DE3813989A1 - Peptides with phospholipase A2-inhibiting action - Google Patents

Abstract

Peptides with phospholipase A2-inhibiting action whose C-terminal alpha -carboxyl group is protected by an ester or amide functionality or can be reduced to the alcohol are described. The peptides of the formula L-B-A in which L can be absent or a lipophilic radical, B is a basic amino acid and A is an aromatic amino acid are used as medicines.

Description

The invention relates to peptides with phospholipase A₂-inhibiting activity, the C-terminal α- carboxyl group can be protected by an ester or amide function or reduced to alcohol.

Known peptide-like inhibitors of phospholipase A₂ are the lipocortins and plipastatins. Lipocortins that come from up to 346 amino acids exist (B.P. Wallner et al., Nature 320 [1986], 77-81; K.-S. Huang et al., Cell 46 [1986], 191-199; C. J. M. Saris et al., Cell 46 [1986], 201-212) antigenic properties (F.R. Hirata et al., Proc. Natl. Acad. Sci. USA 78 [1981], 3190) and therefore leave one therapeutic use appear questionable. The Plipastatine are cyclic decapeptides made by Bacillus cereus are formed (T. Nishikiori et al., J. of Antibiotics 39 [1986], 755-761). The activity of plipastatine was only for the phospholipase A₂ isolated from porcine pancreas described, their importance for inflammatory processes as not valid.

The invention is therefore based on the object, new Finding peptides that inhibit phospholipase A₂.

This object is achieved according to the invention by the peptides with phospholipase A₂-inhibiting activity, the C-terminal α- carboxyl group of which is protected by an ester function or an amide function, of the general formula I.

L-B-A (I)

in which

Or a lipophilic residue, Legs basic amino acid and An aromatic amino acid

mean, and their physiologically tolerable salts.  

Preferred radicals L, B and A are those which differ from naturally occurring amino acids (see e.g. Schröder, Lübke, The Peptides, Volume I, New York 1965, pages 137-270), derive their antipodes or their simple metabolites.

The radical L is in particular an N-terminal due to lipophilic acyl radicals, such as. B. (C₅-C₁₄) aryl- (C₁-C₄) alkyloxycarbonyl, (C₅-C₁₄) arylcarbonyl, (C₁-C₁₇) alkylcarbonyl; or protected by lipid residues consisting of up to 4 naturally occurring amino acids, lipophilic amino acid in the L or D configuration, which is acyl-like bound to the α- amino group of the basic amino acid or for a lipophilic acyl residue, such as. B. (C₅-C₁₄) aryl- (C₁-C₄) alkyloxycarbonyl, especially benzyloxycarbonyl, (C₅-C₁₄) aryl- (C₁-C₄) alkylcarbonyl, especially indole-3-butyryl, (C₅-C₁₄) - Arylcarbonyl, especially indole-2-carbonyl; or a (C₁-C₁₇) alkylcarbonyl radical, and their physiologically tolerable salts.

Such compounds of formula I are preferred, wherein

LIsoleucine, Leucine, Methionine, S-Protected Cysteine, Valine; phenylalanine, tryptophan or naphthylalanine optionally substituted by halogen, (C₁-C₄) alkyl or (C₁-C₄) alkoxy; on the hydroxy group substituted by (C₁-C₄) alkyl tyrosine, serine or threonine; histidine substituted on the N of the imidazole ring by (C₁-C₄) alkyl; in the side chain by (C₁-C₄) alkyl esterified or by lipophilic amines such as. B. (C₁-C₆) alkylamines or cyclohexylamine amidated aspartic acid, glutamic acid or aminoadipic acid; or on the ω- amino group with (C₁-C L) alkylcarbonyl substituted or unsubstituted lysine or ornithine; lipophilic residues, such as. B. benzyloxycarbonyl, indole-3-butyryl or indole-2-carbonyl, or (C₁-C₁₇) - alkylcarbonyl (fatty acid residue); Optionally alkylated on the guanidino group (C₁-C₆) - arginine or -homoarginine, lysine, ornithine, hydroxylysine or citrulline in the D or L configuration and optionally in the aromatic system by 1 to 5 identical or different radicals from the halogen series, (C₁ -C₄) alkyl or (C₁-C₄) alkoxy substituted phenylalanine, tryptophan, naphthylalanine, thienylalanine or tetrahydroisoquinoline-3-carboxylic acid; or tyrosine optionally substituted on the hydroxy group by (C₁-C₄) alkyl in the L or D configuration, the carboxyl groups of which are substituted by (C₁-C₄) alkyl or (C₅-C₁₄) aryl- (C₁-C₄) alkyl esters, are protected by the amide group, by primary (C₁-C₄) alkyl, (C₅-C₁₄) aryl (C₁-C₄) alkylamides and peptide residues consisting of up to 4 naturally occurring amino acids,

mean, and their physiologically tolerable salts.

In particular, salts with inorganic or organic acids such as B. HCl, HBr, H₂SO₄, H₃PO₄, Maleic acid, fumaric acid, citric acid, tartaric acid, Acetic acid in question. The chirality centers in the new Peptides can each have the R, S or R, S configuration have.

Alkyl can be straight-chain or branched. Corresponding applies to residues derived therefrom, such as. B. alkoxy and Alkylcarbonyl. For example, methyl and methoxy are preferred and methanoyl.

Under (C₅-C₁₄) aryl is also heteroaryl, such as. B. phenyl, Naphthyl, biphenylyl, fluorenyl, pyrrolyl, imidazolyl, Understand pyridyl and indolyl. Phenyl and are preferred Indole. The same applies to residues derived from it, such as B. aralkyl, aralkyloxycarbonyl and arylcarbonyl.  

Halogen is preferably fluorine, chlorine, bromine or iodine Chlorine or bromine.

(C₅-C₁₄) aryl (C₁-C₄) alkylamides are preferably derived from the biogenic amines, such as. B. phenethylamine, tryptamine, Serotonin, p-methoxyphenethylamine, 3,4-dimethoxyphenethylamine, p-hydroxyphenethylamine, 3,4-dihydroxyphenethylamine or Phenylalaninol.

L-B-A can also be part of a longer peptide or of a cyclopeptide, the structure L-B-A can occur in part or in whole several times.

The invention further relates to a method for Preparation of peptides of the formula I, which is characterized in that a fragment with a terminal Carboxyl group or its reactive derivative with a couples the corresponding fragment with a free amino group, if necessary to protect further functional groups (a) temporarily introduced protective group (s) split off and the compound thus obtained physiologically in it tolerated salt transferred.

The peptides according to the invention are synthesized according to general methods of peptide chemistry through gradual Construction from the C-terminal end or by segment condensation, as they e.g. B. in Wünsch et al., Houben-Weyl, Volume 15/1 and 2nd (Thieme, Stuttgart 1974) are described in detail. At the procedure using a segment coupling, care must be taken to use coupling methods with as little racemization as possible. In the examples here, Segment condensation especially the DCC / HOObt method used, according to previous experience with the lowest racemization during peptide synthesis.

The phospholipase A₂ uses arachidonic acid from phospholipids free (G.J. Blackwell and R.J. Flower, Br. Med. Bull. 39  [1983], 260-264). Since the arachidonic acid via the eicosanoid Cascade in connections such as B. prostaglandins and Leukotrienes can be transferred by inhibiting the Phospholipase A₂ the formation of these inflammatory effective arachidonic acid metabolites on an early Biosynthesis stage can be prevented. The invention Phospholipase A₂ inhibitors are therefore particularly there can be used therapeutically where prostaglandins, e.g. B. from E₂ type, leukotrienes, but also thromboxanes significantly on the Pathogenesis or maintenance of pathophysiological Processes are involved. Above all, this includes inflammatory, allergic, rheumatic and autoimmune diseases, such as B. bronchial asthma, Hay fever, drug allergy, urticaria, Skin diseases such as eczema and dermatitis, acute rheumatic fever, rheumatic inflammation of the joints, Muscle rheumatism. Other areas of application are Blood disorders such as acquired hemolytic anemia, idiopathic thrombopenia, agranulocytosis, myeloblastosis, Lymphogranulomatosis, hepatitis, ulcerative colitis, nephrotic syndrome, skin diseases such as pemphigus, Lupus erythematosus, dermatomyositis and treatment rejection after transplantation.

The invention therefore also relates to the use of Peptides of formula I as a medicament for the above Indications.

The peptides according to the invention were tested with the Phospholipase A₂ from polymorphonuclear leukocytes (PMN's). Since PMN's belong to the cell types that are inflammatory Processes are significantly involved (D. F. Bainton in G. Weissmann [ED.] "The Cell Biology of Inflammation", Volume 2, Page 1 ff., Elsevier / North Holland Biomedical Press, Amsterdam 1980), these findings are a special one Importance with regard to their therapeutic application  to. Isolated peritoneal rabbit PMN's used (Cunningham, J. Pathol. 128 [1979], 15-20), which in a concentration of 3 · 10⁸ cells / ml in distilled water and added as such for the Test were used. Determining the concentration, in which the phospholipase A₂ by the invention Connections to 50% is inhibited (IC₅₀ value), was after common methods using a series of concentrations carried out. For this purpose, the test substances were in 80 mmol / l Tris-HCl buffer (pH 7.5) dissolved. The calculation of the IC₅₀ Value is done with the help of a computer program according to the equation

in which [I] indicates the concentration of the inhibitor used.
Phospholipase A₂ inhibition

The substances are mainly applied parenterally, nasally or topically. But po application is also possible if suitable capsules prevent the sensitive substances in the stomach from being destroyed.

Examples 1. Z-Lys-Tyr-OMe 1a. Z-Lys (Boc) -Tyr (tBu) -OMe, C₃₃H₄₇N₃O₈ (MW 613.7)

13.95 g DCC (66 mmol) are added with stirring at 0 ° C a solution of 25.08 g (approx. 60-66 mmol) Z-Lys (Boc) -OH, 17.28 g (60 mmol) H-Tyr (tBu) -OMe · HCl and 7.78 ml N-ethylmorpholine in 120 ml of dimethylformamide. You leave Stir for 2 hours at 0 ° C and 6 hours at room temperature. It is left to stand at room temperature overnight, the is sucked Precipitation and the filtrate is concentrated in vacuo. The The residue is dissolved in ethyl acetate and successively with Water, saturated NaHCO₃ solution, KHSO₄ solution, NaHCO₃- Solution and water shaken. The ethyl acetate phase will dried over Na₂SO₄ and concentrated. The residue crystallizes from 200 ml of diisopropyl ether and 200 ml Petroleum ether.

Yield 30.95 g, mp 58-61 ° C, [ α ] = -6.8 ° ( c = 1, methanol).

1b. Z-Lys-Tyr-OMe acetate, C₂₆H₃₄N₃O₈ (MW 516.57)

2.5 g of Z-Lys (Boc) -Tyr (tBu) -OMe are 90% in 40 ml aqueous trifluoroacetic acid dissolved. Allow 2 hours Stand at room temperature and constricts in a vacuum. The residue is dissolved in water and with a weakly basic Ion exchanger in acetate form stirred until a pH from 4-5. The ion exchanger is filtered off and the filtrate is freeze-dried.

Yield 1.87 g.

For cleaning, 800 mg of the substance obtained are added a silica gel column in the eluent methylene chloride / Chromatographed methanol / acetic acid / water as 80: 20: 1: 1.  

The uniform fractions are combined and in vacuo constricted. The residue is dissolved in water and freeze-dried.

Yield 322 mg, [ α ] = + 4.2 ° ( c = 1, in 80% aqueous acetic acid). Peptide base content according to amino acid analysis: 88%.

2. Z-Trp-Lys-Tyr-OMe 2a. H-Lys (Boc) -Tyr (tBu) -OMe · HCl, C₂₅H₄₂Cl₁N₃O₆ (MW 516.08)

12.28 g (20 mmol) Z-Lys (Boc) -Tyr (tBu) -OMe are in Dissolved methanol and catalytically with hydrogen over Pd / C hydrated. The amino group released is approx. 1 N methanolic HCl using an autotitrator at pH 4.5 titrated. When the reaction is complete, the catalyst suction filtered and the filtrate concentrated. The arrears with Diethyl ether / petroleum ether (1: 1) treatment solid and can be sucked off.

Yield 9.31 g, mp. 150-152 ° C, [ α ] = + 16.9 ° ( c = 1, methanol).

2 B. Z-Trp-Lys (Boc) -Tyr (tBu) -OMe, C₄₄H₅₇N₅O₉ (MG 800)

To a solution of 6.0 g (10 mmol) of H-Lys (Boc) -Tyr (tBu) - OMe · HCl, 3.3 g (10 mmol) Z-Trp-OH and 1.86 g (10 mmol) HOBt in 50 ml of dimethylformamide are added at 0 ° C with stirring 2.31 g DCC. The procedure is now as in Example 1a. The Residue is made from diethyl ether / petroleum ether (1: 1) crystallized.

Yield 6.11 g.  

For cleaning, 200 g of silica gel in methylene chloride / Chromatographed methanol as 95: 5.

Yield 3.75 g, mp. 132 ° C, [ α ] = -12.9 ° ( c = 1, in methanol).

2c. Z-Trp-Lys-Tyr-OMe acetate, C₃₇H₄₅N₅O₉ (MG 703.8)

1.5 g Z-Trp-Lys (Boc) -Tyr (tBu) -OMe are mixed from 20 ml of 90% aqueous trifluoroacetic acid and 4 ml Dissolved 1,2-dimercaptoethane. Allow 1 hour Stand at room temperature and constricts in a vacuum. The residue is dissolved in water and with a weakly basic Ion exchanger in acetate form stirred until pH 3 has stopped. The ion exchanger is filtered off and the filtrate freeze-dried.

Yield 0.86 g.

For purification, the substance obtained above is in silica gel a mixture of 275 ml of water, 90.5 ml of acetic acid and 778 ml Chromatographed n-butanol. The unified factions are combined and concentrated in vacuo. The residue is dissolved in water and freeze-dried.

Yield 677 mg, [ a ] = -8.7 ° ( c = 1, in 10% acetic acid). Peptide base content according to amino acid analysis: 87%.

3. Z-Trp-Lys-Tyr-NH₂ 3a. Z-Lys (Boc) -Tyr (tBu) -OH, C₃₂H₄₅N₃O₈ (MW 599.7)

To a solution of 12.28 g (20 mmol) Z-Lys (Boc) -Tyr (tBu) - OMe in 75 ml of dioxane is added 25 ml of water and 21 ml of 1N NaOH. After 2.5 hours, the pH is brought to 4 with KHSO₄ solution placed and evaporated to dryness in vacuo. You distribute the residue between a KHSO₄ / K₂SO₄ solution and Ethyl acetate. The ethyl acetate phase is with water shaken out, dried over Na₂SO₄ and concentrated.  

The residue is triturated with diisopropyl ether firmly and can be vacuumed.

Yield 6.56 g, mp. 119-121 ° C with decomposition, [ α ] = + 3.3 ° ( c = 1, methanol).

3b. Z-Lys (Boc) -Tyr (tBu) -NH₂, C₃₂H₄₆N₄O₇ (MW 598.75)

To a solution of 5.99 g (10 mmol) of Z-Lys (Boc) -Tyr (tBu) -OH and 1.52 g (10 mmol) HOBt · NH₃ in 100 ml of dimethylformamide 2.47 g (12 mmol) of DCC are added with stirring at 0.degree. Man leaves 2 hours at 0 ° C and 2 hours at room temperature stir and stand for one day at room temperature. The The precipitate is filtered off and the filtrate in vacuo constricted. The residue is saturated with NaHCO₃ solution stirred. The precipitate is filtered off with KHSO₄ solution and washed water and dried over P₂O₅ in vacuo.

Yield 5.21 g.

For cleaning, the substance is warm in 30 ml of methanol dissolved and precipitated with 20 ml of water (ice bath). The Precipitation is filtered off and dried.

Yield 4.87 g.

For further purification, the substance is extracted from ethyl acetate / Methyl tert-butyl ether reprecipitated.

Yield 4.3 g, mp 153 ° C, [ α ] = -22.4 ° ( c = 1, in methanol).

3c. H-Lys (Boc) -Tyr (tBu) -NH₂ · HCl, C₂₄H₄₁Cl₁N₄O₅ (MG 501.07)

3 g of Z-Lys (Boc) -Tyr (tBu) -NH₂ are analogous to Example 2a catalytically hydrogenated. The residue is mixed with diethyl ether rubbed and vacuumed.

Yield 2.14 g, mp. 187-189 ° C with decomposition, [ α ] = + 18.5 ° ( c = 1, methanol).

3d. Z-Trp-Lys (Boc) -Tyr (tBu) -NH₂, C₄₃H₅₆N₆O₈ (MW 784.9)

To a solution of 1.35 g (4 mmol) Z-Trp-OH, 2.0 g (4 mmol) H-Lys (Boc) -Tyr (tBu) -NH₂ · HCl, 0.57 g (4 mmol) HOBt and 0.52 ml (4 mmol) N-ethylmorpholine in 25 ml dimethylformamide 0.93 g (4.5 mmol) of DCC are added with stirring at 0.degree. Man works as in example 1a. The backlog is in 20 ml of methanol dissolved and precipitated with 20 ml of water. The Precipitation is suctioned off and in a vacuum over P₂O₅ dried.

Yield 2.05 g, mp. 158-160 ° C with decomposition, [ α ] = -14.3 ° ( c = 1, methanol).

3e. Z-Trp-Lys-Tyr-NH₂ acetate, C₃₆H₄₄N₆O₈ (MW 688.78)

1.5 g of Z-Trp-Lys (Boc) -Tyr (tBu) -NH₂ are analogous to Example 2c implemented and cleaned.

Yield 851 mg, [ α ] = + 1.5 ° ( c = 1, 10% aqueous acetic acid). Peptide base content according to amino acid analysis: 87%.

4. Z-Lys-Tyr-Phe-OMe 4a. Z-Tyr (tBu) -Phe-OMe, C₃₁H₃₆N₂O₆ (MG 532.6)

To a solution of 40.7 g Z-Tyr (tBu) -OH, 21.6 g H-Phe- OMe · HCl, 13.5 g HOBt and 13 ml N-ethylmorpholine in 200 ml Dimethylformamide is added at 0 ° C with stirring 22 g DCC. One works up as in experiment 1a. The arrears with Rubbed petroleum ether, suction filtered and dried in vacuo.

Yield 50.3 g (94.4%), mp. 103-104 ° C, [ α ] = -12.3 ° ( c = 1, in methanol).

4b. H-Tyr (tBu) -Phe-OMe · HCl, C₂₃H₃₁Cl₁N₂O₄ (MW 434.9)

49.5 g of Z-Tyr (tBu) -Phe-OMe are analogous in methanol Example 2a catalytically hydrogenated.

Yield 49 g of a greasy substance that is not crystallized.

4c. Z-Lys (Boc) -Tyr (tBu) -Phe-OMe, C₄₂H₅₆N₄O₉ (MW 760.9)

To a solution of 21.5 g H-Tyr (tBu) -Phe-OMe · HCl, 6.67 g HOBt and 6.43 ml N-ethylmorpholine in 150 ml Dimethylformamide is added at 0 ° C with stirring 27.67 g Z-Lys (Boc) -OTcp. The mixture is left at 0 ° C. for 1 hour and 2 hours Stir at room temperature and concentrate. Is worked up analogous to example 1a. The residue is in a little ethyl acetate dissolved and precipitated with petroleum ether. The precipitation is suctioned off and dried in vacuo.

Yield 29.1 g (77.4%), mp. 93-94 ° C, [ α ] = -19.2 ° ( c = 1, in methanol).

4d. Z-Lys-Tyr-Phe-OMe, C₃₃H₄₀N₄O₇ (MG 604.7)

0.8 g of Z-Lys (Boc) -Tyr (tBu) -Phe-OMe are analogous to Example 2c implemented.

Yield 0.44 g, [ α ] = -18.2 ° ( c = 1, in methanol). Peptide base content according to amino acid analysis: 81%.

5. H-Lys-Tyr-Phe-OMe 5a. H-Lys (Boc) -Tyr (tBu) -Phe-OMe · HCl, C₄₃H₅₁Cl₁N₄O₇ (MW 663.2)

28 g of Z-Lys (Boc) -Tyr (tBu) -Phe-OMe are analogous to Example 2a catalytically hydrogenated. The residue is mixed with diethyl ether triturated, suction filtered and dried in vacuo.  

Yield 22.4 g (92%), mp. 145-147 ° C, [ α ] = + 18.2 ° ( c = 1, in methanol).

5b. H-Lys-Tyr-Phe-OMe acetate, C₂₇H₃₈N₄O₇ (MW 530.6)

0.4 g of H-Lys (Boc) -Tyr (tBu) -Phe-OMe · HCl become analogous Example 2c implemented.

Yield 293 mg, [ α ] = + 16.0 ° ( c = 1, in methanol). Peptide base content according to amino acid analysis: 66%.

6. Z-Trp-Lys-Tyr-Phe-OMe 6a. Z-Trp-Lys (Boc) -Tyr (tBu) -Phe-OMe, C₅₃H₆₆N₆O₁₀ (MG 947.16)

To a solution of 1.58 g (4.7 mmol) Z-Trp-OH, 3.11 g (4.7 mmol) H-Lys (Boc) -Tyr (tBu) -Phe-OMe · HCl, 0.64 g (4.7 mmol) HOBt and 0.6 ml (4.7 mmol) N-ethylmorpholine in 35 ml Dimethylformamide is added at 0 ° C with stirring 1.07 g (5.2 mmol) DCC. Now proceed as in Example 1a. The residue is dissolved in methanol and with water like. The precipitate is filtered off and in vacuo dried over P₂O₅.

Yield 5.21 g, mp 158-159 ° C, [ α ] = -25.5 ° ( c = 1, in dimethylformamide).

6b. Z-Trp-Lys-Tyr-Phe-OMe acetate, C₄₆H₅₄N₆O₁₀ (MW 850.9)

0.947 g (1 mmol) of Z-Trp-Lys (Boc) -Tyr (tBu) -Phe-OMe implemented analogously to Example 2c. The arrears with Methyl tert-butyl ether triturated and suction filtered. The Compound is recrystallized from methanol.

Yield 0.28 g.

All of the substance obtained above is in 1.5 ml Acetic acid dissolved, diluted with 150 ml of water and freeze-dried.  

[ α ] = -1.7 ° ( c = 1, in water), content of peptide base according to amino acid analysis: 88%.

7. Z-Lys-Tyr-Phe-NH₂ 7a. Z-Tyr (tBu) -Phe-NH₂, C₁₀H₃₅N₃O₅ (MW 517.6)

To a solution of 19.9 g (53 mmol) Z-Tyr (tBu) -OH, 10 g (50 mmol) H-Phe-NH₂ · HCl, 6.8 g (50 mmol) HOBt and 6.4 ml (50 mmol) of N-ethylmorpholine in 75 ml of dimethylformamide 11.33 g (55 mmol) of DCC at 0 ° C. with stirring. You leave Stir at 0 ° C for 1 hour and then at room temperature stand overnight. The precipitate is suctioned off and that Concentrated filtrate. The residue becomes saturated NaHCO₃ solution stirred. The precipitate is suctioned off and washed with water. The still moist substance will boiled with 150 ml of methanol and refrigerated overnight.

Yield 23.1 g, mp. 182 ° C, [ α ] = -3.6 ° ( c = 1, in acetic acid).

7b. H-Tyr (tBu) -Phe-NH₂ · HCl, C₂₂H₃₀Cl₁N₃O₃ (MW 419.96)

22.43 g of Z-Tyr (tBu) -Phe-NH₂ are analogous to Example 2a catalytically hydrogenated. The residue is mixed with diethyl ether rubbed.

Yield 17.45 g, mp. 130-140 ° C with decomposition, [ α ] = + 36.1 ° ( c = 1, in acetic acid).

7c. Z-Lys (Boc) -Tyr (tBu) -Phe-NH₂, C₄₁H₅₅N₅O₈ (MW 745.9)

To a solution of 16 g (42 mmol) Z-Lys (Boc) -OH, 16.76 g (40 mmol) H-Tyr (tBu) -Phe-NH₂ · HCl, 6.56 g (40 mmol) HOObt and 5.12 ml (40 mmol) of N-ethylmorpholine in 100 ml  Dimethylformamide is added at 0 ° C with stirring 9.27 g (45 mmol) DCC. The approach is analogous to Example 7a worked up. It is recrystallized from dry ethanol.

Yield 21.41 g, mp. 208 ° C, [ α ] = -34.7 ° ( c = 1, in dimethylformamide).

7d. Z-Lys-Tyr-Phe-NH₂, C₃₂H₃₉N₅O₆ (MG 589.7)

3.73 g (5 mmol) of Z-Lys (Boc) -Tyr (tBu) -Phe-NH₂ are analogous Example 2c implemented. The residue is dissolved in water and adjusted to pH 6 with NaHCO₃ solution. The aqueous solution is shaken out with ethyl acetate and then with NaCl saturated. Now crystallization occurs.

Yield 2.41 g.

It is recrystallized from absolute ethanol.

Yield 1.82 g, [ α ] = -9.7 ° ( c = 1, in 80% aqueous acetic acid). Peptide base content according to amino acid analysis: 52%.

8. Z-Trp-Lys-Tyr-Phe-NH₂ 8a. H-Lys (Boc) -Tyr (tBu) -Phe-NH₂ · HCl, C₃₃H₄₉Cl₁N₅O₆ (MW 647.25)

14 g of Z-Lys (Boc) -Tyr (tBu) -Phe-NH₂ are analogous to Example 2a catalytically hydrogenated.

Yield 10.7 g, mp. 203-205 ° C with decomposition, [ α ] = + 12.8 ° ( c = 1, in methanol).

8b. Z-Trp-Lys (Boc) -Tyr (tBu) -Phe-NH₂, C₅₂H₆₅N₇O₉ (MW 932.15)

To a solution of 2.7 g (8 mmol) Z-Trp-OH, 5.1 g (8 mmol) H-Lys (Boc) -Tyr (tBu) -Phe-NH₂ · HCl, 1.31 g (8 mmol) HOObt and 1.02 ml (8 mmol) N-ethylmorpholine in 50 ml dimethylformamide  are added at 0 ° C and with stirring 1.85 g (9 mmol) DCC. Man allows to stir at 0 ° C for 1 hour and provides overnight Room temperature. The precipitate is suctioned off and that The filtrate was concentrated in vacuo. The residue is in 100 ml Dissolved ethyl acetate. Add 100 ml of diethyl ether and turns cold. The precipitate is filtered off and in vacuo dried.

Yield 6.11 g, mp. 186 ° C. with decomposition, [ a ] = -32.6 ° ( c = 1, in dimethylformamide).

8c. Z-Trp-Lys-Tyr-Phe-NH₂, C₄₃H₄₉N₇O₇ (MW 775.9)

2.0 g of Z-Trp-Lys (Boc) -Tyr (tBu) -Phe-NH₂ are analogous Example 2c implemented. The residue is mixed with diethyl ether rubbed and vacuumed. The substance becomes saturated NaHCO₃ solution triturated and stirred for 1 hour. The The precipitate is filtered off and recrystallized from ethanol.

Yield 0.98 g, [ α ] = -7.3 ° ( c = 1, in 80% aqueous acetic acid). Peptide base content according to amino acid analysis: 70%.

9. H-Trp-Lys-Tyr-Phe-NH₂-diacetate, C₄₂H₅₁N₇O₉ (MW 797.9)

500 mg of Z-Trp-Lys-Tyr-Phe-NH₂ in 90% aqueous Acetic acid catalytically hydrogenated over Pd / C. After finished Hydrogenation, the catalyst is suctioned off and the filtrate constricted. The residue is triturated with diethyl ether and vacuumed. Then the substance with ethanol stirred, suction filtered and with an ethanol / diethyl ether Mixture washed and dried in vacuo.

Yield 300 mg, [ α ] = + 6.5 ° ( c = 1, in 80% acetic acid). Peptide base content according to amino acid analysis: 74%.

10. Z-D-Trp-Lys-Tyr-Phe-NH₂ 10a. Z-D-Trp-Lys (Boc) -Tyr (tBu) -Phe-NH₂, C₅₂H₆₅N₇O₉ (MW 932.15

Is produced as in Example 8b. Mp 192-194 ° C, [ α ] = -21.3 ° ( c = 1, in dimethylformamide).

10b. Z-D-Trp-Lys-Tyr-Phe-NH₂, C₄₃H₄₉N₇O₇ (MW 775.9)

2 g of Z-Trp-Lys (Boc) -Tyr (tBu) -Phe-NH₂ are analogous to Example 8c implemented.

Yield 0.98 g, [ α ] = -7.3 ° ( c = 1, in 80% aqueous acetic acid), content of peptide base according to amino acid analysis: 70%.

11a. Z-D-Trp-Lys (Boc) -OH, C₃₀H₃₈N₄O₇ (MW 566.6)

To a suspension of 19.71 g H-Lys (Boc) -OH and 11.38 g HOBt in 160 ml of dimethylformamide is added at room temperature with stirring 41.43 g Z-D-Trp-OTcp. After 4 hours of stirring everything is solved at room temperature. One leaves overnight stand and constrict in a vacuum. The backlog becomes one 3-stage countercurrent distribution in 400 ml portions between ethyl acetate and semi-saturated NaHCO₃ solution subject. The 3 ethyl acetate fractions are combined and shaken with KHSO₄ / K₂SO₄ solution and water, over Na₂SO₄ dried and concentrated. The arrears with Rubbed petroleum ether, suction filtered and dried.

Yield 55.6 g.

For further purification, the substance is added to 600 g of silica gel chromatographed. First, only with methylene chloride  eluted, later with a mixture of methylene chloride / Methanol / water like 80: 20: 5.

Yield 45.2 g, mp. 94 ° C with decomposition, [ α ] = + 0.7 ° ( c = 1, in methanol).

11b. H-D-Trp-Lys (Boc) -OH acetate, C₂₄H₃₆N₄O₇ (MW 492.6)

41 g of Z-D-Trp-Lys (Boc) -OH become more aqueous in 1500 ml of 90% Acetic acid dissolved and catalyzed over Pd / C with hydrogen hydrated. When the reaction is complete, the catalyst suction filtered and the filtrate concentrated. The arrears with Triturated ether, suction filtered and dried in vacuo.

Yield 32 g (89%), [ α ] = -52.4 ° ( c = 1, methanol).

11c. Z-Phe-D-Trp-Lys (Boc) -OHcyclohexylamine, C₄₅H₆₀N₆O₈ (MW 813.0)

To a solution of 31.53 g of H-D-Trp-Lys (Boc) -OH acetate and 9.11 g HOBt in 150 ml dimethylformamide are added Room temperature with stirring 25.37 g Z-Phe-ONSu. You stir 4 hours at room temperature and leaves overnight Stand at room temperature. The next day you put 400 ml of water and 64 ml of saturated NaHCO₃ solution. The precipitation is suctioned off and washed well with water and over P₂O₅ dried.

Yield 45.4 g (99.4%).

The substance obtained above is between ethyl acetate, which one add a little n-butanol and distribute the KHSO₄ / K₂SO₄ solution. The ethyl acetate / n-butanol phase is dried over Na₂SO₄ and constricted. The residue is dissolved in ethyl acetate and mixed with 7.3 ml of cyclohexylamine. One poses overnight at 4 ° C and sucks off the precipitation the next day. The Precipitation is again with 350 ml of ethyl acetate  boiled, cooled, suction filtered and dried.

Yield 39.5 g (75.9%), mp. 148-152 ° C with decomposition, [ α ] = + 9.6 ° ( c = 1, in methanol).

11d. H-Phe-D-Trp-Lys (Boc) -OH, C₃₁H₄₁N₅O₆ (MW 579.7)

39 g of Z-Phe-D-Trp-Lys (Boc) -OH · cyclohexylamine are between Ethyl acetate and 48 ml of 1 N H₂SO₄ distributed under ice cooling. The ethyl acetate phase is with KHSO₄ / K₂SO₄ solution and water shaken out, dried over Na₂SO₄ and concentrated. The The residue is dissolved in 600 ml of 90% acetic acid and Catalytically hydrogenated over Pd / C with hydrogen. To completed reaction, the catalyst is suctioned off and that Concentrated filtrate. The residue is triturated with ether vacuumed and dried.

Yield 28.7 g.

The substance is recrystallized from isopropanol.

Yield 21.85 g (78.6%), mp. 142-144 ° C with decomposition, [ α ] = + 41 ° ( c = 1, in methanol).

11e. Fmoc-Cys (StBu) -Phe-D-Trp-Lys (Boc) -OH, C₅₃H₆₄N₆O₉S₂ (MW 993.3)

To a suspension of 5.8 g of H-Phe-D-Trp-Lys (Boc) -OH and 1.42 g of HOBt in 20 ml of dimethylformamide are added Room temperature with stirring 6.11 g Fmoc-Cys (StBu) -OTcp. Man stir for 4 hours at room temperature and leave overnight Stand at room temperature. The mixture is concentrated in vacuo and the residue dissolved in ethyl acetate and successively with Water, saturated NaHCO₃ solution, KHSO₄ / K₂SO₄ solution and Shaken out water, dried with Na₂SO₄ and concentrated. The residue is triturated with petroleum ether and suction filtered.

Yield 8.18 g.  

For cleaning, the substance obtained above is over 100 g Chromatographed silica gel. As an eluent first methylene chloride and then a mixture of Methylene chloride / methanol as 80:20 used. The uniform fractions are pooled, in vacuo concentrated and the residue triturated with petroleum ether and aspirated.

Yield 6.42 g (64.6%), mp. 126-132 ° C with decomposition, [ α ] = -68.4 ° ( c = 1, in methanol).

11f. Fmoc-Cys (StBu) -Thr-ol, C₂₆H₃₄N₂O₅S₂ (MW 518.7)

To a solution of 5.36 g (20 mmol) H-Thr-ol · HOObt in 35 ml of dimethylformamide are added at 0 ° C with stirring 12.2 g Fmoc-Cys (StBu) -OTcp. The mixture is left at 0 ° C. for 1 hour and 4 hours stir at room temperature, concentrate in vacuo and dissolve the Residue in ethyl acetate. The ethyl acetate phase will successively with water, saturated NaHCO₃ solution, KHSO₄ / K₂SO₄ solution and water shaken out over Na₂SO₄ dried and concentrated. The substance crystallizes from isopropanol. The substance is sucked off and with Washed petroleum ether.

Yield 6.63 g, mp. 155 ° C with decomposition, [ α ] = -86.5 ° ( c = 1, in methanol).

11g. H-Cys (StBu) -Thr-ol · HOObt, C₁₈H₂₉N₅O₅S₂ (MW 459.6)

To a solution of 6.47 g (12.5 mmol) of Fmoc-Cys (StBu) -Thr-ol 12.87 ml (125 mmol) are added to 55 ml of dimethylformamide Diethylamine. The mixture is left to stand at room temperature for 5 minutes and constricts in a vacuum. The residue is in methanol solved. The insoluble is filtered off. The filtrate is 2.03 g (12.5 mmol) of HOObt were added and the mixture was concentrated. The  The residue is triturated with diethyl ether, suction filtered and dried in vacuo.

Yield 5.25 g, amorphous, [ α ] = -26.3 ° ( c = 1, in methanol).

11h. Fmoc-Tyr (Et) -Cys (StBu) -Thr-ol, C₃₇H₄₈N₃O₇S₂ (MW 710.9)

To a solution of 4.31 g (10 mmol) Fmoc-Tyr (Et) -OH and 4.58 g (10 mmol) H-Cys (StBu) -Thr-ol · HOObt in 40 ml Dimethylformamide is added at 0 ° C with stirring 2.26 g (11 mmol) DCC. It now continues as in Example 1a method. The residue is triturated with diethyl ether, suction filtered and dried in vacuo.

Yield 5.61 g.

Chromatograph the substance for purification Silica gel. The mixture is eluted first Methylene chloride / methanol like 95: 5 and later with Methylene chloride / methanol as 90:10. The uniform Fractions are pooled and concentrated in vacuo. The The residue is triturated with diethyl ether, suction filtered and dried.

Yield 4.5 g, mp. 126 ° C, [ α ] = -73.4 ° ( c = 1, in methanol).

11i. H-Tyr (Et) -Cys (StBu) -Thr-ol, C₂₂H₃₆N₃O₆S₂ (MG 503.2)

To a solution of 4.35 g (6 mmol) Fmoc-Tyr (Et) -Cys (StBu) - Thr-ol in 20 ml of dimethylformamide is added at room temperature 6.2 ml (60 mmol) of diethylamine with stirring. Allow 5 minutes Stir at room temperature, constricts and rubs the Residue with diethyl ether.

Yield 2.68 g (88.7%), mp. 113 ° C, [ α ] = -69.8 ° ( c = 1, in methanol).

11k. Fmoc-Cys (StBu) -Phe-D-Trp-Lys (Boc) -Tyr (Et) -Cys (StBu) -Thr-ol, C₇₅H₉₉N₉O₁₃S₄ (MG 1462.7)

To a solution of 4.26 g (4.3 mmol) Fmoc-Cys (StBu) - Phe-D-Trp-Lys (Boc) -OH, 2.16 g (4.3 mmol) H-Tyr (Et) -Cys (StBu) - Thr-ol and 0.72 g (4.3 mmol) HOB in 15 ml dimethylformamide are added at 0 ° C with stirring 0.93 g (4.5 mmol) DCC and continues to work as in Example 7a. The raw substance is triturated with diethyl ether, suction filtered, dried and then reprecipitated from 60 ml of methanol and 30 ml of water. The precipitate is filtered off and vacuum over P₂O₅ dried.

Yield 5.26 g (83.7%).

11l. H-Cys (StBu) -Phe-D-Trp-Lys (Boc) -Tyr (Et) -Cys (StBu) -Thr-ol, C₆₀H₈₉N₉O₁₁S₄ (MG 1240.5)

To a solution of 5.26 g (3.59 mmol) Fmoc-Cys (StBu) - Phe-D-Trp-Lys (Boc) -Tyr (Et) -Cys (StBu) -Thr-ol in 15 ml Dimethylformamide is added at room temperature with stirring 14.36 ml (35.9 mmol) diethylamine-dimethylformamide mixture. After 5 minutes, the mixture is concentrated in vacuo and the residue triturated with diethyl ether. The precipitate is suctioned off and dried.

Yield 3.93 g, mp. 145-147 ° C with decomposition, [ α ] = -81.8 ° ( c = 1, in dimethylformamide).

11m. Boc-D-Phe-Cys (StBu) -Phe-D-Trp-Lys (Boc) -Tyr (Et) - Cys (StBu) -Thr-ol, C₇₄H₁₀₆N₁₀O₁₄S₄ (MG 1487.7)

To a solution of 0.87 g (3.3 mmol) of Boc-D-Phe-OH, 3.7 g (3 mmol) H-Cys (StBu) -Phe-D-Trp-Lys (Boc) -Tyr (Et) -Cys (StBu) - Thr-ol and 0.54 g (3.3 mmol) HOObt in 15 ml Dimethylformamide is added at 0 ° C with stirring 0.72 g (3.5 mmol) DCC and works analogously to Example 7a. The  Raw substance is made from 20 ml of methanol with 20 ml of water like.

Yield 4.29 g, mp. 212-213 ° C with decomposition, [ α ] = -66.2 ° ( c = 1, in dimethylformamide).

To a solution of 4.16 g (2.9 mmol) of Boc-D-Phe-Cys (StBu) - Phe-D-Trp-Lys (Boc) -Tyr (Et) -Cys (StBu) -Thr-ol in 50 ml Trifluoroethanol is added to 3.5 ml of tri-n-butylphosphine. The mixture is stirred for 4 hours at room temperature and concentrated in Vacuum on. The residue is mixed with ethyl acetate / petroleum ether rubbed and vacuumed.

Yield 2.24 g.

The substance obtained above (1.71 mmol) is in 150 ml Acetic acid dissolved and with stirring at room temperature a mixture of 1500 ml acetic acid, 34.52 ml 0.1 N Iodine solution, 500 ml water and 471 mg sodium acetate trihydrate dripped. Stir for 5 minutes and decolorises excess iodine with aqueous ascorbic acid Solution. The mixture is concentrated in vacuo and the Rub the residue with water. The precipitation will suction filtered, washed well with water and over P₂O₅ in Vacuum dried.

Yield 2.07 g.

For further cleaning, 1 g of the above is obtained Substance on silica gel in methylene chloride / methanol like 9: 1 chromatographed. The unified factions are combined and concentrated in vacuo.

Yield 680 mg.  

680 mg obtained above are analogous to Example 2c treated. The crude diacetate (yield 451.3 mg) is for purification by distribution chromatography ®Sephadex LH 20 chromatographed. Here will ®Sephadex LH 20 with a mixture of 275 ml water, 90.5 ml of acetic acid and 778 ml of n-butanol saturated. As The eluent is a mixture of 4300 ml water, 430 ml of n-butanol and 350 ml of acetic acid.

Yield 306 mg, [ α ] = -65.8 ° ( c = 1, in 1% acetic acid).

12. Z-Phe-Lys-Phe-Tyr-OMe 12a. Z-Phe-Tyr-OMe, C₂₇H₂₈N₂O₆ (MG 476.5)

To a solution of 14.95 g (50 mmol) of Z-Phe-OH, 11.58 g (50 mmol) H-Tyr-OMe · HCl, 7 g (50 mmol) HOBt and 6.4 ml N-ethylmorpholine in 150 ml of dimethylformamide is added 0 ° C and with stirring 11.33 g (55 mmol) DCC. The approach is worked up analogously to Example 1a. The residue is rubbed with petroleum ether.

Yield 21.2 g (89%), mp. 127 ° C, [ α ] = -15.1 ° ( c = 1, in methanol).

12b. H-Phe-Tyr-OMe · HCl, C₁₉H₂₃Cl₁N₂O₄ (MW 378.8)

9.52 g (20 mmol) of Z-Phe-Tyr-OMe are analogous to Example 2a catalytically hydrogenated. The residue is mixed with diethyl ether rubbed.

Yield 7.05 g, mp. 128-131 ° C, [ α ] = + 7.6 ° ( c = 1, in methanol).

12c. Z-Lys (Boc) -Phe-Tyr-OMe, C₃₈H₄₈N₄O₉ (MG 704.8)

To a solution of 6.84 g (18 mmol) of Z-Lys (Boc) -OH, 6.82 g (18 mmol) H-Phe-Tyr-OMe · HCl, 2.43 g (18 mmol) HOBt and  2.3 ml (18 mmol) N-ethylmorpholine in 60 ml dimethylformamide are added at 0 ° C with stirring 4.2 g DCC. The approach is going worked up analogously to Example 1a. The arrears with Diethyl ether triturated, suction filtered and dried.

Yield 4.48 g, mp. 98 ° C, [ α ] = -9.2 ° ( c = 1, in methanol).

12d. H-Lys (Boc) -Phe-Tyr-OMe · HCl, C₃₀H₄₃Cl₁N₄O₇ (MG 607.16)

4.3 g of Z-Lys (Boc) -Phe-Tyr-OMe are analogous to Example 2a catalytically hydrogenated. The residue is mixed with diethyl ether rubbed.

Yield 3.65 g, mp. 132 ° C, [ α ] = -4.0 ° ( c = 1, in methanol).

12e. Z-Phe-Lys (Boc) -Phe-Tyr-OMe, C₄₇H₅₇N₅O₁₀ (MW 852.0)

To a solution of 1.79 g (6 mmol) Z-Phe-OH, 3.64 g (6 mmol) H-Lys (Boc) -Phe-Tyr-OMe · HCl, 0.81 g (6 mmol) HOBt and 0.77 ml of N-ethylmorpholine in 35 ml of dimethylformamide 1.35 g (6.6 mmol) of DCC are added at 0 ° C. with stirring. The Approach is worked up analogously to Example 7a. The Crude substance is dissolved in 30 ml of methanol and with 30 ml Water like.

Yield 6.19 g, mp. 151 ° C, [ α ] = -10.6 ° ( c = 1, in dimethylformamide).

12f. Z-Phe-Lys-Phe-Tyr-OMe acetate, C₄₄H₅₃N₅O₁₀ (MW 811.9)

0.852 g (1 mmol) of Z-Phe-Lys (Boc) -Phe-Tyr-OMe become analogous Example 2c treated. The residue is treated with methyl tert. triturated butyl ether and then from methanol / water recrystallized.

Yield 0.34 g.  

The substance obtained above is dissolved in 2 ml of acetic acid. The solution is mixed with 150 ml of water and freeze-dried.

Yield 231 mg, mp. 205 ° C, [ α ] = -19.8 ° ( c = 1, in dimethylformamide).

Amino acid analysis: Phe 2.0, Tyr 0.78, Lys 1.0; Content of Peptide base according to amino acid analysis: 91%.

13. H-Lys-Phe-Leu-Lys-Phe-OtBu 13a. Fmoc-Lys (Z) -Phe-OtBu, C₄₂H₄₇N₃O₇ (MG 705.86)

To a solution of 5.03 g (10 mmol) of Fmoc-Lys (Z) -OH, 2.6 g (10 mmol) H-Phe-OtBu · HCl, 1.35 g (10 mmol) HOBt and 1.3 ml (10 mmol) of N-ethylmorpholine in 50 ml of dimethylformamide 2.2 g of DCC at 0 ° C. with stirring. The approach becomes analog Example 1a worked up. The arrears with Diethyl ether triturated, suction filtered and dried.

Yield 6.2 g [ α ] = -6.74 ° ( c = 1, in 90% aqueous acetic acid).

13b. Fmoc-Lys (Z) -Phe-OH, C₃₈H₃₉N₃O₇ (MW 649.75)

2.95 g of Fmoc-Lys (Z) -Phe-OtBu become 90% in 30 ml aqueous trifluoroacetic acid dissolved. Allow 1 hour Stand at room temperature and constricts in a vacuum. The residue is triturated with diethyl ether and suction filtered.

Yield 2.32 g.

13c. Fmoc-Leu-Lys (Z) -Phe-OtBu, C₄₈H₅₈N₄O₈ (MG 819.02)

To a solution of 3.17 g (4.5 mmol) of Fmoc-Lys (Z) -Phe-OtBu 4.7 ml (45 mmol) are added to 20 ml of dimethylformamide Diethylamine. The mixture is stirred for 10 minutes at room temperature and constricts in a vacuum. The backlog will be in little Dissolved methylene chloride, filtered and on  Silica gel cleaned. One uses as eluent first methylene chloride to remove the impurities to dissolve and later to elute the tripeptide Mixture of methylene chloride / methanol as 95: 5.

Yield 2.3 g of oily substance.

To a solution of 1.58 g (4.5 mmol) Fmoc-Leu-OH, above recovered 2.3 g H-Leu-Lys (Z) -Phe-OtBu and 607.5 mg HOBt in 20 ml of dimethylformamide are added at 0 ° C with stirring 940 mg DCC. The batch is worked up analogously to Example 1a. The residue is triturated with diethyl ether and suction filtered and dried in vacuo.

Yield 2.7 g.

13d. Fmoc-Lys (Z) -Phe-Leu-Lys (Z) -Phe-OtBu, C₇₁H₅₈N₇O₁₃ (MW 1244.5)

To a solution of 2.67 g (3.26 mmol) of Fmoc-Leu-Lys (Z) - Phe-OtBu in 20 ml dimethylformamide is added Room temperature 3.42 ml (32.6 mmol) of diethylamine and stirred 10 minutes at room temperature. You work analogously Example 13c.

Yield 2 g of oily substance.

To a solution of 2.2 g (3.27 mmol) of Fmoc-Lys (Z) -Phe-OH, 2 g of oil obtained above and 533 mg HOObt in 20 ml Dimethylformamide is given 720 mg DCC at 0 ° C. The approach is worked up analogously to Example 7a.

Yield 3.14 g, mp. 178-182 ° C, [ α ] = -20.8 ° ( c = 1, in 90% aqueous acetic acid).

13e. H-Lys-Phe-Leu-Lys-Phe-OtBu triacetate, C₄₆H₇₅N₇O₁₂ (MW 918.1)

1 g of Fmoc-Lys (Z) -Phe-Leu-Lys (Z) -Phe-OtBu in 20 ml 90% aqueous acetic acid dissolved and over Pd / C catalytically hydrogenated. When the reaction is complete, the  Aspirated catalyst and the filtrate concentrated. The The residue is triturated with diethyl ether and suction filtered.

Yield 750 mg.

The substance is purified chromatographically on silica gel. A mixture of 450 ml serves as eluent Methylene chloride, 200 ml of methanol, 150 ml of methyl glycol, 100 ml water and 5 g ammonium acetate.

Yield 450 mg, [ α ] = -25.5 ° ( c = 1, in water).

Amino acid analysis: Leu 1.0; Phe 1.96; Lys 2.03; salary on peptide base according to amino acid analysis: 62%.

14. Ac-Trp-Lys-Trp-NH- (CH₂) ₄-CH₃ 14a. Z-Trp-n-pentylamide, C₂₄H₂₉N₃O₃ (MW 407.5)

To a solution of 16.9 g (50 mmol) Z-Trp-OH, 4.35 g (50 mmol) n-pentylamine and 7 g HOBt in 100 ml Dimethylformamide is added at 0 ° C with stirring 11.33 g (55 mmol) DCC. The procedure is analogous to example 1a. The Residue crystallizes from diethyl ether / petroleum ether like 1: 1.

Yield 18.35 g (90%), mp. 146 ° C, [ α ] = -22.3 ° ( c = 1, in dimethylformamide).

14b. H-Trp-n-pentylamide · HCl, C₁₆H₂₄Cl₁N₃O₁ (MW 309.85)

16.28 g (40 mmol) Z-Trp-n-pentylamide are analogous Example 2a catalytically hydrogenated. The residue crystallizes from diethyl ether / petroleum ether.

Yield 12.28 g (99%), mp 152 ° C, [ a ] = + 44.1 ° ( c = 1, in methanol).

14c. Z-Lys (Boc) -Trp-n-pentylamide, C₃₅H₄₉N₅O₆ (MW 635.8)

To a solution of 6.2 g (20 mmol) of H-Trp-n-pentylamide · HCl, 7.6 g (20 mmol) Z-Lys (Boc) -OH, 2.7 g HOBt and 2.56 ml  (20 mmol) N-ethylmorpholine in 70 ml of dimethylformamide 4.73 g (23 mmol) of DCC are added at 0 ° C. with stirring. The Approach is worked up analogously to Example 1a. The Substance crystallizes from diethyl ether / petroleum ether.

Yield 11 g (86.6%), mp. 126 ° C, [ α ] = -14.1 ° ( c = 1, in dimethylformamide).

14d. H-Lys (Boc) -Trp-n-pentylamide · HCl, C₂₇H₄₄Cl₁N₅O₄ (MW 538.1)

9.52 g (15 mmol) of Z-Lys (Boc) -Trp-n-pentylamide analogously to Example 2a catalytically hydrogenated. The residue crystallizes from diethyl ether.

Yield 5.91 g, mp 100-128 ° C with decomposition, [ α ] = + 23.6 ° ( c = 1, in methanol).

14e. Z-Trp-Lys (Boc) -Trp-n-pentylamide, C₄₆H₅₉N₇O₇ (MG 822)

To a solution of 5.38 g (10 mmol) of H-Lys (Boc) -Trp-n- pentylamide · HCl, 3.38 g (10 mmol) Z-Trp-OH, 1.4 g (10 mmol) HOBt and 1.28 ml (10 mmol) N-ethylmorpholine in 40 ml Dimethylformamide is added at 0 ° C while stirring 2.47 g (12 mmol) DCC. The approach is analogous to Example 1a worked up. The raw substance is made from ethanol / water fell over, suction filtered and dried over P₂O₅.

Yield 5.63 g, mp. 188-190 ° C with decomposition, [ α ] = -22.7 ° ( c = 1, in dimethylformamide).

14f. H-Trp-Lys (Boc) -Trp-n-pentylamide · HCl, C₃₈H₅₄Cl₁N₇O₅ (MW 724.36)

3.7 g (4.5 mmol) of Z-Trp-Lys (Boc) -Trp-n-pentylamide analogously to Example 2a catalytically hydrogenated. The residue crystallizes from diethyl ether.  

Yield 3.04 g (93%), mp. 200-202 ° C with decomposition, [ α ] = + 4.2 ° ( c = 1, in methanol).

14g. Ac-Trp-Lys (Boc) -Trp-n-pentylamide, C₄₀H₄₄N₆O₆ (MW 704.84)

To a solution of 2.89 g (4 mmol) of H-Trp-Lys (Boc) -Trp-n- pentylamide · HCl and 0.51 ml (4 mmol) N-ethylmorpholine in 25 ml of dimethylformamide are added at 0 ° C with stirring 0.7 g Acetic acid N-hydroxysuccinimide ester. One leaves overnight stand at room temperature and constrict. The backlog will triturated with water, suction filtered and dried over P₂O₅. The substance is dissolved in 75 ml of hot methanol and mixed with 40 ml of water in the heat. It is cooled to 0 ° C from. Here the substance crystallizes out.

Yield 2.52 g, mp. 192-194 ° C with decomposition, [ α ] = -20.3 ° ( c = 1, in dimethylformamide).

14h. Ac-Trp-Lys-Trp-n-pentylamide triacetate, C₄₁H₅₉N₇O₁₀ (MW 809.9)

1 g Ac-Trp-Lys (Boc) -Trp-n-pentylamide are analogous to Example 2c implemented. The residue is treated with methyl tert-butyl ether rubbed and vacuumed. Then the substance with NaHCO₃- The solution is triturated, suction filtered and washed with water. The The substance thus obtained is in 50 ml of 3% acetic acid solved. The insoluble is filtered off and the filtrate freeze-dried.

Yield 798.3 mg, [ α ] = + 21.5 ° ( c = 1, in water), content of peptide base according to amino acid analysis: 71%.

15a. Fmoc-Trp-Cys (StBu) -Thr-ol, C₃₇H₄₄N₄₅O₆S₂ (MG 704.9)

To a solution of 2.13 g Fmoc-Trp-OH and 2.3 g H-Cys (StBu) - Thr-ol · HOObt (see Example 11g) in 15 ml of dimethylformamide  are given at 0 ° C, 1.13 g DCC. The approach becomes analog Example 1a worked up. The arrears with Rubbed petroleum ether, suction filtered and dried in vacuo.

Yield 3.13 g.

For cleaning the substance is 30 minutes with diethyl ether stirred, suction filtered and dried.

Yield 2.23 g (63%), mp. 101-111 ° C, [ a ] = -77.5 ° ( c = 1, methanol).

15b. Fmoc-Cys (StBu) -Phe-D-Trp-Lys (Boc) -Trp-Cys (StBu) -Thr-ol, C₇₅H₉₆N₁₂O₁₂S₄ (MG 1485.95)

To a solution of 1.97 g (2.8 mmol) Fmoc-Trp-Cys (StBu) - Thr-ol in 28 ml of dimethylformamide is added Room temperature with stirring 12.2 ml (28 mmol) diethylamine Dimethylformamide solution. Stir for 5 minutes Room temperature and concentrated in vacuo. The backlog will Chromatographed over 60 g of silica gel analogously to Example 13c.

Yield 0.9 g (66.6%) of oily substance.

To a solution of the 0.9 g H-Trp-Cys (StBu) obtained above - Thr-ol, 1.85 g Fmoc-Cys (StBu) -Phe-D-Trp-Lys (Boc) -OH (see Example 11e) and 0.31 g HOB in 6 ml dimethylformamide 0.41 g of DCC are added at 0 ° C. with stirring. The approach is going worked up analogously to Example 7a.

Yield 2.58 g (93.4%).

For cleaning, the substance obtained above is on silica gel chromatographed. Is eluted with a mixture of Methylene chloride / methanol as 92: 8. The uniform Fractions are concentrated in vacuo and the residue triturated with petroleum ether.

Yield 1.8 g (65.2%), mp. 180-182 ° C with decomposition, [ α ] = -113.9 ° ( c = 1, in methanol).

15c. H-Cys (StBu) -Phe-D-Trp-Lys (Boc) -Trp-Cys (StBu) -Thr-ol, C₆₀H₈₆N₁₀O₁₀S₄₄ (MG 1235.7)

To a solution of 1.64 g Fmoc-Cys (StBu) -Phe-D-Trp- Lys (Boc) -Trp-Cys (StBu) -Thr-ol (1.1 mmol) in 11 ml Dimethylformamide is added 4.4 ml (11 mmol) of diethylamine Dimethylformamide solution. Stir for 5 minutes Room temperature and concentrated in vacuo. The backlog will Chromatographed on silica gel as in Example 13c. The uniform fractions are pooled, in vacuo concentrated and triturated with petroleum ether.

Yield 0.74 g (54.4%), mp 144-148 ° C, [ α ] = -119.9 ° ( c = 1, in methanol).

To a solution of 0.41 g Boc-D-Phe-OH, 0.62 g H-Cys (StBu) - Phe-D-Trp-Lys (Boc) -Trp-Cys (StBu) -Thr-ol and 0.071 g HOBt in 3 ml of dimethylformamide are added at 0 ° C with stirring 0.11 g DCC. The batch is worked up analogously to Example 7a.

Yield 0.74 g.

For cleaning, the substance obtained above is on silica gel chromatographed. Is eluted with a mixture of Methylene chloride / methanol as 93: 7.

Yield 350 mg.

To a solution of 0.34 g (0.23 mmol) of the above Boc-D-Phe-Cys (StBu) -Phe-D-Trp-Lys (Boc) -Trp-Cys (StBu) -Thr-ol 0.58 ml is added to 5 ml of trifluoroethanol at room temperature (2.3 mmol) tri-n-butylphosphine. Stir for 4 hours Room temperature and constricts. The arrears with a Mixture of ethyl acetate / petroleum ether triturated and suction filtered.

Yield 290 mg.  

281 mg of the Boc-D-Phe-Cys-Phe-D-Trp-Lys (Boc) obtained above - Trp-Cys-Thr-ol are dissolved in glacial acetic acid. This solution leaves quickly with stirring at room temperature to a mixture from 215 ml of glacial acetic acid, 71.6 ml of water, 58.8 mg of sodium acetate and 4.4 ml of 0.1 N iodine solution are added dropwise. You still leave Stir for 5 minutes and decolorize the solution with a aqueous ascorbic acid solution. The solution is concentrated and the residue triturated with water, suction filtered and over P₂O₅ dried. The substance is on silica gel chromatographed. Is eluted with a mixture of Methylene chloride / methanol as 95: 5. The concentrated Fractions are triturated with petroleum ether.

Yield 120 mg.

110 mg of the above

are implemented analogously to Example 2c. The backlog is between Ethyl acetate and water distributed. The ethyl acetate phase will shaken again with water. The United Water phases are with a weakly basic Ion exchanger in acetate form stirred until a pH of 3-4. The ion exchanger is filtered off and the filtrate freeze-dried.

Yield 65 mg.

Amino acid analysis: Cys 1.83; Phe 2.0; Lys 1.0; Trp 1.0 (was determined via the UV spectrum); Peptide base content according to amino acid analysis: 55%.

16. H-Arg-Tyr (tBu) -OtBu 16a. Z-Arg (Z₂) -Tyr (tBu) -OtBu, C₄₇H₅₇N₅O₁₀ (MG 852.0)

To a solution of 2.89 g (5 mmol) Z-Arg (Z₂) -OH, 1.65 g H-Tyr (tBu) -OtBu · HCl, 0.68 g HOBt and 0.65 ml N-ethylmorpholine in 20 ml DMF is added with stirring at 0 ° C 1.1 g DCC.  

The batch is worked up analogously to Example 1a. The Residue is in for purification on silica gel Chromatograph methylene chloride / methanol as 95: 5. The uniform fractions are combined and in vacuo constricted. The residue is triturated with petroleum ether.

Yield 2.57 g, mp 123-124 ° C, [ α ] = + 9.6 ° ( c = 1, in glacial acetic acid).

16. H-Arg-Tyr (tBu) -OtBu.2 HCl, C₂₃H₄₁Cl₂N₅O₄ (MW 522.5)

2.04 g of Z-Arg (Z₂) -Tyr (tBu) -OtBu are analogous to Example 2a catalytically hydrogenated. The residue is mixed with diethyl ether rubbed.

Yield 1.06 g, amorphous, [ a ] = + 22.6 ° ( c = 1, methanol).

17. Fmoc-Cys (StBu) -Arg-Phe-OtBu 17a. Z-Arg (Z₂) -Phe-OtBu, C₄₃H₄₉N₅O₉ (MG 779.9)

To a solution of 17.3 g (30 mmol) of Z-Arg (Z₂) -OH, 7.73 g H-Phe-OtBu · HCl, 4.9 g HOObt and 3.9 ml N-ethylmorpholine in 70 ml of dimethylformamide are added with stirring at 0 ° C 6.6 g DCC. The procedure is analogous to example 1a. The The residue is triturated with diethyl ether, suction filtered and dried in vacuo.

Yield 19 g, [ α ] = -0.5 ° ( c = 1, in 80% aqueous acetic acid).

17b. H-Arg-Phe-OtBu.2 HClO₄, C₁₉H₃₃Cl₂N₅O₁₁ (MW 578.4)

18.6 g of Z-Arg (Z₂) -Phe-OtBu are dissolved in 200 ml of dimethylacetamide dissolved and hydrogenated catalytically over Pd / C with hydrogen. With the help of an autotitrator during the hydrogenation maintained by adding 1N HClO₄ pH 5. After finished Hydrogenation, the catalyst is suctioned off and the filtrate  constricted. The residue is dissolved in water, over a Filtration layer filtered and freeze-dried.

Yield 21.1 g of oil.

The oil is triturated twice with diethyl ether Decanted diethyl ether and the oily residue in vacuo dried.

Yield 18.5 g (135%, so still contains solvent).

17c. Fmoc-Cys (StBu) -Arg-Phe-OtBu acetate, C₄₃H₅₈N₆O₈S₂ (MW 851.1)

To a solution of 4.31 g (10 mmol) Fmoc-Cys (StBu) -OH, 7.74 g H-Arg-Phe-OtBu.2 HClO₄ (oily obtained above Substance), 1.35 g HOBt and 1.3 ml N-ethylmorpholine in 40 ml of dimethylformamide are added at 0 ° C with stirring 2.2 g DCC. The batch is worked up analogously to Example 1a. The The residue is triturated with diethyl ether and dried.

Yield 8 g.

1.3 g of the substance obtained above are in 30 ml 55% acetic acid dissolved and over a weakly basic Ion exchanger chromatographed in acetate form. Man eluted with 55% acetic acid. The eluate is concentrated taken up in water and freeze-dried.

Yield 1.05 g, [ α ] = -45.5 ° ( c = 1, in methanol), content of peptide base according to amino acid analysis: 79%.

18. Fmoc-Arg-Pro-Cys (StBu) -Arg-Phe-OtBu 18a. Fmoc-Arg-Pro-OtBu, C₃₀H₃₉N₅O₅ (MG 549.68)

To a solution of 5.95 g (15 mmol) Fmoc-Arg-OH, 2.57 g H-Pro-OtBu, 2.02 g HOBt and 2.7 g pyridinium perchlorate in  30 ml of dimethylformamide are added at 0 ° C with stirring 3.3 g DCC. The procedure is analogous to example 1a. The residue is triturated with diethyl ether and suction filtered.

Yield 9.4 g.

750 mg of those obtained above are used for characterization Substance dissolved in 40% aqueous acetic acid and over a weakly basic ion exchanger in acetate form chromatographed. The eluate is concentrated and freeze-dried.

Yield 690 mg Fmoc-Arg-Pro-OtBu acetate, C₃₂H₄₃N₅O₇ (MW 609.7), [ α ] = -44.2 ° ( c = 1, in methanol, substance content according to amino acid analysis: 93%.

18b. Fmoc-Arg-Pro-OH, C₂₆H₃₁N₅O₅ (MW 493.57)

8.5 g of raw substance obtained above is 90% in 85 ml aqueous trifluoroacetic acid dissolved. Allow 1 hour Stand at room temperature and constricts in a vacuum. The residue is triturated with diethyl ether and suction filtered.

Yield 7.9 g.

7.16 g of the substance obtained above are in 100 ml of water suspended and with a saturated NaHCO₃ solution a pH Value of 6 set. It is set at 4 ° C over the weekend and sucks off the precipitate and dries over P₂O₅ in Vacuum.

Yield 6.15 g, [ α ] = -32.2 ° ( c = 1, in 80% aqueous acetic acid.

18c. H-Cys (StBu) -Arg-Phe-OtBu, C₂₆H₄₄N₆O₄S₂ (MG 568.8)

To a solution of 17.84 g (20 mmol) of Fmoc-Cys (StBu) -Arg- Phe-OtBu · HClO₄ (manufactured analogously to raw substance from Example 17c) in 100 ml of dimethylformamide is added  Room temperature 21 ml (200 mmol) diethylamine. You stir 10 minutes at room temperature and concentrated in vacuo. The The residue is triturated twice with diethyl ether and decanted. The oily residue is dried in a high vacuum.

Yield 14.6 g.

18d. Fmoc-Arg-Pro-Cys (StBu) -Arg-Phe-OtBu diacetate, C₅₆H₈₁N₁₁O₁₂S₂ (MG 1164.46)

To a solution of 4.94 g (10 mmol) of Fmoc-Arg-Pro-OH, 5.7 g (10 mmol) H-Cys (StBu) -Arg-Phe-OtBu, 163 mg HOObt and 1.8 g pyridinium perchlorate in 100 ml dimethylformamide 2.2 g of DCC are added at 0 ° C. The procedure is analogous to example 1a on. The residue is triturated with diethyl ether.

Yield 8.7 g.

500 mg of the substance obtained above are on silica gel chromatographed. The following serves as eluent Mixture: 450 ml methylene chloride, 200 ml methanol, 150 g Acetic acid up to a pH of 6.8.

Yield 341 mg, [ α ] = -60.8 ° ( c = 1, water), content of peptide base according to amino acid analysis: 77%.

19. H-Arg-Pro-Cys (StBu) -Arg-Phe-OtBu triacetate, C₄₃H₇₅N₁₁O₁₂S₂ (MG 1002.27)

To a solution of 3.3 g Fmoc-Arg-Pro-Cys (StBu) -Arg-Phe- OtBu in 20 ml of dimethylformamide gives 3.32 ml of diethylamine. The mixture is left to stand at room temperature for 10 minutes and concentrated in Vacuum on. The residue is triturated with diethyl ether and vacuumed.

Yield 2.7 g.

The substance obtained above is analogous to Example 18d cleaned chromatographically.  

Yield 1.23 g, [ α ] = -42.4 ° ( c = 1, in water), content of peptide base according to amino acid analysis: 71%.

20. Z-Trp-Asp (OtBu) -Arg-Phe-NH₂ 20a. Z-Arg (Z₂) -Phe-NH₂, C₃₉H₄₂N₆O₈ (MG 722.8)

To a solution of 8.65 g (15 mmol) Z-Arg (Z₂) -OH, 3.01 g H-Phe-NH₂ · HCl, 2.45 g HOObt and 1.95 ml N-ethylmorpholine in 30 ml of dimethylformamide are added at 0 ° C with stirring 3.3 g DCC. The procedure is as in Example 1a. At the the other day the approach froze. You now rub in with semi-saturated NaHCO₃ solution, sucks off, mixed with KHSO₄ / K₂SO₄ solution, suction, washes with water and dries over P₂O₅. The substance contains approximately equimolar amounts of the dicyclohexylurea.

Yield 13.9 g.

20b. H-Arg-Phe-NH₂ · 2 HClO₄, C₁₅H₂₆Cl₂N₆O₁₀ (MG 521.33)

13.5 g of the substance obtained above (Z-Arg (Z₂) -Phe- NH₂ · dicyclohexylurea) are in 200 ml Dimethylacetamide stirred and with hydrogen and Pd / C catalytically hydrogenated. With the help of an autotitrator a pH of 5 is maintained by adding 1N HClO₄. After the hydrogenation has ended, the precipitate is filtered off with suction and concentrates the filtrate in vacuo. The arrears with Water stirs and insoluble matter is sucked off. The The filtrate is freeze-dried. The oily residue will triturated twice with diethyl ether and decanted. The oil is dried in a high vacuum.

Yield 9.5 g. The oil still contains solvents.

20c. Z-Asp (OtBu) -Arg-Phe-NH₂ · HClO₄, C₃₁H₄₄Cl₁N₇O₁₁ (MG 726.2)

To a solution of 5.9 g Z-Asp (OtBu) -OH, 9.5 g H-Arg-Phe- NH₂ · 2 HClO₄, 2.48 g HOBt and 2.37 ml N-ethylmorpholine in  50 ml of dimethylformamide are added at 0 ° C. with stirring, 4 g DCC. The batch is worked up analogously to Example 1a. The The residue is triturated with diethyl ether and suction filtered.

Yield 9 g.

20d. H-Asp (OtBu) -Arg-Phe-NH₂ · 2 HClO₄, C₂₃H₃₉Cl₂N₇O₁₃ (MW 692.5)

8.7 g of Z-Asp (OtBu) -Arg-Phe-NH₂ · HClO₄ in 150 ml Dissolved methanol and hydrogenated analogously to Example 20b. The The residue is triturated with diethyl ether and suction filtered.

Yield 7.8 g, [ α ] = -4.0 ° ( c = 1, in methanol).

20e. Z-Trp-Asp (OtBu) -Arg-Phe-NH₂-acetate, C₄₄H₅₇N₉O₁₀ (MG 872)

To a solution of 3.4 g Z-Trp-OH (10 mmol), 6.92 g H-Asp (OtBu) -Arg-Phe-NH₂ · 2 HClO₄, 1.35 g HOBt and 1.3 ml N-ethylmorpholine in 40 ml of dimethylformamide is added at 0 ° C with stirring 2.2 g DCC. The procedure is analogous to example 1a. The residue is triturated with diethyl ether and suction filtered.

Yield 7.3 g.

220 mg of the substance obtained above are 55% in 15 ml aqueous acetic acid and dissolved over a weakly basic Ion exchanger chromatographed in acetate form. You elute with 55% acetic acid. The eluate is concentrated and the residue was taken up in water and freeze-dried.

Yield 180 mg, [ α ] = -19.2 ° ( c = 1, in methanol), content of peptide base according to amino acid analysis: 75%.

21. H-Arg-Trp-Asp (OtBu) -Arg-Phe-NH₂ 21a. Z-Arg (Z₂) -Trp-Asp (OtBu) -Arg-Phe-NH₂, C₆₄H₇₉N₁₃O₁₃ (MW 1238.4)

To a solution of 576 mg (1 mmol) Z-Arg (Z₂) -OH, 879 mg H-Trp-Asp (OtBu) -Arg-Phe-NH-2 .HClO₄, 163 mg HOObt and 0.13 ml  N-ethylmorpholine in 10 ml of dimethylformamide is added Stir at 0 ° C 220 mg DCC. You let in overnight Stand at room temperature, sucks off the precipitate and constricts the filtrate in a high vacuum. The backlog is between Distributed n-pentanol and saturated NaHCO₃ solution. Here most remains undissolved and is suctioned off.

Yield 910 mg.

The n-pentanol phase is concentrated.

Yield 250 mg, [ α ] = -15.2 ° ( c = 1, in 80% aqueous acetic acid).

21b. H-Arg-Trp-Asp (OtBu) -Arg-Phe-NH₂ triacetate, C₄₆H₇₁N₁₃O₁₃ (MW 1014.15)

3 g of Z-Arg (Z₂) -Trp-Asp (OtBu) -Arg-Phe-NH₂ are in 50 ml 90% aqueous acetic acid dissolved and over Pd / C with Hydrogen catalytically hydrogenated. After finished Hydrogenation, the catalyst is suctioned off and the filtrate constricted. The residue is triturated with diethyl ether and vacuumed.

Yield 2.33 g.

960 mg of the substance obtained above are analogous to Example 18d cleaned chromatographically.

Yield 540 mg, [ α ] = -14.2 ° ( c = 1, in water), content of peptide base according to amino acid analysis: 84%.

22. Z-Asp (OtBu) -Arg-Trp-NH₂ 22a. Z-Arg (Z₂) -Trp-NH₂, C₄₁H₄₃N₇O₈ (MG 761.85)

To a solution of 17.3 g (30 mmol) of Z-Arg (Z₂) -OH, 7.2 g H-Trp-NH₂ · HCl, 4.9 g HOObt and 3.9 ml N-ethylmorpholine in 150 ml of dimethylformamide are added at 0 ° C. with stirring  6.6 g DCC. The procedure is analogous to Example 7a.

Yield 20.5 g.

For cleaning, the substance is mixed with methanol, sucks off, washes with diethyl ether and dries in vacuo.

Yield 20.3 g, mp 172-174 ° C, [ α ] = -1.4 ° ( c = 1, in dimethylformamide).

22b. H-Arg-Trp-NH₂ · 2 p-toluenesulfonic acid, C₃₁H₄₁N₇O₈S₂ (MW 703.8)

17.2 g of Z-Arg (Z₂) -Trp-NH₂ in 350 ml of methanol suspended and catalytically hydrogenated analogously to Example 2a (Instead of methanolic HCl, a 1N-p-toluenesulfonic acid Solution in methanol used). The arrears with Diethyl ether triturated and suction filtered.

Yield 12.3 g, [ α ] = + 13.3 ° ( c = 1, in methanol).

22c. Z-Asp (OtBu) -Arg-Trp-NH₂-acetate, C₃₅H₄₈N₈O₉ (MW 724.8)

To a solution of 13.5 g of H-Arg-Trp-NH₂ · 2 p-toluenesulfonic acid and 2.49 ml of N-ethylmorpholine in 50 ml of DMF are added at room temperature with stirring 8.06 g Z-Asp (OtBu) -ONSu. The mixture is left to stand at room temperature overnight. You narrow and distributes the residue between n-pentanol and Water. The organic phase is concentrated and the The residue is triturated with diethyl ether and suction filtered.

Yield 14.4 g of Z-Asp (OtBu) -Arg-Trp-NH₂ · p-toluenesulfonic acid, [ α ] = -16.4 ° ( c = 1, in methanol).

850 mg of the substance obtained above are 30% in 15 ml aqueous acetic acid and dissolved over a weakly basic Ion exchanger chromatographed in acetate form. Eluted with 30% acetic acid. The eluate is concentrated and the residue dissolved in water. The solution will be freeze-dried.  

Yield 750 mg, [ α ] = -16.6 ° ( c = 1, in methanol), content of peptide base according to amino acid analysis: 85%.

23. Z-Arg-Ile-Asp (OtBu) -Arg-Trp-NH₂ 23a. H-Asp (OtBu) -Arg-Trp-NH₂ · p-toluenesulfonic acid, C₃₉H₅₄N₈O₁₁S₂ (MG 875.05)

13.2 g of Z-Asp (OtBu) -Arg-Trp-NH₂ · p-toluenesulfonic acid analogously to Example 22b, catalytically hydrogenated.

Yield 13.04 g, [ α ] = -3.1 ° ( c = 1, in methanol).

23b. Z-Arg-Ile-Asp (OtBu) -Arg-Trp-NH₂-diacetate, C₄₉H₇₅N₁₃O₁₃ (MW 1054.2)

To a solution of 843 mg (2 mmol) Z-Arg-Ile-OH, 1.75 g H-Asp (OtBu) -Arg-Trp-NH₂ · 2 p-toluenesulfonic acid and 326 mg HOObt in 20 ml of diethylformamide is added at 0 ° C Stir 440 mg DCC. It is left overnight at room temperature stand, sucks off the precipitate and concentrates the filtrate. The residue is subjected to a 5-step Counterflow distribution between ethyl acetate and water. The Peptide is found in the water phases that are freeze-dried will.

Yield 2.01 g of Z-Arg-Ile-Asp (OtBu) -Arg-Trp-NH₂ · 2 p-toluenesulfonic acid, [ α ] = -25 ° ( c = 1, in methanol).

150 mg of the peptide obtained above are analogous to Example 22c converted into the acetate.

Yield 120 mg, [ α ] = -30.8 ° ( c = 1, in methanol), content of peptide base according to amino acid analysis: 73%.

24. H-Arg-Ile-Asp (OtBu) -Arg-Trp-NH₂-triacetate, C₄₃H₇₃N₁₃O₁₃ (MW 980.1)

1.73 g of Z-Arg-Ile-Asp (OtBu) -Arg-Trp-NH₂ · 2 p-toluenesulfonic acid become catalytic in 90% aqueous acetic acid over Pd / C  hydrated. The catalyst is suctioned off, the filtrate concentrated and the residue triturated with diethyl ether and aspirated. The substance is dissolved in water and with a weakly basic ion exchanger stirred in acetate form, until a pH of 4.4 is reached. The exchanger is filtered off and the filtrate is freeze-dried.

Yield 1.06 g.

The substance obtained above is analogous to Example 18d cleaned chromatographically.

Yield 634 mg, [ α ] = -31.9 ° ( c = 1, in water). Peptide base content according to UV spectrum: 72%.

25. Z-D-Phe-Trp-Lys-Tyr-Phe-OBzl 25a. Fmoc-Tyr (tBu) -Phe-OBzl

To a solution of 9.18 g (20 mmol) Fmoc-Tyr (tBu) -OH, 8.5 g (20 mmol) H-Phe-OBzl · p-toluenesulfonic acid, 2.8 g HOBt and 2.56 ml of N-ethylmorpholine in 50 ml of dimethylformamide 4.74 g of DCC are added at 0 ° C. with stirring. One works analogous to example 1a. From ethyl acetate / petroleum ether crystallized.

Yield 11.4 g (85%), mp. 158 ° C with decomposition, [ α ] = -34.5 ° ( c = 1, in dimethylformamide).

25b. H-Tyr (tBu) -Phe-OBzl

To a solution of 10.44 g (15 mmol) of Fmoc-Tyr (tBu) -Phe-OBzl 15.64 ml (150 mmol) are added to 100 ml of dimethylformamide Diethylamine. The mixture is stirred at room temperature for 10 minutes, concentrated and triturated the residue with petroleum ether. The Petroleum ether is decanted off and the residue in Dissolved diethyl ether. One filters off a little undissolved  and concentrates the filtrate. The residue is in a high vacuum dried.

Yield 5.2 g.

25c. Fmoc-Lys (Boc) -Tyr (tBu) -Phe-OBzl, C₅₅H₆₄N₄O₉ (MG 925.15)

To a solution of 4.38 g Fmoc-Lys (Boc) -OH, 5.2 g H-Tyr (tBu) -Phe-OBzl and 1.31 g HOObt in 50 ml Dimethylformamide is added at 0 ° C with stirring 1.97 g DCC. The procedure is analogous to example 1a. Crystallization from ethyl acetate / petroleum ether.

Yield 6.5 g.

25d. H-Lys (Boc) -Tyr (tBu) -Phe-OBzl, C₄₀H₅₄N₄O₇ (MG 702.9)

To a solution of 6.5 g (7 mmol) Fmoc-Lys (Boc) -Tyr (tBu) - Phe-OBzl in 50 ml of dimethylformamide is added to 7.3 ml Diethylamine and stirred for 10 minutes at room temperature. It is then concentrated in vacuo and the residue with Triturated diethyl ether.

Yield 5.53 g, mp. 132 ° C, [ α ] = 0 ° ( c = 1, in dimethylformamide).

25e. Z-D-Phe-Trp-OMe, C₂₉H₂₉N₃O₅ (MW 499.5)

To a solution of 7.47 g (25 mmol) of Z-D-Phe-OH, 6.37 g (25 mmol) H-Trp-OMe · HCl, 3.37 g HOBt and 3.2 ml N-ethylmorpholine in 50 ml of dimethylformamide is added at 0 ° C 5.56 g of DCC with stirring. The procedure is analogous to example 1a on. The substance crystallizes from ethyl acetate.

Yield 8.13 g, mp. 181 ° C, [ α ] = + 2.5 ° ( c = 1, in dimethylformamide).

25f. Z-D-Phe-Trp-OH, C₂₈H₂₇N₃O₅ (MW 485.5)

To a solution of 2.9 g Z-D-Phe-Trp-OMe in 20 ml Dioxane / water (4: 1) are added 6.1 ml of 1 N NaOH. You leave Stir for 2 hours at room temperature and then add 6.1 ml of 1N HCl. The solution is concentrated and the residue triturated with water. The precipitate is suctioned off and dried over P₂O₅.

Yield 2.27 g, mp 143-145 ° C, [ a ] = + 4.4 ° ( c = 1, in dimethylformamide).

25g. Z-D-Phe-Trp-Lys (Boc) -Tyr (tBu) -Phe-OBzl, C₆₈H₇₂N₇O₁₁ (MG 1170.4)

To a solution of 2.4 g (5 mmol) of Z-D-Phe-Trp-OH, 3.1 g H-Lys (Boc) -Tyr (tBu) -Phe-OBzl and 0.7 g HOBt in 30 ml Dimethylformamide is added at 0 ° C with stirring 1.13 g DCC. The procedure is analogous to example 1a. From diethyl ether / Petroleum ether is crystallized.

Yield 5.62 g (96%), mp. 194 ° C with decomposition, [ α ] = -8.5 ° ( c = 1, in dimethylformamide).

25h. Z-D-Phe-Trp-Lys-Tyr-Phe-OBzl acetate, C₆₁H₆₇N₇O₁₁ (MW 1074.2)

1.1 g (1 mmol) Z-D-Phe-Trp-Lys (Boc) -Tyr (tBu) -Phe-OBzl are implemented analogously to Example 2c. The arrears with Methyl tert-butyl ether triturated and suction filtered. The Substance is stirred with semi-saturated NaHCO₃ solution and vacuumed. The substance is dissolved in 20 ml of methanol and precipitated with 20 ml of water.

Yield 0.41 g.

The substance obtained above is converted into the acetate dissolved in 3 ml of glacial acetic acid. 70 ml of water and  freeze-dried.

Yield 0.4 g.

Amino acid analysis: Tyr 0.66; Phe 1.97; Lys 1.0; Content of Peptide base according to amino acid analysis: 75%.

26. Cyclo- (D-Phe-Trp-Lys-Tyr-Phe) 26a. H-D-Phe-Trp-Lys (Boc) -Tyr (tBu) -Phe-OH, C₅₃H₆₇N₇O₉ (MW 946.2)

4.45 g (3.8 mmol) of Z-D-Phe-Lys (Boc) -Tyr (tBu) -Phe-OBzl catalytically hydrogenated in 100 ml of 90% acetic acid over Pd / C. After the hydrogenation has ended, the catalyst is filtered off with suction and the filtrate was concentrated. The arrears with Diethyl ether triturated, suction filtered and dried.

Yield 3.25 g (90.5%).

The substance is used to remove adhering acetic acid stirred with so much 10% NaHCO₃ solution until a pH out of 6. Then the substance is sucked off with Washed water and dried over P₂O₅.

Yield 2.81 g, mp. 225-227 ° C with decomposition, [ α ] = -22.7 ° (c = 1, in dimethylformamide).

26b. Cyclo- (D-Phe-Trp-Lys (Boc) -Tyr (tBu) -Phe), C₅₃H₆₅N₇O₈ (MW 928.2)

To a solution of 0.946 g (1 mmol) H-D-Phe-Trp-Lys (Boc) - Tyr (tBu) -Phe-OH in 125 ml dimethylformamide is added 0.35 ml (2.5 mmol) of a 50% solution of ethyl methyl phosphinic anhydride in methylene chloride. Within 0.64 ml is slowly left at room temperature for 30 minutes (5 mmol) N-ethylmorpholine, dissolved in 25 ml dimethylformamide,  drop. The mixture is stirred for 3 hours at room temperature and then narrows the approach. The residue is washed with water triturated, suction filtered and dried.

Yield 0.79 g.

In order to separate non-cyclized material, 20 ml of methanol and 20 ml of water dissolved hot. After this Cooling is suctioned off.

Yield 0.33 g.

For further purification, the substance is dissolved in methylene chloride / Methanol as 95: 5 chromatographed on silica gel.

Yield 131 mg.

26c. Cyclo- (D-Phe-Trp-Lys-Tyr-Phe) acetate, C₄₆H₅₃N₇O₈ (MW 832)

105 mg of cyclo- (D-Phe-Trp-Lys (Boc) -Tyr (tBu) -Phe) treated analogously to Example 2c. It becomes analogous to example 25h worked up.

Yield 50.3 mg.

Amino acid analysis: Tyr 0.89; Phe 2.03; Lys 1.0; salary on peptide base according to amino acid analysis: 76%.

27. Z-Lys-Lys-Tyr-Phe-OMe 27a. H-Lys (Boc) -Tyr (tBu) -Phe-OMe · HCl

28 g of Z-Lys (Boc) -Tyr (tBu) -Phe-OMe (see Example 4c) analogously to Example 2a catalytically hydrogenated in methanol.

Yield 22.4 g (91.8%), mp 145-147 ° C, [ α ] = + 18.2 ° ( c = 1, in methanol).

27b. Z-Lys (Boc) -Lys (Boc) -Tyr (tBu) -Phe-OMe

To a solution of 21.9 g of H-Lys (Boc) -Tyr (tBu) -Phe-OMe · HCl, 4.46 g HOBt and 4.3 ml N-ethylmorpholine in 130 ml Dimethylformamide is added at room temperature 18.5 g Z-Lys (Boc) -OTcp and stir for 3 hours at room temperature. 400 ml of water and 35 ml are added to the mixture saturated NaHCO₃ solution. The precipitate is suctioned off. It is reprecipitated from 200 ml of ethyl acetate and 800 ml of petroleum ether.

Yield 30.4 g (93%), mp. 159-160 ° C, [ α ] = -17.5 ° ( c = 1, in methanol).

27c. Z-Lys-Lys-Tyr-Phe-OMe acetate

1.0 g of Z-Lys (Boc) -Lys (Boc) -Tyr (tBu) -Phe-OMe are dissolved in 10 ml 90% trifluoroacetic acid dissolved. Allow 1 hour Stand at room temperature. Then it is concentrated and the The residue is triturated with diethyl ether and suction filtered. The Residue is dissolved in water and weakly over a basic ion exchanger chromatographed in acetate form. The eluate is freeze-dried. Yield 736.8 mg. After chromatographic purification on silica gel:

Yield 576 mg, [ α ] = -31.1 ° ( c = 1, in water).

28. Fmoc-Arg-Tyr (tBu) -Phe-NH₂ acetate, C₄₅H₅₅N₇O₈ (MG 822)

To a solution of 3.96 g (10 mmol) of Fmoc-Arg-OH, 4.2 g (10 mmol) H-Tyr (tBu) -Phe-NH₂ · HCl (see Example 7b) and 1.4 g (10 mmol) of HOBt in 50 ml of dimethylformamide are added at 0 ° 2.2 g DCC. The mixture is left overnight at room temperature stir. Then it is concentrated and the residue with triturated saturated NaHCO₃ solution. The precipitation is suctioned off and reprecipitated from methanol / water. Yield 6.1 g. The substance is used for cleaning Chromatographed silica gel (eluent: methylene chloride / Methanol / water / glacial acetic acid like 9: 1: 0.1: 0.1.  

Yield 3.85 g, [ α ] = -7.1 ° ( c = 1, in 90% acetic acid).

29. Fmoc-D-Arg-Tyr (tBu) -Phe-NH₂-acetate, C₄₅H₅₅N₇O₈ (MG 822)

Is prepared analogously to Example 28.

[ α ] = + 8.7 ° ( c = 1, in 90% acetic acid).

30. Z-Trp-Arg-Tyr-Phe-NH₂ 30a. H-Arg-Tyr (tBu) -Phe-NH₂ acetate, C₃₀H₄₅N₇O₆ (MW 599.7)

To a solution of 3.05 g of Fmoc-Arg-Tyr (tBu) -Phe-NH₂ acetate 4.2 ml of diethylamine are added to 25 ml of dimethylformamide. Man left for 5 minutes at room temperature and concentrated in vacuo a. The residue is triturated with diethyl ether and aspirated.

Yield 2.23 g.

30b. Z-Trp-Arg-Tyr (tBu) -Phe-NH₂ acetate, C₄₉H₆₁N₉O₉ (MW 920.1)

To a solution of 2.15 g (3.59 mmol) H-Arg-Tyr (tBu) - Phe-NH₂ acetate, 0.56 g HOBt and 0.64 g (3.59 mmol) Pyridinium perchlorate in 25 ml of dimethylformamide 2.17 g of Z-Trp-OTcp at room temperature. One leaves over Stand at room temperature overnight and constrict the next day. The residue is divided between ethyl acetate and saturated NaHCO₃ solution. The ethyl acetate phase will dried over Na₂SO₄ and concentrated. The backlog will triturated with diethyl ether. Yield 2.61 g. For cleaning is chromatographed on silica gel as in experiment 28a.

Yield 1 g.

30c. Z-Trp-Arg-Tyr-Phe-NH₂ acetate, C₄₅H₅₃N₉O₉ (MG 864)

700 mg Z-Trp-Arg-Tyr (tBu) -Phe-NH₂ are analogous to Example 2c treated.  

Yield 0.38 g, [ α ] = -5.7 ° ( c = 1, in 90% acetic acid).

31. Z-Trp-D-Arg-Tyr-Phe-NH₂ acetate, C₄₅H₅₃N₃O₃ (MW 864)

Is prepared analogously to Example 30.

[ α ] = + 6.0 ° ( c = 1, in 90% acetic acid).

32. Z-Trp-D-Lys-Tyr-Phe-NH₂, C₄₃H₄₉N₇O₇ (MW 775.9)

Wrd produced analogously to Example 8.

[ α ] = + 6.5 ° ( c = 1, in 90% acetic acid).

33. Indole-3-butyryl-Trp-Lys-Tyr-Phe-NH₂ 33a. Indole-3-butyryl-Trp-Lys (Boc) -Tyr (tBu) -Phe-NH₂, C₅₆H₇₀N₈O₈ (MG 983.24)

To a solution of 509 mg indole-3-butyric acid, 2.1 g H-Trp-Lys (Boc) -Tyr (tBu) -Phe-NH₂ · HCl (see Example 8c, Catalytic hydrogenation analogous to Example 2a), 337 mg HOBt and 0.325 ml of N-ethylmorpholine in 20 ml of dimethylformamide one at 0 ° C 550 mg DCC. Work up as in Example 1a. The residue is triturated with diethyl ether and suction filtered.

Yield 2.15 g, mp 148-155 ° C, [ α ] = -15.4 ° ( c = 1, in methanol).

33b. Indole-3-butyryl-Trp-Lys-Tyr-Phe-NH₂ acetate, C₄₉H₅₈N₈O₈ (MW 887.05)

1.97 g of indole-3-butyryl-Trp-Lys (Boc) -Tyr (tBu) -Phe-NH₂ implemented analogously to Example 2c. Yield 1.29 g. Cleaned is by column chromatography on silica gel with Methylene chloride / methanol / water / glacial acetic acid as 7: 2: 0.3: 0.15.

Yield 397 mg, [ α ] = -2.7 ° ( c = 1, in 10% acetic acid).

34. Indole-3-butyryl-Lys-Tyr-Phe-NH₂ 34a. Indole-3-butyryl-Lys (Boc) -Tyr (tBu) -Phe-NH₂, C₄₅H₆₀N₆O₇ (MW 797.02)

To a solution of 1.95 g (3 mmol) H-Lys (Boc) -Tyr (tBu) - Phe-NH₂ · HCl, 610 mg indole-3-butyric acid, 405 mg HOBt and 0.39 ml of N-ethylmorpholine in 20 ml of dimethylformamide are added at 0 ° C 660 mg DCC. Work up as in Example 1a. The Substance is precipitated from ethyl acetate / petroleum ether.

Yield 2.4 g, mp. 168 °, [ α ] = -21.4 ° ( c = 1, in methanol).

34b. Indole-3-butyryl-Lys-Tyr-Phe-NH₂-acetate, C₃₈H₄₈N₆O₇ (MW 700.85)

2.15 g of indole-3-butyryl-Lys (Boc) -Tyr (tBu) -Phe-NH₂ implemented and cleaned as in Example 33b.

Yield 402 mg, [ α ] = -22.9 ° ( c = 1, in water).

35. Indole-2-carbonyl-Lys-Tyr-Phe-NH₂ 35a. Indole-2-carbonyl-Lys (Boc) -Tyr (tBu) -Phe-NH₂, C₄₂H₅₄N₆O₇ (MW 754.9)

485 mg (3 mmol) indole-2-carboxylic acid are analogous to the example 34a with 1.95 g (3 mmol) of H-Lys (Boc) -Tyr (tBu) -Phe-NH₂ · HCl implemented.

Yield 2.25 g, mp 228-230 ° C, [ α ] = -56.7 ° ( c = 1, in methanol).

35b. Indole-2-carbonyl-Lys-Tyr-Phe-NH₂-acetate, C₃₅H₄₂N₆O₇ (MW 658.7)

2 g of indole-2-carbonyl-Lys (Boc) -Tyr (tBu) -Phe-NH₂ be implemented analogously to Example 33b.  

Yield 1.55 g, [ α ] = -44.8 ° ( c = 1, in methanol).

36. H-Trp-Arg-Trp-Gly-Trp-Arg-Trp-Gly-OH 36a. Fmoc-Trp-Gly-OtBu, C₃₂H₃₃N₃O₅ (MG 539.64)

To a solution of 21.3 g (50 mmol) of Fmoc-Trp-OH, 16.7 g (50 mmol)) H-Gly-OtBu · HCl and 7 g HOBt in 100 ml DMF are added at 0 ° C 11.33 g DCC. Work up as in Example 1a. The Compound crystallizes from diethyl ether.

Yield 17.26 g, mp. 141 ° C, [ α ] = -26.6 ° ( c = 1, in dimethylformamide).

36b. Fmoc-Trp-Arg-OH, C₃₂H₃₄N₆O₅ (MW 582.67)

To a suspension of 6.09 g (35 mmol) arginine and 6.25 g (35 mmol) pyridinium perchlorate in 75 ml dimethylformamide 19.98 g (37 mmol) of Fmoc-Trp-OObt are added. You let stir until everything is solved. Then you concentrate and stir the Residue with NaHCO₃ solution (pH of the suspension should be about 6 to 7). The precipitate is suctioned off and dried.

Yield 17.65 g, mp. 185 ° C (dec.), [ A ] = -24.2 ° ( c = 1, in dimethylformamide).

36c. Fmoc-Trp-Arg-Trp-Gly-OtBu, C₄₉H₅₅N₉₃O₇ (MW 882.05)

To a solution of 5.4 g (10 mmol) Fmoc-Trp-Gly-OtBu in 25 ml of dimethylformamide are added to 10.3 ml (100 mmol) Diethylamine. The mixture is left to stand at room temperature for 10 minutes and constricts. The residue is washed several times with petroleum ether digested. The residue is dissolved in diethyl ether, from Insoluble filtered and concentrated. Yield 4.2 g light Oil (contaminated).  

To a solution of 4.2 g of oil obtained, 5.82 g (10 mmol)) Fmoc-Trp-Arg-OH and 1.6 g HOObt in 30 ml Dimethylformamide is added at 0 ° C, 2.2 g DCC. One leaves over Standing at room temperature overnight, sucks off the precipitate, constricts and divides between ethyl acetate and saturated NaHCO₃ solution. The ethyl acetate solution is mixed with water shaken out and concentrated. The arrears with Diethyl ether triturated, suction filtered and dried.

Yield 8.48 g, mp 145-152 ° C, [ α ] = -22.4 ° ( c = 1, in dimethylformamide).

36d. Fmoc-Trp-Arg-Trp-Gly-OH

2.64 g (3 mmol) of Fmoc-Trp-Arg-Trp-Gly-OtBu become analogous Example 2c implemented. The residue is treated with methyl tert. butyl ether triturated and suction filtered. After drying the substance is stirred with NaHCO₃ solution (pH of the suspension 6 to 7). The substance is suctioned off and dried.

Yield 1.82 g, mp. 190-192 ° C (dec.), [ Α ] = -43.7 ° ( c = 1, in dimethylformamide).

36e. Fmoc-Trp-Arg-Trp-Gly-Trp-Arg-Trp-Gly-OtBu

A solution of 2.64 g (3 mmol) Fmoc-Trp-Arg-Trp-Gly-OtBu in 20 ml of DMF are mixed with 3.3 ml of diethylamine. To The mixture is concentrated for 5 minutes at room temperature and the The residue is triturated with diethyl ether and suction filtered. Yield 2.04 g H-Trp-Arg-Trp-Gly-OtBu. To a Solution of 1.81 g (2.2 mmol) of Fmoc-Trp-Arg-Trp-Gly-OH, 1.45 g (2.2 mmol) H-Trp-Arg-Trp-Gly-OtBu, 0.9 g (2.2 mmol) Pyridinium perchlorate and 0.36 g (2.2 mmol) HOB in 25 ml Dimethylformamide is added at 0 ° C to 0.51 g (2.5 mmol) DCC. It is left to stand at room temperature overnight, the is sucked Precipitation and the filtrate is concentrated. The residue is triturated with saturated NaHCO₃ solution, suction filtered and washed with water.  

Yield 3.3 g, mp 204-205 ° C (dec.), [ Α ] = -27.6 ° ( c = 1, in dimethylformamide).

36f. H-Trp-Arg-Trp-Gly-Trp-Arg-Trp-Gly-OH-diacetate, C₆₄H₈₀N₁₈O₁₃ (MG 1309.5)

To a solution of 2.93 g (2 mmol) of Fmoc-Trp-Arg-Trp-Gly- Trp-Arg-Trp-Gly-OH in 20 ml dimethylformamide are added 2.1 ml (20 mmol) diethylamine. The mixture is left at room temperature for 5 minutes stand and constrict. The residue is mixed with diethyl ether rubbed. Yield 2.7 g H-Trp-Arg-Trp-Gly-Trp-Arg-Trp-Gly-OH.

1 g of the substance obtained above are reacted analogously to Example 2c. Purification of the substance on an alkylated dextran gel. Yield 210 mg. Characterization by UV: characteristic band for Trp at 280 nm; Amino acid analysis in 6N HCl: 2.00 Gly, 2.04 Arg; Peptide base content: 70%, [ α ] = + 1 ° ( c = 1, in 90% acetic acid).

37. cyclo- (Trp-Arg-Trp-Gly-Trp-Arg-Trp-Gly) diacetate, C₆₄H₇₈N₁₈O₁₄ (MG 1291.4)

1.6 g of H-Trp-Arg-Trp-Gly-Trp-Arg-Trp-Gly-OtBu are added Room temperature in a mixture of 16 ml 90% Trifluoroacetic acid and 1.4 ml of 1,2-dimercaptoethane dissolved. After 1 hour, the mixture is concentrated and the residue is tert-butyl ether triturated and suction filtered. Yield 1.42 g. The dry material obtained in this way is treated with a saturated NaHCO₃ solution stirred (pH of the suspension at 7), vacuumed and dried. Yield 0.92 g H-Trp-Arg-Trp- Gly-Trp-Arg-Trp-Gly-OH-ditrifluoroacetate.

To a solution of the substance obtained from above and 0.185 g HOBt in 6 ml dimethylformamide is added at 0 ° C 0.2 g DCC. The mixture is left to stand at room temperature overnight. The next day  the precipitate is filtered off, the filtrate is concentrated and the residue triturated with NaHCO₃ solution, suction filtered and dried. Yield 1.05 g. You now solve in a mixture from 10 ml of glacial acetic acid and 40 ml of water and chromatographed via a weakly basic ion exchanger in acetate Shape. The eluate is concentrated. Yield 0.83 g. To Purification is via an alkylated dextran gel in one Mixture of methanol / water / acetic acid such as 50: 50: 3 chromatographed. Yield 101 mg. characterization by UV: characteristic band for Trp at 280 nm; Amino acid analysis in 6 N HCl: 2.00 Gly, 1.9 Arg; salary on peptide base: 82%.

38. Z-Trp-D-Lys-Tyr-Ile-Lys-Pro-OBzl 38a. Fmoc-Ile-Lys (Boc) -OH, C₃₂H₄₃N₃O₇ (MG 581.7)

To a suspension of 19.76 g H-Lys (Boc) -OH in 120 ml Dimethylformamide is added to 40 g of Fmoc-Ile-OObt. You stir until everything is solved and then narrows down. The backlog will between ethyl acetate and semi-saturated NaHCO₃ solution distributed. The ethyl acetate phase is repeated 2 times with the NaHCO₃ solution, 2 times with KHSO₄ solution and 1 time with water shaken out, dried over Na₂SO₄ and concentrated. The Substance is still contaminated with Fmoc-Ile-OH and will therefore 5 minutes with 100 ml of methyl tert-butyl ether heated and vacuumed in the steam bath.

Yield 28.85 g, mp 162-164 °, [ α ] = -15.5 ° ( c = 1, in methanol).

38b. Fmoc-Ile-Lys (Boc) -Pro-OBzl, C₄₄H₅₆N₄O₈ (MG 768.9)

To a solution of 28 g Fmoc-Ile-Lys (Boc) -OH, 11.6 g H-Pro-OBzl · HCl, 7.8 g HOObt and 6.3 ml N-ethylmorpholine 10.6 g of DCC are added to 250 ml of dimethylformamide at 0 ° C. Work up as in Example 1a. It leaves an oil  that over silica gel in methylene chloride / methanol / water how 9: 1: 0.1 is chromatographed.

Yield: 30.4 g of yellowish oil.

38c. H-Ile-Lys (Boc) -Pro-OBzl, C₂₉H₄₆N₄O₆ (MG 546.7)

To a solution of 28.86 g Fmoc-Ile-Lys (Boc) -Pro-OBzl in 200 ml of dimethylformamide are added to 39 ml of diethylamine. Man leave for 10 minutes at room temperature, concentrate and chromatograph the residue on silica gel in Methylene chloride and later in methylene chloride / methanol like 9: 1.

Yield 18.4 g of yellowish oil.

38d. Fmoc-D-Lys (Boc) -Tyr (tBu) -OH, C₃₉H₄₉N₃O₈ (MW 687.8)

To a suspension of 11.6 g H-Tyr (tBu) -OH in 150 ml Dimethylformamide is added to 30 g of Fmoc-D-Lys (Boc) -OObt. Man proceeds as in Example 38a. For cleaning it will Peptide twice from methyl tert-butyl ether / petroleum ether like.

Yield 16.45 g, mp 92-94 ° C, [ α ] = -12.8 ° ( c = 1, in methanol).

38e. Fmoc-D-Lys (Boc) -Tyr (tBu) -Ile-Lys (Boc) -Pro-OBzl, C₆₈H₉₃N₇O₁₃ (MG 1216.5)

To a solution of 16.1 g of Fmoc-D-Lys (Boc) -Tyr (tBu) -OH, 12.8 g H-Ile-Lys (Boc) -Pro-OBzl and 3.8 g HOObt in 150 ml 5.15 g of DCC are added to dimethylformamide. After about 4 hours Reaction time at room temperature becomes the precipitate suction filtered and the filtrate concentrated. The backlog will successively with ethyl acetate, semi-saturated NaHCO₃ solution and rubbed and sucked off water. Made from isopropanol / Petroleum ether is knocked over.

Yield 18.9 g, mp 147-149 ° C, [ α ] = -31.7 ° ( c = 1, in methanol).

38f. H-D-Lys (Boc) -Tyr (tBu) -Ile-Lys (Boc) -Pro-OBzl, C₅₃H₈₃N₇O₁₁ (MG 994.3)

To a solution of 18 g (14.8 mmol) Fmoc-D-Lys (Boc) - Tyr (tBu) -Ile-Lys (Boc) -Pro-OBzl in 150 ml dimethylformamide 15.4 ml of diethylamine are added at room temperature. To It is concentrated for 20 minutes and the residue on silica gel chromatographed. The impurities are with Methylene chloride / methanol like 98: 2 removed and the substance eluted with methylene chloride / methanol as 90:10.

Yield 10.4 g of yellowish oil.

38g. Z-Trp-D-Lys (Boc) -Tyr (tBu) -Ile-Lys (Boc) -Pro-OBzl, C₇₂H₉₃N₉O₁₄ (MG 1308.6)

To a solution of 10.4 g H-D-Lys (Boc) -Tyr (tBu) -Ile- Lys (Boc) -Pro-OBzl, 3.54 g Z-Trp-OH and 1.41 g HOBt in 150 ml of dimethylformamide are added at 0 ° C to 2.3 g of DCC. Work up as in Example 1a. The backlog is out Diethyl ether / petroleum ether precipitated.

Yield 12.65 g, mp 111-113 ° C, [ α ] = -26.4 ° ( c = 1, in methanol).

38h. Z-Trp-D-Lys-Tyr-Ile-Lys-Pro-OBzl-diacetate, C₆₂H₈₃N₉O₁₄ (MW 1178.4)

1.31 g Z-Trp-D-Lys (Boc) -Tyr (tBu) -Ile-Lys (Boc) -Pro-OBzl are implemented analogously to Example 2c. For cleaning overturned from isopropanol / petroleum ether.

Yield 892 mg, [ α ] = -31.6 ° ( c = 1, in water).

39. cyclo- (Trp-D-Lys-Tyr-Ile-Lys-Pro) 39a. H-Trp-D-Lys (Boc) -Tyr (tBu) -Ile-Lys (Boc) -Pro-OH, C₅₇H₈₇N₉O₁₂ (MG 1090.4)

10.5 g Z-Trp-D-Lys (Boc) -Tyr (tBu) -Ile-Lys (Boc) -Pro-OBzl are catalytically hydrogenated in 90% acetic acid. The  The residue is distributed between water and ethyl acetate. The ethyl acetate phase is dried over Na₂SO₄ and constricted. The residue is triturated with diethyl ether and vacuumed. Yield 7.0 g. Adhesive for removal Acetic acid is stirred with NaHCO₃ solution at pH 7.

Yield 7.0 g, mp 174-176 ° C, [ α ] = -18.9 ° ( c = 1, in methanol).

39b. cyclo- (Trp-D-Lys (Boc) -Tyr (tBu) -Ile-Lys (Boc) -Pro), C₅₇H₈₅N₉O₁₁ (MG 1072.4)

To a solution of 1.09 g H-Trp-D-Lys (Boc) -Tyr (tBu) -Ile- Lys (Boc) -Pro-OH and 270 mg HOBt in 200 ml dimethylformamide give a solution of 440 mg DCC in 100 ml Dimethylformamide. After two days at room temperature there 135 mg of DCC are added and the mixture is left for another 2 days React to room temperature. Then it is concentrated and the Rubbed residue with water and NaHCO₃ solution and aspirated. The substance is in silica gel Methylene chloride / methanol / acetic acid / water as 150: 8: 1: 1 chromatographed.

Yield 500 mg.

39c. cyclo- (Trp-D-Lys-Tyr-Ile-Lys-Pro) diacetate, C₄₇H₆₉N₉O₁₁ (MW 936.12)

500 mg cyclo- (Trp-D-Lys (Boc) -Tyr (tBu) -Ile-Lys-Pro) implemented analogously to Example 2c. Yield 234 mg. For cleaning chromatographed on an alkylated dextran gel. Yield 109.5 mg. Amino acid analysis (hydrolysis in 6 N HCl, 24 hours at 120 ° C): 0.98 Pro, 1.01 Ile, 2.00 Lys, 1.04 Tyr; UV spectrum: characteristic band at 280 nm for Trp.  

40. Z-Trp-Lys-Tyr-benzylamide 40a. Z-Trp-Lys (Boc) -OH, C₃₀H ⁰⁸⁷¹² ⁰⁰⁰⁷⁰ ⁵⁵² ⁰⁰¹⁰⁰⁰²⁸⁰⁰⁰⁰⁰00200012000285910860100040 0002003813989 00004 08593½N₄O₇ (MG 566.7)

To a suspension of 18.45 g (75 mmol) H-Lys (Boc) -OH 38.64 g (80 mmol) are added to 120 ml of dimethylformamide Fmoc-Trp-OObt. You stir until everything is solved. After that concentrated, the residue dissolved in ethyl acetate and with NaHCO₃ solution, KHSO₄ solution and water shaken out. The The ethyl acetate phase is dried over Na₂SO₄ and concentrated. The residue is chromatographed on silica gel. To remove impurities, first use Methylene chloride / acetone as 9: 1 and for elution of the substance rinsed with methylene chloride / methanol as 95: 5.

Yield 30 g, mp 95-97 ° C, [ α ] = -16.4 ° ( c = 1, in dimethylformamide).

40b. Z-Trp-Lys (Boc) -Tyr-OMe, C₄₀H₄₉N₅O₉ (MG 743.9)

To a solution of 16.68 g (30 mmol) of Z-Trp-Lys (Boc) -OH, 6.95 g (30 mmol) of H-Tyr-OMe · HCl and 4.92 g (30 mmol) of HOObt in 120 ml of dimethylformamide, 3.84 ml are added at 0 ° C N-ethylmorpholine and 6.8 g DCC. Refurbishment as with Example 1a. The residue is triturated with petroleum ether and dried in a high vacuum.

Yield 25.3 g of amorphous substance.

40c. Z-Trp-Lys (Boc) -Tyr-OH, C₃₉H₄₇N₅O₉ (MW 729.8)

To a solution of 25 g Z-Trp-Lys (Boc) -Tyr-OMe in 120 ml Dioxane is added to 70 ml of water and 35 ml of 1N NaOH. After 6 hours Stirring at room temperature, the dioxane is distilled off and Acidified to pH 2 with KHSO₄ solution. The leaving Oil is shaken into ethyl acetate. The ethyl acetate phase is dried over Na₂SO₄ and concentrated. The resulting Oil is poured over silica gel in methylene chloride / methanol / water / Acetic acid such as 110: 10: 1: 1 chromatographed.

Yield 13.3 g of amorphous substance, mp. 142-150 ° C (dec.), [ Α ] = -12.2 ° ( c = 1, in dimethylformamide).

40d. Z-Trp-Lys (Boc) -Tyr-benzylamide, C₄₆H₅₄N₆O₈ (MG 819)

To a solution of 2.19 g (3 mmol) of Z-Trp-Lys (Boc) -Tyr-OH and 0.492 g HOObt in 20 ml dimethylformamide are added 0.33 ml (3 mmol) benzylamine and at 0 ° C 0.68 g DCC. Refurbishment as in Example 1a. It is reprecipitated from methanol / water.

Yield 1.5 g, mp. 218-221 ° C (dec.), [ Α ] = -25.5 ° ( c = 1, in dimethylformamide).

40e. Z-Trp-Lys-Tyr-benzylamide acetate, C₄₃H₅₀N₆O₈ (778.9)

1.3 g of Z-Trp-Lys (Boc) -Tyr-benzylamide are analogous to Example 2c implemented. Yield 0.92 g. Purification by chromatography on silica gel in methylene chloride / methanol / water / acetic acid like 130: 30: 4: 2.

Yield 335 mg, [ a ] = -6.1 ° ( c = 1, in 90% acetic acid).

41. Z-Trp-Lys-Tyr-phenethylamide 41a. Z-Trp-Lys (Boc) -Tyr-phenethylamide, C₄₇H₅₆N₆O₈ (MW 833.0)

Preparation analogous to example 40d with 0.38 ml (3 mmol) Phenethylamine.

Yield 1.48 g, mp. 215-218 ° C (dec.), [ Α ] = -23.6 ° ( c = 1, in dimethylformamide).

41b. Z-Trp-Lys-Tyr-phenethylamide acetate, C₄₄H₅₂N₆O₈ (MW 792.9)

1.3 g of Z-Trp-Lys (Boc) -Tyr-phenethylamide are analogous Example 40e implemented and cleaned.

Yield 438 mg, [ a ] = -3.9 ° ( c = 1, in 90% acetic acid).

42. Z-Trp-Lys-Tyr-Phe-ol 42a. Z-Trp-Lys (Boc) -Tyr-Phe-ol, C₄₈H₅₈N₆O₉ (MW 863.0)

Preparation analogous to example 40d with 0.95 g (3 mmol) of H-phenol · HOObt.

Yield 1.47 g, mp. 194-196 ° C (dec.), [ Α ] = -44.9 ° ( c = 1, in dimethylformamide).

42b. Z-Trp-Lys-Tyr-Phe-ol acetate, C₄₅H₅₄N₆O₉ (MW 822.9)

1.3 g of Z-Trp-Lys (Boc) -Tyr-Phe-ol are analogous to Example 40e implemented and cleaned.

Yield 423 mg, [ α ] = -18.8 ° ( c = 1, in 90% acetic acid).

43. Z-Trp-Lys-Tic-NH₂ 43a. Z-Trp-Lys (Boc) -Tic-NH₂, C₄₀H₄₄N₆O₇ (MW 720.8)

To a solution of 0.99 g (1.75 mmol) Z-Trp-Lys (Boc) -OH, 0.6 g H-Tic-NH₂-tosylate and 0.29 g HOObt in 10 ml Dimethylformamide is added at 0 ° C to 0.23 ml of N-ethylmorpholine and 0.38 g DCC. Working up analogously to Example 1a. The Substance is reprecipitated from ethyl acetate / petroleum ether.

Yield 1.05 g, mp 160-175 ° C (dec.), [ Α ] = -14.3 ° ( c = 1, in dimethylformamide).

43b. Z-Trp-Lys-Tic-NH₂ acetate, C₃₇H₄₄N₆O₇ (684.8)

0.85 g of Z-Trp-Lys (Boc) -Tic-NH₂ are analogous to Example 40e implemented and cleaned.

Yield 195 mg, [ α ] = -24.1 ° ( c = 1, in 90% acetic acid).

44. Z-D-Phe-D-Tyr-D-Lys-D-Trp-NH₂ 44a. Z-D-Lys (Boc) -D-Trp-NH₂, C₃₀H₃₉N₅O₆ (MW 565.7)

To a solution of 4.3 g (18 mmol) H-D-Trp-NH₂ · HCl, 2.52 g (18 mmol) HOBt and 7.22 g Z-D-Lys (Boc) -OH in 75 ml Dimethylformamide is added at 0 ° C to 2.3 ml of N-ethylmorpholine and 3.91 g DCC. Working up is carried out analogously to Example 1a. The The residue is triturated with methyl tert-butyl ether.

Yield 9.05 g, mp 200-205 ° C (dec.), [ Α ] = 11.2 ° ( c = 1, in dimethylformamide).

44b. H-D-Lys (Boc) -D-Trp-NH₂, C₂₂H₃₄Cl₁N₅O₄ (MW 468.0)

8.75 g of Z-D-Lys (Boc) -D-Trp-NH₂ are analogous to Example 2a catalytically hydrogenated.

Yield 7.25 g, mp 170-185 ° C (dec.), [ Α ] = -1.7 ° ( c = 1, in dimethylformamide).

44c. Z-D-Tyr (tBu) -D-Lys (Boc) -D-Trp-NH₂, C₄₃H₅₆N₆O₈ (MW 784.9)

To a solution of 7.02 g (15 mmol) of H-D-Lys (Boc) -D-Trp- NH₂ · HCl, 6 g (16 mmol) Z-D-Tyr (tBu) -OH and 2.1 g HOBt in 75 ml of dimethylformamide are added at 0 ° C to 1.92 ml (15 mmol) N-ethylmorpholine and 3.5 g DCC. Refurbishment as with Example 1a. Rub the residue with diethyl ether.

Yield 8.54 g, mp. 178-185 ° C (dec.), [ Α ] = 10.8 ° ( c = 1, in dimethylformamide).

44d. H-D-Tyr (tBu) -D-Lys (Boc) -D-Trp-NH₂ · HCl, C₃₅H₅₁Cl₁N₆O₆ (MG 687.3)

8.3 g (10.5 mmol) of Z-D-Tyr (tBu) -D-Lys (Boc) -D-Trp-NH₂ analogously to Example 2a catalytically hydrogenated.

Yield 7.14 g, mp. 190-192 ° C (dec.), [ Α ] = -11.2 ° ( c = 1, in dimethylformamide).

44e. Z-D-Phe-D-Tyr (tBu) -D-Lys (Boc) -D-Trp-NH₂, C₅₂H₆₅N₇O₉ (MW 932.1)

To a solution of 2.74 g (4 mmol) H-D-Tyr (tBu) -D-Lys (Boc) - D-Trp-NH₂ · HCl, 1.21 g (4 mmol) Z-D-Phe-OH and 0.66 g HOObt 0.52 ml are added to 25 ml of dimethylformamide at 0 ° C N-ethylmorpholine and 1.1 g DCC. Refurbishment as with Example 1a.

Yield 2.77 g, mp. 179-185 ° C (dec.), [ Α ] = 20.1 ° ( c = 1, in dimethylformamide).

44f. Z-D-Phe-D-Tyr-D-Lys-D-Trp-NH₂-acetate, C₄₅H₅₃N₇O₉, (MG 836)

1 g of Z-D-Phe-D-Tyr (tBu) -D-Lys (Boc) -D-Trp-NH₂ are analogous Example 2c implemented.

Yield 434.1 mg, [ α ] = 4.9 ° ( c = 1, in 90% acetic acid).

used abbreviations

The abbreviations used for amino acids correspond the three-letter code common in peptide chemistry, such as he z. B. in Europ. J. Biochem. 138 9 (1984). Some other abbreviations are listed below:

Ac-Acetyl Boc-tert.-Butyl-oxycarbonyl-DCCDicyclohexylcarbodiimid DMFDimethylformamid Et-Ethyl Fmoc-9-Fluorenyl-methyl-oxycarbonyl-HOBt1-Hydroxy-Benzotriazol HOObt3-Hydroxy-4-oxo-3.4-dihydro-1.2.3-benzotriazole olein reduced α- carboxyl group of a C-terminal amino acid residue -OMeMethyl-ester -ONSuN-hydroxysuccinimide-ester -OtButert.-Butyl-ester -OTcp2.3.4-Trichlorphenylester -StButert.-Butylmercapto -tButert.-Butyl TicTetrahydroisoch -carboxylic acid Z-benzyl-oxycarbonyl-

Claims (9)

1. Peptide whose C-terminal α- carboxyl group is protected by an ester function or an amide function, the general formula I LBA (I) in which L is absent or a lipophilic residue, leg basic amino acid and A aromatic amino acid, and its physiologically acceptable salts. 2. A peptide of the general formula I as claimed in claim 1, in which L is an N-terminally protected lipophilic amino acid protected by lipophilic acyl or peptide residues in the L or D configuration, which is acyl-like bonded to the α- amino group of the basic amino acid, or represents a lipophilic acyl radical or a (C₁-C₁₇) alkylcarbonyl radical, and its physiologically tolerable salts. 3. peptide of the general formula I according to claim 1, wherein LIsoleucin, leucine, methionine, S-protected cysteine, valine; phenylalanine, tryptophan or naphthylalanine optionally substituted by halogen, (C₁-C₄) alkyl or (C₁-C₄) alkoxy; on the hydroxyl group substituted by (C₁-C₄) alkyl tyrosine, serine or threonine; histidine substituted on the N of the imidazole ring by (C₁-C₄) alkyl; aspartic acid, glutamic acid or aminoadipic acid esterified in the side chain by (C₁-C₄) -alkyl or amidated by lipophilic amines; or on the ω- amino group with (C₁-C L) alkylcarbonyl substituted or unsubstituted lysine or ornithine; lipophilic residues or (C₁-C₁₇) alkylcarbonyl; Optionally alkylated on the guanidino group (C₁-C₆) arginine or homoarginine, lysine, ornithine, hydroxylysine or citrulline in the D or L configuration and optionally in the aromatic system by 1 to 5 identical or different radicals from the halogen series, (C₁ -C₄) alkyl or (C₁-C₄) alkoxy substituted phenylalanine, tryptophan, naphthylalanine, thienylalanine or tetrahydro-isoquinoline-3-carboxylic acid; or tyrosine optionally substituted on the hydroxy group by (C₁-C₄) alkyl in the L or D configuration, the carboxyl groups of which are substituted by (C₁-C₄) alkyl or (C₅-C₁₄) aryl- (C₁-C₄) - alkyl esters, by the amide group, by primary (C₁-C₄) alkyl, (C₅-C₁₄) aryl (C₁-C₄) alkylamides and peptide residues consisting of up to 4 naturally occurring amino acids, are protected, and its physiologically acceptable salts. 4. Process for the preparation of a peptide of formula I. according to one or more of claims 1 to 3, characterized in that a fragment with terminal carboxyl group or its reactive Derivative with a corresponding fragment with free Amino group couples, possibly to protect others functional groups (a) temporarily introduced Protective group (s) split off and the thus obtained Compound in their physiologically acceptable salt transferred. 5. Use of a peptide of formula I according to one or several of claims 1 to 3 as a remedy. 6. Use of a peptide of formula I according to one or several of claims 1 to 3 for treatment inflammatory, allergic, rheumatic and autoimmune diseases.   7. Use of a peptide of formula I according to one or several of claims 1 to 3 for the treatment of Blood disorders, skin disorders as well Rejection after transplantation. 8. Pharmaceutical composition containing a peptide of Formula I according to one of claims 1 to 3. 9. Process for the manufacture of a pharmaceutical Means according to claim 8, characterized in that a peptide of the general formula I or its physiologically acceptable salt together with a physiologically acceptable carrier and optionally further additives, auxiliaries and / or preservatives into a suitable dosage form.