HU211564A9 - Peptide compounds having therapeutic activity - Google Patents

Description

The invention relates to therapeutic (especially anti-nutrition) peptide compounds, to the use of the compounds as pharmaceutical and cosmetic therapeutic agents, and to compositions containing the compounds.

CCK-8, which has the structure: AspTyr(SO ₃ H)-Met-Gly-Trp-Met-Asp-Phe-NH _{2 ,} is known to have anti-feeding properties [see, for example, JE Morley, Minireview: The ascent of cholecystokinin from gut to brain', Life Sciences, 30 (6), 479—493 (1982)].

Published International Patent Application WO 91/08 225 (Fisons Corporation) describes several peptide compounds that are proposed for use in inhibiting feeding.

According to the present invention, there is provided a compound of formula (I) or a pharmaceutically acceptable derivative thereof (hereinafter collectively referred to as the "compounds of the invention"), wherein R ¹ is OH or OSO ₃ H:

M is Met, Ahx or Ile;

G is Gly or Sar;

X is Met, Ahx, Ile, Phe or Lys(R ² );

J is Asp. Asp(OBn), DAsp. MeAsp or MeDAsp;

L is Phe or MePhe;

R ² represents a group of general formula (II) in which

E is NH, CH=CH or CH ₂ CH ₂ group, and

R1, ^R4 , ^R5 , ^R6 and ^R7 are independently hydrogen, hydroxyl, halogen, C1-6 alkyl or _OSO3H ;

with the proviso that when R ¹ is a group of the formula OSO ₃ H, G is a Gly group and (a) J is Asp. L is MePhe and M is

Ahx. then X has a meaning different from Ahx;

(b) J is Asp. L is MePhe and M is

Ile. then X has a meaning other than Ile;

(c) J is Asp. L is MePhe and M is

Ile. then X has a meaning different from Ahx:

(d) J is Asp. L is Phe and M is Met. then X is other than Met;

(e) J is Asp. L is MePhe and M is

Met, then X is different from Met;

(f) J is DAsp. L is MePhe and M is

Met, X means different from Met.

The compounds of the invention, amino acids, peptides and protecting groups are designated by symbols commonly used in the art, for example those defined by IUPAC and IUB.

All optically active amino acids are L-configuration unless otherwise specified.

Examples of symbols include the following:

Ahx 2-aminohexanoic acid

Asn asparagine

Asp aspartic acid

Asp(Obn| aspartic acid-β-benzyl ester

Asp(OtBu) aspartic acid β-(tert-butyl) ester

5 Boc BrCH ₂ -Pam DAsp Fmoc Gly tert-butyloxycarbonyl 4-(4-bromomethyl)phenylacetamidomethyl D-aspartic acid 9-fluorenylmethyloxycarbonyl glycine 10 HOBt Hpa Hpa(SO ₃ H) Ile Lys 1 -hydroxybenzotriazolyl 4-hydroxyphenylacetic acid O-sulfo-4-oxyphenylacetyl isoleucine lysine 15 MeAsp MePhe MePhe-NH ₂ Met Nle N-methyl-aspartic acid N-methyl-phenylalanine N-methyl-phenylalanine amide methionine norleucine (or 2-aminohexanoic acid) 20 OBt 0CH ₂ Pam OSu * OtBu Phe 1 -benzotriazolyl ester 4-oxymethylphenylacetamidomethyl succinimidyloxy ester tert-butyl ester phenylalanine 25 Phe-NH ₂ resin Mud tBu Tyr(SO ₃ H) phenylalanine amide polystyrene sarcosine tert-butyl O-sulfotyrosine

Thus, Hpa(SO ₃ H)-Met-Gly-Trp-Met-Asp-MePheNH ₂ (A) is a compound of general formula (I) in which R ¹ is OH, M is Met, G is Gly, X is Met, J is Asp and L is MePhe.

The symbol Lys(R ² ) denotes a lysine group in which the ε-amino group forms an amide bond with a group of general formula (II) as defined above. Two specific groups of general formula (II) are mentioned, in one of which E is NH. R ³ is a methyl group and R ^{4 7} are each hydrogen, hereinafter referred to as "Tac"; and in the other group E is a trans-CH=CH group. R ⁵ is OH and R ³ . R ⁴ , R ⁶ and R ⁷ are all hydrogen.

Pharmaceutically acceptable derivatives of the compounds of formula (I) include esters and amides formed with any carboxylic acid group present and pharmaceutically acceptable salts. Among the pharmaceutically acceptable derivatives, unsubstituted amide derivatives of carboxylic acid groups (for example, Asp may be present as an unsubstituted amide derivative, i.e., Asn) and ester derivatives of carboxylic acid groups with C1-6 alkyl groups may be mentioned. Among the pharmaceutically acceptable salts, the sodium and ammonium salts are highlighted. Pharmaceutically acceptable derivatives of the compound of formula (1) can be prepared from the corresponding compounds of formula (I) by conventional methods.

The term "pharmaceutically acceptable" means that the compound, derivative, salt or other substance to which it refers is suitable for administration to the body as a medicinal or cosmetic therapeutic agent. Similarly, the terms "for use as a medicine and for

..medicinal product includes cosmetic7

HU 211 564 A9 also includes the use as a therapeutic agent, and the cosmetic therapeutic preparation.

Preferably, M is Ahx or Ile; G is Gly; X is Ahx, Ile or Lys(R ² ); J is Asp, DAsp, MeAsp or MeDAsp; and R ¹ is OSO ₃ H.

The invention further provides a process for the preparation of a compound of formula (I) or a pharmaceutically acceptable derivative thereof, which comprises a) sulfating a compound of formula (ΠΙ), wherein M, G and L are as defined above; Ja is as defined above for J, except that the β-carboxyl group of any of the groups Asp, DAsp, MeAsp or MeDAsp present is optionally protected; Xa is as defined above for X, except that it may also be Lys and any hydroxyl or amino group is present in protected form (except for the hydroxyl groups which are to be converted into a sulfate ester); and Za is NH ₂ or a carboxyl protecting group;

b) removing one or more protecting groups from a compound of formula (IV), wherein R ¹ , M, G, Xa, Ja and Za are as defined above, and at least one of Xa, Ja and Za is a protecting group;

c) reacting a compound of formula (V) wherein M, G, J and L are as defined above and Xb is Lys with a compound of formula (VI) wherein R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above to give a compound of formula (I) wherein X is Lys(R ² ) and E is NH; or

d) a compound of general formula (V), as defined above, is coupled with a compound of general formula (VII), in which E is CH ₂ CH ₂ or CH=CH and R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above, to give a compound of general formula (I) in which X is Lys(R ² ) and E is CH ₂ CH ₂ or CH=CH.

The invention also includes intermediate compounds of formula (IV).

In process a), the sulfating agent may be, for example, sulfur trioxide or a complex thereof, for example sulfur trioxide-pyridine. The sulfating is particularly preferably carried out in a polar aprotic solvent, for example dimethylformamide or pyridine. An excess of the sulfating agent, for example 1-40 molar, preferably 5 molar, is preferably used for the reaction.

The peptide protecting groups useful in processes a) and b) and the appropriate methods for their removal are well known in the art, for example from T. W. Greene, Protective Groups in Organic Synthesis (Wiley-Interscience, 1981). The choice of protecting groups and the methods used for their removal depend, inter alia, on the synthetic method used to prepare the peptide of formula (III) and the amino acids present in the peptide. Suitable amino protecting groups include the benzyloxycarbonyl group, which is readily removed by hydrogenolysis or acetic hydrobromide; the tert-butyloxycarbonyl group (Boc), which can be cleaved by allowing the peptide to stand in cold trifluoroacetic acid; Fmoc, which can be removed with dilute piperidine (20% in dimethylformamide); the (4-methoxybenzyl)oxycarbonyl and 2-nitrophenylsulphenyl groups. Boc and Fmoc groups are particularly preferred.

Suitable carboxyl protecting groups Za include, for example, methyl, tert-butyl, benzyl and 4-methoxybenzyl. The benzyl group is particularly preferred, as it can be easily removed by treatment with an alcoholic amine or ammonia. Similar groups can be used to protect the amino group in lysine and the carboxyl group in aspartic acid.

When the peptide is prepared by solid-phase methods, for example by coupling the carboxyl terminus of the peptide to a solid-phase resin, the peptide-resin linkage acts as a carboxyl protecting group. After cleavage of the peptide-resin bond, the carboxyl terminus of the compound of formula (II) is exposed. Since the sulfate ester-containing peptide end products of the invention are amides at the carboxyl terminus, the chemical bond between the peptide chain and the resin should be such that cleavage with an appropriate reagent yields the amide readily. The sulfate ester is labile in the presence of a strong acid (e.g., liquid hydrogen fluoride), and therefore the peptidyl-resin linkage should be cleavable either with weaker acids (e.g., by brief treatment with trifluoroacetic acid) and/or nucleophiles (e.g., ammonia, amines, hydroxides, and alkoxides).

Process c) can be carried out in an inert solvent, for example dimethylformamide, in the presence of a base, such as N-methylmorpholine, and at a temperature, for example, between 0 and 50°C.

Process d) can be carried out using an activated ester of the acid. A suitable active ester derivative is N-hydroxysuccinimidyl ester. The reaction can be carried out in the presence of a base, for example N-methylmorpholine, under the conditions described above for process c).

Suitable resin derivatives include, for example, oxymethyl polystyrene, 4-(oxymethylphenyl)-(CH ₂ ) _n -aminomethyl polystyrene (n - 0-3) and 4-(oxymethylphenyl)oxymethyl polystyrene. Similarly substituted polyacrylamide resins are as suitable as those based on polystyrene above. "Polystyrene" also includes copolymers containing small amounts, usually 1%, of unsaturated monomers, such as divinylbenzene.

4-(oxymethylphenyl) _-CH2CO -aminomethylpolystyrene [referred to herein as 4-(oxymethylphenyl)acetamidomethylpolystyrene or _OCH2- Pam resin] is particularly suitable for the preparation of peptide amides. This bond can be readily cleaved, as necessary, with methanolic ammonia, alkylamine or dialkylamine to form peptides of formula (I).

Another resin worth mentioning is a polystyrene resin (P), in which the linkage to the peptide is shown in formula (a) and in the description.

It is referred to as {{5-[(Fmoc-amino-methyl)-3,5-dimethoxy-phenoxy]-valeroyl}norleucyl)-4'-methyl-benzhydryl-amine-divinyl-benzene polystyrene or PAL resin. PAL resin is particularly advantageous for the preparation of peptide amides such as

HU 211 564 A9 resin, in which X is different from Lys. The bond between the assembled peptide and this resin can be easily cleaved with a mixture of trifluoroacetic acid, phenol, thioanisole, water and ethanedithiol in a ratio of 8.5:0.5:0.5:0.5:0.2.

Another resin that can be substituted for the PAL resin is a polystyrene resin (P) in which the linkage to the peptide is shown in formula (b), and the resin is hereinafter referred to as

4-[(2,4-Dimethoxyphenyl)-(Fmoc-amino)methyl]phenoxydivinylbenzene is referred to as polystyrene or Rink resin.

Peptides of formula (III), (IV) and (V) can be prepared by methods well known to those skilled in the art, for example by coupling individual amino acids stepwise on a solid-phase resin or by coupling groups of amino acids on a solid-phase resin to give the desired peptidyl resin intermediate. Such addition is carried out, as is known, by protecting the amino group of the amino acid or group of amino acids, for example by converting it into a tert-butyloxycarbonyl (Boc) or 9-fluorenylmethyloxycarbonyl (Fmoc) derivative, and then activating the carboxyl group of the amino acid or group of amino acids, for example by converting it into an ester derivative with hydroxybenzotriazole (HOBt) or N-hydroxysuccinimide (HOSu). Such protected-activated intermediates are then allowed to react with the free amino group of an amino acid resin or peptidyl resin, thereby extending the peptide chain and yielding the desired peptidyl resin.

The amino acid at the C-terminal position of the peptide to be produced can be coupled to the OCH2-Pam resin in several ways. For example, Boc-protected N-methylphenylalanine can be reacted with a suitable 4-(bromomethyl)phenylacetate ester (e.g. phenacyl ester), and the product can then be further converted to BocMePhe-(4-oxymethylphenyl)acetic acid, which can be coupled to aminomethylpolystyrene to yield Boc-MePhe-(4-oxymethylphenyl)acetamidomethylpolystyrene, Boc-Me-Phe- _OCH2 -Pam resin. We can also proceed by coupling 4-(bromomethyl)-phenylacetic acid to aminomethyl-polystyrene, thus forming 4-(bromomethyl)-phenylacetamidomethyl-polystyrene, BrCHi-Pam resin, which is reacted with the cesium salt of Boc-MePheOH to give Boc-Phe-OCH ₂ -Pam resin.

The C-terminal amino acid can be attached to the PAL resin by removing the Fmoc protecting group with a base, such as piperidine, in a suitable solvent or solvent mixture, such as dimethylformamide and toluene, followed by coupling of the protected activated amino acid in a manner conventional in solid phase synthesis. A particularly preferred method of carboxyl group activation is the preparation of the N-hydroxybenzotriazole (HOBt) ester in the presence of diisopropylcarbodiimide. A suitable solvent system, such as dimethylformamide and methylene dichloride, can be used for this preactivation process.

Suitable activating groups include any combination of groups that render the acid group of the amino acid more reactive, such as acid chlorides, mixed or symmetrical anhydrides, carbodiimide (e.g. dicyclohexylcarbodiimide,

with DCC), and the active esters, for example, esters derived from HOBt and HOSu, 2- or 4-nitrophenol and 2,4,5-trichlorophenol. The use of DCC and the esters of HOBt and HOSu is particularly advantageous in that no by-products are formed and consequently their purification is very simple.

For the solid phase synthesis of the sulfated peptides of the invention, an automated peptide synthesizer can be used. The peptides of formula (I) containing the sulfate ester can be desalted and purified by conventional methods. For example, the product can be chromatographed on Trisacryl M DEAE, DEAE-cellulose or similar ion exchangers, or partitioned on Sephadex LH-20, Sephadex G-25 or similar adsorbents,

Purification can be carried out by reverse phase chromatography on Amberlite XAD-2 (or Biorad SM-2), ODS-silica gel or similar adsorbent, normal phase chromatography on silica gel or similar adsorbent, or high-performance liquid chromatography (HPLC).

The protocol for coupling to aminomethyl resin or peptidyl-OCH ₂ Pam resin (1 mmol available nitrogen), deprotection, sulfation, cleavage and product purification is summarized in Table 1.

Table 1

Specification for the solid-phase synthesis of sulfated peptide amides on Parti resin (on the order of 1 mmol)

Step Reagent or solvent Purpose Mixing time 1 DCM Washing 1 minute 2 Continue with steps 3, 5 or 8 3 Weigh out a mixture of 3 mmol of protected amino acid or protected dipeptide, 4.5 mmol of HOBt and 3 mmol of DCC filtered and preactivated (0 °C. 1 hour) in 1:4 DMF/DCC Pre-engineered DCC/HOBi switching 2-15 hours

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Step Reagent or solvent Purpose Mixing time 4 Continue with steps 10,16,21 or 26 5 Add 3 mmol of protected amino acid or protected dipeptide and 4.5 mmol of HOBt in 30 ml of DMF/DCM 1:2, then 3 mmol of DCC in 20 ml of DCM In situ activated DCC/HOBt switching 2-15 hours 6 2-propanoic Washing 1 minute 7 Continue with steps 10, 16, 21 or 26 8 Weigh out 3 mmol of active ester or anhydride in DCM, DMF or a mixture of these solvents Non DCC/HOBt activated switching 2-15 hours 9 Continue with steps 10, 16, 21 or 26 10 DCM Washing 1 minute 11 Treatment with TFA/anisole/DCM 49:1:50 Boc and tBu removal 30 minutes 12 DCM Washing 1 minute 13 Treatment DIEA/DCM 1:19 mixture Neutralization 1 minute 14 DCM Washing 1 minute 15 Continue with steps 10, 16, 21 or 26 16 DMF Washing 1 minute 17 Treatment with a 1:4 mixture of piperidine/DMF Fmoc removal 3 minutes 18 Treatment with a 1:4 mixture of piperidine/DMF Fmoc removal 7 minutes 19 DMF Washing 1 minute 20 Continue with steps 10, 16, 21 or 26 21 DMF Washing 1 minute 22 Pyridine/DMF 1:2 Washing 1 minute 23 Weigh out 40 mmol of sulfur trioxide pyridine complex in 60 ml of pyridine/DMF 1:2 mixture Sulfation 20-24 hours 24 DMF Washing 1 minute 25 Continue with steps 10, 16, 21 or 26

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Step Reagent or solvent Purpose Mixing time 26 Methanol Washing 1 minute 27 250 ml of ammonia-saturated (-20 C) methanol or 20% methanolic amine solution Resin splitting 2-5 days 28 Methanol Washing 1 minute 29 Combine and evaporate the filtrates from steps 27-28 Separation 30 1 Chromatograph the residue on an Amberlite XAD-2 column (Rohm and Haas, 2.5 x 60 cm, methanol gradient in 0.1 M ammonia). Trisacryl M DEAE (LKB Inc., 2.5 X 47 cm, ammonium bicarbonate gradient) and/or P-40 ODS-3 (Whatman, 4.8 X 50 cm, methanol gradient in 0.2% ammonium acetate) Cleaning

Table 2

DIEA = diisopropylethylamine

A general description of the synthesis of non-sulfated peptide amides on PAL resin (1 g) is given in Table 2.

Step Reagent or solvent Purpose Mixing time 1 DCM Washing 1 minute 0 DCM/DMF (1:1) Washing 1 minute 3 30%- piperidine Fmoc 3 minutes 35% DMF removal 35% toluene 4 Repeat step 3 Fmoc removal 7 minutes 5 DCM/DMF (1:1) Wash (10X) 25 sec 6 Continue with step 7 or 11 7 Preactivated-protected amino acid [room temperature, Fniocaa-OH, 1 equiv DIPCD1, 1 equiv HOBt. in DMF:DCM( 1:4) Preactivation (If the previous amino acid MeAsp. MeDAsp or MePhe leaves the HOBt) 10 Minute Divorce ⁸ Add to resin or resin peptide Connection 1 -2 hours 9 DCM/DMF (1:1) Wash (4X) 1 minute 10 Continue with step 3 (Repeat steps 3-10 as needed to build the peptide sequence) 11 Preactivated Hpa-OSu [1 equivalent Hpa-OH, 1 equivalent HO-Su DMF] Pre-activation 12 Add to resin peptide Connection 1-2 hours ¹³ DCM Washing (4x) 1 minute 14 1_ MeOH Washing (4X) 1 minute

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Step Reagent or solvent Purpose Mixing time 15 TFA/phenol/thioanisole/water/ethanedithiol 8.5:0.5:0.5:0.5:0.2 Cleaving from resin 2-3 hours 16 TFA Rein filtration and washing 17 Combine and evaporate the filtrates obtained in step 16 Isolation of the peptide 18 Chromatate the residue on an Amberlite XAD-2 column (Biorad SM-2 200-400 mesh, methanol gradient in 0.1 M ammonia), and/or a C-18 ODS-3 column (Amicon, methanol gradient in 0.1% triethylamine, 0.1% glacial acetic acid) and/or an Amberlite XAD-2 column (methanol gradient) Cleaning 19 Combine the purified fractions and concentrate by freeze-drying from 0.1 M ammonia Separation

Practical modifications to the requirements of Tables 1 and 2 can be easily determined by experiment.

Similar processes in which the reaction is carried out without a solid phase component (resin) are also well known in the art and are well suited for large scale production (see, for example, U.S. Patent No. 3,892,726).

The compounds of the invention are capable of inhibiting feeding in mammals and of binding to cholecystokinin receptors. The different CCK receptors found in peripheral and brain tissues have been divided into the CCK-A and CCK-B receptors. Agonist and antagonist interactions with CCK receptors can be distinguished by functional studies. Activation of CCK-A receptors in peripheral tissues plays an important role in the regulation of pancreatic secretion, intestinal motility and gallbladder contraction. Compounds with agonistic activity against CCK-A receptors are used in the treatment of obesity and motility disorders. while compounds with antagonistic activity against CCK-A receptors are used in the treatment of gastrointestinal disorders. For example, they are used in the treatment of irritable bowel syndrome, ulcers, excessive pancreatic or gastric acid secretion, acute pancreatitis and motility disorders. Therefore, compounds that are intended to be used as therapeutic agents to inhibit feeding are likely to be free of undesirable side effects if they bind selectively to CCK-A receptors (rather than CCK-B receptors).

The therapeutic effect of the compounds of the invention can be demonstrated in the following tests A-D);

A) examination

Inhibition of feeding

Male Sprague-Dawley rats weighing 300-350 g were housed individually in cages. They were maintained on a 12-hour light: 12-hour dark cycle and were conditioned for at least 14 days to feed during a three-hour period of the dark cycle and not to feed for the 21 hours preceding the three-hour period.

On the day of the test, rats were given saline (control) or test compound dissolved in saline intraperitoneally, usually at doses ranging from 0.3 to 300 µg test compound/kg rat body weight. Food was given 10 minutes after saline or test compound administration. Test compounds were considered effective if the test group consumed significantly less food than the saline-treated control groups during the feeding period, which ended 0.5 or 3 hours after food introduction.

B) examination

CCK-A binding

The binding of the test compounds to CCK-A receptors in rat pancreatic membranes was measured against the binding of Bolton Hunter ¹²⁵ I-CCK-8 and -1H-L364,718 to rat pancreas according to the procedure described by Chang, Lotti, Chan and Kinkel (Molecular Pharmacology. 30: 212-216, 1986).

C) examination

CCK-B binding

The binding of the tested compounds to CCK-B receptors was measured in rat cerebral cortex membranes against ¹²⁵ I-CCK-8 using the method of Chang and Lotti (Soc. Natl. Acad. Sci. Vol. 83, 49234926).

D) examination

Functional study of CCK-A agonist/antagonist effects

To evaluate the ability of test compounds to inhibit or stimulate amylase release from rat pancreatic tissue fragments (acinar cells), the method of Lin et al. (J. of Pharm. and Exper. Therapeutics, 1986. 729-734) and Jung (Clinica Chemica Acta, 1980, 100. 7-11) was used.

Accordingly, the invention also relates to the use of the compounds of the invention as medicaments.

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The invention also extends to the use of the compounds of general formula (I) and their pharmaceutically acceptable salts for the preparation of pharmaceutical compositions suitable for the treatment of obesity.

The invention further relates to a method for treating obesity comprising administering a therapeutically effective amount of a compound of the invention to a patient in need thereof; and to a method for improving the physical appearance of a mammal comprising administering to the mammal a compound of the invention until a cosmetically desirable reduction in body weight occurs. The mammals of most interest are humans.

The invention also provides a pharmaceutical composition comprising a compound of formula (1) or a pharmaceutically acceptable salt thereof (preferably in an amount of less than 80% by weight, and more preferably less than 50% by weight) together with a pharmaceutically acceptable adjuvant, diluent or carrier.

The compounds of the invention can be administered by various routes, such as orally, intraperitoneally, intravenously, intramuscularly, subcutaneously, or intranasally. The dosage of the compounds of the invention will depend on several factors, including the requirements of the recipient, but will typically range from 0.3 pg to 3.0 mg/kg body weight per day, administered in a single dose or in divided doses.

Suitable adjuvants, diluents or carriers are, for example, lactose, starch, talc or stearic acid in the case of tablets or dragees; tartaric acid or lactose in the case of capsules; water, alcohols, glycerin or vegetable oils in the case of injection solutions.

The compositions may also contain suitable preservatives, stabilizing and wetting agents, solubilizing agents (e.g., a water-soluble cellulose polymer such as hydroxypropyl methylcellulose or a water-soluble glycol such as propylene glycol), sweetening, coloring and flavoring agents. The compositions may also be formulated in a sustained release form, if desired.

The compounds of the invention have the advantage of being more effective, more potent, longer acting, more stable, especially against enzymatic degradation, more selective, less toxic, cause fewer side effects, are better absorbed and act more rapidly or have other beneficial effects compared to the compounds of the prior art.

The invention is illustrated by the following examples, in which an automated peptide synthesizer was used for solid phase synthesis.

Example 1

Boc-MePhe-(4-oxymethylphenyl)acetic acid

To a solution of 27.93 g of Boc-MePhe-OH and 33.32 g of 4-(bromomethyl)phenylacetic acid phenacyl ester in 1000 ml of acetonitrile was added 18.28 g of potassium fluoride dihydrate. The suspension was stirred overnight, then filtered and the filtrate was evaporated to dryness. The residue, the

Boc-MePhe-(4-oxymethylphenyl)acetic acid phenacyl ester was dissolved in 1200 ml of 85% acetic acid, treated with 128 g of zinc powder and stirred for 2-4 hours. The reaction mixture was filtered, the filtrate was concentrated to a volume of about 400 ml and diluted with 3200 ml of water. An oil was then separated, which was dissolved in ethyl acetate and treated with dicyclohexylamine (DCHA) to give 41.31 g of the DCHA salt of the title compound, melting at 120-122 °C.

Example 2

H-MePhe-OCH ₂ -Pam resin

1.82 g (3 mmol) of BocMePhe-(4-oxymethylphenyl)acetic acid liberated from DCHA salt and 6.9 g (4.5 mmol) of HOBt in 40 ml of DMF:DCM = 1:3 by volume, then 0.62 g (3 mmol) of DCC in 20 ml of DCM solvent are added to 1.34 g (1 mmol of available nitrogen) of aminomethyl polystyrene resin and the resulting suspension is shaken for 2-15 hours. The Boc-MePheOCH ₂ -Pam resin is isolated by filtration, washed with 2-propanol and DCM, and then treated as indicated in Table 1 (steps 10-14) to give the title compound as the free base.

Example 3

H-Phe-OCH _r Pam resin

0.83 g (2 mmol) of Boc-Phe-(4-oxymethylphenyl)acetic acid, 0.46 g (3 mmol) of 1-hydroxybenzotriazole (HOBt) and 0.41 g (2 mmol) of DCC were dissolved in 50 ml of DCM:DMF - 4:1 and the solution was stirred at 0 °C for 1 h. 1.34 g of aminomethylpolystyrene resin (1 mmol of available nitrogen) was suspended in the reaction mixture, from which the precipitated dicyclohexylurea was removed by filtration and the mixture was shaken for 2-15 h. The product, Boc-Phe-OCH ₂ -Pam resin, was isolated by filtration and treated as indicated in Table 1 (steps 10-14) to give the title compound.

Example 4

Fmoc-Met-Asp(OtBu)-OH

FmocMet-OSu is prepared in situ by reacting 14.87 g of Fmoc-Met-OH, 5.52 g of HOSu and 8.26 g of dicyclohexylcarbodiimide in 200 ml of tetrahydrofuran at 0 °C for 3.5 hours.

The precipitated dicyclohexyl urea (DCU) is removed by filtration and the tetrahydrofuran filtrate is added to a cold solution of H-Asp(OtBu)-OH prepared in 220 ml of water:tetrahydrofuran - 10:1 volume ratio, to which 40 ml of 1 N sodium hydroxide solution has been added previously. The reaction mixture is stirred overnight at room temperature, then 20 g of solid citric acid and 600 ml of ethyl acetate are weighed out. The ethyl acetate layer is separated. with 10% citric acid solution. then washed with saturated aqueous sodium chloride solution and dried over magnesium sulfate. The ethyl acetal solution is evaporated, the residue is dissolved in 200 ml of ethyl acetate and reacted with 7.84 ml of DCHA. In this way, 17.93 g of material is precipitated. which is the DCHA salt of the title compound and melts at 159-162 °C.

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Example 5

The Hpa(SO _?l H)-Ahx-Gly-Trp-Ahx-Asp-Phe-NH ₂ H-Phe-OCH ₂ -Pam-resin, the product of Example 3, is coupled successively with Fmoc-Asp(OtBu)-OH, Fmoc-AhxOH, Fmox-Trp-OH, FmocGly-OH, Fmoc-Ahx-OH amino acids according to Table 1 (coupling steps 5-7, then steps 16-20 for removal of the Fmoc protecting group), thus obtaining H-Ahx-Gly-TrpAhx-Asp(OtBu)-Phe-OCH ₂ -Pam-resin, which is coupled with Hpa-OSu ester according to Table 1 (coupling steps 8-9), as the product, the resulting Hpa-Ahx-Gly-Trp-Ahx-Asp(OtBu)-Phe-OCH ₂ -Pam-resin after removal of the protecting group sulfated and cleaved from the resin according to Table 1 (steps 10-15, steps 21-25 and then steps 26-29 with ammonia) to give the title compound, which was purified by successive SM-2 and ODS-3 column chromatography according to Table 2 (step 18). Amino acid analysis after acid cleavage showed: Asp 1.03(1). Gly 1.05(1), Ahx 1.94(2), Phe 0.99(1), Trp 0.74(1), NH ₃ 2.16. MS(FAB): m/e 961 (MH) ⁺ .

Example 6

The protecting group is removed from the Hpa-Met-Gly-Trp-Met-DAsp-Phe-NH ₂ PAL resin, then the

According to Table 2, successively FmocPhe-OH. FmocDAsp(OtBu)-OH, Fmoc-Met-OH, Fmoc-Trp-OH, Fmoc-Gly-OH, FmocMet-OH amino acids and Hpa-OSu ester are coupled to give the title compound. The amino acid analysis after acid digestion showed: Asp 1.04(1), Gly 1.07(1), Met 1.79(2). Phe 1.10(1), Trp 0.71(1.0), NH, 1.40. MS (FAB): m/e 919 (M+H) ⁺ .

Example 7

The protective group is removed from the Hpa-Ahx-Gly-Trp-Ahx-Asp-Phe-NH ₂ PAL resin, then the

According to Table 2, successively coupled with FmocPhe-OH, FmocAsp(OtBu)-OH, Fmoc-Ahx-OH, FmocTrp-OH, Fmoc-Gly-OH, FmocAhx-OH amino acids and Hpa-OSu ester, thus obtaining the title compound. The amino acid analysis after acid digestion results: Asp 1.00(1), Gly 1.08(1.0), Ahx 1.85(2), Phe 1.10(1), Trp 0.68(1.0), NH ₃ 1.00. MS (FAB): m/e 883 (M+H) ⁺ .

Example 8

The protective group is removed from the Hpa-Ahx-Gly-Trp-Ahx-MeAsp-Phe-NH ₂ PAL resin, then the

According to Table 2, successively FmocPhe-OH, FmocMe Asp(OtBu)-OH, Fmoc-Ahx-OH, Fmoc-Trp-OH, Fmoc-Gly-OH, Fmoc-Ahx-OH amino acids and Hpa-OSu ester are coupled to give the title compound. The amino acid analysis after acid digestion showed: Gly 0.89 (1), MeAsp 1.04(1). Ahx 1.99(2), Phe 1.09(f), Trp 0.71(1.0), NH, 0.91. MS (FAB): m/e 897 (M+H) ⁺ .

Example 9

The protective group is removed from the Hpa-lle-Gly-Trp-lle-DAsp-Phe-NH ₂ PAL resin, then the

According to Table 2, successively FmocMePhe-OH, Fmoc-DAsp(OtBu)-OH, Fmoc-Ile-OH, Fmoc-Trp-OH, Fmoc-Gly-OH, Fmoc-Ile-OH amino acids and Hpa-OSu ester are coupled to give the title compound. The amino acid analysis after acid digestion showed: Asp 1.00(1), Gly 1.14(1), Ile 1.91(2), MePhe 0.96(1), Trp 0.64(1), NH ₃ 1.44. MS (FAB): m/e 897 (M+H) ⁺ .

Example 10

Hpa(SO _i H)-Met-Gly-Trp-Met-DAsp-Phe-NH ₂ 114 mg of the Hpa-Met-GlyTrp-Met-DAsp-Phe-NH ₂ peptide prepared according to Example 6 is dissolved in 1.4 ml of pyridine and 130 mg of sulfur trioxide-pyridine complex is added to the solution. The reaction mixture is stirred for 2 hours, then another 80 mg of sulfur trioxide-pyridine complex is added. After 4 hours, the reaction mixture is diluted with 20 ml of 5% ammonium hydroxide solution and then evaporated to dryness. The crude residue is purified by chromatography on SM2, ODS-3, SM-2 columns successively - according to step 18 of Table 2. The isolated product is freeze-dried from 0.1 M ammonia. Thus, 89 mg of the title compound is obtained. The amino acid analysis after acid digestion was as follows: Asp 1.03(1), Gly 1.11(1), Met 1.83(2), Phe 1.04(1), Trp 0.59(1), NH, 2.2. MS (FAB): m/e 997 (MH).

Example 11

Hpa(SOflj)-Ahx-Gly-Trp-A hx-MeA sp-Phe-NH ₂ 55 mg of the peptide Hpa-Ahx-GlyTrpAhx-Me Asp-Phe-NH ₂ prepared according to Example 8 was sulfated essentially as described in Example 10 to give 15 mg of purified title compound. The amino acid analysis after acid digestion was as follows: Me Asp 1.07(1), Gly 1.04(1), Ahx 1.90(2), Phe 0.98(1), Trp 0.61(1), NH ₃ 0.79. MS(FAB): m/e 975 (MH).

Example 12

Hpa(SO ₂ H)-Ile-Gly-Trp-Ile-DAsp-MePhe-NH ₂ 58 mg of the peptide Hpa-Ile-GlyTrp-Ile-DAsp-MePhe-NH ₂ prepared according to Example 9 was sulfated essentially as described in Example 10 to give 16 mg of purified title compound. Amino acid analysis after acid digestion showed the following: Asp 1.08(1), Gly 1.08(1), Ile 1.97(2), MePhe 0.87(1), Trp 0.75(1), NH ₃ 1.39. MS(FAB): m/e 975 (MH).

Example 13

Hpa-Met-Gly-Trp-Met-Asp-MePhe-NH ₂ The title compound was prepared by the sequential coupling of Fmoc-MePhe-OH, Fmoc-Asp(OtBu)OH, Fmoc-Met-OH. Fmoc-Trp-OH, Fmoc-Gly-OH, Fmoc-Met-OH and HpaOSu following the procedure described in Table 2. The amino acid analysis after acid digestion showed: Asp 1.06(1). Gly 1.04(1), MePhe 1.00(1),

HU 211 564 A9

Met 1.90(2), Trp 0.84(1), NH, 30.51. MS(FAB): m/e 931 (M-H).

Example 14

Hpa-Ahx-Gly-Trp-Ahx-DAsp-MePhe-NH ₂

PAL resin was coupled sequentially with Fmoc-MePhe-OH, Fmoc-DAsp(OtBu)-OH, Fmoc-Ahx-OH, Fmoc-Trp-OH, Fmoc-GlyOH, Fmoc-Ahx-OH amino acids and HpaOSu ester according to Table 2 to give the title compound. Amino acid analysis after acid digestion showed: Asp 1.03(1), Gly 1.04(1), Ahx 1.95(2), MePhe 0.99(1), Trp 0.77(1.0), NH ₃ 0.94. MS (FAB): m/e 895 (MH).

Example 15

Hpa-Ahx-Gly-Trp-lle-DAsp-Phe-NHs

PAL resin was coupled sequentially with Fmoc-Phe-OH, Fmoc-DAsp(OtBu)-OH, Fmoc-Ile-OH, Fmoc-Trp-OH, Fmoc-Gly-OH, FmocAhx-OH and HpaOSu ester according to Table 2. Thus, the title compound was obtained. The amino acid analysis after acid digestion was as follows: Asp 1.03(1), Gly 1.04(1), Ahx 0.96(1), Ile 0.98( 1). Phe 1.00( 1). Trp 0.67( 1), NH ₃ 0.99. MS(FAB): m/e 881 (MH).

Example 16

Hpa-Ahx-Gly-Trp-!le-MeAsp-Phe-NH ₂

PAL resin was coupled sequentially with amino acids Fmoc-Phe-OH, Fmoc-MeAsp(Ot Bu)-OH. Fmoc-Ile-OH, Fmoc-Trp-OH, Fmoc-Gly-OH, FmocAhx-OH and HpaOSu ester according to Table 2. This gave the title compound. The amino acid analysis following acid digestion showed: Me Asp 1.19(1). Gly 1.00(1), Ahx 1.02(1), Phe 0.91(1), Ile 0.89(1), Trp 0.65(1). NH, 0.60. MS(FAB): m/e 895 (M-H).

/ Example 7

Hpa-Ahx-Gly-Trp-lle-Asp-MePhe-NHs

PAL resin was coupled with amino acids Fmoc-MePhe-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Ile-OH, Fmoc-Trp-OH, Fmoc-Gly-OH, FmocAhx-OH and HpaOSu ester in succession according to Table 2. This gave the title compound. The amino acid analysis after acid digestion showed: Asp 1.10(1), Gly 1.09(1). Ahx 1.06(1), Ile 1.05(1), MePhe 0.71(1), Trp 0.76(1), NH ₃ 0.80. MS(FAB): m/e 895 (MH).

Example 18

Hpa(SO ₂ H/Ahx-Gly-Trp-Ile-DAsp-Phe-NH ₂ mg of the peptide Hpa-Ahx-Gly-Trp-Ile-DAsp-Phe-NH prepared in Example 15 according to Table 2 was sulfated essentially as described in Example 10 to give 22 mg of purified title compound. Amino acid analysis after acid digestion showed: Asp 1.07(1), Gly 1.04(1). Ile 0.97(1), Phe 0.80(1), Trp 0.46(1). NH, 1.88. MS(FAB): m/e 961 (MH).

Example 19

Hpa-Met-Gly-Trp-Met-As( OBn )-MePhe-NH ₂

Essentially following the procedures in Table 2 and sequentially coupling Fmoc-MePhe-OH, Fmoc-Asp(OBn)OH, Fmoc-Met-OH, Fmoc-Trp-OH, Fmoc-Gly-OH, Fmoc-Met-OH amino acids and HpaOSu ester, the title compound is prepared. MS(FAB): m/e 1023 (M+H) ⁺ .

Example 20

Hpa-(SO~,H i-Met-Gly-Trp-Met-Asp( OBn/MePheNH ₂

Peptide Hpa-Met-Gly-Trp-Met-Asp(OBn)-MePhe-NH ₂ , the product of Example 19, was sulfated essentially as described in Example 10 to give the purified title compound. Amino acid analysis after acid digestion showed: Asp 1.07(1), Gly 0.99(1), Met 1.91(2), MePhe 1.02(1), Trp 0.47(1). MS(FAB): m/e 1101 (MH)

Example 21

Hpa-Ahx-Gly-Trp-lle-DAsp-MePhe-NH ₂

Essentially following the procedure in Table 2 and coupling the amino acids Fmoc-MePhe-OH, Fmoc-DAsp(Ot Bu)-OH, FmocIle-OH. Fmoc-Trp-OH, Fmoc-Gly-OH, Fmoc-Ahx-OH and the Hpa-OSu ester, the title compound is prepared. The amino acid analysis after acid digestion showed: Asp 0.96(1), Gly 1.04(1), Ile 0.96(1), MePhe 1.07(1), Ahx 0.98(1), Trp 0.61(1), NH, 0.89. MS (FAB): m/e 895 (M-H).

Example 22

Hpa(SOyH)-Ahx-Gly-Trp-Ile-DAsp-MePhe-NH ₂

The peptide Hpa-Ahx-Gly-Trp-Ile-DAsp-MePhe-NHi. The product of Example 21 was sulfated essentially as described in Example 10 to give the title compound. Amino acid analysis after acid digestion showed: Asp 0.98(1), Gly 1.07(1), Ile 0.95(1), MePhe 1.41(1), Ahx 0.81(1). MS(FAB): m/e 975 (M-H), m/e 895 (M-SO,H).

Example 23

Hpa(SO ₂ H)-Ahx-Gly-Trp-lle-Asp-MePhe-NH ₂

The peptide Hpa-Ahx-Gly-Trp-Ile-Asp-MePhe-NH ₂ , the product of Example 17, was sulfated essentially as described in Example 10 to give the title compound. Amino acid analysis after acid digestion showed: Asp 0.98(1), Gly 1.08(1), Ile (0.96(1), MePhe 1.55(1). Ahx 0.84(1). MS(FAB): m/e 975 (MH), m/e 895 (M-SO,H).

Example 24

Hpa(SO ₂ H/Ahx-Gly-Trp-Ahx-DAsp-MePhe-NH ₂

The peptide Hpa-Ahx-Gly-Trp-Ahx-DAsp-MePhe-NH ₂ , the product of Example 14, was sulfated essentially as described in Example 10 to give the title compound. Amino acid analysis after acid digestion showed: Asp 1.00(1), Gly 0.98(1), MePhe 1.10(1), Ahx 1.93(2). Trp 0.96(1), NH, 0.30. MS(FAB): m/e 975 (MH).

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Example 25

Hpa-Ile-Gly-Trp-Ile-DAsp-Phe-NH ₂ Essentially following the procedure in Table 2 and coupling the amino acids Fmoc-Phe-OH, Fmoc-DAsp(OtBu)-OH, Fmoc-IleOH, Fmoc-TrpOH, Fmoc-Gly-OH, Fmoc-Ile-OH and the Hpa-OSu ester, the title compound is prepared. The amino acid analysis after acid digestion results in: Asp 1.02(1), Gly 1.03(1), Phe 1.01(1), Ile 1.94(2), Trp 0.72(1), NH ₃ 0.85. MS (FAB): m/e 881 (MH).

Example 26

Hpa(SO^H)-Ile-Gly-Trp-Ile-DAsp-Phe-NH ₂ The Hpa-Ile-Gly-Trp-Ile-DAsp-Phe-NH ₂ peptide, the

The product of Example 25 was sulfated essentially as described in Example 10 to give the title compound. Amino acid analysis after acid digestion showed: Asp 0.92(1), Gly 1.03(1), Phe 0.99(1), Ile 1.93(2). MS(FAB): m/e 961 (MH), m/e 881 (MSO,H).

Example 27

Hpa-Ahx-Gly-Trp-Phe-DAsp-MePhe-NHs Essentially following the procedure in Table 2 and coupling the amino acids Fmoc-MePhe-OH, Fmoc-DAsp(OtBu)-OH, FmocPhe-OH, Fmoc-Trp-OH, Fmoc-Gly-OH, Fmoc-AhxOH and the Hpa-OSu ester, the title compound is prepared. Amino acid analysis after acid digestion: Asp 1.01(1), Gly 0.99(1), Phe 1.06(1), Ahx 1.02(1), Trp 1.06(1), Phe 0.94(1), NH, 0.82. MS (FAB): m/e 931 (M-H).

Example 28

HpafSO^Hl-Ahx-Gly-Trp-Phe-DAs-MePhe-NHi The peptide Hpa-Ahx-Gly-Trp-Phe-DAsp-MePhe-NH ₂ , the product of Example 27, was sulfated essentially as described in Example 10 to give the title compound. Amino acid analysis after acid digestion showed: Asp 0.98(1), Gly 1.06(1), MePhe 0.97(1), Ahx 0.95(1), Trp 0.61(1), Phe 1.04(1), NH, 1.10. MS(FAB): m/e 1009 (MH). m/e 929 (MSO,H).

Example 29

Hpa(SOyH)-Ahx-Gly-Trp-Ahx-DAsp-Phe-NH ₂ Hpa-Ahx-Gly-Trp-Ahx-DAsp-Phe-NH ₂ peptide was prepared according to the method of Table 2 and sulfated essentially as described in Example 10. Amino acid analysis after acid digestion showed: Asp 1.02(1), Gly 1.05(1), Phe 1.02(1), Ahx 1.92(2), Trp0.81(1), NH, 1.48. MS(FAB): m/e 961 (MH), 881 (M-SO,H).

Example 30

Hpa(SCFH )-Ahx-Gly-Trp-Ile-MeAsp-MePhe-NHs Peptide Hpa-Ahx-Gly-Trp-Ile-MeAsp-MePhe-NH ₂ was prepared by the method of Table 2 and sulfated essentially as described in Example 10. The amino acid analysis following acid digestion showed:

MeAsp 0.92(1), Gly 1.00(1), MePhe 1.29(1), Ahx 0.77(1), Ile 1.02(1). MS(FAB): m/e 989 (MH), 909 (M-SO ₃ H).

Example 31

Hpa(SOyH)-Ile-Gly-Trp-Ile-MeAsp-Phe-NH ₂ Hpa-Ile-Gly-Trp-Ile-MeAsp-Phe-NH ₂ peptide was prepared according to the method of Table 2 and sulfated essentially as described in Example 10. Amino acid analysis after acid digestion showed: MeAsp 1.11(1), Gly 1.00(1), Phe 1.06(1), Ile 1.83(2), Trp 0.73(1), NH ₃ 1.48. MS(FAB): m/e 975 (MH), 897 (M-SO ₃ H).

Example 32

Hpa-Ahx-Gly-Trp-Ahx-MeDAsp-Phe-NH ₂ Essentially following the procedure in Table 2 and coupling the amino acids Fmoc-Phe-OH, Fmoc-MeDAsp(OtBu)-OH, FmocAhx-OH, Fmoc-Trp-OH, Fmoc-Gly-OH, Fmoc-AhxOH and the Hpa-OSu ester in sequence, the title compound is prepared. The amino acid analysis after acid digestion shows: MeAsp 0.78(1), Gly 1.10(1). Phe 1.05(1), Ahx 2.07(2), Trp 0.76(1), NH, 0.71. Ms(FAB): m/e 895 (MH) ⁺ .

Example 33

Hpa(SOfl-í )-Ahx-Gly-Trp-Ahx-MeDAsp-Phe-NH ₂ The peptide Hpa-Ahx-Gly-Trp-Ahx-MeDAsp-Phe-NH ₂ , the product of Example 32, was sulfated essentially as described in Example 10 to give the title compound. Amino acid analysis after acid digestion showed: MeAsp 1.00(1), Gly 1.02(1), Phe 1.06(1), Ahx 1.91(2), Trp 0.86(1), NH, 1.48. MS(FAB): m/e 975 (MH), m/e 896 (M-SO ₃ H).

Example 34

Hpa-Ahx-Gly-Trp-Ahx-MeAsp-MePhe-NH ₂ Essentially following the procedure in Table 2, the amino acids Fmoc-MePhe-OH, Fmoc-MeAsp(tOBu)-OH, Fmoc-Ahx-OH, Fmoc-Trp-OH, Fmoc-Gly-OH, FmocAhx-OH and the Hpa-OSu ester are coupled sequentially to the Rink-amide resin. The peptide is then cleaved from the resin using the usual procedure, and the 2-methylphenylacetamido group is formed on the E-amino group of Lys with o-tolyl isocyanate. The amino acid analysis following acid cleavage results in: MeAsp 1.01(1), Gly 1.05(1), MePhe 0.91(1), Ahx 2.02(2), Trp 0.84(1). NH, 0.82. MS(FAB): m/e 909 (MH).

Example 35

Hpa-Ahx-Gly-Trp-Lys(Tac)-Asp-MePhe-NH ₂ Essentially following the procedure in Table 2 and coupling the amino acids Fmoc-MePhe-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Ahx-OH, Fmoc-TrpOH, Fmoc-Gly-OH, FmocLys(Boc)-OH and the Hpa-OSu ester in sequence, the title compound is prepared. The amino acid analysis after acid digestion results in: Asp 1.01 (1), Gly 1.03( 1), MePhe 0.98( 1), Lys 0.94( 1). Ahx 1.04( 1). Trp 0.96( 1). MS(FAB): m/e 1046(MH) ⁺ .

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Example 36

Hpa(SOi,H)-Ahx-Gl\-Trp-Lys(Tac)-Asp-MePheNH ₂

The peptide Hpa-Ahx-Gly-Trp-Lys(Tac)-Asp-MePhe-NH ₂ , the product of Example 35, was sulfated essentially as described in Example 10 to give the title compound. Amino acid analysis after acid digestion showed: Asp 1.01(1), Gly 1.04(1), MePhe 0.92(1), Lys 0.93(1), Ahx 1.10(1), Trp 0.84(1). MS(FAB): m/e 1123 (MH) ⁺ .

Example 37

Hpa-Ahx-Gly-Trp-Ahx-Asp-MePhe-NH ₂

Following the procedure in Table 2 and coupling the FmocMePhe-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Ahx-OH, Fmoc-Trp-OH, Fmoc-Gly-OH, Fmoc-Ahx-OH peptides and the Hpa-OSu ester to the resin in sequence, the title compound was prepared. Amino acid analysis after acid digestion showed: Asp 1.00(1), Gly 1.02(1), MePhe 1.05(1), Ahx 2.1(2), Trp 0.53( 1), NH, 0.69. MS(FAB): m/e 897 (M+H) ⁺ .

Example 38

Hpa-Ahx-Sar-Trp-Ahx-Asp-MePhe-NH-.

Following the procedure in Table 2 and coupling the amino acids FmocMePhe-OH. Fmoc-Asp(OtBu)-OH, Fmoc-Ahx-OH, Fmoc-Trp-OH, Fmoc-Sar-OH, Fmoc-Ahx-OH and the Hpa-OSu ester sequentially to the resin, the title compound is prepared. The amino acid analysis after acid digestion showed: Asp 0.93( 1), Sar 1.12( 1), MePhe 0.95( 1), Ahx 1.99(2), Trp 1.06(1), NH, 0.88. MS(FAB): m/e 909 (MH) ⁺ .

Example 39

Hpa(SO ₂ H)-Ahx-Sar-Trp-Ahx-Asp-MePhe-NH-,

The peptide Hpa-Ahx-Sar-Trp-Ahx-Asp-MePhe-NH ₂ , the product of Example 38, was sulfated as described in Example 10 to give the title compound. Amino acid analysis after acid digestion showed: Asp 1.04(1), Sar 0.87(1), MePhe 0.86(1), Ahx 2.23(2), Trp 0.89(1), NH, 1.31. MS(FAB): m/e 989 (MH) ⁺ .

Example 40

The compound of Example 32 was subjected to the above test B) and found to have a binding constant (Ki) of 0.03 nM for binding to CCK-A receptors.

Claims (18)

PATENT CLAIMS 1. A compound of general formula (1), wherein R ¹ is OH or OSO,H; M is Met, Ahx or Ile; G is Gly or Sar; X is Met. Ahx. Ile. Phe or Lys(R ² ); J is Asp, Asp(OBn), DAsp, MeAsp or MeDAsp; L is Phe or MePhe; R ² represents a group of general formula (II) in which E is NH, CH=CH or CH ₂ CH ₂ group, and R1, ^R4 , ^R5 , ^R6 and ^R7 are independently hydrogen, hydroxyl, halogen, C1-6 alkyl or _OSO3H ; with the proviso that when R ¹ is a group of the formula OSO,H, G is a Gly group and (a) J is Asp, L is MePhe and M is Ahx, then X has a meaning different from Ahx; (b) J is Asp, L is MePhe and M is Ile, then X is different from Ile; (c) J is Asp, L is MePhe and M is Ile, then X has a meaning other than Ahx; (d) J is Asp, L is Phe and M is Met, then X is other than Met; (e) J is Asp, L is MePhe and M is Met, then X is different from Met; (f) J is DAsp, L is MePhe and M is Mel, X means different from Met; or a pharmaceutically acceptable derivative thereof. 2. A compound according to claim 1, wherein M is Ahx or Ile. 3. A compound according to claim 1 or 2, wherein X is Ahx, Ile or Lys(R ² ). A compound according to any one of the preceding claims, wherein J is Asp, DAsp, MeAsp or Me DAsp. 5. A compound according to any one of the preceding claims, wherein R ¹ is OSO,H. 6. According to claim 1 Hpa1SCLHi-Ahx-Gly-Trp-Ahx-Asp-Phe-NH,; Hpa-Met-Gly-Trp-Met-DAsp-Phe-NH ₂ ; Hpa-Ahx-Gly-Trp-Ahx-Asp-Phe-NH ₂ ; Hpa-Ahx-Gly-Trp-Ahx-MeAsp-Phe-NH ₂ ; Hpa-Ile-G]y-Trp-Ile-DAsp-MePhe-NH ₂ ; Hpa(SO,H)-Met-Gly-Trp-Met-DAsp-Phe-NH ₂ ; Hpa(SO,H)-Ahx-Gly-Trp-Ahx-MeAsp-Phe-NH ₂ ; Hpa(SO,H)-Ile-Gly-Trp-Ile-DAsp-MePhe-NH ₂ ; Hpa-Mel-Gly-Trp-Met-Asp-MePh-NH ₂ ; Hpa-Ahx-Gly-Trp-Ahx-DAsp-MePhe-NH ₂ ; Hpa-Ahx-Gly-Trp-Ile-DASp-Phe-NH ₂ ; Hpa-Ahx-Gly-Trp-Ile-MeAsp-Phe-NH ₂ ; Hpa-Ahx-Gly-Trp-Ile-Asp-MePhe-NH ₂ ; Hpa(SO,H)-Ahx-Gly-Trp-Ile-DAsp-Phe-NH ₂ ; Hpa-Met-GIy-Trp-Mel-Asp(OBn)-MePHe-NH ₂ ; Hpa(SO,H)-Met-Gly-Trp-Met-Asp(OBn)-MePHeNH ₂ ; Hpa-Ahx-Gly-Trp-Ile-DAsp-MePhe-NH ₂ ; Hpa(SO,H)-Ahx-Gly-Trp-Ile-DAsp-MePhe-NH ₂ ; Hpa(SO,H)-Ahx-Gly-Trp-Ile-Asp-MePhe-NH ₂ ; Hpa(SO,H)-Ahx-Gly-Trp-Ahx-DAsp-MePHe-NH ₂ Hpa-Ile-Gly-Trp-Ile-DAsp-Phe-NH ₂ ; Hpa(SO,H)-Ile-Gly-Trp-Ile-DAsp-Phe-NH ₂ ; Hpa-Ahx-Gly-Trp-Phe-DAsp-MePhe-NH ₂ ; Hpa(SO,H)-Ahx-Gly-Trp-Phe-DAsp-MePhe-NH ₂ ; Hpa(SO,H)-Ahx-Gly-Trp-Ahx-DAsp-MePhe-NH ₂ ; Hpa-Ahx-Gly-Trp-Ahx-DAsp-Phe-NH ₂ ; HU 211 564 A9 Hpa(SO ₃ H)-Ahx-Gly-Trp-Ile-MeAsp-MePhe-NH ₂ ; Hpa-Ahx-Gly-Trp-Ile-MeAsp-MePhe-NH ₂ ; Hpa(SO ₃ H)-Ile-Gly-Trp-Ile-MeAsp-Phe-NH ₂ ; Hpa-Ile-Gly-Trp-Ile-MeAsp-Phe-NH ₂ ; Hpa-Ahx-Gly-Trp-Ahx-MeDAsp-Phe-NH ₂ ; Hpa(SO ₃ H)-Ahx-Gly-Trp-Ahx-MeDAsp-Phe-NH ₂ ; Hpa-Ahx-Gly-Trp-Ahx-MeAsp-MePhe-NH ₂ ; Hpa-Ahx-Gly-Trp-Lys(Tac)-Asp-MePhe-NH ₂ ; Hpa(SO ₃ H)-Ahx-Gly-Trp-Lys(Tac)-Asp-MePheNH ₂ ; Hpa-Ahx-Gly-Trp-Ahx-Asp-MePhe-NH ₂ ; Hpa-Ahx-Sar-Trp-Ahx-Asp-MePhe-NH ₂ ; Hpa(SO ₃ H)-Ahx-Sar-Trp-Ahx-Asp-MePhe-NH ₂ ; or a pharmaceutically acceptable derivative thereof. A compound of formula (I) according to claim 1 or a pharmaceutically acceptable derivative thereof for use as a medicament. 8. A compound of formula (I) according to claim 1 or a pharmaceutically acceptable derivative thereof for use in the manufacture of a pharmaceutical composition for the treatment of obesity. 9. A pharmaceutical composition comprising a compound of formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof together with a pharmaceutically acceptable adjuvant, diluent or carrier. 10. A method for improving the physical appearance of a mammal, comprising administering to the mammal a compound of formula (I) according to claim 1 or a pharmaceutically acceptable derivative thereof until cosmetically beneficial body weight loss occurs. 11 A process for the preparation of a compound of general formula (I) or a pharmaceutically acceptable derivative thereof according to claim 1, which comprises a) sulfating a compound of general formula (III), in which M, G and L are as defined above; Ja is as defined above for J, except that the B-carboxyl group of any of the groups Asp, DAsp, MeAsp or MeDAsp present is optionally protected; Xa is as defined above for X, except that it may also represent Lys and any hydroxyl or amino group is present in protected form (except for the hydroxyl groups which are to be converted into a sulfate ester); and Za is NH _{2 or} a lauryloxy protecting group; b) removing one or more protecting groups from a compound of formula (IV), wherein R ¹ , M, G, Xa, Ja and Za are as defined above, and at least one of Xa, Ja and Za is a protecting group; c) reacting a compound of formula (V), wherein M, G, J and L are as defined above and Xb is Lys, with a compound of formula (VI), wherein R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above, to give a compound of formula (I), wherein X is Lys(R ² ) and E is NH; or d) a compound of general formula (V), as defined above, is coupled with a compound of general formula (VII), in which E is CH ₂ CH ₂ or CH=CH and R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above, thus obtaining a compound of general formula (I) in which X is Lys(R ² ) and E is CH ₂ CH ₂ or CH=CH. 12. An intermediate compound of general formula (IV) as defined in claim 1. 13. A substance or composition of matter comprising a compound of formula (I) according to claim 1 or a pharmaceutically acceptable derivative thereof for use in a method of improving the physical appearance of a mammal, the method comprising administering said substance or composition of matter to the mammal until cosmetically beneficial body weight loss occurs. 14. New compound, substantially as herein described. 15. A novel pharmaceutical composition, substantially as herein described. 16. A novel method for improving the physical appearance of a mammal, substantially as herein described. 17. A novel process for preparing a compound substantially as described herein. 18. A substance or composition for a new use in a treatment process, substantially as described herein.