DE10114762A1 - Treatment of lung diseases, especially adult respiratory distress syndrome, by administration of cysteine protease inhibitor, especially calpain inhibitor - Google Patents

Abstract

The use of cysteine protease inhibitor(s) (I) in the production of pharmaceutical compositions for treating lung diseases is new.

Description

The present invention relates to the use of cysteine pro tease inhibitors, especially calpain inhibitors for the manufacture treatment of a pharmaceutical agent for the treatment of disease lung, especially Adult Respiratory Distress Syndrome (ARDS).

Calpaine are a family of calcium-activated, cytosolic Cy stone proteases. The classic Calpaine consist of an iso shape-specific, catalytic and mostly calcium-binding large subunit (80 kDa), Capn1-Capn3 and Capn5-Capn12, and an immutable small subunit (30 kDa) that Capn4 is called (N. Dear et al., Genomics 1997, 45: 175-184). A summary of the currently known members of the The mammalian calpaine gene family is available on the Internet (http: // Ag.Arizona.Edu/calpains).

Calpaine cleave numerous substrates, such as protein Kinase A and C, cytoskeletal proteins such as MAP 2 and spectrin, Membrane proteins, such as EGF-R, transcription factors, G proteins, Muscle proteins, proteins in rheumatoid arthritis, in AK activation of platelets and in neuropeptide metabolism and Proteins in mitosis. So the Calpainen play a role in egg attributed to a variety of physiological processes, for example wise in apoptosis (K. K. Wang, Trends Neurosci. 2000, 23: 20-26), cell division and growth regulation (R.L. Mellgren, Bio chem. Biophys. Res. Commun. 1997, 236: 555-558), modulation of the Interactions of the integrin cytoskeleton (S. M. Schoenwaelder et al., J. Biol. Chem. 1997, 272: 1694-1702) and synaptic pla sticity (S.L. Chan and M.P. Mattson, J. Neurosci. Res. 1999, 58: 167-190).

They were also treated with various pathophysiological pro associated with increased calcium levels gel and a possibly resulting increased calpain Activity and / or elevated calpain levels are characterized are. Therefore, it has been postulated that inhibitors of calpain enzymes can be useful for the treatment of diseases. Ver different possible areas of application of calpain inhibitors such as for example ischemia of the heart (e.g. heart attack), the Kidney or central nervous system (e.g. stroke), inflammation exercises, muscular dystrophies, cataracts of the eyes, injuries to the  Central nervous system, Alzheimer's disease, Huntington's disease, Demyelination, epilepsy, etc. are described in K.K. Wang et al., Trends in Pharmacol. Sci. 1994, 15: 412-419; E. Carafoli and M. Molinari, Biochem. Biophys. Res. Commun. 1998, 247: 193-203; H. Sorimachi et al., Biochem. J. 1997, 328: 721-732; and K.K. Wang and P. W. Yuen, Adv. Pharmacol. 1997, 37: 117-152. The utility of such inhibitors for the treatment of crane kung lung is not mentioned.

In the lungs, Capn1 (µ-Calpain) and Capn2 (m-Calpain) are ex primed. Capn1 or µ-calpain is determined by µ-molar concentrations activated by calcium ions while Capn2 or m-calpain, the most abundant calpain isoenzyme in the lungs of the mouse, by m molar concentrations of calcium ions is activated (P. John son, int. J. Biochem. 1990, 22 (8): 811-22; A. Miller et al., To xicology 1995, vol. 97, 141-149). Calpaine are in the lungs u. a. involved in surfactant secretion from alveolar type II cells (U. J. Zimmerman et al., Cell Calcium 1995, 18 (1): 1-8).

E. J. Blumenthal and A. M. Malkinson, Arch. Biochem. Biophys. 1987, Vol. 256, 1: 19-28, describe a decrease in the cytosoli calpain activity in the repair phase after a complete butylated hydroxytoluene (BHT) induced, reversible lungs damage. Although the inhibibility of a membrane-associated Pro tease by the calpain inhibitors calpastatin, leupeptin and E-64 was observed, the authors obviously do not see any Occasion, the use of calpain inhibitors for treatment propose. The authors believe that there are more Studies required to understand the physiological role of Calpain to determine and a relationship between change gene calpain activity and cellular regulatory mechanism men to manufacture.

A.C. K. Miller et al., Toxicology 1995, vol. 97, 141-149 below seek the role of Capn2 in BHT-induced lung damage in the mouse. It increases in the first 40 minutes after BHT administration Capn2 level on, but drops shortly afterwards below the starting level veau from. The reduced Capn2 concentration is linear the extent of the damage. The level of pulmonary protein Ki nose C-α (PKC-α) is believed to be influenced by Capn2 in decreased the first 45 minutes after BHT administration. For this reason PKC-α and Calpain levels have been suggested as early Biomar ker for the extent of alveolar damage. The car Although they suspect that the observed changes play a role could play in triggering the alveolar damage that exact meaning of the transient rise in the Calpain level and the calpain  However, concentration remains unclear. Although calpain inhibitors were already known at the time, their application for the treatment of the injuries obviously not in Er weighed.

Polytrauma, sepsis, intoxication and burns often cause complications, which in the worst case can lead to death from acute lung failure (ARDS, adult re spiratory distress syndrome) or multiple organ failure via an event cascade. ARDS results in respiratory insufficiency and the resulting hypoxemia and insufficient oxygen supply to the organism. The lungs are mostly affected first and especially after sepsis, trauma and burns. For example, polymorphonucleolar granulocytes (PMN) are activated and attached to the capillary endothelial wall. The activated PMN then secrete mediators, proteases and O ₂ radicals and migrate into the interstitium and the alveolar space. This is followed by hemodynamic changes, increased permeability and inflammatory processes, and finally to interstitial and alveolar edema and the development of ARDS.

By application of activated PMN can also in the iso perfused lungs trigger edema. It continues Neuhof and Fritz, Prog. Clin. Biol. Res. 1989, 308: 305-314, the Elastase secreted by PMN plays an essential role because by the elastase inhibitor Eglin c in the experiment a PMN-indu can prevent decorated pulmonary edema. However, none of this can be done Indication of a possible role of calpaines in pathogenesis of a pulmonary edema.

The object of the present invention was to develop new ways of treating lung (preventive and / or curative) diseases of the lungs, especially ARDS.

Surprisingly, the above task could be done by the user development of cysteine protease inhibitors, in particular calpain-inhi bittern, for the production of a pharmaceutical agent for loading treatment of diseases of the lungs. With invent Diseases according to the invention are in particular a crane lung problems caused by sepsis, skin burns, Trauma and / or intoxication are triggered, such as in particular ARDS.

The term "treatment" as used herein includes the Thera Pie of an existing disease with the invention pharmaceutical agents, as well as the prophylaxis of such a disease,  d. H. preventive administration according to the invention pharmaceutical agent.

The term "diseases of the lungs" as used here summarizes all diseases and conditions of the lungs, d. H. all acutely triggered, acquired, chronic and genetically related functional disorders of the lungs, their origin and / or course a calpain, especially Capn1 and Capn2 involved is.

All known cysteine are for use in accordance with the invention Suitable tease inhibitors, the cysteine proteases such as wise Cathepsin B, L, S, K, F and V and especially Calpaine inhibit. Calpain inhibitors are preferred, particularly preferred are inhibitors of such calpains, which are expressed in the lungs such as Capn1 and Capn2 in particular. Particularly preferred one uses calpain inhibitors that are found in suitable medicines have the preparations administered orally or parenterally, and especially those that are in a pharmaceutically acceptable Solvents are soluble.

Numerous cysteine protease inhibitors have been described in the literature. A general distinction is made between reversible and irreversible inhibitors. The irreversible inhibitors include, for example, the epoxides E-64 (EB McGowan et al., Biochem. Biophys. Res. Commun. 1989, 158: 432-5), α-halogenetones, diazoketones (H. Angliker et al. , J. Med. Chem. 1992, 35: 216-20) or disulfides (R. Matsueda et al., Chem. Lett. 1990, 191-194). Irreversible inhibitors are usually alky lating substances such as Z-Leu-Val-Gly-CHN ₂ , Z-Gly-Leu-PhCH ₂ Cl ₂ and have the disadvantage that they are unstable, react non-selectively in the organism and therefore undesirable Show side effects and the effect, as mentioned above, is difficult to control.

The irreversible cysteine protease inhibitors also include the peptidic ketones which carry a leaving group in the α-position, such as, for example, peptidic ketoamides or keto esters, e.g. B. the ketoester PhCO-Abu-COOCH ₂ CH ₃ (see MR Angelastro et al., See above; WO-A-92/11850; WO-A-92,12140; WO-A-94/00095; WO-A -95/00535; Zhaozhao Li et al., J. Med. Chem. 1993, 36: 3472-80; SL Harbenson et al., J. Med. Chem. 1994, 37: 2918-29; JP Burkhardt, Tetrahedron Lett ., 1988; 3433-36). The leaving group in the α-position can be a hetero group, e.g. B. an amino or amido group (DL Flynn et al. J. Am. Chem. Soc. 1997, 119: 4874-4881; S. Natarajan et al., J. Enzym. Inhib. 1988, 2: 91-97; JD Godfrey et al., J. Org. Chem. 1986, 51: 3073-3075; GB-A-2170200; EP-A-159156; EP-A-132304; US-A-4470973 and JP-A-59033260) , Examples of sulfur and oxygen derivatives are described in EP-A-603873, WO-A-95/15749 and RE Dolle et al., J. Med. Chem. 1995, 38: 220-222. In Chatterjee et al. (See above) a xanthine derivative of a ketobenzamide is also described as a cysteine protease inhibitor.

Reversible cysteine protease inhibitors include peptides cal aldehydes, especially dipeptic and tripepidic Aldehydes such as Z-Val-Phe-H (MDL 28170) (S. Mehdi, Trends in Biol. Sci. 1991, 16: 150-3), AK 275 and AK 295, as well N-acetyl-leucyl-leucyl-norleucinal or N-acetyl-leucyl-leucyl methional, which inhibit selective Capn1 and Capn2. However peptidic aldehydes, as well as the peptidic ketone derivatives, unstable due to their reactivity (J.A. Fehrentz and B. Cas tro, Synthesis, 1983, 676-678). There are therefore only a few peptides aldehydes and ketones used as active ingredients in the treatment of Diseases used are known. To the peptidic Cal pain inhibitors continue to count calpeptin, calpain inhibitor I. and II and the endogenous calpain inhibitors leupeptin and calpa statin, with only calpastatin selectively inhibiting Capn1 and Capn2, while the other peptide inhibitors are comparatively un are selective and therefore only limited to therapeutic Suitable use (K.K. Wang et al., Trends in Pharmacol. Sci. 1994, 15: 412-419).

Become reversible non-peptide cysteine protease inhibitors in JP-A-8183759, JP-A-8183769, JP-A-8183771 and EP-A-520336 wrote. These are derived from dipeptides Aldehydes, with saturated carbocyclic rings, e.g. B. Cyclohe xane, or saturated heterocyclic rings, e.g. B. piperidines put an amino acid in peptide cysteine protease inhibito be installed. JP-A-8183771 (CA 1996, 605307) and the EP-A-520336, for example, describe aldehydes which differ from Piperidin-4-ylcarboxamides and 1-carbonylpiperidin-4-ylcar derive bonsäureamiden as cysteine protease inhibitors. Other Aldehyde derivatives are described in Chatterjee et al., Bioorganic & Medi cinal Chemistry Letters, 1997, 7: 287-290, Bioorganic & Medicinal Chemistry Letters 1996, 6: 1619-1622, WO-A-97/10231 and WO-A-97/21690.

Particularly preferred cysteine protease inhibitors for the indication according to the invention are heterocyclic substituted amides of the general formula I, described in WO 99/61423, WO 99/54304, WO 99/54305, WO 99/54294, WO 99/54320, WO 99/54293 , DE-A-196 48 793, DE-A-196 42 591 and WO 99/54310, to which express reference is made here,

and their tautomeric and isomeric forms, enantiomers and diastereomeric forms, as well as their physiologically compatible Salts, in which the variables are those in the following connecting group have defined meanings.

Group I according to WO 99/61423

Compounds of formula I, wherein
R ¹ for

stands in what
B represents phenyl, naphthyl, pyridyl, pyrimidyl, pyrazyl, pyridazyl, quinolyl, quinazyl, quinoxalyl, thienyl, benzothienyl, benzofuranyl, furanyl or indolyl,
A for a condensed cycle selected from

stands in what
n represents 0, 1 or 2, and
R ⁴ for hydrogen, (CH ₂ ) _m NR ⁸ R ⁹ , O (CH ₂ ) _m NR ⁸ R ⁹

stands,
wherein
R ^{8 stands} for C ₁ -C ₆ alkyl, which can be substituted with a phenyl ring, where the phenyl ring itself can be substituted with one or two radicals R ¹⁰ ,
in which
R ¹⁰ for C ₁ -C ₄ alkyl, -OC ₁ -C ₄ alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, CONH ₂ , COOH, COO-C ₁ -C ₄ alkyl, -NHCO-C ₁ -C ₄ alkyl, -NHCO-phenyl, -NHSO ₂ -C ₁ -C ₄ alkyl, -NHSO ₂ -phenyl, -SO ₂ -C ₁ -C ₄ - Is alkyl or -SO ₂ phenyl;
R ^{9 represents} C ₁ -C ₆ alkyl which may be substituted with a phenyl ring, where the phenyl ring itself may also be substituted by one or two R ¹⁰ radicals;
R ^{15 represents} hydrogen or R ⁸ as defined above; and
m represents 1, 2, 3, 4, 5 or 6;
Y is hydrogen, C ₁ -C ₆ alkyl, OC ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkylphenyl, C ₂ -C _6- alkenyl-phenyl, C ₂ -C ₆ -alkynyl-phenyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ -C ₄ -alkyl, NHCO -C ₁ -C ₄ alkyl, NHCO-phenyl, NHSO ₂ -C ₁ -C ₄ alkyl, NHSO ₂ phenyl, SO ₂ -C ₁ -C ₄ alkyl, SO ₂ phenyl or CONHR ¹¹ ,
wherein
R ^{11 represents} hydrogen or C ₁ -C ₆ alkyl;
and
p represents 0, 1 or 2;
R ^{2 represents} C ₁ -C ₆ alkyl, which can also carry a phenyl, cyclohexyl, pyridyl, thienyl, indolyl or naphthyl ring, the ring itself being substituted with up to two radicals Y as defined above can be and
R ^{3 represents} hydrogen, COOR ⁵ or CO-Z, wherein
R ^{5 represents} C ₁ -C ₆ alkyl which can be substituted with a phenyl ring, where the ring itself can also be substituted with one or two radicals R ¹⁰ ,
Z for NR ⁶ R ⁷

stands in what
R ^{6 represents} hydrogen or C ₁ -C ₆ alkyl,
R ^{7 represents} hydrogen or C ₁ -C ₆ alkyl, which can also be substituted by a phenyl or pyridine ring, which can optionally carry a radical R ¹⁰ , or by

can be substituted
o represents 0, 1, 2, 3 or 4 and R ⁸ , R ⁹ and R ^{15 have} the meaning given above.

Group II according to WO 99/54304

Compounds of formula I, wherein
R ¹ for

stands in what
p represents 0, 1 or 2,
B represents an unsaturated heterocyclic ring selected from pyridine, pyrimidine, pyrazine, imidazole and thiazole;
A for chlorine, bromine, fluorine, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkenyl, C ₁ -C ₆ alkynyl, C ₁ -C ₆ alkylphenyl, C ₁ -C ₆ alkenyl- Phenyl, C ₁ -C ₆ alkynylphenyl, phenyl, NHCO-C ₁ -C ₄ alkyl, NHSO ₂ -C ₁ -C ₄ alkyl, -NHCOPhenyl, -NHCO-naphthyl, NO ₂ , -OC ₁ - C ₄ alkyl or NH ₂ , where the aromatic rings can also carry one or two radicals R ⁵ , and two radicals A together can also represent a chain -CH = CH-CH = CH- and thus form a fused benzo ring , which in turn can be substituted with an R ⁵ ,
in which
R ⁵ for C ₁ -C ₄ alkyl, -OC ₁ -C ₄ alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ -C ₄ -Alkyl, -NHCO-C ₁ -C ₄ -alkyl, -NHCO-phenyl, -NHSO ₂ -C ₁ -C ₄ -alkyl, -NHSO ₂ -phenyl, -SO ₂ -C ₁ -C ₄ -alkyl, - (CH ₂ ) _n -NR ¹² R ¹³ or -SO ₂ -phenyl,
Y represents phenyl, naphthyl, quinolyl, pyridyl, pyrimidyl, pyrazyl, pyridazyl, imidazolyl, thiazole, quinazyl, iso quinolyl, quinazyl, quinoxalyl, thienyl, benzothienyl, benzofuranyl, furanyl or indolyl, the rings still having up to 3 radicals R. ⁵ may be substituted according to the above definition,
x for a bond, - (CH ₂ ) _m -, - (CH ₂ ) _m -O- (CH ₂ ) _o -, - (CH ₂ ) _o -S- (CH ₂ ) _m , - (CH ₂ ) _o -SO- (CH ₂ ) _m -, - (CH ₂ ) _o -SO ₂ - (CH ₂ ) _m -, -CH = CH-, -CbC-, -CO-CH = CH-, (CH ₂ ) _o -CO- (CH ₂ ) _m -, - (CH ₂ ) _m -NHCO (CH ₂ ) _o -, - (CH ₂ ) _m -CONH- (CH ₂ ) _o -, - (CH ₂ ) _m -NHSO ₂ - (CH ₂ ) _o -, -NHCO-CH = CH-, - (CH ₂ ) _m -SO ₂ NH- (CH ₂ ) _o -, -CH = CH-CONH- or

stands,
in the case of CH = CH double bonds both the E and the Z form can be present; and
m and o are independently 0, 1, 2, 3 or 4;
or
X and Y together, if appropriate, also for

wherein
R ^{5 has} the meaning given above and
n represents 0, 1 or 2;
R ^{2 represents} -C ₁ -C ₆ alkyl and optionally a SCH ₃ -, phenyl, cyclohexyl, cycloheptyl, cyclopentyl, indolyl, pyridyl or naphthyl radical, which in turn can carry up to two radicals R ⁵ can be substituted as defined above,
R ^{3 represents} hydrogen, COOR ⁶ or CO-Z, wherein
R ^{6 represents} hydrogen or C ₁ -C ₆ alkyl, which can be substituted by a phenyl ring, which itself can also be substituted by one or two R ⁹ radicals, where
R ⁹ for hydrogen, C ₁ -C ₄ alkyl, -OC ₁ -C ₄ alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ - C ₄ alkyl, -NHCO-C ₁ -C ₄ alkyl, -NHCO-phenyl, -NHSO ₂ -C ₁ -C ₄ alkyl, -NHSO ₂ -phenyl, -SO ₂ -C ₁ -C ₄ alkyl or -SO ₂ phenyl; and
Z for NR ⁷ R ⁸ ,

stands,
wherein
R ^{10 represents} hydrogen or C ₁ -C ₆ alkyl which can be substituted by a phenyl ring, where the phenyl ring itself can also be substituted by up to two radicals R ⁹ as defined above;
R ^{7 represents} hydrogen or C ₁ -C ₆ alkyl; and
R ^{8 stands} for hydrogen or C ₁ -C ₆ alkyl, which can also be substituted with a phenyl ring, where the phenyl ring can also carry a radical R ⁹ as defined above, and with

can be substituted, wherein
R ^{10 has} the meaning given above;
R ^{11 represents} hydrogen or C ₁ -C ₆ alkyl which can be substituted by a phenyl ring, where the phenyl ring itself can also be substituted by up to two radicals R ⁹ as defined above, and
n stands for 0, 1, 2, 3 or 4.

Group III according to WO 99/54305

Compounds of formula I, wherein
R ¹ for

stands in what
x represents 0, 1 or 2,
B represents a phenyl, pyridine, pyrimidine, pyrazine or pyridazine ring;
A represents piperazine, homopiperazine, hexahydroazepine, piperidine or pyrrolidine and can also carry a radical R ⁵ ,
in which
R ^{5 stands} for C ₁ -C ₆ alkyl, which may also have a substituent R ¹¹ , or for a phenyl, pyridyl, pyrimidyl, pyridazyl, pyrazinyl, pyrazyl, naphthyl, thienyl, piperidinyl, pyrrolidinyl or Imida zyl ring, which ring can also carry one or two substituents R ⁶ , wherein
R ⁶ for C ₁ -C ₆ alkyl, OC ₁ -C ₆ alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ -C ₄ - Alkyl or C ₁ -C ₄ alkyl-NR ⁹ R ¹³ or two radicals R ^{6 can form} a bridge OC (R ⁹ ) ₂ O.
in which
R ^{9 represents} hydrogen or C ₁ -C ₆ alkyl; and
R ^{13 represents} hydrogen, a C ₁ -C ₄ -alkyl chain or C ₀ -C ₄ -alkylphenyl-, where the phenyl ring can also carry one or two radicals R ¹² , where
R ¹² for C ₁ -C ₆ alkyl, OC ₁ -C ₆ alkyl, ver branched or unbranched, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH or COO- C ₁ -C ₄ alkyl; and
R ^{11 represents} a phenyl, pyridyl, pyrimidyl, naphthyl, thienyl, furyl, pyridazyl, pyrazinyl, pyrazyl, pyrrolyl or imidazyl ring, this ring also having one or two substituents R ⁶ according to the above Can carry definition;
and
Y and Z independently of one another are hydrogen, C ₁ -C ₆ -alkyl, OC ₁ -C ₆ -alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ -C ₄ alkyl, NHCO-C ₁ -C ₄ alkyl, NHCO-phenyl, CONHR ⁹ , NHSO ₂ -C ₁ -C ₄ alkyl, NHSO ₂ phenyl, SO ₂ -C ₁ -C ₄ alkyl or SO ₂ phenyl; or Y and Z together can form a chain -CH = CH-CH = CH- which can also carry one or two substituents R ⁶ as defined above;
R ^{2 represents} C ₁ -C ₆ alkyl, which may be a SCH ₃ radical or a cyclohexyl, cyclopentyl, cycloheptyl, phenyl, pyridyl, pyrimidyl, pyridazyl, pyrazyl, indolyl, thienyl - Or wear naphthyl ring, where the rings can be substituted with up to two radicals R ⁷ , wherein
R ⁷ for C ₁ -C ₄ alkyl, OC ₁ -C ₄ alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ -C ₄ - Alkyl, CONHR ⁹ , NHCO-C ₁ -C ₄ alkyl, NHCO-phenyl, NHSO ₂ -C ₁ -C ₄ alkyl, NHSO ₂ phenyl, SO ₂ -C ₁ -C ₄ alkyl or SO ₂ phenyl stands,
in which
R ^{9 has} the meaning given above and
R ^{3 represents} hydrogen or -COR ⁸ , where R ^{8 represents} -OR ⁹ or -NR ⁹ R ¹⁰ , wherein
R ^{9 has} the meaning given above and
R ^{10 represents} hydrogen or C ₁ -C ₆ alkyl, which may also be substituted with a phenyl ring, the phenyl ring may also carry a radical R ¹² as defined above, and with

may be substituted, wherein R ^{13 has} the meaning indicated Be.

Group IV according to WO 99/54294

Compounds of formula I, wherein
R ¹ for

stands in what
A represents C ₁ -C ₆ alkyl, phenyl, naphthyl, quinolyl, pyridyl, pyrimidyl, pyridazyl, quinazolyl or quinoxalyl, it being possible for the rings to be substituted with up to two radicals R ⁴ , where
R ^{4 represents} hydrogen, C ₁ -C ₄ alkyl or -OC ₁ -C ₄ alkyl;
X represents a bond, -CH ₂ -, -CH ₂ CH ₂ -, -CH = CH-, -C∼C-, -CONH- or SO ₂ NH-, or
AX together for

stand in what
R ^{5 represents} hydrogen, C ₁ -C ₄ alkyl or -OC ₁ -C ₄ alkyl; and
n represents 0, 1 or 2;
R ^{2 represents} - (CH ₂ ) _m -R ⁸ , wherein
R ^{8 represents} phenyl, cyclohexyl or indolyl, and
m represents a number from 1 to 6, and
R ^{3 represents} hydrogen or CO-NR ⁶ R ⁷ , wherein
R ^{6 represents} hydrogen or C ₁ -C ₆ alkyl, and
R ^{7 represents} hydrogen or C ₁ -C ₆ alkyl.

Group V, according to WO 99/54320

Compounds of formula I, wherein
R ¹ for

stands,
x represents 1, 2 or 3,
B represents phenyl, pyridine, pyrimidine, pyrazine, imidazole or thiazole and
A for - (CH ₂ ) _m -, - (CH ₂ ) _m -O- (CH ₂ ) _o -, - (CH ₂ ) _o -S- (CH ₂ ) _m -, - (CH ₂ ) _o -SO - (CH ₂ ) _m , - (CH ₂ ) _o -SO ₂ - (CH ₂ ) _m , -CH = CH-, -CbC-, -CO-CH = CH-, (OH ₂ ) _o -CO- ( CH ₂ ) _m -, - (CH ₂ ) _m NHCO- (CH ₂ ) _o -, - (CH ₂ ) _m -CONH- (CH ₂ ) _o -, - (CH ₂ ) _m -NHSO ₂ - (CH ₂ ) _o -, -NH-CO-CH = CH-, - (CH ₂ ) _m -SO ₂ NH- (CH ₂ ) _o - or -CH = CH-CONH-,
where m and o are independently 0, 1, 2, 3 or 4;
Y represents hydrogen, C ₁ -C ₆ -alkyl, phenyl, naphthyl, quinolinyl, pyridyl, pyrimidyl, pyrazyl, pyridazyl, chinazolyl, quinoxalyl, thienyl, benzothienyl, benzofuranyl, furanyl or indolyl, the rings still containing up to 3 R ⁶ radicals can be substituted;
or
YA together also for

stand in what
R ⁶ for hydrogen, C ₁ -C ₄ alkyl, -OC ₁ -C ₄ alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ - C ₄ alkyl, -NHCO-C ₁ -C ₄ alkyl, -NHCO-phenyl, -NHSO ₂ -C ₁ -C ₄ alkyl, -NHSO ₂ -phenyl, -SO ₂ -C ₁ -C ₄ alkyl or -SO ₂ phenyl; and
n represents 0, 1 or 2; and
R ⁴ for hydrogen, C ₁ -C ₆ alkyl, OC ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkyl phenyl, C ₂ - C ₆ -alkenyl-phenyl, C ₂ -C ₆ -alkynyl-phenyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ -C ₄ -alkyl, NHCO-C ₁ -C ₄ alkyl, NHCO-phenyl, CONHR ⁹ , NHSO ₂ -C ₁ -C ₄ alkyl, NHSO ₂ phenyl, SO ₂ -C ₁ -C ₄ alkyl or SO ₂ phenyl, in which
R ^{9 represents} hydrogen, C ₁ -C ₆ alkyl, which can also carry a substituent R ¹⁶ , phenyl, pyridyl, pyrimidyl, pyridazyl, pyrazinyl, pyrazyl, naphthyl, quinolinyl or imidazolyl, each of these rings having one or two substituents R ^{14 can} wear, being
R ^{16 represents} a phenyl, pyridyl, pyrimidyl, pyridyl, pyrazinyl, pyrazyl, pyrrolyl, naphthyl, quinolinyl or imidazolyl ring which can also carry one or two substituents R ⁶ ; and
R ¹⁴ for C ₁ -C ₆ alkyl, OC ₁ -C ₆ alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH or COO-C ₁ -C ₄ - Alkyl is or two radicals R ^{14 can form} a bridge OC (R ¹⁵ ) ₂ O, wherein
R ^{15 represents} hydrogen or C ₁ -C ₆ alkyl, and
R ^{5 represents} NR ⁷ R ⁸ or a ring, selected from

in which
R ⁶ and n have the meanings given above,
R ^{7 represents} hydrogen or C ₁ -C ₆ alkyl, which may be substituted by a phenyl ring, which phenyl ring itself may also be substituted by one or two radicals R ¹⁰ , where
R ¹⁰ for C ₁ -C ₄ alkyl, -OC ₁ -C ₄ alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ -C ₄ -Alkyl, -NHCO-C ₁ -C ₄ -alkyl, -NHCO-phenyl, -NHSO ₂ -C ₁ -C ₄ -alkyl, -NHSO ₂ -phenyl, -SO ₂ -C ₁ -C ₄ -alkyl or - SO ₂ phenyl;
R ^{8 represents} hydrogen or C ₁ -C ₆ alkyl, which can be substituted by a phenyl ring, this phenyl ring itself can also be substituted by one or two radicals R ¹⁰ as defined above,
R ^{2 represents} -C ₁ -C ₆ alkyl, which can also carry a phenyl, pyridyl, thienyl, cyclohexyl, indolyl or naphthyl ring, this ring having up to two radicals R ^{6 in} accordance with the above Definition can be substituted, and
R ^{3 is} hydrogen, COOR ¹¹ or CO-Z, where Z is NR ¹² R ¹³ , wherein
R ^{11 represents} hydrogen or C ₁ -C ₆ alkyl, which may be substituted by a phenyl ring, this phenyl ring itself may also be substituted by one or two radicals R ¹⁰ as defined above, and
R ^{12 is} hydrogen, C ₁ -C ₆ alkyl,

stands, wherein R ⁷ and R ^{8 have} the meaning given above be; and
R ^{13 represents} hydrogen or C ₁ -C ₆ -alkyl, which can also be substituted by a phenyl ring, this phenyl ring can also carry a radical R ¹⁰ as defined above, and by

can be substituted.

Group VI according to WO 99/54293

Compounds of formula I, wherein
R ¹ for

stands,
wherein
Y for chlorine, bromine, fluorine, C ₁ -C ₆ alkyl, NHCO-C ₁ -C ₄ alkyl, NHSO ₂ -C ₁ -C ₄ alkyl, NO ₂ , -OC ₁ -C ₄ alkyl, CN , COOH, CONH ₂ , COO-C ₁ -C ₄ -alkyl, SO ₂ -C ₁ -C ₄ -alkyl, -SO ₂ Ph, SO ₂ NH-C ₁ -C ₄ -alkyl, iodine, SO ₂ NH ₂ or NH ₂ ;
A represents an aromatic or heteroaromatic ring such as naphthyl, quinolyl, quinoxyl, benzimidazolyl, benzothienyl, quinazolyl, phenyl, thienyl, imidazolyl, pyridyl, pyrimidyl or pyridazyl, the ring also having at least one radical R ⁹ and / or with up to two radicals R ⁸ can be substituted, where
R ^{8 is} C ₁ -C ₄ alkyl, -OC ₁ -C ₄ alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COOC ₁ -C ₄ alkyl , -NHCO-C ₁ -C ₄ alkyl, phenyl, NHCO-phenyl, -NHSO ₂ -C ₁ -C ₄ alkyl, -NHSO ₂ -phenyl, -SO ₂ -C ₁ -C ₄ alkyl, pyridine or SO ₂ phenyl, and
R ^{9 represents} (CH ₂ ) _p -R ¹² ,
in which
R ¹² for pyrrolidine, morpholine, piperidine, hexahy droazepine, homopiperazine

is in which R ^{10 is} -C ₁ -C ₆ alkyl, branched or unbranched, and can also carry a phenyl ring which in turn is optionally substituted with up to two radicals R ¹¹ , where R ^{11 is} hydrogen, C ₁ -C ₄ -Alkyl, branched or unbranched, -OC ₁ -C ₄ -alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ -C ₄ -alkyl, NHCO- C ₁ -C ₄ alkyl, -NHSO ₂ -C ₁ -C ₄ alkyl or -SO ₂ -C ₁ -C ₄ alkyl;
R ^{13 represents} hydrogen or C ₁ -C ₆ alkyl, branched or unbranched, and
p represents 0, 1 or 2; and
B for a bond, - (CH ₂ ) _m -, - (CH ₂ ) _m -O- (CH ₂ ) _o -, - (CH ₂ ) _o -S- (CH ₂ ) _m -, - (CH ₂ ) _o -SO- (CH ₂ ) _m -, - (CH ₂ ) _o -SO ₂ - (CH ₂ ) _m , -CH = CH-, -CbC-, -CO-CH = CH, - (CH ₂ ) _o -CO- (CH ₂ ) _m , - (CH ₂ ) _m -NHCO- (CH ₂ ) _o -, - (CH ₂ ) _m -CONH- (CH ₂ ) _o -, - (CH ₂ ) _m -NHSO ₂ - (CH ₂ ) _o -, -NH-CO-CH = CH- or - (CH ₂ ) _m -SO ₂ NH- (CH ₂ ) _o -, where
m and o are independently 0, 1, 2, 3 or 4;
or
AB together also for

are in which R ⁸ and R ^{9 have} the meaning given above and z is 0, 1 or 2;
R ^{2 represents} -C ₁ -C ₆ alkyl, where one of the C atoms in this chain can be substituted with a phenyl ring, cyclohexyl ring, indolyl ring and / or a SCH ₃ group and the phenyl ring in turn may optionally be substituted with up to two radicals R ⁴ may be substituted, where R ^{4 is} hydrogen, C ₁ -C ₄ -alkyl, -OC ₁ -C ₄ -alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ -C ₄ alkyl or NHCO-C ₁ -C ₄ alkyl; and
R ^{3 represents} CH ₂ -NR ⁵ CO-R ⁶ or CH ₂ -NHR ⁵ SO ₂ -R ⁶ ,
wherein
R ^{5 represents} hydrogen or C ₁ -C ₄ alkyl; and
R ^{6 represents} hydrogen, phenyl, naphthyl or C ₁ -C ₆ alkyl, it being possible for a C atom in the chain to be substituted with a phenyl ring, this phenyl ring itself also being substituted with one or two radicals R ⁴ as defined above can be.

Group VII according to DE-196 48 793

Compounds of formula I, wherein
R ¹ for

stands,
wherein
n represents 1 or 2,
A for hydrogen, chlorine, bromine, fluorine, phenyl, a phenyl optionally substituted by a C ₁ -C ₄ hydrocarbyl radical, -NHCO-C ₁ -C ₄ alkyl, -NHCOPh, -NHCO-naphthyl, -NHSO ₂ - C _1-4 alkyl, -COO-C _1-4 alkyl, -CONH ₂ , COOH, -OC _1-4 alkyl, -CO-NH-C _1-4 alkyl, NO ₂ or NH ₂ ; stands
B represents phenyl, naphthalene, quinoline, isoquinoline, tetra hydroquinoline, tetrahydroisoquinoline, pyridine, pyrimidine, pyrazine, pyridazine, quinazoline, quinoxaline, thiophene, benzothiophene, benzofuran, furan or indole, the aromatic and heteroaromatic rings being represented by up to three radicals R ⁴ can be substituted,
in which
R ⁴ for C ₁ -C ₄ alkyl, -OC ₁ -C ₄ alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ -C ₄ -Alkyl, -NHCO-C ₁ -C ₄ -alkyl, -NHCOPh, -NHSO ₂ -C ₁ -C ₄ -alkyl, -NHSO ₂ -Ph, pyridine, -SO ₂ -C ₁ -C ₄ -alkyl or - SO ₂ Ph is;
X for a bond, - (CH ₂ ) _m -, - (CH ₂ ) _m -O- (CH ₂ ) _o -, -S- (CH ₂ ) _m , -SO- (CH ₂ ) _m -, -SO ₂ - (CH ₂ ) _m , -CH = CH-, -C5C-, -CO-CH = CH-, -CO- (CH ₂ ) _m -, - (CH ₂ ) _m -NHCO- (CH ₂ ) _o -, - (CH ₂ ) _m -CONH- (CH ₂ ) _o -, - (CH ₂ ) _m -NHSO ₂ - (CH ₂ ) _o -, - (CH ₂ ) _m -SO ₂ NH- (CH ₂ ) _o - or -NH-CO-CH = CH- stands
in which
m and o represent 0, 1, 2, 3 or 4;
R ^{2 represents} a C ₁ -C ₆ hydrocarbyl radical which can also carry a cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclo heptyl, indolyl, phenyl, pyridine or naphthyl ring, where these rings can in turn be substituted by one or two R ⁴ radicals as defined above, or for a —SCH ₃ radical; and
R ^{3 represents} hydrogen.

Group VIII according to DE-196 42 591

Compounds of formula I, wherein
R ¹ for

stands,
wherein
A for -CO-R ⁴ , -SO ² -R ⁴ , -CONH-R ⁴ , -COOR ⁴ , -C (= N) -R ⁴ , -C (= O) -NHR ⁴ or -C (= S ) -NHR ⁴ ; in which
R ^{4 represents} -C ₁ -C ₆ alkyl, where a chain of two or more carbon atoms can also contain a double bond or triple bond and / or with one or two substituents selected from phenyl, naphthalene, quinoxaline, quinoline, isoquinoline, Pyridine, thiophene, benzothiophene, benzofuran, pyrimidine, thiazole, isothiazole, triazole, imidazole, cyclohexyl, cyclopentyl, fluorene, indole, benz imidazole, oxazole, isooxazole and furan can be substituted, with each of the rings itself up to two radicals R ⁵ can carry
in which
R ⁵ for C ₁ -C ₄ alkyl, -OC ₁ -C ₄ alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ -C ₄ -Alkyl, -NHCO-C ₁ -C ₄ -alkyl, -NHCOPh, -NHSO ₂ -C ₁ -C ₄ -alkyl, NHSO ₂ -Ph, -SO ₂ -C ₁ -C ₄ -alkyl or -SO ₂ Ph stands;
R ^{2 represents} -C ₁ -C ₆ alkyl and can also carry a phenyl, pyridine or naphthyl ring which in turn can be substituted with up to two radicals R ⁵ as defined above; and
R ^{3 represents} -COOR ⁶ or -CONH-R ⁶ , where
R ^{6 represents} hydrogen, a phenyl ring which can carry one or two radicals R ⁵ as defined above, or represents C ₁ -C ₆ alkyl which can contain a double bond or a triple bond, and a ring such as phenyl, naphthalene, pyridine , Pyrimidine, piperidine, pyrrolidine, morpholine, thiophene, quinoline and isoquinoline, where the aromatic ring can also carry up to two residues -NR ⁷ R ⁸ or R ⁵ as defined above,
in which
R ⁷ and R ^{8 are} independently hydrogen or C ₁ -C ₆ alkyl.

Compounds of the group according to the invention are preferably used IX according to WO 99/54310

Compounds of formula I, wherein
R ¹ for

stands,
wherein
n represents 0, 1 or 2;
B represents phenyl, pyridine, pyridazine, pyrimidine or pyrazine;
R ⁴ for chlorine, bromine, fluorine, C ₁ -C ₆ alkyl, NHCO-C ₁ -C ₄ alkyl, NHSO ₂ -C ₁ -C ₄ alkyl, NO ₂ , -OC ₁ -C ₄ alkyl or NH ₂ stands,
D for a chemical bond, - (CH ₂ ) _z -O- (CH ₂ ) _z , (CH ₂ ) _m -, -CH = CH-
or -C∼C-, where
z represents 0, 1 or 2; and
m represents 0, 1, 2, 3 or 4;
A stands for - (CH ₂ ) _p -X
in which
p represents 1 or 2 and
X for pyrrolidine, morpholine, hexahydroazepine, piperidine, -NR ⁵ R ⁶ or

stands, wherein the cyclic amines can still be substituted with one or two radicals R ¹⁵ and R ^{15 represents} hydrogen, C ₁ -C ₆ alkyl, OC ₁ -C ₄ alkyl or phenyl; and
R ⁵ , R ⁶ and R ⁷ independently of one another represent hydrogen, C ₁ -C ₄ alkyl, cyclohexyl, cyclopentyl, CH ₂ Ph, Ph or CH ₂ CH ₂ Ph, it being possible for the phenyl rings to be substituted by R ⁶ ;
Y represents phenyl, pyridyl, pyrazyl, pyrimidyl or pyridazyl, it being possible for the rings to be substituted by up to two radicals R ⁸ , where
R ⁸ for hydrogen, C ₁ -C ₄ alkyl, -OC ₁ -C ₄ alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ - C ₄ alkyl, NHCO-C ₁ -C ₄ alkyl, -NHSO ₂ -C ₁ -C ₄ alkyl or -SO ₂ -C ₁ -C ₄ alkyl; or
A and Y together for

can stand where
R ^{16 represents} hydrogen, C ₁ -C ₆ -alkyl and (CH ₂ ) _b -phenyl with b = 1 to 4, it being possible for the phenyl ring to be substituted with up to two radicals R ⁶ as defined above,
R ^{2 represents} -C ₁ -C ₆ alkyl and can carry a SCH ₃ radical, a phenyl, imidazolyl, indolyl, cyclopentyl, cycloheptyl or cyclo hexyl ring, the ring having up to two radicals R ⁸ may be substituted as defined above; and
R ^{3 represents} hydrogen, COOR ⁹ or CO-Z, wherein
R ^{9 represents} hydrogen or C ₁ -C ₆ alkyl, which can be substituted by a phenyl ring, where the phenyl ring itself can also be substituted by one or two radicals R ¹² , where
R ¹² for hydrogen, C ₁ -C ₄ alkyl, -OC ₁ -C ₄ alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ - C ₄ alkyl, -NHCO-C ₁ -C ₄ alkyl, -NHCO-phenyl, -NHSO ₂ -C ₁ -C ₄ alkyl, -NHSO ₂ -phenyl, -SO ₂ -C ₁ -C ₄ alkyl or -SO ₂ phenyl; and
Z for NR ¹⁰ R ¹¹ ,

stands,
wherein
R ^{11 represents} hydrogen or C ₁ -C ₆ -alkyl, which can also be substituted by a phenyl ring, where the phenyl ring can also carry a radical R ⁹ as defined above, and
R ^{10 represents} hydrogen or C ₁ -C ₆ alkyl, which can be substituted by a phenyl ring, where the phenyl ring can also be substituted by one or two radicals R ¹² as defined above, or by

stands,
wherein
q represents 0, 1, 2, 3 or 4;
R ^{13 represents} hydrogen or C ₁ -C ₆ alkyl, which can be substituted by a phenyl ring, where the phenyl ring can also be substituted by one or two radicals R ¹² as defined above; and
R ^{14 represents} hydrogen or C ₁ -C ₆ alkyl, which can be substituted by a phenyl ring, where the phenyl ring can also be substituted by one or two radicals R ¹² as defined above.

In the above definitions of groups I to IX the given benen alkyl, alkenyl and alkynyl groups both straight-chain and also be branched.

For example, the following Group IX compound can be named:
N (1-carbamoyl-1-oxo-hexan-2-yl) -2 (E-2 (4 (N, N-dimethylaminomethyl) phenyl) ethen-1-yl) benzamide hydrochloride.

Among cysteine protease or calpain inhibitors for fiction According to its use, one also understands antisense molecules, ribozymes and other inhibitors that inhibit the expression of cysteine proteases or inhibit calpaines, such as specific ones Transkriptionssuppressoren. Also include inventive Inhibitors also antibodies and other molecules (as ge above called inhibitors) that bind to cysteine proteases or calpains and inhibit their activity. Include antisense molecules Nucleic acids attached to a cysteine protease or calpain code can bind rende mRNA and thereby the translation of the mRNA prevent. Ribozymes include enzymes that are selectively cysteine pro can hydrolytically cleave tease- or calpain-specific mRNAs NEN. In contrast, antibodies act directly on the cysteine protease or the calpain protein.

Corresponding protein or nucleic acid sequences are exemplary via sequence databases of the National Institute of Health (http: / / ncbi.nlm.nih.gov), the EMBL (http: / / www.embl.de), the DKFZ (http: / / www.dkfz-heidelberg.de) and the Sanger Institute (http: / / www.sanger.ac.nk) available.

According to the invention, the cysteine protease inhibitors can be used as such administered, but generally they are in the form pharmaceutical agents administered, d. H. as mixtures with we at least one suitable pharmaceutically acceptable carrier or diluent.

The cysteine protease inhibitors according to the invention can either as individual therapeutic agents or as mixtures with other therapeutic agents are administered, as in such active ingredients that the general constitution of the Improve patients, such as vitamins, antibiotics or anti-inflammatories. The cysteine protease inhibitors can on any suitable for the respective therapeutic purpose Way, e.g. B. orally or parenterally. The type of  pharmaceutical agent and the pharmaceutical carrier or of the diluent depends on the desired administration kind of.

Oral agents can be taken, for example, as tablets or capsules are present and can contain conventional excipients, such as bindemite tel (e.g. syrup, acacia, gelatin, sorbitol, tragacanth, or polyvi nylpyrrolidone), fillers (e.g. lactose, sugar, corn starch, Calcium phosphate, sorbitol or glycine), lubricants (e.g. Magnesi umstearate, talc, polyethylene glycol or silicon dioxide), desin tegrating agents (e.g. starch) or wetting agents (e.g. sodium lauryl sulfate). Antioxidants such as wise tocopherol flavor enhancing additives or stabilizers be included.

Oral liquid preparations or inhalants can be in the form aqueous or oily suspensions, solutions, emulsions, Siru pen, elixirs or sprays, etc. are present or can be as Troc ken powder for reconstitution in water or another egg suitable carrier. Such liquid preparations can be used Liche additives, such as suspending agents, flavor substances, diluents or emulsifiers.

Solutions or suspensions can be used for parenteral administration use with common pharmaceutical carriers. The par enteral application of cysteine protease inhibito according to the invention ren is advantageously carried out using a parenteral, in particular intravenously administrable, liquid pharmaceutical means. This preferably contains one for this Purpose suitable, pharmaceutically acceptable carrier, an effective same amount of at least one cysteine protease inhibitor, preferably wise in dissolved form. Examples of suitable pharmaceuticals shear carriers are in particular aqueous solutions, such as. B. physio logical saline solutions, phosphate-buffered saline, Ringer's solution, lactated Ringer's solution and the like. Moreover the agent can add other additives such as antioxidants, chelatbil contain agents or antimicrobial agents.

The pharmaceutical preparations according to the invention contain in addition the usual pharmaceutical excipients a therapeutically effective Amount of at least one cysteine protease inhibitor. Usually is this cysteine protease inhibitor in an amount of 0.001 to Contain 1 wt .-%, preferably 0.001 to 0.1 wt .-%.

The respective choice of the dosage of the cysteine according to the invention protease inhibitors and the respective dosing schedule are incumbent the decision of the attending doctor. This becomes dependent  on the chosen route of administration, on the effectiveness of the respective pharmaceutical agent, on the type and severity of the disease to be treated, the condition of the patient and his Response to therapy with a suitable dose and an ent Select the appropriate dosing schedule. So z. B. Invention appropriate cysteine protease inhibitors in a mammal (human and Animal) in doses of about 0.01 mg to about 100 mg per kg of body weight weight per day. You can in a single dose or administered in multiple doses.

The ent in addition to the cysteine protease inhibitor in the preparation substances and those in the manufacture of pharmaceutical The substances used in the preparations are toxicologically unsafe conceivable and ver with the respective cysteine protease inhibitor träglich. The pharmaceutical preparations are manufactured in the usual way, for example by mixing the cysteine pro tease inhibitor with other common carriers and diluents planning agents.

The invention will now be described with reference to the accompanying figures described in more detail using exemplary embodiments.

characters

Fig. 1 shows the weight of isolated perfused and ventilated rabbit lungs over time after the administration of activated granulocytes and perfusion with Krebs-Henseleit-Haes buffer (6.5% hydroxyethyl starch) containing 10 nM N (1-carbamoyl-1-oxo -hexan-2-yl) - 2 (E-2 (4 (N, N-dimethylaminomethyl) phenyl) ethen-1-yl) -benzamide hydrochloride or without calpain inhibitor.

example 1 Preparation of an isolated, perfused and ventilated Ka ninchenlunge 1. Experimental animals

20 rabbits were used as test animals for organ removal of such sex (bastards from Chinchilla and White New Zea land), all of which are owned by the Behring Werke Mar castle were delivered. The mean body weight of the rabbits was 3080 g.

2. Preparation of the experimental animals and model of the isolated, vent gelled and perfused rabbit lung

After inserting a vein cannula (scalp vein set) into the Ear veins of the experimental animal under local anesthesia were examined using this 1000 IU heparin / kg body weight (Liquemin, Hoffmann-La Roche AG, Gren zach-Wyhlen) administered.

The rabbit was then diluted with Nembutal 1: 2 (phenobarbital sodium, CEVA-GmbH, Bad Segeberg) narkoti (45-60 mg / kg body weight). After reaching one After anesthesia and relaxation, the animal was supine xiert and the local anesthesia for the tracheotomy performed. For this purpose 5-10 ml of xylocaine 2% below the larynx were placed in the injected subcutaneous adipose tissue (Astra Chemical GmbH, Wedel-Hol stone).

Then the preparation and attachment of the Trachea and the introduction of the plastic tube for ventilation of the Rabbit.

The lung was continuously ventilated with room air using a Starling pump (FA. Braun, Melsungen), taking 40 breaths / min. a tidal volume of 35 ml and a positive end-expiratory pressure (PEEP) of 1 cm H ₂ O was ventilated. The abdomen was then opened transversely with a costal margin incision, the diaphragm was detached ventrolaterally and the thorax was opened by a sternotomy.

After resection of the thymus and pericardium, the pulmonary artery was removed Close to the heart valve with a thread and an air-free, catheter filled with perfusate into the previously incised vessel introduced and fixed with thread.

The heart was down to the region of the exit of the pulmonary artery in the right ventricle and the junction of the pulmonary veins in the left atrium largely resected.

Then the trachea was dissected and cut through separation of the inferior esophagus and vena cava, the heart-lungs specimen was carefully removed from the thorax and together with the ventilation tubes and the pulmonary catheter on one egg electronic load cell (Hottinger Baldwin measurement technology) in attached to a humidified heating chamber (38 ° C) hanging freely. A minimal perfusion of 20 ml / min took place immediately after insertion the pulmonary catheter with a roller pump (Masterflex 756610, Cole Parmer Instrument Co., Chicago).  

A Krebs-Henseleit-Haes buffer (6.5% Hydroxyethyl starch) with an oncotic pressure of 25 mmHg, the was in two storage vessels heated to 38 ° C.

The 6.5% hydroxyethyl starch was made from HAES sterile (10%) From Fresenius AG, Bad Homburg.

Driven by the roller pump, the Perfusat came from the Storage vessels through a hose system via a Pall Ultipor Microfilter with 40 mm pore size (Pall Biomedizin GmbH, Dreieich) and a bladder trap in the pulmonary catheter.

The perfusion fluid emerging from the pulmonary veins was caught by the heat chamber and over a hose fed back to the storage vessels.

The circulation was started immediately after attaching the lungs to the load cell and the flow was gradually adjusted to 200 ml / min within 20 minutes. The pH value of the perfusion medium was regularly measured with an ABL2 (Radiometer, Copenhagen) and adjusted to 7.4 by adding CO ₂ to the ventilation air and kept constant. Without fumigation, the starting pH was 7.2-7.4.

The perfusate volume was 200 ml and was stored in the reservoir discarded and renewed until it is clear and bloodless was.

The weight of the isolated lungs was over the electronic weight cell detected, being set to 0 g at the start of the experiment and later the weight gain was evaluated in g. The pressure in the pulmonary artery was via a perfusion catheter lying and leading to the tip inner catheter by means of egg nes Statham Pressure Transducers added. Before the start of the experiment the pressure against atmospheric pressure was set to 0 mm Hg calibrates.

Registration of the two parameters, pulmonary arterial Pressure and weight, was carried out continuously with a ricadenki Multi-channel recorder (Rikadenki Electronics GmbH, Freiburg). It only such lungs were used for the experiments which, according to Er the maximum flow of 200 ml / min continuously over 30 Minutes a steady state in the behavior of the pulmonary artery pressure kes and the lung weight showed.

Example 2 Separation of human granulocytes

Density gradient centrifugation with polyvinylpyrrolidone-coated silicone particles (Percoll, Sigma Chemie GmbH, Munich, Order No. P 1644) is a quick and gentle method for isolating human PMN leukocytes from heparinized whole blood. It guarantees a 55% yield in 97% purity with full functionality of the isolated cells. For each isolation, 100 ml of heparinized venous whole blood from voluntary healthy donors were used, the mean being 215 × 10 ⁶ PMN. A 55% Percoll solution (D = 1.077 g / ml) and a 69% Percoll solution (D = 1.095 g / ml) were washed with distilled water. and freshly prepared 1.5 M NaCl daily. In each of 25 13 ml centrifuge tubes (Sarstadt, Nümbrecht), 4 ml of the 55% Percoll solution were placed first, then 4 ml of the 69% Percoll solution were carefully layered with a plastic spout. Then 4 ml of the heparinized whole blood were carefully overlaid.

After centrifuging the tubes at 350 g for 20 minutes (start-up time = 60 sec, braking time = 2 min) the plasma phase and the upper third of the 55% Percoll solution using a water jet pump sucked off. The underlying granulocyte band was carefully lifted off with a Pasteur pipette.

The cells were then washed by resuspension in Hanks-Hepes buffer and subsequent centrifugation. After aspiration of the supernatant, the cell pellet was resuspended in 50 ml of freshly prepared nutrient medium and for 90 minutes at 37 ° C. in a water bath in a gas atmosphere composed of 4% CO ₂ , 17% O ₂ and 79% N ₂ (Messer, Griesheim ) incubated at a gas flow of 2 l / min.

The cells were then washed again with H / H buffer and Centrifugation at 530 xg. After aspiration of the supernatant the pellet was filled with the test buffer (KHHB) and was able to can now be used in the experiment.

Example 3 Testing the effect of N (1-carbamoyl-1-oxo-hexan-2-yl) - 2 (E-2 (4 (N, N-dimethylaminomethyl) phenyl) ethene-1-yl) -benzamide Hydrochloride on that induced by granulocyte activation pulmonary edema

The benzamide was produced according to WO 99/54310 (Example 9 there) provides.

After a steady state of 20 minutes to obtain Aus output data were (at time 0) to induce a fault activated granulocytes according to pulmonary vascular permeability Example 2 in the pulmonary artery of a prepared according to Example 1 injected rabbit lung. For this, isolated human Granulocytes 10 minutes before their injection with heat denaturator activated immunoglobulin.

To test the inhibitor, it was perfused in one Concentration of 10 nmol / l before the steady state begins Phase added. Experiments without the addition of the Inhibitor.

In all experiments, an inhibitor cocktail (consisting of a cyclooxygenase blocker and a PAF, H ₁ and H ₂ antagonist) was added to the perfusate before the steady state phase began in order to influence the fluid balance by known vasoactive metabolites prevent.

Triggered by polytrauma, burns, intoxication and sepsis, shock and subsequently the development of an ARDS can occur, the symptoms of which are characterized, among other things, by the development of pulmonary edema. These pulmonary edema are mediated by activated granulocytes (PMN). In this way, isolated perfused and ventilated rabbit lungs after administration of humane PMN show a rapidly aggravating formation of edema that begins after 100 minutes, measured as an increase in lung weight. By applying 10 nmoles of N (1-carbamoyl-1-oxo-he xan-2-yl) -2 (E-2 (4 (N, N-dimethylaminomethyl) phenyl) ethen-1-yl) - benzamide beforehand On the other hand, hydrochloride can effectively prevent pulmonary edema (see FIG. 1).

Claims (6)

1. Use of at least one cysteine protease inhibitor for Manufacture of a pharmaceutical agent for the treatment of Diseases of the lungs. 2. Use according to claim 1, wherein the inhibitor is a calpain Is inhibitor. 3. Use according to claim 1 or 2, wherein the disease by Sepsis, skin burns, trauma and / or intoxication is triggered. 4. Use according to any one of the preceding claims, wherein it the lung disease is ARDS. 5. Use according to any one of the preceding claims, wherein the Inhibitor Capn1 (µ-Calpain) and / or Capn2 (m-Calpain) rever sibel inhibited. 6. Use according to any one of the preceding claims, wherein the inhibitor is selected from compounds of formula I.
wherein
R ¹ for
stands,
wherein
n represents 0, 1 or 2,
B represents phenyl, pyridine, pyridazine, pyrimidine or pyrazine;
R ⁴ for chlorine, bromine, fluorine, C ₁ -C ₆ alkyl, NHCO-C ₁ -C ₄ alkyl, NHSO ₂ -C ₁ -C ₄ alkyl, NO ₂ , -OC ₁ -C ₄ alkyl or NH ₂ stands,
D stands for a chemical bond, - (CH ₂ ) _z -O- (CH ₂ ) _z , - (CH ₂ ) _m -, -CH = CH- or -C∼C- and
z stands for 0, 1 or 2 and
m represents 0, 1, 2, 3 or 4;
A stands for - (CH ₂ ) _p -X
in which
p represents 1 or 2 and
X for pyrrolidine, morpholine, hexahydroazepine, piperidine, -NR ⁵ R ⁶ or
stands, wherein the cyclic amines can still be substituted with one or two radicals R ¹⁵ and R ^{15 is} hydrogen, C ₁ -C ₆ alkyl, OC ₁ -C ₄ alkyl or phenyl and
R ⁵ , R ⁶ and R ⁷ independently of one another are hydrogen, C ₁ -C ₄ alkyl, cyclohexyl, cyclopentyl, CH ₂ Ph, Ph or CH ₂ CH ₂ Ph, where the phenyl rings can also be substituted by R ⁶
Y represents phenyl, pyridyl, pyrazyl, pyrimidyl or pyridazyl, it being possible for the rings to be substituted by up to two radicals R ⁸ , where
R ⁵ for hydrogen, C ₁ -C ₄ alkyl, -OC ₁ -C ₄ alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ - C ₄ alkyl, NHCO-C ₁ -C ₄ alkyl, -NHSO ₂ -C ₁ -C ₄ alkyl or -SO ₂ -C ₁ -C ₄ alkyl;
or
A and Y together for
can stand where
R ^{16 represents} hydrogen, C ₁ -C ₆ -alkyl and (CH ₂ ) _b -phenyl with b = 1 to 4, it being possible for the phenyl ring to be substituted by up to two radicals R ^{6 in} accordance with the above definition,
R ^{2 stands} for -C ₁ -C ₆ alkyl and a SCH ₃ radical, a phenyl, imidazolyl, indolyl, cyclopentyl, cycloheptyl or cyclohexyl ring, the ring having up to two radicals R ⁸ may be substituted as defined above;
R ^{3 represents} hydrogen, COOR ⁹ or CO-Z, wherein
R ^{9 represents} hydrogen or C ₁ -C ₆ alkyl, which may be substituted by a phenyl ring, where the phenyl ring itself may also be substituted by one or two radicals R ¹² , where
R ¹² for hydrogen, C ₁ -C ₄ alkyl, -OC ₁ -C ₄ alkyl, OH, Cl, F, Br, J, CF ₃ , NO ₂ , NH ₂ , CN, COOH, COO-C ₁ - C ₄ alkyl, -NHCO-C ₁ -C ₄ alkyl, -NHCO-phenyl, -NHSO ₂ -C ₁ -C ₄ alkyl, -NHSO ₂ -phenyl, -SO ₂ -C ₁ -C ₄ alkyl or -SO ₂ phenyl; and
Z for NR ¹⁰ R ¹¹ ,
stands,
wherein
R ^{11 stands} for hydrogen or C ₁ -C ₆ alkyl, which may still be substituted with a phenyl ring, where in the phenyl ring a radical R ⁹ can still bear as defined above, and
R ^{10 represents} hydrogen or C ₁ -C ₆ alkyl, which can be substituted by a phenyl ring, where the phenyl ring can also be substituted by one or two radicals R ¹² as defined above, or by
stands,
wherein
q represents 0, 1, 2, 3 or 4 and
R ^{13 represents} hydrogen or C ₁ -C ₆ alkyl, which can be substituted by a phenyl ring, where the phenyl ring can also be substituted by one or two radicals R ¹² as defined above, and
R ^{14 represents} hydrogen or C ₁ -C ₆ alkyl, which can be substituted by a phenyl ring, where the phenyl ring can also be substituted by one or two radicals R ¹² as defined above; in which
in the above definitions, the alkyl, alkenyl and alkynyl groups given can be both straight-chain and branched.