DE2308362A1 - PSYCHOPHARMACOLOGICALLY ACTIVE PEPTIDES AND PEPTIDE DERIVATIVES - Google Patents

Description

AKZO NV
IJssellaan 82, Arnhem, The Netherlands

concerning
Psychopharmacologically active peptides and peptide derivatives

From European Journal of Pharmacology 2, 14 (1967) it is known that certain peptide fragments of natural adrenocorticotrop-.en hormones (ACTH) the extinction of the learned Prevent an evasive response (canditicned avoidance response). Especially the peptide with the amino acid sequence 1-10 of ACTH proved effective in this regard. In addition, it has been shown that the first three amino acids (Ser-Tyr-Ser) could even be omitted entirely without a substantial loss of activity. The article ends with the conclusion, that a peptide with the amino acid sequence 4-10 of "ACTH" is the shortest peptide and possibly the sole sequence for said activity. '

The peptide with the amino acid sequence 4-10 of ACTH does not only show the psychopharmacological ' Activity, but also a slight MSH activity, as seen at

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such fragments of ACTH is common. Although the effect of a low dose of an MSH-active peptide in humans not yet known, studies have been made to find peptides that have the same psychopharmacological Have activity, but no or at least reduced MSH activity.

First of all, it has been shown that the 4-10 sequence of ACTH is not the key sequence for this psychopharmacological Activity, but a much smaller peptide with the amino acid sequence 4-6 of ACTH.

It showed that the peptide H-L-Met-L-Glu-L-His-OH is the is the shortest peptide that prevents the genetic evacuation reaction from being extinguished to the same extent as the well-known 4-10 ACTH, while the tripeptide has no detectable MSH activity.

It was found that an extension of the chain on the side with the terminal carbon atom of the tripeptide with -Phe-OH (4-7 ACTH), -Phe-Lys (or Arg) -0H (4-Θ ACTH), -Phe-Lys (or Arg) -Trp-0H (4-9 ACTH) or -Phe- Lys (or Arg) -Trp-Gly-OH (4-10 ACTH) did not lead to a significant increase in activity.

It was also observed that the amino acid residues

with the terminal carbon atom of the 4-7, 4-8 and 4-9 ACTH peptides

, through groups

could be replaced / which differ from the amino acid residues mentioned with terminal carbon atoms differ mainly by the absence of the carboxyl group there was a marked decrease in activity.

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For example, the amino acid residue Phe could be terminated with .C atom in the tetrapeptide 4-7 ACTH (Met-GIu-His-Phe) replaced are replaced by a (N-phenylalkyl) amino group of the general formula

- N - Alk - //

in which Alk denotes a branched or unbranched alkylene group with 1-6 carbon atoms and E ₁ denotes a hydrogen or halogen atom or a hydroxy or alkyl or alkoxy group with 1-4 carbon atoms.

The amino acid residue with a terminal C-atom Lys or Arg in the pentapeptide 4-8 ACTH (Met-Glu-His-Phe-Lys or Arg) could be replaced by an (N-aminoalkyl) amino group of the general formula "

- H - Alk - Ni '

in which Alk is a branched or unbranched alkylene group having 2-6 carbon atoms, Rp is ^a hydrogen atom or a lower alkyl group and IU is a hydrogen atom, a lower alkyl or an amidine group.

The amino acid residue Trp with a terminal carbon atom in the Hexapeptide 4-9 ACTH (Met-Glu-His-Phe-Lys (or Arg) -Trp could be replaced by an (N-3-indolylalkyl) amino group of the general formula

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in the alk a branched or non-branched alkylene group means with 1-6 carbon atoms. The latter modification appeared to be particularly cheap.

Surprisingly, it has now been shown that the amino acid terminated with N-atom (L-Met) in the above-mentioned peptides and peptide derivatives believed to be for the psychopharmacological activity is essential, can be replaced by various other groups that the does not affect the biological activity of the peptides in question, but in some cases even to a considerable one Increase in activity.

It is the subject of the present invention, the amino acid residue L-Met with terminal N-atom of the key peptide 4-6 ACTH and the (on the side with terminal! G-atom) extended Modify peptides and peptide derivatives as mentioned above to the following groups:

D-methionyl, L- or D-methionyl sulfoxide (Met- »· O), L- or D-methionyl sulfone (Met- * Op), desaminomethionyl or the equivalent Sulfoxide or sulfone thereof or the group HpN-B-C-,

0 in which B is a branched or unbranched alkylene group having 1-6 carbon atoms. The last-mentioned group (HpN-B ₁ -C-) comprises amino acid residues as well as amino-substituted ones

.0

Carboxylic acid residues such as GIy ₁ VaI, AIa, β-Ala and (oC-Me) Ala.

• They are purely from a purely synthetic-chemical point of view Peptides in which L-Met has been replaced by one of the above groups which do not have a center of asymmetry, such as Desamino-methionyl, desamino-methionyl-sulfone, GIy or ß-Ala, preferred because these peptides can be more easily produced.

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Apart from these synthetic-chemical considerations, the modifications of L-Met lead to methionyl sulfoxide (L- or D-Met- »O), methionyl-sulfone (L- or D-Met-» O _? ), Desaminomethionyl, desaminomethionyl sulfoxide, Desaminomethionyl sulfone and β-Ala lead to a substantial increase in activity compared to the corresponding L-Met peptide, so that these modifications are particularly preferred. The invention therefore relates to new and valuable psychopharmacological peptides or peptide derivatives of the general formula

A-L-GIu-L-His-X I

in the A HD-Met, HL-Met (- »Ο), HD-Met (-K)), HL-Met (-» ög), HD-Met (- ^ Op), Desamino-Met, Desamino-Met (- »0), Desamino-Met (- * O ₂ ) or the group H-NH-B-CO-, in which B is a branched or unbranched alkylene group with 1-6 carbon atoms and X is a hydroxyl group, a (N- Phenylalkyl) amino group of the general formula _u

-NH - Alk -

in R. represents a hydrogen or halogen atom, a hydroxy group-alkyl group having 1-4 carbon atoms or alkoxy group having 1-4 carbon atoms, and Alk is a branched or unbranched alkylene group having 1-6 carbon atoms is _t or the group L-Phe-Y, in the Y is a hydroxyl group, an (N-aminoalkyl) amino group of the general formula -NH-AIk-NRgR ,, in the Alk

a branched or unbranched alkylene group with 2-6 carbon atoms, Rp a hydrogen atom or a lower alkyl group with 1-6 carbon atoms and R ^ a hydrogen atom or a lower alcyl group with 1-6 carbon atoms or an amidine group or the group L-Lys-Z or L-Arg-Z, in which Z is a hydroxyl group, denotes the group L-Trp-OH, L-Trp-Gly-OH or an (N-3-indolylalkyl) amino group, the alkyl group containing 1-6 carbon atoms, as well as the functional parts

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Derivatives of these peptides and peptide derivatives.

The peptides and peptide derivatives according to the invention are produced by a method as is commonly used in peptide chemistry. The processes as they are customarily used for the preparation of the compounds according to the invention can be summarized as follows;

1. Condensation of a compound (amino acid, peptide) with a free carboxyl group and protected other reactive ones Groups with a compound (amino acid, peptide or amine) with a free amino group and protected other reactive ones Groups in the presence of a condensing agent $

2. Condensation of a compound (amino acid, peptide) with an activated carboxyl group and possibly protected other reactive groups with a compound (amino acid, Peptide, amine) with a free NHp group and protected other reactive groups ^

3. Condensation of a compound (amino acid, peptide) with a free carboxyl group and protected others reactive groups with a compound (amino acid, peptide, amine) with an activated amino group and optionally protected other reactive groups,

whereupon the protection groups are removed if necessary.

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Activation of the carboxyl group can be achieved, for example, by converting the carboxyl group to an acid halide, acid _T anhydride or imidazolide group or an activated ester group such as the N-hydroxysuccinimide ester group and the p-nitrophenyl ester group.

The amino group can be activated by conversion to a phosphitamide or by the "phosphorus-azo" method.

The usual procedures for the condensation reactions given above are: the carbodiimide method, the azide method, the mixed anhydride method and the method the activated esters (E. Schröder and K. Lübke in "The Peptides", Vol. I, 1965 (Academic Press)). Merrifield's "Solid phase method" (J. Am. Chem. Soc. 8 £, 214-9 (1963)) for Production of the peptides and peptide derivatives according to the invention are used.

The reactive groups that are not involved in the condensation reaction should participate, are effectively protected by the so-called protective groups, which are easily removed again can be, e.g. by hydrolysis or reduction. So z. B. can be effectively protected by a carboxyl group Esterification with methanol, ethanol, tert-butanol, benzyl alcohol or p-nitrobenzyl alcohol or by conversion to a Amide. However, this last-named protective group can only be removed with great difficulty, so that it is advisable to use this Group only to be used to protect the carboxyl group of the C-terminated amino acid in the last peptide /. In In this case the peptide synthesis leads directly to the amide of the peptide of the general formula I.

Groups which are suitable for the effective protection of an amino group are usually acid groups e.g. an acid group, which is derived from an aliphatic, aromatic

/ or the ^ - carboxyl group of glutamic acid

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see, araliphatic or heterocyclic carboxylic acid, such as acetic acid, benzoic acid, fyridine carboxylic acid or an acid group which is derived from carbonic acid _/ such as an ethoxyoarbonyl, benzyloxycarbonyl, tert-butyloxycarbonyl or p-methyloxybenzyloxycarbonyl group or an acid group which is derived from a sulfonic acid such as the p-benzenesulfonyl or p-toluenesulfonyl group. However, other groups can also be used, such as substituted or unsubstituted aryl or aralkyl groups, for example benzyl and triphenylmethyl groups or groups such as the ortho-nitro-phenylsulfenyl or 2-benzoyl-1-methylvinyl group.

It is highly recommended to also visit the guanidino group of the Arginine, the £ -amino group of lysine and the imidazole group of histidine, although this protection is not absolutely necessary. Favorable protective groups in this context are a tert-butyloxycarbonyl or a toayl group for the £ -amino group of Lysina, a nitro group for the guanidine group des arginine and a benzyl, dinitrophenyl or a Trityl group for the imidazole group of histidine.

The protective groups can be split off by various customary methods, depending on the nature of the respective Group, e.g. with trifluoroacetic acid or by mild Reduction, e.g. with hydrogen and a catalyst such as Palladium or with HBr in glacial acetic acid.

Peptides according to the invention which, as a group with a terminal nitrogen, are a methionyl sulfoxide or desaminomethionyl sulfoxide group can be prepared from the corresponding Met or Desamino-Met peptide using an an known mild oxidation processes, e.g. with dilute hydrogen peroxide or a peracid. Such a one

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Oxidation leads to a mixture of the S and R sulfoxides (β 1- or d-sulfoxide), which is common in the individual Diaetereomers can be split.

When coupling S- or R-sulfoxide (1- or d-sulfoxide) of methionine or deaminomethionine with the peptide Glu-His-X, where X has the meaning given above, the individual enantiomers can also be obtained directly.

The peptides according to the invention, which as a residue with terminal nitrogen is a methionyl sulfone (Met * Op) or Desaminomethlonylsulfon (Desamino-Met- ^ Op) group, can be most conveniently prepared by a known per se oxidation of the corresponding Met- or desamino-Met-peptide, e.g. with HpOp or a peracid.

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. ₁₀ _ 23Q8362 _U _ _{42 571}

Among functional derivatives of the peptides according to the invention and peptide derivatives are to be understood:

1 · Pharmaceutically suitable acid addition salts of the peptides and peptide derivatives;

2. Peptides or peptide derivatives in which one or more free amino groups have been replaced by an acyl group, which is derived from an aliphatic carboxylic acid having 1-6 carbon atoms such as an acetyl group;

3. Unsubstituted amides or amides substituted by alkyl groups having 1-6 carbon atoms of the invention Peptides and peptide derivatives that have a free carboxyl group)

4 x esters of the peptides of the invention that are derived of aliphatic or araliphatic alcohols with 1-1 θ carbon atoms, such as methanol, ethanol, pentanol, Hexanol, cyclohexanol, octyl alcohol, undeoyl alcohol, hexadecyl alcohol, Oleyl alcohol, octadecyl alcohol, benzyl alcohol, phenylethyl alcohol, phenylpropyl alcohol or cinnamyl alcoholj

5 · Metal complexes that have been formed by bringing the peptides or peptide derivatives together with a little soluble salt, hydroxide or oxide of a metal, preferably zinc, preparations that have been manufactured by bringing the peptides according to the invention together with organic, mostly polymeric substances such as gelatin, Polyphloretin phosphate or polyglutamic acid to achieve a prolonged effectiveness.

The acid addition salts are obtained by reacting the peptides according to the invention with a pharmaceutically suitable organic or inorganic acid such as HCl, phosphoric acid, acetic acid, maleic acid, tartaric acid or citric acid.

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1A-42

As mentioned briefly above, the compounds according to the invention and their functional derivatives are valuable psychopharmacological activities. The inventive Compounds inhibit the extinction of the evasive reaction. That means they are general can be used as anti-depressant agents. In particular, they can be used to treat certain Mental disorders, where a stimulation of the mental reaction (mental performance) is desired, like in certain types of neurosis and in old age

(Senility). «Rands - _; stu3 -

t on the “uALi

The peptides according to the invention and the derivatives given above can be administered orally, parenterally or intranasally will. The peptides are preferably used as an injection preparation, for which they are in a suitable liquid be dissolved, suspended or emulsified. But they can also be mixed with suitable auxiliaries and fillers in a form suitable for oral administration such as pills, tablets or dragees. The peptides according to the invention can also be administered in the form of suppositories or sprays.

The peptides are preferably administered in daily doses of 0.001 to 1 mg / kg body weight, depending on the Activity and the form in which they are administered. Particularly favorable preparations are obtained when the peptides according to the invention are prepared in a form, in which they have a longer effectiveness, e.g.

O/
builds in gelatine, plypholoretin phosphate or preferably

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230§362 u-42 571

in the form of metal complexes. These metal complexes can be obtained by treating the peptides with poorly soluble Metal salts, metal hydroxides or metal oxides brings together. Are as poorly soluble metal salts commonly the metal phosphates, metal pyrophosphates and metal polyphosphates are used.

Metals that can be used for this process are the metals that belong to the b groups of the periodic table include, e.g., cobalt, nickel, copper, iron and preferably Zinc as well as the metals that belong to the main groups of the periodic table and can form complexes, such as magnesium and aluminum.

The production of these metal complexes takes place in the usual way. They can be obtained by adding of peptides and a sparingly soluble metal salt, metal hydroxide or metal oxide to an aqueous medium. They can also be obtained by adding an alkaline one Medium to an aqueous solution of the peptide and an insoluble metal salt, whereby an insoluble peptide-metal hydroxide complex arises. In addition, the metal complexes can be obtained by adding the peptide, one soluble metal salt and a soluble salt to an aqueous, preferably alkaline medium, wherein the insoluble peptide-metal salt complex in situ.

The metal complexes can be used immediately as suspensions or, for example, lyophilized and then resuspended will.

Biological effectiveness, /

^B. yiHitKt *

a) Elimination of the evasive reaction · Male white rats, weighing approximately 15o g, were made with the help of the so-called pole jumping test

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conditioned. The conditioned stimulus was a light above the cage, upon which the unconditioned stimulus of a shock is transmitted after 5 seconds the lattice floor of the cage followed. On three consecutive days, 10 tests per day were carried out with an average interval of 60 seconds. After this learning period, the trigger in studies of 10 individual experiments examined. All those animals that reacted 8 or more times in the first attempt at extinction, were treated with the substance under investigation or a placebo. Subsequently, extinction studies were carried out of 10 individual experiments each 2 and 4 hours after the treatment of the animals with the to be examined Substance carried out.

In the following table the results with the known peptide 4-10 ACTH are compared with some according to the invention Peptides.

- H-

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2308362
Peptide or
Peptide derivative dose
ug per
animal
S.C. 14 -
1. Under
search
Oh ?.Under-
3UChung
2 h 3. Under
search
4 h 1A-42 571
calculated
relationship
the effect
sameness
(4-10 ACTH
= 1) H-Met-Glu-Hii'-Phe-
Are-Trp-Gly-OH 100 ») 8th 7th 30th 9 7th 10 9 6th 3 r-I I H-D-Met-Glu-His-OH 30th 8th 6th 5 1 H-Val-Glu-His-OH 30th 8th 6th 1 H-ß-Ala-Glu-His-OH 30th 9 8th 7th 10 9 7th 6th 3 H-Met (- »O) -Glu-His-OH 30th 8th 8th 8th 10 8th 8th 7th 3 8th if 2 > 3 H-D-Met-Glu-His-
Phe-OH 30th 8th 7th 5 10 8th 6th 2 1 H-D-Het-Glu-His
Phe-Lys-OH 30th 8th 7th 5 1 H-D-Met-Glu-His-
Phe-Lys-Tra 30th 10 10 8th 10 8th 8th 7th 3 9 ■ 8th 7th > 10 H-Met (- »O) -Glu-His-
Phe-Arg-Trp-Gly-OH 10 8th 8th 8th 3 8th 2 > 3 H-Met (d, - »0) -Glu-His-
Phe-Arg-Trp-Gly-OH 5 9 7th 6th 1.5 9 7th .3 6th H-p-Ala-Glu-Hia-Phe-
Arg-Trp-Gly-OH 30th O 8th 8th ;O 9 8th b > 3 H-D-Met-Glu-His-Amf , 30 8th 8th 6th 10 9 7th 3 > 1 Deaamino-Met-Glu-His-
Phe-Lys-Trp-OH 30th 9 8th 8th 10 9 7th 6th 3 9 5 3 5 Deeamino-Met (- »0-) -
Qlu-His-Phe-Lys-Trp-
OH 10 8th 8th 8th 3 9 6th 5 1 8th 5 2. 10

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The following applies to the description, the examples and the claims:

I. If no optical configuration is mentioned, the L-shape is meant.

II. The following abbreviations are used for the protecting groups or activating groups.

Z = Benzyloxycarbonyl Boo = tert-butyloxycarbonyl tBu = tert-butyl Me = Methyl ONP = p-nitrophenyloxy ONB = p-nitrobenzyloxy Su = Succinimido

III. The following abbreviations are used for solvents and reagents.

Bz = benzene BtOH Ethanol Bu Butanol Py Pyridine Ao or HAc = acetic acid Wa water On = Amyl alcohol iPro = Isopropanol DMF « Dimethylformamide THF Tetrahydrofuran DCCI Dicyclohexylcarbodiimide DCHU Dicyclohexyl urine ο ff TAA ■ Triethylamine TFA Trifluoroacetic acid

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1A-42

IV. FUET amino acid residues The following abbreviations

applied. Methionyl Mead Methionyl sulfoxide (Raoemat) Mead (- »Ο) Methionyl (d) sulfoxide Met (d, -> 0) Methionyl (1) sulfoxide Met (1, * O) Methiοny1sulfοη Met (* O ₂ ) Glutamyl GIu Glutaminyl Gin or GIu (NH ₂ ) = Histidyl Hia Phenylalanyl Phe Arginyl Arg = Lysyl Lys = Tryptophyl Trp Glyoyl GIy Valyl VaI AlanyI AIa β-alanyl ß-Ala «C-methylalanyl (oC-Me) Ala

Abbreviations for other residues Tra =

Desamino mead

Amf

(N-3-indolylethyl) amino group (derived from tryptamine)

Desamino-methionyl (or Γ-methylthiobutyryl) group

(N-l-phenylisopropyl) amino group (derived from amfetamine).

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example 1

A. Η-Met (- »Ο) -GIu-Hi a-OH

25 mg of the tripeptide H-Met-Glu-His-OH were dissolved in 2.5 cm ^{3 of} acetic acid and then 15% of hydrogen peroxide was added. After stirring at 2O ⁰ G a suspension of 20 mg of platinum sohwarz added in 2.5 cm of glacial acetic acid and the mixture stirred 30 min. The mixture was then filtered and the solvent was distilled off in vacuo. The resulting residue was taken up in 10 cm tert-butanol / water (1: 1) and lyophilized.

Rf of the sulfoxide = 0.14 in Bu: Py: Ac: Wa (4: 3/4: 1/4: 1) on SiO ₃ .

Example 2

Synthesis of D-Met-Glu-His derivatives and desamino-Met-GIu-His derivatives

A. Boo-D-Met-Glu (OtBu) -HiS-OMe

10.52 g Boc-D-Met-N ^ H, in 75 cm ⁵ DMP ⁰ C were cooled to O and then 23.6 cm 3.4 η hydrochloric acid in THF was added and stored at -20 ⁰ C 5.85 cm Isoamyl nitrite. The mixture was stirred for 7 min and then 17.05 g of H-GIu (OtBu) -Hie-OMe.2 HCl in 50 cm 'DMP were added and the pH was adjusted to 6.9 with triethylamine. The mixture was stirred for a further 3 days at 0 ° and then the mixture was filtered and the piltrate was evaporated to dryness in vacuo. The residue was taken up in 150 cnr ethyl acetate / water and washed with water

• 5 washed. The organic phase was dried, evaporated to 100 cm and set aside at ⁰ ° C. Hf in Bu: Ao: Wa (4: 1: 1) = 0.63 (SiO ₂ ). Mp 69-71 ⁰ C.

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B. Desamino-Met-Glu (OtBu) -Hi s-OMe I.

In the same way as described under 2 A. the 'Desamino-Met-NpH, converted into the azide and then coupled with H-GIu (OtBu) -HiS-OMe.2 HCl, after which the compound obtained was isolated and purified, as stated under A. Rf in Bu: Ac: Wa (4: 1: 1) = 0.50

C. Boo-D-Met-GIu (OtBu) -HiS-OH

1.17 g tripeptide ester (Example 2 A.) was placed in 50 cm Dissolved 50 # dioxane. 2.5 cm ^ 1 η Added sodium hydroxide solution and then the mixture was stirred at 20 G for 30 min. The reaction mixture was on a pH value of 7 neutralized and then evaporated to dryness in vacuo. The residue was dissolved in 40 cm of ethyl acetate added, acidified to a pH of 4 with 1.4 cm 2 η HGl and washed with a little water and then separated the layers.

The Ethylaoetatsohioht was dried and evaporated in vacuo to Trookene. The residue was an oil. Rf in Am: Py: Wa (5: 3: 2) = 0.19 (SiO ₂ ).

D. Boc-D-Met-Glu (HH ₂ ) -His-0H

1 g of tripeptide ester was dissolved in 20 cm of dioxane / water (1: 1) with heating. The mixture was cooled to 20 ⁰ C and then 1.9 meq ^ barium hydroxide was added and stirred the Gemisoh 30 min. After neutralizing to a pH of 5 | 4, the Gemisoh was carefully evaporated. The residue was dissolved in methanol, filtered and the filtrate was diluted with ethyl acetate.

The precipitate was filtered off and recrystallized from methanol / ethyl acetate.

Rf in Am: iPro: Wa (10: 4: 5) = 0.59 (SiO ₂ ) ...

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E. Boo-D-Met-Glu (OtBu) -Hi3-NH ₂

1 g of Boo-D-Met-Glu (0tBu) -Hi3-0Me (Example 2A) was dissolved in a cooled solution of dry ammonia gas in 25 cm of methanol. The mixture was stirred for 25 h and then the ammonia was evaporated, the residue was taken up in methanol and the solution was diluted with ether. The precipitate formed was filtered off at.

Mp 72-75 ⁰ C. Rf in Bz: BtOH (8: 2) = 0.43 (SiO _2).

F. Desamino-Met-Glu (0t Bu) -HIs-OH

The ester obtained in Example 2 B. was hydrolyzed in the manner described in Example 2 C.

Rf in Am: Py: Wa (5: 3: 2) = 0.16 (SiO ₂ ).

G. Removal of the protecting group (s) from the feptides after AF

100 mg of the peptides (A, B, C, D, E or F) were in 5 cnr 90? Bigem TFA solved. The solution was left to stand for 1 hour and then the solvent evaporated, the residue stirred twice with ether and over solid potassium hydroxide dried. The trifluoroacetate was dissolved in 20 cm tert-butanol / Water (1: 1) and then dissolved by stirring the solution with so much Dowex X-8 in the acetate form that you get a final pH of 5 | 4 was converted into the acetate. After filtering the filtrate was lyophilized and the white residue was stored over solid phosphorus pentaxide.

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received product

t 2308362
1A-42 571 Starting
peptide Rf in
Bu: Py: Ao: V / a (4: 3/4: i / 4: 1)
on SiO ₂ 2 A. 0.18 2 G. 0.14 2 B. 0.25 2 p. 0.21 2 D. 0.17 2 E. 0.16

1. H-D-Met-Glu-His-OMe.HAo

2. H-D-Met-Glu-His-OH.HAc 3 • Desamino-Met-Glu-His-OMe

4. Desamino-Met-Glu-His-OH

5. H-D-Met-Gln-His-OH.HAc

6. HD-Met-Glu-His-NH ₂ .HAo

H. Sulphoxide of HD-Met-Glu-His-OH

25 mg of the tripeptide H-D-Met-GIu-His-OH (G. 2) were in 2.5 cm of acetic acid dissolved and then 15yul 30 $ iges Hydrogen peroxide added. The mixture was treated in the manner described in Example 1. The deficit was taken up in 10 cm * tert-butanol / water (1: 1) and then the solution is lyophilized.

Rf in Bu: Py: Ac: Wa (4: 3/4: 1/4: 1) = 0.12 on SiO ₂ .

K. Desamino-Met-Glu-His-OH sulfoxide

25 mg of the peptides obtained according to G. 4 were oxidized in the manner described under H.

Rf in Bu: Py: Ac: Wa (4: 3/4: 1/4: 1) = 0.18

Example 3

Synthesis of A ^ GIu-His derivatives (A = VaI, GIy, Ala, ß-Ala or

A. Boo-Val-Glu (OtBu) -HiS-OMe

3.26 @ (15 mMo]) Boc-Val-OH were dissolved in 20 cm ^{5 of} methylene chloride and then 1.73 g of N-hydroxysuccinimide were added. The solution was cooled to -2O ⁰ C and were subsequently

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3.09 g DCCI in 20 cm * I. chilled methylene chloride was added 'and the resulting solution for 1 h at -2O ⁰ C and then stirred for 20 h at + 2O ⁰ C.

The resulting DCHU was filtered, then the filtrate evaporated to dryness and the residue dissolved in 30 cm DMP, whereupon 7.33 g Z-GIu (OtBu) -HiS-OMe (Kappler HeIv. 44, 1991, 1961) and 1.4 g of 10 $ palladium on carbon were added. Hydrogen was then passed through for 5 hours and then the mixture was stirred and filtered for 1 hour and the piltrate was added Evaporated dry. The residue was dissolved in aqueous ethyl acetate dissolved and made with citric acid, water, sodium bicarbonate and washed water. The organic phase was dried and then the. Ethyl acetate evaporated in vacuo. The residue was recrystallized from ethyl acetate / petroleum ether.

Yield 3.95g; Pp 117-119 ⁰ C; / oil I ^ = -8.4 (O = I ₁ DMF); Rf in Bz: EtOH (8: 2) = 0.55 (SiO ₂ ).

B. Boo-Gly-Glu (OtBu) -His-OMe

Starting from 2.63 g of Boc-Gly-OH, the tripeptide Boo-Gly-Glu (OtBu) -His-OMe was prepared by the method described in Example A. Yield: 1.47 g; Pp 103-108 ⁰ C;

Rf in Bz: EtOH (8: 2) = 0.43 (SiO ₂ ).

C. BoO-AIa-GIu (OtBu) -HiS-OMe

3.78 g of Boc-Ala-OH were converted into the "active ester ¹¹ " with 2.3 g of N-hydroxysuccinimide and 4.12 g of DCCI. The active ester, dissolved in 40 cm ^{5 of} DMP, was condensed with H-GIu (OtBu ) - His-OMe, prepared from 9.77 g of Z-GIu (OtBu) -HiS-OMe, as described in Example 3 A.
Yield: 5.56 g; M.p. 97.5-101 ° C.
Hf in Bz: EtOH (8: 2) = 0.36 (SiO ₂ ).

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D. Boo-ß-Ala-Gl u ( OtBu) -HIs-OMe

Starting from 3.78 g of Boc-beta-Ala-OH (m.p. 77-78 ⁰ C) was Peptidester Boo-ß-Ala-Glu (OtBu) -His-OMe, as described in Example 3 C. manufactured.

Yield: 7.4 g; ^ fLj = -8.7 (c = 2, DMF). Rf in Bz: EtOH (8: 2) = 0.25 (SiQ ₂ ).

E. BoC-K-Me) AIa-GIu (OtBu) -HiS-OMe

1.83 g of Boc- (1-methyl) alanine (melting point 118-119.5 ° C.) were dissolved in 40 cm of methylene chloride and then 3.51 g of H-GIu (OtBu) -HiS-OMe.2 HCl were prepared by hydrogenating the Z-peptide in methanol, in the presence of 2 eq hydrochloric acid and 2.29 cm TAA was added to the solution. The mixture was cooled to 0 ° and then 2.07 g of N-Hydroxysuocinimid was added and finally, after the Gemisoh had been cooled to -20 ⁰ C, 1.86 g of DCCI. The reaction mixture was further treated in the above manner besohriebene (example A.) after at -2O ⁰ C ₁ 2 at ⁰ ° C and stirred for 20 hours at 2O ⁰ C was 10 min h.
Yield: 3.02 g of oil.
Rf in Bu: Ao: Wa (4: 1: 1) = 0.61 (SiO ₂ ).

F. Hydrolysis of peptide esters A, B, C, D and E

1.15 g of the peptide esters A, B, C or D were dissolved in 50 cm 50 tiger dioxane. After addition of 2.5 cnr 1 η Natriumhydroiidlöeung the mixture was stirred for 30 min at 2O ⁰ C. The reaction mixture was neutralized to a pH of 7 and then evaporated to dryness, the residue was taken up in 40 cm of ethyl acetate and the solution was washed with 0.7 cm 2 of hydrochloric acid (pH 4). The ethyl acetate was dried and evaporated to dryness.

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Rf in Am: Py: Wa (5: 3: 2) (SiO ₂ )

1. BoC-VaI-GIu (OtBu) -HiS-OH 0.24

2. Boo-Gly-Glu (0tBu) -His-0H 0.23

3. Boo-Ala-Glu (0tBu) -His-0H 0.23

4. Boo-β-Ala-Glu (0tBu) -His-0H 0.26

5. BoO- (OC-Me) AIa-GIu (OtBu) -HiS-OH 0.25

G. Removal of the protecting group !! of the peptides obtained according to F. 1 to F. 5.

In the manner described in Example 2 G., the in 5 F. described peptides with TFA freed from the protective group and converted into the acetate.

Acetate of: Rf in Bu: Py: Ac: Wa (4: 3/4: 1/4: 1)

on SiO ₂

1. H-VaI-GIu-His-OH 0.15

2. H-Gly-Glu-His-OH 0.15

3. H-Ala-Glu-His-OH 0.17

4. H-β-Ala-Glu-His-OH 0.17

5. H- (oC-Me) Ala-Glu-His-OH 0.16

H. Removal of the protective groups from the peptide esters according to A. to JE.

In the manner described in Example 2 G., the _{tripeptide esters obtained according to A 1} B, C, D and E were freed from the protective groups and converted into the acetate.

Acetate of Rf in Bu: Py: Ac: Wa (4: 3/4: 1/4: 1)

on

1. H-Val-Glu-His-OMe 0.20

2. H-Gly-Glu-His-OMe 0.18

3. H-Ala-Glu-His-OMe 0.21

4. H-β-Ala-Glu-His-OMe 0.22

5. H - («C-Me) Ala-Glu-His-OMe 0.18

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Example 4

Synthesis of HU-Me) Ala-Glu-Hls-R (R = -NH ₂ , -N (CHJ ₂ or -NHC ₂ H ₅ )

A. 0.7 g of Boc- (i * -Me) Ala-GIu (OtBu) -His-OMe (Example 3 E.) were dissolved in a cooled solution of dry ammonia gas in 20 cm of methanol. The mixture was stirred for 20 h and then the ammonia was evaporated, the residue was taken up again in methanol and the solution was diluted with ether. The resulting precipitate was filtered off (0.41 g) mp 93-95 ° C. The precipitate was then deduced in 20 cm. TPA solved. The solution was left to stand for 1 h, then the solvent was evaporated off in vacuo, the residue obtained was stirred in ether and dried over KOH pellets. The trifluoroacetate was then converted to the acetate by adding sufficient Dowex X-8 in the acetate form to a solution of the peptide in t-butanol / v / water (1: 1) to a pH of about 5.4. When the mixture was filtered and the piltrate was lyophilized, an almost white powder of pure peptide amide H-fct-Me) Ala-Glu-His-NH _? . Rf in Bu: Py: Ac: V / a (4i3 / 4: 1/4: 1) = 0.20 on SiO ₂ .

B. If instead of dry ammonia gas dimethylamine or Ethylamine was used, the corresponding N, N-dimethylpeptide amide was obtained or N-ethyl peptide amide.

Example 5 Synthesis of D-Met-Glu-His-Phe derivatives

A. Z-Glu (OtBu) -His-Phe-OMe

9.76 g of Z-GIu (OtBu) -HiS-N ₂ H were dissolved in 30 cm ^{5 of} DMP and the solution was then cooled to OC.

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Haoh the addition of 14.4 cm ⁵ 4.8 η hydrochloric acid / THF was further cool the solution to -2O ⁰ C and then 3.24 cm isoamyl nitrite were added dropwise. The solution was stirred for 7 min and then a solution of 4.32 g of H-Phe-OMe.HCl in 40 cm ^{5 of} DMP and 12.6 cm ^{3 of} TAA, which had been ^{precooled to 0} ° C., was added. The mixture was left to stand at ⁰ ° C. for 3 days and then filtered. This was taken up in ethyl acetate / water. The organic phase was washed successively with water, sodium bicarbonate solution and water and then dried and the solvent was distilled off in vacuo. The residue was recrystallized from hot ethyl acetate.

Mp 183 °, Z ^ 7.beta. ⁵ = -21 ° (° = 1 DMF). Hf in Bz: EtOH (8: 2) (SiO ₂ ) = 0.81.

B. Z-Glu (OtBu) -HiS-PhO-OH

^{3.3 g of phenylalanine and 12.6 cm of triethylamine in 15 cm of DMP at 0 °} C. were added to an equal amount of the azld Z-GIu (OtBu) -HiS-N ,, which had been prepared according to A. The mixture was ^{left to stand for 3 days at 0} ° C. and then filtered. The filtrate was evaporated to dryness, taken up in ethyl acetate / water, acidified to pH 3 with citric acid and then washed with water. The layers were separated and the organic layer dried and evaporated. *

The residue was recrystallized from aoetonitrile / ether. Rf in Am: Py: Wa (5: 3: 2) = 0.73 (SiO ₂ ).

C. Z-Glu (OtBu) -Hi s- (N-phenylethyl) amide

The phenylalanine used under B. was replaced by ß-phenylethylamine, resulting in the following peptide: Z-GIu (OtBu) -HiS- (N-phenylethyl) amide.

Rf in Bz: BtOH (9: 1) = 0.47 (SiO ₉ ).

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D. Z-Glu (OtBu) -His-Phe-NH

1 g of the Tripeptldestera described under A. was dissolved in 20 cm of methanol and then atmospheric gas was passed through the Gemisoh. The mixture was then stirred for 1 night and then the precipitate was filtered off. After recrystallization from the methanol / ether, the amide was obtained. Pp 119-120 ⁰ C
Hf in Bu: Py: Ac: Wa (4: 3/4: 1/4: 1) = 0.70 (SiO ₂ ).

E. Removal of the protective groups from the peptides or peptide derivatives naoh AD

0.7 g of the peptide obtained according to A, B, C or D were dissolved in 18 cm of pure methanol and then 10 $ palladium on carbon was added. Hydrogen was durohgeleitet by the Gemisoh until no more CO ₂ escaped and then the Gemisoh filtered and evaporated the PiItrat in vacuo at a bath temperature of 3O ⁰ C. The peptide, which had been partially deprotected, was processed further immediately.

Peptide: Rf in Am: Py: Wa (5: 3: 2)

(SiO ₂ )

1. H-Glu (OtBu) -His-Phe-OMe 0.35

2. H-Glu (OtBu) -His-Phe-OH 0.23

3. H-Glu (0tBu) -His-Phe-NH ₂ 0.47

4. H-Glu (OtBu) -His- (N-phenylethyl) amide 0.41

p. Condensation of D-Met with the peptides prepared in accordance with E.

3.91 g of Boo-D-Met-Ν ,, Η, were dissolved in 75 cm ^{3 of} DMP. The solution was cooled to OC and then 7.5 cm

4 η HOl / THP was added and the solution was further cooled to -20 ⁰ C. Then 2.1 cm of isoamyl nitrite were added and the Gemieoh

Stirred for 5 min and the volume made up to 90 om ("Solution A").

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In the meantime, 5 mmol of one of the tripeptides prepared in accordance with E. was dissolved in 20 cm of DMP and 2 cm of water. This solution was cooled to ^{0 ° C.}

30 om of the solution A was added to the tripeptide solution. The mixture was ^{left to stand at 0} ° C. for 72 h and then the DMP was distilled off and the residue was taken up in ethyl acetate / water. The organic phase was washed with bicarbonate solution and water and then dried and the solvent was distilled off in vacuo. The residue was recrystallized from methanol / ether.

Peptide: Rf in Bz: EtOH (8: 2) on SiO ₂

1. Boc-D-Met-Glu (OtBu) -His-Phe-OMe 0.42

2. Boc-D-Met-Glu (OtBu) -His-Phe-OH 0.14

3. Boo-D-4iet-Glu (0tBu) -His-Phe-NH ₂ 0.20 4 * Boc-D-Met-Glu (OtBu) -His- (N-phenylethyl) amide 0.40

G · Condensation of desamino-Met with the peptides prepared in accordance with E.

In the manner described under F. 5 mmol of deaminomethionyl hydrazide with the help of isoamyl nitrite into the corresponding Converted azide and then coupled the azide formed with one of the peptides produced under E.

Peptide: Rf in Bz: EtOH (8: 2) on SiO ₂

1. Desamino-Met-GIu (OtBu) -His-Phe-OMe 0.36

2. Desamino-Met-Glu (OtBu) -His-Phe-OH 0.11

3. Deaamino-Met-Glu (0tBu) -His-Phe-NH ₂ 0.17

4. De8amino-Met-Glu (OtBu) -His- (N-phenylethyl) -

amide 0.34

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H. Boc-D-Met-GIu (Ot Bu) -HiS-PhB-OC ... H ^

I I-O

0.25 g of the peptide Boc-Met-Glu (OtBu) -His-Phe-OH (prepared according to P. 2) were in 1.4 cm Undeoanol and 0.3 g Dowex X-8.HC1 (starting resin) solved. The mixture was stirred and filtered for 7 days at room temperature and the filtrate was evaporated in vacuo. The undecyl ester was isolated by chromatography of the residue obtained over SiOp with the aid of benzoil / ethanol (9: 1) as the eluent.
Rf in Bz: EtOH (8: 2) = 0.68 (SiO ₂ ).

K. Removal of the protective groups of the peptides obtained under F, G and H.

In a manner similar to that described in Example 2 G., the peptides obtained under F, G and H were removed from the protective groups freed.

Peptide: Rf in Bu: Py: Ac: Wa (4: 3/4: 1/4: 1)

on

1. H-D-Met-GIu-His-Phe-OH 0.27

2. H-D-Met-Glu-His-Phe-OMe 0.35

3. H-D-Met-GIu-Hi8- (N-phenylethyl) amide 0.36

4. HD-Met-GIu-His-Phe-NH ₂ 0.33

5. H-D-Met-Glu-His-Phe-OC ^ H ^ 0.39

6. Desamino-Met-Glu-His-Phe-OMe 0.40

7. Desamino-Met-Glu-His-Phe-OH 0.31

8. Desamino-Met-Glu-His-amide 0.38

9. Desamino-Met-Glu-His- (ii-phenylethyl) amide 0.42

L. Sulfoxide

30 mg of the tetrapeptide H-Met-Glu-His-Phe-OH were dissolved in 2.5 cm ^{5 of} acetic acid and then 15 μl of 30% HpOp were added. The mixture was based on that in Example 1

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described way further processed. The residue was taken up in 5 om tert-butanol / water (1: 1) and the mixture was then lyophilized.

The peptides obtained under K. 1, L 2 and K. were converted into the corresponding sulfoxide in the same way.

Peptide; Rf in Bu: Py: Ac: Wa (4: 3/4: 1/4: 1)

on

H- * et (- »O) -Glu-His-Phe-0H 0.26

HD-Met 0 * O) -Glu-His-Phe-OH 0.26

HD-Met (- »0) -GIu-Hi 3-Phe-0Me 0.33

Desamlno-Met (- * 0) -Glu-His-Phe-OH 0.29

Example 6

Synthesis of Val-Glu-His ~ Phe derivatives

A. Boo-Val-Glu (OtBu) -Hls-Phe-NH ^

0.33 g of Boc-Val-OH were dissolved in 20 cm DMF and then 0.81 g of H-Glu (OtBu) -His-Phe-NH ₂ (Example 5 E. 3) were added and the pH was adjusted to 7 _f 2 set. After addition of 0.31 g of DCCI, the mixture was stirred for 5 h at ⁰ G and 20 h at 20 ⁰ C. The resulting DCHU was filtered off and then

3 sodium
the filtrate was poured into 100 cm 0.1N carbonate solution and the precipitate was stirred at ⁰ ° C. for 1 h. After filtering off, the chopping stand was dried and recrystallized twice from ethyl acetate / ether.

Yield: 0.51 g, mp 137 ° (dec.)

Hf in Bz: EtOH (6: 2) = 0.27 on SiO ₂ .

B. H-VaI-GIu-HIs-PhB-NH ₂

The removal of the protective group from the protected tetrapeptide amide according to A. by the method described in Example 2 G. resulted in the acetate of the peptides indicated above. _ 30 -

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Example 7 Synthesis of D-Met-GIu-His- (N-phenylalkyl) amides

1.17 g Boo-D-Met-Glu (OtBu) -His-N ₉ H, (Example 9 A.)

3
were dissolved in 20 cm DMF. This was cooled and then Lö3ung om 3 ⁵ 2 η HCl / THF at ⁰ ° C and 0.27 cm ⁵ isoamyl nitrite at -2O ⁰ G was added and the mixture stirred for 7 min at -2O ⁰ G.

3 mmol of the corresponding amine were added to the azide solution added and the pH adjusted to 7 with TAA.

The reaction mixture was ^{stirred at 0} ° C. for 70 h and then evaporated to dryness in vacuo and the residue was dissolved in aqueous ethyl acetate.

The organic phase was washed with water, bicarbonate solution and Wasθer and dried.

The residue was evaporated to dryness, then stirred with ether / petroleum ether and dried again.

The protective groups were removed according to the procedure described in Example 2 G.

Amine H-D-Met-Glu-His- (N-phenylalkyl) -

amid.HAc

Rf in Bu: Ao: Wa (4: 1: 1)

Benzylamine 0.20

Phenylethylamine ** 0.21

1-amphetamine 0.19

p-hydroxyphenylethylamine 0.19

p-methylbenzylamine 0.20

*) see Example 5 K. 3

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Example 8

Synthesis of A-Glu-His- (N-phenylalkyl) amides (A = VaI or ß-Ala)

A. 1.11 g BoC-VaI-GIu (OtBu) -HiS-OMe (Example 3 A.) were in

3 ^

Dissolved 20 om methanol and then added 1 cm hydrazine hydrate . The mixture was stirred at room temperature for 6 h, then the methanol was distilled off in vacuo and the residue that remained was stirred with water and dried.

The trookene residue was dissolved in DMF and coupled with 1-amphetamine (1- phenylisopropylamine) according to the azide method described in Example 7 enclosed.

In the manner described in Example 2 G., the resulting compound was freed from the protective groups, giving the peptide H-VaI-Glu-His- (Nl-phenylisopropyl) amide acetate, which was lyophilized.

Rf in Bu: Ao: Wa (4: 1: 1) = 0.18 on SiO ₂ * starting substance 0.53

B. Boo-ß ~ Ala-Glu (OtBu) -HiS-OMe was converted to the corresponding H-ß-Ala-Glu-His- (N-benzyl) amide in the manner described under A., instead of 1-amphetamine now Benzylamiη was used.

Rf in Bu: Ac: Wa (4: 1: 1) = 0.16 on SiO ₃ .

Example 9 Synthesis of D-Met-GIu-His-Phe-Arg

A. Boo-D-Met-Glu (OtBu) -HJs-N ₂ H,

3.2 g of Boc-D-Met-Glu (OtBu) -His-OMe (Example 2A) were Dissolved in 70 oar of methanol and then 3.7 cm of hydrazine hydrate were added. The mixture was left at room temperature for 5 hours stirred, then the methanol was distilled off in vacuo and the residue was stirred with water.

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After drying, the hydrazide was processed further immediately.

Rf in Am: iPro: Wa (10: 4: 5) = 0.37 on SiO ₂ .

B. Boo-D-Met-Glu (OtBu) -His-Phe-Arg-OH

1.17 g of the tripeptide hydrazide (A.) were dissolved in 20 cm ^{5 of} DMF. After addition of 3 cm ⁵ 2 η HCl in THF at ⁰ ° C the mixture was cooled further to -2O ⁰ C and then 0.27 cm Ieoamylnitrit added. The Gemisoh was stirred for 6 min and then the azide solution was gelatinized to 2 mmol of the peptide H-Phe-Arg-OH in 5 cm ⁵ DMF
was discontinued.

given in 5 cm DMF, whereupon the pH value with TAA to 7.3

The reaction mixture was ^{stirred at 0} ° C. for 70 h and then evaporated. The residue was stirred in ethyl acetate. An amorphous pentapeptide was obtained on evaporation of the ethyl acetate.

Rf in Bz: EtOH (8: 2) = 0.17 on SiO ₂ -

C. HD-Met-Glu-His-Phe-Arg-OH

Removal of the protective groups of the pentapeptide obtained under B. with trifluoroacetic acid gave the pentapeptide diacetate after exchange with Dowex X-8 in the acetate form.

Rf in Bu: Py: Ac: Wa (4: 3/4: 1/4: 1) = 0.18

Example 10

Sulfoxide of H-Met-Glu-His-Phe-Arg-OH, H-Met-Glu-His-Phe-Lys-OH and H-D-Met-Glu-His-Phe-Arg-OH

In the manner described in Example 1, the pentapeptides H-Met-GIu-His-Phe-Arg-OH, H-Met-Glu-His-Phe-Lys-OH and HD-Met-Glu-His-Phe-Arg- OH (Example 9 C.) oxidized in acetic acid with the aid of 30 $ H ₉ O _9.

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Peptide Hf in Bu: Py: Ao: Wa (4: 3/4: 1/4: 1)

on

Sulphoxide of H-Met-Glu-Hia-Phe-Arg-OH 0.18 Sulphoxide of H-Met-Glu-Hia-Phe-Lys-OH 0.21 Sulphoxide of H-D-Met-Glu-Hia-Phe-Arg-OH 0.17

Example 11

Synthesis of A-Glu-Hia-Phe-Lys derivatives (A = <X.-Me-Ala, AIa

fl-ala or desamino-mead)

A. H-Phe-Lys (Boo) -OtBu

For coupling of 4.2 g Z-Phe-ONP with H-Lys (Boc) -OtBu to give the dipeptide Z-Phe-Lys (Boo) -OtBu as a viscous oil.

Rf in Bz: EtOH (9: 1) = 0.57 on SiO ₂ ; Yield 79%.

The dipeptide H-Phe-Ly8 (Boo) -Ot3u was in 95 # yield obtained by hydrogenation of the peptide Z-Phe-Lya (Boc) -OtBu in methanol with 10 # palladium on carbon.

Rf in Bu: Ao: Wa (4: 1: 1) = 0.41

B. Boo-te-Me) AIa-GIu (OtBu) -HiB-N ₂ H ₃

2.9 g of the protected peptide Boc (f-Me) -Ala-Glu (OtBu) -His-OMe (Example 3 E.) were converted into the hydrazide with hydrazine hydrate in the manner described in Example 9 A. Fp 21Θ - 225 ⁰ C (dec.).

C. In the manner described under B, the methyl esters Boc-Ala-Glu (OtBu) -HiS-OMe (Example 3C), Boc-β-Ala-Glu (OtBu) -His-OMe (Example 3 D.) and desamino-Met-Glu (OtBu) -HiS-OMe (Example 2 B.) converted into the corresponding hydrazides.

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30983G / 1 185

D. HK-Me) AIa-GIu-HIa-PhB-LVa-OH

1 mmol of peptide hydrazide corresponding to B. was dissolved in 10 cm DMF. The mixture was cooled, then 3 cm 1 of HCl in THF and 0.13 cm of Iaoamyl nitrite were added. The mixture was then stirred ^{at -20 0 G for 6 min.} The azide obtained in this way was added to a solution of 1 mmol of H-Phe-Lya (Boo) -OtBu (from A.) in 5 om ⁵ DMF. After the pH had been adjusted to 7.4 with TAA, the mixture was ^{stirred at 0} ° C. for 70 h. The DMF was distilled off in vacuo, the Rüokatand dissolved in aqueous ethyl acetate and the organic phase washed with water, bicarbonate solution and water. The ethyl acetate was dried and distilled off in vacuo. The residue was chromatographed on SiOp and the peptide Boo- (oO-Me) Ala-Glu (OtBu) -His-Lys (Boo) -0tBu was isolated.

Hf in Bz: EtOH (8: 2) = 0.74 on SiO ^.

Removal of the protective groups from the protected peptide in the manner described in Example 2 G. gave the above pentapeptide acetate in 63% yield.

Rf in Bu: Py: Ao: Wa (4: 3/4: 1/4: 1) = 0.18 on SiO,

'2 *

£. In the manner described under D., the peptide methyl esters, which had been obtained according to C., converted into the corresponding azide and coupled with H-Phe-Lys (Boc) -OtBu (A).

When the protective groups were removed in the manner described in Example 2 G., the following non-protected ones were obtained Peptides as acetates in 60-70 # yield.

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Peptide: Rf in Bu: Py: Ao: Wa (4: 3/4: 1/4: 1)

on SiO ₂

H-β-Ala-Glu-His-Phe-Lys-OH 0.18

H-Ala-Glu-His-Phe-Lys-OH 0.20

Desamino-Met-Glu-His-Phe-Lys-OH O.24

H-D-Met-Glu-His-Phe-Lys-OH 0.18

Example 12

Synthesis of A-Glu-Hie-Phe-NH-CCH g ^ -NHp (A = D-Met, GIy or Desamino-Met)

Α- HD-Met-Glu-Hls ^ Phe-NH- (CH ₂ ) ,, - NH ₂

1.17 g of the tripeptide hydrazide Boc-D-Met-Glu (OtBu) -HiS-N ₂ H ^ 9A) were dissolved in 20 cm of DMF. After addition of 3 cnr 2 η HCl in THP at 0 ⁰ C, the Gemisoh was further cooled to -2O ⁰ C. Then 0.27 µm isoamyl nitrite was added to this mixture and the mixture was then stirred for some time. The azide solution obtained was then added to 2 mmol of H-Phe ~ NH- (CH ₉ ) _t --N-Boc

(obtained by hydrogenation of the corresponding compound Z-Phe-NH- (CH _{2) 5} -NH-Boc, m.p. 131-133 ⁰ C) cm ⁵ of DMF was added in 5 and then adjusted the pH with TAA at 7 _f. 5

The reaction mixture was ^{stirred at 0} ° C. for 70 h and then evaporated, and the residue was stirred in ethyl acetate. After evaporation of the ethyl acetate, a yellowish-white amorphous powder was obtained, which was treated further in the manner described in Example 2 G. Treatment with Dowex X-8 in the acetate form gave the HD-Met-Glu-His-Phe-NH- (CH ₂ ) ₅ -NH ₂ -diacetate. Rf in Bu: Py: Ac: Wa (2: 3/4: 1/4: 1) = 0.35 on SiO ₂ -

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- 36 - 1A-42 571

B. H-Gly-Glu-His-Phe-NH- (CH ₂ ) - NH ₂

Boc-Gly-Glu (OtBu) · HIe-NpH, (obtained from the corresponding methyl ester) was converted into the diacetate of H-Gly-Glu-His-Phe-NH- (CH ₂ J ₅ -Rf in Bu: Py: Ac: Wa (2: 3/4: 1/4: 1) = 0.32

C. Deeamino-Met-Glu-His-Phe-NH- (CH ₂ ) r -NH ₂

Desamino-Met-GIu (OtBu) -HiS-N ₂ H ₅ (Example 9 C.) was converted into the acetate of desamino-Met-GIu-His-Phe-NH- (CH ₂ ) C-NH ₂ . Rf in Bu: Py: Ac: Wa (2: 3/4: 1/4: 1) = 0.38.

Example 13 Synthesis of Val-Glu-His-Phe-Arg-Tra

A. Boc-Val-GIu (OtBu) -HJs-N ₂ H,

Starting from the tripeptide ester (Example 3 A.), the hydrazide BoC-VaI-GIu (OtBu) -HiS-N ₂ H _{3 was} based on the θ Α in example. way described.
Rf in Am: iPro: Wa (10: 4: 5) = 0.33 on SiO ₃ .

B. Boc-Arg (NO ₂ ) -Tra

5.76 g of Boc-Arg (NO ₂ ) -OH were dissolved in 70 cm ^{5 of} DMF and 2.52 cm ^{5 of} TAA. This solution was cooled to -10 C and then 2.38 cm isobutyl chloroformate was added and the mixture stirred for 10 min at -10 ⁰ C. To this solution was added a solution of 2.9 g of tryptamine in 10 cm ⁵ cm ⁵ DMF and 2.32 TAA added, the temperature was maintained at about -1O ⁰ C.

The mixture was stirred for 30 min at -10 ⁰ C ₁ 2 h at ⁰ C and 18 h at 20 ⁰ C and then the solvent is evaporated under vacuum and the ftückstand in ethyl acetate / water was added.

- 37 309836/1 185

The organic phase was washed with water, 5% bicarbonate solution and water, then dried and evaporated. Yield: 7.4 g of oil; Rf in Bz: EtOH (8: 2) = 0.54 on SiO ₃ .

G. Z-Phe-Arg (NO g ) -Tra and H-Phe-Arg-Tra

2.1 g of Z-Phe-ONP were in 10 oar ⁵ DMF and 10 oa? · Dissolved Äthylaoetat Then, 1.81 g of H-Arg (N0 ₂₎ -Tra (obtained by cleavage of the Amida of example, to the 2 G. described in example) was added, and then the Gemisoh 30 min at -1O ⁰ C and Stirred at 20 ^° C. for 20 h. The reaction mixture was processed further as described under B. Yield: 1.3 g; Pp 131-133 ⁰ C.
Rf in Bz: EtOH (8: 2) = 0.47 on SiO ₂ .

1.3 g of the compound obtained were converted into H-Phe-Arg-Tra by hydrogenation over ΙΟ? * Palladium on carbon. Yield:

D. H-Val-Glu-Hi s-Phe-Arg-Tra

The hydrazide prepared according to A. was converted into the azide in the usual way with the aid of isoamyl nitrite and coupled with the peptide H-Phe-Arg-tryptamide from stage C. The HeaJrtionsgemisch was stirred for 70 h at 0 ⁰ C, and poured into 0.1 η sodium bicarbonate solution, wherein the peptide is precipitated.

After filtering off the precipitate was dissolved in DMF and the solution was poured into ten times the amount of ethyl acetate. Filtering gave a residue with Rf in Bu: Py: Ao: Wa (4: 3/4: 1/4: 1) = 0.54 on SiO ₂ .

The above-described cleavage (Example 2 G.) gave the acetate of H-Val-Glu-His-Phe-Tra, yield: 33 $. Rf in the same mixed solvent = 0.18.

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Example 14 D-Met-Glu-Hia-Phe-Lys-tryptejiid derivatives

A. Z-Phe-Lys ( Boo ) -Tra

Auegehend prepared from 3.4 g of Z-Phe-ONP and 3 g of H-Lys (Boo) Tra of Z-Lys (Boc) -Tra (m.p. 77-80 ⁰ C) was Z-Phe-Lys (Boo ) -Tra obtained in the manner described in Example 13 G.

Yield: 65 #, mp 125-129 ⁰ C.
Hf in Bz: EtOH (8: 2) = 0.70 on SiO ₂

B. H-Phe-Lys (Bo ο) -Tra

2 g of the peptides described above (A.) were in 25 cm

Dissolved methanol and hydrogenated in the usual way in the presence of palladium on charcoal. After evaporation to Trookene, a foam was obtained, yield ^r rjfo. Rf in Bu: Ac: Wa (4: 1: 1) = 0.80

C. Boo-A-glu (0tBu) -Hl3-Phe ~ Ly9 (Boo) -Tra (A = Met, D-Met or

ß-Ala)

The tryptamide derivative of B. in DMF, 1.5 g and the hydrazide BoO-A-SIu (OtBu) -HiS-N ₂ H ^ were coupled according to the azide method described in the previous examples. After evaporation of the DMF, the residue was taken up in aqueous ethyl acetate and the organic phase was washed with water / water and 5% bicarbonate solution.

The organic phase was distilled off and the residue was recrystallized from ethyl acetate / alcohol / ether.

Peptide: Hf in Bz: BtOH (8: 2)

Boo-Met-Glu (OtBu) -His-Phe-Lys (Boc) -Tra 0.52 Boc-D-Met-Glu (OtBu) -His-Phe-Lys (Boo) -Tra 0.52 Boc-β-Ala-Glu (OtBu) -His-Phe-Lys (Boo) -Tra 0.49

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D. De8amino-Met-Glu (OtBu) ~ His-Phe-Lys (βoc) -Tra

In the manner described under C. ", the hydrazide desamino-Met-Glu (OtBu) -His-NpHp, which has been obtained from the corresponding methyl ester (2 B.), was coupled with H-Phe-Lys (Boc) -Tra (B.) using the azide method described above.
Rf in Bz: EtOH (8: 2) = 0.45

E. Removal of the protecting groups from the peptides obtained in C. and D.

In the manner described in Example 2 G., the peptides indicated above were removed from the protective groups with the aid of TFA freed and then exchanged the TFA of the peptide salt obtained in this way by acetic acid with the aid of a Ion exchanger in acetate form (e.g. Dowex Χ-Θ in acetate form)

Peptide: Rf in Bu: Py: Ac: Wa (4: 3/4: 1/4: 1)

on

H-Met-Glu-His-Phe-Lys-Tra - 0.20

H-D-Met-Glu-His-Phe-Lys-Tra 0.18

H-β-Ala-Glu-His-Phe-Lys-Tra 0.17

Desamino-Met-Glu-His-Phe-Lys-Tra 0.25

P. SuIfoxid e

45 mg of one of the peptide derivatives obtained under E. were dissolved in 2.5 cm acetic acid and then 15 JoOL 30% hydrogen peroxide was added. The mixture was treated in the manner described in Example 1. The residue was taken up in ßutanol / water (1: 2) and lyophilized.

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Peptide: Rf in Bu: Py: Ac: v / a (4: 3/4: 1/4: 1)

on SiO ₂

H-D-Met (-> O) -Glu-Hia-Phe-Lys-Tra 0.16

H-L-Met C * 0) -GIu-His-Phe-Lys-Tra 0.18

Desamino-Met (■ »0) -Glu-His-Phe-Lys-Tra 0.22

Example 15
Synthesis of HA-Glu-His-Phe-Arg-Trp derivatives

A. Boo-Arg (NO ₀ ) -Trp-ONB

5.76 g of Boc-Arg (NO ₂ ) -OH were dissolved in 70 cm ^{5 of} DMP and 2.52 cm ^{5 of} triethylamine. The solution was cooled to -10 ⁰ G and then 2.38 cm isobutyl chloroformate were added and the mixture stirred for 10 min at -10 ⁰ C. A solution of 6.14 g of H-Trp-ONB.HCl in 40 gto? cooled DMF and 3.01 cm ^{5 of} triethylamine were added.

The mixture was stirred for 30 min at -1O ⁰ C, 3 h at ⁰ C and 20 h at 20 ° C and then the solvent was evaporated in vacuo and the residue dissolved in ethyl acetate / water (1: 1) was added. The organic phase was washed with water, 5 # sodium bicarbonate solution and water, dried and evaporated to dryness in vacuo.
Yield: 8.5 g of oil.
Ef in Bz: EtOH (8: 2) = 0.79 (SiO ₉ ).

B. H-Arg (NO ₂ ) -Trp-ONB

1 g of the peptide prepared according to A. was dissolved in 20 cm of methylene chloride and then hydrogen chloride gas was passed through with cooling. The solution was stirred for 1 hour and then the precipitate was filtered off and washed thoroughly with dry methylene chloride. The precipitate was processed further immediately. Hf in Am: Py: Wa (5: 3: 2) = 0.66 on SiO ₂ (Rf starting S3 substance 0.93).

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C. Z-Phe-Arg (NO ₂ ) -Trp-ONB

3 g of the dipeptide prepared under B. were dissolved in 20 cm DMF. This solution was ^{cooled to 0} ° C. and then 1.12 cm ^{5 of} TAA and 2.12 g of Z-Phe-ONP were added. The reaction mixture was stirred for 2 h at ⁰ C and 20 h at 20 ⁰ C and evaporated in vacuo.

The oily residue was dissolved in ethyl acetate / V / water (2: 1) dissolved and processed as described under A.

The organic phase was evaporated, the residue in

3 3

Dissolve 5 om methanol and slowly add 100 cm of dry ether poured, whereby the peptide precipitated.

Yield: 2.9 g; Ä7J) ² = -15 ° (ο = 1, DMF); Rf in Bz: EtOH (8: 2) = 0.48 on SiO ₂ -

D. H-Phe-ArK-Trp-OH

2 g of the above-mentioned peptide C. were in 40 cm acetic acid dissolved and then 1 g of 10 ^ palladium on carbon was added and the mixture was hydrogenated for 2 days (Parr.).

After filtering, the acetic acid was evaporated off in vacuo and the residue was then stirred in dry ether. Yield 98 #. The light colored foam was dried over solid sodium hydroxide. The substance contained 1.1 moles of acetic acid.
Rf in Am: Py: Wa (5: 3: 2) = 0.17 on SiO ₃ .

E. Boo-D-Met-Glu (OtBu) -His-Phe-Arg-Trp-OH

1.17 g of the tripeptide Boc-D-Met-Glu (OtBu) -HiS-N ₀ H

3
(Example 9 A.) were dissolved in 20 cm DMF. After addition of 3 om ³ 2 η HCl in THP at ⁰ ° C the mixture was cooled to -2O ⁰ C. Then 0.27 cm ^{3 of} isoamyl nitrite was added. The Gemisoh was stirred for a few minutes and then this "azide" solution was added to 8 om DMF in which 2 mmol of the _ ^ ₂

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Tripeptides from D. (H-Phe-Arg-Trp-OH.aoetat) were dissolved. The pH of the mixture was then adjusted to 7.3 with TAA. The reaction mixture was ^{stirred at 0} ° C. for 70 h and then evaporated under reduced pressure and the kick stand was stirred in aqueous ethyl acetate.

The organic phase was then washed with water and then dried. After filtering the organic solution, petroleum ether was added to precipitate the peptide. Hf in Bu: Py: Ao: Wa (4: 3/4: 1/4: 1) = 0.52 on SiO ₂ .

P. Boo-Gly-Glu (OtBu) -His-Phe-Arg-Trp-OH

2 mmol of the tripeptide BoO-GIy-GIu (OtBu) -HiS-N ₂ H ₃ , prepared from Boc-Gly-Glu (OtBu) -His-OMe by conversion with hydrazine hydrate, were coupled to the peptide H-Phe-Arg- Trp-OH in the manner described under E. The reaction mixture was stirred for 70 h and then poured into 100 cm of ethyl acetate and extracted twice with salt-containing water.

The organic phase was evaporated to Trookene. The kickstand was stirred with dry ethyl acetate / potrol ether (9: 1) and dried. Ep 214-217 ° C (dec.). Hf in Bu: Py: Ao: Wa (4: 3/4: 1/4: 1) = 0.49 on SiO ₂ .

_x ) G. In the same way were prepared: Hf

1. Boo-Val-Glu (OtBu) -His-Phe-Arg-Trp-OH 0.49

2. Boo-β-Ala-Glu (OtBu) -His-Phe-Arg-Trp-OH 0.48

3. Boo-Ala-Olu (0tBu) -His-Phe-Arg-Trp-0H 0.52

4. Boo-feC-Me) Ala-Glu (OtBu) -His-Phe-Arg-Trp-OH 0.54

5. Deeamino-Met-Glu (OtBu) -His-Phe-Arg-Trp-OH 0.48

*) Hf in Bu: Py: Ao: Wa (4: 3/4: 1/4: 1) on SiO ₉ .

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H. The peptides described under E., P. and G. were on Freed from the protective groups in the manner described in Example 2 G. The acetates of the following peptides were obtained.

Rf *)

1. H-Val-Glu-His-Phe-Arg-Trp-OH 0.20

2. H-Gly-Glu-His-Phe-Arg-Trp-OH 0.18

3. H-β-Ala-Glu-His-Phe-Arg-Trp-OH 0.19

4. H-D-Met-Glu-His-Phe-Arg-Trp-OH 0.19

5. H-Ala-Glu-His-Phe-Arg-Trp-OH 0.20

6. H-K-Me) Ala-Glu-His-Phe-Arg-Trp-OH 0.21

7. Desamino-Met-Glu-His-Phe-Arg-Trp-OH 0.23

* 'Rf in Bu: Py: Ac: Wa (4: 3/4: 1/4: 1) on SiO ₂ .

Example 16 Synthesis of D-Met-Glu-His-Phe-Lys-Trp

A. Z-Phe-Lys (Boo) -Trp-OH

2.1 g of Z-Phe-ONP-ester were coupled with 2.24 g of H-Lys (Boc) -Trp-OMe, as in Example 1? G. described. The oily ester was isolated and then saponified with 1.1 eq sodium hydroxide in 10 cm methanol. On acidification, the tripeptide acid precipitated and was recrystallized from methanol / water. Yield 2.61g.
Rf in Bz: EtOH (8: 2) = 0.23 (SiO ₂ ).

B. H-Phe-Lys (Boo) -T rp-OH

Hydrogenation of the tripeptide derivative of A. gave a foam in quantitative yield. Recrystallization from water / methanol gave the tripeptide in 71% yield.
Rf in Bz: EtOH (8: 2) = 0.05 (SiO ₂ ).

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C. R-Glu (OtBu) -HiS-PhB-LVa (Boc) -Trp-0H (R = Desaraino-Met or

Boc-D-Met)

In the manner described in Example 7, R-GIu (OtBu) -HiS-N ₂ H ₃ , where R denotes desamino-Met or Boc-D-Met ", was coupled to the tripeptide which had been prepared according to B. using the azide method. The DMF solution was poured into ten times the amount of water containing acetic acid (pH 3-4), whereby a precipitate was obtained to 60 ^ iger yield isolated.

D. R-GIu-His-Phe-Lys-Trp-OH (R = Desamino-Met or HD-Met)

Upon cleavage of the protected peptide which had been prepared according to C. to that described in Example 2 G. Way, one obtained the acetate of the corresponding unprotected peptide.

Rf in Bu: Py: Ac: Wa (4: 3/4: 1/4: 1) = 0.21 on SiO ₂ (for the HD- ^ iet peptide) and 0.25 (for the desamino-Met- Peptide).

E. Sulphoxide of R-GIu-His-Phe-Lys-Trp-OH (R = Desamino-Met

or H-D-Met)

In the manner described in Example 1, the peptides obtained under D. were oxidized with acetic acid containing 30 fo HpO _2.

Rf in Bu: Py: Ac: Wa (4: 3/4: 1/4: 1) = 0.19 on SiO ₂ for the D-Met (^ O) -peptide and 0.22 for the desamino-Met ( O) peptide.

Example 17

Soloxide of H-Met-Glu-His-Phe-Arg-Trp-OH and of H-Met-Glu-Hia-Phe-Lys-Trp-OH

By oxidation of Η-Met-GIu-His-Phe-Arg-Trp-OH and H-Met-Glu-His-Phe-Lys-Trp-OH with 30% hydrogen peroxide in acetic acid (see Example 1) were the corresponding

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Obtain sulfoxides, which were purified in countercurrent. Counter-atom system: Bu: Ac: Wa (4: 1: 5). Rf in Bu: Py: Ac: Va (4: 3/4: 1/4: 1) = 0.17 for the sulfoxide of the "Lys peptide" and 0.20 for the sulfoxide of the "Arg peptide".

Example 18 Synthesis of A-Glu-His-Phe-Arg-Trp-Gly peptides

A. Boo-D-Met-Glu (OtBu) -His-Phe-Arg-Trp-Gly-OH

In the manner described in Example 9, the

Boc-D-Met-Glu (OtBu) -HiS-N ₂ H, coupled with H-Phe-Arg-Trp-Gly-OH. Aoetat.

Rf in Bu: Py: Ac: Wa (4: 3/4: 1/4: 1) = 0.49 on SiO ₂ .

B. HD-Met-Glu-fiis-Phe-Arg-Trp-Gly-OH

When the protected peptide which had been obtained according to A. was cleaved in the manner described in Example 2 G., the acetate of HD-Met-GIu-His-Phe-Arg-Trp-GIy-OH was obtained.
Rf in Bu: Py: Ac: Wa (4: 3/4: 1/4: 1) = 0.16 on SiO ₂ .

C. Sulphoxide of HD-Met-Glu-His-Phe-Arg-Trp-Gly-OH

The peptide prepared according to B * was oxidized with the aid of acetic acid and 30% H ₂ O ₂ in the manner described in Example 1 and then lyophilized. Rf in Bu: Py: Ao: Wa (4: 3/4: 1/4: 1) = 0.14 on SiO ₂ -

D. In the same manner, the following peptides were prepared.

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H-Gly-Glu-His-Phe-Arg-Trp-Gly-OH H-ß-Ala-Glu-His-Phe-Arg-Trp-Gly-OH H- (oC-Μθ) Ala-Glu-Hi s-Phe-Arg-Trp-Gly-OH H-Val-Glu-His-Phe-Arg-Trp-Gly-OH Deeamino-Met-Glu-Hia-Phe-Arg-Trp-Gly-OH Deaamino-Met (- ♦ O) -GIu-Hi s-Phe-Arg-Trp-Gly-OH.

Example 19 Sulphoxide of H-Het-Glu-His-Phe-Arg-Trp-Gly-OH

In the manner described in Example 1 ", the heptapeptide H-Met-Glu-His-Phe-Arg-Trp-Gly-OH is oxidized and then lyophilized. Rf in Bu:? Y: Ao: Wa (4: 3/4: 1/4: 1) = 0.16

Example 20 SuIfon from H-Met-Glu-His-Phe-Lys-Trp-Gly-OH.aoetat

200 mg of the peptide H-Met-Glu-His-Phe-Lys-Trp-Gly-OH were in a mixture of 0.5 cm water, 0.1 cm 4 η perchloric acid,

• 5

0.02 cm 0.5 m ammonium molybdate were dissolved and then • x

0.06 om 30% hydrogen peroxide added.

The Gemisoh was stirred for 2 h at a temperature of about 1O ⁰ C. The mixture was then stirred with Dowex X-8 in the acetate form and then filtered and the Piltrat ™ lyophilized. Rf in Bu: Ac: Wa (4: 1: 1) = 0.21

In the same way were prepared: Η-Met (4O ₂ ) -Glu-His-Phe-Lys-Tra Deaamino-Met (-> 0 ₂ ) -Glu-His-Phe-Lys-Trp-0H Desamino-Met (- ^ O ₂ ) -GIu-Hi s-Phe-OH.

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Example 21 Preparation of Η-Met (d, - »O) -Glu-His-Phe-Arg-Trp-Gly-OH

A. Boo-Met (d, - »0) -OSu

26 g of L-methionine were oxidized to the corresponding sulforoxide according to the method of Toennies and KoIb cf. Biol. Chem. 128, 399 (1939). 24 g, pp 250 ° C (decomp.) The two diastereomeric sulfoxides were separated according to the method of Lavine, J. Biol. Chem. 169, 477 (1947). Yield: L-methionine (d, - »0) 8.7 g Z ° L7n + 110 ° (o = 1, L-methionine (1, - ^ 0) 1.8 g / j * J ²² -75 ° ( o = 1,

These isomers were then converted into the Boc-protected derivatives using Boc-N-, in the usual way: Boc-Met (d, -> O) -OH Pp 135-137 ° C / ^ + 42 ° (c = 1, DMP) Boc-Met (1 -? + O) -OH mp 68 ⁰ C / i7u ² "58 ° (ο = 1, DMP) and then to the corresponding N-hydroxysuccinimide ester by treatment with DCCI and HOSu this. active ester derivatives were then used immediately for the following coupling reaction.

B. H-Met (d, -> 0) -GIu-Hi s-Phe-Arg-Trp-Gly-OH

929 mg of Boc-Met (d-sulfoxide) -0Su were dissolved in 20 cm ^{5 of} DMP. The solution was ^{cooled to 0} ° C. and then a solution of 2.38 g of H-Glu (0tBu) -His-Phe-Arg-Trp-Gly-OH and 0.83 cm of N-ethylmorpholine in 25 cm of DMP was added.

The reaction mixture was ^{stirred for 3 h at 0} ° C., 70 h at room temperature and then evaporated. The deficit

• 2,. it.

was stirred with 40 cnr ethyl acetate and 10 cm water, filtered and washed with petroleum ether and dried. Yield: 2.1 g; Rf 0.44 (Bu: Py: Ac: Wa - 4: 3/4: 1/4: 1)

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2 g of Boc-Met (d-sulfoxide) -Glu (OtBu) -His-Phe-Arg-Trp-Gly-OH were treated with 20 cnr 90 tiger TFA for 30 min. The solution was then stirred in 300 cm of ether, whereupon the precipitate which separated out was filtered off, washed with ether and dried over solid Potassium hydroxide was dried. The trifluoroacetate was in 50 cm tert-butanol / water (1: 1) dissolved and treated converted into the acetate with Dowex X-8 in the acetate form. After filtering, the filtrate was lyophilized. Yield: 1.45 g; Rf 0.24 in Bu: Py: Ac: Wa (2: 3/4: 1/4: 1)

Claims

^62XXI 309836/1185

Claims (5)

DR. ING. F. -WTJKSTHOFF 8 MUNCH EN ΘΟ DR.JS.T.P1ICHMAHN aCHWKIOKBSTRASSE DKiING. DiBKHItBNS 2308362 ""'°"*"" ^M """ DIPL ·. ING. H- GOETZ PATBKTAKWZLTK Il ft hotmtmtmt MS. U-42 571 Claims Peychopharmacologically active peptides and peptide derivatives of the formula: A-I-GIu-L-His-X, in which - A-H-D-Met, H-Ir-Met £ 0), H-D-Met (-K)), HL-Met (- * 0 ₂ ) # HD-Met (- ^ Op) ₁ Desamino-Met, Desamino-Met- (-τ O), Desamino-Met (- »O ₂ ) or the group H ₂ NB-CO -, in which B is a branched or unbranched alkylene group with 1 to 6 carbon atoms, X is a hydroxyl group, an N-phenyl alk ylamino group of the general formula -HH - Alk - ^ ~ ^ C ^R 1 ic _{or R 1 is} a hydrogen or Halogen atom or a hydroxyl group, an alkyl group with 1 to 4 carbon atoms or an alkoxy group with 1 to 4 carbon atoms and Alk a branched or unbranched alkylene group with 1 to 6 carbon atoms ISt _x or the group L-Phe-Y in which Y is a hydroxyl group , an (N-Aminoalky]) emiono group of the general formula -NH-AIk-NR ₂ IU in which R _{2 is} a hydrogen atom or a lower alkyl group with 1 to 6 carbon atoms, R ^ a * 'hydrogen atom, a lower alkyl group with 1 to 6 Carbon atoms or an amidine group and Alk is a branched or unbranched alkylene group having 2 to 6 carbon atoms, or the group L-Lys-Z or L-Arg-Z, in which Z is a hydroxyl group, the group L-Trp-OH, L-Trp-Gly-OH or a (N-3-indolylalkyl) aniino group, the alkyl group thereof Contains 1 to 6 carbon atoms 309836/1 185 as well as the functional derivatives of these peptides and Peptide derivatives, 2. Peptides according to claim 1, characterized in that that A is a ß-alanyl group or a sulfoxide or Sulphone group of the methionyl or desaminomefchionyl group and X has the meaning given above. 3. Peptide according to claim 1, characterized in that that it has the general formula A-L-GIu-L-His-L-Phe-L-Arg- (or L-Lys) -Tra, in which A has the meaning given above has and Tra means a tryptaraid group, owns as well its functional derivatives. 4. Peptides according to claim 1 to 3, characterized in that g e k e η η draw *, that they are present in ports of the metal complexes. 5. Process for the production of the peptides according to claims 1 to 4, this shows that the individual Peptide -Fregniente in the correct order couples and then-so / eit available- protective groups removed and the peptides thus obtained optionally converted into the functional derivatives and / or into the corresponding peptide sulfoxides or peptide sulfones transferred if the remainder of the kit (terminal) nitrogen atom is a Met .- or Desaminomet group. 62X1 309836/1185