MXPA00009755A - New substituted amides, their production and their use - Google Patents

Abstract

The invention relates to amides of general formula (I) and their tautomeric and isomeric forms, their possible enantiomeric and diastereomeric forms and possible physiologically compatible salts, where the variables have the following meanings:R1 is C1-C6 alkyl, phenyl, naphthyl, quinolyl, pyridyl, pyrimidyl, pyridazyl, quinazolyl and quinoxalyl, whereby the rings can still be substituted with up to 2 R4 rests;R2 is -(CH2)m-R8, where R8 is phenyl, cyclohexyl or indolyl and m is 1 to 6;X is a bond, -CH2-, -CH2CH2-, -CH=CH-, -C=C-, -CONH-, -, -SO2NH-, and R1-X together can also be formula (a), R3 is hydrogen and CO-NR6R7;R4 is hydrogen, branched or unbranched C1-C4 alkyl and O-C1-C4-alkyl;R5 is hydrogen, branched or unbranched C1-C4 alkyl and O-C1-C4-alkyl;R6 is hydrogen, branched or unbranched C1-C6 alkyl;R7 is hydrogen, branched or unbranched C1-C6 alkyl;and n is 0, 1 or 2. The amides of formula (I) are inhibitors of enzymes, especially cysteine proteases such as calpains (calcium dependent cysteine proteases) and their isoenzymes and cathepsins, such as B and L.

Description

NOVEDOSAS AMIDAS SUBSTITUIDAS, ITS PREPARATION AND USE The present invention relates to novel amides, which are inhibitors of enzymes, in particular cysteine proteases, such as calpain (= calcium-dependent cysteine proteases) and their isoenzymes and cathepsins, for example B and L Calpains are intracellular proteolytic enzymes of the group so-called cysteine proteases and are found in many cells. The calpains are activated by an increased calcium concentration, a differentiation is made 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). Furthermore, calpain isoenzymes are currently postulated (K. Suzuki et al., Biol. Chem. Hoppe-Seyler, 1995, 376 (9), 523-9). It is suspected that calpains play an important part in various physiological processes. These include cleavage of regulatory proteins, such as protein kinase C, cytoskeletal proteins such as MAP 2 and spectrin, proteins, muscle, protein breakdown, in rheumatoid arthritis, proteins in platelet activation, neuropeptide metabolism, proteins in mitosis and others that are listed in M. J. Barrett et al., Life Sci. 1991, 48, 1659-69 and K.

K. ang et al., Trends in Pharmacol. Sci., 1994, 15, 412-9. Increased levels of calpain have been measured in various pathophysiological processes, for example in ischemia of the heart (for example, in cardiac infarction), of the kidney or central nervous system (for example an "attack"), inflammation, muscular dystrophy, cataracts of the eyes, injuries to the central nervous system (for example trauma), Alzheimer's disease, etc. (see K. K. Wang, earlier). A relationship of these diseases with increased and prolonged intracellular calcium levels is suspected. As a result, calcium-dependent processes become overactivated and are no longer subject to physiological regulation. Accordingly, the over activation of calpains can also initiate pathophysiological processes. Therefore, it was postulated that inhibitors of calpain enzymes may be useful for the treatment of these diseases. Various investigations confirm it. In this way, Seung-Chyul Hong et al., Stroke (Attack) 1994, 25 (3), 663-9 and R. T.

Bartus et al., Neurological Res. 1995, 17, 249-58, have shown a neuroprotective action of calpain inhibitors in acute neurodegenerative disorders or ischemia, such as after a stroke. Likewise, after experimental brain traumas, calpain inhibitors improve the recovery of deficits in memory power and neuromotor disorders that occur (K. E. Saatman et al. Proc. Nati.

Acad. Sci. USA, 1996, 93, 3428-3433). C. L. Edelstein et al., Proc. Na ti. Acad. Sci. USA, 1995, 92, 1662 -. 1662 - 6, found a protective action of t calpain inhibitors in kidneys damaged by hypoxia. Yoshida, Ken Ischi and collaborators, J'ap. Circ. J. 1995, 59 (1), 40-8, were able to show favorable effects of calpain inhibitors after cardiac damage that is caused by ischemia or reperfusion. Since calpain inhibitors inhibit the release of the β-AP4 protein, potential use as a therapeutic for Alzheimer's disease was proposed (J. Higaki et al., Neuron, 1995, 14, 651-59). The release of interleukin-1 is also inhibited by calpain inhibitors (N. Watanabe et al., Cytokine (Ci tocina) 1994, 6 (6), 597-601). In addition it was found that calpain inhibitors showed cytotoxic effects in tumor cells (E. Shiba et al., 20th Meeting Int. Ass. Breast Cancer Res., Sendai Jp, 1994, 25-28 Sept. , Int. J. Oncol. 5 (Suppl.), 1994, 381).

In addition, possible uses of calpain inhibitors are listed in K. K. Wang, Trends in Pha.rma.col. Sci. , 1994 , 412-8. Calpain inhibitors have already been described in the literature. These primordially however, are already irreversible or peptide inhibitors. As a rule, irreversible inhibitors are alkylating substances and have the disadvantage that they react non-selectively in the body or are unstable. In this way, these inhibitors often show undesirable side effects, such as toxicity, and accordingly they are restricted in their use or not usable. Irreversible inhibitors may include, for example, E-64 epoxides (E. B. McGowan et al., Biochem.

Biophys. Res. Commun. 1989, 158, 432-5), -haloketones (H.

Angliker et al., "Med. Chem. 1992, 35, 216-20) or disulfides (R. Matsueda et al., Chem. Let t. 1990, 191-194). Many known reversible inhibitors of cysteine proteases. such as calpain, are peptide aldehydes, in particular dipeptide and tripeptide aldehydes such as for example Z-Val-Phe-H (DL 28170) (S. Mehdi, Trends in Biol. Sci. 1991, 16, 150-3). Under physiological conditions, the peptide aldehydes have the disadvantage that they are often unstable considering the reactivity, they can be rapidly metabolized and are prone to non-specific reactions that can be the cause of toxic effects (J. A. Fehrentz and B. Castro, Synthesis (Synthesis) 1983, 676-78). In JP 08183771 (CA 1996, 605307) and in EP 520336, aldehydes which are derived from 4-piperidinoylamides and l-carbonylpiperidino-4-ylamides have been described as calpain inhibitors. However, the aldehydes claimed herein that are derived from heteroaromatically substituted amides of the general structure I have been previously described. Other aldehyde derivatives have been described by Chatterjee et al. Bioorgani c & Medicinal Chemistry Letters, 1997, 7, 287-290, Chatterjee et al. Bioorganic & Medicinal Chemistry Letters 1996, 6, 1619-1622, WO 97/10231 and WO 97/21690.

The peptide ketone derivatives are also inhibitors of cysteine proteases in particular calpain.

Thus, for example, in the case of cerin proteases, ketone derivatives are known as inhibitors, the keto group is activated by a group that removes electrons such as CF3 [sic]. In the case of cysteine proteases, derivatives with ketones activated by CF3 [sic] or similar groups are not very active or inactive (M. R.

Angelastro et al, J. "Med. Chem. 1990, 33, 11-13) Surprisingly, in the case of calpain to date only ketoha derivatives, where on the other hand, leaving groups in the alpha position cause an irreversible inhibition and on the other hand, a derivative of carboxylic acid activates the keto group, they were found as effective inhibitors (see MR Angelastro et al., see above; 92/11850, WO 92,12140, WO 94/00095 and WO 95/00535). However, of these ketoamides and keto esters, virtually only the peptide derivatives have been described as effective (Zhaozhao Li et al., J. Med. Chem. 1993, 36, 3472-80; SL Harbenson et al., J ". Med. Chem. 1994, 37, 2918-29 and, see above, MR Angelastro et al.). Only in Chatterjee et al. (See above) has a xanthene derivative of a ketobenzamide which has been described as calpain inhibitor. Ketobenzamides are already known in the literature. In this manner, the keto ester PhCO-Abu-COOCH2CH3 was described in WO 91/09801, WO 94/00095 and 92/11850. The analogous phenyl derivative Ph-CONH-CH (CH2Ph) -CO-COCOOCH3 was found by M. R. Angelastro et al., J. Med. Chem. 1990, 33, 11-13 however, it is only a weak calpain inhibitor. This derivative is also described by J. P. Burkhardt, Tetrahedron Lett. , 1988, 3433-36. The significance of these substituted benzamides, however, is never investigated so far. In a number of therapies, such as attack, the active compounds are administered intravenously as an infusion solution. 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. Many of the described calpain inhibitors however have the disadvantage that they only show a small solubility or are not soluble in water and thus are not suitable for intravenous administration. Active compounds of this type can only be administered using auxiliaries that are intended to impart water solubility (see R.T. Bartus et al., J. Cereb. Blood Flow Metab. 1994, 14, 537-544). These auxiliaries, for example polyethylene glycol, frequently nevertheless have side effects or are even intolerable. A non-peptide calpain inhibitor which according to this is water soluble without auxiliaries therefore can probably be administered with better acceptability, thus has a great advantage. Highly effective non-peptide calpain inhibitors that have sufficient solubility in water previously have not been described and therefore would be novel.

In the present invention, non-peptide aldehydes, keto carboxylic acid esters and keto amide derivatives are described. These compounds are novel and surprisingly show the possibility of obtaining powerful non-peptide inhibitors of cysteine proteases such as for example calpain, by incorporating rigid structural fragments. In addition, in the case of the present compounds of the general formula I, which all carry at least one aliphatic amine radical, salt bonds with acids are possible. This leads to an improved solubility in water and therefore the compounds show the desired profile for intravenous administration, as is necessary for example in attack therapy. The present invention relates to substituted amides of the general formula I and its tautomeric and isomeric forms, possible enantiomeric and diastereomeric forms, as well as possible physiologically tolerable salts, wherein the variables have the following meanings: Rl [sic] can be alkyl with 1 to 6 carbon atoms, phenyl, naphthyl, quinolyl, pyridyl, pyrimidyl, pyridazyl, quinazolyl and quinoxalyl, wherein the rings may additionally be substituted by up to two radicals R4 [sic], and R2 [sic] is - (CH2) m-R8 [sic], wherein R8 [sic] may be phenyl, cyclohexyl- [sic] or indolyl and m = 1 to 6, and X is a bond -CH2-, -CH2CH2-, -CH = CH-, -C = C-, -CONH-, - S02NH- [sic], and [sic] Rl-X [sic] together are also R3 [sic] is hydrogen and CO-NR6R7 [sic], R4 [sic] is hydrogen, alkyl having 1 to 4 carbon atoms [sic], which is unbranched or branched and -O-alkyl having 1 to 4 carbon atoms carbon [sic]; R5 [sic] is hydrogen, alkyl having 1 to 4 carbon atoms [sic], which is unbranched or branched and -O-alkyl having 1 to 4 carbon atoms [sic]; R6 is hydrogen, alkyl having 1 to 6 carbon atoms, which is unbranched or branched and R7 [sic] is hydrogen, alkyl having 1 to 6 carbon atoms, which is unbranched or branched and n is [sic] a number 0, 1 or 2. The compounds of the formula I can be used as racemates, as enantiomerically pure compounds or as diastereomers. If enantiomerically pure compounds are desired, they can be obtained, for example, by carrying out a classical racemate resolution with the compounds of the formula I or their intermediates using a convenient optically active acid or base. On the other hand, enantiomeric compounds can also be prepared by using commercially available compounds, for example optically active amino acids such as phenylalanine, tryptophan and tyrosine. The present invention also relates to compounds which are mesomeric or tautomeric with the compounds of the formula I, for example those in which the aldehyde or keto group of the formula I is present as an enol tautomer. The present invention also relates to the physiologically tolerable salts of the compounds I, which can be obtained by reacting the compounds I with a suitable base or acid. Acids and convenient bases are listed, for example, in Fortschritte der Arzneimittelforschung [Advances in Drug Research], 1996, Birkhuser Verlag, Vol. , pgs. 224-285. 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. The amides I according to the invention which carry an aldehyde group can be prepared in various forms which have been established in synthesis scheme 1. > Synthesis scheme 1 R2 R2 IV 1. - Hydrolysis 2.- Oxidation R2 R2 1. - NH (CH3) OH 2.- Deprotection 1.- Hydrolysis 2.- Reduction R2 Reduction Vil VI R2 The carboxylic acids II are bonded with suitable amino alcohols III to give the corresponding amides IV. This is done using methods of Peptide coupling, which are mentioned either in C. R. Larock, Comprehensive Organic Transformations, VCH Publisher, 1989, p. 972f. or in Houben-Weyl, Methoden der organischen Chemie [Methods of Organic Chemistry], 4a. Edition, E5, Chapter V. The preferred reaction is carried out using "activated" acid derivatives of II, the acidic COOH group is converted to a COL group. L is a leaving group such as for example Cl, imidazole or N-hydroxybenzotriazole. This activated acid is then converted to the amides IV using amines. The reaction is carried out in anhydrous inert solvents such as methylene chloride, tetrahydrofuran and dimethylformamide at temperatures of -20 to + 250C [sic]. These carboxylic acid esters IV (R '= O-alkyl) are converted to the acids IV (R '= H) using acids such as trifluoroacetic acid [sic] or hydrochloric acid or bases such as lithium hydroxide, sodium hydroxide or potassium hydroxide, in aqueous medium or in mixtures of water and organic solvents such as alcohols and tetrahydrofuran at room temperature or elevated temperatures, such as 25-100 ° C. These IV derivatives (R '= H) that are obtained can be oxidized in aldehyde derivatives I according to the invention. It is possible to use different reactions of oxidation, for this (see CR Larock, Comprehensive Organic Transformations, VCH Publisher, 1989, page 604f.) Such as, for example, similar oxidations Swern and Swern (TT Tidwell, Synthesis (Synthesis) 1990, 857 -70), sodium hypochloride [sic] / TEMPO (SL Harbenson et al., See above) or Dess -Martin (J. Org. Chem. 1983, 48, 4155). Preferably, the reaction here is carried out in inert aprotic solvents such as dimethylformamide, tetrahydrofuran or methylene chlorine using oxidants such as DMSO / p and x S03 or DMSO / oxalyl chloride at temperatures from -50 to + 250C [sic], depending on the method (see previous references). Alternatively, the carboxylic acid II can be reacted with amino aminohydroxamic acid derivatives VI to give benzamides VII. In this case, the same reaction procedure is used as in the IV preparation. The hydroxamic derivatives [lacuna] VI are obtained from the protected amino acids V by reaction with a hydroxylamine. In this process, an amide preparation process that has already been described is also employed. The removal of the protective group X, for example Boc, is carried out in a conventional manner, for example using trifluoroacetic acid. The amidohydroxamic acids VII thus obtained can be converted to the acids I in accordance with the invention by reduction. In this process, for example, lithium aluminum hydride is used as a reductant at temperatures of -60 to 00 C [sic] in inert solvents such as tetrahydrofuran or ether. Analogous to the latter process, carboxylic acids or acid derivatives such as esters IX (Y = COOR ', COSR') can also be prepared which can also be converted into the aldehydes I according to the invention by reduction. These processes are listed in R. C. Larock, Comprehensive Organic Transor- mations, (Complete Organic Transformations) VCH Publisher, 1989, p. 619-26. The preparation of the substituted amides I according to the invention, [lacuna] transport a ketoamide group or ketoester, can be carried out in various forms that have been established in synthesis scheme II.

Synthesis scheme 2 The carboxylic acids II are reacted with amino hydroxy carboxylic acid derivatives X (for the preparation of XI see S. L. Harbenson et al., J.

Med. Chem. 1994, 37, 2918-29 or J. P. Burkhardt et al. Tetrahedron Lett. 1988, 29, 3433-3436) under usual peptide coupling methods (see above, Houben-Weyl), the amides XIII are obtained. These carboxylic acid esters XIII (R '= O-alkyl) are converted to the acids XIII (R' = H) using acids such as trifluoroacetic acid or hydrochloric acid or bases such as lithium hydroxide, sodium hydroxide or potassium hydroxide in aqueous medium or in mixtures of water and organic solvents such as alcohols or tetrahydrofuran at room temperature or elevated temperatures, such as 25-100 ° C. The derivatives XIII obtained can be oxidized to the keto carboxylic acid derivatives I 'according to the invention. Various usual oxidation reactions can be used for this (see CR Larock, Comprehensive Organic Transformations, VCH Publisher, 1989, page 604 f.) Such as, for example, Swern and Swern analogue oxidations, preferably dimethyl complex. sulfoxide / pyridine-sulfur trioxide, in solvents such as methylene chloride or tetrahydrofuran, if appropriate with addition of dimethyl sulfoxide at room temperature or temperatures of -50 to 25 ° C (TT Tidwell, Synthesis (Synthesis) 1990, 857-70) or sodium hypochlorite [sic] / TEMPO (S. L. Harbenson et al., See above). If XI are a-hydroxy esters (X = O-alkyl), and these can be hydrolysed in carboxylic acids XII, the reaction is carried out analogously to the above methods, but preferably using lithium hydroxide in water / mixtures. tetrahydrofuran at room temperature, it being necessary to take into consideration however that in this case for the protective group R ', a The radical is chosen such as, for example, tert-butyl O, which allows the selective cleavage of one of the two ester groups. The preparation of other amides XIII is carried out by reaction with amines under coupling conditions that have already been described. The alcohol derivative XIII can be oxidized again to give keto carboxylic acid derivatives I 'according to the invention. The preparation of the carboxylic acid esters II has already been described in some cases or is carried out according to usual chemical methods. Compounds wherein X is a bond, are prepared by usual aromatic coupling, for example Suzuki coupling, with boric acid derivatives and halides under palladium catalysis or copper catalyzed coupling of aromatic halides. Bridged alkyl radicals (X = - (CH 2) m-) can be prepared by reduction of the analogous ketones or by alkylation of the lithium organ, for example ortho-phenyloxazolidines, or other organometallic compounds (cf. Dordor et al., J. Chem Soc. Perkin Trans. I, 1984, 1247-52). The bridged alkene and alkyl compounds are prepared, for example, by the Heck reaction from suitable aromatic halides and alkenes and alkyls (see I. Sakamoto et al., Chem. Pharm. Bull., 1986, 34, 2754-59).

Amides and sulfonamides are prepared from the amines and acid derivatives, analogously to the methods described above. Alternatively, compounds of the general formula I can also be synthesized by modifying or exchanging the reaction sequences listed in schemes 1 and 2. In this way, for example a sulfonamide I (RlX [sic] = RS02NH) can be prepared from a derivative IV ^ RlX [sic] = N02) by reducing the nitro group to the amine catalytically in a usual manner using hydrogen in a catalyst, such as palladium / carbon, and then reacting the resulting amine with a sulfonyl chloride to give an IV derivative (RlX [sic] = RS02NH). Additional reaction to give I is carried out as illustrated in the scheme, by ester hydrolysis and oxidation. Analogously, intermediaries IV and XI (RlX [sic] (chemical groups such as nitro, amino, halogen etc.), can be converted into derivatives where RlX [sic] correspond to additional radicals mentioned in the general claim. The reactions are carried out here analogously to the processes described above or analogously to general or usual methods. The heterocyclic substituted amides I contained in the present invention are inhibitors of cysteine proteases, in particular cysteine proteases such as calpains I and II and cathepsins B and L. The inhibitory activity of the heterocyclicly substituted amides I, is determined using enzyme tests usual in the literature, a concentration of the inhibitor at which 50% of the enzyme activity is inhibited ( = IC50 [sic]), is determined as an action scale. The amides I were measured in this manner for inhibitory activity of calpain I, calpain II and cathepsin B. Cathepsin B test The inhibition of cathepsin B is determined analogously to a method by S. Hasnain et al., J. Biol. Chem. 1993, 268, 235-40. 2 μL of an inhibitor solution prepared from the inhibitor and DMSO (final concentrations: 100 μM to 0.01 μM) are added to 88 μL of cathepsin B (cathepsin) B of human liver (Calbiochem), diluted to 5 units in 500 μM buffer). This mixture is preincubated at room temperature (25 ° C) for 60 minutes and the reaction is then initiated by the addition of 10 μL of 10 mM Z-Arg-Arg-pNA (in buffer 10% DMSO). The reaction is checked at 405 nM in a microtiter plate reader for 30 minutes. IC50s [sic] is then determined from the maximum gradients. Calpain test I and II The test for inhibitory properties of calpain inhibitors is carried out in buffer using 50 mM tris HCl, pH 7.5; 0.1 M NaCl; 1 mM dithiothreitol [sic]; 0.11 mM CaCl2, the fluorogenic calpain substrate Suc-Leu-Tyr-AMC (25 mM dissolved in DMSO, Bachem / Switzerland), is used. Human μ-calpain is isolated from erythrocytes and after several chromatographic steps (DEAE-Sepharose, phenyl-Sepharose, Superdex 200 and Blue Sepharose), the enzyme has a purity of > 95%, estimated according to SDS-PAGE, Western drying and N-terminal sequencing is obtained. The fluorescence of the cleavage product 7-amino-4-methylcoumarin (AMC) is verified in a Spex-Fluorolog fluorometer at? Ex = 380 nm and? Em = 460 nm. In a measurement range of 60 min, the substrate cleavage is linear and the autocatalytic activity of calpain is low if experiments are carried out at temperatures of 12 ° C. The inhibitors and calpain substrate are added to the experimental batch as DMSO solutions, where DMSO should not exceed 2% in the final concentration. In an experimental batch, 10 μl of the substrate (250 μM final) and then 10 μl of μ-calpain (2 μg / ml final, i.e. 18 nM) are added to a 1 ml cuve containing buffer. The; Calpain-mediated cleavage of the substrate is measured for 15-20 min. 10 μl of inhibitor (50-100 μM solution in DMSO) is then added and the inhibition of cleavage is measured for an additional 40 minutes. The K values are determined according to the classical equation for reversible inhibition: Ki [sic] = 1 / (VQ / V - 1, where I = inhibitory concentration, v0 = initial velocity before addition of the inhibitor, vx = velocity of equilibrium reaction Velocity is calculated from v = AMC release / time, ie height / time i Calpain is an intracellular cysteine protease Calpain inhibitors must pass through the cell membrane in order to avoid the breakdown of intracellular proteins by calpain Some known calpain inhibitors, such as E 64 and leupeptin, only cross the cell membranes with difficulty and accordingly show, although they are good calpain inhibitors, only a poor action The goal is to find compounds that have better access to the membrane.As a demonstration of the membrane access of calpain inhibitors, human platelets are used s Decomposition mediated by tyrosine kinase calpain pp60src [sic] in platelets. After activation of platelets, tyrosine kinase pp60s c is cleaved by calpain. This was investigated in detail by Oda et al., in J. Biol.

Chem. , 1993, 268, 12603-12608. It was shown in this context that the cleavage of pp60src [sic] can be avoided by calpeptin, a calpain inhibitor. The cellular effectiveness of our substances was tested following this publication. Fresh human blood treated with citrate, centrifuged at 200 g for 15 minutes. Platelet-rich plasma was harvested and diluted 1: 1 with platelet buffer (platelet buffer: 68 mM NaCl, 2.7 mM KCl, 0.5 mM MgCl2 x 6 H20, 0.24 mM NaH2P04 x H20, 12 mM NaHCO3, 5.6 mM glucose, 1 mM EDTA, pH 7.4). After a centrifugation and washing step with platelet buffer, the platelets were adjusted to 107 [sic] cells / ml. The isolation of human platelets was carried out at RT. In the test batch, isolated platelets (2 x 106 [sic]) were pre-incubated at 37 ° C with different concentrations of inhibitors (dissolved in DMSO) for 5 minutes. The platelets were then activated with ionophore A23187 1 μM and 5 mM CaCl2. After incubation for 5 minutes, the platelets were centrifuged briefly at 13000 and the nodule was collected in SDS i sample buffer (SDS sample buffer: tris 20 mM HCl, 5 mM EDTA, 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 in a 12% concentration gel and pp60src [sic] and their cleavage products of 52 kDa and 47 kDa were identified by Western drying. The anti-Cis-src polyclonal rabbit antibody (pp60src [sic]) used was purchased from the company Biomol Feinchemikalien (Hamburg). This primary antibody is detected using a second antibody coupled with goat HRP (Boehringer Mannheim, FRG). Western drying was carried out according to known methods. The quantification of cleavage of pp60src [sic] is carried out by densitometry, the controls used are platelets not activated (control 1: without excision) and platelets treated with ionophore and calcium (control 2: corresponds to 100% excision). The ED50 value corresponds to the concentration of the inhibitor at which the intensity of the color reaction is reduced by 50%. Cell death induced by glutamate in cortical neurons The test is carried out as Choi D. W., Maulucci-Gedde M.A. and Kriegstein A.R., "Glutamate neurotoxicity in cortical cell culture" (Neurotoxicity of glutamate in cortical cell culture). J ". Neurosci 1989, 7, 357-368. The cortex moieties of mouse embryos at 15 days of age were dissected and the individual cells were obtained enzymatically (trypsin). These cells (neurons, glia and cortical) are inoculated in 24-well plates. After three days (plates coated with laminin) or 7 days (plates coated with ornithine), the mitosis treatment is carried out using FDU (5-fluoro-2-deoxyuridine). 15 days after cell preparation, cell death is induced by addition of glutamate (15 minutes). After removing the glutamate, the calpain inhibitors are added. 24 hours later, cell damage is determined by determination of hydrogenase lactate (LDH) in the cell culture supernatant. It is postulated that calpain also plays a part 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 an additional model, cell death is induced with calcium in the presence of calcium ionophore in a human cell line. Calpain inhibitors must pass into the cell and inhibit calpain in order to prevent induced cell death. Calcium-mediated cell death in cells NT2. Cell death can be induced in a human NT2 cell line by calcium in the presence of ionophore A 23187. 105 [sic] cells / wells were coated on microtiter plates 20 hours before the experiment.

After this period, the cells were incubated with various concentrations of inhibitors in the presence of 2.5 μM ionophore and 5 mM calcium. 0.05 ml of XTT (cell proliferation kit II, Boehringer Mannheim) are added to the reaction batch after 5 hours. The optical density is determined approximately 17 hours later, according to the manufacturer's instructions, on the Easy Reader EAR 400 from the company SLT. The optical density at which half of the cells died is calculated from the two controls with cells without inhibitors, which were incubated in the absence and presence of ionophore. In a number of neurological diseases or psychological disorders, increased glutamate activity occurs, leading to states of over stimulation or toxic effects in the central nervous system (CNS = Central Nervous System). Glutamate mediates its effects by various receptors. Two of these receptors are classified by the specific agonists NMDA receptor and AMPA receptor. Antagonists against these glutamate-mediated effects can thus be used for the treatment of these diseases, in particular for therapeutic administration against neurodegenerative diseases such as Huntington's disease and Parkinson's disease, neurotoxic disorders after hypoxia, anoxia, ischemia and subsequent injuries, such as they occur after Attack and trauma, or alternatively as antiepileptics (cf. Arzneim, Forschung 1990, 40, 511-514, TIPS, 1990, 11, 334-338, Drugs of the Future (Drugs of the future) 1989, 14, 1059-1071). The protection against over-stimulation by excitatory amino acids (NMDA antagonism or AMPA in mice) As a result of the intracerebral administration of excitatory amino acids (EAA), this is induced on a massive stimulus, which in a short time leads to spasms and death of animals (mice). These symptoms can be inhibited by systemic administration, for example intraperitoneal of centrally active compounds (EAA antagonists). Since the excessive activation of EAA receptors of the central nervous system plays an important part in the pathogenesis of various neurological disorders, the conclusion can be drawn from the EAA antagonism demonstrated in vivo with respect to a possible therapeutic utility of the substances against CNS disorders of this type. As a measure of the efficacy of the substances, an ED50 value is determined in which 50% of the animals are free of symptoms as a result of a fixed dose of either NMDA or AMPA as a result of the i.p. previous of the standard substance ..

The heterocyclic substituted amides I are inhibitors of cysteine derivatives such as calpain I or II and cathepsin B or L and thus can be used for the control of diseases that are associated with increased enzymatic activities of the enzymes calpain or cathepsin enzymes. The present amides I according to this can be used for the treatment of neurodegenerative diseases that occur after ischemia, reperfusion damage after vascular occlusion trauma, subarachnoid hemorrhages and attacks, and neurodegenerative diseases, such as dementia by multiple infarction, Alzheimer's disease, Huntington's disease and epilepsies and also for the treatment of heart damage after cardiac ischemia, damage to the kidneys after renal ischemia, skeletal muscle damage, muscular dystrophies, damage that occurs due to cell proliferation of smooth muscle, coronary vasospasm, cerebral vasospasm, cataracts in the eyes, restenosis of bloodstream after angioplasty. Still further, the amides I may be useful in chemotherapy of tumors and metastases thereof and for the treatment of diseases where an increased level of interleukin-1 occurs, such as in disorders of inflammation and rheumatism. In addition to the usual pharmaceutical auxiliaries, The pharmaceutical preparations according to the invention contain a therapeutically effective amount of the compounds I. For local external application, for example in powders, ointments or sprays, the active compounds can be contained in the usual concentrations. As a rule, the active compounds are contained in an amount of 0.001 to 1% by weight, preferably 0.001 to 0.1% by weight. In the case of internal administration, the preparations are administered in individual doses. 0.1 a 100 mg are provided in a single dose per kg of body weight. The preparation can be administered daily in one or more doses depending on the nature and severity of the disorders. According to the desired type of administration, the pharmaceutical preparations according to the invention contain the usual excipients and diluent in addition to the active compound. For local external application, pharmaceutical auxiliaries such as ethanol, isopropanol, ethoxylated castor oil, hydrogenated ethoxylated castor oil, polyacrylic acid, polyethylene glycol, polyethylene glycol stearate, ethoxylated fatty alcohols, paraffin oil, petrolatum, and lanolin. For internal administration, for example, they are suitable lactose, propylene glycol, ethanol, starch, talc and polyvinylpyrrolidone. . Antioxidants such as tocopherol and butylated hydroxyanisole as well as butylated hydroxytoluene, flavor improving additives, stabilizers, emulsifiers and lubricants may additionally be contained. The substances contained in the preparation in addition to the active compound and the substances used in the production of the pharmaceutical preparations are toxicologically acceptable and compatible with the respective active compound. The pharmaceutical preparations are prepared in the usual manner, for example by mixing the active compound with other customary excipients and diluents. The pharmaceutical preparations can be administered in various administration methods, eg, orally, parenterally such as intravenously by subcutaneous intraperitoneal infusion and topically. In this way, preparation forms such as tablets, emulsions, infusion and injection solutions, pastes, ointments, gels, creams, lotions, powders and sprays are possible. Examples Example 1 (S) -3-Carboxy-5- (2-naphthylsulfonamido) -N- (3-phenylpropan-1-al-2-yl) benzamide a) (S) -3- (Ethoxycarbonyl-5-nitro-N- ( 3-phenylpropan-l-ol-2-yl) -benzamide 10 g (41.8 mmoles) of monoethyl 5-nitroisophthalate, 6.3 g (41.8 mmoles) of (S) -phenylalaninol and 10.6 g (104.5 mmoles) of triethylamine were dissolved in 200 ml of methylene chloride at room temperature and stirred for 1.9 minutes 1.9 g (13.9 mmoles) of 1-hydroxybenzotriazole and subsequently in portions, 8 g (41.6 mmoles) of (N-dimethylaminopropyl) -N-ethylcarbodiimide The whole mixture is stirred at room temperature for 16 'hours.The reaction batch is diluted with methylene chloride to twice the volume, using methylene chloride and washed successively with 2M hydrochloric acid, water , 2M sodium hydroxide solution and water again The organic phase was separated, dried and concentrated in vacuo. 9.1 g of the intermediary are obtained. b) (S) -5-amino-3-ethoxycarbonyl-N- (3-phenylpropan-l-ol-2-yl) -benzamide 9 g (24.3 mmoles) of the intermediate are dissolved in 300 ml of ethanol and hydrogenated after of addition of 1 g of palladium / carbon (10% concentration). In reaction batches, it was then filtered and the filtrate was concentrated in vacuo. 8.1 g of the intermediate was obtained. c) (S) -3-Ethoxycarbonyl-5- (2-naphthylsul-fonamido) -N- (3-phenyl-propan-1-ol-2-yl) -benzamide 2 g (5.84 mmol) of intermediate Ib and 2.4 ml (17.4 mmoles) of triethylamine were dissolved in 50 ml of tetrahydrofuran. A solution of 1.32 g (5.82 mmoles) of 2-naphthalenesulfonyl chloride in 30 ml of tetrahydrofuran is then added dropwise at 0 ° C and the reaction batch is then stirred at 40 ° C for 8 hours. The reaction batch is then concentrated in vacuo and the residue partitioned between water and ethyl acetate. The ethyl acetate phase was further washed with 2M hydrochloric acid and water and then dried and concentrated in vacuo. The residue thus obtained which is purified by chromatography on silica gel (eluent: methylene chloride / ethanol = 20/1), 0.65 g i of the intermediate, is obtained, d) (S) - 3 -Carboxi -5- (2-naphthi sulphonamido) -N- (3-phenylpropan-l-ol-2-yl) benzamide 0.65 g (1.2 mmoles) of the intermediate is dissolved in 30 ml of tetrahydrofuran and treated with 0.15 g (6.3 mmoles) of lithium hydroxide, dissolved in 15 ml of water. The whole mixture is stirred at room temperature for 26 hours. The organic solvent is then removed under vacuum and the aqueous residue is acidified using 2M hydrochloric acid. The precipitate obtained was filtered off with suction and dried. 0.46 g of the intermediate is obtained, d) (S) - 3 - Carboxy - 5 - (2-naphthi sulphonamido) -N- (3-phenylpropan-1-al-2-yl) benzamide 0.46 g (0.91 g) mmoles) of the intermediate compound Id and 0.37 g (3.65 mmol) of triethylamine are dissolved in 10 ml of dry dimethyl sulfoxide and treated with 0.44 g (2.76 mmoles) of pyridine-sulfur trioxide complex. The whole mixture is stirred at room temperature for 16 hours. The reaction mixture is then added to ice-water, acidified with 1M hydrochloric acid and the precipitate is filtered off with suction. 0.36 g of the product is obtained. NMR- ^ H (D6-DMSO): d = 2.9 (1H), 3.3 (1H), 4.5 (1H), 7-2 (5H), 7.6-7.9 (5H), 8.0-8.2 (4H), 8.5 (2H), 9.1 (1H), 9.6 (1H) and 10.9 (1H) ppm. Example 2 N- (l-Carbamoyl-2-oxo-4-phenylpropan-2-yl) -3-carboxy-5- (2-naphthylsulfonamido) benzamide N- (1-Carbamoyl-2-hydroxy-4-phenylpropan-2-yl) -3-ethoxycarbonyl-5-nitrobenzamide 5.2 g (21.7 mmoles) of monoethyl-5-nitroisophthalate, 5 g (21.7 mmoles) of 3- amino-2-hydroxy-3-phenylbutyramide and 11.2 g (110.5 mmoles) of triethylamine are dissolved in 200 ml of methylene chloride at room temperature and stirred for 30 minutes. 2.9 g (21.6 mmoles) of 1-hydroxybenzotriazole and subsequently in portions 4.6 g (22.8 mmoles) of (N-dimethyl-aminopropyl) -N-ethylcarbodiimide is then added with cooling with ice. The whole mixture is stirred at room temperature for 16 hours. The reaction batch is diluted to twice the volume using Methylene chloride and washed successively with 2M hydrochloric acid, water, 2M sodium hydroxide solution and water again. The organic phase was separated, dried and concentrated in vacuo. 2.6 g of the intermediate are obtained. b) 5-Amino-N- (l-carbamoyl-2-hydroxy-4-phenylpropan-2-yl) -3-ethoxycarbonylbenzamide 2.6 g (6.25 mmoles) of intermediate 2a are dissolved in 50 ml of dimethyl formamide, diluted with 200 ml of ethanol and hydrogenated after the addition of 1 g of palladium / carbon (10% in concentration). The reaction batch is then filtered and the filtrate concentrated in vacuo. 1.8 g of the intermediate is obtained. c) N- (1-Carbamoyl-2-hydroxy-4-phenylpropan-2-yl) -3-ethoxycarbonyl-5- (2-naphthylsulfonamido) benzamide 1.8 g (4.7 mmoles) of intermediate 2b and a tip filled with 4-dimethylaminopyridine spatula is dissolved in 30 ml of pyridine. 1.2 g (5.1 mmol) of naphthalenesulfonyl chloride are added dropwise at room temperature and the whole mixture is further stirred for 16 hours. The reaction batch is then emptied on ice-water and acidified with 2M hydrochloric acid. This aqueous phase is extracted several times with ethyl acetate. The combined organic phases were dried and concentrated in vacuo. He The residue thus obtained was further treated successively with ether and a little ethyl acetate, 1.3 g of the intermediate is obtained, d) N- (l-Carbamoyl-2-hydroxy-4-phenylpropan-2-yl) -3-carboxy -5- (2-naphthylsulfonamido) benzamide 1.25 g (2.2 mmoles) of intermediate 3c are dissolved in 10 ml of tetrahydrofuran and treated with 0.21 g (8.7 mmoles) of lithium hydroxide, dissolved in 50 ml of water. The whole mixture is stirred at room temperature for one hour. The organic solvent is then removed under vacuum and the aqueous residue is acidified with 2M hydrochloric acid. The formed precipitate is filtered off with suction and dried. 1.0 g of the intermediary is obtained. e) N- (1-Carbamoyl-2-oxo-4-phenylpropan-2-yl) -3-carboxy-5- (2-naphthylsulfonamido) benzamide 0.9 g (1.6 mmoles) of intermediate 2d and 1.4 ml (9.9 mmoles) ) of triethylamine are dissolved in 25 ml of dry dimethyl sulfoxide and treated at room temperature with 0.78 g (4.9 mmoles) of pyridine sulfur trioxide complex dissolved in 13 ml of dimethyl sulfoxide. The whole mixture is stirred at room temperature for one hour. The reaction mixture is then emptied onto ice-water and acidified with 1M hydrochloric acid and the precipitate is filtered off with suction, 0.59 g of the product is obtained. NMR-H1 (CF3, COOD): d = 2.9 (1H), 3.1 (1H), 5.3 (1H), 7-0-8-3 (17H), 8.4 (1H) and 9.1 (1H) ppm. The following examples can be prepared analogously to the above procedures; R2 Example R 'Rz R3 RXX 2 3 -COOH (CH2) 3CH3 CONH2 5-Naphth-2-yl-S02NH 3 3 -COOH, CH2Ph H 5-Phenyl-S02NH 4 3 -COOH CH2Ph CONH2 5-Phenyl-S02NH 3 -COOH CHa h H 5-n-Bu il-S02NH 6 3 -COOH CH2Ph CONH2 5-n-Butyl-S02NH 7 4 -COOH CH2Ph H 3-Phenyl-S02NH 8 4 -COOH CH2Ph H 5 -Naft-2-yl-S02NH 9 4 -COOH CH2Ph CONH2 5 -Naft-2-yl -S02NH 4 -COOH CH2Ph H 3 -CH = CH-Phenyl 11 4 -COOH CH2Ph H 3- (Pyrid-2-yl) 12 3 -COOH, CH2Ph H 4 -CH = CH-Phenyl If R3 = H, then the configuration of the C2 atom is (S); in the case of R3 = CONH2, this is (R, S) (Threw CM LD THE

Claims (2)

1. - An amide of the general formula I and its tautomeric and isomeric forms, possible enantiomeric and diastereomeric forms, as well as possible physiologically tolerable salts, wherein the variables have the following meanings: Rl [sic] can be alkyl having 1 to 6 carbon atoms, phenyl, naphthyl, quinolyl, pyridyl, pyrimidyl, pyridazil, quinazolyl and quinoxalyl, wherein the rings may additionally be substituted by up to two radicals R4 [sic], and R2 [sic] is - (CH2) m-R8 [sic], wherein R8 [sic] may be phenyl, cyclohexyl- [sic] or indolyl and m = la 6, and X is a bond -CH2-, -CH2CH2-, -CH = CH-, -C = C-, -CONH-, -S02NH- [sic], and [sic] Rl-X [sic] together are also and R3 [sic] is hydrogen and CO-NR6R7 [sic], R4 [sic] is hydrogen, alkyl with 1 to 4 carbon atoms [sic], which is unbranched or branched and -O-alkyl with 1 to 4 atoms carbon [sic]; R5 [sic] is hydrogen, alkyl having 1 to 4 carbon atoms [sic], which is unbranched or branched and -O-alkyloxy with 1 to 4 carbon atoms [sic]; R6 [sic] is hydrogen, alkyl having 1 to 6 carbon atoms, which is unbranched or branched and R7 [sic] is hydrogen, alkyl having 1 to 6 carbon atoms, which is unbranched or branched and n is a number 0 , 1 or 2. 2. An amide of the formula I according to claim 1, characterized in that R1 [sic] is phenyl, naphthyl, butyl and quinolyl, R2 [sic] is benzyl and R3 [sic] is hydrogen and X is S02NH and R4 [sic] is hydrogen. 3. An amide of the formula I according to claim 1, characterized in that R1 [sic] is phenyl, naphthyl, butyl and quinolyl, R2 [sic] is benzyl and R3 [sic] is C0NH2 and X is S02NH and R4 [sic] sic] is hydrogen. 4. The use of amides of the formula I according to claims 1 to 3, for the treatment of diseases. 5. The use of amides of the formula I according to claims 1 to 3, as inhibitors of cysteine proteases. 6. - Use in accordance with the claim 5, as inhibitors of cysteine proteases such as calpains and cathepsins, in particular calpains I and II and cathepsins B and L. 7. The use of amides of the formula I according to claims 1 to 3, for the production of pharmaceuticals for the treatment of diseases where increased calpain activity occurs. 8. The use of amides of the formula I according to claims 1 to 3, for the production of pharmaceuticals for the treatment of neurodegenerative diseases and neuronal damage. 9. - Use in accordance with the claim 8, characterized for the treatment of those neurodegenerative diseases and neuronal damage that are caused by ischemia, trauma or nosebleeds. 10. - Use in accordance with the claim 9, for the treatment of stroke and craniocerebral trauma. 11.- The use in accordance with the claim 9, for the treatment of Alzheimer's disease and Huntington's disease. 12. The use according to claim 9, characterized for the treatment of epilepsy. 13. The use of compounds of the formula I of according to claims 1 to 3, for the production of pharmaceuticals and treatments for damage to the heart after cardiac ischemia, damage to the kidneys after renal ischemia, reperfusion injury after vascular occlusion, skeletal muscle damage, muscular dystrophies, damage which results due to the proliferation of smooth muscle cells, coronary vasospasm, cerebral vasospasm, cataracts of the eyes and restenosis of bloodstreams after angioplasty. 14. The use of the amides of the formula I according to claims 1 to 3, for the production of pharmaceuticals for the treatment of tumors and their metastases. 15. The use of the amides of the formula I according to claims 1 to 3, for the production of pharmaceuticals for the treatment of diseases where increased levels of interleukin-1 occur. 16. The use of the amides according to claims 1 to 3, for the treatment of immunological diseases such as inflammations and rheumatic disorders. 17. A pharmaceutical preparation for oral parenteral and intraperitoneal use, comprising by dose individual, in addition to the usual pharmaceutical auxiliaries, at least one amide I [sic] according to claims 1 to 3. SUMMARY OF THE INVENTION Amides of the. general formula I and its tautomeric and isomeric forms, possible enantiomeric and diastereomeric forms, as well as possible physiologically tolerable salts, wherein the variables have the following meanings: Rl [sic] can be alkyl having 1 to 6 carbon atoms, phenyl, naphthyl, quinolyl, pyridyl, pyrimidyl, pyridazil, quinazolyl and quinoxalyl, wherein the rings can additionally be substituted by up to two radicals R4 [sic], and R2 [sic] is - (CH2) "V.-R8 [sic], wherein R8 [ sic] can be phenyl, cyclohexyl- [sic] or indolyl and m = 1 to 6, and X is a bond -CH2-, -CH2CH2-, -CH = CH-, -C = C-, -CONH-, -S02NH- [sic], and [sic] Rl-X [sic] j untos are also and R3 [sic] is hydrogen and C0-NR6R7 [sic], R4 [sic] is hydrogen, alkyl with 1 to 4 carbon atoms [sic], which is unbranched or branched and -O-alkyl with 1 to 4 atoms carbon [sic]; R5 [sic] is hydrogen, alkyl having 1 to 4 carbon atoms [sic], which is unbranched or branched and -O-alkyl having 1 to 4 carbon atoms [sic]; R6 is hydrogen, alkyl having 1 to 6 carbon atoms, which is unbranched or branched and R7 [sic] is hydrogen, alkyl having 1 to 6 carbon atoms, which is unbranched or branched and n is a number 0, 1 or

2. 18772