DE19603767A1 - New 1-(4-amino-benzyl)-pyrrolidine propionic acid derivatives - Google Patents

Abstract

Pyrrolidine derivatives of formula (I), and their salts are new: R1 = OH, 1-10C alkoxy, 3-10C alkenyloxy, 1-3C alkyl-(6-10C)aryloxy or NR6R7; R2 = 1-10C alkyl, 2-10C alkenyl, 3-10C alkynyl or (CH2)m-B-(CH2)nR5 (all optionally substituted by a COR1 group, or optionally substituted by one or more halo); (6-10C)aryl-(1-3C)alkyl or (6-10C)-aryl (both optionally ring substituted by 1-2 halo, 1-4C alkoxy, NO2 or CN); or 3-8C cycloalkyl, (4-10C)cycloalkyl-(1-4C)alkyl, (5-10C)cycloalkyl-(2-4C)alkenyl or (5-10C)cycloalkyl-(2-4C)alkynyl; R3 = H, 1-8C alkyl, 3-8C cycloalkyl, (6-10C)aryl-(1-3C)alkyl, 2-6C alkenyl or 3-6C alkynyl; or R2 + R3 = 2-4C alkylene, optionally substituted by halo; R4 = H, 1-6C alkyl, (6-10C)aryl-(1-3C)alkyl, 3-10C alkenyl, COO-(1-6C)alkyl, COO-(1-6C)alkyl-(6-10C)aryl or C(=NH)NH2; R5 = H, or a group of formula (i): R6, R7 = H, 1-10C alkyl, (6-10C)aryl-(1-3C)alkyl, 1-10C alkylamino or 1-10C alkylguanidino; R8 = 6-10C aryl or (6-10C)aryl-(1-3C)alkyl (both optionally substituted by one or more COR1, halo, NO2, CN, 1-4C alkoxy or amino); or 1-6C alkyl, 3-8C cycloalkyl or (2-6C)alkenyl; R9 = NH(CH2)oNHR4, O(CH2)oNH2 or O(CH2)oNH-C(=NH)NH2; A = a single or double bond; B = O, NR3 or S; Y = a direct bond, or an amino acid; m = 1-5; n = 1-5; o = 1-10.

Description

The invention relates to non-peptidic bradykinin antagonists.

In EP-A-622 361 and US 5,438,064 are heterocyclic bradykinin Antagonists described.

The present invention describes Pyrrolidinpropionsäurederivate with bradykinin antagonist activity. It relates to compounds of the formula (I)

in which the symbols have the following meaning:

• a) R
• 1. -OH,
• 2. -O- (C₁-C₁₀) -alkyl,
• 3. -O- (C₃-C₆) alkenyl,
• 4. -O- (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
• 5. -NR⁶R⁷ means;
• b) R
• 1. (C₁-C₁₀) -alkyl,
• 2. (C₂-C₁₀) -alkenyl,
• 3. (C₃-C₁₀) -alkynyl,  
• 4. (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
• 5. (C₃-C₈) -cycloalkyl,
• 6. (C₄-C₁₀) -cycloalkyl- (C₁-C₄) -alkyl,
• 7. (C₅-C₁₀) -cycloalkyl- (C₂-C₄) -alkenyl,
• 8. (C₅-C₁₀) -cycloalkyl- (C₂-C₄) -alkynyl,
• 9. - (CH₂) _m -B- (CH₂) _n -R⁵,
• 10. (C₆-C₁₀) -aryl,
• 11. a radical as defined under b) 1st, 2nd, 3rd or 9th, with COR¹ is monosubstituted,
• 12. a radical as defined under b) 1, 2, 3 or 9, in which 1 until all H atoms are substituted by halogen, or
• 13. the radical defined under b) 4 and 10, which is at the aryl with 1 or 2 equal or different residues from the series Halogen, (C₁-C₄) alkoxy and nitro, cyano is substituted;
• c) R
• 1. hydrogen,
• 2. (C₁-C₈) -alkyl,
• 3. (C₃-C₈) -cycloalkyl,
• 4. (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
• 5. (C₂-C₆) alkenyl,
• 6. (C₃-C₆) alkynyl,
• 7. R² and R³ together are (C₂-C₄) -alkyl,
• 8. a radical as defined under c) 7. with halogen is substituted;
• d) R⁴
• 1. hydrogen,
• 2. (C₁-C₆) -alkyl,
• 3. (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
• 4. (C₃-C₁₀) alkenyl,
• e) R⁵
• 1. hydrogen,
• 2. a radical of the formula (II)
• f) R⁶ and R⁷ are the same or different
• 1. hydrogen,
• 2. (C₁-C₁₀) -alkyl,
• 3. (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
• 4. (C₁-C₁₀) -alkylamino,
• 5. (C₁-C₁₀) -alkylguanidino;
• g) R⁸
• 1. (C₆-C₁₀) -aryl,
• 2. (C₁-C₆) -alkyl,
• 3. (C₃-C₈) -cycloalkyl,
• 4. (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
• 5. (C₂-C₆) alkenyl,
• 6. a remainder defined under g) 1 and 4, with one or more several radicals from the series COR¹, halogen, nitro, cyano, (C₁-C₄) alkoxy or amino substituted;
• h) R⁹
• 1. -NH- (CH₂) _o -NHR⁴
• 2. -O- (CH₂) _o -NH₂
• i) A represents a single or double bond;
• j) B is O, NR 3 or S;
• k) Y is a direct bond or an amino acid, preferably phenylalanine, 2-chlorophenylalanine, 3-chlorophenylalanine, 4-chlorophenylalanine, 2- Fluorophenylalanine, 3-fluorophenylalanine, 4-fluorophenylalanine, tyrosine, O-methyl tyrosine, β- (2-thienyl) alanine, glycine, cyclohexylalanine, Leucine, isoleucine, valine, norleucine or phenylglycine, serine or Cysteine means;
• l) m is a number 1-5,
• m) n is a number 1-5,
• n) o is a number 1-10,

and their physiologically acceptable salts.

Alkyl, alkenyl and alkynyl may be straight-chain or branched. The same applies to derived radicals such. B. alkoxy.

By cycloalkyl are also meant by (C₁-C₈) alkyl substituted rings.

(C₆-C₁₂) -aryl is, for example, phenyl, naphthyl or biphenylyl, preferably Phenyl. Accordingly applies to derived radicals, such. B. aralkyl.

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

Among physiologically acceptable salts of compounds of the formula (I) One understands both their organic and inorganic salts, as in  Remington's Pharmaceutical Sciences (17th Edition, page 1418 (1985)) are described. Due to the physical and chemical stability and the Solubility are acidic groups including sodium, potassium, calcium and Ammonium salts are preferred; for basic groups include salts of Hydrochloric acid, sulfuric acid, phosphoric acid or of carboxylic acids or Sulfonic acids, such as. Acetic acid, citric acid, benzoic acid, maleic acid, Fumaric acid, tartaric acid and p-toluenesulfonic acid are preferred.

Preference is given to compounds of the formula I, wherein

• a) R
• 1. (C₁-C₆) -alkyl,
• 2. (C₂-C₆) alkenyl,
• 3. (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
• 4. (C₃-C₈) -cycloalkyl,
• 5. (C₃-C₆) -cycloalkyl- (C₁-C₃) -alkyl,
• 6. a rest defined under a) 3 and 5, with one or more several radicals from the series halogen, nitro, cyano, (C₁- C₄) alkoxy, COR¹ or amino substituted;
• c) R⁸
• 1. (C₆-C₁₀) -aryl,
• 2. (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
• 3. a remainder defined under c) 1. and 2., with one or two several radicals from the series halogen, nitro, cyano, (C₁- C₄) alkoxy, amino or COR¹ is substituted, and

the remaining radicals are substituted as above.

Very particular preference is given to compounds of the formula I wherein

• a) R¹ is -O- (C₁-C₁₀) -alkyl;  
• b) R
• 1. (C₆-C₁₀) -aryl-CH₂-,
• 2. (C₃-C₈) -cycloalkyl,
• 3. (C₃-C₈) -cycloalkyl-CH₂-;
• c) R
• 1. (C₃-C₅) alkenyl,
• 2. phenyl- (C₁-C₃) -alkyl,
• 3. R² and R³ together are (C₂-C₃) alkyl;
• d) R⁴
• 1. hydrogen,
• e) R⁵
• 1. hydrogen,
• 2. a radical of formula (II) means
• f) R⁸
• 1. phenyl,
• 2. benzyl;
• g) R⁹
• 1. -NH- (CH₂) _o -NHR⁴,
• 2. -O- (CH₂) _O -NH₂, means;
• h) A is a single or double bond;
• i) Y 1. a single bond,
• 2. an amino acid from the series phenylalanine, β- (2- Thienyl) alanine, O-methyltyrosine, glycine, cyclohexylalanine, Leucine, isoleucine, valine, phenylglycine, serine or cysteine means;
• j) o is a number from 1-10.

The invention further relates to a process for the preparation of compounds of the formula I, which is characterized in that

• a) a compound of the formula (III) with a compound R³-X wherein
  R³ is hydrogen, (C₁-C₈) -alkyl, (C₃-C₈) -cycloalkyl, (C₆-C₁₀) -aryl (C₁-C₃) -alkyl, (C₂-C₆) -alkenyl or (C₃-C₆) -alkynyl means and
  X represents a leaving group, such as halogen, mesylate or tosylate,
  with an organometallic base such as n-butyllithium, s-butyllithium, methyllithium or phenyllithium, sodium amide and alkali metal salts of organic nitrogen bases such as lithium diisopropylamide in an inert solvent such as THF, ether, toluene or dimethoxyethane, preferably at -78 ° C to form a compound of formula IV wherein R³ is as defined above;
• b) the compound of formula IV with an alcoholate of the formula R¹Na or R¹K, wherein
  R¹ represents -OH, -O- (C₁-C₁₀) -alkyl, O- (C₃-C₆) -alkenyl or -O- (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
  refluxing the corresponding alcohol to give a compound of formula V, wherein R¹ and R³ are as defined above;
• c₁) if in formula I R² and R³ together are (C₂-C₄) -alkyl, which may be optionally substituted by halogen, and in formula V R³ is (C₂-C₆) - alkenyl,
  the compound of formula V with J₂ or radical-generating reagents in methylene chloride or THF at room temperature to give a compound of formula VI wherein R is hydrogen or (C₁-C₃) alkyl and
  R¹ is as defined under b), reacted, and
• c₂) the resulting compound of formula VI with Raney nickel at 1 atm of H₂ and diisopropylethylamine as the base for 1 h at room temperature to give a compound of formula VII wherein R is hydrogen or (C₁-C₃) alkyl and
  R¹ is as defined under b) reacted;
• d) if in formula I R² and R³ together not (C₂-C₄) alkyl, which may optionally be substituted by halogen, a compound of formula V means wherein
  R¹-O- (C₁-C₁₀) -alkyl, -O- (C₃-C₆) -alkenyl or -O- (C₆-C₁₀) -aryl (C₁-C₃) -alkyl and
  R³ is hydrogen, (C₁-C₈) -alkyl, (C₃-C₈) -cycloalkyl, (C₆-C₁₀) -aryl (C₁-C₃) -alkyl, (C₂-C₆) -alkenyl or (C₃-C₆) -alkynyl means
  with a base, z. For example, sodium hydride, and then deprotonated with a compound R²-X, wherein
  R² is (C₁-C₁₀) -alkyl, (C₂-C₁₀) -alkenyl, (C₃-C₁₀) -alkynyl, (C₆-C₁₀) -aryl (C₁-C₃) -alkyl, (C₃-C₈) -cycloalkyl, ( C₄-C₁₀) -cycloalkyl- (C₁-C₄) -alkyl, (C₅-C₁₀) -cycloalkyl- (C₂-C₄) -alkenyl, (C₅-C₁₀) -cycloalkyl- (C₂-C₄) -alkynyl, (CH₂) _m -B- (CH₂) _n -R⁵, (C₆-C₁₀) -aryl; (C₁-C₁₀) alkyl, (C₂-C₁₀) alkenyl, (C₃-C₁₀) alkynyl or - (CH₂) _m -B- (CH₂) _n -R⁵ monosubstituted with COR¹; (C₁-C₁₀) alkyl, (C₂-C₁₀) alkenyl, (C₃-C₁₀) alkynyl or (CH₂) _m -B- (CH₂) _n -R⁵, wherein 1 to all H atoms are substituted by halogen; or (C₆-C₁₀) -aryl (C₁-C₃) -alkyl or (C₆-C₁₀) -aryl, which on aryl with 1 or 2 identical or different radicals selected from the group halogen, (C₁-C₄) alkoxy and nitro , Cyano are substituted,
  BO, NR 3 or S is,
  m is a number 1-5,
  n is a number 1-5,
  R⁵ is hydrogen or a radical of the formula II, and
  X is a suitable leaving group, e.g. As halogen, mesylate or tosylate, means
  converts to a compound of formula VIII, wherein R¹, R² and R³ are as defined under d);
• e) the compound of the formula VIII with a compound of the formula wherein
  Prot. For an amino protecting group, such. B. tert. Butoxycarbonyl, stands,
  R⁴ is hydrogen, (C₁-C₆) alkyl, (C₆-C₁₀) aryl (C₁-C₃) alkyl, (C₃-C₁₀) alkenyl, C (O) -O- (C₁-C₆) alkyl , -C (O) -O- (C₁-C₆) alkyl (C₆-C₁₀) aryl, or X is a suitable leaving group, e.g. As halogen, mesylate or tosylate, means
  with a base, z. As sodium hydride, deprotonated, and then with a compound of formula VII or VIII wherein
  R is hydrogen or (C₁-C₃) -alkyl,
  R¹ is defined as under d),
  R² is (C₁-C₁₀) -alkyl, (C₂-C₁₀) -alkenyl, (C₃-C₁₀) -alkynyl, (C₆-C₁₀) -aryl (C₁-C₃) -alkyl, (C₃-C₈) -cycloalkyl, ( C₄-C₁₀) -cycloalkyl- (C₁-C₄) -alkyl, (C₅-C₁₀) -cycloalkyl- (C₂-C₄) -alkenyl, (C₅-C₁₀) -cycloalkyl- (C₂-C₄) -alkynyl, (CH₂) _m -B- (CH₂) _n -R⁵, (C₆-C₁₀) -aryl; (C₁-C₁₀) alkyl, (C₂-C₁₀) alkenyl, (C₃-C₁₀) alkynyl or - (CH₂) _m -B- (CH₂) _n -R⁵ monosubstituted with COR¹; (C₁-C₁₀) alkyl, (C₂-C₁₀) alkenyl, (C₃-C₁₀) alkynyl or (CH₂) _m -B- (CH₂) _n -R⁵, wherein 1 to all H atoms are substituted by halogen; or (C₆-C₁₀) -aryl (C₁-C₃) -alkyl or (C₆-C₁₀) -aryl, which on aryl with 1 or 2 identical or different radicals selected from the group halogen, (C₁-C₄) alkoxy and nitro , Cyano are substituted,
  BO, NR 3 or S is,
  m is a number 1-5,
  n is a number 1-5,
  R⁵ is hydrogen or a radical of the formula II, and
  R³ is hydrogen, (C₁-C₈) -alkyl, (C₃-C₈) -cycloalkyl, (C₆-C₁₀) -aryl (C₁-C₃) -alkyl or (C₃-C₆) -alkynyl,
  converts to a compound of formula IX, wherein
  R¹, R², R³, R⁴ and Prot. Are as defined under e) and
  R² and R³ together represent (C₂-C₄) alkyl optionally substituted with halogen;
• f) the amino protecting group Prot according to known methods, for. B. with tert. Butoxycarbonyl (Boc) under acidic conditions, preferably in trifluoroacetic acid (TFA) or with HCl in dimethoxyethane, or with TFA in dimethoxyethane or Dichloromethane as a preferred solvent, cleaved;
• g) optionally the resulting compound of the formula X having a free amino group wherein R¹, R², R³ and R⁴ are as defined under e) (formula IX),
  with a compound of formula II ' wherein
  R⁸ is (C₆-C₁₀) -aryl, (C₁-C₆) -alkyl, (C₃-C₈) -cycloalkyl, (C₆-C₁₀) -aryl (C₁-C₃) -alkyl, (C₂-C₆) -alkenyl; (C₆-C₁₀) -aryl or (C₆-C₁₀) -aryl (C₁-C₃) -alkyl having one or more radicals from the series COR¹, halogen, nitro, cyano, (C₁-C₄) alkoxy or amino substituted, means
  R⁹ is -NH- (CH₂) _o -NHR⁴, -O- (CH₂) _o -NH₂ or means
  R⁴ is defined as under e),
  o is a number 1-10, and
  Y means a direct bond or an amino acid,
  Coupled according to the general methods of peptide chemistry, so as to obtain a compound of formula X ', wherein R¹, R², R³ and R⁴ are as defined under g) and R⁵ wherein R⁸, R⁹ and Y are as defined under g);
• h) optionally hydrogenating the double bond of the formulas X or X 'with sodium cyanoborohydride in lower alcohols, such as methanol, at room temperature and pH ~5 to give a compound of the formula XI, wherein R¹, R², R³, R⁴, R⁵ are as defined under g) and
  A means a single bond
• i) optionally saponifying the compounds of the formulas X, X 'or XI under acidic or basic conditions or
  with HNR⁶R⁷, in which
  R⁶ and R⁷ are the same or different and are hydrogen, (C₁-C₁₀) alkyl, (C₆-C₁₀) - aryl- (C₁-C₃) alkyl, (C₁-C₁₀) -alkylamino or (C₁-C₁₀) -alkylguanidino .
  in high boiling solvents, e.g. As xylene or DMF, to obtain the compounds of formula I; and
• k) the compounds of the formula I optionally by known methods converted into their physiologically acceptable salts.

The formation of the amide bond from fragment X and II 'takes place in that a Fragment with C-terminal free carboxyl group or its activated derivative with a corresponding fragment with N-terminal free amino group and optionally one or more in the resulting compound Protecting other functions temporarily splits off introduced protecting groups and the compounds of the formula I thus obtained, if appropriate in their physiological acceptable salt transferred.

The couplings of the amino acids of the present invention were after well-known methods of peptide chemistry, see, for. B. Houben-Weyl, Methods of organic chemistry, Volume 15/2, Stuttgart 1974, carried out.

For the prevention of side reactions or for the synthesis of special peptides are the functional groups of amino acids by appropriate Protecting groups (see, e.g., T.W.Greene, "Protective Groups in Organic Synthesis "Publisher: John Wiley, 2nd edition, 1991).

As a coupling reagent can be used all possible in the peptide synthesis Activation reagents, see, eg. B. Houben-Weyl, methods of organic Chemie, Vol. 15/2, Stuttgart 1974, but in particular Carbodiimides such. N, N'-dicyclohexylcarbodiimide, N, N'-diisopropyl carbodiimide or N-ethyl-N '- (3-dimethylaminopropyl) carbodiimide. The coupling can be directly by addition of amino acid derivative with the Activation reagent and optionally a racemization suppressive additive such. For example, 1-hydroxybenzotriazole (HOBt) (W. König, R. Geiger, Chem. Ber. 103, 708 (1970)) or 3-hydroxy-4-oxo-3,4-  dihydrobenzotriazine (HOObt) (W.Kon, R. Geiger, Chem., 103, 2054 (1970)) or the preactivation of the amino acid derivative as Symmetrical anhydride or HOBt or HOObt ester can be carried out separately and the solution of the activated species in a suitable solvent for be added coupling amine.

The coupling or activation of the amino acid derivatives with one of the above mentioned activating reagents can in dimethylformamide, N-methylpyrrolidone or methylene chloride or a mixture of the above Solvents are carried out.

The coupling of the Boc-protected diamines to the piperidine derivatives to form a urea functionality is preferably carried out using Carbodiimides, phosgene or chloroformate as activated Carbonylreagenzien.

The synthesis of the guanidino derivatives is carried out starting from the corresponding Amines with amidino-group-transferring reagents, preferably 1-H-pyrazole carboxamidine.

The compounds according to the invention have, individually or in combination bradykinin-antagonistic effect, tested in different models (Handbook of Exp. Pharmacol. Vol. 25, Springer Verlag, 1970, Pp. 53-55), such as On isolated rat uteri, on guinea pigile ileum or at the isolated guinea pig pulmonary artery.

The measurement of binding to the bradykinin B₂ receptor from Guinea pig ileum is described below (R.B. Innis et al., Proc.Natl.Acad.Sci.USA; 17 (1981) 2630:

1. Ligand: 3H-BRADYKININE (from NEN Du Pont)  

2. Buffer approaches:
a) TES buffer:
25 mM TES (Sigma, Order No .: T-4152)
1 mM 1,10-phenanthroline (SIGMA; Order No .: P-9375)
b) incubation buffer:
25mM TES (SIGMA; Order No .: T-4152)
1 mM 1,10-phenanthroline (SIGMA; Order No .: P-9375)
0.1% albumin, bovine (SIGMA; Order No .: A-7906)
140 μg / ml Bacitracin (SIGMA; Order No .: B-0125)
1 mM dithiothreitol (SIGMA; Order No .: D-0632)
1 μM captopril → 1 - [(2S) -3-mercapto-2-methylpropionyl] -L-proline
Both buffers are adjusted to pH 6.8 with 5 molar NaOH.

3. Membrane preparation:
Guinea-pig ilea are roughly cleared of the intestinal contents by gentle brushing and cleaned in 0.9% NaCl solution.

The 2 cm long pieces of Ilea are transferred into ice-cold TES buffer (about 1 g / 10 ml) and homogenized with the utraturrax for about 30 sec. In an ice bath. The Homogenate is then filtered through 3 layers of gauze and the filtrate at Centrifuged 50,000 g / 10 minutes.

The supernatant is discarded, the pellet in the same volume of TES buffer rehomogenized and again centrifuged at 50,000 g / 10 minutes.

The pellet is rehomogenized in incubation buffer (about 1g / 5 ml) and in Cryotube, 2 ml portioned, frozen at -70 ° C.

The protein concentration of the finished membrane suspension becomes LOWRY determined and should be about 15 ug / 100 ul.

4. Binding test:
All incubations are carried out at room temperature for 60 minutes on microtiter plates (96 × 300 μl) in 200 μl volume. All batches in incubation buffer. For this purpose, 50 μl of the radioligand, 50 μl of the test specimen and 100 μl of the membrane suspension are successively pipetted into the wells of the microtiter plate.

a) Saturation experiments (hot saturation):
Production of the ³H-bradykinin solution: For the saturation experiments, the concentrations 0.05, 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 1.5, 2.0, 2.5 and 3.0 nmol / l, which corresponds to 0.05 to 3.0 pmol / ml, are used. After the preparation of the appropriate dilutions, 50 μl per sample are presented.

Non-specific binding: For each concentration of radioactive ligand must the non-specific binding can be determined. This can be done by adding a high concentration (1-100 μmol) of unlabeled ligand, other Antagonists or agonists of the bradykinin receptor can be achieved. In Hoe 140 (10 μmol / l) is used in this test. This will be 1.862 mg in 1 ml of dimethyl sulfoxide (DMSO) dissolved, diluted 1:25 with incubation buffer and from this solution add 50 μl to the samples in the microtiter plate. The Reaction is started by the addition of 100 μl of the membrane suspension.

b) Competition Experiments (IC₅₀):
Here, a fixed size of the radioactive ligand (0.25 to 0.3 nmol / l3H-bradykinin) and various concentrations of unlabelled agonists or antagonists are used. To each 50 .mu.l of the ³H-bradykinin solution, 50 .mu.l of the preparations or standards to be tested are added in the concentrations 10 ^-5 to 10 ^-10 mol / l and the reaction started by adding 100 .mu.l of membrane suspension. Also in this test, 3-fold determinations are performed and three samples are incubated with 10 μMol / l Hoe 140 to determine nonspecific binding.

The products to be tested for competition are always in one Concentration of 1 mmol / l dissolved in dimethyl sulfoxide (DMSO), and then further diluted with DMSO. This solution will then be 1:25 with Diluted incubation buffer.

After incubation, the samples are transferred to the Skatron cell harvester via a previously impregnated with 0.1% PEI (polyethyleneimine) Whatmann GF / B Filtered paper strips and with, per sample, 10 ml of ice-cold TES buffer rewashed. The still wet filters are placed in mini-scintillation vials punched out and filled with 3 ml scintillator. After approx. 12 hours soaking time, the samples are shaken briefly and placed in the Beta counter measured.

c) Screening:
In primary screening, only 1-2 concentrations of the investigational medicinal product (10 ^-5 and 10 ^-6 mol / l) are generally used. If a displacement of the radioligand of 50% or more is detectable at the highest concentration, a complete analysis (competition experiment) with at least 8 concentrations is carried out.

4. Evaluation:
Evaluation is carried out by means of the LIGAND program package (Mc Pherrson, Minson & Rodbard, Marketing: Elsevier-BIOSOFT), which performs the necessary calculations to determine IC₅₀ and _Ki values. This program also performs graphical representations of the saturation and displacement curves as well as the SCATCHARD plot, HILL plot or HOFSTEE plot.

The determination of the antagonistic effect on the bradykinin-induced contraction of the guinea pig ileum is carried out according to the following protocol:
Guinea pigs weighing about 300 g (Morioth strain, ♂, ♀) are killed by stroke of the neck and bled to death. The ileum is prepared in a length of about 20 cm, rinsed with Tyrode solution (Recordspritze) and thus freed from intestinal contents. Now it is divided into 1.5 cm long sections. These are fixed in 10 ml organ baths filled with Tyrode solution and connected with strain gauges (isometric contraction measurement). The preload is 1g. The Tyrode solution is heated in a water bath to 37 ° C and bubbled with compressed air.

After an interval of 30 min. is started with the attempt. After recording the biological zero line, bradykinin in a final concentration of 4 × 10 ^-8 mol / l is added per organ bath and the concentration is recorded. After that, 3 min. rinsed with Tyrode solution and after a rest period of 20 min. Bradykinin added again. The maximum of the contraction is reached (control). Rinse again, rest. Now the bradykinin antagonist is added (exposure time 10 min.). Bradykinin is then added again and the contraction now compared with that of the control compared. The test is recorded on a pen.

Tyrode solution (mM):

NaCl 137 glucose 5.05 KCl 2.68 NaHCO₃ 11.9 NaH₂PO₄ 0.47 MgCl₂ × 2 H₂O 0.49 CaCl₂ × 2 H₂O 0.68

Amplifier: TF6 V3 Fa. Fleck, Mainz
Ink pen: Goerz Metrawatt SE 460, BBC
Bradykinin: Fa. Bachem

Table 1*

Biological data

For oral administration or for application to the mucous membranes the active compounds with the customary additives such as carriers, Stabilizers or inert diluents and mixed by conventional Methods into suitable dosage forms, such as tablets, dragees, Capsules, aqueous, alcoholic or oily suspensions or aqueous, alcoholic or oily solutions. As inert carrier can z. Gum arabic, Magnesia, magnesium carbonate, potassium phosphate, lactose, glucose,  Magnesium stearyl fumarate or starch, especially corn starch used become. The preparation may be both dry and wet granules respectively. As oily carriers or solvents, for example vegetable or animal oils, such as sunflower oil and cod liver oil.

A preparation for topical application may be an aqueous or oily solution, Lotion, emulsion or jelly, ointment or ointment or, if possible, in Spray form, optionally with the addition of a polymer Liability can be improved.

For the intranasal form of application, the compounds with the conventional additives such as stabilizers or inert diluents mixed and brought by conventional methods into suitable dosage forms, such as aqueous, alcoholic or oily suspensions or aqueous, alcoholic or oily solutions. Aqueous intranasal preparations may include chelating agents, Ethylenediamine-N, N, N ', N'-tetraacetic acid, citric acid, tartaric acid or their Salts are added. The application of the nasal solutions can by means of Dosing or done as a nasal drops with viscosity-increasing proportion or nasal gels or nose creams.

The therapeutic benefit of the compounds of the invention includes all pathological conditions, the bradykinin and bradykinin-related peptides mediated, triggered or supported. This includes u. a. Traumas, like Wounds, burns, rashes, erythema, edema, angina, arthritis, Asthma, allergies, rhinitis, shock, inflammation, pancreatitis, arteriosclerosis, Virus disorders, low blood pressure, pain, itching and altered Sperm motility.

The invention therefore also relates to the use of compounds Formula I as a remedy and pharmaceutical preparations containing these compounds contain.  

Pharmaceutical preparations contain an effective amount of the active ingredient of Formula I - individually or in combination - together with an inorganic or organic pharmaceutically acceptable carrier.

The application can be enteral, parenteral - such. B. subcutaneously, i. m. or I. v. - sublingual, epicutaneous, nasal, rectal, intravaginal, intrabuccal or by inhalation respectively. The dosage of the drug depends on the warm-blooded species, the Body weight, age and type of application.

The pharmaceutical preparations of the present invention are in themselves known solution, mixing, granulation or coating process.

For inhalative use nebulizers or aerosols under Use of inert carrier gases are used.

For intravenous, subcutaneous, epicutaneous or intradermal administration the active compounds or their physiologically acceptable salts, if desired, with the pharmaceutically customary auxiliaries, for example for Isotonization or pH adjustment and solubilizers, emulsifiers, or other excipients, in solution, suspension or emulsion.

Due to the short half lives of some of the drugs described in Body fluids, the use of injectable sustained release preparations makes sense. As dosage forms z. As oily crystal suspensions, microcapsules, Rods or Implants are used, the latter being made of tissue-compatible Polymers, in particular biodegradable polymers, such as z. B. based on polylactic acid-polyglycolic acid copolymers or Human albumin can be constructed.

A suitable dose range for topical and inhalational uses are Solutions with 0.01-5 mg / ml, in systemic administration forms are 0.01-10  mg / kg suitable.

The abbreviations used for amino acids correspond to the usual three-letter code in peptide chemistry as described in Europ. J. Biochem. 138, 9 (1984). Further abbreviations used are listed below:
Cha: cyclohexylalanine
Nle: norleucine
Oic: 2S, 3aS, 7aS-hexahydro-2-indolinyl-carboxylic acid
Phg. phenylglycine
Ser (t-But): O-tert-butyl-L-serine
D-Tic R-1,2,3,4-tetrahydro-3-isoquinolylcarboxylic acid
Thi: L-β (2-thienyl) alanine
Boc: t-butyloxycarbonyl
CH₂Cl₂: dichloromethane
Cl: Chemical ionization
DCC: N, N-dicyclohexylcarbodiimide
DCI: Desorption-Chemical Ionization
DIP: diisopropyl ether
DME: dimethoxyethane
DMF: N, N-dimethylformamide
EA: ethyl acetate
ESI: Electron Spray Ionization
FAB: Fast Atom Bombardment
nH: n-heptane
HOBt: 1-hydroxybenzotriazole
MeOH: methanol
MTB: methyl tert-butyl ether
RT: room temperature
THF: tetrahydrofuran
Totu: O- [cyano (ethoxycarbonyl) methyleneamino] -1,1,3,3-tetramethyluronium tetrafluoroborate
The invention is illustrated by the following examples.

example 1 2- (1- (4-t-Butyloxycarbonylaminomethylbenzyl) -pyrrolidin-2-yl) -3-naph-Thalin-2-yl propionate a) 2- (pyrrolidin-2-ylidene) -3- (naphthyl-2-yl) -propionic acid ethyl ester

To pyrrolidin-2-ylidene-acetic acid ethyl ester (18 g, 116 mmol) in DME (500 mL) at 0 ° C under argon was added sodium hydride (4.5 g of a 60% suspension in mineral oil). After completion of the addition, the cooling was removed and after 40 min. was slowly added dropwise at RT 2-bromomethylnaphthalene (228 g, 103 mmol). After 2 h, H₂O (300 ml) was added and concentrated in vacuo. The residue is adjusted to pH = 7 with 4N HCl and extracted 3 times with ethyl acetate, dried (MgSO₄) and concentrated. Chromatography on SiO with EE / H 1/2 as the eluent gave the title compound as an oil.
R _f (EE / nH 1/1) = 0.5.
MS (DCI) = 296 (M + H).

b) 4-t-butyloxycarbonylaminomethylbenzyl alcohol

4-Aminomethylbenzoic acid (24 g, 159 mmol) was added under argon to a suspension of sodium borohydride (15 g, 428 mmol) in DME (800 mL). Now a solution of iodine (40.6 g, 159 mmol) in DME (150 ml) was added dropwise at 6-10 ° C, the solution foaming strongly. After completion of the addition was allowed to act for 2 hours without cooling, then was refluxed for 18 h. After cooling to RT, methanol (100 ml) was added dropwise, the suspension initially foaming vigorously. Concentrate in vacuo and add 20% aqueous KOH (300 mL). After 18 h at rt, it was extracted 3 times with MTB (3 x 300 mL). The organic phases were washed once with saturated NaCl solution, dried (MgSO₄) and concentrated in vacuo. To the thus-obtained alcohol (14 g, 102 mmol) in THF (150 mL) were added diisopropylethylamine (17.5 mL, 107 mmol) and di-t-butyl pyrocarbonate (22 g, 107 mmol) dissolved in THF (100 mL). , added at RT. After 18 h, it was concentrated in vacuo and taken up in ethyl acetate. The organic phase was washed once each with 5% NaHSO₄-, saturated Na₂CO₃- and saturated NaCl solution, dried over MgSO₄ and concentrated in vacuo. The residue was stirred with n-heptane (100 ml) and suction filtered and washed with n-heptane (200 ml) to give the title compound as a colorless powder.
R _f (EE / nH 2/1) = 0.45.
¹H-NMR (CDCl₃) δ = 7.25 (m, 4H), 4.8 (s (broad), 1H), 4.65 (m, 2H), 4.3 (d, J = 6.0Hz, 2H), 1.25 (s, 9H) ,

c) (4-t-Butyloxycarbonylaminomethylbenzyl) -methanesulfonate

To title compound 1b (10 g, 30 mmol) and triethylamine (8.7 mL, 63 mmol) in THF (100 mL) was added dropwise mesyl chloride (5.0 mL, 50 mmol) at 0 ° C. After 1 h, a pH = 7 was set and concentrated in vacuo. Now ethyl acetate (300 ml) was added and washed 1 × with 5% NaHSO₄-, saturated NaHCO₃- and NaCl solution, dried (MgSO₄) and concentrated again to give the title compound as an oil.
R _f (EE / nH 2/1) = 0.6.
¹H-NMR (CDCl₃) δ = 7.35 (m, 4H), 5.2 (s, 2H), 4.85 (s wide, 1H), 4.3 (d, J = 6Hz, 2H), 2.9 (s, 3H), 1.2 ( s, 9H).

d) 2- (1- (4-t-Butyloxycarbonylaminomethylbenzyl) -pyrrolidin-2-ylidene) -3-- naphthyl-2-yl-propionic acid ethyl ester

To the title compound of Example 1a (9.8 g, 33 mmol) in DMF (150 ml) was added sodium hydride (2.7 g, 60% in mineral oil) at 5 ° C. After completion of the addition was 30 min. stirred without cooling and the title compound 1c (10.5 g, 33 mmol) dissolved in DMF (10 ml) was added dropwise. After 2 h at RT H₂O (20 ml) was added and concentrated in vacuo. To the residue was added H₂O (200 ml) and extracted 3 times with 200 ml of ethyl acetate. The organic extracts were dried (MgSO₄) and concentrated in vacuo. Chromatography on SiO with EE / H 1/1 as eluent afforded the title compound as an oil.
R _f (EE / nH 1/1) = 0.4. MS (DCI) = 515 (M + H).

e) 2- (1- (4-t-Butyloxycarbonylaminomethylbenzyl) -pyrrolidin-2-yl) -3-na-phthalin- 2-yl-propionic acid ethyl ester

To the title compound according to Example 1d (200 mg, 0.39 mmol) in methanol (6 ml) was added at room temperature sodium cyanoborohydride (25 mg, 0.39 mmol) and a spatula tip of bromocresol green. With 0.1 N HCl, the pH was now adjusted to its transition point. After 18 h, it was concentrated and H₂O (20 ml) was added. The pH was adjusted to 10 and extracted 2x with MTB (100 ml). The organic phase was dried (MgSO₄) and concentrated in vacuo to give the title compound as an oil.
R _f (EE / MeOH 5/1) = 0.2.
M (DCI) = 517 (M + H).

Example 2 2- (1- (4-Aminomethyl-benzyl) -pyrrolidine-2-ylidene) -3-naphthyl-2-yl propionic acid ethyl ester · trifluoroacetate

To the title compound of Example 1d (14 mg, 1 mmol) in dichloromethane (10 ml) was added at 0 ° C trifluoroacetic acid (10 ml). After 4 h, it was concentrated and digested twice with DIP (10 ml) and lyophilized to give the title compound as an amorphous powder.
R _f (CH₂Cl₂ / MeOH / CH₃CO₂H / H₂O 130/30/5/5) = 0.3.
MS (ESI) = 415 (M + H).

Example 3 2- (1- (4-Aminomethyl-benzyl) -pyrrolidin-2-yl) -3-naphthalen-2-yl- Propionic acid ethyl ester · di-trifluoroacetate

The title compound according to Example 1 (516 mg, 1 mmol) was reacted analogously to Example 2 to give the title compound as an amorphous powder.
R _f (CH₂Cl₂ / MeOH / CH₃CO₂H / H₂O 130/30/5/5) = 0.15.
MS (ESI) = 417 (M + H).

Example 4 2- (1- (4-Guanidinomethylbenzyl) -pyrrolidine-2-ylidene) -3-naphthalen-2-y l propionate

To the title compound of Example 2 (528 mg, 1 mmol) in DMF (20 mL) and diisopropylethylamine (0.44 mL, 2.6 mmol) at RT was 1H-pyrazole-1-carboxamidine hydrochloride (150 mg, 1.0 mmol). added. After 18 hours, the product was precipitated by the addition of DIP (100 ml).
R _f (CH₂Cl₂ / MeOH / CH₃CO₂H / H₂O 130/30/5/5) = 0.4.
MS (ESI) = 457 (M + H).

Example 5 2- (1- (4-Guaninomethylbenzyl) -pyrrolidin-2-yl) -3-naphthalen-2-yl- propionate

The title compound according to Example 3 (530 mg, 1 mmol) was reacted analogously to the title compound according to Example 4 to give the title compound according to Example 5.
R _f (CH₂Cl₂ / MeOH / CH₃CO₂H / H₂O 130/30/5/5) = 0.2.
MS (ESI) = 459 (M + H).

Example 6 2- (1- (4 - ((1- (8-Aminooctylcarbamoyl) -4-phenyl-piperidine-4-carbonyl) -L-β- (2- thienyl) alaninyl) -aminomethylbenzyl) -pyrrolidine-2-ylidene) -3-naphthal-in-2-yl propionic acid ethyl ester · trifluoroacetate a) 4-phenylpiperidine-4-carboxylic acid methyl ester

4-Phenylpiperidine-4-carboxylic acid tosylate (200 g, 0.53 mol) was boiled in methanol (1000 mL) with addition of thionyl chloride (150 mL). After completion of HCl evolution, the mixture was concentrated to saturate with saturated Na₂CO₃ solution 11 and extracted 3 times with ethyl acetate (3 × 300 ml) After drying the organic phases over MgSO₄, the mixture was concentrated in vacuo to give the product as an oil.
R _f (EE / MeOH 10/1) = 0.4.
MS (DCI) = 220 (M + H).

b) 1- (8-tert-Butyloxycarbonylaminooctylcarbamoyl) -4-phenylpiperidine-⁴- carboxylic acid methylester

To 8-Boc-aminooctylamine hydrochloride (10g, 35mmol) and diisopropylethylamine (4.6g, 35mmol) in DMF (100ml) at RT was added N, N'-carbonyldiimidazole (5.8g, 35mmol) , After 4 h, the compound according to Example 1a (7.8 g, 35 mmol) was added and the mixture was stirred at RT for a further 18 h. Then ethyl acetate (500 ml) was added and washed 1 time (100 ml) with 5% NaHSO₄, saturated Na₂CO₃ and saturated NaCl solution. The organic phase was dried over MgSO₄ and concentrated in vacuo.
Yield compound 1b) 15 g.
R _f (EE) = 0.6.
MS (FAB) = 490 (M + H).

c) 1- (8-tert-Butyloxycarbonylaminooctylcarbamoyl) -4-phenylpiperidine-4- carboxylic acid

The title compound according to Example 1b (15 g, 31 mmol) was stirred in MeOH / H₂O 2/1 (300 ml) and sodium hydroxide (4.8 g, 120 mmol) at RT for 18 h. It was then concentrated under reduced pressure and adjusted to pH = 3 with 1N HCl. Now was extracted 3 × with 100 ml of ethyl acetate. The combined organic extracts were dried over MgSO₄. The title compound was obtained by concentration in vacuo.
MS (DCI) = 476 (M + H).

d) 1- (8-tert-Butyloxycarbonylaminooctylcarbamoyl) -4-phenylpiperidine-4- carbonyl-β (2-thienyl) alanine-methylester

To the compound of Example 1c (14 g, 29 mmol) in DMF (100 mL) at RT was added HOBt (6 g, 44 mmol) and DCC (7.2 g, 35 mmol). After 30 min. β (2-thienyl) alanine methyl ester hydrochloride (6.1 g, 29 mmol) and triethylamine (3 g, 29 mmol) were added. After 18 h, filtered and the filtrate diluted with ethyl acetate (500 mL). The organic phase was washed 1 time (100 ml) with 5% NaHSO₄, saturated Na₂CO₃ and saturated NaCl solution, dried over MgSO₄ and concentrated in vacuo. The title compound was obtained after chromatography on SiO₂ with MTB as the eluent.
R _f (MTB) = 0.4.
MS (FAB) = 643 (M + H).

e) 1- (8-tert-Butyloxycarbonylaminooctylcarbamoyl) -4-phenylpiperidine-4- carbonyl-β- (2-thienyl) alanine

The title compound according to Example 1d (12.8 g, 27 mmol) in MeOH / H₂O 2/1 (300 ml) and sodium hydroxide (4.4 g, 110 mmol) were reacted analogously to Example 1c to obtain the title compound.
MS (FAB) = 629 (M + H).

f) 2- (1- (4 - ((1- (8-Butyloxycarbonylaminooctylcarbamoyl) -4-phenyl-piperid-in-4 carbonyl) -L-β- (2-thienyl) alaninyl) -aminomethylbenzyl) -pyrrolidine-2-ylidene) -3- naphthalene-2-yl-propionic acid ethyl ester

To the title compound according to Example 6e (200 mg, 0.32 mmol) in DMF (5 ml) was added HOBt (65 mg, 0.48 mmol) and DCC (72 mg, 0.35 mmol) at RT. After 1 h, the title compound according to Example 2 (170 mg, 0.31 mmol) and triethylamine (43 mL, 0.31 mmol) were added. After 18 h, the mixture was concentrated in vacuo. To the residue was added acetonitrile and filtered. The filtrate was again concentrated. After addition of ethyl acetate (50 ml) was washed 1 × with saturated Na₂CO₃- and NaCl solution, dried (MgSO₄) and concentrated in vacuo. Chromatography on SiO gave the title compound (eluent EE).
R _f (EE) = 0.45.
MS (ESI) = 1026 (M + H).

g) 2- (1- (4 - ((1- (8-aminooctylcarbamoyl) -4-phenyl-piperidine-4-carbonyl) -L-β- (2- thienyl) alaninyl) -aminomethylbenzyl) -pyrrolidine-2-ylidene) -3-naphthal-in-2-yl propionic acid ethyl ester · trifluoroacetate

The title compound according to Example 6f (150 mg, 0.15 mmol) was reacted analogously to Example 2, whereby the title compound according to Example 6 was obtained as an oil.
R _f (CH₂Cl₂ / MeOH / CH₃CO₂H / H₂O 130/30/5/5) = 0.35.
MS (ESI) = 925 (M + H).

Example 7 2- (1- (4 - ((1- (8-Aminooctylcarbamoyl) -4-phenyl-piperidine-4-carbonyl) -L-β- (2- thienyl) alaninyl) -aminomethylbenzyl) -pyrrolidin-2-yl) -3-naphthalen-2 - yl- propionate

The title compound was obtained by reacting the title compound according to Example 6 (120 mg, 0, 13 mmol) analogously to Example 1e.
R _f (CH₂Cl₂ / MeOH / CH₃CO₂H / H₂O 130/30/5/5) = 0.3.
MS (FAB) = 927 (M + H).

Example 8 2- (1- (4- (1- (8-t-Butyloxycarbonylaminooctylcarbamoyl) -4-phenyl-piperi-din-4- carbonyl) -aminomethylbenzyl) -pyrrolidine-2-ylidene) -3-naphthalen-2-yl- propionate

The title compound was prepared analogously to Example 6d from the title compound according to Example 2 (170 mg, 0.32 mmol) and the title compound according to Example 6c (150 mg, 0.32 mmol).

R _f (EE) = 0.3 MS (ESI) = 872 (M + H).

Example 9 2- (1- (4- (1- (8-t-Butyloxycarbonylaminooctylcarbamoyl) -4-phenyl-piperi-din-4- carbonyl) -aminomethylbenzyl) -pyrrolidin-2-yl) -3-naphthalen-2-yl- propionate

The title compound was obtained from the title compound according to Example 8 (285 mg, 0.33 mmol) analogously to the reaction according to Example 1e.
R _f (EE / MeOH 1/1) = 0.3
MS (FAB) = 874 (M + H).

Example 10 2- (1- (4- (1- (8-Aminooctylcarbamoyl) -4-phenyl-piperidine-4-carbonyl) - aminomethylbenzyl) -pyrrolidin-2-ylidene) -3-naphthyl-2-yl-pragnp. acid ethyl ester. trifluoroacetate

The title compound was obtained analogously to the reaction according to Example 2 from the title compound according to Example 8 (120 mg, 0.14 mmol).
R _f (CH₂Cl₂ / MeOH / CH₃CO₂H / H₂O 130/20/5/5) = 0.5.
MS (FAB) = 772 (M + H).

Example 11 2- (1- (4- (1- (8-Aminooctylcarbamoyl) -4-phenyl-piperidine-4-carbonyl) - aminomethylbenzyl) -pyrrolidin-2-yl) -3-naphthyl-2-yl-propionic acid ethyl ester · Di trifluoroacetate

The title compound was obtained analogously to the reaction according to Example 2 from the title compound according to Example 9 (120 mg, 0.14 mmol).
R _f (CH₂Cl₂ / MeOH / CH₃CO₂H / H₂O 130/30/5/5) = 0.3.
MS (FAB) = 774 (M + H).

Example 12 2- (1- (4- (1- (8-Guanidinooctylcarbamoyl) -4-phenyl-piperidine-4-carbonyl -) - aminomethylbenzyl) -pyrrolidin-2-ylidene) -3-naphthyl-2-yl-propionic acid ethyl ester. trifluoroacetate

The title compound was obtained analogously to the reaction according to Example 4 from the title compound according to Example 10 (120 mg, 0.14 mmol).
R _f (CH₂Cl₂ / MeOH / CH₃CO₂H / H₂O 130/20/5/5) = 0.5.
MS (FAB) = 814 (M + H).

Example 13 2- (1- (4- (1- (8-Guanidinooctylcarbamoyl) -4-phenyl-piperidine-4-carbonyl -) - aminomethylbenzyl) -pyrrolidin-2-yl) -3-naphthyl-2-yl-propionic acid ethyl ester · trifluoroacetate

The title compound was obtained analogously to the reaction according to Example 4 from the title compound according to Example 11 (120 mg, 0.14 mmol).
R _f (CH₂Cl₂ / MeOH / CH₃CO₂H / H₂O 130/20/5/5) = 0.25.
MS (FAB) = 816 (M + H).

Example 14 2- (1- (4- (1- (6-Aminohexylcarbamoyl) -4-phenyl-piperidine-4-carbonyl) - aminomethylbenzyl) -pyrrolidin-2-ylidene) -3-naphthyl-2-yl-pragnp. acid ethyl ester. trifluoroacetate

The title compound was prepared analogously to the reaction according to Examples 6d and 2 from the title compound according to Example 2 (350 mg, 0.67 mmol) and 1- (6-t-butyloxycarbonylaminohexylcarbamoyl) -4-phenyl-piperidine-4-carboxylic acid-e ( 300 mg, 0.67 mmol).
R _f (CH₂Cl₂ / MeOH / CH₃CO₂H / H₂O 130/30/5/5) = 0.5.
MS (FAB) = 744 (M + H).

Example 15 2- (1- (4- (1- (6-Aminohexylcarbamoyl) -4-phenyl-piperidine-4-carbonyl) - aminomethylbenzyl) -pyrrolidin-2-yl) -3-naphthyl-2-yl-propionsäureethy-lester

The title compound was obtained analogously to the reaction according to Example 1e from the title compound of Example 14 (120 mg, 0.14 mmol).
R _f (CH₂Cl₂ / MeOH / CH₃CO₂H / H₂O 130/20/5/5) = 0.25.
MS (FAB) = 776 (M + H).

Example 16 2- (1- (4- (1- (6-Guanidinohexylcarbamoyl) -4-phenyl-piperidine-4-carbonyl -) - aminomethylbenzyl) -pyrrolidin-2-yl) -3-naphthyl-2-yl-propionsäureethy-lester

The title compound was obtained analogously to the reaction according to Example 4 from the title compound according to Example 15 (250 mg, 0.29 mmol).
R _f (CH₂Cl₂ / MeOH / CH₃CO₂H / H₂O 130/30/5/5) = 0.1,
MS (ESI) = 788 (M + H).

Example 17 1- (4-aminomethylbenzyl) -2,3,3a, 4,5,6-hexahydroindole-7-carboxylic acid etyl ester · trifluoroacetate a) 2,2-Dimethyl-5- (3-allyl-pyrrolidin-2-ylidene) - [1,3] -dioxan-4,6-dione

To 2,2-dimethyl-5-pyrolidin-2-ylidene- (1,3-dioxane-4,6-dione (3g, 13.5 mmol) in dry THF (120 ml) was added under argon atmosphere at -65 ° ° C n-butyllithium (24 ml of a 15% solution in hexane) was added and then the cooling was removed and after 45 min then cooled again to -65 ° C. To this suspension was added allyl bromide (1.2 ml, 13.9 After 30 minutes, 30 ml of 10% HCl solution was added and extracted 3 times with ethyl acetate (3 × 100 ml) The combined organic extracts were washed once with saturated NaCl solution (100 ml), dried ( MgSO₄) and concentrated in vacuo to give the title compound as colorless amorphous crystals.
R _f (EE / nH 2/1) = 0.5. MS (CI) = 251 (M⁺)

b) 3-Allyl-pyrrolidin-2-ylidene-acetic acid ethyl ester

The title compound of Example 17a (3.2 g, 12.7 mmol) in ethanol (60 mL, absolute) and sodium ethylate (16.5 mL of a 1 M solution in absolute ethanol) was refluxed for 18 hours. It was then concentrated, added H₂O (30 ml) and adjusted with 2 N HCl to pH = 5-6. Now it was extracted twice with ethyl acetate (100 ml), the organic extracts were dried (MgSO₄) and concentrated in vacuo. The title compound fell as an oil.
R _f (EE / nH 2/1) = 0.6
MS (CI) = 195 (M⁺),

c) 5-iodo-2,3,3a, 4,5,6-hexahydro-1H-indole-7-carboxylic acid ethyl ester

To the title compound of Example 17b (5, 1g, 23 mmol) in dichloromethane (100 ml) was added dropwise under argon at -5 ° C a solution of N-iodosuccinimide (6.9 g, 30 mmol) in dichloromethane (100 ml). After 3 hours (during which time (3-allylpyrrolidin-2-ylidene) -2-iodoacetic acid ethyl ester formed) cooling was removed. After 18 h at RT was washed 1 time with half-saturated Na₂SO₃ solution (100 ml) and 1 time with H₂O (100 ml). The organic phase was dried (CaCl₂) and concentrated in vacuo. The crystalline residue was crystallized from ethanol.
R _f (EE / nH 1/2) = 0.25
MS (ESI) = 322 (M + H)

d) 2,3,3a, 4,5,6-hexahydro-1H-indole-7-carboxylic acid ethyl ester

The title compound of Example 17c (500 mg, 1.56 mmol) and Raney Nickel (100 mg) were dissolved in a mixture of methanol (5 mL), THF (10 mL) and diisopropylethylamine (2 mL) in a hydrogen atmosphere (1 atm). Stirred for 1 h. The mixture was then filtered off from the catalyst over kieselguhr, saturated NaCl solution (50 ml) was added and the mixture was extracted twice with ethyl acetate. The organic extracts are dried (MgSO₄) and concentrated in vacuo to afford the title compound was crystalline.
R _f (DIP) = 0.6 MS (El) = 196 (M + H),
Mp .: 74 ° C (crystallized from DIP).

e) 1- (4-t-Butyloxycarbonylaminomethylbenzyl) -2,3,3a, 4,5,6-hexahydroindoline 7-carboxylic acid ethyl ester

The title compound was obtained analogously to the reaction according to Example 1d from the title compound according to Example 1c (1.05 g, 3.3 mmol) and the title compound according to Example 17d (640 mg, 3.3 mmol).
R _f (E / nH 1/4) = 0.1.
MS (ESI) = 415 (M + H).

f) 1- (4-aminomethylbenzyl) -2,3,3a, 4,5,6-hexahydroindole-7-carboxylic acid ethyl ester · trifluoroacetate

The title compound according to Example 17 was obtained analogously to the reaction according to Example 2 from the title compound according to Example 17e (200 mg, 0.48 mmol).
R _f (CH₂Cl₂ / MeOH / CH₃CO₂H / H₂O 130/30/5/5) = 0.3.
MS (ESI) = 315 (M + H).

Example 18 Ethyl 1 (4-aminomethylbenzyl) -octahydro-indole-7-carboxylate · trifluoroacetate

The title compound was obtained from the title compound according to Example 17e (414 mg, 1 mmol) analogously to the reaction according to Examples 1e and 2.
R _f (CH₂Cl₂ / MeOH / CH₃CO₂H / H₂O 130/30/5/5) = 0.4,
MS (ESI) = 317 (M + H).

Claims (6)

1. Compounds of the formula (1) in which the symbols have the following meaning:

• a) R¹
• 1. -OH,
• 2. -O- (C₁-C₁₀) -alkyl,
• 3. -O- (C₃-C₆) alkenyl,
• 4. -O- (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
• 5. -NR⁶R⁷ means;
• b) R²
• 1. (C₁-C₁₀) -alkyl,
• 2. (C₂-C₁₀) -alkenyl,
• 3. (C₃-C₁₀) -alkynyl,
• 4. (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
• 5. (C₃-C₈) -cycloalkyl,
• 6. (C₄-C₁₀) -cycloalkyl- (C₁-C₄) -alkyl,
• 7. (C₅-C₁₀) -cycloalkyl- (C₂-C₄) -alkenyl,
• 8. (C₅-C₁₀) -cycloalkyl- (C₂-C₄) -alkynyl,
• 9. - (CH₂) _m -B- (CH₂) _n -R⁵,
• 10. (C₆-C₁₀) -aryl,
• 11. a radical as defined under b) 1, 2, 3 or 9, monosubstituted with COR¹,
• 12. a radical as defined under b) 1, 2, 3 or 9, in which 1 to all H atoms are substituted by halogen, or
• 13. the b) 4 and 10 defined radical which is substituted on the aryl with 1 or 2 identical or different radicals from the series halogen, (C₁-C₄) alkoxy and nitro, cyano;
• c) R³
• 1. hydrogen,
• 2. (C₁-C₈) -alkyl,
• 3. (C₃-C₈) -cycloalkyl,
• 4. (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
• 5. (C₂-C₆) alkenyl,
• 6. (C₃-C₆) alkynyl,
• 7. R² and R³ together are (C₂-C₄) -alkyl,
• 8. a radical as defined under c) 7, which is substituted by halogen;
• d) R⁴
• 1. hydrogen,
• 2. (C₁-C₆) -alkyl,
• 3. (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
• 4. (C₃-C₁₀) alkenyl,
• e) R⁵
• 1. hydrogen,
• 2. a radical of the formula (II)
• f) R⁶ and R⁷ are the same or different
• 1. hydrogen,
• 2. (C₁-C₁₀) -alkyl,
• 3. (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
• 4. (C₁-C₁₀) -alkylamino,
• 5. (C₁-C₁₀) -alkylguanidino;
• g) R⁸
• 1. (C₆-C₁₀) -aryl,
• 2. (C₁-C₆) -alkyl,
• 3. (C₃-C₈) -cycloalkyl,
• 4. (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
• 5. (C₂-C₆) alkenyl,
• 6. a group defined under g) 1 and 4, which is substituted by one or more radicals from the series COR¹, halogen, nitro, cyano, (C₁-C₄) alkoxy or amino;
• h) R⁹
• 1. -NH- (CH₂) _o -NHR⁴
• 2. -O- (CH₂) _o -NH₂
• i) A represents a single or double bond;
• j) is BO, NR 3 or S;
• k) Y is a direct bond or an amino acid, preferably phenylalanine, 2-chlorophenylalanine, 3-chlorophenylalanine, 4-chlorophenylalanine, 2-fluorophenylalanine, 3-fluorophenylalanine, 4-fluorophenylalanine, tyrosine, O-methyltyrosine, β- (2-thienyl) alanine, glycine, cyclohexylalanine, leucine, isoleucine, valine, norleucine or phenylglycine, serine or cysteine;
• l) m is a number 1-5,
• m) n is a number 1-5,
• n) o is a number 1-10,
  and their physiologically acceptable salts.

2. Compounds of formula I according to claim 1, wherein

• a) R²
• 1. (C₁-C₆) -alkyl,
• 2. (C₂-C₆) alkenyl,
• 3. (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
• 4. (C₃-C₈) -cycloalkyl,
• 5. (C₃-C₆) -cycloalkyl- (C₁-C₃) -alkyl,
• 6. a defined under a) 3 and 5 radical which is substituted by one or more radicals from the series halogen, nitro, cyano, (C₁-C₄) alkoxy, COR¹ or amino; and
• c) R⁸
• 1. (C₆-C₁₀) -aryl,
• 2. (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
• 3. a c) under 1. and 2. defined radical which is substituted by one or more radicals from the series halogen, nitro, cyano, (C₁-C₄) alkoxy, amino or COR¹.

3. Compounds of formula I according to claims 1 or 2, wherein

• a) R¹-O- (C₁-C₁₀) -alkyl;
• b) R²
• 1. (C₆-C₁₀) -aryl-CH₂-,
• 2. (C₃-C₈) -cycloalkyl,
• 3. (C₃-C₈) -cycloalkyl-CH₂-;
• c) R³
• 1. (C₃-C₅) alkenyl,
• 2. phenyl- (C₁-C₃) -alkyl,
• 3. R² and R³ together are (C₂-C₃) alkyl;
• d) R⁴
• 1. hydrogen,
• e) R⁵
• 1. hydrogen,
• 2. a radical of formula (II) means
• f) R⁸
• 1. phenyl,
• 2. benzyl;
• g) R⁹
• 1. -NH- (CH₂) _o -NHR⁴,
• 2. -O- (CH₂) _o -NH _2, means;
• h) A is a single or double bond;
• i) Y
• 1. a single bond,
• 2. an amino acid selected from phenylalanine, β- (2-thienyl) alanine, O-methyltyrosine, glycine, cyclchexylalanine, leucine, isoleucine, valine, phenylglycine, serine or cysteine;
• j) o is a number from 1-10.

4. A process for the preparation of compounds of the formula I according to claims 1 to 3, characterized in that

• a) a compound of the formula (III) with a compound R³-X wherein
  R³ is hydrogen, (C₁-C₈) -alkyl, (C₃-C₈) -cycloalkyl, (C₆-C₁₀) -aryl (C₁-C₃) -alkyl, (C₂-C₆) -alkenyl or (C₃-C₆) -alkynyl means and
  X is a leaving group, such as halogen, mesylate or tosylate, with an organometallic base, such as n-butyllithium, s-butyl lithium, methyllithium or phenyllithium, sodium amide and alkali metal salts of organic nitrogen bases, such as lithium diisopropylamide, in an inert solvent, such as THF, ether, Toluene or dimethoxyethane, preferably at -78 ° C to give a compound of formula IV wherein R³ is as defined above;
• b) the compound of formula IV with an alcoholate of the formula R¹Na or R¹K, wherein
  R¹-OH, -O- (C₁-C₁₀) -alkyl, -O- (C₃-C₆) -alkenyl or -O- (C₆-C₁₀) -aryl (C₁-C₃) -alkyl,
  refluxing the corresponding alcohol to give a compound of formula V, wherein R¹ and R³ are as defined above;
• c₁) if in formula I R² and R³ together are (C₂-C₄) -alkyl, which may be optionally substituted by halogen, and in formula V R³ is (C₂-C₆) - alkenyl,
  the compound of formula V with J₂ or radical-generating reagents in methylene chloride or THF at room temperature to give a compound of formula VI wherein R is hydrogen or (C₁-C₃) alkyl and
  R¹ is as defined under b), reacted, and
• c₂) the resulting compound of formula VI with Raney nickel at 1atm H₂ and diisopropylethylamine as the base for 1 h at room temperature to give a compound of formula VII wherein R is hydrogen or (C₁-C₃) alkyl and
  R¹ is as defined under b) reacted;
• d) if in formula I R² and R³ together are not (C₂-C₄) -alkyl which may optionally be substituted by halogen,
  a compound of formula V wherein
  R¹-O- (C₁-C₁₀) -alkyl, -O- (C₃-C₆) -alkenyl or -O- (C₆-C₁₀) -aryl (C₁-C₃) -alkyl and
  R³ is hydrogen, (C₁-C₈) -alkyl, (C₃-C₈) -cycloalkyl, (C₆-C₁₀) -aryl (C₁-C₃) -alkyl, (C₂-C₆) -alkenyl or (C₃-C₆) -alkynyl means
  with a base, z. For example, sodium hydride, and then deprotonated with a compound R²-X, wherein
  R² is (C₁-C₁₀) -alkyl, (C₂-C₁₀) -alkenyl, (C₃-C₁₀) -alkynyl, (C₆-C₁₀) -aryl (C₁-C₃) -alkyl, (C₃-C₈) -cycloalkyl, ( C₄-C₁₀) -cycloalkyl- (C₁-C₄) -alkyl, (C₅-C₁₀) -cycloalkyl- (C₂-C₄) -alkenyl, (C₅-C₁₀) -cycloalkyl- (C₂-C₄) -alkynyl, - (CH₂ ) _m -B- (CH₂) _n -R⁵, (C₆-C₁₀) -aryl; (C₁-C₁₀) alkyl, (C₂-C₁₀) alkenyl, (C₃-C₁₀) alkynyl or - (CH₂) _m -B- (CH₂) _n -R⁵ monosubstituted with COR¹; (C₁-C₁₀) -alkyl, (C₂-C₁₀) -alkenyl, (C₃-C₁₀) -alkynyl or - (CH₂) _m -B- (CH₂) _n -R⁵, wherein 1 to all H atoms are substituted by halogen ; or (C₆-C₁₀) -aryl (C₁-C₃) -alkyl or (C₆-C₁₀) -aryl, which on aryl with 1 or 2 identical or different radicals selected from the group halogen, (C₁-C₄) alkoxy and nitro , Cyano are substituted,
  BO, NR 3 or S is,
  m is a number 1-5,
  n is a number 1-5,
  R⁵ is hydrogen or a radical of the formula II, and
  X is a suitable leaving group, e.g. Halogen, mesylate or tosylate,
  means
  converts to a compound of formula VIII, wherein R¹, R² and R³ are as defined under d);
• e) the compound of the formula VIII with a compound of the formula wherein
  Prot. For an amino protecting group, such. B. tert. Butoxycarbonyl, stands,
  R⁴ is hydrogen, (C₁-C₆) alkyl, (C₆-C₁₀) aryl (C₁-C₃) alkyl, (C₃-C₁₀) alkenyl, C (O) -O- (C₁-C₆) alkyl , -C (O) -O- (C₁-C₆) alkyl (C₆-C₁₀) aryl, or X is a suitable leaving group, e.g. As halogen, mesylate or tosylate, means
  with a base, z. As sodium hydride, deprotonated, and then with a compound of formula VII or VIII wherein
  R¹ is defined as under d),
  R² is (C₁-C₁₀) -alkyl, (C₂-C₁₀) -alkenyl, (C₃-C₁₀) -alkynyl, (C₆-C₁₀) -aryl (C₁-C₃) -alkyl, (C₃-C₈) -cycloalkyl, ( C₄-C₁₀) -cycloalkyl- (C₁-C₄) -alkyl, (C₅-C₁₀) -cycloalkyl- (C₂-C₄) -alkenyl, (C₅-C₁₀) -cycloalkyl- (C₂-C₄) -alkynyl, - (CH₂ ) _m -B- (CH₂) _n -R⁵, (C₆-C₁₀) -aryl; (C₁-C₁₀) alkyl, (C₂-C₁₀) alkenyl, (C₃-C₁₀) alkynyl or - (CH₂) _m -B- (CH₂) _n -R⁵ monosubstituted with COR¹; (C₁-C₁₀) -alkyl, (C₂-C₁₀) -alkenyl, (C₃-C₁₀) -alkynyl or - (CH₂) _m -B- (CH₂) _n -R⁵, wherein 1 to all H atoms are substituted by halogen ; or (C₆-C₁₀) -aryl (C₁-C₃) -alkyl or (C₆-C₁₀) -aryl, which on aryl with 1 or 2 identical or different radicals selected from the group halogen, (C₁-C₄) alkoxy and nitro , Cyano are substituted,
  BO, NR 3 or S is,
  m is a number 1-5,
  n is a number 1-5,
  R⁵ is hydrogen or a radical of the formula II, and
  R³ is hydrogen, (C₁-C₈) -alkyl, (C₃-C₈) -cycloalkyl, (C₆-C₁₀) -aryl (C₁-C₃) -alkyl or (C₃-C₆) -alkynyl,
  converts to a compound of formula IX, wherein R¹, R², R³, R⁴ and Prot are as defined under e) and
  R² and R³ together represent (C₂-C₄) alkyl optionally substituted with halogen;
• f) the amino protecting group Prot according to known methods, for. B. with tert. Butoxycarbonyl (Boc) under acidic conditions, preferably in trifluoroacetic acid (TFA) or with HCl in dimethoxyethane, or with TFA in dimethoxyethane or dichloromethane as a preferred solvent cleaved;
• g) optionally the resulting compound of the formula X having a free amino group wherein R¹, R², R³ and R⁴ are as defined under e) (formula IX), with a compound of formula II ' wherein
  R⁸ is (C₆-C₁₀) -aryl, (C₁-C₆) -alkyl, (C₃-C₈) -cycloalkyl, (C₆-C₁₀) -aryl (C₁-C₃) -alkyl, (C₂-C₆) -alkenyl; (C₆-C₁₀) -aryl or (C₆-C₁₀) -aryl (C₁-C₃) -alkyl having one or more radicals from the series COR¹, halogen, nitro, cyano, (C₁-C₄) - alkoxy or amino substituted, means
  R⁹ is -NH- (CH₂) _o -NHR⁴, -O- (CH₂) _o -NH₂ or means
  o is a number 1-10, and
  Y means a direct bond or an amino acid,
  Coupled according to the general methods of peptide chemistry, so as to obtain a compound of formula X ', wherein R¹, R², R³ and R⁴ are as defined under g) and R⁵ wherein R⁸, R⁹ and Y are as defined under g);
• h) optionally hydrogenating the double bond of the formulas X or X 'with sodium cyanoborohydride in lower alcohols, such as methanol, at room temperature and pH ~5 to give a compound of the formula XI, wherein R¹, R², R³, R⁴, R⁵ are as defined under g) and
  A means a single bond;
• i) optionally saponifying the compounds of the formulas X, X 'or XI under acidic or basic conditions or
  with HNR⁶R⁷, in which
  R⁶ and R⁷ are the same or different and are hydrogen, (C₁-C₁₀) alkyl, (C₆-C₁₀) - aryl- (C₁-C₃) alkyl, (C₁-C₁₀) -alkylamino or (C₁-C₁₀) -alkylguanidino .
  in high boiling solvents, e.g. As xylene or DMF, to obtain the compounds of formula I; and
• k) the compounds of formula I optionally converted by known methods into their physiologically acceptable salts.

5. Use of the compounds of the formula I according to claims 1 to 3 as a remedy. 6. Remedies containing a compound of formula I according to the claims 1 to 3.