HRP20000788A2 - Novel heterocyclically substituted amides with cysteine protease-inhibiting effect - Google Patents

Description

The proposed invention relates to new amides that are inhibitors of enzymes, especially cysteine proteases, such as calpain (= calcium-dependent cysteine proteases) and its isoenzymes, cathepsins, for example V and L.

Calpains are intracellular, proteolytic enzymes from the group of so-called cysteine proteases and are found in many cells. Calpains are activated by elevated calcium concentrations, with a difference between calpain I or μ-calpain, which is activated by μ-molar concentrations of calcium ions, and calpain II or m-calpain, which is activated by m-molar concentrations of calcium ions ( P. Johnson, Int. J. Biochem. 1990, 22(8), 811-22). It is now assumed that further calpain isoenzymes exist (K. Suzuki et al., Biol. Chem. Hoppe-Seyler, 1995, 376(9), 523-9).

Calpains are thought to play an important role in various physiological processes. These include cleavage of regulatory proteins, such as protein kinase C, cytoskeletal proteins, such as MAP 2 and spectrin, muscle proteins, protein degradation in rheumatoid arthritis, proteins in platelet activation, neuropeptide metabolism, proteins in mitosis, and others listed in M.J. Barrett et al., Life Sci. 1991, 48, 1659-69 and K. K. Wang et al., Trends in Pharmacol. Sci., 1994, 15, 412-9.

Elevated levels of calpain have been measured in various pathophysiological processes, for example: ischemia of the heart (e.g. cardiac infarction), renal or central nervous system (e.g. stroke), inflammation, muscular dystrophy, eye cataracts, central nervous system injury (e.g. trauma ), Alzheimer's disease, etc. (see K. K. Wang, above). It is assumed that there is a certain association of these diseases with elevated and persistent intracellular calcium levels. As a result, calcium-dependent processes are overactivated and are no longer subject to physiological regulation. Accordingly, overactivation of calpain can also cause pathophysiological processes.

Therefore, it was hypothesized that calpain enzyme inhibitors could be used to treat these diseases. This has been confirmed by various researches. Thus, Seung-Chyul Hong et al., Stroke 1994, 25(3), 663-9 and R. T. Bartus et al., Neurological Res. 1995, 17, 249-58, demonstrated the neuroprotective effect of calpain inhibitors in acute neurodegenerative disorders or ischemia, such as those occurring after stroke. Also, after experimental brain trauma, calpain inhibitors improve recovery from loss of memory and neuromotor disorders that occur (K. E. Saatman et al. Proc. Natl. Acad. Sci. USA, 1996, 93, 3428-3433). WITH. L. Edelstein et al., Proc. Natl. Acad. Sci. USA, 1995, 92, 7662-6, found a protective effect of calpain inhibitors on kidneys damaged by hypoxia. Yoshida, Ken Ischi et al., Jap. Circ. J. 1995, 59(1) 40-8, were able to demonstrate the beneficial effects of calpain inhibitors after cardiac damage caused by ischemia or reperfusion. Since calpain inhibitors inhibit the release of β-AP4 protein, its possible use as a therapeutic for Alzheimer's disease has been suggested (J. Higaki et al., Neuron, 1995, 14, 651-59). Interleukin 1α release was also prevented by calpain inhibitors (N. Watanabe et al., Cytokine 1994, 6(6), 597-601). In addition, calpain inhibitors have been found to exhibit cytotoxic effects on tumor cells (E. Shiba et al., 20th Meeting Int. Ass. Breast Cancer Res., Sendai Jp, 1994, Sept. 25-28, Int. J. Oncol. 5(Suppl.), 1994, 381).

Further possible uses of calpain inhibitors are listed in K. K. Wang, Trends in Pharmacol. Sci., 1994, 15, 412-8.

Calpain inhibitors have already been described in the literature. However, they are mostly irreversible or peptide inhibitors. As a rule, irreversible inhibitors are alkylating substances and have the disadvantage that they react indiscriminately in the body or are unstable. Therefore, these inhibitors often show unwanted side effects, such as toxicity, and accordingly they are limited in use or useless. Irreversible inhibitors include, for example, epoxides E 64 (E.B. McGowan et al., Biochem. Buiphys. Res. Commun. 1989, 158, 432-5), α-haloketones (H. Angliker et al., J. Med. Chem. 1992, 35, 216-20) or disulfides (R. Matsueda et al., Chem. Lett. 1990, 191-194).

Many known reversible inhibitors of cysteine proteases, such as calpain, are peptide aldehydes, especially dipeptide and tripeptide aldehydes, such as, for example, Z-Val-Phe-H (MDL 28170) (S. Mehdi, Trends in Biol. Sci. 1991, 16, 150-3). Under physiological conditions, peptide aldehydes have the disadvantage that they are often unstable due to high reactivity, can be rapidly metabolized, and are prone to nonspecific reactions that can cause toxic effects (J. A. Ferentz and B. Castro, Synthesis 1983, 676-78).

In JP 08183771 (CA 1996, 605307) and EP 520336, aldehydes derived from 4-piperidinoylamide and 1-carbonylpiperidino-4-ylamide are described as calpain inhibitors. However, the aldehydes claimed here, which are derived from the general structure I with heteroaromatically substituted amides, have never been described before.

Peptide ketone derivatives have also been described as inhibitors of cysteine proteases, especially calpain. Thus, for example, in the case of serine proteases ketone derivatives are known as inhibitors, where the keto group is activated with an electron-withdrawing group such as CF3. In the case of cysteine proteases, derivatives with keto groups activated by CF3 or similar groups are not very active or are inactive (M. R. Angelastro et al., J. Med. Chem. 1990, 33, 11-13). Surprisingly, in the case of calpain, only keto derivatives, in which, on the one hand, leaving groups in the α position cause irreversible inhibition, and on the other hand, the carboxylic acid derivative activates the keto group, have so far been found to be effective inhibitors (see M. R. Angelastro et al., see above; WO 92/11850; WO 92/12140; WO 94/00095 and WO 95/00535). However, of these keto amides and keto esters, only peptide derivatives have so far been described as effective (Zhaozhao Li et al., J. Med. Chem. 1993, 36, 3472-80; S. L. Harbenson et al., J. Med. Chem. 1994, 37, 2918-29 and see above M.R. Angelastro et al.).

Ketobenzamides are already known in the literature. Thus, the ketoester PhCO-Abu-COOCH2CH3 is described in WO 91/09801, WO 94/00095 and 92/11850. For the analogous phenyl derivative Ph-CONH-CH(CH2Ph)-CO-COCOOCH3, which is described in M. R. Angelastro et al., J. Med. Chem. 1990, 33, 11-13 , was found, however, to be only a weak calpain inhibitor. This derivative is also described in J.P. Burkhardt, Tetrahedron Lett., 1988, 3433-36. However, the significance of substituted benzamides has never been investigated.

In a number of therapies, such as in drip therapy, the active compounds are given intravenously as infusion solutions. For this purpose, it is necessary to have available substances, in this case calpain inhibitors, which have sufficient solubility in water so that an infusion solution can be prepared. However, many described calpain inhibitors have the disadvantage of showing little or no solubility in water and are therefore not suitable for intravenous administration. Active ingredients of this type can only be administered with the use of excipients designed to improve water solubility (see R.T. Bartus et al. J. Cereb. Blood Flow Metab. 1994, 14, 537-544). However, these excipients, for example polyethylene glycol, often cause side effects or are even intolerable. A non-peptide calpain inhibitor, which has satisfactory water solubility without excipients and therefore can probably be given with better tolerability, therefore has great advantages. Such inhibitors have not been described so far and would therefore be new.

In the proposed invention, substituted non-peptide aldehydes, ketocarboxylic acid esters and keto amide derivatives are described. These compounds are new and surprisingly show the possibility to obtain strong non-peptide inhibitors of cysteine proteases, such as for example calpain, by incorporation of rigid structural fragments. In addition, all proposed compounds of general formula I have at least one aliphatic amine radical and can therefore form salts with acids. A large number of these substances are soluble in water with the strength of a 0.5% solution at pH 0.4-5 and therefore show the profile required for intravenous administration, which is required, for example, for drip shock therapy.

The proposed invention relates to amides of general formula I

[image]

and their tautomeric and isomeric forms, possible enantiomeric and diastereomeric forms, and possible physiologically tolerable salts, in which the variables have the following meanings:

R1 can be hydrogen, C1-C6-alkyl, branched or unbranched, phenyl, naphthyl, quinolyl, pyridyl, pyrimidyl, pyrazyl, pyridazyl, quinazolyl, quinoxalyl, thienyl, benzo-thienyl, benzofuranyl, furanyl and indolyl, wherein the rings can also be substituted by 3 radicals R6 radicals, i

R2 is hydrogen, C1-C6-alkyl, branched or unbranched, O-C1-C6-alkyl, branched or unbranched, C2-C6-alkenyl, C2-C6-alkynyl, C1-C6-alkyl-phenyl, C2-C6- alkenyl-phenyl, C2-C6-alkynyl-phenyl, OH, Cl, F, Br, J, SF3, NO2, NH2, CN, COOH, COO-C1-C4-alkyl, NHCO-C1-C4-alkyl, NHCO- phenyl, CONHR9, NHSO2-C1-C4-alkyl, NHSO2-phenyl, SO2-C1-C4-alkyl and SO2-phenyl, and

R 3 can be NR 7 R 8 or a ring as

[image]

R 4 is -C 1 -C 6 -alkyl, branched or unbranched, which may also carry a phenyl, pyridyl, or naphthyl ring which is in turn substituted by at most two radicals R 6 , and

R5 is hydrogen, COOR11 and CO-Z wherein Z is NR12R13 and

[image]

and

R6 is hydrogen, C1-C4-alkyl, branched or unbranched, -O-C1-C4-alkyl, OH, Cl, F, Br, J, SF3, NO2, NH2, CN, COOH, COO-C1-C4-alkyl , -NHCO-C1-C4-alkyl, -NHCO-phenyl, -NHSO2-C1-C4-alkyl, -NHSO2-phenyl, -SO2- C1-C4-alkyl and –SO2-phenyl, and

R7 is hydrogen, C1-C6-alkyl, straight or branched, and which may be substituted with a phenyl ring which itself may also be substituted with one or two R10 radicals, and

R8 is hydrogen, C1-C6-alkyl, straight or branched, which may be substituted with a phenyl ring which itself may also be substituted with one or two R10 radicals, and

R9 is hydrogen, C1-C6-alkyl, straight or branched, which may also bear the substituent R16, or phenyl, pyridyl, pyrimidyl, pyridazyl, pyrazinyl, pyrazyl, naphthyl, quinolyl, imidazolyl, which may also bear one or two substituents R14 and

R10 can be hydrogen, C1-C4-alkyl, branched or unbranched, -O-C1-C4-alkyl, OH, Cl, F, Br, J, SF3, NO2, NH2, CN, COOH, COO-C1-C4- alkyl, -NHCO-C1-C4-alkyl, -NHCO-phenyl, -NHSO2-C1-C4-alkyl, -NHSO2-phenyl, -SO2-C1-C4-alkyl and –SO2-phenyl, and

R11 is hydrogen, C1-C6-alkyl, straight or branched, and which may be substituted with a phenyl ring which itself may also be substituted with one or two R10 radicals, and

R 12 is hydrogen, C 1 -C 6 -alkyl, straight or branched, and

[image]

R13 is hydrogen, C1-C6-alkyl, branched or unbranched, which may also be substituted with a phenyl ring which may also carry the radical R10, and

R14 is hydrogen, C1-C6-alkyl, branched or unbranched, -O-C1-C6-alkyl, branched or unbranched, OH, Cl, F, Br, J, SF3, NO2, NH2, CN, COOH, COO-C1 -C4-alkyl, or two radicals R14 can represent a bridge OC(R15)2O, i

R15 is hydrogen, C1-C6-alkyl, branched or unbranched, and

R16 can be a phenyl, pyridyl, pyrimidyl, pyridazyl, pyrazinyl, pyrazyl, pyrrolyl, naphthyl, quinolyl, imidazolyl ring, which can also carry one or two substituents R6, and

A is –(CH2)m-, –(CH2)m-O-(CH2)o-, –(CH2)o-S-(CH2)m-, –(CH2)o-SO-(CH2)m-, –(CH2 )o-SO2–(CH2)m-, -CH=CH-, -C≡C-, -CO-CH=CH-, -(CH2)o-SO–(CH2)m-, –(CH2)m -NNSO–(CH2)o-, –(CH2)o-CONH–(CH2)o-, –(CH2)m-NHSO2–(CH2)o-, -NH-CO-CH=CH-, –(CH2 )m-SO2NN–(CH2)o-, -CH=CH-CONH- and

[image]

R1-A together represent also

and

V is phenyl, pyridines, pyrimidines, pyrazines, imidazoles and thiazoles, and

x is 1, 2 or 3, and

n is the number 0, 1 or 2, i

m and o independently represent the number 0, 1, 2, 3 or 4.

The compounds of formula 1 can be used as racemates or as enantiomerically pure compounds, or as diastereomers. If enantiomerically pure compounds are desired, they can be obtained, for example, by classical resolution of racemates of compounds of formula I or their intermediates using a suitable optically active base or acid. On the other hand, enantiomeric compounds can also be produced using commercially available compounds, for example optically active amino acids such as phenylalanine, tryptophan and tyrosine.

The proposed invention also relates to compounds which are mesomeric or tautomeric with compounds of formula I, for example with those in which the keto group of formula I is present as an enol tautomer.

The proposed invention further relates to physiologically tolerable salts of compounds I, which can be obtained by reacting compound I with a suitable acid or base. Suitable acids and bases are described, for example, in “Fortschritte der Arzneimittelforschung, Volume 10, p. 224-285, Birkhäuser Verlag, Basel and Stuttgart, 1966. These include, for example, hydrochloric acid, citric acid, tartaric acid, lactic acid, phosphoric acid, methanesulfonic acid, acetic acid, formic acid, maleic acid, fumaric acid, etc. , or sodium hydroxide, lithium hydroxide, potassium hydroxide and tris.

Amides I according to the invention can be produced in various ways, which are shown in the synthesis scheme.

Synthesis scheme

Heterocyclic carboxylic acid II is connected to the corresponding amino alcohol III, which gives the corresponding amide IV. Conventional methods of peptide binding, described in C.R. Larock, Comprehensive Organic Transformations, VCH Publisher, 1989, p. 972 ff., or in Houben-Weyl, Methoden der organischen Chemie, 4th ed., E5, Ch. V. The reaction is carried out using "activated" derivatives of acid II, whereby the COOH group is converted into the COL group. L is a leaving group, such as Cl, imidazole or N-hydroxy-benzotriazole. This activated acid is then converted to amide IV using an amine. The reaction takes place in anhydrous, inert solvents, such as methylene chloride, tetrahydrofuran and dimethylformamide, at temperatures from -20 to +25oC.

These alcohol derivatives IV can be oxidized into new aldehyde derivatives I according to the invention. Various common oxidation reactions (see C. R. Larock, Comprehensive Organic Transformations, VCH Publisher, 1989, p. 604 et seq.) can be used for this purpose, such as, for example, Swern and Swern-analogous oxidations (T. T. Tidwell, Synthesis 1990, 857-70), sodium hypochloride/TEMPO (S. L. Harbenson et al., supra) or Dess-Martin (J. Org. Chem. 1983, 48, 4155). Advantageously, this reaction is carried out in inert aprotic solvents, such as dimethylformamide, tetrahydrofuran or methylene chloride, using oxidants such as DMSO/pyridine x SO3 or DMSO/oxalyl chloride at temperatures from -50 to +25°C, depending on the method (see literature above).

Alternatively, the carboxylic acid II can react with the aminohydroxamic acid derivative VI to give the benzamides VII. In this case, the same reaction procedure as for the preparation of compound IV is applied. Hydroxamic acid derivatives VI can also be obtained from protected amino acids V by reaction with hydroxylamine. In this process, the already described process for the preparation of amides can also be applied. Removal of the protecting group X, for example Boc, is carried out in a conventional manner, for example using trifluoroacetic acid. The thus obtained amide hydroxamic acid VII can be reduced to aldehydes I according to the invention. In this process, lithium aluminum hydride can be used as a reducing agent, for example, at a temperature of -60 to 0°C in inert solvents such as tetrahydrofuran or ether.

Analogously to the last procedure, carboxylic acids or acid derivatives can also be produced, such as esters IX (P = COOR', COSR'), which can also be converted by reduction to the aldehyde I according to the invention. These procedures are listed in R. C. Larock, Comprehensive Organic Transformations, VCH Publisher, 1989, p. 619-26.

Amides I according to the invention, which have heterocyclic substituents and carry a ketoamide or ketoester group, can be carried out in various ways, which are shown in synthesis schemes 2 and 3.

If appropriate, carboxylic acid esters IIa are converted to acids II using an acid or base such as lithium hydroxide, sodium hydroxide or potassium hydroxide in an aqueous medium or in a mixture of water and organic solvents such as alcohols or tetrahydrofuran at room temperature or at elevated temperature, such as 25-100°C.

Acids II are coupled to derivatives of α-amino acids, applying the usual conditions set forth, for example, in Houben-Weyle, Methoden der organischen Chemie, 4th ed., E5, Chap. V, and C.R. Larock, Comprehensive Organic Transformations, VCH Publisher, 1989, Chap. 9.

For example, the carboxylic acid II is converted to the "activated" acid derivative IIb =Y-COL, where L represents a leaving group, such as Cl, imidazole and N-hydroxy-benzotriazole, and then converted to the derivative XI by the addition of the amino acid derivative H2N- CH(R3)-COOR. This reaction is carried out in anhydrous, inert solvents such as methylene chloride, tetrahydrofuran and dimethylformamide at temperatures from -20 to +25oC.

Scheme 1

[image]

Derivatives XI, which are generally esters, are converted into ketocarboxylic acids XII analogously to the hydrolysis described above. Ketoesters I' are produced in a reaction analogous to the Dakin-West reaction, whereby the reaction is carried out according to the method of ZhaoZhao Li et al., J. Med. Chem. 1993, 36, 3472-80. In this process, the carboxylic acid as XII reacts with oxalic acid chloride monoester at elevated temperature (50-100oC) in solvents such as tetrahydrofuran and the product thus obtained is then reacted with a base such as sodium ethanolate in ethanol and at elevated temperature from 25-80oC, thereby obtaining ketoester I' in accordance with the invention. Ketoesters I' can be hydrolyzed as described above, for example to ketocarboxylic acids according to the invention.

The reaction to obtain ketobenzamide I' is also carried out analogously to the method described by ZhaoZhao Li et al. (see above). The keto group in I' is protected by the addition of 1,2-ethanedithiol with a Lewis acid catalyst, such as boron trifluoride etherate, in an inert solvent, such as methylene chloride, at room temperature to give a dithiane. These derivatives react with amines R3-H in polar solvents, such as alcohols, at temperatures of 0-80oC, giving ketoamides I (R4 is Z or NR7R8).

Scheme 2

[image]

An alternative method is shown in Scheme 2. Keto carboxylic acids II are reacted with amino-hydroxycarboxylic acid derivatives XIII (for the preparation of compounds XIII see S. L. Harbenson et al., J. Med. Chem. 1994, 37, 2918-29, or J. P. Burkhardt et al. al. Tetrahedron Lett. 1988, 29, 3433-3436) by the usual method for peptide coupling (see Houben-Weyl above), which gives amide XIV. These alcohol derivatives XIV can be oxidized into ketocarboxylic acid derivatives I according to the invention. For this purpose, various common oxidation reactions can be applied (see C. R, Larock, Comprehensive Organic Transformations, VCH Publisher, 1989, p. 604 et seq.) such as, for example, Swern and oxidations analogous to Swern, preferably dimethyl sulfoxide/ pyridine-sulfur trioxide complex in solvents such as methylene chloride or tetrahydrofuran, if appropriate with the addition of dimethyl sulfoxide, at room temperature or temperatures from -50 to 25°C (T.T. Tidwell, Synthesis 1990, 857-70) or sodium hypochloride/ TEMPO (S. L. Harbenson et al., see above).

If XIV are α-hydroxy esters (X = O-alkyl), they can be hydrolyzed to carboxylic acids XV, whereby the reaction is carried out analogously to the above methods, but preferably using lithium hydroxide in a mixture of water and tetrahydrofuran at room temperature. The production of other esters or amides XVI is carried out by reaction with amines under the coupling conditions already described. The alcohol derivative XVI can be oxidized, which again gives the ketocarboxylic acid derivatives I according to the invention.

The preparation of carboxylic acid ester II has already been described in some cases or is carried out according to conventional chemical methods.

Compounds in which X is a bond are produced by conventional aromatic coupling, for example Suzuki coupling with boric acid derivatives and halides with a palladium catalyst or coupling of aromatic halides with a copper catalyst. Alkyl bridge radicals (X = -(CH2)m-) can be produced by reduction of analogous ketones or by alkylation of an organolithium compound, e.g., ortho-phenyl-oxazolidine, or other organometallic compounds (see I. M. Dordor et al., J. Chem. Soc . Perkin Trans. I, 1984, 1247-52).

Derivatives with an ether bridge were produced by alkylation of the corresponding alcohols or phenols with halides.

Sulfoxides and sulfones can be obtained by oxidation of the corresponding thioethers.

Alkene and alkyne bridged compounds are produced, for example, by the Heck reaction from aromatic halides and the corresponding alkenes and alkynes (see I. Sakamoto et al., Chem. Pharm. Bull., 1986, 34, 2754-59).

Chalcones are formed by the condensation of acetophenone with aldehydes and can, if necessary, be converted into analogous alkyl derivatives by hydrogenation.

Amides and sulfonamides are produced from amines and acid derivatives analogously to the methods described above.

Dialkylaminoalkyl substituents are obtained by reductive amination of aldehyde derivatives with appropriate amines in the presence of boron hydrides such as BH3-pyridine-complex or NaBH3CN (A.F. Abdel-Magid, C.A. Maryanoff, K.G. Carson, Tetrahedron Lett. 10990, 31, 5595; A.E. Moormann, Synth. . Commun. 1993, 23, 789).

Amides I with heterocyclic substituents of the proposed invention are inhibitors of cysteine proteases, especially cysteine proteases such as calpains I and II and cathepsins B and L.

The inhibitory effect of amide I with heterocyclic substituents I was determined using usual enzyme experiments known from the literature, whereby the inhibitor concentration at which 50% of the enzyme activity was inhibited (= IC50) was determined as a measure of the effect. In this way, benzamides I were measured regarding the inhibition of calpain I, calpain II and cathepsin B.

Cathepsin B assay

Cathepsin B inhibition was determined analogously to the method described by S. Hasnain et al., J. Biol. Chem. 1993, 268, 235-40.

2 μl inhibitor solution, prepared from inhibitor and DMSO (final concentrations 100 μM to 0.01 μM) was added to 88 μl cathepsin B (cathepsin B from human liver, Calbiochem), diluted to 5 units in 500 μM buffer). This mixture is first incubated for 60 minutes at room temperature (25oC) and then the reaction is initiated by the addition of 10 μl of 10 mM Z-Arg-Arg-pNA (in a buffer with 10% DMSO). The reaction is observed for 30 minutes at 450 nM in a microtiter plate reader. IC50 values are then determined from the maximum gradients.

Examination of calpain I and II

Testing of the inhibitory properties of the calpain inhibitor was carried out in a buffer using 50 mM tris HCl, pH 7.5; 0.1 M NaCl; 1 mM dithiothreitol; 0.11 mM CaCl2; of the fluorogenic calpain substrate Suc-Leu-Tyr-AMC (25 mM dissolved in DMSO, Bachem/Switzerland). Human μ-calpain was isolated from erythrocytes and, after several steps of chromatography (DEAE-Sepharose, phenyl-Sepharose, Superdex 200 and Blue Sepharose), the >95% purity enzyme was examined by SDS-PAGE, Western blot analysis and N-terminal by sequencing. The fluorescence of the cleaved product 7-amino-4-methyl-coumarin (AMC) was observed in a Spex-Fluorolog fluorimeter at λ = 380 nm and λ = 460 nm. In the measurement range of 60 min, the substrate cleavage is linear and the autocatalytic activity of calpain is low if the experiments are performed at temperatures of 12°C. Inhibitors and calpain substrate are added to the experimental mixture as DMSO solutions, whereby the DMSO must not exceed a final concentration of 2%.

In the test mixture, 10 μl of substrate (250 μM final) and then 10 μl of μ-calpain (2 μg/ml final, ie 18 nM) are added to a 1 ml cuvette containing buffer. Calpain-mediated substrate cleavage is measured in 15–20 minutes. Then 10 μl of inhibitor (50-100 μM solution in DMSO) is added and cleavage inhibition is measured for 40 minutes.

Ki values were determined according to the following equation for reversible inhibition:

(Methods in Enzymology, )

Ki = I/(v0/vi) - 1; where I = concentration of inhibitor, v0 is the initial velocity before addition of inhibitor; vi is the reaction rate at equilibrium.

Velocity is calculated from v = release AMC/time, i.e. height/time.

Calpain is an intracellular cysteine protease. Calpain inhibitors must cross the cell membrane to prevent degradation of intracellular proteins by calpain. Some known calpain inhibitors, such as E 64 and leupeptin, hardly cross cell membranes, and although they are good calpain inhibitors, they show modest activity in cells. The goal is to find compounds that pass through membranes better. As evidence that calpain inhibitors pass through the membrane, human platelets were used.

Degradation of tyrosine kinase pp60src in platelets by calpain

After platelet activation, the tyrosine kinase pp60src is cleaved with calpain. This trial was investigated in detail by Oda et al. in J. Biol. Chem., 1993, Vol 268, 12603-12608. In doing so, it was shown that the cleavage of pp60src can be effectively prevented with calpeptin, a calpain inhibitor. The effectiveness of our substances in the cell has been tested according to this publication. Fresh blood mixed with citrate is centrifuged for 15 min at 200 g. Platelet-rich plasma is collected and diluted 1:1 with platelet buffer (platelet buffer: 68 mM NaCl, 2.7 mM KCl, 0.5 mM MgCl2 x 6 H2O, 0.24 mM NaH2PO4 x N2O, 12 mM NaNSO3, 5.6 mM glucose, 1 mM EDTA, pH 7.4). After centrifugation and washing with platelet buffer, the platelets are adjusted to 107 cells/ml. Isolation of human platelets was performed at room temperature.

In the test mixture, isolated platelets (2 x 106) were pre-incubated for 5 minutes at 37°C with different concentrations of inhibitor (dissolved in DMSO). Then platelets were activated with 1 μM ionophore A23187 and 5 mM CaCl2. After 5 min of incubation, the platelets were briefly centrifuged at 13000 rpm and the pellet was taken up in SDS sample buffer (SDS sample buffer: 20 mM Tris-HCl, 5 mM EDTA, 5 mM EGTA, 1 mM DTT, 0.5 mM PMSF, 5 μg/ml leupeptin, 10 μg/ml pepstatin, 10% glycerol and 1% SDS). Proteins were separated on a 12% gel and pp60src and its 52 kDa and 47 kDa cleavage products were identified by Western blotting. The polyclonal rabbit anti-Cis-src (rr600-src) antibody used was from Biomol Feinchemikalien (Hamburg). This primary antibody was detected using HRP-linked goat second antibody (Boehringer Mannheim, FRG). Western blot testing was performed in a known manner.

Quantification of pp60src cleavage was performed by densitometry, and unactivated platelets (control 1: no cleavage) and platelets mixed with ionophore and calcium (control 2: response to 100% cleavage) were used as controls. The ED50 values correspond to the inhibitor concentration at which the intensity of the colored reaction is reduced by 50%.

Cell death in cortical neurons induced by glutamate

This study was performed as described by Choi D.W., Maulucci-Gedde M.A., and Kriegstein A.R., "Glutamate neurotoxicity in cortical cell culture". J. Neurosci. 1989, 7, 357-368.

Halves of the cortex were extracted from 15-day-old mouse embryos and individual cells were obtained enzymatically (trypsin). These cells (glia and cortical neurons) were inoculated in 24-well plates. After three days (laminin-coated plates) or seven days (ornithine-coated plates), mitosis processing was performed using FDU (5-fluoro-2-deoxyuridine). 15 days after cell preparation, cell death was induced by the addition of glutamate (15 minutes). After glutamate removal, calpain inhibitors are added. 24 hours later, cell damage was determined by determining lactate dehydrogenase (LDH) in the cell culture supernatant.

It is assumed that calpain also plays a role in apoptotic cell death (M. K. T. Squier et al. J. Cell. Physiol. 1994, 159, 229-237; T. Patel et al. Faseb Journal 1996, 590, 587-597). Therefore, in the following model, cell death was induced with calcium in the presence of a calcium ionophore in a human cell line. Calpain inhibitors must enter the cell and inhibit calpain there to prevent induced cell death.

Calcium-mediated cell death in NT2 cells

Cell death can be induced in the human cell line NT2 (Stratagene GmbH) by calcium in the presence of ionophore A 23187. 105 cells/well are placed in microtiter plates 20 hours before the experiment. After this period, the cells are incubated with various concentrations of inhibitors in the presence of 2.5 µM ionophore and 5 mM calcium. After 5 hours, 0.05 ml of XTT (set II for cell proliferation, Boehringer Mannheim) was added to the reaction mixture. The optical density is determined approximately 17 hours later according to the manufacturer's instructions, in an Easy Reader EAR 400 by SLT. The optical density at which half the cells died was calculated from two controls with cells without inhibitor, which were incubated in the absence and in the presence of the ionophore.

In numerous neurological diseases or psychological disorders, there is an increased activity of glutamate, which leads to a state of excessive stimulation or toxic effects in the central nervous system (CNS). Glutamate mediates its effects using various receptors. Two of these receptors are classified according to specific agonists as the MMDA receptor and the AMPA receptor. Antagonists against these glutamate-mediated effects can therefore be used to treat these diseases, especially for therapeutic application against neurodegenerative diseases such as Huntington's disease and Parkinson's disease, neurotoxic disorders after hypoxia, anoxia, ischemia and after lesions, such as those arising after shock and trauma, or alternatively as antiepileptics (see Arzneim. Forschung 1990, 40, 511-514; TIPS, 1990, 11, 334-338; Drugs of Future 1989, 14. 1059-1071).

Protection against cerebral overstimulation with excitatory amino acids (NMDA or AMPA antagonism in mice)

Intracerebral application of excitatory amino acids (EAA) induces massive overstimulation so that in a short time it leads to spasms and death of animals (mice). These symptoms can be inhibited by systemic, eg intraperitoneal, administration of centrally active compounds (EAA antagonists). Since excessive activation of EAA receptors of the central nervous system plays an important role in the pathogenesis of various neurological disorders, the demonstrated EAA antagonism in vivo can be used to conclude on the possible therapeutic use of substances against CNS disorders of this type. As a measure of the efficacy of the substance, the ED50 value at which 50% of the animals were symptom-free as a result of the previous i.p. applications of fixed doses of NMDA or AMPA test substance.

Heterocyclic substituted amides I are inhibitors of cysteine derivatives such as calpain I or II and cathepsin B or L and can therefore be used to suppress diseases associated with increased enzymatic activity of calpain enzymes or cathepsin enzymes. The proposed amides I can, accordingly, be used for the treatment of neurodegenerative processes that occur after ischemia, trauma, subarachnoid hemorrhages and strokes, and neurodegenerative diseases such as multiple infarct dementia, Alzheimer's disease, Huntington's disease and epilepsy, and in addition for the treatment of heart injury after cardiac ischemia, injury and reperfusion after vascular occlusion, kidney injury after renal ischemia, muscular dystrophy, injury occurring due to proliferation of smooth muscle cells, coronary vasospasms, cerebral vasospasms, eye cataracts, restenosis of blood streams after angioplasty . In addition, amides I can be used in the chemotherapy of tumors and their metastases and for the treatment of diseases in which there is an elevated level of interleukin 1, such as inflammation and rheumatic disorders.

In addition to the usual pharmaceutical excipients, the pharmaceutical preparations according to the invention contain a therapeutically effective amount of compounds I.

For topical external application, for example in dusting powders, ointments or sprays, the active compounds can be contained in the usual concentrations. As a rule, active compounds are contained in an amount of 0.001 to 1 mass. %, preferably 0.001 to 0.1 wt. %.

In the case of internal application, the preparations are given in single doses. In a single dose, 0.1 to 100 mg per kg of body weight is prescribed. The preparation can be given in one or more daily doses, depending on the nature and severity of the disorder.

According to the desired method of application, the pharmaceutical preparations according to the invention contain, in addition to the active compound, the usual excipients and diluents. For local external application, pharmaceutical excipients such as ethanol, isopropanol, ethoxylated castor oil, ethoxylated hydrogenated castor oil, polyacrylic acid, polyethylene glycol, polyethylene glycol stearate, ethoxylated fatty alcohols, paraffin oil, petrolatum and wool grease can be used. Suitable for internal application are, for example, lactose, propylene glycol, ethanol, starch, talc and polyvinylpyrrolidone.

Antioxidants such as tocopherol and butylated hydroxyanisole as well as butylated hydroxytoluene, flavor enhancers, stabilizers, emulsifiers and lubricants can be added.

Substances contained in the preparation, in addition to the active compound and substances used for the production of pharmaceutical preparations, are toxicologically acceptable and compatible with the relevant active compound. Pharmaceutical preparations are produced in a conventional manner, for example by mixing the active compound with conventional excipients and diluents.

The pharmaceutical compositions can be administered in a variety of ways, for example, orally, parenterally such as intravenously or by infusion, subcutaneously, intraperitoneally, and topically. Preparations in the form of tablets, emulsions, infusion and injection solutions, pastes, ointments, gels, creams, lotions, powders and sprays are thus possible.

EXAMPLES

Example 1

2-((4-phenylpiperazin-1-yl)methyl)benzoic acid N-(3-phenyl-propan-1-al-2-yl)amide

a) Methyl 2-(4-phenyl-1-piperazinylmethyl)benzoate

10.0 g of methyl 2-chloromethylbenzoate, 15 g of potassium carbonate, 8.8 g of N-phenylpiperazine and 18-crown-6 at the tip of a knife in 200 ml of DMF are heated for 5 h at 100°C and then stirred for 60 h at room temperature. Excess potassium carbonate is filtered off, the filtrate is concentrated, and the residue is partitioned between water and ethyl acetate. By drying the organic phase over magnesium sulfate and removing the solvent, 16.8 g (100%) of the product was obtained.

b) 2-(4-phenyl-1-piperazinylmethyl)benzoic acid

16.8 g of intermediate la was placed in 150 ml of THF, and 1.7 g of LiOH in 150 ml of water was added at room temperature. The cloudy solution was clarified by adding 10 ml of MeOH. The reaction mixture was stirred for 12 h at room temperature and hydrolyzed with an equimolar amount of 1 M HCl. The reaction mixture was evaporated to dryness and the residue was taken up in methanol/toluene. Removal of the solvent gave 15.2 g (86%) of the product, which still contained salt.

c) 2-((4-phenylpiperazin-1-yl)methyl)benzoic acid N-(3-phenylpropan-1-ol-2-yl)amide

3.0 g of intermediate 1b and 3 ml of triethylamine were placed in 50 ml of DMF. Add 5 g of sodium sulfate and mix the mixture for 30 minutes. At 0°C, 1.5 g of phenylalaninol, 1.4 g of HOBT and 2.1 g of EDC were added sequentially and the mixture was stirred overnight at room temperature. The reaction mixture is poured into distilled water, made alkaline with NaNSO3, saturated with NaCl and extracted three times with 100 ml of methylene chloride. The organic phases are washed twice with water and dried over magnesium sulfate. Removal of the solvent gives 2.5 g (59%) of the product.

d) 2-((4-phenylpiperazin-1-yl)methyl)benzoic acid N-(3-phenylpropan-1-al-2-yl)amide

2.3 g of intermediate 1c was placed in 50 ml of DMSO in the presence of 2.4 g of triethylamine, and 2.5 g of SO2/pyridine complex was added. The mixture is stirred overnight at room temperature. The mixture is poured into 250 ml of distilled water, made alkaline with NaHCO3, saturated with NaCl and extracted with 100 ml of methylene chloride, and the organic phase is dried over magnesium sulfate. The residue after removal of the solvent was dissolved in THF and the hydrochloride was precipitated with HCl in dioxane. The precipitate is filtered off with suction and washed several times with ether, yielding 1.9 g (71%) of the product.

1H-NMR (d6-DMSO): δ = 2.9 (2H), 3.0-3.3 (8H), 4.1-4.5 (2H), 4.7 (1N), 6.8 -7.7 (14N), 9.3 (1N), 9.8 (1N) ppm.

Example 2

2-((4-benzylpiperazin-1-yl)methyl)benzoic acid N-(3-phenylpropan-1-al-2-yl)amide

a) Methyl 2-((4-benzyl-1-piperazinyl)methyl)benzoate

10.0 g of methyl 2-chlorobenzoate and 9.6 g of N-benzylpiperazine are reacted in 200 ml of DMF in the presence of 15 g of potassium carbonate at 100°C analogously to example 1a, which gives 17.6 g (100%) of the product.

b) 2-((4-benzyl-1-piperazinyl)methyl)benzoic acid

17.5 g of intermediate 2a in 150 ml of THF is hydrolyzed with 1.6 g of LiOH in 150 ml of water analogously to example 1b, which gives 9.1 g (54%) of the product.

c) 2-((4-benzylpiperazin-1-yl)methyl)benzoic acid N-(3-phenylpropan-1-ol-2-yl)amide

3.0 g of intermediate 2b is reacted in 60 ml of DMF with 3 ml of triethylamine, 1.5 g of phenylalaninol, 1.3 g of HOBT and 2.0 g of EDC analogously to example 1c, which gives 2.0 g (46%) of products.

d) 2-((4-benzylpiperazin-1-yl)methyl)benzoic acid N-(3-phenylpropan-1-al-2-yl)amide

1.5 g of intermediate 2c is oxidized in 40 ml of DMSO with 1.9 g of SO3/pyridine complex in 20 ml of DMSO in the presence of 2.3 ml of triethylamine analogously to example 1d, which gives 0.4 g (21%) of the product in the form fumarate.

1H-NMR (d6-DMSO): δ = 2.1-2.3 (8H), 2.9-3.0 (1N), 3.3-3.6 (6H), 4.5 (1N) , 6.6 (2H), 7.1-7.7 (14N), 9.7 (1N), 10.3 (1N) ppm.

Example 3

2-((4-benzylpiperazin-1-yl)methyl)benzoic acid H-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)amide

a) 2-((4-benzylpiperazin-1-yl)methyl)benzoic acid N-(1-carbamoyl-1-ol-3-phenylpropan-2-yl)amide

1.5 g of intermediate 2b is reacted in 40 ml of DMF with 0.7 ml of triethylamine, 1.0 g of 3-amino-2-hydroxy-4-phenylbutyramide hydrochloride, 0.6 g of HOBT and 0.9 g of EDC analogously to example 1c, which gives 0.8 g (38%) of the product.

b) 2-((4-benzylpiperazin-1-yl)methyl)benzoic acid N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)amide

0.7 g of intermediate 3a is oxidized in 20 ml of DMSO with 0.7 g of SO3/pyridine complex in the presence of 0.8 g of triethylamine analogously to example 1d, which gives 0.1 g (18%) of the product in the form of the free base.

1H-NMR (d6-DMSO): δ = 2.3 (4H), 2.8-3.5 (8H), 5.3 (1N), 6.7-7.5 (16H), 7.8 (1N), 8.1 (1N), 10.3 (1N) ppm.

Example 4

2-(4-((3-methylphenyl)piperazin-1-yl)methyl)benzoic acid N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)amide

a) Methyl 2-(4-((3-methylphenyl)-1-piperazinyl)-methyl)benzoate

4.0 g of methyl 2-chloromethylbenzoate and 4.4 g of 3-methyl-phenyl-piperazine are heated for 3 h in 200 ml of DMF in the presence of 4.5 g of potassium carbonate at 140°C. The reaction mixture was poured into water and extracted three times with ethyl acetate. The combined organic phases are washed three times with saturated NaCl solution, dried over magnesium sulfate and concentrated to give 6.5 g (92%) of the product.

b) 2-(4-((3-methylphenyl)-1-piperazinyl)methyl)-benzoic acid

5.9 g of intermediate 4a is dissolved in 75 ml of THF and hydrolyzed with 0.9 g of LiOH in 75 ml of water analogously to example 1b, which gives 2.9 g (51%) of the product.

c) 2-(4-((3-methylphenyl)piperazin-1-yl)methyl)-benzoic acid N-(1-carbamoyl-1-ol-3-phenylpropan-2-yl)amide

1.8 g of intermediate 4b was placed in 50 ml of DMF in the presence of 2.7 ml of triethylamine, and 0.8 g of HOBT. 1.3 g of 3-amino-2-hydroxy-4-phenylbutyramide hydrochloride and 1.2 g of EDC were successively added analogously to example 1c, resulting in 1.4 g (50%) of the product.

d) 2-(4-((3-methylphenyl)piperazin-1-yl)methyl)-benzoic acid N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)amide

1.2 g of intermediate 4c was dissolved in 30 ml of DMSO and oxidized with 1.6 g of SO3/pyridine complex in the presence of 1.5 ml of triethylamine analogously to example 1d, which gave 1.0 g (83%) of the product.

MS: m/e = 484 (M+)

Examples 5 and 6 were synthesized analogously to example 1.

Example 5

3-((4-phenylpiperazin-1-yl)methyl)benzoic acid N-(3-phenylpropan-1-al-2-yl)amide fumarate

1H-NMR (d6-DMSO): δ = 2.5 (4H), 2.9 (1H), 3.2 (4H), 3.3 (1N), 3.7 (2N), 4.5 ( 1H), 6.6 (2H), 6.75 (1N), 6.9 (2H), 7.2 (2N), 7.2-7.3 (5N), 7.45 (1N), 7 .55 (1N), 7.75 (1N), 7.8 (2N), 8.9 (1N), 9.7 (1N) ppm.

Example 6

3-((4-(2-tert-butyl-4-trifluoromethylpyrimidin-6-yl)-homopiperazin-1-yl)methyl)benzoic acid N-(3-phenyl-

propan-1-al-2-yl) amide

MS: m/e = 568 (M++l)

Example 7

4-(N-(3,4-dioxomethylene)benzyl-N-methylaminomethyl)-benzoic acid N-(3-phenylpropan-1-al-2-yl)amide

a) 4-(N-(3,4-dioxomethylene)benzyl-N-methylaminomethyl)-benzoic acid

11.5 g of N-(3,4-dioxomethylene)benzyl-N-methylamine and 15.5 g of triethylamine were placed in, and 15.0 g of 4-bromomethyl-benzoic acid in 100 ml of THF was added. The reaction mixture is briefly heated to reflux and then stirred for 15 h at room temperature. When the salt is filtered off, the mother liquor is concentrated and the residue is dissolved in ethyl acetate and washed with water. The aqueous phase was basified and extracted several times with ethyl acetate to give 6.6 g (32%) of the product as a white solid.

b) 4-(N-(3,4-dioxomethylene)benzyl-N-methylaminomethyl)-benzoic acid N-(3-phenylpropan-1-ol-2-yl)amide

4.4 g of intermediate 5a was placed in 50 ml of DMF in the presence of 2.9 g of triethylamine and 1.8 g of HOBT, 2.0 g of phenylalaninol and 2.8 g of EDC were added in sequence analogously to example 1c, thereby 2.3 g (40%) of the product is obtained.

c) 4-(N-(3,4-dioxomethylene)benzyl-N-methylamino-methyl)-benzoic acid N-(3-phenylpropan-1-al-2-yl)-amide

2.0 g of intermediate 5b was dissolved in 60 ml of DMSO and oxidized with 2.1 g of SO3/pyridine complex in the presence of 1.8 ml of triethylamine analogously to example 1d, which gave 1.3 g (68%) of the product.

1H-NMR (CF3COOD): δ = 2.9 (3H), 3.2 (2H), 4.3-4.9 (5H), 6.1 (2H), 6.6 (1N), 6, 9 (3H), 7.2-7.4 (5N), 7.8 (2H), 8.25 (2H) ppm.

MS: m/e = 430 (M+)

Examples 8-28 were produced analogously to Example 7.

Example 8

4-(M-benzyl-N-methylaminomethyl)benzoic acid N-(3-phenylpropan-1-al-2-yl)amide

1H-NMR (CF3COOD): δ = 2.9 (ZN), 3.2 (2N), 4.3-5.0 (5N), 6.7 (1N), 7.25-7.5 (8H ), 7.55 (2N), 7.8 (2N), 8.2 (2N) ppm.

MS: m/e = 386 (M+)

Example 9

4-(N-(4-methoxy)benzyl-N-methylaminomethyl)benzoic acid

N-(3-phenylpropan-1-al-2-yl)amide

1H-NMR (CF3COOD): δ = 2.9 (3N), 3.3 (2N), 4.0 (ZN), 4.3-4.9 (5N), 6.7 (1N), 7, 1-7.4 (7N), 7.5 (2N), 7.8 (2N), 8.2 (2N) ppm.

MS: m/e = 416 (M+)

Example 10

4-(N-benzyl-N-methylaminomethyl)benzoic acid N-(3-butan-1-al-2-yl)amide

1H-NMR (CF3COOD): δ = 1.1 (3H), 1.6 (2N), 2.0 (2N), 2.9 (ZN), 4.3-4.5 (ZN), 4, 7 (1N), 4.8 (1N), 6.6 (1N), 7.3-7.6 (5H), 7.8 (2N), 8.3 (2N) ppm.

MS: m/e = 338 (M+)

Example 11

4-(N-(3,4-dioxomethylene)benzyl-N-methylaminomethyl)-benzoic acid N-(3-butan-1-al-2-yl)amide

1H-NMR (CF3COOD): δ = 1.1 (3N), 1.6 (2N), 1.9 (2N), 2.9 (3N), 4.25-4.6 (4N), 4, 75 (1N), 6.1 (2H), 6.6 (1N), 6.9 (3N), 7.8 (2N), 8.3 (2N) ppm.

MS: m/e = 382 (M+)

Example 12

4-(N-(4-methoxy)benzyl-N-methylaminomethyl)benzoic acid N-(3-butan-1-al-2-yl)amide

MS: m/e = 368 (M+)

Example 13

4-(N-(3,4-dioxomethylene)benzyl-N-methylaminomethyl)-benzoic acid N-(3-cyclohexylpropan-1-al-2-yl)amide

1H-NMR (CF3COOD): δ = 1.0-2.0 (13H), 2.9 (3N), 4.3-4.9 (4N), 6.1 (2N), 6.6 (1N ), 6.9 (3N), 7.8 (2N), 8.3 (2N)

ppm.

MS: m/e = 436 (M+)

Example 14

4-(N-(4-benzyl-M-methylaminomethyl)benzoic acid N-(3-cyclohexylpropan-1-al-2-yl)amide

1H-NMR (d6-DMSO): δ = 1.0-1.8 (13H), 2.1 (3N), 3.4 (2N), 3.5 (2N), 4.3 (1N), 7.1-7.4 (5N), 7.5 (2N), 7.8 (2N), 8.8 (1N), 9.5 (1N) ppm.

Example 15

4-(N-(4-methoxy)benzyl-M-methylaminomethyl)benzoic acid N-(3-cyclohexylpropan-1-al-2-yl)amide

1H-NMR (CDCl3): δ = 1.0-1.8 (13H), 2.1 (3N), 3.4 (2N), 3.5 (2N), 3.7 (3N), 4, 3 (1N), 6.8 (2H), 7.25 (2N), 7.5 (2N), 7.9 (2N), 8.8 (1N), 9.5 (1N) ppm.

Example 16

4-((2-phenylpyrrolid-1-yl)methyl)benzoic acid N-(3-cyclohexylpropan-1-al-2-yl)amide

MS: m/e = 420 (M+)

Example 17

4-((2-phenylpyrrolid-1-yl)methyl)benzoic acid N-(3-butan-1-al-2-yl)amide

MS: m/e = 364 (M+)

Example 18

4-((2-phenylpyrrolid-1-yl)methyl)benzoic acid N-(3-phenylpropan-1-al-2-yl)amide

MS: m/e = 412 (M+)

Example 19

4-((1,2,3,4-dihydroquinolin-1-yl)methyl)benzoic acid N-(3-cyclohexylpropan-1-al-2-yl)amide

1H-NMR (CDCl3): δ = 1.0-1.9 (13H), 2.0 (2N), 2.8 (2N), 3.3 (2N), 4.5 (2N), 4, 8 (1N), 6.4 (1N), 6.5 (2N), 7.0 (2N), 7.4 (2N), 7.8 (2N), 9.7 (1N) ppm.

MS: m/e = 404 (M+)

Example 20

4-((1,2,3,4-dihydroquinolin-1-yl)methyl)benzoic acid N-(3-phenylpropan-1-al-2-yl)amide

1H-NMR (d6-DMSO): δ = 1.9 (2H), 2.75 (2N), 2.9 (1H), 3.3 (1H), 3.4 (2H), 4.4 ( 1N), 4.5 (2H), 6.3 (2N), 6.8 (2N), 7.1-7.25 (5N), 7.3 (2N), 7.7 (2N), 8 .8 (1N), 9.5 (1H) ppm.

MS: m/e = 398 (M+)

Example 21

4-((1,2,3,4-dihydroquinolin-1-yl)methyl)benzoic acid N-(3-butan-1-al-2-yl)amide

1H-NMR (d6-DMSO): δ = 0.9 (3H), 1.2-2.0 (6H), 2.7 (2N), 3.3 (2N), 4.2 (1H), 4.5 (2N), 6.4 (2N), 6.8 (2N), 7.3 (2N), 7.8 (2N), 8.8 (1H), 9.5 (1H) ppm.

MS: m/e s= 350 (M+)

Example 22

4-((1,2,3,4-dihydroisoquinolin-2-yl)methyl)benzoic acid N-(3-cyclohexylpropan-1-al-2-yl)amide

1H-NMR (d6-DMSO): δ = 0.9-1.8 (13H), 2.7-2.9 (4H), 3.6 (2N), 3.75 (2N), 4.4 (1N), 6.9-7.1 (4H), 7.4 (2N), 7.8 (2N), 8.8 (1H), 9.5 (1H) ppm.

MS: m/e = 404 (M+)

Example 23

4-((1,2,3,4-dihydroisoquinolin-2-yl)methyl)benzoic acid N-(3-phenylpropan-1-al-2-yl)amide

1H-NMR (d6-DMSO): δ = 2.7 (2H), 2.8 (2H), 2.9 (1N), 3.2 (1H), 3.5 (2H), 3.7 ( 2H), 4.5 (1H), 6.9-7.1 (4H), 7.2-7.3 (5H), 7.5 (2H), 7.75 (2H), 8.8 ( 1H), 9.5 (1N) ppm.

MS: m/e = 398 (M+)

Example 24

4-((1,2,3,4-dihydroisoquinolin-2-yl)methyl)benzoic acid M-(3-butan-1-al-2-yl)amide hydrochloride

1H-NMR (d6-DMSO): δ = 0.9 (3H), 1.2-2.0 (4H), 3.0 (1N), 3.3 (2H), 3.6 (1N), 4.1-4.6 (5H), 7.2 (4H), 7.8 (2H), 8.0 (2H), 9.0 (1N), 9.5 (1N), 11.75 ( 1N) ppm.

Example 25

4-((6,7-dimethoxy-1,2,3,4-dihydroisoquinolin-2-yl)-methyl)-benzoic acid N-(3-cyclohexylpropan-1-al-2-yl)amide

1H-NMR (d6-DMSO): δ = 0.9-1.9 (13H), 2.7 (4H), 3.4 (2H), 3.6 (3H), 3.65 (2H), 3.7 (3H), 4.3 (1N), 6.5 (1N), 6.6 (1N), 7.5 (2H), 7.8 (2H), 8.8 (1H), 9 .5 (1N) ppm.

MS: m/e = 464 (M+)

Example 26

4-((6,7-dimethoxy-1,2,3,4-dihydroisoquinolin-2-yl)-methyl)-benzoic acid N-(3-phenylpropan-1-al-2-yl)amide

1H-NMR (d6-DMSO): δ = 2.7 (4H), 2.9 (1N), 3.25 (1N), 3.6 (6H), 3.7 (2H), 4.5 ( 1N), 6.6 (1N), 6.7 (1N), 7.2-7.3 (5H), 7.4 (2H), 7.8 (2H), 8.9 (1N), 9 .6 (1N) ppm.

MS: m/e = 458 (M+)

Example 27

4-((6,7-dimethoxy-1,2,3,4-dihydroisoquinolin-2-yl)-methyl)-benzoic acid N-(3-butan-1-al-2-yl)amide

1H-NMR (d6-DMSO): δ = 0.9 (3N), 1.4 (2N), 1.5-1.8 (2N), 2.7 (4N), 3.4 (2N), 3.7 (3N), 3.75 (3N), 3.8 (2N), 4.3 (1N), 6.6 (1H), 6.7 (1H), 7.4 (2H), 7 .8 (2N), 8.8 (1N), 9.5 (1H) ppm.

MS: m/e = 410 (M+)

Example 28

2-((1,2,3,4-dihydroquinolin-1-yl)methyl)benzoic acid N-(3-butan-1-al-2-yl)amide

MS: m/e = 441 (M+)

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Claims (23)

1. Amide of the general formula I [image] and its tautomeric and isomeric forms, possible enantiomeric and diastereomeric forms, and possible physiologically tolerable salts, indicated that therein the variables have the following meanings: R1 can be hydrogen, C1-C6-alkyl, branched or unbranched, phenyl, naphthyl, quinolyl, pyridyl, pyrimidyl, pyrazyl, pyridazyl, quinazolyl, quinoxalyl, thienyl, benzo-thienyl, benzofuranyl, furanyl and indolyl, wherein the rings can also be substituted by 3 radicals R6 radicals, i R2 is hydrogen, C1-C6-alkyl, branched or unbranched, O-C1-C6-alkyl, branched or unbranched, C2-C6-alkenyl, C2-C6-alkynyl, C1-C6-alkyl-phenyl, C2-C6- alkenyl-phenyl, C2-C6-alkynyl-phenyl, OH, Cl, F, Br, J, SF3, NO2, NH2, CN, COOH, COO-C1-C4-alkyl, NHCO-C1-C4-alkyl, NHCO- phenyl, CONHR9, NHSO2-C1-C4-alkyl, NHSO2-phenyl, SO2-C1-C4-alkyl and SO2-phenyl, and R 3 can be NR 7 R 8 or a ring as [image] R4 is -C1-C6-alkyl, branched or unbranched, which may also carry a phenyl, pyridyl, thienyl, cyclohexyl, indolyl or naphthyl ring which is in turn substituted by at most two R6 radicals, and R5 is hydrogen, COOR11 and CO-Z wherein Z is NR12R13 and [image] and R6 is hydrogen, C1-C4-alkyl, branched or unbranched, -O-C1-C4-alkyl, OH, Cl, F, Br, J, SF3, NO2, NH2, CN, COOH, COO-C1-C4-alkyl , -NHCO-C1-C4-alkyl, -NHCO-phenyl, -NHSO2-C1-C4-alkyl, -NHSO2-phenyl, -SO2- C1-C4-alkyl and –SO2-phenyl, and R7 is hydrogen, C1-C6-alkyl, straight or branched, and which may be substituted with a phenyl ring which itself may also be substituted with one or two R10 radicals, and R8 is hydrogen, C1-C6-alkyl, straight or branched, which may be substituted with a phenyl ring which itself may also be substituted with one or two R10 radicals, and R9 is hydrogen, C1-C6-alkyl, straight or branched, which may also bear the substituent R16, or phenyl, pyridyl, pyrimidyl, pyridazyl, pyrazinyl, pyrazyl, naphthyl, quinolyl, imidazolyl, which may also bear the substituent R14 and R10 can be hydrogen, C1-C4-alkyl, branched or unbranched, -O-C1-C4-alkyl, OH, Cl, F, Br, J, SF3, NO2, NH2, CN, COOH, COO-C1-C4- alkyl, -NHCO-C1-C4-alkyl, -NHCO-phenyl, -NHSO2-C1-C4-alkyl, -NHSO2-phenyl, -SO2-C1-C4-alkyl and –SO2-phenyl, and R11 is hydrogen, C1-C6-alkyl, straight or branched, and which may be substituted with a phenyl ring which itself may also be substituted with one or two R10 radicals, and R 12 is hydrogen, C 1 -C 6 -alkyl, straight or branched, and [image] R13 is hydrogen, C1-C6-alkyl, branched or unbranched, which may also be substituted with a phenyl ring which may also carry the radical R10, and R14 is hydrogen, C1-C6-alkyl, branched or unbranched, -O-C1-C6-alkyl, branched or unbranched, OH, Cl, F, Br, J, SF3, NO2, NH2, CN, COOH, COO-C1 -C4-alkyl, or two radicals R14 can represent a bridge OC(R15)2O, i R15 is hydrogen, C1-C6-alkyl, branched or unbranched, and R16 can be a phenyl, pyridyl, pyrimidyl, pyridazyl, pyrazinyl, pyrazyl, pyrrolyl, naphthyl, quinolyl, imidazolyl ring, which can also carry one or two substituents R6, and A is –(CH2)m-, –(CH2)m-O-(CH2)o-, –(CH2)o-S-(CH2)m-, –(CH2)o-SO-(CH2)m-, –(CH2 )o-SO2–(CH2)m-, -CH=CH-, -C≡C-, -CO-CH=CH-, -(CH2)o-SO–(CH2)m-, –(CH2)m -NNSO–(CH2)o-, –(CH2)o-CONH–(CH2)o-, –(CH2)m-NHSO2–(CH2)o-, -NH-CO-CH=CH-, –(CH2 )m-SO2NN–(CH2)o-, -CH=CH-CONH- and [image] R1-A also represent together and V is phenyl, pyridines, pyrimidines, pyrazines, imidazoles and thiazoles, and x is 1, 2 or 3, and n is the number 0, 1 or 2, i m and o independently represent the number 0, 1, 2, 3 or 4. 2. Heterocyclic substituted amide of formula I according to claim 1, characterized in that V is pyridine or phenyl, and R 5 is hydrogen, and R9 is hydrogen, C1-C6-alkyl, branched or unbranched, which also carries the substituent R16, R16 is phenyl which may also carry one or two R14 substituents, i n is 0 and 1, i x is 1. 3. Heterocyclic substituted amide of formula I according to claim 1, characterized in that V is pyridine or phenyl, and R5 is CONR12R13, and R9 hydrogen, C1-C6-alkyl, branched or unbranched, which can also carry a substituent R16, R16 is phenyl which may also carry one or two R14 substituents, i n is 0 and 1, i x is 1. 4. Amide with heterocyclic substituents of formula I, according to claim 1, characterized in that V is pyridine or phenyl, and R2 is hydrogen, R 5 is hydrogen, and R9 hydrogen, C1-C6-alkyl, branched or unbranched, which can also carry the substituent R16, R16 phenyl which may also carry one or two R14 substituents, i n is 0 and 1, i x is 1. 5. Heterocyclic substituted amide of formula I according to claim 1, characterized in that V is pyridine or phenyl, and R2 is hydrogen, R5 is CONR12R13, and R9 is hydrogen, C1-C6-alkyl, branched or unbranched, can also carry the substituent R16, R16 is phenyl which may also bear one or two substituents R14, i n is 0 and 1, i x is 1. 6. Heterocyclic substituted amide of formula I according to claim 1, characterized in that A is –(CH2)m-, –(CH2)m-O-(CH2)o-, –(CH2)o-S-(CH2)m-, -CH=CH-, -C≡C-,–(CH2)m -NNSO–(CH2)o-, –(CH2)m-SO2NN–(CH2)o-, and B is pyridine or phenyl, and R 2 is hydrogen, and R 5 is hydrogen, and R9 is hydrogen, C1-C6-alkyl, branched or unbranched, which may also bear the substituent R16, and R 16 is phenyl, and m, n, o represent 0 and 1, i x is 1. 7. Heterocyclic substituted amide of formula I, according to the claim, characterized in that A is –(CH2)m-, –(CH2)m-O-(CH2)o-, –(CH2)o-S-(CH2)m-, -CH=CH-, -C≡C-,–(CH2)m -NNSO–(CH2)o-, –(CH2)m-SO2NN–(CH2)o-, and B is pyridine or phenyl, and R 2 is hydrogen, and R5 is CONR12R13, and R9 is hydrogen, C1-C6-alkyl, branched or unbranched, which may also bear the substituent R16, and R 16 is phenyl, and m, n, o represent 0 and 1, i x is 1. 8. Heterocyclic substituted amide of formula I according to claim 1, characterized in that V is pyridine or phenyl, and R1, R2 represent hydrogen, and R 5 is hydrogen, and R9 is hydrogen, C1-C6-alkyl, branched or unbranched, which may also bear the substituent R16, and R 16 is phenyl, and m, n, o represent 0, i x is 1. 9. Heterocyclic substituted amide of formula I according to claim 1, characterized in that V is pyridine or phenyl, and R1, R2 represent water, R5 is CONR12R13, and R9 is hydrogen, C1-C6-alkyl, branched or unbranched, which may also bear the substituent R16, and R 16 is phenyl, and m, n, o represent 0, i x is 1. 10. Use of amides of formula I according to claims 1-5, characterized in that they are used for the treatment of diseases. 11. Use of amides of formula I according to claims 1-5, characterized in that they are used as cysteine protease inhibitors. 12. Use according to claim 6, characterized in that they are used as inhibitors of cysteine proteases such as calpains and cathepsins, especially calpains I and II and cathepsins B and L. 13. Use of amides of formula I according to claims 1-5, characterized in that they are used for the production of drugs for the treatment of diseases in which increased calpain activity occurs. 14. Use of amides of formula I according to claims 1-5, characterized in that they are used for the production of drugs for the treatment of neurodegenerative diseases and neuronal injuries. 15. Use according to claim 9, characterized in that they are used for the treatment of those neurodegenerative diseases and neuronal injuries, which are caused by ischemia, trauma or extensive bleeding. 16. Use according to claim 10, characterized in that they are used for the treatment of stroke and craniocerebral trauma. 17. Use according to claim 10, characterized in that they are used for the treatment of Alzheimer's disease and Huntington's disease. 18. Use according to claim 10, characterized in that they are used for the treatment of epilepsy. 19. The use of compounds of formula I according to claims 1-5, characterized by the fact that they are used for the production of drugs and for the treatment of heart injuries after cardiac ischemia, kidney injury after renal ischemia, injury and reperfusion after vascular occlusion, skeletal and muscle injury, dystrophy muscles, injuries resulting from the proliferation of smooth muscle cells, coronary vasospasms, cerebral vasospasms, eye cataracts and restenosis of blood streams after angioplasty. 20. Use of amides of formula I according to claims 1-5, characterized in that they are used for the production of drugs for the treatment of tumors and their metastases. 21. Use of amides of formula I according to claims 1-5, characterized in that they are used for the production of drugs for the treatment of diseases in which elevated levels of interleukin 1 occur. 22. Use of amides of formula I according to claims 1-5, characterized in that they are used for the treatment of inflammation and rheumatic diseases. 23. Pharmaceutical preparation for oral, parenteral and intraperitoneal use, characterized in that in a single dose, apart from usual pharmaceutical excipients, it contains at least one amide I according to claims 1-5.