LT3872B - Novel peptides and pharmaceutical compositions containing them - Google Patents

Abstract

Formula I peptides:A-B-C-E-F-K-(D)-Tic-G-M-F'-Iwhere A is hydrogen, alkyl, alkanoyl, alkoxycarbonyl, alkylsulfonyl, cycloalkyl, aryl, arylsulfonyl, heteroaryl or amino acid that in case of need may contain substitutes,B is basic amino acid,C is di- or tripeptide,E is remnant aliphatic of alicyclically-aliphatic acid,F is amino acid or a direct combination, independently of each other in case of need containing substitutes in a side chain,G is amino acid,F' has the same meanings as the F or may be -NH-(CH2)2-8<-> or a direct combine,I is -OH, -NH2 or -NHC2H5 andK is a remnant or a direct combination of -NH-(CH2)1-4-CO- distinguished by an antagonistic effect to the bradykinin. Their therapy effect covers various pathogenic states that are stimulated, caused and sustained by the bradykinin and bradykinin allied peptides. Formula I peptides are produced by known peptide synthesis methods.

Description

The present invention relates to novel peptides having bradykinin antagonistic activity and to pharmaceutical compositions comprising these compounds.

Antagonistic bradykinin peptides are described in WO 86/07263; in these peptides, among other things, the 7-position L-Pro of the bradykinin peptide hormone or other bradykinin analog is replaced by a D-amino acid such as D-Phe, D-Thia, D-Pal, CDF, D-Nal, MDY, D -Phg, D-His, D-Trp, D-Tyr, D-hPhe, D-Val, D-Ala, D-His, D-Ile, D-Leu, and DOMT.

The main object of the invention is to find new effective peptides which have antagonistic effect on bradykinin.

This problem is solved by peptides of formula I:

A-B-C-E-F-K - (D) -You - G-M-F'-I (I) in which:

A a) is hydrogen, (C 1 -C 6) -alkyl, (C 1 -C 8) -alkanoyl, (C 1 -C 8) -alkoxycarbonyl or (C 1 -C 8) -alkylsulfonyl, optionally one, two or three hydrogen atoms may be substituted by one, two or three identical or different residues, such as carboxyl, amino, (C₁-C₄) alkyl, (C, -C ₄₎ -alkilaminogrupė, hydroxyl, _{(C] a} -q ) -alkoxy LT 3872 group B, halogen, di- (C 1 -C 4) -alkylamino, carbamoyl, sulfamoyl, (C 1 -C 4) -alkoxycarbonyl, (C 1 -C 4) -aryl and (C 1 - 4). C ._) 14 b 1Z blz

-aryl- (C 1 -C 4) -alkyl; or wherein optionally one hydrogen atom, in this case, is substituted with one residue such as (C 1 -C 6) cycloalkyl, (C 1 -C ₄ ) alkylsulfonyl, (C ₁ -C ₄ ) alkylsulfinyl, (C ₈ ) -C 1-4 -aryl- (C 1-4 -C ₆ ) -alkylsulfonyl, (C 8 -C ₁₂ ) -aryl- (C 1 -C 4 -C 1-4 alkylsulfinyl), (C 8 -C 4 -C ₁ ) aryl group, ( -C) -heteroaryl and (C 1 -C 8) -heteroaryloxy, and one or two hydrogen atoms replaced by one or two same or different residues such as carboxyl, amino, (C 1 -C 4) -alkyl, hydroxy, ( C 1 -C ₄ alkoxy, halogen, di- (C 1 -C ₄ ) alkylamino, carbamoyl, sulfamoyl, (C ₁ -C ₄ ) - alkoxycarbo-

a ₂ ) is (C 1 -C 8) -cycloalkyl, carbamoyl, which may optionally be substituted on the nitrogen atom with (C 1 -C 8) -alkyl or (C 8 -C ₁₂ ) -aryl; (C 8 -C 6) -aryl, (C 2 -C 4) -aryl, (C 8 -C 4) -arylsulfonyl, (C 1 -C 6) -heteroaryl or (C 1 -C 4) -heteroaryl;

besides, a ^) and a ^} p. labeled residues such as aryl, heteroaryl, aroyl, arylsulfonyl and heteroaryl may be optionally substituted with 1, 2, 3 or 4 substituents, such as:

carboxyl, amino, nitro, (C 1 -C 4) -alkylamino, hydroxy, (C 1 -C 4) -alkyl, (C 1 -C 4 -C 4) -alkoxy, halogen, cyano, di- (C 1 - 4) C 1-4 alkylamino, carbamoyl, sulfamoyl and (C 1-4 C) alkoxycarbonyl, or) is a residue of formula II:

N - CH

C

II (II) wherein A is as defined in (a ^) and a £);

R is hydrogen or methyl;

R is hydrogen or (C 1 -C 4) -alkyl, preferably (C 8 -C 1 -C 4) -alkyl, which may optionally be substituted with one substituent such as amino, substituted amino, hydroxyl, carboxyl, carbamoyl, guanidine group, substituted guanidine group, ureido group, mercaptogroup, methyl mercaptogroup, phenyl, 4-chlorophenyl, 4-fluorophenyl, 4-nitrophenyl, 4-methoxyphenyl, 4-hydroxyphenyl, phthalimido group, 4-imiaazolyl, 3-indolyl, 2- thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl or cyclohexyl, wherein the substituted amino group is -NH-A and the substituted guanidine group is -NH-C (NH) -NH-A, wherein A is as defined a ^) or a ^);

B is a basic amino acid of the L- or D-configuration which may be substituted in the side chain;

C is a grouping of formulas (llla) and (lilb):

G '- G' - Gly G '- NH - (CH 2) - CO ^J 2 n (IIIa) (IHb) wherein G', independently of one another, is a group of formula IV:

- N - CH - C ι i. ii (IV),

R 'where

5 ....

R and R together with the atoms to which they are attached form a heterocyclic mono-, bi- or tricyclic system with 2-15 C atoms and n = 2-8;

E is a neutral, acidic or basic, aliphatic or allylic cyclic-aliphatic amino acid residue;

F, independently of one another, is a neutral, acidic or basic, aliphatic or aromatic amino acid residue which may be substituted in the side chain; or is a direct connection;

D-Tic is a residue of formula V:

G is the same as G ', whose values are given above, or is a direct bond;

F 'is the same as F, is a -NHCCi ^) ^ - residue, where n = = 2-8, or, if G is not a direct bond, may be a direct bond;

I is -OH, -NH ₂ or -NHC ₂ H ₅ ;

K is a residue of -NH- (CH_) -CO-, where x = 1-4, or is simply

Z. X no connection, o

M has the same meanings as F; and physiologically acceptable salts of these compounds.

Unless otherwise indicated, truncation of an amino acid without reference to a stereo tag denotes an L-shaped amino acid residue (See Schrodes, Lubke, The Peptides, Vol. 1, New York, 1965, XXII-XXV; Houben-Weyl, Methoden der Organische). Chemie, Bd.XV / I -2, Studgard, 1974), for example:

Aad, Abu, yAbu, ABz, 2ABz, £ Aca, Ach, Acp, Adpd, Ahb, Aib, pAib, Ala,) 3Ala, δ Ala, Alg, Ali, Ama, Amt, Ape, Apm, Apr, Arg, Asn , Asp, Asu, Aze, Azi, Bai, Bph, Can, Cit, Cys, Cyta, Daad, Dab, Dadd, Dap, Dapm, Dasu, Djen, Dpa, Dtc, Fel, Gln, Glu, Gly, Guv, i

hAla, hArg, hCys, hGln, hGlu, His, hlle, hLeu, hLys, hMet, hPhe, hPro, hSer, hThr ,, hTrp, hTyr, Hyl, Hyp, 3Hyp, Ile, Ise, Iva, Kyn, Lant, Lcn , Leu, Lsg, Lys, ^> Lys, ALys, Met, Mim, Min, nArg, Nle, Nva, Oly, Orn, Pan, Pec, Pen, Phe, Phg, Pic, Pro, zlPro, Pse, Pya Pyr, Pza, Qin, Ros, Sar, Sec, Setu, Ser, Thi, ^> Thi, Thr, Thy, Thx Tia, Tie, Tly, Trp, Trta, Tyr, Vai.

As residues of the heterocyclic system indicated in the description of formula IV, acceptable residues of the heterocycle from the following group are acceptable:

pyrrolidine-2-carbonic acid, piperidine-2-carbonic acid;

1,3,3,4-tetrahydroisoquinoline-3-carbonic acid, decahydroisoquinoline-3-carbonic acid, octahydroindole-2-carbonic acid, decahydroquinoline-2-carbonic acid, octahydrocyclopenta (b) pyrrole-2-carbonic acid,

2-aza-bicyclo (2.2.2) -octane-3-carbonic acid,

2-azabicyclo (2.2.1) -heptane-3-carbonic acid, 2-azabicyclo (3.1.0) -hexane-3-carbonic acid,

2-azaspiro- (4.4) -nonan-3-carbonic acid,

2-Azaspiro- (4 .:) - decane-3-carbonic acid, spiro-bicyclo (2.2.1) -heptane / -2,3-pyrrolidine-5-carbonic acid, spiro-bicyclo (2.2. 2) octane / -2,3-pyrrolidine-5-carbonic acid,

9th

2-Azatricyclo (4.3.0.1 ') decane-3-carbonic acid, decahydrocyclohepta (b) pyrrole-2-carbonic acid, decahydrocycloocta (b) pyrrole-2-carbonic acid, octahydrocyclopenta (c) pyrrole-2-carbonic acid, octahydroisoindole -l-carbonic acid,

2,3,3a, 4,6a-hexahydrocyclopenta (b) pyrrole-2-carbonic acid 2,3,3a, 4,5,7a-hexahydroindole-2-carbonic acid, tetrahydrothiazole-4-carbonic acid, isoxazolidine-3- carbonic acid, pyrazolidine-3-carbonic acid, hydroxyproline-2-carbonic acid, which may be optionally substituted.

i t

i

The heterocycles which are based on the above-mentioned residues are known, for example, from the following patents: US-A 4 344 949, US-A

No. 374,347, US-A 4 350 704, EP-A 29,488, EP-A 31 741, EP-A 46 953 -A 49 605, EP-A 49,658, EP-A 50,800, EP-A 51 020, EP-A 52 870, -A 79 022, EP-A 84,164, EP-A 89,637, EP-A 90 341, EP-A 90 362, -A 105 102, EP-A . 109,020 , EP-A iii 873, EP-A 271,865 and EP-A 4,682.

Unless otherwise stated in the particular case, alkyl may be linear or branched. The same is true for alkyl moieties such as alkoxy, aralkyl or almoyl.

(C 8 -C 12) -aryl preferably represents phenyl, naphthyl or bisnyl. The same can be said for the residues of its derivatives, a torus as an aryloxy group, an aralkyl or an aroyl.

Halogen means fluorine, chlorine, bromine or iodine, preferably chlorine.

Acceptable focal salts include, in particular, alkali or alkaline earth metal salts, physiologically acceptable amine salts, and inorganic or organic acids such as, for example, HCl, IBr, H ₂ SO ₄ , ΡΟ ^ ΡΟ ^, maleic, fumaric, citric, in wine, acetic acid and yeast.

Most preferred are peptides of formula I wherein:

B is Arg, Lys, Orn, 2,4-diaminobutyryl or a residue of L-homoarginine, and, where appropriate, the side chain amino or guanidine group may be replaced by the group A as defined in a) and a) ₂ ). ;

E is the L- or D-configuration or alicikliškaialifatinės aliphatic amino acid residue which has a side chain 1-14 C-ato tions such as alanine, serine, threonine, 0- (C₁-C₆) -alkyl or 0- (C _r - C "-) - aryl-blocked serine or threonine, valine, and 1U norvaline, leucine, isoleucine, no rleucine, neppeptylglycine, tert-butylglycine or (C 8 -C 8) -cycloalkyl- (C 1 -C 8) -alkylglycine;

F 'is a residue of a basic amino acid of the L- or D-configuration, such as Arg or Lys, wherein the guanidine group or amino group of the side chain may be replaced by A as described in a ^) and ag), or is -NH-iCHg) ^. -, where n = 2-8, a residue;

K is a residue of -NH- (CHg) -CO-, where x = 2-4, or a direct bond:

Particularly suitable are peptides of the formula I in which:

B is Arg, Orn or Lys, wherein guanidine, or the corresponding amino groups in the side chain, are unsubstituted or may be substituted such as (C 1 -C 8) -alkanoyl,

-aryl, (C 8 -C 8) -heteroaryl, (C 1 -C 6) -alkylsulfonyl or (C 4 -C 10) -arylsulfonyl, wherein the aryl, heteroaryl, aroyl, aryl ± z sulfonyl and heteroaryl moieties as described in ) may have 1, 2, 3 or 4 substituents, if necessary, identical or different;

E is leucine, isoleucine, norleucine, tert-butylglycine, serine, threonine or cyclohexylalanine;

K is a direct connection, and. . M is a direct connection.

Particularly suitable are peptides of formula I wherein:

A is hydrogen, (D) - or (L) -H-Arg, (D) - or (L) -H-Lys or (D) - or (L) -H-Orn;

B is Arg, Orn or Lys, where the guanidino side chain, or the amine may be substituted with a hydrogen, ^C] _ ~ ^c g) alkanoyl, (C ^ -C ^ g) -aroilas (Cg-Cg) -heteroaroilas , (C 1 -C 6 -C 8) -alkylsulfonyl or (C 8 -C 8 -g) -arylsulfonyl whose aryl, heteroaryl, aroyl, arylsulfonyl and heteroaryl moieties may optionally have 1, 2, 3 or 4 substituents, which may be the same or different. , such as methyl, methoxy and halogen;

C is Pro-Pro-Gly, Hyp-Pro-Gly or Pro-Hyp-Gly;

E is Leu, Ile, Tbg, or Cha;

F is Ser, hSer, Lys, Leu, Vai, Nle, Ile or Thr;

K is a direct bond;

M is a direct bond;

G is a moiety of the heterocyclic system of formula IV, wherein the most preferred moieties are: pyrrolidine-2-carbonic acid, piperidine-2-carbonic acid; tetrahydroisoquinoline-3-carbonic acid; eis and trans -decahydroisoquinoline-3-carbonic acid; cis-endo-, cis-exo-, trans-octahydroindole-2-carbonic acid; cis-endo-, cis-exo, trans-octahydrocyclopentalb) pyrrole-2-carbonic acid or hydroxyproline-2-carbonic acid;

F 'is Arg, o

I is OH.

Particularly suitable examples of formula I peptides include:

H- (D) -Arg-Pro-Hyp-Gly-Leu-Ser- (D) -Tic-Oic-Arg-OH;

H- (D) -Arg-Pro-Hyp-Gly-Cha-Ser- (D) -Tic-Oic-Arg-OH;

H- (D) -Arg-Pro-Hyp-Gly.-Tbg-Ser- (D) -Tic-Oic-Arg-OH.

The invention further relates to a process for the preparation of peptides of formula I, which differs in that

(a) a moiety with a free C-terminal carboxyl group or an activated derivative thereof is exposed to the corresponding moiety with an N-terminal amino group; or

(b) the peptide is synthesized stepwise, followed by the cleavage of one or more protecting groups, if necessary, protected by the compounds of (a) or (b), and converted into the physiologically acceptable salts thereof, where appropriate. .

The peptides of the present invention are obtained by known methods of peptide chemistry (see, e.g., Houben-Weyl, Methcden der Organische Chemie,

15/2 Bd.), Preferably using the solid phase synthesis described, for example, in B. Merrfield, J. Amer. Chem. Soc., 85, 2149 (1963), or B.C. Sheppard, Int. J. Peptide and Protein Res., 21, 118 (1983), or similar methods known in the art. Urethane protecting groups such as, for example, tert-butoxycarbonyl (Boc) or fluorenylmethyloxycarbonyl (Fmoc) protecting groups are used to protect the? -Amino group. If side reactions are to be avoided, or in the case of specific peptide synthesis, the amino acid side chain functional groups are further blocked using appropriate protecting groups (see, e.g., T.W. Greene, Protective Groups in Organic Synthesis); most commonly used:

Arg (Tos), Arg (Mts), Arg (Mtr), Arg (Pmc), Asp (OBzl), Asp (OBut),

Cys (4-MeBzl), Cys (Amc), Cys (SBut), Glu (OBzl), Glu (OBut), His (Tos), His (Fmoc), His (Dnp), His (Trt), Lys (Cl). -Z), Lys (Boc), Met (O), Ser (Bzl), Ser (But), Thr (Bzl), Thr (But), Trp (Mts), Trp (CHO), Tyr (Br-Z) , Tyr (Bzl) or Tyr (But).

Solid phase synthesis begins with the interaction of the terminal carbon atom of the peptide with an amino acid blocked by the corresponding resin. Starting materials of this type may be obtained by coupling a blocked amino acid with a modified polystyrene or polyacrylamide resin of a chloromethyl, oxymethyl, benzhydrylamine (BHA), methylbenzhydrylamine (MBHA) ester to form an ester or amide bond. Resin carrier materials are commercially available. BHA and MBHA resins are commonly used when the peptide synthesized must have a carbamoyl group at its C-terminus. If the peptide is to have a secondary amide group at the terminal v

carbon atom, using a chloromethyl or oxymethyl resin, and resolving the corresponding amines. If, for example, ethylamide is produced, the resin can be cleaved from the resin with ethylamine; the side-chain protecting groups are then separated with other suitable reagents. If the peptide is to retain amino acid side chain protective tert-butyl groups, then the Fmoc group is used to block the amino acid ix-amino group and the synthesis is carried out according to the procedure described, for example, in R.C. Sheppard, J.Chem. Soc. Chem. Commun., 1982, 537, and the arginine guanidine group is protected by protonation with pyridine perchlorate; other amino acid side chain functional groups are blocked by the use of benzylic groups which are subsequently cleaved by catalytic transforhydrogenation (A. Felix et al., J. Org. Chem., 13, 4194 (1978)) or by use of sodium in liquid ammonia (W. Roberts, J. Amer. Chem. Soc., 7 6, 6203 1954).

Once the protecting group which blocks the amino acid group of the resin-linked amino acid is removed, a suitable reagent, such as trifluoroacetic acid in methylene chloride for the Boc protecting group or 20% piperidine in dimethylformamide for the Fmoc protecting group, is gradually coupled. The resulting intermediate peptide resins with N-blocked ends are unblocked using the above reagents before being coupled to the following amino acid derivatives.

The coupling reagent may be any activating reagent used in peptide synthesis (see, e.g., Houben-Weyl, Methoden der Organische Chemie, Bd.15 / 2), for example acetic acid diimides such as N, N ' -diisopropylcarbodiimide, N _T N'-dicyclohexylcarbodiimide or N-ethyl-N '- (3-dimethylaminopropyl) carbodiimide. Coupling can be accomplished directly by coupling the amino acid derivative with an activating reagent and, if necessary, an additive that inhibits racemization such as, for example, 1-oxybenzotriazole (HOBt) (W. Konig, R. Geiger,

Chem. Ber. 103, 708 (1970)) or 3-oxo-4-oxo-3,4-dihydrobenzotriazine (HOObt) (W. Konig, R. Geiger, Chem. Ber. 103, 2054 (1970)), at resin, or can be individually activated by reacting the amino acid derivative by converting it into a symmetric anhydride or HOBt or HOObt ester, and mixing the solution of the activated derivative in a suitable solvent with a peptide resin capable of binding to the derivative.

The coupling or activation of the amino acid derivatives with one of the above activating reagents can be carried out in dimethylformamide, N-methylpyrrolidone or methylene chloride, or in a mixture of the above solvents. Generally, an excess of 1.5 to 4 times the activated amino acid derivative is used. In cases where incomplete coupling occurs, the coupling reaction is repeated without blocking the c? -Amino group of the peptide resin, which is usually required to attach a further amino acid.

The success of the coupling reaction can be conveniently controlled by the ninhydrin reaction as described by E. Kaiser et al. al., Ann. Biochem., 34, 595 (1970). Synthesis can be accomplished automated, for example using an Applied Biosystems Model 430A peptide synthesizer; in this case, either fusion programs provided by the device manufacturer or user-created applications may be used. These latter are used when amino acid derivatives are blocked by Fmoc groups.

The peptides synthesized as described above can be separated from the resin by reagents such as, for example, liquid hydrofluoric acid (preferably for Boc-derived peptides) or trifluoroacetic acid (preferably for Fmoc-derived peptides). These reagents not only cleave the peptide from the resin, but also other protecting groups on the side chains of the amino acid derivatives. In this way, free acid form peptide is obtained using BHA and MBHA resins. Cleavage with hydrofluoric acid or trifluoromethanesulfonic acid yields, in the case of BHA and MBHA resins, an amide peptide. Other methods for obtaining peptide amides are described in EP-A 287 882 and EP-A 322 348. Here, peptides are cleaved from the resin by treatment with moderate acids (e.g. trifluoroacetic acid) commonly used in peptide synthesis; cationic bonding agents such as phenol, cresol, thiocresol, anisole, thioanisole, ethanedithiol, dimethylsulfide, ethylmethylsulfate or the like, e.g. conventional cationic binders in solid-state synthesis, both individually and in mixtures of two or more of these excipients. Trifluoroacetic acid may be used and diluted with other suitable solvents, such as methylene chloride. If it is necessary to preserve the tertiary-butyl or benzyl protecting groups on the peptide side chains, the peptide synthesized on the modified resin carrier is cleaved with 1% trifluoroacetic acid in methylene chloride, for example according to R.C. Sheppard, J. Chem. Soc. Chem. Comm., 1982, 587. Where appropriate, a suitable combination of synthesis and resolving techniques may be used to protect the side chains of the tert-butyl or benzyl groups.

The modified resin carrier described by Sheppard is also used for the synthesis of peptides with a C-terminal amide group or a to-amino group, respectively W-guanidine alkyl. The synthesized peptide having fully blocked side chains is cleaved from the resin and then treated with the appropriate amine, h} -aminoalkylamine or ω-guanidinoalkylamine according to classical synthesis; in addition, other functional groups may be blocked in known ways, if necessary.

Another method for preparing peptides with ω-aminoalkyl group is described in EP-A 264 802.

The peptides of the present invention were synthesized using solid phase synthesis according to two universal tactics for protecting groups.

The synthesis is carried out using an Applied Biosystems Model 430A Automated Peptide Synthesizer using Boc- (respectively Fmoc-) protecting groups to temporarily protect the Gt-amino group.

Using the Boc-protecting group, the synthesis is performed according to the synthesis cycles programmed by the instrument manufacturer.

Synthesis of peptides with a free carboxyl group at the C-terminus is accomplished using 4- (hydroxymethyl) -phenylacetamidomethyl-polystyrene resin functionalized by the appropriate Boc-amino acid Applied Biosystems (R.B. Merrifield, J. Org. Chem., 1978, 43, 2845). MBHA resin from the same company was used to obtain the peptideamide.

N, N'-dicyclohexylcarbodiimide or N, N'-diisopropylcarbodiimide are used as activating reagents. Activation yields symmetric anhydride, HOBt-ester or HOObt-ester in methylene chloride, CH 2 Cl 2 -DMF mixtures or NMP. Used for connection

2-4 equivalents of activated amino acid derivative. In cases where the merger is incomplete, the reaction is repeated.

If a Fmoc-protecting group is used to temporarily <% - protect the amino group, a custom-made program is used to synthesize the Applied Biosystems Model 430A Automated Peptide Synthesizer. The synthesis is carried out using p-benzyloxybenzyl alcohol type resin (S. Wang, J. Amer. Chem. Soc., 1973, 95, 1328), manufactured by Bachem; this resin is esterified with the corresponding amino acid according to known methods (E. Atherton et al., J. Chem. Soc. Chem. Conun., 1981, 336). Activation of the amino acid derivatives to yield the HOBt or HOObt ester is accomplished directly in the amino acid cartridge attached to the device by adding a solution of diisopropylcarbodiimide in DMF to a weighed mixture of the amino acid derivative and HOBt or HOObt. Alternatively, the Fmoc-amino acid-OObt esters prepared as described in EP-A 247 573 may also be used as a separate material. The Fmoc protecting group is cleaved using a 20% piperidine solution in DMF simply in the reaction vessel. The excess of the reactive amino acid derivative used is 1.5-2.5 equivalents. If coupling is incomplete, it is repeated as in the Boc method.

The peptides of the invention (alone or in combination) have an antagonistic effect on Brahaquinine, which can be tested using various models (see Handbook of Exp. Pharmacol., Vol. 25, Springer Ed., 1970, p.53-55), e.g. isolated rat uterus, guinea pig colon, or guinea pig isolated pulmonary artery.

In order to test the peptides of the invention using isolated pulmonary artery, a guinea-pig (Dunkin Hartley) weighing 400-500 g is struck by the occiput, the thoracic cavity is opened and the pulmonary artery is carefully removed. The surrounding tissue is carefully removed and the pulmonary artery is incised spirally at 45 °.

The strips of blood vessels, 2.5 cm long and 3-4 mm wide, were fixed in a 10 ml organic bath filled with Ringer's solution.

Solution composition (mol / l):

Well Cl

154

KC1

CaCl

NaHCO

1.9

2.4

Glucose

5.0

The solution is purged with 95% O _? and 5%. CC> _{2 and} heated to 37 ° C. pH is 7.4; primary load on vascular strips - 1 g Isometric contraction changes are monitored by a lever attachment and an HF modem (knee gauge) (Hugo Sachs firm) and recorded by a compensation recorder (BEG, Goerz Metrawalt SE 460).

After 1 hour for equilibrium, the test begins. After the vascular strips reach maximal 7 * mal sensitivity to 2x10 mol / L bradykinin (bradykinin _g narrows the vascular strips), 5x10 peptides are added.

- 1x10 at a dose of ⁸ mol / l and held for 10 minutes, followed by the addition of bradykinin and a comparable reduction in the effect of bradykinin in control.

1x10 -3 are used to determine the partial-antagonistic effect

- 1x10 mol / l peptide doses.

The values of the &quot; peptides.IC- _n &quot;, as described in the invention, calculated from the dose-effect curves are given in Table 1.

table

Compound ^IC 5i

H- (D) -Arg-Arg-Pro-Hyp-Gly-Leu-Ser- (D) -Tic-Oic-Arg-OH 5,9.10 H- (D) -Arg-Arg-Pro-Hyp-Gly- Cha-Ser- (D) -Tic-Oic-Arg-OH 3.7.10 H- (D) -Arg-Arg-Pro-Hyp-Gly-Tbg-Ser- (D) -Tic-Oic-Arg-OH 6 , 0.10 D-Arg / Hyp ² , Thia ⁵ ' ⁸ , D-Phe ⁷ / BK

H - (D) - Arg-Arg-Hyp-Pro-Gly-Thia-Ser - (D) - Phe-Thia-Arg-OH 6.4.10

The therapeutic use of the peptides of the invention encompasses all pathological states induced, promoted or maintained by bradykinin and related bradykinin peptides. Conditions include injuries such as wounds, burns, as well as rashes, erythema, edema, angina, arthritis, asthma, allergies, rhinitis, shock, inflammation, hypotension, pain, itching and sperm motility disorders.

Therefore, the present invention is also directed to the use of compounds of formula I in medicaments and pharmaceutical preparations containing them.

Pharmaceutical formulations contain an effective amount of the active ingredient of Formula I, either alone or in combination with an inorganic or organic pharmaceutical. carrier material used.

The compounds may be administered enterally, parenterally, for example, subcutaneously, intramuscularly, intravenously, sublingually, subcutaneously, nasally, rectally, vaginally, orally, or by inhalation. The dose of the active substance depends on the type of warm blood, body weight, age and route of administration.

The pharmaceutical preparations of the present invention can be obtained by well known methods such as solutions, mixtures, granules or dragees.

When formulated for oral administration or for mucosal application, the active compounds are formulated with suitable excipients such as carriers, stabilizers or inert diluents and conventionally formulated into suitable dosage forms such as tablets, dragees, capsules, suspensions in water, alcohol, oil or solutions in water, alcohol or oil. Inert carriers which may be used are, for example, Gummi arabicum, magnesium, magnesium carbonate, potassium phosphate, milk sugar, glucose, magnesium stearyl fumarate or starch, in particular maize starch. Preparation can be done by either dry or wet granulation. Oil or carrier oils or solvents such as sunflower oil or fish oil, for example, are acceptable.

Formulations for topical administration may be in the form of a solution in water or in oil, a lotion, an emulsion or a gel, a ointment or a grease, or where appropriate an aerosol, and the adherence may be improved by the addition of a polymer.

The form for nasal administration may be prepared by admixing with conventional additives for this purpose such as stabilizers or inert diluents and conventionally formulated for use as suspensions in water, alcohol or oil, or solutions in water, alcohol or oil. Aqueous compositions for nasal administration may also contain, as additives, components that form chelates, such as ethylenediamine-N, N, N ', N'-tetraacetic acid, citric acid, tartaric acid, or salts thereof. The solutions can be injected into a nasal spray or as a viscous nasal drops, as a gel or cream in the nose.

Pharmaceutical formulations for intravenous, subcutaneous, intradermal or topical administration to the skin are prepared by dissolving, suspending or emulsifying the active compounds or their physiologically acceptable salts, preferably with conventional pharmaceutical auxiliaries, for example, as isotonic or pH adjusting agents, or as solvent carriers. emulsifiers or other excipients.

Because of the short half-lives of some of the drugs described, it is important to use injectable inhibitors. V. _ «T y forms. Such dosage forms may be used, for example, in the form of oil-based crystalline suspensions, microcapsules, rods or implants; these latter can be made from tissue-compatible polymers, namely, biodegradable polymers such as, for example, polylactic acid based on polylactic acid or human albumin.

Concentrations of solutions for topical and inhalation administration are in the range of 0.01 to 5 mg / ml, and for systemic formulations 0.01 to 10 mg / kg.

List of abbreviations

The amino acid abbreviations used correspond to the three letter code accepted by the peptide chemistry described in Eur. J. Biochem., 138, 9 (1984). Other abbreviations used are given below:

ACm - - acetamidomethyl

-Ahx - £ -aminohexanoyl

Aoc - - cis, endo-2-azabicyclo (3.3.0) octane-3-S-carbonyl Boc - - tert-butyloxycarbonyl Being - - tert, -butyl CDF - - chloro- (D) -phenylalanyl Cha - - cyclohexylalanil Chg - - cyclohexylglycyl Cl-Z - 4-chlorobenzyloxycarboxyl DMF - - dimethylformamide DOMT - 0-methyl- (D) -treonyl Dnp - - 2,4-Dinitrophenyl Fmoc - 9-fluorenylmethyloxycarbonyl MbV - O-methyl- (D) -tyrosyl Me - methyl

4-Mebzl - 4-methylbenzyl

Mtw - 4-methoxy-2,3,6-trimethylphenylsulfonyl Mts - mesitylene-2-sulfonyl Nal - 2-naphthylalanil NMP - N-methylpyrrolidine Npg - neopentylglycyl Oic - cis-endo-octahydroindole-2-carbonyl i Opr - isoxazolidin-3-ylcarbonyl Pal - pyridylalanil

Pmc - 2,2,5,7,3-pentamethylchroman-6-sulfonyl

Tbg - tert.-butylglycyl

TFA - trifluoroacetic acid

Thlg - 2-thienylalanil

Those are 4-methylphenylsulfonyl

Tie ~ - 1,2,3,4-Tetrahydroisoquinolin-3-ylcarbonyl

Trt - trityl

The following examples illustrate the most suitable methods for solid phase synthesis of the peptides of the invention; however, the invention is not limited thereto.

The following amino acid derivatives are used:

Fmoc-Arg (Mtr) -OH, Boc- (D) -Arg-OH, Fmoc-Arg (Pmc) -OH, Fmoc-Hyp-OH, Fmoc-Pro-OObt, Fmoc-Gly-OObt, Fmoc-Phe- OObt,

Fmoc-Ser (tBu) -OObt, Fmoc-(D) -Tic-OH, Fmoc-Gln-OH, Fmoc-Aoc-OH,

Fmoc-Thia-OH, Fmoc-Opr-OH, Fmoc- (D) -Asn-OH, Fmoc-B-Ala-OH,

Fmoc-Oic-OH.

example

H- (D) -Arg-Arg-Hyp-Pro-Gly-Leu-Ser- (D) -Tic-Oic-Arg-OH

The peptide is synthesized stepwise using an Applied Biosystems Model 430A peptide synthesizer using the Fmoc method of Novabiocnem based p-benzyloxybenzyl alcohol esterified with Fmoc-Arg (Mtr) -OH (loading about 0.5 mmol / g resin). 1 g of resin is used and synthesis is carried out according to a modified Fmoc-method synthesis program.

The synthesizer cartridges are weighed and each time a 1 mmol amino acid derivative with a free carboxyl group is added together with 0.95 mmol HOObt. Pre-activation is performed directly on the cartridge by dissolving 4 mL of DMF and adding 2 mL of a 0.55 mol / L diisopropylcarbodiimide solution in DMF. The HOObt21 esters of the other amino acids are dissolved in 6 ml of NPM and then all the pre-activated amino acids in situ are combined with a 20% piperidine solution in DMF-e-blocked resin. Upon completion of the synthesis, the use of trifluoroacetic acid and thioanisole and ethanediol as cationic binders results in the cleavage of the peptide from the resin and the removal of the protecting groups from the side chains. After removal of trifluoroacetic acid, the residue is repeatedly rubbed with acetic acid ester and

- (r) Centrifugation. The remainder of the residue is chromatographed over Sephadex; 10% acetic acid is used as eluent. Fractions containing pure peptide are purified and freeze-dried. The desired product is obtained in 43% yield.

Further examples are performed according to the procedure described in Example 1

H- (D) -Arg-Arg-Hyp-Pro-Gly-Cha-Ser- (D) -Tic-Oic-Arg-OH

Product yield 38%.

example

H- (D) -Arg-Arg-Hyp-Pro-Gly-Tbg-Ser- (D) -Tic-Oic-Arg-OH

Product yield 35%.

Claims (5)

1. Novel peptides of general formula I: A-B-C-E-F-K- (D) -Tic - G - M - F '- I (I) A ap) is hydrogen, (Cg-Cg) -alkyl, (C ^ -Οθ) alkanoyl, (C.-C) - alkoxycarbonyl or (C.-C _o) alkylsulfonyl, Wherein 1, 2 or 3 hydrogen atoms, respectively, are optionally substituted with one, two or three identical or different residues, such as carboxyl, amino, (O -, - C,) alkyl, (C, -). C 1) alkylamino, hydroxyl, (C 1 -C 4) acoxy. * -t ± 4, halogen, di (Cg-C ^) - alkylamino, carbamoyl, sulfamoyl, (C.-C.) - alkoxycarbonyl, (C, -C, _) - and aryl (C _r -C N) - aryl- (C ₈ -C ₈ ) -alkyl in which one hydrogen atom is optionally substituted by one residue, such as (C ₈ -C ₆ ) -cycloalkyl, ( _{C 1} -C ₄ ) -alkylsulfonyl, (C ₈ -C ₈ ) -alkyl. ) alkylsulfinyl, (C-C _r. _) -Ar II - (C.-C.) - alkylsulphonylamino onyl, (C, -C, _) - aryl- (C, -C.) -a Ιkilsulfinilas ( Cg-Cg _O ) -aryloxy, (Cg-Cg) -heteroaryl and (Cg-Cg) -heteroaryloxy, and one or two hydrogen atoms being replaced by one or two same or different residues such as carboxyl, amino, (C 1 -C 8). (1-4C) alkylamino, hydroxy, (C 8 -C 6) -alkoxy, halogen, di- (C 8 -C 4) -alkylamino, carbamoyl, sulfamoyl, (C 8 -C 6) -alkoxycarbonyl, (C 8 -C 8) -aryl and ( ^c g ^-C ₁ ^- ₂ ^ -aryl- ( ^{C 8 -C} ₈ ) -alkyl; ^a 2) is a (C₁-C₈) -cycloalkyl, carbamoyl, which, if necessary, the nitrogen atom may be a (Cg-Cg) -alkyl, or ^(c g ± ₂ ^-C ^ aryl substituents, (Cg-cha ^ ₂ —Aryl, (C 1 -C 8) -aryl, (C 8 -C 8) -aryl 23 sulfonyl, (C 8 -C 6) -heteroaryl, or (C 8 -C 6) -heteroarylcyl, wherein a ^) and a ^) in the residues such as aryl, heteroaryl, aroyl, arylsulfonyl and heteroaryl may be optionally substituted with 1, 2, 3 or 4 substituents such as carboxyl, amino, nitro, (Cg-C4) alkylamino, hydroxyl, (Cg) -C 1-4 alkyl, (C 8 -C 7) alkoxy, halogen, cyano, di- (C 8 -C 6) alkylamino, carbamoyl, sulfamoyl and (C 8 -C 4) alkoxycarbonyl, or ag) is a residue. of formula II R ¹ - N - CH - C II 0 (II) where r! is as A under (a) to (g) and (ag); R is hydrogen or methyl; R is hydrogen or (C 8 -C 8) -alkyl, preferably (C 8 -C 6) -alkyl, optionally substituted with one substituent such as amino, substituted amino, hydroxy, carboxyl, carbamoyl, guanidine, substituted guanidine group, ureido group, mercaptogroup, methylmercaptogroup, phenyl, 4-chlorophenyl, 4-fluorophenyl, 4-nitrophenyl, 4-methoxyphenyl, 4-hydroxyphenyl, phthalimido group, 4-imidazolyl, 3-indolyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl or cyclohexyl, wherein the substituted amino group is -NH-A and the substituted guanidine group is -NH-C (NH) -NH-A, wherein A is the same as in (g) or (g); B is a basic amino acid of the L- or D-configuration which may be substituted in the side chain; C are groupings of formulas llla and lllb: - NH - (CH -) - CO 2 n (Illb) is a group of Formula 3872 B where G 'in IV: G '- G' - Gly G (IIla) independently of one another - N - CH C II (IV), where 45 .... ... R and R together with the atoms to which they are attached form a heterocyclic mono-, bi- or tricyclic system of 2 to 15 atoms and n = 2 to 8; E is a neutral, acidic or basic, aliphatic or alicyclic-aliphatic amino acid residue; F, independently of one another, is a neutral, acidic or basic, aliphatic or aromatic amino acid residue which may be substituted in the side chain or is a direct bond; (D) -Tic is a residue of formula V: G is the same as G 'having the meanings given above or is a direct bond; F 'is the same as F is a -NH (CH ₂ ) _n - residue where n = 2-3, or, if G is not a direct bond, may be a linear bond; I is -OH, -NH ₂ or -NHC ₂ H ₅ ; K is a residue of -NH- (CH-) -CO-, where x = 1-4, or is a direct X bond, and M has the same meanings as F and the physiologically acceptable salts of these compounds. The peptide of formula I according to claim 1. characterized in that the resulting peptide B is Arg, Lys, Orn, 2,4-diaminobutyryl or a residue of L-homoarginine, wherein in each case the guanidine or amino groups of the side chain may be replaced by a group as defined in (a) and (a) 2 (l) of the invention; E is an aliphatic or alicyclic aliphatic amino acid residue of the L- or D-configuration which has 1-14 C-atoms in the side chain; F is a residue of a basic amino acid of the L- or D-configuration, such as Arg or Lys, wherein the guanidine group or amino group of the side chain may be replaced by A as described in claim 1 of the invention. a ^) or a ^) moieties, or is a -NH-CCHg) ^ residue, where n = 2-8; K is a residue of -NH- (CHg) -CO-, where x = 2-4, or a direct bond. The peptide of formula I according to claims 1 and / or 2. characterized in that the resulting peptide B is Arg, Orn or Lys, wherein the guanidine or amino groups on the side chain are unsubstituted or may be substituted such as (C 1 -C 8) -alkanoyl, (C 1 -C 6) -aryl, heteroaryl, (C (C 1 -C 8) -alkylsulfonyl or (C 8 -C 8) -arylsulfonyl wherein the aryl, heteroaryl, aroyl, arylsulfonyl and heteroaryl moieties as described in (g) may optionally be 1, 2, 3 or 4; or different substituents; E is leucine, isoleucine, norleucine, tert-butylglycine, serine, threonine, or cyclohexylalanine; K is a direct connection and M is a direct connection. 4. A peptide of the formula I according to claims 1-3, characterized in that in the resulting peptide A is hydrogen, (D) - or (L) -H-Arg, (D) - or (L) -H-Lys or (D) - or (L) -H-Orn; B is Arg, Orn or Lys, where the guanidino side chain, or the amine may be substituted with a hydrogen, ^(C 1 ~ ^C 8) alkanoyl, (C ^ -C ^ g) -aroilas (Cg-C _g) - heteroaryl, (C 1 -C ₈ ) -alkylsulfonyl or (C ₈ -C ₁₂ ) -arylsulfonyl whose aryl, heteroaryl, aroyl, arylsulfonyl and heteroaryl moieties may be optionally substituted with 1, 2, 3 or 4 substituents such as as methyl, methoxy and halogen; C is Pro-Pro-Gly, Hyp-Pro-Gly or Pro-Hyp-Gly; E is Leu, Ile, Tbg, or Cha; F is Ser, hSer, Lys, Leu, Vai, Nle, Ile or Thr; K and M are direct bonds; G is a residue of the heterocyclic system of formula IV, wherein the most suitable residues are pyrrolidine-2-carbonic acid, piperidine-2-carbonic acid, tetrahydroisoquinoline-3-carbonic acid, eis- and trans-decahydroisoquinoline-3-carbonic acid, cis- endo, cis-exo, trans-octahydrocyclopenta (b) pyrrole-2-carbonic acid or hydroxyproline-2-carbonic acid; F ' 5. is Arg and I is OH. for utilizes Formulas I peptide according to 1-4 points, ti how drug substance. 6th Formulas I peptide according to 1-4 points, for utilizes ie pathological conditions caused by bradykinin and a bradykinin-related peptide. ' 7. Pharmaceutical compositions comprising, as appropriate, a pharmaceutically acceptable carrier, wherein the active ingredient is a compound of the formula I according to claims 1-4.