HU176260B - Process for producing encephalin-analogue polypeptides - Google Patents

Abstract

The present invention relates to a process for the preparation of polypeptide derivatives of general formula I. and their pharmaceutically acceptable addition salts, characterized in that j- '.' .. '. a) one or more conventional peptide protecting groups are removed from a protected polypeptide to give the compound of formula I, or ', .... b) a carboxylic acid of formula II or its activated derivative with a suitable one Alcohol or amine is reacted. The compounds produced by the process according to the invention have analgesic properties, formulas I and II

Description

Survived by Anand Swaroop Chemist, Gormley James Joseph Chemist, Haywaid Christopher Frederick Chemist, Morley John Selwyn Chemist, Stacey Gilbert Joseph Chemist, Macclesfield, Cheshire, UK

patentee:

Imperial Chemical Industries Limited, London, UK

Process for the preparation of encephalin analog polypeptides

2

The present invention relates to novel enkephalin analog polypeptide derivatives having analgesic properties.

In Nature 258, 577-579 (1975) are two related, called "enkephalins" for ⁵ discloses penta- peptide. These compounds were isolated from porcine brain. The compounds have the formula H-Thyr-Gly-Gly-Phe-Met-OH, respectively

H-Tyr-Gly-Gly-Phe-Leu-OH. Experiments have shown that these compounds exert opium-like activity on isolated organ preparations. Although these two compounds, when injected into the cerebral cortex of cats, have a transient analgesic effect, they have been shown to be ineffective in conventional analgesic assays in laboratory animals.

It has been found that modifying the amino acid structure of the enkephalin molecule results in compounds which exhibit good analgesic activity even when administered intravenously.

The enkephalin analog polypeptide derivatives obtainable by the process of the present invention have the general formula (I) wherein (i)

R ¹ is hydrogen or methyl,

R ² is hydrogen or acetyl, or Glu, Asp, δ-Ala, D-Lys, Lys, H-Glu-Gly-OH optionally substituted on the ε-hydroxy carbonyl group by t-butoxycarbonyl. H-Lys-, * · {------------- H

H-Phe Η-Lys-, H-Gly-Gly-Gly H-Lys-,

H-Asp Η-Lys-, H-Lys H-Lys-,

H-Gly-Gly, H-Leu-Leu-Leu, H-Arg-ProLys, H-Pro-Gin-Gin, H-Gly-Gly-Gly, H-Lys-Gly-Gly-Gly, H -Asp-Gly-Gly-Gly, H-Arg-Pro-Lys-Pro-Gln-Gln, or H-Gly-Gly-Gly-Gly-Gly-Gly-,

B is D-Ala, O-Ala or Azala,

E is Gly or Azgly,

F is Phe, hexahydro Phe or Azphe, G is DL-Leu, Leu, D-Leu, Azleu, Nle, Met, Pro or Azpro, and

K is hydroxy, amino, phenoxy, C 1-6 alkoxy, C 2-4 alkenyloxy, a hydroxy, amino, C 1-4 alkylamino, N- (C 1-4) -alkyl-N-benzyloxycarbonylamino or C 1 -C 20 alkanoyloxy substituents or two C 1 -C 20 alkanoyloxy substituents, C 1 -C 4 alkoxy groups, C 1 -C 4 alkylamino groups, C 5 -C 7 alkoxyalkylamino group, di - (C 1 -C 4) -alkylamino, hydroxy, (C 1 -C 4) -alkylamino or (C 1 -C 4) -alkylamino substituted with (C 1 -C 4) -alkylamino, morpholino- a tetrahydropyrrolo group or a (1-4C) alkyl group

G and K together form a group of formula (III) or (IV) in the D, L · or DL configuration, or a group of formula (II), provided that at least one of B, E, F and G is represents a group other than the aza-amino acid residue, or (ü)

R ¹ and R ² are both hydrogen,

K represents a C 1-6 alkoxy group,

G is Leu, D-Leu, Azleu or Nle,

E is Gly or Azgly,

F is Phe, hexahydroPhe or Azphe, 15 and

B is D-Asp, D-Thr, D-Trp, D-Leu, D-Lys or D-Met, with the proviso that at least one of E, F and G is other than an aza-amino acid residue, or 20 (ui)

R ^{1 is} hydrogen,

R ² is hydrogen, D-Lys, or Lys optionally substituted on the amino group at the ε-position by a t-butoxycarbonyl group,

B is a D-Ser group optionally substituted on the hydroxy group at the N-position by a t-butyl group,

K is C 1-6 alkoxy, 3θ

E is Gly or Azgly,

F is Phe, hexahydroPhe or Azphe, and

G is a Met group, provided that at least one of E and F is other than 35 aza-amino acid residues, or (iv)

R ^{1 and} R ² are each hydrogen,

K is an amino group optionally substituted with one or two C 1-4 C 49 alkyl groups,

B is selected from the β-hydroxyl group substituted with D-Ser group is a t-butyl group, 4 ₅

E is Gly,

F is Phe, and

G is Pro or Azpro, or (v)

R ¹ is hydrogen or methyl,

E Gly means a group,

F is Phe, hexahydroPhe or Azphe,

G is Leu, D — Leu, Azleu, or Nle,

Λ 55

R is hydrogen, D-Lys, Lys optionally substituted on the amino group at the e-position, or H-Gly-Gly-Gly,

K represents a hydroxy group, an amino group, a C 1 -C 6 C 60 alkoxy group, a C 1 -C 4 alkoxy group substituted by a hydroxy or C 1 -C 20 alkanoyloxy group or two C 1 -C 20 alkanoyloxy groups or a tetrahydropyrrolo group; group 65, and

B is a D-Ser group optionally substituted on the hydroxy group at the 0-position by a t-butyl group, or (vi)

R ¹ is hydrogen or methyl, R ² is hydrogen or acetyl, B is D-Ala or β-Ala, E is Gly,

F is Phe or hexahydroPhe, G is Leu or D-Leu, and

K is Gly-OMe, Thr-OH, D-Thr-OH, Ala-OMe or D-Al-OMe, and the amino acids listed are, unless otherwise indicated, in the L configuration.

The process of the present invention also provides pharmaceutically acceptable acid addition salts of the polypeptides of formula (I) containing the basic groups and the pharmaceutically acceptable base addition salts of the polypeptides of formula (I) containing the acidic groups.

In the specification and claims, amino acid residues are designated by the usual abbreviations (see Pure and Applied Chemistry 40, 317-331 (1974)). α-Aza-amniic acid derivatives are those amino acids in which the α-CH 1 group is replaced by a nitrogen atom. The a-aza-amino acid derivatives are distinguished from the parent compounds by the designation "Az". For example, Azala for aza-alanine, Azgly for aza-glycine, Azphe for aza-phenylalanine, Azleu for aza-phenylalanine, Azleu for aza-leucine and Azpro for aza-proline. The terms "hexahydroPhe" and "Phe (6H)" refer to the phenylalanyl group containing a cyclohexate ring at the place of the benzene ring. Unless indicated separately, the configuration of amino acids is to be understood as meaning any naturally occurring L-configuration amino acid. Of course, this convention does not apply to glycine with a symmetric structure and to those α-aza-amino acids which do not contain an asymmetric carbon atom adjacent to the carboxyl group.

Particularly preferred are those of claims 1 to 93. Compounds of formula (I) prepared according to example 1 to 4, and in particular the following:

H-Tyr-D-Ala-Gly-Phe-DL-NH- (2-oxo-tetrahydrofuran-3-yl);

H-Tyr-D-Ala-Gly-Phe-Pro-NHC ₂ H _s , H-Tyr-D-Ser-Gly-Phe-Azpro-NHC ₂ H _s ,

H-Leu-Leu-Leu-Tyr-D-Ala-Gly-Phe-Leu-OCH ₃₎ H-Gly-Gly-Gly-Tyr-D-Ala-Gly-Phe-Leu-OCH ₃ , H-Tyr- D-Ala-Gly-Phe-Leu-NH (CH ₂ ) ₂ NHCH ₃ , H-Tyr-D-Ala-Gly-Phe-DL-Leu-CH ₃ ,

H-Tyr-D-Ser-Gly-Phe (tetrahydropyrazolo)

H-Tyr-D-Ser-Gly-Phe-Pro-NHC ₂ H _s , H-Lys-Tyr-D-Ala-Gly-Phe-DL-NH- (2-oxo-tetrahydrofur-3-fl),

H-Tyr-D-Ala-Azgly-Phe-Azleu-NH ₂ ,

H-Lys-Tyr-D-Ala-Gly-Phe-Azleu-NH ₂ , H-Tyr-D-Ala-Gly-Phe-Leu-D-Thr-OH,

H-Tyr-D-Ala-Gly-Phe-Leu-NIXCH ₂ ) ₂ N (CH ₃ ) ₂ , H-Tyr-D-Ser-Gly-Phe-Leu-OCH ₃ .

Examples of acid addition salts of the compounds of formula I include hydrochlorides, phosphates, citrates, acetates and trifluoroacetates.

Exemplary base addition salts of the compounds of formula I include ammonium salts.

The polypeptide derivatives of formula (I) may be prepared by methods known per se. In the preparation of these compounds

(a) cleavage of protecting groups of the corresponding protected polypeptides; or

(b) in the preparation of compounds of formula (I) wherein K is other than a hydroxyl group, and in the preparation of compounds of formula (I) in which G and K together form a compound of formula (II): a carboxylic acid of the general formula wherein R ¹ , R ² , B, E, F and G are as defined above or a reactive derivative thereof with the appropriate alcohol or amine.

The polypeptides used as starting material in process variant a) may carry protecting groups on protecting groups (for example, the free —OH or —NH groups of the final product). These protecting groups may be those known and commonly used in peptide chemistry (see, e.g., Bodansky and Ondetti, "Peptide Synthesis", Interscience, New York, 1966, IV. Chapter II, "The Proteins" by F.M. Finn and K. Kaufmann. Vol. 106, published by H. Neurath and R. L. Hill, Academic Press Inc., New York (1976), p. 106; and " Amino acids, Paptides and Proteins ", Specialist Periodical Reports, The Chemical Society, London, 1— Volume 8] The cited literature describes several methods for deprotection.

Benzyloxycarbonyl is preferably used to protect the amino group, and benzyl is preferably used to protect the hydroxy group. Both protecting groups are easily removed by hydrogenolysis. Hydrogenolysis may be carried out in the presence of, for example, palladium on charcoal (such as palladium on charcoal containing 5% by weight of palladium) using a diluent or solvent such as water, methanol, ethanol, butanol, dimethylformamide, acetic acid, chloroform or a mixture of these. The reaction may optionally be carried out in the presence of an acid such as hydrochloric acid or p-toluenesulfonic acid. The reaction is usually carried out at atmospheric pressure.

In addition, tert-butoxycarbonyl is preferably used for protecting the amino group, and tert-butyl for protecting the hydroxyl group. Both protecting groups can be readily cleaved by treatment with acid (e.g. hydrochloric acid or trifluoroacetic acid). Hydrochloric acid may be introduced into the system in the form of an aqueous solution (e. G., A 1 N to saturated aqueous solution) or mixed with other diluents or solvents such as ethyl acetate, methanol, acetic acid, ether and / or dioxane. The concentration of hydrochloric acid solutions in organic solvents may be up to saturation, but solutions of 2 to 6 n are preferred. The reaction is preferably carried out at a temperature between 0 ° C and room temperature. Optionally, the reaction may be carried out in the presence of 5 cleavers, such as 2-mercaptoethanol. Trifluoroacetic acid can be added undiluted or diluted with 5-10% water. The reaction is preferably carried out at room temperature, optionally in the presence of a cleavage agent such as 2-mercaptoethanol.

In addition, benzyloxycarbonyl or tert-butoxycarbonyl may be advantageously used to protect the amino group when tert-butyl is used to protect the hydroxy group. These protecting groups can be cleaved with acetic acid hydrobromide. The concentration of the hydrogen bromide solution in acetic acid is preferably 1-3 n, preferably 2 n.

The carboxyl group is preferably protected as an ester such as methyl or ethyl esters ^20. These protective groups can be split off by base hydrolysis, for example, the protected compound of solvent or diluent such as aqueous methanol, aqueous cellosolve, treated with sodium hydroxide ²⁵ or potassium hydroxide in aqueous dioxane or aqueous dimethyl formamide. The reaction is generally carried out at room temperature.

Process variant b) may use, for example, the corresponding esters or acid anhydrides as reactive derivatives of the carboxylic acids of formulas V and VI. Starting from reactive carboxylic acid derivatives, these compounds are reacted with appropriate alcohols or amines in the presence of a diluent or solvent. When starting from the free carboxylic acids of formula (V) or (VI) 35, the reaction is preferably carried out in the presence of a known peptide coupling agent such as Ν, Ν'-dicyclohexylcarbodiimide. This process is particularly useful for the preparation of compounds of formula I wherein K is an amino group or a substituted amino group. In this case, the esters of the compounds of formula V, for example the corresponding methyl ester, are reacted with excess ammonia or the corresponding substituted amine in a solvent such as methanol or dimethylformamide at a temperature between 0 ° C and 45 room temperature. The reaction takes up to 3 days.

The starting materials 50 used in the process of the invention may be prepared from known compounds by known peptide chemistry (coupling, protecting group cleavage). The preparation of starting materials is described in Example A starting from the appropriate amino acids or protected amino acids. General methods for preparing starting materials 55 are described in Helv. Chim. Acta 62, 398-411 (1979) and J. Med. Chem. 21, 1110-1116 (1978). The same publications also describe the synthesis of some of the basic chains present in the structure of the compounds of formula I and the physical properties of the resulting peptides. No. 175 Intermediate is prepared by reacting the corresponding acid with acetic anhydride under pyridine conditions under Dakin-West conditions.

As previously reported, the novel compounds of general formula (I) and their salts exert analgesic activity on warm blood. The analgesic activity of the compounds was investigated in mice according to the method of Eddy and Leimbach [J. Pharmac. Exp. Therap. 107: 385-393 (1953)]. The tests were performed as follows:

Female mice weighing 22-25 g were included in the experiments. Three animals were used for each experiment. Mice were placed on a 56 ° C copper plate (the copper plate was thermostatically controlled) and the time when animals reacted to heat (for example, licking their hind legs) was recorded. The animals usually responded to the external stimulus after 3-5 seconds.

The mice were then injected intravenously with 100 mg / kg of the test compound. At 5, 10 and 30 minutes after administration, the mice were placed on the heated copper plate again and the time to reaction was measured. If the animal did not respond to the external stimulus after 20 seconds, the animal was lifted from the copper plate. This circumstance indicates that the compound exerts its maximum effect at the dose tested.

Compounds that prolonged the animals' reaction time by an average of at least 3 seconds were considered to be effective. Compounds that have been shown to be effective were re-tested at lower doses.

All of the compounds of formula (I) listed in the Examples were effective in the above experiment at doses of 100 mg / kg or less. The compounds in mice, LD ₅ o determined after intravenous administration significantly exceeding the range of the maximum analgesic effect triggering dose value. The results of some experiments are summarized in Table A).

The table

agent Reaction elongation, seconds ₅₀ mg / kg Example no. Formula Dose in mg / kg 49th H-Tyr-D-Ala-Gly-Phe-Leu (tetrahydro) 100 15 256 52nd H-Tyr-D-Ala-Gly-Phe-Pro-NC ₂ H _s 20 16 245 78th H-Tyr-D-Ala-Gly-Phe-Leu-NH (CH ₂ ) ₂ NHCH ₃ 10 15 90 58th H-Tyr-D-Ser-Gly-Phe-Met-OCH ₃ 50 15 464 34th H-Gly-Gly-Gly-Tyr-D-Ala-Gly-Phe-Leu-OCH ₃ 100 14 385

The compounds of the formula I and their salts can be converted into pharmaceutical compositions using conventional non-toxic pharmaceutical diluents, carriers and / or excipients. 45

The pharmaceutical compositions may be, for example, parenteral, sterile, aqueous or oily solutions or suspensions. These compositions are prepared in a known manner.

The pharmaceutical compositions include, in addition to the novel polypeptide derivatives, one or more other analgesic agents such as aspirin, paracetamol, phenacetin, codeine, pethidine or morphine, antiinflammatory agents such as naproxen, indomethacin and ibuprofen, a neuroleptic agent such as prochlorlorperazine, trifluorperazine and haloperidol, or other sedative or tranquillising agents such as chlorodiazepoxide, phenobarbitone and amylobarbitone.

Particularly preferred are pharmaceutical formulations for intravenous, intramuscular or subcutaneous injection, e.g.

Sterile aqueous solutions containing from 1 to 50 mg / ml of active ingredient.

Pharmaceutical compositions containing the novel polypeptide derivatives can generally be used in human medicine for the prevention or reduction of pain in the form of 2 to 150 mg of active ingredient by intramuscular or subcutaneous injection or 1 to 100 mg of active ingredient by intravenous injection.

The pharmaceutical compositions may be administered at intervals corresponding to the biological half-life of the active ingredient (e.g., 0.5 to 4 times the biological half-life). For example, the compositions may be administered two to six times daily. The novel formulations are particularly useful for reducing pain following surgical intervention. In this case, the compositions are preferably administered to patients during and shortly after surgery.

Injectable compositions may also be administered at intermittent doses, either by injection or by periodically adding an active ingredient to the infusion fluid in an infused patient. Alternatively, the active compounds may be administered by slow intravenous infusion.

The following examples illustrate the invention without limiting it.

i76260

The Rf values in the examples were determined by running up on a silica gel thin layer (Kieselgel G). The following solvent systems were used:

The system: Butane-1-ol: 4: 1: 5: acetic acid: water, System B: 15: 3: 12: 10 by volume butan-1-ol: acetic acid: water: : pyridine mixture, 10 System C: Butane-2-ol: 3: 1 by volume: 3% v / v aqueous ammonium hydroxide solution, System D: 3: 1 by volume aceto- nitrile: water mixture, 15 System F: Chloroform: ethanol (1: 4 by volume), System E): 1: 1 by volume acetone:: chloroform, System G): 1: 1 by volume cyclohexane:: ethyl acetate, 20 System H: 1: 1: 1 by volume mixture of cyclohexane: ethyl acetate: methanol, 25 System K): Chloroform: methanol: water (11: 8: 2 by volume), System P): 19: 1 by volume chloroform:: methanol, Q) system: Chloroform: methanol 9: 1 (v / v). 30 (G), K), Q) and P) on the silica gel column

The chromatographic plates were subjected to ultraviolet light, using fluorescamine, ninhydrin and chlorine starch iodine as the developing agent. Unless otherwise stated, Rf also means that only one spot is visible on the chromatogram. ⁴⁰

1-9. The acid peptide or protected peptide was heated in a sealed, evacuated tube at 110 ° C for 16 hours in acidic hydrolysis of the compounds described in Examples 1 to 6. The amino acid composition of the resulting hydrolyzates was determined using a LoCarts Amino-Acid Analyzer. In all cases, the detected amino acid compositions were consistent with the expected structure.

Abbreviations used in the examples have the meanings a

following: 50 Me - methyl, Bu ¹ = tert-butyl, Ac = acetyl, et = ethyl group, 55 Ph = phenyl, Z = benzyloxycarbonyl-c $ Bzl = benzyl, OCP = 2,4,5-trichlorophenoxy, Boc = tert-butoxycarbonyl, 60 ONp = p-nitrophenoxy, ONSu = succinimidooxy, toluene sulfonic = p-toluenesulfonic acid, DMF = dimethylformamide, TFA = trifluoroacetic acid. 65

Example A)

1-93 in this example. Examples include the preparation of starting materials for the preparation of the compounds of formula (I) listed in Examples 1 to 5. The preparation of the starting materials is summarized in tabular form. All of the starting materials are designated by Arabic numerals and are used in the preparation of further starting materials and final products of formula (I).

In the tables, the reactions leading to the formation of each starting material are indicated by letters and Arabic numerals. H1-H13. Methods 1 to 42 Example E1-E10. Methods 43-69 are the same. in example FI-F3. and method 70-88 is the same. . Further methods are described below.

Active ester reactions:

Method A1: A solution of the amino compound (1 mmol) and the active ester (1.1 mmol) in DMF (minimum volume) was maintained at 20-25 ° C. When the reaction mixture no longer reacts positively with ninhydrin (after 15-50 hours), the mixture is concentrated in vacuo to a volume which allows separation of the crude product. The crude product is isolated by precipitation with an organic solvent, by treatment with water or by treatment with a mixture of water and a suitable organic solvent (e.g. ether). If necessary, the crude product is further purified as indicated in the tables.

THE 2. Method A: Method A1 was repeated except that ethyl acetate was used as the reaction medium and the reaction was carried out at 4 ° C.

THE 3. Method A: Method A1, except that about 0.2 mmol of 1-hydroxybenzotriazole was added.

A4. Method A: Method A1 was repeated, but the reaction was carried out at 4 ° C.

A5. Method A: Method A1, except that about 0.2 mmol of 1-hydroxybenzotriazole was added and the mixture was kept at 4 ° C.

Mixed anhydride reactions:

Method B1: A solution of 1 mmol of the carboxylic compound in about 5 ml of dimethylformanide was cooled to -20 ° C to -40 ° C and 1.05 mmol of N-methylmorpholine was added followed by 1.05 mmol of chloroformic acid isobutyl ester. . The mixture was stirred at -20 ° C to -40 ° C for 5 minutes and then a solution of the amino compound in dimethylformamide (1 mmol) was added. When the amino compound is used as a salt, 1 mmol of N-methylmorpholine is added to the reaction mixture. The mixture was stirred at 20-25 ° C (24 hours maximum) until the reaction was complete and concentrated in vacuo. The residue is partitioned between dilute aqueous citric acid and a suitable water immiscible organic solvent (usually ethyl acetate). The organic phase was separated, washed successively with dilute aqueous nitric acid and aqueous sodium bicarbonate solution, dried and the solvent was evaporated. In the absence of a suitable organic solvent, a

the solid precipitated is washed (water is used as the last wash) and filtered.

B2. Method B: Method B1, but using triethylamine as the tertiary base.

B3. Method A: Method B1, but employing ethyl chloroformate to form the mixed anhydride.

B4. However, the mixed anhydride is formed in ethyl tetrahydrofuran using ethyl chloroformate and N-methylmorpholine.

B5. Method A: Method B1, however, the mixed anhydride is formed with ethyl chloroformate and triethylamine is used as the tertiary base.

D2. method Dl. However, 2 mmol of 1-hydroxybenzotriazole were added to the reaction mixture.

D3. Method 1: To a solution of 1 mmol of the carboxylic compound in 2 ml of anhydrous tetrahydrofuran was added 1 mmol of N-hydroxysuccinimide followed by 1 mmol of Ν, Ν'-dicyclohexylcarbodiimide in 1 ml of tetrahydrofuran. After 15 minutes, 1 mmol of the amino compound was added to the reaction mixture and the mixture was stirred for 15-25 hours at 20-25 ° C. The mixture is filtered, the filtrate is concentrated in vacuo and the residue is dissolved in a suitable solvent such as ethyl acetate. The organic solution was washed sequentially with aqueous citric acid solution, aqueous sodium bicarbonate solution, water, dried and concentrated in vacuo.

Azide reactions:

Method Cl: To a solution of 1 mmol of carboxylic acid hydrazide in about 5 ml of dimethylformamide at -20 ° C was added a 6 molar solution of 5-6 mmol of hydrochloric acid in dioxane, followed by the addition of 1.2 mmol of tert-butyl nitrite. The reaction of the hydrazide is examined by periodically sampling the sample, dropping the sample onto filter paper and examining it with a mixture of ferric chloride and potassium ferricyanide. After completion of the reaction, triethylamine (5-6 mmol) was added to the mixture at -10 ° C under stirring for 10 minutes, followed by addition of a solution of the amino compound (1 mmol) in DMF (3 mL). When the amino compound is used as its salt, an equivalent amount of triethylamine (1 mmol) is added to the reaction mixture. The reaction mixture was stirred at 0-4 ° C for 18-24 hours, then filtered and the filtrate was concentrated in vacuo. The residue is dissolved, if possible, in a water immiscible solvent, washed successively with aqueous citric acid solution, aqueous sodium bicarbonate solution and finally water, and the product is isolated by evaporation or filtration.

Ν, Ν'-Dicyclohexylcarbodiimide reactions:

Dl. Method 1: To a solution of 1 mmol of the carboxyl compound in 5 ml of dimethylformamide at 4 ° C was added 1.1 mmol of N, N'-dicyclohexylcarbodiimide and 1 mmol of the amino compound. The reaction mixture is stirred at 4 ° C (in some cases 20-25 ° C) until a positive ninhydrin reaction is no longer detectable (reaction requires up to 18 hours). When the amino compound is used as its salt, 1 mmol of triethylamine is added to the reaction mixture upon initiation of the reaction. The mixture is filtered, the filtrate is concentrated in vacuo, and the residue is dissolved in a suitable organic solvent such as ethyl acetate and washed successively with aqueous citric acid, aqueous sodium bicarbonate and water. The organic solution is dried and evaporated, or the product is precipitated from the solution with a suitable precipitant, such as petroleum ether (b.p. 60-80 ° C).

reduction:

Method G1: In preparing hexahydro-phenylalanine derivatives, the corresponding phenylalanyl-containing compound (1 mmol) is dissolved in 80% aqueous acetic acid. Optionally, a small amount of hydrochloric acid is added to the solution. To the reaction mixture is added 50-150 mg of Adams Platinum Oxide Catalyst, following standard precautions, and the reaction mixture is stirred under stirring at 20-25 ° C until the reaction is complete (15-50 hours). The catalyst was filtered off over diatomaceous earth and the filtrate was concentrated in vacuo. The crude product is obtained.

Hydrolysis of esters:

Jl. Method 1: Dissolve 1 mmol of the ester in 75% (v / v) aqueous acetone (1.1-1.2 mmol) containing sodium hydroxide (10-15 mL) and stir at 20-25 ° C. The reaction was followed by thin layer chromatography. After completion of the reaction (2-3 hours), most of the acetone was evaporated in vacuo and the resulting aqueous solution was washed with ethyl acetate. The aqueous solution is, for example, acidified with citric acid and the product is extracted with a suitable solvent such as ethyl acetate. If this route is not feasible, the product is isolated by evaporation. The extract was washed with saturated brine, dried, and concentrated in vacuo. The corresponding carboxylic compound is obtained.

J2. Method Jl. but the reaction medium is a mixture of water, methanol and acetone.

J3. Method Jl. but the reaction medium is aqueous methanol.

J4. Method 1: Hydrolysis is carried out with 1 molar equivalent of aqueous sodium hydroxide solution and the solution is evaporated. The product is isolated as its sodium salt.

J5. Method Jl. however, aqueous dioxane is used as the reaction medium.

Acylation reactions:

WHO. Method 1: Dissolve 1 mmol of the appropriate glyceryl monoester in 1 ml of chloroform containing 3 mmol of pyridine and add 3 mmol of acetyl chloride to the ice-cold solution. The mixture was stirred for 18 hours at 20-25 ° C. The solvent was evaporated in vacuo, the residue was dissolved in ethyl acetate, and the solution was washed successively with water, saturated aqueous sodium bicarbonate solution and then again with water. The solution was dried and concentrated in vacuo. The crude product obtained is triturated with a small amount of methanol and filtered.

The glyceryl monoester derivatives used as starting materials were prepared as follows: To a solution of the isopropylidene-protecting ester (50 mmol) in 2-methoxyethanol (300 ml) was added boric acid (500 mmol) and the mixture was heated in a water bath for 6 hours. The solvent was evaporated in vacuo and replaced with ethyl acetate. The solution was washed with water, dried and concentrated in vacuo. The ester obtained as an oily residue is purified by chromatography on a silica gel column.

K2. method: OFF. but using palmitoyl chloride instead of acetyl chloride.

K3. Method 1: A solution of 1 mmol of glyceryl monoester containing a benzylidene protecting group and 4 mmol of boric acid in 5 ml of trimethylborate was heated in a water bath for 20 minutes. The trimethylborate was evaporated and the residue was heated in a water bath for an additional 20 minutes. The resulting material was dissolved in ethyl acetate (30 mL), washed with water and evaporated. The resulting free glyceryl monoester is KI. Acylating with acetyl chloride as described in Method II.

K4. Method K3. However, the acylating agent is hexanoyl chloride.

esterification:

Ll. Method 1: To a solution of anhydrous protected amino acid (10 mmol) in -0 ° C (10 ml) in anhydrous pyridine, add 10 mmol of benzenesulfonyl chloride. The mixture is stirred for 15 minutes at 0 ° C and then 10 mmol of the appropriate alcohol is added. The reaction mixture was stirred for 18 hours at 4 ° C and the pyridine was evaporated in vacuo. The residue was dissolved in ethyl acetate (150 mL). The solution was washed successively with water, 1N aqueous hydrochloric acid, water, 2N aqueous potassium bicarbonate solution, and finally with water, dried and evaporated. The crude product is obtained.

L2. Method 1: To a stirred solution of the protected amino acid (5 mmol) in pyridine (5 mL) at 0 ° C was added phenol (10 mmol) and N, N'-dicyclohexylcarbodiimide (10 mmol) and stirred at 4 ° C for 18 h. The reaction mixture was filtered and the pyridine was evaporated in vacuo. The residue was dissolved in ethyl acetate (100 mL) and the solution was washed successively with water, 2N aqueous potassium bicarbonate solution and water again. The solution was dried and evaporated. The crude product is obtained.

L3. Method 1: To a 0 ° C solution of 10 mmol of amino acid in acetonitrile was added 10 mmol of alcohol and mmol of pyridine. 11 mmol of 11, Ν'-dicyclohexylcarbodiimide are added. The reaction mixture was stirred at 4 ° C for 18 hours, filtered, and concentrated in vacuo. The residue was taken up in EtOAc ⁵ TATB. The solution was washed successively with aqueous citric acid solution, aqueous potassium bicarbonate solution and finally water, dried and evaporated. The crude product is obtained.

L4. Method L3. A procedure according to method ¹⁰ but using acetone instead of acetonitrile as the reaction medium.

Ammonia reactions:

Ml. Method 2: The appropriate peptide methyl ester is dissolved in a minimal amount of dimethylformamide, an excess of concentrated ethanolic ammonia solution is added and the mixture is kept at 20-25 ° C for 3 days. The product is precipitated with water. 20

Hydrazine reactions:

NI. Method 10: 10 mmol of the peptide methyl ester is dissolved in 25 ml of dimethylformamide and 25 50 mmol of a 60% aqueous hydrazine hydrate solution is added. The mixture is stirred for 18 hours at 20-25 ° C, optionally concentrated in vacuo to half its original volume and the product precipitated with water.

Other reactions:

Example Method: 729 mg (1 mmol) of Boc-Tyr (Bu ¹ ) 35 - D - Ala - Gly - Phe - Leu - OH (starting material 161), 0.55 ml (7 mmol) pure, anhydrous pyridine and 0.9 ml (6.7 mmol) of freshly distilled acetic anhydride were stirred at 20-22 ° C for 10 minutes. The resulting solution was heated at 90-92 ° C for 6 hours, then cooled and mixed with 15 ml of water. The precipitated gummy material was discarded from the liquid phase and washed with water (5 x 15 mL) during decantation. The gum was then dissolved in 50 mL of ethyl acetate and the solution was successively quenched with 4 x 10 mL of 45% aqueous citric acid solution, once with 5 mL of water, 3 x 10 mL of 10% aqueous potassium bicarbonate, and 3 x 10 mL. wash with water. The solution was dried over anhydrous magnesium sulfate and evaporated. 530 mg of a solid residue 50 are obtained

Boc-Tyr (Bu ') - D-Ala-Gly-Phe-NH-CH-COCH ₃

I

Mixture of L, D, L, L and its isomers of peptide Ac. R, 0.73 (D. (system J.), 0.21 (system P.), times).

Ql. Method 555: Z.2-Tyr (Bu ') 60 -OH-dicyclohexylamine salt (55.2 g, 100 mmol) is liberated.

The free acid was dissolved in anhydrous tetrahydrofuran (300 mL) and methyl iodide (50 mL, 800 mmol) was added. 8.6 g of a 80% w / w dispersion of sodium hydride in mineral oil (i.e., 65 300 mmol of sodium hydride) are added and the mixture is stirred for 18 hours.

CHj-CHMej L, D, L, D system), 0.64 0.47 (Q. rend7

Reflux in an IS bath. Excess sodium hydride was quenched with ethyl acetate under cooling and water was added to the suspension. An almost clear solution was added. The solvent was evaporated in vacuo and the resulting aqueous residue was diluted with water (150 mL). The solution is washed twice with ether and then acidified to pH 3 with citric acid to remove inert ester impurities. The acidic mixture was extracted with ethyl acetate (400 mL then 2 x 200 mL). The extracts were combined, washed successively with water (100 mL), 10% aqueous sodium thiosulfate (100 mL), water, neutralized, and dried over anhydrous magnesium sulfate. The resulting oily residue _{15 is} suspended in 200 ml of petroleum ether at 30 ° C (boiling range: 60-80 ° C) and ethyl acetate is added to the suspension. The resulting clear solution was cooled to 4 ° C. Z-Mefyr- (Bu ') -OH is precipitated from the solution as a solid, m.p. 96-99 ° C. ²⁰

This procedure is based on the methylation method of McDermott and Benoiton [Can. J. Chem., 51, 1915 (1973)].

R j. method: 5.33 g (17 mmol) of Phe-Azleu-

_NH2 · a solution of triethylamine in 50 ml of chloroform, ²⁵ HC1 and 2.45 ml (17 mmol) of Z-NHNH-CO-Cl (3.87 g, 17 mmol). After 16 hours at room temperature, ethyl acetate (400 mL) was added. The solution was washed successively with water and 20% w aqueous citric acid solution, ³⁰ dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was chromatographed on a silica gel column eluting with 2% methanol in chloroform followed by P and Q systems. Z-Azgly- ^3: -Phe-Azleu-NH ₂ was obtained as a solid, m.p. 116-120 ° C (dec).

SKI. Procedure: A solution of 17.5 g (30 mmol) of Z-Tyr (Bzl) -OCp and 4.38 g (30 mmol) of Boc-NMe-NH _{2 in} 50 ml of dimethylformamide was kept at room temperature for 18 hours. The mixture was diluted with 500 ml of ethyl acetate, then, successively, and then washed by water, 20% w aqueous citric acid solution again. the extract was dried, concentrated in vacuo and the solid residue was treated with hydrochloric acid ethyl acetate solution of ⁴ⁱ 100 mmol. the mixture was kept at room temperature for 2 hours and the solvent evaporated under vacuum The product was a Z-Tyr (Bzl) -NHNHMe · HCl, mp .: 223-224C, _Rf. 0.82 (system D), 0.64 (system f ^50), 0.63 (system Q).

9.4 g (20 mmol) of the hydrochloride obtained above are dissolved in 200 ml of chloroform and 2.8 ml (20 mmol) of triethylamine are added, followed by 2.3 g of 55 (20 mmol) of methyl isocyanatoacetate. After stirring for 18 hours at room temperature, the solvent was evaporated in vacuo. The residue was dissolved in ethyl acetate. The solution was washed successively with water, 20% w / v aqueous citric acid solution, saturated aqueous sodium bicarbonate solution (60%), and then again with water, dried and concentrated in vacuo. The product was crystallized from a mixture of methanol and ether. M.p. 107-108 DEG C. Z-Tyr (Bzl) -Azala--Gly-OMe is obtained. 65

The structure and identification constants of starting materials prepared by the methods described above are shown in Figures 1-16. table. The methods that lead to each compound are also listed in the tables and additional information specific to the reaction conditions and the resulting material is provided in the "Note" section.

Notes 1 to 16 table:

1. Purify by precipitation.

Mp 160-161 ° C (dec).

3. Recrystallized from light petroleum (b.p. 60-80 ° C).

4. Recrystallized from a mixture of isopropanol and ether, m.p. 181-183 ° C.

5. Recrystallized from isopropanol, m.p. 193-194 ° C.

6. Chromatograph the compound on a silica gel column using chloroform and 2% methanol in chloroform as eluent.

7. The compound was chromatographed on a silica gel column using chloroform as eluent.

8. Chromatograph the product on a silica gel column eluting with chloroform and system P.

9. Recrystallized from ethyl acetate, m.p. 128-130 ° C.

10. The compound was chromatographed on a silica gel column using System G as eluent.

12. Recrystallized from a 1: 1 mixture of ethyl acetate and petroleum ether (boiling range 60-80 ° C), m.p. 96-99 ° C.

13. Recrystallized from a 1: 1 mixture of ethyl acetate and petroleum ether (boiling range 60-80 ° C), m.p. 202-203 ° C.

14. Recrystallized from ethyl acetate, m.p. 122-123 ° C.

15. The compound was chromatographed on a silica gel column using P and Q as eluent.

16. Recrystallized from ethyl acetate, m.p. 177-179 ° C.

17. Recrystallized from a 50:50 (v / v) mixture of ethyl acetate and petroleum ether (b.p. 60-80 ° C), mp 122-123 ° C.

18. Recrystallized from a 50:50 (v / v) mixture of ethyl acetate and petroleum ether (boiling range 60-80 ° C), mp 108.5-110 ° C.

20. Recrystallized from methanol, m.p. 219-220 ° C.

21. Recrystallized from a 50:50 (v / v) mixture of ethyl acetate and petroleum ether (b.p. 60-80 ° C), mp 121-122 ° C.

22. Recrystallized from a 50:50 (v / v) mixture of toluene and benzene.

23. Recrystallized from ethyl acetate / petroleum ether (50:50 v / v) 133-135 ° C.

24. The compound was chromatographed on a silica gel column using P. system as eluent.

25. The compound was chromatographed on a silica gel column eluting with 3% methanol in chloroform.

26. The compound was chromatographed on silica gel eluting with Q system.

27. The compound was recrystallized from a 50:50 (v / v) mixture of ethyl acetate and petroleum ether (boiling range 60-80 ° C), mp 122-124 ° C.

28. Recrystallized from a 50:50 (v / v) mixture of ethyl acetate and petroleum ether (b.p. 60-80 ° C), mp 115-118 ° C.

29. Recrystallized from aqueous methanol (1: 3 by volume), m.p. 162-163 ° C.

30. Recrystallized from a 50:50 (v / v) mixture of ethyl acetate and petroleum ether (boiling range 60-80 ° C), m.p. 130-132 ° C.

31. Recrystallized from a 50:50 (v / v) mixture of ethyl acetate and petroleum ether (b.p. 60-80 ° C), m.p. 156-157 ° C.

32. Freeze-dried material in the presence of hydrochloric acid.

33. Freeze-dried material.

34. The compound was chromatographed on a silica gel column using P, Q and K as eluent.

35. Recrystallized from a 50:50 (v / v) mixture of ethyl acetate and cyclohexane, m.p. 163-164 ° C.

36. Recrystallized from 50:50 methanol: ether, m.p. 130-131 ° C.

37. The ester is hydrolyzed during processing (see starting material 137).

38. The compound was chromatographed on a silica gel column using system K as eluent.

39. Recrystallized from a 50:50 (v / v) mixture of ethyl acetate and petroleum ether (b.p. 60-80 ° C), m.p. 172-174 ° C.

40. Ethyl acetate and petroleum ether 50; Recrystallized from 50% v / v, 140-141 ° C.

41. The compound was chromatographed on a silica gel column eluting with ethyl acetate-methanol. <

42. Recrystallized from ethyl acetate / petroleum ether (b.p. 60-80 ° C) 50:50, m.p. 162-165 ° C (dec).

43. Recrystallized from a 50:50 (v / v) mixture of ethyl acetate and petroleum ether (b.p. 60-80 ° C), m.p. 111-113 ° C.

Table 1

Starting material Number of reagent (s) ^used Rf · 10 ² Note number formula or formula D F Η K Q

First H-Gln-NHNH-Boc · TosOH H6 Z-Gln-NHNH-Boc 1.2 Second H-Leu-OCH ₂ CH ₂ OH · HCl E7 26th 60 30 1 Third H — Leu — OCH ₂ OCH ₂ NH ₂ · HCl away 27th 58 60 54 1 4th H-Leu-0CH ₂ CH ₂ NMeZ · HCl E7 28th 1 5th H-Leu-OCH ₂ CHCH ₂ · HCl away 29th 67 60 1 6th H-Leu-O (CH ₂ ) ₂ CHMe ₂ · HCl H4 15th 3 7th Le-Leu-OCH (CH ₂ OAc) ₂ · HCl H4 17th 58 53 31 76 8th H-Leu-OCH (CH ₂ OCOC ₅ Hi 1) 2 'HCl H4 18th 71 69 72 74 1 9th H-Leu-OCH ₂ -CHOAc -HCl 1 H4 21st 55 59 49 91 10th 1 CH ₂ OAc H-Leu-OCH ₂ -CHOCOC, ₅ H ₃₁ · HCl 1 H8 22nd 74 84 75 1 1 CH ₂ OCOC ₁₅ H ₃₁ 11th H-Leu-OPh · HCl E7 30th 1 12th H-Leu-NH- (cyclohexyl) · HQ hl 23rd 61 61 51 1 13th H — Leu- (pyrrolidine) · HCl hl 24th 4 14th H-Leu (Morpholino) · HCl hl 25th 5

Table 2

number Formula of starting material [ethod Number or formula of reagent (s) used Rf · 10 ² Note F H 15th Z-Leu-0 (CH ₂ ) ₂ CHMe ₂ L4 Z-Leu-OH + H 0 (CH ₂ ) ₂ CHMe ₂ 74 60 16th Z-Leu-CH ₂ O- (O) CH ₂ CH ₂ O CHPh L3 Z-Leu-OH + CH ₂ (MHO) ^CH 2 ^CH 2 ^O CL 1 Ph 67 70 6 17th Z-Leu-OCH (CH ₂ OAc) ₂ K3 16th 78 7 18th Z-Leu-OCH (CH ₂ OCOC ₅ i) ₂ K4 16th 7 19th Z-Leu-OCH ₂ -CHO \ 1 / CMe ₂ CH ₂ 0 L4 Z-Leu-OH + H0CH ₂ -CH0x 1 .CMe ₂ ch ₂ o ^z 65 68 67 20th Z-Leu-OCH ₂ CHOHCH ₂ OH WHO 19th 8 21st Z-Leu-OCH3 -CHOAc I WHO 20th 75 1 CH ₂ OAc 22nd Z-Leu-OCH ₂ -CHOCOCt _S H ₃₁ | K2 20th 81 70 1 CH ₂ OCOC ₁₅ H ₃ i 23rd Z-Leu -NH- (cyclohexyl) B4 Z-Leu-OH + cyclohexylamine 79 73 9 24th Z-Leu- (pyrrolidino) B4 Z-Leu-OH + pyrrolidine 73 71 25th Z-Leu- (morpholino) B4 Z-Leu-OH + morpholine 79 59

Table 3

number Starting material Method Reagents used R _f · 10 ² Note Q formula formula D F H

26th Boc-Leu-O (CH ₂ ) ₂ 0H L1 Boc-Leu-OH + HO (CH ₂ ) ₂ OH 27th Boc-Leu- (OCH ₂ ) ₂ NH ₂ L1 Boc-Leu-OH + H 0 (CH ₂ ) ₂ NH ₂ 73 74 70 10 28th Boc-Leu-O (CH ₂ ) ₂ NMeZ L1 Boc-Leu-OH + HO (CH ₂ ) ₂ NMeZ 7 29th Boc-Leu-OCH ₂ -CHCH ₂ L1 Boc-Leu-OH + H0CH ₂ -CHCH ₂ 11 30th Boc-Leu-OPh L2 Boc-Leu-OH + phenol 10

44. The compound was chromatographed on a silica gel column eluting with chloroform containing 1% methanol. 121-122 ° C. 60

45. The compound was chromatographed on a silica gel column using Q and K as eluent.

46. Recrystallized from ethyl acetate, m.p.

248-250 ° C. 65

47. The compound was purified on Sephadex LH20 gel using dimethylformamide as solvent.

48. Recrystallized from a 50:50 (v / v) mixture of ethyl acetate and petroleum ether (boiling range 60-80 ° C), m.p. 152-155 ° C.

49. Recrystallized from a 50:50 (v / v) mixture of ethyl acetate and petroleum ether (boiling range 60-80 ° C), m.p. 164-168 ° C.

50. The compound was chromatographed on a silica gel column eluting with 5% methanol in ethyl acetate.

As in Figures 1-16. Table 6 shows the columns headed "Number or formula of reagent (s) used", starting materials are described starting with the corresponding individual amino acids or peptide fragments. It can also be seen from the tables that many starting materials serve as reagents for the preparation of further starting materials.

Table 4

Starting material The formula number of the reagent (s) used is the number or formula D F Η K Q

31st ZMeTyrCBuŰ-OH Q1 Z-Tyr ( ¹ ) - OH (MeI / NaH) DMF 62 46 27 12 32nd H — Azphe — Leu — OMe · HCl away Boc-Azphe-Leu-OMe; 77 72 1 33rd H-Gln-Gln-NHNH-Boc Hll 34th 23 50 1 34th Z-Gln-NHNH-Boc A4 Z-Gln-OCp + 1. 87 49 20 1 35th H-Phe-Azleu-NH ₂ F2 38th 1 36th H-Phe-Nle-OMe · TosOH H6 37th 63 59 65 1 37th Z-Phe-Nle-OMe B5 Z-Phe-OH + H-Nle- -OMe · HCl 88 77 78 1 38th Boc-Phe-Azleu-NH ₂ D2 Boc-Phe-OH + H-Azleu-NH ₂ * HCl 13 39th H — Pro — Lys (Z) —OM · HC1 E7 40th 67 30 1 40th Boc-Pro-Lys (Z) -OMe B1 Boc-Pro-OH + H-Lys (Z) -OMe · HCl 84 60 77 41st H-D-Ala-Gly-Phe-OMe hl 42nd 35 22 42nd Z-D-Ala-Gly-Phe-OMe THE 3 Z-D-Ala-OCp + H-Gly-Phe-OMe · HCl 78 62 64 58 14 43rd Z-Arg (Z ₂ ) (- Pro-Lys (Z) -OH) J3 44th 64 20 20 1 44th Z-Arg (Z ₂ ) —Pro — Lys (Z) —OMe A1 Z-Arg (Z ₂ ) ONSu + 39. 85 78 80 7 45th H-Azgly-Phe-Azleu-NH ₂ · HCl H5 46th 33 23 92 1 46th Z-Azgly-Phe-Azleu-NH ₂ R j Z — Azgly — Cl + 35. 72 73 94 22 15 47th H-Gly-Azphe-Leu-OMe · HCl E4 50th 56 36 90 19 1 48th G-Gly — Phe — Nle-OMe · TosOH H6 49th 1 49th Z-Gly-Phe-Nle-OMe B5 Z-Gly-OH + 36. 81 79 71 1 50th Boc-Gly-Azphe-Leu-OMe; D2 Boc — Gly — OH + 32. 77 76 16 51st Boc-Gly-Gly-Gly-OH H2 52nd 39 45 1

Continuation of Table 4

μ .......- -.....—. ------------ --------- The formula of the starting material number Reagent (s) used Number or formula of the method D F Rf H 10 ² K Megd quotes 51 A. Boc-Gly-Gly-Gly-ONa J4 51B. 35 41 1 51B. Boc-Gly-Gly-Gly-OMe B2 Boc-Gly-OH + H- (Gly) ₂ -OMe · HCl 60 57 52. Boc-Gly -Gly-Gly --OBzl A1 Boc-Gly-OCp + H- (Gly) ₂ -OBzl · HCl 68 63 39 53. H-Gly-Phe-Leu-OMe Coll. Czech. Chem. Comm. 29, 2633 (1964) 33 32 74 29 18 54. Z-Pro-Gin-NHNH-Boc B5 Z-Pro-OH + 33. 67 93 20 55. · H-D-SerCBu ^ -Gly Phe-OMe hl 57th 56 44 95 20 1 56. H-D-Se ^ Bu ^ -Gly- -Phe (6H) -OMe G1 57th 64 47 45 1 57. Z-D-SerCBu ^ -Gly Phe-OMe D2 Z-D-Ser (Bu ') - OH + H-Gly- -Phe-OMe · HCl 73 62 63 40 21 58. Z-Tyr (Bzl) —Azala Gly-OMe SKI Z-Tyr (Bzl) NHNHMe + + OCNCH ₂ CO ₂ Me 77 72 80 59 1 59. Z — Tyr (Bzl) —Azala— —Gly — NHNH ₂ NI 58th 68 51 45 27

Table 5 number

Formula of starting material

Method ^The reagents used) or the number of the formula D

R _f ♦ 10 ²

Η Q.

Leave a ·

Q

60th H-D-Ala-Gly-Phe-Leu-OMe · HC1 H10 61st 88 1 61st Z — D — Ala — Gly —Phe —Leu — OMe A4 Z-D-Ala-OCp + 53. 73 62 48 1 62nd H-D-Asp-Gly-Phe-Leu-OMe; H2 63. 51 67 1 63rd Z-D-Asp (OBzl) -Gly-Phe-Leu-Ome A4 Z — D — Asp (OBzl) —OCp + 53. 22 64th H-D-Leu-Gly-Phe-Leu-OMe · TFA F2 65th 1 65th Boc-D-Leu-Gly-Phe-Leu-OMe; B4 Boc-D-Leu-OH + 53. 78 79 74 15

Continuation of Table 5

Starting material ,,,, Method number formula Number or formula of reagent (s) used Rf · 10 ² Note D H K the 66th H-D-Lys (Boc) -Gly-Phe-Leu-OMe; H2 67th 1 67th Z-D-Lys (Boc) -Gly-Phe-Leu-OMe; B4 Z — D — Lys (Boc) —OH + 53. 76 76 65 68th H-D-Met-Gly-Phe-Leu-OMe · HCl E5 69th 69th Boc-D-Met-Gly-Phe-Leu-OMe; A1 Boc-D-Met-OCp + 53. 70th H — D —SerCBu ¹ ) —Gly — Phe — Leu-OMe H3 71st 61 57 88 46 1 71st Z-D-Ser / Bu *) - Gly-Phe-Leu-OMe; B2 Z-D-SerCBu ^ -OH + 53. 79 75 74 23 72nd H-D-Thr-Gly-Phe-Leu-OMe; H2 73rd 58 36 71 20 1 73rd Z-D-Thr-Gly-Phe-Leu-OMe; cl Z-D-Thr-NHNH ₂ + 53. 70 63 10 74th H-D-Trp-Gly-Phe-Leu-OMe · TFA F3 75th 61 55 27 1 75th Boc-D-Tip-Gly-Phe-Leu-OMe; THE 2 Boc-D-Trp-OCp + 53. 79 70 94 60 24

Table 6

The formula of the starting material number Reagent (s) used R _f · 10 ² Comment samma or formula D F H Q 76th HD-Ala-Azgly-Phe-Azleu-NH ₂ · HCl H5 79th 34 22 1 77th H-D-Ala-Gly-Phe -Nle-OMe · TosOH H6 80th 78th H-D-Ala-Gly-Phe (6H) -Leu-OMe G1 60th 79th Z-D-Ala-Azgly-Phe-Azleu-NH ₂ D2 Z-D-Alá-OH + 45. 64 69 31 80th Z-D-Ala-Gly-Phe-Nle-OMe A5 Z-D-Alá-OCp + 48. 76 77 56 1 81st H-0-Al-Gly-Phe-NLe-OMe · HCl E7 82nd 60 1 82nd Boc-0-Ala-Gly-Phe-Nle-OMe A5 Boc— / 3 — Ala — OCp + 48. 69 69 49 25 83rd Z-Me-Tyr / D SerCBu Bu ^ ^ Gly-Phe (6H) OH J3 84th 68 67 50 26 84th Z — Me — Tyr / Bu ¹ ) —D —Safe ¹ ) —Gly— —Phe (6H) OMe B1 31st + 56th 78 77 70 85th H-D-SerfBu ¹ ) -Gly-Azphe-Leu -OMe · HCl H5 86th 62 57 91 86th ZD — Ser / Bu ¹ ) —Gly-Azphe —Leu-OMe D2 Z — D — SeifBu ¹ ) —OH + 47. 80 76 4 ' 24

Table 7

The formula of the starting material number .. ,, For the reagent used) Method. . , 7 samba or image D F Rf H 10 ² K Q Give one. ing 87. Z — Tyr — D — Ser — Gly — Phe — OH F2 82nd 56 52 1 88. Z-TyriBu 1 -D-Seribu ¹ ) -Gly-Phe-OH J2 89th 63 53 62 85 1 89. Z-TyrCBu ^ -D-Se ^ Bu ¹ ) —Gly — Phe — OMe A1 Z-TyrCBu ^ -OCp + 55. 26 90. Boc-Tyr-D-Ala-Gly-Phe-OH hl 93rd 62 54 55 71 16 27 91. Boc-Tyr-D-Ser / Bu ¹ ) - -Gly-Phe-OH hl 95th 63 59 55 76 13 ' 1 92. Boc-Tyr-D-SerCBu ¹ ) -Gly- -Phe- (tetrahydropyrazole) · HCl H5 97th 60 41 1 93. Boc-Tyr (Bzl) -D-Ala-Gly-Phe-OH J3 94th 58 50 32 28 94. Boc-Tyr (Bzl) -D-Ala -Gly-Phe-OMe D2 Boc — Tyr (Bzl) —OH + 41. 78 70 29 95. Boc-Tyr (Bzl) -D-Ser (Bu ¹ ) -Gly-Phe-OH J3 96th 61 60 58 72 11 30 96. Boc-Tyr (Bzl) -D-SeríBu ¹ ) - -Gly-Phe-OMe D2 Boc — Tyr (Bzl) —OH + 55. 77 67 65 73 31 97. Boc-Tyr (Bzl) ~ D-Ser (Bu ') ~ -Gly-Phe- (N-Z-tetrahydropyrazole) D2 95. + N-Z-tetrahydropyrazole · HCl 84 79 60 25 98. Boc-TyrfBu ^ -D-Ala-Gly-Phe-OH J3 99th 10 55 48 1 99. Boc — Tyr (Bu ^l ) ~ D — Al— —Gly-Phe-OMe B4 Boc-Tyr (Bu ') - OH + 41. 74 98 57 1 100. Boc — TyrCBu ¹ ) —D — Seribu ¹ ) - —Gly — Phe (6H) OH J3 One hundred and first 68 67 27 1 101. Boc-TyrCBu ^ -D-SerCBu ¹ ) - —Gly — Phe (6H) -OMe B1 Boc-TyrCBu ^ -OH + 56. 78 77 Ί 25

Table 8

The formula of the starting material number

Method Number or formula of reagent (s) used D

R _f · 10 ²

Η Q.

102. H-Me-Tyr-D-Ser-Gly -Phe (6Hj-Leu-0Me

104th

43

Comment

Continuation of Table 8

Starting material Method reagent (s) used number or formula D R _f - 10 ² Comment number formula H K 103rd H-Me-D-TyrCBu ^ Se ^ Bu *) - Gly-Phe-Leu-OMe; H3 107th 1 104th H-Me-Tyr (Bu ^t ) -D-Ser (Bu ^t ) -Gly-Phe (6H) -Leu-OMe H2 108th 66 63 50 1 105th Z — Me — Tyr — D — Al — Gly — Phe — Leu — OMe F2 106th 64 62 94 51 1 106th Z-Me-TyrfBu ¹ ) -D-Ala-Gly -Phe Leu -OMe B1 Z-Mc-Tyr (Bu ') - OH + 60. 50 76 95 67 1 107th Z-Me-Tyr (Bu ^t ) —D — Ser (Bu ^t ) - —Gly — Phe (6H) —Leu-OMe B1 83 + H-Leu-OMe · HCl 87 77 75 25 108th Z-Me-TyKBu ^ -D-SeríBu ¹ ) —Gly — Phe — Leu — OMe D3 Z-Me-Tyr (Bu ^l ) -ONSu + 70. 80 77 82 1 109th H-Tyr-D-Lys (Boc) -Gly-Phe -Leu-OMe · HC1 H9 129th 60 49 14 1.32 110th H - Tyr - D - Ser - Gly - Phe (6H) - - Leu - OMe F2 177th 65 1 111th H-Tyr-D-Ser (Bu ^t ) -Gly-Phe- -Leu-OMe hl 141st 66 97 25 33 112th H-Tyr (Bzl) -D-Ala-Gly-Phe -Leu-OMe · HC1 away 152nd 66 54 98 31 1 Table 9 Starting material 0 tg Rf 10 ² Comment number i § O A B C D F H K Q

113th H-Tyr (Bzl) -D-Ser-Gly-Phe-Leu -OCH ₂ CH ₂ OH · TFA F2 154th 65 68 66 60 51 52 90 1 114th H-Tyr (Bzl) -D-Ser-Gly-Phe-Leu -OCH ₂ CH ₂ OAc · TFA F2 155th 78 88 77 62 52 62 90 1 115th H-Tyr (Bzl) -D-Ser-Gly-Phe-Leu -OCH ₂ CH ₂ OCOCt _s H ₃ 1 · TFA F2 156th 73 79 72 68 56 59 95 1 116th H-lyr (Bzl) -D-Ser-Gly-Phe-Leu -OCH (CH ₂ OAc) ₂ · TFA RIP 157th 62 68 61 63 54 50 83 1 117th H-Tyr (Bzl) -D-Ser-Gly-Phe-Leu -OCH (CH ₂ OCOC _s Hn) ₂ · HCl RIP 158th 59 67 60 66 52 1 118th H-Tyr (Bzl) -D-Ser-Gly-Phe-Leu -OCH ₂ -CHOAc · TFA I F2 159th 64 60 54 56 SS 46 86 1 1 CH ₂ OA _c 119th H-iyr (Bzl) -D-Ser-Gly-Phe-Leu -OCH ₂ -CHOCOCl ₅ H ₃ i -TFA F2 160th 66 68 58 56 55 58 90

CH ₂ OCOC ₁₅ H ₃ i

Table 10

Starting material number formula Módszc _{; r} Number or formula of reagent (s) used _; D R _f · 10 ² j Note H K Q 120. H-Tyr (Bu ^t ) -D - Ser (Bu ^t ) -Gly -PheLeuO (CH ₂ ) ₂ CHMe ₂ hl 142nd 58 9B 49 1 121. H-TyrCBu ⁴ ) -D-SerfBu ¹ ) -Gly-Phe-Leu (tetrahydropyrrolo) hl 143rd 60 98 28 1 1-22. H-TyriBu ^ -D-Se ^ Bu ¹ ) —Gly — Phe — Pro — NHEt hl 144th 57 84 29 34 123. Ac-Tyr-D-Ala-Gly-Phe-Leu-OMe H12 125th 63 98 124. Z-Tyr-D-Ser-Gly-Phe-Leu-OBu * D2 87. + H-Leu-OBu * 66 86 40 15 125. Ac-Tyr (Bzl) -D-Ala-Gly -Phe-Leu-OMe A4 Ac-OCp + 112. 72 98 1 126. Z-Tyr (Bzl) -D-Ala-Gly -Phe-Leu-OMe A1 Z — Tyr (Bzl) —OCp + 60. 85 81 72 1 127. Z — Tyr (Bzl) —D — Asp-Gly— —Phe — Leu — Ome A1 Z — Tyr (Bzl) —OCp + 62. 28 24 128. Z — Tyr (Bzl) —Azala — Gly— —Phe — Leu-OMe cl 59. + H — Phe — Leu — OMe · HCl 78 61 35 129. Z-Tyr (Bd) -D-Lys (Boc) - Gly-Phe-Leu-OMe; ' THE 2 Z-Tyr (Bzl) —OCp + 66. 72 130. Z — Tyr (Bzl) —D — Thr — Gly— —Phe — Leu-OMe THE 2 Z — Tyr (Bzl) —OCp + 72. 81 75 70 131. Z-Tyr (Bzl) -D-Trp-Gly-Phe-Leu-OMe A4 Z — Tyr (Bzl) —OCp + 74. 82 71 70 1 132. Z-Tyr (Bzl) -D-Ala-Azgly -Phe-Azleu-NH ₂ A1 Z-Tyr (Bzl) - OCp + 76. 69 60 23 36 133. Z — Tyr (Bd) —D — Ala — Gly— —Phe — Leu — NH ₂ ml 126th 79 75 75 1 134. Z-Tyr (Bzl) -D-Ala-Gly -Phe-Nle-OMe A4 Z-Tyr (Bzl) -OCp + 77. 70 1 135. Z-Tyr (Bzl) -0-Ala-Gly -Phe Nle-OMe A4 Z — Tyr (Bzl) —OCp + 81. 87 72 68 1 136. Z — Tyr (Bzl) —D — Ala-Gly— -Phe (6H) -Leu-OH A1 70 50 90 37, 38 137. Z — Tyf (Bzl) —D-Ser-Gly— -Azphe-Leu-OMe; E8 139th 85 24 57 25 138. Z — Tyr (Bzl) —D — Ala-Gly— -Phe (6H) -Leu-OMe A1 Z — Tyr (Bd) —OCp + 78. 71 70 60 25

Table 10 continued number

Formula of starting material

Method Number or formula of reagents used

Rf * 10 NOTES Η K Q. **

139. Z-Tyr (Bzl) —D — Ser / Bu ¹ ) -

Azphe-Gly-Leu-OMe; A1 Z-Tyr (Bzl) -OCp + 85. 78 79 77 140th Z-Tyr (Z) -D-Leu-Gly -Phe-Leu-OMe; A4 Z — Tyr (Z) —ONp + 64. 80 76 95 73 24 141st Z-Tyr (Z) -D-SerCBu ¹ ) - -Gly -Phe-Leu-OMe THE 2 Z — Tyr (Z) —ONp + 70. 76 98 75 142nd Z-Tyr (Bu ^t ) ^ D-Ser (But) - —Gly — Phe — Leu— —O (CH ₂ ) ₂ —CHMe ₂ D2 88. + 6. 73 98 72 1 143rd Z-Tyr (But) -D-Ser (Bu *) - Gly-Phe-Leu (tetrahydro) D2 88th + 13th. 72 66 1 144th Z-Tyr (Bu ') - D-SerCBu ¹ ) -Gly-Phe-Pro-NHEt D2 88. + H — Pro — NHEt · HCl 68 98 67 1 145th Boc-Tyr-D -Alá-Gly-Phe -Azleu-NH ₂ D2 90. + H — Azleu — NH ₂ · HQ 74 73 86 39 146th Boc-Tyr-D-Ala-Gly-Phe -DL-NH- (2-oxo-tetrahydrofuran-3-yl) dL 90. + DL-H ₂ N- (2-oxo-tetrahydrofuran-3-yl) · HBr 41 1 147th Boc-Tyr-D-Ala-Gly -Phe-Met-OMe D2 90. + H-Met-OMe · HCl 70 76 36 148th Boc-Tyr-D-Met-Gly-Phe-Leu-OMe A4 Boc-Tyr-OCp + 68. 76 71 55 41 149th Boc-Tyr-D-Ser (Bu ¹ ) -Gly-Phe-Azleu-NH ₂ D2 91. + H — Azleu — NH ₂ · HCl 75 80 86 15 42 150th Boc-Tyr-D-SerfBu *) - -Gly-Phe-Azpro-NHEt H2 153rd 77 63 1 151st Boc-Tyr-D-Ser (Bu *) - Gly-Phe-Met-OMe D2 91. + H-Met-OMe · HCl 74 81 53 43 152nd Boc-Tyr (Bzl) -D-Ala -Gly-Phe-Leu-OMe bl Boc-Tyr (Bzl) -OH + 60. 81 64 97 65 153rd Boc-TyrfBzlj-D-SerCBu ⁴ ) —Gly — Phe — Azpro — NHEt D2 95. + H-Azpro-NHEt 63 56 53 25

Table 11

Starting material Formula: Number Boc-Tyr (Bzl) -D-Seribu ¹ ) - -Gly-Phe-Leu-X, where X: Method Number of reagent (s) used R _f · 10 ² ΕΛ 'EB D F H Q 154. -O (CH ₂ ) ₂ -OH B3 95. + 2. 86 54 73 58 24 155. -O (CH ₂ ) ₂ OAc WHO 154th 78 73 64 25 156. -O (CH ₂ ) ₂ -OCOCi _S H ₃₁ K2 154th 77 76 67 25 157. -OCH (CH ₂ OAc) ₂ D2 95. + 7. 69 68 72 71 1 158. -OCH (CH ₂ OCOC ₅ Hj i) ₂ D2 95th + 8th 82 75 76 72 1 159. -OCH ₂ -CHOAc D2 95. + 9. 74 73 64 44 CH ₂ OAc 160. -OCH ₂ -CHOCOC ₁₅ H ₃₁ D2 95. + 10. 88 81 44

I.

CH ₂ OCOC _1S H ₃₁

Table 12

Starting material Formula: g number Boc-TyriBu ^ -D-Ala-Gly-Phe-Leu-X, where X: Jg Number or formula of reagent (s) used D F R _f · 10 ² Η K Q Comment 161. -OH J5 162nd 55 93 1 162. -Me B2 Boc-TyrCBu ¹ ) - OH + 60. 78 76 69 7 163. -O (CH ₂ ) ₂ OH B1 98. + '2. 88 66 60 24 164. -O (CH ₂ ) ₂ NH ₂ B1 98. + 3. 74 68 56 24 165. -O (CH ₂ ) ₂ NMeZ B1 98. + 4. 24 166. -OCH ₂ -CHCH ₂ B1 98. + 5. 83 64 66 1 167. —OPh B1 98. + 11. 24, 45 168. -NH (CH ₂ ) ₂ OH B1 161. + NH ₂ (CH ₂ ) ₂ OH 72 67 48 24 169. —NH (CH ₂ ) ₂ NHMe B1 161. + NH ₂ (CH ₂ ) ₂ NHMe 10 96 46 170. -NH (CH ₂ ) ₂ NMe ₂ B1 161. + NH ₂ (CH ₂ ) ₂ NMe ₂ 23 30 99 38 171. -No ₂ B1 161. + NHEt ₂ 66 63 30 24 172. cyclohexylamino- D2 98. +12. 60 65 1 173. pyrrolidine- D2 98. +13. 72 71 75 55 1 174. morpholino D2 98. +14. 72 71 75 55 24

Table 13

Starting material Reagent (s) used Rf 10 ² remarks ,,,. . Moaszer formula of sam number or formula D H K Q ing 175th Boc- —TyrCBu ^ -D-Ala-Gly-Phe— Pl 161st 73 64 47 Ac -DL-Leu-Me • 176th Boc-Tyr (Bu *) - D-Ala-Gly-Phe -Pro-NHEt D2 98. 4 H — Pro — NHEt 65 98 45 24 177th Boc-D-SerCBu TyrCBu ^ ^ -Gly —Phe (6H) —Leu-OMe Bl 100. + H — Leu-OMe · HCl 87 77 76 44

Table 14

Starting material Formula: X-Tyr (Bzl) -Damba -Ala-Gly-Phe-Leu-OMe, where X: Method Number or formula of reagent (s) used D F R _f · H 10 ² HP N&Q K the 178. Η-β-Alá · HC1 away 179th 61 41 78 1 179. Boc-0-Ala al Boc-0-Ala-OCp +112. 82 70 98 65 1 180. Z — Asp (OBzl) —Gly — Gly — Gly A4 Z — Asp (OBzl) —OCp + 182. 62 68 36 1 181. Z-GluCOBu *) al Z-GluCOBu *) - OCp + 112. 84 75 79 28 182. H-Gly-Gly-Gly · HCl E2 184th 55 7 73 38 183. Z — Gly — Gly — Gly cl Z-Gly-Gly-Gly-NHNH ₂ + 112. 70 57 60 26 26 184. Boc — Gly — Gly — Gly B2 Boc — Gly — Gly — Gly — OH + 112. 78 55 21 1 185. H-Gly-Gly · HCl E2 186th 58 16 68 1 186. Boc — Gly — Gly B2 Boc — Gly-Gly — OH + 112. 60 61 90 38 1 187. Z — Leu — Leu — Leu bl Z-Leu-Leu-Leu-OH + 112. 71 76 96 61 47 188. Z-Lys-Gly-Gly-Gly E4 189th 61 1 189. Z — Lys (Boc) —Gly — Gly — Gly A4 Z — Lys (Boc) —OCp + 182. 64 66 56 31 1 190. Z-Lys · HCl E4 193rd 54 80 87 15 1 191. Z — Asp (OBzl) Z-Lys al Z-Asp (OBzl) -OCp +190. 85 95 64 192. Z-D-Lys (Boc) bl Z-D-Lys (BOc) -OH + 112. 80 80 97 67 1 193. Z-Lys (Boc) al Z-Lys (Boc) -OCp +112. 76 74 79 66 71 69 24 1 194. Z-Glu-Gly-OBzl Z-Lys B2 Z-Glu-Gly-OBzl + 190. 82 72 71

Continuation of Table 14

Starting material Beat Rf · 10 ² Formula: X-Tyr (Bzl) -D- (75 Reagent (s) used Another- number -Ala-Gly-Phe-LeuOMe, *SHE number or formula D F Η K Q where X: 3

195th Z-Gly-Gly-Gly Z-Lys cl Z-Gly-Gly-Gly-NHNH ₂ +190. 68 68 68 39 196th Z-Lys (Z) Z-Lys B1 Z-Lys (Z) Z-Lys-OH + 112. 82 66 98 60 197th Boc-Phe Z-Lys D2 Boc-Phe Z — Lys — OH + 112. 82 74 90 96 75

Table 15

Starting material Method Reagents used)

SZCiT - formula number or formula D F H K Q 1%. Z-Arg (Z ₂ ) -Pro-Lys (Z) -Pro- -Gin-Gln-Tyr-D-Al-Giy Phe-Leu-OMe D2 Example 43 + 38 199th Z-Arg (Z ₂ ) -Pro-Lys (Z) -Tyr-D-Ala-Gly-Phe -Len-OMe D2 43. + 112. 49 62 200th Z-Asp-Tyr-D - Ala-Gly-Phe-Leu-OMe · TFA F2 Two hundred and first 60 30 Two hundred and first AspCOBu ^ Z-Tyr-D-Ala —Gly — Phe — Leu-OMe A2 112. + Z — Asp— (OBu *) ONSu 73 70 40 202nd H-GMOBu ^ -Tyr-B-Ala-Gly-Phe-Leu-OMe Hl 181st 69 63 56 56 2®3. Z-Gly-Gly-Gly-Tyr-D-Ser- -Gly-Phe (6H) -Leu-OMe Cl 110. + Z- (Gly) ₃ -NHNH ₂ 204th loc-Gly -Gly-Gly-Gly-Gly- -Gly-Tyr-D-Ala -Gly-Phe-Leu-OMe B2 Example 51 + 69 75 205th Boc-Gly-Gly-Gly-Me-ThirlBu ¹ ) -D-SeKBuH-Gty-Phe-Leu-OMe B2 51a. +103. 70 71 44 206th H-D-Lys (Boc) -Tyr-D-Ala -Gly-Phe-Leu-OMe H3 192nd 65 46 96 18 207th H-Lys (Boc) -Tyr-D-Ala-Gly -Phe-Leu-OMe H9 193rd 68 57 55 20 208th Boc-Phe H-Lys-Tyr-D-Alá —Gly — Phe-Leu-OMe Hl 197th 65 72 60 98 25 209th Z-Lys-Me-Tyr-D-Ser-Gly-Phe-Leu-OMe F2 211th 59 65 42

Continuation of Table 15 The formula of the starting material number Method Reagent (s) used Rf · 10 Notes number or formula D F Η K QL

210. Z-Lys (Z) -Tyr-D-Ala-Gly -Phe-DL-NH- (2-oxo-

tetrahydrofuran-3-yl) D2 Example 46 + Z-Lys (Z) -OH 69 66 67 89 211th Z — Lys (Boc) —Me — Ty ^ Bu *) - D-SerCBfoj-Gly— —Phe —Leu — OMe B2 103. + Z — Lys (Boc) —OH 84 78 77 24 212th Boc-Ly s (Boc) -Tyr-D -Alá-Gly-Phe-Azleu-NH ₂ D2 Boc — Lys (Boc) —OH + 45. pl. 63 64 31 49 213th Boc-Lys (Boc) -Tyr-D-Ala -Gly-Phe-Met-OMe D2 Boc — Lys (Boc) —OH + 51. pl. 70 63 46 50 214th Boc — Lys (Boc) —Tyr — Azala— —Gly —Phe — Leu — OMe D2 Boc — Lys (Boc) —OH + 10. pl. 73 78 51 51 215th Boc-Lys (Boc) -Tyr-D-Ser- -Gly-Phe-Azleu-NH ₂ D2 Boc-Lys (Boc) -OH +55. e.g. 74 73 68 15 216th Boc-Lys (Boc) -Tyr-D-Ser- -Gly-Phe-Met-OMe D2 Boc — Lys (Boc) —OH + 58. pl. 70 60 48 25 217th Z-Pro-Gln-Gln-Tyr-D-Ala-Gly-Phe-Leu-OMe; cl Example 54 + 47 67 13 48 218th H-Tyr-D-Ala-Gly-Phe-Leu -Thr (Bzl) -OBzl · TFA F2 223rd 68 49 24 1

Table 16

Holiday material Number Formula: Boc-Tyr (Bu ') - D-Ala- Method Number or formula of reagents used D R-10 ² Note -Gly — Phe — Leu — X, where X: H Q 219th D-Ala-OMe B1 161. + H-D Ala OMe · HC1 80 74 55 25 220th Ala-OMe B1 161. + H-Ala-OMe · HCl 72 78 64 1 221st Gly-OMe B1 161. + H-Gly-OMe · HC1 87 75 61 25 222nd D-Thr-OBu * B1 161. + H-D-Thr-OBu * 82 49 25 223rd Thr (Bzl) -OBzl B1 161. + H — Thr (Bzl) —OBzl · HCl 85 66 64 44

For preparing compounds (1) of the formula ³² 1-93. example.

1-42. example

The protected polypeptide is H1-H13. method. The products listed in Table 17 are obtained.

The protecting groups are cleaved by the following methods:

Method Hl: Dissolve the starting material in ethanol containing up to 25% water and add 150 mg / g of 5% palladium on charcoal in nitrogen atmosphere and stir slightly at 20-25 ° C with stirring. a stream of hydrogen was passed through the stream. The reaction is usually complete after 5-6 hours. The hydrogen atmosphere is replaced by a nitrogen atmosphere, the catalyst is filtered off over diatomaceous earth and the filtrate is concentrated in vacuo. The desired product is obtained. 5

H2. Method III: Method H1, but max. Methanol containing 25% by volume of water was used.

H3. Method III: Method H1, but using methanol as the reaction medium. 10 H4. Method III: Method H1, but with 1 molar equivalent of hydrochloric acid and max. Ethanol containing 25% by volume of water was used.

H5. Method 15: Method H1 was used, but 15 molar equivalents of hydrochloric acid and max. Methanol containing 25% by volume of water was used.

H6. Method 1: Method H1, but using 1 molar equivalent of p-toluenesulfonic acid dimethylformamide as the reaction medium.

H7. Method 1: Method H1, except that 1 molar equivalent of hydrochloric acid was added to the reaction medium and methanol was used as the reaction medium. 25 H8. Method A: Method H1, but using a 50:50 by volume mixture of dimethylformamide and butanol as the reaction medium.

H9. Method A: Method H1 was carried out using 90 or 95% by volume of 30% acetic acid in the reaction medium.

H10. Method H9. However, 1-2 molar equivalents of hydrochloric acid are added to the reaction medium.

Hll. Method 35: Method H1 is used, but max. Dimethylformamide containing 25% by volume of water was used.

H12. Method A: Method H1 was used except that the reaction medium was a 50:50 (v / v) mixture of ethanol and chloroform (25% in water).

H13. Method A: Method H1, but using a 50:50 by volume mixture of methanol and dimethylformamide as the reaction medium.

Notes to Table 17:

1. Hydrochloride salt

2. Dihydrochloride salt

3. Trihydrochloride salt

4. Acetate salt

5. Diacetate salt

6. Triacetate salt

7. Trifluoroacetate salt

8. The mixture was filtered through diatomaceous earth and evaporated.

9. Isolate the product by ether precipitation.

10. The product is isolated by precipitation with methanol-ether.

11. The product is isolated by precipitation with ethyl acetate in ether.

12. Obtained as a freeze-dried powder (lyophilization medium: water or a mixture of water and tert-butanol).

13. See note 12, in the presence of hydrochloric acid.

14. See Note 12 in the presence of acetic acid.

15. Purify by chromatography on silica gel (solvent: 25% v / v methanol-chloroform).

16. See note 15, using solvent K as solvent.

17. Purified on Sephadex G15 gel (solvent: 5% v / v aqueous acetic acid).

18. See Note 17, Sephadex G25 Gel is used.

19. Electrophoresis at pH 6.5 displaces 0.78 units against the histidine standard.

Melting point 143 ° C.

Melting point: 157 ° C.

Mp 223 ° C (dec).

23. M.p .: 81-83 ° C.

M.p. 87 ° C (dec).

M.p .: 146-148 ° C.

26. Mp 98 ° C (dec).

27. M.p .: 168 ° C.

28. M.p .: 218-219 ° C.

Table 17

examples RfXlO ² Kiin. Mode- · you da Product material agent ticket- number A B C D F Η K Q number ing

1. H-Met-Tyr-D-Ala-Gly-

-Phe-Leu-OMe; 50 41 87 16 105. H7. 1.16 Second H-Tyr-D-Ala-Azgly -Phe-Azleu-NH ₂ 32 55 55 20 60 132nd H5. 1, 10 Third H-Tyr-D-Ala-Gly-Phe-Leu-NH CXCHj, 65 73. pl. H9. 5.12 4th H-Tyr-D-Ala-Gly-Phe-Leu-O (CH ₂ ) ₁ NHMe 30 69 77 74. pl. H9. 2.13

Continuation of Table 17

Example number product the RfXlO ² Kiin. material number Mode- agent Comment THE B C D F H K the 5th H - Tyr - D - Ala - Gly - Phe - Leu - NH ₂ 65 10 133rd H9. 16 6th H-Tyr-D-Ala-Gly-Phe -Nle-OMe 61 30 83 134th H9. 4 7th H-Tyr-D-Ala-Gly-Phe (6H) -Leu-OH 60 22 74 136th H9. 12 8th H-Tyr-D-Ala-Gly- -Phe (6H) -Leu-OMe 61 48 137th H2. 8 9th H - Tyr - D - Asp - Gly - Phe - Leu - OMe 57 90 127th H2. 12, 20 10th H — Tyr — Azala — Gly— —Phe — Leu-OMe 66 52 85 128th H5. 1/8 11th H-Tyr-D-Leu-Gly-Phe-Leu-OMe; 60 59 50 84 17 140th H9. 1, 13 12th H - Tyr - D - Ser - Gly - Azzhe - Leu - OMe 71 79 77 66 53 86 138th H5. 1, 9, 21 13th H - Tyr - D - Ser - Gly - Ph - Leu - O (CH ₂ ) ₂ OH 67 67 50 60 50 46 95 113th Hll. 7, 11, 22 14th H - Tyr - D - Ser - Gly - Ph - Leu - O (CH ₂ OAc 67 71 65 67 50 87 114th Hll. 7, 8, 23 15th H-Tyr-D-Ser-Gly-Phe-Leu-OCCH ₂ ) ₂ OC (O) C ₁₅ H ₃₁ 59 68 62 59 49 44 89 115th Hll. 7, 8, 24 16th H-Tyr-D-Ser-Gly-Phe-Leu-OCH ₂ I 65 64 54 56 54 40 81 118th Hll. 7, 8, 26 1 CHOAc | 1 CH ₂ OAc 17th H-Tyr-D-Ser-Gly-Phe-Leu-0CH ₂ 67 73 73 55 60 94 119th Hll. 7.8 j CHOCIOKisHai | CH ₂ OC (O) C _{1 S} H ₃₁ 18th H-Tyr-D-Ser-Gly-Phe-Leu-OCH (CH ₂ OAc) ₂ 67 71 65 58 60 44 90 116th Hll. 7, 8, .25 19th H-Tyr-D-Ser-Gly-Phe-Leu -OCHfCHaOCWsHuh 61 68 54 54 51 33 85 117th Hll. 1.12 20th H-Tyr-D-Ser-Gly-Phe-Leu-OBu * 59 76 124th Hl. 4, 14

Continuation of Table 17

i ----------- Example would RfXlO ² Kiin. Mode- Set- material agent Kgy- number A B C D F Η K Q. number ing

21st H - Tyr - D - Thr - Gly - Phe - Leu - OMe 59 47 80 11 130th H9. 1, 13 22nd H-Tyr-D-Trp-Gly-Phe-Leu-OMe; 63 59 85 23 131st H5. 1, 8 23rd H-Tyr-β-Alá-Gly-Phe-Nle-OMe 66 28 8 135th H9. 8 24th Η-β-Ala-Tyr-D-Ala -Gly-Phe-Leu-OMe 62 63 62 82 178th Hl. 1, 17 25th H-Asp-Tyr-D-Ala-Gly-Phe-Leu-OMe 57 58 200th H9. 1, 13 26th H-Lys-Me-Tyr-D-Ser —Gly-Phe-Leu — OMe 29 66 49 209th H5. 2, 16 27th H-Lys-Tyr-D-Ala-Gly- —Phe-DL-NH- (2-Oxo-tetrahydrofur-3-yl) 26 51 22 210th H2. 8 28th H-Lys (Boc) -Tyr-D-Ala-Gly-Phe-Leu-OMe 73 76 77 68 57 55 20 193rd H9. 1, 15 29th H - Tyr - D - Ala - Gly - Phe - Leu - Thr - OH 51 20 218th H2. 7, 8, 27 30th H-Gly-Gly-Tyr-D-Ala —Gly-Phe-Leu-OMe 51 78 55 53 10 48 185th H9. 1, 16 31st H-Asp-Lys-Tyr-D-Ala Gly-Phe-Leu-OMe; 21 25 57 191st H10. 2, 12 32nd H- £ ys H — Lys — Tyr — D — Al— Gly-Phe-Leu-OMe; 7 8 6 32 196th H5. 3, 12 33rd H-Arg-Pro-Lys-Tyr-D-Ala-Gly-Phe-Leu-OMe 58 199th H9. 6, 18 34th Η-Gly-Gly-Gly-Tyr-D-Alla -Gly-Phe-Leu-OMe 65 76 40 49 12 13 88 183rd H9. 1, 13 35th H-Gly-Gly-Gly-Tyr-D-Ser-Gly-Phe (6H) -Leu-OMe 40 69 32 60 65 203rd H9. 4, 18 36th H-Leu-Leu-Leu-Tyr-D-Ala-Gly-Phe-Leu-OMe; 63 60 30 187th Hll. 1, 16 37th H-Olu-Gly-OH H-tys-Tyr-D-Ala-Gly-Phe-Leu-OMe 36 60 27 60 194th H9. 2, 18 38th H-Pro-Gin-Gin-Tyr-D-Alá - Gly-Phe-Leu-OMe 50 217th H9. 4, 12, 28 39th H — Asp — Gly — Gly — Gly — Tyr — D— —Ala-Gly-Phe-Leu-OMe 30 66 19 34 41 180th H9. 1, 16

Continuation of Table 17

Example Sami Product the THE RfXlO ² Kiin. O. number of substance Módiszer Comment B C D F Η K 40th H-Lys-Gly-Gly-Gly-Tyr-D · -Alá-Gly-Phe-Leu-OMe 32 72 4 9 188th H9. 2, 16 41st H-Gly-Gly-Gly H-Lys-Tyr-D-Ala-Gly-Phe-Leu-OMe 28 66 25 42 195th H9. 2, 16, 13 42nd H-Arg-Pro-Lys-Pro-Gln -Gln-Tyr-D-Ala-Gly- 198th H9. 18.19

-Phe-Leu-OMe;

43-69. Example 20

The protected polypeptide used as starting material is treated with hydrochloric acid. The compounds of formula (I) listed in Table 18 are obtained.

The protecting groups are cleaved by the following methods:

Method E: The starting material is dissolved in ethyl acetate and a solution of hydrochloric acid in ethyl acetate is added. The hydrochloric acid concentration is adjusted to 2-6 n ³⁰ . The mixture was allowed to stand at 20-25 ° C for 1-2 h. Use the shortest possible reaction time to eliminate unwanted side reactions. The hydrochloride ³⁵ usually becomes out of solution as a solid. The solid is then isolated by filtration, otherwise the product is isolated by vacuum evaporation of the solution.

E2. Method 4: Method E was used, but methanol was used as the reaction medium.

E3. Method A: Method E1 was carried out, but the reaction medium was 50:50 by volume methanol / ethyl acetate. 45

E4. Method 1: Method E is repeated, but the starting material is dissolved in acetic acid and then a solution of hydrochloric acid in ethyl acetate is added. The final reaction mixture contained acetic acid and ethyl acetate (50: 1 by volume).

E5. Method E2. However, the reaction is carried out under a nitrogen atmosphere in the presence of a cleavage agent (e.g., 2-mercaptoethanol). , ₅₅

E6. Method 1: Proceed to Method E; however, the reaction medium is diethyl ether.

E7. Method I: Method El, except that dioxane is used as the reaction medium.

E8. Method I: Method E is used, but the reaction medium for go is acetic acid.

E9. Method I: Method E1, however, is carried out under a nitrogen atmosphere in the presence of a cleavage agent (e.g. 2-mercaptoethanol). 65

E10. Method 3: The protected polypeptide is stirred at about 10 ml / g in concentrated hydrochloric acid for 10 minutes at 0 ° C. The excess acid was evaporated in vacuo (using the lowest possible temperature) and the product was lyophilized.

Notes to Table 18:

1. Hydrochloric acid salt.

2. Dihydrochloride salt.

3. The mixture is born on diatomaceous earth and evaporated to isolate the product.

4. The product is isolated by precipitation.

5. The product is precipitated at food.

6. The product is precipitated with methanol-ether.

7. The product is crystallized from a 50:50 (v / v) mixture of ethyl acetate and petroleum ether (b.p. 60-80 ° C).

8. The product is lyophilized (medium: water or a 50:50 v / v mixture of water and tert-butanol).

9. See Note 8, the operation is carried out in the presence of hydrochloric acid.

10. Chromatograph the product on silica gel using system Q as solvent.

11. See Note 10, solvent K is used as solvent.

12. Chromatograph the product on a Sephadex G15 gel using 0.01 M aqueous hydrochloric acid as solvent.

13. Chromatograph the product on a Sephadex G25 gel using 5% v / v aqueous acetic acid * as the solvent.

Mp 202-205 ° C.

15. Melting point: 110 ° C.

16. Melting point: 179-180 ° C.

17. Melting point: 173-174 ° C.

Melting point 142-144 ° C.

M.p .: 118-120 ° C.

20. M.p .: 138-140 ° C.

Melting point 111-114 ° C.

M.p .: 195-200 ° C (dec.).

23. M.p .: 120-122 ° C.

M.p. 174-176 ° C (dec).

25. M.p .: 120-124 ° C.

Table 18

examples Rf x 10 ² Kiin. material number Method Comment da sled would C D F H K Q ia A. B 43rd H — Me — Tyr — D — Ser — Gly — Phe— —Leu-OMe 68 77 62 58 44 9 103rd E2. 1, 10 44th H — Me — Tyr — D — SerfBu ¹ ) —Gly— —Phe — Leu-OMe 7 80 73 61 60 28 103rd E2. 1, 10 45th H-Tyr-D-Ala-Gly-Phe -Azleu-NH ₂ 68 79 64 58 41 30 80 145th E8. 1, 4, 14 46th H-Tyr-D-Ala-Gly-Phe-DL-NH (2-oxo-tetrahydrofur-3-yl) 48 65 38 57 67 146th E9. 1, 12 47th H-Tyr-D-Ala-Gly-Phe-Leu-OMe 67 49 21 162nd E7. 1, 5, 15 48th H-Tyr-D-Ala-Gly-Phe-Leu -NH- (cyclohexyl) 67 58 45 87 10 172nd E3. 1, 8, 16 49th H-Tyr-D -Alá-Gly-Phe-Leu - (tetrahydropyrrolo) 65 62 44 96 173rd El. 1, 8, 17 50th H-Tyr-D-Alá-Gly-Phe-Leu - (morpholino) 62 88 174th El. 1, 11, 18 51st H-Tyr-D-Ala-Gly-Phe- —Met — OMe 62 79 61 60 62 54 84 147th E8. 1, 4, 19 52nd H-Tyr-D-Ala-Gly-Phe- —Pro — NHEt 52 72 60 63 73 176th El. 1, 12 53rd H-Tyr-D-Lys-Gly-Phe-Leu-OMe; 54 109th E2. 2, 8 54th H-Tyr-D-Met-Gly-Phe-Leu-OMe 59 49 81 14 148th E5. 1, 7 55th H-Tyr-D-Ser-Gly-Phe -Azleu-NH ₂ 68 80 55 60 50 34 149th E8. 20 56th H-Tyr-D-Ser-Gly-Phe-Leu-OMe 63 76 68 65 88 111th El. 1, 13, 9 57th H-Tyr-D-Ser-Gly-Phe-Leu- (tetrahydropyrrolo) 55 67 57 54 121st El. 1, 8 58th H — Tyr — D Ser-Gly-Phe— —Met-OMe 62 70 67 60 57 53 151st E8. 1, 6, 21 59th H-Glu-Tyr-D-Ala-Gly -Phe-Leu-OMe 58 68 202nd El. 1, 8 60th H-Lys-Tyr-D-Ala-Gly-Phe-AzJeu-NH ₂ 34 65 21 212th E8. 1, 6, 22 61st H-D-Lys-Tyr-D-Ala-Gly-Phe-Leu-OMe; 54 10 65 206th E2. 2, 3 62nd H-Lys-Tyr-D-Ala-Gly-Phe-Leu-OMe 54 68 23 48 72 207th E2. 2, 11

Continuation of Table 18

Example number: Product 1 THE B C RpYlO ² Kiin. substance Q. number Method Comment D E Η Q. 63rd H-Lys-Tyr-D-Ala-Gly-Phe-Met-OMe 33 54 30 51 213th E8. 2, 6, 23 64th H-Lys-Tyr-Azala-Gly-Phe-Leu-OMe; 35 69 21 45 14 42 214th E8. 2, 4, 24 65th H-Lys-Tyr-D Ser-Gly -Phe-Azleu-NH ₂ 38 61 28 50 68 215th E8. 2.4 66th H — Lys — Tyr — D — Ser — Gly— —Phe — Met — OMe 33 54 30 55 216th E8. 2, 5 67th H-Phe H-Lys-Tyr-D-Ala -Gly-Phe-Leu-OMe 60 208th E3. 2, 8 68th H-Gly-Gly-Gly-Me-Tyr-D-Ser-Gly-Phe-Leu-OMe 37 73 25 48 55 205th E10. 1, 11 69th H-Gly-Gly-Gly-Gly-Gly-Gly-Tyr-D-Ala-Gly-Phe-Leu-OMe 31 75 4 13 41 204th E4. 1.4

70-88. example

The protected polypeptide used as starting material is treated with trifluoroacetic acid. The compounds of formula (I) listed in Table 19 are obtained.

The protecting groups are cleaved by the following methods:

FI. Method 1 Dissolve the starting material in trifluoroacetic acid (10 mL trifluoroacetic acid per gram of starting material) and keep at 20-25 ° C for 1-2 h. The solution was concentrated in vacuo. The product is obtained in the form of its trifluoroacetic acid salt. Optionally, the product is freeze-dried in the presence of a small excess of hydrochloric acid. The compound of formula (I) is then obtained as its hydrochloride.

F2. method: FI. However, 50 uses 90% or 95% v / v aqueous trifluoroacetic acid.

F3. method: FI. However, 90 or 95% (v / v) aqueous trifluoroacetic acid is used and the reaction is carried out under a nitrogen atmosphere in the presence of a cleavage agent (e.g., 2-mercaptoethanol).

5. The solution is filtered through diatomaceous earth and the product precipitated from the filtrate.

6. Precipitate the product with petroleum ether.

7. The product is lyophilized (medium: water or a 50:50 v / v mixture of water and tert-butanol).

8. See Note 7, the operation is carried out in the presence of hydrochloric acid.

9. Chromatograph the product on Sephadex G15 gel using 5% aqueous acetic acid as solvent.

10. Melting point: 150-155 ° C.

11. M.p .: 166-168 ° C.

Notes to Table 19: 60

1. Hydrochloric acid salt.

2. Acetic acid salt.

3. Trifluoroacetic acid salt.

4. Bis-trifluoroacetate. 65

Example 89

To a solution of H5-Tyr-D-Ser-Gly-Phe (6H) -Leu-OMe (starting material 110) (235 g, 330 mol) in 5 ml of methanol was added 1.0 ml (3 molar equivalents) of aqueous sodium hydroxide solution ( In) was added and the mixture was stirred at room temperature for 3 hours. The solution was concentrated in vacuo. The residue was diluted with water, acidified with dilute aqueous hydrochloric acid and filtered. The filtrate was freeze-dried. The product was desalted by passing through a column packed with 5% aqueous acetic acid in a Sephadex G15 gel and freeze-drying the filtrate. The product was obtained as H-Tyr-D-Ser-Gly-Phe (6H) ~ Leu-OH, Rf 0.38 (system C).

Table 19

• PEI ² RfxlO K Q Kiin. material number Mode- agent Comment da sled would Iá THE B C D F H 70th H-Tyr-D-Ser-Gly-Phe- (tetrahydropyrazole) 45 58 30 53 81 92nd FI. 4, 5, 10 71st H-Tyr-D-Ala-Gly-Phe-Leu-OH 60 74 161st F2. 3, 7 72nd H - Tyr - D - Ala - Gly - Phe - Leu - O (CH ₂ ) ₂ OH 67 98 163rd F2. 3, 7 73rd H-Tyr-D-Ala-Gly-Phe-Leu-O (CH ₂ ) ₂ NH ₂ 77 164th F2. 3, 7 7.4. H-Tyr-D-Ala-Gly-Phe -Leu-O (GH ₂ ) ₂ NMeZ 81 83 91 78 24 165th F2. 1, 8 75th H - Tyr - D - Ala - Gly - Phe - Leu - OCH ₂ CH = CH ₂ 63 50 91 25 166th F2. 1, 8 76th H-Tyr — D-Ala-Gly-Phe— —Leu — OPh 70 60 20 167th F2. 1, 8 ΊΊ. H - Tyr - D - Ala - Gly - Phe - Leu - NH (CH ₂ ) ₂ OH 64 38 90 168th F2. 3, 7 78th H-Tyr-D-Ala-Gly-Phe-Leu-NH (CH ₂ ) ₂ NHMe 10 71 169th F2. 4, 7 79th H-Tyr-D-Ala-Gly-Phe -Leu-NH (CH ₂ ) ₂ NMe ₂ 170th F2. 4, 7 80th H - Tyr - D - Ala - Gly - Phe - Leu - NEt ₂ 66 53 90 25 171st F2. 3, 7 81st H-Tyr-D-Ala-Gly-Phe -DL-Leu-Me 66 58 11 175th F2. 1, 8 82nd H-Tyr-D-Ser-Gly-Phe-Azpro-NHEt 44 70 43 150th FI. 3, 5, 11 83rd H-Tyr-D-Ser-Gly-Phe-Leu-O (CH ₂ ) ₂ CHMe ₂ 63 39 80 18 120th F2. 3, 6 84th H-Tyr-D-Ser-Gly-Phe-Pro-NHEt 66 44 59 122nd F2. 2, 9 85th H - Tyr - D - Ala - Gly - Phe - Leu - D - Ala - OMe 67 44 219th F2. 3, 7 86th H-Tyr-D-VALA-Gly-Phe- Leu-Ala-OMe 63 52 90 220th F2. 3, 7 87th H-Tyr-D-Ala-Gly-Phe-Leu-Gly-OMe 61 62 94 16 221. · F2. 3, 7 88th H-Tyr-D-Ala-Gly-Phe-Leu-D-Thi-OH 64 77 222nd F2. 3.7

Example 90

Claims (7)

Patent claims: H-Me-Thyr-D-Ser-Gly-Phe (6H) -Leu-OMe (starting material 102) was treated with 2 molar equivalents of sodium hydroxide in aqueous solution and worked up as described in Example 89. H-Me-Tyr-D-Ser-Gly-Phe (6H) -Leu-OH, Rf 0.44 (system C) is obtained. Example 91 A process for preparing the enkephalin analog polypeptides of formula (I) and pharmaceutically acceptable acid addition or base addition salts thereof, wherein: R is hydrogen or methyl, R ² is hydrogen, or acetyl, or Glu, Asp, O-Ala, D-Lys, Lys optionally substituted on the amino group at the e-position by tert-butoxycarbonyl, Ac-Tyr-D-Ala-Gly-Phe-Leu-OMe (starting material 123) (140 mg, 224 μπιόΐ) was dissolved in 10 mL of warm dioxane and allowed to cool to room temperature. 2 ml of water and 0.75 ml (3.5 equivalents) of 1 N aqueous sodium hydroxide solution are added. The mixture was stirred vigorously at room temperature for 2.5 hours and then the bulk of the dioxane was evaporated in vacuo. The residue was acidified with 1N aqueous hydrochloric acid and the resulting suspension was concentrated in vacuo. Aqueous ammonia and 0.05 M aqueous ammonium acetate solution were added to the residue. The resulting clear solution was evaporated to remove excess ammonia and chromatographed on a SephadexG25 gel column. 0.05M aqueous ammonium acetate solution was used as solvent. Fractions containing the desired product were freeze-dried. Ac-Tyr-D-Ala-Gly-Phe-Leu-OH, Rf 0.56 and 0.47 (system D) are obtained. Example 92 A solution of 100 mg (157 pmoles) of H-Me-Tyr-D-Al-Gly-Phe-Leu-OMe · HCl (product of Example 1) in 15 ml of methanol was cooled to 4 ° C and saturated with anhydrous ammonia. . After 2 days at 4 ° C, the solvent was evaporated in vacuo and the residue was chromatographed on a silica gel column. The eluent is QL and then K. The fractions containing the desired product were evaporated and the residue was freeze-dried. The product was a H-Me-Tyr-D-Ala-Gly-Phe-Leu-NH ₂ as a colorless oil, _Rf: 0.22 and 0.70 (H. K. system). H-Glu-Gly-OH Η-Lys-, Η-Phe H-Lys-, H-Gly-Gly-Gly-Lys-, H-Asp H-Lys-, H-Lys H-Lys-, Η-Gly-Gly-, H-Leu-Leu-Leu-, Η-Arg-Pro-Lys-, H-Pro-Gln-Gln-, Η-Gly-Gly-Gly- , H-Lys-Gly-Gly-Gly, Η-Asp-Gly-Gly-Gly, H-Aig-Pro-Lys-Pro-Gin-Gin, or Η-Gly-Gly-Gly-Gly-Gly-Gly, B is D-Al, β-Al or Azala, E is Gly or Azgly, F is Phe, hexahydroPhe or Azphe, G is DL-Leu, Leu, D-Leu, Azleu, Nle, Met, Pro or Azpro, and K is hydroxy, amino, phenoxy, C 1-6 alkoxy, C 2-4 alkenyloxy, hydroxy, amino, C 1-4 alkflamino, N- (1-4C) C 1-4 alkoxy, C 1-4 alkoxy, C 1-4 alkoxy, C 1-4 alkanoyl, or C 1-20 alkanoyloxy, or two C 1-20 alkanoyloxy substituents or two C 1-20 alkanoyloxy substituents, C 1-4 alkylamino, C 5-7 cycloalkylamino groups. , C1-C4 alkylamino, di- (C1-C4) alkylamino, hydroxy, C1-C4 alkylamino, or di (C1-C4) alkylamino, morpholino a tetrahydropyrrolo group or a (1-4C) alkyl group, or G and K together form a group of formula (III) or (IV) in the D, L, or DL configuration, or a group of formula (II), provided that at least one of B, E, F and G is represents a residue other than an amino acid residue, or Example 93 To a solution of 100 mg of H-Tyr-D-Ala-Gly-Phe-OMe in 1 ml of dimethylformamide was added 50 molar equivalents of ethylamine and the mixture was stirred for 3 days at 4 ° C. The solvent was evaporated in vacuo and the residue was chromatographed on a silica gel column. Elution system K is used. The fractions containing the desired product were evaporated and the residue was freeze-dried. The product obtained was H-Tyr-D-Ala-Gly-Phe-Leu-NHEt, Rf 0.62, 0.50 and 0.90 (D., H. and K-system). ₆₅ R ¹ and R ² are both hydrogen, K is C 1-6 alkoxy, G is Leu, D-Leu, Azleu or Nle, E is Gly or Azgly, F is Phe, hexahydroPhe or Azphe, and B is D-Asp, D-Thr, D-Trp, D-Leu, D-Lys or D-Met, with the proviso that at least one of E, F and G is other than an aza-amino acid residue, or (in) R ^{1 is} hydrogen, R ² is hydrogen, D-Lys, or Lys ⁵ optionally substituted on the amino group at the e-position by a t-butoxycarbonyl group, B is a D-Ser group optionally substituted on the hydroxy group at the 0-position by a t-butyl group, K is a C1-6 alkoxy group, ¹⁰ E is Gly or Azgly, F is Phe, hexahydroPhe or Azphe, and G is a Met group, provided that E and F are at least one other than one of the ¹⁵ aza-amino acid residues, or (iv) R ^l and R ² are both hydrogen, ²⁰ K is an amino group optionally substituted with one or two C 1 -C 4 alld1 groups, B is D-Ser ²⁵ optionally substituted on the hydroxy group at the 0-position by a t-butyl group, E is Gly, F is Phe, and G is Pro or Azpro, or (v) R ¹ is hydrogen or methyl, E Gly means a group, F Phe hexahidroPhe or Azphe refers to a group of 35 G is Leu, D-Leu, Azleu or Nle, R ² is hydrogen, D-Lys groups, where the e-position amino group substituted with t-butoxycarbonyl groups of a Lys residue or H-Gly-Gly-Gly, 40 K represents a hydroxy group, an amino group, a C 1-6 alkoxy group, a C 1-4 alkoxy group substituted by one hydroxy or C 1-20 alkanoyloxy group or two C 1-20 alkanoyloxy groups or a tetrahydropyrrolo group; group, and B is a D-Ser group optionally substituted on the hydroxy group at the 0-position by a t-butyl group, or (vi) R ¹ is hydrogen or methyl, R ² is hydrogen or acetyl, B is D-Ala or 0-Ala, E is Gly group 55 F is Phe or hexahydroPhe, G is Leu or D-Leu, and K is Gly-OMe, Thr-OH, D-Thr-OH, Ala-OMe or D-Ala-OMe, and the amino acids listed are 50 L-configuration, unless otherwise indicated, (a) deprotection of the corresponding protected polypeptides; or ¢ 5 (b) for the preparation of compounds of formula (I) other than hydroxy in which K is substituted in (i) or (vi) and for the preparation of compounds of formula (I) in which G and K together have the formula (II) reacting a peptide of formula (V) or (VI) wherein R ¹ , R ² , Β, E, F and G, or a reactive derivative thereof, with the appropriate alcohol or amine, and optionally forming a salt thereof. The compounds of formula (I) are converted into their pharmaceutically acceptable acid addition salts and their pharmaceutically acceptable base addition salts. (Priority: April 6, 1977) The process of claim 1 for the preparation of enkephalin analog polypeptides of formula (I) and pharmaceutically acceptable acid addition or base addition salts thereof, wherein (O ¹ R ¹ is hydrogen or methyl, R ² is hydrogen) or R ^{1 is} hydrogen, R ^{2 is} acetyl, Glu, Asp, O-Ala, D-Lys, Lys, H-Glu-Gly-OH Η-tys-, Η-Phe H-Lys-, H-Gly-Gly-Gly Η-Lys-, Η-Asp H-Lys-, H-Lys Η-Lys-, H-Gly-Gly-, H-Leu-Leu-Leu-, H-Arg-Pro-Lys-, H-Pro-Gin-Gin-, H-Gly-Gly-Gly-, Η-Lys-Gly-Gly-Gly-, H-Asp -Gly-Gly-Gly-, H-Arg-Pro-Lys-Pro-Gin-Gin, or H-Gly-Gly-Gly-Gly-Gly-Gly B is D-Ala or Azala, E is Gly or Azgly, F is Phe, hexahydroPhe or Azphe, G is Leu, D-Leu, Azleu, Nle, Met, Pro or Azpro, and K is hydroxy, amino, phenoxy, C 1-6 alkoxy, C2-C4 alkenyloxy, C1-C4 alkoxy substituted with one hydroxy, amino, C1-C4 alkylamino or C1-C20 alkanoyloxy, or with two C1-C20 alkanoyloxy groups, C 4 -alkylamino, di-C 1-4 -alkylamino, tetrahydropyrrolo, or morpholino, or G and K together form a group of formula (ΙΠ) or formula IV or a group of formula (D) in the D or L configuration, provided that at least one of Β, E, F and G is different from the aza-amino acid residue represents a group, or R ¹ and R ² are each hydrogen, K is C 1-6 alkoxy, G is Leu, D-Leu, Azleu or Nle, E is Gly or Azgly, F is Phe, hexahydroPhe or Azphe, and B D is Asp, D-Thr, D-Lys, D-Trp or D-Met, provided that at least one of E, F and G is other than an aza-amino acid residue, or (ii) R ^{1 is} hydrogen, R ² is hydrogen, D-Lys or Lys, B is a D-Ser group optionally substituted on the hydroxyl group at the β-position by a tert-butyl group, K is C 1 -C 6 alkoxy, E is Gly or Azgly, F is Phe, hexahydroPhe or Azphe, and G is a Met group, provided that at least one of E and F is other than an aza-amino acid residue, or (iv) R ¹ and R ² are both hydrogen, K is an amino group optionally substituted with one or two C 1-4 alkyl groups, B is a D-Ser group optionally substituted on the hydroxy group at the β-position by a t-butyl group, E is Gly, F is Phe, and G is Pro or Azpro, or (v) R ¹ is hydrogen or methyl, E Gly means a group, F Phe, Azphe hexahidroPhe or represents a group G Leu, D-Leu, Nle Azleu or, one of R ² is hydrogen or D-Lys, Lys or H-Gly-Gly-Gly-group., K is a hydroxy group, an amino group, a C 1-6 alkoxy group, a C 1-4 alkoxy group substituted by one hydroxy or C 1-20 alkanoyloxy group or two C 1-20 alkanoyloxy groups or a tetrahydropyrrolo group. , and B represents a D-Ser group optionally substituted on the hydroxy group at the β-position, and the amino acids listed are of the L-configuration unless otherwise indicated, (a) cleavage of protecting groups of the corresponding protected polypeptides; or (b) for the preparation of compounds of general formula (I) other than hydroxy in which K is substituted with the substituents listed in (1) and compounds of general formula (I) in which G and K together form a group of formula (II), reacting a peptide of formula (V) or (VI), wherein R ¹ , R ² , Β, E, F, and G are as defined above, or a reactive derivative thereof with the appropriate alcohol or amine and optionally a salt-forming general formula (I); The compounds of formula Ia are converted into their pharmaceutically acceptable acid addition salts and their pharmaceutically acceptable base addition salts. (Priority: October 28, 1976) 3. The process of claim 1 for the preparation of enkephalin analog polypeptides of formula (I) and pharmaceutically acceptable acid addition or base addition salts thereof, wherein (i) R ^{1 is} hydrogen or methyl, R ² is hydrogen, or when R ^{1 is} hydrogen, R ^{2 is} acetyl or Glu, Asp, β-Ala, D-Lys, Lys, H-Glu-Gly-OH t-tys-, H-Éhe H-Lys-, H-Gly-Gly-Gly Η-Lys-, H-Asp H-Lys- or «----------------- 1 H-Lys may also represent H-Lys, B is D-Ala or Azala, E is Gly or Azgly, F is Phe or Azphe, G is Leu, Azleu, Nle, Met or Azpro, and K is a hydroxy group, an amino group, a C 1-6 alkoxy group, a C 2-4 alkenyloxy group, a phenoxy group, a C 1-4 alkoxy group substituted by a hydroxy, amino or C 1-4 alkylamino group. C 1-4 alkylamino, di- (C 1-4) alkylamino, tetrahydropyrrolo or morpholino, provided that at least one of Β, E, F and G is represents a residue other than an amino acid residue, or R ¹ and R ² are each hydrogen, K is C 1 -C 6 alkoxy, G is Leu, Azleu or Nle, E is Gly or Azgly, F is Phe or Azphe, and B is D-Thr, provided that at least one of E, F and G is other than an aza-amino acid residue, or (iii) R ^{1 is} hydrogen, R ² is D-Lys or Lys, B is a D-Ser group optionally substituted on the hydroxy group at the β-position by a t-butyl group, K is C 1 -C 6 alkony, E is Gly or Azgly, F is Phe or Azphe, and G is Met, provided that at least one of E and F is other than an aza-amino acid residue. , or (iv) R ¹ and R ² are both hydrogen, K is an amino group optionally substituted with one or two C 1 -C 4 alkyl groups, B is a D-Ser group optionally substituted on the hydroxy group at the N-position by a t-butyl group, E is Gly, F is a Phe group and G is an Azpro group, or (v) R ^{1 is} hydrogen or methyl, E Gly means a group, F represents Phe or Azphe group, G is Leu, Azleu or Nle, R ² is hydrogen, D is Lys or Lys, K is hydroxy, amino, C 1-6 alkoxy, hydroxy (C 1-4) alkoxy or tetrahydropyrrolo, and B represents a D-Ser moiety optionally substituted on the hydroxy group at the 0-position and the amino acids listed are L-configurations unless otherwise indicated, (a) cleavage of protecting groups of the corresponding protected polypeptides; or b) for the preparation of compounds of formula (I) other than the hydroxyl group of the substituents listed in (i), a peptide of formula (V) or (VI) - wherein R ¹ , R ² , B, E, F and G is a reacting derivative of the above or a reactive derivative thereof with the appropriate alcohol or amine and, if desired, converting the compounds of formula (I) into a pharmaceutically acceptable acid addition salt or a pharmaceutically acceptable addition salt thereof with a base. (Priority: July 14, 1976) The process of claim 1 for the preparation of enkephalin analog polypeptides of formula (I) and pharmaceutically acceptable acid addition or base addition salts thereof, wherein (i) R ¹ is hydrogen or methyl, R ² is hydrogen, or when R ^{1 is} hydrogen, R ^{2 is} acetyl, Glu, Asp, β-Ala, D-Lys, Lys, H-Glu-Gly-OH H-Lys-, H-Phe H-Lys-, H-Gly-Gly-Gly Η-Lys-, Η-Asp H-Lys ¹ ----------- ----- ¹ or H-Lys can be H-Lys, B is D-Sub, E is Gly, F is Phe, G is Leu, Met or Nle, and K is hydroxy, amino, phenoxy, C 1-6 alkoxy, C 2-4 alkenyloxy, C 1-4 alkoxy substituted with hydroxy, amino or C 1-4 alkylamino, C 1-4 alkylamino, di (C 1-4) alkylamino , tetrahydropyrrolo, or morpholino. or (ii) R ¹ and R ² are each hydrogen, K is C 1 -C 6 alkoxy, G is Leu or Nle, E is Gly, F is Phe, and B represents a D-Thr group, or (iii) R ^{1 is} hydrogen, R ² is hydrogen or D-Lys or Lys, B is a D-Ser group optionally substituted on the hydroxyl group at the β-position by a tert-butyl group, K is C 1-6 alkoxy, E is Gly, F is Phe, and G represents a Met group, or (iv) R ¹ is hydrogen or methyl, E Gly means a group, F stands for Phe, G is Leu or Nle, R ² is hydrogen, D-Lys or Lys, K is hydroxy, amino, C 1-6 alkoxy, hydroxy (C 1-4) alkoxy or tetrahydropyrrolo, and B represents a D-Ser moiety optionally substituted on the hydroxy group at the β-position and has the L-configuration, unless otherwise indicated, characterized in that (a) cleavage of protecting groups of the corresponding protected polypeptides; or b) for the preparation of a compound of formula (I) other than the hydroxyl group of the substituent listed in (i), a peptide of formula (V) or (VI) - wherein R ¹ , R, B, E, F and G or a reactive derivative thereof, or a reactive derivative thereof, with the appropriate alcohol or amine and, if desired, converting the compounds of formula (I) into a pharmaceutically acceptable acid addition salt or a pharmaceutically acceptable addition salt with bases. (Priority: May 27, 1976) The process of claim 1 for the preparation of enkephalin analog polypeptides of formula (I) and pharmaceutically acceptable acid addition or base addition salts thereof wherein: R ¹ is hydrogen, R ² is hydrogen or Glu, Asp, β-Ala, D-Lys, Lys, H-Glu-Gly-OH Η-Éys-, Η-Phe H-tys-, H-Gly-Gly-Gly-ys-Lys, Η-Asp H-Lys »'1, or H-Lys H-Lys, B is D-Ala or Azala, E is Gly or Azgly, F is Phe or Azphe, G is Leu, Met, Nle, Azleu or Azpro, and K is hydroxy, amino, phenoxy, C 1-6 alkoxy, C 2-4 alkenyloxy, C 1-4 alkoxy substituted with hydroxy, amino or C 1-4 alkylamino, C 1-4 alkylamino, di- (C 1-4) alkyl- an amino group, a tetrahydropyrrolo group, or a morpholino group, provided that at least one of B, E, F and G is other than an aza-amino acid residue, or (ü) R ¹ and R ² are each hydrogen, K is C 1 -C 6 alkoxy, G is Leu, Azleu or Nle, E is Gly or Azgly, F is Phe or Azphe, and BD is Thr, with provided that at least one of G, E and F is other than an aza-amino acid residue, or (iii) R ^{1 is} hydrogen, R ² is hydrogen, D-Lys or Lys, B is a D-Ser group optionally substituted on the hydroxy group at the N-position by a t-butyl group, K is C 1 -C 6 alkoxy, E is Gly or Azgly, F is Phe or Azphe, and G is Met, provided that at least one of E and F is other than an aza-amino acid residue, or (iv) R ¹ and R ² are both hydrogen, K is an amino group optionally substituted with one or two C 1 -C 4 alkyl groups, B is a D-Ser group optionally substituted on the hydroxy group at the β-position by a t-butyl group, E is Gly, F is Phe, and G is an Azpro group, or (v) R ^{1 is} hydrogen, E Gly means a group, F represents Phe or Azphe group, G is Leu, Azleu or Nle, R ² is hydrogen, D-Lys or Lys, K is hydroxy, amino, C 1-6 alkoxy, hydroxy (C 1-4) alkoxy or tetrahydropyrrolo, and B represents a D-Ser moiety optionally substituted on the hydroxy group at the β-position and has the L-configuration, unless otherwise indicated, characterized in that (a) cleavage of protecting groups of the corresponding protected polypeptides; or b) for the preparation of compounds of formula (I) other than the hydroxy group in which K is a substituent as defined in (i), a peptide of formula (V) or (VI) - wherein R ¹ , R ² , B, E, F and G is a reacting derivative of the above or a reactive derivative thereof with the appropriate alcohol or amine and, if desired, converting the compounds of formula (I) into a pharmaceutically acceptable acid addition salt or a pharmaceutically acceptable addition salt thereof with a base. (Priority: May 21, 1976) 6. The process of claim 1 for the preparation of enkephalin analog polypeptides of formula (I) and pharmaceutically acceptable acid addition or base addition salts thereof wherein: R ¹ is hydrogen, R ² is hydrogen or Glu, Asp, β-Ala, D-Lys, Lys, H-Glu-Gly-OH Η-Lys-, Η-Phe H-Lys-, Η-Gly — Gly-Gly H-ly, Η-Asp H-Lys, or r ...... · H-Lys H-Lys, B is D-Ala, E is Gly, F is Phe, G is Leu, Met or Nle, and K is hydroxy, amino, phenoxy, C 1-6 alkoxy, C 2 -C 4 alkenyloxy, C 1 -C 4 alkoxy substituted with hydroxy, amino or C 1 -C 4 alkylamino, C 1 -C 4 alkylamino, di- (C 1 -C 4) - an alkylamino group, a tetrahydropyrrolo group, or a morpholino group, or (ü) ₂ R * and R ² are each hydrogen; K is C 1 -C 6 alkoxy, G Leu or Nle group, meaning E is Gly, F is Phe, and B represents a group D-Thr, or 20 (iii) R ^{1 is} hydrogen, R ² is hydrogen or Lys or D-Lys 25, B is a D-Ser group optionally substituted on the hydroxy group at the β-position by a t-butyl group, K is C 1 -C 6 alkoxy, E Gly means a group, F is a Phe group, and G stands for Met, or (arc) R ¹ is hydrogen, E Gly means a group, F stands for Phe, G is Leu or Nle, R ² is hydrogen or Lys or D-Lys, K is hydroxy, amino, C 1-6 alkoxy, hydroxy (C 1-4) alkoxy or tetrahydropyrrolo, and B represents a D-Ser moiety optionally substituted on the hydroxy group at the β-position and has the L-configuration, unless otherwise indicated, characterized in that (a) cleavage of protecting groups of the corresponding protected polypeptides; or b) for the preparation of a compound of formula (I) other than a hydroxy group, wherein K is a substituent listed in (i), a peptide of formula (V) or (VI) - wherein R ¹ , R ² , Β, E, F and G is as defined above, or a reactive derivative thereof, is reacted with the appropriate alcohol or amine and, if desired, the compounds of formula (I) suitable for the salt formation are converted into their pharmaceutically acceptable acid addition salts or their pharmaceutically acceptable addition salts. (Priority: April 8, 1976) 7. A process for the preparation of a pharmaceutical composition according to claim 1, characterized in that an encephalin analogue polypeptide of formula (I) - wherein R ¹ , R ² , Β, E, F, G and K are A pharmaceutically acceptable acid addition or base addition salt thereof as defined in claim 1 or a pharmaceutically acceptable salt, diluent and / or excipient thereof, in a known manner, in admixture with conventional pharmaceutical carriers, diluents and / or excipients. (Priority: April 6, 1977) 1 drawing Responsible for publishing: Director of Economic and Legal Publishing 814081 - Zrínyi Printing House, Budapest 176260 International Classification: C 07 C 103/52