CZ298197A3 - Reversible inhibitors of proteases - Google Patents

Abstract

The invention relates to novel reversible protease inhibitors. The inhibitors are specific to cysteine proteases. Examples of such inhibitors include compounds with structure (1).

Description

Reversible protease inhibitors

Technical field

The present invention relates to novel reversible protease inhibitors that are selective for cysteine proteases.

BACKGROUND OF THE INVENTION

Cysteine or thiol proteases contain cysteine in the active site for protolysis. Because cysteine proteases cause many diseases such as arthritis, muscular dystrophy, inflammation, cancer, glomerulonephritis, malaria and other parasite infections, their selective and irreversible inactivation provides opportunities for drug development. See, eg, Esser R. E. et al., Arthritis and Rheumatism (1994) 37, 236; Meijers, Μ. Η. M. et al., Agents Action (1993), 39, C219; Machleidt, W. et al., Fibrinolysis (1992), 6 Suppl. 4, 125; Sloan, B. F. et al., Biomed. Biochim. Acta (1991), 50,549; Duffy, M.J., Clin. Exp. Metastasis (1992), 10, 145; Rosenthal, P.J., Wollish, W.S., Palmer, J.T., Rasnick, D., J. Clin. Investigations (1991), 88, 1467; Baricos, W. H., et al., Arch. Biochem. Biophys. (1991) 288,468; Tomberry, N. A. et al., Nature (1992), 356,768.

Low molecular weight cysteine metalloprotease inhibitors have been described by Rich in Proteinase Inhibitors (Chapter 4, "Inhibitors of Cysteine Proteinases"), Elsevier Science Publishers (1986). Such inhibitors include peptide aldehydes that form hemithioacetals with a cysteine protease at the active site. See. e.g., Cheng, H., Keitz, P., and Jones, J. B., J. Org. Chem. (1994), 59, 7671. A disadvantage of aldehydes is their invivo and chemical instability.

Aldehydes were converted to α, β-unsaturated esters and sulfones by Wadworth-Emmons-Homer modification of the Wittig reaction (Wadworth, W. S., and Emmons, W. D., (J. Am. Chem. Soc. (1961), 83, 1733),

O

base _r ^ / EWG

where R = alkyl, aryl, etc., EWG = COOEt, SChMe, etc.

α, β-Unsaturated esters (Hanzlík et al., J. Med. Chem., 35 (6), 711-712 (1984); Tompson et al., J. Med. Chem., 29,104-111, (1986) , Liu et al., J. Med. Chem., 35 (6), 1067 (1992)) and α, β-unsaturated sulfones (Tompson et al., Supra, Liu et al., Supra) have been tested as inhibitors of two cysteine proteases, papain and dipeptidyl aminopeptidase I (also called cathepsin C). Inhibition of papain by these α, β-unsaturated compounds is poorly effective, as evidenced by a second order rate constant * that is less than 1 M ^ s ¹ resp. less than 70 m ^{_i _1} with the α, β-unsaturated esters and less than 20 ^M-1, respectively. less than 60 M < -1 & gt ^; for sulfones.

This has been further demonstrated on α-amino acid derivatives other than glycine or, in the case of an ester, on phenylalanine. Glycine derivatives are not chiral and phenylalanine derivatives are unclear, demonstrating that enzyme inhibition generally requires a chiral compound.

Some α-amino sulfonic acids have been proposed and prepared as potential inhibitory compounds, but their inhibitory effect has not been reported (McLainain et al., J. Chem. Soc. 75 (1941)).

It is further known that mannich condensation of sulfonic acid, aldehyde and ethyl carbamate produces urethanes (Engberts et al., Rec. 84: 942 (1965)).

Other procedures for selective and irreversible inhibition of cysteine proteases include alkylation with peptide α-fluoromethylketones (Rasnick, D., Anal. Biochem. (1985), 149, 416), diazomethylketones (Kirschke, H., Shaw, E., Biochem. Biophys Res. Commun. (1981), 101,454), alkoxymethylketones (Krantz, A. et al., Biochemistry, (1991), 30,

4678; Krantz, A. et al. No. 5,055,451, Oct. 8, 1991) and ketosulfonyl salts (Walker, B., Shaw, E., Fed. Proc. Fed. Am. Soc. Exp. Biol., (1985), 44, 1433).

Another class of cysteine protease inhibitors include epoxysuccinylpeptides containing E-64 and analogs thereof (Hanada, K., et al., Agric. Biol. Chem (1978), 42,523; Sumiya, S., et al., Chem. Pharm. Bull. (1992), 40,299; Gour Salin, BJ et al., J. Med. Chem., (1993), 36,720), α-dicarbonyl compounds reported by Mehdi, S., Bioorganic Chemistry, (1993), 21, 249 and N-peptidyl-O-acyl hydroxamates (Bromme, D., Neumann)

U., Kirschke, Η., Demuth, H-U., Biochim. Biophys. Acta, (1993), 1202, 271. For a further review of the procedures for reversible and irreversible inhibition of cysteine proteases see. Shaw, E., Advances in Enzymology and Related Areas of Molecular Biology (1990), 63,271.

SUMMARY OF THE INVENTION

The present invention provides protease inhibitors comprising targeting moieties linked by a chain of two carbon atoms with an electron-accepting group, wherein the dissociation constant for protease inhibition by this inhibitor (K i) is max. 100 μΜ.

Further, the present invention provides protease inhibitors comprising a targeting moiety linked either directly or through a carbon atom link or via a di-carbon chain to a sulfone group, wherein the dissociation constant for protease inhibition by this inhibitor (K i) is max. 100 µΜ.

Further, the present invention provides a compound which is a preferred protease inhibitor of Formula I:

Where:

n is 0 to 13;

AB is a linker selected from -C (O) NR ³ -, -CH ² NR ³ -, -C (O) CH 2 - and -NR ³ C (O) -, wherein R ³ is hydrogen or a group defined by m;

X is a bond, methylene, or -CH ₂ CH (R ⁴ ) -, wherein R ⁴ is hydrogen, alkyl or arylalkyl; Y is -CH (R ⁵ ) - or -NR ⁵ -, wherein R ⁵ is hydrogen or as defined below;

Z is - (CH ₂ ) ₂ -, -C (R ⁶ ) (R ⁷ ) - or -N (R ⁷ ) -, wherein R ⁶ is hydrogen or methyl and R ⁷ is defined as mze;

Z ¹ is - (CH ₂ ) ₂ -, -C (R ⁶ ) (R ⁸ ) - or -N (R ⁸ ) -, wherein R ⁶ is hydrogen or methyl and R ⁸ is defined below:

R ¹ is hydrogen, alkyloxycarbonylalkanoyl, alkyloxycarbonyl, alkanoyl unsubstituted or substituted with a carboxyl, alkyloxycarbonyl and heterocycloalkylalkanoylamino radical, cycloalkylcarbonyl, heterocycloalkylcarbonyl, heterocycloalkylcarbonyl, alkoxyalkylcarbonyl, alkoxyalkylcarbonyl, alkyl, alkoxycarbonyl, alkyl, alkoxycarbonyl, dialkylcarbamoyl, arylcarbamoyl, arylalkylcarbamoyl, arylalkanoyl, aryloyl, alkylsulfonyl, dialkylaminosulfonyl, arylsulfonyl or heteroarylsulfonyl;

R and R are independently hydrogen, alkyl unsubstituted or substituted by hydroxyl, amino, alkylamino, dialkylamino, uriedo, mercapto, alkylthio, carboxy, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, alkylsulfonyl and guanidine or protected derivatives thereof, cycloalkyl, cycloalkyl, cycloalkyl , heteroarylalkyl, an aryl and arylalkyl group unsubstituted or substituted on the aryl ring by one to three radicals selected from the group consisting of hydroxyl, amino, guanidino, halogen, optionally halogen-substituted alkyl, alkyloxy and aryl groups or protected derivatives thereof, or together with adjacent R ³ or R ⁵ groups form a divalent radical from the group consisting of (C 1-4) methylene and 1,2-phenylenedimethylene unsubstituted or substituted by hydroxy or protected by hydroxy or oxo;

and R is hydrogen, alkyl unsubstituted or substituted by one or more of amino, guanidino, halogen, hydroxy, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl or protected derivatives thereof, cycloalkyl, cycloalkylalkyl or aryl and arylalkyl groups unsubstituted or substituted on one aryl ring by one up to two radicals selected from the group consisting of amino, halogen, hydroxyl, optionally halogen-substituted alkyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl or protected derivatives thereof;

and pharmaceutically acceptable salts, individual isomers and mixtures of isomers thereof, wherein the dissociation constant of the protease inhibition by said inhibitor (K i) is preferably not more than 100 μΜ.

In addition, the present invention provides a compound that is a preferred protease inhibitor of Formula II:

R ^{1 / Y} 'Z'' ^A {b' ^xZ A / n

II where:

the groups have the same meaning as previously defined; and

R ⁹ is cyano, -C (O) 0R ^10, -P (O) (OR ¹⁰⁾ 2, -S (O) ^{(NR10) R10,} C (O) ^R, -S (O) ^R , -C (O) ^{NR 2} R ^13, -S (O) 2 ^{L 2} R ^13, -C (O) NHR ¹⁴ or -S (O) ₂ NHR ¹⁴ wherein R ¹⁰ is independently hydrogen, alkyl unsubstituted or substituted by one or more radicals from the group consisting of amino, halogen, hydroxyl, alkyloxy, nitro, alkylsulfonyl and ailylsulfonyl, or protected derivatives thereof, cycloalkyl, cycloalkylalkyl or aryl and arylalkyl groups optionally substituted on the aryl ring by one or two radicals from the amino group, halogen, hydroxyl, optionally halogen-substituted alkyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl, or protected derivatives thereof, R ¹¹ is hydrogen, alkyl, perfluoroalkyl, cycloalkyl, cycloalkylalkyl, perfluoraryl, perfluorarylaalkyl or aryl and arylalkyl groups optionally on the aryl ring substituted with one or two amino acid residues a halogen, hydroxy, optionally halogen-substituted alkyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl group, or protected derivatives thereof;

R ¹² and R ¹³ are independently hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl or arylalkyl, and R ¹⁴ is -C (O) OR ¹⁰ , wherein R ¹⁰ is as defined above or one of the substituents of formulas (a) and (b) :

wherein η, A, Β, Y, Z, R * and R ¹⁰ are as defined above, and their pharmaceutically acceptable salts, individual isomers and mixtures of isomers, wherein the dissociation constant of protease inhibition by said inhibitor (K i) is preferably max. 100 µΜ.

Further, the present invention provides a compound which is a preferred protease inhibitor of formula III:

ΠΙ in / ' ^A (b ^ ^Z ^ a

where:

the above groups are as defined above; and

R ¹⁵ is hydrogen, methyl, fluoro or groups of formula (a) and (b) as defined above, and R ¹⁶ is a substituent selected from the group consisting of phenyl or (C _5-6 ) heteroalyl, which is optionally substituted with at least one radical selected from the group containing alkylcarbamoyl, dialkylcarbamoyl, alkyloxycarbonyl, alkylsulfinamoyl, dialkylsulfinamoyl, alkylsulfonyl, carboxyl, nitro, sulfinamoyl, sulfo, carbamoyl, phosphono, alkyloxyphosphinyl, dialkyloxyphosphinyl, alkanoyl, cyanos, , optionally halo substituted alkyl, arylalkyl, halogen, - ⁺ N (R ¹⁷ ) 3, wherein R ¹⁷ is independently alkyl, aryl or arylalkyl, or -N (R ¹⁸ ) 2, wherein R ¹⁸ is independently hydrogen, alkyl, aryl or arylalkyl;

and pharmaceutically acceptable salts, single isomers and mixtures of isomers thereof, wherein the dissociation constant of the protease inhibition by said inhibitor (K i) is preferably max. 100 μΜ.

· · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a cysteine protease inhibitor of the present invention or one isomer, a mixture of isomers, or a pharmaceutically acceptable salt or salts thereof, in association with one or more pharmaceutically acceptable administration facilitators.

Further, the present invention provides a method for treating mammals in conditions that are ameliorated by inhibition of cysteine protease. The method comprises administering to said mammal a therapeutically effective amount of a cysteine protease inhibitor of the present invention or a single isomer, a mixture of isomers, or a pharmaceutically acceptable salt thereof.

Part of the present invention is a method for determining cysteine protease in a sample. This procedure includes;

(a) assaying said sample for protease activity using a protease substrate;

(b) assaying for protease activity in the presence of a known concentration of a cysteine protease inhibitor of the present invention; and (c) calculating the difference between a) and b) in detecting the cysteine protease-induced protease activity.

The present invention includes processes for preparing cysteine protease inhibitors of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 contains Scheme 1 - Synthesis of compounds of Formula I wherein X is a bond. The synthetic procedure is as follows: a) HCO2H, H2O; b) HBr / acetic acid; c) 4-methylmorpholine, isobutyl chloroformate, Mu-ROH; and d) chromatographic purification. The groups are defined in accordance with the present invention.

Figure 2 contains Scheme 2 - Synthesis of compounds of Formula I wherein X is a methylene group. The synthetic procedure is as follows: a) 4-methylmorpholine, isobutyl chloroformate, followed by reduction of NaBH ₄ in aqueous THF; b) CH ₃ SO ₂ Cl, triethylamine, ΟΙ ₂ Ο ₂ ;

c) R 15 SH, NaH, CH 3 OH, THF, heating; d) 4-chloroperbenzoic acid, CH 2 Cl 2;

θ ..... * ..... * ·· *

e) HCl / dioxane or p-CH 2 C 6 H 5 SO 5 / ether; and m) Mu-ROH, 4-methylmorpholine, isobutyl chloroformate.

Figure 3 contains Scheme 3 - the synthesis of compounds of Formula I wherein X is a methylene group. The synthetic procedure is as follows: a) (CH 3) ₃ CH ₂ CH ₂ SH, NaH, MeOH, THF, heating; b) 4-chloroperbenzoic acid, c) (nCi ₂ N + F, THF, followed by BrCH ₂ Cl, heating, d) HCl / dioxane or 4-CH ₃ C ₆ H ₄ SO ₃ H / ether; ae) 4-methylmorpholine, isobutyl chloroformate, Mu-PheOH.

Figure 4 contains Scheme 4 - synthesis of compounds of formula II. The synthetic procedure is as follows: ₂ N ⁺ (CH ₂ Cl ₂ , dicyclohexylcarbodiimide, Et 3 N / CH ₂ Cl ₂ ;

b) LiAlH / THF; c) NaH / THF; HCI / dioxane / CH ₂ Cl _2; e) 4-methylmorpholine, isobutyl chloroformate / THF; f) _H2, 5% Pd / C.

Figure 5 contains Scheme 5 - the synthesis of compounds of Formula I wherein X is ethylene. The synthetic procedure is as follows: a) (CH ₂ O) n, HCl, dioxane, eg where Ar = 2-naphthyl; b) (EtO) 3β;

c) CH ₃ CO 3 H, CH ₂ Cl ₂ ; d) NaH, THF; e) p-ClhCl / LH / ether; f) 4-methylmorpholine, isobutyl chloroformate; and g) H2, Pd / C.

Figure 6 describes the synthesis of compounds of formula II wherein R ⁹ is -COOH.

Figure 7 describes the synthesis of compounds of formula Π, wherein R ⁹ is -P (O) (R ¹⁰ ) ₂ . The synthetic procedure is as follows: a) NaH / THF; b) bezv. P-Cl2H2SO4H / ether;

c) 4-methylmorpholine, isobutyl chloroformate / THF, and; d) H ₂ , Pd / C.

Figure 8 describes the synthesis of compounds of formula II wherein R ⁹ is -C (O) NHR ¹⁴ . The synthetic procedure is as follows: a) NaOH / EtOH followed by HCl / H ₂ O; b) benzylamine, dicyclohexylcarbodiimide, CH ₂ C1 _2; c) NaH / THF, diethylbenzylamidomethylene phosphonate;

d) HCl / dioxane; e) 4-methylmorpholine, isobutyl chloroformate, THF; f) H ₂ , Pd / C, and as an alternative procedure for preparing from carboxylates, Scheme 6, supra; g) aniline, dicyclohexylcarbodiimide, CH ₂ C1 _2nd

Figure 9 describes the general synthesis of compounds of Formula II.

Figure 10 describes the general synthesis of compounds of formula ΠΙ. The synthetic procedure is as follows a) CH 3 CN or other suitable solvent, var. b) H ₂ O, NaOH, followed by extraction into an organic solvent; c) phosphorane, THF (Wittig reaction); d) P-CH 3 C 6 H 4 SO 3 H, ether; e) Mu-PheOH, 4-methylmorpholine, isobutyl chloroformate, THF; and 1) H ₂ , Pd / C.

The definition of "alkyl" in the terms alkyl, alkyloxy, alkylthio, alkylsulfonyl, alkylcarbamoyl, dialkylcarbamoyl, heteroarylalkyl, arylalkyl, and the like, means a straight or branched, saturated or unsaturated hydrocarbon radical having 1 to 10 carbon atoms (e.g. methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, vinyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methylallyl, ethynyl, 1-propynyl,

2-propynyl, etc.).

"Alkyloxyphosphinyl" and "dialkyloxyphosphinyl" mean the radicals -P (O) (OH) OR and -P (O) (OR) 2, respectively, where R is alkyl as defined above.

"Alkanoyl" in the words alkanoyl, alkanoyloxy, heterocycloalkylalkanoylamino and the like means a radical -C (O) R where R is an alkyl as defined above having a total of 1 to 11 carbon atoms or a designated number of carbon atoms - e.g. (C 1-6) alkanoyl refers to the formyl, acetyl, propionyl, isopropionyl, butyryl, isobutyryl, crotonoyl, isocrotonyl, etc. radicals.

"Aryl" means an aromatic monocyclic or polycyclic hydrocarbon radical having 6 to 14 carbon atoms or a designated number of carbon atoms and any cyclic ketone or thioketone derivative in which the free aromatic ring member is a member of the aromatic ring. Aryl includes, for example, phenyl, naphthyl, anthracenyl, phenanthrenyl,

1,2,3,4-tetrahydro-5-naphthyl, 1-oxo-1,2-dihydro-5-naphthyl, 1-thioxo-1,2-dihydro-5-naphthyl, etc.).

"Aryloyl" means a radical -C (O) Ar wherein Ar is aryl as defined above having a total of 7 to 15 carbon atoms or a designated number of carbon atoms; (C7-n) aryloyl is benzoyl, naphthoyl, etc.).

"Cycloalkyl", in the words cycloalkyl and cycloalkylalkyl, means a saturated or unsaturated, monocyclic or polycyclic hydrocarbon radical containing from 3 to 20 carbon atoms, or a designated number of carbon atoms, wherein the non-aromatic ring is a free bonded carbon atom and any cyclic ketone thereof and a thioketone derivative (e.g., the term cycloalkyl means cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, bicyclo [2,2,2] octyl, 1,2,3,4-tetrahydro-1-naphthyl, oxocyclohexyl, dioxocyclohexyl, thiocyclohexyl, 9 -fluorenyl, etc.).

"Halogen" means fluorine, chlorine, bromine or iodine.

"Heterocycloalkyl" in the words heterocycloalkyl, heterocycloalkylalkanoylamino, heterocycloalkylcarbonyl, heterocycloalkylcarbonyl, and the like means cycloalkyl as defined above, wherein 1 to 5 of the designated carbon atoms are replaced by the heteroaloma N, O, S, P or As, where the non-aromatic ring is and any heterocyclic ketone, thioketone, sulfone or sulfoxide derivative thereof (eg, the term heterocycloalkyl means piperidyl, pyrrolidinyl, pyrrolidinyl, imidazolidinyl, indolinyl, quinuclidinyl, morpholinyl, piperazinyl, N-methylpiperazinyl, piperidinyl, 4,4-dioxo-4- thiopiperidinyl, 1,2,3,4-tetrahydro-3-isoquinolyl, 2,4-diaza-3-oxo-7-thia-6-bicyclo [3.3.0] octyl, etc.). Hetero (C6) cycloalkyl includes morpholinyl, piperazinyl, piperidinyl and the like.

"Heteroaryl" means an aromatic monocyclic or polycyclic hydrocarbon radical containing a total of 5 to 14 atoms or a specified number of atoms, where 1 to 5 of the designated carbon atoms are replaced by a heteroatom selected from N, O, S, P or As, where the free bond atom is an aromatic ring and any heterocyclic ketone and thioketone derivative thereof (e.g., the term heteroaryl means thienyl, furyl, pyrrolyl, pyrimidinyl, isoxazolyl, oxaxolyl, indolyl, benzo [b] thienyl, isobenzofuranyl, purinyl, isoquinolyl, pterdinyl, pyrimidinyl, , pyridyl, pyrazolyl, pyrazinyl, 4-oxo-1,2-dihydro-1-naphthyl, 4-thioxo-1,2-dihydro-1-naphthyl, etc.). Thus, hetero (C6) aryl includes pyridyl, pyrimidinyl, and the like.

"1,2-Phenylenedimethylene" means a divalent radical of formula -CH 2 C 6 H 4 CH 2 -. For example, a group R'YZA- in which Y is -N (R ⁵ ), Z is -CH (R ⁷ ) -, A is carbonyl and R ⁷ together with R ⁵ forms 1,2-diphenylenedimethylene means a group of the following formula :

and substituted derivatives and individual stereoisomers and mixtures of stereoisomers. The substituted derivatives of the 1,2-phenylenedimethylene divalent radical may contain a hydroxy group on any carbon of the cyclic system or an oxo group on an unsaturated carbon cycle.

"Phosphono" means the radical -P (O) (OH) 2 "Methylene" as in "(C ^ jmetylen" and _"(C3 .7) methylene" means a straight, saturated divalent radical, who has the number of carbon atoms; (C 5-6 methylene) includes trimethylene (- (Ctb R 6) and tetramethylene (- (CH 2) 4 -). For example, a preferred embodiment of the present invention contains a proline residue as an ABZ group wherein AB represents CH 2 -NR ^{3 and} R ³ together with R ⁷ or R ⁸ forms and C 3 methylene. Thus, a group R 1 -YZA- in which Y is - (NR ⁵ ) -, Z is -CH (R ⁷ ) -, A is carbonyl and R ⁷ together with R ⁵ forms trimethylene means a group of the following formula:

and individual stereoisomers and mixtures of stereoisomers. Substituted derivatives of trimethylene and tetramethylene divalent radicals may contain hydroxy groups or protected derivatives thereof or an oxo group on any carbon cycle. Suitable hydroxy protecting groups are defined below.

"Oxa (C 3-7) methylene" and "aza (C 3-7) methylene" mean methylene as defined above, wherein of the designated carbon atoms it is replaced by an oxygen or nitrogen atom. E.g. "Oxa (C5) methylene" means 3-oxapentamethylene (-CH 2 CH 2 OCH 2 CH 2 -) and 2-oxapentamethylene (CH 2 OCH 2 CH 2 CH 2 -). Thus, -C (O) NR ²¹ R ²² represents a 4-morpholinylcarbonyl radical, wherein the radical is 4-morpholinylcarbonyl. ··· ··· ·· ·

R ²¹ and R ²² together form 3-oxapentamethylene and the remainder 1-piperazinylcarbanoyl, wherein R ²¹ and R ²² together form 3-azapentamethylene.

"Adjacent" used in the phrase "R ⁷ together with adjacent R ³ " means that the atoms to which R ⁷ and R ^{3 are} attached are alternately attached to each other.

'Animal includes humans, mammals (eg, dogs, cats, rabbits, cattle, horses, sheep, goats, pigs, game, etc.) and non-mammals (eg, birds, etc.).

"Disease" includes any pathological manifestations of animals and includes pathological manifestations that may be caused by the applied treatment, eg "side effects" of the treatment.

"Electronic Acceptance Group" (EWG) means a functional group that is generally a group capable of exerting a polarizing effect on the bond between itself and the carbon to which it is attached such that the electrons are polarized in favor of the electron attracting group. Polarizing properties are known to allow the electron attracting group to participate in hydrophobic interactions or hydrogen bonds with a cysteine protease active site to form enzyme inhibition. Generally, the electron acceptance group is preferred at the α-position of the phosphonium ylide of formula Ph3P = C (R) EWG, where it causes sufficient polarization to stabilize the ylide against the decomposition reaction with oxygen, water, hydrohalic acids, alcohols, etc. Preferred electron attracting groups are likewise, the ylides of formula (RO) 2P (O) C (R) EWG stabilize. These groups are cyano, -S (O) 2 R ² , -C (O) OR ¹⁰ , -P (O) (OR ¹⁰ ) 2-, -S (O) (NR ¹⁰ ) R ¹⁰ , C (O) R ^n, -S (O) ^R, -C (O) NR ¹² R ^13, -S (O) 2 NR ¹² R ^13, -C (O) NHR ^4, -S (O) 2 NHR ^'4, phenyl, and (C5-6) heteroaryl group, wherein R ^2, R ^{10, Rn, R12, R13} and ^R14 are as defined in the Summary of the invention. When the electron attracting group is phenyl or (C 5-6 heteroaryl), the ring may be substituted with one or more meta-conducting groups (eg, alkyloxycarbonyl, alkylsulfinamoyl, dialkylsulfinamoyl, alkylsulfonyl, carboxyl, nitro, sulfinamoyl, sulfo, phosphono, alkyloxyphosphinyl, alkyloxyphosphinyl, alkyloxyphosphinyl, alkyloxyphosphinyl, alkyloxyphosphinyl) , alkylsulfinyl, sulfamoyl, alkylsulfamoyl, dialkylsulfamoyl, alkyloxysulfonyl, disubstituted amino, trisubstituted ammonium, etc., ortho and para-conducting groups (e.g., hydroxy and alkyloxy, optionally halogen-substituted alkyl, aryl, arylalkyl, and the like) and electron-attracting groups (e.g., alkylcarbamoyl, dialkylcarbamoyl, alkyloxycarbonyl, alkylsulfinamoyl, dialkylsulfinamoyl, alkylsulfonyl, carboxyl, nitro, sulfinamoyl, sulfo, carbamoyl, phosphono, alkyloxyphosphinyl, alkyl, dialkyloxyphosphinyl, dialkyloxyphosphinyl, dialkyloxyphosphinyl, dialkyloxyphosphinyl, dialkyloxyphosphinyl, dialkyloxyphosphinyl) ulfamoyl, dialkylsulfamoyl, alkyloxysulfonyl, aryl, heteroaryl and the like).

The "leaving group" has the usual meaning used by organic chemistry. E.g. the atom or group capable of withdrawing under alkylation conditions is halogen and alkane- or arensulfonyloxy such as mesyloxy, ethanesulfonyloxy, benzenesulfonyloxy and tosyloxy and alkanesulfonylamino, alkanecarbonylamino, aminosulfonylamino, aminocarbonylamino and the like.

"Isomerism" is a phenomenon where compounds have the same summation formula but different nature or arrange the bonds of their atoms in space. Isomers that differ in the arrangement of atoms in space are called "stereoisomers". Stereoisomers that are not mirror images of each other are called "diastereoisomers" and stereoisomers that are unmistakable mirror images are called "enantiomers" or sometimes "optical isomers". A carbon atom bonded to four different substituents is called a "chiral center".

A compound with one chiral center has two enantiomers that have opposite chirality and their mixture is called a "racemic mixture". A compound having more than one chiral center has 2 ^η_1 enantiomeric pairs, where n is the number of chiral centers. Compounds with more than one chiral center may exist either as individual diastereomers or as a mixture of diastereomers and are termed "diastereomeric mixtures".

The compounds of formulas I, II and III may exist as individual stereoisomers or as mixtures of stereoisomers. For example, the compounds of formulas I, II and ΙΠ contain a chiral center on the carbon to which R ⁸ is attached. In addition, compounds of formulas I, II and III wherein Z is -C (R ⁶ ) (R ⁷ ) contain a chiral center on the carbon to which R ⁷ is attached. Thus, for example, compounds of formulas I, II and III wherein n is 0 and Z is -C (R ⁶ ) (R ⁷ ) will contain two chiral centers and will exist as four individual stereoisomers or mixtures thereof.

• • • •

“Absolute Configuration”. The individual stereoisomers are characterized by an absolute configuration at the chiral centers. The absolute configuration refers to the arrangement of substituents attached to the chiral center in space. Substituents attached to the chiral center are ranked according to the Cahn, Ingold, and Prelog sequence rules and then assigned the absolute configuration symbol R if the three highest priority substituents are arranged in space (with the fourth substituent having the lowest priority, facing away from the observer) from the highest-priority substituent to the lowest-priority substituent clockwise and the absolute configuration symbol S when the substituents are aligned counterclockwise. If one stereoisomer contains one chiral center, the absolute configuration describes the symbol R or S, followed by a dash and the chemical name of the compound. For the purposes of the present invention, if a single stereoisomer or mixture of stereoisomers contains two or more chiral centers, the compound is described by the absolute R or S configuration symbol immediately after the respective locant has been introduced. An acyl residue derived from a natural amino acid is described in the same way as the amino acid acyl residue that precedes the L symbol (eg, L-phenylalanine). Enantiomers of non-naturally occurring amino acid acyl residues are preceded by the symbol D. Preferably, the amino acid sides of the chains occur in (S) or L-forms, which is due to the stereospedity of the enzymes, although the D-forms may also be used in some cases. If no absolute configuration symbol is given at the ciliary center, it is meant that the description includes both configurations and mixtures thereof - racemic or otherwise. Thus, for example, a compound of the following formula:

has the title:

N ^2- (4-morpholinylcarbonyl) -N ^1- [3-phenyl-1S- (2-phenylsulfonylethyl) propyl] L-phenylalaninamide, wherein R ¹ is 4-morpholinylcarbonyl, R ⁸ is 2-phenylethyl and is on the same side reference areas as R ⁷ , R ⁷ is benzyl and R ¹⁹ is phenylsulfonyl;

O l

N -4-morpholinylcarbonyl-N- [3-phenyl-1- (2-phenylsulfonylethyl) propyl] -L-phenylalaninamide, wherein R ¹ is 4-morpholinylcarbonyl, R ⁸ is 2-phenylethyl and lies either on the same or on different sides reference areas, R ⁷ is benzyl and R ¹⁹ is phenylsulfonyl;

N ² -4-morpholinylcarbonyl-N- [3-phenyl-1S- (2-phenylsulfonylethyl) propyl] -β- (2-naphthyl) -Lalaninamide, wherein R ¹ is 4-morpholinylcarbonyl, R ⁸ is 2-phenylethyl and lies on the same side of the reference surface as R ⁷ , R ⁷ is 2-naphthylmethyl and R ¹⁹ is phenylsulfonyl and ethyl 4S- (N-4-morpholinylcarbonyl-L-phenylalanylamino) -6-phenylhexanoate, where R ¹ is

Λ

4-morpholinylcarbonyl, R is 2-phenylethyl and is on the same side of the reference surface as R ⁷ , R ⁷ is benzyl, and R ¹⁹ is ethoxycarbonyl.

In a preferred embodiment of the present invention, the compositions of the present invention are in the form of pure diastereomers. Alternatively, the compositions comprise mixtures of diastereoisomers. In a preferred embodiment of the present invention the content of one diastereomer is higher than 70% by weight, more preferably at least 90% by weight.

"Protecting group" has the usual meaning used in synthetic organic chemistry, ie a group that protects a reactive group in a molecule. See. See, e.g., Greene et al., Protective Groups in Organic Synthesis, Second Edition, John Wiley & Sons, 1991.

Examples of hydroxy protecting groups are heterocycloalkylcarbonyl such as

Examples of amino protecting groups include aryloxycarbonyl such as benzyloxycarbonyl and the like, aroyl such as benzoyl and the like, and oxycarbonyl such as ethoxycarbonyl and 9-fluorenylmethoxycarbonyl and the like. guanidino protecting groups include sulfonyl such as 2,3,5-trimethyl-4-methoxyphenylsulfonyl and the like. Examples of groups suitable for protecting the carboxy group by which ester groups are formed are C 4 -C 8 alkoxycarbonyl, preferably tert-butoxycarbonyl (BOC). ) or benzyloxycarbonyl (CBZ, Z), preferably cycloalkylaminocarbonyl or oxacycloalkylaminocarbonyl of 4 to 8 ring atoms, preferably 4-morpholinecarbonyl (Mu) and the like.

"Protected" in the description of a compound means a derivative of a compound or a group that has a blocked reactive site or sites via a protecting group.

·· φ φ ··

“Eventual” or “eventually” means a fact that may or may not occur, and the description includes instances where the fact occurs or is not. E.g. "Optionally further substituted with one or more functional groups" means that substituents may or may not be present in the disclosed compound and the present invention includes those compounds where one or more functional groups are present and compounds in which these functional groups are absent.

"Cysteine protease-associated diseases" in the present invention means pathological conditions associated with cysteine protease. Some diseases are associated with increasing levels of cysteine protease; such as arthritis, muscle distrophy, inflammation, cancer, and glomerulonitis. In other diseases, extracellular cysteine protease activity occurs which is not normally present in the tissues. In other cases, the cysteine protease is associated with the ability of germs such as viruses to infect or replicate in the host organism.

Specific examples of diseases associated with cysteine protease include, but are not limited to, arthritis, muscle distrophy, inflammation, cancer, glomerulonephritis, malaria, Alzheimer's disease, tumor metastasis, trauma, gingivitis, leishmaniasis, filariasis and other diseases of bacterial and parasitic origin. A typical example is the disease associated with the interleukin 1β (ICE) conversion enzyme.

"Pharmaceutically acceptable" means suitable for the preparation of a pharmaceutical composition which is generally safe, non-toxic and neither biologically nor otherwise undesirable and includes compositions suitable for veterinary use as well as for human pharmaceutical use.

"Pharmaceutically acceptable salt" means a salt that is pharmaceutically acceptable as defined above and that has the desired pharmacological activity. Such salts include salts of acids such as hydrochloric, hydrobromic, sulfuric, nitric, phosphoric and the like; or organic acids such as acetic, propionic, hexanoic, heptanoic, cyclopentanepropionic, glycolic, pyruvic, lactic, malonic, succinic, malonic, maleic, fumaric, tartaric, citric, benzoic, o- (4-hydroxybenzoyl) benzoic, cinnamon, methanesulfonic, ethanesulfonic, 1,2-ethanedisulfonic,

2-hydroxyethanesulfonic, benzenesulfonic, p-chlorobenzenesulfonic,

2-naphthalenesulfonic, p-toluenesulfonic, camphorsulfonic, 4-methylbicyclo [2.2.2] oct-2-en-1-carboxylic, glucohepton, 4,4'-methylene-bis (3-hydroxy-2-en-1-carboxylic),

3-phenylpropionic, trimethylacetic, tert-butylacetic, lauryl sulfuric, gluconic, glutamic, hydroxynaphthoic, salicylic, stearic, muconic and the like.

Pharmaceutically acceptable salts are also base salts formed by the reaction of an acidic proton with an inorganic or organic base. Suitable inorganic bases are, for example, sodium hydroxide, sodium carbonate, potassium hydroxide, aluminum hydroxide and calcium hydroxide. Suitable organic bases are ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine and the like.

"Therapeutically effective amount" means the amount which, when administered to animals for treatment:

(1) protects susceptible animals which have not yet undergone the disease or animals with symptoms of disease (2) attenuates the disease, or. halts its development (3) improves the condition of the disease, ie it removes the disease

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to novel cysteine protease inhibitors. Inhibitors are expected to bind to cysteine protease based on the following scheme.

A complex enzyme inhibitor

It is believed that the enzyme is thus reversibly inhibited by the interaction between the R, Y and Z groups of the inhibitor and the surface of the enzyme active site and by the hydrogen bond interaction between the sulfone group and the amino acid chain active site.

This mechanism of reversible inhibition allows the specificity of cysteine protease enzyme inhibitors. In general, the inhibitors of the present invention inhibit cysteine proteases and do not inhibit serine, aspartyl and zinc proteases. In some embodiments of the present invention, the protease inhibitors of the present invention may be active against other types of proteases such as serine, aspartyl or other metalloproteases, but only to a minor extent.

Furthermore, the electron acceptance properties of the sulfone group of the compound of formula I polarize electrons between the sulfone group and the carbon to which it is attached, thereby allowing hydrogen bonding between the compound of formula I and the active site of the cysteine protease, allowing close bonding between the inhibitor and cysteine protease. described. Obviously, further electron pull or electron polarization occurring between the sulfur atom and the oxygen atom is likely, allowing the oxygen atom to participate in the hydrogen bond with the active site of the remainder of the protease and thereby contribute to further inhibition of the enzyme.

The present invention generally provides novel peptide-based cysteine protease inhibitors for use as reversible cysteine protease inhibitors. By "cysteine protease inhibitor" is meant an inhibitor that inhibits cysteine protease. In a preferred embodiment of the present invention, the cysteine protease inhibitors are specific for cysteine proteases; that is, they do not inhibit other types of proteases such as serine, aspartyl or other metalloproteases. In an alternative embodiment, the cysteine protease inhibitors of the present invention may well inhibit other types of proteases.

By the term "reversible" is meant herein that the inhibitor binds non-covalently to the enzyme and thus differs from irreversible inhibition. See. Walsh, Enzymatic Reaction Mechanisms, Freeman & Co., N.Y., 1979. "Reversible" in this context is a term known to those skilled in the art. Furthermore, reversible inhibitors of cysteine protease are competitive inhibitors, i.e., they reversibly compete with the substrate in binding to the enzyme, wherein the binding of the inhibitor and the substrate is mutually advantageous. Further, the stoichiometry of inhibition is 1: 1, i.e., one inhibitor molecule inhibits one enzyme molecule.

·· ··· · · · ·······

9 · 9 9 9 9 9 9 9 9 9 9 9 9

-JQ 9 9 9 9 9 9 9

IV 9999 99 999

The cysteine protease inhibitors of the present invention are designed to bind reversibly to cysteine protease. This binding is accomplished by means of peptide-based structures that act as targeting that mimic naturally occurring substrates and / or inhibitors. "Peptide" for the purpose of the present invention means an amino acid or peptide-like structure wherein one or more peptide linkers (eg, -C (O) NR-) is replaced by a similar shape, eg, -CH ₂ NR-, C (O) CH 2 - or -NRC (O) and / or wherein unnatural amino acid substituents are present.

For the purposes of the present invention, a "targeting moiety" means a peptide residue of a cysteine protease inhibitor that allows the inhibitor to bind to a cysteine protease.

In a preferred embodiment of the present invention, the targeting group of a cysteine protease inhibitor comprises at least two amino acid side chains or side chain analogs linked via a peptide bond or a spatial analog. The target group may contain up to 15 amino acids or an analogue thereof, although inhibitors generally have 1 to 7 amino acids or an analogue, since smaller inhibitors are suitable for therapeutic use. Thus, in the compounds of formulas I, II and ΠΙ, n is preferably 0 to 13, more preferably 0 to 5 and most preferably 0 to 3.

As described for compounds of formulas I, II and III, the targeting moiety may be a naturally occurring or non-occurring peptide residue of the formula:

«! '' νΑ, 'Ύ < ⁸

Wherein R and R are naturally occurring or non-occurring amino acid analogs or substituents, which are described in more detail below. The inhibitor targeting moiety may also contain other R ¹ functional groups as described below.

The amino acid substituents of the targeting moiety are believed to interact with the surface of the active site of the protease and thus bind. Further, it is contemplated that the amino acid substituents approach the electron-attracting group (e.g., R ⁸ in the formula above), occupy S | the position of the active site of the substrate, and so the Pi is directed to the rest of the substrate. An inhibitor. Similarly, another adjacent amino acid substituent (eg, R ⁷ in the formula above) occupies the S2 position of the active site of the substrate, thus directing the P2 residue of the inhibitor. If present another amino acid substituents will occupy the S3, _S4 and further positions the substrate binding site and be designated as P3, P ₄ inhibitor. Another targeting moiety that can be attached to the electron-attracting moiety will include an amino acid substituent that occupies S1, S2 ', and other substrate binding site positions and will be designated as P3', P ₄ 'etc., the rest of the inhibitor.

Targeting groups for specific enzymes are labeled using rules that apply to substrate specificity in cysteine proteases (see "Proteinase Inhibitors", in Research Monographs in Cell and Tissue Physiology (1986), Barret et al., Volume 12, Chapter 4:

Inhibitors of Cysteine Proteinases, Daniel Rich, Elsevier, New York; and Tombeny et al.). For example, interleukin-I converts an aspartic acid-binding enzyme (ICE) (i.e.

2-carboxyethyl) in the P 1 position and the substituents alanine (methyl), valine (isopropyl) or histidine (4-imidazolylmethyl) in the P 2 position. Papain binds the substituents arginine, lysine, N-benzyloxycarbonyllysine (i.e., 4-benzyloxycarbonylaminobutyl), homophenylalanine (i.e., 2-phenylethyl), guanidinophenylamino (i.e., 4-guanidinobenzyl) or norluecin (i.e., butyl) substituents in the pi. tyrosine, fi- (2-naphthyl) alanine (i.e.,

2-naphthyl), leucine, norleucine, isolucine or alanine at the P2 position. Cathepsin B binds the substituents arginine, lysine, N-benzyloxycarbonyllysine, guanidinophenylalanine, homophenylalanine or norleucine in the Pj position and the phenylalanine, tyrosine,

3,5-diiodotrosine (i.e., 3,5-diiodo-4-hydroxybenzyl), fi- (2-naphthyl) alanine, arginine, guanidinophenylalanine or citrulline (i.e., 3-ureidopropyl) at the P2 position. Cathepsin L and cruzain bind the substituents arginine, lysine, homophenylalanine, guanindinophenylalanine, citrulline or norleucine in the Pi position and the phenylalanine, tyrosine or β (2-naphthyl) alanine substituents in the P2 position. Cathepsin S binds the substituents arginine, lysine, homophenylalanine, guanidinophenylalanine, citruilin or norleucine at the Pi position and the substituents phenylalanine, tyrosine, β- (2-naphthyl) alanine, valine, leucine, norleucine, isoleucine or alanine in the P2 position. DPP-1 binds the phenylalanine or tyrosine substituents at the P 1 position and no or alanine substituents at the P2 position. Calpain binds the substituents phenylalanine, tyrosine, methionine, β-methylsulfonylmethylalanine (i.

2-methylsulfonylethyl) or valine in the P 1 position and the substituents valine, leucine, norleucine or isoleucine in the P 2 position.

• ·

fi.

R and R are independently hydrogen, alkyl (optionally substituted with hydroxy, amino, alkylamino, dialkylamino, uriedo, mercapto, alkylthio, carboxy, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, alkylsulfonyl and guanidino, or a protected derivative thereof), cycloalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, ayl and arylalkyl substituent (which are optionally substituted in the aryl moiety with one to three residues including hydroxy, amino, guanidino, halo, optionally halogen-substituted alkyl, alkyloxy and aryl, or a protected derivative thereof) or together with adjacent R ³ or R ⁵ groups it forms one of the following divalent radicals: ((N-methylene and 1,2-phenylenedimethylene (which are optionally substituted with a hydroxy group, or a protected derivative thereof, or an oxo group)).

In accordance with the present invention, preferred R ⁷ and R ⁸ are side chain groups consisting of natural amino acids or homologous derivatives thereof. These groups include alanine (methyl), arginine (3-guanidinopropyl), asparagine (carbamoylmethyl), cituriline (3-ureidopropyl), aspartic acid (carboxymethyl), cysteine (mercaptomethyl), glutamic acid (2-carboxyethyl), glutamine (2). -carbamoylethyl), glycine (hydrogen), histidine (4-imidazolylmethyl), homophenylalanine (2-phenylethyl), homoserine (2-hydroxylethyl), isoleucine (1-methylpropyl), leucine (isobutyl), lysine (4aminobutyl), methionine (2 -methylthioethyl), β- (1-naphthyl) alanine (1-naphthylmethyl), β (2-naphthyl) alanine (2-naphthylmethyl), norleucine (butyl), norvaline (propyl), omitine (3-aminopropyl), phenylalanine ( benzyl), proline, sarcosine (methylaminomethyl), lilac (hydroxymethyl), threonine (1-hydroxyethyl), tryptophan (3-indolymethyl), tyrosine (4-hydroxybenzyl), and valine (isopropyl).

Preferred compounds in accordance with the present invention are set forth in the Summary of the Invention. Preferred compounds of formulas I, II and III include those wherein n is 0 to 5; AB is a linker of the formula -C (O) NR ³ -, wherein R ³ is hydrogen or may be defined; Y is -N (R ⁵ ) -, wherein R ⁵ is hydrogen or dininated below; Z is - (CH 2) 2 - or -C (R ⁶ ) (R ⁷ ) -;

Z is -CH (R) -; R is hydrogen, C 3 -C 10 alkyloxycarbonylalkanoyl, (C 1-9) alkoxycarbonyl, (C 2-10) alkanoyl (optionally substituted with one of the following: carboxy, (C 1-9) alkyloxycarbonyl, and hetero (C _4-8 ) cycloalkyl ( C2-io) alkanoylamino), (C ₄ .9) cycloalkylcarbonyl, hetero _(C4 .g) cycloalkylcarbonyl nesibstituovaný or substituted by one of the following radicals: hydroxyl, (C ₅₎ alkyl, (C ₅₎ alkanoyl, (Ci.sjalkyloxykarbonyl (C ₆ _io) aryl (C, 5) alkyloxycarbonyl and • ·

hetero (C4-8) cycloalkylcarbonyl, (C6-io) aryl (Ci. ₅₎ alkyloxycarbonyl, carbamoyl, (C | .5) alkylcarbamoyl, di (C-5) alkylcarbamoyl, (C6-io) arylcarbamoyl, (C6-io ) aryl (Ci_5) alkylcarbamoyl, (C ₆ .io) aryl (Ci_5) alkanoyl, (C ₇ -u) aroyl, (C-5) alkylsulfonyl, di (Ci_5) alkylaminosulfonyl, (C6-io) arylsulfonyl or hetero (C5 g) arylsulfonyl; and R and R are independently (C _1-5 ) alkyl unsubstituted or substituted with one of the following moieties: hydroxyl, amino, alkylamino, dialkylamino, uriedo, mercapto, alkylthio, carboxy, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, alkylsulfonyl and guanidino derivatives, or their protected, C 3-6 cycloalkyl, (C 3-7) cycloalkyl (C 1-5) alkyl, pyridyl, thienyl, furyl, imidazolyl, indolyl, pyridyl (C 1-4) alkyl, thienyl (C 1-4) alkyl, furyl (C 1-4) alkyl, imidazolyl ( C 1-6 alkyl, indolyl (C 1-6) alkyl, any of the following substituents: phenyl, naphthyl, phenyl (C 1-6) alkyl, naphthyl (C 1-6) alkyl, which are not or are not substituted in the aryl moiety by one to three of the following substituents: hydroxy, amino, chlorine, bromine, iodine, fluorine, methyl, trifluoromethyl, methoxy and phenyl or protected derivatives thereof, or together with adjacent R ³ or R ⁴ groups form one of the bivalent radicals including (C 3-6) methylene and 1,2-phenylenedimethylene which are not or are substituted by the following groups: hydroxy or protected derivatives thereof, or oxo.

In more preferred compounds of formula I, D and III, n is 0 to 2; AB is a linker of the formula -C (O) NR ³ wherein R ³ is hydrogen or may be defined; Y is -N (R ⁵ ) -, wherein R ⁵ is hydrogen or as defined below, Z is - (CH 2) 2 - or -C (R ⁶ ) (R ⁷ ), except when n is 0, Z is not ( CH2) _2- ; Z ¹ is -CH (R ⁸ ) -, R ¹ is hydrogen, (C 1-6 alkoxycarbonyl, (C 2-6) alkanoyl unsubstituted or substituted with any of the following radicals: carboxyl, (C 1-5) alkyloxycarbonyl and hetero (C4-8) cycloalkyl (C4 ^) alkanoylamino, -C (O) NR ^{2 R} R ²² wherein R ²¹ and R ²² together form aza (C24)) methylene, oxa _(C2 ^) methylene or _(C3 .7) methylene, (C4-8) cycloalkylcarbonyl , benzyloxycarbonyl, acetyl, benzyl or dimethylaminosulfonyl dimers;

and R ⁸ and R ⁷ are independently (C 5-6) cycloalkyl, (C 5-6) cycloalkylmethyl, 3-pyridyl, 2-thienyl,

2-furyl, 4-imidazolyl, 3-indolyl, 3-pyridylmethyl, 2-thienylmethyl,

2-furylmethyl, 4-imidazolylmethyl, 3-indolylmethyl, (C 1-6) alkyl unsubstituted or substituted by any of the following: mercapto, carboxy, amino, methylthio, methylsulfonyl, carbamoyl, dimethylcarbamoyl, guanidino and hydroxy;

..............

protected derivatives, phenyl, 1-naphthyl, 2-naphthyl, benzyl, 1-naphthylmethyl, 2-naphthylmethyl and

2-phenylethyl on the aromatic ring unsubstituted or substituted by the following radicals: hydroxy, amino, chlorine, bromine and fluorine or their protected radicals, or together with adjacent radicals R ³ or R ⁵ form one of the following bivalent radicals: (C 1-6 -methylene and 1, 2-phenylenedimethylene unsubstituted or substituted by hydroxy or its protected derivatives, or oxo.

Particularly preferred compounds of formulas I, II and III include those wherein n is 0 to 1, AB is one of the following linkages: -C (O) NR ³ -; Y is -N (R ⁵ ) -, wherein R ⁵ is hydrogen or as defined below; Z is -C (R ⁶ ) (R ⁷ ) -; Z 'is -GH (R ⁸ ) -; R ¹ is hydrogen, tert-butoxycarbonyl, benzyloxycarbonyl, acetyl, 3-carboxypropionyl,

3-methoxycarbonylpropionyl, biotinylaminohexanoyl, phenylacetyl, benzoyl, dimethylaminosulfonyl, benzylsulfonyl, 1-piperazinylcarbonyl, 4-methylpiperazin-1-ylcarbonyl or 4-morpholinylcarbonyl; R ⁷ is 3-pyridylmethyl, 2-thienylmethyl,

2-furylmethyl, 4-imidazolylmethyl, 3-indolylmethyl, (C 1-6) alkyl unsubstituted or substituted by one of the following moieties: mercapto, carboxy, amino, methylthio, methylsulfonyl, carbamoyl, dimethylcarbamoyl, guanidino and hydroxy, or protected derivatives thereof, benzyl , 1-naphthylmethyl, 2-naphthylmethyl and 2-phenylethyl on the aryl ring unsubstituted or substituted by one of the following groups: hydroxy, amino, chlorine, bromine and fluorine or their protected derivatives, or together with adjacent groups R ³ or R ⁵ form one of the following bivalent radicals: (C 1-4 -methylene and 1,2-phenylenedimethylene unsubstituted or substituted by hydroxy, or a protected derivative thereof, or oxo;

and R ⁸ is butyl, 2-phenylethyl, 2-methylsulfonylethyl, 2-tert-butoxycarbonylethyl,

2-tert-butoxycarbonylmethyl, 4-tert-butoxycarbonylaminobutyl, 4-benzoylaminobutyl or benzyloxymethyl.

Even more preferred compounds of formulas I, II and III are those wherein n is 0; AB is the following linker: -C (O) NH-; Y is -NH-; Z is -CH ^(R7) -; Z ¹ is -CH (R ⁸ ) -; R ¹ is hydrogen, tert-butoxycarbonyl, benzyloxycarbonyl, biotinylaminohexanoyl, benzoyl, piperizin-1-ylcarbonyl, 4-methylpiperazin-1-ylcarbonyl or 4-morpholinylcarbonyl;

R ⁷ is (C 1-5) alkyl, optionally substituted benzyl, 1-naphthylmethyl, 2-naphthylmethyl,

3-pyridinylmethyl or 2-methylsulfonylethyl groups; and R ⁸ is butyl, 2-phenylethyl or

2-methylsulfonylethyl.

The most preferred compounds of formula I, II and III are those wherein n is 0; AB is the following linker -C (O) NH-; Y is -NH-; Z is -CH ^(R7) -; Z ¹ is -CH (R ⁸ ) -; R ¹ is

1-piperisinylcarbonyl, 4-methyl-1-piperazinylcarbonyl or 4-morpholinylcarbonyl; R ⁷ is an unsubstituted or substituted benzyl, 1-naphthylmethyl or 2-naphthylmethyl group; and R ⁸ is 2-phenylethyl.

In general, preferred compounds of formula I include those wherein R ² is independently (C 1-6) alkyl unsubstituted or substituted with one or two of the following residues: amino, chloro, bromo, fluoro, hydroxy, and methoxy, or protected derivatives thereof, perhalogenated (C 1-6) -5) alkyl, (C 3-7) cycloalkyl, (C 3-7) cycloalkyl (C 1-5) alkyl or one of the following groups: phenyl, pentafluorophenyl, naphthyl and phenyl (C 1-6) alkyl unsubstituted or substituted on the aryl ring by one or two of the following residues: amino, chloro, bromo, fluoro, hydroxy, methoxy and halogen-unsubstituted or substituted methyl, or protected derivatives thereof, and R ⁴ is hydrogen, (C 1-5) alkyl or (C _6-10 ) aryl (C 1-5) alkyl.

More preferred compounds of formula I are those wherein R ² is (C 1-6) alkyl unsubstituted or substituted with one or two of the following residues: amino, chloro, bromo, fluoro, and hydroxy, or a protected derivative thereof, perfluorofC 1-6 alkyl, (C 5-6) (C _5-6 ) cycloalkylmethyl or one of the following: phenyl, naphthyl and benzyl unsubstituted or substituted by one of the following residues: aminohydroxy, chloro, bromo or fluoro, or protected derivatives thereof, and R ⁴ is hydrogen or methyl.

Particularly preferred are compounds of formula I wherein R ² is methyl, trifluoromethyl, and substituted or unsubstituted 2-naphthyl or 2-phenylethyl.

Most preferred compounds of formula I are compounds wherein R ² is phenyl,

2-naphthyl or 2-phenylethyl, preferably phenyl or 2-naphthyl, and R ⁴ is hydrogen.

Generally preferred are compounds of formula II wherein R ⁹ is -C (O) OR ¹⁰ , P (O) (OR ¹⁰ ) 2, -S (OXNR ^{, O} ) R 10, -C (O) NHC (O) R 10 or -S (O) 2 NHC (O) R ¹⁰ are compounds wherein R ^{10 is} independently (C 1-6) alkyl unsubstituted or substituted with one or two of the following groups: amino, chloro, bromo, fluoro, hydroxy and methoxy or protected derivatives thereof, (C 3-7) cycloalkyl, (C 3-7) cycloalkyl (C 1-5) alkyl, or one of the following: phenyl or phenyl (C 1-6) alkyl unsubstituted or substituted on the phenyl ring by one or two of the following substituents: amino, chloro, bromo, fluoro, hydroxy, methoxy and halogen unsubstituted or substituted substituted methyl, or protected derivatives thereof.

More preferred compounds of formula Π wherein R ⁹ is -C (O) OR ¹⁰ , -P (O) (OR ¹⁰ ) 2, S (O) (NR ¹⁰ ) R ¹⁰ , -C (O) NHC (O) R ^{, {>} or -S (O) 2 NHC (O) R ¹⁰ are compounds wherein R ¹⁰ is ethyl, (C 5-6) cycloalkyl, (C 5-6) cycloalkylmethyl or one of the following: phenyl and benzyl unsubstituted or substituted on phenyl of the ring with aminohydroxyl, chlorine, bromine or fluorine or protected derivatives thereof.

Generally preferred compounds of formula II wherein R ⁹ is C (O) ^R, or -S (O) R ^n, and R ¹¹ is (Ci.sjalkyl, (C 3-7) cycloalkyl, (C3-7) cycloalkyl ( C 1-5 alkyl or phenyl and phenyl (C 1-6) alkyl unsubstituted or substituted on the aryl ring with one or two of the following residues: amino, chloro, bromo, fluoro, hydroxy, methyl, trifluoromethyl and methoxy.

More preferred compounds of formula II in which R ⁿ C ^(O) R, or -S (O) ⁿ R are those compounds where R ^N is ethyl, cyclo (C _5-6) alkyl, cyclo (C5 ^) alkyl-methyl or phenyl and benzyl unsubstituted or substituted on the aryl ring by one or two of the following radicals: aminohydroxy, chlorine, bromine or fluorine or protected derivatives thereof.

Generally preferred are compounds of formula II wherein R ⁹ -C (O) NR ¹² R ¹³ or -S (O) 2 NR ¹² R ¹³ and R ¹² and R ¹³ are independently (C 1-5) alkyl, (C 3-7) cycloalkyl , (C 3-7) cycloalkyl (C 1-5) alkyl, or phenyl and phenyl (C 1-6) alkyl unsubstituted or substituted on the aryl ring with one or two of the following residues: amino, chloro, bromo, fluoro, hydroxy, methoxy and halogen unsubstituted or substituted methyl. More preferred compounds of formula (II) in which R ⁹ is R ⁹ .

...............

-C (O) NR ¹² R ¹³ or -S (O) 2 NR ¹² R ¹³ are compounds wherein R ¹² and R ¹³ are independently ethyl, (C 5-6 cycloalkyl, (C 5-6 cycloalkylmethyl or phenyl) and benzyl unsubstituted or substituted on aryl of the ring optionally substituted by one or two of the following residues: aminohydroxy, chloro, bromo or fluoro, or protected derivatives thereof.

Preferred compounds of formula Π in which R ^{9 is} -C (O) NHR ¹⁴ or -S (OHNHR ¹⁴ , wherein R ¹⁴ is a group of formula (a) and (b) are those wherein η, A, Β, Y Z, R ¹ and R ¹⁰ are as defined above with respect to preferred compounds of formulas I, II and III.

Generally preferred compounds of formula III are those wherein R ¹⁵ is one of the following groups: 2-furyl, 2-thienyl, 2-pyrrolyl, 2-phospholyl, 2-arsoyl, 3-pyridyl or

3-phosphinyl (which are optionally substituted with at least one of the following moieties: (C 1-5) alkylcarbamoyl, di (C _1-5 ) alkylcarbamoyl, (C 1-5) alkyloxycarbonyl, (C 1-5) alkylsulfinamoyl, di (C 1-5) alkylsulfinamoyl, ( C 1-6 alkylsulfonyl, carboxy, nitro, sulfinamoyl, sulfo, carbamoyl, phosphono, (C 1-5) alkyloxyphosphinyl, di (C 1-5) alkyloxyphosphinyl, (C _1-5 ) alkanoyl, cyano, (C 1-8) alkylsulfinyl, sulfamoyl, (C 1-6) alkylsulfamoyl, di (C 1-5) alkylsulfamoyl, (C 1-6) alkyloxy-sulfonyl, (C 1-5), phenyl, naphthyl, pyridyl, thienyl, furyl, imidazolyl, indolyl, hydroxy, (C _1-5 ) alkyloxy, optionally halogen substituted (C 1-8 alkyl, benzyl, halo, - ⁺ N (R ¹⁷ ) ₃ wherein R ¹⁷ is independently (C 1-8 alkyl, phenyl)

Or O (benzyl), or -N (R) 2, wherein R is independently hydrogen, (C 1-6) alkyl, phenyl or benzyl).

More preferred are compounds of formula III wherein R ¹⁶ is one of 2-furyl,

2-thienyl, 2-pyrrolyl, 2-phospholyl, 2-arsolyl, 3-pyridyl or 3-phosphinyl (optionally substituted with at least one of methylcarbamoyl, dimethylcarbamoyl, methyloxycarbonyl, methylsulfinamoyl, dimethylsulfinamoyl, methylsulfonyl, carboxy, nitro sulfinamoyl, sulfo, carbamoyl, phosphono, methyloxyphosphinyl, dimethyloxyphosphinyl, formyl, cyano, methylsulfinyl, sulfamoyl, methylsulfamoyl, dimethylsulfamoyl, methoxysulfonyl, methylsulfonimidoyl, phenyl, naphthyl, pyridyl, thienyl, methyl, indinyl, imidazolyl, imidazolyl, imidazolyl, imidazolyl, imidazolyl benzyl, halo, ⁺ N (R ¹⁷ ) 3, wherein R ¹⁷ is independently methyl, phenyl or benzyl, or -N (R ¹⁸ ) 2, wherein R ¹⁸ is independently hydrogen, methyl, phenyl or benzyl).

Generally preferred compounds of formula ΠΙ wherein R ¹⁶ is a group of formula (a) or (b) are those wherein n, A, Β, Y, Z, R ¹ and R ¹⁰ are as defined above with respect to preferred compounds of formula I, II and ΙΠ.

A preferred eysteine protease inhibitor in accordance with the present invention has an absolute configuration at all chiral centers (S). Preferred compounds of formula I wherein n is 0 but have the absolute configuration of the chiral center to which R ⁷ , (R) is attached. Preferred compounds of formula I are, for example, N ² - (4-morpholinylcarbonyl) -N ^1- (3-phenyl-1R-phenylsulfonylpropyl) -L-phenylalaninamide (compound 1), N ² - (4-morpholinylcarbonyl) -N ¹ - (3-phenyl-1S-phenylsulfonylpropyl) L-phenylaninamide (compound 2), N ^2- (4-morpholinylcarbonyl) -N '- (3-phenyl-phenylsulfonylpropyl) -L-phenylalaninamide (compound 3), N ² - ( 4-morpholinylcarbonyl) N '- (3-phenyl-1-benzylsulfonylpropyl) -L-leucinamide (compound 4),

N ² - (4-morpholinylcarbonyl) -N ¹ - (3-phenyl-1-trifluoromethylsulfonylpropyl) -L-phenylalaninamide (compound 5), N ² - (4-morpholinylcarbonyl) -N ¹ - (3-phenyl-1-benzylsulfonylpropyl) -1? -phenylalaninamide (compound 6), N ^2- (4-morpholinylcarbonyl) N ^, - (3-phenyl-1-phenylsulfonylpropyl) -L-leucinamide (compound 7),

N ² - (4-morpholinylcarbonyl) -N ¹ - (3-phenyl-1-fluoromethylsulfonylpropyl) -L-phenylalaninamide (compound 8), N ² - (4-morpholinylcarbonyl) -N ^, - (3-phenyl-1S-phenylsulfonylmethylpropyl) L-phenylalaninamide (compound 9); N- (4-morpholinylcarbonyl) -N- {3-phenyl-1S [2- (2-phenylethylsulfonyl) ethyl] propyl} -L-phenylalaninamide (Compound 10); N ^2- (4-morpholines (carbonyl) -N ^1- {3-phenyl-1 S- [2- (2-naphthylsulfonyl) ethyl] propyl) -β (2-naphthyl) -L-alaninamide (Compound 11); N-phenylacetyl-N ^1- [3-phenyl-1S (2-phenylsulfonylethyl) propyl] L-phenylalaninamide (compound 12), N - (N-benzyloxycarbonyl-β-alanyl) -N ¹ - [3-phenyl-1 S- ( 2-phenylsulfonylethyl) propyl] -L-phenylalaninamide (compound 13), 3- {2-phenyl-16- [3-phenyl-1S (2-phenylsulfonylethyl) propylcarbamoyl] ethylcarbamoyl) propionic acid (compound 14); 3- {2-naphthyl-11 E 3-phenyl-1 S- (2-phenylsulfonylethyl) propylcarbamoyl] ethylcarbamoyl) propionic acid (Compound 15); N ² - (4-morpholinylcarbonyl) N 1 - {3-phenyl-1 S- [2- (2-naphthylsulfonyl) ethyl] propyl} L-tyrosinamide (compound 16); methyl 3- {2-phenyl-1S- [3-phenyl-1S (2-phenylsulfonylethyl) propylcarbamoyl] ethylcarbamoyl) propionate (compound 17);

N ^2- (4-morpholinylcarbonyl) -N ^1- [3-phenyl-1 S- (2-phenylsulfonylethyl) propyl] -L-phenylalaninamide (Compound 18); N ² - (β-alanyl) -N '- [3-phenyl-1S (2-phenylsulfonylethyl) propyl] -L-phenylalaninamide (compound 19) and 5-phenylsulfonyl-3S- (N [N- (N-acetyl- L-tyrosyl) -L-Valyl] -L-alanylaminolvaleric acid (compound 20).

Preferred compounds of formula Π are: ethyl 4S- (N-benzylsulfonyl-p- (2-naphthyl) L-alanylamino) -6-phenylhexanoate (compound 21); elyl 4S- (N-benzylcarbamoyl-β (2-naphthyl) -L-alanylamino) -6-phenylhexanoate (compound 22); ethyl 4S- [N (4-morpholinylcarbonyl) -β-2- (naphthyl-L-alanylamino) -6-phenylhexanoate (compound 23); ethyl 4S- (N-benzylcarbamoyl-L-phenylalanylamino) -6-phenylhexanoate ( compound 24); ethyl 4S- [N- (4-morpholinylcarbonyl) -L-phenylalanylamino] -6-phenylhexanoate (compound 25); N ² - (4-morpholinylcarbonyl) -N '- [3-phenyl-1S- ( 2-phenylcarbamoylethyl) propyl] -L-phenylalaninamide (compound 26) and N- (4-morpholinylcarbonyl) -N- [3-phenyl-5- (2-benzylcarbamoylethyl) propyl] -L-phenylalaninamide (compound 27).

Preferred compounds of formula III are: N ^2- (4-morpholinylcarbonyl) -N ^1- {3-phenyl-S- [2- (4-methoxyphenyl) ethyl] propyl) -L-phenylalaninamide (compound 28) and N ² - (4 (morpholinylcarbonyl) -N ^1- {3-phenyl-1 S- [2- (4-aminophenyl) ethyl] propyl) phenylalaninamide (compound 29).

It will be apparent to those skilled in the art that Formula I includes the structure represented by the preferred Type IV below.

-WITH,

Ά '

IV

R * wherein M is 0,1 or 2 carbon atoms, AB is as defined above, R ¹ , R ² , R ⁷ and R ⁸ are as defined above, and Q is NH or 0 '. A preferred embodiment uses nitrogen in the AB junction at position B. In this embodiment, the number of carbon atoms between carbon to which it is about

attached by R, and the sulfur atom of the sulfone group defines whether the compound α, β, or γ-sulfone. As outlined in the Examples below, compounds can be termed aminosulfones using the amino acid name or chemical name.

• · · · · · · · · ·

Substance V is α-aminosulfone:

Vije β-aminosulfone:

r ’

Substance Vli is γ-aminosulfone:

Formula II includes a type VIII structure referred to as a γ-amino group, especially when R 9 is an electronceptive group:

In a preferred embodiment, the dissociation constant for the inhibition of protease by the inhibitor in accordance with the present invention is K i of 100 μΜ. The term "binding" or "dissociation" constant is identical and means an equilibrium dissociation constant for the reversible coupling of an inhibitor to an enzyme. The definition and the tentative dissociation constant are given below.

Determination of the dissociation constant is known in the art. For reversible inhibition reactions in accordance with the present invention, the reaction scheme is as follows:

Equation 3

TO

E + iy ^ · Ε k>

The enzyme and inhibitor are combined to form the E complex. This step is considered rapid and reversible and does not undergo any chemical changes. The enzyme and inhibitor are held together by non-covalent forces. In this reaction, k i is a second order rate constant and holds for the formation of the equilibrium complex EL. K k is a first order rate constant and holds for the dissociation of the equilibrium complex E L In this reaction, K i = k i / ki.

Measurement of the equilibrium constant Ki is known in the art and is described in the examples. In general, artificial colored and fluorescent substrates are used.

The corresponding K ₁ values can be read from the Dixon curve. See. Irwin Segel; Enzyme Kintics: Behavior and analysis of rapid eqinibrium and steady-state enzyme systems, 1975, Wiley-Interscience Publication, John Wiley & Sons, New York. An alternative is to calculate for competitive inhibition:

Equation 4

1- (v / v _o ) = (1) / ((1] + MI + QSJ / Km)))) where v _o is the rate of substrate hydrolysis in the absence of inhibitor and v; is the rate in the presence of a competitive inhibitor.

The dissociation constant is a very convenient method of quantifying enzyme activity on a particular substrate or inhibitor and is often used in the art. If the inhibitor shows a very low K i, it is a potent inhibitor. Therefore, cysteine protease inhibitors in accordance with the present invention have a dissociation constant Ki a maximum of 100 μΜ, preferably 10 μΜ, most preferably 1 μΜ.

CHEMISTRY • · · · · 1

The synthesis of inhibitors according to the present invention proceeds as follows. Compounds of formula I wherein X is a bond are prepared according to the procedure outlined in Scheme 1 in FIG. 1.

Treatment of a suitable aldehyde, such as isobutyraldehyde or hydrocinnamaldehyde, with a tert-butyl carbamate or benzylcarbamate, together with a sodium salt of a suitable sulfinic acid, such as benzenesulfinic acid (Aldrich Chemical Co.,) in the presence of aqueous formic acid provides the corresponding N-protected aminomethylsulfone. Benzyloxycarbonyl protected with aminomethylsulfones is deprotected with hydrogen bromide in acetic acid. Coupling with a suitable N-protected amino acid or peptide or peptide derivative provides a compound of formula I wherein X is a bond. Alternatively, a suitable N-terminal protected amino acid or peptide or peptide derivative such as N- (4-morpholinylcarbonyl) phenylalaninamide is reacted with a suitable aldehyde together with a sodium salt of a suitable sulfinic acid in the presence of aqueous formic acid to give a compound of formula I wherein X is binding.

Compounds of formula I wherein X is a methylene group are prepared according to the procedures in Schemes 2 and 3; 2 and 3.

Treatment of a suitable N-protected amino acid or peptide derivative with sodium borohydride provides the corresponding β-aminoethanol. Treatment of this alcohol with methanesulfonyl chloride in the presence of triethylamine gives the corresponding mesylate. Nucleophilic substitution with a thiol anion such as thiophenol, according to Spaltenstein, A., Carpion, P., Miyake, F., and Hopkings, P.B., J. Org. Chem. (1987) 52,3759, provides the corresponding β-aminosulfide. The sulfide is oxidized with 4-chloroperbenzoic acid to give the corresponding N-protected β-aminoethylsulfone. In a particular case, the mesylate is treated with a thiolate ion, such as the 2- (trimethylsilyl) ethanethiol derivative, the synthesis of which is described in Aerson, M. B., Ranasinghe, M. Palmer, J. T., Fuchs, P. L., J. Org. Chem. (1988) 53,3125, and then the corresponding 5-aminoethyl-2-trimethylsilylethylsulfide is obtained. The 2-trimethylsilylethylsulfide is reduced to the corresponding β-aminoethyl-2-trimethylsilylethylsulfone, which is then subjected to fluoride cleavage to remove trimethylsilyl fluoride and ethane as gaseous substances and the sulfinate intermediate which is alkylated in situ with a suitable halide derivative such as dry bromide derivative

...............

obtaining the corresponding N-protected O-aminoethyl halomethylsulfone. The N-protected O-aminoethyl halomethylsulfone sc takes place, and then fits with a suitable N-protected amino acid or peptide derivative to give a compound of formula I wherein X is methylene.

Compounds of formula (II) and (I) wherein X is an ethylene group are prepared according to the procedure outlined in Equations 5,6 and 7.

Equations 5

wherein a) is Cl-H ² N ⁺ (Me) OMe, dicyclohexylcarbodiimide, triethylamine; and

b) lithium aluminum hydride.

Equation 6 o

Equations 7

H

Boc-N.

EWG

HJPd

EW

R *

The appropriate N-tert-butoxycarbonylamino acid or peptide derivative is converted to the corresponding aminomethylaldehyde (e.g., see Fehrentz, J-A and Castro, B., Synthesis, (0), (1983), 676; Equation 5). The aldehyde is converted to the corresponding vinyl compound by Wittig or Wadsworth-Emmons-Homer modification of the reaction (e.g., see Wadsworth, J. Amer. Chem. Soc. 83: 1733 (1991), equation 6). The vinyl compound is reduced by catalytic hydrogenation (e.g., see Equation 7), and then deprotected and coupled with a suitable N-protected amino acid or peptide derivative to give the corresponding compound of Formula I or IL Alternatively, the vinyl compound is deprotected and coupled to the N-protected amino acid or peptide derivative to give the corresponding vinyl condensation product, which is then reduced to the corresponding compound of formula I or II.

Compounds of formula II are prepared according to Scheme 4, FIG. 4.

Conversion of the N-tert-butoxycarbonylamino acid or peptide dertivative to the corresponding aminomethylaldehyde is preferably carried out with N, O-dimethylhydroxylamine hydrochloride in the presence of triethylamine and dicyclohexylcarbodiimide in dichloromethane. Alternatively, the conversion is carried out with triethylamine and a benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate (BOP) condensation agent followed by reduction with lithium aluminum hydride to give the corresponding aldehyde (e.g., see Fehrentz, JA and Castro, B .; Synthesis, (1983)) , 676-678). The conversion of the aldehyde to the corresponding vinyl ester is carried out with sodium triethylphosphonoacetate. The vinyl ester is deprotected with hydrogen chloride in dioxane. Hydrogenation is typically carried out in the presence of palladium.

Compounds of formula I wherein X is an ethylene group are readily prepared by the procedure of Scheme 5, FIG. 5.

Treatment of the sodium salt of a suitable sulfonyl methanephosphonate (SMP) (e.g.

diethylphenylsulfonylmethanephosphonate, diethyl 2-naphthylsulfonylmethanephosphonate, diethylmethylsulfonylmethanephosphonate and the like) to the appropriate N-tert-butoxycarbonyl-aminoaldehyde prepared according to equation 5 provides the corresponding vinyl sulfone. The sulfone is deprotected with anhydrous p-toluenesulfonic acid in ether and then combined with the N-protected amino acid or peptide derivative to give the corresponding vinyl condensation product, which is then reduced to the corresponding compound.

4 4

4 4 4 44 ··· »

4 ·

4 4

444 4

4

The appropriate arylsulfonylmethanesophosphonates are prepared by treating the arylthiols with paraformaldehyde in the presence of hydrogen chloride and reacting with triethylphosphate to give the corresponding diethylphosphophonomethylarylsulfide, which is then oxidized. Alternatively, a suitable sulfide can be purchased (eg, diethylphosphonomethylmethylsulfide from Aldrich Chemical Co., diethylphosphonomethylphenylsulfide, etc.) and oxidized to the corresponding sulfone.

Compounds of formula (II) wherein R ⁹ is -COOH are prepared according to Scheme 6. 6.

The corresponding carboxylate is generally obtained by hydrolyzing a compound of formula Π wherein R ⁹ is -COOR ¹⁰ , followed by treatment with an acid to give the corresponding carboxylic acid.

Compounds of formula Π wherein R ⁹ is -P (O) (R ¹⁰ ) 2 are prepared according to Scheme 7 (Fig. 7) according to Scheme 6, but substituting the SMP with a sodium salt of a suitable methylenediphosphonate (eg, tetraethylmethylenediphosphonate, etc.) .

Compounds of formula II wherein R ⁹ is -C (O) NHR ¹⁴ are prepared according to Scheme 8 (Fig. 8) according to Scheme 6, but substituting SMP with a suitable diethylamido methylene phosphonate (eg, diethylbenzylamidomethylene phosphonate, etc.).

Suitable amidomethylene phosphonates are prepared by reacting the triethyl phosphonoacetate hydrolysis product with a suitable amine. Alternatively, compounds of formula II wherein R ⁹ is -C (O) NHR ^{14 are} prepared by reacting a compound of formula I wherein R ⁹ is COOH with a suitable amine. For example, the reaction is carried out in the presence of dicyclohexylcarbodiimide in CH 2 Cl 2 or by any other coupling method known in peptide chemistry.

In general, compounds of formula II are prepared according to Scheme 9 (Fig. 9), starting from compound I-VII.

···· · · · ···· «

Compound I

R ‘

The synthesis of ketones is carried out by the Wadsworth-Emmons reaction of the Boc-α-amino aldehydes with a suitable phosphonate and subsequent catalytic reduction with hydrogen on palladium. The aldehyde is generally prepared as described above. If the phosphonate is not commercially available, methyl or substituted methyl ketone, such as acetone or acetophenone, is prepared by treatment with diethyl chlorophosphonate. The enolate is generated, for example, by reacting a THF solution of diisopropylamine with butyllithium and then adding the ketone (to lithium diisopropylamide - LDA). See. ΗΌ. House, Modem Synthetic Reactions, 2nd Ed. (W. Benjamin, Inc., Menlo Park, CA, Chapter 9). After the enolate has been generated, diethyl chlorophosphonate is added. Combination of the enolate with diethyl chlorophosphate gives Wadsworth-Emmons reagent.

For the synthesis of cysteine protease inhibitors having nitrile as the EWG, compound II is used:

Compound Π

O

The synthesis of the nitriles is carried out by the Wadsworth-Emmons reaction of the Boc-α-amino aldehydes with suitable phosphonates and subsequent reduction in the presence of a suitable catalyst. The aldehyde is generally prepared as described above. The phosphonate is commercially available.

For the synthesis of cysteine protease inhibitors having sulfoxide as the EWG, compound III is used:

• · ···· · · · ··· · ··

Compound III

O O

Q «\ JI II ρ ς qk-c / ^x r '

The synthesis of the sulfoxides is carried out by the Wadsworth-Emmons reaction of the Boc-α-amino aldehydes with suitable phosphonates and subsequent reduction in the presence of a suitable catalyst. The aldehyde is generally prepared as described above. The phosphinate is prepared by treating the methyl sulfoxide salt with diethyl chlorophosphate. The salt is generated by adding BuLi to diisopropylamine followed by addition of methylsulfoxide.

For the synthesis of cysteine protease inhibitors having sulfonamide as the EWG, compound IV is used:

Compound IV

O oc, h _s o | J ||

C, H _S O || NHR ^h o

The synthesis of the sulfonamides is carried out by the Wadsworth-Emmons reaction of the Boc-α-amino aldehydes with suitable phosphonates and subsequent reduction in the presence of a suitable catalyst. The aldehyde is generally prepared as described above. The phosphonate is prepared e.g.

a) diethylphospholyl methanesulfonates prepared according to Carreter and Ghosez (Tetrahedron Lett., 28: 1104-1108 (1987)) are converted by treatment with phosphorus pentachloride to sulfonyl chlorides (M. Quaedvlieg, Metoden der Organische Chemie (Houben-Weyl), E. Muller, Thieme Verlag, Stuttgart, 4th Ed. 1955, Vol. IX, Chapter 14) or

b) treatment of the sulfonyl chloride with an amine such as ammonia, a primary amine (including amino acid derivatives) or a secondary amine, resulting in the formation of a sulfonamide (Quaedvlieg, supra, Chapter 19).

The sulfonamide phosphonate is then reacted with Boc-α-aminoaldehyde to give the target compound as in the Wadsworth-Emmons reaction.

For the synthesis of cysteine protease inhibitors having sulfinamide as the EWG, compound V is used:

Compound V

O O

QHPs.ll It

C 3 H 10 O ² NH ²

The synthesis of sulfinamides is carried out by the Wadsworth-Emmons reaction of Boc-α-amino aldehydes with suitable phosphonates and subsequent hydrogenation in the presence of a suitable catalyst. The aldehyde is generally prepared as described above. The phosphonate is prepared by one of the following methods. Treatment of thionyl chloride in the presence of aluminum chloride with methyldialkylphosphonates such as commercially available methyldiethylphosphonate (Aldrich) affords dialkylphosphorylmethanesulfinyl chloride (Vennstra et al., Synthesis (1975) 519 or Aerson, Comprehensive Organic Chemistry (Pergamon Press), Volume 3, chapter 11.18). . Alternatively, treatment of amines with dialkylphosphoryl sulfinyl chloride (Stirling, Internet. J. Sulfur Chem. (B) 6: 277 (1971)) affords dialkylphosphoryl sulfulinamide.

For the synthesis of cysteine protease inhibitors having sulfoximine as the EWG, compound VI is used:

Compound VI

CjHsO '

N.

^x r '°

The synthesis of the sulfoximines is carried out by the Wadsworth-Emmons reaction of the Boc-α-amino aldehydes with suitable phosphonates and subsequent hydrogenation in the presence of a suitable catalyst. The aldehyde is generally prepared as described above. The phosphonate is prepared in several ways. E.g. N-alkyl or N-arylphenylmethylsulfoximines are prepared according to Johnson, Comprehensive Organic Chemistry (Pergamon Press), chapter 11.11. Alternatively, the lithium salt of a compound such as N-alkylphenylmethylsulfoxitin (Cram et al., J. Amer. Chem. Soc., 92: 7369 (1970)) is prepared by treating the butyllithium in THF with a neutral substance. Reaction of this lithium salt with dialkyl chlorophosphates as commercially available diethyl chlorophosphate (Aldricb) affords the Wadsworth-Emmons reagent necessary for sulfoximine synthesis.

Compound VII is used for the synthesis of cysteine protease inhibitors having sulfonate as EWG:

Compound VII

O

The synthesis of the sulfonates is carried out by the Wadsworth-Emmons reaction of the Boc-α-amino aldehydes with suitable phosphonates such as diethylphosphoryl methanesulfonate and subsequent hydrogenation in the presence of a suitable catalyst such as Raney nickel. The phosphonate is prepared as follows. First, a salt of methyldialkylphosphonate such as commercially available methyldiethylphosphonate (Aldrich) is generated by treatment with a strong base such as LDA and sulfonated with sulfur trioxide / trimethylamine complex (Carreto et al., Tetrahedron Leti.,

28: 1104-1108 (1987)) to form diethylphospholyl methanesulfonate, which then reacts in the Wadsworth-Emmons reaction with an aldehyde to form α, β-unsaturated sulfonate.

The compound of formula II is prepared according to Scheme 10 (Figure 10).

The chloride containing R ⁸ and R ⁹ is generally prepared from commercially available Reagents and Substances by techniques well known in the art. The reaction generally provides a mixture of cis and trans isomers with a preference for the trans isomer. After reduction to a cysteine protease inhibitor in accordance with the present invention, the cis-trans isomerism in this embodiment disappears because only one substance is formed.

In one embodiment, the cysteine protease inhibitor of the present invention is further purified, if necessary, after synthesis, for example, from unreacted substances by, for example, crystallization or silica gel column chromatography eluting with solvent mixtures.

The following is a summary of methods for preparing compounds in accordance with the present invention:

(A) Preparation of a compound of formula TV:

wherein n is 0 to 12; R ²⁰ is cyano, -S (O) 2 R ² , -CH ² S (O) ₂ R ² , -CH ² CH (R ⁴ ) S (O) 2 R ² , - (CH 2) 2 C (O) OR ¹⁰ , - (CH 2) ₂ P (OXOR ^{, O} ) 2, - (CH 2) 2 S (O) (NR ¹⁰ ) R ¹⁰ , - (CH 2) ₂ C (O) R ^11! , - (CH2) 2S (O) R ^u, - (CH 2) ₂ C (O) NR ¹² R ^13, - (CH 2) 2 S (O) ² NR ² R ^i3, - (CH 2) ₂ C (O) NHR ¹⁴ , - (CH 2) 2 S (O) ² NH ¹⁴ or -CH ² CHR ¹⁵ R * ⁶ and A, Β, X, Y, Z, R ¹ , R ⁸ R ', R ² , R ¹⁰ , R ^h , R ¹² , R ¹³ , R ¹⁴ , R ¹⁵ and R ¹⁶ are as defined in the Summary of the Invention for compounds I, Π and ID and their pharmaceutically acceptable salts, individual isomers and mixtures thereof, by reaction of an amine of formula V:

R *

H, N

Kt with a compound of formula VI:

where _R , xK _z ^ ^A ^ B / ^z \ A ^ K _from Jk ₀₁₁ wherein n, A, Β, X, Y, Z, R ¹ , R ⁸ and R ²⁰ have been defined above;

(B) the preparation of a compound of formula IV wherein R 20 is -S (O) 2 R ² and its pharmaceutically acceptable salts, individual isomers and mixtures thereof is carried out by reacting a compound of formula VII:

R ¹

O

vil with an aldehyde of the formula R ⁸ CHO and a sodium sulphate of the formula R ² S (O) ONa, wherein n, A, Β, X, Y, Z, R 1 and R 8 are as defined above;

(C) the preparation of a compound of formula IV wherein R ²⁰ is -S (O) 2 R ² and its pharmaceutically acceptable salts, individual isomers and mixtures thereof is (1) reacting a compound of formula NH 2 P wherein P is a protecting group with an aldehyde of formula of formula R ⁸ CHO and sodium sulfinate of formula R ² S (O) ONa followed by deprotection to give a compound of formula VHI:

HjN SfOJjR ¹

VIII wherein R ² and R ⁸ are as defined in the Summary of the Invention for Compounds 1; and (2) reacting a compound of formula VIII with a compound of formula VI wherein n, A, Β, X, Y, Z and R ¹ are as defined above;

(D) the preparation of a compound of formula IV wherein R ²⁰ is -S (O) 2 R ² and its pharmaceutically acceptable salts, individual isomers and mixtures thereof is performed

(1) reaction of a compound of formula IX:

R ’

PHN

IX with a thiolate anion of formula RS ', wherein L is a leaving group and R and R ⁸ are as defined above to give a compound of formula X:

R *

SR *

PHN (2) oxidizing a compound of formula X to give a compound of formula XI:

R *

PHN

SiO ₂ R *

XI (3) reacting a compound of formula XI with a compound of formula VI, wherein η, A, Β, X, Y, Z and R * are as defined above;

(E) preparing a compound of formula IV, wherein R ²⁰ is cyano, -CH ² S (O) 2 R ² ,

- (CH 2) 2 C (O) OR 10, - (CH 2) 2 P (O) (OR 10) 2, - (CH 2) 2 S (O) (NR ^{, O} ) R ¹⁰ , - (CH 2) 2 C (O ) R ^u, - (CH2) 2S (O) R ", - (CH 2) ₂ C (O) NR ¹² R ^13, - (CH 2) 2 S (O) 2 NR ¹² R ^'3, - (CH2) 2C (O NHR ¹⁴ , - (CH 2) 2 S (O) 2NHR ^14, and pharmaceutically acceptable salts, individual isomers and mixtures thereof, is (1) reacted with a compound of Formula XII:

R *

PHN

CHO

With a compound of the formula selected from XIH and XTV:

fVO

R

XIII

XIV wherein R ⁸ and R ²⁰ are as defined above followed by deprotection to give a compound of formula XV:

XV (2) reacting a compound of formula XV with a compound of formula VI, wherein η, A, Β, X, Y, Z and R ¹ are as defined above and (3) reduction;

(F) the preparation of a compound of formula IV wherein R ²⁰ is -CH 2 CHR 4 R ¹⁶ and its pharmaceutically acceptable salts, individual isomers and mixtures thereof is (1) reacting a compound of formula XII with a compound of formula XVI

XVI wherein R ^8, R ¹⁵ and R ¹⁶ are as defined above, and then deprotecting to give a compound compound of formula XVII:

R ⁸ R ^{, p}

Nk

XVII (2) reacting a compound of formula XVII with a compound of formula VI, wherein n, A, Β, X, Y, Z and R? are defined as above and as defind above, and (3) by reduction;

(G) preferably further converting the free form (not a salt) of a compound of Formula IV to a pharmaceutically acceptable salt;

(H) preferred further conversion of salt IV to the free form; and (H) preferably further isolating the individual stereoisomers of the compound of Formula IV.

In one embodiment of the invention, the eystein protease inhibitor is labeled in accordance with the present invention. "Labeled" means that it has at least one element, an isotope, or chemical compound attached to detect an eystein protease inhibitor or its binding to an eystein protease. Generally, there are three types of marking:

(a) isotopes, which may be radioactive or heavy isotopes

(b) immuno-tagging, which may be an antibody or an antigen

(c) a coloring or fluorescent substance.

The marks may be in any position in the inhibitor, e.g. may be added as "R ^L" group in formula 1, or a radioisotope in any location. Examples of suitable labels are ⁴ C, ³ H and fluorescent labels, which are well known in the art.

PHARMACOLOGY AND USE

The prepared Eysteine protease inhibitors according to the present invention can be easily tested for their inhibitory effect. The inhibitor is first tested against the eystein protease, which is the intended inhibitor target, as mentioned above. An alternative is a series of cysteine proteases and their colored substrates, which are commercially available. Thus, various eysteine proteases are tested by synthetic techniques known in the art in the presence and absence of an eysteine protease inhibitor to confirm the inhibitory effect of the compound. Effective inhibitors are then subjected to kinetic analysis and the Ki value and dissociation constant are calculated.

• ·

When a compound inhibits at least one cysteine protease, it is considered to be a cysteine protease inhibitor for the purposes of the present invention.

In some cases, the cysteine protease is not commercially available in pure form and therefore the inhibitors of the present invention are tested biologically. For example, the inhibitor is added to cells or tissues that contain a cysteine protease and the biological effect is monitored.

In one embodiment, the inhibitor of the present invention is prepared or modified to reduce or stop protein degradation in vivo and in vitro. Generally, this is accomplished by incorporation into an artificial amino acid inhibitor, derivative or substituent. It is preferred that only one non-natural amino acid or amino acid side chain is introduced into the inhibitor, and thus the relationship between the inhibitor and the enzyme does not change significantly. However, some longer inhibitors that contain many residues that direct the inhibitor to the enzyme tolerate more than one synthetic derivative. The non-natural amino acid can be further modified to mimic the binding of the natural chain to the enzyme, which also leads to tolerance of more than one synthetic derivative. The degradation of the inhibitor also reduces or stops the use of a peptide of the same spatial structure, but of a different chemical composition.

In this embodiment, the resistance of the in vitro modified inhibitor to various known commercially available proteases is tested to determine their proteolytic stability. Promising candidates are then tested in a conventional manner in animal models, for example, using labeled inhibitors to determine their stability and efficacy in vivo.

Cysteine proteases that are inhibited by the compounds of the present invention have a thiol group at the active site. These proteases are found in bacteria, viruses, eukaryotic microorganisms, plants and animals. Cysteine proteases can generally be classified into one of four or more distinct classes. Examples of proteases inhibited by the substances according to the present invention are (but are not limited to) plant cysteine proteases papain, ficin, aleurain, oryzain and actinidain; animal cysteine proteases of cathepsins Β, H, J, L, N, S, T, O and C (cathepsin C is also known as dipeptidyl peptidase I), interleukin conversion enzyme (ICE), calcium-activated neutral proteases, calpain I and II; bleomycin hydrolase, picomian 2A and 3C viral cysteine proteases, aftovirus endopeptidase, cardiovirus endopeptidase, komvirus • endopeptidase, potyvirus endopeptidase I and Π, adenovirus endopeptidase, two endopeptidase cysteine and chestnut polysynthesis viruses, chestnut virus, and cysteine proteases known as essential for the life cycle of Plasmodia, Entamoeba, Onchocera, Trypansoma, Leishmania, Haemonchus, Dictyostelium, Therileria, and Schistosoma, as well as malaria (P. falciparium), trypanosomy (T cruzi, enzyme) proteases such as cruzain or cruzipain), murine P. vinckei, and C. elegans cysteine protease. For a detailed list of compounds inhibited by proteases in accordance with the present invention, see FIG. Rawlings et al., Biochem. J. 290: 205-218 (1993).

ADMINISTRATION AND PHARMACEUTICAL PRODUCTS

Cysteine protease inhibitors in accordance with the present invention are generally administered in a therapeutically effective amount by any suitable method known in the art, either alone or in combination with another cysteine protease inhibitor in accordance with the present invention or with another drug.

The therapeutically effective amount varies greatly depending on the degree of the disease, the age and condition of the patient, the efficacy of the compound used, and other factors.

A therapeutically effective amount of a cysteine protease inhibitor according to the present invention is 10 pg / kg body weight per day to 10 pg / kg body weight per day, typically 100 pg / kg / day to 1 mg / kg / day. Thus, a therapeutically effective amount in a human 80 kg human is 1 mg / day to 1000 mg / day, typically 10 mg / day to 100 mg / day.

The physician will be able to ascertain, without undue experimentation and personal experience and with this application, a therapeutically effective amount of a cysteine protease inhibitor in accordance with the present invention for a given disease.

In general, the cysteine protease inhibitors of the present invention are administered as a pharmaceutical composition by the following routes: orally, totally (e.g., through the skin, nose, lung or suppository) or outside the digestive system (e.g., intramuscularly, intravenously, lung, or under the skin) ). The composition is in the form of tablets, pills, capsules, partially solid material, powder, controlled release composition, solution, suspension, aerosol or any other suitable form and generally comprises cysteine protease inhibitors according to the invention.

with the present invention in combination with at least one pharmaceutically acceptable agent to facilitate administration, which is a non-toxic agent that does not adversely affect the therapeutic effect of a cysteine protease inhibitor in accordance with the present invention and can be any solid, liquid, partially solid or gaseous aerosol) means available to those skilled in the art.

The solid pharmaceutically acceptable administration agent is starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, milk powder and the like. the agent to facilitate administration is water, ethanol, glyeerol, propylene glycol and various oils such as petrolatum, animal, vegetable or artificial oils (eg peanut, soybean, mineral, sesame and the like oils). Preferred liquid carriers for injection are water, saline, aqueous dextrose and glycols.

Aerosol compositions utilize compressed gases. Inert gases suitable for this purpose are nitrogen, carbon dioxide, nitrous oxide and the like.

A.R. Alfonso Reminton's Pharmaceutical Sciences 1985, 17th ed. Easton, Pa .: Mack Publishing Company.

The amount of cysteine protease inhibitor in the composition varies considerably and depends on the type of composition, the size of the dosage unit, the administration aid used and other factors known to those of ordinary skill in pharmacology. Generally, the composition comprises 0.1 to 10 wt. % cysteine protease inhibitor, preferably 1 to 10 wt. The remainder of the composition are administration aids.

The preferred pharmaceutical composition is administered in unit doses for treatment or arbitrarily for symptom relief. A representative pharmaceutical composition comprising a cysteine protease inhibitor in accordance with the present invention is set forth in Example 20.

The following examples serve to describe in more detail the use of the subject matter of the invention and the best embodiment of the invention found. These examples are not intended to limit the scope of the invention but are merely illustrative. All references cited are intended to be deliberate references.

DETAILED DESCRIPTION OF THE INVENTION

For simplicity, the following abbreviations are used in the examples:

Mu = morpholine urea

Xaai = amino acid at position Pi relative to the active site of the enzyme Xaa ₂ = amino acid at position P ₂ relative to the active site of the enzyme γ-CO ₂ Et = γ-amino ethyl ester γ-SO ₂ F = γ-aminosulfone with phenyl terminator γ-CO ₂ H = γ-aminocarboxylate γ-PEt = γ-aminophosphonate γ-AM = γ-aminoamide γ-Ar (sub) = γ-aminoaromatic compound (suitably substituted) β-SO ₂ Ph = β-aminosulfone with phenyl substituent α-SO ₂ Ph = ? -aminosulfone with a phenyl substituent Hph = homophenylalanine

PSMP- diethylphenylsulfonylmethylene phosphonate

Np2 = 2-naphthylalanine SO ₂ 2NP = sulfone with 2-termination nafthylovým

Phac = phenylacetyl β-Ala = β-alanine

MeOSuc = methoxysuccinyl

E.g. Mu-Phe-Hph-D-SO ₂ F, wherein Xaa ₂ - Phe (phenylalanine) and Xaa ₁ - Hph (homophenylalanine), transformed into β-aminophenyl sulfone according to the procedure described in the Examples.

Example 1

Synthesis of a cysteine protease inhibitor containing γ-aminoester as EWG

Unless otherwise stated, all reactions were carried out under an inert atmosphere of argon or nitrogen at room temperature. THF was distilled from ketyl benzophenone and sodium. All other solvents and commercial chemicals were used without further purification.

···································

The synthesis of ethyl (S) -4- (4-morpholinecarbonyl-phenylalanyl) -amino-6-phenylhexanoate, abbreviated as Mu-Fe-Hph-γ-COjEt, was performed as follows:

Unless otherwise stated, all chemicals were purchased from Aldrich. 0.393 g 60 wt. sodium hydride dispersion in mineral oil (9.82 mmol) was added to a solution of triethylphosphonoacetate (2.20 g, 9.82 mmol) in THF (50 mL) at -100 ° C. The mixture was stirred for 15 minutes, and then a solution of Boc-homophenylalaninal (BocHphH) (2.35 g, 9.82 mmol) prepared by passing Boc-homophenylalanine (Synthetech) to its Ν, Ο-dimethylhydroxamide using the Fehrentz method was added. Lithium aluminum hydride (reduction) in THF (20 mL) was then added. The mixture was stirred for 45 minutes, and then 1 M HCl (30 mL) was added. The product was extracted with ethyl acetate (50 mL), washed with saturated NaHCO 3 (30 mL), dried over MgSO 4, filtered and evaporated to dryness. The dried residue was dissolved in CH 2 Cl 2 (10 mL), and a 4.0 M solution of HCl in dioxane (20 mL) was added. The mixture was stirred for 30 minutes, then the solvents were evaporated in vacuo and the remaining ethyl (S) -4-amino-6-phenyl-2-hexenoate hydrochloride was pump dried.

4-Morpholinecarbonylphenylalanine (Mu-PheOH, 2.74 g, 9.82 mmol, prepared according to Esser R., Arthritis & Rhumatism (1994), 37,236) was dissolved in THF (50 mL) at -100 ° C. Then 4-methylmorpholine (1.08 mL, 9.82 mmol) was added followed by isobutyl chloroformate (1.27 mL, 9.82 mmol). The mixed anhydride was stirred for 10 minutes, and then a solution of ethyl (S) -4-amino-6-phenyl-2-hexenoate hydrochloride from the previous step in DMF (10 mL) was added followed by 4-methylmorpholine (1.08 mL, 9.82 mmol). The mixture was stirred for 1 h, and then 1 M HCl (50 mL) was added. The product was extracted with ethyl acetate (100 mL), washed with saturated NaCl (50 mL), dried over MgSO 4 and decolourised with charcoal (DARCO), filtered and evaporated to dryness to give 3.80 g of intermediate (80% yield from Boc-). homophenylalaninal).

To a solution of this intermediate (1.45 g, 3.09 mmol) in ethanol (25 mL) was added 5 wt. palladium on carbon (0.5 g). The mixture was reduced in a Parr hydrogenator for 36 hours, then filtered and the solvent removed in vacuo to give 1.19 g (82%) of product.

All samples were subjected to thin layer chromatography (TLC) with UV indication at 254 nm followed by spraying with ninhydrin, bromocresol green or p49. · · · · · · · · · · · · · · · · · · · · · · · • Anisaldehyde. The retention factor (Rf) of Mu-Phe-Hph-γ-CO ₂ Et was 0.35 (5% MeOH / CH ₂ Cl ₂ ).

NMR spectra were measured on a Varian Gemini 300 MHz instrument. All 1H NMR data of this and the following examples are given as δ in ppm relative to tetramethylsilane and bold signal assignments. Explanation of abbreviations: s (singlet), d (doublet), t (triplet), q (quartet), br (broad). An asterisk (*) indicates that the signal is obscure or overlaps with another resonance.

Example 2

Synthesis of a cysteine protease inhibitor containing γ-aminosulfone as EWG.

Synthesis of (S) -3-tert-butoxycarbonylamino-5-phenyl-1-phenylsulfonylpentane (Boc-Hphy-SO ₂ Ph). To a solution of PSMP (8.87 g, 30.34 mmol) in THF (150 mL) at 0 ^Q C is added sodium hydride (1.21 g of 60 wt.% Dispersion in mineral oil). The mixture was stirred for 20 min, and then a solution of Boc-homophenylalaninal prepared according to Febrentz and Castro (7.99 g, 30.34 mmol) in THF (20 mL) was added. The solution was stirred at 0 ° C for 30 min, and then 1M HCl (100 mL) was added. The product was extracted into ethyl acetate (100 mL), washed with saturated NaHCO 3 (100 mL), saturated NaCl (50 mL), dried over MgSO 4, filtered, and the solvent removed in vacuo. The residue was dissolved in ethanol (100 mL) and transferred to a Parr bottle of 5 wt. palladium on carbon (0.92 g). The mixture was reduced in a Parr apparatus for 24 h. The solution was filtered through diatomaceous earth and the solvent was evaporated in vacuo. TLC of the product showed one product in quantitative yield, Rf 0.29 (30% v / v ethyl acetate / hexane), after spraying with paraanisaldehyde white color.

Example 3

Synthesis of a cysteine protease inhibitor containing γ-aminosulfone as EWG.

Synthesis of (S) -3-amino-5-phenyl-1-phenylsulfonylpentane hydrochloride (HCl Hph-γ-SO ₂ Ph).

To a solution of Boc-Hph-γ-SO ₂ Ph (12.24 g, 30.34 mmol) in dichloromethane (20 mL) was added hydrogen chloride in dioxane (50 mL of a 4.0 M solution). The mixture was stirred for 90 minutes, the solvent removed in vacuo and the residue dissolved in CH ₂ Cl ₂ (50 mL). The solution was •

carefully added to ether (500 mL) with stirring. The solid was filtered, washed with ether (50 mL) and dried in vacuo.

Example 4

Synthesis of a cysteine protease inhibitor containing γ-aminosulfone as EWG.

Synthesis of (S) -3- (4-morpholinecarbonylphenylalanyl) -amino-5-phenyl-1-phenylsulfonylpentane (MuPhe-Hph-y-SO ₂ Ph). To a solution of Mu-PheOH (2.94 g, 10.56 mmol) in THF (75 mL) at -10 ° C was added 4-methylmorpholine (1.16 mL, 10.56 mmol) and isobutyl chloroformate (1.37). mL, 10.56 mmol). The mixture was stirred for 5 minutes, and then (S) - (E) -3-amino-5-phenyl-1-phenylsulfonyl-1-pentene p-toluenesulfonate prepared by Wadsworth-Emmons condensation of Boc-homophenylalaninal and deprotected with p-toluenesulfonic acid (5) was added. 00 g, 10.56 mmol) and 4-methylmorpholine (1.16 mL, 10.56 mmol). The mixture was stirred for 45 minutes then diluted with ethyl acetate (100 mL), washed with 1 M HCl (2 x 50 mL), saturated NaHCO 3 (50 mL), saturated NaCl (50 mL), dried over MgSO 4, filtered and the solvent removed in vacuo. The residue was crystallized from CITCL / ether to give 4.27 g (72%) of the intermediate. 1.17 g of this material (2.08 mmol) was dissolved in ethanol (25 mL) and transferred to a Parr bottle with 5 wt. palladium on carbon (0.30 g). The mixture was hydrogenated overnight in a Pan apparatus at room temperature. Ethyl acetate was then added to the suspension of the product that crystallized from the reaction mixture. The solution was filtered and concentrated in vacuo, and the residue was crystallized from CH 2 Cl 2 / hexane. T _T = 176-178 ° C. TLC: (50% ethyl acetate / GLCl) Rf = 0.24.

Example 5

Synthesis of a cysteine protease inhibitor containing γ-aminosulfone as EWG.

Synthesis of (S) -3- (4-morpholinecarbonyltyrosyl) -amino-5-phenyl-1-phenylsulfonylpentane (MuTyrHph-γ-SChPh). To a solution of 4-morpholinecarbonyltyrosine (Mu-TyrOH prepared according to Esser, R., Arthritis & Rheumatism (1994), 37, 236, 0.50 g, 1.70 mmol) in THF (10 mL) at -10 ° C 4-methylmorpholine (0.187 mL, 1.70 mmol) and isobutyl chloroformate (0.22 mL, 1.70 mmol) were added. After 5 minutes HClHph-γ-SO 3 Ph (0.577 g, 1.70 mmol described in Example 3) and 4-methylmorpholine (0.187 mL, 1.70 mmol) were added. The mixture was stirred for 45 minutes and then ethyl acetate (50 mL) was added and the solution washed with 1M

HCl, saturated NaHCO 3 and NaCl (30 mL each), dried over MgSO ₄ , filtered, and the solvent was removed in vacuo. The residue was precipitated from CH ₂ Cl ₂ / ethylcr to give 0.58 g (59 wt%) of Mu-Tyr-Hph-γ-SO ₂ Ph. T _t 104-107 ° C, TLC (10 vol.% MeOH / CH ₂ C1 ₂₎ R _f = 0.59.

Example 6

Synthesis of a cysteine protease inhibitor containing γ-aminosulfone as EWG.

Synthesis of (S) -3- (4-2-morflinkarbonyl nafiyl-alanyl) amino-5-phenyl-l (2-naťitylsulfonyl) pentane (Mu-Np2-Hph-y-SO ₂ 2NP). 2-Naphthalethiol (9.64 g,

60.16 mmol) was dissolved in toluene (75 mL), and paraformaldehyde (3.97 g, 132 mmol) and HCl / dioxane (33 mL of 4.0 M solution) were added. The mixture was stirred for several days at room temperature, and then the solvent was evaporated in vacuo. The residue was suspended in hexane (200 mL), dried over MgSO ₄ , filtered and evaporated to dryness. The obtained crude chloromethyl-2-naphthylsulfide was combined with triethylphosphate (10.93 g, 65 mmol) and heated to boiling for 4 h. The mixture was then cooled to room temperature, diluted with ether (200 mL), washed with 1M HCl, saturated with NallCO 3, NaCl (150 mL each), dried over MgSO ₄ , filtered and concentrated in vacuo to give 17.35 g (93%) of diethyl 2-naphthyl thiomethylene phosphonate. This material was dissolved in CH ₂ Cl ₂ (300 mL) and cooled to 0 ° C. Peroxyacetic acid (23.5 ml of a 32% by weight solution in acetic acid (Aldrich Chemical Co.) was carefully added. The mixture was stirred overnight while warming to room temperature. The solution was washed with freshly prepared saturated aqueous sodium bisulfite solution (100 ml). ml), and then several times with saturated NaHCO 3 (until the aqueous phase was basic) The organic phases were dried over MgSO ₄ , filtered and the solvent removed in vacuo Chromatography on 60-200 mesh silica gel (0-10% v / v ethyl acetate / CH ₂ Cl ₂₎ ) afforded 6.5 g (34 wt.%) of the pure Wadsworth-Emmons reagent, diethyl 2-naphthylsulfonyl-methylene phosphonate together with the same amount of the mixture. TLC: (20 vol.% ethyl acetate / CH ₂ C1 ₂₎ R _f = 0.37 .

To a solution of diethyl 2-naphthylsulfonylmethylene phosphonate (3.91 g, 11.42 mmol) in THF (60 mL) at 0 ° C was added sodium hydride (0.457 g of a 60 wt% dispersion in mineral oil). The mixture was stirred for 15 minutes, and then Boc-homophenylalaninal (3.00 g,

11.42 mmol) in THF (5 mL). The mixture was stirred for 30 minutes, and then 1 M HCl · 4 was added

· · *:::.:.::..

(100 mL). The product was extracted with ethyl acetate (100 mL), washed with saturated NaHCO 3 (75 mL), saturated NaCl (50 mL), dried over MgSO 4, filtered, and the solvent removed in vacuo. The residue was dissolved in dichloromethane (10 mL), and HCl / dioxane (25 mL of a 4.0 M solution) was added to the solution. The mixture was stirred at room temperature for 1 h, poured into ether (300 mL) and filtered. The solid was washed with ether (2 x 50 mL) and dried in vacuo to give 3.30 g (74% w / w of Boc-homophenylalaninal) (S) - (E) -3-amino-5-phenyl-1- (2- naphthylsulfonyl) -1-pentene.

To a solution of Boc-2-naphthylalanine (2.68 g, 8.51 mmol, Synthetech, Ore) in THF (50 mL) at 10 ° C was added 4-methylmorpholine (0.936 mL, 8.51 mmol) and isobutyl chloroformate (1.103). mL, 8.51 mmol). The mixture was stirred for 5 minutes, and then (S) - (E) -3-amino-5-phenyl-1- (2-naphthylsulfonyl) -1-pentene (3.30 g, 8.51 mmol) and 4-methylmorpholine (0.936) were added. mL, 8.51 mmol). The mixture was stirred for 45 minutes, diluted with ethyl acetate (100 mL), washed with 1M HCl (50 mL), saturated NaHCO 3 (50 mL) and NaCl (50 mL), dried over MgSO 4, filtered and the solvent removed in vacuo. The obtained intermediate (S) - (E) 3- (tert-butoxycarbonyl-2-naphthylalanyl) amino-5-phenyl-1- (2-naphthylsulfonyl) -1-pentenbyl was crystallized from C1 hCL / ether / hexane in 69% yield. wt. The obtained material was dissolved (3.83 g, 5.90 mmol) in CH 2 Cl 2 (5 mL) and HCl / dioxane (15 mL of a 4.0 M solution) was added. The mixture was stirred at room temperature for 1 h, then poured into ether (500 mL) with stirring and filtered. The solid was washed with ether (2 x 50 mL) and dried in vacuo to give (S) - (E) -3- (2-naphthylalanyl) -amino-5-phenyl- (2-naphthylsulfonyl) -1-pentene (3, 5). 41g, 99% yield).

This material (2.00 g, 3.42 mmol) was dissolved in THF (15 mL) and cooled to 0 ° C. Then 4-methylmorpholine-carbonyl chloride (0.40 mL, 3.42 mmol) and triethylamine (0.953 mmol) were added. The mixture was stirred at 0 ° C for 1 h and then at room temperature for 2 h. Ethyl acetate (50 mL) was then added and the solution washed with 1M HCl (30 mL), saturated NaHCO 3 (30 mL), NaCl (30 mL), dried over MgSO 4, filtered and evaporated to dryness to give 1.58 g (69% w / w). of (S) - (E) -3- (4-morpholinecarbonyl-2-naphthylalanyl) amino-5-phenyl-1- (2-naphthylsulfonyl) -1-pentene. TLC: (50% ethyl acetate / hexane) R _f = 0.37.

This material (0.73 g, 1.10 mmol) was dissolved in ethanol (20 mL) and transferred to a Parr bottle with 5 wt. palladium on carbon (0.30 g). The mixture was reduced in a Parr apparatus for 36 h. The solution was then filtered and the solvent removed in vacuo.

·· ·· »· κο · · · · · · · ·

The product was purified by column chromatography on 60-200 mesh silica gel (50% v / v ethyl acetate / ethyl acetate as eluent) to give give 0.14g (19% w / w) of pure Mu-Np2-Hph-γ-SO22Np product along with the mixture. TLC: (50% ethyl acetate / CPLCL) R f = 0.34.

Example 7

Synthesis of a cysteine protease inhibitor containing γ-aminosulfone as EWG.

Synthesis of 3-acetyltyrosylvalylalanylamino-4-hydroxy-carbonyl-1-phenylsulfonylbutane (Ac-Tyr-Val-Ala-Asp-γ-SOiPh). To a solution of diethylphenylsulfonylmethylene phosphonate (3.58 g, 12.23 mmol) in 50 mL of THF was added sodium hydride (0.489 g of a 60 wt% dispersion in mineral oil, 12.23 mmol) at 0 ° C. The mixture was stirred for 15 minutes, and then a solution of Boc-AspH (PO-tBu) (3.04 g, 11.12 mmol) (prepared by converting Boc-Asp (PO-tBu) to its Ν, Ο-dimethyldroxamide and reduction) was added. lithium aluminum hydride) in THF (10 mL). The mixture was stirred for 1 h and then 1M HCl (30 mL) was added. The product was extracted with ethyl acetate (100 mL), washed with saturated NaHCO 3 (30 mL), NaCl (30 mL), dried over MgSO 4, filtered and evaporated to dryness to afford the intermediate. Silica gel chromatography (20-30% ethyl acetate / hexane, gradient elution) afforded 2.07 g, 45 wt. of intermediate (S) - (E) -3-tert-butoxycarbonylamino-4-tert-butoxycarbonyl-1-phenylsulfonyl-1-butene. This material was dissolved in ether (2 mL) and a solution of anhydrous p-toluenesulfonic acid (1.0 g, 5.87 mmol) in ether (2 mL) was added. The mixture was stirred at room temperature overnight, then diluted with ether (25 mL) and the white precipitate filtered, washed with ether and dried in vacuo to give 0.80 g (95%) of additional intermediate (S) - (E) -3. amino-4-tert-butoxycarbonyl-1-phenylsulfonyl-1-butene-p-toluenesulfonate.

This compound was coupled using mixed anhydride chemistry methods with Ac-Tyr-Val-AlaOH prepared by conventional peptide chemistry methods to give another intermediate (S) - (E) -3-acetyltyrosylvalylalanylamino-4-tert-butoxycarbonyl-1-phenylsulfonyl-1 -butenu.

This was treated with trifluoroacetic acid to remove the aspartic acid t-butyl ester in the side chain to give (E) -3-acetyltyrosylvalylalanylamino-4-hydroxycarbonyl-1-phenylsulfonyl-1-butene. This material (0.28 g, 0.444 mmol) was dissolved in ethanol (10 mL) and transferred to a Parr bottle with 5 wt. palladium on carbon (0.1 g). The solution was reduced overnight in a Parr apparatus, then filtered, and filtered.

the solvent was removed in vacuo. The residue was dissolved in methanol (5 mL) and diluted 40x with 1: 1 CH 2 Cl 2 / ether gave a gelatinous precipitate which, after separation on a Buchner funnel, yielded 0.18 g (64%) yield. The ratio of the S and R isomers to the Asp residue was 3: 1 (according to the doublet integral of the aromatic NMR region belonging to the Tyr residue).

Example8

Synthesis of a cysteine protease inhibitor containing γ-aminocarboxylate as EWG.

Synthesis of (S) -4- (4-morpholinecarbonylphenylalanyl) amino-6-phenylhexanoic acid MuPhe-Hph-γ-CChH. To a solution of Mu-Phe-Hph-γ-CChEt prepared according to Example 1 (0.5g, 1.06 mmol) was added aqueous NaOH (1 mL of a 2M solution). After 4 h the reaction was complete. 1M HCl (4ml) and water (10ml) were added. The product was extracted with CH 2 Cl 2 (2 x 10 mL), dried over MgSO 4, and the solvent removed in vacuo. The residue was pump dried to give Mu-Phe-Hph-γ-CO 2 H (0.30g, 60 wt%).

Example 9

Synthesis of cysteine protease inhibitor containing γ-aminophosphonate as EWG.

The synthesis of diethyl (S) -4- (4-morpholinecarbonyl-phenylalanyl) amino-6-phenylhexanephosphonate (MuPhe-Hph-γ-SO 2 Ph) was performed as follows. To a solution of tetraethyl methylene diphosphonate (2.00 g, 6.94 mmol) in THF (30 mL) was added sodium hydride (0.278 g of a 60 wt% dispersion in mineral oil, 6.94 mmol). The mixture was effervescent, then cleared. After 5 minutes, a solution of Boc-HphH (1.83 g, 6.94 mmol) in THF (5 mL) was added. The mixture was stirred for 1 h, and then 1M HCl (20 mL) was added. The product was extracted into ethyl acetate (50 mL), washed with saturated NaHCO 3 (20 mL), NaCl (10 mL), dried over MgSO 4, filtered and evaporated to dryness to give 2.46 g (89%) of diethyl (S) intermediate. - (E) 4-tert-butoxycarbonylamino-6-phenyl-2-hexenephosphonate. To a solution of this material in CH 2 Cl 2 (3 mL) was added 10 mL of a 4.0 M solution of HCl in dioxane. The mixture was stirred at room temperature for 1.5 h, then the solvents were removed in vacuo and the residue dissolved in methanol (10 mL). The solution was poured into ether (400 mL) and the precipitate collected on a Buchner funnel, washed with ether (2 x 20 mL) and dried on the pump to give 1.25 g (60%) of diethyl (S) - (E) -4 hydrochloride intermediate. -amino-6-phenyl

2-hexenephosphonate. To a solution of Mu-PheOH (1.04 g, 3.74 mmol) in THF (15 mL) at -10 ° C was added 4-methylmorpholine (0.412 mL, 3.74 mmol) and isobutyl chloroformate (0.486 mL, 3.74). mmol). The mixed anhydride was stirred for 5 minutes, and then a solution of (S) - (E) 4-amino-6-phenyl-2-hexenephosphonate hydrochloride (1.25 g, 3.74 mmol) in DMF (5 mL) was added. and

4-methylmorpholine (0.412 mL, 3.74 mmol). The mixture was stirred for 1 h, and then ethyl acetate (50 mL) was added. The solution was washed with 1M HCl (25 mL), saturated NaHCO 3 (25 mL) and NaCl (10 mL), dried over MgSO 4, filtered, and evaporated to dryness. The product was treated with CH ₂ Cl ₂ / ether / hexane (315 mL in 15: 200: 100 ratio) to an oil which solidified on drying in vacuo to give 1.44 g (69%) of diethyl (S) - (E). 4- (4-Morpholinecarbonylphenylalanyl) amino-6-phenyl-2-hexenephosphonate. This material (0.85 g) was dissolved in ethanol (10 mL) and transferred to a Parr bottle of 5 wt. palladium on coal. The solution was reduced in a Parr apparatus for 36 h, and then filtered through diatomaceous earth and the solvent removed in vacuo to give 0.66 g (76%) of the final product as an oil. TLC: (5% MeOH / CH ₂ Cl ₂ ) R _f = 0.27.

Example 10

Synthesis of a cysteine protease inhibitor containing γ-aminoamide as EWG.

Synthesis of (S) -3- (4-morpholinecarbonylphenyl-alanyl) -amino-6-phenylhexanamide (Mu-PheHph-γ-ΑΜΒζΙ). Wadsworth-Emmons reagent (diethylbenzylamidocarbonyl methylene phosphonate) was prepared in two steps; the first was hydrolysis of triethylphosphonoacetate to diethylphosphonoacetic acid, which was then dissolved in ethyl acetate at 0.2 M concentration, then equivalent of benzylamine, 0.1 equivalent of 4-dimethylaminopyridine and 1 equivalent of dicyclohexylcarbodiimide were added.

To a solution of this Wadsworth-Emmons reagent (2.59 g, 9.08 mmol) in THF (40 mL) at 0 ° C was added sodium hydride (0.363 g 60 wt% dispersion in mineral oil, 9.08 mmol). The mixture was stirred at room temperature for 15 minutes, and then Bochomophenylalaninal (2.39 g, 9.08 mmol) in THF (10 mL) was added. The mixture was stirred for 1 h, and then 1M HCl (30 mL) was added. The product was extracted into ethyl acetate (100 mL), washed with saturated NaHCO 3 (50 mL), NaCl (30 mL), dried over MgSO 4, filtered, concentrated and crystallized from ether / hexane to give 1.81 g (51% wt.). Benzyl (S) - (E) 3-tert-butoxycarbonylamino-6-phenyl-2-hexenamide. This material was dissolved in CH ₂ Cl ₂ (5 mL), and then HCl / dioxane (10 mL of a 4.0 M solution) was added. The mixture was then 3 h ·· «···

...... ......

stirred at room temperature. The solvents were removed in vacuo and the residue dissolved in methanol (5 mL) and poured into ether (300 mL). The benzyl (S) - (E) 3-amino-6-phenyl-2-hexenamide hydrochloride intermediate was then separated as an oil by 82 wt. yield (1.25 g). To a solution of Mu-PheOH (1.05 g, 3.78 mmol) in THF (15 mL) at -10 ° C was added 4-methylmorpholine (0.416 mL, 3.78 mmol) and isobutyl chloroformate (0.490 mL, 3 mL). 78 mmol). The mixture was stirred for 10 minutes, and then a solution of benzyl (S) - (E) 3-amino-6-phenyl-2-hexenamide hydrochloride (1.25 g, 3.78 mmol) in THF (3 mL) was added and

4-methylmorpholine (0.416 mL, 3.78 mmol). The mixture was stirred for 45 minutes, and then ethyl acetate (40 mL) was added. The solution was washed with 1M HCl (10 mL), saturated NaHCO 3 (10 mL), NaCl (5 mL), dried over MgSO 4, filtered and evaporated to dryness. Benzyl (S) (E) -3- (4-morpholinecarbonylphenylalanyl) amino-6-phenyl-2-hexenamide intermediate was precipitated from CH 2 Cl 2 / ether in 56 wt. yield. This material (0.48 g, 0.865 mmol) was dissolved in ethanol (10 mL) and transferred to a 5% wt. palladium on coal. The mixture was reduced in a Pan apparatus for 4 h, then filtered through diatomaceous earth and the solvent removed in vacuo. The final product (Mu-Phe-Hph-γ-AMBzl) was crystallized from ethanol / hexane to give 0.25 g (52 wt%). TLC: (50% ethyl acetate / hexane) R f = 0.45.

Example 11

Synthesis of a cysteine protease inhibitor containing γ-aminoamide as EWG.

Synthesis of phenyl (S) -3- (4-morpholinecarbonylphenylalanyl) -amino-6-phenylhexanamide (Mu-Phe-Hph-γ-AMPh). To a solution of Mu-Phe-Hph-γ-CHCl 3 (0.30 g, prepared according to Example 8) in THF (5 mL) at -10 ° C was added triethylamine (90 µΐ, 1 eq.) Followed by isobutyl chloroformate (0.083). mL, 1 eq). After 5 min aniline (0.058 mL) was added, the cooling bath was removed and the mixture stirred at room temperature for 2 h. CH 2 Cl 2 (30 mL) was then added and the solution was washed with 1M HCl and saturated NaHCO 3 (10 mL each), dried with MgSO 4. 4, filtered and the solvent was removed in vacuo. The residue was layered with ether, filtered and dried in vacuo to give 0.29 g (85%) of Mu-Phe-Hph-γ-AMPh. TLC (10% MeOH / CH ₂ Cl ₂ ) R _f = 0.70, strong UV absorption (254 nm), I ₂ .

r -— r · · · · ················

Example 12

Synthesis of an eystein protease inhibitor containing γ-aromatics as EWG.

Synthesis of (S) -4-aminophenyl-3- (4-morpholino-carbonylphenylalanyl) amino-5-phenylpentane hydrochloride (Mu-Phe-hPhe-γ-C 6 H 11 N 4 HCl).

Triphenylphosphine (38.17 g, 0.146 mol) and 4-nitrobenzyl chloride (25g, 0.146 mol) were dissolved in CH 3 CN (100 mL) and heated to boiling for 2 h. After cooling to room temperature, the reaction mixture was diluted with ether (300 mL), the white precipitate filtered off, washed with ether (200 mL) and dried in vacuo to give 53.3 g (84% wt).

4-nitrobenzyltriphenylphosphonium chloride as one spot on TLC: (Rf = 0.71, 4: 1: 1 butanol: acetic acid: water).

1 H-NMR (d 6 -DMSO): 5.40-5.50 (2H, d, CH ₂ P, J 20Hz); 7.20-7.40 (2H, dd, aromatics); 7.40-7.80 (12H, m, aromatics), 7.90-8.00 (3H, m, aromatics), 8.10-8.20 (2H, d, aromatics).

To a stirred suspension of 4-nitrobenzyltriphenylphosphonium chloride (10.02 g, 23.1 mmol) in CH 2 Cl 2 (100 mL) was added 4-methylmorpholine (2.54 mL, 23.1 mmol). After dissolution of the solid, Boc-HphH (4.04 g, 15.4 mmol) was added. After 24 h, the reaction mixture was diluted with CH 2 Cl 2 (200 mL) and filtered. The filtrate was washed with 1M HCl (200 mL), saturated NaHCO 3? (200 mL), dried over MgSO 4, filtered, and concentrated in vacuo to give 4.00 g of the crude intermediate, a portion of which was purified by NMR chromatography (10-30% ethyl acetate / hexane gradient). (S) -1-butoxycarbonyl-3-amino-1- (4-nitrophenyl) -5-phenyl-1-pentene. TLC: (30% EtOAc / hexane) R f = 0.49. To a solution of this material (2.76 g,

A solution of anhydrous p-toluenesulfonic acid (2.76 g, 16.0 mmol) in ether (10 mL) was added in ether (25 mL). The reaction mixture was stirred for 16 h, filtered, the solid washed with ether (25 mL) and dried in vacuo to give 2 g (61%) of (S) 3-amino-1- (4-nitrophenyl) -5-phenyl- l-ene with a single spot on TLC: (Rf = 0,49,10 vol.% MeOH / CH ₂ Cl _2).

To a solution of Mu-PheOH (1.29 g, 4.63 mmol) in THF (20 mL) was added 4-methylmorpholine (0.51 mL, 4.63 mmol) and isobutyl chloroformate (0.61 mL, 4.63 mmol). ). After 3 minutes a solution of (S) 3-amino-1- (4-nitrophenyl) -5-phenyl-1-pentene hydrochloride prepared by deprotection with HCl / dioxane precursor (S) 3-tert-butoxycarbonylamino-1- (4-nitrophenyl) was added. 5-Phenyl-1-pentene (1.34 g, 4.20 mmol) in CH 2 Cl 2 (20 mL) and then

4-methylmorpholine (0.51 mL, 4.63 mmol). The mixture was stirred and warmed to room temperature overnight, and then diluted with CH 2 Cl 2 (100 mL), washed with 1M HCl (200 mL), saturated NaHCO 3 (200 mL), dried over MgSO 4, filtered and concentrated in vacuo to give a yellow oil. This material was crystallized from ClLCL / ether (2: 100.20 mL) to give 1.00 g (40% wt) of (S) -3- (4-morpholinecarbonylphenylalanyl) amino-1- (4-nitrophenyl) -5 -phenyl-1-pentene as a 4: 1 E / Z mixture. This material (0.27 g, 0.49 mmol) was dissolved in ethanol (50 mL), transferred to a Parr bottle with 5 wt% palladium on carbon (0.10 g). The mixture was reduced in a Parr apparatus for 8 h, then filtered and the solvent removed in vacuo. The residue was dissolved in 4: 1 ether / CILCL (100 mL) and HCl / dioxane (0.136 mL of 4.0 M solution) was added. The product, Mu-Phe-Hphy-C0 NIT HCl, was filtered off and dried in vacuo. Yield = 0.15 g (54 wt%). TLC: (10% methanol / CH ₂ Cl ₂ ) R f = 0.31.

Example 13

Synthesis of an eysteine protease inhibitor containing β-aminosulfone as EWG.

Synthesis of (S) -2- (4-morpholinecarbonylphenyl-alanyl) amino-4-phenyl-1-phenylsulfonylbutane (Mu-Phe-HPh-P-SChPh). The preparation of Boc-homophenylalaninol (Boc-Hph-β-ΟΗ) and (S) -2-tert-butoxycarbonylamino-1-methanesulfonyloxy-1-phenylbutane (Boc-Hph-β-Os or Boc-homophenylalaninol mesylate) was performed according to Spaitenstein, Carpino, Miyake and Hopkins (see above). To a solution of Boc-homophenylalanine (10.29 g, 36.84 mmol) in THF (100 mL) at -10 ° C was added 4-methylmorpholine (4.05 mL,

36.84 mmol) and isobutyl chloroformate (4.78 mL, 36.84 mmol). The solution was stirred for 10 minutes and then filtered. The filtrate was carefully added to a stirred solution of sodium borohydride (2.77 g, 73.67 mmol) in water (100 mL) at 0 ° C. The mixture was stirred for 30 minutes, and then saturated NaHCl 3 (200 mL) was added. The product was extracted with CH 2 Cl 2 (2x100 mL), dried over MgSO 4, filtered, and the solvent removed in vacuo to give 9.78 g (100%) of Boc-homophenyl-alaninol. TLC: (30% ethyl acetate / hexane) R f = 0.15. This material (5.83 g, 21.97 mmol) was dissolved in CH 2 Cl 2 (150 mL), cooled to 0 ° C, and methanesulfonyl chloride (4.15 mL, 53.71 mmol) and triethylamine (9.24 mL, 66.3 mmol). The mixture was stirred for 30 minutes and then stirred

water (100 mL) was added with vigorous stirring. The organic layer was separated, dried over MgSO ₄ , filtered and the solvent removed in vacuo to give 7.31 g (97%) of the material. TLC: (30% ethyl acetate / hexane) R f = 0.21. The corresponding Boc-Hph-β-benz benzenesulfonate (ester) was prepared by the same procedure.

To a solution of thiophenol (0.653 mL, 6.36 mmol) in THF (5 mL) was added sodium hydride (0.254 g, 6.36 mmol as a 60 wt% dispersion in mineral oil. The mixture was stirred for 10 minutes and then added Boc-homo-phenylalaninol-benzenesulfonate (2.58 g, 6.36 mmol) in THF (5 mL) The solution was stirred at room temperature for 10 min, then methanol (2 mL) was added and the mixture was heated at reflux for 1 h. The solution was cooled, diluted with 1M NaOH (25 mL), extracted with CH 2 Cl 2 (100 mL), dried over MgSO ₄ , filtered, and the solvent was removed in vacuo, the residue was dissolved in CH 2 Cl 2 (35 mL) and cooled to 0 ° C.

To the solution was added 4-chloroperbenzoic acid (3.71 g, 13.99 mmol, peracid content 65 wt%). The mixture was stirred for 1 h and then 10 wt. NaOH (35 mL) and saturated NaHSO 3 (35 mL). The mixture was extracted with CH 2 Cl 2 (3x50 mL), dried over MgSO ₄ , filtered, and the solvent removed in vacuo to give waxy (S) -2-tert-butoxycarbonylamino-4-phenyl-1-phenylsulfonylbutane. TLC: (30% ethyl acetate / hexane) R f = 0.32. This material (1.25 g) was dissolved in CH 2 Cl 2 (5 mL) and HCl / dioxane (5 mL of 4.0M solution) was added. The mixture was stirred at room temperature for 2 h and then poured into ether (200 mL) to give an oily residue. The solvent over oil was discarded and the residue redissolved in CH 2 Cl 2 (10 mL) and poured into ether (200 mL). The intermediate, (S) -2-amino-4-phenyl-1-phenylsulfonylbutane hydrochloride precipitated. The solid was filtered off and dried in vacuo to give 0.40 g (38% yield from Boc-homophenylalaninol benzenesulfonate).

To a solution of Mu-PheOH (0.342 g, 1.23 mmol) in THF (10 mL) was added at -10 ° C

4-methylmorpholine (0.135 mL, 1.23 mmol) and isobutyl chloroformate (0.159 mL, 1.23 mmol). The mixture was stirred for 10 minutes, and then (S) -2-amino-4-phenyl-1-phenylsulfonylbutane hydrochloride (0.40 g, 1.23 mmol) and 4-methylmorpholine (0.135 mL, 1.23 mmol) were added. . The mixture was stirred for 45 min and then 1M HCl (15ml) was added. The product was extracted with ethyl acetate (30 mL), washed with saturated NaHCO ₃ , (15 mL), saturated NaCl (15 mL), dried over MgSO ₄ , filtered, and the solvent was removed in vacuo. The final product Mu-Phe-Hph 4 -SChPh was obtained in 100 wt. yield (0.68 g).

•!> · · · · · ·

Example 14

Synthesis of a cysteine protease inhibitor containing β-aminosulfone as EWG.

Synthesis of (S) -2-tert-butoxycarbonylamino-4-phenyl-1- (1'-trimethylsilylethyl) -sulfonylbutane (Boc-Hph 4 -SO ₂ CH ₂ CH ₂ TMS). To a solution of 2-trimethylsilylethanethiol (0.86 g, 6.41 mmol), the synthesis was described by Aerson, Ranasinghe, Palmer and Fuchs. above, in THF (10 mL) was added sodium hydride (0.256 g, 6.41 mmol as a 60 wt% dispersion in mineral oil). The mixture was stirred for 10 minutes, and then Boc-homophenylalaninol mesylate (2.00 g, 5.82 mmol, synthesis described in Example 13) was added. The solution was stirred for 2 h, and then ethyl acetate (50 mL) was added. The solution was washed with 30 mL 1M HCl, saturated NaHCO 3 and saturated NaCl (30 mL each), dried over MgSO 4, filtered, and the solvent was removed in vacuo to afford (S) -2-tert-butoxycarbonylamino-4-phenylbutyltrimethylsilylethylsulfide intermediate. TLC: (5% ethyl acetate / hexane) R f = 0.22. This material was dissolved in CH 2 Cl 2 (50 mL), cooled to -10 ° C, and 4-chloroperbenzoic acid (3.24 g, 12.22 mmol, 65 wt% peracid) was added. The mixture was stirred overnight, the suspension was filtered, and aqueous NaHSCl 3 (40 mL) and saturated NaICO 3 (50 mL) were carefully added to the filtrate. The organic phase was separated, dried over MgSO 4, filtered, and the solvent removed in vacuo to give the product Boc-Hph-βSO 2 CH 2 CH 2 CH 2 TMS in quantitative yield from mesylate. TLC: (30% ethyl acetate / hexane) R f = 0.49.

Example 15

Synthesis of a cysteine protease inhibitor containing β-aminosulfone as EWG.

Synthesis of (S) -2- (4-morpholinecarbonylphenylalanyl) -amino-1-chloromethylsulfonyl-4-phenylbutane (Mu-Phe-Hph-p-SO ₂ CH ₂ Cl). To a solution of Boc-Hph 4 -SO ₂ CH ₂ CH ₂ TMS (0.90 g, 2.18 mmol, see Example 14) in THF (2 mL) was added tetrabutylammonium fluoride (8.7 mL of 1.0 M THF solution) and several molecular sieves. The mixture was stirred at room temperature overnight, and then bromochloromethane (5 mL) was added. The mixture was heated to boiling for 1 h, cooled and the volatiles removed in vacuo. The residue was dissolved in ethyl acetate (75 mL), washed with 1M HCl (50 mL), dried over MgSO 4, filtered, and the solvent was removed in vacuo. The residue - crude (S) 2-tert-butoxycarbonyl amino-1-chloromethylsulfonyl-4-phenylbutane - was dissolved in ether (3 mL) and an anhydrous 4-toluenesulfonic acid solution was added. acid (0.80 g, 4.70 mmol) in ether (3 mL). The mixture was stirred at room temperature overnight, and then ether (100 mL) was added. The solid intermediate, (S) -2-amino-1-chloromethylsulfonyl-4-phenylbutane 4-toluenesulfonate (TsOH Hph-p-SCLCH 3 Cl), was filtered off, washed with ether (2 x 20 mL) and dried in vacuo to give 0.193 g (24% w / w of Boc-Hph-P-SChCITClFTMS).

To a solution of Mu-PheOH (0.109 g, 0.392 mmol) in THF (3 mL) at -10 ° C was added 4-methyhnorpholine (43 µΐ, 0.392 mmol) and isobutylchlorophonate (51 µΐ, 0.392 mmol). The mixture was stirred for 10 minutes, and then TsOH.Hf-p-SChCl2 Cl (0.17 g, 0.392 mmol) and 4-methylmorpholine (43 μΐ, 0.392 mmol) were added. The mixture was stirred for 45 minutes, and then ethyl acetate (20 mL) was added. The solution was washed with 1M HCl, saturated NaHCO 3 and NaCl (2 mL each), dried over MgSO 4, filtered, and the solvent removed in vacuo to give the final product, Mu-Phe-Hph-P-SChCl FCl (90 mg, 48% wt). .)

Example 16

Synthesis of a cysteine protease inhibitor containing α-aminosulfone as EWG.

Synthesis of 1- (tert-butoxycarbonyl) amino-2-methyl-1-phenylsulfonylpropane (Boc-Vala-SChPh). To a stirred suspension of t-butyl carbamate (2.34 g, 20 mmol) and sodium benzenesulfinate (3.28 g, 20 mmol) in water (20 mL) was added a solution of isobutyraldehyde (2.00 mL, 22 mmol) in formic acid ( 5 ml). The mixture was stirred at room temperature overnight, then the precipitate formed was filtered off and washed with water (2 x 50 mL) and crystallized from isopropanol / water to give 4.72 g (75%) of the product.

Example 17

Synthesis of a cysteine protease inhibitor containing α-aminosulfone as EWG.

Synthesis of 1-benzyloxycarbonylamino-3-phenyl-1-phenylsulfonylpropane (Z-Hph-α-SChPh).

To a suspension of sodium benzenesulfinate (10 g, 60.9 mmol) and benzyl carbamate (9.21 g,

60.9 mmol) in water (40 mL) was added hydrocinnamaldehyde (8.8 mL, 67 mmol) in formic acid (10 mL). The mixture was heated at 70 ° C for 1 h and then allowed to warm to room temperature overnight. The product crystallized, was filtered off and recrystallized from hot isopropanol to give 23 g (100%) of material. TLC:

(30% v / v ethyl acetate / hexane) R f 0.37.

Example 18

Synthesis of a cysteine protease inhibitor containing α-aminosulfone as EWG.

Synthesis of (R) -1- (4-morpholinecarbonylphenylalanyl) amino-3-phenyl-1-phenylsulfonylpropane and (S) -1- (4-morpholinecarbonylphenylalanyl) amino-3-phenyl-1-phenylsulfonylpropane (MuPheHph-α-SChPh, separated epimers).

Method A: To Z-Hph-α-SChPh (1.0 g, 2.44 mmol) was added 30 wt. hydrogen bromide in acetic acid (5 mL). After 30 minutes, the mixture was diluted with ether (300 mL), filtered, washed with ether (2 x 30 mL) and dried in vacuo to give 0.74 g (86%) of 1-amino-3-phenyl-1-phenylsulfonylpropane hydrobromide (HBrHpha). -SO2Ph). To a solution of Mu-PheOH (0.64 g, 2.3 mmol) in THF (15 mL) was added 4-methylmorpholine (0.302 mL, 2.3 mmol) and isobutyl chloroformate (0.312 mL, 2.3 mmol). The mixture was stirred for 10 minutes, and then HBr Hph-α-SO ₂ Ph (0.74 g, 2.1 mmol) and 4-methylmorpholine (0.302 mL, 2.3 mmol) were added. After 45 minutes, the mixture was diluted with ethyl acetate (30 mL), washed with 15 mL of 1M HCl, saturated with Na 2 CO 4, NaCl (15 mL of each), dried over MgSO 4, filtered and the solvent removed in vacuo to give 0.75 g ( 65 wt% of the product, Mu-Phe-Hph-α-SChPh.

Method Β: To a solution of phenylalanine amide hydrochloride (10 g, 50 mmol) in DMF (50 mL) and CH 2 Cl 2 (50 mL) was added triethylamine (13.9 mL, 100 mmol) and 4-morpholine carbonyl chloride (5.9 mL, 50 mmol). The mixture was stirred overnight, and then the solvents were removed in vacuo. The residue was dissolved in ethyl acetate (50 mL) and filtered. Ether was added to the filtrate until the solution became cloudy. After 3 days, 7.2 g (80%) of the intermediate, 4-morpholinecarbonylphenylalanine amide (Mu-Phe-NH2) crystallized from the solution. To a solution of Mu-Phe-NH 2 (2.24 g, 8.1 mmol) in formic acid (5 mL) was added hydrocinnamaldehyde (1.17 mL, 8.9 mmol) with stirring. The mixture was stirred for 5 h, and then sodium benzenesulfinate (1.33 g, 8.1 mmol) was added. The mixture was heated to boiling for 5 min and then allowed to cool to room temperature. The solution was then stirred for 3 days, and then 1 volume equivalent of water was added. The product was extracted with CH 2 Cl 2 (3x100 mL), dried over MgSO 4, filtered, and the solvent removed in vacuo.

.:. .:.

The yield of the product - diastereomeric (R) - and (S) -1- (4-morpholine-carbonylphenylalanyl) amino-3-phenylsulfonyl-1-phenylpropane - was 3.9 g (90% by weight). TLC (50% v / v ethyl acetate / CH 2 Cl 2 R f = 0.27 and 0.34).

The diastereomers were separated by flash chromatography on 230-400 mesh silica gel (20-50 v / v ethyl acetate / Q-Ch 2 gradient).

Example 19

Conditions for cathepsin B: 50 mM phosphate, pH 6.0, 2.5 mM EDTA, 2.5 mM DTT. Substrate: [Z-Arg-Arg-AMC] = 50 mM (Km 190 mM). The assay was started at 25 ° C by addition of cathepsin B (final concentration 10 nM) and after 2 min fluorescence at 450 nm with excitation at 380 nm was monitored. A decrease in the rate of substrate hydrolysis was observed following the addition of inhibitor at various concentrations. The test was linear in the study area. The measurement was doubled.

Conditions for cathepsin L: 50 mM acetate, pH 5.5, 2.5 mM EDTA, 2.5 mM DTT. Substrate: (Z-Phe-Arg-AMC) - 5 mM (Km = 2 mM) The assay was started at 25 ° C by addition of cathepsin L (final concentration 1 nM) and after 2 min fluorescence at 450 nm was observed with excitation at 380 The decrease of the rate of hydrolysis of the substrate was observed after the addition of inhibitor with different concentrations and the test was linear in the monitored area.

Conditions for cathepsin S: 50 mM phosphate, pH 6.5, 2.5 mM EDTA, 2.5 mM DTT. Substrate: [Z-Val-Val-Arg-AMC] = 10 mM (Km = 18 mM). The assay was started at 25 ° C by addition of cathepsin S (final concentration 30 µM) and after 2 min fluorescence at 450 nm with excitation at 380 nm was monitored. A decrease in the rate of substrate hydrolysis was observed following the addition of inhibitor at various concentrations. The test was linear in the study area. The measurement was doubled.

The conditions for cruzain were the same as for cathepsin L with the K m of the substrate being 1 mM.

A · A

AAA

AAA A

A A

A A A

Appropriate K values were determined according to the Dixon curve described by Irwin Segel in Enzyme Kintics: Behavior and Analysis of Rapid Equilibrium and Steady-State Enzyme Systems, 1975, Wiley-Interscience Publication, John Wiley & Sons, New York.

The results are shown in Table 2.

Table 2

Inhibitor ..........., cathepsin B (Κ, μΜ) cathepsin L cathepsin S cruzain Mu-Phe-Dl) HphaSO, Ph 3,000 13 5 15 Dec Mu-Phe-hpHAse ₂ Ph « 2.6 Mu-Phe ~ (DL) hpHAse ₂ Ph. • 3.7 Mu-Leu-HphaSO2Ph 16 1.3 1.6 027 Mu-Leu-HphaSO ₂ 54 0.60 4.2 0.76 Mu-Phe-HphaSO ₇ CH _? F 170 1.5 2.8 2.0 Mu-Phe-HphaSO _ř Bzl 77 1.2 5.2 0.92 Mu-Phe-HphaSO3CRj 50 0.18 2.2 0.23 Mu-Phe-HphpSO ₂ Ph 1.100 29 0.94 5.7 Mu-Phe-HphySO ₂ Ph 48 10 0.16 4.3 Phac-Phe-HphvSOjPh »10 0.41 12.5 1.8 Mu-Np2 * HphySO _? 2Np 20 May 0.26 0.53 0 10 Mu-Phe-HphYSO3EtPh 190 0.17 0.082 5.9 Suc-Phe-HphySO ₂ Ph > 1000 1.0 0.07 1.5 MeOSuc-Phe-HphySO ₂ Ph 81 3.2 1.2 4.7 Suc-Np2-HphySO ₂ Ph > 1000 2.3 0.50 0.78 Suc-Np2-HphySO2Np > 1000 0.11 0.24 0.23 ZP-Ala-Phe-HphySO ₂ Ph 52 0.79 3.0 0.54 I O-Ala-Phe-HphySO _? Ph »50 14 11 14 Mu-Tyr-HphySO2Ph - 2.3 9.5 20 May Mu-Phe-HphyCO, Et 1.6 0.48 0.19 0.91 Mu-Phe-HphyCONHPh 2.6 2.0 1.3 0.13 Mu-Phe-HphyCONHBzl »50 19 Dec 30 7.7 Mu-Phe-HphyPO (O ₂ Et) ₂ 17 3.0 1.4 15 Dec Mu-Phe-Hph-γPh-OMe 4.8 0.94 0.37 0.89 Mu-Phe-Hph-yPh-NH »50 2.9 9.1 3.0

• · ·

Example 20 Protease in accordance with the present invention.

Oral agent

A representative solution for oral administration comprises:

cysteine protease inhibitor citric acid monohydrate sodium hydroxide flavor water

Intravenous agent

A representative solution for intravenous administration comprises: cysteine protease inhibitor dextrose monohydrate concentration citric acid monohydrate sodium hydroxide isotonic solution for injection

Tablet formulation

A representative tablet contains:

cysteine protease inhibitor microcrystalline cellulose stearic acid colloidal diatomaceous earth

100 to 1000 mg 105 mg 18 mg residue to 100 ml to 100 mg to isotonic

1.05 mg 0.18 mg residue to 10 ml wt. 73 wt. 25 wt.

% wt.

Industrial applicability

Cysteine protease inhibitors are widely applicable. For example, they can be used to determine the amount of cysteine protease present in a sample and are therefore used in diagnostic kits for cysteine proteases in blood, sap, saliva or other specimens as well as in bacteria, fungi, plants, yeasts, viruses or cultures of mammalian cells. . Therefore, in a preferred embodiment, the sample is tested using a standard protease substrate. After addition of a known concentration of cysteine protease inhibitor, this is allowed to bind to the cysteine protease present. The protease assay is then repeated and the loss of its activity is correlated with a technique known in the art.

Said inhibitors are also suitable for removing or inhibiting contaminating cysteine proteases in a sample. For example, an inhibitor according to the present invention is added to a sample where proteolytic degradation contaminating a cysteine protease is undesirable.

The inhibitor of the present invention can further be coupled to a chromatographic support using techniques known in the art to prepare an affinity chromatography column. If a sample containing an undesired cysteine protease passes through this column, this will be removed.

In a preferred embodiment, the inhibitor is used to inhibit proteases acting in a variety of diseases. Specifically, the inhibition of cathepsins B, L, and S, cruzain, calpaines I and II, and interleukin 1β conversion enzyme. These enzymes are examples of lysosomal cysteine proteases involved in a variety of diseases characterized by tissue loss such as (but not limited to) arthritis, muscular dystrophy, inflammation, tumor invasion, glomerulonephthis, parasitic infection, Alzheimer's disease, dental disease and cancer metastasis. E.g. mammalian lysosomal thiol proteases play an important role in the intracellular degradation of proteins and in the processing of certain peptide hormones. Cathepsin B and L-like enzymes are released by tumors and may be involved in their metastasis, cathepsin L is present in patients in synovial fluid and in transformed tissues. Similarly, the release of cathepsins B and other lysosomal proteases from polymorphonuclear granulocytes is observed in injury and inflammation.

• · · β7 · · · · · ·

ΟΖ ······ ··· ···

Inhibitors in accordance with the present invention find application in a variety of other diseases including but not limited to gingivitis, malaria, kala-azar, filariasis and other infections from bacteria and parasites. These compounds are also available in viral diseases, based on the inhibition of proteases necessary for viral replication. E.g. a number of picomoviruses, including polioviruses, foot and mouth disease viruses and rhinitis viruses, encode cysteine proteases that are necessary for the cleavage of viral polyproteins.

In addition, they are offered for diseases related to interleukin-ΐβ conversion enzyme * (ICE) and the cysteine protease responsible for processing interleukin-ΐβ; eg in the treatment of inflammations and immunological diseases of the lungs, airways, central nervous system and surrounding membranes, eyesight, hearing, joints, bones, connective tissues, cardiovascular system including pericardium, gastrointestinal and urogenital system, skin and mucous membranes, including infectious diseases, where there is an acute infection anywhere in / on the body such as meningitis and alpingitis; complications of infections such as septic shock, multiple sclerosis and adult respiratory distress syndrome; acute or chronic inflammation due to antigens, antibodies and / or complementary deposition: inflammations such as arthritis, chalazion, colon inflammation, encephalitis, endocarditis, glomerulonephritis, hepatitis, myocarditis, pancreatitis, pericarditis, overwhelming injury and vascular inflammation. Immunological diseases include, but are not limited to, T-celis and / macrophages such as acute and chronic hypersensitivity, transplant tissue rejection, host-transplant tissue disease; auto-immune diseases such as diabetes mellitus type I and multiple sclerosis. Thinning of bones and cartilage and diseases of excessive deposition of the extracellular matrix such as interstitial pulmonary fibrosis, cirrhosis, systemic sclerosis and keloid formation are also diseases that can be treated with inhibitors in accordance with the present invention. The inhibitors of the present invention can also be used to treat some tumors that produce IL 1 as an autocrine growth factor and to prevent cachexia associated with some tumors. Cell death is also associated with ICE and the like activity, which can also be treated with inhibitors in accordance with the present invention.

Inhibitors according to the present invention find use as drug promoters. E.g. antibiotics or other drugs may be inactivated by proteolysis of endogenous cysteine protease, resulting in less efficacy or ineffectiveness of the administered drugs. E.g. bleomycin has been shown (an anticancer medicine) to be hydrolyzed by bleomycin hydrolase, a cysteine protease (see Sebti et al., Cancer Res. January)

1991,227-232). Accordingly, the inhibitors of the present invention may be administered to patients in conjunction with other drugs to enhance or increase the activity of the drug, which may be accomplished by simultaneous administration, i.e., by administering a mixture of a cysteine protease inhibitor and a drug, or by administering each agent separately.

Furthermore, cysteine protease inhibitors have been shown to inhibit the growth of bacteria, particularly human pathogenic bacteria (see Bjorck et al., Nature 337: 385 (1989)). Therefore, cysteine protease inhibitors in accordance with the present invention can be used as an antibacterial agent to inhibit or inhibit the growth of certain bacteria.

Cysteine protease inhibitors in accordance with the present invention may also be used to reduce or destroy bacterial cysteine proteases in a host organism.

E.g. staphylococci produce a very active extracellular cysteine protease that breaks down insoluble elastin, potentially contributing to damage to connective tissues in bacterial infections such as septemia, septic arthritis and otitis. See. Potempa et al.,

J. Biol. Chem. 263 (6): 2664-2667 (1988). Therefore, cysteine protease inhibitors in accordance with the present invention can be used in the treatment of bacterial infections to prevent tissue loss. '

Claims (53)

A protease inhibitor comprising a targeting group linked by two chains of carbon atoms to an electron-attracting group, wherein the dissociation constant for protease inhibition by said inhibitor (K i) is at most 100 μΜ. 2. A protease inhibitor containing a directing moiety linked, either directly or through a linker selected from the group consisting of a middle carbon atom or a di-carbon chain, with a sulfone group, wherein the dissociation constant for protease inhibition by said inhibitor (K i) is at most 100 μΜ. 3. A compound of formula I Wherein n is 0 to 13; AB is a linker of the group -C (O) NR ³ -, -CH ² NR ³ -, -C (O) CH 2 - and -NR ³ C (O) -, wherein R ³ is hydrogen or a group defined below; X is a bond, methylene, or -CH ₂ CH (R ⁴ ) -, wherein R ⁴ is hydrogen, alkyl or arylalkyl; Y is -CH (R ⁵ ) - or -NR ⁵ -, wherein R ⁵ is hydrogen or as defined below; Z is - (CH ₂ ) ₂ -, -C (R ⁶ ) (R ⁷ ) - or -N (R ⁷ ) -, wherein R ⁶ is hydrogen or methyl and R ⁷ is defined below; Z ¹ is - (CH ₂ ) ₂ -, -C (R ⁶ ) (R ⁸ ) - or -N (R ⁸ ) -, wherein R ⁶ is hydrogen or methyl and R ⁸ is defined below; R @ ¹ is hydrogen, alkyloxycarbonylalkanoyl, alkyloxycarbonyl, alkanoyl unsubstituted or substituted by a radical selected from the group consisting of carboxyl, alkyloxycarbonyl and heterocycloalkylalkanoylamino, cycloalkylcarbonyl, -. · · · · · 4 4 4 4 4 4 4 4 4 4 4 · 4444 44 444 444 · heterocycloalkylcarbonyl unsubstituted or substituted by a radical selected from the group consisting of hydroxyl, alkyl, alkanoyl, alkyloxycarbonyl, arylalkyloxycarbonyl and heterocycloalkylcarbonyl, arylalkyloxycarbonyl, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, dialkylcarbamoyl, dialkylcarbamoyl, dialkylcarbamoyl, dialkylcarbamoyl, dialkylcarbamoyl, dialkylcarbamoyl or heteroarylsulfonyl; R ⁷ and R ⁸ are independently hydrogen, alkyl unsubstituted or substituted by hydroxyl, amino, alkylamino, dialkylamino, uriedo, mercapto, alkylthio, carboxy, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, alkylsulfonyl and guanidine or protected derivatives thereof, cycloalkyl, cycloalkylalkyl, heteroaiyl, heteroarylalkyl, aryl and arylalkyl unsubstituted or substituted on the aryl ring with one to three radicals selected from the group consisting of hydroxyl, amino, guanidino, halogen, optionally halogen-substituted alkyl, alkyloxy and aryl or protected derivatives thereof, or together with adjacent R ³ or R ⁵ groups form a divalent radical from the group consisting of (C 6-6) methylene and 1,2-phenylenedimethylene unsubstituted or substituted by hydroxy or protected by hydroxy or oxo; and R ² is hydrogen, alkyl unsubstituted or substituted by one or more of the group consisting of amino, halogen, hydroxyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl or protected derivatives thereof, cycloalkyl, cycloalkylalkyl or aryl and arylalkyl groups unsubstituted or substituted on one aryl ring two radicals selected from the group consisting of amino, guanidino, halogen, hydroxyl, optionally halogen-substituted alkyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl or protected derivatives thereof; and pharmaceutically acceptable salts, individual isomers and mixtures of isomers thereof. The compound of claim 3, wherein the dissociation constant of protease inhibition by said inhibitor (K i) is at most 100 µΜ. The compound of claim 3, wherein n is 0 to 5; A-B are linkers of formula -C (O) NR ³ -, Y is -N (R ⁵ ) -, Z is - (CH ₂ ) ₂ - or -C (R ⁶ ) (R ⁷ ) -; Z ¹ is -CH (R ⁸ ) -; R ¹ is hydrogen, (C 3 -C 10) alkyloxycarbonylalkanoyl, (C 1-9) alkoxycarbonyl; (C 2-10) alkanoyl unsubstituted or substituted by a radical from the group C 2-10 (C 2-10) alkanoyl consisting of carboxyl, (C 1-9) alkyloxycarbonyl and hetero (C _4-8 ) cycloalkyl (C 2-10) alkanoylamino, (C 4-9) cycloalkylcarbonyl, hetero (C _4-8 ) cycloalkylcarbonyl unsubstituted or substituted with a hydroxyl group from the skypine (C |. ₅₎ alkyl, (C sjalkanoyl, (Ci. ₅₎ alkyloxycarbonyl, _(C6-iO) aryl (Ci.5) alkyl benzyloxy carbonyl and hetero (C4. _g) cycloalkylcarbonyl, (C-iO) of allyl (C ₅₎ alkyloxycarbonyl, carbamoyl, (C sjalkylkarbamoyl, di (C-5) alkylcarbamoyl, (C6-io) arylcarbamoyl, _(C6-iO) aryl (Ci. ₅₎ alkyIkarbamoyl, (C ₆ .io) of allyl (Ci. ₅₎ alkanoyl, (C ₇ -n) aroyl, (Cj.5) alkylsulfonyl, di (C-5) alkylaminosulfonyl, (C ₆ .jo) arylsulfonyl or hetero (C5 g) aiylsulfonyl; and R ⁷ and R ⁸ are independently (C 3-7) cycloalkyl, (C3-7) cycloalkyl (Ci-5) alkyl, pyridyl, thienyl, furyl, imidazolyl, indolyl, pyridyl (C ^) alkyl, thienyl (C _(^) alkyl, fuiyl (C) ^) alkyl, imidazolyl (C] <,) alkyk indolyl (C ^) alkyl, selected from the group consisting of phenyl, naphthyl, phenyl (C ^) alkyl, naphthyl _(C ^) alkyl, (Cj- ₅ ) alkyl unsubstituted or substituted by a group consisting of mercapto, carboxy, amino, methylthio, methylsulfonyl, carbamoyl, dimethylcarbamoyl, guanidino and hydroxy or a protected derivative thereof, phenyl, naphthyl, phenyl (C 1-6) alkyl, naphthyl (C 1-6) alkyl unsubstituted or substituted in the aryl moiety with one to three substituents selected from the group consisting of hydroxyl, amino, chloro, bromo, iodo, fluoro, methyl, trifluoromethyl, methoxy and phenyl, or protected derivatives thereof, or together with adjacent R ³ or R ⁵ groups form one of the bivalent radicals including (C3-4) methylene and 1,2-phenylenedimethylene unsubstituted or subs substituted by hydroxy or its protected derivatives or oxo; and R ² is (C 1-5) alkyl unsubstituted or substituted by one or two residues of the series consisting of amino, chloro, bromo, fluoro, iodo, hydroxy, and methoxy, or a protected derivative thereof, perhalogen (C 1-5) alkyl, (C3) -7) cycloalkyl, (C3-7) cycloalkyl (Ci. ₅₎ alkyl or phenyl, pentafluorophenyl, naphthyl and phenyl (C ^) alkyl optionally substituted at its aryl ring with one or two radicals from the series comprising amino, chlorine, bromine, fluorine, , hydroxyl, methoxy and halogen unsubstituted or substituted methyl or a protected derivative thereof; and R ⁴ is hydrogen, (C 1-5) alkyl or (C 6-10) aryl (C 1-5) alkyl. ······················· ! • · · · · · · · · · · · · · 0 0 0 00.000 ··· The compound of claim 5, wherein n is 0 to 2; Z is - (Cfbh- or -C (R ⁶ ) (R ⁷ ), except when n is 0, Z is m - (C 6 H 6 -); R ¹ is hydrogen, (C 4-6) alkoxycarbonyl, (C2 (4) alkanoyl unsubstituted or substituted by a group consisting of carboxyl, (C 1-5) alkyloxycarbonyl and hetero (C 4-6) cycloalkyl (C 4-6) alkanoylamino, -C (O) NR ²¹ R ²² wherein R ²¹ and R ^{21 22 taken} together form aza (C _2-4 ) methylene, oxa (C 2-6) methylene or (C 3-7) methylene, (C 4-8) cycloalkylcarbonyl, benzyloxycarbonyl, acetyl, benzoyl or dimethylaminosulfonyl; and R ⁸ and R ⁷ are independently (C 5-6 cycloalkyl, (C 5-6 cycloalkylmethyl, 3-pyridyl, 2-thienyl, 2-furyl, 4-imidazolyl, 3-indolyl, 3-pyridylmethyl, 2-thienylmethyl, 2-furylmethyl, 4-imidazolylmethyl, 3-indolylmethyl, methoxy, acetoxyl, (C 1-6 alkyl unsubstituted or substituted with a mercapto, carboxy, amino, methylthio, methylsulfonyl, carbamoyl, dimethylcarbamoyl, guanidino and hydroxy derivative or a protected group thereof; , phenyl, 1-naphthyl, 2-naphthyl, benzyl, 1-naphthylmethyl, 2-naphthylmethyl and 2-phenylethyl on the aromatic ring unsubstituted or substituted by a radical selected from the group consisting of hydroxy, amino, chlorine, bromine and fluorine or a protected derivative thereof, or together with adjacent R ³ or R ⁵ groups group wherein the (C ₃ -i) methylene and 1,2-phenylenedimethylene unsubstituted or substituted by hydroxy or its protected derivatives, or oxo; and R ² is (C _1-5 ) alkyl unsubstituted or substituted by one or two of the group consisting of amino, chloro, bromo, fluoro and hydroxy, or a protected derivative thereof, perfluoro (C 1-5) alkyl, (C 5-6 cycloalkyl, ( C 5-6 cycloalkylmethyl or one of the following phenyl, naphthyl and benzyl groups unsubstituted or substituted by a radical of the group consisting of aminohydroxy, chloro, bromo or fluoro, or protected derivatives thereof, and R ⁴ is hydrogen or methyl. The compound of claim 4, wherein n is 0 to 1; Z is -C (R ⁶ ) (R ⁷ ) -; R ¹ is hydrogen, tert-butoxycarbonyl, benzyloxycarbonyl, acetyl, 3-carboxypropionyl, 3-methoxycarbonylpropionyl, biotinylaminohexanoyl, phenylacetyl, benzoyl, dimethylaminosulfonyl, benzylsulfonyl, 1-piperazinylcarbones 1,4-methylpiperazin-1-ylcarbonyl or 4-morpholinylcarbonyl; R ⁸ is butyl, 2-phenylethyl, 2-methylsulfonylethyl, 2-tert-butoxycarbonylmethyl, 2-tert-butoxycarbonylethyl, 73 ··············· 4-tert-butoxycarbonylaminobutyl, 4-benzoylaminobutyl or benzyloxymethyl; R ² is methyl, trifluoromethyl, unsubstituted or substituted phenyl, 2-naphthyl or 2-phenylethyl; R ⁴ is hydrogen; and R ⁷ is 3-pyridylmethyl, 2-thienylmethyl, 2-furylmethyl, 4-imidazolylmethyl, 3-indolylmethyl, (C _1-5 ) alkyl unsubstituted or substituted by one of mercapto, carboxy, amino, methylthio, methylsulfonyl, carbamoyl, dimethylcarbamoyl, guanidino and hydroxy, or a protected derivative thereof, benzyl, 1-naphthylmethyl, 2 -naphthylmethyl and 2-phenylethyl on the aryl ring unsubstituted or substituted by a radical selected from the group consisting of hydroxy, amino, chlorine, bromine and fluorine or a protected derivative thereof, or together with adjacent groups R ³ or R ⁵ form one of the following bivalent radicals (C N-methylene and 1,2-phenylenedimethylene unsubstituted or substituted by hydroxy, or a protected derivative thereof, or oxo. The compound of claim 7, wherein n is 0; R ³ , R ⁵ and R ⁶ are hydrogen; R ¹ is hydrogen, tert-butoxycarbonyl, benzyloxycarbonyl, biotinylaminohexanoyl, benzyl, o 1-piperazinylcarbonyl, 4-methylpiperazinylcarbonyl or 4-morpholinylcarbonyl; R is butyl, 2-phenylethyl or 2-methylsulfonylethyl; R ² is phenyl, 1-naphthyl or 2-phenylethyl and R ⁷ is (C 1-5) alkyl, 2-methylsulfonylethyl, substituted or unsubstituted benzyl, 1-naphthylmethyl, 2-naphthylmethyl, 3-pyridinylmethyl or 2-methylsulfonylethyl. The compound of claim 8, wherein R ¹ is 1-piperazinylcarbonyl, 4-methyl-1-piperazinecarbonyl or 4-morpholinylcarbonyl; R ⁸ is 2-phenylethyl; R ^{2 is} phenyl naphth-2-yl and R ⁷ is substituted or unsubstituted benzyl, 1-naphthylmethyl or 2-naphthylmethyl. 1 g The compound of claim 9, wherein X is a bond, R is 4-morpholinylcarbonyl, R is 2-phenylethyl, R ² is phenyl, and R ⁷ is benzyl which is N ² - (4-morpholinylcarbonyl) N ¹ - (3 - phenyl-1-phenylsulfonylpropyl) -L-phenylalaninamide. The compound of claim 9, wherein X is methylene, R ¹ is 4-morpholinylcarbonyl, R ⁸ is 2-phenylethyl, R ² is phenyl and R ⁷ is benzyl which is N ² - (4-morpholinylcarbonyl) N ¹ - ( 3-Phenyl-1S-phenylsulfonylmethylpropyl) -L-phenylalaniamide. ·· ···· —y Λ · · · ^w 74 ··············· The compound of claim 9, wherein X is -CH 2 CH (R ⁴ ), wherein R ⁴ is hydrogen, R ¹ is 4-morpholinylcarbonyl, R ⁸ is 2-phenylethyl, R ² is 2-naphthyl and R ⁷ is 2-naphthylmethyl which is N ² - (4-morpholinylcarbonyl) -N ¹ - {3-phenyl-1 S- [2- (2-naphthylsulfonyl) ethyl] propyl} b- (2-naphthyl) -L-alaninamide. A compound according to claim 9, wherein X is -CH 2 CH (R ⁴ ), wherein R ⁴ is hydrogen, R ¹ is 4-morpholinylcarbonyl, R ⁸ is 2-phenylethyl, R ² is phenyl and R ⁷ is 4-hydroxybenzyl, is ^2- (4-morpholinylcarbonyl) -N ^1- {3-phenyl-1 S- [2- (2-naphthylsulfonyl) ethyl] propyl} L-tyrosinamide. A compound according to claim 9, wherein X is -CH 2 CH (R ⁴ ), wherein R ⁴ is hydrogen, R ¹ is 4-morpholinylcarbonyl, R ⁸ is 2-phenylethyl, R ² is phenyl and R ⁷ is benzyl which is N ^2- (4-Morpholinylcarbonyl) -N ^1- {3-phenyl-1 S- [2- (2-naphthylsulfonylethylethyl) propyl} L-phenylalaninamide. 15. A compound of formula II-B n II wherein n is 0 to 13; AB is a linker of the group -C (O) NR ³ -CH ² NR ³ -, -C (O) CH 2 - and -NR ³ C (O) -, wherein R ³ is hydrogen or a group defined below; Y is -CH (R ⁵ ) - or -NR ⁵ -, wherein R ⁵ is hydrogen or as defined below; Z is - (CH 2) 2 -, -C (R ⁶ ) (R ⁷ ) - or -N (R ⁷ ) -, wherein R ⁶ is hydrogen or methyl and R ⁷ is defined as may; Z ¹ is - (CH ₂ ) ₂ -, -C (R ⁶ ) (R ⁸ ) - or -N (R ⁸ ) -, wherein R ⁶ is hydrogen or methyl and R ⁸ is defined as mze; R ¹ is hydrogen, alkyloxycarbonylalkanoyl, alkyloxycarbonyl, alkanoyl unsubstituted or substituted by a radical selected from the group consisting of carboxyl, alkyloxycarbonyl and heterocycloalkylalkanoylamino, cycloalkylcarbonyl, heterocycloalkycarbonyl unsubstituted or alkyloxyalkyloxycarbonyl and alkyloxycarbonyl, alkyl, alkoxy, and alkoxy, aylalkyloxycarbonyl, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, arylcarbamoyl, arylalkylcarbamoyl, arylalkanoyl, aryloyl, alkylsulfonyl, dialkylaminosulfonyl, arylsulfonyl or heteroarylsulfonyl; R ⁷ and R ⁸ are independently hydrogen, alkyl unsubstituted or substituted with hydroxyl, amino, alkylamino, dialkylamino, uriedo, mercapto, alkylthio, carboxy, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, alkylsulfonyl and guanidino or a protected derivative thereof, cycloalkyl, cycloalkyl , heteroaryl, heteroarylalkyl, aryl and arylalkyl unsubstituted or substituted on the aryl ring by one to three radicals selected from the group consisting of hydroxyl, amino, guanidino, halogen, optionally halogen-substituted alkyl, alkyloxy and aryl groups or protected derivatives thereof, or together with adjacent R ³ or R ⁵ groups form a divalent radical which is ((H-methylene and 1,2-phenylenedimethylene unsubstituted or substituted by hydroxy or protected by hydroxy or oxo; R ⁹ is cyano, -C (O) OR ^I0, -P (O) (OR ^t0) 2, -S (O) ^{(NR, 0)} R ^I0 C (O) ^R, -S (O) R ⁿ , -C (O) NR ^, 2R * ³ -S (O) 2 NR ¹² R ¹³ , -C (O) NHR ¹⁴ or -S (O) ₂ NHR ¹⁴ wherein R ¹⁰ is independently hydrogen, alkyl unsubstituted or substituted with one or more amino, halogen, hydroxyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl or protected derivatives thereof, cycloalkyl, cycloalkylalkyl or aryl and arylalkyl groups which are unsubstituted or substituted by one or two radicals on the aryl ring containing amino, halogen, hydroxyl, halogen-unsubstituted or substituted alkyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl, or protected derivatives thereof, R ¹¹ is hydrogen, alkyl, perfluoroalkyl, cycloalkyl, cycloalkylalkyl, perfluoraryl, perfluorarylakyl or aryl and arylalkyl groups which are on an aryl ring optionally substituted by one or two radicals from the group A; amino-containing, halogen, hydroxy, halogen-unsubstituted or substituted alkyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl or protected derivatives thereof; R ¹² and R ¹³ are independently hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl or arylalkyl, and R ¹⁴ is -C (O) OR ¹⁰ , wherein R ¹⁰ is as defined above or one of the substituents of formulas (a) and (b) wherein n, A, Β, Y, Z, R ¹ and R ¹⁰ are as defined above, and pharmaceutically acceptable salts, individual isomers, and mixtures of isomers thereof. The compound of claim 15, wherein n is 0 to 5; AB is a linker of the formula -C (O) NR ³ -, Y is -N (R ⁵ ) -; Z is - (CH 2) 2 - or -C (R ⁶ ) (R ⁷ ) -; Z ¹ is -CH (R ⁸ ) -; R ¹ is independently hydrogen, alkyloxycarbonylalkanoyl of overall 3 to 10 carbon atoms (C-9) alkoxycarbonyl, (C2.i ₀₎ alkanoyl unsubstituted or substituted by one radical selected from the group consisting of carboxy, (C | .9) alkyloxycarbonyl and hetero ( C 4-8) cycloalkyl (C ₂ _ i ojalkanoylaminoskupina, (C4_9) cycloalkylcarbonyl, hetero (C4-8) cycloalkylcarbonyl unsubstituted or substituted with one radical selected from the group consisting of hydroxyl, (C sjalkyl, (C, s) alkanoyl, (C -5) alkyloxycarbonyl, (C 6-10) aryl (C 1-5) alkyloxycarbonyl and hetero (C 4-8) cycloalkylcarbonyl, (C 6-10) alkyl (C 1-5) alkyloxycarbonyl, carbamoyl, (C 1-5) alkylcarbamoyl, di (C 1-8) 5) alkylcarbamoyl, _(C6-IO) arylcarbamoyl, _(C6 .io) aryl (C.) ₅₎ alkylcarbamoyl, (C ₆ .io) aryl (C]. ₅₎ aIkanoyl, (C ₇ -II) aroyl, (C 1-6 alkylsulfonyl, di (C 1-5) alkylaminosulfonyl, (C 6-10) aryl sulfonyl or hetero (C 5-8) arylsulfonyl; and R ⁷ and R ⁸ are independently (C3 _'7) cycloalkyl, (C3-7) cycloalkyl (Ci-5) alkyl, pyridyl, thienyl, fuiyl, imidazolyl, indolyl, pyridyl (C | ^) alkyl, thienyl (C ^) alkyl, furyl (C 1-6) alkyl, imidazolyl (C _1-6 ) alkyl, indolyl (C 1-6) alkyl, (C 1-6) alkyl, (C 2-6) alkyloxy and (C 1-5) alkanoyloxy unsubstituted or substituted with a radical from the series consisting of mercapto, carboxy, amino, methylthio, methyisulfonyl, carbamoyl, dimethylcarbamoyl, guanidino and hydroxy or a protected derivative thereof, phenyl, naphthyl, phenyl (C 1-6) alkyl, naphthyl (C 1-6) alkyl, which are not or are not substituted in the aryl moiety by one to the following three radicals from the group consisting of hydroxy and amino, chloro, bromo, iodo, fluoro, methyl, trifluoromethyl, methoxy and phenyl or their protected derivatives, or together with adjacent R ³ or R ⁵ groups one of the bivalent radicals from the group, which consists of / Ca 2 -methylene and 1,2-phenylenedimethylene not substituted or unsubstituted by hydroxy or its protected derivatives, or oxo; R ¹⁰ is (C 1-6 alkyl unsubstituted or substituted with one or two residues selected from the group consisting of amino, chloro, bromo, fluoro, hydroxy and methoxy or a protected derivative thereof, (C 3-7) cycloalkyl, (C 3-7) cycloalkyl (C 1-5) alkyl or phenyl or phenyl (C 1-4) alkyl unsubstituted or substituted on the phenyl ring by one or two radicals from the group consisting of amino, chloro, bromo, fluoro, hydroxy, methoxy and halogen-unsubstituted or substituted methyl or a protected derivative thereof; R ⁿ is independently (C | -s) alkyl, (C3 "C7) cycloalkyl, (C3-7) cycloalkyl (Ci-s) alkyl or phenyl, and phenyl (C) _6) alkyl unsubstituted or substituted on the phenyl ring with one or two residues selected from the group consisting of amino, chloro, bromo, fluoro, hydroxy, methyl, trifluoromethyl and methoxy; and R ¹² and R ¹³ are (C 1-7) alkyl, (C 3-7) cycloalkyl, (C 3-7) cycloalkyl (C 1-8) alkyl or phenyl and phenyl (C 1-4) alkyl unsubstituted or substituted on the phenyl ring by one or two radicals from the group which consists of amino, chloro, bromo, fluoro, hydroxy, and methoxy and halogen unsubstituted or substituted methyl. The compound of claim 16, wherein n is 0 to 2; Z is - (CH 2 - or -C (R ⁶ ) (R ⁷ ) - and when n is 0, Z is not - (C ¹ H -; R ¹ is hydrogen, (C 1-6) alkoxycarbonyl, (C 2-6) alkanoyl unsubstituted or substituted carboxy rim (Ci_5) alkyloxycarbonyl and hetero (C4-8) cycloalkyl (C4-6) alkanoylamino, -C (O) NR ²² R ^{2 I} wherein R ²¹ together with R ²² form aza _(2-C,) methylene, oxa (C 2-6) methylene or ((n-methylene, (C 3-6 cycloalkylcarbonyl, benzyloxycarbonyl, acetyl, benzoyl or dimethylaminosulfonyl); R ⁸ and R ⁷ are independently (C 5-6) cycloalkyl, (C 5-6 cycloalkylmethyl); 3-pyridyl, 2-thienyl, 2-furyl, 4-imidazolyl, 3-indolyl, 3-pyridylmethyl, 2-thienylmethyl, 2-furylmethyl, 4-imidazolylmethyl, 3-indolylmethyl, methoxy, acetoxy and (C 1-6) alkyl unsubstituted or substituted mercapto, carboxy, amino, methylthio, methylsulfonyl, carbamoyl, dimethylcarbamoyl, guanidino and hydroxy or a protected derivative thereof, phenyl, 1-naphthyl, 2-naphthyl, benzyl, 1-naphthylmethyl, 2-naphthylmethyl and 2-phenylethyl unsubstituted or substituted on the aryl ring with hydroxyl, amino, chlorine, bromine and fluorine or a protected derivative thereof, or together with adjacent R ³ or R ⁵ forms a bivalent (C ^5-6 methylene and 1,2-phenylenedimethylene unsubstituted or substituted with hydroxyl or a protected derivative or oxo group thereof; R ¹⁰ is ethyl, (C 5-6 cycloalkyl, (C 6-6 cycloalkylmethyl or phenyl) and benzyl unsubstituted or substituted on the phenyl ring with amino, hydroxyl, chlorine, bromine or fluorine or a protected derivative thereof; R ¹¹ is ethyl, cyclo (C 5-6) alkyl , cyclo (C _5-6 ) alkylmethyl or phenyl and benzyl unsubstituted or substituted on the phenyl ring with an amino, hydroxyl, chlorine, bromine or fluorine or a protected derivative thereof, and R ¹² and R ¹³ are ethyl, (C 5-6) cycloalkyl, (C 5-6) (4) cycloalkylmethyl or phenyl and benzyl unsubstituted or substituted on the phenyl ring with an amino, hydroxyl, chlorine, bromine or fluorine or a protected derivative thereof. The compound of claim 17, wherein n is 0 or 1; Z is -C (R ⁶ ) (R ⁷ ) -; R ¹ is hydrogen, tert-butoxycarbonyl, benzyloxycarbonyl, acetyl, 3-carboxypropionyl, 3-methoxycarbonylpropionyl, biotinylaminohexanoyl, phenylacetyl, benzoyl, dimethylaminosulfonyl, benzylsulfonyl, 1-piperizinylcarbonyl, 4-methyl-1-piperazinylcarbonyl or 4-morpholinylcarbonyl; R ⁸ is butyl, 2-phenylethyl, 2-methylsulfonylethyl, 2-tert-butoxycarbonylethyl, 2-tert-butoxycarbonylmethyl, 4-tert-butoxycarbonylaminobutyl, 4-benzoylaminobutyl or benzyloxymethyl; and R is 3-pyridylmethyl, 2-thienylmethyl, 2-phenylmethyl, 4-imidazolylmethyl, 3-indolylmethyl and (C 1-8 alkyl unsubstituted or substituted by mercapto, carboxyl, amino, methylthio, methylsulfonyl, carbamoyl, hydroxy, guanamido or dimethylcarbamoyl, or their carbamoyl; a derivative, benzyl, 1-naphthylmethyl, 2-naphthylmethyl and 2-phenylethyl unsubstituted or substituted on the aryl ring with hydroxyl, amino, chlorine, bromine and fluorine or a protected derivative thereof, or together with adjacent R or R forms a divalent radical which is (C 5-6 methylene and 1,2-phenylenedimethylene unsubstituted or substituted by hydroxyl or a protected derivative or oxo group thereof). A compound according to claim 18, wherein n is 0, R ³ , R ⁵ and R ⁶ are hydrogen, R ¹ is hydrogen, tert-butyloxycarbonyl, benzyloxycarbonyl, biotinylaminohexanoyl, benzoyl, 1-piperisinylcarbonyl, 4-methyl-1-piperazinylcarbonyl or 4-morpholinylcarbonyl; R ⁸ is butyl, 2-phenylethyl or 2-methylsulfonylethyl; and R ⁷ is (C 1-6) alkyl, 2-methylsulfonylethyl, unsubstituted or substituted benzyl, 1-naphthylmethyl, 2-naphthylmethyl, 3-pyridinylmethyl or 2-methylsulfonylethyl. The compound of claim 19, wherein R ¹ is 1-piperisinylcarbonyl, 4-methyl-1-piperazinylcarbonyl or 4-morpholinylcarbonyl; R is 2-phenylethyl; and R is unsubstituted or substituted benzyl, 1-naphthylmethyl or 2-naphthylmethyl. A compound according to claim 20, wherein R ¹ is 4-morpholinylcarbonyl, R ⁸ is 2-phenylethyl, R ⁷ is 2-naphthyl-3-methyl and R ⁹ is ethoxycarbonyl which is ethyl 4S- [N- (4-morpholinylcarbonyl) p- (2-naphthyl) -L-alanylamino] -6-phenylhexanoate. A compound according to claim 20, wherein R ¹ is 4-morpholinylcarbonyl, R ⁸ is 2-phenylethyl, R ⁷ is benzyl and R ⁹ is ethoxycarbonyl which is ethyl-4S- [N- (4-morpholinylcarbonyl) L-phenylatanylamino] -6-phenylhexanoate. A compound according to claim 20, wherein R is 4-morpholinylcarbonyl, R is 2-phenylethyl, R is benzyl and R ⁹ is phenylcarbamoyl which is N ² - (4-morpholinylcarbonyl) -N ^1- [3-phenyl-1S ( 2-phenylcarbamoylethyl) propyl] -L-phenylalaninamide. The compound of claim 20, wherein R ¹ is 4-morpholinylcarbonyl, R ⁸ is 2-phenylethyl, R ⁷ is benzyl and R ⁹ is benzylcarbamoyl which is N ² -4- (morpholinylcarbonyl) -N ¹ - [3-phenyl -1S (2-benzylcarbamoylethyl) propyl] -L-phenylalaninamide. • · · · · · 25. A compound of formula (III) fy / fy ' III wherein n is 0 to 13; AB is a linker of the group -C (O) NR ³ -, -CH ² NR ³ -, -C (O) Cl 12 - and -NR ³ C (O) -, wherein R ³ is hydrogen or a group defined below; Y is -CH (R ⁵ ) - or -NR ⁵ -, wherein R ⁵ is hydrogen or as defined below; Z is - (CH 2) 2 -, -C (R ⁶ ) (R ⁷ ) - or -N (R ⁷ ) -, wherein R ⁶ is hydrogen or methyl and R ⁷ is defined as mze; Z ¹ is - (CH ₂ ) ₂ -, -C (R ⁶ ) (R ⁸ ) - or -N (R ⁸ ) -, wherein R ⁶ is hydrogen or methyl and R ⁸ is defined below; R ¹ is hydrogen, alkyloxycarbonylalkanoyl, alkyloxycarbonyl, alkanoyl unsubstituted or substituted by a group consisting of carboxyl, alkyloxycarbonyl and heterocycloalkylalkanoylamino, cycloalkylcarbonyl, heterocycloalkylcarbonyl unsubstituted or substituted by alkylcarbonyl, hydroxy, alkyl, alkoxy and alkoxy, alkyl, alkanoyl, , dialkylcarbamoyl, arylcarbamoyl, arylalkylcarbamoyl, arylalkanoyl, aryloyl, alkylsulfonyl, dialkylaminosulfonyl, arylsulfonyl or heteroarylsulfonyl; R ⁷ and R ⁸ are independently hydrogen, alkyl unsubstituted or substituted by hydroxyl, amino, alkylamino, dialkylamino, uriedo, mercapto, alkylthio, carboxy, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, alkylsulfonyl and guanidine or protected derivatives thereof, cycloalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, an aryl and arylalkyl group unsubstituted or substituted on the aryl ring with one to three radicals selected from the group consisting of: Containing hydroxyl, amino, guanidino, halogen, halogen unsubstituted or substituted alkyl, alkyloxy and aryl groups or their protected derivatives, or together with adjacent R ³ or R ⁵ groups form a divalent radical from the group consisting of (C 1-4) methylene and 1,2-phenylenedimethylene unsubstituted or substituted by hydroxy or protected by hydroxy or oxo; R ¹⁵ is hydrogen, methyl, fluoro or a group of formula (a) or (b) wherein n, A, Β, Y, Z and R ^J are as defined above and R ¹⁰ is hydrogen, alkyl unsubstituted or substituted with one or more radicals selected from from the group consisting of amino, halogen, hydroxyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl or a protected derivative thereof, cycloalkyl, cycloalkylalkyl or aryl and arylalkyl unsubstituted or substituted on the aromatic ring by one or two groups selected from the group consisting of amino, halogen, hydroxyl, halogen unsubstituted or substituted alkyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl or a protected derivative thereof; and R ¹⁶ is phenyl or (C 5-6) heteroaryl unsubstituted or substituted by at least one group such as alkylcarbamoyl, dialkylcarbamoyl, alkyloxycarbonyl, alkylsulfinamoyl, dialkylsulfinamoyl, alkylsulfonyl, carboxyl, nitro, sulfinamoyl, sulfosloxy, carbamoyl, phosphoxyl, carbamoyl, alkylsulfinyl, sulfamoyl, alkylsulfamoyl, dialkylsulfamoyl, alkyloxysulfonyl, alkylsulfonimidoyl, aryl, heteroaryl, hydroxyl, alkyloxy, halogen unsubstituted or substituted alkyl, arylalkyl, halogen, ⁺ N (R ¹⁷ ) 3, wherein R ¹⁷ is independently alkyl, aryl or arylalkyl, or -N (R ¹⁸ ) ₂ wherein R ¹⁸ is independently hydrogen, alkyl, aryl or arylalkyl; and pharmaceutically acceptable salts thereof; individual isomers and mixtures thereof. The compound of claim 25, wherein n is 0 to 5; AB is a linker of the formula -C (O) NR ³ -, Y is -N (R ⁵ ) -, Z is - (CH 2) 2 - or -C (R ⁶ ) (R ⁷ ) -; Z ¹ is -CH (R ⁸ ) -; R ¹ is hydrogen, (C 3 -C 10) alkyloxycarbonylalkanoyl, (C 1-9) alkoxycarbonyl, (C 1-10) alkanoyl unsubstituted or substituted by one of the group consisting of carboxy, (C 1-8) alkyloxycarbonyl and hetero (C 4-6) cycloalkyl _(C2 _io) alkanoylamino, (C ^ jcykloalkylkarbonyl, hetero (C4_g) cycloalkylcarbonyl unsubstituted or substituted with one radical selected from the group consisting of hydroxyl, (C ₅₎ alkyl, (C ₅₎ alkanoyl, (C |. ₅₎ alkyloxycarbonyl , (C 6-10) aryl (C 1-5) alkyloxycarbonyl and hetero (C 4-8) cycloalkylcarbonyl), (C 6-10) alkyl (C 1-5) alkyloxycarbonyl, carbamoyl, (C 1-5 alkylcarbamoyl, di (C 1-5) alkylcarbamoyl, (C _6-10 ) alkyl (C _6-10 ) alkyloxycarbonyl). io) arylcarbamoyl, (C6-io) aryl (Ci-5) alkylcarbamoyl, (C ₆ .io) aryl (Ci-5) alkanoyl, (C ₇ _n) aroyl, (Ci. ₅₎ alkylsulfonyl, di (C-5 ) alkylaminosulfonyl, (C ₆ .io) arylsulfonyl or hetero (C5_8) arylsulfonyI; and R ⁷ and R ⁸ are independently (C _3-7 ) cycloalkyl, (C _3-7 ) cycloalkyl (C 1-5) alkyl, pyridyl, thienyl, furyl, imidazolyl, indolyl, pyridyl (C 1-6) alkyl, thienyl (C 1-6) alkyl, fiiryl (C ^) alkyl, imidazolyl (C-C6) alkyl, indolyl (C ^) alkyl, (C | _ ₅₎ alkyl, _(C2 ^) alkyloxy and (C "5) alkanoyloxy unsubstituted or substituted by a radical from a series consisting of mercapto, carboxy, amino, methylthio, methylsulfonyl, carbamoyl, dimethylcarbamoyl, guanidino and hydroxy or protected derivatives thereof, phenyl, naphthyl, phenyl (C 1-6) alkyl, naphthyl (C 1-6) alkyl, unsubstituted or substituted in the aryl moiety one to three consecutive substituents from the group consisting of hydroxyl, amino, chloro, bromo, fluoro, methyl, trifluoromethyl, methoxy and phenyl, or a protected derivative thereof, or together with adjacent R or R groups form one of the bivalent radicals including (C 6 -methylene and 1,2-phenylenedimethylene unsubstituted or substituted by the following groups from the group consisting of hydroxy or a protected derivative or oxo group; R ¹⁰ is (C 1-8) alkyl unsubstituted or substituted by one or two radicals from the series consisting of amino, chloro, bromo, fluoro, hydroxyl and methoxy, or a protected derivative thereof, (C _1-7 ) cycloalkyl, (C _3-7 ) cycloalkyl (C 1-7) "5) alkyl or phenyl or phenyl (C _1-6 ) alkyl unsubstituted or substituted on the phenyl ring with one or two radicals from the series consisting of amino, chloro, bromo, fluoro, hydroxyl, methoxy and halogen unsubstituted or substituted methyl or a protected derivative thereof; and R ¹⁶ is 2-furyl, 2-thienyl, 2-pyrrololyl, 2-phospholyl, 2-arsolyl, 3-pyridyl or 3-phosphinyl unsubstituted or substituted with at least one residue of the series consisting of (C 1-5) alkylcarbamoyl, difC 1-6 alkylcarbamoyl , (C 1-5) alkyl oxycarbonyl, (C 1-5) alkylsulfinamoyl, di (C 1-5) alkylsulfinamoyl, (C 1-5 alkylsulfonyl, carboxyl, nitro, sulfinamoyl, sulfo, carbamoyl, phosphono, (C 1-5) alkyloxyphosphinyl, di (C 1-5) -5) alkyloxyphosphinyl, (C 1-5) alkanoyl, cyano, (C 1-5) alkylsulfinyl, sulfamoyl, (C 1-5) alkylsulfamoyl, di (C 1-5) alkylsulfamoyl, (C 1-5) alkyloxy-sulfonyl, (C 1-5) alkylsulfonimidoyl, phenyl, naphthyl, pyridyl, thienyl, furyl, imidazolyl, indolyl, hydroxyl, (C 1-8) alkyloxy, halogen unsubstituted or substituted (C 1-8) alkyl, benzyl, halogen, * N (R ⁷ ) 3, wherein R ¹⁷ is independently (C 1-5) alkyl , phenyl or benzyl or -N (R ¹⁸ ) 2, wherein R ¹⁸ is independently hydrogen, (C 1-8) alkyl, phenyl or benzyl. The compound of claim 26, wherein n is 0 to 2; Z is - (CH 2) 2 - or -C (R ⁶ ) (R ⁷ ), except when n is 0, Z is not - (CH 2 h-); R ¹ is hydrogen, (C 4-8) alkoxycarbonyl, (C 2-6) alkanoyl unsubstituted or substituted by some of the group consisting of carboxyl, (C 1-8) alkyloxycarbonyl and hetero (C 4-8) cycloalkyl (C 4-6) alkanoylamino, -C (O) NR ²¹ R ²² wherein R ²¹ and R ^{22 taken} together form aza (C 2-6) methylene, oxa (C 2-6) me (ylene or (C 3-7) methylene, (C 4-8) cycloalkylcarbonyl, benzyloxycarbonyl, acetyl, benzoyl or dimethylaminosulfonyl; and R ⁸ and R ⁷ are independently (C 1-6 cycloalkyl, (C _5-6 ) cycloalkylmethyl, 3-pyridyl, 2-thienyl, 2-furyl, 4-imidazolyl, 3-indolyl, 3-pyridylmethyl, 2-thienylmethyl, 2- turylmethyl, 4-imidazolylmethyl, 3-indolylmethyl, methoxy, acetoxyl, (C 1-6) alkyl unsubstituted or substituted by any of the group consisting of mercapto, carboxy, amino, methylthio, methylsulfonyl, carbamoyl, dimethylcarbamoyl, guanidino and hydroxy or their hydroxy groups derivatives, phenyl, 1-naphthyl, 2-naphthyl, benzyl, 1-naphthylmethyl, 2-naphthylmethyl and 2-phenylethyl on the aromatic ring unsubstituted or substituted by a radical selected from the group consisting of hydroxy, amino, chloro, bromo and fluoro or their protected radicals, or together with adjacent R ³ or R ⁵ groups of the following bivalent residues (Cj ^ jmetylenlenlen··· C· C C·) * and 1,2-phenylenedimethylene unsubstituted or substituted by hydroxy or its protected derivatives, or oxo; R ¹⁰ is ethyl, (C _5-6 ) cycloalkyl, (C _5-6 ) cycloalkylmethyl or phenyl and benzyl unsubstituted or substituted on the phenyl ring with amino, hydroxyl, chlorine, bromine or fluorine or a protected derivative thereof; and R ¹⁶ is 2-furyl, 2-thienyl, 2-pyrrolyl, 2-phospholyl, 2-arsolyl, 2-pyridyl or 3-phosphinyl unsubstituted or substituted by at least one of methylcarbamoyl, dimethylcarbamoyl, methyloxycarbonyl, methylsulfinamoyl, dimethylsulfinamoyl, methylsulfonyl, carboxyl, nitro, sulfinamoyl, sulfo, carbamoyl, phosphono, methyloxyphosphinyl, dimethyloxyphosphinyl, formyl, cyano, methylsulfinyl, sulfamoyl, methylsulfamoyl, dimethylsulfamoyl, phenylsulfonyl, phenylsulfonyl, phenylsulfonyl, phenylsulfonyl, phenylsulfonyl, phenylsulfonyl; hydroxyl, methoxy, methyl, trifluromethyl, benzyl, halo, N (R ⁱ⁷⁾ 3 where R ¹⁷ is independently methyl, phenyl or benzyl, or -N (R ⁱ⁸⁾ 2 wherein R ¹⁸ is independently hydrogen, methyl, phenyl or benzyl. The compound of claim 27 wherein n is 0 to 1; Z is -C (R ⁶ ) (R ⁷ ) -; R ¹ is hydrogen, tert-butoxycarbonyl, benzyloxycarbonyl, acetyl, 3-carboxypropionyl, 3-methoxycarbonylpropionyl, biotinylaminohexanoyl, phenylacetyl, benzoyl, dimethylaminosulfonyl, benzylsulfonyl, 1-piperizinylcarbonyl, 4-methyl-1-piperazinylcarbonyl or 4-morpholinylcarbonyl; R ⁸ is butyl, 2-phenylethyl, 2-methylsulfonylethyl, 2-tert-butoxycarbonylethyl, 2-tert-butoxycarbonylmethyl, 4-tert-butoxycarbonylaminobutyl, 4-benzoylaminobutyl or benzyloxymethyl; and R ⁷ is 3-pyridylmethyl, 2-thienylmethyl, 2-furylmethyl, 4-imidazolylmethyl, 3-indolylmethyl, (C 1-5) alkyl unsubstituted or substituted by one of mercapto, carboxy, amino, methylthio, methylsulfonyl, carbamoyl, dimethylcarbamoyl, guanidino and hydroxy, or a protected derivative thereof, benzyl, 1-naphthylmethyl, 2- naphthylmethyl and 2-phenylethyl on the aryl ring unsubstituted or substituted by a radical selected from the group consisting of hydroxy, amino, chlorine, bromine and fluorine or a protected derivative thereof, or together with adjacent groups R ³ or R ⁵ form one of the following bivalent radicals jmetylen ·· ················· 9 9 · 9 And 1,2-phenylenedimethylene unsubstituted or substituted by a hydroxy group, or a protected derivative thereof, or an oxo group. The compound of claim 28, wherein n is 0; R ³ , R ⁵ and R ⁶ are hydrogen; R ¹ is hydrogen, tert-butoxycarbonyl, benzyloxycarbonyl, biotinylaminohexanoyl, benzoyl, 1-piperizinylcarbonyl, 4-methyl-1-piperazinylcarbonyl or 4-morpholinylcarbonyl; R is butyl, 2-phenylethyl or 2-methylsultonylethyl; and R ⁷ is (C 1-6 alkyl, 2-methylsulfonylethyl, unsubstituted or substituted benzyl, 1-naphthylmethyl, 2-naphthylmethyl, 3-pyridinylmethyl or 2-methylsulfonylethyl. The compound of claim 29, wherein R ¹ is 1-piperisinylcarbonyl, 4-methyl-1-piperazinylcarbonyl or 4-morpholinylcarbonyl; R is 2-phenylethyl; and R is unsubstituted or substituted benzyl, 1-naphthylmethyl or 2-naphthylmethyl. A compound according to claim 30, wherein R ¹ is 4-morpholinylcarbonyl, R ⁸ is 2-phenylethyl, R ⁷ is benzyl, R ¹⁵ is hydrogen and R ¹⁶ is 4-methoxyphenyl which is N ₂ -4-morpholinylcarbonylN ¹ - { 3-phenyl-1 S- [2- (4-methoxyphenyl) ethyl] propyl} -L-phenylalaninamide. The compound of claim 30, wherein R ¹ is 4-morpholinylcarbonyl, R ⁸ is 2-phenylethyl, R ⁷ is benzyl, R ¹⁵ is hydrogen and R ¹⁶ is 4-aminophenyl, which is N ² - (4-morpholinylcarbonyl) N ^1- {3-Phenyl-1 S- [2- (4-aminophenyl) ethyl] propyl} -L-phenylalaninamide. 33. A method of inhibiting an eysteine protease comprising recovering an inhibitor to an eysteine protease, wherein said inhibitor is an eysteine protease inhibitor according to claim 1. 34. A method of inhibiting an eysteine protease comprising recovering an inhibitor to an eysteine protease, wherein said inhibitor is an eysteine protease inhibitor according to claim 2. 35. A method of inhibiting an eysteine protease comprising recovering an inhibitor to an eysteine protease, wherein said inhibitor is an eysteine protease inhibitor according to claim 3. ·· ··· ♦ 86.:. .:. 36. A method of inhibiting a cysteine protease comprising recovering the inhibitor to a cysteine protease, wherein said inhibitor is a cysteine protease inhibitor according to claim 14. 37. A method of inhibiting a cysteine protease comprising recovering the inhibitor to a cysteine protease, wherein said inhibitor is a cysteine protease inhibitor according to claim 24. 38. A method of treating a patient condition capable of ameliorating the inhibition of a cysteine protease comprising administering a therapeutically effective amount of a cysteine protease inhibitor according to claim 1. 39. A method of treating a patient condition capable of ameliorating the inhibition of a cysteine protease comprising administering a therapeutically effective amount of a cysteine protease inhibitor according to claim 2. 40. A method of treating a patient condition capable of ameliorating the inhibition of a cysteine protease comprising administering a therapeutically effective amount of a cysteine protease inhibitor according to claim 3. 41. A method of treating a patient condition capable of ameliorating the inhibition of a cysteine protease comprising administering a therapeutically effective amount of a cysteine protease inhibitor according to claim 14. 42. A method of treating a patient condition capable of ameliorating the inhibition of a cysteine protease comprising administering a therapeutically effective amount of a cysteine protease inhibitor according to claim 24. 43. A pharmaceutical composition comprising a therapeutically effective amount of a cysteine protease inhibitor according to claim 1, one of its isomers, a mixture of isomers, or a pharmaceutically acceptable salt thereof, in combination with one or more administration agents. 44. A pharmaceutical composition comprising a therapeutically effective amount of a cysteine protease inhibitor according to claim 2, a single isomer, a mixture of isomers, or a pharmaceutically acceptable salt thereof, in combination with one or more administration agents. 45. A pharmaceutical composition comprising a therapeutically effective amount of a cysteine protease inhibitor according to claim 3, a single isomer, a mixture of isomers, or a pharmaceutically acceptable salt thereof in combination with one or more administration agents. 46. A pharmaceutical composition comprising a therapeutically effective amount of a cysteine protease inhibitor according to claim 14, a single isomer, a mixture of isomers, or a pharmaceutically acceptable salt thereof, in combination with one or more administration agents. 47. A pharmaceutical composition comprising a therapeutically effective amount of a cysteine protease inhibitor according to claim 24, a single isomer, a mixture of isomers, or a pharmaceutically acceptable salt thereof in combination with one or more administration agents. A process for preparing a compound of formula IV o R H IV wherein n is 0 to 12; AB is a linker selected from -C (O) NR ³ -, -CH ² NR ³ -, -C (O) CH 2 - and -NR ³ C (O) -, wherein R ³ is hydrogen or a group defined below; Y is -CH (R ⁵ ) - or -NR ⁵ -, wherein R ⁵ is hydrogen or as defined below; Z is - (CH ₂ ) ₂ -, -C (R ⁶ ) (R ⁷ ) - or -N (R ⁷ ) -, wherein R ⁶ is hydrogen or methyl and R ⁷ is defined below; 9 999 Z ¹ is - (CH ₂ ) ₂ -, -C (R ⁶ ) (R ⁸ ) - or -N (R ⁸ ) -, wherein R ⁶ is hydrogen or methyl and R ⁸ is A 7 can be defined; R is hydrogen or methyl and R is defined below; R ¹ is hydrogen, alkyloxycarbonylalkanoyl, alkyloxycarbonyl, alkanoyl unsubstituted or substituted by a radical selected from the group consisting of carboxyl, alkyloxycarbonyl and heterocycloalkylalkanoylamino, cycloalkylcarbonyl, heterocycloalkyloxycarbonyl, heterocycloalkyloxyalkyl, alkoxyalkyloxy, alkyl, alkoxyalkylcarbonyl, alkyl, alkoxyalkyl, dialkylcarbamoyl, arylcarbamoyl, arylalkylcarbamoyl, arylalkanoyl, aryloyl, alkylsulfonyl, dialkylaminosulfonyl, aylsulfonyl or heteroarylsulfonyl; R and R are independently hydrogen, alkyl unsubstituted or substituted by hydroxyl, amino, alkylamino, dialkylamino, uriedo, mercapto, alkylthio, carboxy, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, alkylsulfonyl and guanidine or protected derivatives thereof, cycloalkyl, cycloalkylalkyl heteroaryl, heteroarylalkyl, an aryl and arylalkyl group unsubstituted or substituted on the aryl ring with one to three radicals selected from the group consisting of hydroxyl, amino, guanidino, halogen, halogen unsubstituted or substituted alkyl, alkyloxy and aryl groups or protected derivatives thereof; or together with adjacent R or R groups forms a divalent radical (C 6 / methylene and 1,2-phenylenedimethylene unsubstituted or substituted by hydroxy or protected by hydroxy or oxo; and R ²⁰ is cyano, -S (O) 2 R ² , -CH ² S (O) ₂ R ² , -CH ² CH (R ⁴ ) S (O) 2 R ² , - (CH ₂ ) ₂ C (O) OR ^{1 ()} , - (CH 2) 2 P (O) (OR ^{, O} ) 2, - (CH ₂ ) ₂ S (O) (NR ¹⁰ ) R ¹⁰ , - ( CH 2) 2 C (O) R ^h , - (CH 2) 2 S (O) R ¹ ', - (CH 2) 2 C (O) NR' ² R ¹³ , - (CH 2) 2 S (O) ₂ NR ¹² R ' ³ , - (CH 2) 2 C (O) NHR ¹⁴ , - (CH 2) ₂ S (O) ₂ NHR ¹⁴ or -CH ₂ CHR ¹⁵ R ¹⁶ , wherein R ² is hydrogen, alkyl unsubstituted or substituted by one or more radicals from the group consisting of amino, halogen, hydroxy, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl or protected derivatives thereof, cycloalkyl, cycloalkylalkyl or aryl and arylalkyl groups unsubstituted or substituted on the aryl ring with one to two radicals selected from the group consisting of amino, halogen, hydroxyl, halogen 4 4 Unsubstituted or substituted alkyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl or protected derivatives thereof; R ⁴ is hydrogen, alkyl or arylalkyl, R ¹⁰ is independently hydrogen, alkyl unsubstituted or substituted by one or more of the series consisting of amino, halogen, hydroxyl, alkyloxy, nitro, alkylsulfonyl and aylsulfonyl or a protected derivative thereof, cycloalkyl, cycloalkylalkyl or aryl and arylalkyl unsubstituted or substituted on the aryl ring one or two radicals from the series comprising amino, halogen, hydroxy, optionally halo-substituted alkyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl, or a protected derivative thereof, R ⁿ is hydrogen, alkyl, perfluoroalkyl, cycloalkyl, cycloalkylalkyl, perfluoroaryl, perfluoraiylakyl or aryl and arylalkyl unsubstituted or substituted on the aryl ring with a series of amino, halogen, hydroxyl, halogen unsubstituted or substituted alkyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl or a protected derivative thereof, R ¹² and R ¹³ are independently hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl or arylalkyl, R ¹⁴ is -C (O) OR ¹⁰ , where R ¹⁰ is as defined above or a group of formulas (a) and (b) (a) Φ) wherein n, A, Β, Y, Z, R ¹ and R ¹⁰ are as defined above, ^{R a} is hydrogen, methyl, fluoro or a group of formula (a) or (b) above, and R ¹⁶ is phenyl or (C ₅ .6) heteroaryl unsubstituted or substituted with at least one group from the series consisting alkylcarbamoyl , dialkylcarbamoyl, alkoxycarbonyl, alkylsulfinamoyl, dialkylsulfmamoyl, alkylsulfonyl, carboxyl, nitro, sulfinamoyl, sulfo, carbamoyl, phosphono, alkyloxyphosphinyl, dialkyloxyphosphinyl, alkanoyl, cyano dial, alkylsulfamyl, alkylsulfamyl, sulfamylsulfamyl, sulfamylsulfamyl; 4 44 4444 4 * 44 4 4 4 4 4,444 444 4 4 44 • 44 444 44 4 44 ·· 4 4 4 4 44 4 4 4 4 4 4444 44 alkyloxysulfonyl, alkylsulfonimidoyl, aryl, heteroaryl, hydroxyl, alkyloxy, halogen substituted or unsubstituted alkyl, arylalkyl, halogen, ⁺ N (R ¹⁷ ) 3, wherein R ¹⁷ is independently alkyl, aryl or arylalkyl or -N (R ¹⁸ ) 2 wherein R ¹⁸ is independently hydrogen, alkyl, aryl or arylalkyl; and pharmaceutically acceptable salts, individual isomers, and mixtures thereof, comprising (A) reacting an amine of formula V R * H iN with a compound of formula VI o VI wherein n, A, Β, X, Y, Z, R ¹ , R ⁸ and R ²⁰ are as defined above; (B) preferably converting the free form of a compound of Formula IV into a pharmaceutically acceptable salt; (C) preferably converting a salt of a compound of Formula IV into the free form; (D) preferably separating the individual stereoisomers of the compound of formula IV. A process for preparing a compound of formula IV H IV wherein n is 0-12; n is 0 to 12; AB is a linker of the group -C (O) NR ³ -, -CH ² NR ³ -, -C (O) CH 2 - and -NR ³ C (O) -, wherein R ³ is hydrogen or a group defined below; Y is -CH (R ⁵ ) - or -N (R ⁵ ) - wherein R ⁵ is hydrogen or as defined below; Z is - (CH ₂ ) ₂ -, -C (R ⁶ ) (R ⁷ ) - or -N (R ⁷ ) -, wherein R ⁶ is hydrogen or methyl and R ⁷ is as defined herein; Z ¹ is - (CH ₂ ) 2 -, -C (R ⁶ ) (R ⁸ ) - or -N (R ⁸ ) -, wherein R ⁶ is hydrogen or methyl and R ⁸ is as defined below; R ⁶ is hydrogen or methyl and R ⁷ is defined below; R ¹ is hydrogen, alkyloxycarbonylalkanoyl, alkyloxycarbonyl, alkanoyl unsubstituted or substituted by a group consisting of carboxyl, alkyloxycarbonyl and heterocycloalkylalkanoylamino, cycloalkylcarbonyl, heterocycloalkylcarbonyl unsubstituted or substituted by carbocycloalkyl, alkyl, hydroxy, alkyl, hydroxy, alkyl, hydroxy, alkyl, hydroxy, alkyl, , alkylcarbamoyl, dialkylcarbamoyl, arylcarbamoyl, arylalkylcarbamoyl, arylalkanoyl, aryloyl, alkylsulfonyl, dialkylaminosulfonyl, arylsulfonyl or heteroarylsulfonyl; R and R are independently hydrogen, alkyl unsubstituted or substituted with hydroxyl, amino, alkylamino, dialkylamino, uriedo, mercapto, alkylthio, carboxy, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, alkylsulfonyl and guanidino or a protected derivative thereof, cycloalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, aryl and arylalkyl unsubstituted or substituted on the aryl ring by one to three radicals selected from the group consisting of hydroxyl, amino, halogen, halogen-unsubstituted or substituted alkyl, alkyloxy and aryl groups or protected derivatives thereof, or together with adjacent R or R groups is a divalent radical from the group consisting of (C 1-4) methylene and 1,2-phenylenedimethylene unsubstituted or substituted by hydroxy or protected by hydroxy or oxo; and R is -S (O) 2 R, wherein R is hydrogen, alkyl unsubstituted or substituted with one or more amino, halogen, hydroxyl, alkyloxy, radicals; Nitro, alkylsulfonyl and arylsulfonyl or protected derivatives thereof, cycloalkyl, cycloalkylalkyl, aryl and arylalkyl unsubstituted or substituted on the aryl ring by one or two residues of the series consisting of amino , halogen, hydroxyl, halogen-unsubstituted or substituted alkyl, alkyloxy, nitroalkylsulfonyl and arylsulfonyl or protected derivatives thereof, or pharmaceutically acceptable salts thereof, individual isomers and mixtures of isomers, comprising (A) reacting a compound of formula (VII): With an aldehyde of the formula R ⁸ CHO and a sodium sulfinate of the formula R ² S (O) ONa, wherein n, A, Β, X, Y, Z, R ¹ and R ⁸ are as defined above; (B) preferably converting the free form of a compound of Formula IV into a pharmaceutically acceptable salt; (C) preferably converting a salt of a compound of Formula IV into the free form; (D) preferably separating the individual stereoisomers of the compound of formula IV. 50. A process for preparing a compound of formula IV H IV wherein n is 0 to 12; AB is a linker of the group -C (O) NR ³ -, -CH ² NR ³ -, -C (O) CH 2 - and -NR ³ C (O) -, wherein R ³ is hydrogen or a group defined below; • · Y is -CH (R ⁵ ) - or -N (R ⁵ ) - wherein R ⁵ is hydrogen or as defined below; Z is - (CH ₂ ) ₂ -, -C (R ⁶ ) (R ⁷ ) - or -N (R ⁷ ) -, wherein R ⁶ is hydrogen or methyl and R ⁷ is defined below; Z ¹ is - (CH ₂ ) ₂ -, -C (R ⁶ ) (R ⁸ ) - or -N (R ⁸ ) -, wherein R ⁶ is hydrogen or methyl and R ⁸ is defined below; R ⁶ is hydrogen or methyl and R ⁷ is defined below; R ¹ is hydrogen, alkyloxycarbonylalkanoyl, alkyloxycarbonyl, alkanoyl unsubstituted or substituted by a radical selected from the group consisting of carboxyl, alkyloxycarbonyl and heteroeykloalkylalkanoylamino, cycloalkylcarbonyl, heterocycloalkylcarbonyl unsubstituted or substituted carbocycloalkyl, alkyl, hydroxy, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, arylcarbamoyl, arylalkylcarbamoyl, arylalkanoyl, aryloyl, alkylsulfonyl, dialkylaminosulfonyl, arylsulfonyl or heteroarylsulfonyl; R and R are independently hydrogen, alkyl unsubstituted or substituted by hydroxyl, amino, alkylamino, dialkylamino, uriedo, mercapto, alkylthio, carboxy, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, alkylsulfonyl and guanidine or protected derivatives thereof, cycloalkyl, cycloalkylalkyl heteroaryl, heteroarylalkyl, aryl and arylalkyl unsubstituted or substituted on the aryl ring by one to three radicals selected from the group consisting of hydroxyl, amino, halogen, halogen unsubstituted or substituted alkyl, alkyloxy and aryl groups or protected derivatives thereof, or together with adjacent R or R groups form a divalent radical _(C-3) methylene and 1,2-phenylenedimethylene unsubstituted or substituted by hydroxy or protected hydroxy, or oxo; and R ²⁰ is -S (O) ₂ R ² wherein R ² is hydrogen, alkyl unsubstituted or substituted by one or more of amino, halogen, hydroxyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl or protected derivatives thereof, cycloalkyl, cycloalkylalkyl or aryl and arylalkyl unsubstituted or substituted on the aryl ring by one or two groups of the series consisting of amino, halogen, hydroxyl, halogen-unsubstituted or substituted alkyl, alkoxy, 4 4 4 · · · 44 4 · · · · 4444 44 444 444 44 nitro, alkylsulfonyl and arylsulfonyl or a protected derivative thereof; and pharmaceutically acceptable salts, individual isomers and mixtures thereof, comprising (A) (1) reacting a compound of formula NH ₂ P wherein P is a protecting group with an aldehyde of the formula R ⁸ CHO and a sodium sulfinate of formula R ² S (O) ONa and subsequent deprotection to give a compound of formula VIII VIII, wherein R ² and R ⁸ are as defined above; and (2) reacting a compound of Formula VIII with a compound of Formula VIo ./K _{b 2} ^z '^ b / ^a x ^ K2A. _oh VI where η, A, Β, X, Y, Z and R ¹ are as defined above; (B) preferably converting the free form of a compound of Formula IV into a pharmaceutically acceptable salt; (C) preferably converting a salt of a compound of Formula IV into the free form; (D) preferably separating the individual stereoisomers of the compound of formula IV. Process for preparing a compound of formula IV IV wherein n is 0 to 12; AB is a linker of the group -C (O) NR ³ -, -CH ² NR ³ -, -C (O) CH 2 - and -NR ³ C (O) -, wherein R ³ is hydrogen or a group defined below; Y is -CH (R ⁵ ) - or -N (R ⁵ ) - wherein R ⁵ is hydrogen or as defined below; Z is - (CH ₂ ) ₂ -, -C (R ⁶ ) (R ⁷ ) - or -N (R ⁷ ) -, wherein R ⁶ is hydrogen or methyl and R ⁷ is as defined herein; Z ¹ is - (CH ₂ ) ₂ -, -C (R ⁶ ) (R ⁸ ) - or -N (R ⁸ ) -, wherein R ⁶ is hydrogen or methyl and R ⁸ is defined below; R is hydrogen or methyl and R is defined below; R ¹ is hydrogen, alkyloxycarbonylalkanoyl, alkyloxycarbonyl, alkanoyl unsubstituted or substituted by a group consisting of carboxyl, alkyloxycarbonyl and heterocycloalkylalkanoylamino, cycloalkylcarbonyl, heterocycloalkylcarbonyl unsubstituted or substituted by carbocycloalkyl, alkyl, hydroxy, alkyl, hydroxy, alkyl, hydroxy, alkyl, hydroxy, alkyl, , alkylcarbamoyl, dialkylcarbamoyl, aylcarbamoyl, arylalkylcarbamoyl, arylalkanoyl, aryloyl, alkylsulfonyl, dialkylaminosulfonyl, arylsulfonyl or heteroarylsulfonyl; R and R are independently hydrogen, alkyl unsubstituted or substituted by hydroxyl, amino, alkylamino, dialkylamino, uriedo, mercapto, alkylthio, carboxy, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, alkylsulfonyl and guanidine or protected derivatives thereof, cycloalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, aryl and arylalkyl unsubstituted or substituted on the aryl ring by one to three radicals selected from the group consisting of hydroxyl, amino, halogen, halogen unsubstituted or substituted alkyl, alkyloxy and aryl groups or protected derivatives thereof, or together with adjacent R or R groups form the divalent radical (C3-1) of methylene and 1,2-phenylenedimethylene. 9 9 φ Φ · · ΦΦΦ · 96: unsubstituted or substituted by hydroxy or protected by hydroxy or oxo; and R ²⁰ is -S (O) 2 R ² , wherein R ² is hydrogen, alkyl unsubstituted or substituted by one or more of amino, halogen, hydroxyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl or protected derivatives thereof, cycloalkyl, cycloalkylalkyl or aryl and alkylalkyl unsubstituted or substituted on the aryl ring by one or two of the series consisting of amino, halogen, hydroxyl, halogen unsubstituted or substituted alkyl, alkoxy, nitro, alkylsulfonyl and arylsulfonyl or a protected derivative thereof; and pharmaceutically acceptable salts, individual isomers and mixtures thereof, comprising (A) (1) reacting a compound of Formula IX With a thiolate anion of the formula R S *, wherein L is a leaving group and R and R are as defined above to form a compound of formula X R * SR ² PHN (2) oxidizing a compound of formula X to give a compound of formula XI S (O) ₂ R ² PHN (3) reacting a compound of formula XI with a compound of formula VI, wherein η, A, Β, X, Y, Z and R ¹ are as defined above; (B) preferably converting the free form of a compound of Formula IV into a pharmaceutically acceptable salt; (C) preferably converting a salt of a compound of Formula IV into the free form; (D) preferably separating the individual stereoisomers of the compound of formula IV. 52. A process for the preparation of a compound of formula IV of R * H IV wherein n is 0 to 12; AB is a linker of the group -C (O) NR ³ -, -CH ² NR ³ -, -C (O) CH 2 - and -NR ³ C (O) -, wherein R ³ is hydrogen or a group defined below; Y is -CH (R ⁵ ) - or -NR ⁵ -, wherein R ⁵ is hydrogen or as defined below; Z is - (O 2) 2 -, -C (R ⁶ ) (R ⁷ ) - or -N (R ⁷ ) -, wherein R ⁶ is hydrogen or methyl and R ⁷ is defined below; Z * is - (O 2) 2 -, -C (R ⁶ ) (R ⁸ ) - or -N (R ⁸ ) -, wherein R ⁶ is hydrogen or methyl and R ⁸ is as defined below; R ⁶ is hydrogen or methyl and R ⁷ is defined below; R @ ¹ is hydrogen, alkyloxycarbonylalkanoyl, alkyloxycarbonyl, alkanoyl unsubstituted or substituted by a radical selected from the group consisting of carboxyl, alkyloxycarbonyl, and -. Heterocycloalkylalkanoylamino, cycloalkylcarbonyl, heterocycloalkylcarbonyl unsubstituted or substituted by a group selected from hydroxyl, alkyl, alkanoyl, alkyloxycarbonyl, arylalkyloxycarbonyl, heterocycloalkylcarboyl, alkylcarbamoyl, arylalkyloxycarbonyl, arylalkyloxycarbonyl, , alkylsulfonyl, dialkylaminosulfonyl, arylsulfonyl or heteroarylsulfonyl; R ⁷ and R ⁸ are independently hydrogen, alkyl unsubstituted or substituted with hydroxyl, amino, alkylamino, dialkylamino, uriedo, mercapto, alkylthio, carboxy, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, alkylsulfonyl and guanidine or protected derivatives thereof, cycloalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, ayl and arylalkyl unsubstituted or substituted on the aryl ring by one to three radicals selected from the group consisting of hydroxyl, amino, halogen, halogen unsubstituted or substituted alkyl, alkyloxy and aryl groups or protected derivatives thereof, or together with adjacent R or R groups form a divalent radical from the group consisting of (C 1-4) methylene and 1,2-phenylenedimethylene unsubstituted or substituted by hydroxy or protected by hydroxy or oxo; and R ²⁰ is cyano, -S (O) 2 R ² , -CH ² S (O) ₂ R ² , -CH ² CH (R ⁴ ) S (O) 2 R ² , - (CH ₂ ) ₂ C (O) QR ' ⁰ , - (CH 2) 2 P (O) (OR ¹⁰ ) 2, - (CH ₂ ) ₂ S (O) (NR ¹⁰ ) R ^{, ()} , - (CH 2) ) 2C (O) R ^h, - (CH 2) ₂ S (O) R ^h, - (CH2) 2C (O) NR ⁱ² R ^13, - (CH 2) ₂ S (O) ₂ NR ^I2 R ¹³ - ( CH 2) 2 C (O) NHR ¹⁴ , - (CH 2) ₂ S (O) ₂ NHR ^14, or -CH ₂ CHR ¹⁵ R ¹⁶ , wherein R ² is hydrogen, alkyl unsubstituted or substituted with one or more amino, halogen , hydroxy, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl or protected derivatives thereof, cycloalkyl, cycloalkylalkyl or aryl and arylalkyl groups unsubstituted or substituted on the aryl ring with one to two radicals selected from the group consisting of amino, halogen, hydroxyl, halogen unsubstituted or substituted alkyl, alkyloxy nitro, alkylsulfonyl and arylsulfonyl or protected derivatives thereof; R ¹⁰ is hydrogen, alkyl unsubstituted or substituted by one or more of the series consisting of amino, halogen, hydroxyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl or a protected derivative thereof, cycloalkyl, cycloalkylalkyl QQ · · ♦ · · · Or aryl and arylalkyl unsubstituted or substituted on the aryl ring by one or two of the series consisting of amino, halogen, hydroxyl, halogen unsubstituted or substituted alkyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl, or a protected derivative thereof, R ⁿ is hydrogen, alkyl, perfluoroalkyl, cycloalkyl, cycloalkylalkyl, perfluoroaryl, perfluorarylakyl or aryl and arylalkyl is unsubstituted or substituted on the aryl ring by one or two radicals from the series comprising amino, halogen, hydroxyl, halogen unsubstituted or substituted alkyl, alkyloxy, nitro, alkylsulfonyl and arylsulfonyl or a protected derivative thereof, R ¹² and R ¹³ are independently hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl or arylalkyl, R ¹⁴ is -C (O) OR ¹⁰ , wherein R ¹⁰ is as defined above or a group of formula (a) or (b) wherein n A, B, Y, Z, R ¹ and R ¹⁰ are as defined above; and pharmaceutically acceptable salts, individual isomers, and mixtures thereof, comprising (A) (1) reacting an aldehyde of formula (XII) PHN R ¹ CHO XII with a compound of the formula selected from ΧΠΙ and XIV 100 o R ²⁰ XIII R XIV wherein R ⁸ and R ²⁰ are as defined above and then deprotected to give a compound of formula XV (2) by reacting a compound of formula XV with a compound of formula VI, o VI where η, A, Β, X, Y, Z and R ¹ are as defined above and (3) a reduction; (B) preferably converting the free form of a compound of Formula IV into a pharmaceutically acceptable salt; (C) preferably converting a salt of a compound of Formula IV into the free form; (D) preferably separating the individual stereoisomers of the compound of formula IV. 53. A process for the preparation of a compound of formula IV of R * TO IV • · · · · · 101.:. .:. wherein n is 0 to 12; AB is a linker selected from -C (O) NR ³ -, -CH ² NR ³ -, -C (O) CH 2 - and -NR ³ C (O) -, wherein R ³ is hydrogen or a group defined below; Y is -CH (R ⁵ ) - or -NR ⁵ -, wherein R ⁵ is hydrogen or as defined below; Z is - (CH ₂ ) 2 -, -C (R ⁶ ) (R ⁷ ) - or -N (R ⁷ ) -, wherein R ⁶ is hydrogen or methyl and R ⁷ is defined below; Z ¹ is - (CH ₂ ) ₂ -, -C (R ⁶ ) (R ⁸ ) - or -N (R ⁸ ) -, wherein R ⁶ is hydrogen or methyl and R ⁸ is defined below; R ⁶ is hydrogen or methyl and R ⁷ is defined below; R ¹ is hydrogen, alkyloxycarbonylalkanoyl, alkyloxycarbonyl, alkanoyl unsubstituted or substituted by a group consisting of carboxyl, alkyloxycarbonyl and heterocycloalkylalkanoylamino, cycloalkylcarbonyl, heterocycloalkylcarbonyl unsubstituted or substituted by alkylcarbonyl, hydroxy, alkyl, alkoxy and alkoxy, alkyl, alkanoyl, , dialkylcarbamoyl, arylcarbamoyl, arylalkylcarbamoyl, aiylalkanoyl, aryloyl, alkylsulfonyl, dialkylaminosulfonyl, arylsulfonyl or heteroarylsulfonyl; R ⁷ and R ⁸ are independently hydrogen, alkyl unsubstituted or substituted by hydroxyl, amino, alkylamino, dialkylamino, uriedo, mercapto, alkylthio, carboxy, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, alkylsulfonyl and guanidine, or protected derivatives thereof, cycloalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, aryl and arylalkyl unsubstituted or substituted on the aryl ring by one to three radicals selected from the group consisting of hydroxyl, amino, halogen, halogen unsubstituted or substituted alkyl, alkyloxy and aryl groups or protected derivatives thereof, or together with adjacent R ³ or the R ⁵ groups form a divalent radical from the group consisting of (C 1-4) methylene and 1,2-phenylenedimethylene unsubstituted or substituted by hydroxy or protected by hydroxy or oxo; and R ²⁰ is -CH ² CHR ¹⁵ R ¹⁶ , wherein R ¹⁵ is hydrogen, methyl, fluoro, or a group of formula (a) or (b). 4 * group of formula (a) or (b) 102 where η, A, Β, Y, Z, R ¹ and R ¹⁰ are as defined above; and R ¹⁶ is phenyl or (C 5-6) heteroaryl unsubstituted or substituted by at least one of alkylcarbamoyl, dialkylcarbamoyl, alkyloxycarbonyl, alkylsulfinamoyl, dialkylsulfinamoyl, alkylsulfonyl, carboxyl, nitro, sulfinamoyl, dialkyl, sulfosinyl, carbamoyl, alkanoyl, cyano, alkylsulfinyl, sulfamoyl, alkylsulfamoyl, dialkylsulfamoyl, alkyloxysulfonyl, alkylsulfonimidoyl, aiyl, heteroaryl, hydroxyl, alkyloxy, halogen unsubstituted or substituted alkyl, arylalkyl, halogen, ⁴ N (R ¹⁷ ) 3, wherein aryl or R ¹⁷ is 3, wherein arylalkyl, or N (R ¹⁸⁾ 2 wherein R ¹⁸ is hydrogen, alkyl, aryl or arylalkyl; and the pharmaceutically acceptable salts, individual isomers and mixtures thereof, characterized in that it comprises (A) (1) the reaction of an aldehyde of formula XII R * XII with a compound of formula XVI XVI 103 wherein R ⁸ , R ¹⁵ and R ¹⁶ are as defined above followed by deprotection to give a compound of formula (XVII) ³ XVII (2) reacting a compound of formula XVII with a compound of formula VI O OH VI wherein n, A, Β, X, Y, Z and R ¹ are as defined above and (3) a reduction; (B) preferably converting the free form of a compound of Formula IV into a pharmaceutically acceptable salt; (C) preferably converting the salt of the compound of formula TV to the free form; (D) preferably separating the individual stereoisomers of the compound of formula IV.