HU195234B - Process for production of nonapeptide and dekapeptide hormone-analogs freeing lutheining hormones with antagonizing hormone effect and medical preparatives containing them - Google Patents

Abstract

The present invention relates to a process for the preparation of the novel nona or decapeptide LHRH analogs of formula (I) and their pharmaceutically acceptable acid addition salts. In the formula (I), A, B and C independently of one another are optionally substituted natural amino acid residues, F are novel guanidino-substituted water-soluble, non-natural amino acid residues, and H is a natural amino acid residue or a substituted amino group. According to the present invention, a) a protected polypeptide corresponding to a compound of formula I optionally attached to a solid support is removed and optionally a covalently bonded solid support is removed; or b) linking two fragments corresponding to the compound of formula (I) in the appropriate order; and optionally converting a compound of formula (I) into a pharmaceutically acceptable acid addition salt; or ii) converting an acid addition salt of a compound of formula I into a pharmaceutically acceptable acid addition salt. Due to their LHRH antagonist activity, the compounds of formula (I) are useful as pharmaceutical active ingredients for a wide variety of uses, most preferably for the inhibition of ovulation and treatment of endometriosis in females, and in particular for the inhibition of spermatogenesis and treatment of prostate tumors in male animals. A — B — C-5er — T ^ r-F — Leu-Arg — Pro — H (I) 12345678 9 10 -1-

Description

The present invention relates to novel nonapeptide and decapeptide hormone analogs and their pharmaceutically acceptable acid addition salts which have a luteinizing hormone releasing hormone (LHRH) antagonizing activity.

Luteinising hormone (LH) and follicle stimulating hormone (FSH) are released from the anterior pituitary gland under the control of the releasing hormone (LHRH), the latter being produced in the hypothalamus. LH and FSH stimulate the synthesis of steroid hormones and gamete by acting on the gonads. The rapid release of LHRH, and consequently the release of LH and FSH, regulates the reproductive cycle in both mammalian animals and man.

LHRH also indirectly affects the placenta and gonads by causing the release of the hormone gonadotropin (hCG) in the middle fetus.

LHRH antagonists can be used to control fertility. Antagonists inhibit ovulation in females and spermatogenesis in males. In connection with these effects, the normal circulating levels of sex steroids from gonads are reduced and the weight of the male and female adrenal organs is also reduced. In pets, this effect promotes weight gain during fattening, in pregnant animals, promotes abortion and generally acts as a chemical sterilizer.

Natural LHRH is a decapeptide composed of natural amino acids (all amino acids are in the L-configuration except for the optically inactive glycine). The hormone amino acid sequence has the following sequence:

(pyro) Glu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH ₂ 1 234557 89 10

Many analogs of the above natural hormone have been studied, but most of them have not shown enough biological activity to be useful in medicine. However, some well-chosen modifications have resulted in increased biological efficacy. The most significant effect to date has been obtained when the glycine residue at position 6 is replaced by a D-amino acid residue.

In addition to LHRH agonist analogues, LHRH analogues which competitively antagonized LHRH were also prepared. The antagonist analogs are either lacking the 2-position histidine or are replaced by another (Vale, W. et al., Science 176, 933 (1972)). In general, it has been found that the highest efficiency is obtained when a D-amino acid is inserted at the above site in the sequence [Rees, R.W.A. et al., J. Med. Chem. 17, 1016 (1974)].

It has also been found that modification of position 6, which produces an agonist effect without modification of position 2, results in an increase in the antagonist effect of the position modified analogs [Beattie, C.W. et al., J. Med. Chem. 18, 1247 (1975); Peptides 1976, p. 427, Ed. 1'Universite de Bruxelles, Belgium (1976)].

Based on the above two major changes, which resulted in a series of potent LHRH antagonist analogs, a further enhancement of the antagonist effect could be achieved by further modifying the analogs already modified at positions 6 and 2 to 1,

3 and / or 10 [Coy, DH et al., Peptides 1976, p. 462 Ed. 2 de l'University de Bruxelles, Belgium, 1976; Rivier, JE et al, Life Sci 23, 869 ~ _Q. (1978); Dutta AS et al., Biochem. Biophys. Gap. Comm. 81, 382 (1978); Humphries J. et al., Biochem. Biophys. Gap. Comm. 85, 709 (1978)]. It has also been shown that N-acylation of the amino acid at position 1 is highly advantageous [Channabesavaia K. et al., Biochem. Biophys. Gap. Comm., 81, 382 (1978); Coy, DH et al., Peptides, Structure and Biological Function, page 775, Pierce Chemical Company (1979).

Also disclosed is (N-Ac-Dp-Cl-Phe ¹ , Dp-Cl-Phe ² , D-Trp ³ , D-Arg ⁶ , D-Ala 10) -LHRH [Coy DH Endocrinology 110, 1445 ( 1982)]. Alternatively, it has been found that the D-Ala ⁴ modification reduces the LHRH antagonists [Pedroza, E., Martinez JA, DH Coy, A. Arimura and ⁵ Schally AV, Int. J. had access. 23, 294 (1978)].

Because the antagonistic effect is due to the fact that these compounds compete with LHRH for their respective receptor sites, the above compounds must be administered in high doses to displace the natural peptide from the binding site. Particularly preferred in this regard are compounds which have a high potency and a long lasting effect. Also, slow release compounds from sustained release formulations are significant. The disadvantages of the currently known analogs are that they need to be administered in relatively high doses, which leads to the emergence of toxicity problems and other side effects.

The novel high potency nonapeptide and decapeptide _R -LHRH analogs of the present invention contain a 2-substituent, i.e., an antagonist.

-2195234 - and, to enhance the effect, the glycine residue at position 6 is also replaced by a new guanidino-substituted water-soluble amino acid residue that does not occur naturally. The effect of the analogs is further enhanced by substitutions at the 1-, 2-, 3-, 4-, 7- and / or 10-positions. The use of the compounds of the present invention and pharmaceutical compositions containing them as active ingredients are also disclosed.

More particularly, the present invention relates to a process for the preparation of novel nonapeptide and decapeptide LHRH analogs of formula (I) and their pharmaceutically acceptable acid addition salts. In the general formula (I)

A is N- (C 2 -C 5) alkanoyl-L-prolyl, N- (C 2 -C 5) alkanoyl-Dp-chlorophenylalanyl, N- (C 2 -C 5) alkanoyl-seryl, or N- (C 2 -C 5) alkanoyl-3- (2-naphthyl) -D-alanyl,

B is D-p-chlorophenylalanyl or D-p-fluorophenylalanyl,

C is D-tryptophanyl or 3- (2-naphthyl) -D-alanyl,

F is an amino acid residue derived from a compound of formula II wherein n is 3 or 4,

R 1 is C 1 -C 6 alkyl, R ₂ is hydrogen, C 1 -C 6 alkyl, or - (CH ₂ ) n - (R ₄ ) _{2 -} wherein n is 1, 2, 3, 4 or 5 and R ₄ are C 1 -C 4 alkyl, or

R 1 and R _{2 taken} together form a ring of formula (f)

H is D-alaninamide, glycinamide, or -NHR _{5 in} the latter formula

R ₅ is lower alkyl or -NHCO ₂ NH ₂ . Replacement of the L-histidyl moiety at position 2 of LHRH with one of the amino acid residues listed above for B is a prerequisite for the compound to exhibit LHRH antagonistic activity. Substituting the glycyl group at the 6-position of the LHRH with the substituents designated for F significantly increases the antagonist activity. With the other substituents at the 1-, 2-, 3-, 4-, 7- and 10-positions, the antagonist effect can be further enhanced.

As used herein, for convenience, the amino acids shortening proposed by abbreviations commonly used in peptide chemistry, IUPAC-IUB Commission on Biochemical Nomenclature [Biochemistry, 11, 1726 (1972)] ²⁵ used. The above abbreviations denote L-amino acids, with the exception of optically inactive glycine, unnatural amino acids, natural achiral amino acids, or other designated D-amino acids, and amino acids that are normally substituted in the LHRH. - occupy positions 2, 3, 4, 6, 7 and 10. The peptide sequences are also written according to the generally accepted designation,

3θ, i.e. the N-terminal amino acid on the left and the C-terminal amino acid on the right.

Other abbreviations used in this description are listed below.

Amino acid substitutes not found in nature are abbreviated as follows:

Aminosavnaradék Abbreviation 3- (2-naphthyl) -D-alanyl D-Nal (2) ρ-fluoro-D-phenylalanyl D-p-F-Phe p-chloro-D-phenylalanyl D-p-Cl-Phe N, Ν'-guanidino-dimethyl-D -homoarginil D-DMH N, N'-guanidino-diethyl-D -honoarginíl D-Deh Ν, N'-guanidino-dipropyl-D- -honoarginíl- D-Dph Ν, Ν'-guanidino-diisopropyl-D -honoarginíl D-Dihydroxy N, N'-guanidino-dihexyl-D- -honoarginíl- D DHH N-guanidino-isopropyl-D- -honoarginil D-IPH N-guanidino-propyl-D- -homoarginil D-PRH Ν, Ν'diizopropi1-guanidino-D- -argin.il D-CHROME DN ^£ -2-imidazolin-2-yl-lysyl D-Dhi Ν, N'-guan idino-dimethyl-arginine1 Dm a N-guanidino-ethyl N'guanidino-dimethyl-aminopropyl -honoarginíl WSH

-3195234

6

For further simplification, since the natural LHRH has the following sequence:

(pyro) Glu-His-Tfp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH ₂ 12 23 456789 10 nona and decapeptides in which the corresponding amino acid residue is replaced by another amino acid residue or other moiety is abbreviated by referring to the position and identity of the substitution back to the parent compound LHRH. *

For example, (pyro) Glu-DpF-Phe-Trp-Ser-Tyr-D-Dih-Leu-Arg-Pro-Gly-NH ₂

6 7 8 9 10, wherein the glycine at position 6 with the D-Dih residue, the histidine at position 2 with the nonapeptide DpF-PheN-Ac-Pro-DpF-Phe-Trp-Ser 1 2 3 4 [NAc-Pro ^1, the DPF-Phe ^z, D-Dihydroxy ^6, Pro NHEt] LHRH abbreviated.

As used herein, the term pharmaceutically acceptable acid addition salt refers to salts that have a biological activity consistent with that of the parent compound and do not cause undesirable toxic symptoms. The above salts include, for example, inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, or organic acids such as nitric, oxalic, tartaric, succinic, maleic, fumaric, acid addition salts with tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalene disulfonic acid, polygalacturonic acid.

The term lower alkyl as used herein refers to a straight or branched C 1 -C 4 saturated hydrocarbon chain such as methyl, ethyl, n-propyl, isopropyl, η-butyl, isobutyl, sec-butyl or tertiary. butyl. C 1-6 alkyl includes the same as the lower lower alkyl group, but also includes C 5 and C 6 chains such as n-pentyl, n-hexyl or other C 5-6 branched hydrocarbon chains. .

N-Ac stands for N-acetylated amino acid residue in accordance with generally accepted nomenclature.

Preference is given to compounds of formula (I) obtained by the process of the invention wherein:

A is N-Ac-L-Pro, N-Ac-D-Ser, N-Ac-D-p-Cl-Phe, N-Ac-D-Nal (2),

is replaced by (DpF-Phe ² , D-Dih ⁶ ) -LHRH or

Dihydroxy-Tyr-D-Leu-Arg-Pro-NHEt

6 7 8 9

B is D-p-F-Phe or D-p-Cl-Phe, C is D-Trp or D-Nal (2),

D is Ser,

E is Tyr,

3θ F is an amino acid residue derived from a compound of formula II wherein n is 3 or 4,

R 1 is a group -NHR ₃ and

R ₃ is methyl, ethyl, η-propyl, isopropyl or n-hexyl and

R ₂ is hydrogen or R ₃ ; or in which

R 1 and R ₂ together are 2-imidazolin-2-yl;

H is D-AlaNH ₂ , GlyNH ₂ or NHEt. In general, compounds having F in place of N, N'-guanidino-disubstituted-D-arginyl or -D-homoar45 ginyl are also preferred.

Even more preferred are the compounds of formula I wherein

A is N-Ac-L-Pro, N-Ac-D-Nal (2), or N-Ac-D-p-Cl-Phe,

B is D-p-F-Phe or D-p-Cl-Phe, C is D-Nal- (2) or D-Trp,

D is Ser,

E is Tyr,

F is D-Dmh, D-Deh, D-Dph, D-Dhh or D-Dhi and

H is D-AlaNH ₂ , GlyNH ₂ or NEt; and peptides having the following sequence:

N-Ac-L-Pro-DpF-Phe-D-Nal (2) -Ser-Tyr-F-Leu-Arg-Pro-GlyNHj, wherein F is D-Dmh, D-Deh, D-Dph, D-. Dhh, D-Dhi or D-Prh;

N-Ac-L-Pro-DpC 1 -Phe-D-Trp-Ser-Tyr-F-Leu-Arg-Pro-NHEt, where F is D-Dmh, ⁶⁵ D-Deh, D-Dph, D-Dhh , D-Dhi, or D-Prh;

-4195234

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-F-Leu-Arg-Pro-D-AlaNH ₂ , where F is D-Dmh, D-Deh, D- Dph, D-Dhh, D-Dhi or D-Prh;

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-F-Leu-Arg-Pro-GlyNH ₂ , wherein F is D-Dmh, D-Deh, D-Dph, D-Dhh, D-Dhi or D-Prh;

N-Ac-D-Nal (2) -DpF-Phe-D-Trp-Ser-F-Leu-Arg-Pro-GlyNH ₂ , wherein F is D-Dmh, D-Deh, D-Dph, D-Dhh , D-Dhi or D-Prh;

N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-F-Leu-Arg-Pro-GlyNH ₂ , wherein F is D-Dmh, D-Deh, D-Dph , D-Dhh or D-Dhi;

N-Ac-D-Nal (2) -Dp-Cl-Ph-D-3Pal-Ser-Tyr-D-Deh-Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-D-Nal (2) - Dp-Cl-Phe-D-Trp-Ser · -Tyr-F-Leu-Arg-Pro-D-AlaNH ₂ , wherein F is D-Dmh, D-Deh, D-Dph, D-Dhh or D-Dhi;

The most preferred compounds of formula I are as follows:

N-Ac-L-Pro-D-p-F-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-NHEt;

N-Ac-L-Pro-D-p-F-Phe-D-Trp-Ser-Tyr-D-Dph-Leu-Arg-Pro-NHEt;

N-Ac-L-Pro-D-p-F-Phe-D-Trp-Ser-Tyr-D-Leu-DML-Arg-Pro-NHEt;

N-Ac-D-Nal (2) -D-p-F-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-NHEt;

N-Ac-D-Nal (2) -D-p-Cl-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-NHEt;

N-Ac-D-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Dmh-Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-D-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Deh Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-D-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Dph-Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-D-Nal (2) -DpF-Phe-D-Trp-Ser · -Tyr-D-Dhh-Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-D-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Dhi-Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dmh-Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dph-Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dhh-Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dhi-Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-DT rp-Ser-Tyr-D-Dmh-Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-DT rp-Ser-Tyr-D-Deh-Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dph-Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dhh-Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dhi-Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-D-AlaNH ₂ ;

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-SerTyr-D-Dph-Leu-Arg-Pro-D-AlaNH ₂ ;

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dhh-Leu-Arg-Pro-D-AlaNH ₂ ;

N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-D-AlaNH ₂ ;

N-Ac-D-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-D-AlaNH ₂ ;

N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dmh-Leu-Arg-Pro-GlyNH ₂ ;

N-Ac-D-Nal (2) -D-p-Cl-Phe-D-Trp-Ser-Tyr-D-Dph-Leu-Arg-Pro-D-AlaNHj;

N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dhh-Leu-Arg-Pro-D-AlaNH ₂ ;

N-Ac-D-NaI (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dhi-Leu-Arg-Pro-D-AlaNH ₂ ;

Pharmaceutically acceptable salts of the compounds listed above may also be prepared by the process of the present invention.

The compounds of formula (I) according to the invention, and in particular their pharmaceutically acceptable acid addition salts, exhibit surprisingly potent and prolonged LHRH antagonistic activity.

The effect is primarily measured by measuring the inhibition of ovulation in rats, Corbin A. and Beattie C.W. [Endocrine Slit. Commun. 2 1 (1975)] or by measuring the inhibition of LH release and ovulation in rabbits, Phelps,

C. P. et al., Endocrinology 100, 1526 (1977).

In addition to measuring the inhibition of LHRH-induced FSH and LH in rats, in vivo, Vilchez-Martinez, J.A. et al., Endocrinology 96, 1130 (1975), or measuring the inhibition of LH and FSH release by dispersed cell culture from the anterior pituitary gland by radioimmunoassay, Vale W. et al., (Endocrinology 91: 562 (1972)). .

The compounds of the present invention are useful in the following fields because of their LHRH antagonist activity:

- female contraception - cessation or delay of ovulation - initiation of childbirth - synchronization of ovulation - cessation of sex cycle - increase of growth in females - luteolysis, induction of menstruation - initiation of early, first trimester miscarriage - treatment of endometriosis - cystitis Stein-Leventhal) - treatment of uterine cancer - treatment of benign prostatic hypertrophy and prostate cancer - male contraception

-5195234 — Treatment of diseases caused by excessive gonadal hormone production, elimination of premenstrual bleeding in dogs in both sexes, elimination of menopausal symptoms.

The above-listed compounds of the present invention are particularly useful in inhibiting ovulation and in treating endometriosis in female subjects, whereas in male subjects, they are particularly useful in inhibiting spermatogenesis and treating prostate tumors.

The compounds of the present invention may be administered to the subject to be treated, if necessary or desired, in the form of a pharmaceutical composition, in an amount sufficient to produce the desired effect. Pharmaceutical compositions containing the compounds as active ingredients may be administered to the subject to be treated by any known route of administration, i.e., oral, parenteral, including subcutaneous, intramuscular and intravenous administration, vaginally (particularly for contraception), rectally, including sublingual administration. ), may also be administered transdermally or intranasally.

The most preferred form of administration will be selected according to the purpose of the treatment, the particular active ingredient, the subject being treated and the opinion of the attending physician. The active compounds may also be used in the form of slow release, sustained release or implant formulations, as described in more detail below.

Generally, the active ingredient will be administered in a dosage of 0.01 to 10 mg / kg body weight / day as listed above for therapeutic purposes. More preferably, the dosage will vary between 0.1 and 5.0 mg / kg body weight / day. The drug may be administered in a single daily dose, or in multiple portions, or in a slow release form, for maximum effect.

The precise dosage and dosage will, of course, be dependent on the individual requirement of the subject being treated, the mode of treatment, the severity of the disease or the degree of need, and of course the judgment of the attending physician. Generally, parenteral administration will achieve the desired effect at a lower dose than with other modes of administration that are more dependent on absorption.

The present invention also relates to the preparation of pharmaceutical compositions containing the compounds of formula (I) or pharmaceutically acceptable salts thereof as active ingredients. The pharmaceutical compositions are prepared by admixing the compounds of formula (I) or salts thereof according to the invention with a pharmaceutically acceptable non-toxic carrier. Formulations for parenteral (subcutaneous, intramuscular, or intravenous) administration may be in the form of liquid solutions or suspensions, and vaginal or rectal formulations are preferably in a semi-solid form such as creams or suppositories. Oral and oral formulations may be in the form of tablets or capsules, while intranasal formulations may be in the form of powders, nasal drops or aerosols.

The compositions are preferably administered in unit dosage form and prepared according to well-known pharmaceutical formulations such as those described in Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa. (1970). Formulations for parenteral administration generally contain sterile water or saline, polyalkylene glycols such as polyethylene glycol, vegetable oils, saturated naphthalene derivatives or the like as carriers. Formulations for vaginal or rectal administration, for example suppositories, may contain, for example, polyalkylene glycols, petroleum jelly, cocoa butter or the like as carriers. Formulations for inhalation may contain, for example, lactose as a carrier, whereas liquid, aqueous or oily solutions may take the form of nasal drops. For oral use, carriers which may be used are generally sugars, calcium stearate, magnesium stearate, gelatinized starch or the like.

It is particularly desirable to provide sustained release formulations of the compounds of the present invention that are sufficient to be administered once a week or even once a year to achieve the desired effect. The dosage form for use above includes, for example, a nontoxic pharmaceutically acceptable acid addition salt of the active ingredient which is poorly soluble in body fluids, such as a polybasic acid, sulfuric acid, citric acid, tartaric acid, tannic acid, acid addition salts with disulphonic acid, polygalactruronic acid or the like.

In addition, the compound of formula (I), or preferably a relatively insoluble salt thereof, as described above, may be formulated in a gel such as an aluminum monostearate gel such as sesame oil to provide an injectable preparation. Especially preferred are, for example, zinc salts, zinc tannate salts or pamoate salts. In another embodiment of an injectable, sustained release sustained release formulation, the active ingredient or salt is dispersed in a non-toxic, non-toxic, non-antigenic polymer, such as polylactic acid / polyglycolic acid described in U.S. Patent 3,773,919. ab-be-6195234 closed. The compounds of formula (I), or more preferably their relatively insoluble salts thereof, may also be formulated as cholesterol-based pellets, particularly for veterinary use. Other slow release, sustained release, and implantable formulations, such as liposomes, are described in [Sustained and Controlled Release Drug Delivery System, J.R. Robinson ed., Marcel Dekker, Inc., New York, 171978]. Further references to LHRH-type compounds can be found, for example, in U.S. Patent No. 4,010,125.

The novel polypeptides of formula (I) may be prepared by methods commonly used in peptide chemistry. An excellent summary of the various procedures can be found in [J.M. Stewart and J.D. Young: Solid Phase Peptide Synthesis, W.H. Freeman Co., San Francisco, 1969; and J. Meienhofer; Hormonal Proteins and Peptides, Vol 2, page 46, Academic Press (New York), 1973], for solid phase synthesis, while [E, Schroder and K. Lubke, The Peptides Vol. New York)] 1965 refers to the classic solution synthesis.

The above methods generally consist of adding one or more amino acids or appropriately protected amino acids to the growing peptide chain according to the sequence sequence. Typically, either the amino group or the carboxyl group of the first amino acid is protected with a suitable protecting group. The protected or derivatized amino acid is then coupled to either an inert solid support or used as a solution by addition of the following amino acid in the sequence, suitably protected at the additional (amino or carboxyl) group, under conditions suitable for the formation of the amide bond. The protecting group is then removed from the newly added amino acid residue and the next suitably protected amino acid is added and this operation is repeated. Finally, after all the desired amino acids have been linked in the appropriate order, any protecting groups (and solid support) may be sequentially or sequentially removed to give the desired polypeptide. By simple modification of the above general procedure, several amino acids can be added simultaneously to the growing chain, for example by coupling a protected tripeptide to a suitably protected dipeptide under conditions in which the chiral centers are not racemized and then deprotected to give a pentepeptide.

Preferably, the compounds of formula I are prepared by solid phase peptide synthesis.

In the particularly preferred process described above, the α-amino group of the amino acid is protected by an acid or base sensitive group. The requirement for the protecting groups is that they are stable under the conditions necessary for the formation of the peptide bond, but can be relatively easily removed from the growing peptide chain without damage or racemization of its chiral centers. Suitable protecting groups are, for example, tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), biphenylisopropyloxycarbonyl, tert-amyloxycarbonyl, isobornyloxycarbonyl1,1- dimethyl-3,5-dimethoxybenzyloxycarbonyl, 9-fluorenylmethyloxycarbonyl, or the like, especially tert-butoxycarbonyl being preferred.

Particularly preferred side chain protecting groups are as follows; arginine is nitro, p-toluenesulfonyl, 4-methoxybenzenesulfonyl, Cbz, Boc and adamantyloxycarbonyl; tyrosine is benzyl, isopropyl, cyclohexyl, cyclopentyl or acetyl; for serine, benzyl or tetrahydropyranyl; for histidine, benzyl, p-toluenesulfonyl or 2,4-dinitrophenyl.

The C-terminal amino acid is coupled to a suitable solid support. Suitable carriers for the above color thesis are materials which are inert to the reagents and reaction conditions during subsequent condensation and deprotection reactions and are insoluble in the medium used. Suitable solid carriers include chloromethyl polystyrene divinylbenzene polymer, hydroxymethyl polystyrene divinylbenzene polymer or the like, especially chloromethyl polystyrene-1% divinylbenzene polymer. In the particular case where the C-terminal end is glycinamide, a particularly preferred carrier is the benzhydrylaminopolystyrene divinylbenzene polymer [Rivaille P. et al., Hiv. Chim. Acta 54, 2772 (1972)]. For the chloromethyl-polystyrene-divinylbenzene-type resin, the N '-protected amino acid, preferably Boc-amino acid, is cesium, tetramethylammonium, triethylammonium, 1,5-diazabicyclo [5.4.0 ] undec-5-ene salt or the like in reaction with ethanol, acetonitrile, Ν, Ν-dimethylformamide or the like, it is particularly preferred to use the cesium salt in dimethylformamide at elevated temperatures, for example between 40 and 60 ° C, preferably At a temperature of about 50 ° C for 12 to 48 hours, preferably 24 hours. The N "-Boc-amino acid is coupled to the benzhydrylamine resin with Ν, Ν'-dicyclohexylcarbodiimide (DCC) and 1-hydroxybenzotriazole (HBT) for a reaction time of 2 to 24 hours, preferably 12 hours, at a temperature of from 50 to 50 ° C, preferably 25 ° C, in a solvent such as dichloromethane or DMF, preferably dichloromethane. The sequential coupling of protected amino acids can be carried out in an automatic peptide synthesizer well known to those skilled in the art. The N'protector7

Removal of the -7195234 powder is effected, for example, in the presence of trifluoroacetic acid solution in methylene chloride, hydrochloric acid in dioxane, hydrochloric acid in acetic acid or other strong acid solution, preferably 50% trifluoroacetic acid in dichloromethane at room temperature. . Each protected amino acid is preferably used in an excess of about 2.5 M and coupling is carried out in dichloromethane, dichloromethane-dimethylformamide, dimethylformamide or the like, preferably methylene chloride at room temperature. The coupling agent is usually DCC in dichloromethane, but it is also possible to use N, N'-diisopropylcarbodiimide (DIC) or other carbodiimide, alone or in the presence of HBT, N-hydroxysuccinimide, other N-hydroxyimide or oxime . Alternatively, protected amino acid active esters (e.g., ρ-nitrophenyl, pentafluorophenyl ester or the like) or symmetric anhydrides may be used.

Upon completion of solid phase synthesis, the protected polypeptide of each functional group is removed from the resin. When the polypeptide is linked to the resin by a benzyl ester-type bond, the cleavage of the polypeptide by aminoolysis with alkylamine or fluoroalkylamine on C-terminal peptides of proline, or with ammonia-methanol or ammonia-ethanol in peptides containing C-terminal glycine Aminolysis at 10 to 50 ° C, preferably 25 ° C, for 12-24 hours, preferably 18 hours. For example, the peptide may be cleaved from the resin by transesterification with methanol followed by aminolysis. At this stage of the procedure, the protected peptide can be purified by chromatography on silica gel. The side-chain protecting groups are removed from the peptide by treatment of the product obtained by aminolysis with, for example, anhydrous liquid hydrogen fluoride in the presence of anisole or other carbonium removal compound, or hydrogen fluoride-pyridine complex or boron tris (trifluoroacetate). and treated with trifluoroacetic acid or reduced with hydrogen in the presence of palladium on carbon or polyvinylpyrrolidone or treated with sodium in liquid ammonia, preferably liquid hydrogen fluoride in anisole at -10 ° C to 10 ° C, preferably the reaction time may range from 15 minutes to 1 hour, preferably about 30 minutes. Peptides containing glycine as the C-terminus are removed from the benzhydrylamine resin by cleavage and deprotection in a single step by treatment with liquid hydrogen fluoride in the presence of anisole as described above. The peptide obtained after deprotection is then purified by several successive chromatographic steps using the following 8 chromatographic steps: ion-exchange chromatography on a slightly basic acetate resin; hydrophobic adsorption chromatography on polystyrene-divinylbenzene (e.g. Amberlite XAD) resin-free; silica gel adsorption chromatography; ion-exchange chromatography on carboxymethylcellulose; partition chromatography, for example on Sephadex G-25 or countercurrent distribution; high performance liquid chromatography (HPLC), in particular reverse phase HPLC with octyl or octadecylsilyl silica gel bound phase column packing.

When a racemic amino acid is introduced at the 1-, 2-, 3- or 6-position, the final stereoisomeric nonapeptide or decapeptide product is separated and the desired D-amino acid peptide is isolated and purified, preferably by the chromatography method listed above. procedures.

Peptides containing aza-glycinamide at the C-terminus are preferably prepared by conventional solution phase peptide synthesis using known peptide intermediates. This procedure is described in detail in Example 3.

Accordingly, the present invention provides peptides of formula (I) and pharmaceutically acceptable acid addition salts thereof by deprotecting and optionally covalently bonding a solid polypeptide optionally bonded to a solid support corresponding to a compound of formula (I). carrier; or two fragments corresponding to the compound of formula (I) are joined in the appropriate order; and if desired,

a) converting a compound of formula I into a pharmaceutically acceptable acid addition salt; obsession

b) converting an acid addition salt of a compound of formula I into a pharmaceutically acceptable acid addition salt.

It should be noted that guanidino-substituted water-soluble amino acids, which may be substituted for the 6-position glycine in the process of the invention, are very important intermediates and are therefore an essential part of the invention.

Intermediates of general formula (II) wherein n is 4, R 1 is C 1 -C 6 alkyl, and R ₂ is C 1 -C 6 alkyl are preferred.

The above intermediates of formula (II) may be prepared by known methods, for example by guanylation or amidation of the corresponding amine.

The following examples are intended to illustrate the process of the invention without limiting it.

-8195234

Example A

Preparation of N-Acetyl-3- (2-naphthyl) -DL-alanine methyl ester

3- (2-Naphthyl) -DL-alanine is prepared as described in U.S. Patent 4,341,767.

The preparation of N-acetyl-3- (2-naphthyl) -DL-alanine and its conversion to the N-acetyl-3- (2-naphthyl) -DL-alanine methyl ester followed by separation of the D-isomer are also described above. patent.

Example B.

5.24 g of N-benzyloxycarbonyl-D-lysine benzyl ester-toluenesulfonate - as described by Bezus B. and Zervas L., [J. Chem. Soc., 83, 719 (1961)] and 1.72 ml of diisopropylethylamine in 60 ml of dioxane are treated with 1.89 g of Ν, N'-diisopropylcarbodiimide. The reaction mixture was stirred at 100 ° C for 6 hours, cooled to room temperature and evaporated to dryness. The solid residue is suspended in 20 ml of warm dimethylformamide, the Ν, Ν'-diisopropyl p.urea is filtered off and the filtrate is evaporated to dryness. Recrystallization from methanol / ethyl acetate gave N 'benzyloxycarbonyl-N, N'-guanidino-diisopropyl-D-homoarginine-benzyl ester-toluenesulfonate as an off-white solid.

[α] D = -7.26 ° (c = 0.3, methanol).

Similar to the above procedure, but in place of N, N-diisopropylcarbodiimide

Ν, Ν'-dicyclohexylcarbodiimide,

N, N'-di (n-hexyl) carbodiimide,

Ν, Ν'-diethylcarbodiimide,

N, N'-di (n-propyl) -carbodiimide,

N-isopropyl-carbodiimide,

N-propyl-carbodiimide

N, N'-di (n-butyl) carbodiimide,

N, N'-dicyclohexylcarbodiimide,

Ν, Ν'-diisobutyl carbodiimide,

N, N'-di (n-pentyl) -carbodiimide,

Ν, Ν'-diisopentyl carbodiimide

Ν, Ν'diphenyl carbodiimide

Ν, Ν'-ditolylcarbodiimide or

Using 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride or the like, the following compounds are prepared in the form of their sulfonate salts: N ^a -benzyloxycarbonyl-N, N'-guanidino -dicyclohexyl-D-homoarginine benzyl ester, [α] D = 8.07 ° (c = 0.9, methanol); N-benzyloxy-carbonyl-N, N'-guanidino-diethyl-D-homoarginine-benzyl ester; ^{N-benzyloxy-carbonyl-N,} N'-guanidino-di (n-propyl) -D-homoarginine-benzyl ester, [a] r> = - 8.07 ° (c = 0.9, MeOH ); ^Q -bcnzil N-benzyloxycarbonyl-N, N'-guanidino (n-propyl) -D-homoarginine-benzyl ester; ^{N-benzyloxy-carbonyl-N,} N'-guaniriino di (n-butyl) - D-homoarginine-benzyl ester; N ^a benzyloxycarbonyl-N, N'-guanidino-diisobutyl-D-homoarginine benzyl ester; N '-benzyloxycarbonyl-N, N'-guanidino-di (n-pentyl) -D-homoarginine benzyl ester;

N-benzyloxycarbonyl N, N'-guanidino-dimethyl-D-homoarginine benzyl ester,

^{N-benzyloxy-carbonyl-Ν,} N'-ethylguanidino-di (n-hexyl) -D-homoarginine-benzyl ester;

N '-benzyloxycarbonyl-5'-guanidine diisopropyl-D-homoarginine benzyl ester, [α] D = -10.5 ° (c 0.5, methanol); N-benzyloxycarbonyl-N-guanidino; (3-dimethylaminopropyl) -N'-guanidino-eth-D-homoarginine benzyl ester.

If instead of the D-lysine benzyl ester ^of N-benzyloxycarbonyl-D-ornithine benzyl ester are used to give the corresponding argininanalógokat form toluolszuifonátsó.

Example C II) ^B N-benzyloxy-carbonyl-N, N'-guanidino-D-homoarginine-ethano-benzyl ester mL of toluene and 15 ml of tert-butanol was added ^N-benzyloxy 2.71 g -carbonyl-D-lysine benzyl ester and 1.46 g of 2-methylthio-2-imidazoline hydrogen iodide (Aldrich) were added. The pH of the mixture was adjusted to 8 with diisopropylethylamine and the solution was refluxed for 24 hours.

The solution was concentrated in vacuo and the residue was applied to a silica gel column (250 g). The column was eluted by gradient elution with methylene chloride-methanol, varying the ratio of the ingredients from 19: 1 to 7: 3. The product-containing fractions were separated by thin layer chromatography, collected and evaporated to dryness to give 2.9 g of a white foam.

(ii) Preparation of N.N'-guanidino-ethano-D-homoarginine

2 g of this material are dissolved in 50 ml of ethanol containing 0.8 g of 10% palladium-on-carbon. The solution was stirred under a hydrogen atmosphere for 98 hours. The mixture was filtered through Celite and the filtrate was evaporated to dryness. 1.2 g of N, N'-guanidino-ethano-D-homoarginine are obtained in the form of a white foam.

(iii) Preparation of N-Boc-N, N'-guanidino-ethano-D-homoarginine (Compound 11a)

2.74 g of D-lysine di (hydrochloride) and 4.03 g of 2-methylthio-2-imidazoline-hydrogen iodide

A solution of 16.5 ml of 2N sodium hydroxide was stirred at room temperature for 6 days. Analysis of the reaction mixture in an amino acid analyzer showed that nearly 70% of the desired e-dialkylguanidino compound was formed. An additional 0.25 g of 2-methylthio-2-imidazoline hydrochloride iodide and 1 ml of 2N sodium hydroxide were added and the reaction was continued for 3 days at room temperature.

The reaction mixture was treated with 0.8 g of magnesium oxide ⁵ and 4.36 g of di-tert-butyl dicarbonate in 20 ml of dioxane. The pH is adjusted to 9.5 with 1N sodium hydroxide solution. After overnight reaction, there is still starting material in the reaction mixture, so another 1 g of di-tert-butyl dicarbonate is added.

-9195234

The mixture was filtered and the filtrate was evaporated to dryness. The residue was dissolved in water and washed with ethanol. The pH of the aqueous phase is adjusted to 4 with acetic acid. The acidic solution was washed with ethyl acetate. The aqueous phase containing the product was treated with anionic ion exchange resin (AG-3 acetate, BioRad) and evaporated to dryness.

The crude product was passed through a hydrophobic chromatography column (Amberlite XAD-2, Rohm and Haas) and eluted with a gradient of water to 25% ethanol. Fractions containing product were combined

2.7 g ^of N -Boc-N, N'-guanidino-D-homoarginine-ethano obtained, [a] D = -19.7 ° (c = 0.1, methanol).

By a similar procedure, instead of 2-methylthio-2-imidazoline hydrogen iodide

S-methyl diethylisothiocarbamide hydride iodide,

S-methyldiphenylisothiourea d-hydrogen iodide,

S-methyl-dibutyl-methylisothiourea hydriodide,

Using S-methyldipentylisothiourea hydroiodide or S-methyldihexyl 1-isothiourea hydroiodide the following compounds were prepared:

N "-Boc-N, N'-guanidino-diethyl-D-homoariginin,

N ^a -Boc-N, N'-guanidino-dipropyl-D-homoarginine,

N ^a -Boc-N, N'-guanidino-dibutyl-D-homoarginine,

N ^a Boc-N, N'-guanidino-dipentyl-D-homoarginine,

N "-Boc-N, N'-guanidino-dihexi 1-D-homoarginine,

Example D

Preparation of NMert-butoxycarbonyl derivatives of N, N'-guanidino-disubstituted-D-homoarginines from their toluenesulfonic acid salt precursors

A mixture of 3.25 g of N, N'-guanidino-diisopropyl-D-homoargininate toluenesulfonate and 100 mg of 10% palladium on carbon in 50 ml of glacial acetic acid was treated with hydrogen gas at atmospheric pressure for 4 hours.

Juk. The catalyst was filtered through a Celite filter and the acids were added resin, the following the filtrate evaporated to dryness according to solid N, N'-: 1, step CH2Cl2 wash ^- at 1 time 1.5 minutes Second step draft group removal 507 cf ₃ cooh / ch ₂ ci ₂ 1 time 1.5 minutes Third step draft group removal 507 CF 3 COOH / CH 2 Cl 2 1 time 30 minutes 4th step wash with CH ₂ Cl ₂ 3 times 1.5 minutes 5th step 107 triethylamine / CH 2 Cl 2 2 times 1.5 minutes 6th step wash with CH ₂ Cl ₂ 3 times 1.5 minutes 7th step N-Boc-amino acid solution 1 time dosage 8th step Ν, Ν'carbo-dicyclohexylcarbodiimide diimide 1 time dosage

-guanidino-diisopropyl-D-homoarginine toluenesulfonate is obtained. A solution of 2.13 g of the compound in 60 ml of 50% aqueous dioxane was added with 10 ml of 1 π sodium hydroxide and

Treat with 0.4 g of magnesium oxide. The mixture was then treated with 1.1 g of di-tert-butyl dicarbonate and stirred at room temperature for 2 hours. The magnesium salt was filtered off and the filtrate was concentrated in vacuo. The basic ¹ θ solution was washed with ethanol and then adjusted to pH 2.5 with sodium sulfate. The acidic solution was extracted with ethyl acetate and the extract was dried over anhydrous magnesium sulfate. The desiccant was filtered off and the filtrate was evaporated. Recrystallization from ethyl acetate-hexane gave N '- tert -butoxycarbonyl-N, N'-guanidino-diisopropyl-D-homoarginine toluenesulfonate.

In a similar manner, starting with the appropriate toluenesulfonate precursors can be prepared from ^the remaining N-tert-butoxycarbonyl-N, N'-guanidino-disubstituted-D-homoarginine or D-arginine.

²⁵ Example E

Boc-glycine (4.9 g) was dissolved in a mixture of ethanol (50 ml) and distilled water (50 ml). The solution was adjusted to pH 7 with aqueous cesium bicarbonate solution. The solvent was removed in vacuo.

The residue was dried under high vacuum for 18 hours and then dissolved in 150 ml of anhydrous dimethylformamide. 25 g of chloromethylated polystyrene - 1% divinylbenzene resin (Merrifield) (corresponding to 25 mmol of chloride) are added. The mixture was shaken at 50 ° C for 24 hours, filtered, and the resin washed successively with dimethylformamide, water and ethanol. The resin was dried under vacuum at ^{40 for} 3 days to give 28.34 g of Boc-Gly-O resin.

Example 1

7.29 g (5.5 mmol) of Boc-Gly-O resin prepared by reacting equimolar amounts of dry Boc-GlyOH cesium salt and chloromethyl polystyrene-1% divinylbenzene in a Beckman 990 peptide synthesizer reaction vessel. Synthesis of the prog50 with amino-10195234 was carried out sequentially

9th step CH ₂ Cl ₂ rinse, the 1 times switching rinse aid dispenser- reaction at 2 h outside, switching 10th step CH ₂ Cl ₂ rinse, dosage 1 time 1.5 minutes 11th step wash with CH ₂ Cl ₂ 3 times 1.5 minutes 12th step wash with ethanol 3 times 1.5 minutes 13th step wash with CH ₂ Cl ₂ 3 times 1.5 minutes 1-13. steps under one amino acid / 35¾ containing ammonium acetate

cycle, the reaction is performed by Kaiser E, et al., Anal. Biochem. 34, 595 (1970)].

The resin was coupled sequentially with 2.0-2.5 molar excess amino acids and DCC. Accordingly, the resin is subjected to successive coupling cycles

3.01 g with Boc-Pro-OH,

5.99 g of Boc-Arg (tosyl) -OH,

3.49 g of Boc-Leu-OH.H ₂ O are treated. The resulting tetrapeptide resin is then divided into smaller portions. At 1.0 g, the coupling cycles were continued with the following amino acids

0.456 g Boc-D-Dia-OH-toluenesulfonate

0.44 g Boc-Tyr (2,6-dichlorobenzyl) -OH,

0.375 g Boc-Ser (benzyl) -OH,

0.315 g Boc-D-Nal (2) -OH,

0.35 g Boc-D-p-F-Phe-OH,

0.275 g of Boc-L-proline and

2.5 ml of acetic anhydride.

The resin was removed from the reaction vessel, washed with dichloromethane and dried in vacuo. 1.66 g of a protected polypeptide resin are obtained. The protected peptide is separated from the resin by treatment with 50 ml of methanol saturated with ammonia at 0 ° C for 24 hours at room temperature. The resin was filtered off and washed successively with methanol and dimethylformamide. Removal of solvent from the filtrate in vacuo gave 0.9 g of the protected peptide as a yellow oil.

The protecting groups were removed by treatment with 10 ml of anhydrous liquid hydrogen fluoride in the presence of 1 ml of anisole in a Kel-F reaction vessel at 0 ° C for 30 minutes. The hydrogen fluoride was removed in vacuo and the resulting N-Ac-L-Pro-DpF-Phe-D -Nal (2) -Ser-Tyr-D-Dia-Leu-Arg-Pro-GlyNH ₂ -hydrofluoride was obtained. salt is washed with ether. The residue was then extracted with glacial acetic acid. The acetic acid extract was lyophilized to give 0.5 g of crude product.

The crude peptide was converted to the acetate salt by passing through a column packed with Ag3X (weakly basic tertiary amine resin) in acetate form in water. The eluate was lyophilized to give 0.5 g of the crude peptide as the acetate salt as a white solid.

The crude peptide was purified by HPLC using a Licroprep Rp-18 (25-40 microns) 2.5 x 100 cm column. The column was equilibrated at 0.03 mol /

methane and 65% water running buffer (pH 4.5). The main UV absorbing peak (280 nm), which elutes near the 3-fold column volume, is evaporated to dryness and lyophilized three times from distilled water. 75 mg of pure N-Ac-L-Pro-DPF-Phe-D-Nal (2) -Ser-Tyr-D-Dia-Leu-Arg-Pro-Gly-NH₂ peptide ₂ as a salt of acetic acid [j p ⁵ = -15 , 4 ° (c = 0.5 acetic acid).

In a similar manner, substituting the amino acids A, B, C or F with the above amino acids, the corresponding GlyNH ₂ decapeptides are prepared as follows: N-Ac-L-Pro-DpF-Phe-D-Nal (2) -Ser- Tyr-D-Dih-Leu-Arg-Pro-Gly-NH ₂ ,

N-Ac-L-Pro-DpF-Phe-D -Nal (2) -Ser-Tyr-D-Iph-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-D-Nal (2) -DpF-Phe-D-Nal (2) -Ser-Tyr-D-Iph-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Phe-Ser-Tyr-D-Iph-Leu-Arg-GlyNH ₂ ,

N-Ac-L-Pro-DpF-Phe-Trp-Ser-Tyr-D-Dih-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-L-Pro-DpF-Phe-D-Trp-Ser-Tyr-D-Dmh-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-L-Pro-DpF-Phe-DT rp-Ser-Tyr-D-Deh-Leu-Arg-Pro-GlyNh ₂ ,

N-Ac-L-Pro-DpF-Phe-D-Trp-Ser-Tyr-D-Dph-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-L-Pro-DpF-Phe-D-Trp-Ser-Tyr-D-Dhh-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-L-Pro-DpF-Phe-D-Trp-Ser-Tyr-D-Prh-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-DT rp-Ser-Tyr-D-Dmh-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-DT rp-Ser-Tyr-D-Deh-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dph-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-DT rp-Ser-Tyr-D-Dih-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dhh-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Prh-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Dmh-Leu-Arg-Pro-GlvNH ₂ ,

N-Ac-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Dph-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Nal (2) -DpF-Phe-D-Trp-Ser-Tvr-D-Dih-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Nal (2) -DpF-Phe-D-Trp-Ser-TdRD-Dhh-Leu-Arg-Pro-GlyNH ₂ ,

1

-11195234

N-Ac-Nal (2) -DpF-Phe-D-Nal (2) -Ser-Tyr-D-Dph-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Dhh-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Prh-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Hha-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dph-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dmh-Leu-Arg-Pro-GlyNH ₂ ,

N-Ac-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-GlyNH ₂ , and

N-Ac-Nal (2) -Dp-Cl-Phe-Trp-Ser-Tyr-D-dhh-Leu-Arg-Pro-GlyNH ₂ ,

Similarly, but using Boc-D-Ala-benzhydrylaminopolystyrene-1%-divinylbenzene resin instead of Boc-Gly-O-resin - which is made from benzhydrylaminopolystyrene-1% divinylbenzene resin (Beckman Inst), Boc-D- Prepared from Ala-OH, DIC and HBT, the following corresponding D-Alai ₀ -LHRH analogs were prepared:

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Nal (2) -Ser-Tyr-D-Iph-Leu-Arg-D-AlaNH ₂ ,

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dih-Leu-Arg-Pro-D-AlaNH ₂ ,

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dmh-Leu-Arg-Pro-D-AlaNH ₂ ,

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-D-AlaNH ₂ ,

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dhh-Leu-Arg-Pro-D-AlaNH ₂ ,

N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dia-Leu-Arg-Pro-D-AlaNH ₂ ,

N-Ac-D-Nal (2), -DP-Cl-Phe-D-Trp-Ser-Tyr-DDeh-Leu-Arg-Pro-D-AlaNH _2, [a] ₀ ²⁵ = -21.6 ° ( C = 0.4 acetic acid),

N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-D-AlaNH ₂ , N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dmh-Leu-Arg-Pro-D-AlaNH ₂ ,

N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dhi-Leu-Arg-Pro-D-AlaNH ₂ ,

N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dph-Leu-Arg-Pro-D-AlaNH ₂ ,

N-Ac-D-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-arg-Pro-D-AlaNH ₂ , m.p. 160-165 ° C, [a] p ° = -26.0 ° (c = 1.0, acetic acid)

Amino acid analysis:

Ser, 0.9 (1); Pro, 1.0 (1); D-Ala, 1.0 (1);

Leu, 1.0 (1);

Tyr, 1.0 (1); Trp, 0.9 (1); Arg, 1.0 (µ); p-F-Phe-, 0.9 (1);

D-Deh 0.9 (1); D-Nal (2), 0.9 (1); NH ₃ ,

FI ID.

N-Ac-D-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Dmh-Leu-Arg-Pro-D-AlaNH ₂ .

Example 2

Analogs containing the Pro-NHCH ₂ CH ₃ group at the C-terminus were prepared according to the synthesis program described in Example 1. 2.13 g of Boc-Pro-O-resin12, equimolar to solid Boc-Pro-OH-cesium salt and chloromethyl-polystyrene-1% divinylbenzene (Foot Systems, Inc.), were charged to the Beckman 990 synthesizer reaction vessel. by reacting quantities of the same. The above amount of Boc-Pro - O - resin

Contains 1.4 mmol of proline.

Resin in successive steps 2.0—

2.5 molar excess of protected amino acids and DCC. As stated above, the resin is reacted with the following amino acids in consecutive coupling cycles:

1.62 g Boc-Arg (tosyl) -OH,

0.93 g Boc-Leu-OH.H ₂ O,

0.94 g Boc-N, N'-guanidino-diisopropyl-hoarginine,

0.49 g of 1-hydroxybenzotriazole,

1.75 g of N-Boc-O-2-bromobenzyloxycarbonyl-L-triosine and

1.11 g Boc-Ser (benzyl) -OH.

At this point in the synthesis, the resin containing the protected polypeptide is divided into two parts and one part is reacted in successive steps with the following amino acids:

0.57 g Boc-D-Nal (2) -OH,

0.48 g Boc-D-p-F-Phe-OH,

0.21 g of N-Ac-L-proline.

The resin was removed from the reaction vessel, washed with dichloromethane and dried in vacuo. 2.26 g of the protected peptide resin are obtained.

The protected polypeptide was cleaved from the resin by aminolysis with 25 ml of ethylamine for 18 hours at 2 ° C. The ethylamine was allowed to evaporate and the resin was extracted with methanol. Evaporation of methanol gave 1.39 g of N-Ac-L-Pro-DpF-Phe-Trp-Ser (benzyl) -Tyr (2,6-dichlorobenzyl) -D-Dih-Leu-Arg (tosyl) -Pro- NHCH ₂ CH ₃ peptide was obtained. The protected peptide was stirred with 3 ml of anisole and 30 ml (from CoF ₃ ) of redistilled anhydrous liquid hydrogen fluoride at 0 ° C for 30 minutes in a Κθΐ-F reaction vessel. The hydrogen fluoride was removed in vacuo and the residue was washed with ether. The residue was dissolved in 2M acetic acid and lyophilized. 0.82 g of crude N-Ac-L-Pro-DpF-Phe-Trp-Ser-Tyr-D-Dih-Leu-Arg-Pro-NHCH ₂ CH _{3 is} obtained in the form of its acetic acid salt. The final purification was carried out by high performance liquid chromatography (20 mg)

2.5 x 100 mm, 40-50 micron octadecylsilylated purified by column chromatography with silica gel (Merck, Lichroprep C ₁₈₎ cartridge.

By proceeding in the same manner but with the appropriate protected amino acid at the appropriate site, the following compounds are prepared:

N-Ac-D-Nal (2) -D-p-F-Phe-D-Trp-Ser-D-Dih-Leu-Arg-Pro-NHEt,

N-Ac-D-Nal (2) -D-p-F-Phe-D-Trp-Ser-Tyr-D-Dph-Leu-Arg-Pro-NHEt,

N-Ac-D-Nal (2) -D-p-F-Phe-D-Trp-Ser-Tyr-D-Dmh-Leu-Arg-Pro-NHEt,

N-Ac-D-Nal (2) -D-p-F-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-NHEt,

-12195234

N-Ac-D-Nal (2) -D-p-F-Phe-D-Trp-Ser-Tyr-D-Dhh-Leu-Arg-Pro-NHEt,

N-Ac-D-Nal (2) -D-p-F-Phe-D-Trp-Ser-Tyr-D-Prh-Leu-Arg-Pro-NHEt,

N-Ac-D-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Hha-Leu-Arg-Pro-NHEt, and N-Ac-D-Nal (2) -DpF-Phe -D-Trp-Ser-Tyr-D-Dpa-Leu-Arg-Pro-NHEt.

In the above-described cleavage, ethylamine is substituted with stoichiometric amounts of methylamine and propylamine to produce the methylamides and propylamides of the nonapeptides listed above.

Example 3

Compounds of formula I wherein H is -NH-CONH are prepared by conventional solution synthesis.

An example of the preparation of the free peptide or salt of formula (5) is given in Scheme 1. AzaGlyNH ₂ in this case means -NH-NH-CONH ₂ .

The individual fragments are, for example, acyl azide (Honzel J. et al. Coll. Czecz. Chem. Comm. 26, 2333 (1971)],

It may be coupled by DCC / HBT coupling or other coupling process that does not cause racemization. The compound of formula (2) is known (Dutta A.S. et al., J. Chem. Soc. Perkin I, 1979, p. 379), and the compound of formula (1) can be prepared by the procedure described in Example 1. Compound (3) may be prepared from compound (2) by hydrogenolysis of the Cbz and nitro groups followed by D-Boc-N, N'-guanidino-diethyl-D-homorginin in the presence of DCC / HBT or other known condensing agents ( see Dutta et al., supra, for the preparation of similar LHRH analogs).

The product of Scheme 1 gives N-Ac-D-Nal (2) Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-Aza-GlyNH ₂ as the acetate salt. mp 153-155 ° C; [α] ²⁰ D = -29.6 ° (c = 0.3, acetic acid); amino acid analysis: Ser 0.9 (1); Pro 1.1 (1); Leu 1.0 (1); Tyr 0.98 (1); Trp 0.9 (1); D-Nal (2) 1.0 (1); Arg 1.0 (1); p-Cl-Phe 1.0 (1) D-Deh 1.0 (1); NH _? 1.3 (1).

Fragments linked in the above manner may be peptides or amino acids. The N-terminal nonapeptide acid may also be prepared by a solid phase or in solution process and then coupled to the semicarbazide hydrogen carbonate, dicyclohexylcarbodiimide hydroxybenzotriazole or other coupling procedure.

Example 4

A) 0.1 g of N-Ac-L-Pro-DpF-Phe-Trp-Ser-Tyr-D-Dia-Leu-Arg-Pro-GlyNH ₂ -hydrofluoride were prepared according to Example 1. Dissolve in water (50 ml) and pass through a solution of 50 g of Dowex 3 anion exchange resin, previously washed with acetic acid and washed with water. The column was eluted with deionized water and the eluate lyophilized. N-Ac-L-Pro-DPF-Phe-Trp-Ser-Tyr-D-Dia-Leu-Arg-Pro-Gly-NH ₂ acetic acid salt, [a] p ⁵ = 15.4 ° (C = 1, acetic acid).

By repeating the above procedure, but equilibrating the ion exchange resin with an acid other than acetic acid, such as hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, benzoic acid or the like, the corresponding salts are obtained.

Acid addition salts of other LHRH analogs prepared by the process of the invention can be prepared by a similar method.

For example, the hydrochloride salts are prepared as follows.

300 mg of N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-D-Ala-NH ₂ -hydrofluoride in 150 ml deionized dissolved in water and applied to a 1 x 10 cm column packed with Biograd Ag-3 (weakly basic) ion exchange resin previously equilibrated with 2N hydrochloric acid. The liquid passing through the column is recycled to the column and eluted with 150 ml deionized water. The eluate is lyophilized three times from deionized water. 210 mg of N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-D-Ala-NH ₂ -hydrochloride are obtained, [ a] p ⁵ = - 11.9 ° (c = 1, acetic acid), mp 166-170 ° C.

C) Preparation of the acid addition salt from free peptide N-Ac-L-Pro-DpF-Phe-D-Nal (2) -Ser-Tyr-D-Dia-Leu-Arg-Pro-Gly-NH _{2 in} free base form To this solution was added 30 ml of 1N acetic acid. The resulting solution was lyophilized to give 50 mg of the above peptide as an acetic acid salt. [α] ²³ = -15.4 ° (c = 0.5, acetic acid).

By proceeding in a similar manner, but employing acids other than acetic acid, the corresponding acid addition salts, such as hydrochloric, hydrobromic, sulfuric, phosphoric or nitric, are prepared in stoichiometrically equivalent amounts to the peptide.

Example 5 mg of a solution of N-Ac-L-Pro-DpF-Phe-D-Nal (2) -Ser-Tyr-D-Dih-Leu-Arg-Pro-Gly-NH ₂ -acetic acid salt in 25 mL of water 50 g of Dowex 1 (highly basic quaternary ammonium anion exchange resin) was loaded onto a column, which was previously equilibrated with sodium hydroxide to form hydroxyl ion. The column was eluted with 150 ml of water and the eluate was lyophilized. 45 mg of the corresponding polypeptide are obtained in the form of the free base.

In a similar manner, acid addition salts of other peptides of the invention, such as those described in Example 6, may be converted to the corresponding free base.

-13195234

Example 6

The following describes the preparation of some typical pharmaceutical compositions. The compositions contain as an active ingredient an LHRH analog of formula (I), such as N-Ac-L-Pro-DpF-Phe-D-Nal (2) -Ser-Tyr-D-Dih-Leu-Arg-Pro-Gly-NH ₂ , or a pharmaceutically acceptable acid addition salt thereof, such as an acetic acid salt.

- are included.

A) Preparation of tablets for oral administration (eg sublingual) LHRH Antagonist 10.0 mg compressible sugar, USP 86.0 mg Calcium stearate 4.0 mg LHRH Antagonist 10.0 mg compressible sugar 88.5 mg Magnesium stearate 1.5 mg LHRH Antagonist 5.0 mg Mannitol, USP 83.5 mg Magnesium stearate L5 mg Gelatinized starch 10.0 mg LHRH Antagonist 10.0 mg Lactose, USP 74.5 mg Gelatinized starch, USP 15.0 mg Magnesium stearate 1.5 mg Manufacturing process a) The LHRH antagonist in so much water

dissolve to mix with the sugar component to form a wet granulate mass. After thorough mixing, the granulate is dried in a tray or fluidized bed dryer. The dry granulate is screened to separate larger aggregates and then mixed with the other ingredients. The granulate is compressed into a tablet of suitable weight on a conventional tabletting machine.

b) This method is used to prepare lamellae containing 0.01% gelatin (USP). Gelatin is first dissolved in the granulation fluid and then the LHRH antagonist. Further steps are carried out as described in a).

Formulation 4 may also be used as an oral tablet.

B) Preparation of an Injectable Intramuscular Injection Formulation 1. An i.m. injectable preparation - sesame oil gel

LHRH antagonist 10.0 mg

Aluminum mono-stearate,

USP 20.0 mg

Sesame oil, q.s. To 1.0 ml

A mixture of aluminum monostearate and sesame oil was heated at 125 ° C with stirring to obtain a clear yellow solution.

The solution was then sterilized in an autoclave and allowed to cool. Under aseptic conditions, the LHRH analog is added while rubbing. It is particularly preferred to form LHRH analogs in the form of a slightly water-insoluble salt, such as zinc salts, zinc tannate salts, pamoate salts. The formulations containing the above salts have a particularly long duration of action.

2. Provided i.m. injectable preparation - biodegradable polymer microcapsule

LHRH Analog 1%

25/75 glycolide / lactide copolymer 99% (0.5 intrinsic viscosity)

Microcapsules (0-150) of the above composition are suspended in a vehicle of the following composition:

dextrose 5.0% sodium carboxymethylcellulose 0.5% benzyl alcohol 0.9%

Tween 80 0.1% Purified Water q.s. To 100.0% mg microcapsules are suspended in 1.0 ml of vehicle.

C) Aqueous solution for intramuscular injection

500 mg LHRH antagonist

Gelatin, no. antigen 5 mg

Water for injection q.s. To 100 ml

Gelatin and the LHRH antagonist are dissolved in water for injection and the solution is sterile filtered.

D) Preparation of Formulations for Rectal Administration

LHRH antagonist 5.0 mg

Witepsol H15 20.0 g

The LHRH antagonist is mixed with the molten Witepsol H15, homogenized and then poured into a 2 g mold.

Example 7 The biological activity of LHRH analog compounds was measured using the standard assay used to determine ovulation inhibition, Corbin, A. and Beattie, C.W. [Endocrin. Gap. Commun. 2 1 (1975) J.

-14195234

Compound Structure ED _SO (pg)

Propylene glycol corn brine oil

36 10 noon -24 hours noon -24 hours

UO

CN

OD salt CO f. 00 iO 00 r- CO CO CN CN

0 Γ. CO co 0 00 0 0 SHOOT F < Fi Λ Λ r. Λ Λ Fi Λ * CN τ- CN CN T CN She ·> - T SHE

CN

salt SHE Γ- » SHE r ** · Γ- SHE \SHE n n *> CN CN CN T · «- CN τ- CN 0

000 o

CD rh r-1 4-r f-1 1-1 co cj CO CD <3 1 1 1 1 4-r 4-r 1 1 1 r t 1 <3 <: <3 <3 P 1 1 1 1 P P P P X X RIP r> F \ F> RIP RIP Λ RIP * 43 43 43 42 43 43 43 • rl 4J • rí 42 42 p. ex Φ Φ P. Φ B 13 R P Φ Φ Q P P P P P P P P P P P P P P P P P P P P P P P E.g E.g E.g E.g E.g & fX E.g E.g cl cl cl μ E.g E.g E.g Pi Pi k Pi Pi Pi Pi Pi H E-i H H H H H H H E-i P P P P P P P P P P P P 1 She fiction Fi RIP e, »1 RIP Fi Φ RIP Fi Φ <u Φ G Φ Fi n Φ G 43 <U <u 43 43 43 43 43 0) Φ 43 43 E.g X 43 P Pi Pi Pi Pi 43 43 Pi Pi 1 P P 1 1 1 1 1 Pi E.g 1 | i-4 1 1 r-1 4-r 4-r 4-r r-1 1 1 t-1 r-l SHE PH pH u SHE SHE SHE u tx xH She SHE E.g ix E.g ix E.g E.g (X E.g E.g ix ex ex P P P P P P P Q Q Q P P Fi Fi 0) 0) ω <u G <u 43 Λ * 1 43 43 43 γΠ 43 F. fs Fi E.G, \ / -i PM E.g Pi Pi Pi / S z \ z s x s 1 CN CN 1 1 1 1 1 CN CN CN CN 4-r 'S 1-1 r-l 4-r ^ 4 RJ x_✓ S ^ z I υ 4-r r-l u 1 0 1 0 1 ζ? 1 0 1 r-l 1 1-1 1 1-1 1 r-l CJ 1 CJ CD CJ CD CJ 1 CJ 1 CJ 1 CJ 1 CJ 1 CJ CD G CD G CD G CD <3 IX <3 z <3 55 < ix <3 Pl <J ix <3 ex <3 ex <3 <3 <3 Z <3 X z P z P z P z P z P z P P 2 Q 1 Q 1 SHE 1 SHE z 1 She

-15195234

Example 8 Representative data of the compounds prepared according to the preceding Examples are summarized below.

Compound number

Structure of compound

1. [N-Ac-Dp-Cl-Phe ¹² , D-Trp ³ , D-Dih ⁶ , D-Ala ¹⁰ ] LHRH (second peak)

2. [N-Ac-Pro ¹ , DpF-Phe ² , D-Nal (2) ³ ,

D-monoisopropyl-Arg ⁶ ] LHRH

3. [N-Ac-Pro ¹ , DpF-Phe ² , D ₇ -Nal (2) ³ , D-Dia ^{e and} LHRH

4. [N-Ac-Pro ¹ , DpF-Phe ² , D-Nal (2) ³ ,

D-monoisopropyl-Arg ⁶ , Pro ⁹ -NHEt] LHRH

5. [N-Ac-Pro ¹ , DpF-Phe ² , D-Nal (2) ³ , D-Dia ⁶ ,

Pro ⁹ -NHEt] LHRH

6. [N-Ac-D-Nal (2) ¹ ' ³ ,. DpF-Phe ² , D-Dph ⁶ ] LHRH

7. [N-Ac-Pro ¹ , DpF-Phe ² , D-Nal (2) ³ , D-Dph ⁶ ] LHRH

8. [N-Ac-Dp-Cl-Phe ³ > ² , D-Trp ³ , D-Dph ⁶ , D-Ala ¹ °] LHRH

9. [N-Ac-Dp-Cl-Phe ^x > ² , D-Trp ³ , D-Dph ⁶ LHRH

10. [N-Ac-D-Nal (2) ¹ , D-pF-Phe ² , D-Trp ³ , D-Dph ⁶ ] LHRH (second peak)

11. [N-Ac-D-Nal (2) ¹ , DpF-Phe ² , D-Trp ³ , D-Dph ⁶ ] LHRH (third peak)

12. [N-Ac-Dp-Cl-Phe ¹ ' ² , D-Trp ³ , D-Dhh ⁶ D-Ala ¹ °] LHRH

13. [N-Ac-D-Ser ¹ -DpF-Phe ² , D-Trp ³ , D-Dhh ⁶ ] LHRH

14. [N-Ac-D-Nal (2) ^ DpF-Phe ² , D-Trp ³ , D-Dhh ⁶ ] LHRH

15. [N-Ac-Dp-Cl-Phe ¹¹² , D-Trp ³ , D-Deh ⁶ ] LHRH

16. [N-Ac-D-Nal (2) \ DpF-Phe ² , D-Trp ³ , D-Deh ⁶ ] LHRH

17. [N-Ac-Dp-Cl-Phe ¹ ' ² , D-Trp ³ , D-Det ^ D-Ala ¹ °] LHRH

18. [N-Ac-D-Nal (2) ¹ , Dp-Cl-Phe ² , D-Trp ³ , D-Deh ⁶ ] LHRH

19. [N-Ac-D-Nal (2) ¹ , Dp-Cl-Phe ² , D-Trp ³ , D-Deh ⁶ D-Ala ¹ °] LHRH

20. N-Ac-Dp-Cl-Phe ¹ ' ² , D-Trp ³ , D-Wsh ⁶ ] LHRH

21. [N-Ac-D-Nal (2) ¹ , DpF-Phe ² , D-Trp ³ , D-Wsh ⁶ ] LHRH

22. [N-Ac-Dp-Cl-Phe ¹ ' ² , D-Trp ³ , D-Dmh ⁶ , D-Ala ¹ ° 3LHRH

23. [N-Ac-D-Nal (2) ¹ , DpF-Phe ² , D-Trp ³ , D-Dmh ⁶ ] LHRH

24. [N-Ac-Dp-Cl-Phe ¹ ' ² , D-Trp ³ , D-DhPu-Ala ¹ °] LHRH

25. [N-Ac-D-Nal (2) ¹ -DpF-Phe ² , D-Trp ³ , D-Dhi ^s lLHRH

26. · [N-Ac-Dp-Cl-Phe ^{1 2} , D-Trp ³ , D-Dma ⁶ D, D-Ala ¹ °] LHRH

Physical constants:

1. Melting point: 157-160 ° C. ² Amino acid analysis: Ser, 0.82 (1); Pro, 1.12 (1); Ala, 1.14 (1); Leu, 55

1.16 (1); Tyr, 0.83 (1);

NH ₃ , 2.58; Arg, 0.88 (1); D-pCl-Phe + Trp,

2.34 (3); D-Dih, 1.15 0). '30

2. Melting point: i44-150 ° C fa] ²⁵ = -24.0 ° (c = 0.3 acetic acid)

Analysis for C ₇₃ H ₁₀₄ N | ₇ O | Calculated for C ₇ F (M: 1510.75): ⁶⁵ : C, 58.04; % H, 6.94; N, 15.76 = 15.76;

Found: C%, H% low

Amino acid analysis: Ser, 0.87 (1); Pro, 2.08 (2)

Giy, 1.01 (1); mono-iso- propyl Arg, 0.85 (I); Leu, 1.02 (1); Tyr, 0.90 (D; NH ₃ , 1.33 (1); Arg, 1.06 (1); D-pF-Phe, 0.91 (I); D-Nal (2), 1.09 (I). Third 144-148 ° C [< ⁵ -15.4 ° (e = 0.32 acetic acid) et emanalysis results for C ₇₆ H 1 10 ^ 17 o ₁₇ f together based on formula (M: 1552.83):

H, 7.14; N, 15.33. Found: C, H%: C, 58.78; H, 7.14; N, 15.33.

Amino acid analysis: Ser, 0.94 (1); Pro, 2.05 (2)

Gly, 1.00 (1); D-Dia, 0.85 (I); Leu, 1.03 (1); Tyr, 0.96 (1); NH ₃ , 1.27; Arg, 1.01 (1); D-pF-Phe, 0.95 (1); D-Nal (2), 1.06 (1);

4. Melting point: 145-150 ° C, ² [a] p ⁵ = -18.1 ° (c = 0.83, Acetic acid)

Amino acid analysis: Ser, 0.91 (1); Pro, 2.09 (2)

Leu, 1.05 (1); Tyr, 0.84 (1) NH ₃ , 0.41; Arg, 1.11 (1); D-pF-Phe, 0.91 (1); D-Nal (2); 0.95 (1); mono-isopropyl Arg, 0.74 (1).

5. Melting point: 137-142 ° C [a] £ ⁵ = -16.5 ° (c = 0.2, acetic acid) Elementary analysis for C ₇₆ H

111 N ₁₆ O

16 H (M: 1523.83): Calculated: C, 59.90; H, 7.34; N, 14.71;

Amino acid analysis: Ser, 0.91 (1); Pro 2.25 (2);

Leu, 1.05 (1); Tyr, 0.93 (1) NH ₃ , 0.67; Arg, 1.01; D-pF-Phe, 1.00 (1); D-Nal (2), 1.05 (1); D-Dia, 0.86 (1).

6. Melting point: 152-155 DEG C. [.alpha ⁵ - - Analysis I5,2 ° (c = 0.2, acetic acid) for C ₈₅ H _U6 N, O _{7) 7} based on molecular formula M (M: 1667):

Found: C, 61.24; H, 7.01; N, 14.28. Found: C, H% Low

Amino acid analysis: Ser, 0.94 (1); Pro, 1.11 (1)

Gly, 1.02 (1); Leu, 1.02 (1) Tyr, 0.90 (1); NH ₃ , 1.45 (1); Arg, 1.06 (1); D-pF-Phe, 0.94 (1); D-Dph,

1.06 (1); D-Nal (2), 2.32 (2).

7. Melting point: 135-142 ° C [j ²⁵ = -26.5 ° (c = 0.23, acetic acid) Elementary analysis for C ₇₇ H ₁₇ N _l2 O, F ₇ Calc'd (M: 1566.26) Found: C, 59.02; H, 7.20; N, 15.20 Found: C, H% Low

Amino acid analysis: Ser, 0.88 (1); Pro, 2.02 (2) Gly, 1.03 (1); Leu, 1.00 (1) Tyr, 0.88 (1); NH ₃ , 1.61; Arg, 1.08 (1); D-pF-Phe, 0.92 (1); D-Dph, 1.08 (1); D-Nal (2), 0.91 (1).

8. Melting point: 170-172 ° C [α] ²⁵ = -17.5 ° (c = 0.37, acetic acid)

Elemental Analysis Results a

C ₈₀ Hi ₁₄ Ni ₈ O, ₇ O ₁₇ Cl ₂ (M: 1670.82):

Calculated: C, 57.51; H, 6.88; N, 15.09 Found: C, H% Low

Amino acid analysis: Ser, 0.88 (1); Pro, 1.09 (1); Ala, 0.88 (1); Leu, 1.01 (1); Tyr, 0.92 (1); NH ₃ , 1.76; Trp, 0.41 (1);

1.19 (1); D-pCl-Phe, 1.84 (2); D-Dph, 0.97 (1).

Note: Trp is not completely separated from D-pCl-Phe.

9. Melting point: 165-170 DEG C. [.alpha ⁵ = -25 ° (c = 0.43, Acetic acid)

Elemental analysis (M: 1656.2) for C ₇₉ H ₁₂ N _E O ₇ Cl ₂ Calc'd Calculated: C% = 57.27, H% = 6.81, N = 15.22% Found: C% ', H% low

Amino acid analysis: Ser, (83 (1); Pro, 0.93 (1)

Ala., 1.01 (1); Leu, 1.02 (1) Tyr, 0.94 (1); NH ₃ , 1.18 (1); Trp, 0.97 (1) ^! ; Arg, 1.03 (1); D-pCl-Phe, 1.94 (2) ¹ ; D-Dph, 1.16 (1);

10. Melting point: 142-150 [deg.] C. [a] ⁵ opalescent solution in acetic acid

Elemental analysis for C ₈₂ N ₃ Hh _i8 O ₇ F chemical formula (m: 1641.93) Calculated: C% = 59.98, H% = 6.94, N% = 15.35% Found: C, H% low

Amino acid analysis: Ser, 0.95 (1); Pro, 0.93 (1)

Gly, 1.10 (1) ;; Leu, 1.09 (D Tyr, 0.87 (1); D-pF-Phe, 0.223 (1); NH ₃ , 2.87; Trp, 0.413 (1); Arg, 0.94 (1); D-Dph, 1.25 (1); D-Nal {2 (1.06 (1))

Note: D-pF-Phe values are low.

11. Melting point: 142-152 ° C ²

Amino acid analysis: Ser, 0.83 (1); Pro, 0.95 (1) Gly, 0.97 (1); Leu, 1.06 (1) Tyr, 0.96 (1); NH ₃ , 3.17: Trp, 0.66 (1); Arg, 1.01 (1 D-pF-Phe, 0.78 (1), D-Dph, 1.22 (1); D-Nal (2),

1.27 (1).

Melting point 140-142 ° C [α] ^D = -14.3 ° C (c = 0.14, acetic acid)

Elemental analysis results for C ₈₂ H | _l8 N _lg O | ₃ Cl ₂ . Based on 4 CH ₃ COOH.3H ₂ O (M: 1929.14):

Found: C, 56.03; H, 7.31; N, 13.07. C, 55.75; H, 6.93; N, 12.76

Amino acid analysis: Ser, 0.87 (1); Pro, 1.09 (1)

Ala, 1.08 (1); Leu, 1.02 (1) Tyr, 0.94 (1); NH ₃ , 1.43; Trp, 0.88 (l) ¹ ; Arg, 0.97 (1); D-pCl-Phe, 1.76 (2) '; Note: D-Dhh does not come off the column in 340 minutes.

13. Melting point: 145-148 ° C ~ 4.17 ° (c = 0.11, acetic acid)

Elemental Analysis for: C ₇₉ H ₁₁₂ N ₈ O ₁₈ F (M: 1629.96): Calculated: C 58.21, H 7.48, N 15.46 Found: C, H% low

Amino acid analysis: Ser, 1.77 (2); Pro, 1.15 (1)

Gly, 1.07 (1); Leu, 1.01 (1) Tyr, 0.96 (1); NH ₃ , 1.26;

-17195234

Trp, 0.73 (I); Arg, 1.01 D-pF-Phe, 0.81 (1).

14. mp: 147-151 DEG C. [.alpha ⁵ = -27.3 (c = 0.51, Acetic acid)

Elemental analysis for C ₈₉ H ₁₂ N | _{g of} O, F ₇ based on molecular formula (M: 1740.12) Calculated: C% = 61.43, H% = 7.36, N = 14.49% Found: C% H% low

Amino acid analysis: Ser, 1.00 (1); Pro, 1.11 (1)

Gly, 1.03 (1); Leu, 1.03 (1) Tyr, 0.95 (1); NH ₃ , 1.24 (1); Trp, 0.80 (1); Arg,

0.99 (1); D-pF-Phe, 0.87 (1); D-Nal (2), 0.85 (1).

15. Melting point: 155-158 ° C = -26 ° (c = 0.44, acetic acid)

Eiemanalysis results for C ₇₇ H | ₀₈ N ₁₈ O ₁₇ Cl ₂ based chemical formula (M: 1628.74) Calculated: C% = 56.78, H% = 6.68, N% = 15.48 found: C% H% low

Amino acid analysis: Ser, 0.88 (1); Pro, 0.97 (1)

Gly, 1.01 (1); Leu, 0.99 (1) Tyr, 0.97 (1); NH ₃ , 1.19; Trp, 0.926 (l) ¹ ; Arg, 1.03 (1); D-pCl-Phe, 1.85 (2) '; D-Deh, 1.28 (1).

Melting point: 152-155 ° C [α] ²⁵ D = -32.6 ° (c = 0.26, acetic acid)

Elemental analysis (Mn: 1628): Calculated for C ₈ N ₁₈ Ihi ,, 0, ₇ F Calcd Calculated:% C 59.76% H 6.87% N 15.49% Found: C, H % low

Amino acid analysis: Ser, 0.93 (1); Pro, 099 (1);

Glv, 1.10 (1); Leu, 094 (1) Tyr, 0.94 (1); NH ₃ , 1.19; Trp, 0.76 (1); Arg, 1.02 (1); D-pF-Phe, 0.92 (1); D-Deh, 1.23 (1); D-Nal (2) 0.95 (1).

Melting point: 155-157 ° C [α] ²⁵ = -18.4 ° (c = 0.27, acetic acid)

Elemental analysis results for C ₇₈ H | ₁₀ N ₁₈ O ₁₇ O ₁₇ Cl ₂ (M: 1642.76):

Calculated: C, 57.03; H, 6.75; N, 15.35. Found: C, H% Low

Amino acid analysis: Ser, 0.89 (1); Pro, 0.97 (1)

Ala, 0.98 (1); Leu, 1.03 (1) Tyr, 0.99 (1); NH ₃ , 1.34; Trp, 0.93 '; Arg, 1.00 (1); D-pCl-Phe, 1.86 (2) ¹ ; D-Deh, 1.17 (1).

18. mp 158-I60 ° C, [a] p ⁵ = -29.3 ° (c = 0.41, Acetic acid)

Elemental analysis for Cl ₈₁ H ₁ N ₁₈ O ₁₇ Cl (M: 1644.36):

Found: C, 59.16; H, 6.80; N, 15.33. Found: C, H% Low

Amino acid analysis: Ser, 0.92 (1); Pro, 1.02 (1)

Gly, 1.02 (1); Leu, 1.02 (I) Tyr, 0.99 (1); NH ₃ , 1.26; Trp, 0.83 (1); Arg, 0.98 (1); D-pCl-Phe, 0.98 (1); D-Deh, 1.16 (1); D-Nal (2) 0.96 (1).

19. Melting point: 154-157 ° C [α] ²⁵ = -21.6 ° C

Elemental analysis for C ₈₂ H ₁₃ N ₁₈ O ₁₇ CI (M: 1658.38):

Calculated: C, 58.39; H, 6.87; N, 15.20 Found: C, H% is low

Amino acid analysis: Ser, 0.95 (l); Pro, 1.08 (1)

Ala, 0.99 (1); Leu, 1.02 (1) Tyr, 1.01 (1); NH ₃ , 1.34; Trp, 0.85 (1); Arg, 0.97 (1); D-pCl-Phe, 1.18 (1); D-Deh, 1.17 (1); D-Nal (2) 1.00 (1).

20. Melting point: 150-155 ° C [α] D = -17.9 ° (c = 0.25, acetic acid)

Elemental Analysis for: C ₈₀ H ₁₁₅ N ₁₉ O ₁₇ Cl (M: 1685.84): Calculated: C, 56.99; H, 7.87; N, 15.79 Found: C, H % low

Amino acid analysis: Ser, 0.86 (1); Pro, 1.08 (1)

Gly, 1.04 (1); Leu, 0.97 (1) Tyr, 0.91 (1); NH ₃ , 1.32; Trp, 0.31 (1); Arg, 0.99 (1); D-pCl-Phe, 1.34 (2); Note: D-WSh does not come off the column in 340 minutes.

21. mp 1.50-154 DEG C. [.alpha ⁵ == -28.3 ° (c = 0.55, acetic acid) Elemental analysis for C ₈₄ H ₁₇ O _{I9 I18} N F Calcd (Mn: 1685):

Calculated: C, 59.88; H, 7.06; N, 15.79 Found: C, H% Low

Amino acid analysis: Ser, 0.87 (1); Pro, 0.94 (1)

Gly, 1.05 (1); Leu, 0.99 (1) Tyr, 0.95 (1); D-pF-Phe, 0.90 (1); NH ₃ , 1.34; Trp, 0.68 (1); Arg, 1.00 (1); D-Nal (2), 0.846 (1); Note: D-WSH does not come off the column in 340 minutes.

22. mp: 164-166 ° C [a] p ⁵ = -12.7 ° (c = 0.3, acetic acid) Elementary analysis for C ₇₆ Hi ₀₆ N ₁₈ O ₇ Cl ₂ Calc'd basis (Mn: 1,614, 72): Calculated: C, 56.53; H, 6.62; N, 15.61 Found: C, H% Low

Amino acid analysis: Ser, 0.93 (1); Pro, 1.07 (!)

Ala, 1.03 (1); Leu, 1.00 (1) Tyr, 0.97 (1); NH ₃ , 1.27; Trp, 0.95 (1) '; Arg, 1.11 (1); D-pCl-Phe, 1.9 (2) ¹ ; D-Dmh, 1.11 (1).

23. mp: 160-164 DEG C. [.alpha ⁵ = -26.5 ° (c = 0.29, Acetic acid)

Elemental analysis calculated for C ₇ gH ₁₀₇ F, ₈ F _l7 O Calc'd: C (Mn: 1,600): Calculated: C% = 59.31, H% = 6.74, N% = 15.76% Found: C, H % low

Amino acid analysis: Ser, 0.87 (1); Pro, 1.06 (1)

Gly, 1.00 (l); Leu, 1.02 (1) Tyr, 0.98 (1); NH ₃ , 1.31;

-18195234

Trp, 0.87 (1); Arg, 1.08 (l) ¹ , D-pF-Phe, 1.25 (I); D-Dmh 1.08 °; D-Nal (2), 0.89 (l).

24. mp: 160-165 ° C [a] D ~ ⁵ -12.2 ° (c = 0.29, acetic acid) Elementary analysis for C ₇₆ H | ₀₄ N | ₈ O _i7 basis Cl ₂ Calc'd (M: 1612.7) Calculated: C% = 56.60, H% = 6.50, N% = 15.60 found: C% H% low

Amino acid analysis: Ser, 0.94 (1); Pro, 1.14 (1) Ala., 1.01 (1); Leu, 0.97 (1); Tyr, 0.96 (1); NH ₃ , 1.23; Trp, 1.00 '; Arg, 1.06 (1); D-pCl-Phe, 2.00 (2) *; D-Dhi, 0.99 (I).

Melting point: 172-174 ° C [α] ²⁵ D = -29.8 ° (c = 0.62, acetic acid)

Elemental Analysis for: C ₇₉ H ₁₀ O ₅ N ₁₈ O ₇ F (M: 1598): Calculated: C, 59.38; H, 6.62; N, 15.78 Found: C, H; low

Amino acid analysis: Ser, 0.89 (1); Pro, 1.07 (1)

Gly, 1.00 (1); Leu, 0.99 (1) Tyr, 0.97 (1); NH ₃ , 1.24 (1); Trp, 0.85 (1); Arg,

1.04 (1); D-pF-Phe, 0.98 (2), D-Nal (2), 0.92 (1);

D-Dhi, 0.98 (l).

26. mp: 162-164 DEG C. _f] ²⁵ = -) - 18.7 ° (c = 0.54, acetic acid).

Elemental analysis for C ₇₅ H ₁₇ C 1 O _{I8 I04} N ₂ Calc'd (M: 1600.69) Calculated: C% = 56.28, H% = 6.55, N% = 15.75% Found: C, H% low

Amino acid analysis: Ser, 0.86 (1); Pro, 1.14 (1)

Ala., 1.01 (1); Leu, 1.01 (1) Tvr, 0.99 (1); NH ₃ , 1.54; Trp, 1.02 (l) ¹ ; Arg, 0.99 (1); D-pCl-Phe, 2.04 (2) ', D-Dma, 1.02 (1) ¹ .

Notes:

Ά peaks are not completely separated, the values given are the average of the two peaks.

² Optical rotation and / or elemental analysis was not performed due to lack of material.

The following LHRH analogs were also prepared: N-Ac-D-Nal (2) -Dp-Cl-Nal (2) -Ser-Tyr-D-Deh-Leu-Arg-Pro-D-AlaNH ₂ ; N-Ac-D-Nal (2) -DpF-Phe-D-Nal (2) -Ser-Tyr-D-Deh-Leu-Arg-Pro-D-AlaNH ₂ ;

Claims (23)

A process for the preparation of LHRH analogs of formula I wherein A is N- (C 2 -C 5) alkanoyl-Lprolyl; N- (C 2 -C 5) alkanoyl-D-p-chlorophenylalanyl, N- (C 2 -C 5) alkanoyl-D-seryl or N- (C 2 -C 5) alkanoyl-3- (2-naphthyl) -D-alanyl. B is D-p-chlorophenylalanyl or D-p-F-phenylalanyl group, C is D-tryptophanyl or 3- (2-naphthyl) -D-alanyl, F is an amino acid residue derived from a compound of formula II wherein n is 3 or 4; R 1 is C 1-6 alkyl; R ₂ is hydrogen, C 1-6 alkyl, or - (CH ₂ ) n - (R ₄ ) ₂ , where n is 1, 2, 3, 4 or 5 and R ₄ is 1-4. C 1 -C 4 alkyl; obsession R 1 and R _{2 taken} together form a ring of formula (f) H is D-alaninamide, glycinamide, or -NHR _{5 in} the latter formula R ⁵ is lower alkyl or -NHCONH ₂ - and pharmaceutically acceptable acid addition salts thereof, wherein j) deprotecting a protected polypeptide corresponding to a compound of formula I, optionally bonded to a solid support, and optionally removing the covalently bonded solid support; obsession b) linking the fragment corresponding to the compound of formula I in the appropriate order and optionally i) converting a compound of formula I into a pharmaceutically acceptable acid addition salt, or ii) converting an acid addition salt of a compound of formula I into a pharmaceutically acceptable acid addition salt. (Priority: June 9, 1983) 2. A process for the preparation of LHRH analogs of formula (I): wherein: A is N- (C2-C5) alkanoyl-L-prolyl, N- (C2-C5) alkanoyl-Dp-chlorophenyl, 1-, Ν-acetyl-D-seryl, or N- ( C 2 -C 5 alkanoyl-3- (2-naphthyl) -D-alanyl, B is D-p-chlorophenylalanyl or D-p-fluorophenylalanyl, C is D-tryptophanyl or 3- (2-naphthyl) -D-alanyl, F is an amino acid residue derived from a compound of formula II wherein n is 3 or 4; R 1 is C 1-6 alkyl; R ₂ is hydrogen, C 1-6 alkyl, or - (CH ₂ ) n - (R ₄ ) ₂ , where n is 1, 2, 3, 4 or 5 and R ₄ is 1-4. C 1 -C 4 alkyl; obsession R 1 and R _{2 taken} together form a ring of formula (f) -19195234 H is D-alaninamide, glycinamide, or -NHR _{5 in} the latter formula R ₅ is lower alkyl or -NHCONH ₂ - and pharmaceutically acceptable acid addition salts thereof, wherein a) deprotecting a protected polypeptide of formula I, optionally bonded to a solid support, and optionally covalently bonded solid support; obsession b) joining two fragments corresponding to the compound of formula I in the appropriate order and optionally i) converting a compound of formula I into a pharmaceutically acceptable acid addition salt, or b) converting an acid addition salt of a compound of formula I into a pharmaceutically acceptable acid addition salt. (Priority: 12/21/1982) 3. A process for the preparation of LHRH analogs of the formula I and their pharmaceutically acceptable acid addition salts comprising: A is N- (C 2 -C 5) alkanoyl-L-prolyl, N- (C 2 -C 5) alkanoyl-D-p-chlorophenylalanyl, N- (C 2 -C 5) alkanoyl-D-seryl, or N- (C 2 -C 5) is icanol 1-3- (2-naphthyl) -D-alanyl. B is D-p-chlorophenylalanyl, D-p-fluorophenylalanyl, C is D-tryptophanyl or 3- (2-naphthyl) -D-alanyl, F is an amino acid residue derived from a compound of formula II wherein n is 3 or 4, R 1 is C 1-6 alkyl? R ₂ is C 1 -C 6 alkyl or - (CH ₂ ) n -N (R ₄ ) ₂ , where R ₄ is C 1 -C 4 alkyl and n is 1, 2, 3, 4 or 5; and H is D-alaninamide, glycinamide, or -NHR _{5 in} the latter formula R ₅ is lower alkyl or -NHCONH ₂ - wherein a) deprotecting a protected polypeptide corresponding to a compound of formula (I), optionally in solid support, and optionally removing the covalently bonded solid support; obsession b) joining two fragments corresponding to the compound of formula (I) in the appropriate order; and, if desired i) converting a compound of formula I into a pharmaceutically acceptable acid addition salt; obsession b) converting an acid addition salt of a compound of formula I into a pharmaceutically acceptable acid addition salt. (Priority: June 10, 1982) A process according to claim 2 for the preparation of a compound of formula I or an acid addition salt thereof according to claim 2, wherein A is N-Ac-L-Pro, N-Ac-D-Ser, N-Ac-D-p-Cl-Phe or N-Ac-D-Nal (2), B is D-p-F-Phe or D-p-Cl-Phe, C is D-Trp or D-Nal (2), F is an amino acid residue derived from a compound of formula II wherein n is 3 or 4, R, methyl, ethyl, n-propyl, isopropyl or n-hexyl, and R ₂ is as defined in claim 1; obsession R 1 and R ₂ are 2-imidazolin-2-yl; H is D-AlaNH ₂ , GlyNH _2, or -NHEt, characterized in that the starting materials are the appropriate protecting group-optionally bonded peptides or the corresponding peptide fragments. (Priority: 12/21/1982) A process according to claim 4 for the preparation of a compound of formula I wherein A is N-Ac-L-Pro, N-Ac-D-Nal (2) or N-Ac-Dp-Cl-Phe, B is DpF-Phe or Dp-Cl-Phe, C is D-Nal (2) or D-Trp, F is D-Dmh, D-Deh, D-Dph, D-Dhh or D-Dhi and H is D -AlaNH ₂ , GlyNH ₂ or -NHEt, characterized in that the starting material is the appropriate protecting group, optionally bonded peptides or corresponding peptide fragments. (Priority: 12/21/1982) The process according to claim 5, wherein N-Ac-L-Pro-DpF-Phe-D-Nal (2) -Ser-Tyr-F-Leu-Arg-Pro-GlaNH is a narrower group of compounds of formula (I). ₂ , wherein F is D-Dmh, D-Deh, D-Dph, D-Dhh or D-Dhi; suitable peptide fragments are used. (Priority: 12/21/1982) 7. The process of claim 5, wherein N-Ac-L-Pro-D-ρ-Cl-Phe-D-Trp-Ser-Tyr-F-Leu-Arg-Pro- is a narrow group of compounds of formula I For the preparation of peptides of the formula NHE /, wherein F is D-Dmh, D-Deh, D-Dph, D-Dhh or D-Dhi, characterized in that the starting material is a corresponding protecting group, optionally bonded peptide or the corresponding peptide fragments are used. (Priority: 12/21/1982) -20195234 The process according to claim 5, wherein N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-F-Leu-Arg-Pro- is a narrow group of compounds of formula (I). GlyNH ₂ , wherein F is D-Dmh, D-Deh, D-Dph, D-Dhi, or Dhh, characterized in that the starting material is the appropriate protecting group, optionally supported peptides or peptide fragments are used. (Priority: 12/21/1982) The process a) or b) according to claim 3, which is N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dph-Leu. For the preparation of Arg-Pro-GlyNH ₂ , the starting material used is the appropriate protecting group optionally bonded peptide or the corresponding peptide fragments. (Priority: June 10, 1982) Process a) or b) according to claim 3, which is N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-GlyNH _2. characterized in that the appropriate starting materials are peptide moieties containing the protecting groups, optionally carrier-linked peptides or the corresponding peptide fragments. (Priority: June 10, 1982) The process according to claim 5, wherein N-Ac-D-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-F-Leu-Arg-Pro-GlyNH is a narrow group of compounds of formula (I). ₂ , wherein F is D-Dmh, D-Deh, D-Dph, D-Dhh or D-Dhi; suitable peptide fragments are used. (Priority: 12/21/1982) A process according to claim 3 for the preparation of N-Ac-D-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Dph-Leu-Arg-Pro-GlyNH _{2 according to} claim ₃ , characterized in that the starting material used is the appropriate protecting group, optionally bonded peptide or corresponding peptide fragments. (Priority: June 10, 1982) A process according to claim 3 for producing N-Ac-D-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Dph-Leu-Arg-Pro-GlyNH _{2 according to} claim ₃ , characterized in that the appropriate starting materials are peptide moieties or the corresponding peptide fragments which contain protecting groups. (Priority: June 10, 1982) 14. The process according to claim 5, wherein N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-F-Leu-Arg-Pro- is a narrow group of compounds of formula (I). Peptides of Formula D-AlaNH ₂ - wherein F is D-Dmh, D-Deh, D-Dph, D-Dhi, or D-Dhh, characterized in that the appropriate starting groups are peptide fragments containing appropriate protecting groups, optionally supported. (Priority: December 21, 1986) The process of a) or b) according to claim 3, N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Dph-Leu-Arg-Pro-D-AlaNH ₂ characterized in that the appropriate starting materials are peptide moieties or protecting peptides, which may be linked to a protecting group, or the corresponding peptide fragments. (Priority: June 10, 1982) The process a) or b) according to claim 3, wherein N-Ac-Dp-Cl-Phe-Dp-Cl-Phe-DT rp-Ser-Tyr-D-Deh-Leu-Arg-Pro-D-AliaNH ₂ characterized in that the appropriate starting groups are peptide moieties or appropriate peptide fragments which contain protecting groups. (Priority: June 10, 1982) 17. The process of claim 5, wherein N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-F-Leu-Arg-Pro is a narrow group of compounds of formula (I). -D-AlaNH ₂ peptides of the formula: - wherein D is D-DMH, D-Deh, D-Dph, D-DML or D-Dhi - preparation, characterized in that starting from the corresponding, optionally containing protective groups carrier bound peptides or appropriate peptide fragments. (Priority: 12/21/1982) A process according to claim 17 for the preparation of N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-D-AlaNH ₂ characterized in that the appropriate starting materials are peptide moieties or corresponding peptide fragments which contain protecting groups, optionally bound. (Priority: 12/21/1982) A process according to claim 3 a) or b) N-Ac-D-Nal (2) -Dp-Cl-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-GlyNH ₂ characterized in that the appropriate starting materials are peptide moieties containing the protecting groups, optionally carrier-linked peptides or the corresponding peptide fragments. (Priority: June 10, 1982) A process according to claim 5 for the preparation of N-Ac-D-Nal (2) -DpF-Phe-D-Trp-Ser-Tyr-D-Deh-Leu-Arg-Pro-D-AlaNH ₂ , using the appropriate protecting groups, optionally bonded peptides or corresponding peptide fragments, as starting materials. (Priority: 12/21/1982) 21. A process for the preparation of a pharmaceutical composition comprising the steps of: (1) The LHRH analogue of the formula -21195234, wherein A, B, C, F and H are as defined in claim 1, or a pharmaceutically acceptable acid addition salt thereof, is formulated with a pharmaceutical carrier and / or excipient. (Priority: June 9, 1983) 22. A process for the preparation of a pharmaceutical composition comprising the LHRH analogue of formula (I) as defined in claim 2, wherein A, B, C, E and H are as defined in claim 2, or a pharmaceutically acceptable salt thereof. the acid addition salt thereof in admixture with excipients and / or excipients conventionally used in the preparation of pharmaceuticals to form a pharmaceutical composition. (Priority: 12/21/1982) 23. A process for the preparation of a pharmaceutical composition comprising the LHRH analogue of formula (I) as defined in claim 3, wherein A, B, C, F and H are as defined in claim 3, or a pharmaceutically acceptable salt thereof. the acid addition salt thereof in admixture with excipients and / or excipients conventionally used in the preparation of pharmaceuticals to form a pharmaceutical composition.