DE10200666A1 - New peptide compounds containing upto 6 aminoacid residues, are potent and specific thrombin inhibitors used for treating and diagnosing diseases associated with thrombin - Google Patents

Abstract

Peptide compounds (I), containing upto 6 aminoacid residues, are new. Peptide compounds of formula Y 1>-(NH-X 1>-CO)-(NH-X 2>-CO)-(NH-X 3>-CO)-(NH-X 4>-CO)-(NH-X 5>-CO)-(NH-X 6>-CO)-Y 2>(I), their derivatives truncated by at least one aminoacid at the C- and/or N-terminal and salts, are new. Y 1H, Me, acetyl or a group having a 1-32C chain as backbone; NH-X 1-CO : basic aminoacid residue, preferably Arg, Lys or Orn; NH-X 2-CO : cyclic, non-polar aminoacid residue, preferably cyclohexylalanine (Cha) or cyclohexylglycine; NH-X 3-CO : any aminoacid residue, preferably Nle, Leu, Ile, Cha, cyclohexylglycine, Pro, Asp or Glu; NH-X 4-CO : cyclic aminoacid residue, preferably Cha, cyclohexylglycine, Tyr or Phe; NH-X 5-CO : aminoacid residue with a polar side-chain, preferably Gln, Orn, Glu, Arg, Lys, Asn or Asp, or is replaced by a direct bond; NH-X 6-CO : any aminoacid residue, preferably Arg, or is replaced by a direct bond; Y 2OH (C-terminal carboxy group), NH 2(C-terminal amide group), H (C-terminal aldehyde group), 7-amido-4-methyl-coumarin (bonded via the carboxy group), p-nitroanilide (bonded via the carboxy group) or the residue of a compound with a 1-35 atom chain; all aminoacid residues are in L- or D-isomer form. In alternative definitions of the substituents of (I), Y 1and Y 2are as above and: (a) NH-X 1-CO : any aminoacid residue, preferably Val, Ala, Leu, Ile, Nle, Asp, Glu, Ser, Thr, Tyr, Arg, Lys or Orn, or is replaced by a direct bond; NH-X 2-CO : any aminoacid residue, preferably Ala, Val, Leu, Ile, Nle, Ser, Thr, Tyr, Pro, citrulline, Arg, Lys, Orn, Cha or cyclohexylglycine, or is replaced by a direct bond; NH-X 3-CO : any aminoacid residue, preferably Cha or cyclohexylglycine; NH-X 4-CO : small aminoacid residue, preferably Pro, or is replaced by a direct bond; NH-X 5-CO : any aminoacid residue, preferably Tyr or Phe, or is replaced by a direct bond; NH-X 6-CO : basic aminoacid residue, preferably Arg, Lys or Orn; (b) NH-X 1-CO : as in (a); NH-X 2-CO : any aminoacid residue, preferably Ala, Val, Leu, Ile, Nle, Ser, Thr, Tyr, Pro, citrulline, Arg, Lys, Orn, His, Glu, Asp, Trp, Cha or cyclohexylglycine, or is replaced by a direct bond; X 3-X 6as in (a); (c) X 1-X 3as in (b); NH-X 4-CO : small aminoacid residue, preferably Pro or azetidine-2-carboxylic acid; NH-X 5-CO : aromatic aminoacid residue, preferably Tyr or Phe; NH-X 6-CO : basic aminoacid residue, preferably Arg, Lys, Orn or homoarginine; (d) NH-X 1-CO : any aminoacid residue, preferably Val, Ala, Leu, Ile, Nle, Asn, Gln, Ser, Thr, Tyr, Arg, Lys, Orn, Phe, dichlorophenylalanine, tetrahydronorharman-3-carboxylic acid, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, 4-phenyl-piperidine-4-carboxylic acid, thienylalanine, phenylglycine or p-nitirophenylalanine, or is replaced by a direct bond; X 2as in (b); X 3as in (a), and X 4-X 6as in (c); all aminoacid residues are in L- or D-isomer form. ACTIVITY : Anticoagulant; Thrombolytic; Vasotropic; Antiarteriosclerotic; Antiinflammatory; Cytostatic; Neuroprotective. MECHANISM OF ACTION : Thrombin inhibitor. N-Acetyl-L-Phg-D-Cha-L-Aze-D-Tyr-L-Har-amide (Ia) inhibited thrombin by 95% at a concentration of 250 nM.

Description

This invention relates to biological agents that interact with and inhibit thrombin. The active ingredients are useful as anticoagulants for humans and animals. In particular, this invention extends to small peptidic molecules of length 3-6 amino acids of the general formula (I) Y ¹ - (NH-X ¹ -C =O) - (NH-X ² -C =O) - (NH-X ³ -C =O) - (NH-X ⁴ -C = O) - (NH ³ ) X ⁵ -C = O) - (NH-X ⁶ -C = O) -Y ² (I) in which Y ¹ , Y ² and X ^{1-6 have} the meanings given in the description, or N- or C-terminally shortened variants of formula (I) having a high antithrombotic activity. The invention also relates to compositions and combinations with these substances for therapeutic, prophylactic and diagnostic purposes.

acute vascular diseases, myocordial infarction, stroke, pulmonary embolism, deep vein thrombosis or peripheral vascular obstruction and other thrombosis of the blood system, constitute a significant health hazard. Such disorders are caused by complete or partial constipation of the blood vessel a plug containing fibrin and platelets.

Current methods The treatment and prophylaxis of such thrombosis diseases include therapeutics, which work in two different ways. The first type of therapeutics prevents thrombin activity or thrombosis formation and thus the formation of the plug. These Drugs also prevent the development of platelets and their aggregation. The second category of drugs is accelerating the thrombolysis and dissolves The plug on, thus eliminating this from the blood vessel and lifts the Blockage of blood flow up.

heparin and coumarin, preparations of the first type will be in great Measurements to Treatment of venous Thrombosis embolism used in the thrombotic activity for development and expansion of the thrombus is responsible. Although effective, However, heparin gets many unwanted ones Side effects such as bleeding or thrombocytopenia. The same applies to Coumarin by blocking or preventing the formation of Prothrombin works and for some time until its full effect needed. Taken together, this has led to a search for more specific ones and less toxic anticoagulants, e.g. peptidic Inhibitors.

Hirudin is a naturally occurring polypeptide produced by the leech Hirudo medicinalis. This active ingredient, which is synthesized in the salivary gland of the leech, is the most effective known natural anticoagulant. Hirudin is a direct thrombin inhibitor and inhibits coagulation of the blood by strong binding to thrombin (Kd = 2 × 10 ^-14 M) in a stoichiometric 1: 1 complex [Stone & Hofstenge, Kinetics of the inhibition of thrombin by hirudin, Biochemistry 25, pp. 4622-4628 (1986)]. This, in turn, prevents the thrombin from catalyzing the conversion of fibrinogen to fibrin (grafting), as well as preventing any other thrombin-mediated cleavage processes.

Animal studies have demonstrated the efficacy of hirudin, purified from leeches, in the prevention of venous thrombosis, vascular congestion, and thrombin-induced widespread intravascular coagulation. In addition, hirudin exhibits low toxicity, low antibody production and rapid clearance from the bloodstream [F.Markwardt et al., Pharmacological studies on the antithrombic action of hirudin in experimental animals, Thromb. Haemost. 47, pp. 226-229 (1982)]. In projects aimed at producing larger amounts of hirudin, attempts have been made to produce the polypeptide by recombinant DNA technology. The presence of an O-sulfated tyrosine residue in natural hirudin and the inability of the microorganisms to make a similar modification made the prospect of recombinant production of biologically active hirudin highly speculative. The observation that desulfated hirudin is almost as effective as its sulfated counterpart [US Pat. 4,654,302], has pioneered cloning and expression in various expression systems, including S. cerevisiae [Harvey et al., Cloning and expression of cDNA coding for the anticoagulant hirudin from the bloodsucking leech, Hirudo medicinalis, PNAS 83, p 1084-1088; Europ.Pat.Appl. 158,654, 168,342 and 171024], E. coli [Bergmann et al., Chemical synthesis and expression of a coding for hirudin, the thrombin-specific inhibitor from the leech Hirudo medicinalis, Biol. Chem. Hoppe-Seyler 367, p 731-740; Europ.Pat.Appl. 200 655] and on the tips of a filamentous phage as a fusion protein with protein III (pIII) [Wirsching et al., Display of functional thrombin inhibitor hirudin on the surface of phage M13, Gene 204, pp. 177-184]. Despite these advances, hirudin is still quite expensive to produce. Still has It has undergone the third clinical phase and has recently been approved for the treatment of heparin-induced thrombocytopenia (HMR).

More recently, the identification of peptide fragments of natural hirudin has been successful, as well as prolonging the coagulation time. However, such peptide fragments can not be fully satisfactory because of their low effectiveness in preventing grafting. For example, N-acetyl-hirudin _45-65 has four orders of magnitude lower activity than natural hirudin, although it is still a relatively large molecule. The problem of rather low affinities for thrombin has been explained by the development of Hirulogen [US Pat. No. 5 433 940]. These molecules mimic the action of hirudin by binding to both the outward anion-binding side of low affinity and to the catalytic side of α-thrombin. Thus, hiruloges are characterized by a half associated with the anion-binding thrombin exterior, a linking group, and a portion directed against the catalytic site of thrombin. The most preferred Hirulog is Hirulog-8, a 20 amino acid peptide consisting of the catalytic center inhibiting peptide D-Phe-Pro-Arg-Pro, a Gly ₄ junction sequence and the sequence -Asn-Gly- Asp-Phe-Glu-Glu-Ile-Pro-Glu-Glu-Tyr-Leu-OH of hirudin is constructed. Hirulog-8 has recently entered the US market.

Despite the progress of relatively high efficacy for thrombin (K _i = 2.3 nM), hirulogues are still relatively large molecules, resulting in relatively cumbersome schemes. as mixed heterologous / solid phases, must be synthesized. Like hirudin, hirologists are only parenterally helpful and must be monitored carefully. Therefore, Hiruloge are not suitable as lead structures for small molecules, which could eventually be administered orally.

Therefore, there has been some effort to identify smaller peptides as potent inhibitors of thrombin. Already in 1956 Bettelheim showed that the fibrinopeptide A comparably inhibits the reaction between thrombin and fibrinogen. In a joint research by Blombäck and Nobel Pharma / Kabi in Stockholm, fibrinopeptide A-derived peptide sequences having not more than nine amino acids have been found to be effective on thrombin. Essential components of the efficacy were an N-terminal Phe and a C-terminal Arg separated by seven amino acids. Fewer amino acids reduced potency, but surprisingly, a tripeptide with N- and C-terminal Phe and Arg, respectively, showed excellent potency. The best tripeptide inhibiting the thrombin-fibrinogen reaction was Bz-Phe-Val-Arg-OMe, with Val preceding the Arg as in the full-length fibrinopeptide [Blombäck et al., Synthetic peptides with anticoagulant and vodorizing activity, Scand. J. Clin. Lab. Invest. 24, pp. 59-66 (1969), US Pat. No. 3,826,794 (1974)]. In contrast to fibrinopeptide A, Pro precedes Arg in a number of other thrombin clipping regions, such as prothrombin, factor XIII and human growth hormones. Based on the Pro-Arg sequence, most of the most effective thrombin-inhibiting peptides and peptidomimetics were developed. Among these most potent inhibitors is HD-Phe-Pro-Arg-H (K _i = 70 nM) [Bajuz et al., Inhibition of thrombin and trypsin by tripeptide aldehydes, Int. J. Peptide Protein Res. 12, pp. 217-221 (1978); Hung.Pat. 169870 (1974)]. The idea for this peptide aldehyde arose from the discovery of peptide aldehydes of bacterial origin by H. Umezawa. These so-called leupeptides (eg, Ac-Leu-Leu-Arg-H) are inhibitors of plasmin and other trypsin-like proteases, but not thrombin. The aldehyde carbon in its acetal form has a tetrahedral structure, the same as the carbonyl carbon of substrates in the transition phase.

From these aldehydes just mentioned, Shaw et al. the irreversible chloromethyl ketone inhibitor HD-Phe-Pro-Arg-CH ₂ -Cl with a K _i of 25 nM [Kettner et al., HD-Phe-Pro-Arg-CH ₂ -Cl - a selective activity label for thrombin, Thromb. Res. 14, pp. 969-973 (1979)]. Development work at Eli Lilly has resulted in N-methyl-D-Phe-Pro-Arg-H, also known as Efegatran [tm]. The D-Phe-Pro-Arg sequence has recently been further developed. Speculation that an N-terminal amino acid with aromatic / lipophilic groups might give greater efficacy against thrombin has led to the discovery of several new amino acid inhibitors at this position, including β-β-diphenylalanine (Dpa), phenylglycine, cyclohexylglycine, carboxy- 1,2,3,4-tetrahydroisoquinoline (Tiq) [Schuman et al., Highly selective thrombin inhibitors, J. Med. Chem. 36, pp. 314-319 (1993)]. The most interesting compound was D-1-Tiq-Pro-Arg-H, which gave twice the efficacy compared to Boc-D-Phe-Pro-Arg-H. However, trypsin is inhibited to the same extent as thrombin.

Out accessible today Data is clear that, although there are some effective anticoagulant compounds There is a need for powerful Antithrombin agents that act quickly to stop the grafting and which do not interfere with other protease activities, e.g. the plasmin effect on dissolution of the plug interfere.

outgoing of this prior art, the present invention was the The object is to provide compounds that are in the sense of a Thrombin inhibition are biologically active and the disadvantages of the previous avoid prior art described. The underlying of the invention There was also a problem therein, thrombin specifically with low drug concentrations and low cell toxicity to inhibit.

• 1. ein Wasserstoff oder
• 2. eine Methylgruppe oder
• 3. eine Acetylgruppe ist oder
• 4. durch ein Rückgrat aus einer Kette von 1 bis 32 Kohlenstoffatomen charakterisiert ist,

wobei (NH-X¹-C=O) eine D- oder L-Aminosäure, vorzugsweise

• 1. Valin oder
• 2. Alanin oder
• 3. Leucin oder
• 4. Isoleucin oder
• 5. Norleucin oder
• 6. Asparagin oder
• 7. Glutamin oder
• 8. Serin oder
• 9. Threonin oder
• 10. Tyrosin oder
• 11. Arginin oder
• 12. Lysin oder
• 13. Ornithin oder
• 14. Phenylalanin oder
• 15. Dichlorphenylalanin oder
• 16. Tetrahydronorharman-3-carbonsäure oder
• 17. Tetrahydroisoquinolin(1,2,3,4)-3-carbonsäure oder
• 18. 4-Phenylpiperidin-4-carbonsäure oder
• 19. Thienylalanin oder
• 20. Phenylglycin oder
• 21. p-Nitrophenylalanin ist
• 22. oder durch eine chemische Bindung ersetzt wird,

wobei (NH-X²-C=O) eine D- oder L-Aminosäure, vorzugsweise

• 1. Alanin oder
• 2. Valin oder
• 3. Leucin oder
• 4. Isoleucin oder
• 5. Norleucin oder
• 6. Serin oder
• 7. Threonin oder
• 8. Tyrosin oder
• 9. Prolin oder
• 10. Citrullin oder
• 11. Arginin oder
• 12. Lysin oder
• 13. Ornithin oder
• 14. Histidin oder
• 15. Glutaminsäure oder
• 16. Asparaginsäure oder
• 17. Tryptophan oder
• 18. Cyclohexylalanin oder
• 19. Cyclohexylglycin ist
• 20. oder durch eine chemische Bindung ersetzt wird,

wobei (NH-X³-C=O) eine beliebige Aminosäure. beispielsweise

• 1. L-Cyclohexylalanin oder
• 2. D-Cyclohexylalanin oder
• 3. L-Cyclohexylglycin oder
• 4. D-Cyclohexylglycin ist,

wobei (NH-X⁴-C=O) eine kleine Aminosäure, vorzugsweise

• 1. L-Prolin oder
• 2. D-Prolin oder
• 3. L-Azetidin-2-carbonsäure oder
• 4. D-Azetidin-2-carbonsäure ist,

wobei (NH-X⁵-C=O) eine vorzugsweise aromatische Aminosäure, wie

• 1. L-Tyrosin oder
• 2. D-Tyrosin oder
• 3. L-Phenylalanin oder
• 4. D-Phenylalanin ist,

wobei (NH-X⁶-C=O) eine Aminosäure mit basischer Seitenkette, vorzugsweise

• 1. L-Arginin oder
• 2. D-Arginin oder
• 3. L-Lysin oder
• 4. D-Lysin oder
• 5. L-Ornithin oder
• 6. D-Ornithin oder
• 7. L-Homoarginin oder
• 8. D-Homoarginin ist,

wobei Y² entweder

• 1. eine OH-Gruppe (die C-endständige Aminosäure hat eine endständige Carbonsäure-Gruppe) oder
• 2. eine Aminogruppe (bei der C-endständigen Aminosäure ist die Carbonsäure- durch eine Amid-Gruppe ersetzt) oder
• 3. ein Wasserstoff (bei der C-endständigen Aminosäure ist die Carbonsäure- durch eine Aldehyd-Gruppe ersetzt) oder
• 4. 7- Amido-4-methylcumarin (über die Carbonsäuregruppe kombiniert) oder
• 5. Paranitroanilid (über die Carbonsäuregruppe kombiniert) oder
• 6. durch eine Verbindungskette aus eins bis 35 Atomen ersetzt ist,

oder ein am C-Terminus und/oder am N-Terminus um nicht weniger als eine Aminosäure verkürztes Molekül und pharmazeutisch akzeptable Salze davon.According to the invention, this object is achieved by the compound of the formula Y ¹ - (NH-X ¹ -C =O) - (NH-X ² -C =O) - (NH-X ³ -C =O) - (NH-X ⁴ -C = O) - (NH ³ ) X ⁵ -C = O) - (NH-X ⁶ -C = O) -Y ² (I) solved,
where Y ^{1 is} either

• 1. a hydrogen or
• 2. a methyl group or
• 3. is an acetyl group or
• 4. characterized by a backbone of a chain of 1 to 32 carbon atoms,

where (NH-X ¹ -C = O) is a D- or L-amino acid, preferably

• 1. Valine or
• 2. Alanine or
• 3. Leucine or
• 4. Isoleucine or
• 5. norleucine or
• 6. asparagine or
• 7. Glutamine or
• 8. Serine or
• 9. Threonine or
• 10. Tyrosine or
• 11. Arginine or
• 12. Lysine or
• 13. Ornithine or
• 14. Phenylalanine or
• 15. dichlorophenylalanine or
• 16. Tetrahydronorharmane-3-carboxylic acid or
• 17. Tetrahydroisoquinoline (1,2,3,4) -3-carboxylic acid or
• 18. 4-phenylpiperidine-4-carboxylic acid or
• 19. thienylalanine or
• 20. phenylglycine or
• 21. p-nitrophenylalanine
• 22. or is replaced by a chemical bond,

where (NH-X ² -C = O) is a D- or L-amino acid, preferably

• 1. Alanine or
• 2. Valine or
• 3. Leucine or
• 4. Isoleucine or
• 5. norleucine or
• 6. Serine or
• 7. Threonine or
• 8. Tyrosine or
• 9. Proline or
• 10. Citrulline or
• 11. Arginine or
• 12. Lysine or
• 13. Ornithine or
• 14. histidine or
• 15. glutamic acid or
• 16. aspartic acid or
• 17. Tryptophan or
• 18. cyclohexylalanine or
• 19. Cyclohexylglycine is
• 20. or is replaced by a chemical bond,

where (NH-X ³ -C = O) is any amino acid. for example

• 1. L-cyclohexylalanine or
• 2. D-cyclohexylalanine or
• 3. L-cyclohexylglycine or
• 4. D-cyclohexylglycine is,

where (NH-X ⁴ -C = O) is a small amino acid, preferably

• 1. L-proline or
• 2. D-proline or
• 3. L-azetidine-2-carboxylic acid or
• 4. D-azetidine-2-carboxylic acid is,

where (NH-X ⁵ -C = O) is a preferably aromatic amino acid, such as

• 1. L-Tyrosine or
• 2. D-tyrosine or
• 3. L-phenylalanine or
• 4. D-phenylalanine is,

where (NH-X ⁶ -C = O) an amino acid with a basic side chain, preferably

• 1. L-arginine or
• 2. D-arginine or
• 3. L-lysine or
• 4. D-lysine or
• 5. L-ornithine or
• 6. D-ornithine or
• 7. L-homoarginine or
• 8. D-homoarginine is,

where Y ^{2 is} either

• 1. an OH group (the C-terminal amino acid has a terminal carboxylic acid group) or
• 2. an amino group (in the C-terminal amino acid, the carboxylic acid is replaced by an amide group) or
• 3. a hydrogen (in the C-terminal amino acid, the carboxylic acid is replaced by an aldehyde group) or
• 4. 7- Amido-4-methylcoumarin (combined via the carboxylic acid group) or
• 5. Paranitroanilide (combined via the carboxylic acid group) or
• 6. is replaced by a linking chain of one to 35 atoms,

or a molecule shortened at the C-terminus and / or at the N-terminus by not less than one amino acid and pharmaceutically acceptable salts thereof.

The The invention also relates to derivatives of the above-mentioned compounds.

Particularly advantageous results are achieved when the peptide according to the invention is N-acetyl-R ₁ -L-Cha-D-Pro-D-Tyr-L-Arg-amide, where R _{1 is} D-Gln-D- His, D-Glu, D-Val-D-His, L-Ala, L-Ile-L-Arg, L-Tyr-L-Cit, L-Ser-L-Ser, D-Val, L-Trp, L-Ser-L-Ala, L-Ser-L-Arg, D-Lys-L-Ile, D-Tyr, L-Arg or L-Tyr-D-Pro, when the peptide according to the invention is N Acetyl-L-Ala-D-Cha-L-Aze-D-Tyr-L-Arg-amide to give N-acetyl-L-Ala-D-Cha-L-Pro-D-Tyr-L-Har. amide or N-acetyl-R ₂ -D-Cha-L-Aze-D-Tyr-L-Har-amide, where R _{2 is} L-Trp, L-Ala, D-Phe, L-Dcp, L -Nhm, L-Iq3, L-Ppd, L-Tea, L-Phg, L-Nle, L-Cha or L-Pnp.

The compounds of the invention can be used to inhibit all thrombin-mediated or thrombin-associated functions and processes. Pharmaceutical compositions containing these compounds, as well as methods for the treatment and prophylaxis of vascular diseases, inflammatory reactions, carcinomas and neurodegenerative diseases using these compounds are also provided by the invention. The compounds can also be used for ex vivo imaging, for storage and treatment of blood outside the body and for coating in vivo vasive devices. Further, the compounds may be administered to a patient (a patient is herein to be understood as a human or animal) in combination with a fibrinolytic agent to increase the efficacy of a given dose or reduce the dose necessary to achieve a desired effect, such as dissolving and Blood plug or the prevention of re-blockage of the previously blocked blood vessel.

by virtue of their high potential and the fact that they use chemical synthesis techniques can be produced can the connections reasonably priced be produced in commercially practicable quantities. The peptides are converted into suitable salt forms such as acetates and sulfates.

Furthermore the molecules of the invention are essential smaller than hirudin and the other peptides previously described Thrombin inhibitor. Therefore, it is unlikely an undesirable reaction of the immune system in patients treated with these substances cause. Accordingly, the use of these thrombin inhibitors not limited to the treatment of acute illnesses. These Connections can also in the treatment of chronic thromboembolic diseases such as arteriosclerosis or restenosis due to angioplasty, be used. The compounds of the invention may also in a variety of other applications instead of natural and recombinant thrombin.

As one can conclude from the disclosure, are the compounds of the invention, Compilations and procedures useful for treatment and prevention various diseases related to unwanted Effects of thrombin, as well as for diagnostic purposes.

Finally, should mentioned be that the molecules of this invention as lead structures for the development of molecules with even more advantageous properties in view of the above Applications can serve.

pharmaceutical Acceptable salts of peptides of this invention include those disclosed in U.S. Pat Acid addition generated Salts made from inorganic acids and carboxylic acids be formed. The compounds represented by the formula (I) are prepared by known methods of peptide coupling.

In a preferred embodiment are the compounds of the invention as a compound mixture, which by its content of at least two of the compounds of the invention is characterized. Preferred pharmaceutically acceptable salts The compounds are formed with an inorganic acid. Particularly preferred is the formation of a pharmaceutically acceptable Salt with hydrochloric acid, Chlorine acid, hydrobromic, bromic and / or another halogen acid. Another particularly preferred embodiment is the formation a pharmaceutically acceptable salt with sulfuric acid and / or Phosphoric acid. Advantageously, a pharmaceutically acceptable salt with a organic acid be formed. Particularly preferred is the formation of the pharmaceutical acceptable salt with acetic acid, propionic acid, malonic, maleic acid, Citric acid, succinic acid, fumaric acid, malic acid, benzoic acid, and / or a similar one Carboxylic acids. The by acid addition formed salts are prepared in a conventional manner, e.g. by neutralizing the free base form of the compound (I) with the acid.

The Compounds according to the invention can in compounds and methods of inhibiting all thrombin-mediated or thrombin-associated functions. pharmaceutical Compositions containing these molecules are, as well Methods of treatment and prophylaxis of vascular, inflammatory Reactions, carcinomas and neurodegenerative diseases that use these compounds, also part of the invention. The Substances can also be used in composition for ex vivo imaging, for storage and treatment of blood outside the body and for coating invasive devices. Furthermore, the Compounds of the invention a patient (a patient becomes a human or an animal understood) in combination with a fibrinolytic agent to increase the effectiveness of a given dose or to achieve a desired Effect, like dissolving a blood plug or preventing the clogging of the previously blocked blood vessel necessary Reduce the dose.

by virtue of their high potential and the fact that they use chemical synthesis techniques can be produced are the substances according to the invention inexpensive producible in a commercially practicable amount. To be favoured the peptides are converted into suitable salt forms, such as acetates or sulfates.

The The invention also relates to a medicament characterized by its content on one or more compounds of the invention with the usual Carriers, Auxiliary agents or additives is characterized. Further is also a diagnostic composition containing one or several of the compounds of the invention.

One Another object of the invention is the use of the Compound as a thrombin inhibitor and for the preparation of a medicament for thrombin inhibition, inhibition of fibrin formation and / or for Inhibition of the formation of a coagulation plug.

The use of one or more of the compounds for the preparation of a diagnostic composition is also provided by the invention. In one of the particularly preferred embodiments, a compound according to the invention is used for the preparation of the diagnostic composition in which Y ^{2 is} 7-amido-3-methylcoumarin or paranitroanilide in the formula (I).

The Compounds of the invention opposite previously known thrombin inhibitors diverse Advantages. In particular, the peptides are easily synthesized, already slightly modified and have a high activity with high specificity at the same time and low toxicity on. About that In addition, the small peptides, unlike hirudin and hirolog, serve as lead structures for preferably orally available active ingredients.

The The following examples illustrate the invention without limiting it.

example 1

N-acetyl-D-Gln-D-His-L-Cha-D-Pro-D-Tyr-L-Arg-amide

This peptide was prepared by solid-phase synthesis using an ABIMED AMS 96 synthesizer (ABIMED Analysen-Technik GmbH, Langenfeld, Germany). In detail, 1 meq Rink amide resin was sequentially reacted with 2 x 5 meq of protected amino acid. The activation was carried out with 2 × 5 meq of TBTU (O-benzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium tetrafluoroborate. After up to 6 cycles of synthesis, the N-terminus was acetylated with acetic anhydride. Then the protection of the peptide was abolished by treatment with 90% TFA, 2.5% triisopropylsilane, 2.5% H ₂ O and 5% dichloromethane. In the same step, the decoupling of the peptide was carried out by its carrier. The test compound was then dissolved in 20 μl of trifluoroacetic acid in a drying step and then incubated with 2 × 750 μl cold butyl ether at -20 ° C. After centrifugation, the supernatant was removed and the remainder ether evaporated. The identity of the products was randomly confirmed by mass spectroscopy.

Inhibition of thrombin was determined by in vitro inhibition of thrombin amidase activity and is 59% at a peptide concentration of 1 μM. The inhibitory constant K _i values were obtained from assays in which thrombin hydrolyzes the fluorogenic substrate Tos-Gly-Pro-Arg- (7-amino-4-methylcoumarin). The assays were performed in 30 μl of assay buffer (0.05 M Tris, 0.1 M NaCl, 0.1% PEG 8000, pH 7.6) with 10 μl of human thrombin solution (10 ^-5 U / μl in assay buffer) and 140 μl a solution of the fluorogenic substrate in an assay buffer at a concentration of 30 μM. Solutions of the test compound (10 μl) were added at various concentrations. The rates of hydrolysis of the substrate were measured by monitoring the reactions at 460 nm of the release of 7-amino-4-methylcoumarin using AMC. The reaction reached equilibrium within three minutes after thrombin was mixed with the substrate and an inhibitor. The kinetic data of the competitive inhibition (K _m , V _max and K _i ) were analyzed by the Hanes plot (A / V versus A at different values of A, where A is the substrate concentration and V is the reaction rate).

Example 2

N-acetyl-D-Glu-L-Cha-D-Pro-D-Tyr-L-Arg-amide

This Peptide was synthesized as described in Example 1 and for use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 1 μM 58%.

Example 3

N-Acetyl-D-Val-D-His-L-Cha-D-Pro-D-Tyr-L-Arg-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 1 μM 51%.

Example 4

N-acetyl-L-Ala-L-Cha-D-Pro-D-Tyr-L-Arg-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 1 μM 44%.

Example 5

N-acetyl-L-Ile-L-Arg-L-Cha-D-Pro-D-Tyr-L-Arg-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 1 μM 61%.

Example 6

N-Acetyl-L-Tyr-L-Cit-L-Cha-D-Pro-D-Tyr-L-Arg-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 1 μM 55%.

Example 7

N-acetyl-L-Ser-Ser-L-L-Cha-D-Pro-D-Tyr-L-Arg amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 1 μM 40%.

Example 8

N-Acetyl-D-Val-L-Cha-D-Pro-D-Tyr-L-Arg-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 1 μM 45%.

Example 9

N-acetyl-L-Trp-L-Cha-D-Pro-D-Tyr-L-Arg-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 1 μM 63%.

Example 10

N-acetyl-L-Ser-L-Ala-L-Cha-D-Pro-D-Tvr-L-Arg amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 1 μM 47%.

Example 11

N-acetyl-L-Ser-L-Arg-L-Cha-D-Pro-D-Tyr-L-Arg amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 1 μM 51%.

Example 12

N-acetyl-D-Lys-L-Nle-L-Cha-D-Pro-D-Tyr-L-Arg-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 1 μM 48%.

Example 13

N-acetyl-D-Tyr-L-Cha-D-Pro-D-Tyr-L-Arg-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 1 μM 47%.

Example 14

N-acetyl-L-Arg-L-Cha-D-Pro-D-Tyr-L-Arg-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 1 μM 47%.

Example 15

N-Acetyl-L-Tyr-D-Pro-L-Cha-D-Pro-D-Tyr-L-Arg-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 1 μM 44%.

Example 16

N-acetyl-L-Ala-D-Cha-L-Aze-D-Tyr-L-Arg-amide

sDieses Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 1 μM 63%.

Example 17

N-acetyl-L-Ala-D-Cha-L-Pro-D-Tyr-L-Har-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 1 μM 56%.

Example 18

N-acetyl-L-Trp-D-Cha-L-Aze-D-Tyr-L-Har-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 250 μM 76%.

Example 19

N-acetyl-L-Ala-D-Cha-L-Aze-D-Tyr-L-Har-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 250 μM 77%.

Example 20

N-acetyl-D-Phe-D-Cha-L-Aze-D-Tyr-L-Har-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 250 μM 77%.

Example 21

N-acetyl-L-Dcp-D-Cha-L-Aze-D-Tyr-L-Har-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 250 nM 75%.

Example 22

N-acetyl-L-NHM-D-Cha-L-Aze-D-Tyr-L-Har-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 250 nM 80%.

Example 23

N-acetyl-L-Iq3-D-Cha-L-Aze-D-Tyr-L-Har-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 250 nM 72%.

Example 24

N-acetyl-L-Ppd-D-Cha-L-Aze-D-Tyr-L-Har-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 250 nM 76%.

Example 25

N-acetyl-L-Tea-D-Cha-L-Aze-D-Tyr-L-Har-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 250 nM 74%.

Example 26

N-acetyl-L-Phg-D-Cha-L-Aze-D-Tyr-L-Har-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 250 nM 95%.

Example 27

N-acetyl-L-Nle-D-Cha-L-Aze-D-Tyr-L-Har-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 250 nM 89%.

Example 28

N-acetyl-L-Cha-D-Cha-L-Aze-D-Tyr-L-Har-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 250 nM 90%.

Example 29

N-acetyl-L-Pnp-D-Cha-L-Aze-D-Tyr-L-Har-amide

This Peptide was synthesized as described in Example 1 and for the Use prepared in the assay.

The Inhibition of thrombin was confirmed by in vitro inhibition of thrombin amidase activity in exemplary embodiment 1 determines and amounts at a peptide concentration of 250 nM 72%.

description the abbreviations

Ala

= Alanine

Val

= Valine

Leu

= Leucine

Ile

= Isoleucine

Per

= Proline

Phe

= Phenylalanine

phg

= Phenylglycine

Cha

= Cyclohexylalanine

Trp

= Tryptophan

mead

= Methionine

Gly

= Glycine

Ser

= Serine

Thr

= Threonine

Cys

= Cysteine

Tyr

= Tyrosine

Asn

= Asparagine

Gln

= Glutamine

Asp

= Aspartic acid

Glu

= Glutamic acid

Lys

= Lysine

bad

= Arginine

His

= Histidine

Nle

= Norleucine

Orn

= Ornithine

Cit

= Citrulline

Aze

= Azetidine

Har

= Homoarginine

dcp

= Dichlorophenylalanine

nhm

= Tetrahydronorhamane-3-carboxylic acid

Iq3

= Tetrahydroisoquinolin- (1,2,3,4) -3-carboxylic acid

ppd

= 4-phenylpiperidine-4-carboxylic acid

Tea

= Thienylalanine

pNP

= Paranitrophenylalanine

SEQUENCE LISTING

Claims (46)

A compound of the formula (I) Y ¹ - (NH-X ¹ -C =O) - (NH-X ² -C =O) - (NH-X ³ -C =O) - (NH-X ⁴ -C = O) - (NH ³ ) X ⁵ -C = O) - (NH-X ⁶ -C =O) -Y ² (I), where Y ^{1 is} either a) a hydrogen or b) a methyl group or c) an acetyl group or d) is characterized by a backbone of a chain of 1 to 32 carbon atoms, where (NH-X ¹ -C = O) represents a or L-amino acid, in particular a) valine or b) alanine or c) leucine or d) isoleucine or e) norleucine or f) asparagine or g) glutamine or h) serine or i) threonine or j) tyrosine or k) arginine or l) lysine or m) ornithine or n) phenylalanine or o) dichlorophenylalanine or p) tetrahydronorharmane-3-carboxylic acid or q) tetrahydroisoquinoline (1,2,3,4) -3-carboxylic acid or r) 4-phenylpiperidine-4-carboxylic acid or s) thienylalanine or t) phenylglycine or u) p-nitrophenylalanine v) or is replaced by a chemical bond , where (NH-X ² -C = O) is a D- or L-amino acid, in particular a) alanine or b) valine or c) leucine or d) isoleucine or e) norleucine or f) serine or g) threonine or h) Tyrosine or i) proline or j) citrulline or k) arginine or l) lysine or m) ornithine or n) histidine or o) glutamic acid or p) aspartic acid or q) tryptophan or r) cyclohexylalanine or s) cyclohexylglycine is t) or a chemical bond is replaced, wherein (NH-X ³ -C = O) is any amino acid, in particular a) L-cyclohexylalanine or b) D-cyclohexylalanine or c) L-cyclohexylglycine or d) D-cyclohexylglycine, where (NH -X ⁴ -C = O) is a small amino acid, in particular a) L-proline or b) D-proline or c) L-azetidine-2-carboxylic acid or d) D-azetidine-2-carboxylic acid, where X ⁵ -C = O) is an aromatic amino acid, in particular a) L-tyrosine or b) D-tyrosine or c) L-phenylalanine or d) D-phenylalanine, where (NH-X ⁶ -C = O) an amino acid with basic side chain, in particular ea) L-arginine or b) D-arginine or c) L-lysine or d) D-lysine or e) L-ornithine or f) D-ornithine or g) L-homoarginine or h) D-homoarginine, where Y ² either a) an OH group (the C-terminal amino acid has a terminal carboxylic acid group) or b) an amino group (in the C-terminal amino acid, the carboxylic acid is replaced by an amide group) or c) a hydrogen (in the case of the C-terminal amino acid, the carboxylic acid is replaced by an aldehyde group) or d) 7-amido-4-methylcoumarin or (combined via the carboxylic acid group) or e) paranitroanilide (combined via the carboxylic acid group) or f) by a Connecting chain is replaced by one to 35 atoms, or a molecule shortened at the C-terminus and / or at the N-terminus by not less than one amino acid, and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition N-acetyl- (NH-X ¹ -C = O) - (NH-X ² -C = O) -L-Cha-D-Pro-D-Tyr-L-Arg amide with the meanings indicated and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition N-acetyl-D-Gln-D-His-L-Cha-D-Pro-D-Tyr-L-Arg-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition N-acetyl-D-Glu-L-Cha-D-Pro-D-Tyr-L-Arg-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition N-acetyl-D-Val-D-His-L-Cha-D-Pro-D-Tyr-L-Arg-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition N-acetyl-L-Ala-L-Cha-D-Pro-D-Tyr-L-Arg-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition N-acetyl-L-Ile-L-Arg-L-Cha-D-Pro-D-Tvr-L-Arg-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition N-acetyl-L-Tyr-L-Cit-L-Cha-D-Pro-D-Tyr-L-Arg-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition N-acetyl-L-Ser-L-Ser-L-Cha-D-Pro-D-Tyr-L-Arg-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition N-acetyl-D-Val-L-Cha-D-Pro-D-Tyr-L-Arg-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition N-acetyl-L-Trp-L-Cha-D-Pro-D-Tyr-L-Arg-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition N-acetyl-L-Ser-L-Ala-L-Cha-D-Pro-D-Tyr-L-Arg-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition N-acetyl-L-Ser-L-Arg-L-Cha-D-Pro-D-Tyr-L-Arg-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition N-acetyl-D-Lys-L-Nle-L-Cha-D-Pro-D-Tyr-L-Arg-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition around N-acetyl-D-Tyr-L-Cha-D-Pro-D-Tyr-L-Arg-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition around N-acetyl-L-Arg-L-Cha-D-Pro-D-Tyr-L-Arg-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition around N-acetyl-L-Tyr-D-Pro-L-Cha-D-Pro-D-Tyr-L-Arg-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition around N-acetyl-L-Ala-D-Cha-L-Aze-D-Tyr-L-Arg-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition around N-acetyl-L-Ala-D-Cha-L-Pro-D-Tyr-L-Har-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition around N-acetyl-L-Trp-D-Cha-L-Aze-D-Tyr-L-Har-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition around N-acetyl-L-Ala-D-Cha-L-Aze-D-Tyr-L-Har-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1 characterized by the composition around N-acetyl-D-Phe-D-Cha-L-Aze-D-Tyr-L-Har-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition around N-acetyl-L-Dcp-D-Cha-L-Aze-D-Tyr-L-Har-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition around N-acetyl-L-Nhm-D-Cha-L-Aze-D-Tyr-L-Har-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition around N-acetyl-L-Iq3-D-Cha-L-Aze-D-Tyr-L-Har-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition around N-acetyl-L-Ppd-D-Cha-L-Aze-D-Tyr-L-Har-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition around N-acetyl-L-Tea-D-Cha-L-Aze-D-Tyr-L-Har-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition around N-acetyl-L-Phg-D-Cha-L-Aze-D-Tyr-L-Har-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition around N-acetyl-L-Nle-D-Cha-L-Aze-D-Tyr-L-Har-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition around N-acetyl-L-Cha-D-Cha-L-Aze-D-Tyr-L-Har-amide and pharmaceutically acceptable salts thereof. A compound according to claim 1, characterized by the composition around N-acetyl-L-Pnp-D-Cha-L-Aze-D-Tyr-L-Har-amide and pharmaceutically acceptable salts thereof. Compound mixture, characterized by a content at least two compounds according to at least one of claims 1 to 31st A compound according to any one of claims 1 to 32, wherein the compound is present as a pharmaceutically acceptable salt with an inorganic Acid formed becomes. A compound according to claim 33, wherein the compound is present as a pharmaceutically acceptable salt, which with hydrochloric acid, bromic acid and / or another halogen acid is formed. A compound according to claim 33, wherein the compound is present as a pharmaceutically acceptable salt with sulfuric acid and / or phosphoric acid is formed. A compound according to any one of claims 1 to 32, wherein the compound is a pharmaceutically acceptable salt, that with an organic acid is formed. The compound of claim 36, wherein the compound is present as a pharmaceutically acceptable salt with acetic acid, propionic acid, malonic acid, maleic acid, citric acid, succinic acid, maleic acid, benzoic acid, fumaric acid and / or a similar one carboxylic acid is formed. Medicinal products characterized by their content to one or more compounds according to at least one of claims 1 to 37 next to usual Carriers, Aids and / or additives. Diagnostic composition characterized by their content of one or more compounds after at least one of the claims 1 to 37. Use of one or more compounds according to at least one of claims 1 to 37 for the preparation of a medicament for thrombin inhibition, inhibition of fibrin formation and / or for inhibition the formation of a coagulation plug. Use of one or more compounds according to at least one of the claims 1 to 37 for the preparation of a diagnostic composition. Use according to claim 41, characterized in that in the compound of formula (I) Y ^{2 is} 7-amido-4-methylcoumarin. Use according to claim 41, characterized in that in the compound of formula (I) Y ^{2 is} paranitroanilide. A method for thrombin inhibition of humans and Animal that claims an effective amount of a compound 1 to 37 included. A Pharmaceutical Compilation That A effective plug-inhibiting amount of a compound of claim 1 to 37 and a pharmaceutically acceptable carrier. The connections can also be the substrate. A diagnostic method that links the claim 1 to 3 7 for thrombin inhibition in humans and mammals uses.