HK1029936B - Quinoline-containing alpha-ketoamide cysteine and serine protease inhibitors - Google Patents

Description

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims benefit of U.S. Provisional Application Serial No. 60/061,267, filed October 7, 1997, and U.S. Application entitled "Quinoline-containing α-ketoamide Cysteine and Serine Protease Inhibitors" filed October 6, 1998.

FIELD OF THE INVENTION

This invention relates to quinoline-containing α-ketoamide inhibitors of cysteine and serine proteases, methods for making these compounds, and methods for using the same.

BACKGROUND OF THE INVENTION

Numerous cysteine and serine proteases have been identified in human tissues. A "protease" is an enzyme which degrades proteins into smaller components (peptides). The terms "cysteine protease" and "serine protease" refer to proteases which are distinguished by the presence therein of a cysteine or serine residue which plays a critical role in the catalytic process. Mammalian systems, including humans, normally degrade and process proteins via a variety of enzymes including cysteine and serine proteases. However, when present at elevated levels or when abnormally activated, cysteine and serine proteases may be involved in pathophysiological processes.

For example, calcium-activated neutral proteases ("calpains") comprise a family of intracellular cysteine proteases which are ubiquitously expressed in mammalian tissues. Two major calpains have been identified; calpain I and calpain II. While calpain II is the predominant form in many tissues, calpain I is thought to be the predominant form in pathological conditions of nerve tissues. The calpain family of cysteine proteases has been implicated in many diseases and disorders, including neurodegeneration, stroke, Alzheimer's, amyotrophy, motor neuron damage, acute central nervous system injury, muscular dystrophy, bone resorption, platelet aggregation, cataracts and inflammation. Calpain I has been implicated in excitatory amino-acid induced neurotoxicity disorders including ischemia, hypoglycemia, Huntington's Disease, and epilepsy.

The lysosomal cysteine protease cathepsin B has been implicated in the following disorders: arthritis, inflammation, myocardial infarction, tumor metastasis, and muscular dystrophy. Other lysosomal cysteine proteases include cathepsins C, H, L and S. Interleukin-1β converting enzyme ("ICE") is a cysteine protease which catalyzes the formation of interleukin-1β. Interleukin-1β is an immunoregulatory protein implicated in the following disorders: inflammation, diabetes, septic shock, rheumatoid arthritis, and Alzheimer's disease. ICE has also been linked to apoptotic cell death of neurons, which is implicated in a variety of neurodegenerative disorders including Parkinson's disease, ischemia, and amyotrophic lateral sclerosis (ALS).

Cysteine proteases are also produced by various pathogens. The cysteine protease clostripain is produced byClostridium histolyticum. Other proteases are produced byTrypanosoma cruzi, malaria parasites Plasmodiumfalciparum andP.vinckei andStreptococcus. Hepatitis A viral protease HAV C3 is a cysteine protease essential for processing of picornavirus structural proteins and enzymes.

Exemplary serine proteases implicated in degenerative disorders include thrombin, human leukocyte elastase, pancreatic elastase, chymase and cathepsin G. Specifically, thrombin is produced in the blood coagulation cascade, cleaves fibrinogen to form fibrin and activates Factor VIII; thrombin is implicated in thrombophlebitis, thrombosis and asthma. Human leukocyte elastase is implicated in tissue degenerative disorders such as rheumatoid arthritis, osteoarthritis, atherosclerosis, bronchitis, cystic fibrosis, and emphysema. Pancreatic elastase is implicated in pancreatitis. Chymase, an enzyme important in angiotensin synthesis, is implicated in hypertension, myocardial infarction, and coronary heart disease. Cathepsin G is implicated in abnormal connective tissue degradation, particularly in the lung.

Given the link between cysteine and serine proteases and various debilitating disorders, compounds which inhibit these proteases would be useful and would provide an advance in both research and clinical medicine.

Examples of protease inhibitors are disclosed in WO 92/12140 and in WO 98/41506.

SUMMARY OF THE INVENTION

The present invention is directed to selected quinoline-containing α-ketoamide inhibitors of cysteine and serine proteases represented by the general Formula I:wherein:

• X is CH, N, or CQ1, with the proviso that when X is CQ1, at least one of Q or Q1 is H;
• R is selected from the group consisting of H, alkyl having from one to 6 carbons, arylalkyl having from 7 to 15 carbons, cyclo(heteroalkyl) in which the ring contains from 5 to 14 ring atoms, heteroaryl C1-6alkyl in which the heteroaryl ring contains from 5 to 14 ring atoms, C1-10alkoxyC1-10alkyl, a side chain of a naturally occurring amino acid in the R or S configuration, and (CH2)nNH-L, said alkyl, cycloalkyl, arylalkyl, heteroalkyl, and heteroarylalkyl groups being optionally substituted with one or more J groups;
• L is selected from the group consisting of alkoxycarbonyl having from 2 to 7 carbons, arylalkoxycarbonyl in which the arylalkoxy group contains 7 to 15 carbons, S(=O)2R2, and N-nitroimino;
• R2 is selected from the group consisting of C1-6alkyl, and aryl having from 6 to 14 carbons;
• R1 is selected from the group consisting of H, halogen, cyano, nitro, sulfonic acid, hydroxyl, C1-10alkyl, C1-10alkoxy, hydroxymethyl, C1-10alkoxymethyl, arylalkyl having from 7 to 15 carbons, carboxyl, C1-10alkoxycarbonyl. C1-10alkylcarbonyloxy, haloC1-10alkyl, N(RR3), and C1-10alkylcarbonyl;
• R3 is the same as R;
• Q is selected from the group consisting of H, C1-6alkyl, C3-10cycloalkyl, hydroxyl, C1-1alkoxy, halogen, arylalkyl having from 7 to 15 carbons, arylalkenyl having from 8 to 16 carbons, arylalkynyl having from 8 to 16 carbons, aryl having from 6 to 14 carbons, heteroaryl having from 5 to 14 ring atoms, cyclo(heteroalkyl) having from 5 to 14 ring atoms, cycloalkyl having from 3 to 10 carbons, S-R, S(=O)R, S(=O)2R, N(RR3), and NHS(=O)2R, said arylalkyl, arylalkenyl, arylalkynyl, aryl, heteroaryl, and heteroallsyl groups being optionally substituted with one or more J groups;
• Q1 is the same as Q;
• Z is COCONH-R7;
• R7 is selected from the group consisting of K, -A-N(R8)-G, -O-A-N(R8)-G, -A-SO2N(R8)(R9), and -O-A-SO2N(R8)(R9);
• K is selected from the group consisting of C1-10alkyl, alkenyl selected from ethenyl and propenyl, alkynyl selected from ethynyl and propynyl, C3-10cycloalkyl, cyclo(heteroalkyl) having from 5 to 14 ring atoms, heteroaryl having from 5 to 14 ring atoms, C6-14aryl, C7-15arylalkyl, heterocycloalkyl having a ring having from to 14 ring atoms attached through a C1-6alkyl, C1-10alkoxy, C1-10alkoxyC1-10alkyl, C7-15arylalkyloxy, and N(RR3), said K groups being optionally substituted with one or more J groups;
• A is C1-6alkylene optionally substituted with one or more J groups;
• R8 is selected from the group consisting of H and C1-6alkyl;
• R9 is selected from the group consisting of H, C1-10alkyl, C6-14aryl, and heterocyclyl having from 5 to 14 ring atoms, said alkyl, aryl, and heterocyclyl groups being optionally substituted with one or more J groups;
• G is selected from the group consisting of C(=O)C6-14aryl, C(=O)heteroaryl wherein the heteroaryl has from 5 to 14 ring atoms, C(=O)cyclo(heteroall:yl) wherein the cyclo(heteroalkyl) has from 5 to 14 ring atoms, C1-10alkanoyl, C(=S)NH(C6-14aryl), C(=O)NH(C6-14aryl), C(=O)NH(C3-10cycloalkyl), CO2-(C6-14aryl), C(=O)C1-10alkyl, CO2(C1-10alkyl), CO2(C7-15arylalkyl), C1-10alkylsulfonyl, C1-10alkenylsulfonyl, C6-14arylsulfonyl, heteroarylsulfonyl wherein said heteroaryl has from 5 to 14 ring atoms, a side chain of a naturally occurring amino acid in the R or S configuration, a protecting group selected from the group consisting of benzyloxycarbonyl, toluenesulfonyl, t-butoxycarbonyl, methyl ester and benzyl ether groups, and SO2N(RR3), said G groups being optionally substituted with one or more J groups;
• J is selected from the group consisting of H, halogen, cyano, nitro, hydroxyl, C1-10alkyl, C1-10alkoxy, C6-14aryl, C7-15arylakyl C1-10alkoxycarbonyl, C1-10alkylcarbonyloxy, C1-10alkenylcarbonyloxy, haloC1-10alkyl, aminoC1-10alkyl, haloC1-10alkoxy, SO2N(RR3), SO2NH(C6-14aryl), SO2NH(heteroaryl) wherein said heteroaryl has from 5 to 14 ring atoms, NHC(=O)NH(C6-14aryl), NH(C=O)NH(heteroaryl) wherein said heteroaryl has from 5 to 14 ring atoms, NHSO2(C6-14aryl), NHC(=O)C1-10alkyl, NHC(=O)C6-14aryl, NHC(=O)heteroaryl wherein said heteroaryl has from 5 to 14 ring atoms, N(RR3), and NH=C(NH2)2, where R and R3 are not further substituted with J;
• n is an integer from 2 to 6;
• wherein alkyl includes straight-chain, branched and cyclic hydrocarbon groups; or a pharmaceutically acceptable salt thereof;
• with the proviso that when R7 is K, then Q is selected from the group consisting of optionally substituted arylalkenyl and optionally substituted arylalkynyl; and
• with the further proviso that when K is alkyl, then X is not CH when Q is optionally substituted arylalkynyl.

In some preferred embodiments of the compounds of Formula I, X is CH. In further preferred embodiments of the compounds of Formula I, R is selected from the group consisting of alkyl having from 2 to 4 carbons, and arylalkyl, with benzyl being particularly preferred.

In some preferred embodiments of the compounds of Formula I, R¹ is H or alkoxy, with H being preferred.

In some preferred embodiments of the compounds of Formula I, Q is selected from the group consisting of arylalkynyl, aryl and halo, with phenylalkynyl being preferred.

In further preferred embodiments of the compounds of Formula I, A is selected from the group consisting of (CH₂)_n wherein n is 2 or 3, and (CH₂)_vCH₂-J where v is an integer from 1 to 6. When A is (CH₂)_vCH₂-J, v is preferably 2 or 3.

In some preferred embodiments of the compounds of Formula I, K is selected from the group consisting of alkyl, hydroxyalkyl, haloalkyl, alkynyl, heterocycloalkyl, arylalkyl, and heteroalkyl.

In further preferred embodiments of the compounds of Formula I, G is selected from the group consisting of substituted or unsubstituted C(=O)aryl, C(=O)heteroaryl, arylsulfonyl, and heteroarylsulfonyl. Preferably, G is selected from the group consisting of unsubstituted arylsulfonyl, substituted arylsulfonyl, unsubstituted heteroarylsulfonyl and substituted heteroarylsulfonyl.

In some preferred embodiments of the compounds of Formula I, X is CH, Z is COCONH-K, R₁ is H, and R is selected from the group consisting of alkyl having from 2 to 4 carbons and arylalkyl, with benzyl being preferred.

In some especially preferred embodiments of the compounds of Formula I, X is CH, Z is COCONH-K, R₁ is H, R is selected from the group consisting of alkyl having from 2 to 4 carbons and arylalkyl, with benzyl being preferred, and Q is arylalkynyl, with phenylethynyl being preferred.

In some especially preferred embodiments of the compounds of Formula I, Q, X, R¹, R, and Z are selected from the group of substituents listed in Tables 2 to 5,infra. Particularly preferred embodiments of the compounds of Formula I are listed in Tables 2 to 5,infra.

Because the quinoline-containing α-ketoamides of the invention inhibit cysteine proteases and serine proteases, they can be used in both research and therapeutic settings.

In a research environment, preferred compounds' having defined attributes can be used to screen for natural and synthetic compounds which evidence similar characteristics in inhibiting protease activity. The compounds can also be used in the refinement ofin vitro andin vivo models for determining the effects of inhibition of particular proteases on particular cell types or biological conditions.

In a therapeutic setting, given the connection between cysteine proteases and certain defined disorders, and serine proteases and certain defined disorders, compounds of the invention can be utilized to alleviate, mediate, reduce and/or prevent disorders which are associated with abnormal and/or aberrant activity of cysteine proteases and/or serine proteases.

In preferred embodiments, compositions are provided for inhibiting a serine protease or a cysteine protease comprising a compound of the invention. In other preferred embodiments, methods are provided for inhibiting serine proteases or cysteine proteases comprising contacting a protease selected from the group consisting of serine proteases and cysteine proteases with an inhibitory amount of a compound of the invention.

Methodologies for making the present quinoline-containing α-ketoamide inhibitors are also disclosed. These and other features of the compounds of the subject invention are set forth in more detail below.

DETAILED DESCRIPTION

Disclosed herein are selected quinoline-containing α-ketoamide serine and cysteine protease inhibitors, which are represented by the following Formula I:wherein:

• X is CH, N, or CQ1, with the proviso that when X is CQ1, at least one of Q or Q1 is H;
• R is selected from the group consisting of H, alkyl having from one to about 6 carbons, arylalkyl having from about 7 to about 15 carbons, heteroalkyl in which the ring contains from about 5 to about 14 ring atoms, heteroarylalkyl in which the heteroaryl ring contains from about 5 to about 14 ring atoms, alkoxyalkyl, a side chain of a naturally occurring amino acid in the R or S configuration, and (CH2)nNH-L, said alkyl, arylalkyl, heteroalkyl, and heteroarylalkyl groups being optionally substituted with one or more J groups;
• L is selected from the group consisting of alkoxycarbonyl having from 2 to about 7 carbons, arylalkoxycarbonyl in which the arylalkoxy group contains about 7 to about 15 carbons, S(=O)2R2, and N-nitroimino;
• R2 is selected from the group consisting of lower alkyl, and aryl having from about 6 to about 14 carbons;
• R1 is selected from the group consisting of H, halogen, cyano, nitro, sulfonic acid, hydroxyl, alkyl, alkoxy, hydroxymethyl, alkoxymethyl, arylalkyl, carboxyl, alkoxycarbonyl, alkylcarbonyloxy, haloalkyl, N(RR3), and acyl;
• R3 is the same as R;
• Q is selected from the group consisting of H, lower alkyl, cycloalkyl, hydroxyl, alkoxy, halogen, arylalkyl having from about 7 to about 15 carbons, arylalkenyl having from about 8 to about 16 carbons, arylalkynyl having from about 8 to about 16 carbons, aryl having from about 6 to about 14 carbons, heteroaryl having from about 5 to about 14 ring atoms, heteroalkyl having from about 5 to about 14 ring atoms, cycloalkyl having from about 3 to about 10 carbons, S-R, S(=O)R, S(=O)2R, N(RR3), and NHS(=O)2R, said arylalkyl, arylalkenyl, arylalkynyl, aryl, heteroaryl, and heteroalkyl groups being optionally substituted with one or more J groups;
• Q1 is the same as Q;
• Z is COCONH-R7;
• R7 is selected from the group consisting of K, -A-N (R8)-G, -O-A-N (R8) -G, A-SO2N (R8) (R9), and -O-A-SO2N (R8) (R9) ;
• K is selected from the group consisting of alkyl, alkenyl, alkynyl, cycloalkyl, heteroalkyl, heteroaryl, aryl, arylalkyl, heterocycloalkyl, alkoxy, alkoxyalkyl, arylalkyloxy, and N(RR3), said K groups being optionally substituted with one or more J groups;
• A is lower alkylene optionally substituted with one or more J groups;
• R8 is selected from the group consisting of H and lower alkyl;
• R9 is selected from the group consisting of H, alkyl, aryl, and heterocyclyl, said alkyl, aryl, and heterocyclyl groups being optionally substituted with one or more J groups;
• G is selected from the group consisting of C(=O) aryl, C(=O) heteroaryl, C(=O) heteroalkyl, alkanoyl, C(=S) NH(aryl), C (=O) NH (aryl), C (=O) NH (cycloalkyl), CO2 (aryl), C(=O) alkyl, CO2 (alkyl), CO2 (arylalkyl), alkylsulfonyl, alkenylsulfonyl, arylsulfonyl, heteroarylsulfonyl, a side chain of a naturally occurring amino acid in the R or S configuration, a blocking group, and SO2N(RR3), said G groups being optionally substituted with one or more J groups;
• J is selected from the group consisting of H, halogen, cyano, nitro, hydroxyl, alkyl, alkoxy, aryl, arylalkyl, alkoxycarbonyl, alkylcarbonyloxy, alkenylcarbonyloxy, haloalkyl, aminoalkyl, haloalkoxy, SO2N (RR3), SO2NH (aryl) SO2NH (heteroaryl), NHC (=O) NH (aryl), NH (C=O) NH (heteroaryl), NHSO2 (aryl), NHC (=O) alkyl, NHC (=O) aryl, NHC (=O) heteroaryl, N(RR3) where R and R3 are not further substituted with J, and NH=C(NH2)2;
• n is an integer from 2 to 6; or a pharmaceutically acceptable salt thereof;
• with the proviso that when R7 is K, then Q is selected from the group consisting of optionally substituted arylalkenyl and optionally substituted arylalkynyl; and
• with the further proviso that when K is alkyl, then X is not CH when Q is optionally substituted arylalkynyl.

It is recognized that various stereoisomeric forms of the compounds of Formula I may exist. Preferred compounds of the invention have the L-configuration at the carbon to which the substituent R is attached. However, reacemates and individual enantiomers and mixtures thereof form part of the present invention.

As used herein, the term "quinoline" denotes a "quinoline" or a "quinoline N-oxide" structure.

In the compounds of Formula I, where a bond to a substituent is shown to cross the bond connecting two atoms in a ring, it is intended that such substituent may be bound to any atom in the ring.

As used herein, the term "alkyl" includes straight-chain, branched and cyclic hydrocarbon groups such as, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, 1-ethylpentyl, hexyl, octyl, cyclopropyl, methylcyclopentyl, cyclohexyl, and adamantane groups. Preferred alkyl groups have 1 to about 10 carbon atoms, with 1 to about 6 carbon atoms (i.e., "lower alkyl") being preferred. "Lower alkylene" groups are branched or unbranched alkylene groups of 1 to about 6 carbon atoms such as, for example, ethylene (-CH₂CH₂-), propylene, butylene, hexylene, 1-methylethylene, 2-methylethylene, and 2-methylpropylene. "Acyl" (i.e., "alkanoyl") groups are alkylcarbonyl groups. "Aryl" groups are aromatic cyclic compounds including but not limited to phenyl, tolyl, naphthyl, anthracyl, phenanthryl, pyrenyl, biphenyl, and xylyl. Preferred aryl groups include phenyl and naphthyl. The term "carbocyclic", as used herein, refers to cyclic groups in which the ring portion is composed solely of carbon atoms. The term "halogen" refers to F, Cl, Br, and I atoms. The term "arylalkyl" (or "aralkyl") denotes alkyl groups which bear aryl groups, for example, benzyl groups.

As used herein, "alkoxy" groups are alkyl groups linked through an oxygen atom. Examples of alkoxy groups include methoxy (-OCH₃) and ethoxy (-OCH₂CH₃) groups. In general, the term "oxy" when used as a suffix denotes attachment through an oxygen atom. Thus, alkoxycarbonyl groups are carbonyl groups which contain an alkoxy substituent, i.e., groups of general formula -C(=O)-O-R, where R is alkyl. The term "alkoxyalkyl" denotes an alkoxy group attached to an aklyl group. The term "aryloxy" denotes an aryl group linked through an oxygen atom, and the term "arylalkyloxy" denotes an arylalkyl group linked through an oxygen atom.

As used herein, the term "alkenyl" is intended to include straight-chain or branched hydrocarbon chains having at least one carbon-carbon double bond. Examples of alkenyl groups include ethenyl and propenyl groups. Arylalkenyl groups are alkenyl groups that have one or more aryl groups appended thereto. As used herein, the term "alkynyl" is intended to include straight-chain or branched hydrocarbon chains having at least one carbon-carbon triple bond. Examples of alkynyl groups include ethynyl and propynyl groups. Arylalkynyl groups are alkynyl groups that have one or more aryl groups appended thereto.

The terms "heterocycle", "heterocyclyl", and "heterocyclic" refer to cyclic groups in which a ring portion includes at least one heteroatom such as O, N or S. Heterocyclic groups include "heteroaryl" as well as "heteroalkyl" groups. Preferred "heteroaryl" groups include pyridyl, pyrimidyl, pyrrolyl, furyl, thienyl, imidazolyl, triazolyl, tetrazolyl, quinolyl, isoquinolyl, benzimidazolyl, thiazolyl, bipyridyl, phthalimido, and benzothiazolyl. The term "heterocycloalkyl" denotes a heterocycle attached through a lower alkyl group. The term "heteroaryl" denotes aryl groups having one or more hetero atoms contained within an aromatic ring. The term "heteroarylalkyl" denotes a heteroaryl group attached through an alkyl group. The term "heteroalkyl" denotes a heterocyclic group which contains at least one saturated carbon atom in the heterocyclic ring. Examples of heteroalkyl groups include piperidine, dihydropyridine, tetrahydroisoquinyl, and ε-caprolactam groups.

As used herein, the term "amino acid" denotes a molecule containing both an amino group and a carboxyl group. As used herein the term "L-amino acid" denotes an α-amino acid having the L- configuration around the α-carbon, that is, a carboxylic acid of general formula CH (COOH) (NH₂) - (side chain), having the L-configuration. The term "D-amino acid" similarly denotes a carboxylic acid of general formula CH(COOH) (NH₂)-(side chain), having the D-configuration around the α-carbon. Side chains of L-amino acids include naturally occurring and non-naturally occurring moieties. Nonnaturally occurring (i.e., unnatural) amino acid side chains are moieties that are used in place of naturally occurring amino acid sidechains in, for example, amino acid analogs. See, for example, Lehninger,Biochemistry, Second Edition, Worth Publishers, Inc, 1975, pages 73-75. One representative amino acid side chain is the lysyl side chain, -(CH₂)₄-NH₂. Other representative α-amino acid side chains are shown below in Table 1.

The disclosed compounds of the invention are useful for the inhibition of cysteine proteases and serine proteases. As used herein, the terms "inhibit" and "inhibition" mean having an adverse effect on enzymatic activity. An inhibitory amount is an amount of a compound of the invention effective to inhibit a cysteine and/or serine protease.

Pharmaceutically acceptable salts of the cysteine and serine protease inhibitors also fall within the scope of the compounds as disclosed herein. The term "pharmaceutically acceptable salts" as used herein means an inorganic acid addition salt such as hydrochloride, sulfate, and phosphate, or an organic acid addition salt such as acetate, maleate, fumarate, tartrate, and citrate. Examples of pharmaceutically acceptable metal salts are alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as magnesium salt and calcium salt, aluminum salt, and zinc salt. Examples of pharmaceutically acceptable organic amine addition salts are salts with morpholine and piperidine. Examples of pharmaceutically acceptable amino acid addition salts are salts with lysine, glycine, and phenylalanine.

Compounds provided herein can be formulated into pharmaceutical compositions by admixture with pharmaceutically acceptable excipients and carriers. As noted above, such compositions may be prepared for use in parenteral administration, particularly in the form of liquid solutions or suspensions; or oral administration, particularly in the form of tablets or capsules; or intranasally, particularly in the form of powders, nasal drops, or aerosols; or dermally, via, for example, transdermal patches; or prepared in other suitable fashions for these and other forms of administration as will be apparent to those skilled in the art.

The composition may conveniently be administered in unit dosage form and may be prepared by any of the methods well known in the pharmaceutical art, for example, as described in Remington'sPharmaceutical Sciences (Mack Pub. Co., Easton, PA, 1980). Formulations for parenteral administration may contain as common excipients sterile water or saline, polyalkylene glycols such as polyethylene glycol, oils and vegetable origin, hydrogenated naphthalenes and the like. In particular, biocompatible, biodegradable lactide polymer, lactide/glycolide copolymer, or polyoxyethylene-polyoxypropylene copolymers may be useful excipients to control the release of the active compounds. Other potentially useful parenteral delivery systems for these active compounds include ethylene-vinyl acetate copolymer particles, osmotic pumps, implantable infusion systems, cyclodextrins and liposomes. Formulations for inhalation administration contain as excipients, for example, lactose, or may be aqueous solutions containing, for example, polyoxyethylene-9-lauryl ether, glycocholate and deoxycholate, or oily solutions for administration in the form of nasal drops, or as a gel to be applied intranasally. Formulations for parenteral administration may also include glycocholate for buccal administration, a salicylate for rectal administration, or citric acid for vaginal administration. Formulations for transdermal patches are preferably lipophilic emulsions.

The materials for this invention can be employed as the sole active agent in a pharmaceutical or can be used in combination with other active ingredients which could facilitate inhibition of cysteine and serine proteases in diseases or disorders.

The concentrations of the compounds described herein in a therapeutic composition will vary depending upon a number of factors, including the dosage of the drug to be administered, the chemical characteristics (e.g., hydrophobicity) of the compounds employed, and the route of administration. In general terms, the compounds of this invention may be provided in effective inhibitory amounts in an aqueous physiological buffer solution containing about 0.1 to 10% w/v compound for parenteral administration. Typical dose ranges are from about 1µg/kg to about 1 g/kg of body weight per day; a preferred dose range is from about 0.01 mg/kg to 100 mg/kg of body weight per day. Such formulations typically provide inhibitory amounts of the compound of the invention. The preferred dosage of drug to be administered is likely, however, to depend on such variables as the type or extent of progression of the disease or disorder, the overall health status of the particular patient, the relative biological efficacy of the compound selected, and formulation of the compound excipient, and its route of administration.

As used herein, the term "contacting" means directly or indirectly causing at least two moieties to come into physical association with each other. Contacting thus includes physical acts such as placing the moieties together in a container, or administering moieties to a patient. Thus, for example administering a compound of the invention to a human patient evidencing a disease or disorder associated with abnormal and/or aberrant activity of such proteases falls within the scope of the definition of the term "contacting".

The invention is further illustrated by way of the following examples which are intended to elucidate the invention. These examples are not intended, nor are they to be construed, as limiting the scope of the disclosure.

ExamplesExamples 1-158:

The compounds shown in Tables 2 through 5 were prepared using conditions described in the "General Methods" section below. Enzyme inhibitory activity (IC₅₀) was determined as described in Examples 159 and 160.

In all Examples, R¹ is H unless otherwise noted.

General Methods:

Thin layer chromatography was performed using silica gel coated plates (MK6F 60A, size 1 x 3 in, layer thickness 250 µm, Whatman Inc.). Preparative thin layer chromatography was performed using silica gel coated plates (size 20 x 20.in, layer thickness 1000 micron, Analtech). Preparative column chromatography was carried out using Merck silica gel, 40-63µm, 230-400 mesh.¹H NMR spectra were recorded on a GE QE300 Plus spectrometer at 300 MHZ using tetramethylsilane as internal standard. Electrospray mass spectra were recorded on a VG platform II instrument (Fisons Instruments).

Compounds were prepared following one of the general methods A, B, C or D.

Preparation of Compound 2

A mixture of compound of formula 1 (20 g, 0.106 mol) and phosphorous oxychloride (50 mL, 0.54 mol) was held at reflux for 3 hours. After cooling, the reaction mixture was slowly poured into ice-water and a white precipitate was formed. The precipitate was filtered, thoroughly washed with cold water, and redissolved in ethyl acetate (300 mL). The solution was dried over anhydrous sodium sulfate. Filtration and solvent removal gave 14.78 g of compound2 which was used without further purification.Compound2: tan solid;¹H-NMR (DMSO-d₆) δ 8.60 (d, 1H), 8.00 (d, 1H), 7.90 (s, 1H), 7.80 (t, 1H), 7.70 (t, 1H). MS m/e 208 and 210(M+H with isotopes of chlorine).

Preparation of Compound 3

A mixture of compound2 (20 g, 0.0966 mol), phenylacetylene (13.02 g, 14 mL, 0.127 mol), (PPh₃) PdCl₂ (1.40 g, 0.00193 mol), CuI (0.42 g, 0.00386 mol), triethylamine (19.66 g, 27 mL, 0.192 mol) and anhydrous DMSO (150 mL) was heated at 60-70 °C for 3 hours. After cooling, the reaction mixture was slowly poured into ice-water (300 mL). The aqueous solution was acidified with 2 N HCl and extracted into methylene chloride (3 x 700 mL). The organic layer was washed with brine (1 x 250 mL), dried (anhydrous sodium sulfate) and concentrated to give a residue which, on recrystallization from methylene chloride-hexanes, yielded 23.8 g of compound3.Compound3: white solid;¹H-NMR (DMSO-d₆) δ 8.60 (d, 1H), 8.10 (d, 1H), 8.05 (s, 1H), 7.85 (t, 1H), 7.70 (m, 3H), 7.45 (m, 3H) . MS m/e 274(M+H).

Preparation of Compound 4a

Compound 4a, and related hydroxy-acids used in this study, were synthesized following a general procedure of Harbeson et al.,J. Med. Chem.1994,37, 2918-2929.

Preparation of Compound 4b

To a cooled (-10 °C) solution of compound4a (4.30 g, 0.015 mol) in anhydrous methanol (50 mL) was added slowly thionyl chloride (3.20 mL). After 0.5 hour, the cooling bath was removed, the mixture was stirred for an additional 16 hours and concentrated to give a residue. Trituration with ethyl acetate (30 mL) gave a white solid. The solid was separated by filtration and dried to give 3.50 g of compound4b which was used directly in the next step; MS m/e 210(M+H).

Preparation of Compound 5 (R=Benzyl)

To a cooled (0 °C) solution of compound3 (0.88 g, 0.0032 mol) in anhydrous DMF (10 mL) was added N-methylmorpholine (0.98 g, 0.0096 mol) followed by 1-HOBt (0.43 g, 0.0032 mol) and BOP (1.70 g, 0.0039 mol). The mixture was stirred for 15 minutes and to it was added compound4b (0.95 g, 0.0039 mol). The cooling bath was removed and the mixture was stirred for 4 hours, poured into ice-water (40 mL) and extracted into ethyl acetate (3 x 40 mL). The organic layer was washed with 2% citric acid solution (2 x 40 mL), 2% sodium bicarbonate solution (2 x 40 mL), brine (1 x 50 mL) and dried over anhydrous sodium sulfate. Solvent evaporation under reduced pressure gave a crude material which was purified by flash column chromatography (eluant: 40% ethyl acetate in hexanes) to produce 1.10 g of compound5.Compound5 (Diastereomeric mixture): white solid;¹H-NMR (CDCl₃) δ 8.10 (d, 1H), 7.80-7.20 (2 sets of m, 14H), 6.40 (2 sets of d, 1H), 5.00 (m, 1H), 4.60 and 4.30 (2 sets of t, 1H), 3.85 and 3.75 (2 singlets, 3H), 3.50 and 3.35 (2 sets of d, 1H), 3.10 and 3.00 (2 sets of dd, 2H). MS m/e 465 (M+H) .

Preparation of Compound 6 (R=Benzyl)

To a cooled (0 °C) solution of compound5 (4.33 g, 9.33 mmol) in 1:1 anhydrous methylene chloride and anhydrous acetonitrile (60 mL) was slowly added Dess-Martin periodinane reagent (7.90 g, 18.66 mmol). The cooling bath was removed and the mixture was stirred for an additional 2 hours. The mixture was then diluted with methylene chloride (50 mL) and washed with 10% sodium thiosulfate solution (5 x 50 mL), saturated sodium bicarbonate solution (2 x 50 mL), and brine (1 x 50 mL). Drying (anhydrous sodium sulfate) and solvent removal under reduced pressure gave a residue which was purified by flash column chromatography (eluant: 1:1 EtOAc-hexanes) to yield 3.3 g of compound6.Compound6: white solid;¹H-NMR (CDCl₃) δ 8.20-7.20 (m, 15H), 6.65 (d, 1H), 5.80 (q, 1H), 3.95 (s, 3H), 3.50 (dd, 1H), 3.20 (dd, 1H). MS m/e 463(M+H).

Preparation of Compound 7 (R=Benzyl)

A mixture of compound6, (1.20 g, 2.60 mmol), 1 N NaOH (6.5 mL) and MeOH (15 mL) was stirred at room temperature for 1.5 hours. TLC (50% EtOAc in methylene chloride) showed the complete disappearence of compound6. The reaction mixture was concentrated at the rotavapor and redissolved in water (25 mL). The aqueous layer was washed with ether (2 x 15 mL) and acidified with 1 N HCl. The aqueous layer was then extracted into EtOAc (3 x 50 mL) and the combined ethyl acetate layer was washed with brine (1 x 20 mL), dried (MgS04) and concentrated at the rotavapor to give 1.17 g of compound7.¹H-NMR (CDCl₃) of an aliquot showed absence of a COOCH₃ peak at δ 3.95; MS m/e 449 (M+H) .

Preparation of Compound 8

Compound8 was prepared, for example, by coupling compound7 with an amine in the presence of NMM/HOBt/BOP/DMF, as described for the synthesis of compound5. Purification was achieved by passing a solution of the crude material in methylene chloride or ethyl acetate through Sep-Pak® Vac 6cc (1 g) silica cartidge (Waters Corporation, Milford, MA) and eluting with methylene chloride followed by mixtures of methylene chloride and ethyl acetate.

Harbeson et al. reported (J. Med. Chem.1994,37, 2918-2929) that silica gel chromatography (of a ketoamide) epimerizes the chiral center at P₁.

In General Method B, compounds of the formula5 from General Method A were hydrolyzed to compounds of formula9, following the same procedure as described for the synthesis of compound7. Compounds of formula9 were then coupled with an amine (NMM/HOBt/BOP/DMF), as described for the synthesis of compound5, to produce the α-hydroxyamides (compound10). Dess-Martin oxidation of compounds of formula10 generated the ketoamides (compound8) of the invention, which were used as such or recrystallized from organic solvents.

In General Method C, compound4a (General Method A) was initially coupled to an amine (NMM/HOBt/BOP/DMF), as described for the synthesis of compound 5, to generate compound11. tBoc-deprotection was carried out under standard conditions (4 N HCl in dioxane, room temperature) to generate the amine salt, compound12. Coupling of compound12 with compound3 (NMM/HOBt/BOP/DMF) as described for the synthesis of compound5 produced compound10.

Dess-Martin oxidation of compound10 generated compound8.

In General Method D, compound11 from General Method C was oxidized to generate compound13, which on tBoc-deprotection generated compound14. Coupling of compound3 with compound14 produced the ketoamide of the invention (compound8).

It should be noted that although the General Methods A, B, C and D display only 2-phenylethynylquinoline(3) as the quinoline component of the invention, they are also valid for all other quinoline-4-carboxylic acids shown. Similarly, the methods are also valid for the different hydroxyacids derived from Leu, Nle(4c), Nva(4e), or by extension to any α-hydroxy-β-amino acid, in place of compound4a (derived from phenylalanine).

Preparation of IntermediatesPreparation of Compounds 15 and 16.

Compound3 (1.00 g, 3.66 mmol) was dissolved in DMF (14 mL), and the solution was stirred and hydrogenated at atmospheric pressure over 10% Pd-C (170 mg) for 50 hours (more 10% Pd-C (230 mg total) was added after 23 and 29 h). After filtration, the solvent was evaporated in vacuo at 40 °C. The residue was dissolved in methylene chloride, and the solution was rinsed twice with water and twice with brine, then dried over anhydrous MgSO₄. Evaporation of the solvent afforded a crude mixture of compounds15 and16 (700 mg) as a brownish-orange semisolid. Compound15 was purified by recrystallization from methanol. Compound16 was purified from the resulting mother liquor by preparative TLC (eluent: CH₂Cl₂ - MeOH - HOAc, 94:5:1).Compound15: MS m/e 276 (M+H).Compound16:¹H NMR (DMSO-d₆) δ 8.64 (d, 1H), 8.08 (d, 1H), 7.92 (s, 1H), 7.82 (t, 1H), 7.70 (t, 1H), 7.38 (m, 5H), 3.30 (t, 2H), 3.16 (t, 2H); MS m/e 278 (M+H).

The synthesis of a representative example of an amine containing a terminal sulfonamide moiety is shown in General Method E.

Preparation of Compound 18

To a solution of 1,2-ethylenediamine (compound17, 10.80 g, 12.00 mL, 0.18 mol) in THF (30 mL) was added slowly BOC-ON (22.10 g, 0.09 mol) in THF (70 mL) over a period of 4 hours. The reaction mixture was stirred overnight, concentrated on a rotavapor, and the residue was dissolved in water (150 mL). The aqueous layer was acidified (pH ~ 5-6) with solid citric acid monohydrate, washed with ether (3 x 50 mL), and then treated (at 0 °C) with 6 N NaOH solution to pH ∼ 12-13. The basic solution was extracted into ethyl acetate (3 x 100 mL), and the combined ethyl acetate layer was dried (MgSO₄) and concentrated to generate 7.23 g of monoprotected diamine, compound18.Compound18: semisolid;¹H-NMR (CDCl₃) δ 5.00 (broad, 1H), 3.20 (broad q, 2H), 2.80 (t, 2H), 1.45 (s, 9H), 1.25 (broad, 2H) .

Preparation of Compound 19

A cooled (0-5 °C) solution of the compound18 (0.321 g, 0.002 mol) in methylene chloride (5 mL) was treated sequentially with triethylamine (0.243 g, 0.33 mL, 0.0024 mol) and benzenesulfonyl chloride (0.423 g, 0.30 mL, 0.0024 mol). The ice-bath was removed and the mixture was stirred for an additional 0.5 hour, washed successively with water (2 x 5 mL), cold (0-5 °C) 0.5 N HCl (1 x 5 mL), 2% NaHCO₃ solution (1 x 5 mL), and brine (1 x 5 mL). The solution was dried (MgSO₄), and the solvent was evaporated to give a residue which was washed several times with n-pentane to give 0.60 g of the sulfonamide derivative, compound19.Compound19: white solid, mp 92-95 °C; R_f (TLC, 5% methanol in methylene chloride) 0.55;¹H-NMR (CDCl₃) δ 7.85 (d, 2H), 7.55 (m, 3H), 5.30 (broad d, 1H), 4.85 (broad, 1H), 3.25 (broad q, 2H), 3.10 (broad q, 2H), 1.40 (s, 9H).

Preparation of Compound 20

A solution of compound19 (0.560 g, 0.0019 mol) in 1,4-dioxane (4 mL) was treated with 4 N HC1 in dioxane (4 mL). The mixture was stirred at room temperature for 1 hour and concentrated at the rotavapor. The residue was washed several times with ethyl acetate and dried under vacuum to give 0.40 g of the amine salt, compound20.Compound20: white solid, mp 178-180 °C;¹H-NMR (DMSO-d₆) δ 8.20-8.00 (broad t, 4H), 7.80 (d, 2H), 7.60 (m, 3H), 2.95 (broad q,_ 2H), 2. 80 (broad, 2H).

The synthesis of a representative alkoxyamine, containing a terminal sulfonamide moiety, is shown in General Method F.

Preparation of Compound 23

To a solution of benzohydroxamic acid (compound21, 5.00 g, 0.0365 mol) in DMF (50 mL) was slowly added sodium methoxide (2.50 g, 0.044 mol). The mixture was stirred for 10 minutes and to it was added N-(2-bromoethyl)phthalimide (compound22, 9.00 g, 0.0333 mol). The reaction mixture was then stirred overnight, concentrated at the rotavapor, and partitioned between methylene chloride (200 mL) and 0.1 N NaOH (200 mL). The organic layer was separated, washed with water (2 x 30 mL), dried (MgSO₄) and concentrated to a small volume. Trituration with ethanol produced 4.30 g of compound23 which was used without further purification; MS m/e 311 (M+H).

Preparation of Compound 24

A mixture of compound23 (1.00 g, 0.0032 mol), hydrazine (1 mL) and 95% ethanol was held at reflux for 6 hours. After cooling, the reaction mixture was filtered and the filtrate was concentrated to give 0.575 g of compound24 which was directly used in the next step; MS m/e 181(M+H).

Preparation of Compound 25

Compound25 was generated from compound24 following the same procedure as described for the synthesis of Compound19 (General Method E); the crude product was purified by flash column chromatography (eluant 20% ethyl acetate in methylene chloride) to give 0.75 g of compound25; MS m/e 321 (M+H) .

Preparation of Compound 26

A mixture of compound25 (0.60 g, 1.87 mmol) and 6 N HC1 (20 mL) was held at reflux for 3 hours, cooled to room temperature and filtered. The filtrate was concentrated in vacuo overnight, to generate the amine salt, compound26; MS m/e 217 (M+H).

Preparation of Compound 27

2-Phenylquinazoline-4-carboxylic acid (compound27) was prepared following a general procedure of Giardina et al,J. Med. Chem.1997,40, 1794-1807. This material can be incorporated into General Method A replacing compound3.

Preparation of the Compound of Example 11 byGeneral Method A:

17: pale yellow solid;¹H-NMR (CDCl₃) δ 8.10 (d, 1H), 7.95 (d, 1H), 7.75 (t, 1H), 7.70-7.10 (a series of m, 13H), 6.55 (d, 1H), 5.90 (m, 1H), 3.65-3.10 (a series of m, 6H), 3.40 (s, 3H). MS m/e 506 (M+H).

Preparation of the Compound of Example 96 byGeneral Method B:

105: pale yellow solid;¹H-NMR (CDCl₃) δ 8.10 (d, 1H), 7.95 (d, 1H), 7.80-7.10 (a series of m, 19H), 6.55 (d, 1H), 5.90 (m, 1H), 5.10 (t, 1H),3.50 (m, 3H), 3.20 (m, 3H). MS m/e 631 (M+H) .

Preparation of the Compound of Example 144 byGeneral Method C

159: pale yellow solid;¹H-NMR (DMSO-d₆) δ 9.35 (d, 1H), 8.90 (t, 1H), 8.50 (d, 1H), 8.10 (t, 1H), 8.00-7.70 (m, 7H), 7.55 (t, 2H), 7.40-7.10 (m, 6H), 5.50 (m, 1H), 3.30 (m, 2H), 3.00 (q, 3H), 2.75 (m, 1H). MS m/e 648 and 650(M+H, with different isotopes of chlorine), 670 and 672(M+Na, with different isotopes of chlorine).

Example 159Inhibition of Cysteine Protease Activity

To evaluate inhibitory activity, stock solutions (40 times concentrated) of each compound to be tested were prepared in 100% anhydrous DMSO and 5 µl of each inhibitor preparation was aliquoted into each of three wells of a 96-well plate. Recombinant human calpain I, prepared by the method of Meyer et al. (Biochem. J.1996, 314: 511-519), was diluted into assay buffer (i.e., 50mM Tris, 50mM NaCl, 1mM EDTA, 1mM EGTA, and 5mM ß-mercaptoethanol, pH 7.5, including 0.2mM Succ-Leu-Tyr-MNA), and 175 µl was aliquoted into the same wells containing the independent inhibitor stocks as well as to positive control wells containing 5 µl DMSO, but no compound. To start the reaction, 20 µl of 50 mM CaCl₂ in assay buffer was added to all wells of the plate, excepting three, which were used as background signal baseline controls. Substrate hydrolysis was monitored every 5 minutes for a total of 30 minutes. Substrate hydrolysis in the absence of inhibitor was linear for up to 15 minutes.

Inhibition of calpain I activity was calculated as the percent decrease in the rate of substrate hydrolysis in the presence of inhibitor relative to the rate in its absence. Comparison between the inhibited and control rates was made within the linear range for substrate hydrolysis. The IC₅₀s of inhibitors (concentration yielding 50% inhibition) were determined from the percent decrease in rates of substrate hydrolysis in the presence of five to seven different concentrations of the test compound. The results were plotted as percent inhibition versus log inhibitor concentration, and the IC₅₀ was calculated by fitting the data to the four-parameter logistic equation shown below using the program GraphPad Prism (GraphPad Software, Inc., San Diego, CA.).$y=d+\left[\left(a-d\right)/\left(1+{\left(x/c\right)}^b\right)\right]$

The parameters a, b, c, and d are defined as follows: a is % inhibition in the absence of inhibitor, b is the slope, c is the IC₅₀, and d is the % inhibition at an infinite concentration of inhibitor.

Results are presented Tables 2-5.

To demonstrate activity against another cysteine protease, cathepsin B (Calbiochem, cat#219364), asays were performed substantially the same as outlined above except that the cathepsin B was diluted into a different assay buffer consisting of 50mM sodium acetate (pH 6.0)/1mM EDTA/1mM dithiothreitol and the substrate used was 0.1mM Cbz-Phe-Arg-AMC (Bachem cat# I-1160). Additionally, the order of reagents added to the plate was altered because the enzyme is consitutively active. Following inhibitor addition to the plates, appropriate 2x concentrated stock dilutions of the enzyme preparations were made in assay buffer and 100µl added to each well. The assay was initiated by addition of 100µl of 2x concentrated stock dilution of substrate in assay buffer. Substrate hydrolysis was monitored using a Fluoriskan II (ex=390 nm; em=460 nm). Results are presented in Table 6.

Example 160Inhibition of Serine Protease Activity

To demonstrate activity against the serine protease α-chymotrypsin (Sigma Chem. Co. Cat. #C-3142) the protocol of Example 159 was followed except that the enzyme was diluted into assay buffer consisting of 50mM Hepes (pH 7.5)/0.5M NaCl and the final substrate concentration used was 0.03mM Succ-Ala-Ala-Pro-Phe-AMC (Bachem, Inc. Cat. #I-1465). Additionally, because α-chymotrypsin is not a calcium sensitive enzyme and is constitutively active, following addition of inhibitor stocks to the 96 well plates, 100µl of a 2-fold concentrated stock of enzyme in dilution buffer was first added and the reaction was started by addition of 100µl of a 2-fold concentrated stock of substrate in assay buffer. Substrate hydrolysis was monitored every 5 minutes up to 30 minutes using a Fluoroskan II (ex=390nm em=460nm). Results are listed in Table 6.

Q = Phenylethynyl

X = CH

				MH+
1		(91%)	A	486
2		(98%)	A	530
3		210	A	488
4		(100%)	A	502
5		(95%)	A	501
6		(89%)	A	556
7		110	A	536
8		570	A	564
9		230	A	664
10		630	A	536
11		480	A	506
12		(94%)	A	506
13		(100%)	A	520
14		(98%)	A	534
15	adamantyl	(41%)	A	582
16	cyclopropane	(100%)	A	488
17	(1-benzyl)piperidin4-yl	638	A	621
18	(2-methylcyclohexyl)	(76%)	A	544
19	(4-methylcyclohexyl)	(91%)	A	544
20	(3-methylcyclohexyl)	(94%)	A	544
21		(100%)	A	603
22	ε-caprolactam	78	A	559
23		(75%)	A	582
24		(21%)	A	582
25		(42%)	A	805
26		(31%)	A	602
27		510	A	552
28	cyclohexan-2-ol	(61%)	A	546
29	[trans]cyclohexan4-ol	296	A	546
30	cyclohexyl	(85%)	A	530
31		(27%)	A	546
32	tetrahydronaphth-1-yl	(31%)	A	578
33	piperon-5-yl	388	A	582
34		227	A	580
35	benzodioxan-6-yl	(71%)	A	582
36		79*	A	538
37		(95%)	A	598
38		(92%)	A	598
39		184	A	617
40		(95%)	A	583
41		(92%)	A	612
42		184	A	631
43		(95%)	A	552
44		(88%)	A	628
45		69	A	540
46		390	A	540
47		(85%)	A	540
48		(95%)	A	601
49		160	A	621
50		(84%)	A	556
51		424	A	575
52		366	A	573
53		226	A	621
54		(84%)	A	532
55	indan-2-yl	(60%)	A	564
56		146	A	613
57		(71%)	A	625
58		98	A	585
59		248	A	604
60		140	A	533
61		(94%)	A	689
62		190	A	640
63		257	A	668
64		67	A	610
65		(100%)	A	595
66		113	A	656
67		93	A	671
68		260	A	676
69		47	A	649
70		137	A	635
71		89	A	689
72		(96%)	A	699
73		220	A	695
74		132	A	656
75		100	A	699
76		(98%)	A	650
77		77	A	656
78		(88%)	A	676
79		43	A	688
80		200	A	694
81		62	A	661
82		70	A	656
83		70	A	646
84		26	A	676
85		48	A	770
86		31	A	712
87		56*	A	632
88		41	A	681
89		130	A	682
90		76	A	637
91		42	A	657
92		94	A	645
93		180	A	569
94		141	A	598
95		43	A	631
96		29*	A	631
97		(100%)	A	605
98		126	A	624
99		130	A	583
100		55	A	645
101		(84%)	A	771
102		82	A	538
103		65	A	492
104	OBn	60	A	554
105		128	A	647
106		(97%)	A	691
107		(93%)	A	576
108		(94%)	A	728
109		655	A	610
110		(43%)	A	491
111		(95%)	A	560

Q = phenylethynyl

X = CH

					MH+
112		iBu	(95%)	B	472
113		iBu	(92%)	A	502
114		iBu	(93%)	A	502
115		iBu	(100%)	B	591
116		iBu	(89%)	A	576
117		iBu	(82%)	A	597
118		iBu	120	B	666
119		iBu	(80%)	A	611
120		Et	(92%)	A	444
121		Et	(94%)	A	474
122		Et	(89%)	A	569
123		Et	58	A	637
124		Et	80	A	583
125		Et	(32%)	A	548
126		Bu	39	A	665
127		Bu	29	A	642
128		Pr	187	A	651
129		Pr	27	A	628

X = CH

					MH+
130	PhCH=CH	Bu	105	D	506
131	H		(97%)*	C	531
132	H(N-oxide)		(96%)	D	547
133	HO		(95%)	D	547
134			(17%)	D	434
135	Piperidin-1-yl		(91%)	D	624
136	Pyridin-3-yl		(99%)	D	608
137			116	D	635
138	Cl		28*	C	565
139	Cl		59	C	579
140	Cl		155	C	579
141	Cl		87*	C	566
142			24*	C	628
143			35*	C	658
144	Cl		14	C	648
145	Cyclopropyl		71	C	654
146	Thiophen-2-yl		85	C	696
147	Cl		81	C	565
148	Cl		75	C	583

Footnotes to Tables 2-4:

* Single enantiomer

** (% inhibition of calpain I at 10,000 nM)

149	Cl	CH	n-Bu		23	B	MH+576
150	Cl	CH			(78%)	B	562
151	Cl	CH	n-Bu		217	B	599
152	Cl	CH	n-Pr		325	B	585
153	Ph	N	Bn		(95%)	C	608
154		Ph N	Bn		83	C	691
155	Cl	CH	n-Bu :		31	B	614
156	Cl	CH	n-Pr		43	B	600
157	Cl	CH			61	B	603
158	Cl	CH			(94%)	B	519

** (% inhibition of calpain I at 10,000 nM)

100	360	36
96	375	66
103	435	380
104	330	1160
84	348	14
86	1030	475

Claims (23)

1. A compound having the formula I:wherein:

   X is CH, N, or CQ¹, with the proviso that when X is CQ¹, at least one of Q or Q¹ is H;

   R is selected from the group consisting of H, alkyl having from one to 6 carbons, arylalkyl having from 7 to 15 carbons, cyclo(heteroalkyl) in which the ring contains from 5 to 14 ring atoms, heteroarylC_1-6alkyl in which the heteroaryl ring contains from 5 to 14 ring atoms, C_1-10alkohyC_1-10alklyl, a side chain of a naturally occurring amino acid in the R or S configuration, and (CH₂)nNH-L, said alkyl, cycloalkyl, arylalkyl, heteroalkyl, and heteroarylalkyl groups being optionally substituted with one or more J groups;

   L is selected from the group consisting of alkoxycarbonyl having from 2 to 7 carbons, arylalkoxycarbonyl in which the arylalkoxy group contains 7 to 15 carbons, S(=O)₂R², and N-nitroimino;

   R² is selected from the group consisting of C_1-6alkyl, and aryl having from 6 to 14 carbons;

   R¹ is selected from the group consisting of H, halogen, cyano, nitro, sulfonic acid, hydroxyl, C_1-10alkyl, C_1-10alkoxy, hydroxymethyl, C_1-10alkoxymethyl, arylalkyl having from 7 to 15 carbons, carboxyl, C_1-1alkoxycarbonyl, C_1-10alkylcarbonyloxy, haloC_1-10alkyl, N(RR³), and C_1-10alkylcarbonyl;

   R³ is the same as R;

   Q is selected from the group consisting of H, C_1-6alkyl, C_3-10cycloalkyl, hydroxyl, C_1-10alkoxy, halogen, arylalkyl having from 7 to 15 carbons, arylalkenyl having from 8 to 16 carbons, arylalkynyl having from 8 to 16 carbons, aryl having from 6 to 14 carbons, heteroaryl having from 5 to 14 ring atoms, cyclo(heteroalkyl) having from 5 to 14 ring atoms, cycloalkyl having from 3 to 10 carbons. S-R, S(=O)R, S(=O)₂R, N(RR³), and NHS(7=O)₂R, said arylalkyl, arylalkenyl, arylalkynyl, aryl, heteroaryl, and heteroalkyl groups being optionally substituted with one or more J groups;

   Q¹ is the same as Q;

   Z is COCONH-R⁷;

   R⁷ is selected from the group consisting of K, -A-N(R⁸)-G, -O-A-N(R⁸)-G, -A-SO₂N(R⁸)(R⁹), and-O-A-SO₂N(R⁸)(R⁹);

   K is selected from the group consisting of C_1-10alkyl, alkenyl selected from ethenyl and propenyl, alkynyl selected from ethynyl and propynyl, C_3-10cycloalkyl, cyclo(heteroalkyl) having from 5 to 14 ring atoms, heteroaryl having from 5 to 14 ring atoms, C_6-14aryl, C_7-15arylalkyl, heterocycloalkyl having a ring having from 5 to 14 ring atoms attached through a C_1-6alkyl, C_1-10alkoxy, C_1-10alkoxyC_1-10alkyl, C_7-18arylalkyloxy, and N(RR³), said K groups being optionally substituted with one or more J groups;

   A is C_1-6alkylene optionally substituted with one or more J groups;

   R⁸ is selected from the group consisting of H and C_1-6alkyl;

   R⁹ is selected from the group consisting of H, C_1-10alkyl, C_6-14aryl, and heterocyclyl having from 5 to 14 ring atoms, said alkyl, aryl, and heterocyclyl groups being optionally substituted with one or more J groups;

   G is selected from the group consisting of C(=O)C_6-14aryl, C(=O)heteroaryl wherein the heteroaryl has from 5 to 14 ring atoms, C(=O)cyclo(hateroalkyl) wherein the cyclo(heteroalkyl) has from 5 to 14 ring atoms, C_1-10alkanoyl, C(=S)NH(C_6-14aryl), C(=O)NH(C_6-14aryl) C(=O)NH(C_3-10Cycloalkyl), CO₂-(C_6-14aryl), C(=O)C_1-10alkyl, CO₂(C_1-10alkyl), CO₂(C_7-15arylalkyl), C_1-10alkylsulfonyl, C_1-10alkenylsulfonyl, C_6-14arylsulfonyl, heteroarylsulfonyl wherein said heteroaryl has from 5 to 14 ring atoms, a side chain of a naturally occurring amino acid in the R or S configuration, a protecting group selected from the group consisting of benzyloxycarbonyl, toluenesulfonyl, t-butoxycarbonyl, methyl ester and benzyl ether groups, and SO₂(RR³), said G groups being optionally substituted with one or more J groups;

   J is selected from the group consisting of H, halogen, cyano, nitro, hydroxyl, C_1-10alkyl, C_1-10alkoxy, C_6-14aryl, C_7-15arylalkyl, C_1-10alkoxycarbonyl, C_1-10alkylcarbonyloxy, C_1-10alkenylcarbonyloxy, haloC_1-10alkyl, aminoC_1-10alkyl, haloC_1-10alkoxy, SO₂NRR³), SO₂NH(C_6-14aryl), SO₂NH(heteroaryl) wherein said heteroaryl has from 5 to 14 ring atoms, NHC(=O)NH(C_6-14aryl), NH(C=O)NH(heteroaryl) wherein said heteroaryl has from 5 to 14 ring atoms, NHSO₂(C_6-14aryl), NHC(=O)C_1-10alkyl, NHC(=O)C_6-14aryl, NHC(=O)heteroaryl wherein said heteroaryl has from 5 to 14 ring atoms, N(RR³), and NH=C(NH₂)₂, where R and R³ are not further substituted with J;

   n is an integer from 2 to 6;

   wherein alkyl includes straight-chain, branched and cyclic hydrocarbon groups; or a pharmaceutically acceptable salt thereof;

   with the proviso that when R⁷ is K, then Q is selected from the group consisting of optionally substituted arylalkenyl and optionally substituted arylalkynyl; and

   with the further proviso that when K is alkyl, then X is not CH when Q is optionally substituted arylalkynyl
2. The compound of claim 1 wherein X is CH.
3. The compound of claim 1 wherein R is selected from the group consisting of allcyl having from 2 to 4 carbons, and arylalkyl having from 7 to 15 carbons.
4. The compound of claim 3 wherein R is benzyl.
5. The compound of claim 1 wherein R¹ is H or C_1-10alkoxy.
6. The compound of claim 5 wherein R¹ is H.
7. The compound of claim 1 wherein Q is selected from the group consisting of arylalkynyl having from 8 to 16 carbons, aryl having from 6 to 14 carbons and halo.
8. The compound of claim 7 wherein Q is phenylC_2-10alkynyl.
9. The compound of claim 1 wherein A is selected from the group consisting of (CH₂)_n wherein n is 2 or 3, and (CH₂)_vCH₂-J where v is an integer from 1 to 6.
10. The compound of claim 9 wherein A is (CH₂)_vCH₂-J, where v is 2 or 3.
11. The compound of claim 1 wherein K is selected from the group consisting of C_1-10alkyl, hydroxyC_1-10alkyl, haloC_1-10alkyl, alkynyl selected from ethynyl and propynyl, heterocycloalkyl having a ring having from 5 to 14 ring atoms attached through a C_1-6alkyl, C_7-15arylalkyl, and cyclo(heteroalkyl) having from 5 to 14 ring atoms.
12. The compound of claim 1 wherein G is selected from the group consisting of substituted or unsubstituted C(=O)C_6-14aryl, C(=O)heteroaryl wherein the heteroaryl has from 5 to 14 ring atoms, C_6-14arylsulfonyl, and heteroarylsulfonyl wherein said heteroaryl has from 5 to 14 ring atoms.
13. The compound of claim 12 wherein G is selected from the group consisting of unsubstituted C_6-14arylsulfonyl, substituted C_6-14arylsulfonyl, unsubstituted and heteroarylsulfonyl wherein said heteroaryl has from 5 to 14 ring atoms and substituted and heteroarylsulfonyl wherein said heteroaryl has from 5 to 14 ring atoms.
14. The compound of claim 1 wherein X is CH, Z is COCONH-K, R₁ is H, and R is selected from the group consisting of alkyl having from 2 to 4 carbons and arylalkyl having from 7 to 15 carbons.
15. The compound of claim 14 wherein R is benzyl.
16. The compound of claim 1 wherein X is CH, Z is COCONH-K, R₁ is H, R is selected from the group consisting of alkyl having from 2 to 4 carbons and arylalkyl having from 7 to 15 carbons, and Q is arylalkynyl having from 8 to 16 carbons.
17. The compound of claim 16 wherein R is benzyl.
18. The compound of claim 16 wherein Q is phenylethynyl.
19. The compound of claim 1 wherein Q is phenylethynyl; X is =CH-; R¹ is H; R is benzyl; and Z is COCONH-R⁷.
20. A compound of formula Ia, Ib or Ic:selected from the following Tables:1phenylethynylbenzylIa2phenylethynylbenzylIa3phenylethynylbenzylIa4phenylethynylbenzylIa5phenylethynylbenzylIa6phenylethynylbenzylIa7phenylethynylbenzylIa8phenylethynylbenzylIa9phenylethynylbenzylIa10phenylethynylbenzylIa11phenylethynylbenzylIa12phenylethynylbenzylIa13phenylethynylbenzylIa14phenylethynylbenzylIa15adamantylphenylethynylbenzylIa16cyclopropanephenylethynylbenzylIa17(1-benzyl)piperidin4-ylphenylethynylbenzylIa18(2-methylcyclohexyl)phenylethynylbenzylIa19(4-methylcyclohexyl)phenylethynylbenzylIa20(3-methylcyclohexyl)phenylethynylbenzylIa21phenylethynylbenzylIa22ε-caprolactamphenylethynylbenzylIa23phenylethynylbenzylIa24phenylethynylbenzylIa25phenylethynylbenzylIa26phenylethynylbenzylIa27phenylethynylbenzylIa28cyclohexan-2-olphenylethynylbenzylIa29[trans]cyclohexan4-olphenylethynylbenzylIa30cyclohexylphenylethynylbenzylIa31phenylethynylbenzylIa32tetrahydronaphth-1-ylphenylethynylbenzylIa33piperon-5-ylphenylethynylbenzylIa34phenylethynylbenzylIa35benzodioxan-6-ylphenylethynylbenzylIa36phenylethynylbenzylIa37phenylethynylbenzylIa38phenylethynylbenzylIa39phenylethynylbenzylIa40phenylethynylbenzylIa41phenylethynylbenzylIa42phenylethynylbenzylIa43phenylethynylbenzylIa44phenylethynylbenzylIa45phenylethynylbenzylIa46phenylethynylbenzylIa47phenylethynylbenzyl-Ia48phenylethynylbenzylIa49phenylethynylbenzylIa50phenylethynylbenzylIa51phenylethynylbenzylIa52phenylethynylbenzylIa53phenylethynylbenzylIa54phenylethynylbenzylIa55indan-2-ylphenylethynylbenzylIa56phenylethynylbenzylIa57phenylethynylbenzylIa58phenylethynylbenzylIa59phenylethynylbenzylIa60phenylethynylbenzylIa61phenylethynylbenzylIa62phenylethynylbenzylIa63phenylethynylbenzylIa64phenylethynylbenzylIa65phenylethynylbenzylIa66phenylethynylbenzylIa67phenylethynylbenzylIa68phenylethynylbenzylIa69phenylethynylbenzylIa70phenylethynylbenzylIa71phenylethynylbenzylIa72phenylethynylbenzylIa73phenylethynylbenzylIa74phenylethynylbenzylIa75phenylethynylbenzylIa76phenylethynylbenzylIa77phenylethynylbenzylIa78phenylethynylbenzylIa79phenylethynylbenzylIa80phenylethynylbenzylIa81phenylethynylbenzylIa82phenylethynylbenzylla83phenylethynylbenzylla84phenylethynylbenzylla85phenylethynylbenzylIa86phenylethynylbenzylla87phenylethynylbenzylla88phenylethynylbenzylIa89phenylethynylbenzylIa90phenylethynylbenzylIa91phenylethynylbenzylIa92phenylethynylbenzylIa93phenylethynylbenzylIa94phenylethynylbenzylIa95phenylethynylbenzylIa96phenylethynylbenzylIa97phenylethynylbenzylIa98phenylethynylbenzylIa99phenylethynylbenzylIa100phenylethynylbenzylIa101phenylethynylbenzylIa102phenylethynylbenzylIa103phenylethynylbenzylIa104OBnphenylethynylbenzylIa105phenylethynylbenzylIa106phenylethynylbenzylIa107phenylethynylbenzylIa108phenylethynylbenzylIa109phenylethynylbenzylIa110phenylethynylbenzylIa111phenylethynylbenzylIa112iBuphenylethynylIa113iBuphenylethynylIa114iBuphenylethynylIa115iBuphenylethynylIa116iBuphenylethynylIa117iBuphenylethynylIa118iBuphenylethynylIa119iBuphenylethynylIa120EtphenylethynylIa121EtphenylethynylIa122EtphenylethynylIa123EtphenylethynylIa124EtphenylethynylIa125EtphenylethynylIa126BuphenylethynylIa127BuphenylethynylIa128PrphenylethynylIa129PrphenylethynylIa130PhCH=CHBubenzylIa131HbenzylIa132H(N-oxide)benzylIa133HObenzylIa134benzylIa135Piperidin-1-ylbenzylIa136Pyridin-3-ylbenzylIa137benzylIa138ClbenzylIa139ClbenzylIa140ClbenzylIa141ClbenzylIa142benzylIa143benzylIc144ClbenzylIa145Cyclo-propylbenzylIa146Thiophen-2-ylbenzylIa147ClbenzylIa148ClbenzylIa149Cln-BuIa150Cln-PrIa151Cln-BuIa152Cln-PrIa153PhBnIb154PhBnIb155Cln-BuIa156Cln-PrIa157ClIa158ClIaor a pharmaceutically acceptable salt thereof.
21. A composition for inhibiting a serine protease or a cysteine protease comprising a compound of claim 1 and a pharmaceutically acceptable carrier.
22. The compound of claim 1 for use in therapy.
23. The use of a compound of claim 1 for the manufacture of a medicament for the treatment of a disease associated with abnormal and/or aberrant activity of cysteine proteases and/or serine proteases.