AU660809B2

AU660809B2 - New peptide compounds having bradykinin-antagonist activity, process for their preparation and the pharmaceutical compositions which contain them

Info

Publication number: AU660809B2
Application number: AU41287/93A
Authority: AU
Inventors: Emmanuel Canet; Jean-Luc Fauchere; Nathalie Kucharczyk; Joseph Paladino; Christophe Thurieau
Original assignee: ADIR SARL
Current assignee: ADIR SARL
Priority date: 1992-06-18
Filing date: 1993-06-17
Publication date: 1995-07-06
Anticipated expiration: 2013-06-17
Also published as: CA2098655A1; EP0578521A1; DK0578521T3; DE69304155D1; JP2589937B2; FR2692581A1; EP0578521B1; DE69304155T2; ES2091570T3; ATE141615T1; FR2692581B1; GR3021472T3; JPH0687891A; NZ247914A; ZA934389B; AU4128793A

Abstract

Compounds of formula (I): <IMAGE> in which X, X', A1, A2, A3, A4, A5, A6, A7, A8, A 9 and Y are as defined in the description. Medications.

Description

IMUIU 11 2t//Ot Rogulaln 3.2(2)

AUSTRALIA

Patents Act 1990 660809

ORIGINAL

COMPLETE SPECIFICATION STANDARD PATENT Application Number: Lodged: o i Invention Title: NEW PEPTIDE COMPOUNDS HAVING BRADYKININ-ANTAGONIST ACTIVITY, PROCESS FOR THEIR PREPARATION AND THE PHARMACEUTICAL COMPOSITIONS WHICH CONTAIN THEM

S

The following statement is a full description of this invention, including the best method of performing it known to :-US I1 The present invention relates to new peptide compounds having bradykinin-antagonist activity, to a process for their preparation and to the pharmaceutical compositions which contain them.

Bradykinin, a natural nonapeptide, is known as a mediator of inflammatory and painful reactions, as well as in hypotensive states.

Bradykinin has, in addition, contractive effects on smooth muscle, especially that of the trachea, uterus or intestine.

In clinical medicine, bradykinin has been implicated in the physiopathology of shock states, inflammatory reactions, asthma and bronchial hyperreactivity, allergic or viral rhinitis, pancreatitis, arthritis, postgastrectomy dumping syndrome, psoriasis, hereditary angioneurotic-edema and migraine.

Bradykinin antagonists have' been described in the literature for the treatment of physiopathological situations in which high concentrations of bradykinin could be demonstrated, as in the case of septic shock (Robinson et al., Am. J. Med. 59-61, 2C 1975), arthritis (Shavura et al., Arch. Int. Pharmacodyn. 262- 279, 1983), asthma, allergic rhinitis and many other inflammatory and allergic reactions (Bradykinin antagonists: Basic and Clinical research (1991) ed. R. Burch, Marcel Dekker Inc., N. York).

This applies especially to the compounds described by Stewart S: and Vavrek in Patents WO 86/07,263 and WO 89/01,781, who demonstrated that the replacement of the proline at position 7 of bradykinin by a D-phenylalanine enabled an antagonist activity to be obtained. Patents EP 0,370,453 and EP 0,413,277 describe bradykinin antagonists obtained by replacement of the proline at position 7 by a D-phenylalanine or a tetrahydroisoquinoline-3-carbonyl residue and modification of the C- and Nterminal functions of the peptide.

The present invention describes bradykinin antagonists which are new not only in respect of the novel modifications of position A 7 of bradykinin and other amino acid substitutions in different positions, but also in respect of the introduction of substituents on the N- and C-terminal 2 functions of the peptide. These various modifications endow the compounds of the invention with especially intense pharmacological activities.

More specifically, the present invention relates to compounds of general formula

X\

X.A1-A2-A3-A4-A5-A6-A-A-A9-Y

(I)

in which: X .i substituent of the terminal amino group of the peptide of formula -whi.h -ay h i 1 -Ln. ir n, represents: a hydrogen atom, a 4-guanidinobenzoyl radical (Gba), unsubstituted or substituted on the phenyl ring with one or more halogen atoms or hydroxyl, mercapto, linear or branched (CI-C 6 alkyl, linear or branched (CI-C 6 alkylthio or linear or branched (CI-C 6 alkoxy radicals, a linear or branched (C 1 -Cs) alkyl radical, a (C 6

-C

1 2 aryl radical, a linear or branched (CI-C 6 alkoxycarbonyl radical, a linear or branched (Ci-C 6 alkylsulfonyl radical, a 9-fluorenylmethyloxycarbonyl radical, a linear or branched (CI-C 6 guanidinoacyl radical, unsubstituted or substituted with an amino radical, or a radical of formula /NH2 H2N-(CH2)n-CH \CO in which n is between 1 and 6,

A

1 and A9, which may be identical or different esent: a bond, ar te of formula 2a a substituent of the te rminal amino group of the peptidle of formula is hydrogen, A, represents: -a residue of formula see* 00* *00.0 0:* 3 R-(CH2)n-CH

\C

in which n represents an integer between 1 and 6 and RI an amino or guanidino radical, or any basic amino acid, unsubstituted or substituted on its side chain,

A

2 and A 3 which may be identical or different, represent: -a proline (Pro), hydroxyproline (Hyp), 2-azabicyclo[2.2.2]octane-3-carbonyl (Abo), 2-azabicyclo[2.2.l]heptane-3-carbonyl (Abh), homoproline (hPro), 'J,4-dehydroproline (dhPro), sarcosine (Sar), Nmethylphenylglycine (NMePgl), azetidine- 2-carbonyl (Azt) or alan.ine (Ala) residue,

A

4 represents Ae- a glycine (Gly) or sarcosine (Sar) residue or a residue of the formula: NH-(CH2)m-COin which m is an integer between 2 and 8,

A

5 represents a a pheriylalanine (Phe), fD-(2- *thienyl)alanine (Thi), D-c-c-lohexylalanine (Cha), P naphthylalanine (Nal), fD-furylalariine (Fur), fPpyranylalanine (Pra) or fP-pyridylalanine (Pyr) residue,

:A

6 represents any amino acid residue,

SA

7 any one of the following residues: N N N /N N N o/ CO/ Co /O co I I I II B C D E F -4

H

H- N- CH 3

CH

3 H- N N -CH 3

I

/N N N N CO N CO 0' Co Co N

/O

G H IJK L _N

N

C

3 N N 3 N COCO /N co /N N ,N I I M N o P Q R

RO

(R represents a linear or branched (Cl-C6) N, radical)

CO

S

or

:A-

7 represents a methylphenylalanine residue (MePhe) or a tetrahydroisoquinoline-3-carbonyl residue (Tic) (on condition that, in this case, X represents a pguanidinobenzoyl radical and XI a hydrogen atom),

A

8 represents a=~e-an octahydroindole-2-carbonyl (Oic) proline (Pro), hydroxyproline (Hyp), 2-azabicyclo(2.2.2]octane-3-carbonyl (Abo), 2-azabicyclot2.2.l]heptane-3-carbonyl (Abh), homoproline (hPro), 3,4-dehydroproline (dhPro), sarcosine (Sar), Nmethylphenylglycine (NMePgl), azetidine-2-carbony. (Azt) or alanine (Ala) residue or any one of the residues A to R defined for A 7 Y, a substituent of the terminal carbonyl grou ,f-ffepeptide of formula represents_.hfT- Y o "lor linear or branched (Cl-C 6 alk a ical or an amino radical D x(unsubs or substituted with one or two linear or Ag, a residue of formula

NH-

Ri-(CH2)n-CH

CO-

in which n represents an integer between 1 and 6 and R 1 an amino or guanidino radical, or any basic amino acid, unsubstituted or substituted on its side chain, Y, a substituent of the terminal carbonyl group of the peptide of formula represents a hydroxyl or linear or branched (Cl-C6) alkoxy radical or an amino radical (unsubstituted or substituted with one or two linear or e 9 0 *9 9* oo* 5 branched (Ci-C 6 alkyl radicals or phenyl radicals it being possible for each of the alkyl or phenyl radicals itself to be substituted with an amino, guanidino or ureido radical), or alternatively the carbonyl-terminal group of the peptide of formula is reduced to the corresponding alcohol (ol), and their addition salts with a pharmaceutically acceptable acid or base, on the understanding that: when X represents a p-guanidinobenzoyl radical, X' a hydrogen atom, AI an arginine residue (Arg), A 2 a proline residue (Pro), A 3 a hydroxyproline residue (Hyp), A 4 a glycine residue (Gly), A 5 a P-(2-thienyl)alanine residue (Thi), A 6 a serine residue (Ser), As an octahydroindole-2carbonyl residue (Oic), Ag an arginine residue (Arg) and Y a hydroxyl radical, then A 7 is other than a tetrahydroisoquinoline-3-carbonyl residue (Tic) of D or L configuration with respect to its a carbon, each peptide bond -CO-NH- of the peptide of the formula can be optionally replaced by a pseudopeptide bond chosen from -CH 2 -NH, -CH 2

-CH

2

-CH

2

-SO

2

-NH-CO-

or -CH=CH-, each amino acid of the peptide sequence being optically pure and the a carbon of each amino acid having the D or L configuration.

S Among pharmaceutically acceptable acids, there may be mentioned, hydrochloric, hydrobromic, sulfuric, phosphonic, acetic, trifluoroacetic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, tartaricr maleic, citric, ascorbic, methanesulfonc and camphoric acids, and the like.

6 Among pharmaceutically acceptable bases, there may be mentioned, sodium hydroxide, potassium hydroxide, triethylamine, tert- butylamine, and the like.

The expression i"any amino acid" used in the present application includes both natural amino acids and so-called non-proteinogenous amino acids which are commonly used by a person skilled in the art in the synthesis of synthetic analogs of natural peptides.

Among natural amino acids, there may be mentioned, glycine, alanine, valine, leucine, isoleucine, serine, methionine, threonine, phenylalanine, tyrosine, tryptophan, cysteine, proline, histidine, aspartic acid, asparagine, glutamine, glutamic acid, arginine, ornithine and lysine.

Among non-proteinogenous amino acids, there may be mentioned, norleucine, norvaline, alloisoleucine, homoarginine, thiaproline, dehydroproline, hydroxyproline, homoserine, cyclohexylglycine, phenylglycine, amino-n- butyric acid, cyclohexylalanine, aminophenylbutyric acid, pheriylalanines Smono-, di- or trisubstituted on the phenyl group at the ortho, meta or para position with group(s) chosen from (Ci-C 6 alkyl,

(CI-C

6 alkoxy, halogen, trifljoromethyl, nitro, amino, monoor di(C 1

-C

6 alkyl)amino or alternatively a methylenedioxy group, and 3- thienyl)alanine, and 3furyl)alanine, 3- and 4-pyridyl)alanine, and 3benzothienyl)alanine, and 2-naphthyl)alanine, O-alkyl derivatives of serine, threonine, and tyrosine, Salkylcysteines, all amino acids of D configuration, isomers of natural amino acids of L configuration, and the like.

The invention also extends to the process for preparing the compounds cf formula which may be obtained by various methods such as sequential solid- phase synthesis, the synthesis of fragments and their coupling in solution, S enzymatic synthesis and genetic synthesis by cloning and expression of genes in transformed bacteria, or by various combinations of these techniques.

The general methods of solid-phase peptide synthesis have been described by B.W. ERICKSON and R.B. MERRIFIELD ("The Proteins", Solid-Phase Peptide Synthesis, 3rd edition, 257- 527, 1976).

The solid-phase synthesis may be carried out on an automated apparatus which performs in a repetitive and programmable manner deprotection, coupling and washing cycles needed for the sequential introduction of the amino acids into the peptide chain. The amino acid, preferably C-terminal, is attached to a resin conventionally used for the preparation of polypeptides, preferably a polystyrene crosslinked using to 3.0 of divinylbenzene and equipped with activated residues such as chloromethylene or hydroxymethylene which enable the first amino acid to be attached covalently to the resin. The appropriate choice of resin enables a C-terminal carboxylic acid, amide or alcohol function to be formed after synthesis.

The amino acids are then introduced one by one in the order Sdetermined by the operator. Each synthesis cycle corresponding to the introduction of an amino acid entails a deprotection, preferably N-terminal, of the peptide chain, successive washes designed to remove the reactants or to swell the resin, a coupling with activation of the amino acid and further washes.

Each of these operations is followed by a filtration, accomplished by means of the presence of a glass sinter incorporated in the reactor in which the synthesis takes place.

The coupling reagents used are standard reagents of peptide synthesis, such as dicyclohexylcarbodiimide (DCC) and hydroxybenzotriazole (HOBT) or benzotriazol- 1yloxytris(dimethylamino)phosphonium hexafluorophosphate (BOP) S or alternatively diphenylphosphoryl azide (DPPA).

Activation by mixed anhydride formation is also possible.

Each amino acid is introduced into the reactor in approximately four-fold excess with respect to the degree of substitution of the resin, and in an approximately equivalent amount with respect to the coupling agents. The coupling reaction may be checked at each step of the synthesis by the 8 ninhydrin reaction test described by E. KAISER et al. (Analyt.

Biochem., 34, 595, 1970).

After assembly of the pept3 chain on the resin, treatment with a strong acid such as trifluoroacetic acid or hydrofluoric acid in the presence of anisole, ethanedithiol or 2-methylindole serves to separate the peptide from the resin and also to free the peptide, where appropriate, from its protective groups. The compound is then purified by standard purification techniques, in particular chromatographic techniques.

The peptides of the present invention may also be obtained by the coupling in solution of selectively protected peptide fragments, which may be prepared either on a solid phase or in solution. The use of protective groups and the means of taking advantage of their differential stability are similar to the solid-phase methods, except for the attachment of the peptide chain to the resin. The C-terminal carboxyl g.oup is protected, for example, by a methyl ester or an amide function. The methods of activation during coupling are also similar to those employed in solid--phase synthesis.

The synthesis of peptides containing pseudopeptide bonds such as -CH2-NH-, -CH 2

-CH

2

-CH

2

-SO

2 -NH-CO- or -CH=CHis performed either by solution methods or in a combined procedure with solid-phase synthesis using standard methods of organic chemistry. Thus, for example, the introduction of the S -CH 2 -NH- bond is accomplished by preparing in solution the aldehyde Fmoc-NH-CHR-CHO according to the technique described by FEHRENTZ and CASTRO (Synthesis, 676-678, 1983) and condensing it with the growing peptide chain, either on a 306". solid phase according to the technique described by SASAKI and COY (Peptides, 8, 119-121, 1988), or in solution.

The compounds of formula possess very advantageous pharmacological properties, especially bradykinin-antagonist properties. On this basis, they may be used beneficially in a number of therapeutic indications such as traumas, grazes, burns, skin eruptions, eczema, erythema, edema, sore throat, arthritis, asthma, allergies, rhinitis, anaphylactic shock, inflammations, arterial hypotension, pains, pruritus and insufficiency of spermatozoal mobility.

The subject of the present invention is also pharmaceutical compositions containing as active principle at least one compound of general formula or one of its addition salts with a pharmaceutically acceptable acid or base, alone or in combination with one or more non-toxic, inert excipients or vehicles.

Among the pharmaceutical compositions according to the invention, there Imay be mentioned, more especially, those which are suitable for oral, parenteral or nasal administration, simple or sugar-coated tablets, sublingual tablets, sachets, packets, hard gelatin capsules, suppositories, creams, ointments, skin gels, aerosols, ampoules containing preparations to be swallowed or injected, etc.

The dosage varies according to the patient's age and weight, the nature and severity of the disorder and also the administration route.

The latter can be oral (including inhalation and sublingual administration), nasal, rectal, parenteral or transdermal.

Generally speaking, the dosage ranges between 0.1 pg/kg and 5 mg/kg for a treatment administered in one or several doses per 24 hours.

The examples which follow illustrate the invention and in no way limit it.

In the examples below, the amino acids whose abbreviations begin with a capital letter are of the L configuration.

The amino acids whose abbreviations begin with a lower-case letter are of the D configuration.

The abbreviations used in the examples are as follows: S Gba in place of 4-guanidinobenzoyl, Arg in place of the arginine residue, Pro in place of the proline residue, dhPro in place of 3,4-dehydroproline, Hyp in place of the hydroxyproline residue, Gly in place of the glycine residue, Thi in place of the -(2-thienyl)alanine residue 10 Ser tic amc 2-c] ttp Dpr Gpr Dbu Sar Aoc ttp ttp [2,3-cl tfp tfp 2-c] tfp 3-c] tpp 3, 4-c] tpp [3 tpp (2m] tpp .9 ~c? 4.

4 9.

44 4 94 4.

44.44 9 4 *9 .4 4 4 499* ~'4 99 4 4 90 94 '444 **4 4 3t~ in place of the serine residue, in place of the tetrahydroisoquinoline-3-carbony1 residue, in place of the octahydroindole-2-carbonyl residue, in place of the (6R)-4,5,6,7-tetrahydrothieno[3,2c]pyridine-6-carboxylic acid residue.

in place of the 2,3-diaminopropionic acid residue, in place of the 2-amino-3-guanidinopropionic acid residue, in place of the 2,4-diaminobutyric acid residue, in place of the sarcosine residue, in place of the 8-aminooctanoic acid residue, in place of the (6R)-4,5,6,7-tetrahydrothieno[3,4clpyridine-6-carboxylic acid residue, in place of the (6R)-4,5,6,7-tetrahydrothieno[2,3c~pyridine-6-carboxylic acid residue, in place of the (6R)-4,5,6,7-tetrahydrofuro[2,3c~pyridine-6-carboxylic acid residue, in place of the (6R)-4,5,6,7-tetrahydrofuro[3,4c~pyridine-6-carboxylic acid residue, in place of the (6R)-4,5,6,7-tetrahydrofuro[3,2c~pyridine-6--carboxylic acid residue, in place of the (6R)-4,5,6,7-tetrahydro-3Hpyrrolo[2,3- clpyridine-6-carboxylic acid residue, in place of the (6R)-4,5,6,7-tetrahydro-2Hpyrrolo[3,4- c~pyridine-6-carboxylic acid residue, in place of the (6R)-4,5,6,7-tetrahydro-lHpyrrolo[3,2- c]pyridi-ne-6-carboxylic acid residue, in place of the (6R)-2-methyl-4,5,6,7-tetrahydro- 2H- pyrrolo[3 pyridine-6-carboxylic acid residue, in place~ of the (6R)-l-methyl-4,5,6,7-tetrahydro 1H- pyrrolo pyridine-6-carboxylic acid residue, in place of the (6R)-3-methyl-4,5,6,7-tetrahydro- H- pyrrolo pyridine-6-carboxylic acid residue, Ilm] tpp [3m] tpp 11 (3m] tip [im] tip 7] tna 71 tna 7] tna tna epc Gbu Example 1: in place of the (6R)-3-methyl-4,5,6,7-tetrahydro- 3H- imidazo pyridine-6-carboxylic acid residue, in place of the (6R)-l-methyl-4,5,6,7-tetrahydrolH- imidazo[4,5-clpyridine-6-carboxylic acid residue, in place of the (6R)-5,6,7,8-tetrahydro-l,7naphthyridine-6-carboxylic acid residue, in place of the (6R)-5,6,7,8-tetrahydro-2,7naphthyridine-6-carboxylic acid residue, in place of the (1R)-5,6,7,8-tetrahydro-3,7naphthyridine-6-carboxylic acid residue, in place of the (6R'-5,6,7,8-tetrahydro-4,7naphthyridine-6-carboxylic acid residue, in place of the 5-ethoxypiperidine-2-carboxylic acid resiclue, in place of the 2-amino-4-guanidinobutyric acid residue.

Gba-Arg-dhPro-Hyp-Gly-Thi-Ser-tic-Oic- Arg-OH hexafluoroacetate The compound of Example 1 is synthesized substituted with 0.33 mmol/g of Fmoc-Arging to the following repetitive protocol: Operation no. Function Solvent/ Reactant 1 washing DMF 2 deprotection 20% piperidine/DMF 3 deprotection 20% piperidine/DMF 4 washing DMF washing dichloromethane 6 coupling activated protected amino acid f rom 2 g of a resin (Pmc)-OH and accord- Repetition/ time 2 x 2 min 1 x 5 min 1 x 15 min 3 X 3 X 2 rmin 2 min 1 x 90 min 12 7 washing DMF 3 x 2 min 8 washing isopropyl 3 x 2 min alcohol 9 washing dichloro- 3 x 2 min methane Each of these operations, performed in 30 ml of solvent with agitation at room temperature, is followed by filtration through a glass sinter incorporated in the glass cell (reactor) in which the synthesis progresses. The filter retains the resin to which the growing peptide chain is attached.

The chosen protected amino acids were introduced in the following order: Fmoc-Oic-OH, Fmoc-tic-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thi-OH, Fmoc-Gly-OH, Fmoc-Hyp(tBU)-OH, Fmoc-dhPro-OH and Fmoc-Arg(Pmc)-OH.

The activation for the purpose of coupling (operation 6) is obtained in each cycle by dissolving 4 equivalents (2.64 mmol) S of the protected amino acid with 360 mg of HOBt in 30 ml of DMF, and then, after 30 minutes at room temperature, by adding *0 0 618 mg of DCC. This solution is then introduced immediately into the reaction cell with 10 ml of dichloromethane.

At the end of the eight cycles corresponding to the sequential attachment of eight amino acids, and with C-terminal arginine, a nonapeptide protected on its side chains and attached to the resin has thereby been obtained. After treatment under conditions identical to those described for the previous cycles with 4-guanidinobenzoic acid, the resin is then treated with a mixture of trifluoroacetic acid (18 ml), dichloromethane (1 ml) and anisole (1 ml) for 90 minutes at room temperature. The filtrate and the solvents used for washing the resin (3 x 20 ml Of dichloromethane) are combined and evaporated to dryness. The product is suspended in ether, filtered off and dried, then purified by preparative HPLC on a

C

18 column (internal diameter: 47 mm, length: 300 mm) and lyophilized.

Analysis of the product obtained is carried out after decomposition of the latter into amino acids by hydrolysis in 13 6N hydrochloric acid for 18 hours at 110 0 C, and quantitative assay of the amino acids obtained by HPLC. This analysis complies with the standards normally required.

Analysis Arg Hyp Ser Gly dhPro Oic tic Calculated 2 1 1 1 1 1 1 Found 1.99 0.94 0.93 1.1 1.1 0.96 1.00 Mass spectrum (FAB): MH+, m/z 1307 The examples which follow were prepared using the procedure described in Example 1.

Example 2: Gba-Arg-Pro-Hyp-Gly-Thi-Ser-[3,2-c]tpp-Oic-Arg trifluoroacetate The resin substituted with Fmoc-Oic-Arg(Pmc)-OH is condensed after deprotection using piperidine in the same manner as in Example 1 with Fmoc-tpp-OH. This intermediate is obtained from Thi-OH in three steps: thienylalanine hydrochloride is obtained in a methanol solution saturated with HC1; cyclization of the thienylalanine hydrochloride in a 209."% solution of HCHO in water, with stirring for 2 h at 110 0

C

followed by 15 h at 60 0

C;

protection at the Na position with the fluorenylmethyloxycarbonyl group (Fmoc).

Analysis Arg Hyp Ser Gly Pro Oic Thi 23: calculated 2 1 1 1 1 1 1 found 2.09 1.01 0.97 0.99 1.01 1.000.96 Mass spectrum (FAB): MH+, m/z 1315 Example 3: arg-Arg-Pro-Hyp-Gly-Thi-Ser-[3,2-c]ttp-Oic-Arg trifluoroacetate Analysis Arg Hyp Ser Gly Pro Oic Thi [3,2-c]ttp calculated 3 1 1 1 1 1 1 1 found 2.79 1.02 1.00 1.05 1.06 1.03 0.99 1.04 14 Mass spectrum (FAB): MH+, m/z 1309 Example 4: Gba-Arg-Pro-Hyp-Gly-Thi-Ser-tic-Oic-Dpr-OH Ana:lysis Arg Hyp Ser Gly Pro Qic tic Thi calculated 1 1 1 1 1 1 1 1 found 0.99 1.07 0.95 0.97 1.03 1.08 0.92 1.00 Mass spectrum (FAB): MH+, m/z 1239 Example 5: Gba-Arg-Pro-Hyp-Gly-Thi-Ser-tic-oic-Gpr-OH Analysis Arg Hyp Tic Oic Pro Gly Set calculated 1 1 1 1 1 1 1 found 0.97 0.93 1.02 1.00 1.06 1.03 0.99 Mass spectrum (FAB): MH+, m/z 1281 Example 6: Gba-Dbu-Pro-Hyp-Gly-Thi-Ser-tic-Oic-Dbu-OH .Analysis Dbu Oic Tic Ser Gly Hyp Pro calculated 2 1 1 1 1 1 1 .:.found 2.03 1.01 0.99 0.91 1.07 1.02 1.03 Mass spectrum (FAB): MH+, m/z 1196 .::Example 7: Gba-Arg-Pro-Hyp-Gly-Thi-Ser-[3,4-clttp-Oic- Arg-OH Example Gba-Arg-Pro-Hyp-Gly-Thi.-Ser-[2,3-c]ttp-Oic- Arg-OH Example 9: Gba-Arg-Pro-Hyp-Gly-Thi-Ser-[12,3-cltfp-Oic- Arg-OH Example 10: Example 11: Example 12: Example 13: Example 14: Gba-Arg-Pro-Hyp-Gly-Thi-Ser- 3,4-c] tfp-Oic- Gba-Arg-Pro-Hyp-Gly-Thi-Ser- [3 tfp-Oic- Gba-Arg-Pro-Hyp-Gly-Thi-Ser- 12 tpp-Oic- Gba-Arg-Pro-Hyp-Gly-Thi-Ser- 13, 4-c] tpp-Oic- Gba-Arg-Pro-Hyp-Gly-Thi-Ser- 13, 2-cl tpp-Oic- Ar g-OH Arg-OH Arg-OH Arg-OH Arg-OH 15

S

S.

S

*5 S *5 S S

S

SSSS**

1~S S S Example 15: Example 16: Example 17: Example 18: Example 19: Example 20: Exampleft 2 1: Example 22: Example 23: Example 24: Example 251: Example 26: Example 27: Example 28: Example 29: Example 30: Exampl1e 31: Example 32: Example 33: Gba-Arg-Pro-Hyp-Gly-Thi-Ser- [3m] tpp-Oic'-Arg-OH Gba-Arg-Pro-Hyp-Gly-Thi-Ser- [2m] Lpp-Oic-Arg-OH Gba-Arg-Pro-Hyp--Gly-Thi-Ser- [lm] tpp-Oic-Arg-OH Gba-Arg-Pro-Hyp-Gly-Thi-Ser- [3m] tip-Oic-Arg-OH Gba-Arg-Pro-Hyp-Gly-Thi-Ser- [lm] tip-Oic-Arg-OH Gba-Arg-Pro-Hyp-Giy-.Thi-Ser-[l,7]tna-Oic-Arg-

OH

Gba-Arg-Pro-Hyp-Gly-Thi-Ser- II2,7 ]tna-Oic-Arg- OH Gba-Arg-Pro-Hyp-Gly-Thi-ser-[3,7]tna-Oic-Arg-

OH

Gba-Arg-Pro-Hyp-Gly-Thi-Ser-[t4,7 ]tna-Oic-Arg- OH Gba-Arg-Pro-Hyp-Gly-Thi-Ser-epc-Oic-Arg-OH arg-Arg-Pro-Hyp-Gly-Thi-Ser-[3,2-cltfp-oic- Arg-OH arg-Arg-Pro-Hyp-Gly-Thi-Ser- F3,4-c]ltpp-Oic- Arg-OH arg-A-a,-Pro-Hyp-Gly-Thi-Ser- II2,3-c] tpp-Oic- Arg-OH arg-Arg-Pro-Hyp-Gly-Thi-Ser-[3m] tpp-Oic-Arg-OH arg-Arg-Pro-Hyp-Gly-Thi-Ser- [2m] tpp-Oic-Arg-OH arg-Arg-Pro-Hyp-Gly-Thi-Ser-f lm] Lip-Oic-Arg-OH arg-Arg-Pro-Hyp-Gly-Thi-Ser-(3,4-clttp-Oic- Arg-OH arg-1\rg-Pro-Hyp-Gly-Thi-Ser- ttp-Oic- Arg-OH Gba-Arg-Pro-Hyp-Gly-Thi-Ser- ttp-Oic- Gpr-OH 16 9 9 *9 9* 9.

9 *9~9 9* S 9

S

9 9* 9 9 0S59 9 Example 34: Gba-Gpr-Pro-Hyp-Gly-Thi-Ser-13,2-clttp-aic- Gpr-OH Example 35: Gba-Dbu-Pro-Hyp-Gly-Thi-Ser-[3,2-c]ttp-Oic- Gpr-OH Example 36: Gba-Gbu-Pro-Hyp-Gly-Thi-Ser-1~3,2-c] ttp-Oic- Gpr-OH Example 37: arg-Arg-Pro-Hyp-Gly-Phe-Icr-(3,2-clttp-Oic- Arg-OH Example 38: a rg-Arg-Pro-Hyp-Gly-Thi-Sier-Mephe-Oi c-Ar g-OH tetrafluoroacezate Analysis Arg Pro Hyp Ser Gly calculated 3 1 1 1 1 found 3.05 1.05 1.00 0.98 0.90 Mass spectrum (FAB): MH+, m/z =1307 Example 39: Gba-Arg-Pro-Hyp-GlyT'CH 2 S]Phe-Ser-tic-Oic-Arg-OH pentafluoroacetate Analysis Arg Hyp Ser Pro tic Qic calculated 2 1 1 1 1 found 2.14 0.2 0.93 0.93 1.00 0.98 Mass spectrum (FAB): MH+, m/z 1306 Example 40:. Gba-Arg-Pro-Hyp-Gly'[CH 2 S]phe-Ser-tic-Oic-Arg-OH pentafluoroacetate Example 41: Gba-Dpr-Pro-Hyp--Gly-Thi-Ser- tic-Qic-Dbu-OH trifluoroacetate Analysis Dbu OZIC, tic Ser Gly Hyp Pro Dpr calculated 1 1 1 1 1 1 1 1.

f ound 0.98 1.08 0.98 0.94 1.02 0.94 0.98 1.08 Mass spectrum (FAB): MH+, m/z =1183 Example 42 Gba-Arg-Pro-Hyp--Sar-Thi-Ser-t ic-Oic-Arg-OH hexafluoroacetate 17 Analysis Arg Hyp Ser Sar Pro Oic tic calculated 2 1 1 1 1 1 1 found 2.09 0.95 1.02 0.98 0.97 0.95 1.04 Mass spectrum (FAB): MH+, m/z 1323 Example 43: Gba-Arg-Pro-Hyp-Aoc-tic-Oi c-Arg-OH trifluoroacetate Analysis Arg Hyp tic Oic Pro calculated 2 1 1 1 1 found 1.81 1.04 1.03 1.07 1.05 Mass spectrum (FAB): MH+, m/z 1153 PHARMACOLOGICAL STUDY OF THE COMPOUNDS OF THE INVENTION Example 44: In vitro measurement of the effect of the compounds of the invention with respect to the bradykinin-induced contractile response of the smooth muscle of rabbit jugular vein This study was carried out according to the protocol described by D. REGOLI and J. BARABE (B 2 Receptor Pharmacology of Bradykinin and related kinins, Pharm. Rev., 32, 1, 1-46, 1980). Rings of jugular vein are suspended in organ chambers (40 ml) containing KREBS solution, which is thermostated at 37 0 C and oxygenated continuously by bubbling 95% 02/5% CO 2 through it. The isometric force developed by the rings is measured and recorded. The baseline tension is approximately 2 g. The troughs receive the concentrations of the test compound of the invention (10-8M to 10-5M), while a control trough receives the solvent. A curve of concentration versus response to bradykinin is plotted from 10-10M to 10-SM in half-logarithm steps. The potency of the bradykinin antagonists is measured from the log dose-effect curves, enabling the pA2 (negative logarithm of the molar concentration of the compound of the invention which necessitates a doubling of the bradykinin concentration in order to obtain the same effect) values to be determined. In this test, the compounds of the invention have 18 a pA2 of between 9 and 10. More especially, the pA2 of the compound of Example 2 is equal to 9.1 0.1, that of Example 3 is equal to 9.4 0.1 and that of Example 5 is equal to 9.3 0.1.

Example 45: In vivo measurement of the anti- inflammatory activity of the compounds of the invention The anti-inflammatory activity of the compounds of the invention was measured using the model of edema of the rat's foot induced by injection of carrageenan (WINTER G. A. et al., Proc. Soc. Exp. Biol. Med., 3, 544- 547, 1960).

This study is performed on male rats weighing 180-210 g in groups of 8. The compounds of the invention are administered I.V. at time 0 immediately preceding the subcutaneous injection rf carrageenan into the sole of the rat's right hind S" foot (type IV lambda carrageenan, Sigma, 1% solution, volume S injected 0.1 ml). The volume of the foot at different timepoints of the experiment is measured by plethysmography. The inhibition of the edema at time 3 hours is calculated relative .to a control group which has received the vehicle, and expressed as a percentage inhibition.

Under these conditions, the compounds of Examples 2 and 3 produce a 36 inhibition at time 1 hour at a dose of 0.1 mg/kg and the compound of Example 5 produces a 55% inhibition at time 1 hour at the same dose.

Example 46: Pharmaceutical composition Ointment containing 5 pg/ml of the peptide of Example 1 Gba-Arg-dhPro-Hyp-Gly-Thi-Ser-tic-Oic-Arg-OH 50 mg Polyethylene glycol Q.S 100 ml Example 47: Solution for injection Gba-Arg-dhPro-Hyp-Gly-Thi-Ser-tic-Oic-Arg-OH 0.5 mg Distilled water for injections Q.S. 25 ml

Claims

1.A compound of formula \Al-A23-A-A4-A5-A6-A 7 -A8-A 9 -Y (I) in which: X a f-u substituent of the terminal amino group of the peptide of formula wh--h ma-; be l -4le:e= represents: a hydrogen atom, a 4-guanidinobenzoyl radical (Gba), unsubstituted or substituted on the phenyl ring with one or more halogen atoms or hydroxyl, mercapto, linear or branched (C 1 -Cs) alkyl, linear or branched (C 1 -Cs) alkylthio or linear or branched (C 1 -Cs) alkoxy radicals, a linear or branched (C 1 -Cs) alkyl radical, a (C 6 -C 12 aryl radical, a linear or branched (C 1 -C 6 alkoxycarbonyl radical, a linear or branched (Cl-C 6 alkylsulfonyl radical, a 9-fluorenylmethyloxycarbonyl radical, a linear or branched (C 1 -C 6 guanidinoacyl radical, unsubstituted or substituted with an amino radical, or a radical of formula /NH2 H2N-(CH2)n-CH in which n is between 1 and 6, A, and Ag, which may be identical or differen present: a bond, teie of formula I I 19a a substituent of the terminal amino group of the peptide of formula hydrogen, A 1 represents: a residue of formula e S S S S .5 S 55 S S S S 55 S. S ~5 S S 20 R-(CH2)n-CH \C in which n represents an integer between 1 and 6 and R, an amino or guanidino radical, -or any basic amino acid, unsubstituted or substituted on its side chain, A 2 and A 3 which may be identical or different, represent: ai -a proline (Pro), hydroxyproline (Hyp), 2-azabi- cycloF.2.2.2loctane-3-carbonyl (Aba), 2-azabi- cyclo(i2.2.llheptane-3-carbony. (Abh), homoproline (hPro), 3,4-dehydroproline (dhPro), sarcosine (Sar), N-methylphenylglycine (NMePgl), azetidine- 2-carbonyl (Azt) or alanine (Ala) residue, A 4 represents A za glycine (Gly) or sarcosine (Sar) residue or a residue of formula: -NH- (CH 2 m-CO3 in which m is an integer between 2 and 8, A 5 represents h 2-n ar a phenylalanine (Phe), f3-(2- thienyl)alanine (Thi), J-cyclohexylalanine (Cha), 1D- naphthylalanine (Nal), 1-furylalanine (Fur), J3- pyranylalanine (Pra) or J-pyridylalanine (Pyr) residue, A 6 represents Aba any amino acid residue, A 7 any one of the following residues: N N N N N NN 0j Co/o 0 CO o CO B C D E F i A 21 H N N CO CH 3 N N Co N N Co P N -cH 3 N Co K N 8N Co Q CH 3 NI N CO L N Co R CH 3 N 7 Co 0 M N N Co (R represents a radical) linear: or branched (Cl-C 6 represents a methyiphenylalanine residue (Me.?he) tetrahydroisoquinoline-3-carbonyl residue (Tic condition that, in this case, X represents guanidinobenzoyl radical and X' a hydrogen atom, A 8 represents =e4 an octahydroindole-2-carbonyl (Oic), p ro 1i n e (Pro), hydroxyproline Hyp, 2 -a za bicycloj2.2.2]octane-3-carbonyl (Abo), 2-azabi- cyclo[2.2.1]heptane-3-carbonyl (Abh), homoproline (hPro), 3,4-dehydroproline (dhPro), sarcosine (Sar), N- methyiphenyiglycine (NMePgl), azetidine-2--carbonyl (Azt) or alanine (Ala) residue or any one of the residues A to R defined for A 7 Y, a substituent of the terminal carbonyl qgro-p-of--EHi Alpeptide of formula rejr~t a hydroxyl or linear or "/ranched(a .6 alkoxy radical or an amino radical 21 a A 9 a residue c i formula /NH- Rl-(CH2)n-CH Co in which n represents an integer between I and 6 and R, an amino or guanidino radical, or any basic amino acid, unsubstituted or substituted on its side chain, Y, a substituent of the terminal carbonyl group of the peptide of formula represents a hycdroxyl or linear or branched (01-06) alkoxy radical or an amino radical 22 (unsubstituted or substituted with one or two linear or branched (CI-C 6 alkyl radicals or phenyl radicals it being possible for each of the alkyl or phenyl radicals itself to be substituted with an amino, guanidino or ureido radical), or alternatively the carbonyl-terminal group of the peptide of formula is reduced to the corresponding alcohol (ol), and its addition salts with a pharmaceutically acceptable acid or base, on the understanding that: when X represents a p-guanidinobenzoyl radical, X a hydrogen atom, Al an arginine residue (Arg), A 2 a proline residue (Pro), A 3 a hydroxyproline residue (Hyp), A 4 a glycine residue (Gly), A 5 a -(2-thienyl)alanine residue *i (Thi), A 6 a serine residue (Ser), As an octahydroindole-2- carbonyl residue (Oic), Ag an arginine residue (Arg) and Y a hydroxyl radical, then A 7 is other than a tetrahydroisoquinoline-3-carbonyl residue (Tic) of D or L configuration with respect to its a carbon, each peptide bond -CO-NH- of the peptide of the formula can be optionally replaced by a pseudopeptide bond chosen, from -CH 2 -NH, -CH 2 -CH 2 -CH 2 -S0 2 -NH-CO- or -CH=CH-, each amino acid of the peptide sequence being optically pure and the a carbon of each amino acid having the D or L configuration.

2.A compound of formula as claimed in claim 1, such that X represents a 4-guanidinobenzoyl radical (Gba). .a n a-d %E~ A 23

3.A compoundi of formula as claimed in claim A 7 represents any one of the following residues: N N N N 7/7 Co 0 CO CO N /N 0C D CH 3 N N Co N N Co P 1, such that N Co F CH 3 3 ri N CO L a. a a a a. a. a a a a a. 9. a a a a *aa* a~ a. a a a a a aa.~ CH 3 CO RO N Co CH 3 N /CO N N 8N /Co 0 N CO Q \N N CO R (R represents a linear or branched (Cl-C 6 radical)

4.A compound of formula JI) as claimed in claim 3, such that X represents a 4-guanidinobenzoyl radical (Gba).

5.A compound of formula as claimed in claim 3, such that X represents an arginyl radical,:an M 4 In I 24

6.A process for preparing a compound of formula as claimed in claim 1, which compound may be obtained by sequential solid-phase synthesis from protected amino acids, by synthesis from peptide fragments in solution, or, where appropriate, by combination of these two techniques, and, if so desired, by introduction, according to standard methods of organic chemistry, of a pseudopeptide bond at any point in time during the synthesis of the peptide sequence, which compound of formula is converted, if necessary, to its addition salts with a pharmaceutically acceptable acid or base.

7.A pharmaceutical composition containing as active principle at least one compound as claimed in any one of claims 1 to alone or in combination with one or more pharmaceutically acceptable excipients or vehicles.

8.The pharmaceutical composition as claimed in claim 7, containing at least one active principle as claimed in any one of claims 1 to 6, which is useful as a bradykinin antagonist in the treatment of traumas of diverse origins, inflammatory disorders, pain, shock states, allergic disorders and insufficiency of spermatozoal mobility. DATED this 16th day of June 1993. "ADIR ET COMPAGNIE S• WATERMARK PATENT TRADEMARK ATTORNEYS "THE ATRIUM" 290 BURWOOD 1OAD HAWTHORN. VIC. 3122. ABSTRACT NEW PEPTIDE COMPOUNDS 14AVING BRADYKININ-ANTAGONISTACTIVITY, PROCESS FOR THEIR PREPARATION AND THE PHARMACEUTICAL COMPOSITIONS WHICH CONTAIN THEM ADIR ET COMPAGNIE 1. RUE CARLE HERBERT 92415 COURBEVOIE CEDEX S. I I I. I I 1* Compounds of formula x \/A1 -A2-A3-A)4-A5-A6-A 7 -A8-A 9 -Y in which X, X A 2 A 3 A 4 A 5 A 6 A 7 As, Ag and Y are as defined in the description. a.

9 4 III. Medicinal products.