-Asn- Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂

The synthesis was carried out as described in Example 1.

HPLC: retention time 23.4 min;
Amino acid analysis:
Tyr 2.53 (3), Pro 0.93 (1), Ile 1.96 (2), Leu 1.00 (1), Asn 0.99 (1), Thr 1.00 (1), Gln 1.02 (1), Arg 2.09 (2).

example 7 H-Tyr-NH- (CH₂) ₁₀-CO-Tyr-Ile-Asn-Leu-Ile- Thr-Arg-Gln-Arg-Tyr-NH₂

The synthesis was carried out as described in Example 1.

HPLC: retention time 24.9 min;
Amino acid analysis:
Tyr 3.26 (3), Ile 1.97 (2), Leu 1.05 (1), Asn 0.98 (1), Thr 1.00 (1), Gln 1.09 (1), Arg 2.17 (2).

example 8 H-Tyr-Pro-Ser-NH- (CH₂) ₅-CO-Tyr-Ile-Asn- Leu-Ile-Thr-Arg-Gln-Arg-Tyr-NH₂

The synthesis was carried out as described in Example 1.

HPLC: retention time;
Amino acid analysis:
Tyr 2.61 (3), Pro 1.10 (1), Ser 0.93 (1), Ile 1.90 (2), Leu 1.00 (1), Asn 1.04 (1), Thr 1.00 (1), Gln 1.00 (1), Arg 2.07 (2).

example 10 H-Tyr-Pro-Ser-Lys-NH- (CH₂) ₇-CO-Arg-His Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr- NH₂

The synthesis was carried out as described in Example 1.

HPLC: retention time 19.3 min;
Amino acid analysis:
Tyr 2.97 (3), Pro 0.90 (1), Ser 0.83 (1), Lys 0.94 (1), His 1.10 (1), Ile 1.92 (2), Leu 1.08 (1), Asn 0.97 (1), Thr 1.00 (1), Gln 1.03 (1), Arg 3.03 (3).

example 11 H-Tyr-Pro-Ser-NH- (CH₂) ₁₀-CO-Arg-His Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg- Tyr-NH₂

The synthesis was carried out as described in Example 1.

HPLC: retention time;
Amino acid analysis:
Tyr 2.70 (3), Pro 0.90 (1), Ser 0.90 (1), His 1.05 (1), Ile 2.00 (2), Leu 1.04 (1), Asn 0.99 (1), Thr 1.00 (1), Gln 1.04 (1), Arg 3.15 (3).

example 19 Ac-Arg-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg Gln-Arg-Tyr-NH₂

The synthesis was carried out analogously to that in Example 4 described way.

HPLC: retention time 25.6 min;
Amino acid analysis:
Thr 0.98 (1), Ile 1.82 (2), Leu 1.00 (1), Gln 1.09 (1), Tyr 1.86 (2), Arg 2.73 (3), Asn 1.00 (1), His 1.01 (1);
FAB-MS: M + H 1673.

example 20 Ac-Lys-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg Gln-Arg-Tyr-NH₂

The compound was analogous to that in Example 4 produced manner described.

HPLC: retention time 22.3 min;
Amino acid analysis:
Arg 2.00 (2), Tyr 2.06 (2), Gln 1.03 (1), Thr 1.07 (1), Ile 1.80 (2), Leu 0.99 (1), Asn 1.00 (1), His 0.98 (1), Lys 0.97 (1).

example 22 Ac-Lys-His-Tyr-Ile-Asn-Leu-Ile-Thr-Lys-Gln Lys-Tyr-NH₂

The compound was analogous to that in Example 4 received described manner.

HPLC: retention time 24.1 min;
Amino acid analysis:
Thr 0.95 (1), Ile 2.07 (2), Leu 1.01 (1), Gln 0.99 (1), Tyr 2.06 (2), Lys 3.03 (3), His 0.95 (1);
FAB-MS: M + H 1589.

example 23 Ac-Lys (Ac) -His-Tyr-Ile-Asn-Leu-Ile-Thr- Lys (Ac) -Gln-Lys (Ac) -Tyr-NH₂

The compound was synthesized analogously to the manner indicated in Example 4. After cleavage of the peptide from the resin, the ε- side chain amino groups were acetylated with acetylimidazoline in DMF.

HPLC: retention time 27.8 min;
FAB-MS: m + H 1589.

example 24 Ac-Orn-His-Tyr-Ile-Asn-Leu-Ile-Thr-Orn Gln-Orn-Tyr-NH₂

The compound was analogous to that in Example 4 received manner.

HPLC: retention time 24.1 min;
Amino acid analysis:
Thr 0.95 (1), Ile 1.93 (2), Leu 1.05 (1), Gln 0.99 (1), Tyr 2.00 (2), Orn 3.12 (3), Asn 1.00 (1), His 0.93 (1).

The other compounds mentioned above (compounds 3, 5, 9, 12-18 and 21) are prepared analogously.

injection Compound of Example 1 8 g methylparaben 5 g Polyparaben 1 g sodium chloride 25 g Water for injection 5 l

The compound, the preservatives and Sodium chloride is added to 3 liters of water for injection dissolved and to 5 l with water for injection  refilled. The solution is sterile filtered and Aseptically filled in pre-sterilized bottles with sterilized rubber caps are sealed. each Bottle contains 5 ml of solution.

Claims (9)

A compound of general formula I R₁-UXYZ-Ile-Asn-Leu-Ile-Thr-XWXZ-NH₂ (I) and its pharmaceutically acceptable salts, wherein
R₁ for R₂-CO or for a peptide-bound di-, tri-, tetra- or pentapeptide radical of the composition HZ-Pro-SXV, HZ-Pro-SV, HZ-Pro-V, HZV or acetyl Z-Pro-SXV-,
S for Ser, Glu, Cys or a bond,
U is an aliphatic amino acid such as Leu, Ile, Val, Ala or a bond,
X represents a basic amino acid such as Arg, Lys, Orn and Homo-Arg or a basic amino acid in which the amino group of the side chain is acetylated (eg Lys (Ac), Orn (Ac)), or a bond,
Y is an amino acid such as His, Trp, Phe and Tyr or a bond,
Z for an amino acid such as Tyr, Phe, His, Trp or an unnatural amino acid such. As naphthylalanine or substituted phenylalanine derivatives or if Z is not C-terminal for Cys,
W for an amino acid such as Gln, Asn, Glu, Asp,
V for a ω-aminoalkanoic acid radical such as ε-aminocaproic acid radical or a radical of the formula NH- (CH₂) _n -CO or NH- (CH₂) _n -CONH- (CH₂) _n CO with n = 2 to 15, or an amino acid sequence with turn structure such. Pro-Gly-T, Asn-Ser-T and Cys-Cys or Homo-Cys-Homo-Cys, which may each be disulphidically ring-closed with each other via their side chains,
T for one of the natural amino acids,
R₂ is C₁ to C₇ alkyl or benzyl
and wherein further
when S is Glu and the non-C-terminal Z is Tyr, the side-chain carboxyl group may be ester-linked to the phenolic OH group of tyrosine, and
if S stands for Cys and the non-C-terminal Z stands for Cys, the side chains of these amino acids can be linked disulfidically to one another. 2. A compound according to claim 1, wherein the symbols of the formula I have the following meanings:
R₁: R₂-CO, HZ-Pro-SXV, HZ-Pro-SV, HZ-Pro-V, HZV or acetyl-Z-Pro-SXV-,
S: Ser or Glu,
U: Leu or a bond,
X: Arg, Lys, Orn or Homo-Arg,
Y: His or Trp,
Z: Tyr or Phe,
W: Gln or Asn,
V: ω- aminoalkanoic acid radical such as HN- (CH₂) _n -CO and NH- (CH₂) _n -CONH- (CH₂) _n -CO with n = 2 to 10,
R₂: for C₁ to C₇-alkyl or benzyl. 3. A compound according to claim 1, wherein R₁ is the group CH₃CO means. 4. A compound according to claim 1, wherein in the definition R₁ Z is Tyr or Phe, S is Ser or a bond and X for Lys or a bond. 5. A compound according to claim 1 or 4, wherein V represents the group HN- (CH₂) _5-10 -CO-. 6. A compound according to any one of claims 1 to 5, wherein in the sequence -U-X-Y-Z-U for Leu or one Bond, X for Arg, Lys, Lys (Ac), Orn or one Bond, Y for His, Trp or a bond and Z for Tyr or Phe stands. 7. A compound according to any one of claims 1 to 6, wherein in the sequence -X-W-X-Z-X for Arg, Lys, Lys (Ac) or Orn, W stands for Gln and Z stands for Tyr.   8. A method for producing a compound according to one of claims 1 to 7 by stepwise Condensation of the respective amino acids or Amino acid sequences. 9. Pharmaceutical composition, characterized by the content of a compound according to any one of Claims 1 to 7.