HU184704B - Process for preparing biologically active encephaline analogues - Google Patents

Abstract

A peptide of the formula I is prepared by reacting two compounds which, in a complementary manner, give the desired peptide chain. The symbols in Formula I have the meaning given in Patent Claim 1. The compounds of the formula I exhibit interesting physiological properties, in particular an activity as morphine antagonists. R-(X<2>)n-X<3>-X<4>-X<5>-X<6>-X<7>-(X<8>)p-(X<9>) q-R<1> (I)

Description

This invention relates to novel peptides and derivatives useful in human and veterinary medicine, and to pharmaceutical compositions containing them as active ingredients.

More particularly, the present invention relates to the production of morphine agonist peptides and derivatives. Commonly referred to as morphine agonists are compounds whose biological activity mimics that of natural alkaloids. .

The pharmacological properties and therapeutic uses of morphine can be found, for example, in The Pharmacological Basis of Therapeutics, Goodman, L.S., and Gilman, A, eds. label: The MacMillan Company, New York,

3rd edition / 1965 / in particular Chapter 15, pp. 247-266. p. and Martindale: The Pharmaeeutical Press Lón Don, 26th Edition (1972), especially 1100-1106. p. However, it is known (Goodman, L.S. et al., Loc. Cit., Chapter 16) that repeated administration of morphine may result in the patient becoming accustomed to morphine, becoming accustomed to its effects, and withdrawal symptoms when accompanied by withdrawal symptoms.

Therefore, research has been going on for several years to produce a compound which shows the effects of morphine without having harmful properties.

The present invention provides novel peptides of the formula I as well as their salts, C 1 -C 4 alkyl esters, amides, N 1 -C 4 alkyl amides and acid addition salts, which are morphine agonists both in vivo and in vitro. . In formula I, n is 0 or 1,

X ¹ is L-arginyl,

X ³ is (D or L) -tyrosyl, (D or L) -N-methyltyrosyl, (D or LV 3, ⁴ -dihydroxyphenylalanyl, L 4'-methyltyrosyl, LO-acetyltyrosyl, L-phenylalanyl -, L-4-chlorophenylalanyl or L- / 3-homothyrosyl,

X ⁴ is glycyl, sarcosyl, a-methylalanyl. (D or L) -alanyl, (D or L) -N-methyl-alanyl, L-asparaginyl, L-isoleucyl, L-prolyl, D-leucyl or D-tryptophyl,

^X5 is glycyl, sarcosyl, (D or L) -alanyl-, or L-prolyl aszparaginilcsoport,

X * is (D or L) -phenylalanyl, L-tyrosyl, LO ^4' -methyl-tyrosyl, L-4-chlorophenylalanyl, LC-phenylglycyl, L-leucyl, Lmethionyl, or L-histidyl .

X ⁷ is glycyl or (D or L) leucyl, (D or L 1 -methionyl, L-alanyl, L-isoleucyl, L-norleucyl, Lvalyl, Lmethionyl sulfoxide, L-threonyl , L-prolyl or D-O homoleucyl,

X * is (D or L) -threonyl, L-lysyl, L-phenylalanyl or lithyrosyl,

X ⁹ is glycyl or L-lysyl,

R is hydrogen or (1-4C) alkyl

R * is the hydroxy group of the 1-carboxyl group of the C-terminal amino acid group, or the group -CH ₂ OR ⁴ which is substituted by the 1-carboxyl group, where R ⁴ is hydrogen or C 1 -C 4 alkanoyl, or 5-tetrazolyl and p and q is 0 or 1 with the exception of compounds of formula V and R-LPhe-LBe-Gly-LLeu-LMet-OH and salts thereof, C 1 -C 4 alkyl esters, amides, N 1 -C 4 alkyl amides and acid addition salts thereof. salts where R is as defined above, X ⁷ is L-leucyl or L-methionyl and X ³ is either L-tyrosyl and then X * is L-phenylalanyl or X ³ is lithyrosyl and X ⁶ is L 4 -chlorophenylalanyl.

Abbreviations used for amino acids are known in the literature and can be found in: Biochemistry 11, 1726 (1972), Except for glycine and unless specifically mentioned below, they will always be referred to as L-configuration amino acids and their derivatives.

When the phenyl group in the X ³ group contains 1 or 2 µg substituents, these substituents are preferably in the 3 and / or 4 position of the ring.

When R * is -CH ₂ OR ⁴ - wherein R ^{4 is} alkanoyl - the alkyl group preferably contains from 1 to 4, preferably from 1 to 2 carbon atoms.

Among the esters of the peptides of formula 1, alkyl esters may be mentioned. Alkyl esters include those having from 1 to 4 carbon atoms in the alkyl group, such as methyl, ethyl, t-butyl.

The peptides of formula I are Ν-alkyl, the alkyl 25 groups having 1 to 4 carbon atoms. Amides of peptides include, in addition to amides derived from ammonia, pyrrolidinamides, piperidinamides and moronic folinamides derived from heterocyclic bases such as pyrrolidine, peperidine and morpholine.

The N-alkyl amides of the peptides, as a subclass of the compounds of formula 1, are characterized by the compound of formula VI - wherein X ² , X ³ , X ⁴ , X ^s , X, X ⁷ , X, X ⁹ , n, p, q and R is as defined above and R ⁵ and R ⁶ and the nitrogen atom to which they are kapcsolód35 form discrepancies, including amino, pyrrolidino, piperidino, morfoΙϊηο-, N-alkylamino - wherein alkyl is 1-4 carbon atoms.

In the case of the acid addition salts and derivatives of the peptides of the formula I, the effect is based on the base and the acid is less important, although pharmacologically and pharmacologically acceptable salts may be used for therapeutic purposes. Examples include acids

(a) mineral acids: hydrochloric, hydrobromic, phosphoric, metaphosphoric, nitric, sulfuric,. b) organic acids: tartaric acid, acetic acid, citric acid, malic acid, lactic acid, fumaric acid, benzoic acid, glycolic acid, gluconic acid, gulonic acid, succinic acid, arylsulfonic acid such as p-toluenesulfonic acid. Pharmaceutically and pharmacologically acceptable acid addition salts and unacceptable salts such as hydrofluoride and gQ perchloric acid salts are useful in isolation and base purification and, of course, unacceptable salts may also be useful in the preparation of useful salts by methods known per se.

Peptides having multiple free amino groups and their derivatives may be prepared as mono- or poly-acid addition salts or as mixed salts of a variety of acids.

• In peptide salts (the peptide consists of a carboxylate amide and a cation), the quality of the cation is less important, although only pharmacologically and pharmaceutically acceptable salts can be used for therapeutic purposes. Examples of suitable cations are sodium and potassium.

Morphinagonist properties of the peptides of formula I and their derivatives are described below without limitation.

A) In vitro:

(a) Inhibition of nerve-induced constriction of iron deferent mice in isolated mice was investigated by the method of Hughe et al. (Brain Researc, 88, 296 (1975)) (pulse at 0.1 Hz) and inhibition by a known narcotic antagonist, naxolone], -allyl-7,8-dihydro-14-hydroxy-normorphinone) was eliminated.

(b) Reduction of electrically induced contractions in isolated guinea pig ileum if, according to Patron [Brit. J. Pharmacol. 12, 119-127 (1957)). Each intestine was drilled through the anode and resuspended in a dose of 2-3 g, Inger parameters:

frequency: 0.1 Hz, duration: 0.4 ms, voltage (supramaximal 30-40 V, contractions were isotonic transformed).

(B) In carrier:

a) The compounds exhibit analgesic activity, for example, in reducing the seizures induced by phenylbenzoquinone in mice when Hendershot et al. Pharm. exp. Therap., 125, 237 (1959)] (the compounds were administered by intracerebroventricular injection) and this decrease was reversed by naxolone. '

b) the compounds also exhibit anti-cough activity, for example if Boura et al., Brit. J. Pharmacol. 39, 225 (1970) in guinea pig.

c) the compounds also exhibit anti-diarrheal activity, for example, in reducing rat castor oil diarrhea.

A narrower range of compounds of formula I are those wherein p and q are 0,

X ^{3 is} L-tyrosyl,

X ⁴ D-alanine group,

X ^! glycyl,

X ⁴ phenylphenyl group,

X ^{7 is} D-leucyl or D-methionyl.

A further subgroup of peptides of Formula 1 is represented by Formula VII, wherein

R, R *. P. 4 has the same meaning as in formula I,

X ^{3 is} L-tyrosyl, O-acetyl-L-tyrosyl or N-methyl-L-tyrosyl,

X ^{4 is} glycyl, uranyl, α-methylalanyl or D-alanyl,

X ^{5 is a} glycyl group, a sarpzyl group or an L-asparaginyl group,

X ^{6 is} L-phenylalanyl, L-tyrosyl or L-4-chloro-phenylalanyl,

X ⁷ Lleucyl, D-leucyl, L-methionyl, D-methionyl, Unorleucyl, bthreonyl or D-u-homoleucyl,

X * bthreonyl, D-threonyl, L-phenylalanyl, L-tyrosyl or L-lysyl, and

X 'is a glycyl or L-lysyl group.

A further narrower range of compounds of Formula I is that of Formula VIII, wherein

RR ¹ is as defined for formula I,

X 'is L-tyrosyl or N-methyl-L-tyrosyl and

X ^{7 is} D-leucyl or D-methionyl.

Peptides of Formula I and Derivatives thereof

the azole substituents have the meanings given above, may be prepared by any of the known methods for the preparation of compounds of similar structure. For example, the corresponding amino acids may be prepared by sequential linkage, the classical method of peptide synthesis, or solid phase methods, or first peptide subunits may be prepared and then subunits linked.

In carrying out such reactions, for example, the carboxyl group of the amino acid is activated and unreacted amino and carboxylic acid groups are protected. This method is known in the peptide literature. By way of example, the following literature references include appropriate activation and deprotection reactions and reaction conditions (including coupling and deprotection reactions).

a) 1.042.487, 1.048.086. and 1,281,383. s. British Patents.

b) Schroder and Lüebke, The Peptides (Academic Press) / 1965 /.

c) Bellean and Malek, J. Am. Chem. Soc.

d) Tilak, Tetrahedron Letters, 849/1970 /.

e) Beyerman, Helv. Chim. Acta., 56, 1729 (1973).

f) Stewart and Young, "Solid Phase Peptide Synthesis" (W. H. Freeman and Co.) / 1969 /.

Depending on the reaction conditions, the peptides of formula I and their derivatives are obtained in the form of the free base or salts thereof or, in the case of peptides, in the form of salts. The acid addition salts can be converted into the free base or salts of other acids and the bases into the acid addition salts by methods well known in the art. Similarly, peptides may be converted into their salts and salts may be converted into peptides or other salts by known methods.

The peptides of formula I and their derivatives and acid addition salts thereof can be prepared by reacting a compound of formula II wherein R is as defined in formula I and Y ¹ is an N-terminal radical fragment of formula I with a compound of formula III ^Y2 is H of formula A compound gyökfragmensét general, additional compounds of formula I the C-terminal gyökfragmens - condensed, and optionally II and compounds of formula III are protected and / or activated and optionally removed from the product deprotected and the product base or a salt, or to an acid addition salt.

For peptides of formula I and derivatives thereof wherein X ^{7 is} (p or L) methionyl, the compound of formula II as defined above preferably corresponds to the N-terminal moiety of compound I containing either X ⁷ at the C-terminal position and then III or contain the group X ⁴ is at the C-terminal position, and then the reagent of formula III includes X ⁷ methionyl residue at the N-terminal position groups compound the N-terminal position (X ^8); Generally, the former is preferred.

It will be clear to those skilled in the art that

184,704 arginyl and the (D or L) honioarginyl (Har) groups can not only be incorporated into the peptide chain as described above, but may also be formed in situ in the fragment already formed by an (D or L) omityl or (D or L) lysyl group. guanidálásával. The reagent used is 1-guanyl-3,5-dimethylpyrazole.

It will also be apparent to those skilled in the art that the peptides of formula I and their derivatives may be converted by other in situ methods. For example, amides, N-C 1-4 alkylamides, for example, may be prepared by reacting a peptide alkyl ester, such as methyl ester, with ammonia, a heterocyclic base or monoalkylamine. Peptide esters can be prepared by standard esterification procedures and the esters are converted to peptides by saponification. All of these processes are standard methods in peptide chemistry and are described in the literature for the formation and removal of reaction conditions and protecting groups. The C 1 -C 4 alkyl esters, amides, N 1 -C 4 alkyl amides, and pharmacologically and pharmaceutically acceptable acid addition salts of the peptides of formula 1, their pharmacologically and pharmaceutically acceptable salts, are useful in any human or veterinary medicine due to their morphine agonist activity. where morphine effect is indicated.

Specially used in the following areas of impact:

1. Pain relief (analgesia), such as the elimination of pain from smooth muscle spasms, the reduction of pain in kidney and gallstones, terminal diseases such as cancer, post-operative pain, and pain in childbirth.

2. Relaxation, for example as a preanesthetic, sedative, initiation of sleep, especially if insomnia is a result of pain or cough and anxiety in general.

3. Suppression of cough.

4. Removal of unsightly breathing, such as left ventricular dysfunction or pulmonary edema.

5. Induction of constipation, for example after ileostomy or colostomy and in the treatment of diarrhea or dysentery.

6. Inducing euphoria and treating depression, for example, by reducing pain caused by a terminal illness such as cancer.

The amount of peptide or derivative or acid addition salt thereof (hereinafter referred to as "the active ingredient") in the various applications will vary depending on the route of administration and the severity of the condition being treated and will depend on the judgment of the physician or veterinarian. Generally, for all uses, the dose will be 0.0025 Mg to 40 mg (ranging from kg to mammal body weight, preferably 0.25 to 400 pg / kg) based on the peptide base.

The active ingredient may be administered by any route appropriate to the condition being treated, such as orally, rectally, nasally, topically (face), vaginally, and parenterally (i.e., subcutaneously, intramuscularly and intravenously). It will be appreciated that the preferred route of administration may be selected according to the condition being treated, for example, administration to the spinal cord may be advantageous to reduce birth pain. Although it is possible to administer the pure product, it is still preferred to prepare a pharmaceutical composition containing the active ingredient.

In addition to the active ingredient, the compositions of the present invention comprise one or more carriers and optionally other pharmaceutical components. The carriers must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof. Preferably, the formulations do not contain oxidizing agents and other substances known to be incompatible with peptides.

The formulations are suitable for oral, rectal, nasal, topical (facial) vaginal, parenteral (i.e., subcutaneous, intramuscular, and intravenous) administration, although the most preferred route of administration will depend upon the active ingredient and the condition being treated. The compositions may be presented in unit dosage form in any manner known in the art. Each method is the same as contacting the active ingredient with a vehicle consisting of one or more excipients. In general, the active ingredient is admixed with a liquid carrier or a finely divided solid carrier, or both, and optionally formulated into the desired composition. Oral compositions of the present invention may be in the form of capsules, wafers or tablets, each containing a predetermined amount of the active ingredient, or as a powder or granules, or as a solution or suspension in an aqueous or non-aqueous liquid, an oil in water in the form of an emulsion. The active ingredient may also be formulated as a pill or electuarium or as a paste.

The tablet may be prepared by compression or compression, optionally with one or more accessory ingredients. Compressed tablets may be made by a suitable machine, and the active ingredient may be in a fluid form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, lubricant, surfactant or dispersant. Compressed tablets are also molded using a suitable machine, and the powdered mixture is moistened with an inert liquid diluent.

Formulations for rectal administration are prepared in the form of a suppository, mixed with conventional carriers such as cocoa butter, and formulated as nasal drops for nasal administration, containing the active ingredient in the form of an aqueous or oily solution.

Formulations suitable for topical oral administration qq consist of lozenges comprising the active ingredient in a flavored form, preferably in the form of sucrose, acacia or tragacanth, or in the form of lozenges containing the active ingredient in an inert base such as gelatin and glycerol or sucrose and acacia.

For vaginal administration, pessaries, creams, pastes, or sprays containing not only the active ingredient but also known carriers are suitable.

Formulations suitable for parenteral administration include sterile aqueous solutions of the active ingredient 60, which solutions are preferably isotonic with the blood of the recipient. Such a composition may be prepared by dissolving the solid active ingredient in water to form an aqueous solution, sterilizing the solution and rendering it isotonic with the blood of the patient.

The formulations are stored in unit or multi-dose containers, such as sealed ampoules or vials.

In formulations suitable for nasal administration, the carrier is a coarse powder having a particle size of 20 to 500 microns, which is to be aspirated through the nose, for example by rapid inhalation through the nasal passage, keeping the vial containing the powder close to the nose.

In addition to the ingredients listed above, the compositions may also contain one or more ingredients, such as diluents, buffers, flavorings, binders, surfactants, thickeners, lubricants, preservatives (including antioxidants), and the like.

When formulated in a unit dosage form for either human or veterinary use, each unit dose will contain from 0.125 mg to 2 g, preferably from 1.25 pg-200 mg and optionally 12.5 Mg to 20 mg of active ingredient (all weight of peptide). base).

Further details of the invention are illustrated by the following non-limiting examples.

The following abbreviations are used:

HOBT 1-hydroxybenzotriazole

DCCI dicyclohexylcarbodiimide

DCU dicyclohexylurea

NMM N-methylmorpholine

DMF dimethylformamide

Pr isopropanol

Pr ₂ 0 diisopropyl ether pe petroleum ether

EtOAc ethyl acetate

Z is benzyloxycarbonyl

Bu tertiary butyl

BOC is a tertiary butyloxycarbonyl group

Bzl is benzyl.

The peptides were chromatographed on Merck silica gel plates using the following solvent system:

1. Methyl-free

2. n-butanol: acetic acid: water (3: 1: 1)

3. chloroform: methanol: 32% acetic acid (12: 9: 4)

4. chloroform: methanol: 880 ammonia (12: 9: 4)

5. ethyl acetate: n-butanol: acetic acid: water (1: 1: 1: 1)

6. chloroform methanol (8: 1).

Unless otherwise noted, amino acids have the L-configuration. Optical rotation was determined on a Bendix NPL automatic polarimeter.

The amino acid composition of the peptide hydrolyzates was determined using a Beckman-Spinco Model 120 ° C amino acid analyzer (6N hydrochloric acid evacuated at 110 ° C for 24 hours in sealed test tubes) or a Chromostak amino acid analyzer.

In general, the following methods were used to synthesize peptides:

a) The coupling was performed in DMF and DCCI was used as the coupling agent.

b) The amino acid ester hydrochlorides are converted to the free esters by addition of a tertiary base such as triethylamine or N-methylmorpholine.

c) HOBT is added at the coupling step if a partially peptide containing an optically active carboxyl-terminal amino acid is coupled to BOC.Tyr.D-Ala.Gly.Phe.OH in the partial condensation.

d) Switching lasts for 24 hours in a cold room at 4 ° C.

e) After coupling, purification is carried out by washing the mixture with acid and base to remove unreacted portions.

f) Alkaline saponification in aqueous methanol was carried out with an autotitrator at pH 11.5-12.0 with n sodium hydroxide.

g) The benzyloxycarbonyl protecting groups are removed by hydrogenolysis with a mixture of methanol and acetic acid in the presence of 10% palladium on carbon.

h) converting the acetate salts obtained by the above hydrogenolysis to the hydrochlorides by addition of the appropriate methanolic hydrochloric acid.

i) removing the benzyl protecting groups by hydrogenolysis in methanol in the presence of 10% palladium on activated carbon.

j) The tertiary butyl and tertiary butyloxycarbonyl protecting groups are removed in the presence of n hydrochloric acid acetic acid cleaner. The cleavage takes 60-90 minutes.

k) The OBu protecting groups protecting the alcoholic groups of threonine and serine are removed with trifluoroacetic acid containing 10% water for 90 minutes.

l) The final peptides are isolated as their hydrochloride and lyophilized from their aqueous solution.

1. Reference preparation

BOC.Tyr.Gly.Gly.Phe.OH

It is prepared according to the scheme depicted in Table 1, wherein the various protecting groups have the following meanings:

Z: benzyloxycarbonyl

Me: methyl

BOC: t-butyloxycarbonyl.

All coupling was carried out using dicyclohexylcarbodiimide in dimethylformamide and the reaction mixture was stirred at 4 ° C for at least 24 hours. The peptide methyl esters are saponified by the addition of 2N aqueous sodium hydroxide at pH 11.5 (measured in pH) in a 3: 1 solution of methanol and water (1). The protecting group Z is cleaved from the protected tripeptide (3) by hydrolysis with methanol in the presence of 10% palladium on activated carbon.

Table 2 gives the typical data, where Rf refers to thin layer chromatography on Merck silica gel and the indicated solvent systems.

184 704

Table 1

Tyr Gly Gly Phe Production Z.Gly.OH H Gly.OMe 96.4% Z.Gly (1) Gly.OMe Z.Gly (5) Gly.OH H.Phe.OMe 83.8% Z.Gly (3) - - Gly Phe.OMe 93.7% BOC.Tyr.OH H.Gly (4) - Gly Phe.OMe 79.2% BOC.Tyr Gly (5) - Gly Phe.OMe 77.6% BOC.Tyr Gly - Gly Phe.OH 86%

total production 40%

2. Reference preparation

B0C.Tyr.Gly.Gly.Phe.0H

Compound (4) was prepared as described in Example 1 and coupled with fully protected tyrosine (Bzl = benzyl) (see Table 3) to afford the protected tetrapeptide 20 (9). This product is listed in Figure 1.

The compound Bzl is cleaved by hydrogenolysis in methanol in the presence of a 10% palladium catalyst.

Table 2

Rf * * Mp. ° C crystallization solvent * Analysis 0) 0.65 ^a '0.59 ^a 66-67 EtAc / p.e. Cj 3 Hj gNj Oj Calculated%: C; 55.7, H: 5.7, N: 10.0 Found: C: 55.8, H: 5.9, N: 10.3 (2) 180 aqueous EtOH Ci ₂ H ₁₄ N ₂ 0j Calculated: C, 54.1; H, 5.3; N, 10.5 Found: C, 54.3; H, 5.6; N, 10.4 (3) 0.40 ° ⁵⁸ ethyl acetate / diisopropyl- ester C ₂ 3 Hj 5 N3 O5 Calculated: C, 61.8; H, 5.9; N, 9.8 Found: C, 61.6; H, 5.8; N, 9.6. (4) (H.C1) 0.77 ^c 182.3 MeOH / Calculated for C ₁₄ H 20 O ₁ C _{3 N 3} O ₄ : C, 51.0; H, 6.1; N, 12.8 k isopropanol Found: C, 51.3; H, 6.3; N, 12.6 (5) 0.33 ° (6) 0.8 ^a

Solvent (a) n-butanol, acetic acid, water (3: 1: 1) (b) methylethyl ether (c) chloroform, methanol, 32% acetic acid (12: 9: 4) * ppm: petroleum ether; EtAc: ethyl acetate; EtOH: ethanol 'V

184 704

Table 3

Tyr Gly Gly Phe Production OBZ1 BOC.Tyr .OH Η-Gly W Gly - -Phe.OMe OBzl BOC.Tyr - -Gly (9) Gly Phe.OMe 76.9% . OBzl BOC.Tyr BOC.Tyr- -Gly (10) Gly --Gly (6) Gly Phe.OH 86.2% Phe.OH 90% total production: 34.8% Rf * M.p. (° C) and solvent for crystallization Analysis (9) 0.83 ^a 0.34 ^b 0.70 ^a 138.2 C ₂₄ H ₄₀ N ₄ SHE,: 0.06 ^b EtAc calculated% found%: : C: 64.56, H: 6.33, N: 8.86 C: 64.42, H: 6.46, N: 8.55 ^ Solvent • (a) n-butanol, acetic acid, water (3: 1: 1)

(b) methylethylketone

Table 3 provides representative data for intermediates 9 and 10, where Rf refers to thin layer chromatography on Merck silica gel and the solvents mentioned above.

1-3. Example 40

H — Tyr.Gly.Gly.Phe.X ⁷ .OH

The peptides of the above formulas are prepared by condensing the protected tetrapeptide BOC.Tyr.Gly.Gly.Phe.OH (Reference formulas 1 and 2) with the corresponding carboxyl-protected H-X'-OR amino acid derivative, wherein - OR can mean -OMe,

OEt, OBu ⁴ , -OBzl, 0Bzl (p-NO ₂ ) etc. Depending on the method of removal of the protecting groups. -OMe and OEt reports In the case of gg, the carboxyl group can be liberated by alkaline saponification, in -OBu * by mild acidic acidolysis and in -OBzl or -OBzl (p-NO ₂ ) by hydrogenolysis.

Tyr Gly Gly Phe X ⁷

BOC.Tyr — Gly_L Gly — Phe.OH — HX ⁷ .OR

BOC.Tyr — Gly_2 Gly — Phe-X’.OR

H.Tyr -GlyGly- Phe-X ⁷ .0H

A) Preparation of BOC-Tyr.Gly.Gly.Phe.X .OR ⁷ - ⁷ wherein X = Gly, general method for the preparation of compound Ala, Val and R = Me,

A mixture of 0.92 mmol BOC.Tyr.Gly.Gly.Phe.OH, 0.92 mmol HCl.HX ^7.0 Me (e.g. glycine methyl ester hydrochloride) and 1.84 mmol HOBZ was dissolved in 5 mL DMF and 92 mmol NMM was added. The mixture was cooled to -10 ° C and DCM (0.92 mmol) was added. The solution was allowed to slowly warm to room temperature and stirred overnight. The DCU was filtered off, washed with a little DMF and the filtrate was concentrated in vacuo. The residue was dissolved in ethyl acetate and washed successively with 5% aqueous sodium bicarbonate, water, 5% citric acid and water. The solution was dried over anhydrous magnesium sulfate, evaporated and the residue was recrystallized from a suitable solvent.

184 704

Aj · <b '

Production % X ⁷ Op. (° C) (solvent for crystallization) R _f Analysis amino acid analysis 81 Gly 108-110 MeOH / EtOAc / pe 0.97 ³ 0.95 ⁴ 0.85 ^s + 1.37 ° (CRI), 5, methanol) C ₃₀ H ₃₉ N ₅ O ₉ .H ₂ O: Gly (3.08), calculated% C: 57.05, Tyr (1.00), H: 6.50, N: 11.09 Phe (1 Found: C, 57.02; H, 6.62; N, 10.64 77 Ala 115-117 EtOAc 0.87 ³ 0.95 ″ 0.91 ^s -8.81 ° (c = 0.5. methanol) C ₃ H _4, N ₉ ^s O Calculated% C, 59.32 H, 6.58; N, 11.16% Found: C 59.19, Η, 7.06; N, 10.74, Gly (1.98), Ala (1.09), Tyr (1.00), Phe (1.03) 89 With 150-151 Pr (EtOAc) Pr ₂ O 0.97 ³ 0.95 ^s -6.08 ° (c = 0.5 methanol) C ₃ 3 3 N 5 O ₉ Calculated Η4 ^ ,: C, 60.44; H, 6.92; N, 10.68% Found: C 59.94, Η, 7.02; N, 10.37 Gly (1.97), Val (0.94), Tyr (1.00), Phe (0.999).

Step B: BOC.Tyr.Gly.Gly.Phe.X'.OH - where X ⁷ = General Method for Preparation of Gly, Ala, Val

The protected pentapeptide of the formula BOC.Tyr.Gly.Gly.Phe.X'.OMe is dissolved in 6 ml of methanol, 3 ml of water are added and the pH is maintained between 11.5 and 12.0 with addition of sodium hydroxide until no calculated amount of alkali is added. The solution was concentrated in vacuo to remove the methanol and dilute with water.

The insoluble material was filtered off, the solution was extracted with ethyl acetate to remove the remaining ester and the pH of the aqueous phase was adjusted to pH 3 by addition of 0.7M citric acid. The precipitated peptide was extracted with ethyl acetate. The extracts were washed with water, dried over anhydrous magnesium sulfate and concentrated in vacuo.

X ⁷ Rf Analysis

Gly amorphous precipitate From EtOAc petroleum 0.84 ³ 0.74 ⁴ 0.83 \ _{2 3} C ₉ H 7 N 5 O ₉ H ₂ 0 - Calculated%: C, 56.40; H, 6.32; N, 11.34% Found: C, 56.33; H, 6.41; N, 11.07 Ala amorphous precipitate from EtOAc with Pr ₂ O ' 0.80 ³ 0.66 ⁴ 0.87 ^s C ₃ 0 _{3 9} Η Ng O .h O ₂ Calculated ^ »: C, 57.05; H, 6.50; N, 11.09% Found: C 57.20, H 6.43, N : 10.67 With amorphous precipitate From EtOAc petroleum 0.88 ³ , 0.85 ⁴ 0.93 ^s C32 E4 $ 3 ^ 4 Calculated: C, 59.89; Η: 6.75 Found: C, 59.98; H, 6.50.

Step C: General procedure for the preparation of H.Tyr.Gly.Gly.Phe.X'.OH HCl - where 50 X '= Gly, Ala, Val

To 100 mg of BOC.Tyr.Gly.Gly.Phe.X'.OH pentapeptide was added 1.0 N hydrochloric acid in 5 ml hydrochloric acid and 1 ml anisole. After stirring for 90 minutes at room temperature, the solvents were removed in vacuo. The residue was triturated with dry ether to give an amorphous solid. The product was dissolved in water, filtered and lyophilized to give a white solid as the hydrochloride salt of the peptide.

184 704

X ^{7 R} f Analysis amino acid analysis Gly (HC1) salt amorphous (lyophilized) 0.82 ³ 0.63 ⁴ 0.60 ' * 27.9 ° (C = 0.18 MeOH) ₂ Calculated% C 3 H 9 N i Oj HCl.Hj O: C, 50.97; H, 5.91; N, 12.93% Found: C 51.09, H 6.19, N 12, 54 Gly (3.02), Tyr (1.0), Phe (0.99), below (HC1) salt amorphous (lyophilized) 0.50 ³ 0.58 ⁴ 0.67 ⁵ + 22.3 ° (C = 0.2 MeOH) Calcd for C14H31N5O7.Ha.H2O: C, 51.85; H, 6.12; N, 12.6% Found: C, 52.14, H, 6.20; N, 12.10 Gly (1.95), Ala (1.01), Tyr (1.00), Phe (0.98). With (HC1) salt amorphous (lyophilized) 0.80 ² 0.67 ⁴ 0.79 ⁵ * 31.7 ° (C = 0, l, MeOH) Calculated for C %jHjjNjOj HaHHOH: C, 52.80; Gly (2.1), H: 6.52, N: 11.40, Val (1.02), Found: C, 52.89; Tyr (1.00), H, 6.45; N, 11.26. Phe (L, 05).

The following peptides were prepared by analogy to the preparation of the above examples and reference formulations according to methods accepted in peptide chemistry, with the following characteristic data. C-terminal derivatives are designated as follows:

-OMe: methyl ester

-NH ₂ : amide

-NHEt: ethylamide. ³⁰

In Example 10, BOC.Tyr.Gly.Gly.Tyr.Leu.OBu is dissolved in a few drops of boron trifluoride etherate in methanol / methylene chloride. An excess of an ethereal solution of diazomethane was added and the solution was allowed to stand at room temperature until the reaction with Pauly's reagent was negative. The solution is evaporated and the BOC and Bu protecting groups are removed from the residual solid by conventional means using a solution of acetic acid in hydrochloric acid in the presence of anisole.

The peptide hydrochloride was isolated in the foam of a colorless amorphous powder after lyophilization from an aqueous solution.

In Example 21, the peptide product was purified by chromatography on a silica gel (Merck) column and eluted with n-butanol: acetic acid: water = 3: 1: 1. Fractions positive to the Pauly's ⁽² Rf = 0.44) were combined, evaporated in vacuo and the residue was lyophilized from aqueous solution. The peptide base from Example 107 is prepared by reduction of H.Tyr.Gly.Gly.Phe.Leu.Ome, the reducing agent being lithium aluminum hydride or other known equivalent.

The peptide bases from Examples 113 and 114 were prepared by:

a) preparing the Z-leucinamide from a protected amino group;

b) converting the amide to nitrile (with phosphorus oxychloride),

c) reacting the nitrile with hydrogen azide to obtain protected leucintetrazole,

b) cleavage of the leucylamino group by hydrogenolysis in the presence of 10% palladium on charcoal,

e) collecting the pentapeptide in the usual manner.

Example number Compound, R _f (Methanol) 4th H.Tyr.D-Ala.Gly.Phe.Leu.OH HC1 0.63 ² , + 19.3 ° (c0.5) 5 (a). H.Tyr.D-Ala.Gly.Phe.MetOH HCl 0h2 ² , 0.60 ³ + 17.2 ° (c = 0.5) 5 (b). H.Tyr.D-Ala.Gly.Phe.Met.OMe 0.67 ² , 0.70 ³ ♦ 10.4 ° (c = 0.5) 6th H.Tyr.D-Ala.Ala.Phe.Leu.OH HC1 0.65 ² , 0.56 ⁴ + 1.68 ° (c = 0.5) 7th H.Tyr.Gly.Gly.Tyr.Leu.OH HQ. 0.47 ² , 0.50 ³ + 24.4 ° (c = O, 54) 8th HTHe.Gly.Gly.Tyr.Leu.OH HCL Me 0.55 ² , 0.57 ⁴ * 26.45 ° (c = 0.54) 9th H.Tyr.Gly.Gly.phe.Leu.OH HC1 Me Me 0.58 ² , 0.62 ⁴ * 21.77 ° (c = 0.5) 10th H.Tyr.Gly.Gly.lyr Le _U .OH 0.64 ² , 0.92 ³ * 21.0 ° (c = 0.51) 11th H.Tyr.Gly.D-Ala.Phe.Leu.OH HQ 0.60 ² , 0.90 ³ . 0.63 ⁴ 0.60 ² , -0,094 ³ 0.55 ⁴ * 36.67 ° (c = 1) 12th H.Tyr.Ala.D-Ala.Phe.Leu.OH HC1 * 6.43 ° (c = 1) 13th H.Tyr.Gly.Gly.Phe.Met.Thr.OH HQ 0.72 ^l , 0.65 ³ , 0.60 ⁴ , 0.60 ⁵ , ♦ 12.46 ° (c = O, 52) 14th H.Tyr.D.Ala.Gly.Phe.Met.Thr.OH HQ 040 ^l .0.49 ² , 0.68 ⁴

184 704

0.68 ² , 0.93 ³ , 0.93 ⁴ 0.68 ² , 0.74 ³ , 0.88 ⁴ 0.57 ² , 0.83 ⁴ 0.55 ¹ , 0.84 ³ .0.86 ⁴ η ss ² h «s ³ n

0.76 ^ 0.68 ^ 0.75 ³

0.57 ² , 0.86 ³

0.71 ^ 0.49 ^ 0.79 ³

0.51 ² , 0.85 ³ , 0.48 ⁴

O.76 ¹ , 0.85 ² , 0.65 ³

0.64 ² , 0.85 ³

0.66 ² , 0.93 ³

0.64 ² , 0.90 ⁴

0.62 ² , 0.86 ³ , 0.63 ⁴

0.50 ² , 0.51 ⁴

0.55 ', 0.89 ³ , 0.69 ⁴

0.63 ² , 0.92 ⁴

0.57 ² , 0.93 ³ , 0.82 ⁴

0.60 ² , 0.94 ³ , 0.85 ⁴

42nd

H, Arg.Tyr.Gly.Gly.Phe.Leu.OH 2HC1 0.38 ² , 0.61 ⁴ , H.Tyr.Ala.D.Ala.Phe.Met.Ome HCl 0.58 ² , 0.97 ³ , 0.88 ⁴

H.Tyr.Gla.D-Ala.Phe.Met.OMe HCl ^no ° ^{3 noe4} lI.Tyr.Ala.D-Ala.Plie.Met.OH HCl

H.Tyr.Gly.D-Ala.Phe.Met.OH HCI H.Arg.Tyr.Gly.Gly.Phe.Leu.Thr.OH 2HQ H.Tyr.D-Ala.GlyJPhe.Pro.OH HCI H. Tyr.D-Ala.D-Ala.Phe.Leu.OH HCI H.Tyr.D-Ala.Gly.Leu.Leu.OH HCI H.Tyr.De.Asn.Met.Leu.OH H.Tyr.Ala .Gly.Phe.Leu.OH HCi H.Tyr.Gly.Gly.Phe.Nle.OH HCI H.Tyr.D-Leu.GIy.Phe.Leu.OH HCI H.Tyr.Ala.Ala.Phe.Leu .OH HCI H.Tyr.Gly.Pro.Phe.Leu.OH HCI H.Tyr.D-Ala.Gly.D.Phe.Leu.OH HCI H.Tyr.Gly.Gly.Phe.D-Leu.OH HCI H.Tyr.Gly.Gly.D-Phe.Leu.OH HCI H.Tyr.Gly.Gly.Phe.Ue.OH HCI H.Tyr.D-Ala.Gly.C-Phenylglycyl-Leu.OHHQ H. Phe.D-Ala.Gly.Phe.Leu.OH HCI H.Tyr.D-Ala.Sar.Phe.Leu.OMe HCI H.Tyr.Gly.Ala.Phe.Leu.OH HCI H.Tyr.D- Ala.Gly.Phe.Thr.OH HCI H.Tyr.D-Ala.Asn.Phe.Leu.OH HCI H.Tyr.D.Ala.Gly.Phe.D-Leu.OMe HCI H.Tyr.D- Ala.Gly.Phe.D-Leu.OH HQ Ac

H.Tyr.D-Ala.Gly.Phe.Leu.OH HCl Ac

-Tyr- = 0 ⁴ '-acetyltyrosyl H.Tyr.D-Ala, Sar.Phe.Leu.OH HCl H.Tyr.D-Ala-Gly.Phe.Leu.Thr.OH HO H.MeTyr.Gly.Gly .Phe.Leu.OH HCI H.Tyr.D-Ala.Gly.Phe.Nle.OH HCI H.Tyr.Gly.Sar.Phe.Leu.OMe HCI H.Tyr.Gly.Sar.Phe.Leu.OMe HQ HD-Tyr.D-Ala.Gly.Phe.Leu.OH HCI HD-Tyr.D-Ala.Gly.Phe.D-Leu.OH HCI H.Tyr.Gly.Sar.Phe.Leu.OH HCI H .Tyr.D-Ala.GlyJ'he.D-Leu.Thr.OH HCI H.Tyr.D-Ala.Gly.Phe.D.Met.OH HCI H.Tyr.Pro.Gly.Phe.Leu.OH HQ. H.Tyr.D-Ala.Gly.Phe.D-Met.Thr.OH HCI H.Tyr.Sar.GlyJPhe.Leu.OH HCI H.Me.Tyr.D-Ala.GIy.Phe.Leu.OH H .Tyr.gly.D-Pro.Phe.Leu.OH HCl H.Tyr.D-Ala.D-Pro.Phe.Leu.OH Sodium 0.74% 0.81 ^ 0.74 ²¹ .

0.58 ² , 0.85 ⁴ 0.55 ² , 0.93 ³ , 0.72 ⁴ 0.57 ² , 0.64 ³ , 0.64 ⁴ 0.52 ² , 0.68 ⁴ , 0.56 ² , 0.7Ó ³ , 0.90 ⁴ 0.56 ² , 0.70 ³ , 0.90 ⁴ 0.55 ² , 0.68? ·

0.57 ² , 0.62 ³ , 0.50 ⁴ 0.59 ² , 0.80 ³ , 0.68 ⁴ 0.51 ² , 0.66 ³ , 0.55 ⁴ O.55 ² , 0.67 ³ , 0.58 ⁴ , 0.82® 0.92 ³ , 0.76 ⁴ , 0.66® 0.61 ² , 0, 76 ³ . 0.72 ³ , 0.91 ⁴ , 0.54 ² 0.90 ⁴ , 0.85 ³ , 0.52 ² 0.89 ⁴ , 0.82 ³ , 0.59 ² * 17.7 °

-3.66 '+30.19' + 4.22 ° +34.19 '+ 1.63 °

-1,52 '+43.71'

-4.43 °

13.5 '

-1.35 ° * 24.44 '+ 17.3 °

-33.6 °

-30.0 ° * 27.2 ° + 28.7 ° + 24.5 ° + 22.7 ° * 43.7 ° + 21.2 ° ♦ 3.69 °

-12.07 (c4), 52), (c = 0.2) (c = 0.2) (c = l) '(c = lj (c = 0.5), (c = 0.51) (c-1) (c = O, 5) (c = 0.2), (c = 1) (c = 0.4) (c = 0.2) (c = 0.6) (c = 0.5) ) (c = 0.5) (c = 0.4) (c = 0.52) (c = 0.6) (c = 0.48) (c = 0.52), (c = 0.4) ) (c-1) + 19.8 ° (c = 0.51) + 6.26 ° (c = 0.49) ♦ 39.6 ° (c = 0.5l) ♦ 31.5 ° (c = O, 53) 44.6 '(c = 0.5) + 9.89 ° (c = 1) * 5.13 ° (c = 0.5) + 25.5 ° (c = 0.5) * 23.2 '(c = 1.0) + 10-2 * 10.2 ° (c = 0.4) -20.7 ° (c = 0.5) -4.51 ° (c = 0.5) * 17.1 ° (c = 1.0) * 51.5 ° (c = O, 52) + 31.0 ° (¢ = 0.51) -5.6 ° (c = O, 2) * 47, 5 ° (c = 0.51) * 20.5 ° (c = 0.1) * 55.0 ° (c = 1.0) * 43.2 ° (c = 1.0) * 18.1 ° (c = 0.4) * 61.3 ° * 49.9 ° * 11.6 ° * 37.8 °

-39.0 '* 40.9 ° * 54.7 ° * 13.8 ° * 17 ° * 48.9 °

- = 42.7 ° * 9.03 '* 46.5 ° * 50.0 ° (c = 0.39) (¢ = 0.45) (c = 0.44) (c = O, 38)' (c = 1.0) ) (c = l) (c = l) (c = O, 4) (¢ = 1.0) (¢ = 0.5) (¢ = 0.1) (c = O.2) (¢ = 0.5) ) (c = l)

0.98 ⁴ , 0.99 ³ , 0.48 ² 0.91 ⁴ , 0.89 ³ , 0.63 ² 0.91 ⁴ , 0.85 ³ , 0.60 ² 0.68 ⁴ , 0.72 ³ , 0.62 ² 0.95 ⁴ , 0.95 ³ , 0.63 ² 0.94 ⁴ , 0.93 ³ , 0.57 ² 0.58 ²

0.55 ² , 0.81 ³ , 0.82 ⁴ 0.52 ² , 0.90 ³ , 0.92 ⁴ 0.49 ² , 0.85 ³

H.Metyr.Gly.Gly.Phe.Leu.OMe HQ H.MeTyr.D-Ala.Gly.Phe.Leu.OMe HCI H.Tyr.D-MeAla.Gly, Phe.D-Leu.OMe HCI H. Tyr.D-MeAla.gly.Phe.D-Leu.OH HCI H.Tyr.D-Ala.Gly.Phe.Leu.OMe HCI H.Tyr.D-Ala.Gly.Phe.D-Met.OMe HCI MD-Tyr.Gly.Gly.Phe.Leu.OH HCI H.MeTyr.D-Ala.GIy.Phe.D-Leu.OH HCI H.MeTyr.D-Ala.GlyJ'he.D-Leu.OMe HCI H.DOPA.Gly.Gly.Phe.Leu.OH HCl DOPA = 3,4-Dihydroxyphenylalanyl H.Tyr.MeAla.Gly.Phe.D-Leu.OH HCl H.Tyr.D-Ala.Gly.Phe.D -Leu.D-Thr.OH HCI 0.50 ² HD-Tyr.Gly.Gly.Phe.Leu.OMe HCI 0.63 ² , 0.97 ³ , 0.93 ⁴ H.Tyr.MeAla.Gly.Phe.D-Leu.OMe HCI 0.59 ² H.Tyr.D-Ala.GIy.Phe.D-Leu.NHEt HQ 0.61 ² , 0.93 ⁴ , 0.61 ^s H.MeTyr.D-Ala.Giv.Phe.D-Leu.NHEt HCl 0.58 ² , 0.92 ³ .0.96 ⁴

0.51 ² , 0.88 ³ , 0.79 ⁴

-101

184 704

75th Me-MeTyr.D-AIa.GIy.Phe.Leu.OH acetate 0.54 ² , 0.62 ³ , 0.66 * ♦ 39.2 ° (C = 0.4) 76th Me-MeTyr.Gly.Gly.Tyr.Leu.oh acetate 0.35 ?, 0.63 ³ , 0.87 * ♦ 31.3 ° (C = 0.4) 77th Me-MeTyr.Gly.Ala.Phe.Leu.OH HCl O.55 ² , 0.66 ³ , 0.80 * ♦ 38.2 ° (C = 0) 78th Me-MeTyr.Gly.Gly.Phe.Leu.OH HCl 0.45 ² , 0.65 ³ , 0.58 * ♦ 52.4 ° (C = 0) 79th Me-MeTyr.D-Ala.Gly.Phe.Leu.OMe HC1 0.45 ² , Ö.97 ³ , 0.97 * 80th Me-MeTyr.Gly.Gly.Phe.Leu.OMe HC1 0.38 ² , 0.92 ³ , 0.96 * 81st Me-MeTyr.D.Ala.Gly.Phe.D-Leu.OH HCl 0.40 ² , 0.86 ³ , 0.78 * ♦ 62.4 ° (C = 0, l) 82nd Me-MeTyr.D-Ala.Gly.Phe.D-Leu.OMe HCl 0.41 ² , 0.75 ³ , 0.96 * + 66.4 ° (C - 1.0) 83rd Me-D-MeTyr.GIy.Gly.Phe.Leu.OH HCl 0.36 ² , 0.89 ³ , 0.60 * -67.4 ° (C = 0.5) 84th Me-MeTyr.D-MeAla.Gty.Phe.D-Leu.OH HCl 0.33 ² , 0.76 ³ , 0.81 ' ♦ 58.4 ° _o (C = 0.2) 85th H.Tyr.Gly.Gly.Phe.Leu.Tyr.Gly.OH HCI 0.86 ³ , 0.84 ⁴ -4.79 (C = O, n 86th H.Tyr.D-Ala.GIy.His.Leu.OH 2HC1 0.26 ² , 0.47 ³ .0.52 * ♦ 20.8 ° (C = 0) 87th H.Tyr.D-AJa.Gly.Phe.D-Leu.Phe.Gly.OH HCl 0.63 ² ♦ 13.4 (C = 0.5) 88th H.Tyr.Gly.Gly.Phe.Met (O) .OH HCl 0.18 ² , 0.56 ³ , 0.51 ♦ 18.7 ° (C = 0.2) 89th H.Tyr.D-Ala.Gly.Phe.D-Leu.Lys.Lys.OH 0.44 ³ , 0.38 * +12.0 (c = 0.5) diacetate 90th H.Tyr.D-Trp.Gly.Phe.Leu.OH Sodium salt Ac 0.76 ² , 0.84 ³ -4.72 ° (C = 0) 91st H.Tyr.D-Ala.Gly.Phe.D-Leu.Tyr.OH 0.72 ² , 0.67 ^s ♦ 35.6 ° (C = 0.5) 92nd H.Tyr.D-Trp.Gly.Phe.D-Leu.OH HCl 0.76 ² ♦ 16.58 ° (C = 2) " 95th H.Tyr.D-Ala.Gly.Phe (4Cl) .D-Leu.OH HCl Me 0.42 * A -4.56 ° (C = 0.2) 96th H.Tyr.Gly.Gly.Tyr.Leu.OH HCI 0.55 ² , 0.93 ³ , 0.79 * ♦ 28.4 ° (¢ = 0.1) 97th H.Tyr.D-Ala.Gly.Phe (4Ci) .D-Leu.OMe HCl 0.65 ⁵ ♦ 36.5 ° (C = 0.5) 98th H.Tyr.GIy.Gly.Phe.D-Leu.OMe HCI 0.52 ^sec + 40.0 ° (¢ = 0.5) 99th H.MeTyr.D-Ala.Gly.Phe.D-Leu.NH ₂ HCl 0.56 ² ♦ 50.7 ° (C = l) 100th II.Tyr.Gly.Gly.Phe.D-Met.OMe HCI 0.62 ³ , 0.64 ^s ♦ 43.7 ° (¢ = 0.5) One hundred and first H.Tyr.Gly.Gly.Phe.D-Met.OH HCl 0.55 ² , 0.5 l ^s ♦ 38.3 (¢ = 0.5) 102nd H.MeTyr.D-Ala.Gly.Phe.D-Met.OMe HCI 0.62 ² , 0.75 ³ , 0.92 * ♦ 59.4 (¢ = 0.12) 103rd H.Tyr.Asn.Gly.Phe.D-Leu.OH 0.52 ² , 0.53 ³ A -11.8 ° (C = 0.52) 104th H.Tyr.Asn.Gly.Phe.D-Leu.OH 0.48 ² , 0.58 ³ A -21.0 ° (¢ = 0.5) 105th Me-MeTyr.MeAla.Gly.Phe.D-Leu.OH HCl 0.43 ² , 0.48 ³ ♦ 13.8 ° (¢ = 0.1) 107th H.Tyr.Gly.Gly.Phe.O-Acetilleucinol HCl 0.81 ³ , 0.84 *, 0.68 ⁵

-O-acetylucinol = -NH.CH (CH ₂ , CH (CH ₃ ) ₂ ).

, CH ₂ O.CO.CH ₃ 108th H.Tyr.GIy.Gly.Phe .Ieucinol HCI 0.81 ³ , 0.75 *, 0.68 ⁵ -ieucinol = -NH.CH (CH ₁ , CH (CH ₃ ) ₂ ) .CH ₂ .OH 109th H.Tyr.Ala («Me) Gly.Phe.D-Leu.OH HCl -Ala (aMe> = -NH.C (CH ₃ ) ₂ .CO- 0.55, 0.77, 0.59 ♦ 25.4 ° (C = O 52) 110th H.Tyr.Ala (aMe) .Gly.Phe.D-Leu.OMe HCI 0.20 ^, 0 ^, 0.95 ⁴ ♦ 28.5 ° (¢ = 0.5) 111th Hj3HomoTyr.Gly.Gly.Phe.Leu.OH HCl 0.53 ² , 0.63 ³ , 0.52 * -3.14 ° (¢ = 0.5) 112th H.Tyr.D-Ala.Gly .Phe.D-ŰHomoLeu.OH HCl 0.58 ¹ , 0.82 ³ , 0.49 ' ♦ 10.7 ° (¢ = 0.5) 113th H.Tyr.D-Ala.Gly.Phe.Leu-tetrazole HCl 0.60 ² , Ö.92 ³ , 0.96 ⁴ ♦ 48.2 ° (¢ = 0.1) 114th H.Tyr.D-Ala.Gly.Phe.D-Leu-tetrazole HCJ 0.60 ² , 0.90 ³ , 0.94 * ♦ 39.7 (¢ = 0.1)

CH (CH ₃ ) ₂ , tH ₂

-Leu-tetrazole = -NH-OH-C-NH ii \

Ν N

V

115th H.DOPA.D-Ala.Gly.Phe.D-Leu.OH HCl 0.56 ² , 0.72 ³ ♦ 29.9 ° (¢ = 0.1) 116th H.D-DOPA.D-Ala.GIy.Phe.D-Lcu.OH HCl 0.58 ² , 0.68 ³ ♦ 9.8 ° (¢ = 0.1) 119th H.MeTyr.D-Ala.Gly.Phe.D-Met.NHEt HCl 0.59 ² , 0.84 ³ , 0.90 * ♦ 59.7 ° (C = l) 120th H.MeTyr.D-Ala.Gly.Phe.D-Met.OH HCl 0.52 ² , 0.71 ³ , 0.69 * ♦ 34.7 ° (C = l) 121st H.MeTyr.D-Ala.Gly.Phe.D-Met.NH, HCl 0.52 ² , 0.72 ³ , 0.85 * ♦ 48.9 , (c = l) 122nd H.Phe (4Cl) .D-acetate 0.65 Leu.OH ^{Ala.Gly.Phe.D-3,} 00:45 * A - 0.28 ' (¢ = 0.2) 123rd H.Tyr.D-Ala.Gly.Phe (4CI) .D-Leu.NHEt HCl 0.63 ² , 0.68 ³ A, 0.68 ^s ♦ 40.7 ° (¢ = 0.5) 124th yiv H.Tyr.Gly.Gly.Tyr. Leu.OMe HCl 0.62 ² , 0.96 ³ , 0.15 ⁶ ♦ 18.9 ° (C = l)

^ A = chloroform: methanol: 321 acetic acid (120: 90: 5) ⁴ A = chloroform methanol: 880 ammonia (120: 90: 5)

-11Preparation of pharmaceutical preparations Preparation of tablets (20 mg tablets)

1 is a compound of the formula 20 mg e.g. M-Tyr-Gly-Gly-Phe-Me-Thr-OH HCl Lactose 76 mg Corn starch 10 mg gelatine 2 mg Magnesium stearate 2 mg

The compound of formula I above is admixed with lactose and corn starch. Granulated together with water-dissolved gelatin. The granules are dried, magnesium stearate is added and compressed to give tablets containing 110 of the active ingredient.

(5 mg / product) Cone

250 mg of compound of formula I, e.g. M-Tyr-D-Ala-Gly-Phe-Leu-OH HCl

Cone base (Other than Esterinum C) up to 100g

The suppository base was melted at 40 ° C. The compound of formula I is gradually converted to a white powder and stirred until homogeneous. Pour into suitable molds, 2 g per mold and allow to harden.

Other Esterinum C is a commercially available blend of saturated vegetable fatty acids with their mono-, di-and triglycerides. Launched by Henkel International, Düsseldorf.

Preparation of pessary (5 mg / product)

Compound of Formula I 5 mg pl H-Tyr-D-Ala-Gly-Phe-Met-OH HCl Lactose 400 mg

Povidon (polyvinylpyrrolidone) 5 mg

Magnesium stearate 5 mg

The compound of formula I above and lactose are mixed. Granulate with a solution of Povidone in 50% ethanol. The granules are dried, added with magnesium stearate, and pressed into suitable shaped punches to give 415 mg of active ingredient per pessary.

Preparation of lyophilized injection (100 mg / ampoule)

100 mg of compound of formula I, e.g. H-Tyr.D-Ala-Ala-Phe-Leu.OH HCl Water for Injections up to 2 ml

The compound of formula I above is dissolved in water for injection. The solution is sterilized by passing through a 0.2 μηι pore size membrane filter and collecting the filtrate in a sterile container. Aseptically, they are filled into sterile glass vials and lyophilized. Seal the ampoules with sterile aluminum seals with rubber stoppers.

The injection is reconstituted with water for injection or a sterile saline solution before use.

The weight of the compound of Formula I was always referenced to the peptide base.

Claims (6)

PATENT CLAIMS A process for preparing peptides or salts thereof, C 1 -C 4 alkyl esters, amides, N- (C 1 -C 4) alkylamides or acid addition salts thereof - wherein n is 0 or 1, X ⁵ is L-arginyl, X ³ is (D or L) -tyrosyl, (D or L) -N-methyltyrosyl, (D or L) -3,4-dihydroxyphenylalanyl, LO ^4'- methyltyrosyl, LO ⁴ '-acetyltyrosyl, L-phenylalanyl, L4-chlorophenylalanyl, or L- / 3-homothyrosyl, X ⁴ is glycyl, sarcosyl, α-methylalanyl, (D or L) -alanyl, (D or L) -N-methylalanyl, L-asparagmyl, L-isoleucyl, L-propyl- , D-leucyl or D-tryptophyl, X ³ is glycyl, sarcosyl, (D or L) -alanyl, prolyl or L-asparaginyl, X® is (D or L) -phenylalanyl, L-tyrosyl, LO ^4' -methyl-tyrosyl, L-4-chlorophenylalanyl, LC-phenylglycyl-L-leucyl, L-methionyl, or L- hisztidilcsoport, X ⁷ is glycyl, or (D or L) -leucyl, (D or L 1 -methionyl, L-alanyl, L-isoleucyl, L-norleucyl, L-valyl, L-methionyl sulfoxide, L-threonyl, L-prolyl or D-O-hinooleucyl, X * is (D or L) -threonyl, L-lysyl, D-phenylalanyl, or L-tyrosyl, X 'is glycyl or L-lysyl, R is hydrogen or C 1-4 alkyl, R ¹ is the hydroxy group of the 1-carboxyl group of the C-terminal amino acid or the group -CH ₂ OR ⁴ - which is a substituent of the 1-carboxyl group - wherein R ⁴ is hydrogen or C 1 -C 4 alkylanoyl or 5-tetrazolyl; q is 0 or 1, with the exception of compounds of formula V and RL-Phe-L-Ile-Gly-L-Leu-L-Met-OH and salts thereof, C1-C4 alkyl esters, amides, N1-4 alkylamides and acid addition salts thereof - wherein R is as defined above, X ⁷ is L-leucyl or L-methionyl and X ³ is either L-tyrosyl and then X ⁶ is L-phenylalanyl or X ³ is L -tirozilcsoport and X ⁶ is L-4-klórfenilalanilcsoport - preparation, characterized in that a) a compound of Formula ii wherein R is as defined in Formula I and Y * is an N-terminal radical fragment of Formula I with a compound of Formula III wherein Y ² is a radical fragment of a compound of Formula II additional C-terminal radical moiety - condensed and optionally compounds II and III are protected and / or activated and optionally deprotected, b) reacting the corresponding C 1 -C 4 alkyl ester of the compound of formula I with ammonia or monoalkylamine to produce amides of formula I or N- (C 1-4) alkyl amides, or c) esterifying a compound of formula I wherein R * is hydroxy of the 1-carboxyl group of the C-terminal amino acid to produce C 1-4 alkyl esters of formula I; or Λ 184 704 d) and then preparing the compounds of formula I R ¹ represents the C-terminal carboxyl group of the hydroxy-1 aminosavesoport saponifying the ester of formula I 5 the product into a salt or an acid addition salt or base form. Priority: November 23, 1976 2. A process for preparing peptides of formula IX or salts thereof, C 1 -C 4 alkyl esters, amides, N-C 1 -C 4 alkyl amides or acid addition salts thereof, wherein X ² is L-arginyl, n is 0 or 1, X 'is (D or L) -tyrosyl, (D or L) -N-methyltyrosyl (O or L) -3,4-dihydroxyphenylalanyl, L-O-methyltyrosyl, L-O ⁴ ' -acetyltyrosyl, or L-phenylalanyl, X ⁴ is glycyl, sarcosyl, (D or L) -alanyl, (D or L) -N-methyl-alanyl, -isoleuzyl, L-prolyl or D-leucyl, X ⁵ is glycyl, sarcosyl, (D or L) -alanyl, 20 (D- or L) -prolyl or L-asparaganyl, X ⁶ represents (D or L 1 -phenylalanyl, L-tyrosyl, L-O ⁴ -methyl-tyrosyl,? -L-phenylalanyl, L-tyrosyl, L-leucyl or L-methionyl, X ⁷ is glycyl, (L or D) -leucyl, (D or L) -methionyl-L-alanyl, L-isoleucyl, L-norleucyl, L-valyl, β-threonyl or L -prolilcsoport, X ⁸ is (D or L) -treonyl and p is 0 or 1, provided that when n and p are 0 and X ³ , X ⁴ , X ⁵ and X ^{6 are} L-tyrosyl, glycyl, glycyl and L-phenylalanyl, then compounds and salts thereof, C1-4 alkilésztcreit, amides of formula X than ⁷ L metioniltól and L-om leucyl group and excluding the H-Phe-L-Ile-Gly-L-Leu-L-Met-OH , N1-C4 alkyl amides and acid addition salts thereof, characterized in that gg a) a compound of Formula II wherein R is as defined in Formula I and Y ¹ is an N-terminal radical fragment of Formula I with a compound of Formula III wherein Y ² is a radical fragment of Formula II condensation of the additional C-terminal radical fragment to the compound of formula (II) - where appropriate, the compounds of formula (II) and (III) are protected and / or activated and, if desired, deprotected, b) reacting the corresponding C 1 -C 4 alkyl ester of the compound of formula I with ammonia or monoalkylamine to produce amides of formula I or C 1 -C 4 alkylamides, or c) The I C1 -C4 alkyl esters of formula R ¹ is the C-terminal _ _n aminosavesoport esterifying a compound of formula I with 1-hydroxy group of the carboxyl group d) saponification of the ester of formula gg I to produce compounds containing R ¹ in the formula I where R ¹ is the hydroxy group of the C-terminal amino acid group, and then converting the product to a base or a salt or an acid addition salt thereof. Priority: March 3, 1976 3. A process for preparing peptides of formula IX, or salts thereof, C 1-4 alkyl esters, amides, N- -alkylamides or acid addition salts thereof - wherein n is 0 or 1 and X ² is L-arginyl, X ³ is L-tyrosyl, LN-methyltyrosyl, L-3,4-dihydroxyphenylalanyl, L-O ⁴ -methyltyrosyl, LO ^4, -acetyltyrosyl, L-phenylalanyl, X ⁴ is glycyl, sarcosyl, (D or L) -alanyl, (D or L) -N-methyl-alanyl, L-isoleucyl, L-prolyl, or D-leyl, ^And X is glycyl, sarcosyl, (D or L) -alanilcsoport or -prolilcsoport (D or L); X® is L-phenylalanyl, L-tyrosyl, L-O ^4' -methyltyrosyl, or LC-phenylglycyl, X ⁷ is glycyl, L-leucyl, L-methionyl, L-alanyl, L-isoleucyl, L-norleucyl, L-valyl, or L-prolyl, X ⁸ is L-threonyl and p is 0 or 1, provided that n and p are both 0 and X ^3, X ^4, X ^s and X®jelentése L-tyrosyl, glycyl, L-phenylalanyl glicilés, X ⁷ other than L-methionyl or L-leucilcsoport - preparation, characterized in that a) a compound of Formula II wherein R is as defined in Formula I and Y ¹ is an N-terminal radical fragment of Formula I with a compound of Formula III wherein Y ² is a radical fragment of Formula II condensation of the additional C-terminal amino moiety to the compound and, where appropriate, the compounds of formula II and III are protected and / or activated and, if desired, deprotected, b) the amides of formula I or the N- (1-4C) alkyalainides of the compound of formula I. the corresponding C 1-4 alkyl ester of the compound are reacted with ammonia or monoalkylamine, or c) esterifying a compound containing the hydroxy group of the 1-carboxyl group of the C-terminal amino acid of the C 1 -C 4 alkyl esters of formula I, or d) saponifying the ester of formula I to produce compounds having the L-carboxyl group of the C-terminal amino acid group of formula (1) and converting the product into a base or a salt or an acid addition salt thereof. Priority: January 26, 1976 The process for the preparation of a pharmaceutical composition according to claim 1, wherein the peptide of formula I, wherein the substituents are as defined in claim 1, is in the form of a pharmaceutically acceptable carrier and / or an acid addition salt thereof. admixed with excipients and converted to a pharmaceutical composition. Priority: November 23, 1976 5. A process for the preparation of a pharmaceutical composition according to claim 2 wherein the peptide of formula I, wherein the substituents are as defined in claim 2, is in the form of a pharmaceutically acceptable carrier and / or carrier. -131 184 704 Icai is mixed and turned into a pharmaceutical composition. Priority: March 3, 1976 5 6. A process for the preparation of a pharmaceutical composition according to claim 3, wherein the peptide of formula I wherein the substituents are a'2. The pharmaceutically acceptable salt or acid addition salt thereof as defined in claim 1, which is a pharmaceutically acceptable salt or acid addition salt thereof, and is formulated into a pharmaceutical composition.